journal of research in biology volume 3 issue 1

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Fernando Reboredo [Archaeobotany, Forestry, Ecophysiology] New University of Lisbon, Caparica, Portugal

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Table of Contents (Volume 3 - Issue 1)

Serial No Accession No Title of the article Page No

1 RA0304 Anatomical variation in the olfactory apparatus of marine teleosts.

Biswas S, Datta NC, Sarkar SK and De SK.

742-746

2 RA0306 The use of purple yam (Dioscorea trifida) as a health-promoting

ingredient in bread making.

Teixeira AP, Oliveira IMA, Lima ES and Matsuura T.

747-758

3

RA0305

Bioefficacy of Novaluron®, a chitin synthesis inhibitor against the

tropical warehouse moth, Ephestia cautella.

Sackey I, Eziah VY and Obeng-Ofori D.

759-767

4

RA0308

A Checklist of Butterflies of Meenachil River Basin, Kerala, India.

Vincy MV, Brilliant R and Pradeepkumar AP.

768-774

5 RA0325 Microbial production of glutaminase enzyme.

Mario Khalil Habeeb.

775-779

6 RA0307 A review on the role of nutrients in development and organization of

periphyton.

Saikia SK, Nandi S, Majumder S.

780-788

7 RA0187 Assessing heavy metal contamination of road side soil in urban area.

Sarala Thambavani D and Vidya Vathana M.

789-796

http://jresearchbiology.com/documents/RA0304.pdf







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Biology

Anatomical variation in the olfactory apparatus of marine teleosts

Keywords: Olfactory, Rosette, Scombridae, Carangidae, Platycephalidae, etc.

ABSTRACT: The olfactory apparatus of marine teleosts viz., Rastrelliger kanagurta, Scomberoides commersonianus and Platycephalus scaber belonging to the family of Scombridae, Carangidae and Platycephalidae respectively has been fixed in 10% formaldehyde solution for 24 h and anatomically examined under light microscope (LM). Anatomical variation regarding the nostrils, olfactory rosette, occurrence of accessory nasal sacs, olfactory lobes, length of the olfactory nerve tracts, etc. are observed. These morphological variations may denote species specific and may decisive for several biological functions.

742-748 | JRB | 2013 | Vol 3 | No 1

This Open Access article is governed by the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.

www.jresearchbiology.com

Journal of Research in Biology

An International Open Access

Research Journal

Authors:

Biswas S1, Datta NC2,

Sarkar SK1 and De SK1.

Institution:

1. Department of Zoology,

Vidyasagar University,

Midnapore (West) - 721102,

West Bengal, India.

2. 110/20 B. T. Road,

Kolkata - 700108, West

Bengal, India.

Corresponding author:

Subrata Kumar De.

Email:

[email protected]

Phone No:

+91 03222 275329

Web Address: http://www.jresearchbiology.com/

documents/RA0304pdf.

Dates: Received: 08 Nov 2012 Accepted: 20 Nov 2012 Published: 09 Jan 2013

Article Citation: Biswas S, Datta NC, Sarkar SK and De SK. Anatomical variation in the olfactory apparatus of marine teleosts. Journal of Research in Biology (2013) 3(1): 742-748

Journal of Research in Biology An International Open Access Research Journal

Original Research

INTRODUCTION

Olfaction is an important type of chemoreception

which is mediated through well developed, complex and

organized olfactory system in vertebrates (Freitag et al.,

1999). This system generally develops from the

specialized tissue, called olfactory placode (von Kupffer,

1894). In the early stage of development, placodes are

formed from the preplacodal ectoderm of the anterior

region of the embryo, medial to epidermis and lateral to

both neural crest and neural plate (Knouff, 1935).

The developmental process leading to the formation of

fish olfactory organ, is little diverse (Hansen and

Zielinski, 2005). The peripheral olfactory organ is the

first chemosensory organ to develop in fish preceding the

systems of solitary chemosensory cells (Kotrschal et al.,

1997) and taste (Hansen et al., 2003). Fish generally

possesses a paired olfactory organ located at the anterior

part of the head (Derivot, 1984; Hansen and Zeiske,

1998; Døving, 2003; De and Sarkar, 2009). The olfactory

chambers, olfactory rosette, accessory nasal sacs,

olfactory bulbs, olfactory nerve tracts and brain are the

major components of fish olfactory apparatus

(Hamdani and Døving, 2007). The anatomy of the

olfactory apparatus shows wide range of structural

diversity regarding the shape and size of the olfactory

rosette, number of the olfactory lamellae, occurrence of

accessory nasal sacs, etc. among the teleostean groups

(Kleerekoper, 1969). The anatomical details of the

olfactory apparatus in several species belonging to the

diverse teleostean taxa with respect to their habitat are

still an obscure part in sensory biology. The sensory

systems of fishes show notable adaptations according to

habitat and mode of life in comparison with the higher

vertebrates (Bone and Moore, 2008). The present study

focused on the true anatomy of the olfactory apparatus in

three species of different ecological habitat viz.,

Rastrelliger kanagurta (Cuvier, 1816), a marine

oceanodromous fish; Scomberoides commersonianus

(Lacepede, 1801), a brackish water amphidromous fish

and Platycephalus scaber (Linnaeus, 1758), an estuarine

amphidromous fish to unfold their structural components

and functional significance in olfaction.

MATERIALS AND METHODS

Adult, sex-independent specimens of

R. kanagurta, S. commersonianus and P. scaber were

collected from the coastal regions of Bengal, India.

Specimens were preserved in 10% formaldehyde

solution for 24 h and brought to the laboratory. The

olfactory apparatus of these species were dissected out

and separately mount on grease free slide by using

glycerine. The olfactory apparatus of respective species

was examined under light microscope (LM).

RESULTS

R. kanagurta (Figure 1A) possesses two pairs of

nostril viz., anterior nostril and posterior nostril

(Figure 1B). The anterior nostril is an oval shaped

structure, encircled by a thick lip like ridge of skin and

located at the dorsal region of the snout where as the

posterior nostril is situated in front of the eye.

R. kanagurta shows a slit like structure i.e., posterior

nostril which is located at a moderate distance from the

anterior one (Figure 1B). The olfactory apparatus of the

said species is present at the antero-dorsal side of the

snout in between the anterior and posterior nostril

(Figure 1C). The multilamellar olfactory rosette is oval

in shape and is situated at the floor of the olfactory

chamber (Figures 1D and 1E). The triangular olfactory

lamellae vary from 60 to 70 in number per rosette and

are arranged pinnately on the raphe (Figure 1E). The

accessory nasal sacs are clearly marked in the olfactory

apparatus of R. kanagurta. The lacrimal sac is conical in

shape and located at the antero-medial region of

lachrymal bone in association with olfactory rosette

(Figures 1C and 1D). The ethmoidal sac is connected at

the posterior part of the olfactory rosette and situated

within a groove at the antero-lateral extension of the

Biswas et al., 2013

743 Journal of Research in Biology (2013) 3(1): 742-748

frontal bone (Figures 1C and 1D). The olfactory nerve

tracts are arised from the base of olfactory rosette and the

length varies from 16 mm to 18 mm respectively. The

distal part of the olfactory nerve tracts are connected

with olfactory lobes of the brain. The size of the

olfactory lobes is comparatively small in R. kanagurta

(Figure 1C).

In S. commersonianus (Figure 2A), the nostrils

are closely associated. The anterior nostril is an oval

shaped aperture and partly guarded by a nasal flap where

as the posterior nostril is crescentric in shape

(Figure 2B). The olfactory rosette is circular in shape and

consisting of two pairs of olfactory lamellae

(Figures 2C and 2D). The number of the olfactory

lamellae ranges from 100 to 140 per rosette (Figure 2D).

The accessory nasal sacs are not distinct in the olfactory

apparatus. The olfactory nerve tracts are arised from the

base of the olfactory rosette and the length is ranges from

10 mm to 12 mm. The olfactory lobe is comparatively

large in size (Figure 2C).

Interestingly, the nostrils of P. scaber

(Figure 3A) are situated at the antero-dorsal region to the

Biswas et al., 2013

Journal of Research in Biology (2013) 3(1): 742-748 744

Figure 1A - Schematic representation of Rastrelliger kanagurta.

Figure1B - The diagram shows oval shaped anterior nostril and transverse slit like posterior nostrils of

R. kanagurta.

Figure 1C - The olfactory apparatus of R. kanagurta shows olfactory rosette (OLR), lacrimal sac (LS),

ethmoidal sac (ES), olfactory nerve (OLN), olfactory lobe (OL), cerebral hemisphere (CHS),

optic lobe (OPL), cerebellum (CBL), medulla oblongata (MO), etc.

Figure 1D - The olfactory rosette (OLR) along with conical accessory nasal sacs viz., lacrimal sac (LS) and

ethmoidal sac (ETS).

Figure 1E - The dorsal view of oval shaped olfactory rosette shows central raphe (r), two rows of

lamella (l). The olfactory lamella is triangular in shape with prominent dorsal end (de), proximal end (pe)

and ventral margin (vm).

Plate - 1

OLR

LS

ETS

eye and lying far apart from each other. The anterior

nostril is oval in shape and has a tongue like nasal flap

where as the posterior nostril is valvular (Figure 3B).

The olfactory rosette is relatively large in size and oval

in shape (Figures 3C and 3D). The number of the

olfactory lamellae varies from 50 to 76 in number per

rosette (Figure 3D). The absence of accessory nasal sacs

is noted in the olfactory apparatus of P. scaber. The

length of the olfactory nerve tracts ranges from

22 mm - 24 mm. and it is well connected with the

olfactory lobe of the brain (Figure 3C).

DISCUSSION

Olfactory systems of fish are among the most

highly developed olfactory senses of vertebrates

(Kleerekoper, 1969). The sense of olfaction is mediated

through olfactory apparatus associated with nostrils. The

nostrils are responsible for incurrent and excurrent of

water during water ventilation (Nevitt, 1991). The

structure of the anterior and posterior nostril varies

among the teleostean fishes (Kapoor and Ojha, 1972).

The presence of nasal flaps in between the both nostrils

is almost common when they are closely associated

(Teichmann, 1954). The anterior and posterior nostril

probably acts as an avenue for water ventilation through

the olfactory rosette (Cox, 2008). The multilamellar

Biswas et al., 2013


Figure 2A - Schematic representation of Scomberoides commersonianus.

Figure 2B - The diagram shows anterior and posterior nostrils of S. commersonianus. Nasal flap (FL) is

distinct.

Figure 2C - The olfactory apparatus of S. commersonianus is comprises of olfactory rosette (OLR),

olfactory nerve (OLN), olfactory lobe (OL), cerebral hemisphere (CHS), optic lobe (OPL),

cerebellum (CBL), medulla oblongata (MO), etc.

Figure 2D - The circular olfactory rosette shows central raphe (r), two rows of lamella (l). The olfactory

lamella is large, triangular in shape with prominent dorsal margin (dm), dorsal end (de),

ventral margin (vm) and lingual process (lp).

Plate - 2

olfactory rosette perhaps adopt several type of

arrangement pattern (Holl, 1965). This lamellar

arrangement may help in the particular sensitivity to

certain components like amino acids, steroids,

prostaglandins, etc. (Theisen et al., 1991). Anatomically

the lamellar surface in S. commersonianus is much closer

due to the short distance between anterior and posterior

nostril than R. kanagurta and P. scaber. Therefore, the

water soluble odorants may travel short distance to

interact with comparatively greater olfactory lamellar

surface of S. commersonianus. The water ventilation is

assisted by the pumping mechanism of accessory nasal

sacs (Theisen et al., 1991) and may provide the ability

to sniff (Nevitt, 1991; Cox, 2008). R. kanagurta

possesses well developed accessory nasal sacs but

S. commersonianus and P. scaber has no accessory nasal

sacs. The movement of jaws and its associated muscles

are also very significant for the water ventilation

(Nevitt, 1991). Accessory nasal sacs may be found in the

olfactory organs of fishes with widely variable life styles

and habitats, both marine and fresh water which are not

confined to one particular situation (Cox, 2008).

The olfactory nerve may convey the chemical cues

during water ventilation to the brain (Hamdani and

Døving, 2007). The olfactory information plays an

important role in different behaviour of fish such as

Biswas et al., 2013


Plate - 3

Figure 3A - Schematic representation of Platycephalus scaber.

Figure 3B - The diagram shows anterior and posterior nostrils of P. scaber situated at a distance from each

other. Long nasal flap (FL) is also marked.

Figure 3C - The olfactory apparatus of P. scaber is comprised of olfactory rosette (OLR), olfactory nerve

(OLN), olfactory lobe (OL), cerebral hemisphere (CHS), optic lobe (OPL), cerebellum (CBL), medulla

oblongata (MO), etc.

Figure 3D - The circular olfactory rosette indicates central raphe (r) and two rows of lamella (l).

The olfactory lamella is triangular in shape with prominent dorsal margin (dm), dorsal end (de) and

proximal end (pe).

searching of foods, avoidance of predators,

discrimination between individuals of the same and

different, parental care, orientation in migration, etc.

(Hara, 1971). The olfactory system of teleosts is highly a

specialized structure for the recognition of various water

soluble chemical cues, so this may serve as a biological

model to monitor environmental health as well

as specific meagerness of the pollutants. The

xenotoxification of ocean especially the acidification

may also impair the ability of olfactory discrimination of

coastal and marine species (Munday et al., 2009). Thus,

it is necessary to examine the effect of specific toxic

agent at a subcellular level of olfactory structures in

marine teleolsts.

CONCLUSION

This comparative anatomical study on the

olfactory apparatus along with brain in three different

marine teleost belonging to the diverse ecological habitat

shows much structural variation according to the

changing environment which may be significant for

ecomorphology and evolutionary aspects of

neurobiology (Kotrschal et al., 1998). However, the

olfactory system of marine, estuarine and coastal or

migratory species may experience rapid fluctuations of

environmental inorganic ions (Hubbard et al., 2000), so

it could be an interesting part to identify the cellular

components that are involved in the ion regulation of the

olfactory apparatus in these migratory teleosts.

ACKNOWLEDGEMENTS

Authors are thankful to the Head, Department of

Zoology, Vidyasagar University, West Bengal, for

providing the necessary laboratory facilities.

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Ökologie der Tiere, 43(2):171-212.

Theisen B, Zeiske E, Silver WL, Marui T, Caprio J.

1991. Morphological and physiological studies on the

olfactory organ of the striped eel catfish, Plotosus

lineatus. Marine Biology, 110(1):127-135.

Von Kupffer C. 1894. Studien zur vergleichenden

Entwicklungsgeschichte des Kopfes der Kranioten. 2.

Heft . Die Entwicklung des Kopfes von

Ammocoetes planeri. Lehmann, Munich.

Biswas et al., 2013


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Biology

The use of purple yam (Dioscorea trifida) as a health-promoting

ingredient in bread making

Keywords: Purple yam (Dioscorea trifida); antioxidant activity; health-promoting food; Amazon region.

ABSTRACT: The use of purple yam (Dioscorea trifida) was evaluated as possible health-promoting ingredient in bread making in the state of Amazonas, Brazil. The centesimal composition, energy, and antioxidant activity of purple yam and its incorporated bread formulations (0%, 10%, 15% and 20%) were determined. An acceptance test and microbiological analysis of the formulations 10%, 15% and 20% were also performed. Except for lipids, the centesimal composition and caloric values revealed no statistically significant differences. An addition of purple yam in natura up to 20%, instead of wheat flour in ordinary bread (0%), can be made with no effect on the diet’s energy. The free radical scavenging, 2.2-diphenyl-1-picryl-hydrazyl (DPPH) and lipid per oxidation (LPO) methods revealed that the greater the percentage of purple yam being added into the breads the higher the antioxidant activity detected. The acceptance test applied to compare the three formulations of purple yam breads revealed a significant difference only in the attribute colour. Purple yam breads showed no preferable differences. Results highlight the feasibility of purple yam bread as a health-promoting food in the Amazon region.

747-758 | JRB | 2013 | Vol 3 | No 1

This Open Access article is governed by the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.




Research Journal

Authors:

Teixeira AP1,

Oliveira IMA1, Lima ES1

and Matsuura T2.

Institution:

1. Faculdade de Ciências

Farmacêuticas (FCF),

Universidade Federal do

Amazonas (UFAM), Rua

Alexandre Amorim, 330,

Aparecida, CEP: 69010-330,

Manaus, AM, Brasil.

2. Instituto de Ciências

Biológicas (ICB),

Universidade Federal do

Amazonas (UFAM), Av.

General Rodrigo Octávio

Jordão Ramos, 3000,

Campus Universitário,

Coroado I CEP: 69077-000,

Manaus, AM, Brasil.


Antonia Paiva Teixeira.

Email:

[email protected]

Web Address: http://www.jresearchbiology.com

document/ RA0306.pdf.


Article Citation: Teixeira AP, Oliveira IMA, Lima ES and Matsuura T.

The use of purple yam (Dioscorea trifida) as a health-promoting ingredient in bread making. Journal of Research in Biology (2013) 3(1): 747-758.


Original Research

mailto:[email protected]

INTRODUCTION

Yams belong to the family Dioscoreaceae, genus

Dioscorea (Pedralli, 1988; 1997; Pedralli et al., 2002;

Pedralli, 2004). This family is made up by 6 to 9 genera

comprising over 600 species distributed throughout the

World’s tropical, subtropical and temperate regions

(Barroso et al., 1974; Pedralli, 1988; 1997; Melo

Filho et al., 2000; Pedralli et al., 2002; Pedralli, 2004).

The yams (Dioscorea spp.) yield tubers, which are very

important as staple, nutritional and healthy food, and are

still used as an ingredient in traditional Chinese herbal

medicine. They show a worldwide distribution, and are

found in many tropical countries, in South-Eastern Asia

and Western Africa, where the species were introduced

by cultivators (Rasper and Coursey, 1967; Akanbi et al.,

1996; Omonigho and Ikenebomeh, 2000; Lin et al.,

2005). They can also be found in some American

countries, particularly in Brazil, where one can find them

in all regions, from the Amazon down to the Southern

part of the country (Chu and Figueiredo-Ribeiro, 1991;

Pedralli, 1997; 2004).

Purple yam (Dioscorea trifida) is an American

native species, which was domesticated by Amerindians,

with the cultivar distribution possibly pointing out its

domestication in Brazilian and Guyana border areas,

followed by dissemination throughout the Caribbean

islands (Pedralli, 1988; Pedralli et al., 2002; Pedralli,

2004). D. trifida shows a wide distribution in Central and

South America, from the Caribbean to Peru. In Brazil it

is found all the way from the Amazon right down to the

Southern region. The species is associated to forest

environments-Amazonian highland tropical rainforests,

Coastal Atlantic Forest in Southeastern Brazil and,

mesophytic (seasonable) and gallery forests (Pedralli,

1997).

Here in the Amazonian region, purple yam

(D. trifida) may be consumed in the following ways:

baked, boiled, mashed, as ingredients for soups and meat

stews, and in the formulation of flour for making cakes,

pies and porridges. Nevertheless, this species has

undergone little scientific investigation, so little is known

about its management techniques, genetic improvement,

nutritional potential, industrial use, storage procedures,

characterization, uses as natural dye, as well as its use as

a health-promoting ingredient, among others.

By and large, the bread consumed throughout the

world is made mostly of wheat flour, salt and yeast.

Many other ingredients, have been incorporated into

bread formulation, so as to increase its diversity and

product appeals (Hsu et al., 2004).

A few studies have highlighted the great

potential of purple yam in bread making. In this case,

yam flour may replace part of the wheat flour, improving

bread quality, as well as adding economical advantages

to it (Abramo, 1990; Hurtado et al., 1997; Litvin et al.,

1998; Omonigho and Ikenebomeh, 2000; Ratti, 2001).

Hsu et al., (2004) demonstrated the presence

of antioxidants in the flour of purple yam

(Dioscorea purpurea), in five formulations of breads

prepared with this tuber’s flour, with excellent

acceptance in Taiwan supermarkets. Contado et al.,

(2009) showed yam (Dioscorea spp.) mucilage-based

loaf to present good public acceptance as to flavor,

aroma and texture with sensory attributes, demonstrating

the use of this tuber to be feasible as improvers in bread

making.

The following aspects motivated the use of

purple yam (Dioscorea trifida) in natura as a bread

manufacturing health-promoting ingredient, in the

present work: 1) its significant world consumption,

presenting a considerable, expanding tillage alternative

(Rasper and Coursey, 1967; Abramo, 1990; IITA, 2007);

2) although, as yet incipient, an increase on the

production of this tuber in the State of Amazonas, Brazil,

especially in Caapiranga and Careiro Castanho

municipalities is being observed. According to the

Instituto de Desenvolvimento Agropecuário do Estado

do Amazonas (IDAM) in 2008, 110 families of the

Teixeira et al., 2013


Caapiranga municipality yielded 2,475 tonnes in an area

of 165 ha; and 3) the presence of antioxidants in purple

yam, which increases the nutritional capacity in breads

made from this tuber (Hsu et al., 2004).

The main aim of the present study was to

evaluate the potential of purple yam yield in the State of

Amazonas, Brazil as a health-promoting ingredient in

bread making. On this context, it determined the

centesimal composition, caloric value, and antioxidant

properties of purple yam as well as of breads made from

this tuber in natura. Then, it undertook an organoleptic

characteristic assessment of the breads, following tasters’

panel acceptance criteria. This purple yam species is, for

the very first time, being used in the Amazonian region,

as a feasible alternative for bread making.


Species identification and purple yam tuber

(Dioscorea trifida) collection

Identification of the species Dioscorea trifida

was accomplished by comparisons with a voucher

herbarium specimen (Exsicata number 1353) deposited

at the National Research Institute of Amazonia (INPA)

Herbarium. It is very common to find the purple yam

(D. trifida) exhibiting several color hues of its flesh

(edible portion), in Amazonas State Townships. The

types most easily identified are: roxinho (light purple

flesh); roxo (mid purple flesh); roxão (dark purple flesh);

branco (white flesh); and misto (white-purple flesh)

(Figure 1).

Purple yam samples were collected at two

Amazonas Townships: Caapiranga and Careiro

Castanho. Due to the seasonality and availability of these

tubers in the region, the centesimal composition analyses

of yams and breads were performed with Caapiranga

samples. Yam and bread antioxidant and bread sensory

and microbiological analyses were carried out with

Careiro Castanho samples.

Purple yam bread elaboration

On account of the probability of getting breads

with higher antioxidant concentration (Hsu et al., 2004),

roxão (dark purple flesh) type samples were used in the

present study (Figure 1C). Yams in natura, for replacing

wheat flour, were washed, peeled, weighed, ground

in the liquidizer together with yeast, oil and water.

Then, this mixture was added to the previously mixed

dry ingredients (wheat flour, powdered milk, sugar and

salt). Bread manufacturing formulations can be seen at

(Table 1). Homogenization (30 min.), dough underwent

initial fermentation (60 min.), intermediate time for

Teixeira et al ., 2013


B A

C D E

Figure 1. Flesh color varieties of the kinds of purple yam (Dioscorea trifida)

commonly found in fairs and markets of Manaus-AM. A) roxinho (light-purple

flesh); B) roxo (mid-purple flesh); C) roxão (dark purple flesh);

D) branco (white flesh); and E) misto (white-purple flesh).

bread shaping (25 min.), final fermentation (60 min.),

time for baking (30 min.) are followed for making yam

bread. After being prepared the breads were cooled to

room temperature and packed in polyethylene bags

displaying the product’s labeling.

Centesimal composition analyses of purple yam and

its incorporated breads

Centesimal composition analyses of purple yam

(Dioscorea trifida), and purple yam incorporated breads

in four formulations: 0%, 10%, 15% and 20%, were done

in triplicate. Moisture, ashes, lipid, proteins and crude

fiber contents were determined according to procedures

described by the Instituto Adolfo Lutz-IAL (2008).

Carbohydrate and caloric values were determined

according to the method of (AOAC, 2005).

Findings obtained on the bread formulation

centesimal composition analyses were subjected to



Table 1. Purple yam (D. trifida) incorporated

bread formulations.

Ingredients Type of bread*

0% 10% 15% 20%

Wheat flour (g) 500 450 425 400

Purple yam (g) 0 50 75 100

Sugar (g) 10 10 10 10

Salt (g) 5 5 5 5

Yeast (g) 10 10 10 10

Milk powder (g) 10 10 10 10

Oil (g) 10 10 10 10

Water (mL) 250 250 250 250

Total (g) † 545 545 545 545

*Percentage of wheat flour replaced by purple yam. †Total amount of ingredients used for preparing the

breads.

Parameters Yam

(D. spp.)* Yam

(D. alata)** Purple Yam (D.trifida)***

Moisture (%) 72.60 73.70 76.43 ± 0.50

Lipid (%) 0.20 0.10 1.13 ± 0.69 Protein (%) 2.00 2.30 1.83 ± 0.13

Crude Fiber (%)

0.60 7.30 1.80 ± 0.05

Ash (%) 0.90 0.90 0.78 ± 0.02 Carbohydrate

(%) 24.30 23.00 18.04 ± 0.66

Caloric value (Kcal/100g)

100.00 96.00 89.64 ± 4.52

Table 2. Yam centesimal composition.

*(Montaldo, 1977), **(TACO 2006), ***Present study Bre

ad

M

ois

ture

(%

) L

ipid

(%

) P

rote

in (

%)

Cru

de

Fib

er

(%)

Ash

(%

) C

arb

oh

yd

rate

(%

) C

alo

ric

va

lue

(kca

l/100 g

)

0%

29.7

9 ±

2.0

4a

4.4

9 ±

0.1

2ab

11.6

2 ±

0.8

1a

1.9

5 ±

0.0

8a

1.1

8 ±

0.2

8a

50.9

5 ±

2.7

9a

290.7

3 ±

7.5

6a

10%

31.0

3 ±

0.8

3a

4.2

4 ±

0,0

4a

10.6

7 ±

0,4

1a

1.9

1 ±

0,1

0a

1.4

1 ±

0.0

3a

53.4

1 ±

4.0

7a

294.4

5 ±

15.6

3 a

15%

32.6

5 ±

1.3

0a

4.8

7 ±

0.3

7ab

9.8

2 ±

1,2

4a

1.8

4 ±

0.0

7a

1.3

8 ±

0.0

7a

49.4

5 ±

2.2

6a

280.6

4 ±

3.9

5a

20%

35.0

9 ±

5.6

2a

4.7

4 ±

0.1

0b

10.0

6 ±

2,1

2a

2.3

4 ±

0.3

7a

1.2

0 ±

0.0

5a

53.4

3 ±

3.1

3a

269.1

7 ±

21.8

2a

P

0.0

752

0.0

370

0

.2699

0.1

438

0.0

683

0.2

587

0

.0862

Tab

le 3

. C

ente

sim

al

com

po

siti

on

an

d c

alo

ric

va

lue

mea

n a

nd

sta

nd

ard

dev

iati

on

on

acc

ou

nt

of

the

dry

mate

ria

l (e

xce

pt

for

the

mois

ture

con

ten

t) o

f th

e fo

ur

an

aly

zed

pu

rple

ya

m i

nco

rpo

rate

d b

read

fo

rmu

lati

on

s (0

%,

10

%,

15%

an

d 2

0%

). P

rob

ab

ilit

y (

P)

valu

es c

alc

ula

ted

fro

m K

rusk

al-

Wa

llis

AN

OV

A f

oll

ow

ed

by

post

hoc

test

s are

sh

ow

n.

Valu

es e

xh

ibit

ing

dif

fere

nt

lett

ers

in

th

e sa

me

colu

mn

in

dic

ate

sta

tist

icall

y s

ign

ific

an

t d

iffe

ren

ces

(P

<0.0

5).

statistical analysis through the statistical software

package (Statsoft STATISTICA 8.0 2007). Given to the

number of sampled observations (n=3), Kruskal-Wallis

ANOVA and post hoc tests were applied as a

non-parametric alternative to Fisher ANOVA, for

independent data, in the comparison among the bread

formulations.

Findings showing significance level of (P<0.05)

were considered as statistically significant.

Preparation of purple yam and its incorporated

breads methanolic extract

Samples of purple yam (Dioscorea trifida),

were peeled and ground with the aid of a knife. They

were then dehydrated in a laboratory oven at 60°C for

24 h. Purple yam incorporated breads of four

formulations: 0%, 10%, 15%, and 20%, were cut into

1 cm thick slices, and dehydrated in a laboratory oven at

40°C for 24 h (Hsu et al., 2004). Dehydrated yams and

breads were ground with pestle and mortar, weighed at

0.125, 0.25, 0.5 and 1.00 g (40, 80, 160 and 330 mg/mL,

respectively). They were placed into small test tubes

added with 5 mL of methanol and left in a rotary shaker

for 24 h. The material was centrifuged at 2,500 RPM

for 10 min so as to obtain the supernatant (methanolic

extract). The antioxidant activity of the samples was

determined by the free radical scavenging, 2.2-diphenyl-

1-picryl-hydrazyl (DPPH) and lipid peroxidation (LPO)

methods. The latter method evaluates the inhibition of

free radicals generated during the linoleic acid

peroxidation, and is based on spectrophotometric

measurements of discoloration (oxidation) of ß-carotene,

induced by linoleic acid oxidative degradation products

(Marco, 1968; Miller, 1971; Duarte-Almeida et al.,

2006).

Antioxidant activity determination through free

radicals scavenging methods (DPPH) in purple yam

and its incorporated breads

DPPH method, following the methodologies

described by Shimada et al., (1992) and Hsu et al.,

(2004), with some modifications, where 2 mg of DPPH

were dissolved into 15 mL of methanol, and applied so

as to determine the antioxidant activity of samples of

purple yam and its incorporated breads in the four

aforementioned formulations. A micro plate bearing

96 well was used. Thirty microliters (30 µL) of the

methanolic extract, plus 170 µL of methanol (used as the

blank) were placed in the wells. The reading was

performed on an Elisa reader (DXL 800-BECKMAN

COULTER) at a wavelength of 492 nm, using triplicate

samples. Then, 100 µL of the DPPH solution were

added, and the material was stored in a dark place for

30 min, and the reading was repeated as soon as this time

was over. Two hundred microliters (200 µL) of methanol

added to 100 µl of the DPPH solution were used as

the control. Thirty microliters (30 µL) of quercetin

(10 µg/mL), 170 µL of methanol and 100 µl of the

DPPH solution, were used as the standard. The following

formula was used so as to calculate the antioxidant

activity percentage

Antioxidant activity determination through the lipid

peroxidation (LPO) method in purple yam and its

incorporated breads

The determination of the antioxidant activity of

the samples through the LPO method was carried out

according to the method reported by Duarte-Almeida

et al., (2006), based on the methodology originally

described by Marco (1968), and later modified by Miller

(1971). The reactive mixture was prepared in

an Erlenmeyer flask, containing 50 µL of linoleic acid,

200 µL of tween 80 (emulsifying agent), 150 µL of

ß-carotene solution at 2 mg/mL in chloroform, and

500 µL of chloroform. The mixture was then subjected to

evaporation in nitrogen till there was no more

chloroform left. Later, the mixture of 25 mL of

previously oxygen saturated water was added, and during



A sample - A blank

% AA = 100 - x 100

A control

a period of 30 min it was homogenized through vigorous

shaking.

The reactive mixture showed to be clear with

absorbency ranging from 0.6 to 0.7 at a wavelength of

492 nm. A 96 well bearing micro plate was used. Two

hundred forty microliters (240 µL) of the reactive

mixture and 10 µL of the methanolic extract samples

were placed in the wells. Ten microliters (10 µL) of

methanol and an equal volume of butylhydroxytoluene

(BHT) at a concentration of 40 µg/mL were used as

control and standard, respectively. The micro plate was

incubated at 50ºC to speed up the oxidation reactions and

start β-carotene discoloration. Discoloration slope

readings of samples, control and BHT (in triplicate) were

performed readily, in an Elisa reader at a wavelength of

492 nm every 15 min for 135 min. The following

formula was used so as to calculate the oxidation

inhibition percentage:

Sensory analysis of purple yam incorporated breads

The acceptance test of purple yam in natura

incorporated breads counted with the participation of 78

non-trained volunteer judges. Each one of them was

provided with an answering card bearing a 9 point

hedonic scale (9-like extremely to 1-dislike extremely),

adapted from Stone et al., (1993) and Silva et al., (2005).

The judges were provided with three purple yam

incorporated bread samples, produced from three

formulations (10%, 15% and 20%) (Table 1). Samples

were served in white, disposable plastic plates; encoded

with three randomly chosen numbers. Samples were

evaluated according to their sensory qualities: global

feel, aroma, flavor, color and texture. Judges were

advised to always rinse their mouth with water before

testing the next sample.

The findings obtained on the acceptance test

were submitted to statistical analysis through statistical

software package (Statsoft STATISTICA 8.0 2007). The

Shapiro-Wilk test rejected the frequency distribution

normality of the three tested bread formulations, in all

their sensory attributes. However, the Levene test

accepted the homocedasticity (homogeneity of variances)

among the formulations for all sensory attributes. As

frequency distribution normality and variance

homogeneity are basic assumptions made for the

application of parametric tests, such as Fisher’s

ANOVA, and as these assumptions were not attended to,

the Friedman ANOVA followed by post hoc tests were

applied as a non-parametric alternative for paired data in

bread comparisons. Findings presenting significance

level of (P<0.05) were considered as statistically

significant.

Microbiological analysis of purple yam breads

Following the recommendation of the

Brazilian National Health Surveillance Agency

(in Portuguese, Agência Nacional de Vigilância

Sanitária, ANVISA), based on Ruling Number 12 (RDC,

2001), we carried out the microbiological analysis so as

to verify Coliforms and Salmonella in samples of the

three purple yam bread formulation samples through the

membrane filtration method (APHA, 2001).

RESULTS AND DISCUSSION

Centesimal composition and caloric value of purple

yam

Moisture (76.43±0.50), protein (1.83±0.13) and

ash (0.78±0.02) contents, as well as the caloric value

(89.64±4.52) of purple yam (D. trifida) samples analyzed

in the present study (Table 2) show to be near

those presented by Montaldo (1991) for yam

(Dioscorea spp.) and those found in the Brazilian

Food Composition Table TACO (2006), for the yam

(D. alata). Lipid content (1.13±0.69) stayed well above

that presented by Montaldo (1991) and TACO (2006).

Crude fiber content (1.80±0.05) is above the value

observed by Montaldo (1991), and well below that



A2 sample - A1 sample

% I = 100 - x 100

A2 control - A1 control

presented in TACO (2006). The high fiber content

presented by TACO (2006) might be due to the

enzymatic gravimetric method employed in the analyses.

That method warrants a higher precision for determining

the dietary fiber as compared to the acid digestion

methodology used in the present study as well as

by Montaldo (1991). Total carbohydrate content

(18.04±0.66) is well below Montaldo (1991) and TACO

(2006) values. The remaining differences in centesimal

composition values presented by Montaldo (1991) and in

the present study might be related to the different soil

types being employed on planting the tubers and/or to the

different species being utilized. Nevertheless, the

different values presented in TACO (2006) may be

related to the different yam species being analyzed.

Centesimal composition and caloric value of purple

yam incorporated breads

Based on data from Kruskal-Wallis (ANOVA)

followed by post hoc tests (Table 3), it may be asserted

that, except for the lipids (P<0.05), all other centesimal

composition and caloric values of the four purple yam

incorporated bread formulations (0%, 10%, 15% and

20%) showed to be statistically similar (P>0.05). That is,

replacing wheat flour by purple yam in natura in up to

20% neither modifies bread centesimal composition nor

caloric value. As for lipid, statistically significant

difference was only observed for 10% and 20%

formulations; this negligible 0.5% difference may be

neglected in technological applications.

Purple yam incorporated breads centesimal

composition and caloric value were compared to those of

ordinary bread loaf (OBL) (Anton et al., 2006) and

whole bread loaf (WBL) (TACO, 2006) (Table 4). One

notices, a high fiber content (6.90%) in the whole bread

loaf (WBL) (TACO, 2006), relative to the remaining

breads. It can be highlighted that in whole bread

composition, we have the presence of grain-composed

whole flour, almost wholly made up of bran, germ and

endosperm (FDA, 2006). By and large, all other values

show to be approximate. All differences found may be

related to formulations employed in the preparation of

those breads.

Antioxidant activity determination through the free

radical scavenging method (DPPH) in purple yam

and its incorporated breads

Antioxidant activity (% AA) of the methanolic

extract pertaining to purple yam (Dioscorea trifida)



Bread Moisture

(%)

Lipid

(%)

Protein

(%)

Crude Fiber

(%)

Ash

(%)

Carbohydrate

(%)

Caloric value

(kcal/100 g)

0% 29.79 4.49 11.62 1.95 1.18 50.95 290.73

10% 31.03 4.24 10.67 1.91 1.41 53.41 294.45

15% 32.65 4.87 9.82 1.84 1.38 49.45 280.64

20% 35.09 4.74 10.06 2.34 1.20 53.43 269.17

OBL 34.46 1.93 9.42 2.57 2.09 52.10 247.50

WBL 34.70 3.70 9.40 6.90 2.30 49.90 253.00

Table 4. Centesimal composition and caloric value of ordinary bread loaf (OBL) (Anton et al., 2006),

whole bread loaf (WBL) (TACO, 2006) and purple yam (D. trifida) incorporated breads at 0%, 10%,

15% and 20% (present study).

0

10

20

30

40

50

60

70

80

90

100

40 80 160 330

Concentration (mg/mL)

An

tio

xid

an

t acti

vit

y (

%)

Purple yam

0% Bread

10% Bread

15% Bread

20% Bread

Quercetin

Figure 2. Antioxidant activity expressed by free

radical scavenging percentage, of samples of purple

yam (Dioscorea trifida) and its incorporated bread

extracts in four formulations: 0%, 10%, 15% and

20%, as determined by DPPH method. The quercetin

was used as standard control. Bars indicate standard

deviation.

samples in the concentrations of 330, 160, 80 and

40 mg/mL, as determined by the DPPH method, were

higher than 70%, reaching a maximum of 88.13±0.12.

This plainly shows this species to exert DPPH radical

scavenging activity (Figure 2). This same figure reveals

purple yam incorporated breads prepared in 10%, 15%

and 20% formulations, to also present a certain

antioxidant activity, reaching 43.32±1.18; 48.13±1.17

and 53.71±1.01 maximum percentile values,

respectively. Those findings are above the values

presented by Hsu et al., (2004) (20-40% approximately),

who used breads of several formulations prepared with

flour from the purple yam tuber (Dioscorea purpurea)

representing the one with the widest variety in Taiwan,

for substituting part of the wheat flour. Bread prepared

with no purple yam at all (0%) showed certain

antioxidant activity, as well, probably due to Maillard

reaction products, where, some hot processed foods,

present free radical scavenging activity (Kim et al.,

2007; Jing and Kitts, 2000; Hsu et al., 2004; Michalska

et al., 2008). Corroborating data from Hsu et al., (2004),

it was confirmed that the antioxidant activity rose as the

percentage of purple yam substituting wheat flour

increased. The high free radical scavenging activity

observed by Hsu et al., (2004) in flour of Taiwan purple

yam (D. purpurea), was also detected in the Amazonian

region’s purple yam (D. trifida).

Antioxidant activity determination through the lipid

per oxidation (LPO) method in purple yam and its

incorporated breads

Discoloration slope (Figure 3) and free radical

inhibition activity (Figure 4) determined through the

LPO method confirmed the antioxidant activity (% I) in

purple yam (55.80±4.85) and its breads from the three

formulations (10%, 15% and 20%), with the values of

46.16±4.90; 48.20±3.72 and 49.13±2.79, respectively.



Tests Bread% Color Aroma Flavor Texture Overall impression

Shapiro-Wilk P 10 0.0009 < 0.0001 0.0023 0.0044 0.0001

15 < 0.0001 0.0009 0.0008 0.0019 < 0.0001

20 < 0.0001 0.0001 0.0002 0.0090 0.0005

Levene P 0.0519 0.5580 0.2306 0.8415 0.5184

Table 5. Probability (P) values calculated from Shapiro-Wilk and Levene tests for evaluating frequency

normality and homogeneity of variances, respectively, of the data obtained in the sensory analysis of the

three tested purple yam incorporated bread formulations.

Values were considered statistically significant at (P< 0.05).

0

0.1

0.2

0.3

0.4

0.5

0.6

0.7

0.8

0.9

0 15 30 45 60 75 90 105 120 135

Time (min)

Ab

s 4

92

Blank

Purple yam

0% Bread

10% Bread

15% Bread

20% Bread

BHT

Figure 3. Discoloration slope of purple yam

(Dioscorea trifida) and its incorporated bread

extracts in four formulations: 0%, 10%, 15%, 20%,

blank and BHT, as determined through the LPO

method.

0

10

20

30

40

50

60

70

80

90

Purple yam 0% Bread 10% Bread 15% Bread 20% Bread BHT

Inh

ibit

ion

(%

)

Figure 4. Inhibition percentage of free radicals of

purple yam (Dioscorea trifida) and its incorporated

bread extracts in four formulations: 0%, 10%, 15%,

20%, and BHT as determined by LPO method. Bars

indicate the standard deviations.

As it was observed by the DPPH method, the

antioxidant activity rose as the percentage of purple yam

substituting wheat flour in the breads increased.

Moreover, bread with no addition of purple yam (0%)

presented some antioxidant ability which might have

resulted from the development of Maillard reaction

products (Kim et al., 2007; Jing and Kitts, 2000; Hsu

et al., 2004; Michalska et al., 2008). Anthocyanins might

be partly responsible for the antioxidant activities

detected in the purple yam (D. trifida) and its

incorporated breads analyzed in the present study, since

these pigments were detected in purple yams, D. alata

(Rasper and Coursey, 1967) and D. trifida L.

(Carreno-Diaz and Grau, 1977; Escudero et al., 2010).

In fact, polyphenols and anthocyanins, usually detected

in plants, might be the active components for this

antioxidant activity in yams (Hou et al., 2001; Hsu et al.,

2004).

Sensory analysis of purple yam incorporated breads

Table 5 shows the rejection of the frequency

distribution normality of the three tested purple yam

incorporated bread formulations (10%, 15% e 20%)

through the Shapiro-Wilk test, and the acceptance of the

homocedasticity among the formulations through the

Levene test on all sensory attributes evaluated in the

acceptance test (i.e. statistically significant values at

(P <0.05).

Friedman ANOVA followed by post hoc tests

applied for comparing the three purple yam incorporated

bread formulations revealed a significant difference

(P<0.05), only for the colour attribute (Table 6). The

bread at 20% presented a better evaluation regarding the

remaining ones, probably due to the higher purple yam

concentration, which gives the final product a more

attractive kind of color. It was observed that the larger

the purple yam amount being added to the bread the

higher the mean score obtained (values ranging from

6.15 to 6.97).

Choosing a determined food should depend



Bre

ad

Colo

r A

rom

a

Fla

vo

r

Textu

re

Over

all

im

pre

ssio

n

Mea

n

Med

ian

M

ean

M

edia

n

Mea

n

Med

ian

M

ean

M

edia

n

Mea

n

Med

ian

10%

6.1

5 ±

1.4

6ª

6

6.7

1 ±

1.2

2ª

6

6.2

8 ±

1.4

4ª

6

6.0

3 ±

1.5

0ª

6

6.5

4 ±

1.2

3ª

6

15%

6.2

9 ±

1.3

0ab

6

6.5

1 ±

1.3

4ª

6

6.4

4 ±

1.2

2ª

6

5.9

4 ±

1.6

7ª

6

6.5

9 ±

1.1

4ª

6

20%

6.9

7 ±

1.5

5b

7

6.6

8 ±

1.2

0ª

6,5

6.4

1 ±

1.4

3ª

6

6.2

2 ±

1.6

1ª

6

6.8

8 ±

1.3

3ª

7

P

0.0

001

0.6

285

0.9

641

0.5

264

0.0

608

Tab

le 6

. S

enso

ry e

va

luati

on

res

ult

s of

the

thre

e p

urp

le y

am

in

corp

ora

ted

bre

ad

fo

rmu

lati

on

s. P

rob

ab

ilit

y (

P)

valu

e w

as

ob

tain

ed t

hro

ugh

Fri

edm

an

AN

OV

A f

oll

ow

ed b

y p

ost

hoc

test

.

Valu

es e

xh

ibit

ing d

iffe

ren

t le

tters

in

th

e sa

me

colu

mn

poin

t ou

t st

ati

stic

all

y s

ign

ific

an

t d

iffe

ren

ces

(P<

0.0

5).

mainly on its nutritional value. Nevertheless, color,

aroma and texture are the factors usually guiding the

consumer’s preference rate. Of these three factors, color

interferes the most on the product’s preference (Bobbio

and Bobbio, 2001).

Given that there were no preferential differences

among the other sensory attributes, the three breads

evaluated can be considered approved.

Microbiological analysis

Considering that the microbiological analysis

was negative for Coliforms and Salmonella (Table 7), the

purple yam incorporated breads may be considered

proper for human consumption, as long as they have

been properly handled.

CONCLUSIONS

Through such findings, one concludes that any

of the purple yam incorporated breads tested in the

present study (10%, 15% and 20%), can substitute

ordinary bread (0%), with no effect on the diet’s caloric

value, since their centesimal compositions are similar.

Due to the presence of antioxidants in purple yam

incorporated bread, and to their ability to fight free

radicals, those breads can be considered as

health-promoting food. Furthermore, these findings point

out the feasibility of the consumption of purple yam

incorporated bread, as an alternative in the local bread

making industry, and an incentive to a larger production

of this tuber in the Amazonian region.

ACKNOWLEDGEMENTS

The authors are indebted to the Fundação de

Amparo à Pesquisa do Estado do Amazonas (FAPEAM)

for the master scholarship granted to Antonia Paiva

Teixeira. To Dr. Antonio José Inhamuns da Silva and

MSc. Cynthia Tereza Corrêa da Silva of Universidade

Federal do Amazonas (UFAM) for kindly having

allowed to carry out the centesimal composition analyses

in their laboratories. To MSc. Antonio Fábio Lopes de

Sousa, Arleilson de Sousa Lima and Ana Cláudia dos

Santos for their invaluable aid in undertaking of the

laboratory analyses. To Misters Claudio Adriano

Cardoso Amanajás and Francisco de Oliveira Batista for

their logistical support in the collection of purple yam

samples.

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http://www.sciencedirect.com/science/journal/07335210

Jou

rn

al of R

esearch

in

Biology

Bioefficacy of Novaluron®, a chitin synthesis inhibitor against the tropical

warehouse moth, Ephestia cautella.

Keywords: Novaluron, Hocklicombi®, Ephestia cautella, warehouse moth, chitin, loss assessment.

ABSTRACT: The tropical warehouse moth, Ephestia cautella (Lepidoptera: Pyralidae) is a major pest of stored maize in Ghana. It is controlled mainly by the use of synthetic insecticides which has become a major challenge in the stored product industry in Ghana. Both laboratory and field trials were conducted to evaluate the efficacy of novaluron, a chitin synthesis inhibitor against E. cautella. Five concentrations of Novaluron (0.1, 0.2, 0.3, 0.4 and 0.5 mL/L of water) were prepared and each concentration was topically applied on the notal regions of 10 fifth instar larvae of E. cautella per concentration. At 0.4 mL/L and 0.5 mL/L treatments, larval mortality ranged between 50-80% after 96 h of exposure. Also, Novaluron (0.5 mL/L) was used to treat four surfaces (concrete, wood, glass and plastic) usually encountered in structural insect pest management systems and the larvae exposed to these surfaces. Hocklicombi® (5 mL/L) served as positive control. Larval mortality (35.5-97.5%), pupation (0.0-35.0%) and adult emergence (0.0-20.0%) in surfaces treated with Hocklicombi® compared favourably with those treated with Novaluron (25.0-97.5%), (2.5-60%) and (0.0-42.5%), respectively. A simulated field experiment was conducted in which four batches of 5 kg of maize in miniature bags were pretreated with 0.4 mL/L Novaluron and 50 unsexed adults were introduced. This was left in a crib at the University of Ghana farm for 60 days. The field experiment showed that after 60 days of storage there was a lower weight loss in the Hocklicombi® (6.6%) and Novaluron (6.8%) treatments compared to the negative control (11.3%).

759-767 | JRB | 2013 | Vol 3 | No 1

This Open Access article is governed by the Creative Commons Attribution License (http://

creativecommons.org/licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.

www.jresearchbiology.com Journal of Research in Biology


Research Journal

Authors:

Sackey I, Eziah VY and

Obeng-Ofori D.

Institution:

Department of Crop Science,

College of Agriculture and

Consumer Sciences, P. O.

Box LG 44, University of

Ghana, Legon.


Eziah VY.

Web Address:

http://jresearchbiology.com/

documents/RA0305.pdf.


Article Citation: Sackey I, Eziah VY and Obeng-Ofori D. Bioefficacy of Novaluron®, a chitin synthesis inhibitor against the tropical warehouse moth, Ephestia cautella. Journal of Research in Biology (2013) 3(1): 759-767


Original Research

INTRODUCTION

Maize (Zea mays) is one of the major staple food

crops in Ghana and it is susceptible to attack by several

insect pests including the tropical warehouse moth,

Ephestia cautella Walker (Lepidoptera: Pyralidae)

(CAB1, 2006). Ephestia cautella larva feeds on stored

products, damaging the product directly and form webs

on the surface. The webbing contains larval excreta and

exuviae which give unpleasant odour to the infested

commodity. Older larvae may leave the food to find

pupation sites in wall cracks.

In Ghana, E. cautella is controlled by the use of

residual insecticides usually, synthetic pyrethroids and

fumigants (CABI, 2006). The adverse effects of residual

pesticides such as poisoning, environmental and health

hazards and resistance development cannot be

overemphasized (Obeng-Ofori, 2007). Hence the use of

residual insecticides in stored product protection is

challenging. There is therefore, the need for new cost and

environment friendly alternatives with no adverse effect

on non-target organisms (Obeng-Ofori, 2007; Arthur and

Phillips, 2003). Some of these alternatives include

botanicals, insect growth regulators, microbial pathogens

among others (Arthur, 1996).

Novaluron (Rimon® 10 EC) is a benzoylphenyl

urea group of insect growth regulators and a chitin

synthesis inhibitor. Novaluron has been registered as an

insecticide for food crops in several countries including

South Africa, Australia and Ghana (WHO, 2003; EPA,

2006). In Ghana, novaluron has been successfully used

in the laboratory against stored product pests

such as the rice moth, Corcyra cephalonica

Stainton (Lepidoptera: Pyralidae) (Sarbah, 2006), the

red flour beetle, Tribolium castaneum Herbst

(Coleoptera: Tenebrionidae) (Bakudie, 2006) and the

tropical warehouse moth, E. cautella (Ibrahim, 2008).

In Ghana, most of the work done on novaluron

focused on evaluating the effect of the chemical on

different developmental stages of insects in the

laboratory. There is no information on the use of

novaluron for stored product protection in warehouses or

cribs. Ephestia cautella is noted for feeding directly on

the grains and also, the mature larvae leave the

commodity in search of pupation sites in crevices, cracks

and storage containers. Therefore, treating these surfaces

to which the insect may be exposed will go a long way to

mitigate the losses caused by this pest. Hence, screening

Novaluron against E. cautella using the commodity and

storage surfaces as substrates is crucial in the

management of the pest. Such findings will contribute to

the efforts by farmers and warehouse managers to reduce

storage losses and contribute to the attainment of food

security in Ghana.

This study presents laboratory and field tests that

were carried out to determine the toxicity of novaluron to

the 5th instar larvae of E. cautella. Other tests were also

conducted to assess the efficacy of novaluron on

different surfaces against immature stages of E. cautella.


Insect cultures

The test insects were obtained from the

Entomology laboratory of the Crop Science Department.

Adult E. cautella were cultured on mixed substrate made

up of wheat powder, maize flour and glycerol (5:5:1).

Fifty adult E. cautella were introduced into each jar and

left under laboratory conditions of 27±2°C and 55-60%

relative humidity for 30 days to allow for the

development of larval E. cautella. The set up was placed

on trays containing industrial oil to prevent the crawling

of other insects into the culture. The insects were reared

and handled using ethically acceptable standard

procedures in the laboratory.

Test chemicals

Novaluron (Rimon® 10EC), 1-[3-chloro-4-(1, 1,

2-trifluoro-2-trifluoromethoxy-ethoxy) phenyl]-3-(2, 6

difluorobenzoyl) urea, produced by Makhteshim-Agan

Ltd (Israel) was used for the toxicity experiment and

Sackey et al., 2013


Hocklicombi (Hockley International Ltd. Poynton,

Stockport, U. K.) which contains 25% Fenitrothion and

5% Fenvalerate was used as a reference product.

Contact toxicity test

We adopted the method by (Eziah et al., 2011).

Concentrations of Novaluron (0.1, 0.2, 0.3, 0.4 and

0.5 mL/L) and 5 mL/L of Hocklicombi® were diluted in

distilled water and used for the assays. Distilled water

was used as negative control. Fifth instar larvae of both

sexes were transferred into clean Petri dishes and the

different dosages of the various concentrations was

topically (1 µL) applied to the notal regions of the larvae

using a micro applicator. Each experimental unit

consisted of 10 larvae and was replicated for four times.

The treated insect larvae were then transferred into glass

petri dish containing food. The insect larvae were

examined for mortality 24, 48, 72, 96 h, 7 days and

14 days after treatment. Criterion for death was as

described by (Lloyd, 1969) in which insects were

presumed dead when they failed to move in a

coordinated manner after prodding with a blunt probe.

Data collected include larval mortality, percent pupation

and percent adult emergence were done after various

treatments and exposure periods.

Surface treatment

The surfaces chosen for the study were concrete,

plywood, glass and plastic which are among the

common surfaces encountered in structural insect pest

management. Individual concrete exposure arenas were

created in square bottoms of plastic containers (6x6 cm)

using a concrete patching material. Water-based slurry

was prepared by mixing 1 kg of Portland cement to 2 kg

of sand and 1 L of tap water and pouring 10 mL of the

slurry into the bottom of the plastic container to create a

treatment arena (Arthur, 1998b). Plywood arenas were

made by cutting rectangular disks from 1.25 cm thick

plywood to fit the plastic container then caulking the

margins to prevent the larvae from escaping the surface.

Plastic containers served as plastic surfaces and petri

dishes were used as glass surfaces for the surface

treatment.

Each of the four surfaces was treated with 4 mL

of water (negative control treatment) or an aqueous

solution of novaluron (0.5 mL/L) and Hocklicombi®

(5 mL/L) (positive control). All treated arenas were

allowed to dry overnight and fifth instar larvae (N=10) of

E. cautella were exposed for 48 h. The larvae were then

transferred to new petri dishes containing food under

laboratory conditions of 27±2°C and 55-60% relative

humidity. Post-treatment survival and mortality were

recorded daily. Number of surviving larvae that

successfully pupated and those that successfully emerged

as adults were recorded.

Field experiment

Maize grains were obtained from the Madina

(a suburb of Accra, Ghana) market and sieved to remove

all debris. Maize grains (5 kg) were sterilized in the oven

at 70°C for 3 h after which they were left in desiccators

to cool. The grains were then treated with 0.4 mL/L

Novaluron or 5 mL/L Hocklicombi®. These dosages had

proven effective in laboratory experiments. Grains

treated with distilled water served as negative control.

Each treatment was replicated four times. Fifty unsexed

adults of E. cautella were put onto the treated grains in

each sack. The sacks were securely sealed by stitching

and stored in a grain crib at the University farm for

60 days. Prior to their treatment, subsamples were taken

from each sack for moisture and weight loss analyses

using the standard volume method was carried out

(Boxall, 1986). At the end of the storage period, the

contents of the sacks were sieved. The number of

both live and dead adult insects was recorded. Also,

subsamples of the maize grains were collected for

moisture and weight loss analyses as stated earlier.

Statistical analysis

Data involving percentages were arcsine

transformed and were analyzed using the Analysis

of Variance (ANOVA) with Genstat 9.2

Sackey et al., 2013


(Lawes Agricultural Trust, 2007). Means were separated

using the Least Significant Difference (LSD) test at

5% probability level.

RESULTS

Contact toxicity test

The percent larval E. cautella mortality

following treatment with Novaluron and Hocklicombi®

are presented in Table-1. Larval mortality varied with

insecticide concentration and exposure period. Lower

dosages of Novaluron (0.1-0.3 mL/L) caused less

than 50% larval mortality after 96 h of exposure

(Table 1). In contrast, novaluron concentrations of

0.4 mL/L and 0.5 mL/L caused between 50% to 80%

larval mortality after 72 to 96 h of exposure. After 96 h

exposure period, all dosages of Novaluron induced

significantly (p = 0.05) higher larval mortality compared

to the negative control. However, there was no

significant difference in larval mortality between 5 mL/L

Hocklicombi® and 0.5 mL/L Novaluron treatments. Also,

novaluron applied at 0.4 ml/L and 0.5 mL/L did not

differ significantly from each other after 96 h of

exposure.

Pupation and adult emergence of E. cautella

were observed in all insecticide treatments and the

negative control with the exception of Hocklicombi®

treatment. The percentage pupation in larvae treated with

0.1 mL/L Novaluron (57.5-65.0%) was not significantly

different from the negative control (64.0-80.0%)

(Figure 1). However, all other concentrations of

Novaluron higher than 0.1 mL/L significantly (p = 0.05)

impaired pupation. There was no significant difference in

pupation 7 days after the exposure of E. cautella larvae

to 0.5 mL/L Novaluron and 5 mL/L Hocklicombi®.

Also, after 14 days, percentage pupation recorded in

larvae treated with 0.4 mL/L and 0.5 ml/L was

comparable.

All levels of Novaluron concentrations

significantly reduced the development of F1 of adult

E. cautella (Figure 2). The highest adult emergence

(77.5%) was recorded in the negative control and this

differed significantly (p = 0.05) from all other novaluron

concentrations applied. As concentration increased from

0.1 mL/L to 0.5 mL/L, adult emergence significantly

(p = 0.05) reduced from 50 to 2.5%. Also, the effect of

novaluron applied at 0.5 mL/L was comparable to

Hocklicombi® treatment in impairing the development of

adult E. cautella.

Surface treatment

Ephestia cautella larvae exposed on concrete

surfaces treated with Novaluron showed a lower

mortality than those exposed to concrete surfaces treated

with Hocklicombi® (Table 2). Mortality was also lower

on plastic and wood treated surfaces compared to

Hocklicombi® treated surfaces. However, the percentage

mortality of E. cautella on glass surfaces treated with

Sackey et al., 2013


Treatments (ml/L) Mean±(s.e) % larval mortality (h)

24 h 48 h 72 h 96 h

Control (Water) 0.0±0.0 0.0±0.0 0.0±0.0 0.0±0.0

5.0 mL/L (HC) 87.5±0.1 87.5±0.1 87.5±0.1 87.5±0.1

Novaluron

0.1 7.5±0.0 10.0±0.0 17.5±0.1 22.5±0.1

0.2 10.0±0.0 12.5±0.0 27.5±0.1 42.5±0.1

0.3 17.5±0.1 25.0±0.1 35.0±0.1 45.0±0.2

0.4 17.5±0.1 32.5±0.1 50.0±0.1 66.0±0.1

0.5 32.5±0.1 47.5±0.1 65.0±0.1 80.0±0.0

LSD (P < 0.05) 18.35 18.40 14.50 14.83

HC= Hocklicombi®

s.e = standard error

Table 1 Mortality of larval E. cautella (%) after treatment

with novaluron and Hocklicombi® insecticides

novaluron was the same (97.5%) as those treated with

Hocklicombi®. Surviving larvae were observed for

pupation and adult emergence. Fewer E. cautella larvae

pupated after exposure to concrete, plastic and wood

surfaces treated with Hocklicombi® but no pupation was

recorded on glass surfaces treated with Hocklicombi®

(Table 3)

Fewer larvae pupated in glass surfaces-treated

with novaluron and this was not significantly different

from Hocklicombi®-treated glass surfaces. Generally,

percentage adult E. cautella that emerged was greater on

the untreated control for all the surfaces and differed

significantly (p = 0.05) from all insecticide treated

surfaces (Table 4). Mean percentage adult emergence of

E. cautela observed on glass and plastic surfaces treated

with novaluron and Hocklicombi® ranged from 0.0 to

25%. Thus, residual effects of novaluron and

Hocklicombi® significantly reduced the development of

E. cautella on glass and plastic surfaces.

Field experiment

Table 5 shows the dry weight loss of the treated

grains after 60 days of storage using the standard volume

method. Lower weight losses were observed in grains

treated with insecticides (6.6-6.8%) compared to grains

which were not treated (11.3%) using standard volume

methods.

DISCUSSION

The present study showed that Novaluron

concentrations of 0.4 mL/L and 0.5 mL/L significantly

affected the metamorphosis of E. cautella to the adult

stage. The effectiveness of novaluron at these dosages

compared favourably with Hocklicombi®. The insect

growth regulator’s ability to regulate metamorphosis in

the larvae through contact by topical application is

consistent with its mode of action. Tomlin (2005)

reported that novaluron was very effective on the larvae

of insects when absorbed by ingestion and contact

activity. The author also reported that the compound

causes abnormal endocuticular deposition and abortive

moulting.

Although pupation and adult emergence were

observed in all treatment levels, most of the larvae

treated with 0.4 mL/L and 0.5 mL/L Novaluron could not

emerge into adults 23 days after treatment. This may be

attributed to abnormal endocuticular deposition and

abortive moulting in the larvae (Tomlin, 2005). Also,

when cocoon covering the pupae were slightly removed,

pupae found were malformed compared to those in the


Sackey et al., 2013

Mean (%) ± s.e mortality

Insecticide Type of surface

Concrete Glass Plastic Wood Means

Control 0.0 ± 0.0 0.0±0.0 2.5 ± 0.0 0.0 ± 0.0 0.7±0.0

Hocklicombi® 43.0 ± 0.1 97.5±0.0 45.0 ± 0.1 35.0 ± 0.1 55.0±0.0

Novaluron 25.0±0.1 97.5±0.0 17.5 ± 0.1 25.0 ± 0.1 41.1±0.0 Means 22.7± 0.0 64.7±0.0 21.7 ± 0.0 20.0 ± 0.0 -

Table 2 Mortality of E. cautella larvae (%) after 7 days exposure on concrete, glass,

plastic and wood surfaces treated with Hocklicombi® and novaluron insecticides

LSD(P < 0.05): Main effects (insecticide = 1.21, surface= 1.39 Interaction (insecticide x surface)=2.4

Means (%) ± s.e for pupation



Control 92.5±0.0 95.0±0.0 95.0±0.0 97.5±0.0 95.0±0.0

Hocklicombi 35.0±0.1 0.0±0.0 35.0±0.1 25.0±0.0 23.8±0.0

Novaluron 55.0±0.1 2.5±0.0 60.0±0.1 47.5±0.1 43.1±0.0

Means 61.9±0.0 41.2±0.0 63.1±0.0 56.2±0.0 -

Table 3: Percentage pupation of E. cautella after 14 days exposure on concrete, glass, plastic and wood surfaces treated with Hocklicombi® and novaluron insecticides

LSD(P < 0.05): Main effects (insecticide5.6, surface= 6.6) Interaction (insecticide x surface)=13.20

control. Adults that emerged were found not to be active

as those in the control. These findings are consistent with

reports by Amos and Williams (1974). According to

CABI (2006), pupal formation is completed in seven

days and development from egg to adult ranges from

29-31 days under optimum conditions of 32.5oC and

70% relative humidity. However, in the present study

under laboratory conditions of 27±2°C and 55-60%

relative humidity, pupation extended up to 14 days and

adult emergence was also delayed up to 30 days in the

treated 5th instar larvae of E. cautella. Thus, novaluron

was found to prolong the development period of

E. cautella larvae to adults.

The ability of Novaluron to reduce the number of

new generations is consistent with the findings of

(Kostyukovsky et al., 2003) and Kostyukovsky and

Trostanetsky (2006). The authors found that novaluron

applied at 1 ppm reduced the number of new generations

of S. oryzae and R. dominica by 95% and also caused

total mortality of the 3rd instar larvae of T. castaneum.

The effectiveness of novaluron in preventing the

metamorphosis of E. cautella when applied at 0.4 mL/L

and 0.5 mL/L also confirms work done by Ibrahim

(2008). The author found that development of E. cautella

to adults was prevented when novaluron was applied at

0.4 mL/L and 0.6 mL/L.

These observations indicate that the effectiveness

of novaluron as a grain protectant depends on the species

of insect, dosage and exposure time. Wilson and Cryan

(1997) and Mulla et al., (2003) stated that the effects of

chitin synthesis inhibitors vary according to species,

development stage, time of application, kind of

compound and dose administered.

The residual effect of Hocklicombi® and

Novaluron were significantly greater on glass surfaces

than plastic, concrete or wood surfaces. Generally,

Hockicombi® significantly caused higher mortalities on

all the surfaces than novaluron. The high residual

efficacy of Hocklicombi® may be attributed to the

components of the compound. Hocklicombi® contains

fenitrothion and fenvalerate as its active ingredients.

These compounds have been reported by several

researchers to have high residual effects when used as

surface treatment against storage insects (Orui, 2004).

Both compounds are non-systemic insecticides with

contact and stomach activity (Tomlin, 2005).

In the present study, novaluron demonstrated

excellent residual effect on glass surfaces by preventing

the metamorphosis of E. cautella to the adult stage. The

residual effect on glass surfaces treated with novaluron

compared well with Hocklicombi®. However, on plastic,

concrete and wood surfaces, Novaluron was less

effective compared with Hocklicombi® but differed

significantly from the untreated control surfaces.

However, the residual effectiveness on plastic surfaces

showed better efficacy than on concrete and wood

surfaces.

The excellent effectiveness of Novaluron on

glass and plastic surfaces is consistent with work done by

(Atkinson et al., 1992). The authors found that when

hydropene, an insect growth regulator was sprayed on

non-absorbent surfaces such as glass and ceramic tile, the

Sackey et al., 2013


Means (%) ± s.e for adult emergence



Control 92.5±0.0 90.0±0.0 95.0±0.0 97.5±0.0 93.8±0.0

Hocklicombi 20.0±0.1 0.0±0.0 12.5±0.1 12.5±0.0 11.2±0.0

Novaluron 42.5±0.1 0.0±0.0 25.0±0.1 42.5±0.1 27.5±0.0 Means 50.6±0.0 32.5±0.0 45.0±0.0 48.1±0.0

LSD(P < 0.05): Main effects (insecticide5.9, surface= 6.34)= 6.34 Interaction Insecticide x surface=12.67

Table 4 Percentage adult emergence of E. cautella after 30 days exposure on concrete,

glass, plastic and wood surfaces treated with Hocklicombi® and novaluron insecticides

survival, number of oothecae and percentage of

cockroaches were more affected than on absorbent

surfaces of finished plywood and fibreboard. The low

mortality rates, pupation and adult E. cautella that

emerged after exposure to concrete and wood surfaces in

the current study can also be attributed to the

composition of these surfaces. Burkholder and Dicke

(1966) reported that new concrete surfaces contain high

levels of alkaline which hydrolyze residues and reduce

residual efficacy of insecticides hence, the low mortality

rates on concrete-treated surface in the present study was

not unexpected. Chadwick (1985) attributed low efficacy

of insecticides on plywood surfaces to vaporization,

chemical degradation, photodegradation and absorption

of insecticides into surfaces. Thus, the low mortalities

and higher survival rates observed in E. cautella exposed

to wood surfaces treated with the insecticides may be due

to the absorption of the insecticide into the wood

surfaces after treatment.

In the field experiment, all the insecticide

treatments significantly reduced dry weight loss in the

grains compared to the control. Novaluron was observed

to significantly reduce insect numbers in the treated

grains and also had a significantly lower dry weight loss.

Results from this study showed that novaluron

effectively protected maize grains from damage by

E. cautella. Grain weight losses calculated in the

Novaluron treatment compared well with those observed

in grains treated with Hocklicombi®. Considering that

Novaluron selectively targets larval stages by inhibiting

chitin synthesis and therefore, minimizes its impact on

adults of non targeted insect species (Ishaaya et al.,

2001), Novaluron can be used in replacement of residual

insecticides like Hocklicombi® for treatment of maize

grains for storage.

CONCLUSION

The current study showed that Novaluron was

effective in controlling the tropical warehouse moth. The

application of Nuvaluron at 0.4 mL/L and 0.5 mL/L

treatments resulted in larval mortality ranging between

50-80% after 96 h of exposure. Also, the treatment of

concrete, wood, glass and plastic surfaces usually

encountered in structural insect pest management

systems with 0.5 mL/L Novaluron induced (25.0-97.5%)

larval mortality, (2.5-60%) pupation and ((0.0-42.5%)

adult emergence. These figures were comparable to

those obtained from surfaces treated with 5 mL/L

Sackey et al., 2013


Figure 1 Percentage pupation (means±s.e) of

E. cautella larvae after treatment with novaluron and

Hocklicombi® insecticides. d = days h= hours

Figure 2 Percentage adult emergence (means±s.e) of

E. cautella after treatment with novaluron and

Hocklicombi® insecticides. d = days

Dosage (mL/L) Mean dry weight loss (%)

Control 11.3±0.0

Hocklicombi 5 6.6±0.0

Novaluron 0.4 6.8±0.0

Table 5 Percent dry weight loss after 60 days of

storage using the standard volume method

LSD(P < 0.05) = 1.63

Hocklicombi® insecticide. In the field maize treated with

0.4 mL/L Novaluron® and infested with adult E. cautella

after 60 days of storage showed that there was a lower

weight loss in the Hocklicombi® (6.6%) and novaluron

(6.8%) treatments compared to the negative control

(11.3%). This work has proven that Novaluron® could

replace the synthetic insecticides that are used in the

management of this pest and should be included in the

management programmes for storage pests control.

REFERENCES

Amos TG, Williams P, Du Guesclin B, Schwarz M.

1974. Compounds related to juvenile hormone: Activity

of selected terpenoids on Tribolium castaneum and

Tribolium confusum. J. Econ. Entomol., 67(4):474-476.

Amos TG. 1977. Williams P. Insect growth regulators:

Some effects of methoprene and hydropene on

productivity of several stored grain insects. Aus. J. Zool.,

25(2):201-206.

Arthur FH. 1996. Grain Protectants: Current status and

prospects for the future. J. Stored Prod. Res.,

32(4):293-302.

Arthur FH and Phillips TW. 2003. Stored product

insect pest management and control. In: Hui YH,

Bruinsma BL, Gorham JR, Nip WK, Tong PS, Ventresca

P. (eds.), Food Plant Sanitation, Marcel Dekker,

New York 341-358.

Arthur FH. 1998b. Effects of a food source on red flour

beetle (Coleoptera: Tenebrionidae) survival after

exposure on concrete treated with cyfluthrin. J. Econ.

Entomol., 91(6):773-778.

Atkinson TH, Koehler PG, Patterson RS. 1992.

Volatile effects of insect growth regulators against the

German cockroach (Dictyoptera: Blattellidae). J. Med.

Entomol., 29(2):364-367.

Bakudie E. 2006. Susceptibility of Tribolium castaneum

to novaluron on maize and rice. Bachelor of Science

Dissertation. Department of Crop Science, University of

Ghana, Legon, 36.

Boxall RA. 1986. A critical review of the methodology

for assessing farm grain losses after harvest. Tropical

Development and Research Institute Report G191, Viii

139.

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malathion and fenthion to dermestid larvae as influenced

by various surfaces. J. Econ. Entomol., 59(2): 253-254.

[CABI]. 2006. CAB International Data sheet for

Cadra cautella. Crop Protection Compendium 2006

Edition.

Chadwick PR. 1985. Surfaces and other factors

modifying the effectiveness of pyrethroids against

insects in public health. Pesticide Science 16(4):383-391.

Cutler CG, Tolman JH, Scott-Dupree CD and Harris

CR. 2005. Resistance potential of Colorado potato beetle

(Coleoptera: Crysomelidae) to Novaluron. J. Econ.

Entomol., 98(5):1685-1693.

Environmental Protection Agency (EPA). 2006.

Pesticides for horticulture production. Reference Guide,

27.

Eziah VY, Sacky I, Boateng BA, Obeng-Ofori D.

2011. Bioefficacy of neem oil (Calneem™), a botanical

insecticide against the tropical warehouse moth,

Ephestia cautella. Int. Res. J. Agric. Sci. Soil Sci.,

1(7): 242-248.

Ibrahim F. 2008. Effect of Rimon® 10EC (Novaluron)

on cocoa moth (Ephestia cautella) infesting stored cocoa

beans. Bachelor of Science Dissertation. Department of

Crop Science, University of Ghana, Legon, 33.

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Ishaaya, I., Kontsedalov, S., Mazirov, D., Horowitz,

AR. 2001. Biorational agents: mechanisms and

importance in IPM and IRM programs for controlling

agricultural pests. Med. Fac. Landbouww. Univ. Gent.

66, 363–374.

Kostyukovsky M, Trostanetsky A, Carmi Y, Frandji

H, Schneider R. 2003. Activity of novaluron on the

main stored product insects. In: Credland PF, Armitage

DM, Bell CH, Cogan PM and Highley E. [eds.],

Advances in Stored Product Protection. Proceedings of

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product protection. 22-26 July, York, UK, CABI

International, Walling ford, Oxon, 583-587.

Kostyukovsky M, Trostanetsky A. 2006. The effect of

a new chitin synthesis inhibitor, novaluron on various

developmental stages of Tribolium castaneum (Herbst).

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Lawes Agricultural Trust, 2007. Genstat 9.2. VSN

International Ltd., Hemel Hempstead, United Kingdom.

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Biology

A Checklist of Butterflies of Meenachil River Basin, Kerala, India

Keywords: Meenachil river, Endemic species, bio-indicators, anthropogenic pressure.

ABSTRACT:

Butterflies are highly sensitive to environmental change and are delicate creatures that act as good bio-indicators of the health of an ecosystem. Meenachil river basin has attracted considerable amount of public interest. A survey of the butterflies conducted randomly revealed a total of 91 species belonging to five families including three endemic species. Family Nymphalidae dominated in the study area, followed by Hesperiidae and Lycaenidae. This area is currently under severe anthropogenic pressure and minimizing these disturbances is important for the long-term survival of specialist butterflies.

768-774 | JRB | 2013 | Vol 3 | No 1





Research Journal

Authors:

Vincy MV1, Brilliant R2

and Pradeepkumar AP3

Institution: 1. School of Environmental

Science, Mahatma Gandhi

University, Kottayam,

Kerala.

2. PG Department of

Environmental Sciences,

St. John’s College, Anchal,

Kerala.

3. Department of Geology,

University of Kerala,

Kariavattom, Kerala.


Vincy MV.

Email:

[email protected]

Web Address: http://jresearchbiology.com/documents/RA0308.pdf.

Dates: Received: 21 Nov 2012 Accepted: 03 Dec 2012 Published: 04 Feb 2013

Article Citation: Vincy MV, Brilliant R and Pradeepkumar AP. A Checklist of Butterflies of Meenachil River Basin, Kerala, India. Journal of Research in Biology (2013) 3(1): 768-774


Original Research

INTRODUCTION

Butterflies are the most beautiful and colourful

creatures on the earth and have a great aesthetic value.

India harbours about 1501 species of butterflies

(Haribal, 1992), 285 species are found in southern India

(Thomas, 1966), of which 45 species are endemic to

southern India. Butterflies, widely appreciated for the

aesthetic value are important as ecological indicators

(Chakravarthy et al., 1997) and ‘flagship taxa” in

biodiversity inventories (Lawton et al., 1998).

Meenachil river which is one of the important

river of Kottayam district in Kerala, emerges from

Western Ghats and confluences into Vembanad Lake.

This river has a total length of 78 km and has

a catchment area of 1272 km2. The entire Meenachil

watershed area geographically lies between 9°25’ N

to 9°55’ N latitude and 76°30’ E to 77°00’ E longitude.

The general elevation of the entire river basin ranges

from 77 m to 1156 m in the high lands and less than 2 m

in the low lands. The Meenachil river basin falls within

the realm of tropical climate. The temperature of the area

varies in between 24°C and 32°C throughout the year.

The annual rainfall varies from less than 100 cm to more

than 500 cm with an average of 300 cm. The occasional

rainfall is also received between the two seasons. Rubber

trees are extensively cultivated in vast areas in the entire

river basin. Besides rubber, other crops like spices,

paddies etc., are also cultivated in the river basin area

(Watershed Atlas, 1996).

Among insects, butterflies are the most

studied group. Larsen (1987a, b, c, 1988) made a detailed

survey of butterflies of Nilgiri Mountains and recorded

nearly 300 species including endemics.

In Kerala, documentation of butterflies on Silent Valley

National Park (Mathew and Rahamathulla, 1993)

an d Pa r a m bi ku l a m Wi ld l i f e S an c tu a r y

(Sudeendrakumar et al., 2000) have been carried out.

The present paper presents a checklist and diversity of

butterfly populations in different altitude levels in

Meenachil river basin in Kerala, South India. However,

comprehensive long-term ecological studies to monitor

the butterfly population of the area remains as a serious

lacuna. Such studies are imperative to improve the

ecological utility of butterflies as indicator taxa.


The present study is an attempt to provide a

checklist of butterflies based on a four-year field study

from October 2008 to October 2012. Identification of

species was done using available literature (Evans, 1932;

Gunathilagaraj et al., 1998; Haribal, 1992; Palot et al.,

2003; Gay et al., 1992; Wynter-Blyth, 1957) and with

the help of experts. Species classification and scientific

names are as per Gunathilagaraj et al., (1998).

RESULT AND DISCUSSION

The study during the period indicate that the

habitats where butterflies were found and captured are

disturbed areas and are strongly influenced by

anthropogenic activities. These range from city lots to

pasture, abandoned fields, road sides, plantations,

riparian area, etc.

A total of 91 species belonging to 71 genera

distributed over five families were collected from the

monitoring sites, during the study period. The family

Nymphalidae dominated with 34 species followed

by Hesperiidae (20 spp.), Lycaenidae (18 spp.), Pieridae

(7 spp.), and Papilionidae (12 spp.). Even though, the

family Nymphalidae exhibited the maximum species

diversity, family Pieridae showed maximum species

density. Three butterfly species recorded from this region

have protected status under the Wildlife Protection Act,

1972 (Arora, 2003). They are Hypolimnas misippus and

Atrophaneura hector included under Schedule I Part IV

and one species Aeromachus pygmaeus in Schedule II

Part II. Further research with reference to ecology,

threats and conservation of butterflies in the area is in

progress.

Vincy et al., 2013


Vincy et al., 2013


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iae

(Fab

rici

us)

80-1

10 m

m

Com

mon

T

ho

ttea

sil

iqu

osa

9

Mal

abar

Rose

A

tro

phan

eura

pa

nd

iya

na (

Moore

) 100-1

30 m

m

Com

mon

T

ho

ttea

sil

iqu

osa

10

Cri

mso

n R

ose

*

Atr

op

han

eura

hec

tor

(Lin

nae

us)

90-1

10 m

m

Com

mon

A

rist

olo

chia

in

dic

a, T

ho

ttea

sil

iquo

sa

11

Com

mon

Bir

dw

ing

T

roid

es h

elen

a

(Lin

nae

us)

140-1

70 m

m

Not

rare

12

Sou

ther

n B

ird

win

g

Tro

ides

min

os

(Cra

mer

) 140-1

90 m

m

Not

rare

A

rist

olo

chia

in

dic

a, T

ho

ttea

sil

iquo

sa

FA

MIL

Y P

IER

IDA

E

13

Com

mon

Gra

ss Y

ello

w

Eu

rem

a h

eca

be

(Lin

nae

us)

40-5

0 m

m

Com

mon

C

aes

alp

inia

sp

p., C

ass

ia t

ora

, C

. fi

stu

la,

Aca

cia s

pp.

14

Com

mon

Em

igra

nt

Cato

psi

lia p

om

on

a (

Fab

rici

us)

55-8

0 m

m

Com

mon

B

au

hin

ia r

ace

mo

sa, C

. fi

stu

la, C

. to

ra,

Bu

tea

mon

osp

erm

a

15

Mott

led

Em

igra

nt

Cato

psi

lia p

yra

nth

e (L

inn

aeu

s)

50-7

0 m

m

Com

mon

C

. fi

stu

la, C

. to

ra

16

Gre

at O

ran

ge

Tip

H

ebo

mo

ia g

lau

cip

pe

(Lin

nae

us)

80-1

00 m

m

Com

mon

C

ap

pari

s sp

p.

17

Ch

oco

late

Alb

atro

ss

Ap

pia

s ly

nci

da (

Cra

mer

) 55-7

0 m

m

Com

mon

C

ap

pari

s sp

p.

18

Com

mon

Jez

ebel

D

elia

s eu

cha

ris

(Dru

ry)

66-8

3 m

m

Com

mon

D

end

rop

hth

oe

falc

ata

19

Psy

che

Lep

tosi

a n

ina (

Fab

rici

us)

35-5

0 m

m

Com

mon

C

leo

me

visc

osa

FA

MIL

Y N

YM

PH

AL

IDA

E

20

Blu

e T

iger

T

iru

ma

la l

imnia

ce (

Cra

mer

) 90-1

00 m

m

Com

mon

L

an

tan

a c

am

ara

, A

ger

atu

m c

on

yzoid

es,

Cro

tala

ria

ret

usa

21

Dar

k B

lue

Tig

er

Tir

um

ala

sep

ten

trio

nis

(B

utl

er)

75-9

5 m

m

Com

mon

A

ger

atu

m c

onyz

oid

es

22

Str

ipp

ed T

iger

D

an

au

s gen

uti

a (

Cra

mer

) 72-1

00 m

m

Com

mon

T

rid

ax

pro

cum

ben

s, L

anta

na s

pp

.,

Cro

tala

ria

ret

usa

23

Pla

in T

iger

D

an

au

s ch

rysi

ppu

s (L

inn

aeu

s)

70-8

0 m

m

Com

mon

C

alo

trop

is s

pp., A

ger

atu

m c

onyz

oid

es,

Tri

da

x p

rocu

mben

s,C

rota

lari

a r

etu

sa,

24

Gla

ssy

Blu

e T

iger

P

ara

nti

ca a

gle

a (

Sto

ll)

70-8

5 m

m

Com

mon

C

alo

trop

is s

pp.,

Ag

eratu

m c

onyz

oid

es,

Sta

chyt

arp

het

a s

pp., C

rota

lari

a r

etu

sa

Tab

le 1

. L

ist

of

bu

tterfl

y s

pec

ies

coll

ecte

d i

n t

he

stu

dy

area

s a

nd

th

e p

lan

ts v

isit

ed

by

th

em

Vincy et al., 2013


25

Com

mon

Cro

w

Eu

plo

ea c

ore

(C

ram

er)

85-9

5 m

m

Com

mon

Ic

hn

oca

rpu

s fr

ute

scen

s, H

emid

esm

us

ind

icu

s, F

icu

s sp

p., S

treb

lus

asp

er,

Ag

eratu

m c

onyz

oid

es, C

rota

lari

a s

pp.,

Chro

mo

laen

a o

dora

ta

26

Com

mon

Naw

ab

Po

lyu

ra a

tha

ma

s (D

rury

) 60-7

5 m

m

Com

mon

A

caci

a p

enn

ata

, A

den

an

ther

a p

avo

nin

a

27

Com

mon

Even

ing

Bro

wn

Mel

an

itis

led

a (

Lin

nae

us)

60-8

0 m

m

Com

mon

O

ryza

sa

tiva

, P

anic

um

sp

p.

28

Bam

bo

o T

reeb

row

n

Let

he

euro

pa (

Fab

rici

us)

65-7

5 m

m

Com

mon

B

am

bu

sa s

pp

.

29

Com

mon

Pal

mfl

y

Ely

mn

ias

hyp

erm

nes

tra (

Lin

nae

us)

60-8

0 m

m

Com

mon

A

reca

ca

tech

u, C

oco

s nu

cife

ra

30

Com

mon

Bu

shbro

wn

M

ycale

sis

per

seu

s (F

abri

ciu

s)

38-5

5 m

m

Com

mon

O

ryza

sp

p.

31

smooth

-eye

d b

ush

bro

wn

O

rso

tria

ena

med

us

(Fab

rici

us)

45-5

5 m

m

Com

mon

O

ryza

sa

tiva

32

Com

mon

Fiv

erin

g

Yp

thim

a b

ald

us

(Fab

rici

us)

32-4

8 m

m

Com

mon

33

Com

mon

Fourr

ing

Y

pth

ima h

ueb

ner

i (K

irb

y)

30-4

0 m

m

Com

mon

G

rass

es

34

Taw

ny

Cost

er

Acr

aea

vio

lae

(Fab

rici

us)

50-6

5 m

m

Com

mon

A

po

rosa

lin

dle

yana

, P

ass

iflo

ra f

oet

ida

35

Tam

il Y

eom

an

Cir

roch

roa th

ais

(F

abri

ciu

s)

60-7

5 m

m

Com

mon

H

ydn

oca

rpu

s sp

p.

36

Ru

stic

C

up

ha e

rym

an

this

(D

rury

) 50-6

0 m

m

Com

mon

37

Com

mon

Leo

par

d

Ph

ala

nta

ph

ala

nth

a (

Dru

ry)

50-6

0 m

m

Com

mon

38

Com

man

der

M

odu

za p

rocr

is (

Cra

mer

) 60-7

5 m

m

Com

mon

O

chre

ina

ucl

ea m

issi

on

is,

Mu

ssa

end

a

fro

ndo

sa

39

Com

mon

Las

car

Pa

nto

pori

a h

ord

onia

(S

toll

) 45-5

0 m

m

Com

mon

A

caci

a p

enn

ata

40

Com

mon

Sai

lor

Nep

tis

hyl

as

(Lin

nae

us)

50-6

0 m

m

Com

mon

D

alb

ergia

sp

p.,

Ziz

yph

us

spp

., T

hes

pes

ia

po

pu

lnea

, G

rew

ia s

pp

., B

om

ba

x a

lab

ari

cum

41

Cli

pp

er

Pa

rth

eno

s sy

lvia

(C

ram

er)

95-1

30 m

m

Com

mon

ti

no

spo

ra c

ord

ifo

lia

42

Com

mon

Bar

on

Eu

thali

a a

con

thea

(H

ewit

son

)

55-8

0 m

m

Com

mon

A

na

card

ium

occ

iden

tali

s, M

angif

era

in

dic

a,

Str

eblu

s a

sper

43

Gre

y C

ou

nt

Ta

naec

ia l

epid

ea (

Bu

tler

) 65-8

5 m

m

Com

mon

C

are

ya a

rbo

rea

44

Com

mon

Map

C

yres

tis

thyo

da

ma

s (B

ois

du

val

) 50-6

0 m

m

Not

Com

mon

F

icu

s sp

p.

45

Pai

nte

d L

ady

Va

nes

sa c

ard

ui

(Lin

nae

us)

55-7

0 m

m

Com

mon

B

lum

ea s

pp.

46

An

gle

d C

aste

r A

ria

dne

ari

ad

ne

(Lin

nae

us)

45-6

0m

m

Un

com

mon

R

icin

us

com

mu

nis

47

Com

mon

Cas

ter

Ari

ad

ne

mer

ione

(Cra

mer

) 45-6

0m

m

Com

mon

R

icin

us

com

mu

nis

48

Ch

oco

late

Pan

sy

Jun

onia

ip

hit

a (

Cra

mer

) 55-8

0 m

m

Com

mon

49

Gre

y P

ansy

Ju

nonia

atl

ites

(L

inn

aeu

s)

55-6

5 m

m

Com

mon

50

Pea

cock

Pan

sy

Jun

onia

alm

an

a (

Lin

nae

us)

60-6

5 m

m

Com

mon

O

sbec

kia s

pp

.

51

Lem

on

Pan

sy

Jun

onia

lem

on

ias

(Lin

nae

us)

40-6

0 m

m

Com

mon

S

ida r

ho

mb

ifo

lia

52

Gre

at E

gg

fly

Hyp

oli

mn

as

bo

lin

a (

Lin

nae

us)

70-1

10 m

m

Com

mon

S

ida r

ho

mb

ifo

lia

, H

ibis

cus

spp

.

53

Dan

aid

Eg

gfl

y*

Hyp

oli

mn

as

mis

ipp

us

(Lin

nae

us)

70-8

5 m

m

Com

mon

H

ibis

cus

spp

.

FA

MIL

Y L

YC

AE

NID

AE

54

Ape

Fly

S

palg

is e

piu

s (W

estw

ood

) 20-3

0 m

m

Not

com

mon

C

arn

ivo

rou

s ca

terp

illa

rs f

eed

on

mea

ly b

ugs

55

India

n S

un

bea

m

Cure

tis

thet

is (

Hü

bn

er)

40-4

8 m

m

Not

rare

A

bru

s p

reca

tori

us,

Po

nga

mia

pin

na

ta

56

Red

Sp

ot

Zes

ius

chry

som

all

us

(Hü

bn

er)

38-4

4 m

m

Not

rare

C

ate

rpil

lars

fee

d o

n a

nt

larv

ae

57

Yam

fly

Lo

xura

aty

mnu

s (C

ram

er)

36-4

0 m

m

Com

mon

S

mil

ax

spp., D

iosc

ore

a p

enta

phyl

la

58

Mon

key

Pu

zzle

R

ath

ind

a a

mo

r (F

abri

ciu

s)

26-2

8 m

m

Not

rare

Ix

ora

sp

p.

Vincy et al., 2013


59

Sla

te F

lash

R

ap

ala

ma

nea

(H

ewit

son

) 30-3

3 m

m

Com

mon

A

caci

a p

enn

ata

60

Com

mon

Sil

ver

lin

e C

iga

riti

s v

ulc

an

us

(Fab

rici

us)

26-3

4 m

m

Com

mon

Z

izyp

hus

rug

osa

, C

anth

ium

oro

ma

nd

elic

um

,

Cle

roden

dru

m i

ner

me

61

An

gle

d P

ierr

ot

Cale

ta c

ale

ta (

Hew

itso

n)

26-3

2 m

m

Not

rare

Z

izyp

hus

rug

osa

62

Ban

ded

Blu

e P

ierr

ot

Dis

cola

mpa

eth

ion (

Cra

mer

) 26-3

0 m

m

Com

mon

Z

izip

hus

spp.

63

Com

mon

Pie

rrot

Cast

ali

us

rosi

mon

(F

abri

ciu

s)

24-3

4 m

m

Com

mon

Z

izyp

hus

rug

osa

, Z

. ju

jub

a

64

Com

mon

Lin

e B

lue

Pro

sota

s n

ora

(C

. F

eld

er)

18-2

5 m

m

Com

mon

A

caci

a c

ate

chu, M

imo

sa s

pp.

65

Dar

k C

erule

an

Jam

ides

boch

us

(Sto

ll)

25-3

4 m

m

Com

mon

B

ute

a m

on

osp

erm

a, C

rota

lari

a s

pp

.,

Po

nga

mia

pin

nata

66

Com

mon

Cer

ule

an

Jam

ides

cel

eno (

Cra

mer

) 27-4

0 m

m

Com

mon

B

ute

a m

on

osp

erm

a,

Po

nga

mia

pin

na

ta,

Ab

rus

pre

cato

riu

s

67

Forg

et m

e n

ot

Cato

chry

sop

s st

rab

o (

Fab

rici

us)

25-3

5 m

m

Com

mon

D

esm

od

ium

sp

p.

68

Les

ser

Gra

ss B

lue

Ziz

ina

oti

s (F

abri

ciu

s)

19-2

6 m

m

Com

mon

F

ab

ace

ae

spp.

69

Red

Pie

rrot

Ta

lica

da n

yseu

s (G

uér

in)

30-3

6 m

m

Com

mon

70

Qu

aker

N

eop

ithec

op

s za

lmo

ra (

Bu

tler

) 16-3

0 m

m

Com

mon

G

lyco

smis

pen

tap

hyl

la

71

Plu

m J

ud

y A

bis

ara

ech

eriu

s (S

toll

) 40-5

0 m

m

Com

mon

FA

MIL

Y H

ES

PE

RII

DA

E

72

Com

mon

Sp

ott

ed F

lat

Cel

aen

orr

hin

us

leuco

cera

(K

oll

ar)

45-5

5 m

m

Com

mon

73

India

n S

kip

per

S

pia

lia g

alb

a (

Fab

rici

us)

20-2

7 m

m

Com

mon

S

ida r

ho

mb

ifo

lia

, H

ibis

cus

spp

.

74

Com

mon

Sm

all

Fla

t S

ara

nges

a d

asa

hara

(M

oore

) 26-3

5 m

m

Com

mon

A

syst

asi

a s

pp.

75

Tri

colo

ur

Pie

d F

lat

Cola

den

ia i

nd

ran

i (M

oore

) 40-4

6 m

m

Com

mon

M

all

otu

s p

hil

ippin

ensi

s, D

esm

odiu

m s

pp

.

76

Su

ffu

sed

Sn

ow

Fla

t T

ag

iad

es g

ana (

Moore

) 45-5

0 m

m

Not

rare

77

Wat

er S

now

Fla

t T

ag

iad

es l

itig

iosa

(M

ösc

hle

r)

37-4

4 m

m

Not

rare

S

mil

ax

spp.

78

Tam

il G

rass

Dar

t T

ara

ctro

cera

cer

am

as

23-3

0 m

m

Com

mon

O

ryza

sa

tiva

, g

rass

es

79

Com

mon

Gra

ss D

art

Ta

ract

roce

ra m

aev

ius

(Fab

rici

us)

22-2

8 m

m

Com

mon

G

rass

es

80

Com

mon

Dar

tlet

O

rien

s go

loid

es (

Moore

) 24-2

8 m

m

Com

mon

81

Dar

k P

alm

Dar

t T

elic

ota

anci

lla (

Her

rric

h-S

chäf

fer)

33-3

6 m

m

Com

mon

C

oco

s n

uci

fera

, O

ryza

sp

p., S

acc

ha

rum

sp

p.

82

Ric

e S

wif

t B

orb

o c

inn

ara

(W

alla

ce)

30-3

6 m

m

Com

mon

O

ryza

sa

tiva

, P

enn

iset

um

sp

p., I

sch

aem

um

spp

., C

ymb

opo

go

n s

pp.

83

Con

tig

ou

s S

wif

t P

oly

trem

is l

ub

rica

ns

(Her

rric

h-

Sch

äffe

r)

36-4

2 m

m

Not

com

mon

84

India

n P

alm

Bob

Su

ast

us

gre

miu

s (F

abri

ciu

s)

32-4

5 m

m

Com

mon

C

ala

mu

s sp

p., C

ary

ota

ure

ns,

Coco

s n

uci

fera

85

Gai

nt

Red

eye

G

an

ga

ra t

hyr

sis

Fab

rici

us)

70-7

6 m

m

Not

rare

C

ala

mu

s ro

tan

g,

Ca

ryo

ta u

ren

s, C

oco

s n

uci

fera

86

Com

mon

Red

eye

Ma

tap

a a

ria

(M

oore

) 40-5

5 m

m

Com

mon

87

Ch

estn

ut

Bob

Iam

bri

x sa

lsa

la (

Moore

) 26-3

0 m

m

Com

mon

G

rass

es a

nd b

am

bo

os

88

Res

tric

ted

Dem

on

N

oto

cryp

ta c

urv

ifa

scia

(F

eld

er &

Fel

der

)

38-5

0 m

m

Com

mon

C

ost

us

spec

iosu

s

89

Gra

ss D

emon

U

da

spu

s fo

lus

(Cra

mer

) 40-4

8 m

m

Com

mon

Z

ing

iber

sp

p.

90

Pyg

my

Scr

ub H

op

per

**

A

ero

mach

us

pyg

maeu

s (F

abri

ciu

s)

20-2

2 m

m

Com

mon

91

India

n A

ce

Ha

lpe

hom

ole

a (

Hew

itso

n)

30-3

6 m

m

Com

mon

B

am

bo

o

* -

in

dic

ate

s sp

eci

es

com

ing

un

der S

ch

ed

ule

I P

art

IV a

nd

** -

Sch

ed

ule

II

Pa

rt I

I of

Th

e W

ild

life

(P

rote

cti

on

) A

ct,

19

72

The study shows that the sustained interference

and disturbance seem to affect the occurrence and

numerical strength of each butterfly species. If this

situation goes unabated, the abundant butterflies may

become rare and the less abundant ones could disappear

permanently. Further, the decline in the number of

butterflies largely allows inbreeding which becomes fatal

in course of time. Modified habitats with reduced plant

cover contribute to warm conditions and these conditions

might allow some butterflies to extend their distribution

to different habitats. The butterflies which control certain

plant pets, if decline in number or disappear from the

habitat, plants too get affected because of the unchecked

plant pets. Therefore, the very presence of butterflies in

species and number may be taken as an indication of the

health of the habitat.

REFERENCES

Arora K. 2003. Forest Laws. The Wildlife Protection

Act, 1972 as amended by the Wildlife (Protection)

Amendment Act, 2002 (Act 16 of 2003). Published by

Professional Book Publishers, New Delhi, 85.

Chakravarthy AK Rajagopal D and Jagannatha R.

1997. Insects as bio indicators of conservation in the

tropics. Zoos’ Print, 12:21-25.

Evans WH. 1932. Identification of Indian Butterflies.

Bombay Natural History Society, Bombay, 454.

Gay T Kehimkar ID and Punetha JC. 1992. Common

Butterflies of India. Oxford University Press, Bombay.

Gunathilagaraj K. 1998. Some South Indian Butterflies.

Tamil Nadu, India: Nilgiri Wildlife and Environments

Association, Udhagamandalam, Nilgiris, 274.

Haribal M. 1992. The butterflies of Sikkim, Himalaya

and their natural history. Nataraj Publishers, Dehradun,

217.

Mathew G and Rahamathulla VK. 1993. Studies on

the butterflies of Silent Valley National Park. Entomon,

18:185-192.

Larsen TB. 1987a. The butterflies of the Nilgiri

mountains of South India (Lepidoptera: Rhopalocera).

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43.

Larsen TB. 1987b. The butterflies of the Nilgiri



316.

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584.

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43.

Lawton, JH Bignell D E Bolton B Bloemers GF

Eggleton P Hammond PM Hodda M Holts RD

Larsen TB Mawdsley NA Stork NE Srivastava DS

and Watt AD. 1998. Biodiversity inventories indicator

taxa and effect of habitat modification in tropical forest.

Nature, 391: 72-76.

Palot J Balakrishnan VC and Kambrath B. 2003.

Keralathile Chitrasalabhangal. Malabar Natural History

Society, Calicut, Kerala, 195.

Sudheendrakumar VV Binoy CF Suresh PV and

Mathew G. 2000. Habitat associations of butterflies in

the Parambikulam Wildlife Sanctuary, Kerala, India.


201.

Vincy et al., 2013


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Thomas S. 1966. Bulletin of the Madras Government

Museum - Descriptive Catalog of the Butterflies, Natural

History Section Vol. VII No. 1.

Watershed Atlas. 1996. Kerala State Land Use board,

Govt. of Kerala Publications, Kerala.

Wynter-Blyth MA. 1957. Butterflies of Indian Region.

Bombay Natural History Society, Bombay.


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Biology

Microbial production of glutaminase enzyme

Keywords: Actinomycetes, Anticancer properties, Enzymes, Glutamic acid and L-Glutaminase.

ABSTRACT: Enzymes are proteins highly specific in their actions on substrates and serve as biocatalysts. They are produced by cells in order to accelerate both the rate and specificity of metabolic reactions. Microbial enzymes are known for their unique characteristics over other sources due to their easy production on a commercial scale and stability. Different microorganisms are known to produce various enzymes such as bacteria, fungi and actinomycetes which produce a variety of extra-cellular and endo-cellular enzymes. Some of these actinomycetes enzymes have been isolated from the culture filtrates or the mycelium, concentrated and purified. Others have only been demonstrated in the mycelium of the organism. However, the ability to produce a variety of enzymes may be an attractive phenomenon in these microorganisms since they are nutritionally quite versatile. Microbial L-glutaminase has recently gained more attention due to its anticancer properties, in addition to its use as a flavor enhancer in food industry by increasing the amount of glutamic acid content in the fermented food .

775-779 | JRB | 2013 | Vol 3 | No 1






Research Journal

Authors:

Mario Khalil Habeeb

Institution:

Microbiology Department,

Faculty of Science, Ain

Shams University, 15566

El-Khalifa El-Mamoun

street, Abbassia, Cairo,

Egypt, Postal code: 11566.


Mario Khalil Habeeb.

Email:

[email protected],

[email protected]

Telephone:

+20 (02) 22409635

Mobile:

+20 (0128) 3941815


Dates: Received: 16 Jan 2013 Accepted: 22 Jan 2013 Published: 06 Feb 2013

Article Citation: Mario Khalil Habeeb. Microbial production of glutaminase enzyme. Journal of Research in Biology (2013) 3(1): 775-779


OVER VIEW

INTRODUCTION

Enzymes are highly selective catalytic proteins

produced by living cells which may or may not contain a

non-protein prosthetic group (Underkofler et al., 1958).

Actinomycetes are considered to be preferred

enzymes sources due to their production of extracellular

enzymes. They are highly diverse group with numerous

members representing important source of microbial

enzymes. Actinomycetes genera are differentiated from

each other based on morphological, biochemical, and

physiological criteria. They act as decomposers of

complex animal and plant materials resulting in release

of simple substances, especially carbon and nitrogen

which is easily utilized by other organisms, thus

performing a vital role in life cycle. Due to their

significant biochemical activities, Actinomycetes are

used in commercial production of various substances

such as antibiotics and enzymes (Waksman, 1950).

Because of its industrial and pharmaceutical

applications, intensive research was conducted on

L-glutaminase recently. L-glutaminase is produced by

various terrestrial microorganisms such as Pseudomonas

sp., Acinetobacter sp., Escherichia coli, Bacillus sp.,

Hansenula sp., Candida sp., Aspergillus oryzae and

Beauveria bassiana (Sabu, 2003). Also few

marine microorganisms such as Micrococcus luteus,

Vibrio cholera and Pseudomonas fluorescens were

reported to produce the enzyme (Chandrasekaran, 1997).

Definition

L-glutaminase is classified as an amidohydrolase

enzyme which acts upon amide bonds of L-glutamine

generating L-glutamic acid and ammonia. It is present

in both microorganisms and mammalian tissues

(Ohshima et al., 1976b). Microbial sources of

glutaminase showed a great role in various applications

such as its use in fermented foods precisely in soy sauce

and other related types, in addition to its use as

anticancer agent which act by inhibition of glutamine

utilization by the cancerous cells resulting in selective

starvation of cancerous cells and their possible death

(Santana et al., 1968).

Glutaminase Producing microorganisms

Different types of organisms were reported

to produce glutaminase enzyme. However, The selection

of the right organism is very critical to obtain high yield

of the required enzyme (Akujobi et al., 2012).

L-glutaminases from E. coli, Pseudomonas sp.,

Bacillus sp., and Clostridium welchii have been isolated

and well studied (Wade et al., 1971). In addition to these

bacterial sources, the fungus Aspergillus oryzae showed

a great ability to produce this enzyme. Among

microorganisms, actinomycetes are widely recognized

as preferable L-glutaminase sources because they

generally produce extracellular enzymes, which facilitate

the enzyme recovery from the fermentation broth

such as glutaminase from Streptomyces rimosus

(Sivakumar et al., 2006).

Microbial Glutaminase Characteristics

Temperature is considered to be an important

factor affecting the enzyme stability, The optimum

temperature recorded by many glutaminases ranged from

40-50ºC. However, the temperature stability of

glutaminase I (Micrococcus glutaminase) of M. luteus

could be increased by the addition of 10% NaCl

(Moriguchi et al., 1994). The optimum temperature for

A. oryzae glutaminase was around 37-45ºC and remained

stable at up to 45ºC and the enzyme was completely

inactive at 55ºC (Nakadai and Nasuno, 1989).

It is interesting that the exposure of E. coli

glutaminase B to cold resulted in a reversible

inactivation of enzymatic activity, while subsequent

warming to 24ºC restored the activity. There was no

difference in the molecular weight of the cold inactivated

enzyme and the warm activated enzyme. The

conformational changes which probably occur upon

exposure to cold resulted from a weakening of the

interaction among hydrophobic groups in the protein

(Chou et al., 1993).

Habeeb., 2013


The salt-tolerance of glutaminase is an important

parameter in industrial processes that include

high-salinity. It was reported that the high-salt

concentration (nearly 3 M NaCl) used in the process

of soy sauce fermentation resulted in remarkable

inhibition of the koji mold (A. oryzae) Glutaminase

(Koibuchi et al., 2000).

Methods Used for Microbial Glutaminase Production

Two methods are known for the production of

microbial glutaminase.

Submerged (Liquid) Production Method

In this method, the sterile media together with

the enzyme producing organism were introduced into

large fermentors (Tanks) followed by constant mixing

and supply of sterile air (Schuegerl et al., 1991).

Actinomycetes glutaminases showed a high salt

tolerance in this production method. Reports showed that

Streptomyces rimosus isolated from estuarine fish

recorded high salt tolerance and the highest enzyme

production obtained at temperature 27ºC, pH 9.0 and

both glucose and malt extract proved to be the best

carbon and nitrogen sources for maximum enzyme

production (Imada et al., 1973).

Surface Production Method

This method includes the use of solid support on

which microorganisms are grown. Surface production

method (solid state fermentation) showed 25 to 30 fold

increase in enzyme production when compared with

submerged production (Sabu et al., 2000b).

Wheat bran was found to be a favorable support

for microorganisms in the process of glutaminase

production (Kashyap et al., 2002). In addition to wheat

bran many other solid supports showed high efficiency in

the enzyme production such as ground nut cake powder,

copra cake powder and sesamum oil cake (Prabhu and

Chandrasekaran, 1995). Polystyrene beads, supported by

mineral salts and glutamine are another form of solid

supports used for the enzyme production.

By using this method it was found that L-glutaminase

producing marine alkalophilic Streptomyces sp. SBU1

which was isolated from Cape Comorin coast,

India gave highest enzyme production after 4 days

of incubation and at 14% Corn steep liquor

(Krishnakumar et al., 2011).

Applications

L-glutaminase has received great attention due to

its valuable applications in several fields especially in

medicine and its use as an anticancer agent either alone

or together with any other agents is known as enzyme

therapy, In addition to its role as flavor enhancer by

increasing the glutamic acid content of food. Also

glutaminase applications extend to the enzyme utilization

as biosensor in analytical purposes by measuring the

levels of L-glutamine and finally in the manufacture of

fine chemicals such as theanine when used with baker’s

yeast.

Glutaminase as Enzyme Therapy

Glutaminase can be used as alternative for cancer

treatment as enzyme therapy. The mechanism for

glutaminase therapy includes that L-Glutaminase act on

its substrate (L-glutamine) and breaks it down leading to

the selective destruction of the tumor cells accompanied

by inhibition of both protein and nucleic acid

biosynthesis due to glutamine starvation and this is

attributed to the inability of cancerous cells to synthesis

glutamine (Tanaka et al., 1988). This is due to the fact

that some types of cancerous cells utilize glutamine

greatly (Lazarus and Panasci, 1986). Concerning this

finding various enzymatic therapies developed to deprive

L-glutamine to cancerous cells (Roberts et al., 1970).

Glutaminase as Flavor Enhancer

Glutamate is a famous amino acid and

considered as a natural constituent of many fermented or

aged foods, such as soy sauce, fermented bean paste and

cheese (O`Mahony and Ishi, 1987). It gives these types

of food their desired taste (Chou and Hwan, 1994).

Glutamate (Glutamic acid) accumulated in these food

Habeeb., 2013


types as a result of protein hydrolysis by proteolytic

enzymes such as glutaminase and protease have a vital

role in food industry (Tambekar and Tambekar, 2011).

Glutaminase as biosensor

L-glutaminase is used as biosensor to monitor the

L-glutamine levels in body fluids. This technique is more

applicable than previously used methods and

characterized by its high specificity compared with cell

based sensors in addition to its fast response. This has led

to intensive use of glutaminase in clinical purposes

especially that is derived from mammalian tissues.

Glutaminase and Manufacture of Various Chemicals

Theanine (γ-l-glutamyl ethylamide) is

synthesized by theanine synthetase (EC 6.3.1.6) in plants

and known for its capability to inhibit stimulation by

caffeine, in order to enhance the effects of the anticancer

agents. Bacterial glutaminases together with baker’s

yeast are used to produce theanine (Tachiki et al., 1998).

Also L-glutaminase is used in the manufacture of

γ –glutamyl alkamides by the transfer of γ-glutamyl from

a donor molecule such as glutamine or glutathione to a

glutamyl acceptor like ethylamine or glycyl glycine by

catalysis.

Conclusion

Due to their important applications, Microbial

glutaminases gained much attention among the

commercially important enzymes. Their role in the

biotechnological industries, in addition to their medical

applications as anticancer agents created the need for

searching of high potential microorganisms strains. The

advantages of the microbial glutaminases - such as their

stability and large scale production - over other sources

made microorganisms represent a desirable source for

the enzyme production. This brief review revealed the

microbial sources of the enzyme and its characteristics,

in addition to the production methods and extended to its

various applications.

REFERENCES

Akujobi CO, Odu NN, Okorondu SI and

Ike GN. 2012. Production of protease by

Pseudomonas aeruginosa and Staphylococcus aureus

isolated from abattoir environment. Journal of Research

in Biology. 2(2):077-082

Chandrasekaran M. 1997. Industrial enzymes from

marine microorganisms. J Mar Biotech., 5:86-89.

Chou CC, Yu RC, Tsai CT. 1993.

Production of glutaminase by Actinomucor elegans,

Actinomucor taiwanensis and Aspergillus oryzae.

J Chinese Agric Chem Soc., 31:78-86.

Chou CC, Hwan CH. 1994. Effect of ethanol on the

hydrolysis of protein and lipid during the ageing of a

Chinese fermented soya bean curdsufu. J Sci Food

Agric., 66(3):393-398.

Imada A, Igarasi S, Nakahama K and Isono M. 1973.

Asparaginase and Glutaminase activities of

microorganisms. J Gen Microbiol., 76:85-99.

Kashyap P, Sabu A, Pandey A, Szakacs G and Soccol

CR. 2002. Extra-cellular L-glutaminase production by

Zygosaccharomyces rouxii under solid-state

fermentation. Process Biochem., 38(3):307-312.

Koibuchi K, Nagasaki H, Yuasa A, Kataoka J

and Kitamoto K. 2000. Molecular cloning and

characterization of a gene encoding glutaminase from

Aspergillus oryzae. Appl Microbiol Biotechnol., 54

(1):59-68.

Krishnakumar S, Alexis R, Rajan and Ravikumar S.

2011. Extracellular production of L-glutaminase by

marine alkalophilic Streptomyces sp. SBU1 isolated from

Cape Comorin coast. Ind J Geo-Marine Sci., 40(5):717-

721.

Lazarus P, Panasci LC. 1986; Characterization of

L-Threonine and L-glutamine transport in murine P388

leukaemia cells in vitro. Biochim Biophys Acta 856

(3):488-495.

Moriguchi M, Sakai K, Tateyama R, Furuta Y

and Wakayama M. 1994. Isolation and characterization

of salt-tolerant glutaminase from marine

Micrococcus luteus K-3. J Ferment Bioeng. 77(6):621-

625.

Habeeb., 2013


Nakadai T, Nasuno S. 1989. Use of glutaminase for soy

sauce made by Koji or a preparation of proteases from

Aspergillus oryzae. J Ferment Bioeng., 67(3):158-162.

Ohshima M, Yamamoto T and Soda K. 1976b.

Further characterization of glutaminase isozymes

from Pseudomonas aeruginosa. Agri Biologi Chem.

40(11):2251-2256.

O`Mahony M and Ishi M. 1987. The umami taste

concept: Implications for the dogma of four basic tastes

in Umami. Marcel Dekker, New York. 75-93.

Prabhu GN, Chandrasekaran M. 1995. Polystyrene -

an inert carrier for glutaminase production by marine

Vibrio costicola under Solid state fermentation. World J

Microbiol Biotechnol., 11(6):683-684.

Roberts J, Holcenberg JS and Dolowy WC. 1970.

Antineoplastic activity of highly purified bacterial

glutaminase. Nature 227:1136-1137.

Sabu A. 2003. Sources, properties and applications of

microbial therapeutic enzymes. Ind J Biotechnol., 2

(3):334-341.

Santana CF de, Pinto Kde V, Moreira LC and

Lacerda AL. 1968. Action of swine kidney

L-glutaminase on Ehrlich carcinoma. Rev Inst Antibiot. ;

8(1):105-107.

Schuegerl K, Brandes L, Dullau T, Holzhauer-Rieger

K, Hotop S and Huebner U. 1991. Fermentation

monitoring and control by on-line flow injection and

liquid chromatography. Anal Chim Acta. ; 249(1):87-

100.

Sivakumar K, Sahu MK, Manivel PR and Kannan L.

2006. Optimum conditions for L-glutaminase production

by actinomycete strain isolated from estuarine fish,

Chanos chanos. Ind J Exp Biol., 44(3):256-258.

Tachiki T, Yamada T, Mizuno K, Ueda M, Shiode J

and Fukami H. 1998. γ-Glutamyl transfer reactions

by glutaminase from Pseudomonas nitroreducens

IFO 12694 and their application for the syntheses of

theanine and γ-glutamylmethylamide. Biosci Biotechnol

Biochem., 62:1279-1283.

Tambekar DH and Tambekar SD. 2011.

Partial characterization and optimization of protease

production from newly isolated Cohnella thermotolerans

from Lonar Lake. Journal of Research in Biology.

1(4):292-298.

Tanaka S, Robinson EA, Appella E, Miller M,

Ammon HL, Roberts J, Weber IT and Wlodawer A.

1988. Structures of amidohydrolases. Amino acid

sequence of a glutaminase-asparaginase from

Acinetobacter glutaminasifrcans and preliminary

crystallographic data for an asparaginase from

Erwinia chrysanthemi. J Biol Chem., 263:8583-8591.

Underkofler LA, Barton RR and Rennert SS. 1958.

Production of microbial enzymes and their applications.

Appl Microbiol., 6(3):212-221.

Wade HE, Robinson HK and Phillips BW. 1971.

Asparaginase and glutaminase activities of bacteria.

J Gene Microbiol. 69:299-312.

Waksman SA. 1950. The actinomycetes: nature,

occurance and activities. Waverly press, Baltimore,

U.S.A.

Habeeb., 2013



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Biology

A review on the role of nutrients in development and

organization of periphyton

Keywords: Aquatic ecosystem, Biofilm, carbon, primary productivity, phytoplankton.

ABSTRACT:

Periphyton communities have not received wider attention and often misunderstood with ‘biofilm’ for their nature of development and role in aquatic ecosystem. To clarify its functional objective in aquatic ecosystem, present review proposes a functional definition for ‘periphyton’ in terms of ecological interactions and also outlines its ecological role in nutrient sharing with other aquatic components. The development and succession of periphyton is a function of nutrient and carbon (C) sharing with its constituent parts and ambient environment. Through mechanisms like entrapment, de novo synthesis, nutrient leakage, trophic upgrading etc., ambient nutrients are routed to periphyton and transferred to upper trophic levels. Periphyton communities stand next to phytoplankton for their contribution to primary productivity, in nutrient rich aquatic environment. Unlike phytoplankton, nutrient poor aquatic environment has no effect on periphytic primary productivity. As periphyton communities are attached to substratum, their ability to assimilate organic nutrient through substratum is an additional advantage over phytoplankton.

780-788 | JRB | 2013 | Vol 3 | No 1






Research Journal

Authors:

Saikia SK1, Nandi S2,

Majumder S3.

Institution:

1. Assistant Professor,

Aquatic Ecology Laboratory,

Department of Zoology,

Visva Bharati University,

Santiniketan, West Bengal,

India, Pin-731235.

2. Research Fellows,

Aquatic Ecology Laboratory, Department of Zoology,


Santiniketan, West Bengal,

India.

3. Research Fellows,

Aquatic Ecology Laboratory,

Department of Zoology,


Santiniketan, West Bengal.


Saikia SK.

Email: [email protected]

Web Address:

http://jresearchbiology.com/

documents/RA0307.pdf.

Dates: Received: 20 Nov 2012 Accepted: 10 Dec 2012 Published: 11 Feb 2013

Article Citation: Saikia SK, Nandi S, Majumder S. A review on the role of nutrients in development and organization of periphyton.

Journal of Research in Biology (2013) 3(1): 780-788


Original Research

INTRODUCTION

The term „periphyton‟ (peri round; phyton plant)

was proposed by (Behning, 1924) and popularized

by several authors (Cooke, 1956; Sladeckova, 1962;

Pieczynska, 1970). There exist a series of definitions

proposed for „periphyton‟ (Young, 1945; Neel, 1953;

Wetzel, 1963). (Wetzel, 1983) defined it as the micro

„floral‟ community living attached to any substrate

under water. (Stevenson, 1996) used it for describing

microorganisms such as algae and bacteria growing in

association with substrata. These communities play an

important role in water bodies, not only as important

primary producers and energy source for higher trophic

levels, but also by affecting the nutrient turnover and the

transfer of nutrients between the benthic and the pelagic

zone (review Saikia, 2011). The substrates of periphyton

commonly include submerged plants or plant parts,

rocks and sediments. Such substrate selection designs

periphyton as a medium in transferring and „trophic

upgrading‟ of nutrients. This property recognizes

periphyton as a tool for biofiltering excess nutrients from

polluted waters and for efficient nutrient transfer in

aquatic food chain.

Aquatic ecosystems mainly comprise of

freshwater and marine water bodies and their ecology

discusses the relationship of aquatic organisms and

its interaction with the immediate environment. The

principle biotic components primarily explained in the

recent past for their contributions to different interactions

in aquatic ecosystem are macrophytes, plankton

(Zooplankton and phytoplankton) and invertebrates

(benthos, nekton and neuston). Till the mid of

19th century, periphyton or „associated organisms‟ were

not given any biological credit for their role in aquatic

ecosystem. Probably, (Wetzel, 1963) in his evolutionary

review paper „Primary productivity of periphyton‟ in

Nature, for the first time made convincing remark on the

role of periphyton in aquatic ecosystem. Even today,

periphytic communities are ignored as a major

contributor of most of the nutrient inputs to aquatic

ecological cycles. The present review is an attempt to

outline periphyton as an integral and essential component

of aquatic ecosystem highlighting few areas recently

addressed on the role of periphyton in nutrient sharing in

aquatic processes.

Periphyton: A Nutrient Dependent Organization

(Whal, 1989) discussed settling pattern

of „biofilm‟ (Figure 1), in four phases: (i) surface

conditioning or adsorption of dissolved organic

compounds where macromolecules attach to submerged

surfaces following a spontaneous physical-chemical

process; (ii) primary colonization or bacterial settling

following surface conditioning and after their

colonization, bacteria start to produce EPS,

(iii) secondary colonization to bacterial layer and EPS

pool by eukaryotic unicellular microorganisms, mainly

protozoan, microalgae and cyanobacteria and (iv) settling

of eukaryotic multicellular organisms as a function of

nutrient sharing, grazing and predation. According to

(Wetzel, 1983), associated organisation from secondary

colonization onwards can be designated as „periphyton‟.

In that way, it could be defined as an advanced

successional stage of biofilm. However, there could be

a fifth (v) phase; the tertiary colonization where

bacterioplankton colonized on the surfaces of unicellular

and filamentous secondary colonizer (e.g. diatom,

Oedogonium etc.). Several bacteria different from early

colonizer settle on algal surfaces at this stage

(Alldredge et al., 1993; Armstrong et al., 2000).

Periphyton are rich Carbon source

TEP with rich Carbon sources of

Glucopolysaccharides in aquatic environments

initiates early colonization of bacteria through

surface conditioning (Stoderegger and Herndl, 1999).

The bacterial EPS from early biofilm exists as a part of

dissolved organic matter (Lignell, 1990) as well as

particulate matter (Decho, 2000). It acts both as rich

organic Carbon storage (Freeman and Lock, 1995) and

Saikia et al., 2013


chief supplier of Carbon demand for organisms that feed

on periphytic aggregates (Decho and Moriarty, 1990;

Hoskins et al., 2003). Being polyanionic in nature

(Costerton et al., 1978), EPS further permits inorganic

nutrient entrapment through ion exchange processes

(Freeman et al., 1995) leading to storage of organic

Carbon in the biofilm. In addition, among the bacterial

fractions, cyanobacteria are important primary producers

and many of their species can fix atmospheric Nitrogen

(Whitton and Potts, 1982). Chemical screening of

laboratory grown, commercially viable cyanobacteria

have revealed that they have a high nutritional value, in

terms of protein (Choi and Markakis, 1981).

During tertiary phase of periphyton development,

algal communities play indirect role in nutrient addition

to periphytic complex through their surfaces. A study on

algae-bacteria interactions on biotic surfaces revealed

that bacterial abundance is significantly higher in areas

of diatom colonization on substrates (Donnelly and

Herbert, 1999). These bacteria contribute to the

management of community metabolism of periphytic

matrix and can trap the metabolic products released

by bacterta on algal surface (Makk et al., 2003). Such

algae-bacteria interactions enrich periphytic organic

matrix with components of polysaccharides, proteins,

nucleic acid and other polymers (Davey and O‟toole,

2000).

Periphytic pathway of nutrient transfer

The periphytic nutrient transfer pathway (PNP)

mainly involves ambient nutrient entrapment, storage

and transferring it to immediate higher trophic level. The

fate of PNP gets its initiation from the surface

conditioning phase of periphyton formation. As soon as

TEP prepares the substrate surface for colonization,

bacteria as initial colonizer develops micro-colonies

(Costerton, 1984) and through EPS, it supplies a

significant source of Carbon to periphytic complex

(Hobbie and Lee, 1980) (Figure 2). A PNP establishes

between dissolved organic in periphytic complex and

inorganic substances in the water column and the higher

trophic levels of the ecosystem (Hynes, 1970). In

general, the Carbon reserve of periphyton generates

through three mechanisms. The first mechanism supplies

energy through bacterial EPS. Bacterial EPS is rich in

carbohydrate, and some time vitamins and other

nutrients. During first-cryptic growth, the dying bacteria

“leak” metabolizable energy to immediate environment

(i.e. EPS) acting as nutrient source to neighbouring

periphyton strata. This property not only protects the

neighbours from starvation but also permits their

multiplication (Postgate, 1976). In a growing periphytic

assembly, cynobacteria and other early algal colonizers

share this Carbon source. In aged periphytic assembly,

the old mostly filamentous periphytic layer receives such

Carbon from overlying bacterial composition resulted

from tertiary phase of colonization. The second

mechanism consists of endogenous energy reserves.

These reserves consist of Carbon that is accumulated and

assimilated inside the microbial cell and can be

Saikia et al., 2013


Figure 1. Formation of periphytic complex on

natural substrate showing tertiary phase of coloni-

zation (Modified from Whale, 1989). TEP, Trans-

parent Exopolymer Prticles; EPS, Extracellular

Polymeric Substances

mobilized to ensure survival during starvation (Dawes

and Senior, 1973) and thereby recovery of periphytic

aggregates due to senescence. The third mechanism of

organic Carbon storage is the polysaccharide exudates

(Freeman and Lock, 1995) released by algae at tertiary

phase under nutrient (especially Phosphorus) limited

condition. The algal components release polysaccharide

exudates to EPS under Phosphorus limitation on which

tertiary phase bacteria flourish. In return, these bacteria

remineralize Phosphorus for algae. In addition, the ECM

with polyanionic by nature (Costerton et al., 1978) is

believed to permit nutrient entrapment through ion

exchange processes (Freeman et al., 1995). (Freeman

and Lock, 1995) proposed that the entrapment

mechanism may also permit the storage of organic

Carbon in the biofilm.

In transferring nutrient through PNP, the

bacterial Carbon enters to organisms in the next trophic

level as complex Carbon rich compounds. The Fatty acid

(FA) component of algae is under extensive research

now a day as Carbon rich compounds. Periphytic matrix

is dominated by algae and hence FA contributes to the

food quality of matured periphytic organization. In algae,

FA increases as a result of exposure to stressful

environmental conditions, such as high temperature,

nutrient extremes and harsh light conditions.

Polyunsaturated fatty acids (PUFAs) also affect many

physiological processes in living organisms and are

major nutrient constituents of polar lipids, and are

present in cell and chloroplast membranes. The

dominance of algae in periphytic canopy acts as rich

source of FA to animals grazing on periphyton.

Primary productivity of periphyton

The energetic relation of an ecosystem is

principally regulated by primary production. In aquatic

ecosystem, algae are dominant primary producers, and

responsible for both Carbon fixation and sequestration.

Periphyton with majority of algae might have significant

contribution to primary production of aquatic ecosystem.

However, very few investigations have been performed

on measurements of photosynthetic rates of algal

periphyton under natural conditions. (Wetzel, 1963)

pointed out technical/methodological difficulty in

assessing such parameters of periphyton under natural

condition. From an analysis on nutrient limiting and

nutrient rich lakes, it is obvious that periphyton

productivity contributes more than 30% of primary

productivity to the aquatic ecosystem (Figure 2a). On

comparison, it seems evident that the nutrient limited

aquatic ecosystems have more or less equal primary

productivity levels to nutrient rich aquatic ecosystems.

The same is not true in case of phytoplankton

(Figure 2b).

Saikia et al., 2013


Figure 2. (a) Primary productivity of Phytoplankton, Periphyton and Macrophytes from

aquatic ecosystems. (b) Primary productivity of Phytoplankton, Periphyton and Macrophyte

in nutrient limited (NL, n=17) and nutrient rich (NR, n=10) aquatic ecosystems. Data from

Vadeboncoeur and Steinman (2002).

An

nu

al

Prim

ary

Pro

du

cti

on

in

percen

tag

e

Phytoplankton Periphyton Macrophyte

Nutrient Regulated Biotic Interactions of Periphyton

Biotic interactions in aquatic ecosystems are

more complex than any other ecosystems for its variable

nature. Interactions between periphyton and biotic

components in aquatic ecosystem are primarily regulated

by nutrients and can be discussed under following

subheadings-

Plankton-periphyton interaction

Periphyton-macrophyte interaction

Grazer-periphyton interaction

Plankton-Periphyton interaction

The plankton-periphyton interaction is

principally regulated by light and nutrient availability in

the environment. Both the communities are composed of

common members of bacterial, algal and zooplanktonic

origin. However, on spatial ground, habitats of both

plankton and periphyton have differences in receiving

light and nutrients. Conceptual models revealed that

nutrient limited environments are dominated by

periphyton than plankton (Wetzel, 2001; Hansson, 1992).

Nutrient limitation results thin planktonic cover that

allows maximum light to pass through water column to

reach the bottom of the ecosystem facilitating

multiplication of periphytic population. Conversely,

plankton rich aquatic ecosystems limit growth of

periphyton due to limited light availability. Epiphytic

communities can better adsorb nutrients from sediments

or bottom of the system through macrophytes

(Burkholder and Wetzel, 1990).

Periphyton-substrate interaction

Substrate type plays a driving role in growth and

succession of periphyton. Being a substrate based

organization, periphyton have access to both organic

nutrients from substrate and inorganic nutrients from

water column. In nutrient rich environments, it receives

nutrients from water column (Eminson and Moss, 1980;

Burkholder, 1996). Here, similar to planktonic cells,

periphytic cells can use inorganic nutrients efficiently,

specifically dissolved organic Phosphorus and in nutrient

limited environments, it relies mainly on organic

nutrients from natural substrate. All artificial substrates

cannot serve as organic nutrient supplier to periphyton.

Substrates like sediments or seed grains acts as nutrient

diffusing substrate releasing nutrients to overlying

periphytic layer. (Hansson, 1989) showed that epipelion

can significantly lower nutrient availability in the water

column due to uptake of diffusing nutrients. (Hagerthey

and Kerfoot, 1998) demonstrated that inflowing ground

water is a significant source of nutrients for episammon

in nutrient limiting environment. These sediments act as

better nutrient source for periphyton (Burkholder, 1996).

Substrate based nutrient uptake by periphyton is further

related to depth, light availability, physical disturbances

etc.

Grazer-Periphyton interaction

Studies reported that several herbivore types

(e.g. gastropods, trichopteran larvae and fish) can

dramatically reduce periphytic biomass to only a few

percent of total biomass (Hillebrand et al., 2000).

Although grazing results reduction in periphytic biomass,

the total productivity of the periphytic complex increases

due to reduced competition among algal members

(Carpenter, 1986; Mc Cormick and Stevenson, 1989).

(Norberg 1999), using transparent incubation chambers,

measured a 4-fold increase in periphyton specific

productivity in grazed periphyton compared to ungrazed

controls. Moreover, the grazer presence increased the

Chlorophyll: biovolume ratio, especially reported from

streams (Hill and Knight, 1987). In addition to increase

in productivity, grazing and competition can modify the

species composition of periphytic algal assemblages

(Duffy and Hay, 2000; Nielsen, 2001), generating

heterogeneity through temporal or spatial scale on the

substrate. A top down effect of consumers on their prey

can be further accelerated by grazer and grazer excretion

of nutrients, removal of senescent cells, or increased

uptake of nutrients by the remaining cells (Lamberti

et al., 1987; Kahlert and Baunsgaard, 1999). Grazers

Saikia et al., 2013


may have strongest effects on Carbon:Phosphorus

and Nitrogen:Phosphorus, but Carbon:Nitrogen and

Carbon:Chlorophyll may remain unaffected (Hillebrand

and Kahlert, 2001). Hillebrand et al., (2008) described

three pathways for grazer mediated periphytic

interactions affecting nutrient stoichiometry. First, the

non algal component, which could be a dominant part

of the organic material of periphyton assemblage

(Frost et al., 2002) is reduced by unselective grazing.

Benthic invertebrates graze upon both detritus and algal

component of periphyton but only algae regenerate.

Therefore, grazing not only reduces non algal component

of periphyton, but also facilitates the growth of live

component within it. (Jones et al., 1999) suggested that

epiphytes can influence the nutritional quality of the

periphyton which grows on their surfaces, making it

more nutritious for grazing by invertebrates, particularly

snails. In return, these grazers might preferentially feed

on the periphyton and clear the plants of a potential

competitor for nutrients, with the plants and grazers both

gaining from this relationship. Secondly, in streams,

nutrient uptake of intact periphyton mats is often slower

than cell specific uptake rates as boundary effects reduce

the uptake ability of the benthic algae (Riber and Wetzel,

1987; Bothwell, 1989; Burkholder et al., 1990). Grazer

presence alters periphyton architecture, increases

periphytic heterogeneity and relative availability of

nutrients (through reducing Carbon: nutrient ratio) per

unit biomass enhancing periphytic nutrient uptake.

Thirdly, the excretion or egestion of nutrients or both by

grazers also increase the supply of nutrients to the

periphytic assemblages. Grazers may spatially recycle

nutrients that increase the availability and uptake of

nutrients by the periphyton. However, in streams,

grazers may increase the export of nutrients

(Mulholland et al., 1991).

CONCLUSION

Disrupted nutrient cycling is a major problem

both in freshwater and marine ecosystems and

periphyton could be a non-point manager of nutrient

cycle disruption and hence can overplay on plankton for

nutrient cycling in aquatic ecosystem. During renovative

practices, strategies of aquatic ecosystem health

managers greatly ignore the role of these substrate based

microorganisms. At the same time, it can play as an

efficient supplier of nutrient to its grazer under

controlled and well managed productive practices. It is

observed that at traditional level, farmers from different

parts of the world have been practicing periphyton to

feed aquacrops to convert periphytic energy biomass to

crop biomass (Saikia and Das, 2009). Such conversion of

biomass is an outcome of increased assimilation of

micro- and macro nutrients from periphytic complex in

the fish body through trophic upgrading (Saikia and

Nandi, 2010). Further researches on the mode of energy

transfer through periphytic food chain, enhanced nutrient

uptake under manipulative nutrient input, modelling on

applied periphytic ecology, ecotoxicology, Carbon

entrapment and delivery, directing nutrient and Carbon

sequestration both in marine and freshwater are needed

for better understanding of its role in aquatic ecosystem.

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Jou

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Biology

Assessing heavy metal contamination of road side soil in urban area

Keywords: Pollution, combustion, heavy metal enrichment, road side soils, enrichment factor, Normalized scatter coefficient value, environmental pollution.

ABSTRACT: Environmental pollution of heavy metals from automobiles has attained much attention in the recent past. The pollution of soil by heavy metals is a serious environmental issue. Heavy metals are released during different operations of the road transport such as combustion, component wear, fluid leakage and corrosion of metals lead, cadmium, copper and zinc which are the major metal pollutants of the road side environment. The present research is conducted to study heavy metal contamination in road side and industrial soil of Madurai city. The soil samples are collected from three sites and analyzed for six heavy metals (Pb, Cu, Cr, Zn, Ni and Cd). Their concentration and distribution in different depths (0 cm, 5 cm and 10 cm) were determined. Heavy metal contents were analyzed by Atomic Absorption Spectroscopy (AAS). The studies with Enrichment Factor (EF) indicate that lead has been enriched to quite great extent while the Normalized Scatter Coefficient values (NSC) indicate faster enrichment of cadmium. The level of heavy metals in road side soils were higher as compared to their natural background levels. The results revealed that the heavy metals are harmful to the road side vegetation, wild life and the neighbouring human settlements.

789-796 | JRB | 2013 | Vol 3 | No 1

This article is governed by the Creative Commons Attribution License (http://creativecommons.org/

licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and reproduction in all medium, provided the original work is properly cited.


An International Scientific

Research Journal

Authors:

Sarala Thambavani D1,

Vidya Vathana M2.

Institution:

1. Associate Professor

Department of Chemistry,

Sri Meenakshi Govt. Arts

College (W), Madurai.

2. Assistant Professor,

Department of Chemistry,

Sacs M.A.V.M.M Engg.

College, Madurai.


Sarala Thambavani D.


Dates: Received: 16 Jan 2012 Accepted: 27 Jan 2012 Published: 16 Feb 2013

Article Citation: Sarala Thambavani D and Vidya Vathana M. Assessing heavy metal contamination of road side soil in urban area. Journal of Research in Biology (2013) 3(1): 789-796

Journal of Research in Biology An International Scientific Research Journal

Original Research

INTRODUCTION

Pollution in recent years has increased

considerably as a result of increasing human activities

such as burning of fossil fuels, industrial and automobile

exhaust emissions. The pollution of soils by

heavy metals from automobile sources is a serious

environmental issue. The majority of the heavy metals

are toxic to the living organisms and even those

considered as essential can be toxic if present in excess.

The heavy metals can impair important biochemical

processes posing a threat to human health, plant growth

and animal life (Jarup 2003; Michalke 2003; Silva et al.,

2005).

The waste products from vehicles that ply

highways contain some heavy metals inform of smokes.

Emissions from exhaust pipes of automobile engine and

contacts between different metallic objects in machines

contain such heavy metals as Lead (Pb), Zinc(Zn), Iron

(Fe), Copper (Cu), Chromium (Cr) and Cadmium (Cd)

and are major sources of pollution among soils (Turer

and Maynard, 2003).

Soils are usually regarded as an ultimate sink.

For heavy metals discharged into the environment (Banat

et al., 2005) and sediments can be sensitive indicators for

monitoring contaminants in aquatic environment (Pekey

et al., 2004). Therefore the environmental problem of

soil and sediment pollution by heavy metals has received

increasing attention in the last few decades in both

developing and developed countries throughout the

world (Zhang et al., 2007). Hence, inorder to monitor

heavy metal pollution in an area, due to the

anthropogenic activity (Sarala Thambavani and Vathana,

2011), the soil samples represent an excellent media

because heavy metals are usually deposited in the top

soil (Govil et al., 2001; Romic and Romic, 2003) and

help in knowing the sources of heavy metals and also

controlling and optimizing their effects on the human

health.

It is needless to say that the industrial activities

in the metropolitan cities of the world are responsible for

the addition of pollutants through chemical factories,

residential activities (Point sources) and vehicular traffic

(non-point sources) which are the primary sources of soil

pollution. The objective of this study, is to investigate the

effect of heavy metal pollution of soil along road sides.

The present study reports the role of industrial and

urban activities in the heavy metal contamination of the

soils in the Madurai industrial area with the objectives:

To assess the extent of heavy metal pollution

influenced by urban and industrial activities.

To predict the rate of heavy metals pollution in the

future if the activities are allowed with the same

pace.

To understand the variations in the behavior of

different heavy metal.

METHODS

Field Methodolgy

To understand the state of environment of the

Madurai area a detailed field survey was carried out and

after having identified possible sources of pollution a

part of Madurai area was selected. This area is under

intense human interference in terms of growing

urbanization (municipal sewage sludge, traffic pollution

in particular) and industrialization.

Selection of sampling site

In the present study stratified regular sampling

method was adopted for soil sample collection as in

geo-assessment of the variables estimated, the stratified

regular sampling is more suitable because this kind of

sampling draws homogenous error (Burgess et al., 1981).

Different sampling stations were selected and samples

are collected from the top layer of the soil using plastic

spatula after removing the debris, rock pieces and

physical contaminants. In order to have the background

concentration values of the heavy metal elements, three

soil samples were collected, each from 100 cm below

Thambavani and Vathana, 2013


ground level, which are least affected by anthropogenic

activities (Table1). The samples were placed in the clean

polythene bags, which were brought to the laboratory.

Laboratory Methodology

The samples were brought to the laboratory

where they are dried and mixed thoroughly to obtain the

representative samples. Soon after drying the debris and

other objects were hand picked up and the sample were

grounds in a mortar to break up the aggregates or lumps,

taking care not to break actual soil particles. Soil samples

were then passed through a 2 mm sieve in order to

collect granulometric fraction. Since trace metals are

often found mainly in clay and silt fractions of soil and

hence the size fraction <63 µm sieve (wet sieving) and

was used to measure the concentration of the heavy

metals Lead, Copper, Chromium, Zinc, Nickel and

Cadmium from all the samples collected.

For this purpose the clay and silt fraction were

digested by acids to get the solution by taking 5 g of

sample into a 300 ml polypropylene wide-mouthed jar

and distilled water was added to make a total 200 ml.

Then it was acidified with 10 ml HF, 5 ml HClO4,

2.5 ml HCl and 2.5 ml HNO3 in order to completely

digest the soil. This jar was shaken on an orbital shaker

for 16 h at 200-220 rpm before being filtered through

whatman filter paper (No.42) into acid washed bottles.

The solution was stored and heavy metal contents were

analyzed by Atomic Absorption Spectrophotometer as

per the method recommended by committee of soil

standard methods for analyses and measurement (1986).

The raw data obtained during the course of laboratory

analyses were stored in Microsoft Excel software and

further processed to obtain various parameters required

for interpretation.

RESULTS

The concentration of heavy metals Lead,

Copper, Chromium, Nickel and Cadmium in the soils

of Madurai industrial, traffic and residential area

were analyzed, collected at six sampling stations during

May 2011- Oct 2011. The range of the concentrations

found in different sampling stations are (i) Pb industrial

(24.81-42.37 mg/kg), traffic (26.80-5.32 mg/kg)

and residential (20.42-2.66 mg/kg) (ii) Cu industrial


and residential (10.5 -18.16 mg/kg) (iii) Cr industrial


and residential (25.12 mg/kg) (13.60-18.52 mg/kg)

( i v) Zn in dus t r i a l (22 . 5 -45 . 6 mg/kg) ,

traffic (22.32-25.46 mg/kg) and residential (22.24- 25.12

mg/kg) (v) Ni industrial (11.85-14.0 mg/kg), traffic

( 11.52 -14.80 mg/kg) and residential (11.70-13.9 mg/kg)

(vi) Cd industrial (1.24-4.32 mg/kg), traffic

(1.60-3.62 mg/kg) and residential (1.70-2.25 mg/kg).

The mean concentration for these heavy metals

from the surface soil have been calculated to be (i) Pb

industrial (33.23), traffic (41.50) and residential (24.31).

(ii) Cu industrial (12.97), traffic (15.03) and residential

(14.98). (iii) Cr industrial (24.33), traffic (17.53) and

residential (15.51). (iv) Zn industrial (29.78), traffic

(24.23) and residential (23.74). (v) Ni industrial (12.77),

traffic (13.72) and residential (12.99). (vi) Cd industrial

(2.94), traffic (2.59) and residential (1.92) respectively at

the confidence limits of 95%.

The concentration of heavy metals in all the

sampling stations exhibit an increasing trend over a very

short period of monitoring from May 2011-Oct 2011

(Figure 1). It was observed that the mean concentration

of Lead has been increased in all the three sampling



Table 1 Natural Local background concentration values (mg/kg) of the heavy elements of soils

Sampling stations Pb Cu Cr Zn Ni Cd

Industrial Area 5.14 9.44 9.89 11.32 11.28 0.32

Traffic Area 5.22 9.58 10.09 11.76 11.29 0.30

Residential Area 5.26 9.63 11.10 11.87 11.31 0.35

stations followed by Zinc, Chromium, Copper, Nickel

and Cadmium.

Accumulative Signature of Heavy Metals

An increasing trend has been found for the heavy

metal elements Lead, Copper, Chromium, Zinc Nickel

and Cadmium wherein the Lead and Cadmium are

getting accumulated with very rapid rate mainly due to

anthropogenic activities (Sayadi, 2009). In order to

assess the variations in the heavy metal accumulations in

the soils, the calculated measures that is Enrichment

Factor and Normalized Scatter Coefficient were used.

The Enrichment Factor (EF) is a ratio of the

concentrations of the heavy metals in the soil samples to

the corresponding concentration of natural background

concentration. EF is calculated with the help of the

formula given by Subramanian and Datta dilip (1998)

and presented in Table 2.

Normalized Scatter Coefficient (NSC) has been

calculated to asses the temporal variability of the heavy

metals in the soils. It helps to understand the increasing

or decreasing concentration of heavy metals in the soils

with the passage of time which is independent of the past

focusing only at the period of study. The NSC for any

element is calculated (Table 3) with the following

formula (Sayadi and Sayyed, 2010).

The NSC values + 100% indicates absolute

increase while-100% means absolute decrease. The value

of 0% can be regarded for no change in the parameters

under consideration.

DISCUSSION

In order to evaluate the rate of accumulation of

heavy metals in the soils the mean values for all heavy

metals studied were considered along with Enrichment

factor values of all six metals (Table 2), which clearly

indicate the highest enrichment of Cadmium followed by

Lead, Zinc, Chromium, Copper and Nickel in all the

three sampling stations of industrial, traffic and

residential area. The values of NSC for all six heavy

metal showed that Cadmium is increasing in soil

environment of industrial area followed by Zinc,

Chromium, Lead, Copper and Nickel. In traffic area

Lead is increasing in soil environment followed by

Cadmium, Chromium, Copper, Nickel and Zinc and in

residential area Copper is increasing in soil environment

followed by Chromium, Cadmium, Lead, Nickel and

Zinc.

It is observed that in all the sampling sites, Lead

shows highest concentration in soil and also have high



Pb Cu Cr Zn Ni Cd

4.6 1.1 1.7 1.9 1.1 3.8

5.3 1.1 1.8 2.1 1.1 4.1

5.7 1.4 2.2 2.3 1.0 8.8

6.9 1.5 2.7 2.5 1.1 11.8

7.7 1.5 2.9 2.9 1.2 13.1

8.3 1.7 3.5 4.0 1.2 13.5

Industrial Area

Pb Cu Cr Zn Ni Cd

5.1 1.1 1.4 1.9 1.0 5.3

5.6 1.4 1.6 1.9 1.2 6.6

6.9 1.5 1.6 2.1 1.2 8.3

8.7 1.7 1.8 2.1 1.3 9.5

10.1 1.8 1.8 2.1 1.3 9.9

11.2 1.9 2.1 2.2 1.3 12.1

Traffic Area

Pb Cu Cr Zn Ni Cd

3.9 1.1 1.2 1.9 1.0 4.9

4.2 1.3 1.3 1.9 1.1 4.9

4.7 1.3 1.3 2.0 1.2 5.3

4.8 1.8 1.4 2.0 1.2 5.7

5.0 1.9 1.5 2.1 1.2 5.7

5.1 1.9 1.7 2.1 1.2 6.4

Residential Area

Table 2 Enrichment Factor for heavy metals in the

soils

concentration in the last sampling –

concentration in first sampling

NSC = x 100

concentration in the last sampling +

concentration in first sampling

EF = Value of a given metal concentration found on soil (mg/kg)

Natural local background concentration of the metal (mg/kg)

enrichment factor. Cadmium shows lowest concentration

in soil but is has quite high enrichment factor, while

Copper, Chromium, Zinc and Nickel shows higher metal

concentration but rather low EF when compared to lead.

The scatter plot of the mean concentration of

heavy metals was plotted against the EF for all the three

sampling sites (Figures 5,6,7). Per usual of the result

showed that Zinc is having high mean concentration but

it is not getting enriched in proportion to its mean

concentration. On the other hand Cadmium though

having lowest mean concentration has higher rate of

enrichment. Lead shows the highest mean concentration

and also corresponding highest enrichment factor.

The behavior of Zinc may be attributed to its

source mainly from weathering of the parent rock while

that of Cadmium and Lead mainly due to anthropogenic

activities. EF normally reveals the addition and or

removal of metal under consideration which is a result of

cumulative activity in the region. Hence the Enrichment

factor should denote the total enrichment and or

depletion of an element and cannot evaluate the trend for

the short term accumulation.

When the mean values of EF and NSC for all the

six heavy metals are studied at all the sampling stations

(Figures 8, 9,10) it can be stated that Cadmium has been

enriched to a quite greater extent followed by Lead, Zinc,

Chromium, Copper and Nickel at all the sampling sites.

On the other hand the Normalized Scatter Coefficient

value indicates that Cadmium has got enriched in faster

rate at industrial area followed by Zinc, Chromium,

Lead, Copper and Nickel. In traffic area Lead is getting

enriched in the faster rate followed by Cadmium,

Chromium, Copper, Zinc and Nickel. But in residential

area, the NSC value indicate that Cu is quite enriched

with the faster rate followed by Chromium, Cadmium,

Lead and Zinc.



Figure 2. Industrial Area

Figure 3. Traffic Area Figure 4. Residential Area

Figure 1. Mean Concentrations of the heavy metals

on different sampling stations

CONCLUSION

The variation assessment of heavy metal

pollution by using Enrichment Factor and Normalized

Scatter Coefficient in the soil sample collected from the

study area between May 2011-Oct 2011 has revealed

significant increase in the six heavy metals (viz Pb, Cu,

Cr, Zn, Ni and Cd). Enrichment Factor values shows

that Cadmium has enriched to a greater extent followed

by Lead, Zinc, Chromium, Copper and Nickel.

Normalized Scatter Coefficient value indicate that Lead

is getting accumulated in a faster rate followed by

Cadmium, Chromium, Copper, Zinc and Nickel. In

summary the soils in the Madurai industrial, traffic and

residential area are significantly contaminated by heavy

metals and hence more attention to be paid to heavy

metal pollution particularly for Lead and Cadmium. In



Figure 7. Residential Area

No

rm

ali

zed

Scatt

er C

oeff

icie

nt

%

En

ric

hm

en

t F

acto

r

Figure 8 INDUSTRIAL AREA

No

rm

ali

zed

Scatt

er C

oeff

icie

nt

%

Figure 9 TRAFFIC AREA

En

ric

hm

en

t F

acto

r E

nric

hm

en

t F

acto

r

No

rm

ali

zed

Scatt

er C

oeff

icie

nt

%

Figure 10. RESIDENTIAL AREA

Figure 5. Industrial Area

Figure 6. Traffic Area

order to prevent heavy metal contamination in the soils

from the Madurai city and to maintain the ecological

balance some immediate measures as per environmental

quality criteria, a need to be taken.

REFERENCES

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Pb Cu Cr Zn Ni Cd

26.1 21.9 33.8 33.8 8.6 55.4

21.1 21.1 30.9 31.7 8.1 53.2

17.8 12.0 23.6 26.7 6.6 21.3

8.9 7.8 13.0 24.3 4.7 6.7

3.4 7.6 9.8 16.8 1.9 1.4

0 0 0 0 0 0

Industrial Area

Pb Cu Cr Zn Ni Cd

13.2 26.4 15.3 6.1 8.9 13.9

8.8 17.9 13.1 4.7 7.2 13.4

3.9 16.8 11.9 3.4 3.2 9.8

2.5 1.6 8.7 2.6 2.5 6.1

0.8 0.6 5.4 0.4 0.6 5.6

0 0 0 0 0 0

Residential Area

Pb Cu Cr Zn Ni Cd

37.0 25.9 19.5 6.6 12.5 38.7

32.9 14.6 15.8 4.9 5.7 29.1

23.0 10.9 14.0 1.9 2.7 18.3

12.5 5.5 8.0 0.9 2.0 12.1

5.2 4.1 7.3 7.3 0.8 9.5

0 0 0 0 0 0

Traffic Area

Table 3 Normalized Scatter Coefficient (%)of the

heavy metals in the soils of the study area

of the Gangae - Brahmaputra-Meghna river system in the

Bengal basin.Environmental Geology 36(1-2):93-101.

Turer D and Maynard JB. 2003. Heavy metal

contamination in highway soils. Comparison of Corpus

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key to mobility. Clean Technology and Environmental

Policy 4:235-245.

Zhang LP, Ye X, Feng H. 2007. Heavy metal

contamination in Western Xiamen Bay Sediments

and its Vicinity, China. Marine Pollution Bulletin

54(7):974-982.




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