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STUDIES ON THE CHARACTERISTICS OF PHYTOMASS-BASED VERMICOMPOSTS AND THEIR INFLUENCE ON THE PLANT GROWTH

Thesis submitted

for the award of the degree of

DOCTOR OF PHILOSOPHY

in

Environmental Technology

by

M. Karthikeyan, MSc., MPhil

Under the supervision of

Dr. S. Gajalakshmi, MSc., MPhil., Ph.D

Assistant Professor

And Co-guidance of

Prof. S. A. Abbasi, PhD, DSc, FNASc, FIIChE, FIE, PE

Senior Professor and Head

Centre for Pollution Control and Environmental Engineering Pondicherry (Central) University Puducherry - 605014, India

CERTIFICATE

This is to certify that Mr. M. Karthikeyan has carried out the work embodied in his thesis

entitled ‘Studies on the characteristics of phytomass-based vermicomposts and their

influence on the plant growth’ being submitted to Pondicherry University for the award of

the degree of Doctor of Philosophy in Environmental Technology. He has complied with all

the relevant academic and administrative regulations, and the thesis embodies a bonafide

record of the work done by him under our guidance. The work is original and has not been

submitted for the award of any certificate, diploma, or degree, of this or any other university.

Dr. S. Gajalakshmi Prof. S. A. Abbasi

Supervisor Co-guide

DECLARATION

I hereby declare that the thesis entitled “Studies on the characteristics of phytomass-based

vermicomposts and their influence on the plant growth” submitted to Pondicherry

University for the award of the degree of Doctor of Philosophy is a record of original work

done by me under the guidance of Dr. S. Gajalakshmi, Assistant Professor, and co-guidance

of Prof. S. A. Abbasi, Senior Professor and Head, Centre for Pollution Control and

Environmental Engineering, Pondicherry University, and that it has not formed the basis for

the award of any other degree, diploma, certificate or any other title by any university or

institution before.

Date:

Place: Puducherry (M. Karthikeyan)

It may be doubted whether there are many other animals which have played so important a part in the history of the world, as have these lowly organized creatures.

Charles. R. Darwin (1881)

To My Parents

To My Wife Vimala and

Daughter Aaradhana

Acknowledgements

“Kindness shown by those who weigh not what the return may be:

When you ponder right its merit, 'Tis vaster than the sea” - Thirukural

After years and years of receiving support and help from family, teachers and friends, finally

time has come to thank these wonderful people for their unselfish contribution.

First of all I thank my thesis supervisor Dr. S. Gajalakshmi for her care and encouragement

especially during the difficult times, and for having confidence in me and my work. “Mam,

without your support, I would have not achieved this stage. I have no words to express my

gratefulness, so I salute you, mam”.

I am grateful to Prof. S.A. Abbasi. His wide knowledge, „decades and decades of experience‟

in this field and his logical way of approaching problems have been of great value for me. His

genuine and open-minded curiosity to learn new things has inspired me all over these years.

I would also like to acknowledge my doctoral committee member, Dr. D. Senthilnathan for

his stimulating decisions and valuable comments on my work. Other than my work, he wanted

me to submit thesis early. I never had seen such a kind-hearted teacher in my life, “Thank you

sir”.

I thank Dr. Tasneem Abbasi and Er. S.Sudalai for their timely help, and they never denied or

delayed any assistance whenever I have requested them. I also sincerely thank my RGNF

committee member Dr. E.V. Ramasamy for his valuable input in my research work.

I own my sincere gratitude to all my Teachers and Professors who have shaped me to what I

am. Especially my Maths teacher Mr. S. Jayaraj, post-graduate teacher Dr. P. Suresh and

Dr. Christabelle E.G. Fernandes, without them I would have not been able to reach this

stage. I have been so blessed to get these wonderful people in my life.

பயன்தூக்கார் செய்த உதவி நயன்தூக்கின்

நன்மை கடலின் சபரிது

I am indebted to Mr. Prabanandham for his help and care all along, “Anna, you presence

always made me laugh”. Special thanks to Mr. Premaravind for helping me in fixing both lab

and personal needs.

I am also grateful to Mr. Karunakaran, Mr. Balamurugan and Mr. Vetrivel, Department of

Earth Science, for helping me in thin-sectioning and ICP analysis. I must thank Er. Ramasamy

and Mrs. S. Elisa Fathima, Central Instrumentation Facility, Mr. Anbalagan, Sophisticated

Analytical Instrument Facility, IITM, Chennai, and Mr. Adarsh, Sophisticated Test and

Instrumentation Centre, Cochin University, Cochin for technical support.

Many thanks to mera dost Ajay Harit for his contribution in all my experimental work. Also I

thank my friends Sudakaran (Ecology), Babuji (Management studies), Vinoth (Physical

Education) and Gnanavel (Chemistry) for being there for my needs and I really had a wonderful

time with them during these „Ph.D years‟. Heartfelt thanks to my beloved friends Baskaran

(Marine Biology), Anandha Dhanaselvan (CPEE), Purushothaman (Philosophy) and

Srinivasan (Alagappa University) for their help and support.

I cannot forget the help of M.Phil scholar Hariharan, who have worked with me day and night

to set the plant growth studies. I thank my friend Nayeem Shah (alias Peter) for his fun full

company, and also I thank Antony Godson, Nasser Hussain and Mohammad Ashraf for the

same. Many people in CPEE have decorated my memories of „Ph.D years‟, for this I should

thank Dr. R. Sanjeevi, Dr. J. Anuradha, Dr. G. Ponni, Dr. Manoj Makhiija, Mrs. Gurjeet

Kaur, Mrs. P. Poornima, Mr. T. Ganesh, Mr. T.V. Ananthaaraj and so many… :-)

No words to either thank or praise my Parents and Lord Almighty who gave me every possible

support throughout and holding my hand each step of the way. I thank my wife for her

patience, love and support. I cordially thank my sister. ‘Indu! you are my strength!’ I thank

GOD for having blessed me with such a loving sister.

Last but not least, I would like to thank my lovely worms for its great job! “Buddies! I will not

disturb you anymore! Enjoy!”

M.Karthikeyan

i

Contents

List of Tables ……………………………………………………………………………………... vii-x

List of Figures …………………………………………………………………………………….. xi-xii

Chapter 1. Introduction 01

Chapter 2. Vermicompost characteristics and its effect on soil and plant growth – state-of-

the-art

Abstract………………………………………………………………………………………... 05

1. Introduction ………………………………………………………………………………... 06

2. Types of vermicast and its composition …………………………………………………… 06

2.1. Types of vermicast ………………………………………………………………… 06

2.1.1. Based on place of deposition: surface and subterranean castings ………... 06

2.1.2. Based on shape of castings ……………………………………………….. 06

2.1.3. Based on size of the castings ……………………………………………... 10

2.2. Physical properties of vermicast …………………………………………………... 11

2.2.1. Color and odor ……………………………………………………………. 11

2.2.2. Bulk density ………………………………………………………………. 11

2.2.3. Pore space ………………………………………………………………… 13

2.2.4. Water-related properties ………………………………………………….. 14

2.3. Chemical properties of vermicast …………………………………………………. 15

2.3.1. Organic carbon ……………………………………………………………. 15

2.3.2. Macronutrients (primary) ………………………………………………..... 20

2.3.3. Macronutrients (secondary) ………………………………………………. 23

2.3.4. Micronutrients …………………………………………………………….. 25

3. Effect of ageing on the properties of vermicast …………………………………………… 29

4. Effect of vermicast on plant growth and soil ………………………………………………. 31

5. Conclusion …………………………………………………………………………………. 32

References ……………………………………………………………………………………. 33

Chapter 3. Ingestion of sand and soil by phytophagous earthworm Eudrilus eugeniae: a

finding of relevance to earthworm ecology as well as vermitechnology

Abstract ……………………………………………………………………………………….. 53

1. Introduction ………………………………………………………………………………... 53

2. Materials and methods ……………………………………………………………………... 54

3. Results and discussion ……………………………………………………………………... 56

3.1. Vermicast output ………………………………………………………………….. 56

3.2. Mortality …………………………………………………………………………... 58

ii

3.3. Average zoomass change per animal ……………………………………………... 58

3.4. Surface area of vermicast …………………………………………………...…….. 58

3.5. Assimilation of sand and soil particles in castings ……………………………… 59

4. Conclusions ………………………………………………………………………………... 61

References ……………………………………………………………………………………. 61

Chapter 4. Feeding behaviour of phytophagous earthworm Eudrilus eugeniae in high-

substrate column vermireactors

Abstract ……………………………………………………………………………………….. 65

1. Introduction ………………………………………………………………………...……… 65

2. Materials and methods …………………………………………………………...………… 66

3. Results and discussion …………………………………………………………………… 67

4. Conclusions ………………………………………………………………………………... 69

References ……………………………………………………………………………………. 69

Chapter 5. Effect of sand and soil ingestion by phytophagous earthworm Eudrilus eugeniae

on the physical and chemical properties of vermicast

Abstract ……………………………………………………………………………………….. 71

1. Introduction ………………………………………………………………………………... 71

2. Materials and methods ……………………………………………………...………….…... 72

2.1. Experimental design ………………………………………………………………. 72

2.2. Vermireactors operation …………………………………………………………... 72

2.3. Analytical methods ……………………………………………...………………… 72

2.4. Assessment of soil/sand content in the vermicast ………………………………… 73

2.5. Data analysis ……………………………………………………...……………….. 73


3.1. Assimilation of sand/soil in the vermicast ………………………………………… 73

3.2. Vermicast output ………………………………………………………………….. 75

3.3. Growth and survival of earthworms ………………………………………………. 75

3.4. Physical and chemical properties of vermicast …………………………………… 76

4. Conclusions ………………………………………………………………..………………. 77

References ……………………………………………………………………………………. 78

Chapter 6. Effect of vermicast generated from an allelopathic weed lantana (Lantana

camara) on seed germination, plant growth, and yield of cluster bean (Cyamopsis

tetragonoloba)

Abstract ……………………………………………………………………………………….. 81

1. Introduction ……………………………………………………………………………… 81


2.1. Germination, plant growth and yield characteristics ……………………………… 83

2.3. Analytical methods ………………………………………...……………………… 84

iii

2.4. Statistical analysis …………………………………………………….………… 85


3.1. Seed germination ………………………………………………………………… 85

3.2. Plant growth …………………………………………..…………………………… 86

3.3. Photosynthetic pigments ……………………………..…………………………… 88

3.4. Flowering ………………………………………………………………………….. 88

3.5. Disease incidence, plant death and stunted growth ……………………………….. 88

3.5. Fruit yield ………………………………………….……………………………. 89

3.6. The present study in the context of the state-of-the-art ………...…………………. 90

4. Conclusions ………………………………………………………………………………. 91

References ……………………………………………………………………………………. 91

Supplementary material ………………………………………………………………............. 96

Chapter 7. Effect of vermicast generated from a pernicious weed ipomoea (Ipomoea

carnea) on seed germination, plant growth, and yield of cluster bean (Cyamopsis

tetragonoloba)

Abstract ……………………………………………………………………………………….. 101

1. Introduction ……………………………………………...………………………………… 101

2. Materials and methods ……………………………………………………………………. 102


2.3. Analytical methods …………………………………...…………………………… 104

2.4. Statistical analysis ………………………………………………………………… 104

3. Results and discussion …………………………………………………………………….. 104

3.1. Seed germination ………..………………………………………………………… 104

3.2. Plant growth …………….…………………………………………………………. 105

3.3. Photosynthetic pigments …...……………………………………………………… 107

3.4. Effect on flowering …………...…………………………………………………… 107


3.6. Effect on yield …………………………………………………………………….. 108

4. Conclusions ………………………………………..………………………………………. 109

References ……………………………………………………………………………………. 109

Supplementary material ………………………………………………………………………. 112

Chapter 8. Comparative efficacy of vermicomposted paper waste and inorganic fertilizer

on seed germination, plant growth and fruition of cluster bean (Cyamopsis tetragonoloba)

Abstract ……………………………………………………………………………………….. 117

1. Introduction ………………………………………………………………………………... 117

2. Materials and methods …………………………………………………………………… 118

2.1. Study area …………………………………………………………………………. 118

2.2. Treatments ………………………………………………………………………… 118


iv

2.3. Analytical methods ………………………………………………………………... 120

2.4. Statistical analysis ………………………………………………………………… 120


3.1. Seed germination ………………………………………………………………... 120

3.2. Plant growth ……………………………………………………………………... 121

3.3. Photosynthetic pigments ………………………………………………………… 123


3.5. Effect on flowering ……………………………………………………………… 123

3.6. Effect on yield …………………………………………………………………….. 123

4. Conclusions ………………………………………………………………………………... 124

References ……………………………………………………………………………………. 124

Supplementary material ………………………………………………………………………. 127

Chapter 9. Effect of vermicast generated from allelopathic weeds and paper waste on

physical and chemical properties of potting soil growing cluster bean (Cyamopsis

tetragonoloba)

Abstract ………………………………………………………………………………………. 131

1. Introduction ……………………………………………………………………………….. 131

2. Materials and methods …………………………………………………………………….. 132

3. Results and discussion ………………………………………………………………….. 134

3.1. Physical properties ………………………………………………………………. 134

3.2. Chemical properties …………………………………………………………….. 142

4. Conclusions ………………………………………………………………………………. 154

References …………………………………………………………………………………… 155

Chapter 10. Effect of storage on some physical and chemical characteristics of vermicast:

A preliminary study

Abstract ……………………………………………………………………………………… 159

1. Introduction ……………………………………………………………………………… 159



4. Conclusions ………………………………………………………………………………... 164

References ……………………………………………………………………………………. 164

Chapter 11. Effect of storage on the properties of vermicompost generated from paper

waste – with focus on pre-drying and extent of sealing

Abstract ……………………………………………………………………………………….. 169

1. Introduction ………………………………………………………………………………... 169


2.1. Types of storage ………………………………………………………………… 170

2.2. Analysis …………………………………………………………………………… 171

v

2.3. Processing of data …………………………………………………………………. 171


3.1. Physical properties ………………………………………………………………… 171

3.2. Chemical properties ……………………………………………………………….. 173

3.3. Biochemical properties ……………………………………………………………. 177

4. Conclusions ……………………………………………………………………………… 180

References …………………………………………………………………………………… 180

Chapter 12. Effect of pre-drying and extent of sealing on the properties of vermicast

generated from the neem leaves during storage

Abstract ……………………………………………………………………………………….. 185

1. Introduction ………………………………………………………………………………. 185


2.1. Experimental set up ……………………………………………………………….. 186


2.3. Data analysis ………………………………………………………………………. 186





4. Conclusions ………………………………………………………………………………... 198

References ……………………………………………………………………………………. 198

Chapter 13. Effect of pre-drying and extent of sealing on the properties of vermicomposted

cow dung during storage

Abstract ……………………………………………………………………………………….. 203

1. Introduction ………………………………………………………………………………... 203

2. Materials and methods …………………………………………………………………… 203

2.1. Experimental design ………………………………………………………………. 203


2.3. Data analysis ………………………………………………………………………. 204





4. Conclusions ………………………………………………………………………………... 214

References …………………………………………………………………………………..... 215

Chapter 14. Summary and conclusion 219

Appendix – Standardization of analytical methods 223

vii

List of Tables Chapter 2

1. Shape and size of the castings produced by different earthworm species ……………………. 08

2. Some of the chemical properties of castings produced by different earthworm species from

different organic wastes. ……………………………………………………………………… 17

3. Some of the micronutrient content of castings produced by the different earthworm species

from the different organic wastes…………………………………………………………… 27

Chapter 3

1. Major and trace element composition of plant leaves, cow dung, soil and sand used in this

study …………………………………………………………………………………………. 55

2. Repeated analysis of variants and ANOVA table of F-values and the effects of substrate,

bedding and worm density on vermicast output, average zoomass changes, mortality, sand

and soil entrapped in castings, castings surface area and sand grains covered area of

castings………………………………………………………………………………………... 57

Chapter 5

1. The physical properties of vermicast generated from the reactors without sand/soil + 250

and 500 g substrate and reactors consisting sand/soil + 250 and 500 g substrate…...……...… 76

2. The chemical properties of vermicast generated from the reactors without sand/soil + 250

and 500 g substrate and reactors consisting sand/soil + 250 and 500 g substrate………..…… 77

Chapter 6

1. Chemical and physical properties of vermicast and soil used in the study ………………… 84

2. Amount of inorganic fertilizer applied equivalent to vermicast treatment……………………. 85

3. Germination value and germination percentage of the seeds of cluster bean as influenced by

lantana vermicast and inorganic fertilizers…………………………………………………… 86

4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth stunned

growth/death in cluster bean plants as impacted by lantana vermicast or inorganic

fertilizers……………………………………………………………………………………… 87

5. Harvest index and yield attributes of plants as impacted by lantana vermicast and equivalent

inorganic fertilizers……………………………………….…………………………………… 89

Chapter 7


2. Amount of inorganic fertilizer applied equivalent to vermicast treatment …………………… 104


ipomoea vermicast and inorganic fertilizers………………………………….……………… 105


growth/death in cluster bean plants as impacted by ipomoea vermicast or inorganic

fertilizers…………………………………………………………………………………….. 106

5. Harvest index and yield attributes of plants as impacted by ipomoea vermicast and

equivalent inorganic fertilizers……………………………………………………………….. 108

viii

Chapter 8

1. Chemical and physical properties of vermicast and soil used in the study …………………... 119

2. Amount of inorganic fertilizer applied equivalent to vermicast treatment …………………… 120


paper waste vermicast and inorganic fertilizers……………….…………….……………… 121


growth/death in cluster bean plants as impacted by paper waste vermicast or inorganic

fertilizers………………………...…………………………………………………………….. 122

Chapter 9


2. Changes in the bulk density of potting soil amended with vermicast from lantana, ipomoea

and paper waste, or equivalent inorganic fertilizers, at different periods of time ……………. 135

3. Changes in the particle density of potting soil amended with vermicast from lantana,

ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time … 136

4. Changes in the total porosity of potting soil amended with vermicast from lantana, ipomoea

and paper waste, or equivalent inorganic fertilizers, at different periods of time…………….. 137

5. Changes in the water filled pore space of potting soil amended with vermicast from lantana,

ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time… 138

6. Changes in the water holding capacity of potting soil amended with vermicast from lantana,

ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time…... 139

7. Changes in the electrical conductivity of potting soil amended with vermicast from lantana,

ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time…... 140

8. Changes in the pH of potting soil amended with vermicast from lantana, ipomoea and paper

waste, or equivalent inorganic fertilizers, at different periods of time ……………………….. 141

9. Changes in the organic carbon of potting soil amended with vermicast from lantana,

ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time… 144

10. Changes in the total nitrogen of potting soil amended with vermicast from lantana, ipomoea

and paper waste, or equivalent inorganic fertilizers, at different periods of time ……………. 145

11. Changes in the ammonia of potting soil amended with vermicast from lantana, ipomoea and

paper waste, or equivalent inorganic fertilizers, at different periods of time ………………… 146

12. Changes in the nitrate of potting soil amended with vermicast from lantana, ipomoea and

paper waste, or equivalent inorganic fertilizers, at different periods of time ………………… 147

13. Changes in the exchangeable phosphorus of potting soil amended with vermicast from

lantana, ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of

time …………………………………………………………………………………………… 148

14. Changes in the exchangeable potassium of potting soil amended with vermicast from

lantana, ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of

time …………………………………………………………………………………………… 149

15. Changes in the exchangeable calcium of potting soil amended with vermicast from lantana,

ipomoea and paper waste, or equivalent inorganic fertilizers, at different periods of time….. 150

16. Changes in the trace elements of potting soil amended with vermicast from lantana or

equivalent inorganic fertilizers, at different periods of time………………………………… 151

17. Changes in the trace elements of potting soil amended with vermicast from ipomoea or

equivalent inorganic fertilizers, at different periods of time …………………………………. 152

18. Changes in the trace elements of potting soil amended with vermicast from paper waste or

equivalent inorganic fertilizers, at different periods of time …………………………………. 153

ix

Chapter 10

1. Physical characteristics of castings stored for different periods and the calculated F-values

using one-way ANOVA ……………………………………………………………………… 162

2. Total nitrogen and organic carbon of castings stored for different periods and the calculated

F-values using one-way ANOVA ……………………………………………………………. 162

3. Major elements present in the casting stored for different periods …………………………... 163

Chapter 11

1. F values of repeated measures analysis of variance on the effect of extend of sealing and

pre-treatment on physical properties, EC and pH of vermicast during the storage…………… 174


pre-treatment on chemical properties of vermicast during the storage……………………….. 174


pre-treatment on biochemical properties of vermicast during the storage……………………. 174

Chapter 12





3. F values of repeated measures analysis of variance on effect of extend of sealing and pre-

treatment on biochemical properties of vermicast during the storage………………………… 189

Chapter 13






pre-treatment on biochemical properties of vermicast during the storage.…………………… 209

xi

List of Figures Chapter 3

1. Vermicast output, g worm-1

day-1

recorded in reactors with different treatments ……………. 56

2. Percentage of sand and soil particle entrapped in the castings measured by gravimetric

method and percentage of sand particle covered area in the castings measured by thin-

sectioning method …………………………………………………………………………….. 59

3. The thin sectioned vermicast from neem, cow dung and ipomoea based reactors with or

without soil + sand bedding……………….…..………………………………………………. 60

4. Regression analysis between the sand and soil entrapped in the castings and (a) the average

zoomass changes and (b) mortality rate …………………………………………………….... 60

Chapter 4

1. Percentage of sand and soil particle entrapped in the castings of different treatments ………. 67

2. Vermicast output, grams worm-1

day-1

recorded in reactors with different treatment ………... 68

Chapter 5

1. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil

with 250 g and 500 g substrate in the first trial ………………………………………………. 73

2. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil

with 250 g and 500 g substrate in the second trial …………………………………………... 74

3. Vermicast output, grams worm -1 day-1 recorded in reactors without sand/soil + 250 and

500 g substrate and reactors consisting sand/soil + 250 and 500 g substrate ………………… 75

Chapter 11

1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding

capacity (d) of un-dried and pre-dried castings stored in airtight sealed bags and un-dried

and pre-dried castings stored in partially sealed bags, at different periods of time ………….. 172

2. Changes in the total porosity (a), water-filled porosity (b), pH (c) and EC (d) of un-dried and

pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings stored in

partially sealed bags, at different periods of time …………………………………………….. 173

3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c),

ammonium nitrogen (d), nitrate nitrogen (e) and available phosphorus (f) content of un-dried

and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried castings

stored in partially sealed bags, at different periods of time …………………………………... 176

4. Changes in the extractable form of potassium (a), sulfur (b), calcium (c) and sodium (d)

content of un-dried and pre-dried castings stored in airtight sealed bags and un-dried and

pre-dried castings stored in partially sealed bags, at different periods of time ………………. 177

5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline

phosphatase (e) and arylsulphatase (f) enzymes activity of un-dried and pre-dried castings

stored in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed

bags, at different periods of time …………………………………………………………… 178

6. Changes in the microbial biomass carbon content of un-dried and pre-dried castings stored

in airtight sealed bags and un-dried and pre-dried castings stored in partially sealed bags, at

different periods of time ……………………………………………………………………… 179

xii

Chapter 12


capacity (d), of un-dried and pre-dried castings stored in airtight sealed bags and un-dried

and pre-dried castings stored in partially sealed bags, at different periods of time ………… 187

2. Changes in the total porosity (a), water-filled porosity (b), pH (c) and EC (d), of un-dried


stored in partially sealed bags, at different periods of time ………………………………… 188


ammonium nitrogen (d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and


partially sealed bags, at different periods of time …………………………………………. 191

4. Changes in the extractable form of potassium (a), sulfur (b), calcium (c) and sodium (d) of

un-dried and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried

castings stored in partially sealed bags, at different periods of time ……………………..... 192




bags, at different periods of time ……………………………………………………………... 195



different periods of time ……………………………………………………………………… 197

Chapter 13


capacity (d), of un-dried and pre-dried castings stored in airtight sealed bags and un-dried

and pre-dried castings stored in partially sealed bags, at different periods of time ………… 205

2. Changes in the total porosity (a), water-filled porosity (b), pH (c) and EC (d), of un-dried


stored in partially sealed bags, at different periods time …………………………………… 206


ammonium nitrogen (d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and


partially sealed bags, at different periods of time …………………………………………. 207

4. Changes in the extractable form of potassium (a), sulfur (b), calcium (c) and sodium (d) of

un-dried and pre-dried castings stored in airtight sealed bags and un-dried and pre-dried

castings stored in partially sealed bags, at different periods of time …………………………. 210




bags, at different periods of time …………………………………………………………… 211



different periods of time …………………………………………………………………….... 214

INTRODUCTION

Chapter

1

1

CChhaapptteerr 11

Introduction

Vermicomposting has become increasingly

popular across the world in recent decades

(Gajalakshmi and Abbasi, 2008; Edwards et al.,

2011). The ability of this process to convert various

types of biodegradable solid wastes into organic

fertilizer in a relatively low energy consuming and

pollution-generating manner has been the reason

behind this happening. Vermicompost is stabilized

organic waste by microbial bio-oxidation with

specific mediation of earthworms. It is reported to

improve seed germination, enhance seedling

growth and development, and increase plant

productivity even with a relatively small proportion

incorporated into growing media (Atiyeh et al.,

1998; Edwards et al., 2011). The extent of these

beneficial impacts of vermicompost on plants

depends on various factors. The nutrient content,

microbial activity, hydraulic properties and growth

regulators present in the vermicompost are crucially

important. All these beneficial properties of

vermicompost are subject to various changes with

substrate type, earthworm species and age of

castings. The dynamics of all these properties of

vermicast and its impact on plant growth and soil is

reviewed in Chapter 2.

The Chapters 3-5 comprise of the studies on

the feeding behavior of epigeic earthworms, which

are the species extensively used for

vermicomposting of various types of organic waste

(Abbasi and Ramasamy, 2001; Gajalakshmi et al.,

2001, 2002, 2005; Gajalakshmi and Abbasi,

2004a,b; Garg and Kaushik, 2005; Lim et al., 2012,

2014; Shak et al., 2014). The epigeics are

phytophagous earthworms which dwell at or very

near the surface of the soil horizon and feed upon

humus. The other two groups – anecics and

endogeics – make deep burrows, often live in

subsoil and due to their tendency to ingest soil they

are called as geophytophagous and geophagous

respectively. Many studies on anecics and

endogeics claim that ingestion of soil particles with

organic matter facilitates assimilation of nutrients

in earthworms gut probably by enhancing the

grinding action of the gizzard (Hendriksen, 1991;

Schulmann and Tiunov, 1999; Marhan and Scheu,

2005; Curry and Schmidt, 2007). But there are no

reports on the ingestion of sand particles by epigeic

earthworms. To fill this knowledge gap, attempt has

been taken to explore the influence of different

phytomass and worm density on ingestion of

soil/sand particles by epigeics and reported in

Chapters 3 and 4. The effect of mineral soil particle

ingestion on physico-chemical properties of

vermicast is studied and reported in Chapter 5.

In general, vermicast contains all the

essential macro and micronutrients for plant

growth, and it also improves the physical properties

of the soil, such as aeration, water holding capacity

and porosity of soil, all of which have direct impact

on the plant productivity (Edwards, 2004;

Gajalakshmi and Abbasi, 2008; Edward et al.,

2011). But it has to be ascertained whether the

vermicast generated from the invasive plants are

also beneficial to plants, as they are widely reported

to suppress the growth of other plant species.

Therefore, attempt has been made to assess the

effect of vermicompost generated from allelopathic

weeds Lantana camara and Ipomoea carnea on the

germination and growth of cluster bean (Cyamopsis

tetragonoloba) and reported in Chapters 6-8. The

response of this plant to vermicompost generated

from these weeds has been compared with

2

vermicompost from paper waste and equivalent

inorganic fertilizer treatments. The findings of these

studies revealed that the vermicompost derived

from different parent materials have different

physical, chemical and biological qualities, and

their impact on germination, growth and yield of

plant also varied considerably. To understand the

factors which attribute the differential impact of

vermicompost from different parent materials on

the growth and yields of plants, it is necessary to

recognize their impact on soil health. Hence, the

changes in the physical and chemical properties of

potting soil housing cluster bean (Cyamopsis

tetragonoloba) with the application of vermicast

generated from different organic wastes such as

paper waste, leaves of ipomoea (Ipomoea carnea),

and of lantana (Lantana camara) was studied and

reported in Chapter 9.

Although there are numerous reports on

vermicomposting and its application, there seems to

be an urgent need for the study of storage aspects of

vermicompost. There is a necessity to assess what

changes occurs in physico-chemical and biological

properties of vermicast during storage, so as to

work out the best strategy for packing and storing

the vermicompost which can enable retention of its

fertilizer value for long duration. In this regard,

attempts have been made towards formulating

packing guidelines for storing vermicast. The

findings of the study are briefed in the Chapters 10-

13. The summary of the findings of all the studies is

presented in Chapter 14. A brief account on the

standardization of analytical methods used in the

experiments reported in Chapters 3-13 is given in

Chapter 15.

References

Abbasi, S.A., Ramasamy, E.V., 2001. Solid waste

management with earthworms. Discovery

Publishing House, New Delhi. p.178.

Atiyeh, R.M., Subler, S., Edwards, C.A., 1998.

Growth of tomato plants in vermicomposted

hog manure. In proceedings of the 6th

International Symposium on Earthworm

Ecology Pedobiologia 43, 724–728.

Curry, J.P., Schmidt, O., 2007. The feeding ecology

of earthworms - a review. Pedobiologia 50,

463-477.

Edwards, C.A., Arancon, N.Q., Sherman, R., 2011.

Vermiculture technology: earthworms, organic

wastes and environmental management. CRC

Press, Boca Raton. pp. 103–164.

Gajalakshmi, S., 2002. Development of methods

for treatment and reuse of municipal and

agricultural solid wastes appropriate for

rural/suburban households. Ph.D thesis,

Pondicherry University, Puducherry. pp. 187.

Gajalakshmi, S., Abbasi, S.A., 2004a. Vermi-

conversion of paper waste by earthworm born

and grown in the waste-fed reactors compared

to the pioneers raised to adulthood on cow

dung feed. Bioresour. Technol. 94: 53–56.

Gajalakshmi, S., Abbasi, S.A., 2004b. Neem leaves

as a source of fertilizer-cum-pesticide

vermicompost. Biores. Technol. 92, 291-296.

Gajalakshmi, S., Abbasi, S.A., 2008. Solid waste

management by composting: state of the art.

Crit. Rev. Environ. Sci. Technol. 38, 311-400.

Gajalakshmi, S., Ramasamy, E.V., Abbasi, S.A.,

2001. Assessment of sustainable vermi-

conversion of water hyacinth at different

reactor efficiencies employing Eudrilus

eugeniae Kinberg. Bioresour. Technol. 80,

131-135.


2002. Vermicomposting of different forms of

water hyacinth by the earthworm Eudrilus

eugeniae, Kinberg. Bioresour. Technol.

82:165–169.


2005. Composting–vermicomposting of leaf

litter ensuing from the trees of mango

(Mangifera indica). Bioresour. Technol.

96:1057–1061.

Ganesh, P.S., 2007. Some application of bioprocess

engineering in solid waste management, Ph.D

thesis, Pondicherry University, Puducherry.

pp. 182.

3

Hendriksen, N.B., 1990. Leaf litter selection by

detritivore and geophagous earthworms. Biol.

Fert. Soils 10, 17-21.

Kumar, T.G., 2009. Vermicomposting of pernicious

weed salvinia (Salvinia molesta, Mitchell).

M.Phil thesis, Pondicherry University,

Puducherry. pp. 88.

Makhija, M., 2012. Vermicomposting of solid

waste: Leaf litter, ipomoea, and used paper.

Ph.D thesis, Pondicherry University,

Puducherry. pp. 106.

Marhan, S., Scheu, S., 2005. Effects of sand and

litter availability on organic matter

decomposition in soil and in casts of

Lumbricus terrestris L. Geoderma 128, 155-

166.

Schulmann, O.P., Tiunov, A.V., 1999. Leaf litter

fragmentation by the earthworm Lumbricus

terrestris L. Pedobiologia 439, 453-458.

Garg, V.K., Kaushik, P., 2005. Vermistabilization

of textile mill sludge spiked with poultry

droppings by an epigeic earthworm Eisenia

foetida. Bioresour. Technol. 96, 1063-1071.

Lim, S.L., Wu, T.Y., Sim, E.Y.S., Lim, P.N.,

Clarke, C., 2012. Biotransformation of rice

husk into organic fertilizer through

vermicomposting. Ecol. Eng. 41, 60-64.

Lim, S.L., Wu, T.Y., Clarke, C., 2014. Treatment

and biotransformation of highly polluted agro-

industrial wastewater from a palm oil mill into

vermicompost using earthworms. J. Agric.

Food Chem. 62, 691-698.

Shak, K.P.Y., Wu, T.Y., Lim, S.L., Lee, C.A.,

2014. Sustainable reuse of rice residues as

feedstocks in vermicomposting for organic

fertilizer production. Environ. Sci. Pollut. Res.

21, 1349-1359.

VERMICOMPOST CHARACTERISTICS AND ITS EFFECT

ON SOIL AND PLANT GROWTH – STATE-OF-THE-ART

Chapter

2

5

CChhaapptteerr 22

Vermicompost characteristics and its effect on soil and plant

growth – state-of-the-art

Abstract

The beneficial effects of vermicast on soil productivity and plant growth has been appreciated since the early

work of Hensen (1877) and Darwin (1881). In general, the castings have low carbon and nitrogen ratio, high

porosity, and good water-holding capacity than soil. Vermicast contains most nutrients in forms that are

readily taken up by the plants. It is also reported to consist of plant growth-promoting substances. The

potential of vermicast to improve the physical, chemical and biological properties of soil and the subsequent

effect of these changes on plant growth have also been reported in both natural and agroecosystems. All

these beneficial properties of vermicast are subject to various changes with substrate type, earthworm species

and age of castings. In this chapter, dynamics of all these properties of vermicast and its impact on plant

growth and soil is reviewed.

1. Introduction

The structure of the soils and the

biogeochemical cycles associated with the soils are

strongly influenced by soil invertebrates (Lavelle et

al., 2006; Barrios, 2007). Among the soil

organisms, earthworms are unarguably the most

important in maintaining the soil integrity by

several means in various vegetated lands (Edwards,

2004). Though they are not dominant numerically,

they represent a major invertebrate biomass in soils

due to their much larger size in comparison to the

two other most influential soil-working organisms –

the ants and the termites. The physical and

chemical characteristics of the casts produced by

earthworms plays an important role in the

regulation of soil processes. The physical structure

and chemical linkage of soil and feed are

reorganized during the gut transit that leads to

modifications in their porosity, density, and

hydraulic-properties (Shipitalo and Protz, 1989;

Jouquet et al., 2008a). The contents of the feed are

also modified by gut microbes and enzymes that

increase mineralization of organic matter, and

therefore increase the plant available form of

nutrients in castings (Chapuis-Lardy et al., 2010;

Bityutskii et al., 2012).

Although the role of vermicast in the

regulation of soil processes and plant growth has

been explored in detail, little is known on how the

properties of castings vary with earthworm‘s

species and its life-traits and the feed they ingest

(Schrader and Zhang, 1997; Buck et al., 1999; Jana

et al., 2010). Moreover, the life-time and the

disintegration of these structures, which are of great

importance in nutrients dynamics, are yet to be

explored in detail.

This chapter reviews the (i) potential factors

that determine the properties of vermicast, (ii)

change in the biological, chemical, and physical

attributes of the vermicast with time, and (iii) the

impact of the factors stated above on plant growth

and soil.

6

2. Types of vermicast and its composition

2.1. Types of vermicast

Earthworms mainly ingest feed rich in

organic matter together with living

microorganisms, nematodes and other microfauna,

mesofauna and their dead remains. Most species

also consume mineral soil fractions at various

degrees and indicate that they prefer organic–

mineral mixtures than pure organic materials

(Doube et al., 1997). Earthworms digest these

materials with the help of the gut associated

microorganisms and assimilate nutrients from them.

The vermicast excreted after gut passage are of

various forms and exist with different features. The

nature of the castings is determined by earthworm

species, their habitat and feeding behavior.

Generally, the earthworms produce following type

of biogenic-structures.

2.1.1. Based on place of deposition: surface and

subterranean castings

Casts may be deposited on the surface of the

soil or within it, depending on the species (Lavelle,

1988), food source, and bulk density of soil they

inhabit (Binet and Le Bayon, 1999). Based on the

place of deposition, the castings can be classified

into two groups – (i) surface castings and (ii)

subterranean castings. All the earthworms‘ species,

epigeic, anecic as well as endogeic deposit castings

at soil‘s surface layer, which has significant impact

on the physical and chemical characteristics of soil.

These surface casts increase the roughness of

surface soil and in turn affects water runoff and

infiltration into soil (Binet and Le Bayon, 1999; Le

Bayon and Binet, 2001). However, the fresh

castings are of low stability, easily dispersible in

runoff water, and lead to soil erosion and loss of

sediment-associated nutrients (Sharpley and Syers,

1976; Sharpley et al., 1979; Shipitalo and Protz,

1988; Binet and Tréhen, 1992; Ganeshamurthy et

al., 1998; Buck et al., 1999; Le Bayon and Binet,

1999). Moreover, surface casts dry out quickly,

harden, and, if compact, are likely to limit root

penetration, thereby reducing the ability of plant

roots to obtain the nutrients stored inside the casts

until they are broken down.

Anecic and particularly endogeic earthworms

produce subterranean castings, which are deposited

on the burrow walls, within the burrow or into other

sorts of soil macropores (Brown et al., 2000). The

subterranean castings do not influence the surface

soil properties such as nutrient transport, infiltration

and soil erosion (Jouquet et al., 2008a). Unlike

surface castings, the subterranean castings remain

fresh and retain moisture for longer duration and

are protected from the main agents of physical

degradation such as raindrops, animal trampling

etc. Hence, the degradation of these belowground

casts is probably even slower than for surface casts.

For instance, the half-lives ranged from 2 to 11

months for castings of larger anecic species,

Martiodrilus sp. (Binet and Le Bayon, 1999;

Decaens, 2000; Mariani et al., 2007). This species

produced both surface and subterranean castings

with an average dry weight of 25 g (Decaens,

2000). The casts of this species are likely deposited

more prior to their abandonment from borrows,

which is commonly the case after inundation by

heavy rainfall (Mariani et al., 2007). The surface

castings of this species were reported to be stable

for up to 4 months, and to then sharply fall after 6

months because of physical degradation of the casts

(Decaens, 2000). The castings of compact type,

inhibit the root extension and other hand, if they are

of the decompact type allow roots to penetrate more

easily and uptake nutrients available to plants

(Edwards, 2004).

2.1.2. Based on shape of castings

The shape of the castings produced by

different species is dissimilar; their properties and

the impact on soil may therefore differ considerably

(Lavelle, 1988). Castings can be classified into four

major categories based on their shape. They are: (i)

globular, (ii) paste likes slurry, (iii) tall vertical

7

columns or heaps of different shapes, and

(iv)granular (Edwards, 2004). Production of

castings of any of these types appears to be related

to soil texture and the size of earthworm. Specific

anatomic features of earthworms have also been

identified that allow the worm either to release

large units at regular intervals that will form the

globular cast, or a thin constant flux of cast material

that will form the poorly structured granular casts

(Lavelle and Spain, 2003). The shape and size of

the castings produced by some earthworm species

has been summarized in Table 1.

(i) Globular castings: Globular castings

consist of coalescent round or flattened units, which

are denser than the surrounding soil (Edwards,

2004; Blouin et al., 2007). This type of castings is

produced by anecic and endogeic species which are

larger species. It is stable for several months. The

greater structural stability of this castings is

probably due to the intense mixing and compaction

of gut content with mucopolysaccharides before it

released, which glue the soil particles together and

produce compact stable-structures (Lavelle, 1988).

Moreover, the castings are excreted with some

colorless waxy fluid from their nephridia (Agarwal

et al., 1958), calcium humate and calcite generated

during the organic matter decomposion and

calciferous glands, respectively which all perhaps

function as the cementing material to the castings

and harden them (Oyedelea et al., 2006). These

stable globular castings improve water infiltration

due to enhancement of the soil surface roughness

and modification in the circulation. Being water

stable aggregates, the vermicast is prevented from

detachment of easily transportable particles, which

in turn reduces erosion (Jouquet et al., 2008a).

An anecic earthworm species, Amynthas

khami in the Northern Vietnam (Bouché, 1977), is

reported to build biogenic structures up to 20 cm

length, which are formed from continuous

deposition of globular casts (Jouquet et al., 2008b).

These structures are connected with belowground

galleries and aboveground casts (Jouquet et al.,

2008a). Similarly, the large size globular castings

were reported in Andiodrilus pachoensis, a large

anecic earthworm. They deposit casts in the form of

piles on the soil surface. The piles can reach up to

12 cm height with a central canal called the

earthworm gallery (Thomas et al., 2008). Casts are

deposited on several occasions less than one week.

Hence the old and fresh portion of vermicast can be

differentiated. But once the formation of casts

diminishes, this heterogeneity disappears.

(ii) Slurry type of castings: Slurry type of

biogenic structures is single masses of soil mainly

produced by anecics and endogeics (Edwards,

2004). These casts are without a distinct shape,

paste-like and unstable, but stabilize with ageing

(Marinissen and Dexter, 1990). Martiodrilus sp. a

large size, dorsally dark-grey pigmented, surface-

casting species is reported to produce this type of

castings. The size of these casts is large. The casts

are of 3-6 cm diameter, height 2 to 10 cm and 25 g

average dry weight. The biogenic structures are

become tower like shaped by dry material at its

base and fresh pasty material at the top (Jiménez

and Decaëns, 2004).

(iii) Tall vertical columns or heaps of

variable shape castings: This type of castings is

usually deposited by anecic or endogeic groups.

These are formed by the sequential deposition on

globular casts. They have a hole in the middle when

they are tower form (Edwards, 2004). This type of

castings has been reported with Martiodrilus

carimaguensis. The castings are up to 15 cm height

with 5 cm diameter. They form tower like structure

which are deposited in the course of several days

(Jiménez et al., 1998; Decaëns et al., 1999).

Another species of earthworm Hyperiodrilus

africanus, which is a predominantly surface casting

species in large parts of moist savannahs of West

Africa, produced same type of castings. H.

africanus travels in the same channel several times

to deposit ingested soil at the soil surface, forming

characteristic turret shaped casts (Hauser and

Asawalam, 1998). Large amount of turret shaped

8

Table 1. Shape and size of the castings produced by different earthworm species.

Earthworm species Average size of the

worms

Average

weight of the

worms (mg)

Shape of the castings Range of the size of

castings References

Amynthas khami Up to 500 mm length - Globular 100- 200 mm height x 30

mm diameter

Jouquet et al., 2008a,b;

Podwojewski et al., 2008

Andiodrilus pachoensis length of 200 mm -

Globular and central

canal which corresponds to the

earthworm gallery

120 mm height Thomas et al., 2008

Dichogaster jaculatrix 10 or 25 mm length ×

1.4 mm diameter - Tower like structure

100 -120 mm height x 40

mm diameter Baylis, 1915

Drawida papillifer 60-120 mm length ×

3-5mm diameter 200-730

Composite irregular paste like

slurries

10-15 mm height × 8-10

mm diameter Chaudhuri et al., 2008,2009

Eudrilus eugeniae 90- 416.3 mm length ×

4-8 mm width 230- 5923

Fine granular pellets deposited

small heaps, that may be 20 to

30 mm high and 30-50 mm

diameter

30 mm height x 30-50

mm diameter

Madge, 1969; Viljoen and

Reinecke, 1994; Kale, 1998;

Parthasarathi, 2007; Mainoo et al.,

2008

Eutyphoeus assamensis 260-275mm length ×

5-6 mm diameter 1210-3730 Tower like structure

30-50 mm height × 15-30

mm diameter Chaudhuri et al., 2009

Eutyphoeus callosus 310-410 mm length ×

8-10 mm diameter 8200-15800

Tower like castings with

compact and thick convolutions

40-55 mm height × 60-65


Eutyphoeus comillahnus 120-165 mm × 3-5 mm

diameter 1000-2100

Tower like casts of fragile

aggregates with or without

convolutions

40-50 mm height ×25-30


Eutyphoeus gammiei 200-400 mm length ×

7-10 mm diameter 8350-13200



140-160 mm height × 40-

50 mm diameter

Bhattacharjee and Chaudhuri,

2002; Chaudhuri et al., 2008,

2009.

Eutyphoeus gigas 145-290 mm length ×

7-11 mm diameter 2230-5200



50-70 mm height × 40-50

mm diameter Chaudhuri et al., 2008, 2009

Eutyphoeus scutarius 244-332 mm length ×

6-7 mm diameter 4750-8180



35-70 mm height × 20-35


Eutyphoeus sp 80-195 mm length ×

3-4 diameter 500-1280

Tower like casts of fragile

aggregates with or without

convolutions

8-10 mm height × 20-25


Eutyphoeus turaensis 135-190 mm length ×

2.5-3 mm diameter 400-1160


compact and thin convolutions

30-35 mm height × 10-25


9

Earthworm species Average size of the

worms

Average

weight of the

worms (mg)

Shape of the castings Range of the size of

castings References

Hippopera nigeriae - -

Pipe like composite cast, which

had an average bore diameter of

2.0 mm ranging from 0.9-3.6

mm

16-118 mm height x 1.5

mm diameter

Nye, 1955; Wilkinson and Aina,

1976.

Hyperiodrilus africanus 120-160 mm length -

Pipe like composite cast, with a

vertical hole running through it

but closed at the top.

25-90 mm height x 10-

30 mm diameter

Madge, 1969; Lee, 1985; Hauser

and Asawalam, 1998; Tondoh,

1998.

Kanchuria sp 175-280 mm length ×

2.5-6 mm diameter 1550-3450 Large globoid

20-45 mm height × 30-45

mm diameter Chaudhuri et al., 2008, 2009.

Martiodrilus

carimaguensis

and 194.3 mm in length

and 9.3 mm in diameter

11200 Tall vertical heaps (turricule) Up to 150 mm height x

up to 50 mm diameter

Jiménez et al., 1998; Decaëns,

2000; Tondoh and Lavelle, 2005;

Mariani et al., 2007a; Mainoo et

al., 2008.

Metapheretima

jocchana

500-670 mm length ×

9-10 mm diameter - Very large composite casts

50 mm height x 50 mm

diameter Lee, 1967; Tsai et al., 2004.

Metaphire houlleti 92-200 mm length ×

4-7 mm diameter 1000-3130

Tower-like casts with regular

arrangement of

spherical/subspherical

aggregates

35-70 mm height × 15-35


Metaphire posthuma 120 mm in length ×

5 mm diameter -

Granular , which are mainly

found belowground 2 to 3 mm diameter Bottinelli et al., 2010

Microchaetus sp 1,800 mm length ×

16-18 mm diameter - Turret shaped

30-100 cm height and 1

m diameter

Edwards and Bohlen, 1996;

Henrot and Brussaard, 1997;

Blakemore et al., 2007.

Notoscolex birmanicus

760-1200 mm length ×

5 mm diameter

- Tower like structure 200 -240 mm height x up

to 40 mm diameter

Gates, 1961; Tembe and Dubash,

1961; Blakemore et al., 2007.

Perichata sp - - Tower like structure 90 mm height x 40 mm

diameter Darwin, 1881

Pontoscolex

corethrurus

50-150 mm length ×

4-6 mm diameter

350-800

Composite irregular paste like

slurries

10-15 mm height × 10-15

mm diameter

Moreno and Paoletti, 2003;

Dlamini and Haynes, 2004;

Chaudhuri et al., 2008, 2009.

10

casts deposition is also reported in Microchaetus

microchaetus (Henrot and Brussaard, 1997).

Usually many endogeics deposit casts for several

days in the same place as they exhibit restricted

movement and thus build spectacular tower-like

structures. Casts are piled up to 5-10 cm height in

the moist tropical forests of Africa. This is common

in many other tropical regions also (Lee, 1985).

Lavelle (1968) has reported similar, but less

spectacular piles at Lamto. These casts were

deposited by small Eudrilidae species continuously

for up to 30 hours at the same place (Lavelle,

1968).

(iv) Granular castings: Granular castings are

pellet like. They are produced mainly by smaller

earthworm species (epigeic, small endogeic, and

some anecic species) and distributed on or beneath

the soil surface (Edwards, 2004). The pellets are

small fine-textured, unlike globular castings and

hence are subjected to wash away by rain more

easily. Therefore, it favors sheet erosion (Lavelle,

1988). Examples for the granular casts depositing

earthworms are Eudrilidae sp. and Eudrilus

eugeniae, which produce castings of < 5 mm

diameter in size (Mulongoy and Bedoret, 1989;

Henrot and Brussaard, 1997; Hauser et al., 1998).

2.1.3. Based on size of the castings

Based on size, the castings can be categorized

into two types: (i) large size (>5mm) castings and

(ii) small size (<5 mm) castings. The large size

castings are egested by species of compacting

earthworm. They produce castings of higher bulk

density than the soil they inhabit. The high bulk

density is due to: (i) the formation of organo-

mineral bonds after mixing and chemical

transformations in the gut; (ii) the reabsorption of

water in the latter part of the earthworm intestine;

and (iii) the strong compaction by the tail muscles

when casts are expelled (McKenzie and Dexter,

1988). Casts of compacting species are stable in

nature as it creates anaerobic conditions inside the

castings which slows down the microbial

decomposition (Blanchart et al., 1993). The small

size castings are formed by decompacting

earthworms. They are of low bulk density. The

castings of decompacting earthworms are not stable

as that of the castings produced by compacting

species.

The larger size castings may be globular,

paste like slurries, tall vertical heaps or column

shaped castings, and small size castings is granular.

The production of both the types of casts by a

different earthworm group is favorable for

maintenance of the soil structure (Decaëns, 2000).

For example, in tropical agroecosystems, large

endogeics, such as Pontoscolex corethrurus or

Millsonia anomala, egests compact casts which are

large in size. These castings increase the proportion

of macroaggregates and bulk density of soil. This

cast might create anaerobic conditions inside the

castings and in turn slow down the decomposition

(Blanchart et al., 1993). Smaller decompacting

earthworm species (e.g. Eudrilidae) feed on these

large casts and consequently produce small and

unstable casts (0.5–5 mm) (Blanchart et al., 1999).

When decompacting species ingest casts of

compacting species, previously physically protected

carbon gets mineralized. On the other hand, where

there is ingestion of casts of decompacting species

by compacting species, the mineralizable carbon it

contains tends to be protected in their large casts

(Dickschen and Topp, 1987; Blanchart et al., 1999).

The activity of compacting earthworm species

alone lead to the formation of a compact surface

crust (Six et al., 2004).

The size of the castings has strong positive

correlation with the size of earthworm species that

produce them (Brussaard and Ferrera-Cerrato,

1997). The size of few earthworm species and the

size and shape of the castings that they produced is

listed in Table 1. Usually the size depend on the

species and range between 1-10 mm in diameter

(Oades, 1993). The sizes of the castings also may

be varying within same species, which rely on the

feed quality (Jiménez et al., 1998). For instance,

11

Martiodrilus sp, a large anecic species in the

Eastern Plains of Colombia, exhibit high variability

in their diet (Mariani et al., 2007). They produce

casts with low organic content but larger in size

(Jiménez et al., 1998). This is due to the availability

of low nutrient containing substrate as feed to

earthworm. As a result they ingest more quantity of

feed to fulfill their energetic requirements (Curry

and Schmidt, 2007). The large size of casts may

also influence oxygen-diffusion into them; the

mineralization of nitrogen, nitrification and oxygen

uptakes are slower in larger casts than the smaller

aggregates (Adu and Oades, 1978). Studies of

Sextone et al. (1985) also support this finding, in

which the oxygen content of a wet aggregate (18

mm diameter) near the aggregate center is 0% and

at the edge of the aggregate 21%.

2.2. Physical properties of vermicast

2.2.1. Color and odor

The color of the castings is a main factor to

distinguish the castings from the soil and other

components in field and vermicomposting process

(Lavelly, 1968; Garcìa and Fragoso, 2002). The

casts are completely dissimilar from the feed being

generally dark brown to black and mull humus-like

in appearance (Hervas et al., 1989; Brown et al.,

2000; Barros et al., 2001). Dramatic changes in

color of these castings are reported during aging.

Binet and Le Bayon (1999) reported three different

color castings in temperate cultivated soils. They

are brown, light brown and whitish. Those castings

may be produced from six different earthworms‘

species, reported in their study area. The brown

type is fresh casts, brown light are moderate aged

dry castings and whitish castings are oldest eroded

type. Depending on the climatic conditions the

casts may be covered by moss of white fungi giving

whitish appearance to the aged and disintegrated

castings (Binet and Le Bayon, 1999). Castings from

different horizons also show variation in their color.

The castings of Pheretima darnliensis derived from

the three different horizons showed three different

colors. The castings of P. darnliensis was dark

brown and grey-yellow when it was derived from O

and A horizon, respectively and castings of

intermediate color was also produced which

resembled a mix of the two soil layers (Gould et al.,

1987).

The vermicast has been reported to offer faint

pleasant earthy odor (Senesi, 1989; Szczech, 1999;

Dickerson, 2001; Sharma et al., 2005; Turnell et

al., 2007), may be due to the inhibition of the

growth of foul-smell forming microbes in the

decaying waste biomass when passed through

earthworm gut (Pierre et al., 1982). The anti-

microbial property of secretion of pharyngeal

glands, intestine, gizzard and crop, may also be the

reason for modulation of the diversity of the gut

microflora (Edwards and Fletcher, 1988). The

antibacterial factors of earthworms are also

reported in the coelomic fluid of Eisenia fetida

(Lassegues et al., 1989; Valembois et al., 1991) and

Lumbricus terrestris (Tuckova et al., 1986;

Anderson, 1988). Earthworms also create aerobic

conditions in the waste materials, prohibiting the

function of anaerobes which produce mercaptans

and hydrogen sulfide which gives foul odour

(Pedersen and Hendriksen, 1993; Sinha et al.,

2002).

2.2.2. Bulk density

The bulk density of castings also play an

important role regardless of nutrients they contain,

influencing the density and hydrological properties

of soil they exist. In general, soil with low bulk

density (0.7–1.8 g cm-3

) is desirable for plant

growth (Lal and Shukla, 2004), whether those are

agricultural crops, trees, or turf grass. Moreover,

soils of low bulk density have a greater water

infiltration rate which reduces storm water flow and

runoff. On the other hand, a high bulk density can

impede root penetration and reduce the air and

water circulation in the soil. Since the amount of

cast production is high on surface, its effect on bulk

density of surface layer is highly significant. The

12

amount of castings deposited by earthworms over

the soil surface may range from 2 to 50 tonnes per

year, and it may exceed 1000 tonnes per year when

the belowground castings are included, which rely

on the type of soil and land use (Lee and Foster,

1991). Ponomareva (1950) and van de Westeringh

(1972) have referred that earthworm casts

constitute up to 50% of soil aggregates in surface

layer in temperate pastures. The top layer of mull-

type forest (Kubiena, 1953) wooded savanna in

Ivory Coast (Lavelle, 1978), consisted of almost

entirely earthworm casts; whereas, James (1991)

estimation shows that surface castings produced in

Kansas tallgrass prairie is equivalent to 4-6% of

surface at the top 15 cm soil each year.

The bulk density of the castings greatly vary

with different earthworm species. Usually the bulk

density of castings produced by compacting species

is higher than of the soil they inhabit. Introduction

of a compacting species, Millsonia anomala in a

de-faunated savanna in Ivory Coast by temporary

flooding, increased the soil bulk density much

higher than of its initial within 20 months period

(Lavelle and Spain, 2003). As mentioned in section

2.1.3, the factors that increase the bulk density: (i)

the mixing and chemical transformations in the gut

which facilitates the bonding of organic matter and

mineral constituents; (ii) the reabsorption of water

in the latter part of the earthworm intestine; and (iii)

the strong compaction by the anus when casts are

ejected (McKenzie and Dexter, 1988). The smaller

de-compacting species produce smaller and fragile

castings, with lower bulk density due to the less

dense packing when castings are expelled (Six et

al., 2004). The micromorphological observation on

casts from the de-compacting species also showed

soil particles less densely packed and with a high

fraction of bacteria than those of compacting type

(Josehko et al., 1989). The bulk density of fresh

castings of de-compacting species, Aporrectodea

rosea was about 1.15 mg cm -3

, when it bred in a

sandy loam soil (McKenzie and Dexter, 1987). It is

much lower than the bulk density of Millsonia

anomala, a compacting species in Lamto‘s

savannas of Ivory Coast, which produced castings

with high bulk density of 1.8 mg cm−3

, when the

bulk density of surface soil was about 1.45 mg

cm−3

. A high sand content (75%), with the low clay

(7.5%) and organic carbon (1%) also might be

reason for the higher bulk density of castings of M.

anomala (Blanchart et al., 1993).

The bulk density of the castings may vary

with different soil type. For instance, earthworm

casts of temperate ecosystems have bulk density

lower than the surrounding soil (Larink et al., 2001;

Marashi and Scullion, 2003) while those from

tropical soils are denser than the former (Blanchart

et al., 1993, 1999; Decaëns et al., 2001; Oyedele et

al., 2006). Bulk density of an anecic, Martiodrilus

carimaguensis, in a native savanna was 10% higher

than the castings of the same species in an intensive

pasture. The higher proportion of organic matter in

the casts deposited in the pasture may be the reason

for the lower bulk density when compared to casts

of the savanna (Decaëns, 2000). Similarly, the bulk

density of castings produced by Lumbricus

terrestris was 1.33 and 1.54 mg cm-3

in sandy silt

and loamy silt soil, respectively (Josehko et al.,

1989). The difference in the bulk density of these

castings was probably due to their varying organic

carbon content. The organic residues have been

shown to buffer the effects of compaction as they

possess the ability to be compressed and then

springs back to its normal shape once the pressure

has released. This property of organic matter

prevents the compaction of clay fraction of soil

which is attached with organic matter. Therefore,

high organic matter levels make the soil less

susceptible to soil compaction (Wortman and Jasa,

2003). As the castings age, the bulk density either

decrease or increase, which depends on the species

which produced them.

13

2.2.3. Pore space

Porosity is a major constraint on other

physical, chemical and biological properties; it is of

primary importance in the regulation of tensile

strength, hydrological properties, gas diffusion,

microbial colonization, nutrient mineralization etc.

The earthworms alter the soil pore space through

their borrowing and casting activities (Edwards,

2004). The burrowing activity of earthworms

substantially alters soil macro pores (>1 mm),

which act as flow paths in soil (Bouche´ and Al-

Addan, 1997; Trojan and Linden, 1998). The

castings deposited affect mainly the meso- and

microporosity in soil. During the gut transit, the

pore space between organic and mineral particles

may get altered and creates packing void in the

deposited castings (Shipitalo and Protz, 1989;

Blanchart et al., 1993; Chauvel et al., 1999). Casts

has rather low amount of tubular, crack and almost

no packing void poroids as compared to the soil

(Blanchart et al., 2004; Jouquet et al., 2008b). The

structural rearrangement of the soil particles in the

intestine of earthworm would result in more fine

pores and fewer macropores in the castings

(Blanchart et al., 1993). These pores can be divided

into three size classes: (i) transmission (>50 µm),

(ii) storage (0.5 to 50 µm) and (iii) residual pores

(<0.5 µm). The storage and residual pores are

mainly found in the casts while the transmission

pores are located between the casts (Decaens et al.,

1999b). These transmission pores control the

transport of water and solutes and (Lamande et al.,

2003) the storage pores retain water with greater

tenacity particularly in the pores in the cortex of the

casts which are of 1-10 µm size (McKenzie and

Dexter, 1987).

All these three classes of pore space vary

considerably based on the earthworm species that

produce them, particularly the structure of

earthworm‘s anterior and posterior musculature

(Lapied and Rossi, 2000). In general, the casting of

de-compacting species contains more meso-pores

than those of compacting species. Even the pore

space in the castings of similar type may vary

significantly. For instance, the castings of a

compacting species, Millsonia anomala contain

predominantly mesopores ranging between 10 to 20

µm, whereas, the castings of a similar compacting

species, Pontoscolex corethrurus were all smaller

than 1 µm (Chauvel et al., 1997). Due to the results

of fewer meso-pores, the compacting species

reduced infiltration rates (Blanchart et al., 2004).

The pore space in the castings may greatly

influence the structure, and function of the

microorganisms and microfaunal communities and

interaction between them. Also the pores of

different size support different microbial

populations (Hattori, 1988). For example, pore

diameter larger than 90 µm favor the growth of

microarthropods, whereas, it is not supported in

smaller than 1.2 µm diameter pores (Vreeken-Bujis

et al., 1998). Pore sizes in the order of 1 µm

support the growth of bacterial population, and the

pore diameter of 30-90 µm enhance the nematode

population density (Hassink et al., 1993). The

greater percentage of pore size of casts of compact

earthworm species Millsonia anomala (Blanchart et

al., 1993) were less than 10 µm, which impedes

nematode colonization and predation (Elliott et al.,

1980). Visser (1985) and Anderson et al. (1984)

suggested that fungal growth is greatest in soils

with large pore sizes that can allow fungal

sporulation (Vreeken-Bujis et al., 1998). The

growth of protozoan population may also be

influenced by structural limitations, since the

spatial location of the bacterial population influence

the effectiveness of certain species (free swimming

and larger) (Clarholm, 1981; Vargas and Hattori,

1986; Kuikman et al., 1990). On the other hand, the

castings produced from the Lumbricus terrestris

and Aporrectodea caliginosa are favorable habitat

for protozoa, due to their higher (about 10%)

porosity than soil. Castings of Octolasion lacteum

also shows higher protozoan population than the

soil aggregates (Bonkowski, 1995). Apart from

high microbial biomass which is source of feed of

protozoans in the cast, the pore and its distribution

14

also improve the predation (Larink et al., 2001).

Oxygen concentration will also control the kind and

distribution of organisms which in turn is affected

by pore space in the castings (Elliott and Coleman,

1986).

As the changes in the pore space greatly

modify the structure of microbial community,

mineralization and immobilization of nutrients also

vary significantly as it is partially controlled by the

size, and composition of microorganisms and their

activity (Marschner and Kalbitz, 2003; Fontaine

and Barot, 2005). The mineralization of organic

matter may be limited by pore size distribution due

to the positioning of organic matter in pores which

restricts the access to the microorganisms, thereby

causing restricted predation of those

microorganisms (Elliott and Coleman, 1988;

Hassink, 1992; Ladd et al., 1993; Strong et al.,

2004). The protozoa are found in pores > 6 µm dia

in which higher C mineralization is reported than

the bacteria populated pores of < 6 µm (Killham et

al., 1993). Moreover, the pores of different size

control the exchange of water and gases, which in

turn influence the availability of organic substrates

to the organisms. For instance, in the pores of

diameter in the range of 6-30 µm, the

mineralization of C was high when these pores

were filled with water and which attributed the

availability of microbial biomass to protozoans in

these pores (Killham et al., 1993). Increasing the

proportion of water-filled porosity increases the soil

water potential which consequently increases

mineralization rates (Sommers et al., 1980) but it

may create anaerobic conditions and stimulate

denitrification when the water-filled pores are more

than 60% of total pore space (Linn and Doran,

1984; Bergstrom and Beachamp, 1993; Elliott and

de Jong, 1993).

2.2.4. Water-related properties

As the earthworms casting activity has

significant impact on the soil porous system, soil

hydraulic functions such as infiltration, erosion,

runoff, water retention and evaporation are also

strongly influenced (Lamandé et al., 2003;

Bottinelli et al., 2010). The packing voids within

cast change the infiltration rate by transporting and

retaining large amounts of water and solutes

through it (Lamandé et al., 2003), which plays

significant role in regulating soil quality. Sufficient

amount of water need to infiltrate into soil and soil

aggregates, because it may create anaerobic

conditions when it is restricted that slows down the

organic matter decomposition (Elliot et al., 1990;

Blanchart et al., 1993). Moreover, lower soil

infiltration rate is generally correlated with decline

in soil quality due to increase in runoff and soil

erosion (Piekarz and Lipiec, 2001; Bouchb and Al-

Addan, 1997).

The effects of earthworms on infiltration may

vary with ‗compacting‘ and ‗decompacting‘ species

as they produce castings of different size and

porosity (Hallaire et al., 2000). Studies conducted

on different soil systems have shown that the

castings of Pontoscolex corethrurus, a compacting

species led to significant change in structure of

surface soil. The outermost layer became highly

compacted due to numerous compact structures

which caused impermeability and reduced the

infiltration (Alegre et al., 1995; Barros et al., 1996;

Young et al., 1998; Chauvel et al., 1999; Hallaire et

al., 2000). Castings deposited by other compacting

species, Millsonia anomala has also been reported

to impede infiltration (Spain et al., 1992; Derouard

et al., 1997), in both laboratory and field studies.

The high bulk density and low mesoporosity of

compacting type castings may probably be the

reason for impeding infiltration (Blanchart et al.,

2004).

In contrast, larger size compacting type

castings produced by Amynthas khami are reported

to improve water infiltration by enhancing

roughness of the surface soil, modification in the

circulation of surface runoff and reducing the

velocity in steep-slope ecosystems of Northern

Vietnam (Jouquet et al., 2008). The differential

15

impact of castings produced by A. khami probably

due to its larger size which has large packing voids

allows a large amount of water to infiltrate.

Moreover, these castings are anchored in soil, and

are connected with below-ground galleries which

facilitate the infiltration (Jouquet et al., 2008).

However, the fresh and decompacting type castings

are highly dispersible in water, and clog the

transmission pore, thereby impeding infiltration of

water in soil (Shipitalo and Protz, 1988; Le Bayon

and Binet, 1999). A similar, impact on soil

infiltration was reported with the castings of

Aporrectodea tuberculata and Metaphire posthuma.

Although, the castings of these species initially

increased the infiltration rate due to their high

porosity, the unstable nature of this structure led to

rapid compaction of soil after rainfall or overland

flow (Ela et al., 1992; Binet and Le Bayon, 1999;

Bottinelli et al., 2010). However, balance between

the amount of cast deposited over the soil surface

and its degradation rate regulates the soil structural

porosity, which in turn influence infiltration. Soil

structural improvement would occur when the

amount of cast production predominates and the

effect would be reversed when degradation of these

structures was more (Bottinelli et al., 2010). In

addition, the ageing of castings also influence their

impact on soil. In general, dispersion of fresh

castings is high, and it becomes stable during

ageing under several drying–wetting cycles

(Decaëns et al., 1999b,c; Mariani et al., 1999). The

stable aggregates can protect from detachment by

rainfall or surface flow, and reduce the velocity of

surface runoff and as a consequence increase

infiltration.

In addition, the amount of clay present in the

castings is also one of the main soil parameter that

affect the soil surface sealing and infiltration

(Shainberg and Levy, 1996), as the increase in clay

content increases with increase in the stability of

aggregates. This is especially due to the

aggregation and bonding effect of clay (Le

Bissonnais, 1996). Thus, the stability of aggregates

against raindrop impact generally increases with an

increase in clay content (Blanchart et al., 2004).

Like the clay percentage, presence of organic

matter also affects infiltration because it is an

important stabilizing factor of aggregates (Shipitalo

and Protz, 1988). Many studies conducted in the

tropics have showed that most earthworms prefer

organic particles, depending on the species and

availability of organic content in soil (Barois et al.,

1999). As a result of difference in the behavior, the

stability of the castings they produce also vary, and

thereby its impact on soil infiltration may also be

different (Tomlin et al., 1995). The nature and

amount of exchangeable cations such as Ca2+

, Mg2+

,

K+, Na

+ and carbohydrates and lignin content of

plant debris present in the castings are reported to

influence erosion and infiltration through their

impact on dispersion and flocculation of clay (Le

Bissonnais, 1996), which may vary with different

earthworm species and substrate which is available

to them (Pop et al., 1992; Salmon and Ponge, 1999;

Oyedele et al., 2006).

2.3. Chemical properties of vermicast

2.3.1. Organic carbon

Carbon is a basic unit of all life form on

earth. Though not a nutrient element‚ it is the

building block of all organisms. Carbon is the

center for biological energy transfer within the

biosphere at landscape and ecosystem levels‚ and

within the organisms (Lavelle and Spain, 2003). In

the soil the effects of earthworm on carbon

dynamics vary, depending on the scale of space and

time that is considered (Lavelle et al., 1998). In the

short-term, earthworms enhance the carbon

mineralization of soil during their gut passage

(Blair et al., 1994); whereas, this effect is opposite

in aged castings in which carbon is protected from

further breakdown (Decaëns et al., 1999a; Kooch

and Jalilvand, 2008). Few studies revealed that

stabilized organic matter in the casts, can be

protected from microbial mineralization for many

years (Mcinerney and Bolger, 2000). The increase

in carbon stock in soil is gaining importance, as the

16

rise in CO2 and global warming are major concerns

(Bossuyt et al., 2005).

Although, castings produced by earthworms

stabilized the carbon for longer period, a

considerable amount of carbon loss through their

mucus secretion exceeds the carbon lost by their

respiration (Scheu, 1991). Intestinal mucus is

composed of amino acids of low-molecular weight

(about 200 Da) with carbohydrates and high

molecular weight glycoproteins (40000–60000 Da)

(Martin et al., 1987). Studies on 14

C and 15

N labeled

feed with Nicodrilus longus have shown that the

entire C content of the earthworm tissues could

turnover in 40 days, and a considerable portion of

this turnover was due to mucus excretion (Ferriere

and Bouche, 1985). The total C content of the

intestinal mucus of different earthworm species

ranged from 39–44 % C and 7–7.3 % N and it

seems to be similar across different species and

ecological categories (Martin et al., 1987). In the

study conducted by Scheu (1991) with geophagous

earthworm Octolasion lacteum, 63% of total C

losses (mucus excretion plus respiration) reported

was from secretion of mucus in casts and from the

body wall. This corresponds to a daily loss of 0.7%

of total C for this species, on the other hand,

respiration accounted for only 37% of total C loss.

Pontoscolex corethrurus secreted up to 50 Mg

mucus/ha in a single year in tropical pastures of

Mexico, which corresponds to 20% of the total C in

the soil (Lavelle, 1988).

The C in earthworm casts differ in form and

amount from those of the adjacent soil. Many

studies showed about 1.2 to 2 folds higher carbon

content in the castings compared to bulk soil (Lee,

1985; Shipitalo and Protz, 1988; Daniel and

Anderson, 1992; Barois et al., 1993; Zhang and

Schrader, 1993; Buck et al., 1999; Bossuyt et al.,

2005). The higher carbon content of castings are

probably due to their preferential ingestion of

organic rich residues such as leaf and root litter,

microorganisms, fecal pellets of other invertebrates,

etc. (Curry and Schmidt, 2007). It is also postulated

that the higher carbon content of castings is

probably due to the incomplete absorption of

carbon during the gut transit (Shaw and Pawluk,

1986; Daniel and Anderson, 1992). In contrast,

when the earthworms feed upon phytomass or

manure, there was drastic reduction in carbon

content in the egested castings. Aira and

Domínguez (2009) reported about 40% reduction in

C content of cow and pig manure after the transit

through the gut of Eisenia fetida. During the

digestion, absorption of C and microbial respiration

in the form of CO2 by worms reduces the C content

in the castings as compared to the feed. The

mixture of industrial lignocellulosic waste with

other organic supplements (saw dust and cow dung)

fed Perionyx sansibaricus produced castings that

exhibited a decrease in organic C content of 5.0 to

11.3% (Suthar, 2007). Similar C losses were

reported in casts of Eudrilus eugeniae fed with pig

manure (Aira et al., 2006). Contradiction in the

results is due to the preferential ingestion of high

organic matter by earthworm in field, and however,

C content of castings compared with only the bulk

soil. Therefore, earthworm castings show higher

amount of C content, still they assimilate

considerable amount C during gut transit. The gut

passage reduces by as much as 19% C in the soil

which they ingest (Edwards, 2004).

The C content of the castings vary

significantly depending on the earthworm species,

their feeding habits, particularly the amounts of

plant litter they intake (Table 2). Earthworms are

generally classified into three groups, epigeic,

anecic and endogeic (Bouché, 1977). Among them,

the epigeic and anecic species often behave as

litter-feeders, while the endogeics are geophagous,

consuming enormous quantities of soil in order to

meet their organic matter needs. Therefore, the

endogeic species are further divided into three

groups: oligohumic, mesohumic and polyhumic

(Lavelle, 1981). Based on the degree of food

selection, nutrient concentration is generally higher

in the casts in detritivorous than in geophagous

species (Buck et al., 1999). However, feeding

17

Table 2. Some of the chemical properties of castings produced by different earthworm species from different organic wastes.

Substrates Earthworm species Carbon mg g

-1 Nitrogen mg g

-1 Phosphorus mg g

-1 Potassium mg g

-1

References Cast feed Cast feed Cast feed Cast feed

Agricultural waste Eisenia foetida 35.0 106.0 8.8 1.8 1.0 0.70 2.63 0.62 Garg et al., 2006a

Buffalo dung Eisenia foetida 267.0 519.0 7.80 5.60 5.80 5.00 0.65 1.07 Garg et al., 2006b

Camel dung Eisenia foetida 407.0 463.0 5.20 4.00 3.20 3.00 0.36 0.50 Garg et al., 2006b

Coffee pulp Eudrilus eugeniae 147.0 535.0 16.6 10.3 4.10 1.30 7.00 1.20 Raphael and

Velmourougane, 2011

Coffee pulp Perionyx ceylanesis 168.0 535.0 18.6 10.3 5.10 1.30 7.80 1.20 Raphael and

Velmourougane, 2011

Couch grass (Cynodon dactylon) Eisenia fetida 342.1 497. 9 15.8 7.33 9.00 4.30 10.5 5.86 Pramanik et al., 2007

Cow dung Lampito

mauritii 268.7 298.9 7.73 5.78 2.73 2.32 6.00 5.42 Suthar, 2008

Cow dung Eisenia fetida 326.0 362.2 17.2 6.59 12.7 5.16 11.4 6.39 Pramanik et al., 2007

Cow dung Perionyx excavatus 201.6 285.9 19.3 12.8 8.13 5.72 9.51 4.93 Suthar, 2009

Cow dung Perionyx sansibaricus 200.2 285.9 20.4 12.8 8.35 5.72 9.66 4.93 Suthar, 2009

Cow dung Eisenia fetida 320.0 486.0 31.4 8.7 13.4 8.70 7.80 5.50 Yadav and Garg, 2011

Cow dung E. fetida + L. mauritii 261.7 298.9 9.18 5.78 2.77 2.32 6.15 5.42 Suthar, 2008

Dairy sludge + cattle manure Eisenia andrei 225.0 303.0 17.0 11.0 7.70 7.30 7.60 25.0 Elvira et al., 1998

Digestate from biogas plant Perionyx excavatus 285.0 364.0 40.0 18.0 14.0 9.00 10.4 6.50 Rajpal et al., 2014

Digestate from biogas plant Perionyx sansibaricus 276.0 364.0 40.0 18.0 13.0 9.00 10.0 6.50 Rajpal et al., 2014

Donkey dung Eisenia foetida 381.0 485.0 6.80 5.00 5.50 5.00 7.30 13.1 Garg et al., 2006b

Duckweed residues Eisenia fetida 261.5 445.8 13.1 6.24 9.0 3.96 10.5 5.72 Pramanik et al., 2007

Filter mud + saw dust Eisenia foetida 349.0 400.3 22.8 16.0 24.7 12.6 9.60 4.40 Khwairakpam and

Bhargava, 2009

Filter mud + saw dust Perionyx excavatus 357.0 400.3 24.4 16.0 22.9 12.6 7.50 4.40 Khwairakpam and

Bhargava, 2009

Filter mud + saw dust Eudrilus eugeniae 363.0 400.3 28.0 16.0 24.4 12.6 8.40 4.40 Khwairakpam and

Bhargava, 2009

Food industry sludge + poultry

droppings + cow dung Eisenia fetida 330.0 400.0 29.4 18.0 11.9 8.90 5.50 4.00

Yadav and Garg, 2011

Goat dung Eisenia foetida 231.0 438.0 8.90 4.70 4.70 3.70 3.40 7.20 Garg et al., 2006b

Guar gum industrial waste + cow

dung + saw dust Perionyx sansibaricus 431.7 481.9 18.3 15.4 3.90 2.50 13.2 11.5 Suthar, 2007

Horse dung Eisenia foetida 216.0 484.0 7.70 3.50 9.60 7.00 3.80 7.80 Garg et al., 2006b

Institutional waste Eisenia foetida 32.0 55.0 7.30 1.40 0.87 0.40 0.72 0.35 Garg et al., 2006a

18

Substrates Earthworm species Carbon mg g

-1 Nitrogen mg g

-1 Phosphorus mg g

-1 Potassium mg g

-1

References

Cast feed Cast feed Cast feed Cast feed

Kitchen waste Eisenia foetida 33.0 73.0 11.0 2.50 1.80 1.30 4.36 0.87 Garg et al., 2006a

Municipal solid wastes Eisenia fetida 242.6 200.3 7.62 3.74 5.10 3.52 7.00 4.28 Pramanik et al., 2007

Paper mill sludge + cattle

manure Eisenia andrei 151.0 266.0 12.0 11.0 5.90 3.90 7.60 23.0 Elvira et al., 1998

Poultry droppings + cow dung Eisenia fetida 350.0 420.0 28.5 11.9 12.9 9.00 6.10 4.30 Yadav and Garg, 2011

Press mud + cow dung Perionyx ceylanensis 290.0 544.1 16.3 9.40 23.8 15.0 31.3 21.2 Prakash and Karmegam,

2010

Sewage sludge Eisenia fetida 219.3 267.6 30.0 26.8 47.7 43.7 5.59 5.10 Suthar, 2010

Sewage sludge+ sugarcane trash Eisenia fetida 218.5 320.4 29.3 23.5 28.9 25.4 7.10 6.30 Suthar, 2010

Sheep dung Eisenia foetida 212.0 323.0 7.80 3.70 5.10 3.10 4.50 7.00 Garg et al., 2006b

Spent mushroom compost E. foetida + E. andrei 38.4 37.9 29.5 28.3 22.9 12.7 14.6 8.70 Tajbakhsh et al., 2008

Spent mushroom compost +

fruits and vegetables residues E. foetida + E. andrei 45.3 43.9 25.8 21.1 44.9 7.50 34.1 9.14 Tajbakhsh et al., 2008


pomegranate residues E. foetida + E. andrei 43.3 46.5 28.9 22.2 34.3 7.60 29.5 7.19 Tajbakhsh et al., 2008


potato residues E. foetida + E. andrei 81.1 45.9 21.4 22.1 36.8 7.95 81.9 13.7 Tajbakhsh et al., 2008


stump residues E. foetida + E. andrei 13.4 28.9 19.5 14.6 24.1 9.05 27.4 10.6 Tajbakhsh et al., 2008


tomato residues E. foetida + E. andrei 33.4 46.5 4.80 34.9 35.6 8.95 37.5 5.54 Tajbakhsh et al., 2008

Textile industry fibre waste Eisenia foetida 38.0 64.0 0.52 1.00 0.60 0.26 2.47 0.82 Garg et al., 2006a

Textile industry sludge Eisenia foetida 34.4 51.0 7.00 1.20 0.26 0.04 0.43 0.30 Garg et al., 2006a

Textile mill sludge+ cow dung Eisenia foetida 170.0 297.0 9.20 3.50 4.10 3.70 2.30 4.40 Kaushik and Garg, 2003

Winery industry waste Eisenia andrei 418.0 546.0 14.0 15.6 4.97 2.23 18.2 20.8 Nogales et al., 2005

19

behavior of earthworms also vary with different soil

type where they inhabit. Barois and Lavelle (1986)

demonstrated that the tropical species Pontoscolex

corethrurus was able to selectively ingest either

small mineral particles or large organic debris,

depending on the type of soil. Selection was made

on aggregates rather than primary particles.

However, some earthworms may selectively ingest

coarser particles than the average in soils with very

high clay contents. As a consequence, the C content

of castings also varies with species involved and

place they inhabit.

Moreover, the digestive systems of

earthworms of different ecological category have

different assimilation efficiencies, which also could

be the reason for differential carbon content of their

castings. Studies of Jegou et al. (1998) had shown

castings of Eisenia andrei (epigeic), Lumbricus

terrestris (epi-anecic) are enriched with organic

matter compared with bulk soil, whereas, those of

Aporrectodea giardi (anecic) and Aporrectodea

caliginosa (endogeic) contains lesser C than their

surrounding soil. The castings of epigeic and

epianecic species contained more than 50% litter C

(13

C labeled), while the anecic and endogeic were

of soil. Studies conducted in absence of leaf litter

showed that the C content of castings from the

geophagous Allolobophora chlorotica exceeded

that of Lumbricus rubellus, Lumbricus terrestris

and Octolasion cyaneum in soil without leaf litter

(Bishop et al., 2008). The phytophagous species,

which ingest organic material with more readily

degradable C, had greater assimilation capacities,

which resulted in lesser carbon content in cast

(Edwards and Bohlen, 1996). The feed quality such

as C/N ration, fibre content, and other secondary

metabolite are also reported to influence the C

states of castings (Mangold, 1951; Buck et al.,

1999).

In the earthworm castings, organic carbon

accumulates in different soil size fractions

(Guggenberger et al., 1996). However, distribution

pattern of C with all size fraction vary with soil

organic matter which they ingest and species of

earthworm. Organic matter associated with soil of

different size fraction (sand, silt and clay)

significantly exhibited different properties in terms

of organic matter turnover (Tiessen and Stewart,

1983; Anderson and Paul, 1984). Sand-associated

organic matter is important in short-term turnover,

clay-bound organic matter dominates the medium-

term turnover, whereas silt-bound organic matter

take part in long-term turnover. Therefore, particle-

size separation aids in estimating labile and passive

organic carbon fractions (Christensen, 1992).

In general, the clay bound C content seem to

be higher in the casts (Scullion and Malik, 2000;

Marhan et al., 2007). The organic matter bound to

clay is known to be stable and the formation of pool

of clay bound carbon is slow (Hassink and

Whitmore, 1997). Earthworms may enhance the

grinding and mixing organic matter with mineral

soil particles of the clay fraction thereby increasing

stabilized organic matter in soil (Oades, 1988). This

shows that clay bound organic matter is influenced

by gut transit. The C gains also high in fine-sand

sized fraction, almost twice as much C in the casts

as in the surrounding soil reported by Zhang et al.

(2003). Earthworm casts exhibited the lowest C

concentration in the sand-sized separates, but

compared with the sand-sized separates of the

surrounding soil they were much less depleted in

organic C. After gut passage a pronounced

depletion of C associated with sand and an

enrichment of silt- and especially clay-bound C is

reported. Hence, earthworm activity resulted in a

pronounced redistribution of C within size

separates (Guggenberger et al., 1996). In the

endogeic earthworm, Millsonia anomala about 25%

of C content with coarse fractions reduced during

gut transit, while C in the finer fractions tended to

increase. Similar changes were also encountered

with another species Polypheretima elongata in a

Martiniquan vertisol (Lavelle and Spain, 2003).

20

2.3.2. Macronutrients (primary)

Nitrogen – Nitrogen is critical to all the life

forms and is an important constituent of many

biologically-active compounds. It plays a

significant role in all the major category of

biological function in both animal and plant

(Lavelle and Spain, 2003). In terrestrial ecosystem

earthworms significantly influence the nitrogen

through modification of the physical, chemical and

biological properties of soil (Lee, 1985; Edwards

and Bohlen, 1996). In addition, earthworms

contribute to the mineral nitrogen pool of soil

directly due to excretion of nitrogenous compounds

in their urine and mucus (Whalen et al., 2000).

Castings they produce are reported to contain

higher total and extractable nutrients than the bulk

soil (Edwards and Bohlen, 1996), but the nitrogen

content of castings produced under field condition

was not always richer than bulk soil. The total

nitrogen content of casts is related to earthworm

diet and so may be similar to or greater than the

values reported for bulk soil, depending on nitrogen

in organic substrates ingested by earthworms (Syers

et al., 1979; Scheu, 1987; Buck et al., 1999;

Perreault et al., 2007; Kizilkaya, 2008). Flegel and

Schrader (2000) observed that the total nitrogen of

castings produced by Dendrobaena octaedra was

about twofold higher with dandelion as food,

compared to those fed with larch. In addition,

earthworm feeding habit also significantly

influenced its nitrogen value. For instance, the

castings of Octolasian tyrtaeum were of higher

mineral nitrogen with dry vetch as feed in

comparison to green vetch (Parkin and Berry,

1994).

Earthworm‘s castings are widely reported to

contain elevated amount of mineral nitrogen,

especially for the ammonium content in relation to

surrounding soil (Parkin and Berry, 1994; Decaëns

et al., 1999a; Whalen et al., 2000; Aira et al., 2003;

Bityutskii et al., 2012; Clause et al., 2013). High

mineral nitrogen content of fresh castings is

probably due to their preferential feeding of

nitrogen rich substrates and high rate of

mineralization of organic matter during gut transit

through their ―priming effect‖ (Lavelle et al.,

1995). Microorganisms ingested along with the soil

are stimulated by earthworm inner living conditions

and increase the mineralization of organic matter in

the earthworm gut (Lavelle et al., 1995; Aira et al.,

2003; Chapuis-Lardy et al., 2010), and this

continues for several hours in fresh casts (Barois

and Lavelle, 1986). In addition, the low

assimilation efficiencies of earthworms and urine in

the hind part of the digestive tract also could be the

reason for high ammonia content of vermicast

(Syers and Springett, 1983; Anderson et al., 1983;

Parkin and Berry, 1994; Aira et al., 2003).

In fresh castings, about 96% of mineral

nitrogen was present as ammonium (Parle, 1963). A

fraction (6%) of the non-plant available N ingested

by Allolobophora caliginosa (Savigny) was

excreted in plant available form (Syers et al., 1979).

The ammonium content of castings progressively

decreases with age (Decaens et al., 1999a;

Chevallier et al., 2006). The reason for the decrease

may be due to the NO3- production in fresh casts as

a result of nitrification (Scheu, 1987; Lavelle and

Martin, 1992). As the days progressed, the NO3-

content of vermicast also largely decreased

(Decaëns et al., 1999a), obviously due to root up-

take, immobilization in the soil microbial biomass,

denitrification processes or losses by leaching

(Syers et al., 1979; Elliott et al., 1990; Lavelle and

Martin, 1992).

Higher mineral nitrogen content and water-

filled pore space of vermicast and anaerobic

condition of the earthworm gut lead to increase in

the denitrification rate in the fresh cast (Elliott et

al., 1990; Knight et al., 1992; Granli and Bøckman,

1994; Smith et al., 1998), which is much higher

than the adjacent soil (Svensson et al., 1986; Elliot

et al., 1990; Parkin and Berry, 1994). Thus, gaseous

losses of nitrogen from vermicast continuous till the

production of nitrate ceased by intense nitrification.

Kharin and Kurakov (2009) observed about 1.5

21

times higher denitrification rate in fresh castings of

Aporrectodea caliginosa than that in the soil, and it

reduced by 30% and became closer to the activity

of denitrification in the soil after 12 days. In the

case of castings produced by Lumbricus terrestris,

about 2.5 times higher N2O emission is reported

than surrounding soil (Svensson et al., 1986).

Denitrification in cast may vary with different feed.

Parkin and Berry (1994) observed that castings

from worms allowed to feed on hairy vetch showed

higher denitrification rates than the castings

produced by earthworms which were manure fed or

not supplemented with organic material. The

denitrification rates of Octolasian tyrtaeum and

Aporrectodea tuberculata casts from vetch were

significantly higher than those from horse manure

or only soil. A few studies have shown that

earthworms make a substantial contribution to total

N2O emitted (30–56%) from the soils they inhabit

(Knight et al., 1992; Karsten and Drake, 1997;

Matthies et al., 1999; Borken et al., 2000).

Phosphorus – Phosphorus is one of a major

elements required by all the life form. It is an

essential component of ATP, ADP,

phosphoproteins, phospholipids of nucleic acids

and coenzymes. Phosphorus deficiencies lead to

suppression of many biological activities, so that

phosphalic fertilizers are widely used in agricultural

systems to improve the productivity of crops.

Phosphorus is one of the element which does not

have gaseous phase in its biogeochemical cycle.

Therefore phosphorus is largely cycled in soil by (i)

organic matter decomposition‚ (ii) turnover of the

microbial biomass‚ (iii) weathering of the parent

material and‚ (iv) in agro-ecosystems‚ as fertilizers

(Lavelle and Spain, 2003). In soil, the phosphorus

exists in different forms and concentrations.

However, most of phosphorus are associated with

aluminum‚ iron, calcium, manganese and other

carbonates, and became unavailable to plants. In

general, the availability of phosphorus in soil

mainly relies on pH and mineral composition of

soil, and few forms of phosphorus such as

dihydrogen phosphate and hydrogen phosphate that

can be easily utilized by plants (Lavelle and Spain,

2003; Chapuis-Lardy et al., 2009).

Earthworms are one of the important soil

organisms that can influence the availability of

phosphorus in soil. Through ingestion of particles

rich in P, the earthworms accumulate P in the casts.

They modify the proportion and stability of

different forms of P in castings (Brossard et al.,

1996; Kuczak et al., 2006) of species from tropical

and temperate regions (Sharpley and Syers, 1976;

Barois et al., 1987; Lavelle and Martin, 1992;

Lavelle et al., 1992; López-Hernández et al., 1993;

Scheu, 1987). Higher concentration of P in castings

may be probably due to (i) higher pH of the

intestinal gut contents (6.8, 6.0 and 4.6 for the

anterior and posterior parts and soil, respectively)

(Barois and Lavelle, 1986); (ii) a huge amounts of

mucus in the earthworm gut which is a mixture of a

glycoprotein, amino acids and sugars (Lavelle et

at., 1983; Martin et al., 1987) can inhibit and

compete for orthophosphate sorbing places by

carboxyl groups of carbohydrate compounds they

contain (López-Hernández et al., 1993), and (iii)

production of organic acids during digestion due to

increase in microbial activity which may compete

for P-sorbing sites and increase P solubility (Earl et

al., 1979; López-Hernández et al., 1993).

Moreover, enhanced phosphatase and microbial

activities in the biogenic structure of earthworm

and its surrounding increases availability of P

(organic and inorganic) (Sharpley and Syers, 1976;

Mulongoy and Bedoret, 1989; Edwards, 1998;

Richardson and Simpson, 2011).

The impact of earthworms on the availability

of P may be varying with different species, because

digestive abilities and gut microorganisms have

been reported to differ among species (Lattaud et

al., 1998). The casting of an epi-endogeic

earthworm, Lumbricus rubellus has been reported

to produce casts of higher exchangeable P, then

Lumbricus terrestris, an anecic species (Suárez et

al., 2003). The surface castings of Allolobophora

caliginosa is reported to contain about fourfold

22

more plant-available inorganic P and twice as much

plant-available organic P as the underling soil

(Sharpley and Syers, 1977), and those of

Pontoscolex corethrurus showed 2.7-fold increases

in water-soluble P after gut passage (López-

Hernández et al., 1993). However, increment in

available P was maintained for only few days, and

it decreased rapidly with age. Reduction in

available P was reported on fourth day and three

weeks after castings deposited by Pontoscolex

corethrurus (Lopez-Hernandez et al., 1993) and

Lumbricus terrestris, respectively (Basker et al.,

1993). The availability of high carbon and nitrogen

content in fresh cast increases the phosphorus

demand and subsequently immobilization P by

microorganisms may probably be the reason for P

decline.

Potassium – All the living organisms require

potassium, usually in relatively large amounts as

that of other major nutrients. It plays major role in

regulating many physiological and biochemical

processes in plants, such as pH stabilization,

osmoregulation, membrane transport and enzyme

activation and; thereby it influences photosynthesis,

protein synthesis and many other cellular and

physiological processes (Marschner, 1995). In soil

system, potassium is largely cycled by weathering

of primary minerals such as feldspars, micas, etc.

and mineralization of organic matter (Lavelle and

Spain, 2003). There are plenty of evidences on the

favorable effect of earthworms on soil potassium

status, through their borrowing and castings

activities. Many have reported a nominal increase

in total potassium and a significant increment in its

soluble form in vermicast compared to earthworm

feed or surrounding soil. However, the status of

potassium in castings may vary with different

earthworm species and feed they ingest.

Total potassium content in casting of Eisenia

fetida from cow dung, grass, aquatic weeds and

municipal solid waste increased to the extent of

28% compared to the raw feed (Pramanik et al.,

2007). Increase in potassium content is also

reported with castings produced from agro-

residues, kitchen waste, industrial and institutional

wastes including textile industry fibres and sludge

by the same earthworm species (Garg et al.,

2006a); whereas, castings were of lower potassium

content when they were fed with coffee pulp waste

(Orozco et al., 1996). However, castings from the

same coffee pulp waste showed higher potassium

content when it was produced by Eudrilus eugeniae

and Perionyx ceylanesis (Raphael and

Velmourougane, 2011). Increase in total potassium

content has also been reported with castings of

Eisenia fetida, Perionyx excavatus and Perionyx

sansibaricus with sewage sludge and anaerobic

digestate as feed, respectively (Delgado et al.,

1995; Rajpal et al., 2014). While casting of Eisenia

foetida and Eisenia andrei from excreta of different

livestock (cow, buffalo, horse, donkey, sheep, goat

and camel) and industrial sludges (paper mill, dairy

and leather industries), had lower potassium content

than the initial substrate (Elvira et al., 1998; Garg et

al., 2006b; Ravindran et al., 2008). These

differences in response of earthworms depends on

the chemical nature of the substrate and difference

in digestive ability of earthworm species.

In general, the increase in total potassium

content in castings probably due to ‗concentrative

effect‘, in which the loss of organic matter as CO2,

N2 or N2O during the gut transit, concentrated the

potassium content in ejected castings. Few studies

have also reported lower potassium in castings,

which might be caused by excess potassium uptake

by worms or leaching of soluble elements by excess

water that drained through the feed and vermicast

(Garg et al., 2006b; Ravindran et al., 2008;

Tajbakhsh et al., 2008). Leachate collected during

vermicomposting process is reported to contain

potassium in a significantly high quantity (Benitez

et al., 1999).

Many have reported significantly higher

exchangeable potassium content in castings in

comparison to the raw substrate or adjacent soil.

Castings produced in field had shown two to

23

threefold higher exchangeable K than the

surrounding soil (Tiwari et al., 1989; Bezborodov

and Khalbayeva, 1990; Hullegalle and Ezumah,

1991; Jouquet et al., 2008; Pommeresche et al.,

2009), however, it may vary with soil type and

species of worm which produced the castings

(Basker et al., 1994). For instance, in sandy soil,

the castings produced by Lumbricus rubellus had

higher K content than the Allolobophora

caliginosa; whereas, the castings produced in the

silty-clay soil had lower K in compared with

surrounding soil (Basker et al., 1994). Clause et al.

(2013) reported K content was not different

between soil and castings of Aporrectodea rosea

and Allolobophora chlorotica from Rendzic

Leptosol soil, but there was an increase in K

content with Lumbricus terrestris castings in the

Histosol. Laboratory based studies on Lampito

mauritii and Eisenia fetida with cow dung as feed,

showed 9.8–13.5% increase in exchangeable

potassium content in vermicast (Suthar, 2008).

Suthar (2007) reported that the exchangeable

potassium content in castings of Perionyx

sansibaricus had increased to the extent of about

20%, with guar gum industrial wastes. Similarly,

the castings of Perionyx excavatus and Perionyx

sansibaricus from cattle waste solids had twice the

concentration of exchangeable potassium than the

initial substrate (Suthar, 2009). Increase of

exchangeable potassium content in the castings

may be due to the release of K from the non-

exchangeable potassium pool in soil and organic

material during the gut transit (Basker et al., 1992,

1993) through, (i) secretion of acids by gut

associated microorganisms, which solubilize the

insoluble potassium in feed, and (ii) the enhanced

microbial activity in earthworm gut mineralizes the

organic bond potassium in their feed. However, the

release of K from the non-exchangeable pool is a

complex phenomenon which is controlled partly by

the level of exchangeable K in the ingested feed.

Higher exchangeable K in feed can inhibit the

release of further exchangeable K from non-

exchangeable pool (Feigenbaum et al., 1981;

Basker et al., 1994).

2.3.3. Macronutrients (secondary)

Calcium – Calcium is an essential element of

the cell walls of higher plants where it occurs along

with pectin. It plays major role in various

physiological processes in plants. In soil, calcium

strongly influences the pH status, and structural

stability of soil aggregates and so on (Lavelle and

Spain, 2003). Earthworms significantly influence

the soil Ca content by their castings activity. Many

species of earthworms produce calcite granules in

specialized glands called as calciferous gland,

which are three pairs of swellings away from the

oesophagus on the 10-12 segments (Canti, 1998;

Canti and Piearce, 2003; Clause et al., 2013;

Versteegh et al., 2014). These calcite granules are

produced by fixation of CO2 with Ca from their diet

under the catalytic reaction of carbonic anhydrase

present in calciferous glands, and these granules are

ejected along with their castings (Padmavathiamma

et al., 2008). Major portion of these granules

consists of calcite, and a small quantity of

amorphous calcium carbonate, aragonite and

vaterite also present in it (Gago-Duport et al., 2008;

Lee et al., 2008; Fraser et al., 2011; Brinza et al.,

2013). The production of calcite granules is likely

related to functions of regulating pH and the

concentration of CO2 in body fluids, and regulating

Ca2+

and other potentially toxic cations in their feed

(Robertson, 1936; Crang et al., 1968; Piearce,

1972; Bal, 1977; Becze- Deák et al., 1997).

The calcite granules production varies

significantly with different earthworm species and

the feed they ingest. Lumbricus terrestris is a major

calcite producing species in temperate soils.

Production rates are between 0.8 to 2.9 mg worm-1

day-1

(Canti, 2007; Lambkin et al., 2011; Versteegh

et al., 2013). Mršić (1997) and Udovic et al. (2007)

reported that Eisenia fetida, an epigeic earthworm

have active calciferous glands, while endogeic

earthworm, Octolasion tyrtaeum possess

calciferous glands that are intermediate in

complexity and activity. Increase in calcite granules

with elevated temperatures and atmospheric CO2

24

level, were also reported in few studies (Kühle,

1980; Versteegh et al., 2014). These granules are

very stable in nature, which are expected to survive

in soils for more than 300,000 years (Lambkin et

al., 2011; Versteegh et al., 2014).

In general, the vermicast are rich in calcium

content due to their calciferous glands activity and

higher mineralization capacity. Tajbakhsh et al.

(2008) reported about two to threefold increase of

calcium content in the castings of Eisenia foetida

and Eisenia andrei than the initial substrates of

compost produced from spent mushroom. The

castings of Eisenia fetida from sewage sludge,

coffee pulp waste and mixture of solid textile mill

sludge and poultry droppings also had higher Ca

content than its initial feeds (Delgado et al., 1995;

Orozco et al., 1996; Garg and Kaushik, 2005).

However, Elvira et al. (1998) and Kaushik and

Garg (2003) have reported a decrease in calcium

content in castings after processing of sludge of

paper pulp-mill, and solid textile mill with cow

dung by Eisenia andrei and Eisenia foetida,

respectively. The differences in the results may be

due to leaching of calcium by the excess water that

passed through the feed mixtures.

Calcium assimilation in the castings has

reported to be influenced by the soil type and the

concentration of calcium it contains. Castings

produced by Hyperiodrilus africanus had shown

twice the concentration exchangeable Ca than the A

horizons, and had about three times higher

exchangeable Ca than the B horizons in Lixisol and

Luvisol soils (Oyedele et al., 2006). Similarly, the

castings of Aporrectode caliginosa generated in

loam soil also showed more than two and threefold

higher Ca content compared to the soil of Ap and

Bt horizons, respectively (Schrader and Zhang,

1997). However, castings of same species had

lower Ca content than the P horizon of the clay soil.

Although, in the clay soil, Lumbricus terrestris

produced castings from the P horizon contained

lower exchangeable Ca than the soil in which they

produced, it was about double quantity of Ca found

in the castings of Ap horizon (Schrader and Zhang,

1997). While the Ca content of castings produced

by the Lumbricus terrestris, Allolobophora

chlorotica and Aporrectodea rosea in the Rendosol,

Luvisol and Histosol type soil were not

significantly different from their surrounding soil

(Clause et al., 2013). This could be attributed to the

absence or the very high content of CaCO3 in the

Luvisol and Rendosol soils, respectively which

might have masked impact of earthworms. The

availability of Ca is very scare in the few soil

systems like Apalachicola national forest, in which

the earthworm species Diplocardia mississippiensis

and Arctiostrotus sp. are believed to ingest fungi to

derive the oxalates bound calcium they contained

(Spiers et al., 1986; Lachnicht and Hendrix, 2001).

Ca present in the castings has various impact

on soil: (i) Ca2+

cation improve soil structure by

bridging with clay and soil organic matter, and

prevent the clay dispersion and the associated

disruption of aggregates by replacing Na+ and Mg

2+

in clay and aggregates, thereby enhancing the

stability of aggregates (Armstrong and Tanton,

1992; Zhang and Norton, 2002; Bronick and Lal,

2005), (ii) Ca preserve the flocculated structure of

the clay in soil surface aggregates by neutralizing

acids produced by fungi, microbes and roots

(Bullinger-Weber et al., 2007), (iii) Ca present in

the castings neutralize the acid soil by acting as a

buffering agent (Sanyal, 1991), (iv) calcium

influence the availability and solubility of other

plant nutrients present in the castings (Edwards and

Lofty, 1977), and (v) buffering capacity of Ca

present in the castings is also reported to enhance

the nitrifying bacterial populations in the

drilosphere (Parkin and Berry, 1999).

Magnesium – It plays important role in

regulating intra-cellular osmotic potential, protein

synthesis and it is an essential component in

chlorophyll synthesis. Magnesium is also

responsible for activating a wide range of enzymes

and, has a structural role in stabilizing the cell wall

(Marschner, 1995). Earthworm castings have been

25

found to contain elevated amounts of Mg relative to

adjacent soil (Tiwari et al., 1989; Parkin and Berry,

1994). The Mg content of castings produced by

Eudrilus eugeniae had increased to the extent of

40% compared to the initial substrates of rice straw

and rice husk amended with different proportion of

cow dung. Moreover, in these castings, Mg content

increased with increasing proportion of CD with

these feed (Shak et al., 2014). Similar increase in

Mg were also observed with castings of Eudrilus

eugeniae and Perionyx ceylanensis with mixture of

coffee pulp, cow dung and farm yard manure as

feed (Raphael and Velmourougane, 2011).

Asawalam (2006) reported about twice the

concentration of Mg in the castings than its

surrounding soil. Prakash and Karmegam (2010)

also reported about twofold increases in Mg content

of casings produced by Perionyx ceylanensis with

pressmud as feed. Castings of Eisenia sp. showed

20% increase in Mg content with apple pomace

waste (Hanc and Chadimova, 2014). While Eisenia

foetida and Eisenia andrei castings had about

threefold higher concentration of Mg with spent

mushroom compost (Tajbakhsh et al., 2008).

Pattnaik and Reddy (2010) also reported about

threefold increase in Mg content of castings

produced by Eudrilus eugeniae, Eisenia fetida and

Perionyx excavatus with vegetable market waste

and floral waste. The Mg content of castings also

reported to vary with different type of soil and its

layer (Basker et al., 1993; Pattnaik and Reddy,

2010). Suthar (2010) postulated that higher activity

of fungal and microalgae in fresh castings attributed

to the increase in concentration of Mg in castings.

Loss of organic matter during the gut transit also

may be leading to higher concentration of Mg in

castings (Wani et al., 2013).

Sulfur – Sulfur plays vital roles in a number

of metabolic pathways. It is a component of the

essential amino acids cystine‚ cysteine and

methionine and thus forms a structural part of tissue

proteins and enzymes important in photosynthesis

and nitrogen fixation. In most soils‚ sulfur occurs

largely in organic form and often associated with

the organic matter of soil and decaying plant

residues. This organic bound sulfur gets

mineralized to sulfate by the soil biota which can be

easily taken up by plants (Lavelle and Spain, 2003).

Among the different soil biota, earthworms

significantly influence the availability of sulfur in

the soil. Castings produced by them are reported to

contain higher amount of available sulfur than the

feed they ingest. Prakash and Karmegam (2010)

reported about twice the concentration of sulfate in

the castings of Perionyx ceylanensis with mixture

of pressmud and cow dung as feed. While sulfate

content of castings produced by Dravida

assamensis were sevenfold higher than the

surrounding soil (Ganeshamurthy et al., 1998).

Studies of Singh and Suthar (2012) also reported

higher sulfate content in castings of Eisenia fetida

with cow dung amended pharmaceutical industrial

waste, in which increasing proportion of cow dung

from 0 to 100% in initial feed mixture increased the

sulfate content about 50 to 270%, respectively in

the vermicast. However, the castings of similar

species with mixture of aerobic and anaerobic

biological sludges as feed showed lower sulfate

content than its initial feed mixture (Masciandaro et

al., 2000). Sangwan et al. (2010) reported no

difference between the vermicast generated from

cow dung and filter cake mixed with horse dung by

using Eisenia fetida. Increasing concentration of

available sulfur in the castings is probably due to

the mineralization of organic bound sulfur by the

enhanced microbial and enzyme activity in the

earthworm gut (Ganeshamurthy et al., 1998).

Leaching or gaseous loss of sulfur during the

bioconversion process might be the cause of sulfate

reduction observed in few studies.

2.3.4. Micronutrients

Micronutrients such as boron, chloride,

copper, iron, manganese, molybdenum, zinc and

nickel are important for all the plants (Brady and

Weil, 1999). Few other nutrients under this

category (i.e. sodium, cobalt, aluminum, etc.) are

considered to be essential for the plants growth,

26

however, certain species require these nutrients

only under specific environmental conditions. For

instance, plants that use C4 pathway of

photosynthesis and crassulacean acid metabolism

(CAM) require sodium (Welch and Shuman, 1995).

Leguminous plants require cobalt for symbiotic

fixation of nitrogen by Rhizobium bacteria in their

root nodules (O'hara et al., 1988). Availability and

mobility of these micronutrients in the soil is

largely influenced by earthworms‘ burrowing and

feeding activities. Castings they produced have

been reported to contain elevated amount of

micronutrients and which is largely assessed by the

nutrient levels of feed they ingest (Table 3). For

instance, castings of Aporrectodea caliginosa

showed higher availability of Fe, Zn and Mn in the

dried rye straw (Cecale cereale L.) treated soil than

those of clover aboveground parts (Trifolium

pratense L.) treated one (Bityutskii et al., 2012).

The majority of studies have shown that

increase in concentration of micronutrients in

vermicast in comparison to their surrounding soil or

feed they ingest (Ireland, 1975; Kizilkaya, 2004;

Wen et al., 2004, 2006; Asawalam and Johnson,

2007; Dandan et al., 2007; Udovic and Lestan,

2007; Udovic et al., 2007; Karmegam and Daniel,

2009). An elevated concentration of molybdenum

was recorded in castings of Aporrectodea

caliginosa which result in enhanced nitrogen

assimilation and fixation activities in maize

seedling through activation of Mo-depending

enzymes nitrate reductase and nitogenase,

respectively (Tomati et al., 1996). Lukkari et al.

(2006) also reported higher extractable metals in

the castings of Aporrectodea caliginosa tuberculata

than bulk soil. Oyedele et al. (2006) and Bartz et al.

(2010) also reported increase in Fe availability in

soil after processed by Pontoscolex corethrurus.

Increase in Fe content of castings attributed to

increase in amorphous form of Fe to available Fe

during the gut transit (Bartz et al., 2010). Oyedele

et al. (2006) postulated that higher contents of

extractable Fe and Al in casts are probably due to

transformation of crystalline fractions ingested soil

particles by the gizzard‘s crushing and mixing

activities. In addition, reduction of Mn and Fe in

the soils is facilitated by earthworm‘s anaerobic gut

passage due to lower redox potential of these

elements (Duarte et al., 2012).

The Fe, Mn and Zn content of Eisenia fetida

castings from municipal solid waste had increased

to the extent of 15, 32 and 15%, respectively

(Suthar et al., 2014). Bityutskii and Kaidun (2008)

have reported about five to tenfold increase in

water-soluble Fe and Mn in the fresh casting of

Eisenia fetida, Aporrectodea caliginosa, and

Lumbricus terrestris, and the solubility of these

nutrients increased further in 9 days older castings.

Among these earthworm species, Eisenia fetida had

showed lowest nutrients content in both fresh and

old castings. In sewage sludge amended soil,

Lumbricus terrestris produced castings of about

two and sevenfold higher Zn and Cu content than

its surrounding soil (Kızılkaya, 2004). Similarly,

castings of Pontoscolex corethrurus in the soil

contaminated with Pb mining activities had shown

elevated amount of Fe, Mn and Al oxides than its

surrounding soil (Duarte et al., 2012). Increase in

Fe, Z, Mn and Cu have also been reported with

vermicast from manure amended sludges from food

industries, dairy, paper mill, fly ash and winery

industries (Elvira et al.,1998; Nogales et al., 2005;

Venkatesh and Eevera, 2008; Yadav and Garg,

2011). Increase in concentration of micronutrients

in the vermicast could be due to (i) breakdown of

organic matter by the stimulated enzymes and

microbial activities in the earthworm gut, and

release of organically bound nutrients it contains

(Rada et al., 1996), and (ii) production of soluble

organics such as humic acids and fulvic acid during

the gut passage elevates the micronutrients

availability by the formation of water-soluble

complexes (Halim et al., 2003; Evangelou et al.,

2004).

Few studies have also reported lower

concentration of micronutrients in vermicast

compared to the initial feed or soil. For example,

27

Table 3. Some of the micronutrient content of castings produced by the different earthworm species from the different organic wastes.

Substrates Earthworm species Zn mg kg

-1 Cu mg kg-1 Fe mg kg

-1 Mn mg kg-1


Cattle dung Eisenia fetida 11.5 7.55 9.0 3 2.04 590.0 498.4 38.0 25.0 Bhat et al., 2013

Cattle dung Eisenia andrei 155.0 108.0 36.0 34.0 9600 6100 240.0 198.0 Elvira et al., 1998

Cow dung Eisenia fetida 155.0 117.3 - - 1368 1261 569.3 546.7 Suthar et al., 2014

Cow dung Eisenia fetida 193.0 145.0 52.6 32.4 2280 1810 - - Yadav and Garg, 2011

Dairy sludge + cattle dung Eisenia andrei 198.0 198.0 43.0 39.0 9300 7400 218.0 298.0 Elvira et al., 1998

Distillery sludge + cow dung Eisenia fetida 246.0 368.2 30.5 41.1 377.0 457.9 238.3 295.8 Suthar, 2008

Dyeing sludge Eisenia fetida 38.8 37.6 15.1 14.1 1600 1510 295.1 235.1 Bhat et al., 2013

Dyeing sludge + cattle dung Eisenia fetida 29.0 26.0 15.0 11.0 1456 1380 240.1 184.1 Bhat et al., 2013

Empty fruit bunch of oil palm Eudrilus euginae 10.6 2.82 9.59 2.18 9.29 1.62 18.75 16.8 Hayawin et al., 2010

Fly ash + cow dung Eudrilus eugeniae 14.0 8.60 11.8 1.20 64.9 27.5 29.6 12.9 Venkatesh and Eevera, 2007

Food industry sludge + poultry

droppings + cow dung Eisenia fetida 805.0 475.0 77.8 59.8 1400 1280 - - Yadav and Garg, 2011

Horse dung Eisenia fetida 917.0 870.0 387.0 153.0 25791 18630 1671 1121 Sangwan et al., 2008a

Itchgrass + cow dung Lampito mauritii 75.2 84.6 13.8 13.2 2300 2100 8300 9500 Karmegam and Daniel, 2009

Itchgrass + cow dung Perionyx ceylanensis 77.0 84.6 13.4 13.2 2400 2100 8700 9600 Karmegam and Daniel, 2009

Leaf litter of Indian mast tree +

cow dung Lampito mauritii 38.4 43.0 34.4 31.2 2100 1300 14800 16400 Karmegam and Daniel, 2009

Leaf litter of Indian mast tree +

cow dung Perionyx ceylanensis 36.3 43.0 31.5 31.0 1600 1400 15600 16300 Karmegam and Daniel, 2009

Municipal solid waste Eisenia fetida 236.7 201.3 - - 1603 1552 1043 1025 Suthar et al., 2014

Municipal solid waste + cow

dung Eisenia fetida 190.0 169.7 - - 1586 1445 891.3 834.0 Suthar et al., 2014

Paper mill sludge + cattle dung Eisenia andrei 108.0 110.0 34.0 31.0 7500 6900 190.0 180.0 Elvira et al., 1998

Pearl millet cobs + cow dung Lampito mauritii 46.0 63.0 27.0 28.8 2600 1900 800.0 1000 Karmegam and Daniel, 2009

Pearl millet cobs + cow dung Perionyx ceylanensis 52.6 63.0 27.4 28.8 2200 2000 900.0 1000 Karmegam and Daniel, 2009

Poultry droppings + cow dung Eisenia fetida 291.0 179.0 74.1 50.5 1398 1138 - - Yadav and Garg, 2011

Press mud + cow dung Perionyx ceylanensis 44.7 30.6 15.7 8.00 214.5 162.6 17.2 10.2 Prakash and Karmegam, 2010

Rice husk Eudrilus eugeniae - - 11.0 5.00 537.0 258.0 27.0 22.0 Shak et al., 2014

Rice husk + cow dung Eudrilus eugeniae - - 39.0 29.0 3980 2896 401.0 350.0 Shak et al., 2014

Rice straw Eudrilus eugeniae - - 10.0 6.00 2210 1372 280.0 175.0 Shak et al., 2014

28

Substrates Earthworm species Zn mg kg

-1 Cu mg kg-1 Fe mg kg

-1 Mn mg kg-1


Rice straw + cow dung Eudrilus eugeniae - - 45.0 33 4170 3566 499.0 450.0 Shak et al., 2014

Sewage Metaphire posthuma +

Lampito mauritii 518.4 542.0 35.4 42.6 529.0 563.0 397.4 477.2 Suthar et al., 2008

Sewage sludge Eisenia fetida 289.4 325.6 41.3 48.1 320.8 369.3 - - Suthar, 2010a

Sewage sludge + sugarcane trash Eisenia fetida 96.9 129.5 11.8 18.8 127.7 147.5 - - Suthar, 2010a

Sugar mill filter + horse dung Eisenia fetida 1554 1247 691.0 449 24716 22281 2569 1894 Sangwan et al., 2008a

Water hyacinth Eisenia fetida 243.8 184.2 46.3 45.9 17.1 10.3 585.0 430.1 Singh and Kalamdhad, 2013

Water hyacinth + cattle dung +

sawdust Eisenia fetida 218.1 186.3 46.8 35.6 6.60 6.90 600.0 476.0 Singh and Kalamdhad, 2013

Water hyacinth + cow dung Eudrilus eugeniae 268.0 210.0 96.9 52.1 79.1 71.0 162.3 126.1 Kumar et al., 2015

Winery industry waste Eisenia andrei 62.0 22.0 30.0 22.0 2497 623.0 53.0 8.00 Nogales et al., 2005

29

Suthar (2008a) reported the Zn, Fe, Mn and Cu in

distillery sludge amended with cow dung was

reduced to the extent of 39, 30, 38 and 42%,

respectively after processed by Eisenia fetida. Garg

and Kaushik (2005) and Gupta et al. (2005) also

reported a lower metal contents in the vermicast

compared to the initial feed of solid textile mill

sludge amended with poultry droppings and fly ash

amended with cow dung. Singh and Kalamdhad

(2013) have reported reduction in both total and

extractable form of Cu, Mn, and Fe in

vermicomposted water hyacinth with Eisenia

fetida. The reduction in these metals content was

also reported by other researchers (Jain et al.,

2004). The micronutrient availability in castings

probably determined by the properties of feed they

ingest, might be the reason for observed differential

response of earthworms reported in the previous

art. The reduction of metal nutrients may be

attributed to the tendency of earthworms to

accumulates metals in their tissues during the gut

passage (Hartenstein and Hartenstein, 1981; Graff,

1982; Gupta et al., 2005; Garg and Kaushik, 2005).

3. Effect of ageing on the properties of vermicast

In general, vermicast is rich in available form

of nutrients, beneficial microbial communities,

growth regulators such as auxins, gibberellins,

cytokinins in addition to enhanced physical

properties which can increase the nutrient and water

storage capacity, infiltration and aeration and

resistance to compaction and erosion in soil they

exist (Edwards, 2004). These beneficial properties

possess remarkable plant growth-promoting

potential on wide range of plants (Edwards, 2004;

Gajalakshmi and Abbasi, 2004). However, these

physical, chemical and biological attributes of the

vermicast are not stable in nature, and they change

with time (Hindell et al., 1997a,b; Decaëns et al.,

1999b; Parthasarathi and Ranganathan, 1999, 2000;

Decaëns, 2000; Tiunov and Scheu, 2000a,b;

Scullion et al., 2003; Aira et al., 2005, 2010;

Mariani et al., 2007; Kawaguchi et al., 2011).

Studies on vermicast from either soil or

blends of soil and phytomass have shown that fresh

castings are highly dispersible, which become more

stable during ageing (Marinissen and Dexter, 1990;

Hindell et al., 1997a,b; Piekarz and Lipiec, 2001).

Fresh castings produced by Lumbricus terrestris

and Lumbricus rubellus were up to 1.4 times more

dispersive than soil aggregates (Shipitalo and Protz,

1988), and those of Aporrectodea rosea,

Aporrectodea caliginosa and Aporrectodea

trapezoids were 19 times more dispersive than the

surrounding soil (Hindell et al., 1994). High level

of soluble carbohydrate in the castings was

connected to an increase in dispersion, which is

reported to be disintegrated and bonded to mineral

particles during drying and ageing (Hindell et al.,

1997a,b). Studies on endogeic species,

Aporrectodea caliginosa have shown that fungal

growth, drying and wetting cycle increases the

stability of castings during the ageing (Marinissen

and Dexter, 1990; Piekarz and Lipiec, 2001).

Shipitalo and Protz (1989) have found that ageing

and drying facilitates the bonding of plant and

microbial polysaccharides and other organic

compounds with clay (clay-polyvalent cation-

organic matter), thereby increases the stability of

vermicast. Close association of clay domains with

organic materials reduces the organic matter

decomposition they contain and adding to bond

longevity (Shipitalo and Protz, 1989; Guggenberger

et al., 1996). However, studies on epi-endogeic

earthworm, Lumbricus rubellus revealed that

microbial activity and presence of polysaccharide

did not have any influence on stability of castings

(Marinissen et al., 1996).

Impact of ageing on the bulk density of

casting was significantly varying with castings of

different earthworm species and the feed they

ingest. Decaëns (2000) reported that bulk density of

castings produced by Martiodrilus carimaguensis

was higher or equivalent to that of the surrounding

soil which rely on the organic matter it contains.

The bulk density of casts did not alter in the first

week after deposition, but then decreased

30

progressively to the extent of 29%. Decaëns (2000)

also found that regular shape and thin cortex at their

periphery of castings had largely disappeared

during the ageing. Old castings also contains cracks

and void space which were formed by degradation

of plant debris present in fresh castings and

ingestion of castings by other invertebrates.

Fresh vermicast are reported to contain

higher content of organic carbon than its

surrounding soil (Lee, 1985; Shipitalo and Protz,

1988; Daniel and Anderson, 1992; Barois et al.,

1993; Zhang and Schrader, 1993; Buck et al., 1999;

Bossuyt et al., 2005), whereas castings from purely

phytomass or manure have showed reduction in

carbon content than the feed they ingest (Aira and

Domínguez, 2009). Contradiction in the results is

due to the preferential ingestion of high organic

matter by earthworm in field, and however, C

content of castings compare with only the bulk soil.

During ageing, the organic carbon present in the

castings mineralized by microbial activity, leading

to reduction in the carbon pool in the vermicast

(McInerney and Bolger, 2000a). However, in the

studies on castings deposited in nature, a

continuous increase in C was observed during their

ageing (Jiménez and Decaëns, 2004). The reason

may be fixation of atmospheric CO2 by algae or

nitrification bacteria (autotrophic microorganisms);

colonization of casts by cast-dwelling macro-

invertebrates and accumulation of organic material

and/or the production of carbon-enriched feces by

earthworms fed on organic-rich food substrates

(Jiménez and Decaëns, 2004). The organic matter

in the ageing vermicast can be protected over

longer periods of time from further decomposition

and might then become available for the microflora

once these are broken down into small fragments

(Blanchart et al., 1997; Decaëns, 2000; Bossuyt et

al., 2005).

The vermicast are widely reported to contain

elevated amount of mineral nitrogen, especially for

the ammonium content relative to surrounding soil

(Parkin and Berry, 1994; Decaëns et al., 1999a;

Whalen et al., 2000; Aira et al., 2003; Bityutskii et

al., 2012; Clause et al., 2013). During ageing, the

casts were characterized by a high rate of

nitrification, which reduced the ammonium content,

and simultaneously increase nitrates. Kharin and

Kurakov (2009) have reported that the castings of

endogeic earthworm, Aporrectodea caliginosa

showed two to threefold increase in nitrate content

during the 12 days of ageing. Further ageing,

reduced the denitrification activity, and became

closer to their surrounding soil. The nitrogen

present in microbial biomass also greatly affect by

the ageing, thus in 45 days old castings of

Aporrectodea caliginosa it reduced to the extent of

33% (Aira et al., 2005). The activities of enzymes

such as dehydrogenase, β-glucosidase, cellulase,

invertase, amylase, protease, phosphatase of

castings were also reported to reduce during the

ageing (Aira et al., 2005; Kharin and Kurakov,

2009). The reduced enzyme activities in aged casts

were probably due to: (i) the reduced nutrient and

moisture contents, (ii) a decline or inactivation of

the microbial population, (iii) low stability of aged

earthworm casts, and (iv) degradation or

inactivation of enzymes during the ageing (Aira et

al., 2005).

Microbial diversity and its activity in castings

were strongly affected by age. Fresh castings are

reported to contain higher microbial biomass than

the feed they ingest (Parthasarathi and

Ranganathan, 1999; Tiunov and Scheu, 2000a,b).

Ageing have been showed predictable microbial

successions with castings of different species.

During ageing, the bacterial activity reduced due to

shifting from bacterial to fungal dominance within

the population (Scullion et al., 2003; Piekarz and

Lipiec, 2001; Aira et al., 2005). In the castings of

Lumbricus terrestris and Lumbricus rubellus, the

bacterial population largely increased in the first

four days and then there was rapid decline (Scullion

et al., 2003). Similarly, the castings of

Aporrectodea caliginosa also showed increase in

bacterial population in the initial days followed by

fungal dominance over the bacterial population

31

during the first week of ageing. Even though,

further ageing reduced the fungal population, it

remained four times more than that in the soil

during the 12 days ageing (Kharin and Kurakov,

2009). Castings deposited in the field are found to

be colonized by macro-invertebrates after 4 to 6

weeks of ageing. The density of invertebrates found

inside casts was reported to much lower than those

found in soil. The total count of individuals inside

the casts or in the underlying soil was not changed

during cast ageing. Moreover, only a small number

of specialized (i.e. small and mobile) species are

able to live inside vermicast (Decaëns et al.,

1999b).

4. Effect of vermicast on plant growth and soil

Vermicast contains all the essential macro

and micronutrients for plants growth, regardless of

different substrate used to generating vermicast.

The nutrient present in the vermicast are more

bioavailable (Gajalakshmi and Abbasi, 2008;

Edward et al., 2011), and the nutrients bound in

organic matter are release gradually through

mineralization of the organic matter, thus causing

lesser nutrient loss from the rhizosphere (Chaoui et

al., 2003). Vermicompost also improves the

physical properties of the soil, such as aeration,

water holding capacity and porosity which all have

direct impact on the plant productivity in vermicast

applied soil (Edwards, 2004). Additionally the

beneficial impact of vermicast is also attributed to

biologically active substances such as fulvic acids,

humic acids and phytohormones it contains.

Specifically, cytokinins have been reported in

vermicast (Zhang et al., 2014). It has also been

shown that humic acids derived from the vermicast

induce morphogenetic and biological changes

favorable to plants which are similar to those

produced by indole-3-acetic acid (Muscolo et al.,

1999). These bioactive substances are probably

produced due to the abundance of microbial

communities in the vermicompost, specifically the

actinomycetes and fungal species, which then

releases phytohormones in the soil (Frankenberger

and Arshad, 1995). Enhancement of all these

properties of soil with vermicast supplement are

reported to increase productivity of many plants

(Tomati et al., 1983, 1988, 1990, 1995; Abbasi and

Ramasamy, 1999; Atiyeh et al., 2002; Arancon et

al., 2003, 2004; Gajalakshmi and Abbasi, 2004;

Acevedo and Pire, 2004; Edwards, 2004; Sinha,

2009).

It is also postulated that vermicompost can

suppress a wide range of microbial diseases, insect

pests and plant parasitic nematodes. Availability of

adequate nutrients with vermicast application

enhances the ability of the plants to limit the

penetration, development, and/or reproduction of

invading pathogens (Graham and Webb, 1991),

which contributes to the development of resistance

to pathogens in the plants. Some of the nutrients are

also involved in the production of antimicrobial

compounds such as flavonoids and phenolics that

act against the plant pathogens (Graham and Webb,

1991; Hill et al., 1999). The humic acid content of

the vermicast is also likely to affect biochemical

processes in the plants and bacteria, resulting in

induction of resistance in plants to certain

phytopathogens (Sahni et al., 2008). Vermicast

application also enhances the activity of diverse

beneficial microbes in soil, which may act as

biocontrol agents to suppress plant pathogens

(Gunadi et al., 2002; Arancon et al., 2006). Prior

art shows that the vermicast significantly reduced

sporulation of the pathogen Phytophthora

cryptogea (Orlikowski, 1999), reducing the growth

of pathogenic fungi such as Botrytis cinerea,

Sclerotinia sclerotiorum, Corticium rolfsii,

Rhizoctonia solani and Fusarium oxysporum

(Nakasone et al., 1999), reduced infection of

Fusarium lycopersici (Szczech, 1999) and

Phytophthora nicotianae (Szczech and Smolinska,

2001) in tomato seedlings. Vermicast also reported

to be reducing the infestation of Heteropsylla

cubana (Biradar et al., 1998), Aproaerema

modicella (Ramesh, 2000), and many other insect

pests and mites in various plants.

32

Vermicast effect on density and water related

properties on the soil rely on castings age and

species that produce them. Aged castings are more

stable than its surrounding soil, whereas the fresh

castings are easily dispersible, in water, and thereby

effecting clogging of transmission pore and

impeding the infiltration in soil (Shipitalo and

Protz, 1988; Le Bayon and Binet, 1999). The aged

castings being stable get protected from detachment

by rainfall or surface flow, and as a consequence

increase infiltration. The shape, size and

compaction of castings also have differential

impact on soil properties. Castings produced by

‗compacting‘ species such as Pontoscolex

corethrurus and Millsonia anomala are reported to

impede the infiltration, increase the bulk density

and reduce the porosity of soil (Alegre et al., 1995;

Barros et al., 1996; Young et al., 1998; Chauvel et

al., 1999; Hallaire et al., 2000; Spain et al., 1992;

Derouard et al., 1997; Blanchart et al., 2004). In

contrast, larger size compacting type castings

produced by Amynthas khami are reported to

improve water infiltration by their large packing

voids between castings which are connected with

below-ground galleries (Jouquet et al., 2008);

whereas, the castings of decompacting type are

highly dispersible and impedes the infiltration rate

(Shipitalo and Protz, 1988; Le Bayon and Binet,

1999). The large size castings of compacting

species also create anaerobic condition inside the

castings, which reduces the rate of decomposition

of organic matter it contains (Blanchart et al.,

1993). In the case of smaller castings produced by

the decompacting species, the decomposition rate is

accelerated due to high microbial and enzyme

activity along with favorable physical condition for

the growth of aerobic microorganisms. Similarly, in

the fresh castings, the organic matter has fast

decomposition rate and it is stabilized within

microaggregates formed within the casts during the

ageing (Six et al., 2014). Thus, the casting of

different age of different species also has

differential impact on the soil carbon stock.

5. Conclusions

Vermicast produced by different earthworm

species has different physical, chemical, biological

characteristics, which rely on the habitat and

feeding behavior of earthworms that produce them.

Thus, the impact of castings of different earthworm

species on the soil and plant also vary significantly.

In general, castings of all the species have higher

plant available nutrient than their surrounding soil

or the feed they ingest. The presence of growth

regulators, beneficial microbes, and enhanced

enzymes activity associated with nutrient

mineralization are also reported in castings of many

species. Castings also improve the physical

properties of soil by reducing bulk density and

erosion, enhancing porosity, infiltration, water

holding capacity and so on. However, most of the

previous studies on vermicast in relation to soil

properties such infiltration, compaction, erosion,

soil carbon stock, structural changes and other

related aspects have been studied with anecic and

endogeic species. While, the castings of epigeics

has been extensively studied in relation to plant

growth. But not much focus has been paid to

understand the impact of castings of anecic and

endogeic species on seed germination, plant growth

or disease suppression, or vis-à-vis.

Moreover, the physical, chemical and

biological attributes of the vermicast are not stable

in nature, and they change with cast ageing.

Previous studies on the fate of vermicast over time,

have mainly focused on the stability of castings

deposited by anecic and endogeic earthworm

species and very few studies have been done on

epigeics. The castings used in these studies were

also produced from non-specific substrates in

nature or from blends of soil and phytomass.

Therefore, the castings may contain considerable

amount of soil particles, which is known to increase

the stability of these biogenic structure, slow down

the decomposition of organic matter, and also either

33

stabilize or bring about proteolysis of the

extracellular enzymes it contains. Hence, the

findings of these studies cannot be used as a model

to study the dynamics of the properties of castings

when it is produced by anthropogenically

controlled vermicomposting systems, which are

purely derived from the organic matter. It is

therefore necessary to conduct more studies on

these aspects to improve our understanding on the

interaction of these biogenic structures with agro-

ecosystems, so that the beneficial features of

vermicast can be utilized maximally.

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INGESTION OF SAND AND SOIL BY PHYTOPHAGOUS

EARTHWORM EUDRILUS EUGENIAE: A FINDING OF

RELEVANCE TO EARTHWORM ECOLOGY AS WELL AS

VERMITECHNOLOGY

Chapter

3

53

A paper based on this chapter has been published in

Archives of Agronomy and Soil Science, 60 (12), 1795 – 1804, 2014.

CChhaapptteerr 33

Ingestion of sand and soil by phytophagous earthworm

Eudrilus eugeniae: a finding of relevance to earthworm

ecology as well as vermitechnology

Abstract

Epigeic earthworms are phytophagous in habit and are believed to prefer organic matter, principally

phytomass, in various degrees of decay. To check whether epigeics will ingest soil/sand even when there is

luxury availability of phytomass, the present work was carried out. It was seen that not only soil but even

sand particles were ingested by epigeic (Eudrilus eugeniae) tested by us, even as there was availability of

phytomass and cow manure in abundance. But the consumption of sand and soil decreased as the days

progressed. Moreover, the ingestion of sand and soil with phytomass and cow dung did not show any

significant influence on growth and survival of earthworms. It did increase the vermicast mass due to the

presence of soil/sand as was confirmed by microphotography using polarizing microscope. The findings have

important implications in the design of vermireactors for maximizing their efficiency.

1. Introduction

Epigeics are humus feeding worms which

dwell at or very near the surface of the soil horizon.

Due to their preference for phytomass as food,

epigeics are also called phytophagous in contrast to

the anecics which are termed geophytophagous due

to their tendency to ingest soil together with

organic matter, and endogeics which are called

geophagous due to their preference for soil. It is

generally believed that periodic ingestion of

sand/soil is essential for even epigeics to keep their

gizzard muscles toned up. As gizzards play a

pivotal role in comminuting the material that the

earthworm ingest, it has been believed that hard-to-

break particulates as are contained in sand/soil

should be always a part of earthworm diet if they

have to remain healthy (Marhan and Scheu, 2005).

Although this belief is well-entrenched, there is no

existing report which establishes its veracity. This

is despite the fact that the role of epigeics in the

degradation of various types of leaf litter and other

organic wastes has been explored in detail (Lim et

al., 2012, 2014; Shak et al., 2014). Information is

also available on the attempts to use fly ash (Saxena

et al., 1998; Gupta et al., 2005), rock phosphate

(Wang et al., 2013), and sludges of textile mills

(Garg and Kaushik, 2005), tanneries (Hemalatha

and Meenabal, 2005), petrochemical industries

(Banu et al., 2005), pharmaceutical industries

(Majumdar et al., 2006), and distilleries (Suthar and

Singh, 2008) as epigeic feed after amending it with

animal manure or sewage sludge. But not much

attention has been paid towards determining

whether ingestion of soil/sand is essential for long-

term survival and fecundity of epigeics.

Eudrilus eugeniae is known to be a voracious

feeder of animal manure, especially cow dung

(Fernández-Gómez et al., 2013), but it’s acceptance

for phytomass is not as well- established. Indeed

except a few studies by Gajalakshmi, S.A. Abbasi

and co-workers (Gajalakshmi et al., 2002; Makhija

et al., 2011), no one has reported direct

54

vermicomposting of phytomass – without pre-

composting or subsequent blending with animal

manure – by E. eugeniae. On the other hand, by

employing the concept of high-rate

vermicomposting and associated technology

developed by Abbasi and co-workers (Gajalakshmi

et al., 2002, 2005; Abbasi et al., 2009, 2011;

Tauseef et al., 2013), several types of phytomass

have been recently vermicomposted using

E.eugeniae and other species in reactors which had

no sand/soil or any other bedding, and which were

fed unprocessed phytomass in the form of leaf

litter, weeds, vegetable waste etc. Also the reactors

could be run in pulse-fed mode for as long as one

wished, with the epigeic species placed in them not

only surviving but gaining in zoomass and

exhibiting good fecundity. Hence, in the context of

vermireactors design for attaining maximum

vermiconversion efficiency it has become important

to answer the question: Is presence of sand/soil

really essential? There is also a need to establish

whether E. eugeniae may ingest soil/sand if forced

to feed upon phytomass but may not do so when

fed with animal manure. Hence, we chose cow

manure, leaves of neem (Azadirachta indica) and of

ipomoea (Ipomoea carnea) as substrates for the

present study. Neem and ipomoea were chosen

because both are likely to be inimical to, rather than

preferred by, E. eugeniae as neem contains several

chemicals known to repel or kill invertebrates while

ipomoea contains toxic alkaloids. We also studied

the influence of different earthworm densities (25,

15 and 5 animals per kg of substrate) on feed

consumption, as crowding may induce a tendency

to forage beyond the phytomass layer and into the

sand/soil bedding. Of the epigeic species studied

most extensively vis-a-vis vermicomposting so far

– Eisenia fetida (Savigny), E. eugeniae (Kinberg)

and Perionyx excavatus (Perrier) (Sim and Wu,

2010) – E. eugeniae was chosen by us as it is the

most voracious of feeders among the three

(Gajalakshmi et al., 2002, 2005; Edwards, 2004;

Lim et al., 2011). In addition, epigeics may have

advantage over burrowing earthworms in cutaneous

absorption and dietary intake due to their surface-

dwelling mode of life (Suthar, 2014).

2. Materials and methods

Ipomoea and neem leaves were collected

from in and around the Pondicherry University

campus. The leaves were washed with water to

remove adhering material, then soaked for 48 hours

and lightly squeezed to soften them. Rectangular,

41 liter wooden boxes (30 cm high with surface

area of 35 x 39 cm) were used as vermireactors.

They were lined up with thick transparent plastic

sheets to prevent water loss, as also to prevent

earthworms from escaping or predators from

coming in.

Five sets of vermireactors were employed.

The first set consisted of reactors which were filled

with a layer of coarse sand to a height of 3 cm

followed by a 5 cm high layer of soil. Over it 1kg

dry weight equivalent of cow dung was placed

which has raised the reactor content further by

about 7 cm. Other reactors in this set consisted of

ipomoea or neem instead of cow dung. The heights

of the substrates in these reactors were 12 and 7.5

cm, respectively. The second set of reactors was

identical to the first set except that there were

without sand/soil. The third set comprised of the

control reactors which had only soil and sand

bedding but no substrate. Bunches of 25 adult

earthworms, weighing 0.9 - 1.5 g and length of 9 -

12 cm, randomly picked from their cow dung –

based culture, were introduced in each of the

reactors. The fourth and fifth sets of reactors were

identical with the first set except that the number of

earthworms incorporated therein was 15 and 5

separately.

All the reactors were operated at identical

ambient conditions with temperature of 29 ± 4°C.

Moisture was maintained by periodic sprinkling of

adequate quantities of water. The reactors were

operated continuously for 8 runs of 15 days each.

At the end of each run, vermicast in all the reactors

55

Table 1. Major and trace elements composition of plant leaves, cow dung, soil and sand used in this study

Parameters Neem leaf Ipomoea leaf Cow dung Soil Sand

Dry matter % 34.0±0.4 21.9±0.2 26.5±0.3 98.6±0.0 98.3±0.2

Organic carbon mg g-1

616.1±5.9 656.9±4.4 359.9±8.0 4.30±0.07 0.39±0.15

Nitrogen mg g-1

32.7±1.5 39.6±2.3 25.4±0.5 0.34±0.10 0.11±0.06

Phosphorus mg g-1

0.20±0.00 0.26±0.01 0.62±0.00 0.25±0.03 0.16±0.01

Potassium mg g-1

16.0±0.1 39.2±0.3 7.38±0.42 5.49±0.15 15.0±0.4

Sulphur mg g-1

6.58±0.05 6.89±0.34 7.78±0.42 0.07±0.06 0.16±0.05

Calcium mg g-1

16.5±0.2 11.2±0.3 4.91±0.23 3.51±0.06 12.8±0.7

Sodium mg g-1

0.56±0.03 0.73±0.05 3.94±0.09 2.45±0.02 11.4±0.6

Magnesium mg g-1

6.58±0.14 2.48±0.04 4.37±0.18 1.04±0.03 3.47±0.36

Copper mg kg-1

7.89±1.69 21.1±1. 6 25.1±4.4 – 33.3±28.9

Iron mg g-1

0.22±0.01 0.36±0.00 4.75±0.42 63.0±0.7 30.7±24.0

Manganese mg kg-1

44.8±1.6 85.2±4.8 274.0±10.9 0.03±0.02 200.0±173.5

Zinc mg kg-1

29.1±0.7 34.1±3.9 69.0±2.6 0.01±0.00 10.0±17.3

was harvested and the reactors were restarted with

the leftover substrate. The castings were carefully

separated from other particles by soft painting

brush and quantified. While disbanding the

reactors, the adult earthworms with which the

experiment was started, were washed, blotted dry

and weighed to record their zoomass before they

were put back in the respective reactors. Dead

earthworms, if any present, were replaced, while

starting the next run.

Total organic carbon content in the soil, sand

and plant material was measured by modified

dichromate redox method (Heanes, 1984). Total

nitrogen content was determined by modified

Kjeldahl method (Kandeler, 1993) using Kel Plus™

semi-automated digester and distillation units

(Pelican Equipments, Chennai, India). Total

phosphorus content of the plant material was

determined by ammonium molybdate –

hydroquinone method (AOAC, 2012). Potassium,

calcium and sodium were determined using an

Elico CL378 model flame photometer (Elico Ltd,

Hyderabad, India) after dry ashing the plant

material at 500°C for 4 hours and dissolution of the

ash in hydrochloric acid (Kalra, 1998). Magnesium,

boron, copper, iron, manganese, zinc and

molybdenum concentration in plant samples were

determined with thermo electron IRIS intrepid II

XSP DUO model inductively coupled plasma

atomic emission spectroscopy (ICP-AES, Thermo

Fisher, Waltham, MA, USA) as per procedure

given in AOAC (2012). The soil and sand samples

were analyzed for their mineral content by

Bruker™ S4-Pioneer model wavelength dispersive

X-ray fluorescence spectrophotometer (WD-XRF,

Bruker, Billerica, MA, USA). The samples were

ground to 100 µm particle size using ball mill. The

processing of emission spectra for identification of

elements and their peak area measurement was

carried out using SPECTRAplus® software, version

1.6 (Pioneer Hill Software, Poulsbo, WA USA).

The results are summarized in Table 1.

The presence of sand, soil and organic matter

in the vermicast was quantified by gravimetry.

Samples oven dried at 105 °C to a constant weight

were put in distilled water, and crushed to release

the soil/sand. The content was filtered through a

Whatman No. 42 filter paper. The residue over the

filter paper was kept in a muffle furnace at 550°C

for 4 hours to remove organic matter present in the

castings (John, 2004). To observe the internal

structure of casts, 1mm thin sections of the castings

were made, after impregnating them in araldite –

xylene mixture, with the help of Buehler

56

PetroThin® thin sectioning system (Buehler, Lake

Bluff, IL, USA). They were studied using a

polarizing microscope at low magnification (4x)

(FitzPatrick, 1993). The surface area of casting and

sand grains in the sections were measured with the

Leica® software, version 3.8.0 (Leica

Microsystems, Heerbrugg, Switzerland).

Differences in the significance of vermicast

production, earthworm mortality, zoomass gain and

sand and soil content in the vermicast between

different treatments were tested by repeated

analysis of variance. The cast size and grain

covered area in the vermicast sections were

analyzed with two-way ANOVA at the 99.9%

confidential level. SPSS 16 package (Softonic,

Barcelona, Spain) was used for all statistical

analysis.

3. Results and discussion

3.1. Vermicast output

There was significant difference (p<0.01) in

vermicast production between reactors with

different substrates, worm density and presence and

absence of sand/soil bedding (Figure1 and Table 2)

indicating the influence of these factors on

vermicast production. In general, the chemical

nature of the organic waste influences the

palatability of earthworms directly or indirectly,

which consequently affects the earthworms’

efficiency in processing a substrate. In the present

study, vermireactors with the presence of sand/soil

bedding, produced maximum amount of vermicast

in conjunction with cow dung, followed by neem

and ipomoea. This is consistent with the belief that

earthworms are attracted by most kinds of animal

dung (Lowe and Butt, 2005; Marhan and Scheu,

2005), due to partially decomposed nature and high

nitrogenous organic matter content of animal dung.

The quantity of cast generated with cow dung as

feed and with sand and soil bedding was 643±100,

701±70 and 1392±10 mg cast worm

-1 d

-1 with 25,

15 and 5 worms per kg dry weight of substrate,

respectively. The lowest cast production was seen

when the ipomoea was offered as a feed: 251±10,

308±70 and 423±80 mg cast worm-1

d-1

castings

with 25, 15 and 5 worms, respectively. Ipomoea is

reported to consist a variety of toxic alkaloids such

B1, B2, B3, C1 and N-methyi-trans-4-hydroxy-L-

proline (Asano et al., 1995; Molyneux et al., 1995;

Cholich et al., 2013). It is possible that the presence

Figure 1. Vermicast output, g worm -1

day-1

(mean ± SD) recorded in reactors with different treatments.

(‘w’ represent the worm density per kg substrate).

0

0.2

0.4

0.6

0.8

1

1.2

1.4

1.6

5w 15w 25w 5w 15w 25w 5w 15w 25w 25w 25w 25w 25w

Neem Cow dung Ipomoea Neem Cow dung

Ipomoea

With Bedding Without Bedding Bedding alone

Ve

rmic

ast

ou

tpu

t, g

w -1

d-1

Treatments

57

Table 2. Repeated analysis of variants and ANOVA table of F-values and the effects of substrate, bedding

and worm density on vermicast output, average zoomass changes, mortality, sand and soil entrapped in

castings, castings surface area and sand grains covered area of castings.

Repeated analysis of variants ANOVA

df Vermicast

output

Average

zoomass

changes

Mortality

Sand and

soil

entrapped

Casting

surface

area

Sand grains

covered

area

Substrate 2 63.18***

152.0***

16.27**

123.47***

105.31***

102.44***

Bedding 1 36.87**

7.58 n.s

215.4***

– 12.75***

–

Worm density 2 60.23***

9.76**

1.01n.s

1.27n.s

4.33* 3.50

*

Substrate x bedding 1 2.80 n.s

0.201 n.s

30.99***

– 37.41***

–

Substrate x worm density 4 3.12 n.s

6.38* 2.019

n.s 2.03

n.s 3.243

* 3.41

**

Bedding x substrate x

worm density 6 – – – – 17.86

*** –

*p<0.05,

**p<0.01,

***p<0.001,

n.s - not significant.

of these chemicals led to less intense feeding and

lower cast production in comparison to the other

substrates.

In the case of neem-based reactors, the

amount of cast production was higher than in

ipomoea but lower than in cow dung. In these

reactors 496±40, 622±40 and 1037±80 mg

cast

worm-1

d-1

were produced with 25, 15 and 5 worms,

respectively. Even though neem is known to have

nematicidal and insecticidal properties (Dai et al.,

2001; Nathan et al., 2005) it does not seem to have

a deleterious effect on earthworms. Earlier

Gajalakshmi and Abbasi (2004) and

Selvamuthukumaran and Neelanarayanan (2012)

have reported that earthworms fed upon the neem

leaf compost even more voraciously than they did

on the compost of mango leaf and ashoka tree leaf.

However, the presence of polyphenolic compounds

might have affected the earthworm in fresh

substrate. The antagonism between earthworms and

polyphenols/lignin content of their feed has been

reported in studies by Satchell and Low (1967),

Hendriksen (1990), Tian et al. (2000) and Ganesh

et al. (2009). These studies showed a negative

correlation between the palatability of leaf litter and

its total polyphenol content in both natural and

planted fallows.

The pattern of vermicast output from

different treatments is illustrated in Figure 1. The

results of cast generated per worm per day with

different earthworm densities showed decreasing

trend with increasing worm density. The maximum

vermicast generated was 1392±10, 1037±80 and

423±80 mg cast worm

-1 d

-1 in cow-manure, neem-

and ipomoea-based reactors, respectively, run with

5 earthworms. Minimum vermicast productions of

643±100, 496±40 and 251±10 mg cast worm

-1 d

-1

were observed in cow manure, neem and ipomoea,

respectively, in reactors with 25 earthworms. The

results showed remarkable differences between the

masses of vermicast generation in reactors which

consisted sand/soil and ones which did not.

Reactors without sand/soil and with 25 worms

yielded 316±20 and 310±40 mg worm

-1 d

-1 cast

with cow manure and neem, respectively. A

significant amount of sand and soil entrapped in the

castings may be the reason for the higher mass of

vermicast obtained from reactors with sand/soil

bedding compared to the ones without this bedding.

However, there was no significant difference in net

58

output of vermicast from reactors with and without

sand/soil. Very few castings were seen in ipomoea-

fed reactors without bedding, due to the 100%

mortality in all the three runs. In the case of control,

very few castings were observed. The casts were

less stable and easily disintegrated while

moistening the reactors. Hence, the castings

generated in the control reactors could not be

studied further.

3.2. Mortality

There was mortality of earthworms in the

first run in most of the reactors. The type of

substrate and the presence of sand/soil had a

significant effect on the survival of the earthworms

(p<0.01) (Table 2). The difference was more

pronounced in ipomoea-based reactors. Ipomoea

reactors with sand/soil showed 100% and ~40%

mortality in the first and second runs, respectively,

but had no mortality from then onwards. In contrast

in ipomoea reactors without sand/soil, 100%

mortality was observed consistently from the

beginning to third run. Hence, these reactors were

not operated further. In neem reactors with bedding,

a maximum of 84% mortality was seen in one of

the reactors with 25 earthworms and in the

remaining reactors there was no significant

mortality in the first run. In the case of reactors

without sand/soil, a maximum of 52% mortality

was recorded in the first run. However in reactors

with and without sand/soil, there was no mortality

from the second run onwards. In the case of cow

dung-fed reactors, there was no mortality in any of

the runs. The reactors with soil only as bedding and

without substrate did not show any mortality up to

third run, but from fourth run, the mortality rate

increased in these reactors. The reason may be that

in these reactors the earthworms, which are

phytophagous, were forced to feed exclusively on

sand/soil and could not survive on it for long.

3.3. Average zoomass change per animal

There was a significant influence (p<0.01) of

different substrates and worm densities on

earthworm zoomass, but there was no difference in

zoomass between reactors with and without

sand/soil (Table 2). In the first run, there was a

decrease in zoomass in most of the reactors with

neem and ipomoea, but from the second run

onwards there was a gain in zoomass in all the

reactors. With cow dung as feed, the increase in

zoomass was recorded in all the reactors from the

beginning to the eighth run. The results also show

that there was no significant difference in zoomass

change between reactors with 5 and 15 worms;

however, further increase in worm density to 25

showed a decline in zoomass gain in comparison to

the lower worm densities.

Even though the earthworms fed on ipomoea

leaves in reactors which also had soil and sand, and

produced a considerable amount of vermicast in all

the runs, there was a steady decrease in zoomass in

all ipomoea-based reactors. It is likely that the

ipomoea employed in these reactors had

accumulated toxic inorganics or heavy metals as it

was picked up from near high ways and chemical

industries. This might have caused the adverse

impacts. But as expected, cow manure was

preferred by earthworm over neem and ipomoea.

3.4. Surface area of vermicast

The surface area of the castings varied

significantly between feed to feed and was also

influenced by the presence of sand/soil (p<0.001)

(Table 2). The castings produced from reactors fed

with ipomoea, sand and soil had a significantly

higher surface area in comparison to others of that

set, and ranged from 16730.3±2384.2 to

17202.735±2069 µm2. Cow manure and neem fed

earthworms produced castings with surface areas

59

Figure 2. Percentage of sand and soil particle entrapped in the castings measured by gravimetric method

(mean ± SD) and percentage of sand particle covered area in the castings measured by thin-sectioning

method (mean ± SD). (‘w’ represent the worm density per kg substrate).

ranging 10297.8±171.8 to 13016±649.57 µm2 and

10112.5±2519.7 to 14125.5±1053 µm2,

respectively. The casts produced from the reactors

without sand and soil was significantly smaller in

size: 8651.2±1299.3µm2 and 7991.6±1537.6 µm

2

for cow dung and neem respectively. The size of

the castings positively correlated with the amount

of sand/soil ingested by the earthworms. The

difference in worm density did not show any

variable effect on cast size.

3.5. Assimilation of sand and soil particles in

castings

The percentage of sand and soil assimilation

in castings and the percentage of sand covered area

in vermicast sections are presented in Figure 2. A

clear difference can be seen between the amounts

of sand assimilated in castings of different feeds (p

< 0.001) (Table 2). Castings from ipomoea had the

largest sand covered area (Figure 3) of 17.7±1.17%

to 26.2±3.71%, followed by cow dung and neem

ranged from 8.93±3.33 to 13.8±2.06% and

6.57±0.27% to 10.1±2.29%, respectively. The

extent of sand assimilation in castings of cow-

manure and neem-fed earthworms differed slightly

but the difference was not statistically significant.

A similar trend was revealed by gravimetric

analysis of sand and soil entrapped in the vermicast.

The castings from ipomoea-fed reactors had

51.09±3.2%, 43.26±3.3% and 47.67±3.3% of sand

and soil when earthworm densities were 25, 15 and

5, respectively. The corresponding figures for cow

dung and neem vermicast were 26.79±3.6%,

24.23±0.6%, 22.73±1.6%, and 30.93±3.6%,

28.67±1.4%, 29.05±3.3%, respectively. As for

control reactors which has only soil and sand the

castings could not be harvested intact from the

reactors and hence their shape and size could not be

quantified. Irrespective of the size of the castings,

the area covered in it by sand differed significantly

in different treatments. The highest sand-covered

area was observed in castings from ipomoea:

9473.18 µm2. With cow manure and neem feed it

was 6047.72 and 5392.54 µm2 respectively; the

difference not being statistically significant. In the

present studies the earthworms were given access to

0

10

20

30

40

50

60

5w 15w 25w 5w 15w 25w 5w 15w 25w

Neem Cowdung Ipomoea

San

d/s

oil

en

trap

pe

d in

cas

tin

gs (

%)

Treatments

Sand and soil entrapped (%) Sand grains covered area (%)

60

Figure 3. The thin sectioned vermicast from neem, cow dung and ipomoea based reactors with soil + sand

bedding illustrated in a, b and c. The light – colored portions inside the castings are mineral soil particles.

The castings from the reactor without soil/sand showing in d.

Figure 4. Regression analysis between the sand and soil entrapped in the castings (%) and (a) the average

zoomass changes and (b) mortality rate.

y = -100.4x + 37.204 R² = 0.9201

0

10

20

30

40

50

60

-0.15 -0.1 -0.05 0 0.05 0.1 0.15 0.2

San

d a

nd

so

il e

ntr

app

ed

in c

asti

ngs

(%

)

Average zoomass changes worm-1 run-1

y = 1.3506x + 23.029 R² = 0.8916

0

10

20

30

40

50

60

0 5 10 15 20

San

d a

nd

so

il e

ntr

app

ed

in c

asti

ngs

(%

)

Average mortality (%)

61

both sand and soil and separate studies on sand or

soil were not carried out. But it can be surmised

that broad trends vis-a-vis pattern of ingestion

would be no different than seen in the present

study. When anecics ingest sand along with plant

debris and grind their feed, the attrition caused by

sand results in very fine fragmentation of organic

matter, enhancing its surface area and making it

much more amenable to microbial action than was

otherwise possible (Schulmann and Tiunov, 1999).

Had a similar process been occurring in the

presence study, the ingestion of sand and soil along

with neem and cow dung would have facilitated

digestion of the phytomass. But such was not the

case and there was no significant difference in

zoomass gain between reactors with and without

soil/sand bedding. The increase in sand

consumption rate in ipomoea-fed reactors may be to

fulfill their energetic requirements and avoid

consumption of ipomoea leaves. This tendency has

been seen in several other species (Curry and

Schmidt, 2007). A regression analysis between the

amount of sand and soil consumed and average

zoomass change in earthworms and mortality rate

substantiates these findings (Figure 4).

4. Conclusions

Controlled experiments were carried out to

see whether the epigeic earthworm – E. eugeniae

would ingest sand and soil even when phytomass

was available in abundance. It was seen that even

though initially E. eugeniae did ingest sand and soil

despite the luxury availability of phytomass, this

tendency was reduced as the time passed indicating

adaptive response to the phytomass feed.

These findings are of significance in

vermireactor’s design and optimization because

they indicate that sand-soil-gravel bedding as used

in conventional vermireactors is not really

necessary to ensure the survival, growth and

fecundity of the epigeics – used in the

vermicomposting. The findings reinforced the

validity of the high-rate vermicomposting concept

and associated nuances of vermireactors design

introduced earlier by the authors.

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Saxena, M., Chauhan, A., Asokan, P., 1998. Fly ash

vemicompost from non-friendly organic

wastes. Pollun. Res. 17, 5–11.




Selvamuthukumaran, D., Neelanarayanan, P., 2012.

Bio-conversion of leaves litter into

vermicompost by indigenous earthworm,

Perionyx excavates. Electron. J. Environ. Sci.

5, 55-60.




fertilizer production. Environ. Sci. Pollut. Res.

21, 1349-1359.

Sim, E.Y.S., Wu, T.Y., 2010. The potential reuse of

biodegradable municipal solid wastes (MSW)

as feedstocks in vermicomposting. J. Sci. Food

Agric. 90, 2153-2162.

Suthar, S., Singh, S., 2008. Feasibility of

vermicomposting in biostabilization of sludge

from a distillery industry. Sci. Total Environ.

394, 237-243.

Suthar, S., 2014. Toxicity of methyl parathion on

growth and reproduction of three ecologically

different tropical earthworms. Int. J. Environ.

Sci. Technol. 11, 191-198.

Tauseef, S.M., Abbasi, T., Banupriya, D.,

Vaishnavi, V., Abbasi, S.A., 2013.

HEVSPAR: A novel vermireactor system for

treating paper waste, Off. J. Pat. Off. 24,

12726.

Tian, G., Olimah, J.A., Adeoye, G.O., Kang, B.T.,

2000. Regeneration of earthworm populations

in a degraded soil by natural and planted

fallows under humid tropical conditions. Soil

Sci. Soc. Am. J. 64, 222-228.

Wang, L., Zhang, Y., Lian, J., Chao, J., Gao, Y.,

Yang, F., Zhang, L., 2013. Impact of fly ash

and phosphatic rock on metal stabilization and

bioavailability during sewage sludge

vermicomposting. Bioresour. Technol. 136,

281-287.

FEEDING BEHAVIOUR OF PHYTOPHAGOUS

EARTHWORM EUDRILUS EUGENIAE IN HIGH-

SUBSTRATE COLUMN VERMIREACTORS

Chapter

4

65

A paper based on this chapter has been

submitted for publication.

CChhaapptteerr 44

Feeding behaviour of phytophagous earthworm Eudrilus

eugeniae in high-substrate column vermireactors

Abstract

In chapter 3 study made to check whether the epigeic earthworm, Eudrilus eugeniae will ingest soil/sand

even when there is availability of phytomass in plenty is reported. It was seen that the worms, ingested not

only soil even sand particles along feed. Moreover, the soil/sand ingestion rate was significantly influenced

by the substrate type – cow manure, leaves of neem and of ipomoea – tested by us. Since, the findings have

important implications in the design of vermireactors for maximizing their efficiency; the influence of

vemireactors’ constituent on the epigeic’s feeding behavior is of the essence, which needs to be resolved. In

this chapter, study on the influence of (a) feed height in the vermireactors (b) the mode of supply of feed, and

(c) the degree of decomposition of the feed, on the preference towards sand and soil by the epigeic

earthworm, E. eugeniae is reported.

1. Introduction

Earthworms are one of the most important

component of soil biota in terms of soil formation

and maintenance of soil structure and fertility

(Edwards, 2004). The function of earthworms on

soil ecosystem differs considerably with different

earthworm species that are linked to their feeding

strategy (Uchida et al., 2004). The earthworms

mainly feed on organic matter together with

microorganisms, micro and meso fauna, nematodes

and their dead remains (Curry and Schmidt, 2007).

However, anecics and endogeics species which

often live in subsoil, consume an appreciable

amount of mineral soil, even in the presence of

phytomass and animal manure (Hendriksen, 1991;

Schulmann and Tiunov,1999), possibly to keep

their gizzard muscles toned up (Marhan and Scheu,

2005). The epigeics are considered to ingest only

organic matter in various degrees of decay.

As described in chapter 3, study was

conducted to check whether the epigeic

earthworm, Eudrilus eugeniae will ingest soil/sand

even when there is luxury availability of

phytomass. It was seen that initially the E. eugeniae

did ingest sand and soil like anecic and endogeic

species, despite the availability of phytomass in

plenty. The amount of soil/sand ingestion

significantly varied with different feed – leaves of

neem and of ipomoea, and cow manure. As the

days progressed, the consumption of sand and soil

decreased with all the three feed. Influence of

different earthworm densities on soil/sand

consumption was also studied, and it did not show

any variable effect on sand /soil consumption.

Since these studies have important

implications in the design of vermireactors for

maximizing their efficiency, it is necessary to

understand the influence of vemireactors’

constituent on the epigeic’s feeding behavior.

Hence in the present study, the feeding activity of

the epigeic earthworm, E. eugeniae was tested in

high-substrate column vermireactors by varying the

parameters such as: (i) substrate height, (ii) degree

of decomposition of the substrate, and (iii) mode of

substrate input. Neem leaves, which are locally

66

available in the experimental area, were used as

feeding substrate for the earthworms.


Neem leaves collected from the Pondicherry

University campus and its vicinity were washed

with water to remove adhering particles. The

vermireactors of 35 cm width, 39 cm length and

100 cm height were prepared with basal layer of

coarse sand to a thickness of 3 cm followed by 5

cm thick layer of soil. Since the substrate height

was up to 60 cm in the experimental reactors, all

the four sides of the reactor were fabricated with

nylon mesh of pore size of 0.3 mm supported by a

wooden frame to avoid anaerobic condition set in.

Two sets of duplicate reactors were prepared in this

manner. In the first set, the reactors were filled with

neem leaves up to 45 and 60 cm height over sand

and soil bedding respectively (T1 and T2

treatments). In each reactor, 10 numbers of adult

worms of similar size were introduced after

determination of their biomass. The duration of

each run was 15 days. At the end of each run,

vermicast generated in the reactors were harvested

and the reactors were restarted with the leftover

substrate with adult worms, with which the reactors

were started. To maintain the initial substrate

height, fresh leaves were added to the left over

substrate whenever the height reduced. To maintain

the initial number of worms, hatchlings produced, if

any, were removed at the end of each run.

In order to study the influence of degree of

decomposition of substrate on epigeic preference

towards sand and soil, the second set of reactors

were prepared with 45 and 60 cm of substrate

height respectively (T3 and T4 treatments). At the

end of each run, the vermicast was harvested and

restarted with left over substrate from the previous

run. In these reactors, to study the influence of

increase in worm density on epigeic feeding

activity, juveniles and cocoons generated in the

previous run were reintroduced along with adult

worms after assessment. One set of reactors was

maintained as control, which was prepared with 45

cm height substrate and without soil and sand

bedding. The control reactor was also operated

similar to the second experimental set.


ambient conditions with temperature of 29 ± 4°C.

Moisture content of substrate was maintained about

60% throughout the study period by periodic

sprinkling of adequate quantity of water. At the end

of each run, the castings were carefully separated

from other particles by soft painting brush and

quantified. While disbanding the reactors, the adult

earthworms with which the experiment was started,

were washed, blotted dry and weighed without

emptying their gut to record their zoomass and

introduced back into the respective reactors. The

dead earthworms, if present, were replaced, while

starting the next run.


in the vermicast was quantified by gravimetric

method. Samples oven dried at 105°C to a constant

weight were put in distilled water, and crushed to

release the soil/sand. The content was filtered

through a Whatman No. 42 filter paper. The residue

over the filter paper was kept in a muffle furnace at

550°C for 4 hours to remove organic matter present

in the castings (John, 2004). Petrographic thin

sections were made of air-dried vermicast using

Bueller PetroThin® thin sectioning system

(Buehler, Lake Bluff, IL, USA) after impregnating

them in araldite and xylene (solvent) mixture. The

resulting specimens were observed under polarizing

microscope to confirm the sand assimilation in the

vermicast (FitzPatrick, 1993).

The data was statistically analyzed by two-

way ANOVA to assess the impact of

presence/absence of sand and soil vermibed on

vermicast production, earthworm mortality,

zoomass gain and assimilation of sand and soil

content in the vermicast. The post hoc test LSD has

been done to find the significant difference between

each subject. All statistical analyses were

67

performed using the statistical software SPSS 16

package.


The reactors T1, T2 and control were

operated for 12 runs by continuous feeding of

substrate at each run. In the T3 and T4 treatments,

substrate height of the reactors reduced to 5 cm

during the ninth run, hence these reactors were not

operated further. There was a significant difference

in the amount of sand and soil assimilated in the

vermicast with different treatments (p<0.001). The

soil assimilation rate was high in the initial runs

with all the treatments (Figure 1). In the first three

runs, the percentage of sand and soil content in the

reactors of T1 and T2 was 36.5 and 29.7% on

average, and those of T3 and T4 treatments were

28.4 and 24.0%, respectively. The high soil

assimilation rate in the initial runs was probably

due to presence of polyphenolic compounds in the

fresh neem leaves. A negative correlation between

the palatability of leaf litter and its total polyphenol

content of substrate demonstrated in previous

studies (Satchell and Low, 1967; Hendrickson,

1990; Tian et al., 2000), may be the reason for

earthworm preference towards sand and soil during

the initial runs. From the fourth run there was a

steady decline in assimilated sand and soil content

in the castings of both T1 and T2 treatments, which

might have attributed to the degradation of phenolic

components in the substrate. The previous study by

Gajalakshmi and Abbasi (2004) also support this

assumption, in which earthworms voraciously fed

the neem leaves after pre-composting.

In the T1 and T2 treatments, the sand and soil

assimilation rate was stabilized from the ninth run,

in which less than 5% of sand and soil content was

recorded with most of the cases. In the case of T3

and T4 treatments, the sand and soil assimilation

showed a declining trend up to fifth run and

subsequent runs showed increase in soil and sand

assimilation in the vermicast. Due to continuous

harvesting of vermicast at the end of each run, the

substrate height of these reactors steadily declined

throughout the experimental period, which might

have attributed to the high sand and soil

assimilation after fifth run. At ninth run, 33.3 and

17.4% of soil and sand assimilation was recorded in

the T3 and T4 treatments, and the substrate height

of these reactors were 6 and 9 cm, respectively at

the start of the ninth run. However, during the first

five runs, the sand and soil assimilation rate in the

Figure 1. Percentage of sand and soil particles entrapped in the castings of different treatments

0

10

20

30

40

3 4 5 6 7 8 9 10 11 12

Soil

and

san

d e

ntr

app

ed in

cas

tin

gs

(%)

Runs

T1

T2

T3

T4

68

Figure 2. Vermicast output, grams worm -1

day-1

recorded in reactors with different treatment.

T3 and T4 treatments were much lower than the T1

and T2 treatments. This indicates that the

earthworms did not prefer sand and soil, when the

degraded substrate is available to them. The initial

height of the substrate in the reactors did not show

any noticeable difference in sand and soil

assimilation in vermicast. The assimilation of sand

and soil in the castings was confirmed with

petrographic thin sections.

The different heights of the substrate did

not have any significant impact on vermicast

output, but the mode of substrate input (either one

time or continuous supply of substrate) has changed

the vermicast output considerably (Figure 2). But

the statistical analysis did not show any significant

difference within different treatments due to much

variation in data. During the first three runs, very

small quantity of vermicast was produced in all the

reactors. At the end of third week, the vermicast

output was 0.525 and 0.280 g worm-1

d-1

with T1

and T2 treatments, and those of T3 and T4 showed

0.805 and 0.215 g worm-1

d-1

, respectively. In

subsequent runs, there was increase in vermicast

output with all the treatments. The increase in

vermicast output was much higher in the T3 and T4

treatments during the first six runs; in some cases it

was about twofold higher than the T1 and T2

treatments. The reason for increase in vermicast

output in T3 and T4 treatments may be probably

due to increase in the degree of decomposition of

the substrate they contained. But, in the T1 and T2

treatments, the addition of fresh substrate at each

run to maintain the substrate height might have

increased the polyphenolic content of the whole

substrate which in turn reduced the vermicast

output. Studies on acacia (Acacia auriculiformis)

(Ganesh et al., 2009), oak, beech, larch, spruce

(Satchell and Low, 1967) and common St. John's

wort (Hypericum perforatum) (Schonholzer et al.,

1998) also shows that the freshly fallen leaves of

these plants are unacceptable by the worms due to

their phenolic and other toxic compounds. Further

runs, did not show any difference in vermicast

output with these treatments probably due to

acclimatization of the earthworms to this substrate.

Throughout the study, the vermicast output

from the control treatment was very close to the T1

treatment which comprised similar substrate height.

The changes in earthworm zoomass and mortality

also did not show any significant difference

between different treatments. During the first three

runs, there was loss in earthworm biomass and high

mortality was observed with all the treatments;

thereafter there was no significant difference in any

0

0.5

1

1.5

2

2.5

3 4 5 6 7 8 9 10 11 12

Ver

mic

ast

ou

tpu

t, g

wo

rm -1

d-1

Runs

T1

T2

T3

T4

Control

69

of these parameters. The overall results show that

existence of sand and soil bedding neither

influenced the vermicast output nor their growth or

mortality with neem as a feed.

4. Conclusions

The findings of the present study clearly

shows that the diet of the epigeic worm E. eugeniae

consist of considerable portion of soil and sand in

the initial period. However, the amount of soil and

sand ingestion rate drastically reduced as the days

progressed. The earthworm adaptability to neem

leaves might have reduced the sand and soil

ingestion rate. Varying substrate height of the

reactors did not show any influence on mineral soil

ingestion, provided the height was not less than 5

cm. However, ingestion of soil and sand particles

neither enhanced the E. eugeniae’s digestion

process nor feeding rate or its growth as reported

with anecic and endogeic earthworms.

References

Bolton, P.J., Phillipson, J., 1976. Burrowing,

feeding, egestion and energy budgets of

Allolobophora rosea (Savigny) (Lumbricidae).

Oecologia 23, 225-245.

Bostrom, U., 1988. Ecology of earthworms in

arable land. Population dynamics and activity

in four cropping systems. PhD thesis. Report

No. 23. Swedish University of Agricultural

Science, Uppsala.

Boyle, J., 2004. A comparison of two methods for


sediments. J. Paleolimnology 31, 125-127.


of earthworms - A review. Pedobiologia 50,

463-477.


CRC Press, Washington, D.C.



York.



vermicompost. Bioresou. Technol. 92, 291-296.

Ganesh, P.S., Gajalakshmi, S., Abbasi, S.A., 2009.

Vermicomposting of the leaf litter of acacia

(Acacia auriculiformis): Possible roles of

reactor geometry, polyphenols, and lignin.

Bioresou. Technol. 100, 1819-1827.



Fert. Soils 10, 17-21.

Hendriksen, N.B., 1991. Gut load and food-

retention time in the earthworms Lumbricus

festivus and L. castaneus: a field study. Biol.

Fert. Soils 11, 170-173.



decomposition in soil and in casts of Lumbricus

terrestris L. Geoderma 128, 155-166.

Martin, N.A., 1982. The interaction between

organic matter in soil and the burrowing

activity of three species of earthworms

(Oligochaeta:Lumbricidae). Pedobiologia 24,

185-190.

Satchell, J.E., Low, D.G., 1967. Selection of leaf

litter by Lumbricus terrestris, in: Graff, O.,

Satchell, J.E. (Eds.), Progress in Soil Biology.

North Holland Publi. Co, Amsterdam, pp. 102-

119.

Schonholzer, F., Kohli, L., Hahn, D., Daniel, O.,

Goez, C., Zeyer, J., 1998. Effects of

decomposition of leaves on bacterial biomass

and on palatability to Lumbricus terrestris L.





Tian, G., Olimah, J.A., Adeoye, G.O., Kang, B.T.,

2000. Regeneration of earthworm populations

in a degraded soil by natural and planted

fallows under humid tropical conditions. Soil.

Sci. Soc. Am. J. 64, 222-228.

Uchida, T., Kaneko, N., Ito, M.T., Futagami, K.,

Sasaki, T., Sugimoto, A., 2004. Analysis of the

feeding ecology of earthworms

70

(Megascolecidae) in Japanese forests using gut

content fractionation and δ15N and δ13C stable

isotope natural abundances. Appl. Soil Ecol.

27, 153-163.

EFFECT OF SAND AND S

PHYTOPHAGOUS EARTHWO

EUGENIAE ON THE PHYSICAL AND

PROPERTIES OF VERMIC

EFFECT OF SAND AND SOIL INGESTION BY

PHYTOPHAGOUS EARTHWORM EUDRILUS

ON THE PHYSICAL AND CHEMICAL

PROPERTIES OF VERMICAST

OIL INGESTION BY

EUDRILUS

CHEMICAL

Chapter

5

71



CChhaapptteerr 55

Effect of sand and soil ingestion by phytophagous earthworm

Eudrilus eugeniae on the physical and chemical properties of

vermicast

Abstract

The studies reported in chapters 3 and 4 have revealed that even if sand/soil are not required for the epigeics

growth and survival, the earthworms prefer sand/soil with fresh phytomass. This tendency subsided once the

earthworms got acclimatized to the feed. However, the effect of sand and soil ingestion on the properties of

vermicast is not yet explored. Hence, a controlled experiment was carried out with Eudrilus eugeniae in

vermireactors with different masses of substrate, to study the feeding behavior of epigeics and its impact on

the properties of vermicast they generate. The results reinforce our earlier finding that the epigeics ingest soil

and sand inspite of the luxury availability of phytomass, but the consumption of sand and soil neither

facilitate their survival, growth and fecundity of the epigeics. Ingestion of sand and soil had increased the

bulk density and particle density of the vermicast in addition to reduction of pore space, water retention

capacity, and nutrient content.

1. Introduction

Of all the larger inhabitants of the soil,

probably none is more important than the

earthworm in terms of soil formation and

maintenance (Carson, 1962). The earthworms

significantly affect the soil structural characteristics

and distribution of resources to many soil animals

and plants mainly through the castings they

produce. The castings exists in various forms and

features which is determined by earthworm species,

their habitat and feeding behavior. Generally the

castings are enriched in organic matter, nutrients

and foster high levels of microbial activity due to

the selective foraging of organic particles (Fonte et

al., 2007). In addition, anecic and endogeic

earthworm groups defined by Bouche (1977),

ingest considerable portion of soil/sand along with

organic matter and produce complex organo-

mineral structures as castings. Although, ingestion

of soil/sand particulate by earthworms facilitate

mechanical fragmentation of organic matter during

gut transition (Schulmann and Tiunov, 1999),

assimilation of soil particles in castings alter

surface soil texture and other physical properties

(Lavelle and Spain, 2001; Blanchart et al., 2004;

Jouquet et al., 2008a,b), microbial activity and

diversity (Marhan, 2004), and therefore the

mineralization or sequestration of soil organic

matter and the retention or the leaching of mineral

nutrients (Brady and Weil, 1999; Lal, 2004).

Although a number of studies have discussed

these aspects, the information are limited to anecic

and endogeic groups of earthworms. Even though,

the epigeics are reported to ingest soil and sand in

few studies (Martin, 1982; Bostrom, 1988;

Karthikeyan et al., 2014), less is known about the

impact of soil ingestion on their digestion, nutrient

uptake and properties of vermicast. Studying this

aspect is particularly important to understand the

role of epigeics in soil processes as well as to

enhance the nutrient cycle of organic matter in both

agricultural and waste management systems.

72

Therefore, in the present study, an attempt has been

taken to understand the impact of feeding activity

of the epigeic earthworm, Eudrilus eugeniae on

their growth and properties of vermicast produced

by them. The earthworms were grown with neem

leaves as feed, in the presence/absence of soil and

sand with different quantities of the substrate. The

presence of sand and soil in the vermicast was

confirmed by petrographic thin section method. The

amount of soil and sand assimilated in the castings

was quantified by both gravimetric and wavelength

dispersive X-ray fluorescence spectrometry

methods. The effect of soil/sand ingestion on the

growth of earthworm and vermicast production had

been monitored in the neem fed vermireactors for

240 days. The difference in physical and chemical

properties of vermicast generated from reactors

supplied with or without soil/sand was also studied

and the results are discussed in brief.


2.1. Experimental design

Neem leaves were collected from the

Pondicherry University campus and its vicinity.

The leaves were washed with water to remove

adhering particles. Rectangular, 41 liter wooden

boxes (30 cm high with surface area of 35 x 39 cm)

were used as vermireactors. They were lined up

with thick transparent plastic sheets to make

reactors impermeable, prevent earthworms escaping

from the reactors and also to protect them from

predators. Four sets of duplicate vermireactors were

employed. First and second set of duplicate reactors

were filled with 250 and 500 g dry weight

equivalent of neem leaves (T1 and T2 treatments).

The third and fourth sets of reactors were prepared

with basal layer of coarse sand to a thickness of 3

cm followed by a 5 cm thick layer of soil. Over it

250 and 500 g dry weight equivalent of neem

leaves was placed (T3 and T4 treatments). Ten

adult earthworms having approximately equal size,

randomly picked from their cow dung-based

culture, were introduced in each of the reactors.

2.2. Vermireactors operation


ambient conditions with a temperature range of

29±4°C. Moisture content of substrate was

maintained about 60% throughout the study period

by periodic sprinkling of adequate quantity of

water. The duration of each run was 15 days. At the

end of each run, vermicast mass, mortality and

changes in the earthworm zoomass were

determined, and then the reactors were restarted

with the leftover substrate with adult worms, with

which reactors were started. To maintain the initial

number of worms, hatchlings produced, if any,

were removed and dead worms were replaced,

while starting the next run. During the first trial, the

reactors were operated for 135 days. The second

trial comprised of 105 days, which had been

operated with newly assembled reactor contents

and earthworms from the previous trial.

2.3. Analytical methods

Moisture content of castings was determined

by weight loss at 105°C. The bulk density and

particle density of samples was determined

according to Bashour and Sayegh (2007). To

measure the water holding capacity, the samples

were filled in cylinders with a perforated base and

immersed in water and drained. The quantity of

water taken up by samples is determined by drying

to constant mass at 105°C (Margesin and Schinner,

2005). The total porosity and water filled pore

space (WFPS) were calculated from the particle and

bulk density values of the respective samples

(Carter and Gregorich, 2008). The electrical

conductivity (EC) was measured with sample

suspension of 1:2 (w/v) by using EI™ 611E EC

meter (Bashour and Sayegh, 2007).

Total organic (Corg) content in the castings

was measured by modified dichromate redox

method (Heanes, 1984). In this method, external

heating was applied during the oxidation process in

order to quicken and complete oxidation of Corg in

73

the sample. Total nitrogen (Ntot) content was

determined by modified Kjeldahl method

(Kandeler, 1993) using Kel Plus™ semi-automated

digester and distillation units (Elico Ltd,

Hyderabad, India). In order to include nitrate,

nitrite, nitro and nitroso groups, a mixture of

salicylic acid and sulfuric acid was used for

digestion. The SiO2, potassium and phosphorus

content in the vermicast were determined by

Bruker™ S4-Pioneer model wavelength dispersive

X-ray fluorescence spectrophotometer (WD-XRF,

Bruker, Billerica, MA, USA). The samples were

ground to particle size well below 100 µm using

ball mill in order to minimize the grain size

interference on XRF-measurement.

2.4. Assessment of soil/sand content in the

vermicast


in the vermicast was quantified by gravimetric

method. Samples oven dried at 105 °C to a constant

weight were put in distilled water, and crushed to

release the soil/sand. The content was filtered

through a Whatman No. 42 filter paper. The residue

over the filter paper was kept in a muffle furnace at

550°C for 4 hours to remove organic matter present

in the castings (John, 2004). Petrographic thin

sections were made of air-dried vermicast using

Bueller PetroThin® thin sectioning system

(Buehler, Lake Bluff, IL, USA) after impregnating

them in araldite and xylene mixture. The resulting

specimens were observed under a polarizing

microscope to confirm the sand assimilation in the

vermicast (FitzPatrick, 1993).

2.5. Data analysis

The influence of sand and soil ingestion on

the growth and survival of earthworm, vermicast

mass and its physico-chemical properties were

assessed by independent sample t-test. The

statistical calculation was carried out using SPSS

16 software.


3.1. Assimilation of sand/soil in the vermicast

The percentage of sand and soil entrapped in

the earthworm castings in each run of 15 days

duration with different treatments is shown in

Figures 1 and 2. There was a significant difference

in the amount of sand/soil assimilation in the

vermicast from the reactors with different amount

of substrate and duration of reactor operation (p

<0.001).

Figure 1. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil with 250

g (T3a and T3b, duplicates) and 500 g substrate (T4a and T4b, duplicates) in the first trial.

0

15

30

45

60

75

3 4 5 6 7 8 9

Soil

and

san

d e

ntr

app

ed in

VC

(%

)

No. of runs

T3 a T3 b T4 a T4 b Linear (T3 b)

74

Figure 2. Percentage of sand and soil entrapped in the vermicast from reactors consisting sand/soil with 250

g (T3a and T3b, duplicates) and 500 g substrate (T4a and T4b, duplicates) in the second trial.

In both the first and second trials, high sand/soil

assimilation was recorded during the initial run

with all the treatments. During the first three runs,

the percentage of sand and soil content in the

reactors of T1 was 60.7% on average, and those of

T2 treatments were 47.0%, respectively. There was

a steady decline in sand/soil assimilation with all

the treatments up to 8th and 6

th run of first and

second trials, respectively. The high sand/soil

assimilation in the initial run was probably due to

the presence of some of the unaccepted substance

in fresh neem leaves like phenolic and other toxic

compounds. Previous reports on leaves of many

tree species, includes oak, beech, larch, and spruce,

which are reported to contain phenolic compounds,

were also shown unpalatable by earthworms

(Satchell, 1967). Similarly, studies reported by

Hendriksen (1990) and Schonholzer et al. (1998)

revealed that the palatability of different kinds of

leaf litter by Lumbricus spp. and Aporrectodea spp.

is largely determined by the C:N ratio, lignin and

the polyphenol concentration of leaf litter.

Reduction in the assimilation of sand/soil in the

vermicast in further runs indicates that degradation

of the unacceptable substance in fresh leaves has

been initiated. The percentage of sand/soil

assimilation was reduced to about 20% in the 8th

and 6th run with first and second trials, respectively.

The percentage of SiO2 in the vermicast

significant by varied with different reactors

consisting different amounts of substrate (Table 1).

The percentage of SiO2 was 40.4% in the castings

from T1 treatment, and those of T2 were 24.4%,

respectively. In the first trial, the gravimetrically

determined sand/soil content in the castings was

higher in reactors of T1 treatment than those of T2

treatment during the first eight runs. From ninth run

onwards there significant difference in amount of

sand/soil assimilated in the castings from different

treatments. During the second trial, the assimilation

of sand/soil in the castings had stabilized on the

sixth run probably due to acclimatization of the

earthworms to this substrate. However, all the

reactors were showing slight increase in soil/sand

assimilation in last few runs. Due to continuous

harvesting of vermicast at the end of each run, the

substrate height of these reactors steadily declined

drastically, which might have attributed to the high

sand/soil assimilation in last few runs.

0

15

30

45

60

75

2 3 4 5 6 7

Soil

and

san

d e

ntr

app

ed in

VC

(%

)

No. of Runs

T3 a T3 b T4 a T4 b Linear (T3 b)

75

3.2. Vermicast output

Presence of sand/soil bedding showed

significant influence on vermicast production, but

the difference in the substrate quantities did not

show any noticeable changes in vermicast output

(Figure 3). During the initial runs, there was very

less quantity of vermicast production with all the

reactors. At the end of third week, the vermicast

output was 0.318±0.046 g worm-1

day-1

with

reactors without bedding, and that of reactors with

bedding showed 0.482±0.048 g worm-1

day-1

.

Fourth and fifth runs showed an increase in

vermicast output with all the treatments, and it was

about fivefold in some of the reactors. The drastic

increase in vermicast output may be due to increase

in the degree of decomposition of the substrate they

contained. As discussed in the previous section, the

degradation of polyphenolic and other toxic

compounds in the neem leaves would have

increased the vermicast output in all the treatments.

The vermicast output in subsequent runs, did

not show any significant difference while the height

of the substrate was not less than 1 cm. But in all

the runs, the production of vermicast was higher in

the reactors with bedding than without bedding

which indicates that the presence of bedding had

significant influence on vermicast output. However,

the vermicast from the reactors with bedding

consisted of a considerable portion of sand/soil.

There was no significant difference observed with

both trails, when the vermicast output of reactors

without bedding was compared with reactors

consisting bedding after subtracting the amount of

assimilated sand/soil with vermicast output. This

indicates that the ingested amount of substrate

alone by earthworms was not influenced by the

presence of bedding.

3.3. Growth and survival of earthworms

The presence of bedding and amount of

substrate input in vermireactors did not show any

significant influence on the mortality and changes

in earthworm zoomass. The first two runs of first

trial showed reduction in zoomass in most of the

reactors; whereas in the second trial, the zoomass

reduction was observed only in the first run. In the

first trial, the reduction in zoomass was

0.036±0.010 and 0.026±0.012 g worm-1

, and in the

Figure 3. Vermicast output, grams worm -1

day-1

(mean ± SD) recorded in reactors without sand/soil + 250

and 500 g substrate (T1 and T2 treatments) and reactors consisting sand/soil + 250 and 500 g substrate (T3

and T4 treatments).

0

1

2

3

a b a b a b a b

T1 T2 T3 T4

Ver

mic

ast

ou

tpu

r g

wo

rm-1

d-1

Treatments

1st Trial

2nd Trial

Linear (2nd Trial)

76

second trial, 0.039±0.022 and 0.050 ±0.017 g

worm-1

at the end of the first run in the reactors with

the presence and absence of bedding, respectively.

The subsequent runs of both trials showed increase

in zoomass in most of the cases. Likewise, high

earthworm mortality was also recorded in the initial

runs with both the trials. In the first trial, the

reactors without bedding showed a maximum of

50% mortality with T3 treatment. The reactors with

bedding showed a maximum mortality of 40% in

the T2 treatment. Similarly, in the second trial, a

maximum of 30% mortality was observed in one of

the reactors without bedding, and that of a reactor

with bedding showed 20% mortality, respectively.

Except the last run of both the trials, there was no

significant mortality. The last run of both trials

showed reduction in zoomass and mortality in some

of the reactors, probably due to decline in substrate

availability in these reactors. Similarly, increase in

the mortality of earthworms with few or no organic

residues added soil has been already reported in

both pot and field trials (Edwards, 2004).

3.4. Physical and chemical properties of

vermicast

The physical properties of vermicast significantly

varied (p <0.001) with presence or absence of

bedding. In the reactors without bedding, the

amount of substrate had no impact on the properties

of vermicast, but it exhibited significant influence

when the reactors consisted bedding. The presence

of bedding increased the bulk and particle density,

and reduced the total porosity, WFPS and WHC of

the vermicast (Table 1). The average bulk density

of castings from the reactors of both T3 and T4

treatments was 0.303 g cm-3

, whereas it was 0.672

and 0.467 g cm-3

in the T1 and T2 treatments,

respectively. In the case of particle density, the T3

and T4 treatments showed 1.145 g cm-3

, and those

that of T1 and T2 was 1.706 and 1.441 g cm-3

,

respectively. Assimilation of high density soil

particle in the vermicast might be the reason for

higher bulk and particle density of castings from

reactors comprised of bedding. Studies on anecic

earthworms, Martiodrilus carimaguensis and

Lumbricus terrestris with different soil types have

also shown increase in density of castings when the

mineral soil consumption was high (Decaëns, 2000;

Josehko et al., 1989). Fluctuation in the bulk and

particle density of castings from the reactors

consisting different quantities of substrate were

probably due to the variation in sand/soil

assimilated by the earthworms in these reactors.

The total porosity and WFPS of castings were also

Table 1. The physical properties of vermicast generated from the reactors without sand/soil + 250 and 500 g

substrate (T1 and T2 treatments) and reactors consisting sand/soil + 250 and 500 g substrate (T3 and T4

treatments). The alphabets ‘a’ and ‘b’ represent duplicate reactors.

Treatments Water Content

%

Particle Density

g cm-3

Bulk Density

g cm-3

Pore Space

% WFPS % WHC %

EC

mmhos cm-1

T1-a 72.9±1.8 1.139±0.004 0.288±0.006 74.7±0.5 24.7±1.2 525.8±10.9 2.20±0.05

T1-b 75.4±1.6 1.192±0.007 0.309±0.006 74.1±0.4 26.4±0.3 590.5±12.9 2.23±0.04

T2-a 68.0±2.5 1.127±0.008 0.303±0.006 73.2±0.6 25.0±1.2 614.4±11.4 2.16±0.03

T2-b 73.2±1.5 1.122±0.007 0.312±0.009 72.2±0.7 28.2±1.5 531.8±21.5 2.32±0.10

T3-a 33.2±0.4 1.709±0.018 0.709±0.013 58.5±0.3 40.1±1.2 204.3±27.3 1.66±0.16

T3-b 35.4±0.5 1.703±0.017 0.634±0.008 62.8±0.4 35.6±0.8 232.5±7.3 1.76±0.03

T4-a 42.0±0.9 1.511±0.013 0.496±0.014 67.2±1.0 30.8±1.2 350.7±6.7 1.69±0.03

T4-b 47.3±0.3 1.371±0.021 0.438±0.017 68.0±0.7 30.3±1.7 389.6±8.0 1.82±0.04

77

Table 2. The chemical properties of vermicast generated from the reactors without sand/soil + 250 and 500 g

substrate (T1 and T2 treatments) and reactors consisting sand/soil + 250 and 500 g substrate (T3 and T4

treatments). The alphabets ‘a’ and ‘b’ represent duplicate reactors.

Treatments TC mg g-1

TN % TK % TP % SiO3 %

T1-a 390.9±9.3 31.87±0.09 1.185±0.044 0.527±0.008 0.114±0.012

T1-b 388.0±4.3 34.06±0.11 1.180±0.046 0.473±0.011 0.092±0.005

T2-a 396.8±5.1 33.78±0.12 0.987±0.067 0.479±0.004 0.119±0.007

T2-b 389.9±3.5 33.32±0.04 1.032±0.046 0.521±0.005 0.099±0.008

T3-a 150.5±3.8 12.15±0.09 0.781±0.025 0.349±0.041 39.26±5.59

T3-b 139.8±7.7 15.33±0.20 0.705±0.017 0.428±0.026 41.47±6.66

T4-a 187.4±9.3 15.31±0.17 0.761±0.024 0.453±0.033 26.33±4.96

T4-b 186.8±7.5 14.87±0.15 0.842±0.017 0.365±0.029 22.54±2.70

significantly lower in the reactors with bedding

than the reactors devoid of bedding. The total

porosity of castings generated from the T1 and T2

treatments was 18.5 and 7.0% lower than the T3

and T4 treatments, respectively. Similarly castings

generated from the T1 and T2 treatments showed

about 60 and 34%, respectively lower WHC than

the castings from corresponding reactors without

soil/sand bedding. The lower WHC of the castings

in reactors comprised of bedding may be attributed

by the lower pore space of the castings (Chaudhuri

et al., 2009). In these treatments, the existence of

soil/sand particle in the castings reduced the

porosity and WHC of vermicast.

There was significant influence (p < 0.001)

on the chemical properties of vermicast with

reactors consisting of bedding. But the amount of

substrate had no impact on the properties of the

vermicast. The castings from the reactors without

bedding had maximum total C, N, P and K content

than the castings from reactors comprising of

bedding (Table 2). The castings from T3 and T4

reactors showed about 63 and 52% higher organic

carbon than the T1 and T2 reactors, respectively.

Similarly the nitrogen content was about 58 and

54% lower in the T1 and T2 reactors compared to

T3 and T4 reactors. The total potassium content of

castings was approximately 37% lower in T1

reactors than T3 reactors, and those of T2 reactors

showed 20% lower than T4 treatment, respectively.

A significant amount of sand and soil entrapped in

the castings may be the reason for the lower content

of C, N and K in castings from reactors with

sand/soil bedding compared to the ones without

bedding. Phosphorus content of castings has not

showed any significant difference between different

treatments. The phosphorus present in the soil/sand

entrapped in the castings might have attributed this

insignificant difference between these treatments.

4. Conclusions

The study gives inference that epigeic

earthworm – E.eugeniae also ingest sand and soil

like anecics and endogeics species. But once the

earthworms got acclimatized to the phytomass as

feed, the amount of soil and sand ingestion was

insignificant. Moreover, the ingestion of sand and

soil by epigeics, neither facilitated their digestion

nor its survival as there is no significant difference

in zoomass gain and earthworm mortality in

reactors with and without sand/soil bedding. There

was increase in the bulk density and particle

density, reduction in the pore space, water holding

capacity and nutrient content of vermicast in

reactors with soil/sand bedding. The overall finding

shows that the epigeic earthworm, E.eugeniae

78

really does not require sand/soil in their diet unlike

anecic and endogeic species. Moreover, the

physico-chemical characterization of vermicast

reveals that the assimilation of sand/soil in the

vermicast may reduce their beneficial impact on

plant growth and soil.

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in compacted soil with a combination of

morphological and soil physical measurements.

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Hanboonsong, Y., Brunet, D., Montoroi, J.P.,

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on soil properties of paddy fields in North-East

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climate change. Geoderma 123, 1–22.

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matter. Ph.D thesis. Technischen Universität,

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terrestris L. Pedobiologia 43, 453–458.

EFFECT OF VERMICAST GENERATED FROM AN

ALLELOPATHIC WEED LANTANA (LANTANA CAMARA)

ON SEED GERMINATION, PLANT GROWTH, AND

YIELD OF CLUSTER BEAN (CYAMOPSIS

TETRAGONOLOBA)

Chapter

6

81


Environmental Science and Pollution Research DOI: 10.1007/s11356-014-3103-5

CChhaapptteerr 66

Effect of vermicast generated from an allelopathic weed

lantana (Lantana camara) on seed germination, plant growth,

and yield of cluster bean (Cyamopsis tetragonoloba)

Abstract

In perhaps the first-ever study of its kind the effect of vermicompost, derived solely from an allelopathic

weed, on the germination, growth, and yield of a botanical species has been carried out. In test plots, the soil

was treated with the vermicompost of lantana (Lantana camara) at the rates of 5, 7.5 and 10 tons per hectare

(t ha-1

) and cluster bean (Cyamopsis tetragonoloba) was grown on it. The performance of these systems was

compared with the systems in which the soil was fortified with inorganic fertilizers (IF) in concentrations

equivalent to those present in the respective vermicompost (VC) treatments. Additionally a set of control was

studied in which the soil was used without fortification by either VC or IF. It was seen that up to 51.5%

greater germination success occurred in the VC treatments compared to controls. VC also supported better

plant growth in terms of stem diameter, shoot length, shoot mass, number of leaves, and leaf pigments. The

positive impact extended up to fruit yield. In addition, vermicast application enhanced root nodule formation,

reduced disease incidence, and allowed for a smaller number of stunted plants. The results indicate that

allelopathic ingredients of lantana seem to have been totally eliminated during the course of its

vermicomposting, and that lantana vermicompost has the potential to support germination, growth and fruit

yield better than equivalent quantities of inorganic fertilizers.

1. Introduction

Lantana (Lantana camara) is among the

world’s most noxious weeds (GISD, 2014). It is a

shrub, native to South America (Ghisalberti, 2000;

Henderson, 2001), and belongs to the family of

Verbenaceae which comprises of about 650 species

occurring in over 60 countries including India

(Sharma et al., 2005; Kohli et al., 2009; Hiremath

and Sundaram, 2013). Lantana grows in a wide

range of environmental conditions, infesting

millions of hectares of natural ecosystems and

cultivated lands, causing great damage to

biodiversity (Gentle and Duggin, 1997; Batianoff

and Franks, 1998; Day et al., 2003; Vardien et al.,

2012). It possesses mammalian toxicity and is

known to induce photodermatitis, jaundice, liver

damage and death in animals when they graze on its

leaves (Sharma et al., 1988, Pass, 1991; Bevilacqua

et al., 2011). It is also strongly allelopathic, having

several compounds which repel or toxify other

vegetation, thereby preventing their growth (Holm

et al., 1991; Gentle and Duggin, 1997; Day et al.,

2003; Ahmed et al., 2007).

Substantial efforts have been made, and are

continuing across the world, with billions of dollars

having been invested, to control invasives like

lantana by physical, chemical or biological means

(Erasmus and Clayton, 1992; McFadyen, 1998;

Day et al., 2003; Zalucki et al., 2007). However,

these attempts have not succeeded in even

82

controlling, let alone eradicating, any of the major

weeds. It appears that if processes can be developed

for gainfully utilizing the invasives, it may not only

offset the costs of mechanically removing them but

also exercise some control over their spread. One of

the possible options is conversion of weeds into

vermicompost and utilizing the latter as a soil

fertilizer.

Vermicomposting of a substrate is believed to

convert some of its nutrients into more bioavailable

forms and bestow upon the substrate microflora that

is beneficial for soil health (Gajalakshmi and

Abbasi, 2008; Edward et al., 2011). Additionally

hormones, enzymes and pest–repellants are

believed to be added to the substrate as it passes

through the earthworm gut and gets converted to

vermicast. Among these and several other attractive

features of the vermicomposting option is its ability

to sequester nearly all the carbon that is contained

in the substrate-to-be-vermicomposted (Abbasi et

al., 2011; Banupriya et al., 2014). But, despite the

great potential of vermicomposting as a phytowaste

utilization option, its use in handling phytomass in

general and weeds in particular has yet to begin

(Nayeem-Shah, 2014).

Recent reviews (Nayeem-Shah, 2014;

Nayeem- Shah et al., 2014) reveal that the main

constraint which has prevented the use of

vermicomposting in processing large quantities of

phytomass has been the incapability of the

conventional vermicomposting technology, which

is primarily geared to the utilization of animal

manure, in handling phytomass. To circumvent this

hurdle various authors have been, as reviewed by

Nayeem-Shah (2014), pre-composting the weeds

for 3 weeks or more, then subjecting the pre-

compost to vermicomposting after blending the

substrate with animal manure. Another 2-3 months

or more pass before the vermicomposting is

deemed to have completed. But the reliance on pre-

composting, animal manure, and the very long

overall processing time make the system

thoroughly unwieldly and uneconomical. The result

is that despite a spurt in laboratory- scale studies on

the vermicomposting of phytomass in recent years

(Nayeem-Shah, 2014), there is no attempt, nor it

appears possible, to utilize conventional

vermicomposting technology in processing

phytomass.

It has also been a matter of concern whether

phytomass-based vermicompost will be as

beneficial to soil and plants as manure-based

vermicompost is. This concern is particularly

relevant for plants like lantana which are known to

possess constituents that are toxic to animals and

other species of plants.

A solution to the first of the problems has

now emerged as a result of the efforts of S.A.

Abbasi and coworkers who have developed the

concept of high-rate vermicomposting and

associated technology (Gajalakshmi et al., 2002,

2005; Abbasi et al., 2009, 2011; Tauseef et al.,

2013). As detailed elsewhere (Nayeem-Shah et al.,

2013, 2014) the high-rate vermicomposting

technology makes it possible to directly

vermicompost phytomass; that, too, at a rapid rate.

Using this technology, it has also been possible to

directly vermicompost lantana and obtain its

vermicast in an efficient and convenient manner

(Kumar et al., 2012). With the prospect of

inexpensively and efficiently vermicomposting

lantana now assured, it has become imperative to

address the other major concern-the fertilizer value

of lantana vermicompost. The present paper is

devoted to this concern.


The studies were conducted at Pondicherry

University, Puducherry, India, located on the east

coast of the Indian peninsula (11°56’N, 79°53’E).

This region experiences hot summers during March

– July (maximum day temperature 35-38°C), and

mild winters during December - February

(maximum day temperature 29-32°C). The average

annual rainfall is about 1300 mm, concentrated

83

mainly during October – December but with a few

rainy days occurring in July–August and January as

well. For vermicomposting, reactors fabricated with

aluminum sheet of 140 liter volume were employed

for direct vermicomposting of lantana (Kumar et

al., 2012). The periodically harvested vermicast

was stored in sealed plastic containers. The study

on germination and growth was conducted outdoors

in 49 liter volume containers (40 cm height with

surface area 35 x 35 cm), lined with high-density

polyethylene (HDPE) sheets. The soil used in the

experiments was collected from a previously

uncultivated piece of land so that the results are not

influenced by any earlier fertilizer application. The

physico-chemical properties of the vermicast and of

the soil with which it was used are given in Table 1.

The experiments were carried out during July –

October which is ideal for growing cluster beans in

the study area (ICAR, 2011).

For studying the impact of vermicast on

cluster bean, three sets of experiments were

conducted. Based on the nitrogen, phosphorus, and

potassium (NPK) concentrations seen in the

vermicompost (Table 1), and the levels of NPK

needed in the soil for cluster bean (ICAR, 2011), it

was calculated that the cluster bean would require

7.5 t ha-1

of lantana vermicompost (VC) for its

optimum growth. In the first set, one batch of

containers was supplied with 7.5 t ha-1

of VC while

two other batches were given 5 and 10 t ha-1

of VC.

These doses were calculated on the basis of surface

area of the containers, and were 123, 92, and 61 g

for 10, 7.5 and 5 t ha-1

treatments, respectively.

Second set comprised of chemical fertilizers (IF)

which were treated with nutrients N, P, K, Ca, Mg,

S, Fe, Mn, Cu, Zn, B, Mo and Cl in concentrations

equivalent to those present in the respective

vermicompost treatments (Table 2). The nutrients

were supplied in two installments: half at the time

of sowing and the rest at the onset of flowering.

The last set had no supplementation of nutrients

and served as control. In this manner 3 sets of

experiments, encompassing seven treatments were

carried out. As each treatment had 36 containers,

each with one plant, 252 containers were utilized

for the main experiment. Another 150 containers,

30 of control and 20 each of the doses of VC and IF

treatments that were employed, were maintained as

spares. These were used to replace the containers of

which plants were sacrificed for analysis, or in

which the plants happened to have died. The

instances of plant death were very few, less than

3%. The Pusanavabahar variety cluster bean, which

is locally available, was used. Following the

assessment of germination over eight days, growth

and yield were monitored for an additional three

months. Throughout the experiments, adequate

watering was performed to maintain 20-30% (v/w)

moisture. Deweeding was done periodically. Neem

extract was applied as a mild pesticide in a few

instances when pests were seen infesting the plants.

2.1. Germination, plant growth and yield

characteristics

Seed germination was assessed for 8 days

from the day of sowing and was quantified in terms

of germination value (GV) using the formula of

Djavanshir and Pourbeik (1976):

Where GV is germination value; DGS is daily

germination speed which is computed by dividing

cumulative germination percent by the number of

days elapsed since the beginning of the test; GP is

germination percentage at the end of the test, and N

is frequency or number of DGS that are calculated

during the test. The unit for GV is germinated

seeds/day. It is used as an index to statistically

assess the effects of different treatments.

The growth was assessed on the basis of stem

diameter, length of shoot and root, number of

leaves, number of nodules present in the root, the

nodules size and shoot/root biomass for 3 months.

Plants were harvested periodically, and at each

harvest, the fruits were counted and their fresh and

84

Table 1. Chemical and physical properties of vermicast

and soil used in the study

BDL – Below detection limit

dry weights were determined. To quantify biomass

in terms of dry weight, samples were oven-dried at

85°C to constant weight. Chlorophyll a and b, and

carotenoids were extracted by the method described

by Moran and Porath (1980). One gram of mature

leaf was ground and incubated with N,N-di-methyl

formamide (DMF) for 24 h at 4 °C in dark, shaking

every 6 hours (Moran and Porath, 1980). The

resultant supernatant was read at 470, 647 and 664

nm and the concentration of pigments were

determined as detailed by Wellburn (1994). The

yield was calculated in the form of harvest index

(HI) which is the ratio of weight of beans per plant

to the above-ground biomass. The yield as reflected

in pod size, pea size, number of peas per pod and

diseased pods, if any, were recorded at each harvest

with randomly selected 50 fruits.

During the course of the experiment, the

plants were infected with bacterial blight

(Xanthomonas campestris) and alternaria blight

(Alternaria spp). Infestation of white fly (Bemisia

tabaci) also occurred. The bacterial blight infection

was seen in the symptoms of black leaf spots,

necrotic lesions at leaf tips, black streaks on

petioles and stems, split stems, and defoliation

(Mihail and Alcorn, 1985; Ren, 2014). Plants were

considered infected by Alternaria spp. when there

were dark brown lesions on leaves with concentric

zonations demarcated with light brown lines. In

severe infections, several spots merged and gave

the leaves a blighted look, eventually causing

defoliation (Orellana and Simmons, 1966;

Yogendra et al., 1995). The number of plants that

died, either with or without any symptoms of

infection, and stunted plants were also recorded.


Total organic carbon was measured by

modified dichromate redox method according to

Heanes (1984). Total nitrogen content was

determined by the modified Kjeldahl method


digester and distillation units. Inorganic N - NH4+

and NO3- were extracted in 2M KCl solution (1:10

weight: volume) and determined by modified

indophenol blue and Devarda’s alloy methods,

respectively (Jones, 2001; Bashour and Sayegh,

2007). Extractable potassium, calcium and sodium

were determined using a Flame photometer

(Elico™ CL378) after extraction with neutral 1N

ammonium acetate solution (Carter and Gregorich,

2008). Extractable magnesium, boron, copper, iron,

manganese, zinc, molybdenum were determined

using a Jobin Yvon – Ultima 2 model inductively

coupled plasma atomic emission spectroscopy (ICP

Variables Concentration

Vermicast Soil

Chemical properties

pH 6.47±0.01 6.30±0.10

Total organic carbon g kg-1

330.3±4.3 8.87±0.02

Total Nitrogen g kg-1

18.4±0.1 2.66±0.02

Plant available form of

Phosphorus mg kg -1

80.7±0.9 0.41±0.01

Potassium g kg-1

5.73±0.03 0.40±0.00

Sulphur mg 100g-1

1.34±0.01 0.54±0.01

Calcium g kg-1

5.49±0.02 8.27±0.01

Magnesium g kg-1

5.77±0.05 0.09±0.02

Boron mg kg-1

46.2±0.8 26.9±1.2

Copper mg kg-1

44.2±3.9 5.08±0.15

Iron mg kg-1

50.6±2.0 59.9±2.9

Manganese mg kg-1

227.2±8.4 45.1±2.2

Zinc mg kg-1

162.4±6.0 55.0±2.6

Molybdenum mg kg-1

BDL BDL

Physical properties

Dry weight % 43.1±0.3 94.7±0.1

Bulk density g cm-3

0.40±0.00 1.28±0.00

Particle density g cm-3

1.35±0.01 2.70±0.12

Water-holding capacity % 248.0±10.2 36.9±2.4

Electrical conductivity

mmhos cm-1

9.36±0.01 0.12±0.02

Total porosity % 70.6±0.2 53.1±2.0

Air filled porosity % 48.1±0.1 46.5±2.1

85

Table 2. Amount of inorganic fertilizer applied equivalent to vermicast treatment

Nutrients Form in which

applied

Mass %

of nutrient

in applied

compound

10t ha-1

7.5t ha-1

5t ha-1

Concentra

tion of

nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Concentration

of nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Concentration

of nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Nitrogen CH4N2O 46.65 66.26 142.1a 49.70 106.5

a 33.13 71.03

a

Phosphorus (NH4)2HPO4 23.45 34.79 148.3 26.09 111.3 17.40 74.17

Potassium KCl 52.44 24.82 47.32 18.61 35.492 12.41 23.66

Sulphur 0.058 Nilb 0.043 Nil

b 0.029 Nil

b

Calcium CaCO3 40.04 23.70 59.19 17.78 44.39 11.85 29.59

Magnesium MgO 60.30 24.91 41.31 18.68 30.98 12.45 20.65

Boron Na2B4O7.10H2O 11.34 0.199 1.756 0.149 1.317 0.100 0.878

Copper CuSO4.5H2O 25.45 0.191 0.749 0.143 0.562 0.095 0.375

Iron FeSO4.7H2O 20.09 0.218 1.086 0.164 0.815 0.109 0.543

Manganese MnSO4. H2O 32.50 0.980 3.015 0.735 2.262 0.490 1.508

Zinc ZnCl2 47.97 0.700 1.460 0.525 1.095 0.350 0.730 a The total amount of N applied in the form of urea and di-ammonium phosphate is equal to the concentration of N applied in the

vermicast treatment. b The total amount of sulphate applied with other elements equalizes the sulphate in applied vermicast.

– AES) by extracting sample/solution ratio of 1:25

with Mehlich 3 extraction solution (Mehlich, 1984).

The same extract was used to determine the

extractable phosphorus according to the ammonium

molybdate-ascorbic acid method (Knudsen and

Beegle, 1988). Mineral S in soil was extracted with

0.0125M CaCl2 solution (ratio of soil: solution,

1:4), and analyzed with a turbidimeter after the

addition of BaCl2 and generating BaSO4 turbidity

(Bashour and Sayegh, 2007).

The pH and electrical conductivity (EC) of

the samples were measured in suspension of 1:2

(v/w) by using EI™ 611E EC meter and Digison™

digital pH meter 7007 respectively. Bulk density

was measured on undisturbed cores for soil and the

graduated cylinder method was used for vermicast.

Particle density was determined by volumetric flask

method (Bashour and Sayegh, 2007). The total and

water-filled porosity were calculated from the

particle and bulk density values of respective

samples using standard formulae (Carter and

Gregorich, 2008). Water holding capacity (WHC)

of the samples was obtained by determining their

water retention ability after they were immersed in

water and the excess water was drained off

(Margesin and Schinner, 2005).

2.4. Statistical analysis

One-way ANOVA and post hoc LSD tests

(Weinberg and Abramowitz, 2008) were employed

for determining significance of difference between

the results (SPSS version 16; Softonic, Barcelona,

Spain).


3.1. Seed germination

The GV and GP were significantly higher in

the VC and IF treatments compared to the control

(F2,32 = 3.759, p <0.05; Table 3). The VC

application at the dose of 5 t ha-1

increased the GP

by 8.3% compared to the control. Further increase

in the dose of VC caused an increase in GP to

51.5% higher than the control. In the IF treatments

too, the GP significantly improved with increasing

86

Table 3. Germination value (GV) and germination percentage (GP) of the seeds of cluster bean as influenced

by lantana vermicast and inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05)

carry at least one character in the superscript which is common.

Treatment Amount Fourth day Fifth day Sixth day Seventh day Eighth day

GV GP GV GP GV GP GV GP GV GP

Vermicompost

5t ha-1 a

3.27 22.86 2.64 25.71 5.10 42.86 4.54 47.14 4.13 51.43

7.5 t ha-1 bc

37.19 77.14 23.80 77.14 17.78 80.00 14.01 82.86 10.73 82.86

10 t ha-1 b

55.57 94.29 37.74 97.14 26.21 97.14 19.26 97.14 14.74 97.14

Inorganic

fertilizer

equivalent to

the VC

5t ha-1 acd

26.99 65.71 17.27 65.71 13.06 68.57 10.41 71.43 7.97 71.43

7.5 t ha-1 bd

34.49 74.29 23.80 77.14 19.07 82.86 14.01 82.86 10.73 82.86

10 t ha-1 b

37.19 77.14 25.60 80.00 19.07 82.86 14.01 82.86 11.48 85.71

Control Nil a 2.50 20.00 2.64 25.71 2.74 31.43 4.26 45.71 3.47 47.14

dosage, and up to 45% higher GP than in control

was recorded at the dose of 10 t ha-1

. Except VC

treatment at 5 t ha-1

, in all other treatments,

maximum GV was observed on the 4th day

followed by a steady decline. In 5 t ha-1

VC, the

maximum GV was recorded on the 6th day. All-in-

all, the maximum GV of 55.6 was recorded with

VC treatment at 10 t ha-1

, followed by 10 t ha-1

of

IF and 7.5 t ha-1

of VC treatments, respectively,

which both had a GV of 37.2.

It is possible that the GP increased with

increased doses of VC and IF as a result of

increasing concentration of nitrate and ammonium

contained in them. As these compounds break

dormancy and stimulate germination (Egley and

Duke, 1985; Hilhorst and Karssen, 2000),

increasing the concentration of VC and IF would

have increased the germination success. In general,

germination is stimulated within a range of 0–0.05

M nitrate and to a certain extent, ammonium ion

also influences the seed germination (Hilhorst and

Karssen, 2000). In the present study, 0.9 to 1.8 g of

nitrate was supplemented in the form of either VC

or IF, which would have increased the germination

in the initial days. Since the nitrate readily leaches

from surface soil by irrigation water, the

concentration of nitrate might have declined as the

days progressed, causing a reduction in germination

success. Moreover, since greater germination

success was recorded with VC treatment than

equivalent IF treatments, phytohormones like

gibberellins and other organic chemicals which are

known to enhance seed germination might have

been involved (Miransari and Smith, 2014) to make

VC more effective than IF.

3.2. Plant growth

VC was seen to exert much more favorable

impact on plant growth than IF, as reflected in all

the plant growth parameters that have been studied

(Table 4). The plants which were treated with VC

showed higher stem diameter, length, fresh

weight/dry weight of shoot, and number of leaves

than plants treated with equivalent IF, or the

control. The beneficial impact of VC may be due to

a number of reasons, including favorable nodules

formation, nutrient release synchronized with plant

need, and the contribution of beneficial plant

growth regulators, hormones and microbes. More

nodules were observed with the plants which were

treated with VC than with IF, and greater

nodulation was witnessed at higher dosage of VC.

This finding is highly significant because

nodulation is a process which is tightly controlled

in nature to allow only as much symbiont rhizobia

in the roots of a legume as necessary for the legume

(Mortier et al., 2012). Several feedback routes exist

in nature to accomplish this control (Miwa et al.,

87

Table 4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth /death in cluster bean plants as impacted by lantana

vermicast or inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05) carry at least one character in the superscript which is

common.

*p<0.05,

**p<0.01,

***p<0.001,


Parameters

Vermicompost at dose Inorganic fertilizers at dose

Control

F-value

5 t ha-1

7.5 t ha-1

10 t ha-1

5 t ha-1

7.5 t ha-1

10 t ha-1

Type of

fertilizer Amount

Plant growth

Stem diameter (mm) 13.6±0.7a 14.9±1.2

b 17.2±1.2

c 12.9±1.0

a 13.8±1.4

a 17.0±1.5

c 11.2±1.7

d 18.45

*** 63.69

***

Shoot length (cm) 168±7.0a 203.0±12.6

b 269.7±24.4

c 175.8±12.1

a 195.1±11.8

b 215.4±11.7

d 76.8±5.9

e 80.46

*** 189.0

***

Root length (cm) 74±9.3a 68.2±8.7

ab 65.7±6.9

bc 68.4±5.5

acd 64.9±7.2

bd 52.7±4.5

e 50.2±8.4

e 22.09

*** 20.56

***

Number of leaves 83.6±4.6a 96±5.8

b 99.4±7.2

b 79.3±5.2

ac 81.9±5.3

a 86.5±6.4

ad 42.0±5.3

e 211.2

*** 128.2

***

Number of nodules 62.2±11.3a 69.6±15.2

a 70.7±16.6

a 39.1±7.4

b 35.9±9.9

b 36.1±10.5

b 13.5±6.0

c 101.5

*** 16.03

***

Shoot dry weight (g) 54.2±6.9a 69.3±7.8

b 108.5±13.1

c 19.8±4.7

d 71.5±9.5

b 80.7±11.6

e 45.3±5.2

f 9.690

*** 67.34

***

Root dry weight(g) 5.9±1.0ad

6.5±1.2a 7.7±1.1

b 4.5±1.0

c 5.5±0.9

de 5.1±0.8

ce 3.2±0.6

f 50.18

*** 18.65

***

Leaf pigments

Chlorophyll a (mg g-1

) 2.41±0.06a 3.24±0.13

b 3.79±0.18

c 2.59±0.02

d 2.92±0.05

e 2.95±0.03

e 1.32±0.05

f 92.73

*** 174.4

***

Chlorophyll b (mg g-1

) 2.23±0.06a 2.49±0.07

b 2.88±0.08

c 2.39±0.04

d 2.66±0.06

e 2.72±0.03

f 1.45±0.04

g 143.1

*** 573.7

***

Total chlorophyll (mg g-1

) 4.64±0.07a 5.73±0.13

b 6.68±0.21

c 4.97±0.04

d 5.58±0.07

e 5.66±0.04

b 2.77±0.04

f 111.5

*** 348.5

***

Carotenoids (mg g-1

) 0.49±0.05a 0.57±0.03

b 0.59±0.02

b 0.51±0.02

ac 0.52±0.02

c 0.36±0.02

d 0.26±0.02

e 94.50

*** 47.47

***

Flowering

Number of flowers 7.2±4.7a 14.7±5.0

b 25.4±6.8

c 7.9±3.7

a 16.2±6.7

bd 18.4±8.3

d 0.7±1.0

e 49.96

*** 131.2

***

Disease incident, plant death and stunted plants

Number of infected plant 14a 5

a 9

a 18

a 10

a 11

a 16

a 0.735

n.s 1.228

n.s

Number of plant died Nila Nil

a 2

a Nil

a 1

a 2

b Nil

a 0.402

n.s 2.022

n.s

Number of stunted plant Nila 1

a Nil

a 1

a 12

a 5

a 11

a 1.824

n.s 1.026

n.s

88

2006; Miyazawa et al., 2010). Evidently, VC

generates signals that favour nodule formation. In

contrast IF treatments may induce depression in

nodulation because they provide the nitrogen

contained in the IF all at once leading to signals

that discourage nodulation (Carroll and Gresshoff,

1983; Harper and Gibson, 1984; Nie, 1989). van

Schreven (1959) had reported suppression of

growth in clover (Trifolium repens) and lucerne

(Medicago sativa) nodules when urea was applied

at concentration as low as 0.5%. Presence of higher

Mn in the IF may also have contributed to the

suppression of nodulation in the IF treatment

(Dobereiner, 1966; Foy et al., 1978). Even though

VC also contains the same inorganic nutrients, and

in the same concentration, as IF, the release from

VC might have been at much slower rate because of

the predominantly organic matrix of the VC. This

might have made nutrients available to the legume

at a rate best suited to promote nodules formation.

The slow release of nutrients by the VC may

also have caused lesser nutrient loss from the

rhizosphere. The presence of biologically active

substances such as fulvic acids, humic acids and

phytohormones in the VC may have contributed to

better plant growth in VC treated plants.

Specifically, cytokinins have been reported in

vermicast (Zhang et al., 2014). It has also been

shown that humic acids derived from the vermicast

induce morphogenetic and biological changes

favorable to plants which are similar to those

produced by indole-3-acetic acid (Muscolo et al.,

1999). These bioactive substances are probably

produced due to the abundance of microbial

communities in the vermicompost, specifically the

actinomycetes and fungal species, which then

releases phytohormones in the soil (Frankenberger

and Arshad, 1995). Improvement in the physical

properties of the soil, such as aeration and water

holding capacity, may have also contributed to an

increase in the plant productivity in soil treated

with VC (Edwards, 2004).

3.3. Photosynthetic pigments

Photosynthetic pigments chlorophyll and

carotenoid in the leaves of plants were significantly

influenced by the VC and IF application at different

doses (Table 4). The pigment levels were seen to

increase with increasing rate of fertilizer

application as there was concomitant increase in the

nutrient availability (Mengel and Kirkby, 1987;

Shadchina and Dmitrieva, 1995; Ruza, 1996;

Tejada et al., 2007). Higher concentrations of the

pigments were recorded in the VC treatments of 7.5

and 10 t ha-1

than in the equivalent IF treatments.

Only in VC treatment at 5 t ha-1

lesser amount of

chlorophyll a and b was found than in the

equivalent inorganic fertilizer treatment. The

carotenoid content was also higher in the VC

treated plants than the IF treated ones. This also

confirms that VC application increased the nutrient

availability in soil and enhanced nutrient uptake by

plants in comparison to the IF treatment.

3.4. Flowering

Plants grown in soil treated with VC had

significantly larger number of flowers than plants

grown in IF- treated soil (p<0.001; Table 4).

Greater the fertilizer application, more profuse was

the flowering; the trend with both VC and IF being

10 t ha-1

> 7.5 t ha-1

> 5 t ha-1

> control.

3.5. Disease incidence, plant death and stunted

growth

During the first two months of the

experiments, the plants were found to be infected

with bacterial blight (Xanthomonas campestris),

and alternaria blight (Alternaria spp) and infested

with whiteflies (Bemisia tabaci) (Table 4). The

maximum infestation was seen in the 5 t ha-1

IF

treatments followed by control and 5 t ha-1

VC

treatments. Lesser number of infected plants were

observed in VC fertilized plants than in the plants

that were given equivalent IF treatments. Previous

89

Table 5. Harvest index and yield attributes of plants as impacted by lantana vermicast and equivalent

inorganic fertilizers (mean ±SD). Results which do not differ significantly (LSD test; p <0.05) carry at least

one character in the superscript which is common.

*p<0.05, **p<0.01, ***p<0.001,n.s - not significant.

studies on vermicast obtained from animal manure

have shown that the vermicast possesses pest–

repellant properties (Edwards et al., 2011). The

present studies indicate that the lantana vermicast

may also be imbibed with a similar attribute. The

better nutrient availability made possible by the VC

might also be contributing to the development of

resistance to pathogens in the plants because

availability of adequate nutrients enhances the

ability of the plants to limit the penetration,

development, and/or reproduction of invading

pathogens (Graham and Webb, 1991). Some of the

nutrients are also involved in the production of

antimicrobial compounds such as flavonoids and

phenolics that act against the plant pathogens

(Graham and Webb, 1991; Hill et al., 1999). The

humic acid content of the vermicast is also likely to

affect biochemical processes in the plants and

bacteria, resulting in induction of resistance in

plants to certain phytopathogens (Sahni et al.,

2008).

Moreover, lesser number of VC treated plants

exhibited stunted growth than the IF treated ones

and the control (Table 4). A few plants died during

the experiment, but there was no significant

difference in the number among different

treatments (Table 4).

3.5. Fruit yield

The number and weight of pods per plant

were significantly higher in VC treatments than the

equivalent IF treatments (p<0.05) (Table 5).

Among all the treatments, highest yield of pods was

recorded with VC treatment at 7.5 t ha-1

followed

by the equivalent IF treatment. The weight and the

number of fruits harvested from 7.5 t ha-1

VC

treated plants was about one fold greater than the

corresponding IF treatment. At higher fertilizer

dose (10 t ha-1

) this beneficial effect was reversed.

It is known that application of nutrients which is

well above the levels required by a plant may

induce higher immobilization of nutrients in the

plant biomass and less partitioning to fruits (Wada

et al., 1989; Sujatha and Bhat, 2013). A similar

factor might have been behind the reduction of

yield in the 10 t ha-1

VC/IF applications.

Table 5 shows that the harvest index (HI)

values of VC treated plants are not significantly

higher than that of the IF treated plants even as the

former have given significantly higher fruit yield.

This is because HI is a ratio of mass of fruits and

mass of the above-ground biomass. In VC treated

plants both are higher than in IF treated plants

leading to similar HI.

Parameters, average

value

Vermicompost at dose Inorganic fertilizer at dose F-value

5 t ha-1 7.5 t ha-1 10 t ha-1 5 t ha-1 7.5 t ha-1 10 t ha-1 Type of

fertilizer Amount

No.of pod per plant 18.7±6.6a 40.0±8.2c 29.9±7.2b 14.1±5.9d 25.5±6.3b 20.5±7.1a 119.1*** 95.50***

Weight of the pods (g) 93.5±29.0a 227.0±45.0b 162.5±39.5c 77.9±31.9d 145.4±37.4e 110.5±36.5a 139.0*** 154.8***

Pod length (cm) 15.1±1.7a 16.0±1.4b 16.0±1.6b 14.8±1.8a 15.0±1.8a 14.7±1.8a 18.63*** 2.744n.s

Pod width (cm) 10.2±1.6a 11.3±1.5b 10.6±0.8ac 10.6±1.4ad 11.0±1.4bcd 10.8±1.9bcd 0.385n.s 6.249**

Pod thickness (cm) 6.19±0.84aeg 6.66±0.79b 6.35±0.5ab 5.87±1.02adg 6.02±0.71cdef 5.87±0.62fg 24.18*** 3.471*

No.of seeds per pod 9.20±1.25ab 9.56±0.97a 9.42±1.20ac 9.02±0.94bc 9.48±1.09a 9.46±0.95a 0.368n.s 4.116*

Seed diameter (mm) 5.60±0.86a 6.14±0.72b 6.47±0.73c 5.59±0.97a 6.00±0.76d 6.52±0.71c 0.917n.s 294.7***

Seed thickness (mm) 2.24±0.47a 2.49±0.45b 2.54±0.55bc 2.07±0.63d 2.37±0.55e 2.55±0.47c 17.13*** 132.8***

Harvest Index % 69.8±40.1abcde 107.0±71.2be 50.4±37.1c 74.8±50.8abcde 91.9±78.8ab 51.3±34.8cde 0.965n.s 5.107*

90

The length, width and thickness of the pods,

and the average number of seeds per pod, were

higher in plants treated with VC than in the

equivalent IF treatment. The VC and IF treatments

at 7.5 t ha-1

showed these yield attributes in better

measure than the 5 and 10 t ha-1

treatments,

whereas seed diameter and seed thickness were

maximum in 10 t ha-1

treatments.

3.6. The present study in the context of the state-

of-the-art

There is only one pre-existing report on this

subject, that of Suthar and Sharma (2013). They

had made four blends of lantana and cow manure

(CM) in 1:4, 1:1.5, 1.5:1, and 4:1 mass ratios,

composted it for 3 weeks, then kept 250 g dry

weight equivalent of each pre-composted blend

with 10 individuals of Eisenia fetida for 60 days.

They then found that the resulting worm-worked

reactor content supported germination of the seeds

of corn (Zea mays). It is difficult to conclude much

from this study because each treatment had so

much CM in it that the very low number of

earthworms, 10, that were used in each treatment

might as well have ingested CM particles for most

part. Secondly, once lantana or any other

phytomass is brought in contact with CM, the

cellulolytic and acidogenic bacteria contained in the

CM begin biodegrading the phytomass to produce

volatile fatty acids (VFAs; Kumar et al., 2014).

Suthar and Sharma (2013) have reported that there

was a sharp fall in pH in their treatments which also

indicates that VFA formation might have occurred.

This is likely to have effected the entire chemistry

and microbiology of the treatments and much of the

data reported by Suthar and Sharma (2013) is likely

to have been influenced by this happening. In

particular, lowering of pH is known to make trace

nutrients more labile which might have contributed,

rather than vermicomposting, in the beneficial

effects on the germination of corn as seen by them.

Moreover, unless vermicomposting is defined and

quantified in terms of vermicast deposited by the

earthworm―which is always an easily

distinguishable, separable, and quantifiable entity in

any vermireactor (Tauseef et al., 2014) ― it is

never possible to design and optimize a

vermireactor. The worm–worked substrate, as

deemed vermicompost by Suthar and Sharma

(2013) and other authors before them (Nayeem-

Shah 2014), is a non-descript entity, being a

mixture of vermicast, partially biodegraded

substrate, and numerous products of natural and

worm-mediated biodegradation occurring

uncontrolled in such vermireactors, of which exact

proportions or even nature is very difficult to

quantify. Nor based on such a parameter, it can be

said when the vermireactor attains steady state or

when the vermicomposting is complete. As

vermicast is not periodically removed from these

reactors, but is instead periodically homogenized

with the rest of the reactor content, the earthworm

may be reingesting much of it, thereby harming the

reactor efficiency while the uncontrolled

biodegradation of the rest of the unharvested

vermicast would make process monitoring or

control even more difficult.

Due to all these reasons, it is difficult to draw

any meaningful conclusion from the study of Suthar

and Sharma (2013) on either vermicomposting of

lantana or its fertilizer value. In contrast, the

present report is based on lantana vermicompost

which was generated by direct vermicomposting of

lantana with Eudrilus eugeniae in pulse-fed, high-

rate, vermireactors by a process detailed elsewhere

(Kumar et al., 2012). The vermicast, which was

periodically harvested in each pulse, was obtained

as a clear and precisely quantifiable product of the

vermireactors and was termed vermicompost. The

scientific basis of this definition has been given

earlier (Abbasi et al., 2009), and has been included

as a valid definition in Edward et al. (2011). In

view of this it can be said that the present report

shows rather conclusively that vermicompost

derived from phytomass can be as plant-friendly as

manure-based vermicompost is known to be, even

when it is derived from an allelopathic weed like

91

lantana which, additionally, possesses mammalian

toxicity.

4. Conclusions

The paper describes results of a study, the

first of its kind, in which the effect of

vermicompost (VC) derived solely from an

allelopathic weed has been studied on the

germination, growth and fruition of a botanical

species. The impact of the VC was compared with

that of an inorganic fertilizer (IF) which had all the

main macro and micro – nutrients in concentrations

equivalent to the ones present in the VC. Several

sets of experiments were carried out in which the

soil was treated with either VC obtained from

lantana (Lantana camara) or the IF at the rates of 5,

7.5 or 10 tons per hectare (t ha-1

). Cluster bean

(Cyamopsis tetragonoloba) was the botanical

species used in the study; chosen because it is a

common vegetable and a legume. Additionally a set

of controls was studied in which the soil was used

without fortification by either VC or IF.

It was seen that significantly greater

germination rates occurred in the VC treatments

compared to controls. VC also supported better

plant growth in terms of stem diameter, shoot

length, shoot mass, number of leaves, and leaf

pigments. The positive impact extended up to pod

yield. In addition, vermicast application enhanced

root nodule formation, reduced disease incidence,

and permitted lesser number of stunted plants.

The findings reveal that the allelopathic

ingredients of lantana seem to have been totally

eliminated during the course of its

vermicomposting, and that lantana vermicompost

has the potential to support germination, growth

and fruit yield better than equivalent quantity of

inorganic fertilizers. The study opens up the

possibility that other allelopathic weeds, as also

plants which are toxic in other ways, may be

utilizable as substrates in high-rate vermireactors as

vermicomposting is likely to destroy the toxic

components of these substrates as it is seen to have

done in case of lantana. This, in turn, may

enormously enhance the applicability of

vermicomposting as well as provide a means of

utilizing the biomass of several invasives which,

otherwise, goes to waste.

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Supplementary material

Table 4A. Stem diameter (mm) of cluster bean at different weeks

Treatment Amount Weeks

1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

1.1 1.4 2.1 3.4 4.2 4.9 7.6 8.2 10.3 11.2 13.1 13.6

7.5 t ha-1

1.6 1.7 2.4 3.5 4.6 8.1 8.2 8.4 11.1 12.5 13.2 14.9

10 t ha-1

1.7 2.0 2.2 4.1 6.6 10.0 10.7 11.5 13.6 14.8 15.5 17.2

Inorganic

fertilizer

5 t ha-1

1.3 1.9 2.2 3.7 4.6 5.2 6.2 7.3 8.7 9.8 11.9 12.9

7.5 t ha-1

1.6 2.0 2.1 2.5 4.6 7.3 7.6 8.9 9.4 12.2 12.4 13.8

10 t ha-1

1.7 2.0 2.1 3.2 4.7 6.7 8.3 8.8 10.2 12.4 13.4 17.0

Control Nil 0.7 1.8 2.0 2.3 4.5 5.2 5.8 6.1 1.2 10.8 11.1 11.2

Table 4B. Shoot length (cm) of cluster bean at different weeks


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

9.8 13.4 15.6 17.1 27.8 42.2 68.4 75.7 91.2 105 115 168

7.5 t ha-1

9.9 11.5 16.1 20.5 36.5 54.9 74.5 106.0 115.8 138 166 203

10 t ha-1

11.5 13.5 18.9 21.0 42.8 62.2 111.5 129.5 148.0 194 220 270

Inorganic

fertilizer

5 t ha-1

10.1 14.1 17.5 18.9 28.4 45.8 70.6 78.2 95.7 117 129 176

7.5 t ha-1

10.5 12.3 18.7 19.9 29.7 49.2 76.7 97.0 101.3 119 152 195

10 t ha-1

10.9 13.3 18.4 20.1 30.0 52.5 83.2 103.5 105.5 129 148 215

Control Nil 3.5 11.4 11.8 12.5 18.0 21.8 23.0 26.0 34.5 42.0 59.3 76.8

Table 4C. Root length (cm) of cluster bean at different weeks


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

6.3 13.9 22.0 26.9 28.5 38.6 52.5 63.5 65.4 70.2 72.1 74.0

7.5 t ha-1

6.7 19.4 25.3 21.0 26.6 37.6 49.6 62.0 63.4 64.0 66.6 68.2

10 t ha-1

8.9 17.1 20.0 20.5 24.5 37.0 44.0 48.7 56.3 62.1 63.2 65.7

Inorganic

fertilizer

5 t ha-1

6.7 19.4 21.0 28.5 29.3 31.6 40.5 58.7 59.2 61.7 63.6 68.4

7.5 t ha-1

8.2 19.8 20.1 32.4 27.9 29.6 38.5 56.9 58,3 60.2 61.3 64.9

10 t ha-1

8.7 20.5 20.8 21.0 25.2 28.3 37.9 47.6 51.3 53.9 56.4 52.7

Control Nil 3.8 13.7 14.0 23.1 29.1 29.2 37.4 42.0 45.7 47.5 49.3 50.2

97

Table 4D. Number of leaves of cluster bean at different weeks


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

03 03 04 9 18 28 34 42 60 69 72 84

7.5 t ha-1

03 04 06 12 22 36 45 52 58 70 78 96

10 t ha-1

03 04 06 13 22 37 49 55 63 81 88 99

Inorganic

fertilizer

5 t ha-1

03 03 04 10 19 26 32 38 58 59 63 79

7.5 t ha-1

03 04 07 11 20 30 34 46 62 63 72 82

10 t ha-1

03 04 08 12 21 32 38 50 64 68 72 87

Control Nil 03 03 04 09 15 17 21 25 28 31 35 42

Table 4E. Number of nodules of cluster bean at different weeks


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

Nil 11 12 15 11 22 34 38 33 52 60 62

7.5 t ha-1

Nil 14 15 24 18 32 40 48 60 52 66 70

10 t ha-1

Nil 15 14 15 22 27 36 65 45 115 82 71

Inorganic

fertilizer

5 t ha-1

Nil 10 14 12 15 6 37 25 12 19 31 39

7.5 t ha-1

Nil 3 7 6 10 22 12 46 26 47 17 36

10 t ha-1

6 17 16 10 14 24 16 38 41 22 30 36

Control Nil Nil 0 7 8 12 14 19 11 11 9 16 14

Table 4F. Shoot dry weight (g) of cluster bean at different weeks


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.033 0.210 0.295 0.551 0.967 4.914 7.990 13.80 25.14 36.25 48.29 54.17

7.5 t ha-1

0.076 0.310 0.486 0.738 2.971 7.312 12.30 20.13 34.72 52.05 60.48 69.27

10 t ha-1

0.090 0.346 0.601 1.080 4.252 11.36 28.50 35.14 54.63 68.53 87.82 108.5

Inorganic

fertilizer

5 t ha-1

0.055 0.211 0.354 0.359 1.169 3.871 6.184 11.30 15.82 23.61 27.56 19.82

7.5 t ha-1

0.128 0.247 0.322 0.429 1.359 4.570 10.11 14.74 21.86 41.71 47.49 71.54

10 t ha-1

0.102 0.304 0.464 0.470 1.465 6.784 16.58 21.50 29.27 46.74 60.48 80.66

Control Nil 0.032 0.109 0.205 0.395 0.955 1.490 4.572 6.799 12.11 41.50 44.08 45.32

98

Table 4G. Root dry weight (g) of cluster bean at different weeks

Table 4H. Leaf chlorophyll a (mg g-1

) content of cluster bean at different weeks


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.359 1.475 1.521 1.899 1.059 2.131 2.197 2.238 2.297 2.316 2.395 2.407

7.5 t ha-1

0.305 1.477 1.734 1.967 2.291 2.486 2.781 2.931 3.062 3.155 3.197 3.240

10 t ha-1

0.563 1.954 2.192 2.802 2.826 3.169 3.447 3.501 3.587 3.626 3.684 3.793

Inorganic

fertilizer

5 t ha-1

0.581 1.24 1.597 1.88 2.069 2.243 2.331 2.402 2.436 2.491 2.538 2.586

7.5 t ha-1

0.491 1.604 1.934 2.159 2.214 2.356 2.555 2.725 2.762 2.834 2.909 2.916

10 t ha-1

0.323 1.181 1.925 2.175 2.253 2.44 2.755 2.792 2.852 2.873 2.922 2.945

Control Nil 0.103 0.465 0.64 0.907 1.128 1.164 1.177 1.193 1.201 1.224 1.238 1.316

Table 4I Leaf chlorophyll b (mg g-1



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.206 0.568 1.173 1.258 1.306 1.584 1.789 2.058 2.126 2.170 2.201 2.230

7.5 t ha-1

0.262 0.603 0.874 1.151 1.363 1.591 1.822 2.316 2.394 2.446 2.482 2.491

10 t ha-1

0.241 0.587 1.271 1.421 1.782 1.975 2.395 2.550 2.673 2.719 2.783 2.884

Inorganic

fertilizer

5 t ha-1

0.193 0.565 0.938 0.976 1.129 1.779 1.947 2.198 2.228 2.297 2.342 2.385

7.5 t ha-1

0.261 0.511 1.212 1.413 1.747 1.867 2.193 2.435 2.512 2.579 2.614 2.663

10 t ha-1

0.358 0.552 1.249 1.593 1.812 1.909 2.246 2.466 2.511 2.579 2.652 2.719

Control Nil 0.032 0.155 0.223 0.504 1.612 1.935 1.065 1.409 1.510 1.532 1.544 1.451


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.013 0.021 0.051 0.071 0.184 0.514 0.680 2.403 2.922 3.829 4.933 5.925

7.5 t ha-1

0.010 0.022 0.059 0.087 0.343 0.761 0.982 2.782 3.360 4.259 5.343 6.511

10 t ha-1

0.013 0.028 0.106 0.113 0.421 0.901 1.200 2.913 3.531 4.680 5.689 7.680

Inorganic

fertilizer

5 t ha-1

0.009 0.031 0.036 0.054 0.179 0.435 0.520 1.771 2.660 3.736 4.472 4.532

7.5 t ha-1

0.011 0.044 0.054 0.068 0.202 0.530 0.832 2.316 3.083 4.143 4.905 5.539

10 t ha-1

0.009 0.053 0.083 0.105 0.347 0.658 0.999 2.585 3.397 3.984 5.000 5.121

Control Nil 0.001 0.014 0.016 0.029 0.102 0.256 0.289 0.981 1.763 2.929 3.160 3.214

99

Table 4J. Leaf total chlorophyll (mg g-1



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.565 2.043 2.694 3.157 2.365 3.715 3.986 4.296 4.423 4.486 4.596 4.637

7.5 t ha-1

0.567 2.08 2.608 3.118 3.654 4.077 4.603 5.247 5.456 5.601 5.679 5.731

10 t ha-1

0.804 2.541 3.463 4.223 4.608 5.144 5.842 6.051 6.260 6.345 6.467 6.677

Inorganic

fertilizer

5 t ha-1

0.774 1.805 2.535 2.856 3.198 4.022 4.278 4.600 4.664 4.788 4.880 4.971

7.5 t ha-1

0.752 2.115 3.146 3.572 3.961 4.223 4.748 5.16 5.274 5.413 5.523 5.579

10 t ha-1

0.681 1.733 3.174 3.768 4.065 4.349 5.001 5.258 5.363 5.452 5.574 5.664

Control Nil 0.135 0.62 0.863 1.411 2.740 3.099 2.242 2.602 2.711 2.756 2.782 2.767

Table 4K. Leaf carotenoids (mg g-1



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.138 0.179 0.215 0.253 0.299 0.311 0.32 0.376 0.384 0.414 0.426 0.490

7.5 t ha-1

0.151 0.276 0.352 0.381 0.416 0.428 0.466 0.481 0.499 0.512 0.535 0.572

10 t ha-1

0.116 0.285 0.394 0.421 0.441 0.467 0.475 0.493 0.506 0.538 0.562 0.589

Inorganic

fertilizer

5 t ha-1

0.125 0.206 0.319 0.358 0.395 0.418 0.434 0.457 0.465 0.482 0.491 0.511

7.5 t ha-1

0.154 0.255 0.335 0.379 0.407 0.417 0.427 0.456 0.471 0.489 0.506 0.524

10 t ha-1

0.151 0.183 0.214 0.237 0.252 0.285 0.312 0.314 0.318 0.337 0.341 0.359

Control Nil 0.102 0.122 0.136 0.158 0.164 0.191 0.203 0.218 0.224 0.236 0.248 0.261

EFFECT OF VERMICAST GENERATED FROM A

PERNICIOUS WEED IPOMOEA (IPOMOEA CARNEA)

ON SEED GERMINATION, PLANT GROWTH, AND

YIELD OF CLUSTER BEAN (CYAMOPSIS

TETRAGONOLOBA)

Chapter

7

101



CChhaapptteerr 77

Effect of vermicast generated from a pernicious weed ipomoea

(Ipomoea carnea) on seed germination, plant growth, and yield

of cluster bean (Cyamopsis tetragonoloba)

Abstract

The impact of vermicast derived from the highly pernicious, amphibian weed, Ipomoea carnea on cluster

bean (Cyamopsis tetragonoloba) was assessed in terms of germination, growth and fruit yield. Seeds of

cluster bean were sown in soils to which vermicast was applied at the rates of 5, 7.5 and 10 t ha-1

. The impact

of vermicast on seed germination, growth and yield of experimental plants were compared with the plants

which were treated with inorganic fertilizers (IF) in concentrations equivalent to those present in the

respective vermicompost (VC) treatments. Additionally a set of control was studied in which the soil was

used without fortification by either vermicompost or inorganic fertilizers. Following assessment of

germination rates, the plant growth parameters were determined each week for 3 months with randomly

collected samples. The yield attributes were assessed with manually harvested mature pods from all the

experimental plants. The effect of different treatments on germination, growth and yield attributes are very

significant. The vermicast treated plants showed better growth and yield compared to the respective

inorganic fertilizer treatment and control. However, there was suppressive effect on germination with

vermicast treatment.

1. Introduction

Ipomoea carnea (family: Convolvulaceae) is

an amphibious toxic weed widely distributed in

India (Konwer et al., 2007; Abbasi and Abbasi,

2010a) and in other tropical and subtropical

countries (Austin and Huaman, 1996). This plant is

native to South America, and it was introduced into

India as an ornamental plant at the end of the 18th

century (Haines, 1925). Since then it has spread

rapidly in the many parts of the country. This weed

colonizes large tracts of water-bodies and land

areas and causing severe damages to the native

vegetation and fauna. In addition to this, the dead

biomass generated from this and other weeds

deposited at benthic zone of water reservoirs, and

undergo anaerobic decomposition which generates

enormous quantity of methane (Abbasi and Abbasi,

2010b). Methane has 25 times the greenhouse

impact of CO2 and so its release to the atmosphere

is of concern (Abbasi and Abbasi, 2010b, 2012).

At present very few effective weed

management tools are available for control (Abbasi

and Abbasi, 2010a). Chemical herbicides enable

control of these weeds quickly and efficiently, but

temporarily. There is also an environmental cost in

using herbicides. Biological control can be an

environmentally safe but selection and maintaining

of host-specific natural enemies makes the process

complex and challenging. The prolific growth and

rapid regeneration of the weed even after cutting,

also defies physical means of destruction and

control (Ganesh et al., 2008). Although, the

102

ipomoea generates huge biomass, it cannot be used

as firewood due to its poor heating value and as it

emits poisonous gases during the burning process

(Konwer et al., 2007). In some part of the country,

this weed is being used for rural housing, and

fencing in agricultural lands. This weed is also used

as laxative, and reported to be traditional healer for

leucoderma and other skin related diseases, and to

provoke menstruation (Jain et al., 2009). However,

only very small quantity of this weed is utilizable

for these purposes and also in very limited

situation. The huge quantity of this weed biomass

can be utilized for generating organic fertilizer such

as vermicompost and compost. This may provide

an answer to the minimization of this weed biomass

accumulation. In addition to this, application of

vermicompost/compost reduces the widespread

deterioration caused to agricultural land due to

rampant use of inorganic fertilizers. Since huge

amount of energy is required for fertilizer

manufacturing and, there is release of greenhouse

gases and other toxic and hazardous wastes at

chemical fertilizer factories, adopting to the organic

farming techniques will be the economically and

environmentally feasible alternative for chemical

fertilizers.

Studies by several authors (Gajalakshmi et

al., 2001, 2002, 2005; Yadav and Garg, 2011;

Makhija et al., 2011) have shown that by

vermicomposting process, most of the weeds can be

converted to vermicast. But it has to be ascertained

whether these vermicast are beneficial to plants.

Such an assessment is particularly necessary for the

weed like ipomoea as it is known to contain

allelopathic compounds (Patel et al., 2009).

Therefore, attempt has been made in this report to

assess the effect of vermicompost generated from

this weed on the germination and growth of cluster

beans (Cyamopsis tetragonoloba). The response of

this plant to vermicompost generated from ipomoea

has been compared with equivalent inorganic

fertilizer treatments.


The experiments were conducted at

Pondicherry University, Puducherry, India

(11°56’N, 79°53’E). This region located on the east

coast of Indian peninsula has a typical maritime

tropical climate with a dissymmetric rainfall. The

mean annual rain fall of this region is about 1300

mm with 57.25 mean rainy days, restricted mainly

during October to December. The experiments were

conducted in 49 liter volume wooden containers (40

cm height with surface area 35 x 35 cm), lined with

high-density polyethylene (HDPE) sheets. The

barren land soil was used in the experiments to

minimize the errors due to the earlier soil practices.

The experimental soil was characterized as sandy

loam soil with low organic carbon and nutrients

content. The leaves of Ipomoea carnea

vermicomposted by an epigeic species, Eudrilus

eugeniae by using direct vermicomposting process

(Makhija et al, 2011), as detailed in chapter 5. The

physico-chemical properties of the vermicast and of

the soil with which it was used are given in Table 1.

The experiments were carried out during July –

October which is ideal for growing cluster beans in

the study area (ICAR, 2011).

For studying the impact of vermicast on

cluster bean, three sets of experiments were

conducted. Based on the nitrogen, phosphorus, and

potassium (NPK) concentrations seen in the

vermicompost (Table 1), and the levels of NPK

needed in the soil for cluster bean (ICAR, 2011), it

was calculated that the cluster bean would require

7.5 t ha-1

of ipomoea vermicompost (VC) for its

optimum growth. In the first set, one batch of

containers was supplied with 7.5 t ha-1

of VC while

two other batches were given 5 and 10 t ha-1

of VC.

Second set comprised of chemical fertilizers (IF)

which were treated with nutrients N, P, K, Ca, Mg,

S, Fe, Mn, Cu, Zn, B, Mo and Cl in concentrations

equivalent to those present in the respective

vermicompost treatments (Table 2). The nutrients

were supplied in two installments: half at the time

103

Table 1. Chemical and physical properties of vermicast

and soil used in the study


of sowing and the rest at the onset of flowering.

The last set had no supplementation of nutrients

and served as control. In this manner 3 sets of

experiments, encompassing seven treatments were

carried out. As each treatment had 36 containers,

each with one plant, 252 containers were utilized

for the main experiment. Another 150 containers

were maintained with all the treatments, as spares

which were used to replace the containers of which

plants were sacrificed for analysis, or in which the

plants happened to have died. The Pusanavabahar

variety cluster bean, which is locally available, was

used. Following the assessment of germination over

eight days, growth and yield were monitored for an

additional three months. Throughout the

experiments, adequate watering was performed to

maintain 20-30% (v/w) moisture. Deweeding was

done periodically. Neem extract was applied as a

mild pesticide in a few instances when pests were

seen infesting the plants.


characteristics

Seed germination was assessed for 8 days

from the day of sowing and was quantified in terms

of germination value (GV) using the formula of

Djavanshir and Pourbeik (1976):

Where GV is germination value; DGS is daily

germination speed which is computed by dividing

cumulative germination percent by the number of

days elapsed since the beginning of the test; GP is

germination percentage at the end of the test, and N

is frequency or number of DGS that are calculated

during the test. The unit for GV is germinated

seeds/day. It is used as an index to statistically

assess the effects of different treatments.

The growth was assessed on the basis of stem

diameter, length of shoot and root, number of

leaves, number of nodules present in the root, the

nodules size and shoot/root biomass for 3 months.

Plants were harvested periodically, and at each

harvest, the fruits were counted and their fresh and

dry weights were determined. To quantify biomass

in terms of dry weight, samples were oven-dried at

85°C to constant weight. Chlorophyll a and b, and

carotenoids were extracted by the method described

by Moran and Porath (1980). One gram of mature

leaf was ground and incubated with N,N-di-methyl

formamide (DMF) for 24 h at 4 °C in dark, shaking

every 6 hours (Moran and Porath, 1980). The

resultant supernatant was read at 470, 647 and 664

nm and the concentration of pigments were


Vermicast Soil

Chemical properties

pH 6.51±0.06 6.30±0.10


356.8±12.1 8.87±0.02


19.8±0.2 2.66±0.02


Phosphorus mg kg -1

37.83±0.60 0.41±0.01

Potassium g kg-1

2.12±0.04 0.40±0.00

Sulphur mg 100g-1

4.64±0.09 0.54±0.01

Calcium g kg-1

15.5±2.6 8.27±0.01

Magnesium g kg-1

4.85±0.09 0.09±0.02

Boron mg kg-1

78.1±2.3 26.9±1.2

Copper mg kg-1

10.6±0.3 5.08±0.15

Iron mg kg-1

270.3±14.7 59.9±2.9

Manganese mg kg-1

117.4±5.0 45.1±2.2

Zinc mg kg-1

107.0±7.7 55.0±2.6

Molybdenum mg kg-1

BDL BDL

Physical properties

Dry weight % 48.4±1.1 94.7±0.1

Bulk density g cm-3

0.26±0.02 1.28±0.00


1.30±0.05 2.70±0.12



mmhos cm-1

6.25±0.07 0.12±0.02

Total porosity % 80.2±1.8 53.1±2.0


104

Table 2. Amount of inorganic fertilizer applied equivalent to vermicast treatment.


applied

Mass %

of nutrient

in applied

compound

10t ha-1

7.5t ha-1

5t ha-1

Concentra

tion of

nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Concentration

of nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Concentration

of nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Nitrogen CH4N2O 46.65 68.22 142.7a 51.16 107.0

a 34.11 71.34

a


Potassium KCl 52.44 10.41 19.85 7.808 14.89 5.206 9.926

Sulphur 26.93 0.223 Nilb 0.167 Nil

b 0.111 Nil

b

Calcium CaCO3 40.04 75.14 187.7 56.36 140.7 37.57 93.83

Magnesium MgO 60.30 23.44 38.88 17.58 29.16 11.72 19.44

Boron Na2B4O7.10H2O 11.34 0.378 3.331 0.283 2.498 0.189 1.665

Copper CuSO4.5H2O 25.45 0.051 0.201 0.038 0.151 0.026 0.101

Iron FeSO4.7H2O 20.09 1.316 6.552 0.987 4.914 0.658 3.276




determined as detailed by Wellburn (1994). The

yield was calculated in the form of harvest index

(HI) which is the ratio of weight of beans per plant

to the above-ground biomass. The yield as reflected

in pod size, pea size, number of peas per pod and

diseased pods, if any, were recorded at each harvest

with randomly selected 50 fruits.

During the course of the experiment, the

plants were infected with bacterial blight

(Xanthomonas campestris) and alternaria blight

(Alternaria spp). Infestation of white fly (Bemisia

tabaci) also occurred. The bacterial blight infection

was seen in the symptoms of black leaf spots,

necrotic lesions at leaf tips, black streaks on

petioles and stems, split stems, and defoliation

(Mihail and Alcorn, 1985; Ren, 2014). Plants were

considered infected by Alternaria spp. when there

were dark brown lesions on leaves with concentric

zonations demarcated with light brown lines. In

severe infections, several spots merged and gave

the leaves a blighted look, eventually causing

defoliation (Orellana and Simmons, 1966;

Yogendra et al., 1995). The number of plants that

died, either with or without any symptoms of

infection, and stunted plants were also recorded.


The analytical methods were the same as

detailed in section 2.3 of chapter 6.



were employed for determining significance of

difference between the results (SPSS version 16;

Softonic, Barcelona, Spain).



The germination value (GV) (F2,32 = 8.223, p

<0.01) and germination percentage (GP) (F2,32 =

33.02, p <0.001) were significantly influenced by

different treatments (Table 3). In all the treatments,

maximum GV was observed on 4th day since the

beginning of the experiment and then there was a

105

decreasing trend. In most of the treatments, lowest

GV was reported on the 8th day. In the VC

treatment, increase in the dose of application has

shown reduction in the germination rate (Table 3).

A maximum of 80% of GP was observed with 5 t

ha-1

VC treatment and lowest GP of 62.9% with 10

t ha-1

treatments. The increasing IF application

equivalent to VC treatment showed increasing trend

in GP. The IF treatment showed maximum GP of

78.6% with 10 t ha-1

treatment followed by 74.3%

and 68.6% with 7.5 t ha-1

and 5 t ha-1

treatments

respectively. However, a maximum GV of 31.9 was

in 7.5 t ha-1

treatment on the 4th day.

This result indicates that, seed germination may be

strongly influenced by the applied IF constituent,

particularly nitrate and ammonium they contains.

As these compounds break dormancy and stimulate

germination (Egley and Duke, 1985; Hilhorst and

Karssen, 2000), increasing the concentration of IF

would have increased the germination success.

Their nature of readily leaching from surface soil

by irrigation water may be the reason for higher

germination at initial days followed by steep

inclination till the end. In the case of VC, a

maximum germination success was recorded in 5 t

ha-1

VC. With increasing dose of VC application a

declining trend of seed germination was observed.

However, in all the VC treatment, GV was much

higher than the control. Few studies have reported

similar suppressive effect on germination with

increasing concentration of VC generated from

kitchen waste, paper waste, yard waste and cattle

dung (Roberts et al., 2007; Warman and AngLopez,

2010; Ievinsh, 2011). Several factors have been

postulated by different authors for the suppressive

effect of VC on germination, such as high

concentrations of gallic acid and chlorogenic acid

(Reigosa et al., 1999), auxin and gibberlellin-like

substances in humic and fulvic acids of

vermicompost (Arancon et al., 2006). In the present

study, presence of some of these components in

excess quantity might be the reason of suppressive

effect on seed germination with increasing dose of

VC.

3.2. Plant growth

The VC and IF application showed a

differential impact on all the plant growth (Table

4). The plants which were treated with VC grew

faster than the IF treated plants and control. The

plants treated with VC showed about 15% higher

stem diameter, 17% longer shoot length, 33%

higher dry weight of shoot, and 16% higher number

of leaves than those of equivalent IF. Even though


by ipomoea vermicast and inorganic fertilizers. Results which do not differ significantly (LSD test; p <0.05)

carry at least one character in the superscript which is common.



Vermicompost

5t ha-1 a

24.70 62.86 17.27 65.71 13.06 68.57 13.06 80.00 10.00 80.00

7.5 t ha-1 ac

22.50 60.00 15.81 62.86 11.99 65.71 10.41 71.43 9.30 77.14

10 t ha-1 b

14.74 48.57 13.06 57.14 10.00 60.00 7.35 60.00 6.17 62.86

Inorganic

fertilizer

equivalent to

the VC

5t ha-1 cb

16.53 51.43 13.06 57.14 11.99 65.71 8.81 65.71 7.35 68.57

7.5 t ha-1 a

31.89 71.43 22.08 74.29 15.33 74.29 11.26 74.29 8.62 74.29

10 t ha-1 a

29.39 68.57 20.41 71.43 15.33 74.29 12.14 77.14 9.65 78.57

Control Nil d 2.50 20.00 2.64 25.71 2.74 31.43 4.26 45.71 3.47 47.14

106

Table 4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth /death in cluster bean plants as impacted by ipomoea


common.

*p<0.05,

**p<0.01,

***p<0.001,


Parameters


Control

F-value

5 t ha-1

7.5 t ha-1

10 t ha-1

5 t ha-1

7.5 t ha-1

10 t ha-1

Type of

fertilizer Amount

Plant growth


b 24.1±2.5

b 16.8±1.4

a 18.4±1.2

a 22.9±2.0

b 11.2±1.7

c 36.70

*** 70.94

***

Shoot length (cm) 144.5±5.0a 197.6±9.8

b 231.6±10.5

c 138.6±5.7

a 162.7±7.3

d 191.4±6.9

b 76.8±5.9

e 70.73

*** 202.3

***

Root length (cm) 55.0±8.0a 41.4±4.1

b 38.3±4.8

b 47.7±5.2

c 47.2±4.3

c 39.6±3.9

b 50.2±8.4

c 2.364

n.s 18.33

***

Number of leaves 98.4±5.5a 120.3±7.7

b 133.8±5.6

c 92.3±3.8

d 108.3±6.0

e 112.2±6.6

e 42.0±5.3

f 162.1

*** 236.9

***


b 129.3±23.2

c 31.9±7.2

d 50.0±11.9

e 37.1±7.3

d 13.5±6.0

f 76.96

*** 15.41

***

Shoot dry weight (g) 51.5±5.6a 89.3±6.6

b 102.6±10.8

c 42.9±4.6

d 59.6±5.2

e 82.8±6.6

f 45.3±5.2

d 18.64

*** 74.71

***

Root dry weight(g) 5.02±0.22a 6.25±0.60

b 11.90±2.14

c 4.60±0.74

a 8.10±1.18

d 6.84±0.93

b 3.21±0.58

f 15.21

*** 40.90

***

Leaf pigments


) 2.17±0.14a 3.21±0.25

b 3.45±0.11

c 1.99±0.09

d 2.69±0.10

e 2.72±0.06

e 1.32±0.05

f 58.35

*** 145.0

***


) 2.21±0.05a 2.30±0.06

b 2.47±0.08

c 2.10±0.14

d 2.28±0.10

ba 2.57±0.08

e 1.45±0.04

f 137.5

*** 370.8

***


) 4.38±0.14a 5.51±0.24

b 5.92±0.16

c 4.09±0.08

d 4.97±0.12

e 5.29±0.10

f 2.77±0.04

g 88.05

*** 329.7

***

Carotenoids (mg g-1

) 0.42±0.04a 0.51±0.05

b 0.55±0.08

b 0.36±0.04

c 0.41±0.05

a 0.53±0.07

b 0.26±0.02

d 38.36

*** 59.70

***

Flowering


b 31.7±11.1

cd 15.7±4.2

e 27.9±8.0

ad 18.1±8.7

ce 0.7±1.0

f 158.0

*** 84.87

***



a 8

a 17

a 9

a 15

a 16

a 1.201

n.s 1.193

n.s

Number of plant died Nila 1

a Nil

a 2

a 2

a 1

a Nil

a 2.873

n.s 0.874

n.s


a 1

a 1

a 5

a 6

a 11

a 2.380

n.s 1.384

n.s

107

the plants from both the VC and IF treatments

received same amount of all the essential nutrients,

there was a drastic difference in its growth, which

probably due to a number of reasons, including

favorable nodules formation, nutrient release

synchronized with plant need, and the contribution

of beneficial plant growth regulators, hormones and

microbes with VC application.

In this experiment, maximum number of

nodules was observed with the plants which were

treated with VC than IF. The increasing dose of VC

application has increased the number of nodules to

the extent of 89%, in comparison with control. In

the case of 5 and 7.5 t ha-1

VC treatments, about

45% higher number of nodules was recorded in

comparison to the equivalent IF treatments and

those of 10 t ha-1

VC showed 89% higher number

of nodules. The results are indicates that VC

application favored nodule formation, in contrast IF

treatments induce depression in nodulation.

Inhibition of nodulation with IF treatments

probably the reason of its high nitrate content

(Carroll and Gresshoff, 1983; Harper and Gibson,

1984; Nie, 1989). van Schreven (1959) had

reported that increasing concentration of urea

application decrease the weight of nodules; even

the lowest concentration of 0.5% exerted this

suppressing effect on clover plants (Trifolium

repens) and lucerne plants (Medicago sativa).

Presence of higher Mn in the IF may also have

contributed to the suppression of nodulation in the

IF treatment (Dobereiner, 1966; Foy et al., 1978).

Even though VC also contains the same inorganic

nutrients, and in the same concentration, as IF, the

release from VC might have been at much slower

rate because of the predominantly organic matrix of

the VC. This might have made nutrients available

to the legume at a rate best suited to promote

nodules formation. The slow release of nutrients by

the VC may also have caused lesser nutrient loss

from the rhizosphere. In addition, supplement of

biologically active substances such as fulvic acids,

humic acids and phytohormones and improvement

in the physical properties of the soil with VC

application also might have contributed to better

plant growth in VC treated plants, which is

discussed in detail in the chapter 6.


Photosynthetic pigments such as chlorophyll and

carotenoid concentration of the plants were

significanly influnced by the applied VC and IF

(p<0.001;Table 4). In general, VC application was

more effective than the respective inorganic

fetilizer treatment. However, in most of the weeks,

VC treatment at the dose of 5 t ha-1

has shown

lesser amount of chlorophyll a and b than the

equivalent inorganic fertilizer treatments. Lower

rate of nutrient application and their poor

availability due to the slower releasing property of

vermicast might have reduced these pigments

concentration with 5 t ha-1

VC treatment. As the

formation of chlorophyll depends on the adequate

supply of nitrogen and iron, the plants which has

supplied with maximum of these nutrients either in

the form of VC and IF had increased its pigment

content. The photoprotection pigments, carotenoids

also found to be synthesized more in the plants

grown in VC treated soil than equivalent IF

treatment.

3.4. Effect on flowering

The VC treated plants produced more number

of flowers in comparison to the equivalent IF

treated plants (p< 0.001) (Table 4). A maximum

number of flowers was recorded with 7.5 t ha-1

treatment with both VC and IF treatments.

Increasing dose fertilizer application showed the

trend of 7.5 t ha-1

> 10 t ha-1

> 5 t ha-1

> control

with VC and equivalent IF treatment. The plants

grown in the soil treated with 7.5 t ha-1

of both VC

and IF showed early flowering than the other

treatments. Adequate availability of nutrient in this

treatment could be the reason for the higher and

early flowering in this study.

108


growth

The plants were infected with bacterial blight

(Xanthomonas campestris), and alternaria blight

(Alternaria spp) and infested with whiteflies

(Bemisia tabaci) maximally during the first two

months (Table 4). The incidence of disease was not

significantly differing with VC and IF application.

However, a noticeable difference was observed

with the amount of nutrient applied in both VC and

IF treatments. The both VC and equivalent IF

supplemented at the dose of 5 t ha-1

showed

maximum number of infected plants.

Moreover, a few plants died during the

experiment, but there was no significant difference

in the number among different treatments (Table 4).

The maximum number of stunted plants was seen

in the first 12 days since beginning of the

experiment. After a month, all the plants grew well.

Inorganic treatment exhibited more number of

stunted growth plants in comparison to the VC

treatment. In IF treatment at the dose of 10 t ha-1

, a

maximum of 6 stunted plants was recorded,

whereas, in all VC treatments lesser number of

plants exhibited stunted growth.

3.6. Effect on yield

The observation on yield attributes is given in

Table 5. The weight and number of harvested pods

from the experimental plants were significantly

different with different form and amount of nutrient

applied (p<0.001) (Table 8). The plants treated with

VC at the dose of 5, 7.5 and 10 t ha-1

produced 6,

20 and 24% higher mass of fruits, respectively in

comparison to its equivalent IF treatments. In both

VC and IF treatments, the weight and number of

pods harvested were maximum at 7.5 t ha-1

;

whereas, increase dose to 10 t ha-1

, reduced the fruit

yield. Reduction of yield in the 10 t ha-1

VC/IF

applications probably due to the excess nutrient

applied in the treatments which is well above the

levels required by a plant may induced higher

immobilization of nutrients in the plant biomass

and less partitioned to fruits (Wada et al., 1989;

Sujatha and Bhat, 2013).

The pods harvested from the VC treated

plants were lengthier, wider and contains more

number of seeds than those of IF treatments. The

seed diameter and thickness were also higher in VC

than the equivalent IF treatments. In both VC and

IF treatments at 7.5 t ha-1

showed these yield

Table 5. Harvest index and yield attributes of plants as impacted by ipomoea vermicast and equivalent inorganic

fertilizers (mean ±SD). Results which do not differ significantly (LSD test; p <0.05) carry at least one character in the

superscript which is common.

*p<0.05, **p<0.01, ***p<0.001,n.s - not significant.

Parameters, average

value

Vermicompost at dose Inorganic fertilizer at dose F-value

5 t ha-1 7.5 t ha-1 10 t ha-1 5 t ha-1 7.5 t ha-1 10 t ha-1 Type of

fertilizer Amount

No.of pod per plant 20.1±8.1a 36.3±10.0b 28.3±7.2c 19.5±6.7a 30.9±7.8c 22.4±7.5a 103.4*** 91.39***

Weight of the pods (g) 110.1±43.8a 201.2±55.9b 159.3±42.4c 103.7±35.7a 161.4±42.7cd 120.8±41.2a 134.8*** 147.8***

Pod length (cm) 14.6±1.8a 15.8±1.6b 15.5±1.6b 14.1±1.6a 14.7±1.9a 14.3±1.7a 22.24*** 6.851**

Pod width (cm) 10.2±1.0a 10.8±0.9bd 16.0±1.3c 10.2±1.3ade 10.4±0.9be 10.8±1.5b 53.32*** 83.730***

Pod thickness (cm) 6.39±0.85a 6.01±0.75b 6.79±0.58c 5.80±0.54b 5.99±1.04b 5.74±0.74b 36.21*** 2.604 n.s

No.of seeds per pod 9.24±0.92a 9.50±1.16a 9.50±1.16a 9.18±1.16a 9.30±1.28a 9.18±1.12a 2.175 n.s 0.728 n.s

Seed diameter (mm) 5.59±1.01a 6.01±0.86b 6.23±1.69c 5.61±0.95a 5.96±0.89b 6.51±0.72d 5.565* 170.9***

Seed thickness (mm) 2.30±0.57a 1.90±0.59b 2.43±0.59c 2.14±0.63d 2.15±0.46d 2.51±0.49e 5.705* 153.3***

Harvest Index % 65.2±40.0abcf 86.9±67.8bdg 49.7±34.3ceh 73.4±51.1ade 87.8±61.4agh 47.0±39.2cf 0.292 n.s 7.713**

109

attributes in better measure than the 5 and 10 t ha-1

treatments, whereas seed diameter and seed

thickness were maximum in 10 t ha-1 treatments.

Even though, VC treated plants given significantly

higher fruit yield, the harvest index (HI) values of

VC treatments are not significantly higher than that

of the IF treated plants. This is because HI is a ratio

of mass of fruits and mass of the above-ground

biomass. In VC treated plants both are higher than

in IF treated plants leading to similar HI. However,

different dose of nutrient application showed

significant influence on HI. Among all the

treatment, the maximum HI was recorded with 7.5 t

ha-1

VC equivalent IF treatment followed by 7.5 t

ha-1

VC treatment; more than 12% higher HI was in

the 7.5 t ha-1

IF treatment.

4. Conclusions

The impact of vermicompost (VC) generated

from the pernicious, allelopathic weed Ipomoea

carnea on germination, growth and fruition of a

botanical species was studied. Response of plants to

the vermicompost was compared with that of an

inorganic fertilizer (IF) which had all the main

macro and micro – nutrients in concentrations

equivalent to the ones present in the VC, and a

control without any nutrient supplement. In this

experiment, nutrient has been supplemented at three

doses, 5, 7.5 or 10 tons per hectare (t ha-1

) and a

legume species, cluster bean (Cyamopsis

tetragonoloba) was used.

In this study, the plant growth in terms of

stem diameter, shoot length, shoot mass, number of

leaves, and leaf pigments were significantly

improved with VC application than with IF. In

addition, VC application enhanced root nodule

formation, and permitted lesser number of stunted

plants; whereas, the IF application suppressed the

root nodulation in the experimental plants. The

plant growth was significantly improved with

increasing dose of both VC and IF application.

However, the yield was maximum in 7.5 t ha-1

VC/IF treatments than the 5 and 10 t ha-1

treatments. Moreover, the root elongation was also

suppressed by excess nutrient application in both

VC and IF treatments. Overall results show that

vermicompost generated from ipomoea had positive

impact on the growth and yield of the plant studied.

In terms of germination, vermicast showed

suppression effect, indicating the presence of some

of the germination inhibitory components in

vermicast.

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112




1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

1.0 1.6 2.1 4.3 5.2 7.1 8.3 12.7 13.7 16.2 16.9 16.9

7.5 t ha-1

1.3 2.1 2.9 3.9 7.6 9.0 11.5 15.3 15.5 22.3 22.5 22.6

10 t ha-1

1.4 2.3 3.2 5.6 8.1 10.1 12.2 17.8 19.2 23.7 24.0 24.1

Inorganic

fertilizer

5 t ha-1

1.2 2.2 2.4 4.5 5.0 6.7 8.6 11.5 13.2 16.6 16.8 16.8

7.5 t ha-1

1.5 2.3 2.7 4.8 5.9 7.9 9.3 14.4 17.4 18.1 18.3 18.4

10 t ha-1

1.5 2.7 2.9 5.7 6.3 8.5 10.2 16.4 20.3 22.7 22.9 22.9

Control Nil 0.7 1.8 2.0 2.3 4.5 5.2 5.8 6.1 1.2 10.8 11.1 11.2



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

4.8 9.5 14.6 23.5 25.2 28.5 37.6 53.1 88.0 130.5 1394.1 144.5

7.5 t ha-1

7.5 11.7 19.7 24.8 32.6 40.5 51.4 59.7 90.2 190.8 192.8 197.6

10 t ha-1

11.5 12.5 21.2 27.4 37.8 44.2 59.8 72.9 115.6 220.1 225.4 231.6

Inorganic

fertilizer

5 t ha-1

6.5 10.5 15.8 17.5 23.7 28.0 34.8 54.0 80.2 122.5 132.8 138.6

7.5 t ha-1

7.5 11.7 16.3 19.1 25.9 29.5 36.9 48.9 94.3 140.5 153.9 162.7

10 t ha-1

10.5 13.5 18.8 21.4 27.2 29.0 41.3 47.2 103.6 175.8 180.2 191.4

Control Nil 3.5 11.4 11.8 12.5 18.0 21.8 23.0 26.0 34.5 42.0 59.3 76.8



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

4.2 16.1 18.5 25.5 27.6 29.2 31.9 32.8 34.6 51.4 54.8 55.0

7.5 t ha-1

5.5 15.4 19.6 21.5 23.6 24.5 25.7 27.1 29.6 37.5 38.5 41.4

10 t ha-1

6.8 21.5 22.5 24.3 25.7 26.2 28.1 29.6 31.6 34.2 36.8 38.3

Inorganic

fertilizer

5 t ha-1

4.9 13.1 19.5 27.3 29.7 33.0 35.9 38.7 41.8 43.1 45.4 47.7

7.5 t ha-1

5.5 13.4 21.4 28.2 30.2 33.2 37.6 40.7 41.5 42.8 45.3 47.2

10 t ha-1

7.6 15.7 18.2 22.3 24.2 25.8 27.1 28.3 32.3 36.8 38.1 39.6

Control Nil 3.8 13.7 14.0 23.1 29.1 29.2 37.4 42.0 45.7 47.5 49.3 50.2

113



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

03 03 04 14 19 33 37 48 55 87 92 98

7.5 t ha-1

03 04 07 19 24 34 42 60 72 105 117 120

10 t ha-1

03 04 07 20 28 43 47 65 81 118 125 134

Inorganic

fertilizer

5 t ha-1

03 03 04 17 21 33 39 58 62 72 85 92

7.5 t ha-1

03 04 07 18 24 33 41 52 68 82 98 108

10 t ha-1

03 04 08 21 27 28 48 59 82 99 107 112

Control Nil 03 03 04 09 15 17 21 25 28 31 35 42



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

Nil 8 11 12 15 20 26 42 38 42 58 61

7.5 t ha-1

Nil 12 13 10 21 24 29 24 42 57 72 90

10 t ha-1

Nil 12 15 15 26 33 35 52 60 59 86 129

Inorganic

fertilizer

5 t ha-1

Nil 3 5 8 9 25 28 39 42 28 29 32

7.5 t ha-1

Nil 12 15 22 27 34 31 30 43 49 42 50

10 t ha-1

6 8 7 4 14 29 18 35 29 31 30 37

Control Nil Nil 0 7 8 12 14 19 11 11 9 16 14



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.031 0.102 0.459 0.981 2.147 4.446 13.067 21.07 25.93 47.58 49.16 51.46

7.5 t ha-1

0.036 0.181 0.367 0.758 2.668 5.755 15.45 24.81 29.18 83.53 88.04 89.31

10 t ha-1

0.054 0.134 0.648 1.065 4.093 7.033 19.31 27.58 37.13 92.62 99.27 102.56

Inorganic

fertilizer

5 t ha-1

0.082 0.120 0.493 0.820 2.105 4.223 12.30 19.88 24.79 40.21 41.72 42.92

7.5 t ha-1

0.047 0.136 0.355 0.647 2.485 5.261 14.94 20.38 27.34 56.24 57.93 59.64

10 t ha-1

0.008 0.094 0.539 0.602 3.827 6.875 17.11 24.11 34.60 74.55 79.62 82.81

Control Nil 0.032 0.109 0.205 0.395 0.955 1.490 4.572 6.799 12.11 41.50 44.08 45.32

114





1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.398 0.779 1.095 1.503 1.716 1.791 1.645 1.798 1.823 1.925 2.092 2.167

7.5 t ha-1

0.414 1.642 1.955 2.115 2.326 2.401 2.578 2.774 2.823 2.890 3.172 3.211

10 t ha-1

0.543 1.795 2.093 2.375 2.583 2.741 2.874 3.108 3.195 3.284 3.376 3.452

Inorganic

fertilizer

5 t ha-1

0.378 0.914 1.218 1.649 1.789 1.589 1.674 1.699 1.718 1.825 1.959 1.990

7.5 t ha-1

0.307 1.458 1.541 1.632 1.735 1.77 1.823 2.236 2.419 2.497 2.556 2.691

10 t ha-1

0.541 1.618 1.481 1.781 1.974 2.122 2.249 2.364 2.592 2.614 2.628 2.717

Control Nil 0.103 0.465 0.64 0.907 1.128 1.164 1.177 1.193 1.201 1.224 1.238 1.316

Table 4I. Leaf chlorophyll b (mg g-1



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.102 0.627 0.948 1.06 1.18 1.254 1.635 1.806 1.902 2.132 2.160 2.207

7.5 t ha-1

0.148 0.757 0.978 1.209 1.497 1.712 1.964 2.024 2.100 2.195 2.222 2.296

10 t ha-1

0.291 0.999 1.167 1.249 1.591 1.854 2.072 2.168 2.131 2.205 2.234 2.474

Inorganic

fertilizer

5 t ha-1

0.179 0.304 0.963 1.118 1.244 1.418 1.64 1.823 1.825 2.014 2.060 2.097

7.5 t ha-1

0.141 0.475 1.021 1.393 1.571 1.846 1.91 2.116 2.176 2.182 2.220 2.278

10 t ha-1

0.374 0.52 1.134 1.586 1.618 1.987 2.128 2.351 2.404 2.481 2.517 2.568

Control Nil 0.032 0.155 0.223 0.504 1.612 1.935 1.065 1.409 1.510 1.532 1.544 1.451


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.005 0.024 0.039 0.066 0.125 0.607 0.984 1.642 3.094 4.751 4.882 5.024

7.5 t ha-1

0.001 0.026 0.042 0.086 0.276 0.847 1.446 2.044 3.916 5.983 6.034 6.248

10 t ha-1

0.005 0.019 0.057 0.084 0.283 0.621 1.349 2.323 6.800 9.019 9.275 11.937

Inorganic

fertilizer

5 t ha-1

0.000 0.022 0.040 0.072 0.174 0.501 0.728 1.847 2.171 4.192 4.301 4.605

7.5 t ha-1

0.003 0.024 0.042 0.089 0.226 0.738 1.203 2.403 6.064 7.652 7.824 8.102

10 t ha-1

0.000 0.012 0.029 0.064 0.198 0.515 1.004 1.077 5.903 6.061 6.489 6.841

Control Nil 0.001 0.014 0.016 0.029 0.102 0.256 0.289 0.981 1.763 2.929 3.160 3.214

115




1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.500 1.406 2.043 2.563 2.896 3.045 3.280 3.604 3.725 4.057 4.252 4.374

7.5 t ha-1

0.562 2.399 2.933 3.324 3.823 4.113 4.542 4.798 4.923 5.085 5.394 5.507

10 t ha-1

0.834 2.794 3.26 3.624 4.174 4.595 4.946 5.276 5.326 5.489 5.610 5.926

Inorganic

fertilizer

5 t ha-1

0.557 1.218 2.181 2.767 3.033 3.007 3.314 3.522 3.543 3.839 4.019 4.087

7.5 t ha-1

0.448 1.933 2.562 3.025 3.306 3.616 3.733 4.352 4.595 4.679 4.776 4.969

10 t ha-1

0.915 2.138 2.615 3.367 3.592 4.109 4.377 4.715 4.996 5.095 5.145 5.285

Control Nil 0.135 0.62 0.863 1.411 2.740 3.099 2.242 2.602 2.711 2.756 2.782 2.767




1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.101 0.139 0.197 0.261 0.292 0.314 0.335 0.364 0.371 0.398 0.405 0.424

7.5 t ha-1

0.126 0.235 0.289 0.322 0.357 0.398 0.423 0.456 0.477 0.483 0.491 0.512

10 t ha-1

0.166 0.244 0.302 0.384 0.416 0.427 0.438 0.479 0.489 0.503 0.528 0.547

Inorganic

fertilizer

5 t ha-1

0.16 0.192 0.231 0.249 0.276 0.288 0.291 0.301 0.312 0.333 0.348 0.361

7.5 t ha-1

0.104 0.151 0.247 0.275 0.291 0.317 0.341 0.366 0.371 0.382 0.400 0.412

10 t ha-1

0.107 0.266 0.382 0.416 0.427 0.435 0.442 0.456 0.482 0.499 0.529 0.530

Control Nil 0.102 0.122 0.136 0.158 0.164 0.191 0.203 0.218 0.224 0.236 0.248 0.261

COMPARATIVE EFFICACY OF VERMICOMPOSTED

PAPER WASTE AND INORGANIC FERTILIZER ON

SEED GERMINATION, PLANT GROWTH AND

FRUITION OF CLUSTER BEAN (CYAMOPSIS

TETRAGONOLOBA)

Chapter

8

117


Journal of Applied Horticulture, 16 (1), 40 – 45, 2014.

CChhaapptteerr 88

Comparative efficacy of vermicomposted paper waste and

inorganic fertilizer on seed germination, plant growth and

fruition of cluster bean (Cyamopsis tetragonoloba)

Abstract

The aim of the present study was to assess the influence of vermicompost generated from the paper waste

spiked with cow dung slurry on the germination, plant growth and fruition of cluster bean (Cyamopsis

tetragonoloba). Two kinds of treatments were studied: (i) vermicast was applied to the soil at the rates of 5,

7.5, 10 t ha-1

, (ii) amounts of essential nutrients equivalent to those present in the vermicast treatments in

inorganic form was amended to the soil. There was a control with only soil without any nutrient supplement.

The finding is in contrast to the reports on the beneficial impacts of vermicast on plant growth. In the present

study the inorganic fertilizer treatment exhibited better seed germination and plant growth than the

equivalent vermicast treatments. The results indicate that the dose of vermicompost used in the present study

was not sufficient to satisfy the nutrient demand of plant species studied. Additional fertilization would have

improved the crop productivity.

1. Introduction

Paper waste generation has been continually

increasing over the past years due to increasing

population, industrialization, urbanization and

literacy. In India, the paper consumption is about

8.5 kg per capita per year and it is 0.81-5.8 % of

municipal solid waste (MSW) (Gupta and Garg,

2009; www.indiastat.com). Due to the absence of

waste segregating practices, the paper waste is

dumped along with all other kinds of waste in open

and poorly managed landfills, which is very

common practice in most of the cities in India. The

improper disposal of this degradable waste may

lead to long term threat to the environment and

public health, such as the risk of ground water

pollution due to leachate seepage, fugitive

greenhouse gas emission contributing to climate

change and odor pollution caused by non-methane

organic compounds, which is direct harassment to

adjacent communities (Zhang et al., 2012). Also,

open dumping of wastes facilitates the breeding for

disease vectors such as flies, mosquitoes,

cockroaches, rats, and other pests (CPCB, 2000).

Paper and cardboard have a relatively high

heating value, similar to wood, and this energy

utilized via incineration can be transformed into

electricity (Villanueva and Wenzel, 2007).

However, incineration is not very much practiced in

India due to lack of awareness and absence of waste

segregating practices. The paper waste when mixed

with other moist organic waste and inert material

reduces its calorific value (Negi and Suthar, 2013).

In recent years, due to shortage of raw material,

waste paper is preferred for paper production. Also,

recycling of paper consumes only 40% of the

energy in comparison to the process based on other

raw materials (Gupta et al., 1998). However, the

paper recycling industries prefer to use imported

waste paper because of its better quality in terms of

fibre strength and also due to inadequate domestic

118

supply owing to the unorganized collection of

waste paper within the country. In addition, the

yield from imported waste paper can be as high as

90%, whereas the available waste paper in India

gives yield less than 50% due to shorter fibre length

(IPMA, 1996). Nevertheless, if waste papers are

segregated at the source itself, it could be the input

material for paper recycling units. However, due to

lack of efficient waste management service, the

paper waste invariably finds its way to the MSW at

the end.

The huge generation of these paper wastes

can be treated by vermicomposting which convert

the waste into useful end product that can be used

as a soil amendment (Sinha et al., 2010). Unlike

recycling and incineration, the biological

composting process not affected either by quality of

waste paper or when mixed with other organic

wastes. Processing of paper waste through

vermicomposting may provide an answer to the

minimization of waste accumulation and also to

widespread deteriorated agricultural land due to

rampant use of inorganic fertilizers. It is well

established that vermicompost application have

beneficial impact on soil physical, chemical and

biological properties and can increase the

germination, plant growth and yield in both natural

and agricultural ecosystem (Edwards and Bohlen,

1996). These beneficial effects have been attributed

to improvement in soil properties and structure, to

greater availability of mineral nutrients to plants

(Edwards, 1998). In addition to this, vermicompost

contain plant growth regulating components,

including plant growth hormones and humic acids

that are reported to be responsible for promoting

germination, growth and yields of plants, in

response to vermicompost applications or

substitutions, independent of the nutrients they

contain (Tomati et al., 1988; Muscolo et al., 1999;

Atiyeh et al., 2002; Arancon et al., 2003, 2006).

However, the vermicompost generated from

waste paper causes apprehension towards the

beneficial impact on plant growth due to the low

nutrient content of this substrate. Therefore, attempt

has been taken to investigate the beneficial impact

of vermicompost generated from the paper waste

(VC) on the germination, plant growth and yield of

cluster bean, a vegetable crop. This plant has

chosen due to their drought tolerance which reduces

the error due to the other environmental factors.

Moreover as it is a leguminous plant, influence of

vermicast and inorganic fertilizers on nodules

formation and its growth can be revealed. In

addition, to evaluate the possible non-nutrient (i.e.

hormones and other growth regulating components)

dependent effect of vermicast over inorganic

fertilizers, all essential nutrients present in the

vermicast were supplied in inorganic form (IF) to

the plants, and the response of plants to the

different fertilizers is briefed in this paper.


2.1. Study area

This experiment was conducted at

Pondicherry University, Puducherry, India,

located on the east coast of Indian peninsula

(11°56’N, 79°53’E). The climate of the

experimental site is typical maritime of tropical

climate with a disymmetric rainfall. The

average annual rain fall is about 1300 mm with

57.25 mean rainy days, and around 60% of the

total rainfall is received during period of

October to December through the north–east

monsoon.

2.2. Treatments

The experiment was set up in 49 liter volume

wooden containers (40 cm height with surface area

35 x 35 cm), lined with high-density polyethylene

(HDPE) sheets. These containers were filled with

low fertile barren land soil collected inside the

Pondicherry University campus to reduce the errors

due to previous soil practices. The experimental

soil was characterized as sandy loam soil and its

physico-chemical properties are shown in Table 1.

119

Table 1. Chemical and physical properties of

vermicast and soil used in the study.


The experiment was conducted during the Kharif

season which is best time for sowing the cluster

beans in south India. The Pusanavabahar variety

was used which is locally available in the

experimental area. The vermicompost was

generated from the paper waste spiked with cow

dung slurry by employing an epigeic species,

Eudrilus eugeniae Kinberg. The VC was applied to

the plant growth containers at the rate of 5, 7.5 and

10 t ha-1

. In another set, an equivalent amount of all

major and minor nutrients present in vermicompost

was supplied as inorganic chemical form to check

the efficiency of vermicast over the inorganic

fertilizer.

In the IF treatment, the primary nutrients N, P

and K, secondary nutrients Ca, Mg and S, and

micronutrients of Fe, Mn, Cu, Zn, B, Mo and Cl

were applied to an equivalent amount of 5, 7.5 and

10 t ha-1

VC treatment. The chemical fertilizers

were applied in the form of urea, di-ammonium

phosphate, potash, CaCO3, MgO, Na2B4O7, CuSO4,

FeSO4, MnSO4 and ZnCl2. Besides these

treatments, one more set was maintained without

any supplementation, as control i.e only soil. The

nutrients were supplied in two phases. The first

phase was at the time of sowing which comprised

half of the total nutrients. The second

supplementation of nutrients was done at the time

of flowering of plants.


characteristics

Two seeds per container, totally 72 seeds per

treatment were sown in all the containers. Seeds

were considered germinated when they exhibited

radical extension of >3 mm. Counts of germinated

seeds were made daily up to eight days to

determine the germination rate in terms of

germination percentage (GP) and germination value

(GV) by method described by Djavanshir and

Pourbeik (1976). The GV is used as a comparative

index to statistically assess the effects of the

treatments. After germination, one plant for each

compartment was maintained by removing excess

plant grown from the sown seeds. A separate

nursery was also maintained with all VC and IF

treatments. Healthy plants from the nursery were

transplanted to containers where seeds failed to

germinate. Adequate amount of water was provided

during the experiment. Deweeding was done

manually. In few instances when pests were seen,

organic pesticides such as neem extract and cow

urine were used.

The plant height, length of shoot and root,


Vermicast Soil

Chemical properties

pH 7.83±0.05 6.30±0.10


259.6±8.2 8.87±0.02


11.7±0.4 2.66±0.02


Phosphorus mg kg -1

71.40±3.31 0.41±0.01

Potassium g kg-1

1.98±0.10 0.40±0.00

Sulphur mg 100g-1

0.40±0.04 0.54±0.01

Calcium g kg-1

15.9±1.4 8.27±0.01

Magnesium g kg-1

7.13±0.07 0.09±0.02

Boron mg kg-1

7.28±0.09 26.9±1.2

Copper mg kg-1

16.9±1.0 5.08±0.15

Iron mg kg-1

208.3±11.4 59.9±2.9

Manganese mg kg-1

63.8±1.8 45.1±2.2

Zinc mg kg-1

94.1±2.2 55.0±2.6

Molybdenum mg kg-1

BDL BDL

Physical properties

Dry weight % 49.6±1.9 94.7±0.1

Bulk density g cm-3

0.24±0.01 1.28±0.00


1.21±0.03 2.70±0.12



mmhos cm-1

2.96±0.04 0.12±0.02

Total porosity % 80.08±1.0 53.1±2.0


120

Table 2. Amount of inorganic fertilizer applied equivalent to vermicast treatment.


applied

Mass %

of nutrient

in applied

compound

10t ha-1

7.5t ha-1

5t ha-1

Concentra

tion of

nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Concentration

of nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Concentration

of nutrient in

VC

(kg ha-1

)

Amount

of IF

applied

(kg ha-1

)

Nitrogen CH4N2O 46.65 30.39 65.15a 22.79 48.86

a 15.19 32.57

a


Potassium KCl 52.44 9.806 18.70 7.354 14.02 4.903 9.348

Sulphur 26.93 0.020 Nilb 0.015 Nil

b 0.010 Nil

b

Calcium CaCO3 40.04 78.98 197.2 59.23 147.9 39.49 98.62

Magnesium MgO 60.3 35.36 58.63 26.52 43.97 17.68 29.31

Boron Na2B4O7.10H2O 11.34 0.036 0.318 0.027 0.239 0.018 0.159

Copper CuSO4.5H2O 25.45 0.084 0.329 0.063 0.247 0.042 0.165

Iron FeSO4.7H2O 20.09 1.033 5.141 0.774 3.855 0.516 2.570




number of leaves, stem diameter, number of

nodules present in the root and their size, biomass

of shoot and root and its dry weight were recorded

with randomly collected samples at each week.

Leaf chlorophyll a, b and carotenoids content were

estimated photometrically by using N,N-di-Methyl

formamide (DMF) as extractent (Moran and Porath,

1980; Wellburn, 1994). Throughout the study, the

disease incidence, and number of flowers produced

were recorded.







difference between the results. The statistical

analysis was carried out using SPSS software

(version 16). The level of significance was at p <

0.05.



In both VC and IF treatments, maximum GV

was observed on fourth day since the experiment

began and then a decreasing trend was observed as

the days progressed. The results of GV did not

show any significant difference with different

forms nor amount of nutrients applied. However,

both forms of fertilizer and amount of nutrient

applied showed significantly higher GV than the

control (p<0.001), and it was more than 10 folds

higher in most of the cases (Table 4). In the VC

treatment maximum GP of 88.6% was observed

with both 7.5 and 10 t ha-1

treatment and maximum

GV of 40 was also recorded in these treatments.

The VC treatment at dose of 5 t ha-1

showed lowest

GP of 74.3% among both VC and IF treatments. In

the IF treatment, an increasing trend in GP was

observed with increasing dosage of fertilizer

application. In this treatment, GP of 88.6, 91.4 and

94.3 % recorded with 5, 7.5 and 10 t ha-1

treatment

on the eighth day and, a maximum GV of 42.9 was

in 7.5 t ha-1

treatments. With all the dose of nutrient

121

application, the VC had shown lesser germination

rate than their respective inorganic fertilizer

treatments (Table 3).

Other studies on influence of VC on seed

germination either showed inhibitory or stimulation

effect with low concentration of VC substitution in

growth media. The discrepancy in the response to

vermicompost depends on plant species which

reacts differently to the concentration of

vermicompost application (Atiyeh et al., 2000;

Edwards et al., 2004). The different organic

substrate used for vermicomposting also changes

the quality of vermicast (Zaller, 2007). Ievinsh

(2011) reported inhibitory effect similar to the

present study on beetroot, beans and pea at low

dose of manure-based vermicompost (5-10%), but

germination dramatically increased with the

increase of vermicompost concentration. The

higher germination in IF treatments may be due to

nitrate which would have been converted from the

urea applied as nitrogen source. These constituents

are breakers of dormancy and also stimulator of

germination (Egley and Duke, 1985; Hihorst and

Karssen, 2000). Since the nitrate readily leaches

from surface soil by irrigation water, the

concentration of nitrate would have declined as the

days progressed, and this may be the reason for

higher germination at initial days followed by steep

inclination till the end.

3.2. Plant growth

There was a differential impact of the vermicast

and inorganic fertilizer treatment on all the

morphological parameters studied (Table 3). The

plants which were treated with IF grew faster than

the VC treated plants. The IF treatment equivalent

to 10 t ha-1

VC recorded higher stem diameter,

length, fresh and dry weight of shoot, number of

leaves, number and size of nodules than VC

treatment and control. The highest of these

parameters: 14.4 mm stem diameter, shoot length of

134 cm, 72 numbers of leaves, 182.1 g of shoot

fresh weight and 52.4 g shoot dry weight was

recorded in the plants treated with IF treatment

equivalent to 10 t ha-1

paper waste VC. Plants

treated with VC at the rate of 10 t ha-1

showed a

maximum of 14.1 mm of stem diameter; 121 cm

shoot length, 65 numbers of leaves, 180 g of shoot

fresh weight and 57.3 g of shoot biomass.

Many authors have reported that high dose of

inorganic fertilizer treatments induce depression in

nodules formation in soybean, chick pea and lupins

(van Schreven, 1959; Carroll and Gresshoff, 1983;


by paper waste vermicast and inorganic fertilizers. Results which do not differ significantly (LSD test; p

<0.05) carry at least one character in the superscript which is common.



Vermicompost

5t ha-1 a

24.70 62.86 17.27 65.71 15.33 74.29 11.26 74.29 8.62 74.29

7.5 t ha-1 b

40.00 80.00 27.46 82.86 20.41 85.71 16.01 88.57 12.26 88.57

10 t ha-1b

40.00 80.00 29.38 85.71 21.79 88.57 16.01 88.57 12.26 88.57

Inorganic

fertilizer

equivalent to

the VC

5t ha-1 b

40.00 80.00 27.46 82.86 20.41 85.71 16.01 88.57 12.26 88.57

7.5 t ha-1 b

42.91 82.86 29.38 85.71 20.41 85.71 14.99 85.71 13.06 91.43

10 t ha-1 b

40.00 80.00 33.44 91.43 24.70 94.29 18.14 94.29 13.89 94.29

Control Nil c 2.50 20.00 2.64 25.71 2.74 31.43 4.26 45.71 3.47 47.14

122

Table 4. Plant growth, leaf pigments, flowering, disease incidence, and retarded growth/death in cluster bean plants as impacted by paper waste


common.

*p<0.05,

**p<0.01,

***p<0.001,


Parameters


Control

F-value

5 t ha-1

7.5 t ha-1

10 t ha-1

5 t ha-1

7.5 t ha-1

10 t ha-1

Type of

fertilizer Amount

Plant growth


bc 14.1±1.2

de 11.9±0.9

ab 13.1±1.4

cd 14.4±2.0

e 11.2±1.7

a 5.634

** 22.55

***

Shoot length (cm) 96.8±4.1a 118.0±10.6

b 121.0±7.9

b 108.4±7.3

c 119.1±4.7

b 134.3±10.9

d 76.8±5.9

e 54.58

*** 97.36

***

Root length (cm) 59.2±9.2a 72.1±14.9

b 83.4±6.1

c 55.7±11.0

ad 89.1±15.5

c 50.1±7.8

d 50.2±8.4

d 7.178

** 14.88

***

Number of leaves 58.8±4.2a 62.2±3.2

ab 64.8±5.7

bc 51.2±6.0

d 68.3±5.4

ce 71.8±4.0

e 42.0±5.3

f 36.49

*** 66.51

***


b 45.3±8.4

b 38.4±11.8

b 38.3±6.6

b 37.1±14.0

b 13.5±6.0

c 23.45

*** 19.26

***

Shoot dry weight (g) 37.1±3.8ab

47.6±5.7c 57.3±6.5

d 39.8±4.0

b 43.2±3.8

be 52.4±6.2

f 45.3±5.2

ce 0.699

n.s 36.92

***

Root dry weight(g) 4.78±0.29a 6.82±0.20

b 7.22±0.55

bc 7.53±0.62

c 9.39±0.86

d 12.7±1.7

e 3.21±0.58

f 74.94

*** 35.04

***

Leaf pigments


) 1.83±0.12a 1.92±0.15

a 1.87±0.10

a 1.84±0.08

a 2.24±0.14

b 2.29±0.12

b 1.32±0.05

c 98.03

*** 57.51

***


) 1.61±0.10ab

1.63±0.11bc

2.05±0.17b 1.51±0.15

ad 2.12±0.19

c 2.28±0.11

e 1.45±0.04

d 14.77

*** 52.62

***


) 3.44±0.12ab

3.55±0.22a 3.92±0.22

c 3.35±0.17

b 4.36±0.23

d 4.57±0.17

e 2.77±0.04

f 45.88

*** 63.60

***

Carotenoids (mg g-1

) 0.29±0.04a 0.46±0.06

b 0.46±0.08

b 0.36±0.10

c 0.36±0.07

c 0.37±0.05

c 0.26±0.02

a 13.17

*** 15.32

***

Flowering


b 3.61±1.99

bc 3.17±1.70

b 4.08±2.47

c 4.39±2.56

c 0.72±1.02

a 34.90

*** 27.57

***



a 21

a 8

a 14

a 12

a 16

a 2.221

n.s 0.378

n.s

Number of plant died Nila 1

a Nil

a 1

a Nil

a Nil

a Nil

a 0.162

n.s 0.490

n.s


a 1

a 1

a 6

a 11

a 11

a 1.276

n.s 0.749

n.s

123

Harper and Gibson, 1984; Nie, 1989). The finding

of the present study contradicts with the previous

reports on suppression of nodules formation with IF

treatment. There was no significant difference in

the number and size of nodules between VC and IF

treatments. The results indicates that there was no

suppression or stimulation of nodules formation

with both VC and IF treatment. The reason may

that very low amount of nutrient has been supplied

to the plants in both VC and IF treatments. In

addition to this, slow releasing property of

vermicast might have reduced the nutrient

availability to the plants. In the VC treated

containers, it was observed that the applied VC did

not disintegrate till the end of the experiment

period. This stable nature of VC might have slow

down the nutrient release to plants. It might be the

reason for lower plant growth in the VC treatment

than the equivalent IF treatment.


The photosynthetic pigments such as

chlorophyll and carotenoid in the leaves of plants

amended with vermicast and inorganic fertilizer

highly correlated with amount of nutrient applied

(Table 3). In general, IF application was more

effective than respective VC treatment. Lower rate

of nutrient application and their poor availability

due to the slower releasing property of vermicast

may be the reason for lower photosynthetic

pigments in this treatment compared to IF treated

plants. In the VC treatment, the maximum total

chlorophyll content of 3.92 mg g-1

was recorded

with the dose of 10 t ha-1

followed by 3.55 and 3.44

mg g-1

with 7.5 and 5 t ha-1

treatments respectively.

In the case of IF treatment, the maximum total

chlorophyll content of 4.57 mg g-1

was recorded in

10 t ha-1

treatment followed by 4.35 and 3.35 mg g-1

with 7.5 and 5 t ha-1

IF treatment. The control

treatment showed lowest chlorophyll content of

2.77 mg g-1

at the end of the experiment.

Carotenoids pigment exhibited similar trend of

results. The increasing nutrient availability with

increasing dose of fertilizer application can be

attributed to the formation of leaf pigments

(Mengel and Kirkby, 1987; Shadchina and

Dmitrieva, 1995; Ruza, 1996; Tejada et al., 2007).


growth

In the first two months of the experimental

period many plants were infected with bacterial

blight (Xanthomonas campestris), and alternaria

blight (Alternaria spp) and infested with whiteflies

(Bemisia tabaci). As a consequence some plants

died and some did not exhibit normal growth

(Table 3). The observed disease incidence and plant

mortality was not significantly differing between

different forms of nutrient application (vermicast

and inorganic fertilizer). The number of stunted

plants was maximum in the beginning of the

experiment. After a month, all the plants grew well.

Inorganic treatment showed more stunted plants in

comparison to the VC treatment. In IF treatment

totally 18 stunted plants were recorded, whereas in

the case of VC treatment at different doses, two or

less stunted plants were recorded.

3.5. Effect on flowering

The IF treated plants were produced more

number of flowers in comparison to the equivalent

VC treated plants (p<0.001; Table 5). A maximum

was recorded with 10 t ha-1

treatment with both VC

and IF treatments. Increasing dose of both fertillizer

application showed the trend with maximum

number of flowers: 10 t ha-1

> 7.5 t ha-1

> 5 t ha-1

>

control. The plants grown in the soil treated with

10 t ha-1

of both VC and IF showed early flowering

than the other treatments. Increased in availability

of nutrient in this treatment could be the reason for

the higher and early flowering in this treatment.

3.6. Effect on yield

In the VC, IF treated plants and control no

vegetables were produced. The reason may be due

to inadequate availability of minor nutrients to the

plants in all the treatments. In this study, low fertile

124

soil collected from barren land was used to

minimize the errors due to the previous soil

practices in the experimental results. The soil was

characterized as very low nutrient soil. Therefore,

the growth and yield of plants relied on the applied

nutrient either in the form of VC or IF. The minor

nutrient applied would have been exhausted thereby

impeding the fruiting in all the treatments.

4. Conclusions

The results obtained from this experiment

shows that application of VC generated from the

paper waste had no beneficial impact on growth of

cluster bean plant. Moreover, the IF treatment

exhibited better seed germination and plant growth

than the equivalent VC treatment. This indicates

that the slow releasing property of VC might have

led to depletion of nutrient availability to the plants.

In this experiment, increasing dose of both

vermicast and inorganic fertilizer improved the

germination rate and plant growth parameters.

However, the application of inorganic fertilizer did

not suppress the root nodules, which is

contradictory to the previous reports (van Schreven,

1959; Nie, 1989). There was no significant

difference in the number and size of nodules with

different treatments. In addition to this, the plants

did not fructify in all the treatments. The low fertile

experimental soil and insufficient nutrient

application might have impeded the production of

vegetables in the present study.

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127




1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

1.2 1.6 1.9 2.5 3.9 5.1 5.6 6.3 9.7 10.2 10.9 10.9

7.5 t ha-1

1.6 1.7 1.8 3.8 4.1 5.6 5.8 6.7 11.2 11.9 12.3 12.8

10 t ha-1

1.7 1.8 2.2 3.3 3.8 3.9 5.3 8.6 11.9 12.6 13.3 14.1

Inorganic

fertilizer

5 t ha-1

1.7 1.8 2.1 2.9 3.2 4.4 5.3 6.1 10.1 11.1 11.4 11.9

7.5 t ha-1

1.6 1.6 1.8 3.1 4 4.1 5.8 6.1 10.4 11.5 11.9 13.1

10 t ha-1

1.2 1.5 1.2 3.7 5.5 8.1 8.6 9.1 11.4 13.1 13.6 14.4

Control Nil 0.7 1.8 2.0 2.3 4.5 5.2 5.8 6.1 1.2 10.8 11.1 11.2



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

14.2 22.3 23.1 25.4 26.5 33.7 41.3 55.7 63.5 73.6 89.1 96.8

7.5 t ha-1

14.1 21.5 21.5 23.4 26.1 39.0 62.4 65.2 77.5 82.3 103.6 118

10 t ha-1

13.1 21.0 14.6 18.6 27.5 33.0 52.5 72.1 98.7 105 113 121

Inorganic

fertilizer

5 t ha-1

11.5 17.1 18.5 22.0 24.6 33.4 34.9 38.0 56.0 67.0 95 108

7.5 t ha-1

14.4 18.5 20.8 25.9 32.0 38.0 48.3 56.2 82.0 96.0 113 119

10 t ha-1

7.2 10.4 14.5 17.6 25.9 37.4 41.2 93.6 117 121 129 134

Control Nil 3.5 11.4 11.8 12.5 18.0 21.8 23.0 26.0 34.5 42.0 59.3 76.8



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

3.4 17.4 22 24.2 34.5 38.1 41.2 42 48.6 51.2 55.3 59.2

7.5 t ha-1

9.3 22 24.8 25.2 28.6 35.5 53 56.4 62.7 65.3 64.7 72.1

10 t ha-1

11 12.9 19.4 21.2 29.1 29.8 52.5 61.8 63.7 68.9 80.1 83.4

Inorganic

fertilizer

5 t ha-1

8.2 11.9 18 19.2 20.2 22.7 24.2 29 36.1 47.4 44.8 55.7

7.5 t ha-1

7.4 13.5 19.6 20.4 31.2 38.1 52.5 69.2 79.6 79 85.3 89.1

10 t ha-1

9.6 12.1 14.2 23.6 25 28.4 43.2 43.2 55 57.2 65.8 50.1

Control Nil 3.8 13.7 14.0 23.1 29.1 29.2 37.4 42.0 45.7 47.5 49.3 50.2

128



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

03 04 04 09 09 18 23 27 37 42 53 59

7.5 t ha-1

03 04 04 12 16 20 31 33 47 51 58 62

10 t ha-1

03 04 05 06 15 23 31 37 46 59 61 65

Inorganic

fertilizer

5 t ha-1

03 04 04 07 09 19 22 27 36 38 49 51

7.5 t ha-1

03 04 04 09 20 22 28 35 54 59 63 68

10 t ha-1

03 03 03 06 18 33 37 48 59 66 69 72

Control Nil 03 03 04 09 15 17 21 25 28 31 35 42



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

Nil 5 7 14 15 18 20 21 18 21 27 28

7.5 t ha-1

Nil 7 11 14 19 25 28 31 28 34 41 39

10 t ha-1

Nil 2 7 9 12 19 30 35 33 39 37 45

Inorganic

fertilizer

5 t ha-1

Nil 3 7 9 12 21 21 33 24 27 26 38

7.5 t ha-1

Nil 3 9 12 15 20 20 22 31 29 38 37

10 t ha-1

Nil 4 8 9 15 17 19 29 28 22 37 34

Control Nil Nil 0 7 8 12 14 19 11 11 9 16 14



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.05 0.11 0.15 0.29 0.38 0.74 1.29 2.35 14.2 22.6 36.7 37.1

7.5 t ha-1

0.05 0.16 0.23 0.70 0.77 0.98 4.72 5.57 15.3 34.1 46.2 47.6

10 t ha-1

0.07 0.15 0.38 0.66 0.78 1.49 3.87 11.7 17.6 43.0 51.8 57.3

Inorganic

fertilizer

5 t ha-1

0.05 0.09 0.16 0.27 0.36 1.08 1.58 2.07 18.9 23.5 33.7 39.8

7.5 t ha-1

0.08 0.13 0.17 0.44 1.14 1.22 3.11 3.70 19.4 30.1 35.8 43.2

10 t ha-1

0.05 0.09 0.18 0.41 1.37 3.23 6.06 9.71 20.6 33.9 45.3 52.4

Control Nil 0.03 0.11 0.21 0.40 1.0 1.49 4.57 6.80 12.1 41.5 44.1 45.3

129





1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.244 0.436 1.064 1.222 1.564 1.659 1.662 1.701 1.726 1.735 1.798 1.825

7.5 t ha-1

0.225 0.516 0.980 1.201 1.377 1.654 1.678 1.759 1.769 1.802 1.858 1.916

10 t ha-1

0.336 0.758 1.154 1.470 1.570 1.727 1.763 0.769 1.813 1.841 1.871 1.866

Inorganic

fertilizer

5 t ha-1

0.388 0.544 0.992 1.184 1.434 1.663 1.766 1.783 1.819 1.826 1.837 1.840

7.5 t ha-1

0.368 0.755 1.083 1.169 1.477 1.770 1.962 2.059 2.096 2.115 2.148 2.236

10 t ha-1

0.568 0.847 1.044 1.172 1.678 2.080 2.179 2.179 2.197 2.206 2.225 2.291

Control Nil 0.103 0.465 0.64 0.907 1.128 1.164 1.177 1.193 1.201 1.224 1.238 1.316

Table 4I. Leaf chlorophyll b (mg g-1



1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.113 0.311 0.623 0.971 1.097 1.375 1.390 1.421 1.475 1.519 1.590 1.613

7.5 t ha-1

0.242 0.310 0.827 0.752 1.032 1.293 1.369 1.401 1.460 1.593 1.616 1.629

10 t ha-1

0.210 0.517 0.970 1.111 1.152 1.440 1.463 1.790 1.875 1.826 1.989 2.051

Inorganic

fertilizer

5 t ha-1

0.223 0.613 0.735 0.827 1.073 1.189 1.164 1.181 1.325 1.419 1.487 1.513

7.5 t ha-1

0.120 0.318 1.069 1.166 1.532 1.795 1.882 1.903 2.016 2.054 2.102 2.117

10 t ha-1

0.218 0.412 1.012 1.124 1.799 1.921 2.156 2.166 2.185 2.204 2.217 2.279

Control Nil 0.032 0.155 0.223 0.504 1.612 1.935 1.065 1.409 1.510 1.532 1.544 1.451


1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.01 0.02 0.02 0.04 0.14 0.23 0.43 0.97 1.87 2.93 4.41 4.78

7.5 t ha-1

0.01 0.03 0.03 0.07 0.20 0.34 0.87 2.02 2.55 4.81 6.28 6.82

10 t ha-1

0.01 0.02 0.04 0.05 0.18 0.39 0.69 1.85 3.29 4.02 6.52 7.22

Inorganic

fertilizer

5 t ha-1

0.01 0.01 0.02 0.03 0.23 0.21 0.42 1.65 3.05 4.50 6.60 7.53

7.5 t ha-1

0.01 0.02 0.05 0.07 0.29 0.37 0.76 2.27 3.81 5.70 8.21 9.39

10 t ha-1

0.01 0.03 0.04 0.11 0.28 1.13 1.54 4.04 5.87 7.42 10.2 12.7

Control Nil 0.01 0.01 0.02 0.03 0.10 0.26 0.29 0.98 1.76 2.93 3.16 3.21

130




1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.357 0.747 1.687 2.193 2.661 3.034 3.052 3.122 3.201 3.254 3.388 3.438

7.5 t ha-1

0.467 0.826 1.807 1.953 2.409 2.947 3.047 3.16 3.229 3.395 3.474 3.545

10 t ha-1

0.546 1.275 2.124 2.581 2.722 3.167 3.226 2.559 3.688 3.667 3.86 3.917

Inorganic

fertilizer

5 t ha-1

0.611 1.157 1.727 2.011 2.507 2.852 2.93 2.964 3.144 3.245 3.324 3.353

7.5 t ha-1

0.488 1.073 2.152 2.335 3.009 3.565 3.844 3.962 4.112 4.169 4.25 4.353

10 t ha-1

0.786 1.259 2.056 2.296 3.477 4.001 4.335 4.345 4.382 4.41 4.442 4.570

Control Nil 0.135 0.62 0.863 1.411 2.740 3.099 2.242 2.602 2.711 2.756 2.782 2.767




1 2 3 4 5 6 7 8 9 10 11 12

Vermicompost

5 t ha-1

0.119 0.121 0.145 0.209 0.300 0.205 0.214 0.221 0.236 0.258 0.271 0.293

7.5 t ha-1

0.108 0.154 0.228 0.259 0.313 0.334 0.345 0.396 0.418 0.437 0.441 0.462

10 t ha-1

0.123 0.133 0.331 0.291 0.326 0.347 0.324 0.390 0.428 0.459 0.462 0.459

Inorganic

fertilizer

5 t ha-1

0.132 0.156 0.182 0.190 0.213 0.231 0.259 0.268 0.281 0.314 0.340 0.358

7.5 t ha-1

0.140 0.171 0.184 0.215 0.253 0.280 0.298 0.311 0.336 0.348 0.352 0.361

10 t ha-1

0.175 0.197 0.203 0.222 0.288 0.294 0.312 0.322 0.342 0.348 0.356 0.374

Control Nil 0.102 0.122 0.136 0.158 0.164 0.191 0.203 0.218 0.224 0.236 0.248 0.261

EFFECT OF VERMICAST GENERATED FROM ALLELOPATHIC

WEEDS AND PAPER WASTE ON PHYSICAL AND CHEMICAL

PROPERTIES OF POTTING SOIL GROWING CLUSTER BEAN

(CYAMOPSIS TETRAGONOLOBA)

Chapter

9

131



CChhaapptteerr 99

Effect of vermicast generated from allelopathic weeds and

paper waste on physical and chemical properties of potting

soil growing cluster bean (Cyamopsis tetragonoloba)

Abstract

There is accumulation of scientific evidence on the positive impact of vermicompost on plant growth.

However, the vermicompost derived from different parent materials have different physical, chemical and

biological qualities, and their impact on plant growth is also reported to vary considerably. To understand the

factors which attribute the differential impact of vermicompost from different parent materials on the growth

and yield of plants, it is necessary to recognize their impact on soil health. Hence, in the present study, the

application of vermicast generated from different organic wastes such as paper waste, leaves of ipomoea

(Ipomoea carnea), and of lantana (Lantana camara) on physical and chemical properties of potting soil

housing cluster bean (Cyamopsis tetragonoloba) were investigated. Seeds of cluster bean were sown in soil

to which vermicast was applied at the rate of 5, 7.5 and 10 t ha-1

respectively. In another set of treatment,

amounts of essential nutrients equivalent to those present in the vermicast treatments were applied in the

inorganic form and a control set comprising of only soil has been kept without any nutrient supplement.

Samples from all these treatments were collected on weekly basis during different stages of plant growth.

The essential physical and chemical variables of soil which is related to the plant growth were studied. The

results reveal that vermicast application created a suitable physical environment and also reduced the nutrient

loss at surface soil than the inorganic fertilizer.

1. Introduction

Vermicompost has long been recognized as

beneficial organic soil amendment in agriculture for

the maintenance of soil fertility to support plant

growth. Supplement of vermicompost, stimulate

seed germination, vegetative growth, shoot and root

development of several plant species. Stimulation

of flowering, increase in fruit yield and its

nutritional quality are also reported with

vermicompost supplement (Edwards et al., 2011).

Additionally, it increases the disease resistance of

plants to parasitic nematodes, insects, and mites

(Yardim et al., 2006; Zaller, 2006; Edwards et al.,

2009; Simsek-Ersahin et al., 2009; Simsek-Ersahin,

2011). The improvement in plant production and

protection with vermicompost substitution could be

attributed to several factors. First, vermicompost

amendments improve the overall physico-chemical

and biological characteristics of the soils that favor

better plant growth. The presence of plant growth-

influencing substances, such as plant growth

regulating hormones and humic acids in

vermicompost has also been suggested as a possible

factor contributing to increased plant growth and

yields (Tomati et al., 1988; Muscolo et al., 1999;

Arancon et al., 2003a; Arancon et al., 2006).

Through the vermicomposting technology,

wide range of organic wastes can be processed,

which includes paper waste (Elvira et al., 1997,

1998; Gajalakshmi and Abbasi, 2004), sewage

sludge (Domínguez et al., 2000; Khwairakpam and

Bhargava, 2009; Hait and Tare, 2011), urban

132

residues, and food and animal waste (Edwards et

al., 1985; Edwards, 1988; Domínguez and

Edwards, 1997; Atiyeh et al., 2001; Aira et al.,

2006; Garg et al., 2006; Suthar, 2007; Lazcano et

al., 2008), as well as horticultural residues

(Gajalakshmi et al., 2005; Gupta et al., 2007;

Pramanik et al., 2007; Suthar, 2007) and food

industry waste (Edwards, 1983; Butt, 1993;

Nogales et al., 1999a, 1999b, 2005). Nevertheless,

the vermicompost prepared from different organic

wastes, with different processes and duration,

produce a final product which differs in its quality

(Tandon, 1995; Campitelli and Ceppi, 2008).

The previous studies (Chapters 6, 7 and 8)

dealing with the effect of vermicast generated from

paper waste and from pernicious weed, lantana

(Lantana camara) and ipomoea (Ipomoea carnea)

on the germination, growth and yield of cluster

bean also manifests this finding. The vermicompost

generated from different parent material show

different physical and chemical qualities and their

influence on germination, growth and yield of test

plant also varied considerably. It was seen that the

application vermicompost derived from phytomass

such as lantana and ipomoea supported the

germination, growth and better fruit yield than

inorganic fertilizers. In contrast, the plants treated

with paper waste vermicompost exhibited lower

growth and yield than the equivalent inorganic

fertilizer. To understand the factors which attribute

the disparity on the growth and yields of plants

which were treated with vermicompost from

different parent materials, it is necessary to

understand their impact on soil health. Hence, the

objectives of the presence study are, (i) to evaluate

the impact of vermicast from different parent

materials on the physical and chemical properties of

potting soil germinating and growing cluster bean,

(ii) to determine the dose dependent changes in the

soil properties with vermicast application, and (iii)

to compare the impact of different vermicompost

on soil properties with soil fortified with inorganic

fertilizers (IF) in concentrations equivalent to those

present in the respective vermicompost (VC)

treatments.


The studies were conducted at Pondicherry

University, Puducherry, India, located on the east

coast of the Indian peninsula (11°56’N, 79°53’E).

This region experiences hot summers during March

– July (maximum day temperature 35-38°C), and

mild winters during December - February

(maximum day temperature 29-32°C). The average

annual rainfall is about 1300 mm, concentrated

mainly during October – December but with a few

rainy days occurring in July–August and January as

well. For vermicomposting, reactors fabricated with

aluminum sheet of 140 liter volume were employed

for direct vermicomposting of lantana and ipomoea

leaves by a process recently developed by the

author’s group (Gajalakshmi et al., 2002, 2005;

Abbasi et al., 2009, 2011; Kumar et al., 2012;

Tauseef et al., 2013). As paper waste is almost

entirely cellulosic, with only traces of elements

other than C, H, and O, the feed was spiked with

9% w/w of cow dung in order to provide NPK and

other nutrients in adequate amounts and

vermicomposted in similar fashion (Gajalakshmi et

al., 2012). The periodically harvested vermicast

was stored in sealed plastic containers. The study

on germination and growth were conducted

outdoors with 49 liter containers (40 cm height with

surface area 35 x 35 cm), lined with high-density

polyethylene (HDPE) sheets. The soil used in the

experiments was collected from a previously

uncultivated piece of land so that the results are not

influenced by any earlier fertilizer application. It is

sandy loam soil which is known to be low on

organic carbon and nutrients. The physico-chemical

properties of the vermicast and of the soil with

which it was used are given in Table 1. The

experiments were carried out during July – October

which is reported to be a season ideal for growing

cluster beans in the study area (ICAR, 2011).

133

Table 1. Chemical and physical properties of vermicast and soil used in the study.


For studying the impact of vermicast from

paper waste and leaves of lantana and ipomoea on

potting soil housing cluster bean, three sets of

experiments were conducted for each substrate. In

set I, one batch of containers was supplied with 7.5

t ha-1

of VC while two other batches were given 5

and 10 t ha-1

of VC. Set II comprised of chemical

fertilizers (IF) which were treated with nutrients N,

P, K, Ca, Mg, S, Fe, Mn, Cu, Zn, B, Mo and Cl in

concentrations equivalent to those present in the

respective vermicompost treatments. The last set

had no supplementation of nutrients and served as

control. Seven set, each comprising of 36

containers, or 252 containers in all, were prepared

in this fashion. The Pusanavabahar variety seeds of

the cluster bean, which is locally available in the

authors’ study area, was sown in all the containers.

Following germination, the plants were grown for

three months. Throughout the experiments,

adequate watering was done. Deweeding was done

periodically.

The soil samples from all the treatments were

collected on weekly basis during different stage of

plant growth, for physical and chemical analysis.

Undisturbed soil cores (5.7 cm diameter and 40 cm

Variables Vermicast

Lantana camara Ipomoea carnea Paper waste Soil

Chemical properties

pH 6.47±0.01 6.51±0.06 7.83±0.05 6.30±0.10


330.3±4.3 356.8±12.1 259.6±8.2 8.87±0.02


18.4±0.1 19.8±0.2 11.7±0.4 2.66±0.02


Phosphorus mg kg -1

80.7±0.9 37.83±0.60 71.40±3.31 0.41±0.01

Potassium g kg-1

5.73±0.03 2.12±0.04 1.98±0.10 0.40±0.00

Sulphur mg 100g-1

1.34±0.01 4.64±0.09 0.40±0.04 0.54±0.01

Calcium g kg-1

5.49±0.02 15.5±2.6 15.9±1.4 8.27±0.01

Magnesium g kg-1

5.77±0.05 4.85±0.09 7.13±0.07 0.09±0.02

Boron mg kg-1

46.2±0.8 78.1±2.3 7.28±0.09 26.9±1.2

Copper mg kg-1

44.2±3.9 10.6±0.3 16.9±1.0 5.08±0.15

Iron mg kg-1

50.6±2.0 270.3±14.7 208.3±11.4 59.9±2.9

Manganese mg kg-1

227.2±8.4 117.4±5.0 63.8±1.8 45.1±2.2

Zinc mg kg-1

162.4±6.0 107.0±7.7 94.1±2.2 55.0±2.6

Molybdenum mg kg-1

BDL BDL BDL BDL

Physical properties

Dry weight % 43.1±0.3 48.4±1.1 49.6±1.9 94.7±0.1

Bulk density g cm-3

0.40±0.00 0.26±0.02 0.24±0.01 1.28±0.00


1.35±0.01 1.30±0.05 1.21±0.03 2.70±0.12

Water-holding capacity % 248.0±10.2 261.9±16.6 118.0±11.2 36.9±2.4

Electrical conductivity mmhos cm-1

9.36±0.01 6.25±0.07 2.96±0.04 0.12±0.02

Total porosity % 70.6±0.2 80.2±1.8 80.08±1.0 53.1±2.0

Air filled porosity % 48.1±0.1 66.8±2.6 68.14±1.2 46.5±2.1

134

height) were taken from the 0 to 30 cm layer by

inserting core sampler gently into the soil. The

sampled cores were divided into two pieces (0-15

cm and 15-30 cm) and packed separately in airtight

polyethylene bags. These samples were stored at

2°C until analyses could take place. Forty two soil

samples per week (7 treatments x 3 replications x 2

soil depth layers: 0–15 cm, 15–30 cm), leading to

504 samples overall were collected during the

course of three months of the experiment. The

samples were air-dried at room temperature,

crushed, and passed through a 2 mm diameter

sieve, before analyses. For bulk density

measurement, undisturbed soil cores were collected

separately from each soil depth.

Total organic carbon (Corg) was measured by

modified dichromate redox method according to

Heanes (1984). Total nitrogen (Ntot) content was

determined by the modified Kjeldahl method


digester and distillation units. Inorganic N - NH4+

and NO3- were extracted in 2M KCl solution (1:10,

w/v) and determined by modified indophenol blue

and Devarda’s alloy methods, respectively (Jones

2001; Bashour and Sayegh, 2007). Extractable

potassium (Kext) and calcium (Caext) were

determined using a flame photometer (Elico™

CL378) after extraction with neutral 1N ammonium

acetate solution (Carter and Gregorich, 2008).

Extractable form of magnesium (Mgext), boron

(Bext), copper (Cuext), iron (Feext), manganese

(Mnext), zinc (Znext), molybdenum (Moext) were

determined using a Jobin Yvon – Ultima 2 model

inductively coupled plasma atomic emission

spectroscopy (ICP-AES) by extracting

sample/solution ratio of 1:25 (w/v) with Mehlich 3

extraction solution (Mehlich, 1984). The same

extract was used to determine the extractable

phosphorus (Pext) according to the ammonium

molybdate-ascorbic acid method (Knudsen and

Beegle, 1988).

The pH and electrical conductivity (EC) of

the samples were measured in suspension of 1:2

(v/w) by using EI™ 611E EC meter and Digison™

digital pH meter 7007 respectively. Bulk density

and particle density were measured by undisturbed

core and volumetric flask methods, respectively

(Bashour and Sayegh, 2007). The total and water-

filled porosity (WFPS) were calculated from the

particle and bulk density values of respective

samples using standard formulae (Carter and

Gregorich, 2008). Water-holding capacity (WHC)

of the samples was obtained by determining their

water retention ability after they were immersed in

water and the excess water was drained off

(Margesin and Schinner, 2005).



difference between the results. SPSS version 16

was used for this purpose.


3.1. Physical properties

The physical properties of soil were

significantly affected by different VC and IF

treatments (p<0.05; Tables 2-8). The soil treated

with VC from all three substrates had significantly

lower bulk density than the ones treated with

corresponding IF treatments and control (Table 2).

During the initial month, there was no significant

difference in bulk density of soils treated with VC

and the equivalent IF. However, in the second and

third months, the bulk density of soil treated with

ipomoea based VC was about 7% lower than the IF

treatments and those of lantana and paper waste VC

showed 8 and 12% lesser bulk density, respectively.

The bulk density of lantana based VC treated soil

were 23% lower than the control; whereas the

ipomoea and paper waste VC treatments showed 20

and 16% lesser bulk density, respectively. In all the

VC treatments, increasing dose of VC showed a

steady decline in bulk density, which was possibly

due to the reasons, (i) addition of lower density

organic matter in the form of vermicast might have

diluted the bulk density of mineral soil (Tejada et

135

Table 2. Changes in the bulk density (g cm−3

) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent

inorganic fertilizers (IF), at different periods of time (mean ± SD).

Treatments Lantana Ipomoea Paper waste

July Aug Sep July Aug Sep July Aug Sep

Layer

0-15 cm VC 1.08±0.06 1.13±0.04 1.15±0.04 1.06±0.11 1.14±0.05 1.19±0.05 1.11±0.05 1.17±0.04 1.23±0.03

IF 1.07±0.07 1.18±0.04 1.25±0.03 1.10±0.08 1.22±0.06 1.27±0.04 1.10±0.05 1.17±0.03 1.21±0.02

Cont. 1.23±0.04 1.27±0.01 1.29±0.01 1.26±0.04 1.30±0.01 1.32±0.01 1.26±0.04 1.30±0.01 1.32±0.01

15-30 cm VC 1.05±0.06 1.10±0.05 1.13±0.05 1.03±0.07 1.12±0.06 1.17±0.05 1.08±0.04 1.15±0.04 1.22±0.03

IF 1.06±0.08 1.16±0.04 1.23±0.03 1.07±0.08 1.18±0.05 1.24±0.05 1.09±0.06 1.16±0.04 1.20±0.02

Cont. 1.20±0.03 1.23±0.01 1.27±0.03 1.23±0.03 1.26±0.01 1.30±0.03 1.23±0.03 1.26±0.01 1.30±0.03

F value 6.517***

22.59***

3.847**

5.248**

12.22***

16.49***

9.309***

17.44***

25.04***

Amount

5 t ha-1

VC 1.07±0.06 1.15±0.01 1.18±0.01 1.13±0.02 1.18±0.03 1.25±0.02 1.14±0.04 1.20±0.02 1.25±0.03

IF 1.06±0.05 1.19±0.03 1.26±0.02 1.15±0.08 1.27±0.03 1.31±0.01 1.07±0.05 1.16±0.01 1.21±0.01

7.5 t ha-1

VC 1.12±0.02 1.14±0.01 1.16±0.01 0.94±0.05 1.08±0.05 1.15±0.03 1.09±0.02 1.16±0.02 1.22±0.02

IF 0.99±0.05 1.13±0.04 1.21±0.02 1.01±0.06 1.15±0.02 1.22±0.04 1.07±0.03 1.14±0.03 1.19±0.01

10 t ha-1

VC 1.01±0.04 1.05±0.02 1.08±0.02 1.06±0.07 1.13±0.01 1.14±0.01 1.05±0.03 1.12±0.02 1.20±0.03

IF 1.14±0.03 1.20±0.03 1.25±0.03 1.09±0.03 1.19±0.03 1.24±0.02 1.15±0.03 1.19±0.01 1.23±0.01

Control

1.21±0.04 1.25±0.03 1.28±0.02 1.24±0.04 1.28±0.03 1.31±0.02 1.24±0.04 1.28±0.03 1.31±0.02

F value

21.09***

68.46***

4.212**

20.50***

45.52***

82.83***

21.99***

47.96***

44.18***

*p<0.05,

**p<0.01,

***p<0.001,

n.s. not significant

136

Table 3. Changes in the particle density (g cm−3





Layer

0-15 cm VC 2.63±0.03 2.64±0.07 2.70±0.30 2.65±0.04 2.69±0.04 2.68±0.05 2.61±0.04 2.62±0.05 2.65±0.04

IF 2.65±0.09 2.65±0.02 2.63±0.03 2.66±0.05 2.66±0.03 2.70±0.08 2.65±0.03 2.67±0.06 2.66±0.02

Cont. 2.64±0.02 2.65±0.05 2.66±0.03 2.64±0.02 2.65±0.05 2.66±0.03 2.64±0.02 2.65±0.05 2.66±0.03

15-30 cm VC 2.62±0.04 2.64±0.04 2.67±0.04 2.63±0.02 2.65±0.03 2.65±0.03 2.63±0.05 2.63±0.07 2.63±0.05

IF 2.64±0.05 2.65±0.03 2.92±1.03 2.67±0.08 2.65±0.03 2.64±0.08 2.66±0.02 2.66±0.04 2.65±0.04

Cont. 2.64±0.02 2.68±0.04 2.64±0.03 2.64±0.02 2.68±0.04 2.64±0.03 2.64±0.02 2.68±0.04 2.64±0.03

F value 0.401n.s

0.748n.s

0.637n.s

0.597n.s

1.990n.s

1.738n.s

3.183* 1.589

n.s 0.998

n.s

Amount

5 t ha-1 VC 2.62±0.03 2.62±0.07 2.61±0.11 2.63±0.03 2.67±0.02 2.67±0.05 2.58±0.04 2.63±0.08 2.60±0.04

IF 2.66±0.05 2.64±0.02 2.63±0.03 2.68±0.04 2.67±0.04 2.67±0.05 2.66±0.01 2.70±0.04 2.67±0.03

7.5 t ha-1 VC 2.63±0.02 2.65±0.04 2.75±0.34 2.65±0.04 2.66±0.05 2.65±0.06 2.62±0.03 2.61±0.03 2.66±0.04

IF 2.60±0.05 2.65±0.03 2.66±0.04 2.68±0.10 2.64±0.02 2.67±0.07 2.66±0.03 2.64±0.06 2.64±0.02

10 t ha-1 VC 2.61±0.04 2.65±0.04 2.68±0.05 2.64±0.03 2.68±0.04 2.66±0.04 2.65±0.04 2.64±0.06 2.66±0.03

IF 2.67±0.08 2.66±0.04 3.04±1.26 2.64±0.04 2.64±0.03 2.67±0.12 2.65±0.02 2.66±0.04 2.65±0.03

Control

2.64±0.02 2.67±0.05 2.65±0.03 2.64±0.02 2.67±0.05 2.65±0.03 2.64±0.02 2.67±0.05 2.65±0.03

F value

1.852n.s

1.149n.s

0.891n.s

0.924n.s

1.228n.s

0.190n.s

5.736***

2.363* 5.185

***

*p<0.05,

**p<0.01,

***p<0.001,


137

Table 4. Changes in the total porosity (%) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent




Layer

0-15 cm VC 59.0±2.2 57.2±2.3 56.9±4.1 60.0±4.5 57.4±2.0 55.5±2.1 57.6±2.4 55.3±2.1 53.5±1.6

IF 59.6±1.9 55.4±1.8 52.7±1.7 58.8±2.9 54.2±1.8 52.9±2.1 58.5±2.0 56.2±1.2 54.5±0.7

Cont. 53.6±1.6 52.1±1.0 51.4±0.6 52.5±1.6 50.9±1.0 50.3±0.6 52.5±1.6 50.9±1.0 50.3±0.6

15-30 cm VC 59.9±2.2 58.2±1.9 57.7±2.3 60.9±2.6 58.0±2.1 55.8±2.0 58.8±2.0 56.3±1.8 53.8±1.8

IF 60.0±3.2 56.1±1.4 55.3±7.2 59.8±3.5 55.4±2.0 53.1±2.3 59.2±2.3 56.5±1.5 54.6±1.0

Cont. 54.7±1.1 54.2±1.0 51.9±1.4 53.6±1.1 53.1±1.0 50.7±1.4 53.6±1.1 53.1±1.0 50.7±1.4

F value 7.339***

15.83***

2.042n.s

4.807**

13.63***

11.51***

7.605***

10.33***

14.56***

Amount

5 t ha-1

VC 59.1±1.9 56.0±1.5 54.8±2.2 57.1±1.3 55.6±1.2 53.2±1.2 55.8±1.6 54.1±1.7 51.9±1.3

IF 60.3±2.1 55.0±1.0 52.1±1.1 57.2±2.7 52.6±1.1 51.1±0.9 59.9±2.0 57.1±0.4 54.8±0.7

7.5 t ha-1

VC 57.7±0.8 56.9±0.6 57.4±3.9 64.5±2.0 59.6±1.6 56.7±1.0 58.3±0.6 55.7±0.7 54.1±1.1

IF 61.8±2.0 57.2±1.6 54.5±1.3 62.2±3.0 56.5±1.0 54.4±1.9 59.9±1.3 56.8±1.6 55.1±0.6

10 t ha-1

VC 61.5±1.6 60.3±0.6 59.7±1.4 59.7±2.4 58.0±0.8 57.1±0.5 60.4±1.3 57.7±1.4 55.0±1.0

IF 57.3±1.0 55.0±1.4 55.4±9.1 58.5±1.1 55.2±1.0 53.6±2.1 56.7±1.1 55.2±0.8 53.7±0.5

Control

54.2±1.4 53.1±1.4 51.6±1.0 53.1±1.4 52.0±1.5 50.5±1.0 53.1±1.4 52.0±1.5 50.5±1.0

F value

19.01***

37.78***

2.330* 19.21

*** 41.34

*** 34.38

*** 23.06

*** 20.25

*** 36.52

***

*p<0.05,

**p<0.01,

***p<0.001,


138

Table 5. Changes in the water-filled pore space (%) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or

equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).



Layer

0-15 cm VC 24.7±4.7 25.3±5.3 30.2±4.3 20.0±5.4 26.4±3.5 27.4±3.9 24.0±3.4 26.0±4.2 28.9±4.7

IF 23.3±2.5 27.3±5.5 31.6±7.6 25.4±6.2 29.8±6.0 29.1±5.8 22.9±4.5 25.7±4.9 27.6±3.9

Cont. 26.1±1.7 28.5±5.2 29.1±3.5 27.3±1.8 29.9±5.5 30.5±3.7 27.3±1.8 29.9±5.5 30.5±3.7

15-30 cm VC 26.8±3.7 30.5±4.3 29.2±4.6 21.2±5.1 25.9±4.3 32.4±5.9 27.2±3.9 29.5±4.7 33.0±4.8

IF 27.6±3.0 31.2±5.3 33.7±5.0 25.8±4.9 30.7±6.5 33.5±5.0 25.9±3.3 28.4±4.2 31.1±3.6

Cont. 29.2±1.2 30.6±3.2 33.0±2.1 30.5±1.2 32.1±3.3 34.5±2.3 30.5±1.2 32.1±3.3 34.5±2.3

F value 2.891* 2.865

* 2.103

n.s 3.168

* 2.060

n.s 3.542

** 2.987

* 2.115

n.s 4.019

**

Amount

5 t ha-1

VC 25.9±5.1 30.2±4.6 31.3±4.2 24.7±2.9 26.4±4.7 31.8±5.8 26.6±5.3 30.9±4.1 33.3±5.1

IF 24.7±3.2 31.5±7.5 37.0±7.2 27.1±4.8 33.3±6.5 32.9±4.9 24.1±5.0 24.4±4.7 30.3±4.1

7.5 t ha-1

VC 27.3±3.7 30.1±3.4 29.5±5.8 16.0±2.4 24.5±3.4 28.5±5.7 25.6±3.2 27.7±4.4 31.2±5.6

IF 24.7±3.3 26.5±4.2 30.6±5.3 20.4±2.2 25.1±4.0 30.5±5.1 22.8±3.1 28.6±2.7 27.6±4.3

10 t ha-1

VC 24.1±4.0 23.4±5.5 28.3±2.4 21.2±5.4 27.5±3.3 29.5±5.0 24.5±3.3 24.7±3.7 28.3±3.4

IF 27.1±3.9 29.8±3.8 30.4±4.6 29.4±4.5 32.3±4.4 30.6±7.3 26.2±4.0 28.1±5.5 30.2±3.8

Control

27.6±2.1 29.6±4.1 31.1±3.4 28.9±2.2 31.0±4.3 32.5±3.6 28.9±2.2 31.0±4.3 32.5±3.6

F value

1.156n.s

3.053* 1.073

n.s 10.87

*** 5.069

*** 0.880

n.s 1.565

n.s 3.046

* 2.302

*

*p<0.05,

**p<0.01,

***p<0.001,


139

Table 6. Changes in the water holding capacity (%) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or




Layer

0-15 cm VC 41.1±4.2 43.9±3.5 46.4±3.2 45.4±5.1 48.6±6.0 50.6±6.6 42.2±5.0 45.1±5.0 48.1±5.6

IF 33.5±1.6 35.3±0.9 36.4±0.9 35.2±2.8 38.6±2.8 39.7±2.6 36.5±1.6 39.3±3.0 42.6±2.3

Cont. 31.0±0.6 33.4±0.6 35.5±0.8 31.0±0.6 33.4±0.6 35.5±0.8 31.0±0.6 33.4±0.6 35.5±0.8

15-30 cm VC 39.0±2.6 41.7±2.5 44.3±2.6 43.3±5.3 46.6±6.0 49.8±6.3 39.2±4.4 42.1±4.1 44.9±4.0

IF 32.1±1.5 33.3±0.9 34.5±0.7 35.0±1.6 37.5±1.3 39.9±1.1 35.3±1.3 37.7±1.3 40.5±1.8

Cont. 32.9±0.2 33.8±1.0 36.3±0.7 32.9±0.2 33.8±1.0 36.3±0.7 32.9±0.2 33.8±1.0 36.3±0.7

F value 18.87***

46.53***

83.67***

15.45***

18.71***

23.31***

8.784***

12.99***

16.94***

Amount

5 t ha-1

VC 36.5±0.8 39.5±1.7 41.7±1.1 38.3±1.0 39.8±1.2 41.5±0.5 34.9±1.3 37.7±1.5 40.1±1.0

IF 32.6±1.7 34.1±1.6 35.4±1.2 36.4±1.1 38.7±1.9 40.4±0.8 34.8±0.7 36.7±0.9 39.6±1.4

7.5 t ha-1

VC 39.9±1.8 42.8±1.1 46.3±1.1 45.0±2.7 50.3±2.5 53.5±0.7 42.0±2.3 45.5±2.0 49.1±2.7

IF 34.3±1.2 35.1±1.3 36.2±1.3 36.3±1.2 39.1±2.2 41.2±1.3 37.1±1.2 39.4±2.2 42.8±0.7

10 t ha-1

VC 43.8±2.6 46.1±2.2 48.1±1.9 49.7±2.2 52.7±1.5 55.5±1.6 45.2±2.3 47.5±2.3 50.2±2.3

IF 31.5±0.7 33.9±1.1 34.9±1.0 32.6±1.6 36.3±1.8 37.8±1.7 35.8±1.6 39.4±2.8 42.3±2.7

Control

31.9±1.1 33.6±0.8 35.9±0.8 31.9±1.1 33.6±0.8 35.9±0.8 31.9±1.1 33.6±0.8 35.9±0.8

F value

54.93***

95.63***

207.6***

91.47***

132.8***

443.9***

51.31***

51.92***

78.60***

*p<0.05,

**p<0.01,

***p<0.001,


140

Table 7. Changes in the electrical conductivity (mmhos cm−1

) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste,

or equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).



Layer

0-15 cm VC 0.18±0.03 0.20±0.02 0.19±0.02 0.17±0.02 0.18±0.02 0.18±0.01 0.15±0.02 0.16±0.02 0.15±0.02

IF 0.19±0.02 0.19±0.02 0.17±0.02 0.19±0.02 0.19±0.02 0.17±0.02 0.17±0.02 0.17±0.01 0.16±0.00

Cont. 0.11±0.00 0.12±0.00 0.12±0.00 0.11±0.00 0.12±0.00 0.12±0.00 0.11±0.00 0.12±0.00 0.12±0.00

15-30 cm VC 0.18±0.01 0.19±0.02 0.18±0.01 0.17±0.01 0.18±0.01 0.18±0.01 0.14±0.01 0.14±0.02 0.13±0.02

IF 0.20±0.03 0.20±0.03 0.18±0.02 0.20±0.03 0.19±0.02 0.18±0.02 0.18±0.02 0.18±0.02 0.17±0.02

Cont. 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00

F value 15.59***

21.57***

32.74***

23.29***

28.97***

50.93***

20.46***

19.33***

29.00***

Amount

5 t ha-1

VC 0.16±0.00 0.17±0.00 0.17±0.00 0.16±0.01 0.17±0.00 0.17±0.00 0.14±0.01 0.14±0.01 0.12±0.01

IF 0.17±0.01 0.16±0.01 0.15±0.01 0.17±0.01 0.16±0.00 0.16±0.01 0.17±0.01 0.17±0.01 0.16±0.00

7.5 t ha-1

VC 0.17±0.01 0.20±0.01 0.19±0.01 0.17±0.01 0.19±0.01 0.19±0.00 0.14±0.01 0.14±0.01 0.13±0.01

IF 0.20±0.01 0.20±0.01 0.18±0.01 0.20±0.01 0.20±0.01 0.18±0.01 0.16±0.01 0.16±0.01 0.16±0.00

10 t ha-1

VC 0.20±0.02 0.21±0.01 0.20±0.01 0.18±0.01 0.19±0.01 0.19±0.00 0.17±0.01 0.18±0.01 0.16±0.01

IF 0.22±0.01 0.22±0.01 0.20±0.00 0.22±0.01 0.21±0.01 0.19±0.01 0.19±0.01 0.20±0.02 0.18±0.02

Control

0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00 0.12±0.00

F value

58.90***

102.3***

227.6***

77.90***

184.5***

343.5***

62.71***

53.14***

56.16***

*p<0.05,

**p<0.01,

***p<0.001,


141

Table 8. Changes in the pH of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or equivalent inorganic fertilizers

(IF), at different periods of time (mean ± SD).

Treatments

Lantana Ipomoea Paper waste


Layer

0-15 cm VC 6.22±0.07 6.17±0.09 6.15±0.11 6.30±0.01 6.31±0.03 6.30±0.03 6.40±0.04 6.70±0.28 6.82±0.29

IF 6.24±0.06 6.22±0.06 6.21±0.06 6.52±0.29 6.61±0.41 6.61±0.39 6.83±0.53 7.03±0.48 7.07±0.46

Cont. 6.30±0.00 6.28±0.01 6.27±0.00 6.30±0.00 6.28±0.01 6.27±0.00 6.30±0.00 6.28±0.01 6.27±0.00

15-30 cm VC 6.06±0.08 5.93±0.29 5.98±0.01 6.10±0.13 6.08±0.13 6.07±0.13 6.28±0.08 6.33±0.08 6.36±0.08

IF 6.09±0.08 6.04±0.05 6.02±0.04 6.16±0.10 6.17±0.12 6.18±0.13 6.34±0.08 6.40±0.09 6.42±0.08

Cont. 6.27±0.02 6.21±0.01 6.19±0.01 6.27±0.02 6.21±0.01 6.19±0.01 6.27±0.02 6.21±0.01 6.19±0.01

F value 15.57***

9.140***

41.59***

9.745***

0.901n.s

14.71***

7.187***

17.03***

1.199n.s

Amount

5 t ha-1

VC 6.11±0.14 6.09±0.14 6.09±0.13 6.28±0.02 6.28±0.02 6.27±0.02 6.29±0.09 6.36±0.13 6.40±0.14

IF 6.19±0.12 6.17±0.12 6.16±0.12 6.28±0.03 6.27±0.03 6.27±0.02 6.34±0.07 6.41±0.11 6.44±0.11

7.5 t ha-1

VC 6.22±0.08 6.01±0.41 6.11±0.13 6.16±0.15 6.14±0.15 6.13±0.15 6.33±0.11 6.45±0.14 6.54±0.20

IF 6.22±0.06 6.17±0.08 6.14±0.10 6.19±0.18 6.19±0.20 6.20±0.20 6.45±0.15 6.68±0.33 6.72±0.34

10 t ha-1

VC 6.10±0.06 6.05±0.03 5.99±0.02 6.16±0.17 6.16±0.19 6.16±0.19 6.40±0.03 6.74±0.35 6.83±0.40

IF 6.08±0.09 6.06±0.09 6.05±0.09 6.55±0.40 6.71±0.48 6.72±0.45 6.97±0.61 7.06±0.59 7.08±0.58

Control

6.28±0.02 6.24±0.04 6.23±0.04 6.28±0.02 6.24±0.04 6.23±0.04 6.28±0.02 6.24±0.04 6.23±0.04

F value

4.572**

1.754n.s

6.263***

3.162* 0.997

n.s 9.221

*** 5.883

*** 7.027

*** 1.044

n.s

*p<0.05,

**p<0.01,

***p<0.001,


142

al., 2009), and (ii) vermicast containing binding

agents such as polysaccharides and bacterial gums

would have promoted the soil aggregation, which

might have increased the porosity of soil resulting

in reduction in bulk density (Bhatia and Shukla,

1982; Rasool et al., 2007). In the case of IF treated

soil, the highest reduction in bulk density was

recorded in 7.5 t ha-1

VC equivalent IF treatments,

followed by 10 and 5 t ha-1

. In most of the cases,

bulk density of the IF treated soil were 10% lesser

than the control. Few past studies also showed that

the balanced inorganic fertilizer application

increased the porosity of soil, resulting in the

reduction of bulk density (Yeoh and Oades, 1981;

Schjonning et al., 1994; Prasad and Sinha, 2000).

In all the treatments, the first 15 cm soil layer had

higher bulk density than deeper soil layer (15-30

cm). The particle density of untreated soil was

2.70±0.12 g cm-3

, which has not changed

considerably throughout the experiment (Table 3).

Total porosity of ipomoea and lantana based

VC treated soil were about 5% higher than the

corresponding IF treatments; whereas the paper

waste VC treated soil had shown 2% lower porosity

than the IF treatments (Table 4). Lantana and

ipomoea VC treated soil showed about 10% higher

porosity compared to the control treatment. Paper

waste based VC treatments increased the porosity

to the extent of 6%. In all IF treatments, porosity of

soil increased to the extent of 7% in comparison

with control. The VC and balanced fertilizer

application might have improved the aggregation

by microbial extracellular polysaccharide

production, filamentous structures (e.g. fungal

hyphae) and extruded biopolymer-induced

aggregation (Oades, 1993), resulting in an increase

in porosity of soil (Bhatia and Shukla, 1982).

Moreover, the VC amendment may have enhanced

the microbial decomposition of organic matter it

contained, and subsequent gaseous release fractured

the soil matrix which might have increased the pore

space (Pagliai et al., 1981).

The water-filled pore space (WFPS) of soils

has not showed any significant variation with

different treatments (Table 5). In most of the cases,

WFPS of soil treated with VC and IF were similar

to the control. The different doses of VC and IF

applications have also not had any significant

difference in WFPS. The water-holding capacity

(WHC) of soil significantly increased with

increasing dose of VC application (Table 6). Higher

WHC of VC treated soils may be probably the

reason of high pore space in these treatments (Liu

et al., 2013). Amongst different VC treatments, the

ipomoea VC showed higher increase of about 28%

in WHC compared to the control. In both lantana

and paper waste VC; it increased to the extent of

21%. The IF treatments increased the WHC to the

extent of 10% in comparison to the control. In all

the treatments, increase in WHC was prominent in

the first 15 cm layer than the soil below. EC of both

VC and IF treated soil were higher than the control

(Table 7). In the lantana and ipomoea based VC

and equivalent IF treatments, EC increased to the

extent of 39%, and in the paper waste VC and

equivalent IF treatments, it increased to the extent

of 15 and 31%, respectively in comparison to the

control. During the first month, higher EC was

recorded in both VC and IF treatments, and then

declined till the end. There was no significant

difference in EC of soil either treated with different

VC and IF, or collected from different layers. The

pH of soil from all the treatments was in the range

of 5.95 – 7.64. All IF treatments showed slightly

higher pH than the corresponding VC treatments

(Table 8).

3.2. Chemical properties

Chemical characteristics of soil were

significantly influenced by different treatments

(p<0.001; Tables 9-19). Application of VC

significantly increased the organic carbon (Corg)

content in soil; more than 50% higher compared to

the control and IF treatments during the first month

(Table 9). Among different treatments, the lantana

143

based VC increased the Corg maximally, followed

by ipomoea and paper waste VC. The Corg content

of IF treated soil did not show any significant

difference compared to the control. However,

throughout the experiment the Corg in all the

treatments increased, which may be due to the

addition of fallen leaves from the experimental

plants. The VC treated soils showed higher Corg

content throughout which may be due to

stabilization of humified organic compounds in the

vermicast as organo-mineral complex in soil (Li et

al., 2000). Total nitrogen (Ntot) of experimental soil

was higher after the fertilization with VC and IF

(Table 10). In the case of IF treated soils, higher

Ntot was in 15-30 cm layer from the surface than the

first 15 cm layer. The VC treated soil had higher

Ntot in surface soil layer during the first two months

and then the deeper soil layer showed high Ntot

content. The slow release of nutrients by the VC

might have caused higher Ntot retention in surface

soil layer during first two months. Later, increase of

Ntot in deeper soil layers of VC treatments may be

probably due to symbiotic nitrogen fixation by

rhizobia in the roots of the leguminous

experimental plants. These findings are also

supported by higher stimulation of nodulation in

plants grown in experimental soil with VC than

with IF and control.

The ammonium (NH4+) and nitrate (NO3

-)

content of soil were also significantly higher in VC

treatments than the IF treatments and control

(p<0.001; Tables 11-12). Ipomoea based VC

increased the soil NH4+ to the extent of 91% than

the control; and those of lantana and paper waste

based VC showed up to 88 and 74% increase,

respectively. The higher mineral nitrogen content in

VC treated soil was probably the reason of its

higher Corg content which could have provided a

larger source of N for mineralization. Hence, the

VC based treatments might have produced more

residual N in soil than the inorganic fertilizer

(Nethra et al., 1999; Arancon et al., 2006). As the

number of days progressed, the NH4+ content of the

soil reduced to 80% with all the three VC

treatments. In the case of IF treated soils, the NH4+

reduced to the extent of 87%. The NO3- content of

experimental soil also showed similar trend of

results with both VC and IF treatments. In both VC

and IF treatments, the NO3- content reduced up to

46% at the end of the experiment, and the reduction

was much pronounced in ipomoea based VC and its

equivalent IF treatments.

The lantana and ipomoea VC and its

equivalent IF application increased the soil

extractable phosphorus (Pext) content to the extent

of 10% and those of paper waste VC and IF showed

up to 3% (Table 13). As the days progressed, the

Pext content of soil reduced in all the treatments,

which was much pronounced in IF treatments. In all

the VC and IF treatments the higher Pext reduction

was recorded in 15-30 cm soil layer from the

surface than the first 15 cm layer. In the deeper soil

layer, the extensive root of the plants grown in

experimental soil might have uptaken the Pext

maximally resulting in higher Pext depletion in this

layer. The extractable potassium (Kext) content of

soil increased to the extent of 8% after fertilization

with both VC and IF (Table 14). As the days

progressed, the Kext content of soils showed

declining trend with all the treatments. At the end

of experiment, higher reduction in Kext content was

in IF treatments than the corresponding VC

treatments. Among the different VC treatments, the

lantana based VC showed higher reduction of up to

23% in Kext content and in its equivalent IF

treatments, it was up to 25% of reduction. The

reduction of Kext in the ipomoea and paper waste

based VC and its equivalent IF treatments were up

to a maximum of 20%.

Extractable calcium (Caext) content of soil

showed higher reduction to the extent of 45 and

50% with lantana based VC and its equivalent IF

treatments, respectively (Table 15). The paper

waste and ipomoea based treatments showed

maximum reduction of 45%. In all the VC

treatments, the first 15 cm layer of soils showed

higher Caext content than the corresponding IF

144

Table 9. Changes in the organic carbon (g kg−1





Layer

0-15 cm VC 25.0±4.6 27.1±4.9 32.7±6.0 23.5±5.7 27.2±4.3 30.0±5.1 18.0±3.3 21.7±4.0 24.8±4.5

IF 14.6±1.4 19.6±3.1 23.6±2.7 17.3±2.0 18.0±2.3 22.5±2.7 13.6±1.0 15.9±1.6 18.5±1.3

Cont. 16.8±2.0 17.5±1.4 18.7±1.1 16.8±2.0 17.5±1.4 18.7±1.1 16.8±2.0 17.5±1.4 18.7±1.1

15-30 cm VC 20.7±3.2 23.3±3.6 27.9±4.8 18.7±5.9 22.3±5.5 25.1±4.7 14.1±3.3 17.1±2.7 21.0±3.9

IF 13.9±1.8 16.2±2.0 20.4±2.0 13.5±1.4 16.2±1.3 22.0±2.8 13.1±2.2 13.5±2.0 16.5±1.3

Cont. 13.2±1.2 16.2±1.7 20.0±1.3 13.2±1.2 16.2±1.7 20.0±1.3 13.2±1.2 16.2±1.7 20.0±1.3

F value 27.71***

27.87***

25.89***

7.829***

20.33***

17.83***

8.617***

14.33***

15.89***

Amount

5 t ha-1

VC 18.8±2.2 20.8±2.5 24.2±3.1 16.2±2.4 21.6±2.1 23.6±1.6 14.2±4.0 15.7±2.4 18.4±2.1

IF 13.2±1.3 16.5±2.5 20.8±1.9 15.2±2.0 17.1±2.2 25.2±1.7 12.7±1.1 14.1±1.6 17.9±1.1

7.5 t ha-1

VC 23.5±2.5 26.4±2.3 32.8±3.2 18.9±4.0 21.9±5.0 25.5±4.5 15.8±3.3 19.8±2.7 23.0±2.5

IF 14.7±0.8 18.3±3.0 21.3±2.5 16.0±3.6 16.4±1.0 19.6±0.4 14.7±2.2 15.0±2.6 18.1±1.4

10 t ha-1

VC 26.3±4.6 28.4±4.8 33.9±5.3 28.3±3.2 30.8±2.1 33.7±2.8 18.2±3.5 22.7±3.9 27.4±3.3

IF 14.8±2.1 19.0±3.5 23.9±3.2 15.1±2.2 17.7±2.6 22.0±1.6 12.7±0.7 15.1±2.3 16.6±2.0

Control

15.0±2.4 16.8±1.6 19.4±1.3 15.0±2.4 16.8±1.6 19.4±1.3 15.0±2.4 16.8±1.6 19.4±1.3

F value

30.86***

38.29***

43.49***

19.36***

43.42***

53.47***

6.05***

15.57***

35.78***

*p<0.05,

**p<0.01,

***p<0.001,


145

Table 10. Changes in the total nitrogen (g kg−1





Layer

0-15 cm VC 7.4±4.0 7.6±4.9 3.8±2.3 7.5±3.7 5.8±3.4 3.3±1.2 4.3±1.7 3.9±1.7 2.7±0.7

IF 7.0±4.9 6.4±4.9 1.7±0.9 6.7±4.4 5.5±4.1 2.1±0.9 5.8±3.1 5.2±3.6 2.7±1.0

Cont. 0.4±0.1 0.7±0.2 0.5±0.2 0.4±0.1 0.7±0.2 0.5±0.2 0.4±0.1 0.7±0.2 0.5±0.2

15-30 cm VC 2.8±1.3 3.3±1.1 4.1±1.4 4.1±2.2 3.6±1.5 4.8±1.7 1.9±0.9 2.2±0.9 1.8±0.4

IF 5.9±3.2 5.4±2.9 2.7±0.9 5.8±2.8 4.0±2.7 2.2±0.9 4.3±2.4 3.6±2.3 2.3±1.2

Cont. 0.4±0.1 1.2±0.4 1.5±0.4 0.4±0.1 1.2±0.4 1.5±0.4 0.4±0.1 1.2±0.4 1.5±0.4

F value 5.954**

5.528**

11.47***

4.173** 4.072**

17.27***

8.311***

5.822**

8.263***

Amount

5 t ha-1

VC 2.8±1.2 3.0±0.7 2.5±0.6 3.6±1.2 2.7±0.7 2.7±1.0 1.9±0.7 2.0±0.7 1.6±0.3

IF 2.1±1.1 3.0±1.3 1.5±0.6 2.5±0.8 2.0±0.9 2.0±0.8 1.8±0.7 1.6±0.8 1.2±0.3

7.5 t ha-1

VC 4.2±2.2 4.7±1.8 3.4±0.9 4.6±2.3 4.1±1.3 3.9±0.7 2.8±1.5 2.9±1.4 2.4±0.6

IF 6.6±0.9 5.0±2.0 2.2±1.0 6.2±0.9 4.8±2.1 1.7±0.5 5.7±1.6 4.7±2.5 2.6±0.7

10 t ha-1

VC 8.4±4.5 8.7±5.7 6.0±1.7 9.2±3.3 7.2±3.3 5.6±1.7 4.7±1.8 4.3±1.7 2.7±0.6

IF 10.7±2.9 9.7±4.3 2.9±1.0 10.1±3.0 7.5±4.2 2.8±0.9 7.6±1.6 6.9±2.9 3.6±0.4

Control

0.4±0.1 0.9±0.4 1.0±0.6 0.4±0.1 0.9±0.4 1.0±0.6 0.4±0.1 0.9±0.4 1.0±0.6

F value

15.45***

9.304***

27.08***

24.59***

10.25***

29.18***

23.57***

11.71***

31.49***

*p<0.05,

**p<0.01,

***p<0.001,


146

Table 11. Changes in the ammonium (mg kg−1



Treatments

Lantana Ipomoea Paper waste


Layer

0-15 cm VC 24.5±11.2 27.2±18.2 7.1±6.1 33.8±19.2 30.3±19.7 6.5±4.5 12.3±5.1 12.1±7.1 3.2±2.6

IF 22.9±10.8 17.4±9.0 4.7±3.1 25.3±12.0 18.1±10.6 3.7±2.9 17.3±7.7 16.3±9.7 4.9±3.9

Cont. 2.2±0.9 0.6±0.2 0.6±0.2 2.2±0.9 0.6±0.2 0.6±0.2 2.2±0.9 0.6±0.2 0.6±0.2

15-30 cm VC 15.3±9.2 13.9±8.6 3.8±3.8 19.5±8.2 16.7±13.4 2.2±1.6 6.8±3.8 7.0±4.7 1.2±1.0

IF 12.7±5.2 8.7±6.1 1.4±0.9 15.0±6.7 10.4±7.0 1.7±1.6 9.3±4.2 8.4±5.5 1.6±1.4

Cont. 0.6±0.1 0.3±0.1 0.2±0.1 0.6±0.1 0.3±0.1 0.2±0.1 0.6±0.1 0.3±0.1 0.2±0.1

F value 8.024***

8.939***

7.134***

7.886***

7.671***

10.07***

10.42***

8.199***

8.217***

Amount

5 t ha-1

VC 10.0±5.3 9.9±6.8 2.1±1.4 19.1±8.2 16.3±9.4 3.1±2.5 6.2±3.7 6.9±4.8 0.9±0.8

IF 13.4±7.6 11.4±8.0 3.0±3.5 15.2±8.0 11.7±6.1 2.1±1.8 10.0±4.5 7.4±5.5 1.5±1.2

7.5 t ha-1

VC 21.3±9.0 21.1±13.1 4.3±3.4 22.5±14.7 20.6±16.3 3.5±3.4 8.8±4.3 8.7±5.6 2.3±1.8

IF 16.4±7.5 11.6±6.7 3.2±2.6 17.8±8.9 11.6±5.8 2.3±2.4 12.5±6.6 13.5±7.8 4.2±4.0

10 t ha-1

VC 28.4±9.9 30.6±18.1 10.0±6.2 38.5±18.6 33.6±22.5 6.4±5.2 13.7±5.1 13.0±7.6 3.4±2.9

IF 23.5±12.0 16.2±11.1 2.9±2.6 27.5±12.4 19.5±13.7 3.8±2.9 17.4±9.0 16.1±10.5 4.1±3.5

Control

1.4±1.0 0.4±0.2 0.4±0.3 1.4±1.0 0.4±0.2 0.4±0.3 1.4±1.0 0.4±0.2 0.4±0.3

F value

7.393***

6.507***

8.131***

5.877***

5.202***

3.792**

5.648***

4.928**

3.959***

*p<0.05,

**p<0.01,

***p<0.001,


147

Table 12. Changes in the nitrate (mg kg−1





Layer

0-15 cm VC 121.5±13.4 116.2±19.3 76.5±15.7 127.0±15.4 99.9±18.1 68.4±17.7 96.1±13.0 81.4±8.1 58.9±11.8

IF 116.8±17.2 112.3±30.9 73.5±15.6 114.7±18.0 110.6±31.1 67.3±13.6 98.8±12.5 99.1±16.4 66.8±12.1

Cont. 43.3±4.7 32.3±2.4 23.3±2.6 43.3±4.7 32.3±2.4 23.3±2.6 43.3±4.7 32.3±2.4 23.3±2.6

15-30 cm VC 106.0±12.7 103.0±15.3 70.0±11.9 108.3±7.5 97.7±13.5 63.3±14.9 86.3±7.9 77.4±9.3 57.0±12.1

IF 103.2±16.2 95.2±15.8 66.3±9.5 100.4±17.3 94.4±18.9 62.2±11.8 88.5±7.9 86.8±9.0 62.5±12.6

Cont. 39.7±5.4 28.2±1.9 23.1±3.6 39.7±5.4 28.2±1.9 23.1±3.6 39.7±5.4 28.2±1.9 23.1±3.6

F value 34.78***

27.63***

33.45***

36.50***

22.52***

20.91***

35.51***

56.18***

25.79***

Amount

5 t ha-1

VC 103.5±11.1 98.3±9.1 66.4±8.7 108.0±10.0 84.4±7.7 53.9±11.0 84.6±5.2 71.5±7.7 54.3±10.7

IF 97.9±11.9 86.4±5.7 63.9±8.1 95.5±11.9 85.8±7.8 60.6±9.3 86.9±8.1 85.6±10.7 61.1±11.0

7.5 t ha-1

VC 113.4±7.3 104.0±10.8 72.7±11.6 116.6±9.6 97.7±13.5 66.8±12.6 90.5±9.8 79.6±6.0 55.6±11.8

IF 107.1±15.7 97.9±15.4 65.9±8.5 99.6±13.5 93.3±13.4 62.0±10.0 91.1±10.2 91.3±12.7 63.4±11.5

10 t ha-1

VC 124.4±18.0 126.5±19.7 80.7±17.8 128.4±18.4 114.4±6.8 76.9±16.8 98.6±14.7 87.0±4.6 64.1±11.5

IF 125.0±14.8 127.0±29.6 79.9±15.9 127.6±11.1 128.4±29.7 71.6±16.1 102.9±10.6 102.1±15.5 69.4±14.0

Control

41.5±4.9 30.3±2.9 23.2±3.0 41.5±4.9 30.3±2.9 23.2±3.0 41.5±4.9 30.3±2.9 23.2±3.0

F value

30.11***

34.13***

28.78***

37.02***

37.76***

21.36***

26.77***

46.71***

19.38***

*p<0.05,

**p<0.01,

***p<0.001,


148

Table 13. Changes in the exchangeable phosphorus (mg kg−1

) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste,

or equivalent inorganic fertilizers (IF), at different periods of time (mean ± SD).



Layer

0-15 cm VC 4.35±0.31 4.00±0.26 3.64±0.25 4.11±0.22 4.41±0.67 3.60±0.54 4.08±0.18 4.04±0.29 3.75±0.29

IF 4.07±0.43 4.04±0.41 3.58±0.37 4.38±0.48 4.17±0.52 3.54±0.31 4.10±0.08 3.85±0.35 3.57±0.39

Cont. 3.98±0.38 3.81±0.24 3.61±0.14 3.98±0.38 3.81±0.24 3.61±0.14 3.98±0.38 3.81±0.24 3.61±0.14

15-30 cm VC 4.30±0.47 3.89±0.38 3.43±0.29 4.43±0.41 4.21±0.36 3.66±0.31 4.14±0.46 3.72±0.45 3.50±0.33

IF 4.27±0.19 3.77±0.74 3.39±0.43 4.40±0.69 3.86±0.44 3.48±0.27 4.14±0.29 3.97±0.23 3.76±0.26

Cont. 4.11±0.47 3.40±0.45 2.99±0.26 4.11±0.47 3.40±0.45 2.99±0.26 4.11±0.47 3.40±0.45 2.99±0.26

F value 17.72***

5.795**

0.864n.s

16.01***

5.341**

1.124n.s

16.56***

16.45***

1.471n.s

Amount

5 t ha-1

VC 4.34±0.40 3.74±0.36 3.52±0.33 4.29±0.41 4.24±0.59 3.46±0.39 3.92±0.40 3.93±0.50 3.56±0.26

IF 3.89±0.38 3.36±0.55 3.13±0.39 3.90±0.56 3.84±0.57 3.50±0.27 4.03±0.17 3.82±0.43 3.40±0.31

7.5 t ha-1

VC 4.26±0.28 4.05±0.32 3.51±0.29 4.25±0.40 4.40±0.51 3.64±0.36 4.38±0.28 3.79±0.38 3.54±0.44

IF 4.28±0.28 4.22±0.44 3.63±0.28 4.54±0.48 4.04±0.58 3.63±0.28 4.13±0.30 3.94±0.23 3.69±0.32

10 t ha-1

VC 4.38±0.51 4.05±0.19 3.59±0.26 4.29±0.33 4.29±0.56 3.78±0.52 4.04±0.16 3.92±0.36 3.79±0.22

IF 4.34±0.16 4.14±0.41 3.70±0.29 4.74±0.35 4.15±0.33 3.40±0.31 4.21±0.12 3.97±0.17 3.89±0.20

Control

4.04±0.39 3.60±0.40 3.30±0.38 4.04±0.39 3.60±0.40 3.30±0.38 4.04±0.39 3.60±0.40 3.30±0.38

F value

17.49***

6.239***

0.982n.s

24.98***

3.371**

1.374n.s

21.01***

11.75***

0.998n.s

*p<0.05,

**p<0.01,

***p<0.001,


149

Table 14. Changes in the exchangeable potassium (g kg−1

) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or




Layer

0-15 cm VC 0.42±0.02 0.38±0.02 0.36±0.01 0.41±0.03 0.39±0.02 0.36±0.02 0.41±0.02 0.38±0.02 0.36±0.02

IF 0.40±0.02 0.36±0.02 0.34±0.02 0.41±0.02 0.38±0.02 0.35±0.02 0.40±0.03 0.38±0.01 0.36±0.01

Cont. 0.40±0.01 0.36±0.02 0.34±0.01 0.40±0.01 0.36±0.02 0.34±0.01 0.40±0.01 0.36±0.02 0.34±0.01

15-30 cm VC 0.41±0.02 0.37±0.01 0.35±0.01 0.40±0.02 0.38±0.02 0.35±0.02 0.40±0.01 0.36±0.03 0.34±0.02

IF 0.42±0.02 0.37±0.02 0.33±0.03 0.40±0.03 0.39±0.02 0.36±0.01 0.40±0.02 0.38±0.01 0.35±0.01

Cont. 0.39±0.02 0.35±0.01 0.33±0.01 0.39±0.02 0.35±0.01 0.33±0.01 0.39±0.02 0.35±0.01 0.33±0.01

F value 2.846* 15.73

*** 19.93

*** 2.473

n.s 15.54

*** 18.62

*** 1.799

n.s 18.86

*** 28.68

***

Amount

5 t ha-1

VC 0.40±0.02 0.37±0.01 0.35±0.01 0.40±0.02 0.38±0.03 0.34±0.02 0.39±0.02 0.35±0.03 0.32±0.01

IF 0.39±0.02 0.35±0.01 0.31±0.01 0.39±0.02 0.38±0.01 0.35±0.02 0.39±0.01 0.38±0.01 0.36±0.01

7.5 t ha-1

VC 0.42±0.02 0.38±0.02 0.35±0.02 0.41±0.03 0.40±0.02 0.36±0.02 0.40±0.01 0.39±0.01 0.37±0.01

IF 0.41±0.02 0.36±0.02 0.33±0.01 0.41±0.01 0.38±0.01 0.35±0.02 0.40±0.02 0.38±0.01 0.35±0.01

10 t ha-1

VC 0.43±0.02 0.38±0.01 0.36±0.02 0.41±0.02 0.39±0.02 0.37±0.02 0.41±0.02 0.38±0.02 0.36±0.02

IF 0.42±0.02 0.38±0.01 0.35±0.02 0.42±0.02 0.39±0.03 0.36±0.02 0.41±0.02 0.38±0.01 0.35±0.01

Control

0.39±0.01 0.36±0.02 0.33±0.01 0.39±0.01 0.36±0.02 0.33±0.01 0.39±0.01 0.36±0.02 0.33±0.01

F value

2.142***

11.66***

26.79***

1.020n.s

9.598***

11.91***

1.015n.s

6.576***

17.52***

*p<0.05,

**p<0.01,

***p<0.001,


150

Table 15. Changes in the exchangeable calcium (g kg−1

) of potting soil amended with vermicast (VC) from lantana, ipomoea and paper waste, or




Layer

0-15 cm VC 8.20±0.82 7.11±1.00 5.73±1.02 7.78±1.17 6.66±1.69 5.58±0.91 7.91±0.72 7.15±1.08 6.40±0.95

IF 8.04±0.54 6.64±0.61 5.60±0.89 6.93±1.03 6.62±1.08 5.67±0.87 7.97±0.57 6.55±0.60 5.38±0.52

Cont. 6.32±1.69 5.15±0.45 4.81±1.14 6.32±1.69 5.15±0.45 4.81±1.14 6.32±1.69 5.15±0.45 4.81±1.14

15-30 cm VC 7.79±0.84 5.88±0.82 5.07±0.77 7.05±1.03 5.66±1.09 5.66±0.87 7.18±1.14 5.44±0.74 4.19±0.61

IF 7.61±0.87 6.21±0.97 5.31±0.99 7.72±0.63 6.45±1.42 5.94±1.12 7.92±0.60 5.78±0.76 4.74±0.62

Cont. 5.81±1.58 5.29±0.74 4.55±0.88 5.81±1.58 5.29±0.74 4.55±0.88 5.81±1.58 5.29±0.74 4.55±0.88

F value 2.409n.s

9.511***

24.44***

5.464**

2.488n.s

8.956***

12.49***

17.74***

17.90***

Amount

5 t ha-1

VC 7.41±0.84 5.97±1.05 5.13±0.90 6.81±1.54 5.60±1.36 5.26±0.91 6.96±1.20 5.99±0.96 4.60±1.13

IF 7.37±0.63 5.97±0.97 4.73±0.69 6.62±1.07 5.82±0.81 5.67±0.92 7.81±0.53 5.87±0.87 4.91±0.52

7.5 t ha-1

VC 7.97±0.47 6.69±1.10 5.64±1.19 7.64±1.05 6.18±1.36 5.92±1.03 7.52±0.84 6.53±1.25 5.84±1.27

IF 7.89±0.88 6.35±0.71 5.23±0.39 7.34±0.63 6.93±1.30 5.94±0.94 7.66±0.68 6.19±0.71 5.13±0.78

10 t ha-1

VC 8.60±0.75 6.83±1.06 5.43±0.75 7.77±0.49 6.69±1.67 5.67±0.60 8.16±0.61 6.36±1.60 5.44±1.52

IF 8.21±0.49 6.95±0.47 6.40±0.76 8.03±0.42 6.86±1.34 5.80±1.18 8.37±0.13 6.44±0.73 5.14±0.66

Control

6.06±1.49 5.22±0.57 4.68±0.97 6.06±1.49 5.22±0.57 4.68±0.97 6.06±1.49 5.22±0.57 4.68±0.97

F value

8.544***

11.47***

13.7`***

3.884**

0.908n.s

4.822***

31.33***

19.17***

14.93***

*p<0.05,

**p<0.01,

***p<0.001,


151

Table 16. Changes in the micronutrient content (mg kg−1

) of potting soil amended with vermicast (VC) from lantana or equivalent inorganic

fertilizers (IF), at different periods of time (mean ± SD).

Treatments Mg Cu Fe Mn Zn B

Initial Final Initial Final Initial Final Initial Final Initial Final Initial Final

Layer

0-15 cm VC 99.8 63.1 6.55 2.76 59.3 52.8 65.0 36.8 58.7 26.7 28.3 12.4

IF 98.4 49.0 6.10 1.48 58.3 46.7 60.3 33.0 57.4 24.9 28.1 10.3

Cont. 88.0 43.4 5.09 1.24 55.4 49.4 46.2 37.4 53.5 25.5 26.0 13.8

15-30 cm VC 91.9 48.6 5.25 2.10 56.9 44.2 52.2 29.4 56.0 24.3 25.8 11.2

IF 88.2 56.3 5.52 1.80 55.4 40.5 54.3 30.9 56.3 24.1 26.2 10.5

Cont. 86.3 40.9 5.04 3.45 58.3 54.6 46.0 28.2 54.8 22.2 27.2 14.9

F value 5.833**

7.054**

18.47***

34.16***

2.243n.s

2.497n.s

13.78***

7.194**

8.757**

3.569* 2.668

n.s 1.447

n.s

Amount

5 t ha-1

VC 91.3 50.5 5.67 2.30 56.4 40.5 54.2 30.7 56.6 24.9 26.7 12.4

IF 88.9 51.5 5.58 1.24 55.2 44.7 53.8 29.8 55.7 23.9 26.2 13.0

7.5 t ha-1

VC 93.8 53.0 5.79 2.50 57.5 46.7 58.1 36.5 57.4 25.7 26.7 11.1

IF 93.6 53.5 5.83 1.77 56.7 40.6 57.3 31.3 57.2 24.7 27.8 9.5

10 t ha-1

VC 102.4 64.1 6.23 2.50 60.2 58.3 63.4 32.2 58.1 25.9 27.7 11.9

IF 97.4 53.0 6.02 1.91 58.5 45.4 60.8 34.8 57.8 24.9 27.4 8.7

Control

87.2 42.2 5.06 2.35 56.9 52.0 46.1 32.8 54.1 23.9 26.6 14.4

F value 3.413* 2.788

n.s 1.582

n.s 0.640

n.s 1.914

n.s 3.818

* 4.430

* 0.430

n.s 3.627

* 0.672

n.s 0.364

n.s 3.404

n.s

*p<0.05,

**p<0.01,

***p<0.001,


152


) of potting soil amended with vermicast (VC) from ipomoea or equivalent inorganic


Treatments Mg Cu Fe Mn Zn B


Layer

0-15 cm VC 103.9 50.7 5.42 2.67 72.1 41.4 55.8 25.6 56.9 25.6 31.0 15.2

IF 98.0 44.4 5.35 1.75 69.4 49.7 53.1 31.0 55.6 27.4 31.3 14.6

Cont. 88.0 43.4 5.09 1.24 55.4 49.4 46.2 37.4 53.5 25.5 26.0 13.8

15-30 cm VC 90.3 40.6 5.08 2.78 56.7 32.0 50.1 21.5 55.0 23.6 26.4 11.9

IF 88.6 40.7 5.07 2.30 57.0 49.0 48.8 26.3 54.5 24.1 27.0 14.4

Cont. 86.3 40.9 5.04 3.45 58.3 54.6 46.0 28.2 54.8 22.2 27.2 14.9

F value 3.600* 5.958

** 2.129

n.s 18.86

*** 20.27

*** 6.841

** 6.324

** 33.98

*** 3.544

* 3.071

n.s 4.668

* 1.110

n.s

Amount

5 t ha-1

VC 86.3 46.4 5.17 2.32 61.7 32.0 49.8 25.1 54.5 22.9 27.2 14.2

IF 89.0 40.0 5.41 2.12 60.9 52.8 48.0 27.8 53.8 26.8 27.1 15.4

7.5 t ha-1

VC 99.0 44.1 5.24 2.98 63.7 41.2 52.8 21.4 56.6 24.8 28.3 14.5

IF 92.4 44.2 5.10 2.13 64.2 52.0 50.3 29.9 55.1 25.4 29.2 14.6

10 t ha-1

VC 106.0 46.6 5.34 2.88 67.7 36.8 56.3 24.2 56.6 26.1 30.6 11.9

IF 98.7 43.4 5.11 1.81 64.6 43.3 54.5 28.1 56.1 25.0 31.1 13.5

Control

87.2 42.2 5.06 2.35 56.9 52.0 46.1 32.8 54.1 23.9 26.6 14.4

F value 3.552* 0.514

n.s 0.722

n.s 0.503

n.s 1.065

n.s 5.443

** 5.502

** 1.846

n.s 2.186

n.s 1.003

n.s 1.186

n.s 0.605

n.s

*p<0.05,

**p<0.01,

***p<0.001,


153


) of potting soil amended with vermicast (VC) from paper waste or equivalent inorganic


Treatments

Mg Cu Fe Mn Zn B


Layer

0-15 cm VC 105.7 57.4 5.47 2.52 65.8 35.8 52.5 36.2 56.5 22.2 26.4 6.2

IF 105.2 58.3 5.18 2.67 63.6 33.2 55.4 31.9 55.8 23.7 26.5 7.4

Cont. 88.0 43.4 5.09 1.24 55.4 49.4 46.2 37.4 53.5 25.5 26.0 13.8

15-30 cm VC 90.2 54.1 5.09 2.04 59.1 31.0 47.8 28.6 54.9 21.5 25.3 6.2

IF 87.6 43.4 5.15 1.37 58.1 28.0 49.5 29.8 55.0 21.7 26.3 11.7

Cont. 86.3 40.9 5.04 3.45 58.3 54.6 46.0 28.2 54.8 22.2 27.2 14.9

F value 4.861* 2.601

n.s 3.406

* 35.22

*** 8.143

** 68.54

*** 10.49

*** 7.154

** 3.785

* 2.894

n.s 0.875

n.s 3.664

*

Amount

5 t ha-1

VC 93.2 51.3 5.17 2.03 60.0 33.2 48.3 29.6 54.4 21.7 25.7 7.9

IF 90.0 45.5 5.07 1.77 57.9 29.6 50.1 28.2 54.8 22.1 26.7 10.3

7.5 t ha-1

VC 93.4 56.5 5.42 2.24 62.1 32.9 50.7 31.4 56.3 22.2 25.2 5.9

IF 94.8 51.9 5.11 2.04 61.5 28.8 53.3 30.7 55.0 22.7 26.0 11.1

10 t ha-1

VC 107.1 59.4 5.25 2.56 65.2 34.1 51.6 36.1 56.4 21.5 26.7 4.8

IF 104.3 55.3 5.31 2.24 63.1 33.4 54.0 33.7 56.2 23.4 26.5 7.2

Control

87.2 42.2 5.06 2.35 56.9 52.0 46.1 32.8 54.1 23.9 26.6 14.4

F value 1.746n.s

1.275n.s

1.352n.s

0.151n.s

3.043n.s

17.23***

3.248* 0.540

n.s 2.065

n.s 0.797

n.s 0.920

n.s 2.721

n.s

*p<0.05,

**p<0.01,

***p<0.001,


154

treatments; whereas, in the 15–30 cm layer a higher

Caext content was recorded with IF treatments. The

concentration of trace nutrients such extractable

form of magnesium (Mgext), copper (Cuext), iron

(Feext), manganese (Mnext), zinc (Znext) and boron

(Bext) significantly increased in soil with both VC

and IF fertilization (Tables 16-18). The Mgext

content in soil increased to the extent of 19% with

paper waste based VC and its equivalent IF

treatments. Increase in Mgext with ipomoea and

lantana based treatments were up to 18 and 15%,

respectively. At the end of the experiment, the

Mgext with ipomoea based treatments showed

higher reduction of 56% followed by 50 and 46%

with paper waste and lantana based treatments.

Fertilization with VC and IF, increased the Cuext

content to the extent of 70%. Finally, a maximum

reduction of about 19% in Cuext content was

recorded in lantana based treatments; whereas, the

paper waste and ipomoea based treatments showed

only 7% maximally.

Lantana, paper waste and ipomoea based VC

and its equivalent IF treatments increased the Feext

content of soil to the extent of 23, 16 and 7%,

respectively. In the case of Znext up to 7, 4.4 and

4.1% increase was recorded with lantana, ipomoea

and paper waste based treatments. Except the

ipomoea based treatments, in all others, the Feext

and Znext depletion was lower in the VC compared

to corresponding IF treated soils. Similarly, the

ipomoea and paper waste based VC treatments

showed lesser reduction in Mnext compared to its

equivalent IF treatments; whereas, the lantana

based VC showed higher reduction. By the end of

the experiment, the reduction in Mnext was to the

extent of 60, 59 and 43% with ipomoea, lantana and

paper waste based treatments, respectively. The

increase of about 14.5% in Bext content was in

lantana based treatments, and those of ipomoea and

paper waste showed 3 and 0.4%, respectively. The

higher Bext content in lantana based treatments

could be the reason for better growth of

experimental plants with maximum nodulation.

Increase in availability of Bext was reported to

enhance N fixation in the nodules of soybeans

(Yamagishi and Yamamoto, 1994). Reduction in

Bext content after plant harvest was 82, 68 and 61%

with paper waste, lantana and ipomoea based

treatments, respectively. The trace nutrients, such

as Cuext, Feext, Znext and Bext depleted maximally in

IF based treatments, in both surface and deeper soil

layer; whereas Mgext and Mnext content of surface

soil layers reduced maximally with IF treatments

and in the deeper soil layer VC showed higher

reduction. In all the cases, the increasing dose of

nutrient application exaggerated its changes as

stated above, and there was no differential trend of

results with different dose of VC and IF

application.

The nutrient status of soil is influenced by

several factors. Amongst the nutrient up take by

plants, nutrient leaching from soil, microbial

immobilization and mineralization are considered

to influence the fate of nutrients in the soil largely

(Carlile and Wilson, 1993). In these experiments,

lower concentration of many nutrients in IF treated

soils may be due to high leaching of mineral

nutrients from the soil. Organic matter inputs

through VC, in addition to supplying nutrients,

improve soil aggregation, and stimulate microbial

diversity and activity (Shiralipour et al., 1992;

Carpenter-Boggs et al., 2000). The changes in the

physical and microbial properties of soil influence

many chemical and biological reactions (Sharma

and Bhushan, 2001). In the present study, increase

in microbial and enzymatic activity with

vermicompost application (Masciandaro et al.,

1997; Arancon et al., 2006) might have mineralized

the organic bound nutrients, which attributed to

increase in the mineral nutrient content in soil

together with improved growth and productivity of

experimental plants.

4. Conclusions

The paper describes the impact of

vermicompost derived from different parent

materials on the physical and chemical qualities of

155

soil which are directly related to the plant growth.

The impact of the vermicompost on soil health in

terms of its physical and chemical qualities were

compared with that of an inorganic fertilizers which

had all the main macro- and micro-nutrients in

concentrations equivalent to the ones present in the

vermicompost. Several sets of experiments were

carried out in which the vermicast generated from

different organic wastes such as paper waste, leaves

of ipomoea (Ipomoea carnea), and of lantana

(Lantana camara) or inorganic fertilizers were

applied in potting soil housing cluster bean

(Cyamopsis tetragonoloba). Samples from all these

treatments were collected on weekly basis during

different stages of plant growth. The results reveal

that vermicast application created a suitable

physical environment by reduction in bulk density

and improving the water holding capacity and

porosity of soil used. In addition, throughout the

experiment, the nutrient content of soil was

significantly higher compared to inorganic

fertilizers treated one. Although, the inorganic

fertilizers application initially increased the nutrient

content in soil, as the days progressed, substantial

quantity of applied nutrient became unavailable to

the plants, probably it lost due to high leaching of

mineral nutrients from the soil. Apparently, no

significant impact has been observed on physical

properties of soil with inorganic fertilizers

application. Consequently, the vermicompost

amendment may be considered a good strategy for

improving plant growth which reduces the

deterioration of agricultural lands due to rampant

use of inorganic fertilizers.

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EFFECT OF STORAGE ON SOME PHYSICAL AND

CHEMICAL CHARACTERISTICS OF VERMICAST: A

PRELIMINARY STUDY

Chapter

10

159

A paper based on this chapter has been accepted for publication in

Journal of Applied Horticulture

CChhaapptteerr 1100

Effect of storage on some physical and chemical

characteristics of vermicast: A preliminary study

Abstract

It is widely acknowledged that vermicast has beneficial effect on plant growth but little is known on how the

manner and duration of storage affect the vermicast quality. Investigating the impact of storage on the

characteristics of castings is of great importance with respect to understanding its optimum age for best

utilization. In an attempt to cover this knowledge-gap we have carried out studies on the morphology, water-

retention capacity, and availability of vermicast nutrients as function of vermicast ageing when the latter is

stored. The study reveals that during storage of vermicast, the physical and chemical properties of castings

get altered. The prolonged storage period reduced the nutrient concentration and in turn the beneficial

properties of vermicast required for plant growth.

1. Introduction

Vermicomposting is the term given to the

process of conversion of biodegradable matter by

earthworms into vermicast (Abbasi and Ramasamy,

2001). In the process, a major fraction of the

nutrients contained in the organic matter is

converted to more bioavailable forms. Application

of vermicompost improves the soil structure by

increasing porosity and reducing the bulk density. It

improvises soil aeration, water-holding capacity,

buffer capacity, and cation exchange capacity of

soil (Nada et al., 2011). In addition, the

vermicompost is also reported to contain

biologically active substances such as plant growth

regulators and have been shown to increase growth

of many plants (Tomati et al., 1983, 1988, 1990;

Tomati and Galli, 1995; Abbasi and Ramasamy,

1999; Atiyeh et al., 2002; Arancon et al., 2003,

2004; Gajalakshmi and Abbasi, 2004; Acevedo and

Pire, 2004; Edwards, 2004; Sinha, 2009).

Although a considerable number of studies

have been carried out on vermicomposting of

various organic materials with different earthworm

species and their impact on the soil and plant

growth (Logsdon, 1994; Sundaravadivel and Ismail,

1995; Gajalakshmi et al., 2001a,b, 2002; Singh and

Sharma, 2002; Gajalakshmi and Abbasi, 2003,

2004; Suthar, 2006, 2007; Padmavathiamma et al.,

2008), there is still a lack of knowledge on the

change in the properties of vermicast during the

course of storage. The castings of anecic and

endogeic earthworms have been extensively studied

in relation to the changes in the physico-chemical

properties during ageing process (Hindell et al.,

1997; Decaëns et al., 1999; Decaëns, 2000; Aira et

al., 2005; Mariani et al., 2007). There are also few

studies on the enzymes and microbial aspects of

castings generated from the epigeic earthworms

(Parthasarathi and Ranganathan, 1998, 1999;

Scullion et al., 2003). There is no study on changes

in the characteristics of vermicast during the ageing

as a function of manner of storage. Investigating

this storage effect on castings is of great importance

with respect to understanding the changes in

nutrient status and physical properties of vermicast.

The present work has been taken up to investigate

160

the changes in the properties of the vermicast when

stored so that the optimum age for best utilization

could be understood. The present study was

conducted with vermicast generated from neem

leaves.



Pondicherry University campus and its vicinity.

The collected leaves were washed with water to

remove adhering material and soaked for 48 hours

in order to remove phenolic compounds and to

make substrate softer and palatable to earthworms

(Agarwal et al., 1987; Nath et al., 1987; Agarwal et

al., 1991). Rectangular wooden boxes (depth 30

cm, width 35 cm, length 39 cm) were used as

vermireactors. The reactors were filled from bottom

up with successive layers of coarse sand and soil to

a thickness of 3 and 5 cm, respectively. Neem

leaves was added as feed with an epigeic

earthworm species Eudrilus eugeniae. After 2

weeks, the vermicast was harvested. An aliquot of

fresh castings was analyzed immediately whilst the

rest were stored for <7, 14, 21, 60, 90 and >120

days. The casts were stored in the polyethylene

bags of 20 micron thick and 25 x 18 cm size.

Plastic bags filled with 500 g of vermicast were

stored at room temperature in order to imitate the

general way of storage of vermicomposting in

commercial sectors.

Moisture content of castings was determined

by weight loss at 105°C. To estimate bulk density

(pBulk), sample volume was measured with a

graduated cylinder and its dry weight determined

by oven drying. The particle density (pParticle) was

determined by volumetric flask method (Bashour

and Sayegh, 2007). The quotient value of weight of

the sample and its volume which was measured

through volume of water displaced by known

amount of soil sample in the volumetric flask is

reported as particle density. To measure the water

holding capacity (WHC), the samples were filled in

cylinders with a perforated base and immersed in

water and drained. The quantity of water taken up

by samples is determined by drying to constant

mass at 105°C (Margesin and Schinner, 2005). The

total and water filled porosity were calculated from

the particle and bulk density values of the

respective samples, using the following equation

(Carter and Gregorich, 2008).

….…….…….… (1)

………. (2)

.(3)

were, Db is the bulk density, Dp is the particle

density, θw is gravimetric water content, and Dw is

the density of water at corresponding temperature.

Electrical conductivity (EC) and pH were

measured with suspensions of samples in water

(1:2, w/v) (Bashour and Sayegh, 2007) by using

EI™ 611E EC meter and Digison™ digital pH

meter 7007, respectively. Thin sections of casts

were made after impregnating the samples in

araldite using Bueller PetroThin™ thin sectioning

system (FitzPatrick, 1993). The internal and

external structures of thin sectioned castings were

observed under the binocular microscope.

Total organic carbon (Corg) was determined

following modified dichromate redox method

(Heanes, 1984). External heating was applied

during the oxidation process in order to quicken

and complete oxidation of organic carbon in the

sample. Total nitrogen (Ntot) was determined by

modified Kjeldahl method (Kandeler, 1993) using

Kel Plus™ semi-automated digester and distillation

units. In order to include nitrate, nitrite, nitro and

nitroso groups in the assay, a mixture of salicylic

acid and sulphuric acid was used for digestion. All

the elemental present in the vermicast were

analyzed by Bruker™ S4-Pioneer model

wavelength dispersive X-ray fluorescence

spectrophotometer (WD-XRF). The samples were

ground to particle size well below 100 µm using

ball mill in order to minimize the grain size

interference on XRF-measurement. The

161

concentration of major elements found in the

vermicast is reported in this chapter. The data were

analyzed statistically with software SPSS 16

package and subjected to one-way ANOVA.

Comparisons between means were tested with LSD

test.


Fresh castings of E.eugeniae produced from

neem leaf litter were fine, long, slender and pellet-

like in structure. The size was in the range of 0.1 to

2.0 mm length and 0.1 to 1.2 mm breadth. During

storage, the casts had undergone significant

changes in their physical and chemical properties

(p< 0.001) (Tables 1-2). The castings stored for

prolonged period (>120 days), had its structure

disintegrated and formed clod like aggregates. The

initial moisture content of 77.4 ± 3.5% was reduced

to 66.7±2.5% in 60 days stored vermicast.

Afterwards, the moisture content drastically

decreased to about 20% in the 90 and >120 days of

storage. Decrease in the moisture content of

vermicast during the storage may exert a strong

influence on the microorganisms (Nannipieri et al.,

2003) and their enzyme activity in the vermicast

(Parthasarathi and Ranganathan, 1998). The

microbiota and their enzymes in turn reflect on the

mineralization of nutrient (Birch, 1964). The results

indicate that the drastic loss in moisture content

(>75%) after 90 days leads to reduction in

microbial mediated activity in the vermicast.

In the first 3 weeks of storage, the particle

density (pParticle) was stable, after that it increased

and became twofold high at > 120 days stored

vermicast compared to the fresh castings (p<0.001)

(Table 1). Ruhlmann et al. (2006) has reported that

the pParticle of castings varied considerably due to

the degree of decomposition of organic matter

present in it. In the present study the pParticle was

ranging from 1.359 to 2 g cm-3

, which is very lower

than the soil pParticle range of 2.6–2.8 g cm-3

in

relation to the plant growth. Low pParticle of the

vermicast can be explained by their high Corg

content. It has been reported that increase of Corg in

the soil, decrease the pParticle at the rate of 0.04-0.06

g cm-3

per percentage of Corg (Ruhlmann et al.,

2006).

As the age of the vermicast progressed, there

was a significant increase in bulk density (pBulk)

(p<0.001; Table 1). During the first week of

storage, the pBulk was 0.307 g cm-3

, and then it

increased, till the end of the experiment

(0.603±0.014 g cm-3

). There was twofold increase

in pBulk in four months of storage. However, it is

lower than the soil pBulk range of 0.7–1.8 g cm-3

in

relation to the plant growth (Lal and Shukla, 2004).

Low pBulk is desirable for plant growth, as it makes

easier for plant root penetration and it posses high

water infiltration rates. On the other hand, high pBulk

would impede root penetration and reduce the air

and water movement (Edwards, 2004).

Porosity of castings decreased distinctly

(p<0.001) throughout the >120 days of storage and

its range was between 69.9 to 77.4% (Table 1).

Porosity directly influences water infiltration,

hydraulic conductivity and water storage capacity

in soils (Blanchart et al., 2004). Moreover, it

greatly influences the structure, function and

interaction of microbial and microfaunal

communities (Hattori, 1994). The percentage of

water-filled pore space (WFPS) was high, but not

significantly different in the first 21 days of storage,

ranging between 30.5 and 33.4%. High WFPS may

decline the microbial activity, presumably as a

result of additional water presenting a barrier for

diffusion of oxygen and the waste products away

from microorganisms (Linn and Doran, 1984a,b;

Doran, 1990). The castings stored for 90 and >120

days showed 14.9 and 18.2% of WFPS,

respectively; this lower WFPS is also not suitable

for the microbial activity. Usually excessive

dryness is more prejudicial to microorganisms than

an excess of water-filled pores (Tate, 1985;

Paradelo and Barral, 2009).

WHC was the maximum in castings stored

162

Table 1. Physical characteristics (mean± SD) of castings stored for different periods and the calculated F-

values using one-way ANOVA

*p<0.05,

**p<0.01,

***p<0.001,


for 2 weeks and it was 651.0 ± 37.4 and 798.0 ±

9.0% in the first and second week, respectively

(Table 1). High WHC of the castings during the

initial period of storage may be due to the

abundance of micropores present in the castings

(Chaudhuri et al., 2009). At the end of the

experiment nearly 6 fold decrease in WHC was

recorded compared to fresh castings (p<0.001).

While estimating the WHC of the vermicast stored

for 90 and > 120 days, it was observed that the

vermicast did not absorb water for many hours,

indicating high degree of water repellency. This

may be due to conformational rearrangements of

the organic matter (Mashum and Farmer, 1985;

Valat et al., 1991; Roy et al., 2000), and excess

dryness (King, 1981; Ritsema et al., 1998; De

Jonge et al., 1999; Bachmann and van der Ploeg,

2002; Quyum et al., 2002).

Electrical conductivity indicates the

concentration of total soluble salts in solution, thus

reflecting the degree of soil salinity and it affects

plants at all stages of development. The sensitivity

may vary from one growth stage to another for

some crops (Maas and Hoffman, 1977). In the

neem castings of our study, maximum EC of

4.893±0.210 mmhos cm-1

was recorded in the fresh

castings and it significantly declined during the

storage (p<0.001) (Table 1). There was a maximum

of about 68% reduction in EC during first 21 days

of storage and thereafter there was a slow and

steady decline till the end of the experiment.

Table 2. Total nitrogen and organic carbon (mean± SD) of castings stored for different periods and the

calculated F-values using one-way ANOVA

Parameter Days of storage

F value < 7 14 21 60 90 >120

Total Kjeldahl

nitrogen mg g-1

21.77 ± 1.33 14.95 ± 0.35 14.33 ± 1.46 12.55 ± 2.19 13.94 ± 2.49 11.20 ± 0.83 239.5

***

Total organic carbon

mg g-1

108.0 ± 2.1 115.2 ± 8.7 130.8 ± 2.5 144.6 ± 5.4 151.2 ± 1.7 90.0 ± 1.9 46.5

***

*p<0.05,

**p<0.01,

***p<0.001,


Days of

storage

Water Content

%

Bulk Density

g cm-3

Particle Density

g cm-3

Pore Space

%

Water-Filled

Pore Space

%

Water Holding

Capacity

%

EC

mmhos cm-1

<7 77.44 ± 3.46 0.307 ± 0.006 1.359 ± 0.034 77.41 ± 0.94 30.49 ± 1.77 651.0 ± 37.4 4.893 ± 0.210

14 76.67 ± 0.84 0.317 ± 0.009 1.349 ± 0.33 76.47 ± 0.70 31.63 ± 1.14 798.0 ± 9.0 3.673 ± 0.361

21 75.70 ± 3.86 0.334 ± 0.005 1.350 ± 0.012 75.26 ± 0.48 33.41 ± 2.06 592.1 ± 30.8 1.569 ± 0.023

60 66.73 ± 2.45 0.383 ± 0.014 1.435 ± 0.037 73.31 ± 0.86 34.65 ± 1.57 518.0 ± 20.6 1.526 ± 0.050

90 20.63 ± 3.49 0.524 ± 0.015 1.849 ± 0.028 71.68 ± 0.54 14.87 ± 2.94 132.7 ± 5.0 1.489 ± 0.029

>120 21.36 ± 2.82 0.603 ± 0.014 2.000 ± 0.037 69.86 ± 0.70 18.24 ± 2.94 117.8 ± 9.0 1.439 ± 0.040

F value 772.1***

1034.7***

774.6***

145.9***

135.5***

1441.1***

649.4***

163

Table 3. Major elements present in the casting stored for different periods (expressed in %).

Elements Days of storage

< 7 14 21 60 90 >120

Ca 60.18 60.14 60.09 48.91 17.444 15.56

Al 2.280 1.260 1.190 1.120 1.042 1.045

Cl 3.890 3.850 3.790 3.150 0.898 0.774

Fe 3.773 2.505 2.382 1.878 1.643 1.108

K 12.92 14.77 14.85 10.69 2.580 2.864

Mg 2.578 3.716 3.848 2.443 0.633 1.111

Mn 0.258 0.193 0.179 0.199 0.070 0.245

Na 0.429 0.428 0.428 0.397 0.394 0.436

P 2.090 1.460 1.430 1.430 1.425 1.230

S 5.360 4.169 4.146 4.058 1.060 1.480

Si 15.63 15.54 15.38 15.01 13.38 12.82

Ti 0.888 0.446 0.368 0.133 0.105 0.070

Zn 0.377 0.093 0.090 0.090 Nil Nil

The total nitrogen content in fresh castings

was 21.77±1.33 mg g-1

(Table 2). In the 14 days

stored castings, 31% reduction in Ntot was observed.

Afterwards the concentration decreased slightly

during the entire storage process. The gaseous loss

of nitrogen from the casts may be the reason for the

maximum Ntot loss during 14 days of storage. A

higher loss of N during the initial weeks was likely

due to the intense ammonia volatilization. In

general, NH3+ volatilization is strongly dependent

on the NH3+ and NH4

+ concentration (Pagans et al.,

2006). It has been reported that up to 30% nitrogen

loss occurs in fresh castings by denitrification

(Kharin and Kurakov, 2009). The organic carbon

content of fresh castings was 108.0±2.1 mg g-1

.

During the storage process, the Corg content in the

castings significantly increased up to 29% in the

course of 90 days. After 120 days, there was a

drastic decrease and its concentration was near to

those of fresh castings (Table 2).

Decaëns et al. (1999) observed same pattern

of Corg increase but without any decline over

prolonged ageing of vermicast of large species of

anecic earthworm, Martiodrilus carimaguensis.

They have also summarized a combination of

several factors for Corg increase during the ageing

process. Among those factors, the possible reasons

that would be applicable in the present studies are

fixation of atmospheric CO2 from autotrophic

microorganisms, such as algae or nitrification

microorganisms (Vinceslas- Akpa and Loquet,

1997) and accumulation of organic matter by cast-

dwelling macroinvertebrates. The significant

decrease of Corg in vermicast stored for more than

120 days can be attributed to excessive dryness of

castings, which is not suitable and beneficial to

microbiota. The fresh castings are noted to be

enriched in Ca, followed by K (Table 3). The

concentration of these elements was almost stable

until 21 days of storage. Afterwards, there was a

decline throughout the study. Lal and de

Vleeschauwer, (1981) and Schrader and Zhang

(1997) ascribed the high Ca content of casts to the

presence of calcite spheroids originating from

worms’ calciferous glands which regulates the CO2

in their tissues (Briones et al., 2008).

In the fresh castings, 12.9% of K was

recorded and there was 77% of loss at the end of 4

months storage. In the case of P, the initial

concentration of 2.1% was reduced to 1.2%, at the

end of 4 months. A maximum P loss of 73% was

observed in the first two weeks of storage. After

that, there was slow and steady decline of P content

till the end of the experiment. Except Mg, the

164

concentration of other elements such as Al, Fe, Na,

Cl, S, Si, Ti and Zn decreased as the storage period

increased (Table 3). Changes in Mg concentration

did not show any trend as the age of the castings

progressed. The overall results show that 60 days of

storage did not show much variation; after that

there was a significant decrease in the concentration

of these elements. The reduction of K, P and other

metals content may be due to nutrient assimilation

by the bacterial and fungal grazing macro-

invertebrates in the castings. Loss of these trace

nutrients may slide down the positive impact of

vermicompost on the plant growth.

4. Conclusions

The present study reveals that during storage

of vermicast, the physical and chemical properties

of castings get altered. The prolonged storage

period reduced the nutrient concentration and in

turn the beneficial properties required for plant

growth. According to the results of this study, most

of the characteristics of the castings are retained

during the first 60 days of storage. Further as

storage was continued, the nutrient status depleted.

The changes in physical properties are

disintegration of the structure of the vermicast,

increase in bulk density, water repellency, decrease

in water holding capacity and water content. All

these factors lead to adverse impact on plants when

applied as manure. Therefore, utilization of

vermicast before nutrient loss is recommended or

castings need to be stored by appropriate methods

which should prevent the loss of nutrient

concentration and maintain the physical

characteristics of vermicast. At present, there are no

prescribed guidelines for storage of castings; hence

comprehensive method of storage needs to be

explored extensively.

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EFFECT OF STORAGE ON THE PROPERTIES OF

VERMICOMPOST GENERATED FROM PAPER WASTE –

WITH FOCUS ON PRE-DRYING AND EXTENT OF

SEALING

Chapter

11

169


International Journal of Energy and Environmental Engineering, DOI: 10.1007/s40095-014-0135-z

CChhaapptteerr 1111

Effect of storage on the properties of vermicompost generated

from paper waste – with focus on pre-drying and extent of

sealing

Abstract

In chapter 10, a preliminary study on the impact of storage of vermicast of different age, without any

pretreatment was reported. The study revealed that the physical and chemical properties of vermicast get

altered during storage thereby reducing the fertilizer value of vermicast. In the present study the effect of pre-

drying and type of packing is reported. Vermicast generated from paper waste was packed in airtight and

partially sealed bags with and without pre-drying for 24 hours. Changes in several physical, chemical, and

biological properties of the castings were monitored for three months with weekly assessments. The results

reveal that the beneficial properties of vermicast were the highest when it was fresh. There was deterioration

on storage, which can be minimized if the castings are contained in airtight bags after pre-drying the casts.

1. Introduction

The vermicast that is deposited by the

earthworms on the soil is known to fertilize the soil

as well as influence its physical and chemical

properties in a way that is beneficial to plant growth

in particular and soil environment in general. Due

to this realization several studies have been

conducted on the fate of vermicast, especially how

the biological, chemical, and physical attributes of

the vermicast change with time (Hindell et al.,

1997a; Decaëns et al., 1999; Parthasarathi and

Ranganathan, 1999, 2000; Decaëns, 2000; Tiunov

and Scheu, 2000; Scullion et al., 2003; Aira et al.,

2005, 2010; Mariani et al., 2007; Kawaguchi et al.,

2011). These studies have been on either vermicast

generated from non-specific substrates in nature or

from blends of soil and phytomass. The focus of the

studies has been primarily on the stability of

vermicast generated by anecic and endogeic

(geophagous and geophytophagous) earthworm

species as such casts are rich in soil and influence

the stability of biogenic structures. Very few

studies have been done on epigeic or phytophagous

(‘humus feeder’) species. Moreover, when

vermicast is deposited in nature, its fate is strongly

influenced by (a) soil dwelling invertebrates –

which colonize the vermicast and feed upon the

organic matter it contains (Decaens et al., 1999;

Decaëns, 2000); (b) vegetation – which takes up the

nutrients from the castings (Jiménez and Decaëns,

2004); (c) soil microbes including autotrophic

algae, nitrification bacteria or fungi – which are

involved in fixation of atmospheric CO2 (Jiménez

and Decaëns, 2004); (d) immobilization or

mineralization of nutrients in the vermicast (Kharin

and Kurakov, 2009), and (e) environmental factors

such as rain, flooding, or drought.

A few controlled studies have been reported

on the change in the properties of vermicast upon

ageing (Shipitalo and Protz, 1989; Marinissen and

Dexter, 1990; Marinissen et al., 1996; Hindell et

al., 1997a,b; McInerney and Bolger, 2000; Tiunov

and Scheu, 2000). These studies have primarily

aimed to simulate the conditions which the biogenic

170

structures experience in nature. For this the casts

were generated from either soil or blends of soil

and phytomass and stored in soil/sand columns. It

was seen that the soil particles present in the casts

get chemically bound with organic matter, perhaps

through chelation, which increases the stability of

the casts. It also protects the organic matter content

of the casts from decomposition (Shipitalo and

Protz, 1989; McInerney et al., 2001), as the organic

matter that is attached to the minerals with strong

chemical bounds is less accessible to

microorganisms (Kaiser et al., 2007). In addition,

extracellular enzymes are protected from

degradation and proteolysis by the clay minerals

contributed by the soil (Nannipieri and Smalla,

2006).

In contrast to the focus of the prior art

summarized as above, the conditions associated

with the storage of vermicast when it is produced

by anthropogenically controlled vermicomposting

and for the specific purpose of use as a fertilizer are

very different. The concern here is to ensure that

the vermicast retains as many and as much of the

plant-friendly attributes as does fresh vermicast and

the physical integrity of the cast is not of much

significance. The only pre-existing studies on the

effect of storage on vermicompost (Parthasarathi

and Ranganathan, 1999, 2000), have been based on

the use of two-month old press-mud as feed for

earthworms, and assessment of the changes in

major nutrients (N,P and K), microbial activity and

enzyme activity of the vermicast that was

generated. In these studies, the environmental

conditions under which the casts have been exposed

during the ageing – either in vermireactors or in a

controlled systems – have not been defined. Also

the studies were done only at two stages – 15th and

30th day of vermicast generation. Hence, no useful

pointers can be drawn from these studies on the

impact of storage.

The present study, which is perhaps the first

of its kind, explores the changes in the physical,

chemical and biological properties of vermicast that

occur during storage with the objective of finding

conditions that minimize the deterioration in the

fertilizer value of the vermicast. The studies

provide useful pointers on how best to store and

package vermicast.


2.1. Types of storage

The vermicompost used in the present work

was generated from paper waste and the epigeic

species, Eudrilus eugeniae. As paper waste is

almost entirely cellulosic, with only traces of

elements other than C, H, and O, the feed was

spiked with 9% w/w of cow dung in order to

provide NPK and other nutrients in adequate

amounts. The vermicomposting was accomplished

with a high–rate process recently developed by the

author’s group (Gajalakshmi et al., 2012). The

vermireactors were fabricated with aluminum

sheets and each had a volume of 135 liter (15 cm

height with surface area 150 x 60 cm). The

vermicast was harvested after 30 days. One part of

it was stored in two types of packs: (a) airtight

sealed transparent polyethylene bags of 20 micron

thickness (AUD), and (b) partially sealed nylon

mesh (0.3mm) bags (PUD). Both types of bags

were 25 cm long and 18 cm wide, each capable of

holding half kg of vermicompost. Another part of

the casts was pre-dried for 24 hours at room

temperature (29±4°C) and stored in both airtight

sealed transparent polyethylene bags (APD) and

partially sealed nylon mesh bags (PPD). In each set

36 packs were utilized; overall 144 packs were

studied. All storage was at room temperature

(29±4°C) as this is the temperature at which

vermicast is handled in the region where the

authors work. Three packs of vermicast were taken

once in a week for physical and biochemical

analysis from each storage.

2.2. Analysis

The physical properties of vermicast such as

bulk density, particle density and water holding

171

capacity (WHC) were determined by the standard

methods outlined by Bashour and Sayegh (2007)

and Margesin and Schinner (2005). The total

porosity and water filled pore space (WFPS) were

calculated from the particle and bulk density values

of the respective samples (Carter and Gregorich,

2008). Electrical conductivity (EC) and pH were

measured with sample suspension prepared with

distilled water (1:2, w/v) (Bashour and Sayegh,

2007). Total organic carbon (Corg) was determined

following modified dichromate redox method

(Heanes, 1984). Dissolved organic carbon (Cdis)

was extracted in 0.5 M K2SO4 solution (1:10, w/v)

and determined by the dichromate redox method

(Jenkinson et al., 2004). The total nitrogen (Ntot)

was determined by modified Kjeldahl method


digester and distillation units. Inorganic nitrogen

was extracted from moist samples with 2M KCl

solutions (1:10, w/v) followed by determination of

ammonium (NH4+-N) and nitrate (NO3

--N) content

in the suspensions by modified indophenol blue and

Devarda’s alloy method respectively (Jones, 2001).

Extractable potassium (Kext), calcium (Caext)

and sodium (Naext) were determined using a flame

photometer (Elico™ CL378) after extraction with

neutral 1N ammonium acetate solution. Extractable

phosphorus (Pext) was determined according to the

ammonium molybdate-ascorbic acid method

(Knudsen and Beegle, 1988) after extracting with

Mehlich 3 extraction solution (1:25, w/v) (Mehlich,

1984). Mineral sulfur (SO42-

-S) was extracted with

0.0125M CaCl2 solution (1:4, w/v), and determined

by turbidimetric method described by Bashour and

Sayegh (2007). Dehydrogenase enzyme activity

(DHA) was determined by iodo-nitro-

tetrazoliumchloride reduction method (Mersi and

Sehinner, 1991). β-glucosidase (BGA), alkaline

phosphatase (APA) and arylsulphatase (ASA)

enzymes activities were assayed by p-nitrophenol

method as described by Eivazi and Tabatabai

(1977, 1988) and Tabatabai and Bremner (1970).

Cellulase (CEA) activity was assayed by

determination of the reducing sugars released after

incubation of samples with carboxymethyl cellulose

sodium salt (Schinner and von Mersi, 1990). Urease

(URA) activity was assessed by incubating samples

with urea followed by determination of NH4+

released in the hydrolysis reaction by steam

distillation method (Tabatabai and Bremner, 1972).

Microbial biomass carbon (Cmic) was determined by

the chloroform fumigation-extraction method

(Jenkinson et al., 2004).

2.3. Processing of data

The experimental findings were statistically

analyzed to assess whether different treatments

exerted significant impact on the properties of

vermicompost over the course of the storage.

Pearson correlation was used to estimate the degree

of association between each of the vermicast

properties studied and their influence over others.

Statistical significance is recognized at p value ≤

0.05. The SPSS windows 16 package (Softonic,

Barcelona, Spain) was used throughout.



The physical properties of vermicast were

significantly effected by pre-drying and storage

(Table 1). Pre-drying reduced the moisture content,

WHC, total porosity, and WFPS, while it increased

the bulk and particle densities of the cast (Figures

1, 2). In the course of 12 weeks, the moisture

content of the cast of PUD and PPD treatments was

reduced by 69.4±0.1 and 62.1±0.6%, and those of

the AUD and APD by 5.7±0.8 and 7.6±0.6%,

respectively. The bulk density increased in the PUD

and PPD storage to the extent of 49.7±0.8 and

45.8±1.7% and in AUD and APD to the extent of

21.8±2.0 and 29.1±0.3%, respectively. Structural

compactness occurring due to water loss in drying

172

Figure 1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding

capacity (d) of un-dried and pre-dried castings stored in airtight sealed bags (AUD and APD respectively)

and un-dried and pre-dried castings stored in partially sealed bags (PUD and PPD respectively), at different

periods of time.

may be the reason for the greater increase in the

bulk density in the PUD and PPD storage. The

WHC also reduced drastically due to this reason in

both PUD and PPD storage. It was reduced by

about 50% in PUD and PPD storage, in comparison

to 14.1±0.3 and 19.2±1.2%, respectively in the

AUD and APD storage. A significant linear

relationship (p<0.001) was found between WHC

and porosity of castings. At the end of the

experiments, the cast in the PUD and PPD storage

showed about 25% reduction in the total porosity

and those in the AUD and APD about 6% and 12%

reduction, respectively. Hydrophilic components of

organic matter, such as polysaccharides, might also

have influenced the WHC of castings (Li et al.,

2007).

The WFPS values increased to the extent of

22.4±3.4 and 32.7±0.5% in AUD and APD storage

and decreased by 18.4±2.0 and 8.5±5.8% in the

PUD and PPD storage, respectively. Reduction in

structural pores during dehydration and

decomposition of organic matter may be the reason

for lower WFPS of the PUD and PPD storage

0

20

40

60

80

0 2 4 6 8 10 12

Mo

istu

re c

on

ten

t %

No. of weeks

a

0.25

0.35

0.45

0.55

0.65

0 2 4 6 8 10 12

Bu

lk d

en

sity

g c

m-3

No. of weeks

b AUD APD

PUD PPD

1.05

1.10

1.15

1.20

1.25

0 2 4 6 8 10 12

Par

ticl

e d

en

sity

g c

m-3

No.of weeks

c

150

250

350

450

0 2 4 6 8 10 12

WH

C %

No.of weeks

d

173

Figure 2. Changes in the total porosity (a), water filled porosity (b), pH (c) and EC (d) of un-dried and pre-

dried castings stored in airtight sealed bags (AUD and APD respectively) and un-dried and pre-dried castings

stored in partially sealed bags (PUD and PPD respectively), at different periods of time.

(Schwärzel et al., 2002; Kechavarzi et al., 2010).

The changes in the WFPS of vermicast may have a

distinct impact on the regulation of hydrological

properties, gas diffusion, microbial colonization,

nutrient mineralization etc. (Gorres et al., 2001;

Schjønning et al., 2011; Yu et al., 2013). The

particle density of cast showed an increasing trend

in all the storage and the maximum of about 9%

increase was observed in the PUD and PPD. The

rate of increase in particle density of casts indicates

the degree of decomposition of organic matter they

contain (Hassink, 1995; Ruhlmann et al., 2006).

Increasing trend was also observed with EC, in

which maximum increase of 33.5±0.1 and

27.1±2.6% was in AUD and APD storage and

15.5±1.0 and 6.8±3.4%, respectively in PUD and

PPD. The castings in all types of storage had pH

close to neutral all the time (6.99 – 7.21); minor

fluctuations occurred due to the production of

organic acids and the release of CO2 during the

microbial decomposition of organic matter (Elvira

et al., 1998; Ahmad and Qazi, 2014).


The chemical properties of cast were

significantly influenced by pre-drying and storage

55

65

75

85

0 2 4 6 8 10 12

Tota

l po

rosi

ty %

No. of weeks

a

15

19

23

27

31

0 2 4 6 8 10 12

WFP

S %

No. of weeks

b AUD APD

PUD PPD

6.9

7.0

7.1

7.2

7.3

0 2 4 6 8 10 12

pH

No.of weeks

c

1.0

1.2

1.4

1.6

0 2 4 6 8 10 12

EC m

mh

os

cm-1

No.of weeks

d

174

Table 1. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on physical properties, EC and pH of vermicast

during the storage.

Treatment Moisture

content

Bulk

density Particle density

Water holding

capacity Total porosity

Water filled

pore space EC pH

Extend of Sealing 5676.8***

6641.1***

113.8***

886.9***

1781.0***

7.738* 5.290

n.s 4.503

n.s

Pre-Treatment 63025.5***

58466.9***

395.8***

8369.3***

17477.4***

6425.2***

2097.3***

0.318 n.s

Extend of Sealing X Pre-

Treatment 1627.7

*** 4155.3

*** 6.362

* 200.0

*** 1932.8

*** 329.73

*** 3.116

n.s 15.17

*

*p<0.05,

**p<0.01,

***p<0.001,


Table 2. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on chemical properties of vermicast during the

storage.

Treatment

Total

organic

carbon

Dissolved

organic

carbon

Total

nitrogen

Ammonium

-nitrogen

Nitrate-

nitrogen

Available

phosphorus

Exchangeable

potassium

Available

sulfur

Exchangeable

calcium

Exchangeable

sodium

Extend of Sealing 5.953* 10.94

* 1518.9

*** 2447.3

*** 336.5

*** 463.0

*** 271.6

*** 258.6

*** 6763.5

*** 2265.4

***


2414.3***

38905.8***

16255.9***

113692.8***

775.4***

392.5***

1043.4***

1746.4***

34.74***


Treatment 0.346

n.s 19.59

** 1053.5

*** 104.3

*** 536.1

*** 36.84

*** 390.9

*** 791.8

*** 7676.5

*** 2464.6

***

*p<0.05,

**p<0.01,

***p<0.001,


Table 3. F values of repeated measures analysis of variance on the effect of extend of sealing and pre-treatment on biochemical properties of vermicast during the

storage.

Treatment Dehydrogenase

activity

Cellulase

activity

β-Glucosidase

activity Urease activity

Alkaline

phosphatase

activity

Arylsulphatase

activity

Microbial

biomass carbon

Extend of Sealing 54.07

*** 19418.5

*** 1393.6

*** 11285.5

*** 4538.0

*** 2654.1

*** 1876.5

***

Pre-Treatment 98894.9

*** 13.06

** 12427.8

*** 4375.5

*** 5307.3

*** 65483.6

*** 4852.6

***


Treatment 544.3***

11275.7***

281.3***

3477.9***

3957.7***

760.1***

4.894 n.s

*p<0.05,

**p<0.01,

***p<0.001,


175

(p < 0.001) (Table 2). Pre-drying reduced the Corg,

Cdis, Ntot, NH4+-N and Pext and increased the NO3

--

N, Kext, SO42-

-S, Caext and Naext content of the cast

significantly. The PUD and PPD storage showed

higher reduction in Corg, Cdis, Ntot, Pext and Kext than

the AUD and APD storage (Figure 3,4). In the case

of Corg, about 8% reduction was observed with

PUD and PPD storage and less than 1.5% in AUD

and APD. The higher reduction of Corg in the PUD

and PPD, which was maximum in the first two

weeks, was probably due to rapid mineralization of

C by aerobic microbes (Franzluebbers et al., 1994).

A few past studies on the ageing of cast of anecics

and endogeics have also reported high C

mineralization in the initial period (Martin, 1991;

Burtelow et al., 1998; Scullion et al., 2003). The

reduction in WFPS with time may be influencing

the extent of C mineralization as the days

progressed because reduction in the water-filled

pores during the storage might be curtailing the

microbial access to the substrate (Hassink et al.,

1993; Gorres et al., 2001). The cast in the PUD and

PPD storage also showed a maximum reduction in

Cdis content: more than 90% in comparison to

71.7±0.6 and 88.0±0.3%, respectively in the AUD

and APD storage. Low C utilizing efficiency of

anaerobic microbial community, which is expected

to dominate in the AUD and APD storage could be

the reason for the lesser reduction of Cdis content in

these types of storage (Søndergaard and Middelboe,

1995; Song et al., 2008).

The Ntot content of the cast reduced to the

extent of 66.9±0.5 and 54.6±0.3% in the PUD and

PPD storage, respectively. There was about 30%

reduction in the Ntot during the first week, probably

due to intense ammonia volatilization. The high

NH4+-N content of fresh castings also supports this

assumption. As the number of days progressed, the

ammonium content of the casts in the PUD and

PPD storage reduced up to 83% due to the intense

nitrification and ammonia volatilization from the

existing ammonium pools. There was a

concomitant increase in nitrate content of the cast

over time: 72.2±0.3 and 60.0±2.0% with the PUD

and PPD storage, respectively by the end of the

study. Previous studies on cast ageing have also

reported rapid exhaustion of most of the ammonium

present in the castings (Lavelle et al., 1992; Aira et

al., 2005; Kawaguchi et al., 2011b). In contrast, the

cast of AUD and APD storage showed reduction in

both NH4+-N and NO3

--N. Anoxic condition created

in AUD and APD storage due to the airtight sealing

might have impeded the nitrification process

(Jouquet et al., 2011; Aguilar et al., 2014), even as

the slow reduction in the mineral nitrogen content

of the casts may be due to microbial immobilization

(Lavelle and Martin, 1992; Decaens et al., 1999).

In all cases, the Pext in cast increased during

the initial week and further storage showed a steady

decline till the end. The high availability of carbon

and nitrogen in fresh cast would have increased the

phosphorus demand and it probably enhanced

phosphatase activity resulting in increased Pext

during the initial week (Parthasarathi and

Ranganathan, 1999; Flegel and Schrader, 2000). As

the number of days progressed, the Pext in the PUD

and PPD storage reduced by 71.5±0.3 and

73.9±1.5%. In AUD and APD it was reduced by

68.0±1.2 and 61.9±1.6%, respectively. The Kext in

the casts also increased in the initial week, but

further storage reduced it in all the treatments. This

was particularly pronounced in the PUD and PPD

when the reduction in Kext was 16.2±2.1 and

27.5±1.6%; in comparison to 5.4±2.0 and 1.6±2.3%

in AUD and APD, respectively. Similarly the SO42-

-

S content of the cast showed about 20% increase

during the initial week, while further storage

decreased it. The increase in the C/S ratio of the

cast to above 600 that occurred during this period

may be attributed to high immobilization of

available sulfur (Tabatabai and Chae, 1991; Reddy

et al., 2002). Throughout the experiment, the Caext

in the cast fluctuated in the AUD and APD storage.

Increased solubility of organic carbon and

increased competition between the cations for the

negatively charged sites due to increased levels of

Fe and Mn under reducing conditions may be the

reasons for this fluctuation (Wolt, 1994; Phillips

176

Figure 3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c), ammonium

nitrogen (d), nitrate nitrogen (e) and available phosphorus (f) content of un-dried and pre-dried castings

stored in airtight sealed bags (AUD and APD respectively) and un-dried and pre-dried castings stored in

partially sealed bags (PUD and PPD respectively), at different periods of time.

250

260

270

280

290

0 2 4 6 8 10 12

Org

anic

car

bo

n m

g g-1

No. of weeks

a

0.0

2.0

4.0

6.0

8.0

0 2 4 6 8 10 12

DO

C m

g g-1

No. of weeks

b AUD APD

PUD PPD

3

6

9

12

15

0 2 4 6 8 10 12

Tota

l Nit

roge

n m

g g-1

No. of weeks

c

0

100

200

300

400

0 2 4 6 8 10 12

Am

mo

niu

m µ

g g-1

No. of weeks

d

0

100

200

300

0 2 4 6 8 10 12

Nit

rate

µg

g-1

No.of weeks

e

0

40

80

120

160

200

0 2 4 6 8 10 12

Ph

osp

ho

rus

µg

g-1

No.of weeks

f

177

Figure 4. Changes in the exchangeable form of potassium (a), sulfur (b), calcium (c) and sodium (d) content

of un-dried and pre-dried castings stored in airtight sealed bags (AUD and APD respectively) and un-dried

and pre-dried castings stored in partially sealed bags (PUD and PPD respectively), at different periods of

time.

and Greenway, 1998). The Caext in cast of PUD and

PPD storage steadily declined till the end, while

Naext declined to the extent of 11.1±0.8 and

31.1±0.1% in the PUD and PPD storage and

19.0±0.6 and 23.2±0.5%, respectively in AUD and

APD.

3.3. Biochemical properties

Pre-drying of the casts had strong influence

on the enzyme activities (Table 3). Except URA,

the activities of all other enzymes assayed – DHA,

CEA, BGA, APA and ASA – initially increased in

the pre-dried cast before declining. The extent of

this change varied with the type of storage (Figure

5). As much as 82 and 77% increase in DHA was

recorded in the first few weeks of AUD and APD

storage, but further storage reduced the DHA

activity to only 20.9±4.4 and 5.0±4.0%,

respectively. In the case of PUD and PPD storage,

DHA activity increased during the first week, and

then declined to the extent of 89.0±0.3 and

97.6±0.2%, respectively. This trend may be due to

enhanced growth of facultative anaerobic

microorganisms caused by exhaustion of oxygen in

10

12

14

16

18

0 2 4 6 8 10 12

Po

tass

ium

µg

g-1

No. of weeks

a

0

200

400

600

800

0 2 4 6 8 10 12

Sulf

ate

µg

g-1

No. of weeks

b AUD APD

PUD PPD

60

70

80

90

100

0 2 4 6 8 10 12

Cal

ciu

m µ

g g-1

No.of weeks

c

3.5

4.5

5.5

6.5

0 2 4 6 8 10 12

Sod

ium

µg

g-1

No.of weeks

d

178

Figure 5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline

phosphatase (e) and arylsulphatase (f) enzymes activity of un-dried and pre-dried castings stored in airtight

sealed bags (AUD and APD respectively) and un-dried and pre-dried castings stored in partially sealed bags

(PUD and PPD respectively), at different periods of time.

0

30

60

90

120

0 2 4 6 8 10 12

De

nh

ydro

gen

ase

µg

INT

g-1 2

h-1

No. of weeks

a

0

20

40

60

80

0 2 4 6 8 10 12

β-G

luco

sid

ase

act

ivit

y µ

g P

NG

g-1

h-1

No. of weeks

b AUD APD

PUD PPD

0

2

4

6

8

0 2 4 6 8 10 12

Ce

llula

se a

ctiv

ty µ

g C

MC

g-1

24

h-1

No. of weeks

c

0

40

80

120

160

200

0 2 4 6 8 10 12

Ure

ase

act

ivty

µg

NH

4-N

g-1

2 h

-1

No. of weeks

d

0

150

300

450

600

0 2 4 6 8 10 12

Ph

osp

hat

ase

act

ivit

y µ

g P

NP

g-1

h-1

No.of weeks

e

0

30

60

90

120

0 2 4 6 8 10 12

Ary

lsu

lph

atas

e µ

g P

NS

g-1 h

-1

No.of weeks

f

179

Figure 6. Changes in the microbial biomass carbon

content of un-dried and pre-dried castings stored in

airtight sealed bags (AUD and APD respectively)

and un-dried and pre-dried castings stored in

partially sealed bags (PUD and PPD respectively),

at different periods of time.

these types of storage (Stepniewska et al., 1990;

Brzezinâska et al., 1998). Further storage reduced

the DHA activity, possibly due to subsequent

decline in the availability of nutrients.

The BGA activity of the casts increased to

about 73% in the PUD and PPD storage by the

second week. Then there was a steady decline till it

fell to 48.8±0.2 and 49.5±1.2%, respectively at the

end. Similar trend was observed with AUD and

APD storage, in which there was about 67%

increase in BGA activity during the second week,

and 62.4±1.0 and 69.0±0.6% reduction by the

twelfth week. The CEA activity of castings also

increased with PUD, PPD and APD storage during

the initial week, but then fell as much as sevenfold;

those of AUD reduced ninefold. During the initial

week, more than four times increase in APA

activity was observed with the PUD and PPD

storage, and about twice with AUD and APD.

Further storage, reduced the APA activity in all the

cases. In the AUD and APD storage, the reduction

in APA activity was 40.8±1.1 and 47.1±1.2% and

in PUD and PPD storage it was 61.3±1.0 and

71.4±1.9%, respectively.

During the first week, there was a slight

increase in ASA activity with AUD and APD

storage. In the case of PUD and PPD storage, the

ASA activity increased up to third and fourth

weeks, respectively. Further storage showed a

drastic reduction in the ASA activity with all types

of storage: 93.3±0.4, 96.8±0.3, 97.9±0.2, and

99.4±0.1%, in AUD, APD, PUD and PPD,

respectively. Fresh casts had the highest URA

activity, which was reduced to 83.1±1.0, 89.7±0.3,

95.1±0.3, and 94.8±0.3% in AUD, APD, PUD and

PPD, respectively at the end.

In summary, except URA, the activities of all

other enzymes first rose in the initial weeks

possibly due to high availability of nutrients and

physical conditions favorable for aerobic microbial

growth (Allison et al., 2007). Subsequent decline in

the nutrient content (Sinsabaugh et al., 2005; Yao

et al., 2009), moisture content (Poll et al., 2006)

and availability of oxygen (Kang and Freeman,

1999; Xiao-Chang and Qin, 2006) with different

types of storage may have contributed to the

subsequent decline in the enzyme activities.

Pre-drying increased the Cmic content of the

casts significantly; it was 16.5±0.7% higher than

the fresh ones (Figure 6). During the first week,

Cmic increased to 20% in the PUD and PPD storage,

probably due to the high availability of nutrients

which might have promoted high microbial activity

during this period. Further storage led to reduction

across the board: 70.0±0.6, 78.2±0.7, 93.8±0.6,

96.3±0.2%, in AUD, APD, PUD, and PPD storage,

respectively. Subsequent decline in the availability

of nutrients and the moisture content in PUD and

PPD storage may probably be the reason for the

decline in Cmic. In the case of AUD and APD, the

reduction in Cmic may be attributed to the shift of

aerobic microbial groups to anaerobes due to

induced anoxic condition and which has very low C

0

5

10

15

20

0 2 4 6 8 10 12

Bio

mas

s ca

rbo

n m

g g-1

No.of weeks

AUD APD

PUD PPD

180

utilizing efficiency than the former (Song et al.,

2008).

4. Conclusions

A 3-month long study has been described on

the effect of storage on the fertilizer value of

vermicast. In what is arguably the first study of its

kind, several physical, chemical, and biological

attributes of the vermicast as stored with or without

pre-drying, and with or without airtight

containment, were assayed at 7–day intervals. The

manner of storage was seen to influence the plant-

friendly attributes of vermicast in a strong fashion.

Airtight storage after pre-drying was the most

beneficial, followed by airtight storage of the fresh,

undried, vermicast. In partially sealed storage there

was significantly more rapid deterioration of the

beneficial attributes than in airtight storage.

Interestingly, whereas 24-hr pre-drying before

airtight storage was helpful in retaining the plant-

friendly attributes of the vermicast for longer than

fresh-airtight storage, pre-drying before partially

sealed storage had the opposite effect. Apparently,

partially sealed storage added to the water loss that

had already occurred during the pre-drying, and

brought the water content below a level that was

needed to support biological activity within the

vermicast matrix. This indicates that a certain level

of water content is most appropriate for retaining

the microbiological and enzyme activities of the

vermicast; and the presence of water above or

below that level hastens the cast’s ageing. Further

work should be aimed at determining the most

beneficial water levels and how best to retain them.

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4:44

EFFECT OF PRE-DRYING AND EXTENT OF SEALING

ON THE PROPERTIES OF VERMICAST GENERATED

FROM THE NEEM LEAVES DURING STORAGE

Chapter

12

185



CChhaapptteerr 1122

Effect of pre-drying and extent of sealing on the properties of

vermicast generated from the neem leaves during storage

Abstract

In chapter 11, the effect of pre-drying and extent of sealing on the properties of vermicast generated from

paper waste was studied. To understand the impact of these storage practices on the properties of phytomass-

based vermicompost, a similar attempt has been made with the vermicast generated from the neem leaves. In

the present work, neem vermicast was packed in airtight and partially sealed bags with and without pre-

drying for 24 hours and the changes in the physical, chemical and biological characteristics of castings

during storage were monitored for 3 months at 7–day intervals. The storage of vermicast as explored in this

study showed significant differential influence on the properties of vermicast. Reduction in plant available

nutrient and enzyme activity along with changes in the physical properties of castings stored in partially

sealed bags indicate that their beneficial impact on plant growth may get reduced than the castings stored in

airtight bags. Even though, pre-drying of vermicast also reduced some of the beneficial properties of

vermicast during the initial period of storage, this process is recommended before storage as it has prevented

the structural disintegration of castings during storage.

1. Introduction

Enhanced physical, chemical properties and

microbial activity of soil with exogenous organic

amendment such as animal manure and compost

has been reported in numerous studies (Doan et al.,

2013; Ngo et al., 2013). In particular,

vermicompost has received renewed attention in

recent years due to its several plant-friendly

attributes. Application of vermicompost has shown

enhanced physical properties of soil such as

increase in water and nutrient storage capacity,

infiltration and aeration and resistance to

compaction and erosion (Edwards et al., 2004).

Since, most of the nutrients present in the vermicast

are in plant-available form in addition to the

presence of plant growth regulators such as auxins,

gibberellins, cytokinins produced by

microorganisms through interaction with

earthworm (Abbasi and Ramasamy, 1999; Atiyeh et

al., 2001) it possesses remarkable plant growth-

promoting potential on wide range of plants

(Gajalakshmi and Abbasi, 2004; Edwards, 2004).

Studies on a variety of crops such as cereals,

legumes, vegetables, ornamental, medicinal and

flowering plants assessed in both greenhouse and

field studies have confirmed that vermicast,

whether used as soil additives or as components of

plant growth container media, have significant

beneficial effects on seed germination, plant growth

and overall productivity (Edwards, 2004).

Development of disease resistance to a wide range

of plant pathogens is also reported with

vermicompost application (Sahni et al., 2008;

Szczech, 1999).

In the recent years due to the growing interest

of commercial sectors in vermicast, there is the

development of many large-scale commercial

vermicomposting units. In addition, many agro-

186

based industries throughout India, have adopted

vermicast production as part of their commercial

activities. Many thousands of tons of vermicast

produced from these units are supplied throughout

the country in packs which are either fully airtight

or partially sealed polyethylene bags. The vermicast

is packed after drying in shade or sometimes

without drying. There are no specified packing and

storage guidelines for vermicast. The physical,

chemical and biological properties of vermicompost

are subject to change during the storage which is

primary importance in the regulation of soil fertility

and plant growth promotion. Investigating the

effect of storage on vermicast is of great

importance with respect to understanding the

optimum age and storage condition for its best

utilization. The present work is an attempt towards

formulating packing guidelines for storing

vermicast.


2.1. Experimental set up


Pondicherry University campus and its vicinity and

processed with Eudrilus eugeniae, an epigeic

earthworm species. After 30 days, the vermicast

was harvested. In one set of experiment, the

harvested fresh castings was stored in two types of

packing bags: (a) airtight sealed transparent

polyethylene bags of 20 micron thickness, and (b)

partially sealed nylon mesh (0.3mm) bags. The

second set of experiments comprised pre-dried

vermicast at room temperature for 24 hours and

stored in both types of packs. Both types of bags

were 25 cm long and 18 cm wide, each capable of

holding half kg of vermicompost. In this study,

there were 36 packs for each treatment, so totally

144 packs were prepared. All the packs of

vermicast were stored at room temperature in order

to imitate the general way of storage of

vermicompost in commercial sectors. Three packs

of vermicast from the all the treatments were

sampled once in a week for physical, chemical and

biochemical analysis.




2.3. Data analysis

Data was analyzed using repeated analysis of

variance at the 0.01% level for which with/without

pre-drying and type of sealing were fixed as

between-subject factors and storage period was

fixed as within-subject factor. Relationships

between different vermicast properties and their

influence over others were described and tested by

Pearson correlation coefficients. All analyses were

performed using SPSS 16 package.



The change in the physical properties of

vermicast with different treatments is given in

Figures 1 and 2. During storage, castings stored in

partially sealed bags showed about 70% loss in

moisture content. In the airtight bags, a maximum

of 7.8% of water loss was recorded in 12 weeks of

storage. The changes in the moisture content of the

castings during air drying and storage has promoted

compactness in the structure resulting in the

increase in bulk density. An 11.5±1.6% increase in

bulk density was recorded during air-drying. The

bulk density of the castings stored in partially

sealed bags increased more sharply between the

first 42 days. At the end of 12 weeks, the increase

in bulk density was 52.0±0.2 and 45.3±1.9% with

un-dried and pre-dried castings, respectively. The

castings stored in airtight bags showed 21.6±3.8

and 25.9±2.6% increase in bulk density with un-

dried and pre-dried castings, respectively.

For organic matter, the particle density

depends on the degree of decomposition, and

187

Figure 1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding capacity (d), of

un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried

castings stored in partially sealed bags (T3 and T4 respectively), at different periods of time.

ranges between 1.0 and 1.5 g cm-3

(Hassink, 1995;

Ruhlmann et al., 2006). In the present study, the

particle density of castings increased significantly

till the end of the experimental period irrespective

of the treatments. Increase in particle density was

higher in castings stored in partially sealed bags

than the airtight bags and it was ranging from 1.063

to 1.736 g cm-3

. In the case of water-holding

capacity (WHC) and total porosity, there was no

significant difference observed during the pre-

drying. The castings stored in the partially sealed

bags showed higher reduction of about 45% in

WHC in 12 weeks; whereas it was 19.0±0.8 and

34.3±1.2% with un-dried and pre-dried castings in

airtight bags. In this study, a significant linear

relationship (p<0.001) was found between WHC

and porosity of castings (Table 1). It indicates that

the decrease in WHC of castings during the storage

was likely due to the reduction of pore space in the

castings (Chaudhuri et al., 2009). Moreover, a

decrease in hydrophilic components of organic

matter e.g. polysaccharides (Piccolo and Mbagwu,

1999) during the storage might be limiting the

available surface area that absorbs water, resulting

in the decline in water holding capacity of castings

(Li et al., 2007).

The total porosity of castings decreased in all

the treatments. The castings stored in partially

sealed bags showed 10.3±0.5 and 11.8±1.0%

0

20

40

60

80

0 2 4 6 8 10 12

Mo

istu

re c

on

ten

t %

No. of weeks

a

0.25

0.35

0.45

0.55

0.65

0 2 4 6 8 10 12

Bu

lk d

en

sity

g c

m-3

No. of weeks

b T1 T2 T3 T4

0.9

1.1

1.3

1.5

1.7

1.9

0 2 4 6 8 10 12

Par

ticl

e d

en

sity

g c

m-3

No. of weeks

c

300

400

500

600

700

800

0 2 4 6 8 10 12

WH

C %

No. of weeks

d

188

Figure 2. Changes in the total porosity (a), water filled porosity (b), pH (c), and EC (d), of un-dried and pre-dried

castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried castings stored in partially

sealed bags (T3 and T4 respectively), at different periods of time.

reduction in total porosity with un-dried and pre-

dried castings, respectively. The change in airtight

bags was only between 2.9±1.4 to 6.1±1.4%. The

water filled pore space (WFPS) of the castings was

reduced by 23.9±1.2% during pre-drying. In case of

airtight bags it was showing an increasing trend

with 20.7±6.4 and 24.8±4.3% increase in un-dried

and pre-dried vermicast respectively. In the

partially sealed bags, a steep reduction in WFPS

was observed as the number of days progressed and

it was 46.5±1.0% and 35.7±1.6% with un-dried and

pre-dried vermicast. Higher reduction in WFPS in

the partially sealed bags may be due to loss of

structural pores during dehydration and

decomposition (Schwärzel et al., 2002; Kechavarzi

et al., 2010). The changes in the WFPS of

vermicast have distinct impact on the other

physical, chemical and biological properties; WFPS

is of primary importance in the regulation of

hydrological properties, gas diffusion, microbial

colonization, nutrient mineralization etc. (Gorres et

al., 2001; Schjønning et al., 2011).

The pH of fresh vermicast was 6.71±0.01 on

the day of the deposition and it reduced to 6.11-

6.48 at the end of the experiment. The castings

stored in the airtight bags showed little fluctuation

in the pH, whereas the castings in partially sealed

bags exhibited a steady decline throughout the

experimental period. The reduction in pH during

63

67

71

75

79

0 2 4 6 8 10 12

Tota

l po

rosi

ty %

No. of weeks

a

10

20

30

40

0 2 4 6 8 10 12

WFP

S %

No. of weeks

b T1 T2

T3 T4

6.0

6.2

6.4

6.6

6.8

0 2 4 6 8 10 12

pH

No. of weeks

c

1.2

1.5

1.8

2.1

0 2 4 6 8 10 12

EC m

mh

os

cm -1

No. of weeks

d

189


during the storage.

Treatment Moisture

content

Bulk

density Particle density

Water holding


Water filled

pore space EC pH


5.820***

2.487***

4.470***

72.95***

3.050***

111.1***

29.68**


5.372***

2.872***

1.534***

45.22***

871.6***

9913.9***

953.1***


Treatment 3.993

*** 497.1

*** 984.0

*** 687.7

*** 1.664

n.s 195.5

*** 672.5

*** 603.8

***

*p<0.05,

**p<0.01,

***p<0.001,



storage.

Treatment

Total

organic

carbon

Dissolved

organic

carbon

Total

nitrogen

Ammonium-

nitrogen

Nitrate-

nitrogen

Available

phosphorus

Exchangeable

potassium

Available

sulfur

Exchangeable

calcium

Exchangeable

sodium


1.576***

9.188***

3.363***

2.714***

257.2***

3.342***

9.410***

6.058***

1.114***


629.4***

2.363***

1.064***

1.067***

2.836***

141.0***

1.385***

35.85***

4.199***

Extend of Sealing X

Pre-Treatment 211.0

*** 532.4

*** 362.4

*** 4.556

*** 49.25

*** 13.71

** 622.2

*** 901.1

*** 888.3

*** 5.109

***

*p<0.05,

**p<0.01,

***p<0.001,



storage.


activity

Cellulase

activity

β-Glucosidase


Alkaline

phosphatase

activity

Arylsulphatase

activity

Microbial

biomass carbon


*** 360.3

*** 3.454

*** 5.675

*** 3.341

*** 4.301

*** 2.209

***

Pre-Treatment 90.71

*** 1.930

*** 8.834

*** 1.893

*** 1.890

*** 9.446

*** 2.766

***


Treatment 155.3***

2.257***

4.569***

1.597 n.s

7.252***

8.230***

4.699***

*p<0.05,

**p<0.01,

***p<0.001,


190

the storage may be due to production of carbon

dioxide and organic acid during the microbial

decomposition of organic matter (Elvira et al.,

1998). The electrical conductivity (EC) of fresh

vermicast was 1.44±0.03 mmhos cm-1

on the day of

deposition and it increased till the end of the

experiment with castings of airtight bags. In the

case of partially sealed bags, an increasing trend in

EC was observed during the first three weeks and

further storage reduced the EC till end of the

experimental period.


There was a significant influence (p < 0.001)

on the chemical properties of vermicast with pre-

drying and storage (Figures 3 and 4; Table 2). Pre-

drying of vermicast showed reduction in organic

carbon and nitrogen content. During storage, the

castings stored in the partially sealed bags showed

constant reduction of C content and it was 6.5±0.1

and 7.3±0.2% for un-dried and pre-dried castings

respectively. The reduction of C content in partially

sealed bags was due to the rapid mineralization of

C by aerobic microbes in this treatment

(Franzluebbers et al., 1994). In this study, the C

loss was high during the initial week of storage and

this finding is similar to the report of Martin

(1991), Burtelow et al. (1998) and Scullion et al.

(2003). The reduction in C loss/mineralization as

the days progressed might be the reason for

reduction in WFPS which holds a greater

proportion of bacteria (Hassink et al., 1993). The

reduction in this pore size during the storage

curtails the microbial access to the substrate

(Gorres et al., 2001). The DOC in the vermicast

showed differential response to different treatments

(Figure 3b). The castings stored in partially sealed

bags showed a steady decline in DOC content

during the entire period of storage. In the course of

12 weeks storage, reduction in DOC was 86.0±0.2

and 88.4±0.1% with un-dried and pre-dried

castings, respectively. The castings stored in the

airtight bags showed maximum DOC content

throughout the experiment and the reason may be

the shift in aerobic condition in this treatment

which has reduced the C utilizing efficiency of the

microbial community (Søndergaard and Middelboe,

1995; Song et al., 2008). In this treatment, an

increasing trend in DOC content was observed

during second to fifth weeks, and further storage

decreased the DOC content by 44.3±0.8 and

37.6±1.0% with pre-dried and un-dried vermicast,

respectively.

The concentration of total nitrogen decreased

in all the treatments (Figure 3c) and the changes

were significantly different among the treatments

(p<0.001) (Table 2). The N concentration in un-

dried and pre-dried castings stored in partially

sealed bags reduced to 32.8±0.2 and 37.2±0.2% in

the course of 84 days storage. In these treatments

around 30% of nitrogen loss with pre-dried and un-

dried castings was observed in the first three weeks

of storage. The results reflected the high rate of

nitrogen loss in the castings during the initial weeks

of storage and it was likely due to the intense

ammonia volatilization. This result is in contrast to

the report of other field studies in which the total N

of castings were rather constant during the entire

ageing process (Decaëns et al., 1999; Jiménez and

Decaëns, 2004). These studies (Decaëns et al.,

1999; Jiménez and Decaëns, 2004) were conducted

with castings in the size of about 6 cm diameter

produced by the large anecic earthworm,

Martiodrilus sp. The larger size of the castings may

impede aeration and prevent the ammonium from

volatilization within the aggregates, whereas

castings that was stored in the present study was <

2 mm dia. Moreover, in the above cited studies the

pH of castings was 4.5 and 5 and the ammonia

volatilization completely stops at this pH (Hartung

and Phillips, 1994).

The concentration of NH4+ and NO3 in the

fresh castings was 669.4±3.9 and 57.5±0.9 mg kg-1

on the day of deposition. The high NH4+ content in

the castings may be due to high mineralization of

substrate during the gut passage and the addition of

urine in the posterior part of the digestive tract

191

Figure 3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c), ammonium nitrogen

(d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and pre-dried castings stored in airtight sealed bags (T1

and T2 respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at

different periods of time.

340

360

380

400

0 2 4 6 8 10 12

Org

anic

car

bo

n m

g g-1

No. of weeks

a

0.0

1.0

2.0

3.0

4.0

5.0

0 2 4 6 8 10 12

DO

C m

g g-1

No. of weeks

b T1 T2 T3 T4

10

14

18

22

26

30

0 2 4 6 8 10 12

Tota

l nit

roge

n m

g g-1

No. of weeks

c

0

200

400

600

800

0 2 4 6 8 10 12

Am

mo

niu

m µ

g g-1

No. of weeks

d

0

200

400

600

0 2 4 6 8 10 12

Nit

rate

µg

g-1

No. of weeks

e

0

100

200

300

0 2 4 6 8 10 12

Ph

osp

ho

rus

µg

g-1

No. of weeks

f

192

Figure 4. Changes in the extractable potassium (a), sulfur (b), calcium (c) and sodium (d) of un-dried and pre-dried



(Decaëns et al., 1999). Pre-drying of castings had

showed 35.5±0.2% of reduction in the initial NH4+

concentration. The decease of NH4+ in the castings

is reported to be reflected by increase in NO3-

concentration due to the nitrification of existing

ammonium pools (Lavelle and Martin, 1992;

McInerney and Bolger, 2000). Similar high

nitrification process was observed in the castings of

all the treatments during the first week of storage

and then it decreased progressively. Amongst

different treatments, maximum nitrification was

observed in castings stored in partially sealed bags.

In this treatment, the un-dried and pre-dried

castings showed 38.4±0.3 and 24.7±0.7% reduction

in NH4+ concentration at the first week, and then

81.7±0.7 and 78.5±0.5% reduction at the end of 84

days, respectively. The results are agreeing with

other findings of N dynamics during the cast

ageing. The castings of geophagous tropical

earthworm Pontoscolex corethrurus had reported

41% rapid nitrification during 16.5 days of

incubation (Lavelle et al., 1992). The casts of large

endogeic species Aporrectodea caliginosa has

shown up to 92.3% of NH4+ reduction during the 60

days laboratory incubation study (Aira et al., 2005).

Kawaguchi et al. (2011) reported that castings of

epigeic earthworm species Metaphire hilgendorfi

showed up to 30% NH4+ reduction during the 56

5

9

13

17

21

25

0 2 4 6 8 10 12

Po

tass

ium

µg

g-1

No. of weeks

a

0

200

400

600

800

0 2 4 6 8 10 12

Sulf

ate

µg

g-1

No. of weeks

b T1 T2

T3 T4

100

200

300

400

0 2 4 6 8 10 12

Cal

ciu

m µ

g g-1

No.of weeks

c

2.0

3.0

4.0

5.0

6.0

0 2 4 6 8 10 12

Sod

ium

µg

g-1

No.of weeks

d

193

days incubation.

The castings stored in airtight bags showed

30.2±0.8 and 35.1±0.5% of NH4+ reduction with

un-dried and pre-dried castings respectively. It can

be noted that the NH4+ reduction of up to

14.2±0.7% was observed in the first week itself and

then the rate of NH4+ loss largely reduced. Since,

nitrification process require oxic condition (Jouquet

et al., 2011), the anoxic condition created in airtight

bags after a week of storage would have impeded

this process. However, there was a slow reduction

in NH4+ observed throughout the storage process

along with gradual reduction in NO3-, certainly as

the result of denitrification and microbial

immobilization (Lavelle and Martin, 1992; Decaëns

et al., 1999). After 42 days of storage, fluctuation in

mineral N greatly reduced and this may be due to

reduction in microbial activity which in turn is due

to excess dryness. Similar findings were reported

with castings of endogeic earthworm Milsonia

anomala during the ageing process (Martin, 1991;

Lavelle and Martin, 1992).

In both airtight and partially bags increase in

labile P was observed in the initial week of the

storage and then there was a steady decline till the

end. Largely available nutrient pool and high

moisture content of fresh castings would have

enhanced phosphatase activity; the resultant higher

mineralization increased the fraction of labile P

pool in the vermicast during this period

(Parthasarathi and Ranganathan, 1999; Flegel and

Schrader, 2000). A similar result was also reported

with the castings of same earthworm species with

press-mud as feed and other species such as

Lampito mauritii and Martodrilus carimaguensis

during the ageing process (Satchell and Martin,

1984; Parthasarathi and Ranganathan, 1999;

Jimenez et al., 2003). Further storage decreased the

labile P content and there was no significant

difference between different treatments.

Exchangeable K content in the fresh castings

increased to 26.7±1.9% during the pre-drying and

this may be due to increase in the microbial

biomass during this period. Similarly, an increase in

K release during the drying was reported in soils

(Gupta And Rorison, 1974) and wet meadows

(Johnston et al., 1995). However, other authors

observed a decreased K release after soil drying

(Koerselman et al., 1993; Lamers et al., 1998), or

an unaffected K release (Grootjans et al., 1986).

During storage, the exchangeable K content

declined in all the treatments. The decrease in

exchangeable K was much higher in the casting of

partially sealed bags than the airtight bags (Figure

4a). Amongst all the treatments, the castings stored

in the partially sealed bags showed maximum of

43.5±0.6 and 67.6±0.3% reduction with un-dried

and pre-dried castings, respectively; whereas, the

decrease in airtight bags was 5.5±1.6 and

26.4±0.8% with the un-dried and pre-dried castings

during 84 days storage. The differential response of

exchangeable K during the storage with different

treatments could not be attributed to any particular

reason as relevant reports on the dynamics of K in

the vermicast and even on the biogeochemistry of

K in soils are very limited.

The sulfate content of vermicast increased by

7.8±1.1% during the pre-drying and the

mineralization of S continued to different extent

with different treatments (Figure 4b). The increase

in labile S pool in the vermicast during the pre-

drying was probably due to the breakdown of very

labile sulfate esters (Li et al., 2001). Increase in

arylsulphatase activity also indicates the increase in

microbial mineralization of sulphur during pre-

drying. During storage, an increase in labile S was

recorded in the initial weeks of storage with all the

treatments. Further storage showed reduction of

55.0±1.0 and 41.9±1.0% in S content with un-dried

and pre-dried castings stored in partially sealed

bags. In the case of airtight bags, the reduction was

about 30%. The castings stored in the airtight bags

showed fluctuation in both Ca and Na content

during the entire period of storage (Figures 4c and

4d). At the end of the experimental period, the un-

dried and pre-dried castings stored in airtight bags

showed 40.8±2.2 and 32.6±1.6% increase in Ca

194

content; whereas, in the partially sealed bags,

7.6±4.8 and 19.2±0.4% reduction in Ca content was

observed with un-dried and pre-dried castings,

respectively. The Na content of the un-dried

castings stored in the airtight bags showed increase

of 13.6±1.1% at twelfth week. Although, the pre-

dried castings showed a similar trend of results, at

the end of the experimental period about 30%

reduction in Na content was observed. In the case

of partially sealed bags, the Na content of the

castings was reduced about 50% during the storage.

The increase in Ca and Na content in the airtight

bags may be attributed to increased solubility of

organic carbon, and increased competition between

the cations for the negatively charged sites due to

increased levels of Fe and Mn under reducing

conditions (Wolt, 1994; Phillips and Greenway,

1998).


Typical changes of enzyme activity of

vermicast during the pre-drying and storage with

different treatments is shown in Figure 5. The

changes in enzyme activity were significantly

different among the treatments (p<0.001; Table 3).

It can be seen that dehydrogenase activity increased

to 22.4±0.4% during the pre-drying and the

increasing trend in this enzyme activity was

continued up to 42 and 49 days with un-dried and

pre-dried castings stored in airtight bags,

respectively. The increase in dehydrogenase

activity was more distinct in the un-dried castings

stored in airtight bags with the maximum of

74.4±0.3% higher activity in comparison to the

fresh castings. Whereas, 64.9±0.04% increase in

dehydrogenase activity was observed in the pre-

dried castings of similar treatment. After this, there

was a steady decline in dehydrogenase activity in

the airtight bags till the end of the experiment. The

castings stored in the partially sealed bags, showed

about 17% increase in dehydrogenase activity

during the first week and further storage decreased

the enzyme activity. In this treatment, about 85%

reduction in dehydrogenase activity was observed.

During storage, shift in aerobic to anaerobic

condition in airtight bags could be the reason for

the higher dehydrogenase activity (Brzezinâska et

al., 1998). Many studies on soil enzyme activity

reported similar high dehydrogenase activities in

anoxic condition (Stepniewska et al., 1990;

Brzezinâska et al., 1998).

The β-glucosidase activity increased to

34.1±0.5% during pre-drying. During the initial

period of storage, there was an increase in β-

glucosidase activity with both airtight and partially

sealed bags treatment, and the increase was more

distinct in the partially sealed bags. A maximum of

80% increase in enzyme activity was observed in

this treatment during the third week of storage;

whereas it was about 70% increase with airtight

bag. In the case of partially sealed bags, the higher

β-glucosidase activity during the first 3 weeks of

storage can be explained by their oxic condition.

Studies have shown similar changes in β-

glucosidase activity in the wetland soil during

changes in their oxic condition by varying moisture

content (Vo and Kang, 2013). From third week

onwards, a constantly reduced β-glucosidase

activity was observed in all the treatments till the

end of the experiment. Lower β-glucosidase activity

was observed in airtight bags indicating that the

change in the redox states of the soil impedes the β-

glucosidase activity in the vermicast; whereas, in

the partially sealed bag, reduction may be attributed

by reduction of microbial activity and degradation

of this enzyme due to excess dryness (Poll et al.,

2006).

Like other hydrolytic enzymes, the cellulase

activity also increased during the pre-drying

process. In this study, the fresh castings showed

1.34±0.02 µg CMC g-1

24h-1

cellulase activity and

increased by greater than twofold during pre-

drying. The un-dried and pre-dried castings stored

in the airtight bags showed a constant reduction in

cellulase activity and at the end of the twelfth week

reduction was 74.0±2.4 and 86.0±1.0%,

respectively (Figure 5c). Castings stored in the

195

Figure 5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline phosphatase (e) and

arylsulphatase (f) enzymes activity of un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2

respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at different

periods of time.

0

20

40

60

80

100

0 2 4 6 8 10 12

De

nh

ydro

gen

ase

µg

INT

g-1 2

h-1

No. of weeks

a

0

20

40

60

80

100

0 2 4 6 8 10 12

β-G

luco

sid

ase

act

ivit

y µ

g P

NG

g-1

h-1

No. of weeks

b T1 T2

T3 T4

0

1

2

3

4

0 2 4 6 8 10 12

Ce

llula

se a

ctiv

ty µ

g C

MC

g-1

24

h-1

No. of weeks

c

0

40

80

120

160

200

0 2 4 6 8 10 12

Ure

ase

act

ivty

µg

NH

4-N

g-1

2h

-1

No. of weeks

d

0

200

400

600

800

0 2 4 6 8 10 12

Ph

osp

hat

ase

act

ivit

y µ

g P

NP

g-1

h-1

No. of weeks

e

0

10

20

30

40

50

0 2 4 6 8 10 12

Ary

lsu

lph

atas

e a

ctiv

ity

µg

PN

S g-1

h-1

No. of weeks

f

196

partially sealed bags also showed a similar trend, in

which 75.3±1.4 and 89.1±1.3% reduction in

cellulase activity was observed with un-dried and

pre-dried castings, respectively. During storage, the

reduction in the cellulase activity of castings in all

the treatments may be probably due to reduction in

available nutrient such as inorganic N as reported in

soil and plant litter based studies (Sinsabaugh et al.,

2005; Yao et al., 2009).

The casting showed significantly high urease

activity of 180.6±2.2 µg NH4-N g-1

2h-1

on the day

of deposition and reduced to 8.5±1.3% during the

pre-drying. The declining trend of urease activity

was seen in all the treatments (Figure 5d). Amongst

different treatments, the un-dried and pre-dried

castings stored in partially sealed bags showed

fastest reduction in urease activity, where the

reduction was 50.0±0.5 and 54.2±0.4%,

respectively during the second week of storage. At

the end of the experiment, castings stored in these

treatments showed more than 22 and 26 folds

reduction in urease activity in the case of un-dried

and pre-dried castings, respectively. In the case of

castings stored in airtight bags, the reduction rate

was significantly lower than the castings of

partially sealed bags. In this treatment, urease

activity of un-dried and pre-dried castings reduced

to 84.1±0.4 and 88.2±0.3% respectively; it was

about 6 and 8 folds lower than the one day old

castings of respective treatments. The considerable

portion of urea present in the castings as earthworm

urine could be the reason of observed higher urease

activity in fresh vermicast (Edwards, 2004). The

decline in urease activity during the pre-drying and

storage probably happened due to the denaturing of

enzyme (Gould et al., 1973). Similarly, the change

in urease activity of soil during the incubation

under moist and dry condition has been reported in

previous studies (Zornoza et al., 2006; Geisseler et

al., 2011).

Alkaline phosphatase activity of the fresh

vermicast increased during the pre-drying and the

first week storage. As the days of storage

progressed, there was a steady decline in the

phosphatase activity with all the treatments. In the

case of un-dried castings stored in airtight bags,

decline in phosphatase activity was 44.7±1.6 and

36.5±0.3%, respectively; whereas in the partially

sealed bags, maximum reduction of 71.8±0.6% was

observed in the pre-dried castings followed by un-

dried castings which showed 45.4±0.9% reduction.

A possible explanation for the high phosphatase

activity during the pre-drying and first week of

storage may be due to high microbial activity in

vermicast during this period. Increase in labile N

and other nutrient content in the fresh vermicast

probably increased the P demand, a likely

consequence of higher phosphatase activity

stimulated by the active microbial population

(Allison and Vitousek, 2005; Allison et al., 2007).

The increase in labile P content by stimulated high

enzyme activity may have contributed to the slow

decline in phosphatase activity in the continuing

weeks of storage. In the case of castings stored

airtight, reduction in enzyme activity is probably

due to the low P requirement of anaerobic bacteria

and inhibition of phosphatase activity with oxygen

depletion as reported in many soil based studies

(Pulford and Tabatabai, 1988; Hinojosa et al.,

2004).

The castings showed 22.5±0.0 µg PNS g-1

h-1

of arylsulphatase activity on the day of deposition

and pre-drying increased the enzyme activity by

43.4±0.1%. Increase in arylsulphatase activity in

vermicast during the air-drying was consistent with

the reports of Tabatabai and Bremner (1970).

During storage, the castings stored in airtight bags

showed steady decline in enzyme activity from the

initial day of storage. In this treatment, 94.5±0.1

and 97.8±0.1% of reduction in enzyme activity was

observed within un-dried and pre-dried castings,

which was higher than the castings of partially

sealed bag treatments. The decline in arylsulphatase

activity in airtight bags is probably due to the

increase of redox states as reported on waterlogged

soil (Kang and Freeman, 1999; Xiao-Chang and

Qin, 2006). Reduction in arylsulphatase activity in

197



airtight sealed bags (T1 and T2 respectively) and un-

dried and pre-dried castings stored in partially sealed

bags (T3 and T4 respectively), at different periods of

time.

anoxic condition might be the reason for changes in

the microbial community. Moreover, anaerobic

conditions also mobilize some metal ions, notably

Fe2+

, which might have impeded this enzyme

activity in castings stored in the airtight bags

(Pulford and Tabatabai, 1988; Freeman et al.,

1996). In the case of partially sealed bags, un-dried

and pre-dried castings showed increase in enzyme

activity during the initial period of storage, and then

there was a slow reduction in enzyme activity till

the end. In this treatment, a maximum reduction of

85.5±0.1% was observed in the pre-dried castings

followed by 67.1±0.1% in un-dried castings in the

course of 12 weeks of storage.

Microbial biomass carbon in the fresh

vermicast increased by 24.7±0.4% during pre-

drying. Storage of vermicast greatly affected the

microbial biomass C content of castings stored in

both airtight and partially sealed bags (Figure 6).

During the first week of storage, increase in the

microbial biomass C was observed in all the

treatments, and the increase was predominant in the

un-dried castings stored in both airtight and

partially sealed bags. During this period, the un-

dried castings showed 23.02±0.6 and 21.01±0.2%

increase in microbial C with partially sealed and

airtight bags, respectively. Continuing storage of

castings was characterized by reduction in

microbial C in the course of 12 weeks storage.

Castings stored in airtight bags showed 75.50±0.2

and 76.07±0.7% reduction in biomass with un-dried

and pre-dried castings, respectively at the end of the

experiment. Partially sealed bags treatment showed

steady decline in biomass and it was 85.03±0.5 and

94.05±0.2% with un-dried and pre-dried castings.

Although other studies showed reduction

(Parthasarathi and Ranganathan, 1999; Tiunov and

Scheu, 2000; Scullion et al., 2003) or no changes

(Aira et al., 2005, 2010) in biomass during the

initial period of ageing, in the present study higher

microbial activity and biomass was observed during

the first week.

Most of these previous studies are field based

and conducted with castings from the geophagous

earthworm, which are reported to contain a

considerable portion of mineral particles (Lavelle

and Spain, 2003; Blanchart et al., 2004; Jouquet et

al., 2008) and lower nutrient than the castings of

phytophagous worms, epigeic. This lower nutrient

content of the castings of geophagous are subject to

exhaustion very soon by intense microbial activity

(Tiunov and Scheu, 2000; Aira et al., 2005). In the

present study, availability of excess phytomass

attributed abundant nutrient availability in the fresh

vermicast, which would have created hotspot of

microbial activity during the first week of storage

and increased their biomass. The progressive

decline of available nutrients and moisture content

of vermicast was related to the resultant decline in

microbial biomass in further storage (Scheu, 1987).

The higher reduction of microbial C in the airtight

treatment could be due to the elimination of aerobic

microbial groups and shift in microbial community

due to induced anoxic condition. Microbial

communities under anoxic condition has low

energy yield from metabolizing reduced substrates,

leading to low C and other nutrient use efficiency

(Song et al., 2008).

0

5

10

15

20

25

0 2 4 6 8 10 12

Bio

mas

s ca

rbo

n m

g g-1

No.of weeks

T1 T2

T3 T4

198

4. Conclusions

The finding of this study reveals that

physical, chemical and biochemical properties of

vermicast are subject to modification during the

storage, irrespective of different treatments used in

this study. Amongst the treatments, the castings

stored in the partially sealed bags showed a drastic

change in their physical, chemical and enzymatic

properties. The physical properties such as

gravimetric water content, porosity and water-

holding capacity largely reduced whereas the bulk

density and particle density increased maximally in

the partially sealed treatment. These changes can

impede the water availability, oxygen diffusion and

plant root penetration in the field. Although, the

castings stored in airtight bags also showed changes

in these properties, it was less intense. The castings

stored in the partially sealed bags also showed a

higher reduction in total C, N and other nutrients

than the airtight treatment. Results of enzymes

assays and microbial biomass C content in partially

sealed treatment indicate intense aerobic microbial

activity in the initial period of storage which would

have immobilized and stabilized the available

nutrient pool in the organic matter.

Due to the excess dryness of castings stored

in partially sealed bag, nutrients would have highly

stabilized in organic matter and this would impede

the instant nutrient release to plants in the field.

Therefore, the beneficial impact of the vermicast on

the plant growth will be very minimum when stored

in partially sealed bags in comparison to either

fresh vermicast or castings stored in airtight bags.

Although, the beneficial properties reduced in the

castings stored in airtight bags, it was less intense.

Since there is no loss in nutrient due to their airtight

sealing and the physical properties of these castings

also favor the growth of microbes even after

prolonged storage, the disappeared nutrient pool

can be regained once it is applied into the field.

Even though, pre-drying of vermicast reduced some

of the beneficial properties of vermicast, this

practice can be recommended before storage as it

prevents the disintegration of the structure of

castings due to excess moisture. The small pellet

like structure of castings having higher surface area

will be available for microbial activity thereby

enhancing the mineralization of nutrients even after

prolonged storage.

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cellulase and invertase activity in soil: an


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M., Bennicelli, R., Stepniewski, W., 1990.

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30, 53-59.

Szczech, M.M., 1999. Suppressiveness of

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Biochem. 41, 1425-1432.

Zornoza, R., Guerrero, C., Mataix-Solera, J.,

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2134.

EFFECT OF PRE-DRYING AND EXTENT OF SEALING

ON THE PROPERTIES OF VERMICOMPOSTED COW

DUNG DURING STORAGE

Chapter

13

203



CChhaapptteerr 1133

Effect of pre-drying and extent of sealing on the properties of

vermicomposted cow dung during storage

Abstract

In chapters 11 and 12, studies on impact of pretreatment process and packing method on the properties of

vermicast generated from paper waste and neem leaves during storage were reported. In continuation to these

studies, the impact of the storage practices on manure based vermicast have been examined and reported in

this chapter. The changes in the physical, chemical and biochemical characteristics of castings during storage

were monitored for 3 months with weekly samples. The results reveal that the properties of castings get

altered irrespective of the mode of storage explored in this study. The castings stored in the bags which are

not fully airtight showed drastic losses in the plant available nutrient and enzyme activity along with adverse

changes in its physical properties than the castings stored in fully airtight bags. Even though, preprocessing

of vermicast such as drying has reduced some of the beneficial properties of vermicast; it prevented the

disintegration of the structure of castings due to excess moisture during storage.

1. Introduction

As discussed in chapters 11 and 12,

vermicast, whether used as soil additives or as

components of plant growth in container media,

have significant beneficial effects on seed

germination, plant growth and overall plant

productivity (Edwards, 2004; Lazcano, and

Domínguez, 2011). The beneficial aspects

contributed by vermicast on plant growth may be

attributed by various mechanisms, such as a

modification in soil structure, change in water

availability, increase in availability of macro and

micronutrients, stimulation of microbial and

enzymatic activities, or production of plant growth-

promoting materials by microorganisms through

interactions with earthworms (Abbasi and

Ramasamy, 1999; Atiyeh et al., 2001).

All these beneficial mechanisms rely on the

properties of vermicast which is subject to various

changes during storage. As there are no specified

packing and storage guidelines to preserve the

beneficial properties of vermicast during storage,

the present study was conducted. An an attempt has

been taken to explore the changes in the physical,

chemical and biological properties of manure based

vermicast during storage in order to understand the

optimum age and storage condition for its best

utilization.


2.1. Experimental design

Vermicompost was generated from the cow

dung using an epigeic earthworm, Eudrilus

eugeniae. For vermicomposting, 140 litre volume

vermireactors fabricated with aluminum sheet were

employed. Fifty number adult worms per kg of feed

were introduced into 25 kg of cow dung on dry

weight basis and composted for 30 days. In order to

assess the impact of different storage method on

vermicast properties, the harvested vermicast was

stored by following methods: (i) fresh vermicast

stored in airtight bags (T1); (ii) vermicast pre-dried

204

for 24 hours (at room temperature under shade)

stored in airtight bags (T2); (iii) fresh vermicast

stored in partially sealed bags (T3), and (iv)

vermicast pre-dried for 24 hours stored in partially

sealed bags (T4). Each treatment comprised 36

packs of vermicast with 500 g of castings. All the

144 packs were stored at room temperature in order

to mimic the general way of storage of

vermicompost in commercial sectors. Three packs

of vermicast from the all the treatments were

sampled once in a week for physical and

biochemical analysis.




2.3. Data analysis

The influence of different storage methods on

the properties of vermicast were tested through

repeated analysis of variance. Pearson correlation

was also used to estimate the degree of association

between each of the vermicast properties studied

and their influence over others. All statistical

calculations were carried out using SPSS windows

16 package. Differences were considered

significant only when p-values were lower than

0.05.



The physical properties of vermicast changed

significantly during the pre-drying and storage,

irrespective of different treatments used in this

study (Table 1). Pre-drying of vermicast reduced

the moisture content, water-holding capacity

(WHC) and water filled pore space (WFPS) by

11.2±1.3, 18.8±2.7, 11.48±1.42%, respectively;

whereas, the bulk density, particle density and total

porosity of the casting increased by 0.78±0.39,

2.66±0.19 and 0.60±0.18% respectively (Figures 1

and 2). As number of days progressed, there was a

steady decline in the moisture content, WHC, total

porosity and WFPS with all the treatments, and it

was more pronounced in castings stored in partially

sealed bags. In the partially sealed bags,

approximately 65% of moisture loss was recorded

during the 12 weeks of storage. The castings stored

in the airtight bags showed about 8% moisture loss

probably due to evaporation during sample

preparation.

During storage, the rapid change in the

moisture content of the castings stored in partially

sealed bags reduced the structural pores of organic

matter resulting in the increase in bulk density. In

this treatment, the bulk density increased more

sharply between the first to sixth week and in the

remaining period the changes was slow. The bulk

density of castings increased about twofold in

partially sealed bags. In this treatment, changes in

the bulk density of castings positively correlated

(p<0.001) with gravimetric water loss and WFPS in

the respective treatment (Table 1). The castings

stored in airtight sealed bags showed about 20%

increase in bulk density in the course of 12 weeks

of storage. Similarly, the particle density of castings

increased significantly in partially sealed bags than

the airtight bags. The particle density of castings

increased about 19 and 6% with partially sealed and

airtight bags, respectively. Changes in particle

density of castings depend on the degree of

decomposition, which reported to be varying

between 1.0 and 1.5 g cm-3

(Hassink, 1995;

Ruhlmann et al., 2006). During the storage, there

was no significant influence of pre-drying process

on particle density of the castings stored in both

partially sealed and airtight bags.

Total porosity of castings decreased around

20% in partially sealed bags, whereas it was less

than 6% with castings stored in airtight bags.

Nevertheless, the WFPS of castings showed a

different trend of results, which reduced maximally

in airtight sealed bags than castings of partially

sealed bags. The reduction in WFPS of castings

stored in airtight bags was 15 and 19% with un-

205

Figure 1. Changes in the moisture content (a), bulk density (b), particle density (c) and water holding capacity (d), of

un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2 respectively) and un-dried and pre-dried

castings stored in partially sealed bags (T3 and T4 respectively), at different periods of time.

dried and pre-dried castings respectively. In the

partially sealed bags, 10 and 12% reduction in

WFPS was observed with un-dried and pre-dried

castings. During storage, the castings of partially

sealed bags developed low WFPS which may be

due to loss of structural pores during dehydration

and decomposition. This was reported with peat by

Schwärzel et al. (2002) and Kechavarzi et al.

(2010). The changes in the WFPS of vermicast

have a distinct impact on the other physical,

chemical and biological properties; WFPS is of

primary importance in the regulation of

hydrological properties, gas diffusion, microbial

colonization, nutrient mineralization etc. (Gorres et

al., 2001; Schjønning et al., 2011).

In general, water in the castings is retained in

pore spaces and adsorbed onto the organic matter

(Chaudhuri et al., 2009). In this study, a significant

linear relationship (p<0.001) was found between

water holding capacity and porosity of castings

(Table 1). In the partially sealed bags, WHC of the

castings reduced around 60% with both un-dried

and pre-dried castings, whereas it was 9 and 16%

with respective to airtight bag treatments. It

indicates that the decrease in WHC of castings

during the ageing process was likely due to the

20

30

40

50

60

70

80

0 2 4 6 8 10 12

Mo

istu

re c

on

ten

t %

No. of weeks

a

0.25

0.35

0.45

0.55

0.65

0 2 4 6 8 10 12

Bu

lk d

en

sity

g c

m-3

No. of weeks

b T1 T2 T3 T4

1.2

1.3

1.4

1.5

1.6

0 2 4 6 8 10 12

Par

ticl

e d

en

sity

g c

m-3

No.of weeks

c

100

200

300

400

500

0 2 4 6 8 10 12

WH

C %

No.of weeks

d

206

Figure 2. Changes in the total porosity (a), water filled porosity (b), pH (c) and EC (d), of un-dried and pre-dried


sealed bags (T3 and T4 respectively), at different periods time. reduction of pore space in the castings. Moreover, a

decrease in hydrophilic components of organic

matter e.g. polysaccharides (Piccolo and Mbagwu,

1999) during the storage might be restricting the

available surface area that absorbs water, resulting

in the decline in water holding capacity of castings

(Li et al., 2007).

The castings stored in both airtight and

partially sealed bags showed little reduction in the

pH throughout the experimental period. The

decrease in pH during the storage may be due to

CO2 and organic acid generated by the microbes

during the decomposition of organic matter (Elvira

et al., 1998). Moreover, the mineralization of

nitrogen and phosphorous into nitrites/nitrates and

orthophoshates and bioconversion of organic

material into intermediate species of organic acids

may also have contributed to lowering the pH of

vermicast (Ndegwa and Thompson, 2000; Suthar,

2007). The EC of castings stored in both airtight

and partially sealed bags showed increasing trend

and it was up to twofold higher than the fresh

castings.


There was a significant influence (p<0.001)

55

60

65

70

75

80

0 2 4 6 8 10 12

`To

tal P

oro

sity

%

No. of weeks

a

20

24

28

32

36

0 2 4 6 8 10 12

WFP

S %

No. of weeks

b T1 T2 T3 T4

6.8

7.0

7.2

7.4

7.6

7.8

0 2 4 6 8 10 12

pH

No.of weeks

c

1

2

3

4

5

0 2 4 6 8 10 12

EC m

mm

ho

s cm

-1

No.of weeks

d

207

Figure 3. Changes in the total organic carbon (a), dissolved organic carbon (b), total nitrogen (c), ammonium nitrogen

(d), nitrate nitrogen (e) and available phosphorus (f), of un-dried and pre-dried castings stored in airtight sealed bags

(T1 and T2 respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at

different periods of time.

300

320

340

360

0 2 4 6 8 10 12

Org

anic

Car

bo

n m

g g-1

No. of weeks

a

0

2

4

6

0 2 4 6 8 10 12

DO

C m

g g-1

No. of weeks

b T1 T2

T3 T4

10

20

30

40

0 2 4 6 8 10 12

Tota

l nit

roge

n m

g g-1

No. of weeks

c

0

200

400

600

0 2 4 6 8 10 12

Am

mo

niu

m µ

g g-1

No. of weeks

d

0

200

400

600

0 2 4 6 8 10 12

Nit

rate

µg

g-1

No.of weeks

e

0

100

200

300

0 2 4 6 8 10 12

Ph

osp

ho

rus

µg

g-1

No.of weeks

f

208

on chemical properties of vermicast with pre-drying

and storage (Table 2). Pre-drying of vermicast

showed reduction in organic carbon, DOC, total

nitrogen, NH4-N, extractable form of phosphorus,

potassium and calcium content of vermicast

(Figures 3 and 4). The NO3-N and sulfate content of

castings increased by 51.2±2.8, 13.5±1.0%,

respectively. The castings stored in the partially

sealed bags showed higher reduction of C content

than airtight bags and it was about 12% during the

12 weeks of storage; whereas, it was less than 3%

in castings stored in airtight bags. The higher

reduction of C content in partially sealed bags was

due to the rapid mineralization of C by aerobic

microbes (van Gestel et al., 1993) and the reduction

was maximum in the first two weeks. Few studies

on the ageing of castings of anecics and endogeics

have also reported high C reduction in the initial

period (Martin, 1991; Burtelow et al., 1998;

Scullion et al., 2003). The reduction in C

mineralization as the days progressed might be the

reason for reduction in WFPS which holds a greater

proportion of bacteria (Hassink et al., 1993). The

reduction in this pore size during the storage

curtails the microbial access to the substrate

(Gorres et al., 2001).

In the partially sealed bags, more than 86%

of DOC was exhausted in the 12 weeks storage. In

the case of castings stored in the airtight bags there

was increasing trend in DOC content during second

to fourth week and then there was a slow decline

till the end. However, the reduction rate in this

treatment was about half comparing to the partially

sealed bags. Less reduction of DOC in the airtight

bags may be due to the low C utilizing efficiency of

anaerobic microbial community (Søndergaard and

Middelboe, 1995; Song et al., 2008). The extent of

sealing of vermicast packs also showed significant

differential response to N loss from the vermicast.

The high aeration in the partially sealed bags would

have promoted the volatilization of ammonium

present in the fresh vermicast (Ndegwa and

Thompson, 2000; van der Stelt et al., 2007), hence,

there was around 25% loss of nitrogen during the

first two weeks of storage and the total N loss was

46% in 120 days.

The result of the present study is in contrast

to the report of ageing studies on vermicast in

which the total N of castings was rather constant

during the entire ageing process (Decaëns et al.,

1999; Jiménez and Decaëns, 2004). Studies

reported by Decaëns et al. (1999) and Jiménez and

Decaëns (2004) were conducted with castings of

size of up to 6 cm diameter and an average dry

weight of 25 g produced by the large anecic

earthworm, Martiodrilus sp. The larger size of the

castings may impede aeration and prevent the

ammonium from volatilization within the

aggregates, whereas castings stored in the present

study was < 2mm dia. Moreover, in the above cited

studies the pH of castings might be around 5 as the

pH of the study area was 4.5 and 5. This inference

is drawn based on the previous reports on Amynthas

khami, another anecic earthworm. The pH of fresh

castings was 4.3 to 5.7 where the pH of bulk soil

was in the range of 4.0 to 5.3 (Jouquet et al., 2008).

Generally, in different animal manure, large

ammonia volatilization takes place between a pH of

7 and 10: below pH 7 ammonia volatilization

decreases, and completely stops at pH of about 5

(Hartung and Phillips, 1994). Therefore, lower pH

of these castings may be the reason for no N loss

during the ageing. In addition, the diet of the above

mentioned anecic earthworm species consisted of a

considerable portion of mineral soil and the

castings they produced had a lesser concentration of

ammonium (220 to 290 µg g-1

) which was around

threefold lower than the castings used in the present

study.

The concentration of NH4+

-N in the fresh

castings was 525.6±8.2 mg kg-1

. The high NH4+

content in the fresh castings may be due to

mineralized nitrogen from the substrate during the

gut passage and the addition of urine in the

posterior part of the digestive tract (Decaëns et al.,

1999). The castings stored in partially

209


during the storage.

Treatment Moisture

content

Bulk

density

Particle

density

Water holding


Water filled

pore space EC pH


0.257***

450.8***

484.3***

19.02***

57.91***

34.50***

459.3***


11951.1***

9426.5***

1811.2***

5920.0***

195.6***

13889.2***

78.37***


Treatment 32.13

*** 23.63

*** 516.5

*** 110.9

*** 0.935

n.s 5.31

n.s 814.2

*** 73.76

***

*p<0.05,

**p<0.01,

***p<0.001,



storage

Treatment

Total

organic

carbon

Dissolved

organic

carbon

Total

nitrogen

Ammonium-

nitrogen

Nitrate-

nitrogen

Available

phosphorus

Exchangeable

potassium

Available

sulfur

Exchangeable

calcium

Exchangeable

sodium


86.57***

18.56***

7118.8***

191.4***

792.1***

36.44***

7679.8***

1014.7***

1165.1***

Pre-Treatment 313.0**

14198.0***

593.1**

26891.3***

23530.2***

1293.7***

10243.5***

660.1***

9912.2***

13756.7***

Extend of Sealing

X Pre-Treatment 13.94

** 10.35

* 12.49

** 442.4

*** 23.58

** 1.309

n.s 190.7

*** 2971.0

*** 374.2

*** 1758.1

***

*p<0.05,

**p<0.01,

***p<0.001,



storage


activity

Cellulase

activity

β-Glucosidase


Alkaline

phosphatase

activity

Arylsulphatase

activity

Microbial

biomass carbon


*** 703.6

*** 63.82

*** 903.1

*** 2311.4

*** 92.91

*** 18..15

**

Pre-Treatment 316645.8

*** 65.80

*** 11661.0

*** 1261.1

*** 76.24

*** 5681.6

*** 397.6

***


Treatment 2048.5***

1785.9***

1545.3***

3.706 n.s

3956.0***

7.259* 36.80

***

*p<0.05,

**p<0.01,

***p<0.001,


210

Figure 4. Changes in the extractable potassium (a), sulfur (b), calcium (c) and sodium (d) of un-dried and pre-dried



sealed bag showed a constant reduction in the NH4+

concentration and it was reflected by an increase in

NO3 concentration due to the nitrification of

existing ammonium pools (Lavelle and Martin,

1992; McInerney and Bolger, 2000). In this

treatment, about 21% of reduction in ammonium

content was observed in the first week, and then

there was more than 75% reduction at the end of 84

days. The results are agreeing with the findings on

N dynamics of castings generated from the

endogeic species Pontoscolex corethrurus,

Aporrectodea caliginosa and the epigeic Metaphire

hilgendorfi during the ageing process (Lavelle et

al., 1992; Aira et al., 2005; Kawaguchi et al.,

2011). In these studies, most of the ammonium

present in the vermicast was exhausted rapidly as

recorded in the present study. The castings stored in

airtight bags showed maximum NH4+ reduction of

18% during the first week of storage and then there

was a slow reduction till the end. The anoxic

condition created in airtight bags after a week of

storage would have impeded the nitrification

process (Jouquet et al., 2011). However, there was

slow reduction in NO3- observed throughout the

storage process along with gradual reduction in

NH4+, certainly as the result of denitrification and

microbial immobilization (Lavelle and Martin,

1992; Decaëns et al., 1999).

15

20

25

30

35

0 2 4 6 8 10 12

Po

tass

ium

µg

g-1

No. of weeks

a

0

200

400

600

800

0 2 4 6 8 10 12

Sulf

ate

µg

g-1

No. of weeks

b T1 T2

T3 T4

80

120

160

200

240

280

0 2 4 6 8 10 12

Cal

ciu

m µ

g g-1

No.of weeks

c

3.0

5.0

7.0

9.0

0 2 4 6 8 10 12

Sod

ium

µg

g-1

No.of weeks

d

211

Figure 5. Changes in the dehydrogenase (a), β- glucosidase (b), cellulase (c), urease (d), alkaline phosphatase (e) and

arylsulphatase (f) enzymes activity of un-dried and pre-dried castings stored in airtight sealed bags (T1 and T2

respectively) and un-dried and pre-dried castings stored in partially sealed bags (T3 and T4 respectively), at different

periods of time.

0

20

40

60

80

100

0 2 4 6 8 10 12

De

nh

ydro

gen

ase

µg

INT

g-1 2

h-1

No. of weeks

a

0

20

40

60

80

100

0 2 4 6 8 10 12

β-G

luco

sid

ase

act

ivit

y µ

g P

NG

g-1

h-1

No. of weeks

b T1 T2

T3 T4

0

1

2

3

4

0 2 4 6 8 10 12

Ce

llula

se a

ctiv

ty µ

g C

MC

g-1

24

h-1

No. of weeks

c

0

40

80

120

160

200

0 2 4 6 8 10 12

Ure

ase

act

ivty

µg

NH

4-N

g-1

2 h

-1

No. of weeks

d

0

200

400

600

800

0 2 4 6 8 10 12

Ph

osp

hat

ase

act

ivit

y µ

g P

NP

g-1

h-1

No.of weeks

e

0

20

40

60

80

0 2 4 6 8 10 12

Ary

lsu

lph

atas

e a

ctiv

ity

µg

PN

S g-1

h-1

No.of weeks

f

212

The available phosphorus content of the

castings stored in both airtight and partially sealed

bags increased by 20% in the un-dried castings, and

about 10% in the pre-dried castings during the first

week of storage, and then it progressively declined

over the 84 days of storage. In the initial weeks of

storage, largely available nutrient pool and high

moisture content enhanced phosphatase activity; the

resultant higher mineralization increased the

fraction of labile P pool in the vermicast

(Parthasarathi and Ranganathan, 1999; Flegel and

Schrader, 2000). The findings are very similar to

the reports on castings of Lampito mauritii,

Martiodrilus carimaguensis and other species

(Satchell and Martin, 1984; Parthasarathi and

Ranganathan, 1999; Jimenez et al., 2003). Further

storage, decreased the inorganic P content of

castings of partially sealed bags and the reason may

be due to reduction in microbial activity which in

turn is due to developing low moisture content.

Although similar trend of labile P content was

observed in the case of airtight bags, the amount of

labile P was higher than the partially sealed bags.

The reason may be due higher unutilized available

P pool due to reduction condition in this treatment

(Reddy et al., 2011).

During storage, the exchangeable K content

declined in all the treatments. The decreased

exchangeable K content in partially sealed bags was

about four times higher than the casting of airtight

bags. The amount of reduction in exchangeable K

was 12 and 46% with partially sealed and airtight

bags respectively. The variation in the response of

exchangeable K during storage with different

treatments could not be attributed to any particular

reason as relevant reports on dynamics of K in the

vermicast and even on the biogeochemistry of K in

soils are very limited. The sulfate content of the

vermicast stored in the partially sealed bags

increased by 25% during the first week of storage

and then there was a decline till the end. Increase in

arylsulphatase activity in this treatment indicates

the increase in microbial mineralization of sulphur

during this period. Further storage decreased the

sulfate content of vermicast and the reason may be

attributed to high C/S ratio of above 600 during this

period. The high C/S ratio might have promoted

the immobilization of available sulfur (Tabatabai

and Chae, 1991; Reddy et al., 2002). In the case of

airtight bags, except in the first week of storage, the

inorganic S was always higher than the castings of

partially sealed bags.

The castings stored in the airtight bags

showed fluctuation in both Ca and Na content

during the entire period of storage. In this

treatment, the maximum of 47% increase in Ca

content was observed during the tenth week of

storage with pre-dried castings. The Na content of

the pre-dried castings stored in the air tight bags

showed a maximum increase of 33% at the second

week followed by 17% with un-dried castings of

the same week. In the case of partially sealed bags,

both Ca and Na content steadily declined till the

end of experimental period. The increase in Ca and

Na content in the airtight bags may be attributed to

increased solubility of organic carbon, and

increased competition between the cations for the

negatively charged sites due to increased levels of

Fe and Mn under reducing conditions (Wolt, 1994;

Phillips and Greenway, 1998).


The changes in enzyme activity of vermicast

during the pre-drying and storage with different

treatments are presented in Figure 5. The β-

glucosidase, urease, alkaline phosphatase and

arylsulphatase activities of vermicast decreased by

14.6±0.4, 22.4±1.6, 11.3±4.9 and 14.8±2.9%

respectively during pre-drying. Whereas, the

dehydrogenase and cellulase activity increased by

1.77±2.63 and 29.1±2.6%, respectively. In the

airtight bags, the dehydrogenase activity increased

threefold with un-dried and pre-dried castings at

fourth and seventh week, respectively. After this,

there was a steady decline in dehydrogenase

activity till the end of the experiment. The rapid

exhaustion of oxygen present in the airtight bags

213

would have been bringing a shift of the activity

from aerobic to anaerobic microorganisms which

could be the reason for the higher dehydrogenase

activity (Brzezinâska et al., 1998). Many studies on

soil enzyme activity reported similar high

dehydrogenase activities in anoxic condition

(Glinski et al., 1986; Stepniewska et al., 1990;

Brzezinâska et al., 1998). The castings stored in the

partially sealed bags, showed a 19% increase in

dehydrogenase activity during the first week of

storage. After that there was a steady decline in

enzyme activity till the end of the experimental

period and it was about sixfold lower than the fresh

castings.

Unlike dehydrogenase, the β-glucosidase and

arylsulphatase activities of vermicast were higher in

the partially sealed bags than airtight bags. A

maximum of 74 and 77% increase in enzyme

activity was observed in un-dried and pre-dried

castings stored in partially sealed bags during the

third week of storage; whereas, it was 53 and 70%

in corresponding airtight bag treatments. At the end

of the experimental period, half of the initial

enzyme activity was reduced in both partially

sealed and airtight bags. The arylsulphatase activity

in partially sealed bags increased by 15 and 31%

with un-dried and pre-dried castings, respectively

during the initial period of storage, and then there

was a steady decline by 97% lower than the fresh

casting. The castings stored in airtight bags showed

steady decline in enzyme activity from the initial

day of storage and the maximum reduction of 97%

in the 12 weeks. The decline in arylsulphatase

activity in airtight bags is probably due to the

development of anoxic condition in this treatment

(Kang and Freeman, 1999; Xiao-Chang and Qin,

2006).

The cellulase, urease and alkaline

phosphatase activities of vermicast showed varying

response to different treatments. The cellulase

activity of pre-dried castings stored in airtight bags

and un-dried castings of partially sealed bags

increased by twofold during the first three weeks of

storage. Whereas, un-dried castings of the airtight

bags and pre-dried castings of partially sealed bags

showed 6 and 16% increase in cellulase activity

during the first week of the storage. The results

indicate that the castings which was exposed to

aeration either for short period during the pre-

drying or continuously in partially sealed bags has

increased the cellulase activity. The reduction in the

cellulase activity during further storage may be due

to the reduction in available inorganic N as reported

on soil and plant litter (Sinsabaugh et al., 2005;

Yao et al., 2009). The alkaline phosphatase activity

of vermicast increased by twofold in un-dried

castings of airtight bags and more than threefold

with others during the first week of storage. A

possible explanation for the high phosphatase

activity during the pre-drying and first week of

storage may be due to high microbial activity in

vermicast during this period. Increase in labile N

and other nutrient content in the fresh vermicast

probably increased the P demand, a likely

consequence of higher phosphatase activity

stimulated by the active microbial population

(Allison et al., 2007). The increase in labile P

content by stimulated high enzyme activity may

have contributed to the slow decline in phosphatase

activity in the continuing weeks of storage. A

maximum urease activity was observed in the fresh

castings probably due to a considerable portion of

urea present in the castings as earthworm urine

(Edwards, 2004). During the storage, the declining

trend of urease activity was seen in all the

treatments till the end of the experimental period.

The decline in urease activity during the storage

may be due to the reduction in urea content and

denaturing of enzyme (Gould et al., 1973). In this

experiment, about 6 and 15 times reduction in

urease activity was recorded at the end of 12 weeks

of storage.

Microbial biomass carbon in the fresh

vermicast increased by 12% during pre-drying. The

storage of vermicast greatly affected the microbial

biomass C content of castings stored in both airtight

and partially sealed bags (Figure 6). During the first

214



airtight sealed bags (T1 and T2 respectively) and un-

dried and pre-dried castings stored in partially sealed

bags (T3 and T4 respectively), at different periods of

time.

week of storage, increase in the microbial biomass

C was observed in all the treatments, and the

increase was predominant in the castings stored in

partially sealed bags. Although other studies

showed reduction (Parthasarathi and Ranganathan,

1999; Tiunov and Scheu, 2000; Scullion et al.,

2003) or no changes (Aira et al., 2005, 2010) in

biomass during in the initial period of ageing, in the

present study, higher microbial activity and

biomass was observed during the first week. Most

of these previous studies are field based and

conducted with anecic and endogeic earthworm

species, which are reported to have a considerable

portion of mineral particles in their castings and

organic matter (Blanchart et al., 2004; Jouquet et

al., 2008) than the castings of epigeic species,

which is subject to exhaustion very soon by intense

microbial activity (Tiunov and Scheu, 2000; Aira et

al., 2005).

In the present study, castings generated from

the phytomass attributed abundant nutrient

availability in the fresh vermicast, which would

have created hotspot of microbial activity during

the first week of storage and increased their

biomass. Continuing storage of castings was

characterized by reduction in microbial C in the

course of 12 weeks storage. The progressive

decline of available nutrient and moisture content

of vermicast was related to the resultant decline in

microbial biomass in further storage (Scheu, 1987).

The higher reduction of microbial C in the airtight

treatment could be due to the elimination of aerobic

microbial groups and shift in microbial community

due to induced anoxic condition. Microbial

communities under anoxic condition has a low

energy yield from metabolizing reduced substrates,

leading to low C use efficiency. An unpublished

data reported by Song et al. (2008) shows that

microbial biomass C accounted for 2.28, 1.79, and

0.99% of soil organic C in oxic, intermittent, and

anoxic soils, respectively. The microbial

community structure with increasing oxygen

demand lead to the emergence of other stress

factors. For example, the nutritional stress indicator

in the anoxic soils was two to fourfold of that in the

oxic soils (Song et al., 2008).

4. Conclusions

The physical, chemical and biochemical

properties of vermicast were significantly altered

during the storage with different methods used in

this study. Amongst the treatment, the castings

stored in the partially sealed bags showed drastic

reduction in porosity, WHC and most of the plant

available nutrient studied. The bulk density of

castings also increased maximally in the castings

stored in partially sealed bags. These changes may

impede the water availability, oxygen diffusion and

plant root penetration and also low nutrient

availability in the vermicast when applied in field.

In addition, the excess dryness of castings stored in

partially sealed bags would have highly stabilized

the nutrients in organic matter and this would

impede the instant nutrient release to plants in the

field. The changes in the microbial and enzyme

activities of the castings support all the assumptions

mentioned above. Although, the beneficial

properties reduced in the castings stored in airtight

bags, it was less intense than the partially sealed

0

5

10

15

20

25

0 2 4 6 8 10 12

Bio

mas

s ca

rbo

n m

g g-1

No.of weeks

T1 T2 T3 T4

215

bags. Since the physical properties of these castings

also favor the growth of microbes even after

prolonged storage, the disappeared nutrient pool

can be regained once it is applied in the field. Even

though, pre-drying reduced some of the beneficial

properties of vermicast, this practice can be

recommended before storage as it prevents the

disintegration of the structure of castings due to

excess moisture; and would enhance the physical

properties and microbial activity.

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enzymatic activity in a loess soil (Horizon Ap).

Folia Soc. Sci. Lublinensis 30, 53-59.

Suthar, S., 2007. Production of vermifertilizer from

guar gum industrial wastes by using

composting earthworm Perionyx sansibaricus

(Perrier). Environmentalist 27, 329-335.


Arylsulfatase activity of soil. Soil Sci. Soc.

Am. Proc. 34, 225-229.



479-487.

Tabatabai, M.A., Chae, Y.M., 1991. Mineralization

of sulfur in soils amended with organic wastes.

J. Environ. Qual. 20, 684-690.





van der Stelt, B., Temminghoff, E.J.M., van Vliet,

P.C.J., van Riemsdijk, W.H., 2007.

Volatilization of ammonia from manure as

affected by manure additives, temperature and

mixing. Bioresour. Technol. 98, 3449-3455.

van Gestel, M., Merckx, R., Vlassak, K., 1993.

Microbial biomass responses to soil drying and

rewetting: The fate of fast- and slow growing

microorganisms in soils from different

climates. Soil Biol. Biochem. 25, 109-123.



agriculture. John Wiley and Sons, New York,

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waterlogged and aerobic incubation on enzyme

activities in paddy soil. Pedosphere 16, 532-

539.






Biochem. 41, 1425-1432.

SUMMARY AND CONCLUSION

Chapter

14

219

CChhaapptteerr 1144

Summary and conclusion

Vermicomposting is the name given to the

process of conversion of biodegradable matter by

earthworms into vermicast. In general, vermicast

has been found to improve the seed germination,

growth, and productivity of many plant species. It

is also seen to improve soil health in terms of

aeration, water holding, and pH neutralization.

Although, there is accumulating scientific evidence

on these aspects, our understanding is largely

confined to the influence of vermicast derived from

the manure-based or manure-augmented substrates

on soils and plants whereas studies on vermicast

generated solely from phytomass are lacking. It has

also been a matter of concern whether phytomass-

based vermicompost will be as beneficial to the soil

and plants as manure-based vermicompost is. This

concern is particularly relevant for plants like

lantana and ipomoea which are known to possess

constituents that are toxic to animals and other

species of plants.

Information on the food preference and the

ageing of castings has been well documented for

geophagous and geophytophagous earthworms.

Much less attention has been paid towards

phytophagous earthworms. As vermicomposting is

primarily carried out using phytophagous species, it

is important to cover this knowledge gap as well.

The first chapter of the thesis comprises of

brief introduction to the studies reported in

Chapters 3 to 14. The importance of the studies

carried out by us in the context of the prior art is

brought out in Chapter 2.

Chapters 3-5 are dedicated to understanding

the feeding behavior of the epigeic earthworms,

which are species extensively used for

vermicomposting of various types of organic waste.

This group of earthworms is phytophagous in habit

and is believed to prefer organic matter, principally

phytomass. In contrast, the anecics and endogeics

consume an appreciable amount of mineral soil

with organic matter which facilitates assimilation of

nutrients in earthworm gut probably by keeping

their gizzard muscles toned up. To check whether

epigeics will ingest soil/sand even when there is

luxury availability of phytomass, set of experiments

were carried out with epigeic species, Eudrilus

eugeniae, and neem, ipomoea and cow dung as

feed.

It was seen that even though initially E.

eugeniae did ingest sand and soil despite the luxury

availability of phytomass, this tendency reduced as

the time passed indicating adaptive response to the

phytomass feed. Moreover, the assimilation of

soil/sand particles in the vermicast increased the

bulk density and particle density, reduced the pore

space, water holding capacity and nutrient content

of castings, which may reduce their beneficial

impact on plant growth and soil. These findings are

of significance in vermireactor’s design and

optimization because they indicate that sand-soil-

gravel bedding as used in conventional

vermireactors is necessary neither to ensure the

survival, growth and fecundity of the earthworm

nor the quality of the vermicompost.

Chapters 6-9 describe the effect of

vermicompost derived from the allelopathic weeds

such as lantana (Lantana camara) and ipomoea

(Ipomoea carnea), and paper waste on the

germination, growth and fruition of cluster bean

(Cyamopsis tetragonoloba). The impact of the

vermicompost (VC) was compared with that of an

220

inorganic fertilizer (IF) which had all the main

macro and micro-nutrients in concentrations

equivalent to the ones present in the VC. It was

seen that significantly greater germination rates

occurred in the lantana based VC treatments

compared to controls, while ipomoea based VC

showed suppression, indicating the presence of

some of the germination inhibitory components in

vermicast. However, both lantana and ipomoea

based VC also supported better plant growth in

terms of stem diameter, shoot length, shoot mass,

number of leaves, and leaf pigments. The positive

impact extended up to pod yield. In addition,

vermicast application enhanced root nodule

formation, reduced disease incidence, and limited

stunted plants.

The findings reveal that the allelopathic

ingredients of these weeds seem to have been

totally eliminated during the course of its

vermicomposting, and that these vermicompost has

the potential to support growth and fruit yield better

than equivalent quantity of inorganic fertilizers.

These studies open up the possibility that other

allelopathic weeds, and also plants which are toxic

in other ways, may be utilizable as substrates in

high-rate vermireactors as vermicomposting is

likely to destroy the toxic components of these

substrates as it is seen to have done in case of

lantana and ipomoea. This, in turn, may

enormously enhance the applicability of

vermicomposting as well as provide a means of

utilizing the biomass of several invasives which,

otherwise, goes to waste. In case of paper waste

based VC, application had no beneficial impact on

growth of cluster bean plant; whereas, the

corresponding IF treatments exhibited better seed

germination and plant growth than the former one.

In this experiment, the plants did not fructify in

both VC and IF treatments. The low fertile

experimental soil and insufficient minor and trace

nutrient present in the VC generated from the paper

waste might have impeded the production of

vegetables.

The findings of the studies reported in

Chapters 6-8 revealed that the vermicompost

derived from different parent materials have

different physical, chemical and biological

qualities, and their impact on germination, growth

and yield of plant also varied considerably.

Therefore, to understand the impact of

vermicompost from different parent materials on

soil health which is directly related to the growth

and yield of plants was assessed and reported in

Chapter 9. Several sets of experiments were carried

out in which the vermicast generated from different

organic wastes such as paper waste, leaves of

ipomoea, and of lantana or inorganic fertilizers

were applied in potting soil anchoring cluster bean.

Samples from all these treatments were collected on

weekly basis during different stages of plant

growth. The results reveal that vermicast

application created a conducive physical

environment by reducing bulk density, and

improving the water holding capacity and porosity

of soil. In addition, throughout the experiment, the

nutrient content of the soil was significantly higher

compared to inorganic fertilizers treated one.

Although, the inorganic fertilizers application

initially increased the nutrient content in soil, as the

days progressed, substantial quantity of applied

nutrient became unavailable to the plants. The loss

probably may be due to high leaching of mineral

nutrients from the soil. Apparently, no significant

impact has been observed on physical properties of

soil with inorganic fertilizers application.

Consequently, the vermicompost amendment may

be considered a good strategy for improving plant

growth, which reduces the deterioration of

agricultural lands due to the rampant use of

inorganic fertilizers.

The studies reported in Chapters 10-13,

which are perhaps the first of the kind, explores the

changes in the physical, chemical and biological

properties of vermicast that occur during storage

with the objective of finding conditions that

minimize the deterioration in the fertilizer value of

the vermicast. Vermicast generated from different

221

organic waste such as paper waste, neem and cow

dung was packed in airtight and partially sealed

bags with and without pre-drying for 24 hours.

Changes in several physical, chemical, and

biological properties of the castings were monitored

for three months with weekly assessments. The

manner of storage was seen to influence the plant-

friendly attributes of vermicast in a strong fashion.

Airtight storage after pre-drying was the most

beneficial, followed by airtight storage of the fresh,

undried, vermicast. In partially sealed storage there

was significantly more rapid deterioration of the

beneficial attributes than in airtight storage.

Interestingly, whereas 24-hr pre-drying before

airtight storage was helpful in retaining the plant-

friendly attributes of the vermicast for longer than

fresh-airtight storage, pre-drying before partially

sealed storage had the opposite effect. Apparently,

partially sealed storage added to the water loss that

had already occurred during the pre-drying, and

brought the water content below a level that was

needed to support biological activity within the

vermicast matrix. This indicates that a certain level

of water content is most appropriate for retaining

the microbiological and enzyme activities of the

vermicast; and the presence of water above or

below that level hastens the cast’s ageing. Further

work should be aimed at determining the most

beneficial water levels and how best to retain them.

In the present chapter, the gist of all the

studies reported in this thesis is given in brief.

STANDARDIZATION OF ANALYTICAL METHODS

Appendix

223

AAppppeennddiixx

Standardization of analytical methods

There are a number of analytical methods available for the variables assessed in the present study but in soil

or plant matrices. It is not known whether the methods will be applicable to vermicast also. Therefore, some

of the available analytical procedures for a few selected variables were identified on the basis of required

precision and accuracy. The procedures were then assessed for their applicability to vermicompost matrix by

means of the standard addition method, in which varying concentration of standards were spiked with

samples, and analyzed from its quantitative recovery. After ascertaining near-quantitative recovery with

adequate precision the methods were short-listed and utilized. The following table gives the details of the

standard addition methods for selected variables utilized for the experimental works detailed in the previous

chapters.

Parameters Methods Standards

Amount of

standard

added

Recovery

percentage

(Mean±SD) References

Total organic

carbon

Modified dichromate

redox method Glucose anhydrate 10-100 mg g

-1 94.5±2.4 Heanes, 1984

i

Total nitrogen Modified Kjeldahl method Ammonium nitrate 10-100 mg g-1

96.5±3.9 Kandeler,

1993ii

Ammonium

Extraction: Potassium

chloride extraction

Determination: Modified

indophenol blue method/

Nitroprusside catalyst

method

Ammonium sulfate 0.1-10 mg g-1

97.1±3.9 Bashour and

Sayegh, 2007iii

Nitrate

Extraction: Potassium

chloride extraction

Determination: Devarda’s

alloy method

Potassium nitrate 0.1-10 mg g-1

94.5±2.4

Gavlak et al.,

1994iv; Jones,

2001v

Extractable

Phosphorus

Extraction: Mehlich 3

extraction

Determination:

Ammonium molybdate-

ascorbic acid method

Potassium

dihydrogen

phosphate

0.1-10 mg g-1

96.8±3.6

Knudsen and

Beegle, 1988vi;

Mehlich,

1984vii

Extractable

Sulphate

Extraction: Calcium

chloride extraction

Determination: Turbidi-

metric method

Potassium sulfate 0.1-10 mg g-1

95.7±4.1

Houba et al.,

2000viii

;

Bashour and

Sayegh, 2007iii

Exchangeable

Potassium Extraction: Neutral

ammonium acetate

solution extraction

Determination: Flame

photometry

Potassium chloride 0.1-10 mg g-1

95.5±3.6

Lavkulich,

1981ix

Exchangeable

Calcium Calcium carbonate 0.1-10 mg g

-1 97.0±2.2

Exchangeable

Sodium Sodium chloride 0.1-10 mg g

-1 96.8±2.4

224

For each parameter, the Shewhart charts/control charts were prepared to assess the bias and precision

of standardization data (Figures 1-9). For that, the samples were fortified with known concentration of

chemical of interest, and quantitative recovery was calculated using following formula.

The quantitative recovery was plotted over the cumulative mean of the repeated measures. Control lines were

drawn which represent mean + 2 sigma (upper and lower warning limits, UWL and LWL) and mean + 3

sigma (upper and lower control limits, UCL and LCL). In these graphs, the warning lines (UWL and LWL)

represent a 95% confidence interval, and the control lines were corresponding to a 99% confidence interval

for acceptability of the methods for the specific matrix. In general, a single value outside the control lines is

considered unacceptable. The control charts for the few selected variables are shown in Figures 1-9. The

precision level and consistence in results for these variables are in acceptable range, hence these methods

have been utilized for analyzing the constituents in vermicast matrix.

Figure 1. Control chart of the quantitative recovery of organic carbon in vermicast matrix; where UCL – upper control

level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning level,

mean – 2σ; LCL – lower control level, mean – 3σ.

Figure 2. Control chart of the quantitative recovery of total nitrogen in vermicast matrix, where UCL – upper control



86

89

92

95

98

101

104

Sp

ike

reco

ver

y %

82

88

94

100

106

112

Sp

ike

reco

ver

y %

UWL

Mean

LWL

LCL

UCL

UWL

Mean

LWL

LCL

UCL

225

Figure 3. Control chart of the quantitative recovery of ammonium in vermicast matrix, where UCL – upper control



Figure 4. Control chart of the quantitative recovery of nitrate in vermicast matrix, where UCL – upper control level,

mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning level, mean –

2σ; LCL – lower control level, mean – 3σ.

Figure 5. Control chart of the quantitative recovery of extractable phosphorus in vermicast matrix, where UCL – upper

control level, mean + 3σ; UWL – upper warning level, mean + 2σ; mean – average of values; LWL – lower warning

level, mean – 2σ; LCL – lower control level, mean – 3σ.

83

88

93

98

103

108 S

pik

e re

cov

ery

%

86

90

94

98

102

106

Sp

ike

reco

ver

y %

83

88

93

98

103

108

Sp

ike

reco

ver

y %

UWL

Mean

LWL

LCL

UCL

UWL

Mean

LWL

LCL

UCL

UWL

Mean

LWL

LCL

UCL

226

Figure 6. Control chart of the quantitative recovery of extractable sulfur in vermicast matrix, where UCL – upper



Figure 7. Control chart of the quantitative recovery of exchangeable potassium in vermicast matrix, where UCL – upper



Figure 8. Control chart of the quantitative recovery of exchangeable calcium in vermicast matrix, where UCL – upper



83

88

93

98

103

108

Sp

ike

reco

ver

y %

83

88

93

98

103

108

Sp

ike

reco

ver

y %

89

92

95

98

101

104

Sp

ike

reco

ver

y %

UWL

Mean

LWL

LCL

UCL

UWL

Mean

LWL

LCL

UCL

UWL

Mean

LWL

LCL

UCL

227

Figure 9. Control chart of the quantitative recovery of exchangeable sodium in vermicast matrix, where UCL – upper



i Heanes, D.L., 1984. Determination of total organic-C in soils by an improved chromic acid digestion and

spectrophotometric procedure. Commun. Soil Sci. Plant Anal. 15, 1191-1213. ii Kandeler, E., 1993. Bestimung von Gesamtstickstoff nach kjeldahl. In: Schinner, F., Kandeler, E., Ohlinger, R.,

Margesin, R. (eds.), Bodenbiologische Arbeitsmethoden, pp. 346-366. Spinger, Berlin. iii

Bashour, I., Sayegh, H.A., 2007. Methods of analysis for soils of arid and semi-arid regions. Food and Agriculture

Organization of the United Nations, Rome. iv Gavlak, R.G., Horneck, D.A., Miller, R.O., 1994. Plant, Soils, and Water Reference Methods for the Western Region,

Western Regional Extension Publication WREP 125, University of Alaska, Fairbanks, 45–47. v Jones, J.B., 2001. Laboratory guide for conducting soil tests and plant analysis. CRC Press, New York.

vi Knudsen, D., Beegle, D., 1988. Recommended phosphorus tests. In: Dahnke, W.C. (eds.), Recommended chemical

soil tests procedures for the North Central region, p. 12-15. Bulletin No. 499 (Revised), North Dakota Agric. Exp. Sta.,

Fargo, North Dakota. vii

Mehlich, A., 1984. Mehlich 3 soil test extractant: A modification of the Mehlich 2 extractant. Commun. Soil Sci.

Plant Anal. 15, 1409-1416. viii

Houba, V.J.G., Temminghoff, E.J.M., Gaikhorst, G.A., van Vark, W., 2000. Soil analysis procedures using 0.01 M

calcium chloride as extraction reagent. Commun. Soil Sci. Plant Anal. 31, 1299–1396. ix

Lavkulich, L.M. 1981. Methods Manual, Pedology Laboratory. University of British Columbia, Vancouver, British

Columbia, Canada.

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UWL

Mean

LWL

LCL

UCL

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