universiti putra malaysia upmpsasir.upm.edu.my/id/eprint/70415/1/fp 2017 37 ir.pdf · 2019. 7....
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UNIVERSITI PUTRA MALAYSIA
In-Situ IMMOBILIZATION OF SELECTED HEAVY METALS USING DIFFERENT SOURCE OF PHOSPHATE AMENDMENTS FOR ORGANIC
PRODUCTION OF MISAI KUCING (Orthosiphon stamineus Benth)
KHAIRUN NAIM BIN MULANA@MAULANA
FP 2017 37
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In-Situ IMMOBILIZATION OF SELECTED HEAVY METALS USING
DIFFERENT SOURCE OF PHOSPHATE AMENDMENTS FOR ORGANIC
PRODUCTION OF MISAI KUCING (Orthosiphon stamineus Benth)
By
KHAIRUN NAIM BIN MULANA@MAULANA
Thesis Submitted to the School of Graduate Studies, Universiti Putra Malaysia,
in Fulfilment of the Requirements for the Degree of Master of Science
January 2017
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All materials contained within the thesis, including without limitation text, logos,
icons, photographs and all other artwork, is copyright material of Universiti Putra
Malaysia unless otherwise stated. Use may be made of any material contained within
the thesis for non-commercial purposes from the copyright holder. Commercial use of
material may only be made with the express, prior, written permission of Universiti
Putra Malaysia.
Copyright © Universiti Putra Malaysia
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Abstract of thesis presented to the Senate of Universiti Putra Malaysia in fulfilment
of the requirement for the degree of Master of Science
In-Situ IMMOBILIZATION OF SELECTED HEAVY METALS USING
DIFFERENT SOURCE OF PHOSPHATE AMENDMENTS FOR ORGANIC
PRODUCTION OF MISAI KUCING (Orthosiphon stamineus Benth)
By
KHAIRUN NAIM BIN MULANA@MAULANA
January 2017
Chairman : Professor Che Fauziah bt. Ishak, PhD
Faculty : Agriculture
Sustainable organic herbal production requires utilization of bio-waste materials as
plant nutrient sources due to its economical aspect and would ensure continuous
productivity. The usage of organic wastes in organic farming system (OFS) as
fertilizers and soil amendments should be monitored as the accumulation of heavy
metals has been reported in several studies. This study was set up to investigate the
effect of chicken manure soil application on heavy metals concentration in organically
grown O. stamineus and a potential approach to immobilize it using phosphate based
amendments. Glasshouse trials has been conducted for study 1 and study 3 whilst
incubation study was conducted for study 2 accordingly. Results in the study 1
indicated that the trace metals of lead (Pb) and Zn (Zn) exceeds the threshold value
allowed by Pharmacopeia in herbs and by Malaysian Food Act (1983) and Food
Regulations (1985) at 10 and 40 mg/kg respectively. This is postulated due to
application of chicken manure and organic materials during cultivation. In study 2, 2
sources of apatite were tested: - 1) Egyptian rock phosphate (ERP) and 2) bone meal
(BM) under incubation study. The results demonstrated that the dominant lead forms
were mainly associated with residual, carbonates and also exist in exchangeable form.
From this study, the suitable rate for reducing phyto-available lead in the soil system
applied with 10 t/ha chicken manure is at 1 t/ha of Egyptian rock phosphate and 2 t/ha
of bone meal. In the third study, the effectiveness of phosphorus on reducing heavy
metals were evaluated under controlled condition in the glasshouse. Results
demonstrated heavy metals accumulated in the aerial parts of O. stamineus with copper
(18.3-37.7 mg/kg), zinc (60.9-92.0 mg/kg) and lead (1.9-3.6 mg/kg). From the
fractionation study, the percentage of exchangeable fraction was reduced with
application of P-amendments with the highest of 20.2% reduction recorded for 2 t/ha
application of triple super phosphate. This is followed by reduction in exchangeable
fraction for others treatments: 2 t/ha of bone meal (4.1%), 4 t/ha of bone meal (5.1%),
1 t/ha of Egyptian rock phosphate (8.1%) and 2 t/ha of Egyptian rock phosphate at
17.6%. Applications of P-amendments tested shows effectiveness of TSP in reducing
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lead in soils. The best treatments are as follows: - 2 t/ha of triple super phosphate> 2
t/ha of Egyptian phosphate rock> 1 t/ha of Egyptian rock phosphate> 4 t/ha of bone
meal> 2 t/ha of bone meal. These treatments were recorded as being able to stabilize
the Pb as indicated in the percentage reduction of phytoavailable pools into a more
stable form of residual pool.
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Abstrak tesis yang dikemukakan kepada Senat Universiti Putra Malaysia sebagai
memenuhi keperluan untuk ijazah Master Sains
IMMOBILISASI LOGAM BERAT SECARA In-Situ DENGAN
MENGGUNAKAN SUMBER FOSFORUS BERBEZA BAGI PENGELUARAN
SECARA ORGANIK MISAI KUCING (Orthosiphon stamineus Benth)
Oleh
KHAIRUN NAIM BIN MULANA@MAULANA
Januari 2017
Pengerusi : Profesor Che Fauziah bt. Ishak, PhD
Fakulti : Pertanian
Pengeluaran herba secara lestari memerlukan penggunaan bahan bio-sisa sebagai
sumber nutrien terutamanya dalam sistem pertanian secara organik. Secara amnya,
penggunaan bahan sisa organik ini dalam pertanian organik sebagai baja atau peminda
tanah perlu dipantau kerana pengumpulan logam berat telah dilaporkan dalam
beberapa kajian terdahulu. Penyelidikan ini dijalankan bagi mengkaji kesan
penggunakan tahi ayam terhadap pengambilan logam berat oleh O. stamineus dan
pendekatan yang sesuai dalam mengawalnya menggunakan peminda berasaskan
fosfat. Ujian rumah kaca telah dijalankan bagi kajian 1 dan kajian 3, manakala kajian
pengeraman dijalankan bagi kajian 2. Keputusan dalam kajian pertama menunjukkan
bahawa kandungan logam berat plumbum melepasi had yang dibenarkan oleh
Pharmacoepia pada kadar 10 mg/kg bagi herba dan tumbuhan ubatan. Manakala,
kandungan logam berat zink telah melepasi had yang dibenarkan oleh Akta Makanan
Malaysia (1983) dan Peraturan-Peraturan Makanan (1985) pada kadar 40 mg/kg. Ini
berkemungkinan disebabkan oleh penggunaan tahi ayam dan bahan-bahan organik
semasa penanaman. Dalam kajian 2, 2 sumber apatite diuji: - 1) batuan fosfat dari
Mesir (ERP) dan 2) serbuk sisa tulang (BM) dalam kajian pengeraman. Keputusan
menunjukkan bahawa pecahan fraksi yang dominan berkait-rapat dengan fraksi sisa
baki, karbonat dan dalam bentuk bertukar ganti. Daripada kajian ini, kadar yang sesuai
untuk mengurangkan plumbum pada sistem tanah menggunakan 10 t/ha baja tahi ayam
iaitu 1 t/ha batuan fosfat dari Mesir dan 2 t/ha sisa tulang. Dalam kajian yang ketiga,
keberkesanan fosforus mengurangkan logam berat telah dinilai di dalam keadaan
terkawal rumah kaca. Keputusan menunjukkan logam berat terkumpul di bahagian tisu
O. stamineus adalah seperti berikut: tembaga (18.3-37.7 mg/kg), zink (60.9-92.0
mg/kg) dan plumbum (1.9-3.6 mg/kg). Daripada kajian pemeringkatan, peratusan
pecahan ditukar-ganti telah dikurangkan dengan penggunaan peminda fosforus dengan
yang tertinggi adalah pengurangan 20.2% yang dicatatkan pada aplikasi 2 t/ha triple
super fosfat. Ini diikuti dengan pengurangan dalam pecahan ditukar-ganti pada
rawatan lain: 2 t/ha sisa tulang terlerai (4.1%), 4 t/ha sisa tulang terlerai (5.1%), 1 t/ha
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batuan fosfat dari Mesir (8.1%) dan 2 t/ha batuan fosfat dari Mesir pada kadar 17.6%.
Penggunaan peminda fosforus diuji menunjukkan keberkesanan TSP dalam
mengurangkan plumbum dalam tanah. Rawatan terbaik adalah seperti berikut: - 2 t/ha
triple super fosfat> 2 t/ha batuan fosfat dari Mesir> 1 t/ha batuan fosfat dari Mesir> 4
t/ha sisa tulang terlerai> 2 t/ha sisa tulang terlerai. Rawatan ini dapat menstabilkan Pb
seperti yang ditunjukkan dalam pengurangan peratusan fraksi yang boleh diambil
tumbuhan ke dalam bentuk yang lebih stabil, fraksi sisa baki.
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ACKNOWLEDGEMENTS
In the name of Allah, the Most Gracious and the Most Almighty
Alhamdulillah, all praise to almighty Allah for the strength and His blessing in
completing this thesis. Special appreciation goes to my supervisor, Prof. Dr. Che
Fauziah Ishak, for her invaluable guidance, constructive comment and constant
support throughout the experimental work and conduct of thesis. My appreciation also
goes to Prof. Dr. Rosenani Abu Bakar for the support and guidance given.
I would like to express my gratitude to all the laboratory staff and technicians at the
Department of Land Management, Faculty of Agriculture, Universiti Putra Malaysia
(UPM).
Last but not least, I would like to thank Majlis Amanah Rakyat (MARA), for the
financial support given which makes my study possible.
I wish to express my thanks to my parents, Maulana Abdul Shukor and Kamariah Daud
as well as my siblings for their support given throughout this study. Without their
encouragement and understanding it would have been impossible for me to finish this
study. Thank you so much.
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This thesis was submitted to the Senate of Universiti Putra Malaysia and has been
accepted as fulfilment of the requirement for the degree of Master of Science. The
members of the Supervisory Committee were as follows:
Che Fauziah Ishak, PhD
Professor
Faculty of Agriculture
Universiti Putra Malaysia
(Chairman)
Rosenani Abu Bakar, PhD
Professor
Faculty of Agriculture
Universiti Putra Malaysia
(Member)
ROBIAH BINTI YUNUS, PhD
Professor and Dean
School of Graduate Studies
Universiti Putra Malaysia
Date:
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Declaration by graduate student
I hereby confirm that:
this thesis is my original work;
quotations, illustrations and citations have been duly referenced;
this thesis has not been submitted previously or concurrently for any other degree
at any other institutions;
intellectual property from the thesis and copyright of thesis are fully-owned by
Universiti Putra Malaysia, as according to the Universiti Putra Malaysia
(Research) Rules 2012;
written permission must be obtained from supervisor and the office of Deputy
Vice-Chancellor (Research and Innovation) before thesis is published (in the form
of written, printed or in electronic form) including books, journals, modules,
proceedings, popular writings, seminar papers, manuscripts, posters, reports,
lecture notes, learning modules or any other materials as stated in the Universiti
Putra Malaysia (Research) Rules 2012;
there is no plagiarism or data falsification/fabrication in the thesis, and scholarly
integrity is upheld as according to the Universiti Putra Malaysia (Graduate
Studies) Rules 2003 (Revision 2012-2013) and the Universiti Putra Malaysia
(Research) Rules 2012. The thesis has undergone plagiarism detection software.
Signature: ________________________ Date: __________________
Name and Matric No.: Khairun Naim Bin Mulana@Maulana (GS39440)
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Declaration by Members of Supervisory Committee
This is to confirm that:
the research conducted and the writing of this thesis was under our supervision;
supervision responsibilities as stated in the Universiti Putra Malaysia (Graduate
Studies) Rules 2003 (Revision 2012-2013) are adhered to.
Signature:
Name of Chairman of
Supervisory Committee:
Che Fauziah Ishak, PhD
Signature:
Name of Member of
Supervisory Committee:
Rosenani Abu Bakar, PhD
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TABLE OF CONTENTS
Page
ABSTRACT i
ABSTRAK iii
ACKNOWLEDGEMENTS v
APPROVAL vi
DECLARATION viii
LIST OF TABLES xiii
LIST OF FIGURES xiv
LIST OF PLATES xv
LIST OF ABBREVIATIONS xvi
CHAPTER
1 INTRODUCTION 1
2 LITERATURE REVIEW 3
2.1 Herbs and the demands 3
2.1.1 Orthosiphon stamineus or ‘misai kucing’ 3
2.1.2 Soil suitability, climatic and nutrient requirement 4
2.1.3 Trace metals phytoaccumulating potential of O.
stamineus
5
2.2 Organic production system vs. conventional production system 5
2.2.1 Presence of heavy metals in Organic Farming System 6
2.2.2 Chicken manure as organic fertilizer 7
2.3 Heavy metals 7
2.3.1 Effects of heavy metals on human health 8
2.3.2 Factors affecting availability of trace metals in well-
drained soil
8
2.3.3 Soil of Malaysia and background study of trace metals 9
2.3.3.1 Bungor series 10
2.3.4 Maximum Permissible Limits (MPLs) 10
2.3.4.1 Lead essential or not? 11
2.3.4.2 Sources of lead 11
2.3.4.3 Movement of lead to plants 11
2.4 In-situ immobilization techniques 12
2.4.1 Apatite application to reduce lead 12
2.4.2 Mechanism of lead immobilization and other heavy
metals by phosphates
13
2.4.3 The stability and adsorption capacity of Pb-phosphate
formation
13
2.4.4 Cadmium presence in the phosphates fertilizers 14
2.5 Summary of Literature Review 14
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3 MATERIALS AND METHODS 15
3.1 Characterization of soil, organic materials and P amendments 15
3.1.1 Physical properties 15
3.1.2 Chemical properties 15
3.2 Analysis 17
3.2.1 Plant analysis 17
3.2.2 Soil analysis 18
4 RESULTS AND DISCUSSION 21
4.1 Trace metals accumulation, growth performance, nutrient uptake
and soil properties in organically grown Orthosiphon stamineus
as influenced by chicken manure and rice husk biochar.
21
4.1.1 Experimental design 21
4.1.2 Sample preparation 22
4.1.3 Statistical analysis 22
4.1.4 Characterization 22
4.1.5 Growth performances of O. stamineus 24
4.1.6 Nutrients uptake by O. stamineus 25
4.1.7 Soil properties 26
4.1.8 Trace metal concentrations in O. stamineus 27
4.1.9 Conclusion 30
4.2 In-situ immobilization of lead using phosphate rock and bone
meal-incubation study
30
4.2.1 Experimental design 30
4.2.2 Sample preparation 31
4.2.3 Statistical analysis 32
4.2.4 Characterization 32
4.2.5 Sources of lead 33
4.2.6 Egyptian rock phosphate 35
4.2.7 Bone meal 37
4.2.8 Conclusion 40
4.3 In-situ immobilization of lead using phosphate rock, bone meal
and triple super phosphate-glasshouse trial
40
4.3.1 Experimental design 40
4.3.2 Sample preparation 41
4.3.3 Transfer factor 41
4.3.4 Statistical analysis 42
4.3.5 Characterization 42
4.3.6 Growth performances of O. stamineus 43
4.3.7 Dry weight of O. stamineus 43
4.3.8 Nutrients uptake by O. stamineus 44
4.3.9 Soil properties 45
4.3.10 Trace metals concentrations of O. stamineus 45
4.3.10.1 Lead 46
4.3.10.2 Zinc 47
4.3.10.3 Copper 48
4.3.11 The O. stamineus phytoremediator abilities 49
4.3.12 Heavy metals concentration in soil 50
4.3.13 Lead phase association 50
4.3.14 Transfer factors of heavy metals 53
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4.3.15 Correlation between Pb uptakes and soil fractions 54
4.3.16 Conclusion 54
5 SUMMARY, CONCLUSION AND RECOMMENDATIONS FOR
FUTURE RESEARCH
55
REFERENCES
57
APPENDICES 63
BIODATA OF STUDENT 79
LIST OF PUBLICATIONS 80
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LIST OF TABLES
Table Page 2.1 9
2.2 10
2.3 10
3.1 19
4.1 23
4.2 24
4.3 25
4.4 27
4.5 28
4.6 32
4.7 36
4.8 37
4.9 38
4.10 39
4.11 42
4.12 43
4.13 44
4.14 45
4.15 46
4.16 50
4.17 53
4.18 53
4.19
The 95% ‘Investigation Levels’ determined for Malaysia (n=241
soils).
Maximum Permissible Limits based on Food Acts (1983) and Food
Regulation (1985).
Permissible limits of medicinal plants (WHO, 2013).
Extractants used for the fractionation study.
Selected chemical properties of soil and organic media.
Effect of different rates of biochar and chicken manure on biomass
of O. stamineus produce.
Effect of application rates of biochar and chicken manure on
selected nutrients concentration and uptake of O. stamineus.
Effect of different application rates of rice husk biochar and
chicken manure on soil properties.
Effect of different application rates of bochar chicken manure on
heavy metals concentration in O. stamineus aerial parts.
Selected chemical properties of soil, chicken manure and
phosphorus materials.
The effect of chicken manure(0, 5, 10 and 20 t/ha), Egyptian rock
phosphate (0, 1, 2 and 4 t/ha) on soil pH, available phosphorus,
exchangeable Pb, carbonates Pb, Fe & Mn oxides Pb, organic
matter Pb and residual Pb.
Pearson correlation coefficients table between available P and soil
pH against fractions of soil and total Pb for Egyptian phosphate rock
application (n=48).
The effect of chicken manure(0, 5, 10 and 20 t/ha), bone meal (0,
1, 2 and 4 t/ha) on soil pH, available phosphorus, exchangeable Pb,
carbonates Pb, Fe & Mn oxides Pb, organic matter Pb and residual
Pb.
Pearson correlation coefficients table between available P, soil pH
and soil fractions, total Pb for bone meal application (n=48).
Chemical properties of triple super phosphate
Effect of different P-amendments application on biomass of O.
stamineus produce.
Effect of P-amendments application on selected plant nutrients
concentration of O. stamineus.
Effect of P-amendments on selected soil properties of O.
stamineus.
Effect of different P-amendments application on heavy metals
contents in O. stamineus aerial plant tissues.
Effect of different application P-amendments on total heavy metals
concentration in soil.
Effect of P-amendments on fraction of Pb in the soil system.
Effect of different application P-amendments on transfer factor of
lead.
Pearson correlation coefficients between Pb uptakes and soil
fractions (n=24).
54
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LIST OF FIGURES
Figure Page 2.1 The projection of global market value of herbal industry. 5
4.1 Fractionation of control and soil applied with 5, 10 and 20
t/ha of chicken manure.
34
4.2 Lead concentration of plant tissue of O.stamineus when
applied with different P-amendments.
47
4.3 Zinc content of plant tissue of O.stamineus when applied
with different P-amendments.
48
4.4 Copper content of plant tissue of O.stamineus when applied
with application of P-amendments.
49
4.5 The lead as per percentage in 5 fractions; 1) exchangeable,
2) carbonates, 3) Fe & Mn oxides, 4) organic matter and 5)
residual with application of different P-amendments.
51
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LIST OF PLATES
Plate Page 2.1 The O.stamineus or Misai Kucing (Javanese Tea). 4
4.1 Growth performances observes in O.stamineus applied with
different rates of chicken manure (0, 2.5 and 5 t/ha) and
biochar 10 t/ha, combination of biochar 10 t/ha with 2.5 and
5 t/ha chicken manure.
25
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LIST OF ABBREVIATIONS/NOTATIONS/GLOSSARY OF TERMS
AA AutoAnalyzer
AAS Atomic Absorption Spectrometer
ANOVA Analysis of Variance
BM Bone Meal
CRD Completely Randomized Design
CRBD Completely Randomized Block Design
EC Electrical Conductivity
ERP Egyptian Rock Phosphate
DOA Department of Agriculture
MPL Maximum Permissible Limits
rpm Rotation per minute
TSP Triple Super Phosphate
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CHAPTER 1
INTRODUCTION
1.1 General background
Food safety and environmental concerns have gained attention among consumers
recently which results in increasing demands of organic farming produce over those
of conventional farming. Agronomic practices, such as application of soil amendment
and organic fertilizer (types and rates) for organic production of herbs on Malaysian
soils have not been well investigated. These practices should be reviewed due to
potential of heavy metals accumulation in the herbal produce. Therefore, safety
assurances of organic herbs products should be monitored along with the enhancement
of biomass yield. Chicken manure and organic fertilizers might contribute to higher
trace metals uptake in plant tissues particularly for lead. On the other hand, location of
farms close to the roadside can also result in higher Pb content of soil due to
automobiles emission using leaded gasoline. Through application of phosphorus-
amendments, such as phosphate rock and bone meal, Pb can be immobilized or fixed
in the soil system.
1.2 Justification & problem statement
Herbs production under organic farming system (OFS) claims to assure the safety of
organic produce compared to conventional farming system (CFS). However, OFS
heavy usage of organic amendments and organic fertilizers potentially contribute to
accumulation of heavy metals. Through fractionation studies the information on
reactivity and potential bioavailability of heavy metals could be obtained.
According to study 1, zinc (Zn) seemed to be the heavy metals that exceeded the
maximum permitted concentration (MPC) by Malaysian Food Act (1983) and Food
Regulations (1985) at 40 mg/kg. Similarly, lead (Pb) concentration surpassed the
Pharmacopeia threshold value (10 mg/kg) in the organically grown misai kucing (O.
stamineus). On the other hand, the copper (Cu) concentration can be a potential hazard
as the amount indicated is near the 30 mg/kg maximum permitted concentration (MPC)
by Malaysian Food Act (1983) and Food Regulations (1985). There are few reports
related to copper, zinc and lead accumulations in O. stamineus being published. Abdu
et al., (2011) concluded that O. stamineus can be a potential phytoremediator for Pb
and Zn, whilst Manan et al., (2015) indicated that O. stamineus acts as a moderate
accumulator for Zn, Cu and Pb. However, zinc and copper are essential for both plants
and human, which unlike lead, possess threat even at minimum amounts. Thus, in
order to reduce lead contents in the O. stamineus herbal produce, the application of P-
amendments such as phosphate rock or bone meal could be a potential option. The
formation of stable lead compounds and the process of immobilization through soil
fractionation study in phosphate-amended soil will be investigated in this study.
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For a sustainable herbal production, utilization of bio-waste materials as a nutrient
source in organic farming is economical and would ensure continuous productivity.
However, documented research on the effects of organic fertilizers towards safety and
quality of herbs produced are still lacking.
1.3 Objectives
The general objective of this study was to investigate the effect of chicken manure
application on heavy metals concentration in organically grown of O. stamineus and a
potential approach to immobilize it using phosphate-based amendments.
The specific objectives of the study were:
1) To assess heavy metals (Cu, Zn, Pb, As, Cd and Hg) uptake in organically grown
O. stamineus applied with different rates of chicken manure and biochar as soil
amendments.
2) To determine the optimum rate of phosphate rock and bone meal in order to
immobilize Pb in soil applied with high level of chicken manure based on
incubation study.
3) To assess the effectiveness of selected rates and types of P-amendments indicated
earlier to immobilize Pb in organically grown O. stamineus on soil applied with
high level of chicken manure. Secondary assessment on Cu and Zn concentrations
would be indicated accordingly.
1.4 Hypotheses
Organic farming system of herbs does not guarantee safety of the produce in terms of
heavy metals accumulation. The phytoavailablity of heavy metals in the soil system
can be confirmed through fractionation study. Lead contents in herbal produce can
efficiently be reduced through land application of P amendments such as phosphate
rock, bone meal and triple super phosphate to form a stable complex of lead phosphate
minerals.
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