Journal of Research in Biology - Volume 3 Issue 1

Journal of Research in Biology An International Open Access Research Journal
Original Research
Anatomical variation in the olfactory apparatus of marine teleosts

Journal of Research in Biology
Authors: ABSTRACT:
Biswas S1, Datta NC2,
Sarkar SK1 and De SK1.
The olfactory apparatus of marine teleosts viz., Rastrelliger kanagurta,
Institution: Scomberoides commersonianus and Platycephalus scaber belonging to the family
1. Department of Zoology,
of Scombridae, Carangidae and Platycephalidae respectively has been fixed in
Vidyasagar University,
10% formaldehyde solution for 24 h and anatomically examined under light
Midnapore (West) - 721102,
microscope (LM). Anatomical variation regarding the nostrils, olfactory rosette,
West Bengal, India.
occurrence of accessory nasal sacs, olfactory lobes, length of the olfactory nerve
2. 110/20 B. T. Road, tracts, etc. are observed. These morphological variations may denote species specific
Kolkata - 700108, West and may decisive for several biological functions.
Bengal, India.
Corresponding author:
Subrata Kumar De.
Email: Keywords:
skdvu@yahoo.co.in Olfactory, Rosette, Scombridae, Carangidae, Platycephalidae, etc.
Phone No: Article Citation:

+91 03222 275329 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
Dates:
Web Address: Received: 08 Nov 2012 Accepted: 20 Nov 2012 Published: 09 Jan 2013
http://www.jresearchbiology.com/
documents/RA0304pdf.
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.
742-748 | JRB | 2013 | Vol 3 | No 1

An International Open Access
Research Journal www.jresearchbiology.com
Biswas et al., 2013
INTRODUCTION and Platycephalus scaber (Linnaeus, 1758), an estuarine

Olfaction is an important type of chemoreception amphidromous fish to unfold their structural components
which is mediated through well developed, complex and and functional significance in olfaction.
organized olfactory system in vertebrates (Freitag et al.,
1999). This system generally develops from the MATERIALS AND METHODS
specialized tissue, called olfactory placode (von Kupffer, Adult, sex-independent specimens of
1894). In the early stage of development, placodes are R. kanagurta, S. commersonianus and P. scaber were
formed from the preplacodal ectoderm of the anterior collected from the coastal regions of Bengal, India.
region of the embryo, medial to epidermis and lateral to Specimens were preserved in 10% formaldehyde
both neural crest and neural plate (Knouff, 1935). solution for 24 h and brought to the laboratory. The
The developmental process leading to the formation of olfactory apparatus of these species were dissected out
fish olfactory organ, is little diverse (Hansen and and separately mount on grease free slide by using
Zielinski, 2005). The peripheral olfactory organ is the glycerine. The olfactory apparatus of respective species
first chemosensory organ to develop in fish preceding the was examined under light microscope (LM).
systems of solitary chemosensory cells (Kotrschal et al.,
1997) and taste (Hansen et al., 2003). Fish generally RESULTS
possesses a paired olfactory organ located at the anterior R. kanagurta (Figure 1A) possesses two pairs of
part of the head (Derivot, 1984; Hansen and Zeiske, nostril viz., anterior nostril and posterior nostril
1998; Dving, 2003; De and Sarkar, 2009). The olfactory (Figure 1B). The anterior nostril is an oval shaped
chambers, olfactory rosette, accessory nasal sacs, structure, encircled by a thick lip like ridge of skin and
olfactory bulbs, olfactory nerve tracts and brain are the located at the dorsal region of the snout where as the
major components of fish olfactory apparatus posterior nostril is situated in front of the eye.
(Hamdani and Dving, 2007). The anatomy of the R. kanagurta shows a slit like structure i.e., posterior
olfactory apparatus shows wide range of structural nostril which is located at a moderate distance from the
diversity regarding the shape and size of the olfactory anterior one (Figure 1B). The olfactory apparatus of the
rosette, number of the olfactory lamellae, occurrence of said species is present at the antero-dorsal side of the
accessory nasal sacs, etc. among the teleostean groups snout in between the anterior and posterior nostril
(Kleerekoper, 1969). The anatomical details of the (Figure 1C). The multilamellar olfactory rosette is oval
olfactory apparatus in several species belonging to the in shape and is situated at the floor of the olfactory
diverse teleostean taxa with respect to their habitat are chamber (Figures 1D and 1E). The triangular olfactory
still an obscure part in sensory biology. The sensory lamellae vary from 60 to 70 in number per rosette and
systems of fishes show notable adaptations according to are arranged pinnately on the raphe (Figure 1E). The
habitat and mode of life in comparison with the higher accessory nasal sacs are clearly marked in the olfactory
vertebrates (Bone and Moore, 2008). The present study apparatus of R. kanagurta. The lacrimal sac is conical in
focused on the true anatomy of the olfactory apparatus in shape and located at the antero-medial region of
three species of different ecological habitat viz., lachrymal bone in association with olfactory rosette
Rastrelliger kanagurta (Cuvier, 1816), a marine (Figures 1C and 1D). The ethmoidal sac is connected at
oceanodromous fish; Scomberoides commersonianus the posterior part of the olfactory rosette and situated
(Lacepede, 1801), a brackish water amphidromous fish within a groove at the antero-lateral extension of the
743 Journal of Research in Biology (2013) 3(1): 742-748
Biswas et al., 2013
Plate - 1
OLR
LS
ETS
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).
frontal bone (Figures 1C and 1D). The olfactory nerve (Figure 2B). The olfactory rosette is circular in shape and
tracts are arised from the base of olfactory rosette and the consisting of two pairs of olfactory lamellae
length varies from 16 mm to 18 mm respectively. The (Figures 2C and 2D). The number of the olfactory
distal part of the olfactory nerve tracts are connected lamellae ranges from 100 to 140 per rosette (Figure 2D).
with olfactory lobes of the brain. The size of the The accessory nasal sacs are not distinct in the olfactory
olfactory lobes is comparatively small in R. kanagurta apparatus. The olfactory nerve tracts are arised from the
(Figure 1C). base of the olfactory rosette and the length is ranges from
In S. commersonianus (Figure 2A), the nostrils 10 mm to 12 mm. The olfactory lobe is comparatively
are closely associated. The anterior nostril is an oval large in size (Figure 2C).
shaped aperture and partly guarded by a nasal flap where Interestingly, the nostrils of P. scaber
as the posterior nostril is crescentric in shape (Figure 3A) are situated at the antero-dorsal region to the
Journal of Research in Biology (2013) 3(1): 742-748 744

Biswas et al., 2013
Plate - 2
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).
eye and lying far apart from each other. The anterior DISCUSSION
nostril is oval in shape and has a tongue like nasal flap Olfactory systems of fish are among the most
where as the posterior nostril is valvular (Figure 3B). highly developed olfactory senses of vertebrates
The olfactory rosette is relatively large in size and oval (Kleerekoper, 1969). The sense of olfaction is mediated
in shape (Figures 3C and 3D). The number of the through olfactory apparatus associated with nostrils. The
olfactory lamellae varies from 50 to 76 in number per nostrils are responsible for incurrent and excurrent of
rosette (Figure 3D). The absence of accessory nasal sacs water during water ventilation (Nevitt, 1991). The
is noted in the olfactory apparatus of P. scaber. The structure of the anterior and posterior nostril varies
length of the olfactory nerve tracts ranges from among the teleostean fishes (Kapoor and Ojha, 1972).
22 mm - 24 mm. and it is well connected with the The presence of nasal flaps in between the both nostrils
olfactory lobe of the brain (Figure 3C). 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
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).
olfactory rosette perhaps adopt several type of to sniff (Nevitt, 1991; Cox, 2008). R. kanagurta
arrangement pattern (Holl, 1965). This lamellar possesses well developed accessory nasal sacs but
arrangement may help in the particular sensitivity to S. commersonianus and P. scaber has no accessory nasal
certain components like amino acids, steroids, sacs. The movement of jaws and its associated muscles
prostaglandins, etc. (Theisen et al., 1991). Anatomically are also very significant for the water ventilation
the lamellar surface in S. commersonianus is much closer (Nevitt, 1991). Accessory nasal sacs may be found in the
due to the short distance between anterior and posterior olfactory organs of fishes with widely variable life styles
nostril than R. kanagurta and P. scaber. Therefore, the and habitats, both marine and fresh water which are not
water soluble odorants may travel short distance to confined to one particular situation (Cox, 2008).
interact with comparatively greater olfactory lamellar The olfactory nerve may convey the chemical cues
surface of S. commersonianus. The water ventilation is during water ventilation to the brain (Hamdani and
assisted by the pumping mechanism of accessory nasal Dving, 2007). The olfactory information plays an
sacs (Theisen et al., 1991) and may provide the ability important role in different behaviour of fish such as

Biswas et al., 2013
searching of foods, avoidance of predators, Cox JPL. 2008. Hydrodynamic aspects of fish olfaction.
discrimination between individuals of the same and Journal of the Royal Society Interface, 5(23):575-593.
different, parental care, orientation in migration, etc.
Derivot JH. 1984. Functional anatomy of the peripheral
(Hara, 1971). The olfactory system of teleosts is highly a
olfactory system of the African lungfish Protopterus
specialized structure for the recognition of various water
annectens Owen: macroscopic, microscopic, and
soluble chemical cues, so this may serve as a biological
morphometric analysis. American Journal of Anatomy,
model to monitor environmental health as well
169(2):177-192.
as specific meagerness of the pollutants. The
xenotoxification of ocean especially the acidification De SK and Sarkar SK. 2009. Morphoanatomy of
may also impair the ability of olfactory discrimination of olfactory apparatus of Pseudapocryptes lanceolatus
coastal and marine species (Munday et al., 2009). Thus, (Bloch and Schneider) Journal Environment and
it is necessary to examine the effect of specific toxic Ecology, 27(4):1646-1648.
agent at a subcellular level of olfactory structures in
Dving KB. 2003. The fish olfactory system: Its role in
marine teleolsts.
normal biology and in toxicological research.
Proceedings of the Seventh International Symposium,
CONCLUSION
Tallinn, Estonia. 149-158.
This comparative anatomical study on the
olfactory apparatus along with brain in three different Freitag J, Beck A, Ludwig, G, von Buchholtz L, Breer
marine teleost belonging to the diverse ecological habitat H. 1999. On the origin of the olfactory receptor family:
shows much structural variation according to the receptor genes of the jawless fish (Lampetra fluviatilis).
changing environment which may be significant for Gene, 226(2):165-174.
ecomorphology and evolutionary aspects of
Hamdani EH, Dving KB. 2007. The functional
neurobiology (Kotrschal et al., 1998). However, the
organization of the fish olfactory system.
olfactory system of marine, estuarine and coastal or
Prog Neurobiol, 82(2):80-86.
migratory species may experience rapid fluctuations of
environmental inorganic ions (Hubbard et al., 2000), so Hansen A and Zeiske E. 1998. The peripheral olfactory
it could be an interesting part to identify the cellular organ of the zebrafish, Danio rerio: an ultrastructural
components that are involved in the ion regulation of the study. Chem Senses, 23:39-48.
olfactory apparatus in these migratory teleosts.
Hansen A, Rolen SH, Anderson KT, Morita Y,
Caprio J, Finger, TE. 2003. Correlation between
ACKNOWLEDGEMENTS
olfactory receptor cell type and function in the channel
Authors are thankful to the Head, Department of
catfish. J Neurosci., 23:9328-9339.
Zoology, Vidyasagar University, West Bengal, for
providing the necessary laboratory facilities. Hansen A and Zielinski, BS. 2005. Diversity in the
olfactory epithelium of bony fishes: development,
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Original Research
The use of purple yam (Dioscorea trifida) as a health-promoting

ingredient in bread making
Authors: ABSTRACT:
Teixeira AP1,
Oliveira IMA1, Lima ES1 The use of purple yam (Dioscorea trifida) was evaluated as possible
and Matsuura T2. 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
Institution:
acceptance test and microbiological analysis of the formulations 10%, 15% and 20%
1. Faculdade de Cincias
were also performed. Except for lipids, the centesimal composition and caloric values
Farmacuticas (FCF),
Universidade Federal do revealed no statistically significant differences. An addition of purple yam in natura up
Amazonas (UFAM), Rua to 20%, instead of wheat flour in ordinary bread (0%), can be made with no effect on
Alexandre Amorim, 330, the diets energy. The free radical scavenging, 2.2-diphenyl-1-picryl-hydrazyl (DPPH)
Aparecida, CEP: 69010-330, and lipid per oxidation (LPO) methods revealed that the greater the percentage of
Manaus, AM, Brasil. 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
2. Instituto de Cincias revealed a significant difference only in the attribute colour. Purple yam breads
Biolgicas (ICB), showed no preferable differences. Results highlight the feasibility of purple yam bread
Universidade Federal do as a health-promoting food in the Amazon region.
Amazonas (UFAM), Av.
General Rodrigo Octvio
Jordo Ramos, 3000, Keywords:
Campus Universitrio,
Purple yam (Dioscorea trifida); antioxidant activity; health-promoting food;
Coroado I CEP: 69077-000,
Amazon region.
Manaus, AM, Brasil.
Corresponding author: Article Citation:

Antonia Paiva Teixeira. Teixeira AP, Oliveira IMA, Lima ES and Matsuura T.
The use of purple yam (Dioscorea trifida) as a health-promoting ingredient in bread
Email: making.
nietapt@yahoo.com.br Journal of Research in Biology (2013) 3(1): 747-758.
Web Address: Dates:
http://www.jresearchbiology.com Received: 15 Nov 2012 Accepted: 27 Nov 2012 Published: 09 Jan 2013
document/ RA0306.pdf.
cited.
747-758 | JRB | 2013 | Vol 3 | No 1

Teixeira et al., 2013
INTRODUCTION pies and porridges. Nevertheless, this species has

Yams belong to the family Dioscoreaceae, genus undergone little scientific investigation, so little is known
Dioscorea (Pedralli, 1988; 1997; Pedralli et al., 2002; about its management techniques, genetic improvement,
Pedralli, 2004). This family is made up by 6 to 9 genera nutritional potential, industrial use, storage procedures,
comprising over 600 species distributed throughout the characterization, uses as natural dye, as well as its use as
Worlds tropical, subtropical and temperate regions a health-promoting ingredient, among others.
(Barroso et al., 1974; Pedralli, 1988; 1997; Melo By and large, the bread consumed throughout the
Filho et al., 2000; Pedralli et al., 2002; Pedralli, 2004). world is made mostly of wheat flour, salt and yeast.
The yams (Dioscorea spp.) yield tubers, which are very Many other ingredients, have been incorporated into
important as staple, nutritional and healthy food, and are bread formulation, so as to increase its diversity and
still used as an ingredient in traditional Chinese herbal product appeals (Hsu et al., 2004).
medicine. They show a worldwide distribution, and are A few studies have highlighted the great
found in many tropical countries, in South-Eastern Asia potential of purple yam in bread making. In this case,
and Western Africa, where the species were introduced yam flour may replace part of the wheat flour, improving
by cultivators (Rasper and Coursey, 1967; Akanbi et al., bread quality, as well as adding economical advantages
1996; Omonigho and Ikenebomeh, 2000; Lin et al., to it (Abramo, 1990; Hurtado et al., 1997; Litvin et al.,
2005). They can also be found in some American 1998; Omonigho and Ikenebomeh, 2000; Ratti, 2001).
countries, particularly in Brazil, where one can find them Hsu et al., (2004) demonstrated the presence
in all regions, from the Amazon down to the Southern of antioxidants in the flour of purple yam
part of the country (Chu and Figueiredo-Ribeiro, 1991; (Dioscorea purpurea), in five formulations of breads
Pedralli, 1997; 2004). prepared with this tubers flour, with excellent
Purple yam (Dioscorea trifida) is an American acceptance in Taiwan supermarkets. Contado et al.,
native species, which was domesticated by Amerindians, (2009) showed yam (Dioscorea spp.) mucilage-based
with the cultivar distribution possibly pointing out its loaf to present good public acceptance as to flavor,
domestication in Brazilian and Guyana border areas, aroma and texture with sensory attributes, demonstrating
followed by dissemination throughout the Caribbean the use of this tuber to be feasible as improvers in bread
islands (Pedralli, 1988; Pedralli et al., 2002; Pedralli, making.
2004). D. trifida shows a wide distribution in Central and The following aspects motivated the use of
South America, from the Caribbean to Peru. In Brazil it purple yam (Dioscorea trifida) in natura as a bread
is found all the way from the Amazon right down to the manufacturing health-promoting ingredient, in the
Southern region. The species is associated to forest present work: 1) its significant world consumption,
environments-Amazonian highland tropical rainforests, presenting a considerable, expanding tillage alternative
Coastal Atlantic Forest in Southeastern Brazil and, (Rasper and Coursey, 1967; Abramo, 1990; IITA, 2007);
mesophytic (seasonable) and gallery forests (Pedralli, 2) although, as yet incipient, an increase on the
1997). production of this tuber in the State of Amazonas, Brazil,
Here in the Amazonian region, purple yam especially in Caapiranga and Careiro Castanho
(D. trifida) may be consumed in the following ways: municipalities is being observed. According to the
baked, boiled, mashed, as ingredients for soups and meat Instituto de Desenvolvimento Agropecurio do Estado
stews, and in the formulation of flour for making cakes, do Amazonas (IDAM) in 2008, 110 families of the
Caapiranga municipality yielded 2,475 tonnes in an area (edible portion), in Amazonas State Townships. The
of 165 ha; and 3) the presence of antioxidants in purple types most easily identified are: roxinho (light purple
yam, which increases the nutritional capacity in breads flesh); roxo (mid purple flesh); roxo (dark purple flesh);
made from this tuber (Hsu et al., 2004). branco (white flesh); and misto (white-purple flesh)
The main aim of the present study was to (Figure 1).
evaluate the potential of purple yam yield in the State of Purple yam samples were collected at two
Amazonas, Brazil as a health-promoting ingredient in Amazonas Townships: Caapiranga and Careiro
bread making. On this context, it determined the Castanho. Due to the seasonality and availability of these
centesimal composition, caloric value, and antioxidant tubers in the region, the centesimal composition analyses
properties of purple yam as well as of breads made from of yams and breads were performed with Caapiranga
this tuber in natura. Then, it undertook an organoleptic samples. Yam and bread antioxidant and bread sensory
characteristic assessment of the breads, following tasters and microbiological analyses were carried out with
panel acceptance criteria. This purple yam species is, for Careiro Castanho samples.
the very first time, being used in the Amazonian region, Purple yam bread elaboration
as a feasible alternative for bread making. On account of the probability of getting breads
with higher antioxidant concentration (Hsu et al., 2004),
MATERIALS AND METHODS roxo (dark purple flesh) type samples were used in the
Species identification and purple yam tuber present study (Figure 1C). Yams in natura, for replacing
(Dioscorea trifida) collection wheat flour, were washed, peeled, weighed, ground
Identification of the species Dioscorea trifida in the liquidizer together with yeast, oil and water.
was accomplished by comparisons with a voucher Then, this mixture was added to the previously mixed
herbarium specimen (Exsicata number 1353) deposited dry ingredients (wheat flour, powdered milk, sugar and
at the National Research Institute of Amazonia (INPA) salt). Bread manufacturing formulations can be seen at
Herbarium. It is very common to find the purple yam (Table 1). Homogenization (30 min.), dough underwent
(D. trifida) exhibiting several color hues of its flesh initial fermentation (60 min.), intermediate time for
A B
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) roxo (dark purple flesh);
D) branco (white flesh); and E) misto (white-purple flesh).

Table 1. Purple yam (D. trifida) incorporated
analyzed purple yam incorporated bread formulations (0%, 10%, 15% and 20%). Probability (P) values calculated from Kruskal-Wallis ANOVA followed
Table 3. Centesimal composition and caloric value mean and standard deviation on account of the dry material (except for the moisture content) of the four
bread formulations.
Caloric value (kcal/100 g)

Ingredients Type of bread*
294.45 15.63 a
269.17 21.82a
290.73 7.56a
280.64 3.95a
0% 10% 15% 20%
Wheat flour (g) 500 450 425 400
0.0862
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
Carbohydrate (%)
50.95 2.79a
53.41 4.07a
49.45 2.26a
53.43 3.13a
*
Percentage of wheat flour replaced by purple yam.

Total amount of ingredients used for preparing the
0.2587
breads.
bread shaping (25 min.), final fermentation (60 min.),
Values exhibiting different letters in the same column indicate statistically significant differences (P <0.05).
time for baking (30 min.) are followed for making yam
bread. After being prepared the breads were cooled to
1.18 0.28a
1.41 0.03a
1.38 0.07a
1.20 0.05a
room temperature and packed in polyethylene bags
Ash (%)
0.0683
displaying the products labeling.
Centesimal composition analyses of purple yam and
its incorporated breads
Crude Fiber (%)
Centesimal composition analyses of purple yam

1.95 0.08a
1.91 0,10a
1.84 0.07a
2.34 0.37a
(Dioscorea trifida), and purple yam incorporated breads
0.1438
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
11.62 0.81a
10.67 0,41a
10.06 2,12a
9.82 1,24a
described by the Instituto Adolfo Lutz-IAL (2008).

Protein (%)
Carbohydrate and caloric values were determined

0.2699
according to the method of (AOAC, 2005).
Findings obtained on the bread formulation
centesimal composition analyses were subjected to
4.49 0.12ab
4.87 0.37ab
4.74 0.10b
4.24 0,04a
Lipid (%)
0.0370
Table 2. Yam centesimal composition.

Yam Yam Purple Yam
Parameters
(D. spp.)* (D. alata)** (D.trifida)***
Moisture (%) 72.60 73.70 76.43 0.50
by post hoc tests are shown.
Lipid (%) 0.20 0.10 1.13 0.69

29.79 2.04a
31.03 0.83a
32.65 1.30a
35.09 5.62a
Moisture (%)
Protein (%) 2.00 2.30 1.83 0.13

Crude Fiber 0.60 7.30 1.80 0.05
0.0752
(%)
Ash (%) 0.90 0.90 0.78 0.02
Carbohydrate 24.30 23.00 18.04 0.66
(%)
Caloric value 100.00 96.00 89.64 4.52
Bread
10%
15%
20%
(Kcal/100g)
0%
*
(Montaldo, 1977), **(TACO 2006), ***Present study
statistical analysis through the statistical software (2004), with some modifications, where 2 mg of DPPH
package (Statsoft STATISTICA 8.0 2007). Given to the were dissolved into 15 mL of methanol, and applied so
number of sampled observations (n=3), Kruskal-Wallis as to determine the antioxidant activity of samples of
ANOVA and post hoc tests were applied as a purple yam and its incorporated breads in the four
non-parametric alternative to Fisher ANOVA, for aforementioned formulations. A micro plate bearing
independent data, in the comparison among the bread 96 well was used. Thirty microliters (30 L) of the
formulations. methanolic extract, plus 170 L of methanol (used as the
Findings showing significance level of (P<0.05) blank) were placed in the wells. The reading was
were considered as statistically significant. performed on an Elisa reader (DXL 800-BECKMAN
Preparation of purple yam and its incorporated COULTER) at a wavelength of 492 nm, using triplicate
breads methanolic extract samples. Then, 100 L of the DPPH solution were
Samples of purple yam (Dioscorea trifida), added, and the material was stored in a dark place for
were peeled and ground with the aid of a knife. They 30 min, and the reading was repeated as soon as this time
were then dehydrated in a laboratory oven at 60C for was over. Two hundred microliters (200 L) of methanol
24 h. Purple yam incorporated breads of four added to 100 l of the DPPH solution were used as
formulations: 0%, 10%, 15%, and 20%, were cut into the control. Thirty microliters (30 L) of quercetin
1 cm thick slices, and dehydrated in a laboratory oven at (10 g/mL), 170 L of methanol and 100 l of the
40C for 24 h (Hsu et al., 2004). Dehydrated yams and DPPH solution, were used as the standard. The following
breads were ground with pestle and mortar, weighed at formula was used so as to calculate the antioxidant
0.125, 0.25, 0.5 and 1.00 g (40, 80, 160 and 330 mg/mL, A sample - A blank
% AA = 100 - x 100
respectively). They were placed into small test tubes A control
added with 5 mL of methanol and left in a rotary shaker
for 24 h. The material was centrifuged at 2,500 RPM activity percentage
for 10 min so as to obtain the supernatant (methanolic Antioxidant activity determination through the lipid
extract). The antioxidant activity of the samples was peroxidation (LPO) method in purple yam and its
determined by the free radical scavenging, 2.2-diphenyl- incorporated breads
1-picryl-hydrazyl (DPPH) and lipid peroxidation (LPO) The determination of the antioxidant activity of
methods. The latter method evaluates the inhibition of the samples through the LPO method was carried out
free radicals generated during the linoleic acid according to the method reported by Duarte-Almeida
peroxidation, and is based on spectrophotometric et al., (2006), based on the methodology originally
measurements of discoloration (oxidation) of -carotene, described by Marco (1968), and later modified by Miller
induced by linoleic acid oxidative degradation products (1971). The reactive mixture was prepared in
(Marco, 1968; Miller, 1971; Duarte-Almeida et al., an Erlenmeyer flask, containing 50 L of linoleic acid,
2006). 200 L of tween 80 (emulsifying agent), 150 L of
Antioxidant activity determination through free -carotene solution at 2 mg/mL in chloroform, and
radicals scavenging methods (DPPH) in purple yam 500 L of chloroform. The mixture was then subjected to
and its incorporated breads evaporation in nitrogen till there was no more
DPPH method, following the methodologies chloroform left. Later, the mixture of 25 mL of
described by Shimada et al., (1992) and Hsu et al., previously oxygen saturated water was added, and during

a period of 30 min it was homogenized through vigorous software package (Statsoft STATISTICA 8.0 2007). The
shaking. Shapiro-Wilk test rejected the frequency distribution
The reactive mixture showed to be clear with normality of the three tested bread formulations, in all
absorbency ranging from 0.6 to 0.7 at a wavelength of their sensory attributes. However, the Levene test
492 nm. A 96 well bearing micro plate was used. Two accepted the homocedasticity (homogeneity of variances)
hundred forty microliters (240 L) of the reactive among the formulations for all sensory attributes. As
mixture and 10 L of the methanolic extract samples frequency distribution normality and variance
were placed in the wells. Ten microliters (10 L) of homogeneity are basic assumptions made for the
methanol and an equal volume of butylhydroxytoluene application of parametric tests, such as Fishers
(BHT) at a concentration of 40 g/mL were used as ANOVA, and as these assumptions were not attended to,
control and standard, respectively. The micro plate was the Friedman ANOVA followed by post hoc tests were
incubated at 50C to speed up the oxidation reactions and applied as a non-parametric alternative for paired data in
start -carotene discoloration. Discoloration slope bread comparisons. Findings presenting significance
readings of samples, control and BHT (in triplicate) were level of (P<0.05) were considered as statistically
performed readily, in an Elisa reader at a wavelength of significant.
492 nm every 15 min for 135 min. The following Microbiological analysis of purple yam breads
formula was used so as to calculate the oxidation Following the recommendation of the
A2 sample - A1 sample Brazilian National Health Surveillance Agency
% I = 100 - x 100 (in Portuguese, Agncia Nacional de Vigilncia
A2 control - A1 control
Sanitria, ANVISA), based on Ruling Number 12 (RDC,
inhibition percentage: 2001), we carried out the microbiological analysis so as
Sensory analysis of purple yam incorporated breads to verify Coliforms and Salmonella in samples of the
The acceptance test of purple yam in natura three purple yam bread formulation samples through the
incorporated breads counted with the participation of 78 membrane filtration method (APHA, 2001).
non-trained volunteer judges. Each one of them was
provided with an answering card bearing a 9 point RESULTS AND DISCUSSION
hedonic scale (9-like extremely to 1-dislike extremely), Centesimal composition and caloric value of purple
adapted from Stone et al., (1993) and Silva et al., (2005). yam
The judges were provided with three purple yam Moisture (76.430.50), protein (1.830.13) and
incorporated bread samples, produced from three ash (0.780.02) contents, as well as the caloric value
formulations (10%, 15% and 20%) (Table 1). Samples (89.644.52) of purple yam (D. trifida) samples analyzed
were served in white, disposable plastic plates; encoded in the present study (Table 2) show to be near
with three randomly chosen numbers. Samples were those presented by Montaldo (1991) for yam
evaluated according to their sensory qualities: global (Dioscorea spp.) and those found in the Brazilian
feel, aroma, flavor, color and texture. Judges were Food Composition Table TACO (2006), for the yam
advised to always rinse their mouth with water before (D. alata). Lipid content (1.130.69) stayed well above
testing the next sample. that presented by Montaldo (1991) and TACO (2006).
The findings obtained on the acceptance test Crude fiber content (1.800.05) is above the value
were submitted to statistical analysis through statistical observed by Montaldo (1991), and well below that
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).
Moisture Lipid Protein Crude Fiber Ash Carbohydrate Caloric value
Bread
(%) (%) (%) (%) (%) (%) (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
presented in TACO (2006). The high fiber content ordinary bread loaf (OBL) (Anton et al., 2006) and
presented by TACO (2006) might be due to the whole bread loaf (WBL) (TACO, 2006) (Table 4). One
enzymatic gravimetric method employed in the analyses. notices, a high fiber content (6.90%) in the whole bread
That method warrants a higher precision for determining loaf (WBL) (TACO, 2006), relative to the remaining
the dietary fiber as compared to the acid digestion breads. It can be highlighted that in whole bread
methodology used in the present study as well as composition, we have the presence of grain-composed
by Montaldo (1991). Total carbohydrate content whole flour, almost wholly made up of bran, germ and
(18.040.66) is well below Montaldo (1991) and TACO endosperm (FDA, 2006). By and large, all other values
(2006) values. The remaining differences in centesimal show to be approximate. All differences found may be
composition values presented by Montaldo (1991) and in related to formulations employed in the preparation of
the present study might be related to the different soil those breads.
types being employed on planting the tubers and/or to the Antioxidant activity determination through the free
different species being utilized. Nevertheless, the radical scavenging method (DPPH) in purple yam
different values presented in TACO (2006) may be and its incorporated breads
related to the different yam species being analyzed. Antioxidant activity (% AA) of the methanolic
Centesimal composition and caloric value of purple extract pertaining to purple yam (Dioscorea trifida)
yam incorporated breads
100
Based on data from Kruskal-Wallis (ANOVA)
90
followed by post hoc tests (Table 3), it may be asserted 80

Antioxidant activity (%)
70
that, except for the lipids (P<0.05), all other centesimal Purple yam
60 0% Bread
composition and caloric values of the four purple yam 50

10% Bread
15% Bread
40
incorporated bread formulations (0%, 10%, 15% and 20% Bread
Quercetin
30
20%) showed to be statistically similar (P>0.05). That is, 20
10
replacing wheat flour by purple yam in natura in up to
0
20% neither modifies bread centesimal composition nor 40 80 160 330

Concentration (mg/mL)
caloric value. As for lipid, statistically significant

Figure 2. Antioxidant activity expressed by free
difference was only observed for 10% and 20%
radical scavenging percentage, of samples of purple
formulations; this negligible 0.5% difference may be yam (Dioscorea trifida) and its incorporated bread
extracts in four formulations: 0%, 10%, 15% and
neglected in technological applications.
20%, as determined by DPPH method. The quercetin
Purple yam incorporated breads centesimal was used as standard control. Bars indicate standard
deviation.
composition and caloric value were compared to those of

samples in the concentrations of 330, 160, 80 and LPO method confirmed the antioxidant activity (% I) in
40 mg/mL, as determined by the DPPH method, were purple yam (55.804.85) and its breads from the three
higher than 70%, reaching a maximum of 88.130.12. formulations (10%, 15% and 20%), with the values of
This plainly shows this species to exert DPPH radical 46.164.90; 48.203.72 and 49.132.79, respectively.
scavenging activity (Figure 2). This same figure reveals
purple yam incorporated breads prepared in 10%, 15% 0.9
0.8
and 20% formulations, to also present a certain
0.7 Blank
antioxidant activity, reaching 43.321.18; 48.131.17 0.6 Purple yam
0% Bread
and 53.711.01 maximum percentile values,
Abs 492
0.5 10% Bread
0.4 15% Bread

respectively. Those findings are above the values 0.3
20% Bread
BHT
presented by Hsu et al., (2004) (20-40% approximately), 0.2
0.1
who used breads of several formulations prepared with
0
flour from the purple yam tuber (Dioscorea purpurea) 0 15 30 45 60 75 90 105 120 135
Time (min)
representing the one with the widest variety in Taiwan,
for substituting part of the wheat flour. Bread prepared Figure 3. Discoloration slope of purple yam
(Dioscorea trifida) and its incorporated bread
with no purple yam at all (0%) showed certain extracts in four formulations: 0%, 10%, 15%, 20%,
antioxidant activity, as well, probably due to Maillard blank and BHT, as determined through the LPO
method.
reaction products, where, some hot processed foods,
present free radical scavenging activity (Kim et al.,
90
2007; Jing and Kitts, 2000; Hsu et al., 2004; Michalska 80
et al., 2008). Corroborating data from Hsu et al., (2004), 70
it was confirmed that the antioxidant activity rose as the 60

Inhibition (%)
50
percentage of purple yam substituting wheat flour
40
increased. The high free radical scavenging activity 30
observed by Hsu et al., (2004) in flour of Taiwan purple 20
yam (D. purpurea), was also detected in the Amazonian 10
0
regions purple yam (D. trifida). Purple yam 0% Bread 10% Bread 15% Bread 20% Bread BHT
Antioxidant activity determination through the lipid

Figure 4. Inhibition percentage of free radicals of
per oxidation (LPO) method in purple yam and its purple yam (Dioscorea trifida) and its incorporated
bread extracts in four formulations: 0%, 10%, 15%,
incorporated breads
20%, and BHT as determined by LPO method. Bars
Discoloration slope (Figure 3) and free radical indicate the standard deviations.
inhibition activity (Figure 4) determined through the
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.
Tests Bread% Color Aroma Flavor Texture Overall impression
10 0.0009 < 0.0001 0.0023 0.0044 0.0001
Shapiro-Wilk P 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
Values were considered statistically significant at (P< 0.05).
As it was observed by the DPPH method, the
Median
antioxidant activity rose as the percentage of purple yam
Overall impression
7
substituting wheat flour in the breads increased.
0.0608
Moreover, bread with no addition of purple yam (0%)
6.54 1.23
6.59 1.14
6.88 1.33
presented some antioxidant ability which might have
Mean
resulted from the development of Maillard reaction
products (Kim et al., 2007; Jing and Kitts, 2000; Hsu
Table 6. Sensory evaluation results of the three purple yam incorporated bread formulations. Probability (P)
et al., 2004; Michalska et al., 2008). Anthocyanins might
Median
be partly responsible for the antioxidant activities
6
detected in the purple yam (D. trifida) and its
Texture
incorporated breads analyzed in the present study, since
0.5264
value was obtained through Friedman ANOVA followed by post hoc test.
these pigments were detected in purple yams, D. alata
6.03 1.50
5.94 1.67
6.22 1.61
Mean
(Rasper and Coursey, 1967) and D. trifida L.
Values exhibiting different letters in the same column point out statistically significant differences (P< 0.05).
(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
Median
antioxidant activity in yams (Hou et al., 2001; Hsu et al.,
6
2004). Flavor
0.9641
Sensory analysis of purple yam incorporated breads
6.28 1.44
6.44 1.22
6.41 1.43
Table 5 shows the rejection of the frequency Mean
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
Median
homocedasticity among the formulations through the

6,5
6
Levene test on all sensory attributes evaluated in the

Aroma
acceptance test (i.e. statistically significant values at 0.6285

6.71 1.22
6.51 1.34
6.68 1.20
(P <0.05).
Mean
Friedman ANOVA followed by post hoc tests

applied for comparing the three purple yam incorporated
bread formulations revealed a significant difference
Median
(P<0.05), only for the colour attribute (Table 6). The

6
bread at 20% presented a better evaluation regarding the

0.0001
Color
remaining ones, probably due to the higher purple yam

6.29 1.30ab
6.97 1.55b
6.15 1.46
concentration, which gives the final product a more

Mean
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
Bread
10%
15%
20%
6.15 to 6.97).
P
Choosing a determined food should depend

Table 7. Purple yam incorporated breads microbiological analysis.
Type of bread
Microorganism
10% 15% 20% Based on RDC (2001)*
Coliforms (at 45C/g) Absent Absent Absent 102
Salmonella sp/25 g Absent Absent Absent Absent
*Ruling Number 12 (RDC 2001) recommended by the Brazilian National Health Surveillance Agency
(in Portuguese, Agncia Nacional de Vigilncia Sanitria - ANVISA).
mainly on its nutritional value. Nevertheless, color, ACKNOWLEDGEMENTS

aroma and texture are the factors usually guiding the The authors are indebted to the Fundao de
consumers preference rate. Of these three factors, color Amparo Pesquisa do Estado do Amazonas (FAPEAM)
interferes the most on the products preference (Bobbio for the master scholarship granted to Antonia Paiva
and Bobbio, 2001). Teixeira. To Dr. Antonio Jos Inhamuns da Silva and
Given that there were no preferential differences MSc. Cynthia Tereza Corra da Silva of Universidade
among the other sensory attributes, the three breads Federal do Amazonas (UFAM) for kindly having
evaluated can be considered approved. allowed to carry out the centesimal composition analyses
Microbiological analysis in their laboratories. To MSc. Antonio Fbio Lopes de
Considering that the microbiological analysis Sousa, Arleilson de Sousa Lima and Ana Cludia dos
was negative for Coliforms and Salmonella (Table 7), the Santos for their invaluable aid in undertaking of the
purple yam incorporated breads may be considered laboratory analyses. To Misters Claudio Adriano
proper for human consumption, as long as they have Cardoso Amanajs and Francisco de Oliveira Batista for
been properly handled. their logistical support in the collection of purple yam
samples.
CONCLUSIONS
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Pedralli G. 1988. O inhame, esse desconhecido.
Cinc Hoje 8(46):58-62. Advantages
Pedralli, G. 1997. Reviso taxonmica das espcies de Complete Peer review
Affordable Charges
Dioscoreaceae (R.Br.) Lindley da Cadeia do Espinhao, Quick processing
Minas Gerais e Bahia. Tese doutorado PG - Botnica, Extensive indexing
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Original Research
Bioefficacy of Novaluron, a chitin synthesis inhibitor against the tropical

warehouse moth, Ephestia cautella.
Authors: ABSTRACT:
Sackey I, Eziah VY and The tropical warehouse moth, Ephestia cautella (Lepidoptera: Pyralidae) is a
Obeng-Ofori D. 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
Institution: ranged between 50-80% after 96 h of exposure. Also, Novaluron (0.5 mL/L) was used
Department of Crop Science, to treat four surfaces (concrete, wood, glass and plastic) usually encountered in
College of Agriculture and structural insect pest management systems and the larvae exposed to these surfaces.
Consumer Sciences, P. O. Hocklicombi (5 mL/L) served as positive control. Larval mortality (35.5-97.5%),
Box LG 44, University of pupation (0.0-35.0%) and adult emergence (0.0-20.0%) in surfaces treated with
Ghana, Legon. 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%).
Corresponding author: Keywords:

Eziah VY. Novaluron, Hocklicombi, Ephestia cautella, warehouse moth, chitin,
loss assessment.
Web Address: Article Citation:

http://jresearchbiology.com/ Sackey I, Eziah VY and Obeng-Ofori D.
documents/RA0305.pdf. Bioefficacy of Novaluron, a chitin synthesis inhibitor against the tropical warehouse
moth, Ephestia cautella.
Dates:
Received: 08 Nov 2012 Accepted: 27 Nov 2012 Published: 17 Jan 2013
cited.
759-767 | JRB | 2013 | Vol 3 | No 1

Sackey et al., 2013
INTRODUCTION laboratory. There is no information on the use of

Maize (Zea mays) is one of the major staple food novaluron for stored product protection in warehouses or
crops in Ghana and it is susceptible to attack by several cribs. Ephestia cautella is noted for feeding directly on
insect pests including the tropical warehouse moth, the grains and also, the mature larvae leave the
Ephestia cautella Walker (Lepidoptera: Pyralidae) commodity in search of pupation sites in crevices, cracks
(CAB1, 2006). Ephestia cautella larva feeds on stored and storage containers. Therefore, treating these surfaces
products, damaging the product directly and form webs to which the insect may be exposed will go a long way to
on the surface. The webbing contains larval excreta and mitigate the losses caused by this pest. Hence, screening
exuviae which give unpleasant odour to the infested Novaluron against E. cautella using the commodity and
commodity. Older larvae may leave the food to find storage surfaces as substrates is crucial in the
pupation sites in wall cracks. management of the pest. Such findings will contribute to
In Ghana, E. cautella is controlled by the use of the efforts by farmers and warehouse managers to reduce
residual insecticides usually, synthetic pyrethroids and storage losses and contribute to the attainment of food
fumigants (CABI, 2006). The adverse effects of residual security in Ghana.
pesticides such as poisoning, environmental and health This study presents laboratory and field tests that
hazards and resistance development cannot be were carried out to determine the toxicity of novaluron to
overemphasized (Obeng-Ofori, 2007). Hence the use of the 5th instar larvae of E. cautella. Other tests were also
residual insecticides in stored product protection is conducted to assess the efficacy of novaluron on
challenging. There is therefore, the need for new cost and different surfaces against immature stages of E. cautella.
environment friendly alternatives with no adverse effect
on non-target organisms (Obeng-Ofori, 2007; Arthur and MATERIALS AND METHODS
Phillips, 2003). Some of these alternatives include Insect cultures
botanicals, insect growth regulators, microbial pathogens The test insects were obtained from the
among others (Arthur, 1996). Entomology laboratory of the Crop Science Department.

Novaluron (Rimon 10 EC) is a benzoylphenyl Adult E. cautella were cultured on mixed substrate made
urea group of insect growth regulators and a chitin up of wheat powder, maize flour and glycerol (5:5:1).
synthesis inhibitor. Novaluron has been registered as an Fifty adult E. cautella were introduced into each jar and
insecticide for food crops in several countries including left under laboratory conditions of 272C and 55-60%
South Africa, Australia and Ghana (WHO, 2003; EPA, relative humidity for 30 days to allow for the
2006). In Ghana, novaluron has been successfully used development of larval E. cautella. The set up was placed
in the laboratory against stored product pests on trays containing industrial oil to prevent the crawling
such as the rice moth, Corcyra cephalonica of other insects into the culture. The insects were reared
Stainton (Lepidoptera: Pyralidae) (Sarbah, 2006), the and handled using ethically acceptable standard
red flour beetle, Tribolium castaneum Herbst procedures in the laboratory.
(Coleoptera: Tenebrionidae) (Bakudie, 2006) and the Test chemicals
tropical warehouse moth, E. cautella (Ibrahim, 2008). Novaluron (Rimon 10EC), 1-[3-chloro-4-(1, 1,
In Ghana, most of the work done on novaluron 2-trifluoro-2-trifluoromethoxy-ethoxy) phenyl]-3-(2, 6
focused on evaluating the effect of the chemical on difluorobenzoyl) urea, produced by Makhteshim-Agan
different developmental stages of insects in the Ltd (Israel) was used for the toxicity experiment and
Sackey et al., 2013
Hocklicombi (Hockley International Ltd. Poynton, dishes were used as glass surfaces for the surface
Stockport, U. K.) which contains 25% Fenitrothion and treatment.
5% Fenvalerate was used as a reference product. Each of the four surfaces was treated with 4 mL
Contact toxicity test of water (negative control treatment) or an aqueous
We adopted the method by (Eziah et al., 2011). solution of novaluron (0.5 mL/L) and Hocklicombi
Concentrations of Novaluron (0.1, 0.2, 0.3, 0.4 and (5 mL/L) (positive control). All treated arenas were

0.5 mL/L) and 5 mL/L of Hocklicombi were diluted in allowed to dry overnight and fifth instar larvae (N=10) of
distilled water and used for the assays. Distilled water E. cautella were exposed for 48 h. The larvae were then
was used as negative control. Fifth instar larvae of both transferred to new petri dishes containing food under
sexes were transferred into clean Petri dishes and the laboratory conditions of 272C and 55-60% relative
different dosages of the various concentrations was humidity. Post-treatment survival and mortality were
topically (1 L) applied to the notal regions of the larvae recorded daily. Number of surviving larvae that
using a micro applicator. Each experimental unit successfully pupated and those that successfully emerged
consisted of 10 larvae and was replicated for four times. as adults were recorded.
The treated insect larvae were then transferred into glass Field experiment
petri dish containing food. The insect larvae were Maize grains were obtained from the Madina
examined for mortality 24, 48, 72, 96 h, 7 days and (a suburb of Accra, Ghana) market and sieved to remove
14 days after treatment. Criterion for death was as all debris. Maize grains (5 kg) were sterilized in the oven
described by (Lloyd, 1969) in which insects were at 70C for 3 h after which they were left in desiccators
presumed dead when they failed to move in a to cool. The grains were then treated with 0.4 mL/L
coordinated manner after prodding with a blunt probe. Novaluron or 5 mL/L Hocklicombi. These dosages had
Data collected include larval mortality, percent pupation proven effective in laboratory experiments. Grains
and percent adult emergence were done after various treated with distilled water served as negative control.
treatments and exposure periods. Each treatment was replicated four times. Fifty unsexed
Surface treatment adults of E. cautella were put onto the treated grains in
The surfaces chosen for the study were concrete, each sack. The sacks were securely sealed by stitching
plywood, glass and plastic which are among the and stored in a grain crib at the University farm for
common surfaces encountered in structural insect pest 60 days. Prior to their treatment, subsamples were taken
management. Individual concrete exposure arenas were from each sack for moisture and weight loss analyses
created in square bottoms of plastic containers (6x6 cm) using the standard volume method was carried out
using a concrete patching material. Water-based slurry (Boxall, 1986). At the end of the storage period, the
was prepared by mixing 1 kg of Portland cement to 2 kg contents of the sacks were sieved. The number of
of sand and 1 L of tap water and pouring 10 mL of the both live and dead adult insects was recorded. Also,
slurry into the bottom of the plastic container to create a subsamples of the maize grains were collected for
treatment arena (Arthur, 1998b). Plywood arenas were moisture and weight loss analyses as stated earlier.
made by cutting rectangular disks from 1.25 cm thick Statistical analysis
plywood to fit the plastic container then caulking the Data involving percentages were arcsine
margins to prevent the larvae from escaping the surface. transformed and were analyzed using the Analysis
Plastic containers served as plastic surfaces and petri of Variance (ANOVA) with Genstat 9.2

Sackey et al., 2013
(Lawes Agricultural Trust, 2007). Means were separated different from the negative control (64.0-80.0%)
using the Least Significant Difference (LSD) test at (Figure 1). However, all other concentrations of
5% probability level. Novaluron higher than 0.1 mL/L significantly (p = 0.05)
impaired pupation. There was no significant difference in
RESULTS pupation 7 days after the exposure of E. cautella larvae
Contact toxicity test to 0.5 mL/L Novaluron and 5 mL/L Hocklicombi.
The percent larval E. cautella mortality Also, after 14 days, percentage pupation recorded in

following treatment with Novaluron and Hocklicombi larvae treated with 0.4 mL/L and 0.5 ml/L was
are presented in Table-1. Larval mortality varied with comparable.
insecticide concentration and exposure period. Lower All levels of Novaluron concentrations
dosages of Novaluron (0.1-0.3 mL/L) caused less significantly reduced the development of F1 of adult
than 50% larval mortality after 96 h of exposure E. cautella (Figure 2). The highest adult emergence
(Table 1). In contrast, novaluron concentrations of (77.5%) was recorded in the negative control and this
0.4 mL/L and 0.5 mL/L caused between 50% to 80% differed significantly (p = 0.05) from all other novaluron
larval mortality after 72 to 96 h of exposure. After 96 h concentrations applied. As concentration increased from
exposure period, all dosages of Novaluron induced 0.1 mL/L to 0.5 mL/L, adult emergence significantly
significantly (p = 0.05) higher larval mortality compared (p = 0.05) reduced from 50 to 2.5%. Also, the effect of
to the negative control. However, there was no novaluron applied at 0.5 mL/L was comparable to
significant difference in larval mortality between 5 mL/L Hocklicombi treatment in impairing the development of
Hocklicombi and 0.5 mL/L Novaluron treatments. Also, adult E. cautella.
novaluron applied at 0.4 ml/L and 0.5 mL/L did not Surface treatment
differ significantly from each other after 96 h of Ephestia cautella larvae exposed on concrete
exposure. surfaces treated with Novaluron showed a lower
Pupation and adult emergence of E. cautella mortality than those exposed to concrete surfaces treated
were observed in all insecticide treatments and the with Hocklicombi (Table 2). Mortality was also lower
negative control with the exception of Hocklicombi on plastic and wood treated surfaces compared to
treatment. The percentage pupation in larvae treated with Hocklicombi treated surfaces. However, the percentage
0.1 mL/L Novaluron (57.5-65.0%) was not significantly mortality of E. cautella on glass surfaces treated with
Table 1 Mortality of larval E. cautella (%) after treatment
with novaluron and Hocklicombi insecticides
Treatments (ml/L) Mean(s.e) % larval mortality (h)
24 h 48 h 72 h 96 h
Control (Water) 0.00.0 0.00.0 0.00.0 0.00.0
5.0 mL/L (HC) 87.50.1 87.50.1 87.50.1 87.50.1
Novaluron
0.1 7.50.0 10.00.0 17.50.1 22.50.1
0.2 10.00.0 12.50.0 27.50.1 42.50.1
0.3 17.50.1 25.00.1 35.00.1 45.00.2
0.4 17.50.1 32.50.1 50.00.1 66.00.1
0.5 32.50.1 47.50.1 65.00.1 80.00.0
LSD (P < 0.05) 18.35 18.40 14.50 14.83
HC= Hocklicombi
s.e = standard error

Sackey et al., 2013
Table 2 Mortality of E. cautella larvae (%) after 7 days exposure on concrete, glass,
plastic and wood surfaces treated with Hocklicombi and novaluron insecticides
Mean (%) s.e mortality
Insecticide Type of surface
Concrete Glass Plastic Wood Means
Control 0.0 0.0 0.00.0 2.5 0.0 0.0 0.0 0.70.0
Hocklicombi 43.0 0.1 97.50.0 45.0 0.1 35.0 0.1 55.00.0
Novaluron 25.00.1 97.50.0 17.5 0.1 25.0 0.1 41.10.0
Means 22.7 0.0 64.70.0 21.7 0.0 20.0 0.0 -
LSD(P < 0.05): Main effects (insecticide = 1.21, surface= 1.39 Interaction (insecticide x surface)=2.4
novaluron was the same (97.5%) as those treated with which were not treated (11.3%) using standard volume
Hocklicombi. Surviving larvae were observed for methods.
pupation and adult emergence. Fewer E. cautella larvae DISCUSSION
pupated after exposure to concrete, plastic and wood The present study showed that Novaluron

surfaces treated with Hocklicombi but no pupation was concentrations of 0.4 mL/L and 0.5 mL/L significantly

recorded on glass surfaces treated with Hocklicombi affected the metamorphosis of E. cautella to the adult
(Table 3) stage. The effectiveness of novaluron at these dosages
Fewer larvae pupated in glass surfaces-treated compared favourably with Hocklicombi . The insect
with novaluron and this was not significantly different growth regulators ability to regulate metamorphosis in
from Hocklicombi-treated glass surfaces. Generally, the larvae through contact by topical application is
percentage adult E. cautella that emerged was greater on consistent with its mode of action. Tomlin (2005)
the untreated control for all the surfaces and differed reported that novaluron was very effective on the larvae
significantly (p = 0.05) from all insecticide treated of insects when absorbed by ingestion and contact
surfaces (Table 4). Mean percentage adult emergence of activity. The author also reported that the compound
E. cautela observed on glass and plastic surfaces treated causes abnormal endocuticular deposition and abortive

with novaluron and Hocklicombi ranged from 0.0 to moulting.
25%. Thus, residual effects of novaluron and Although pupation and adult emergence were

Hocklicombi significantly reduced the development of observed in all treatment levels, most of the larvae
E. cautella on glass and plastic surfaces. treated with 0.4 mL/L and 0.5 mL/L Novaluron could not
Field experiment emerge into adults 23 days after treatment. This may be
Table 5 shows the dry weight loss of the treated attributed to abnormal endocuticular deposition and
grains after 60 days of storage using the standard volume abortive moulting in the larvae (Tomlin, 2005). Also,
method. Lower weight losses were observed in grains when cocoon covering the pupae were slightly removed,
treated with insecticides (6.6-6.8%) compared to grains pupae found were malformed compared to those in the
Table 3: Percentage pupation of E. cautella after 14 days exposure on concrete, glass,
plastic and wood surfaces treated with Hocklicombi and novaluron insecticides
Means (%) s.e for pupation
Control 92.50.0 95.00.0 95.00.0 97.50.0 95.00.0
Hocklicombi 35.00.1 0.00.0 35.00.1 25.00.0 23.80.0
Novaluron 55.00.1 2.50.0 60.00.1 47.50.1 43.10.0
Means 61.90.0 41.20.0 63.10.0 56.20.0 -
LSD(P < 0.05): Main effects (insecticide5.6, surface= 6.6) Interaction (insecticide x surface)=13.20

Sackey et al., 2013
control. Adults that emerged were found not to be active development stage, time of application, kind of
as those in the control. These findings are consistent with compound and dose administered.
reports by Amos and Williams (1974). According to The residual effect of Hocklicombi and
CABI (2006), pupal formation is completed in seven Novaluron were significantly greater on glass surfaces
days and development from egg to adult ranges from than plastic, concrete or wood surfaces. Generally,
o
29-31 days under optimum conditions of 32.5 C and Hockicombi significantly caused higher mortalities on
70% relative humidity. However, in the present study all the surfaces than novaluron. The high residual
under laboratory conditions of 272C and 55-60% efficacy of Hocklicombi may be attributed to the
relative humidity, pupation extended up to 14 days and components of the compound. Hocklicombi contains
adult emergence was also delayed up to 30 days in the fenitrothion and fenvalerate as its active ingredients.
th
treated 5 instar larvae of E. cautella. Thus, novaluron These compounds have been reported by several
was found to prolong the development period of researchers to have high residual effects when used as
E. cautella larvae to adults. surface treatment against storage insects (Orui, 2004).
The ability of Novaluron to reduce the number of Both compounds are non-systemic insecticides with
new generations is consistent with the findings of contact and stomach activity (Tomlin, 2005).
(Kostyukovsky et al., 2003) and Kostyukovsky and In the present study, novaluron demonstrated
Trostanetsky (2006). The authors found that novaluron excellent residual effect on glass surfaces by preventing
applied at 1 ppm reduced the number of new generations the metamorphosis of E. cautella to the adult stage. The
of S. oryzae and R. dominica by 95% and also caused residual effect on glass surfaces treated with novaluron
rd
total mortality of the 3 instar larvae of T. castaneum. compared well with Hocklicombi. However, on plastic,
The effectiveness of novaluron in preventing the concrete and wood surfaces, Novaluron was less
metamorphosis of E. cautella when applied at 0.4 mL/L effective compared with Hocklicombi but differed
and 0.5 mL/L also confirms work done by Ibrahim significantly from the untreated control surfaces.
(2008). The author found that development of E. cautella However, the residual effectiveness on plastic surfaces
to adults was prevented when novaluron was applied at showed better efficacy than on concrete and wood
0.4 mL/L and 0.6 mL/L. surfaces.
These observations indicate that the effectiveness The excellent effectiveness of Novaluron on
of novaluron as a grain protectant depends on the species glass and plastic surfaces is consistent with work done by
of insect, dosage and exposure time. Wilson and Cryan (Atkinson et al., 1992). The authors found that when
(1997) and Mulla et al., (2003) stated that the effects of hydropene, an insect growth regulator was sprayed on
chitin synthesis inhibitors vary according to species, non-absorbent surfaces such as glass and ceramic tile, the
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
Means (%) s.e for adult emergence
Control 92.50.0 90.00.0 95.00.0 97.50.0 93.80.0
Hocklicombi 20.00.1 0.00.0 12.50.1 12.50.0 11.20.0
Novaluron 42.50.1 0.00.0 25.00.1 42.50.1 27.50.0
Means 50.60.0 32.50.0 45.00.0 48.10.0
LSD(P < 0.05): Main effects (insecticide5.9, surface= 6.34)= 6.34 Interaction Insecticide x surface=12.67
Sackey et al., 2013
Figure 1 Percentage pupation (meanss.e) of Figure 2 Percentage adult emergence (meanss.e) of

E. cautella larvae after treatment with novaluron and E. cautella after treatment with novaluron and
Hocklicombi insecticides. d = days h= hours Hocklicombi insecticides. d = days
survival, number of oothecae and percentage of grains compared to the control. Novaluron was observed
cockroaches were more affected than on absorbent to significantly reduce insect numbers in the treated
surfaces of finished plywood and fibreboard. The low grains and also had a significantly lower dry weight loss.
mortality rates, pupation and adult E. cautella that Results from this study showed that novaluron
emerged after exposure to concrete and wood surfaces in effectively protected maize grains from damage by
the current study can also be attributed to the E. cautella. Grain weight losses calculated in the
composition of these surfaces. Burkholder and Dicke Novaluron treatment compared well with those observed
(1966) reported that new concrete surfaces contain high in grains treated with Hocklicombi. Considering that
levels of alkaline which hydrolyze residues and reduce Novaluron selectively targets larval stages by inhibiting
residual efficacy of insecticides hence, the low mortality chitin synthesis and therefore, minimizes its impact on
rates on concrete-treated surface in the present study was adults of non targeted insect species (Ishaaya et al.,
not unexpected. Chadwick (1985) attributed low efficacy 2001), Novaluron can be used in replacement of residual
of insecticides on plywood surfaces to vaporization, insecticides like Hocklicombi for treatment of maize
chemical degradation, photodegradation and absorption grains for storage.
of insecticides into surfaces. Thus, the low mortalities
and higher survival rates observed in E. cautella exposed CONCLUSION
to wood surfaces treated with the insecticides may be due The current study showed that Novaluron was
to the absorption of the insecticide into the wood effective in controlling the tropical warehouse moth. The
surfaces after treatment. application of Nuvaluron at 0.4 mL/L and 0.5 mL/L
In the field experiment, all the insecticide treatments resulted in larval mortality ranging between
treatments significantly reduced dry weight loss in the 50-80% after 96 h of exposure. Also, the treatment of
Table 5 Percent dry weight loss after 60 days of concrete, wood, glass and plastic surfaces usually
storage using the standard volume method encountered in structural insect pest management
Dosage (mL/L) Mean dry weight loss (%) systems with 0.5 mL/L Novaluron induced (25.0-97.5%)
Control 11.30.0 larval mortality, (2.5-60%) pupation and ((0.0-42.5%)
Hocklicombi 5 6.60.0
Novaluron 0.4 6.80.0 adult emergence. These figures were comparable to
LSD(P < 0.05) = 1.63 those obtained from surfaces treated with 5 mL/L

Sackey et al., 2013
Hocklicombi insecticide. In the field maize treated with Bakudie E. 2006. Susceptibility of Tribolium castaneum
0.4 mL/L Novaluron and infested with adult E. cautella to novaluron on maize and rice. Bachelor of Science
after 60 days of storage showed that there was a lower Dissertation. Department of Crop Science, University of

weight loss in the Hocklicombi (6.6%) and novaluron Ghana, Legon, 36.
(6.8%) treatments compared to the negative control
Boxall RA. 1986. A critical review of the methodology
(11.3%). This work has proven that Novaluron could
for assessing farm grain losses after harvest. Tropical
replace the synthetic insecticides that are used in the
Development and Research Institute Report G191, Viii
management of this pest and should be included in the
139.
management programmes for storage pests control.
Burkholder WE and Dicke RJ. 1966. The toxicity of
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Complete Peer review
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Extensive indexing
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efficacy of insecticide on the cigarette bettle,
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Original Research
A Checklist of Butterflies of Meenachil River Basin, Kerala, India

Authors: ABSTRACT:
Vincy MV1, Brilliant R2
and Pradeepkumar AP3
Institution: Butterflies are highly sensitive to environmental change and are delicate
1. School of Environmental creatures that act as good bio-indicators of the health of an ecosystem. Meenachil
Science, Mahatma Gandhi river basin has attracted considerable amount of public interest. A survey of the
University, Kottayam,
butterflies conducted randomly revealed a total of 91 species belonging to five
Kerala.
families including three endemic species. Family Nymphalidae dominated in the study
2. PG Department of area, followed by Hesperiidae and Lycaenidae. This area is currently under severe
Environmental Sciences, anthropogenic pressure and minimizing these disturbances is important for the
St. Johns College, Anchal, long-term survival of specialist butterflies.
Kerala.
3. Department of Geology,
University of Kerala,
Kariavattom, Kerala.

Vincy MV. Meenachil river, Endemic species, bio-indicators, anthropogenic pressure.
Email: Article Citation:

vincybrilliant@gmail.com Vincy MV, Brilliant R and Pradeepkumar AP.
A Checklist of Butterflies of Meenachil River Basin, Kerala, India.
Web Address:
http://jresearchbiology.com/ Dates:
documents/RA0308.pdf. Received: 21 Nov 2012 Accepted: 03 Dec 2012 Published: 04 Feb 2013
cited.
768-774 | JRB | 2013 | Vol 3 | No 1

Vincy et al., 2013
INTRODUCTION Meenachil river basin in Kerala, South India. However,

Butterflies are the most beautiful and colourful comprehensive long-term ecological studies to monitor
creatures on the earth and have a great aesthetic value. the butterfly population of the area remains as a serious
India harbours about 1501 species of butterflies lacuna. Such studies are imperative to improve the
(Haribal, 1992), 285 species are found in southern India ecological utility of butterflies as indicator taxa.
(Thomas, 1966), of which 45 species are endemic to
southern India. Butterflies, widely appreciated for the MATERIALS AND METHODS
aesthetic value are important as ecological indicators The present study is an attempt to provide a
(Chakravarthy et al., 1997) and flagship taxa in checklist of butterflies based on a four-year field study
biodiversity inventories (Lawton et al., 1998). from October 2008 to October 2012. Identification of
Meenachil river which is one of the important species was done using available literature (Evans, 1932;
river of Kottayam district in Kerala, emerges from Gunathilagaraj et al., 1998; Haribal, 1992; Palot et al.,
Western Ghats and confluences into Vembanad Lake. 2003; Gay et al., 1992; Wynter-Blyth, 1957) and with
This river has a total length of 78 km and has the help of experts. Species classification and scientific
a catchment area of 1272 km2. The entire Meenachil names are as per Gunathilagaraj et al., (1998).
watershed area geographically lies between 925 N
to 955 N latitude and 7630 E to 7700 E longitude. RESULT AND DISCUSSION
The general elevation of the entire river basin ranges The study during the period indicate that the
from 77 m to 1156 m in the high lands and less than 2 m habitats where butterflies were found and captured are
in the low lands. The Meenachil river basin falls within disturbed areas and are strongly influenced by
the realm of tropical climate. The temperature of the area anthropogenic activities. These range from city lots to
varies in between 24C and 32C throughout the year. pasture, abandoned fields, road sides, plantations,
The annual rainfall varies from less than 100 cm to more riparian area, etc.
than 500 cm with an average of 300 cm. The occasional A total of 91 species belonging to 71 genera
rainfall is also received between the two seasons. Rubber distributed over five families were collected from the
trees are extensively cultivated in vast areas in the entire monitoring sites, during the study period. The family
river basin. Besides rubber, other crops like spices, Nymphalidae dominated with 34 species followed
paddies etc., are also cultivated in the river basin area by Hesperiidae (20 spp.), Lycaenidae (18 spp.), Pieridae
(Watershed Atlas, 1996). (7 spp.), and Papilionidae (12 spp.). Even though, the
Among insects, butterflies are the most family Nymphalidae exhibited the maximum species
studied group. Larsen (1987a, b, c, 1988) made a detailed diversity, family Pieridae showed maximum species
survey of butterflies of Nilgiri Mountains and recorded density. Three butterfly species recorded from this region
n earl y 300 speci es in cl udin g en demi cs. have protected status under the Wildlife Protection Act,
In Kerala, documentation of butterflies on Silent Valley 1972 (Arora, 2003). They are Hypolimnas misippus and
National Park (Mathew and Rahamathulla, 1993) Atrophaneura hector included under Schedule I Part IV
an d Par a m bi kula m Wi ld li fe San ctuar y and one species Aeromachus pygmaeus in Schedule II
(Sudeendrakumar et al., 2000) have been carried out. Part II. Further research with reference to ecology,
The present paper presents a checklist and diversity of threats and conservation of butterflies in the area is in
butterfly populations in different altitude levels in progress.
Table 1. List of butterfly species collected in the study areas and the plants visited by them
Sl. No. Common Name Scientific Name Wingspan Status Plants visited
FAMILY PAPILIONIDAE
1 Common Blue Bottle Graphium sarpedon (Linnaeus) 80-90 mm Common Litsea chinensis, Polyalthia longifolia,
Cinnamomum malabatrum, Persea
Vincy et al., 2013
odoratissima, P. macrantha
2 Tailed Jay Graphium agamemnon (Linnaeus) 85-100 mm Common Polyalthia longifolia, Cinnamomum spp.,
Annona reticulata, A. squamosa
3 Common Mime Papilio clytia (Linnaeus) 90-100 mm Common Litsea chinensis
4 Common Mormon Papilio polytes (Linnaeus) 90-100 mm Very Common Citrus spp., Glycosmis arborea, Murraya
koenigii, curry leaf plant
5 Blue Mormon Papilio polymnestor (Cramer) 120-150 mm Not rare Citrus limona, Glycosmis arborea
6 Lime Butterfly Papilio demoleus (Linnaeus) 80-100 mm Very Common Glycosmis arborea, Murraya koenigii
7 Paris Peacock Papilio paris (Linnaeus) Citrus spp.
8 Common Rose Atrophaneura aristolochiae (Fabricius) 80-110 mm Common Thottea siliquosa
9 Malabar Rose Atrophaneura pandiyana (Moore) 100-130 mm Common Thottea siliquosa

10 Crimson Rose* Atrophaneura hector (Linnaeus) 90-110 mm Common Aristolochia indica, Thottea siliquosa
11 Common Birdwing Troides helena (Linnaeus) 140-170 mm Not rare
12 Southern Birdwing Troides minos (Cramer) 140-190 mm Not rare Aristolochia indica, Thottea siliquosa
FAMILY PIERIDAE
13 Common Grass Yellow Eurema hecabe (Linnaeus) 40-50 mm Common Caesalpinia spp., Cassia tora, C. fistula,
Acacia spp.
14 Common Emigrant Catopsilia pomona (Fabricius) 55-80 mm Common Bauhinia racemosa, C. fistula, C. tora,
Butea monosperma
15 Mottled Emigrant Catopsilia pyranthe (Linnaeus) 50-70 mm Common C. fistula, C. tora
16 Great Orange Tip Hebomoia glaucippe (Linnaeus) 80-100 mm Common Capparis spp.
17 Chocolate Albatross Appias lyncida (Cramer) 55-70 mm Common Capparis spp.
18 Common Jezebel Delias eucharis (Drury) 66-83 mm Common Dendrophthoe falcata
19 Psyche Leptosia nina (Fabricius) 35-50 mm Common Cleome viscosa
FAMILY NYMPHALIDAE
20 Blue Tiger Tirumala limniace (Cramer) 90-100 mm Common Lantana camara, Ageratum conyzoides,
Crotalaria retusa
21 Dark Blue Tiger Tirumala septentrionis (Butler) 75-95 mm Common Ageratum conyzoides
22 Stripped Tiger Danaus genutia (Cramer) 72-100 mm Common Tridax procumbens, Lantana spp.,
Crotalaria retusa
23 Plain Tiger Danaus chrysippus (Linnaeus) 70-80 mm Common Calotropis spp., Ageratum conyzoides,
Tridax procumbens,Crotalaria retusa,
24 Glassy Blue Tiger Parantica aglea (Stoll) 70-85 mm Common Calotropis spp., Ageratum conyzoides,
770
Stachytarpheta spp., Crotalaria retusa
25 Common Crow Euploea core (Cramer) 85-95 mm Common Ichnocarpus frutescens, Hemidesmus
771
indicus, Ficus spp., Streblus asper,
Ageratum conyzoides, Crotalaria spp.,
Chromolaena odorata
26 Common Nawab Polyura athamas (Drury) 60-75 mm Common Acacia pennata, Adenanthera pavonina
27 Common Evening Melanitis leda (Linnaeus) 60-80 mm Common Oryza sativa, Panicum spp.
Brown
28 Bamboo Treebrown Lethe europa (Fabricius) 65-75 mm Common Bambusa spp.
29 Common Palmfly Elymnias hypermnestra (Linnaeus) 60-80 mm Common Areca catechu, Cocos nucifera
30 Common Bushbrown Mycalesis perseus (Fabricius) 38-55 mm Common Oryza spp.
31 smooth-eyed bushbrown Orsotriaena medus (Fabricius) 45-55 mm Common Oryza sativa
32 Common Fivering Ypthima baldus (Fabricius) 32-48 mm Common
33 Common Fourring Ypthima huebneri (Kirby) 30-40 mm Common Grasses
34 Tawny Coster Acraea violae (Fabricius) 50-65 mm Common Aporosa lindleyana, Passiflora foetida
35 Tamil Yeoman Cirrochroa thais (Fabricius) 60-75 mm Common Hydnocarpus spp.
36 Rustic Cupha erymanthis (Drury) 50-60 mm Common
37 Common Leopard Phalanta phalantha (Drury) 50-60 mm Common
38 Commander Moduza procris (Cramer) 60-75 mm Common Ochreinauclea missionis, Mussaenda
frondosa
39 Common Lascar Pantoporia hordonia (Stoll) 45-50 mm Common Acacia pennata
40 Common Sailor Neptis hylas (Linnaeus) 50-60 mm Common Dalbergia spp., Zizyphus spp., Thespesia
populnea, Grewia spp., Bombax alabaricum
41 Clipper Parthenos sylvia (Cramer) 95-130 mm Common tinospora cordifolia
42 Common Baron Euthalia aconthea (Hewitson ) 55-80 mm Common Anacardium occidentalis, Mangifera indica,
Streblus asper
43 Grey Count Tanaecia lepidea (Butler) 65-85 mm Common Careya arborea
44 Common Map Cyrestis thyodamas (Boisduval) 50-60 mm Not Common Ficus spp.
45 Painted Lady Vanessa cardui (Linnaeus) 55-70 mm Common Blumea spp.
46 Angled Caster Ariadne ariadne (Linnaeus) 45-60mm Uncommon Ricinus communis
47 Common Caster Ariadne merione (Cramer) 45-60mm Common Ricinus communis
48 Chocolate Pansy Junonia iphita (Cramer) 55-80 mm Common
49 Grey Pansy Junonia atlites (Linnaeus) 55-65 mm Common
50 Peacock Pansy Junonia almana (Linnaeus) 60-65 mm Common Osbeckia spp.
51 Lemon Pansy Junonia lemonias (Linnaeus) 40-60 mm Common Sida rhombifolia
52 Great Eggfly Hypolimnas bolina (Linnaeus) 70-110 mm Common Sida rhombifolia, Hibiscus spp.
53 Danaid Eggfly* Hypolimnas misippus (Linnaeus) 70-85 mm Common Hibiscus spp.
FAMILY LYCAENIDAE
54 Ape Fly Spalgis epius (Westwood) 20-30 mm Not common Carnivorous caterpillars feed on mealy bugs
55 Indian Sunbeam Curetis thetis (Hbner) 40-48 mm Not rare Abrus precatorius, Pongamia pinnata
56 Red Spot Zesius chrysomallus (Hbner) 38-44 mm Not rare Caterpillars feed on ant larvae
57 Yamfly Loxura atymnus (Cramer) 36-40 mm Common Smilax spp., Dioscorea pentaphylla
Vincy et al., 2013

58 Monkey Puzzle Rathinda amor (Fabricius) 26-28 mm Not rare Ixora spp.
59 Slate Flash Rapala manea (Hewitson) 30-33 mm Common Acacia pennata
60 Common Silverline Cigaritis vulcanus (Fabricius) 26-34 mm Common Zizyphus rugosa, Canthium oromandelicum,
Clerodendrum inerme
61 Angled Pierrot Caleta caleta (Hewitson) 26-32 mm Not rare Zizyphus rugosa
62 Banded Blue Pierrot Discolampa ethion (Cramer) 26-30 mm Common Ziziphus spp.
63 Common Pierrot Castalius rosimon (Fabricius) 24-34 mm Common Zizyphus rugosa, Z. jujuba
64 Common Line Blue Prosotas nora (C. Felder) 18-25 mm Common Acacia catechu, Mimosa spp.
Vincy et al., 2013
65 Dark Cerulean Jamides bochus (Stoll) 25-34 mm Common Butea monosperma, Crotalaria spp.,
Pongamia pinnata
66 Common Cerulean Jamides celeno (Cramer) 27-40 mm Common Butea monosperma, Pongamia pinnata,
Abrus precatorius
67 Forget me not Catochrysops strabo (Fabricius) 25-35 mm Common Desmodium spp.
68 Lesser Grass Blue Zizina otis (Fabricius) 19-26 mm Common Fabaceae spp.
69 Red Pierrot Talicada nyseus (Gurin) 30-36 mm Common
70 Quaker Neopithecops zalmora (Butler) 16-30 mm Common Glycosmis pentaphylla
71 Plum Judy Abisara echerius (Stoll) 40-50 mm Common
FAMILY HESPERIIDAE
72 Common Spotted Flat Celaenorrhinus leucocera (Kollar) 45-55 mm Common

73 Indian Skipper Spialia galba (Fabricius) 20-27 mm Common Sida rhombifolia, Hibiscus spp.
74 Common Small Flat Sarangesa dasahara (Moore) 26-35 mm Common Asystasia spp.
75 Tricolour Pied Flat Coladenia indrani (Moore) 40-46 mm Common Mallotus philippinensis, Desmodium spp.
76 Suffused Snow Flat Tagiades gana (Moore) 45-50 mm Not rare
77 Water Snow Flat Tagiades litigiosa (Mschler) 37-44 mm Not rare Smilax spp.
78 Tamil Grass Dart Taractrocera ceramas 23-30 mm Common Oryza sativa, grasses
79 Common Grass Dart Taractrocera maevius (Fabricius) 22-28 mm Common Grasses
80 Common Dartlet Oriens goloides (Moore) 24-28 mm Common
81 Dark Palm Dart Telicota ancilla (Herrrich-Schffer) 33-36 mm Common Cocos nucifera, Oryza spp., Saccharum spp.
82 Rice Swift Borbo cinnara (Wallace) 30-36 mm Common Oryza sativa, Pennisetum spp., Ischaemum
spp., Cymbopogon spp.
83 Contigous Swift Polytremis lubricans (Herrrich- 36-42 mm Not common
Schffer)
84 Indian Palm Bob Suastus gremius (Fabricius) 32-45 mm Common Calamus spp., Caryota urens, Cocos nucifera
85 Gaint Red eye Gangara thyrsis Fabricius) 70-76 mm Not rare Calamus rotang, Caryota urens, Cocos nucifera
86 Common Redeye Matapa aria (Moore) 40-55 mm Common
87 Chestnut Bob Iambrix salsala (Moore) 26-30 mm Common Grasses and bamboos
88 Restricted Demon Notocrypta curvifascia (Felder & 38-50 mm Common Costus speciosus
Felder)
89 Grass Demon Udaspus folus (Cramer) 40-48 mm Common Zingiber spp.
90 Pygmy Scrub Hopper** Aeromachus pygmaeus (Fabricius) 20-22 mm Common
91 Indian Ace Halpe homolea (Hewitson) 30-36 mm Common Bamboo
772
* - indicates species coming under Schedule I Part IV and ** - Schedule II Part II of The Wildlife (Protection) Act, 1972
Vincy et al., 2013
The study shows that the sustained interference Mathew G and Rahamathulla VK. 1993. Studies on
and disturbance seem to affect the occurrence and the butterflies of Silent Valley National Park. Entomon,
numerical strength of each butterfly species. If this 18:185-192.
situation goes unabated, the abundant butterflies may
Larsen TB. 1987a. The butterflies of the Nilgiri
become rare and the less abundant ones could disappear
mountains of South India (Lepidoptera: Rhopalocera).
permanently. Further, the decline in the number of
Journal of the Bombay Natural History Society, 84: 26-
butterflies largely allows inbreeding which becomes fatal
43.
in course of time. Modified habitats with reduced plant
cover contribute to warm conditions and these conditions Larsen TB. 1987b. The butterflies of the Nilgiri
might allow some butterflies to extend their distribution mountains of South India (Lepidoptera: Rhopalocera).
to different habitats. The butterflies which control certain Journal of the Bombay Natural History Society, 84: 291-
plant pets, if decline in number or disappear from the 316.
habitat, plants too get affected because of the unchecked
Larsen TB. 1987c. The butterflies of the Nilgiri
plant pets. Therefore, the very presence of butterflies in
mountains of South India (Lepidoptera: Rhopalocera).
species and number may be taken as an indication of the
Journal of the Bombay Natural History Society, 84: 560-
health of the habitat.
584.
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Arora K. 2003. Forest Laws. The Wildlife Protection mountains of South India (Lepidoptera: Rhopalocera).
Act, 1972 as amended by the Wildlife (Protection) Journal of the Bombay Natural History Society, 85: 26-
Amendment Act, 2002 (Act 16 of 2003). Published by 43.
Professional Book Publishers, New Delhi, 85.
Lawton, JH Bignell D E Bolton B Bloemers GF
Chakravarthy AK Rajagopal D and Jagannatha R. Eggleton P Hammond PM Hodda M Holts RD
1997. Insects as bio indicators of conservation in the Larsen TB Mawdsley NA Stork NE Srivastava DS
tropics. Zoos Print, 12:21-25. and Watt AD. 1998. Biodiversity inventories indicator
taxa and effect of habitat modification in tropical forest.
Evans WH. 1932. Identification of Indian Butterflies.
Nature, 391: 72-76.
Bombay Natural History Society, Bombay, 454.
Palot J Balakrishnan VC and Kambrath B. 2003.
Gay T Kehimkar ID and Punetha JC. 1992. Common
Keralathile Chitrasalabhangal. Malabar Natural History
Butterflies of India. Oxford University Press, Bombay.
Society, Calicut, Kerala, 195.
Gunathilagaraj K. 1998. Some South Indian Butterflies.
Sudheendrakumar VV Binoy CF Suresh PV and
Tamil Nadu, India: Nilgiri Wildlife and Environments
Mathew G. 2000. Habitat associations of butterflies in
Association, Udhagamandalam, Nilgiris, 274.
the Parambikulam Wildlife Sanctuary, Kerala, India.
Haribal M. 1992. The butterflies of Sikkim, Himalaya Journal of the Bombay Natural History Society, 97: 193-
and their natural history. Nataraj Publishers, Dehradun, 201.
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Wynter-Blyth MA. 1957. Butterflies of Indian Region.

Bombay Natural History Society, Bombay.
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OVER VIEW
Microbial production of glutaminase enzyme

Authors: ABSTRACT:
Mario Khalil Habeeb
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
Institution: characteristics over other sources due to their easy production on a commercial scale
Microbiology Department,
and stability. Different microorganisms are known to produce various enzymes such as
Faculty of Science, Ain
Shams University, 15566 bacteria, fungi and actinomycetes which produce a variety of extra-cellular and endo-
El-Khalifa El-Mamoun cellular enzymes. Some of these actinomycetes enzymes have been isolated from the
street, Abbassia, Cairo, culture filtrates or the mycelium, concentrated and purified. Others have only been
Egypt, Postal code: 11566. 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
Corresponding author: enhancer in food industry by increasing the amount of glutamic acid content in the
Mario Khalil Habeeb. fermented food .
Email: Keywords:
mario_khalil87@yahoo.com, Actinomycetes, Anticancer properties, Enzymes, Glutamic acid and
mario_khalil@sci.asu.edu.eg L-Glutaminase.
Telephone: Article Citation:

+20 (02) 22409635 Mario Khalil Habeeb.
Microbial production of glutaminase enzyme.
Mobile:
+20 (0128) 3941815 Dates:
Received: 16 Jan 2013 Accepted: 22 Jan 2013 Published: 06 Feb 2013
Web Address:
http://jresearchbiology.com/
documents/RA0325.pdf. This Open Access article is governed by the Creative Commons Attribution License (http://
cited.
775-779 | JRB | 2013 | Vol 3 | No 1

Habeeb., 2013
INTRODUCTION starvation of cancerous cells and their possible death

Enzymes are highly selective catalytic proteins (Santana et al., 1968).
produced by living cells which may or may not contain a Glutaminase Producing microorganisms
non-protein prosthetic group (Underkofler et al., 1958). Different types of organisms were reported
Actinomycetes are considered to be preferred to produce glutaminase enzyme. However, The selection
enzymes sources due to their production of extracellular of the right organism is very critical to obtain high yield
enzymes. They are highly diverse group with numerous of the required enzyme (Akujobi et al., 2012).
members representing important source of microbial L-glutaminases from E. coli, Pseudomonas sp.,
enzymes. Actinomycetes genera are differentiated from Bacillus sp., and Clostridium welchii have been isolated
each other based on morphological, biochemical, and and well studied (Wade et al., 1971). In addition to these
physiological criteria. They act as decomposers of bacterial sources, the fungus Aspergillus oryzae showed
complex animal and plant materials resulting in release a great ability to produce this enzyme. Among
of simple substances, especially carbon and nitrogen microorganisms, actinomycetes are widely recognized
which is easily utilized by other organisms, thus as preferable L-glutaminase sources because they
performing a vital role in life cycle. Due to their generally produce extracellular enzymes, which facilitate
significant biochemical activities, Actinomycetes are the enzyme recovery from the fermentation broth
used in commercial production of various substances such as glutaminase from Streptomyces rimosus
such as antibiotics and enzymes (Waksman, 1950). (Sivakumar et al., 2006).
Because of its industrial and pharmaceutical Microbial Glutaminase Characteristics
applications, intensive research was conducted on Temperature is considered to be an important
L-glutaminase recently. L-glutaminase is produced by factor affecting the enzyme stability, The optimum
various terrestrial microorganisms such as Pseudomonas temperature recorded by many glutaminases ranged from
sp., Acinetobacter sp., Escherichia coli, Bacillus sp., 40-50C. However, the temperature stability of
Hansenula sp., Candida sp., Aspergillus oryzae and glutaminase I (Micrococcus glutaminase) of M. luteus
Beauveria bassiana (Sabu, 2003). Also few could be increased by the addition of 10% NaCl
marine microorganisms such as Micrococcus luteus, (Moriguchi et al., 1994). The optimum temperature for
Vibrio cholera and Pseudomonas fluorescens were A. oryzae glutaminase was around 37-45C and remained
reported to produce the enzyme (Chandrasekaran, 1997). stable at up to 45C and the enzyme was completely
Definition inactive at 55C (Nakadai and Nasuno, 1989).
L-glutaminase is classified as an amidohydrolase It is interesting that the exposure of E. coli
enzyme which acts upon amide bonds of L-glutamine glutaminase B to cold resulted in a reversible
generating L-glutamic acid and ammonia. It is present inactivation of enzymatic activity, while subsequent
in both microorganisms and mammalian tissues warming to 24C restored the activity. There was no
(Ohshima et al., 1976b). Microbial sources of difference in the molecular weight of the cold inactivated
glutaminase showed a great role in various applications enzyme and the warm activated enzyme. The
such as its use in fermented foods precisely in soy sauce conformational changes which probably occur upon
and other related types, in addition to its use as exposure to cold resulted from a weakening of the
anticancer agent which act by inhibition of glutamine interaction among hydrophobic groups in the protein
utilization by the cancerous cells resulting in selective (Chou et al., 1993).
Habeeb., 2013
The salt-tolerance of glutaminase is an important By using this method it was found that L-glutaminase
parameter in industrial processes that include producing marine alkalophilic Streptomyces sp. SBU1
high-salinity. It was reported that the high-salt which was isolated from Cape Comorin coast,
concentration (nearly 3 M NaCl) used in the process India gave highest enzyme production after 4 days
of soy sauce fermentation resulted in remarkable of incubation and at 14% Corn steep liquor
inhibition of the koji mold (A. oryzae) Glutaminase (Krishnakumar et al., 2011).
(Koibuchi et al., 2000). Applications
Methods Used for Microbial Glutaminase Production L-glutaminase has received great attention due to
Two methods are known for the production of its valuable applications in several fields especially in
microbial glutaminase. medicine and its use as an anticancer agent either alone
Submerged (Liquid) Production Method or together with any other agents is known as enzyme
In this method, the sterile media together with therapy, In addition to its role as flavor enhancer by
the enzyme producing organism were introduced into increasing the glutamic acid content of food. Also
large fermentors (Tanks) followed by constant mixing glutaminase applications extend to the enzyme utilization
and supply of sterile air (Schuegerl et al., 1991). as biosensor in analytical purposes by measuring the
Actinomycetes glutaminases showed a high salt levels of L-glutamine and finally in the manufacture of
tolerance in this production method. Reports showed that fine chemicals such as theanine when used with bakers
Streptomyces rimosus isolated from estuarine fish yeast.
recorded high salt tolerance and the highest enzyme Glutaminase as Enzyme Therapy
production obtained at temperature 27C, pH 9.0 and Glutaminase can be used as alternative for cancer
both glucose and malt extract proved to be the best treatment as enzyme therapy. The mechanism for
carbon and nitrogen sources for maximum enzyme glutaminase therapy includes that L-Glutaminase act on
production (Imada et al., 1973). its substrate (L-glutamine) and breaks it down leading to
Surface Production Method the selective destruction of the tumor cells accompanied
This method includes the use of solid support on by inhibition of both protein and nucleic acid
which microorganisms are grown. Surface production biosynthesis due to glutamine starvation and this is
method (solid state fermentation) showed 25 to 30 fold attributed to the inability of cancerous cells to synthesis
increase in enzyme production when compared with glutamine (Tanaka et al., 1988). This is due to the fact
submerged production (Sabu et al., 2000b). that some types of cancerous cells utilize glutamine
Wheat bran was found to be a favorable support greatly (Lazarus and Panasci, 1986). Concerning this
for microorganisms in the process of glutaminase finding various enzymatic therapies developed to deprive
production (Kashyap et al., 2002). In addition to wheat L-glutamine to cancerous cells (Roberts et al., 1970).
bran many other solid supports showed high efficiency in Glutaminase as Flavor Enhancer
the enzyme production such as ground nut cake powder, Glutamate is a famous amino acid and
copra cake powder and sesamum oil cake (Prabhu and considered as a natural constituent of many fermented or
Chandrasekaran, 1995). Polystyrene beads, supported by aged foods, such as soy sauce, fermented bean paste and
mineral salts and glutamine are another form of solid cheese (O`Mahony and Ishi, 1987). It gives these types
supports used for the enzyme production. 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 REFERENCES

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Ohshima M, Yamamoto T and Soda K. 1976b. Tanaka S, Robinson EA, Appella E, Miller M,
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Original Research
A review on the role of nutrients in development and

organization of periphyton
Authors: ABSTRACT:
Saikia SK1 , Nandi S2 ,
Majumder S3 .
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
Institution:
1. Assistant Professor, and also outlines its ecological role in nutrient sharing with other aquatic components.
Aquatic Ecology Laboratory, The development and succession of periphyton is a function of nutrient and carbon (C)
Department of Zoology, sharing with its constituent parts and ambient environment. Through mechanisms like
Visva Bharati University, entrapment, de novo synthesis, nutrient leakage, trophic upgrading etc., ambient
Santiniketan, West Bengal, nutrients are routed to periphyton and transferred to upper trophic levels. Periphyton
India, Pin-731235. communities stand next to phytoplankton for their contribution to primary
productivity, in nutrient rich aquatic environment. Unlike phytoplankton, nutrient
2. Research Fellows,
poor aquatic environment has no effect on periphytic primary productivity.
Aquatic Ecology Laboratory,
Department of Zoology, As periphyton communities are attached to substratum, their ability to assimilate
Visva Bharati University, organic nutrient through substratum is an additional advantage over phytoplankton.
Santiniketan, West Bengal,
India.
3. Research Fellows,
Aquatic Ecology Laboratory,
Department of Zoology,
Keywords:
Visva Bharati University,
Santiniketan, West Bengal. Aquatic ecosystem, Biofilm, carbon, primary productivity, phytoplankton.
Corresponding author: Article Citation:

Saikia SK. Saikia SK, Nandi S, Majumder S.
A review on the role of nutrients in development and organization of periphyton.
Email: Dates:
surjyasurjya@gmail.com Received: 20 Nov 2012 Accepted: 10 Dec 2012 Published: 11 Feb 2013
Web Address:
http://jresearchbiology.com/ This Open Access article is governed by the Creative Commons Attribution License (http://
documents/RA0307.pdf. commercial, distribution and reproduction in all medium, provided the original work is properly
cited.
780-788 | JRB | 2013 | Vol 3 | No 1

Saikia et al., 2013
INTRODUCTION contributor of most of the nutrient inputs to aquatic

The term periphyton (peri round; phyton plant) ecological cycles. The present review is an attempt to
was proposed by (Behning, 1924) and popularized outline periphyton as an integral and essential component
by several authors (Cooke, 1956; Sladeckova, 1962; of aquatic ecosystem highlighting few areas recently
Pieczynska, 1970). There exist a series of definitions addressed on the role of periphyton in nutrient sharing in
proposed for periphyton (Young, 1945; Neel, 1953; aquatic processes.
Wetzel, 1963). (Wetzel, 1983) defined it as the micro Periphyton: A Nutrient Dependent Organization
floral community living attached to any substrate (Whal, 1989) discussed settling pattern
under water. (Stevenson, 1996) used it for describing of biofilm (Figure 1), in four phases: (i) surface
microorganisms such as algae and bacteria growing in conditioning or adsorption of dissolved organic
association with substrata. These communities play an compounds where macromolecules attach to submerged
important role in water bodies, not only as important surfaces following a spontaneous physical-chemical
primary producers and energy source for higher trophic process; (ii) primary colonization or bacterial settling
levels, but also by affecting the nutrient turnover and the following surface conditioning and after their
transfer of nutrients between the benthic and the pelagic colonization, bacteria start to produce EPS,
zone (review Saikia, 2011). The substrates of periphyton (iii) secondary colonization to bacterial layer and EPS
commonly include submerged plants or plant parts, pool by eukaryotic unicellular microorganisms, mainly
rocks and sediments. Such substrate selection designs protozoan, microalgae and cyanobacteria and (iv) settling
periphyton as a medium in transferring and trophic of eukaryotic multicellular organisms as a function of
upgrading of nutrients. This property recognizes nutrient sharing, grazing and predation. According to
periphyton as a tool for biofiltering excess nutrients from (Wetzel, 1983), associated organisation from secondary
polluted waters and for efficient nutrient transfer in colonization onwards can be designated as periphyton.
aquatic food chain. In that way, it could be defined as an advanced
Aquatic ecosystems mainly comprise of successional stage of biofilm. However, there could be
freshwater and marine water bodies and their ecology a fifth (v) phase; the tertiary colonization where
discusses the relationship of aquatic organisms and bacterioplankton colonized on the surfaces of unicellular
its interaction with the immediate environment. The and filamentous secondary colonizer (e.g. diatom,
principle biotic components primarily explained in the Oedogonium etc.). Several bacteria different from early
recent past for their contributions to different interactions colonizer settle on algal surfaces at this stage
in aquatic ecosystem are macrophytes, plankton (Alldredge et al., 1993; Armstrong et al., 2000).
(Zooplankton and phytoplankton) and invertebrates Periphyton are rich Carbon source
(benthos, nekton and neuston). Till the mid of TEP with rich Carbon sources of
th
19 century, periphyton or associated organisms were Glucopolysaccharides in aquatic environments
not given any biological credit for their role in aquatic initiates early colonization of bacteria through
ecosystem. Probably, (Wetzel, 1963) in his evolutionary surface conditioning (Stoderegger and Herndl, 1999).
review paper Primary productivity of periphyton in The bacterial EPS from early biofilm exists as a part of
Nature, for the first time made convincing remark on the dissolved organic matter (Lignell, 1990) as well as
role of periphyton in aquatic ecosystem. Even today, particulate matter (Decho, 2000). It acts both as rich
periphytic communities are ignored as a major organic Carbon storage (Freeman and Lock, 1995) and
Saikia et al., 2013
chief supplier of Carbon demand for organisms that feed management of community metabolism of periphytic
on periphytic aggregates (Decho and Moriarty, 1990; matrix and can trap the metabolic products released
Hoskins et al., 2003). Being polyanionic in nature by bacterta on algal surface (Makk et al., 2003). Such
(Costerton et al., 1978), EPS further permits inorganic algae-bacteria interactions enrich periphytic organic
nutrient entrapment through ion exchange processes matrix with components of polysaccharides, proteins,
(Freeman et al., 1995) leading to storage of organic nucleic acid and other polymers (Davey and Otoole,
Carbon in the biofilm. In addition, among the bacterial 2000).
fractions, cyanobacteria are important primary producers Periphytic pathway of nutrient transfer
and many of their species can x atmospheric Nitrogen The periphytic nutrient transfer pathway (PNP)
(Whitton and Potts, 1982). Chemical screening of mainly involves ambient nutrient entrapment, storage
laboratory grown, commercially viable cyanobacteria and transferring it to immediate higher trophic level. The
have revealed that they have a high nutritional value, in fate of PNP gets its initiation from the surface
terms of protein (Choi and Markakis, 1981). conditioning phase of periphyton formation. As soon as
During tertiary phase of periphyton development, TEP prepares the substrate surface for colonization,
algal communities play indirect role in nutrient addition bacteria as initial colonizer develops micro-colonies
to periphytic complex through their surfaces. A study on (Costerton, 1984) and through EPS, it supplies a
algae-bacteria interactions on biotic surfaces revealed significant source of Carbon to periphytic complex
that bacterial abundance is significantly higher in areas (Hobbie and Lee, 1980) (Figure 2). A PNP establishes
of diatom colonization on substrates (Donnelly and between dissolved organic in periphytic complex and
Herbert, 1999). These bacteria contribute to the 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
Figure 1. Formation of periphytic complex on from tertiary phase of colonization. The second
natural substrate showing tertiary phase of coloni-
mechanism consists of endogenous energy reserves.
zation (Modified from Whale, 1989). TEP, Trans-
parent Exopolymer Prticles; EPS, Extracellular These reserves consist of Carbon that is accumulated and
Polymeric Substances
assimilated inside the microbial cell and can be

Saikia et al., 2013
mobilized to ensure survival during starvation (Dawes physiological processes in living organisms and are
and Senior, 1973) and thereby recovery of periphytic major nutrient constituents of polar lipids, and are
aggregates due to senescence. The third mechanism of present in cell and chloroplast membranes. The
organic Carbon storage is the polysaccharide exudates dominance of algae in periphytic canopy acts as rich
(Freeman and Lock, 1995) released by algae at tertiary source of FA to animals grazing on periphyton.
phase under nutrient (especially Phosphorus) limited Primary productivity of periphyton
condition. The algal components release polysaccharide The energetic relation of an ecosystem is
exudates to EPS under Phosphorus limitation on which principally regulated by primary production. In aquatic
tertiary phase bacteria flourish. In return, these bacteria ecosystem, algae are dominant primary producers, and
remineralize Phosphorus for algae. In addition, the ECM responsible for both Carbon fixation and sequestration.
with polyanionic by nature (Costerton et al., 1978) is Periphyton with majority of algae might have significant
believed to permit nutrient entrapment through ion contribution to primary production of aquatic ecosystem.
exchange processes (Freeman et al., 1995). (Freeman However, very few investigations have been performed
and Lock, 1995) proposed that the entrapment on measurements of photosynthetic rates of algal
mechanism may also permit the storage of organic periphyton under natural conditions. (Wetzel, 1963)
Carbon in the biofilm. pointed out technical/methodological difficulty in
In transferring nutrient through PNP, the assessing such parameters of periphyton under natural
bacterial Carbon enters to organisms in the next trophic condition. From an analysis on nutrient limiting and
level as complex Carbon rich compounds. The Fatty acid nutrient rich lakes, it is obvious that periphyton
(FA) component of algae is under extensive research productivity contributes more than 30% of primary
now a day as Carbon rich compounds. Periphytic matrix productivity to the aquatic ecosystem (Figure 2a). On
is dominated by algae and hence FA contributes to the comparison, it seems evident that the nutrient limited
food quality of matured periphytic organization. In algae, aquatic ecosystems have more or less equal primary
FA increases as a result of exposure to stressful productivity levels to nutrient rich aquatic ecosystems.
environmental conditions, such as high temperature, The same is not true in case of phytoplankton
nutrient extremes and harsh light conditions. (Figure 2b).
Polyunsaturated fatty acids (PUFAs) also affect many
Annual Primary Production in
percentage
Phytoplankton Periphyton Macrophyte
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).

Saikia et al., 2013
Nutrient Regulated Biotic Interactions of Periphyton limited environments, it relies mainly on organic
Biotic interactions in aquatic ecosystems are nutrients from natural substrate. All artificial substrates
more complex than any other ecosystems for its variable cannot serve as organic nutrient supplier to periphyton.
nature. Interactions between periphyton and biotic Substrates like sediments or seed grains acts as nutrient
components in aquatic ecosystem are primarily regulated diffusing substrate releasing nutrients to overlying
by nutrients and can be discussed under following periphytic layer. (Hansson, 1989) showed that epipelion
subheadings- can significantly lower nutrient availability in the water
Plankton-periphyton interaction column due to uptake of diffusing nutrients. (Hagerthey
Periphyton-macrophyte interaction and Kerfoot, 1998) demonstrated that inflowing ground
Grazer-periphyton interaction water is a significant source of nutrients for episammon
Plankton-Periphyton interaction in nutrient limiting environment. These sediments act as
The plankton-periphyton interaction is better nutrient source for periphyton (Burkholder, 1996).
principally regulated by light and nutrient availability in Substrate based nutrient uptake by periphyton is further
the environment. Both the communities are composed of related to depth, light availability, physical disturbances
common members of bacterial, algal and zooplanktonic etc.
origin. However, on spatial ground, habitats of both Grazer-Periphyton interaction
plankton and periphyton have differences in receiving Studies reported that several herbivore types
light and nutrients. Conceptual models revealed that (e.g. gastropods, trichopteran larvae and fish) can
nutrient limited environments are dominated by dramatically reduce periphytic biomass to only a few
periphyton than plankton (Wetzel, 2001; Hansson, 1992). percent of total biomass (Hillebrand et al., 2000).
Nutrient limitation results thin planktonic cover that Although grazing results reduction in periphytic biomass,
allows maximum light to pass through water column to the total productivity of the periphytic complex increases
reach the bottom of the ecosystem facilitating due to reduced competition among algal members
multiplication of periphytic population. Conversely, (Carpenter, 1986; Mc Cormick and Stevenson, 1989).
plankton rich aquatic ecosystems limit growth of (Norberg 1999), using transparent incubation chambers,
periphyton due to limited light availability. Epiphytic measured a 4-fold increase in periphyton specific
communities can better adsorb nutrients from sediments productivity in grazed periphyton compared to ungrazed
or bottom of the system through macrophytes controls. Moreover, the grazer presence increased the
(Burkholder and Wetzel, 1990). Chlorophyll: biovolume ratio, especially reported from
Periphyton-substrate interaction streams (Hill and Knight, 1987). In addition to increase
Substrate type plays a driving role in growth and in productivity, grazing and competition can modify the
succession of periphyton. Being a substrate based species composition of periphytic algal assemblages
organization, periphyton have access to both organic (Duffy and Hay, 2000; Nielsen, 2001), generating
nutrients from substrate and inorganic nutrients from heterogeneity through temporal or spatial scale on the
water column. In nutrient rich environments, it receives substrate. A top down effect of consumers on their prey
nutrients from water column (Eminson and Moss, 1980; can be further accelerated by grazer and grazer excretion
Burkholder, 1996). Here, similar to planktonic cells, of nutrients, removal of senescent cells, or increased
periphytic cells can use inorganic nutrients efficiently, uptake of nutrients by the remaining cells (Lamberti
specifically dissolved organic Phosphorus and in nutrient et al., 1987; Kahlert and Baunsgaard, 1999). Grazers

Saikia et al., 2013
may have strongest effects on Carbon:Phosphorus CONCLUSION

and Nitrogen:Phosphorus, but Carbon:Nitrogen and Disrupted nutrient cycling is a major problem
Carbon:Chlorophyll may remain unaffected (Hillebrand both in freshwater and marine ecosystems and
and Kahlert, 2001). Hillebrand et al., (2008) described periphyton could be a non-point manager of nutrient
three pathways for grazer mediated periphytic cycle disruption and hence can overplay on plankton for
interactions affecting nutrient stoichiometry. First, the nutrient cycling in aquatic ecosystem. During renovative
non algal component, which could be a dominant part practices, strategies of aquatic ecosystem health
of the organic material of periphyton assemblage managers greatly ignore the role of these substrate based
(Frost et al., 2002) is reduced by unselective grazing. microorganisms. At the same time, it can play as an
Benthic invertebrates graze upon both detritus and algal efficient supplier of nutrient to its grazer under
component of periphyton but only algae regenerate. controlled and well managed productive practices. It is
Therefore, grazing not only reduces non algal component observed that at traditional level, farmers from different
of periphyton, but also facilitates the growth of live parts of the world have been practicing periphyton to
component within it. (Jones et al., 1999) suggested that feed aquacrops to convert periphytic energy biomass to
epiphytes can influence the nutritional quality of the crop biomass (Saikia and Das, 2009). Such conversion of
periphyton which grows on their surfaces, making it biomass is an outcome of increased assimilation of
more nutritious for grazing by invertebrates, particularly micro- and macro nutrients from periphytic complex in
snails. In return, these grazers might preferentially feed the fish body through trophic upgrading (Saikia and
on the periphyton and clear the plants of a potential Nandi, 2010). Further researches on the mode of energy
competitor for nutrients, with the plants and grazers both transfer through periphytic food chain, enhanced nutrient
gaining from this relationship. Secondly, in streams, uptake under manipulative nutrient input, modelling on
nutrient uptake of intact periphyton mats is often slower applied periphytic ecology, ecotoxicology, Carbon
than cell specific uptake rates as boundary effects reduce entrapment and delivery, directing nutrient and Carbon
the uptake ability of the benthic algae (Riber and Wetzel, sequestration both in marine and freshwater are needed
1987; Bothwell, 1989; Burkholder et al., 1990). Grazer for better understanding of its role in aquatic ecosystem.
presence alters periphyton architecture, increases
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Journal of Research in Biology An International Scientific Research Journal
Original Research
Assessing heavy metal contamination of road side soil in urban area

Authors: ABSTRACT:
Sarala Thambavani D1,
Vidya Vathana M2. 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
Institution: road side environment. The present research is conducted to study heavy metal
1. Associate Professor contamination in road side and industrial soil of Madurai city. The soil samples are
Department of Chemistry, collected from three sites and analyzed for six heavy metals (Pb, Cu, Cr, Zn, Ni and Cd).
Sri Meenakshi Govt. Arts Their concentration and distribution in different depths (0 cm, 5 cm and 10 cm) were
College (W), Madurai. determined. Heavy metal contents were analyzed by Atomic Absorption Spectroscopy
(AAS). The studies with Enrichment Factor (EF) indicate that lead has been enriched to
2. Assistant Professor,
quite great extent while the Normalized Scatter Coefficient values (NSC) indicate
Department of Chemistry,
Sacs M.A.V.M.M Engg. faster enrichment of cadmium. The level of heavy metals in road side soils were higher
College, Madurai. 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.

Sarala Thambavani D. Pollution, combustion, heavy metal enrichment, road side soils, enrichment
factor, Normalized scatter coefficient value, environmental pollution.
Web Address: Article Citation:

http://jresearchbiology.com/ Sarala Thambavani D and Vidya Vathana M.
documents/RA0187.pdf. Assessing heavy metal contamination of road side soil in urban area.
Dates:
Received: 16 Jan 2012 Accepted: 27 Jan 2012 Published: 16 Feb 2013
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.
789-796 | JRB | 2013 | Vol 3 | No 1

An International Scientific
Thambavani and Vathana, 2013
INTRODUCTION It is needless to say that the industrial activities

Pollution in recent years has increased in the metropolitan cities of the world are responsible for
considerably as a result of increasing human activities the addition of pollutants through chemical factories,
such as burning of fossil fuels, industrial and automobile residential activities (Point sources) and vehicular traffic
exhaust emissions. The pollution of soils by (non-point sources) which are the primary sources of soil
heavy metals from automobile sources is a serious pollution. The objective of this study, is to investigate the
environmental issue. The majority of the heavy metals effect of heavy metal pollution of soil along road sides.
are toxic to the living organisms and even those The present study reports the role of industrial and
considered as essential can be toxic if present in excess. urban activities in the heavy metal contamination of the
The heavy metals can impair important biochemical soils in the Madurai industrial area with the objectives:
processes posing a threat to human health, plant growth To assess the extent of heavy metal pollution
and animal life (Jarup 2003; Michalke 2003; Silva et al., influenced by urban and industrial activities.
2005). To predict the rate of heavy metals pollution in the
The waste products from vehicles that ply future if the activities are allowed with the same
highways contain some heavy metals inform of smokes. pace.
Emissions from exhaust pipes of automobile engine and To understand the variations in the behavior of
contacts between different metallic objects in machines different heavy metal.
contain such heavy metals as Lead (Pb), Zinc(Zn), Iron
(Fe), Copper (Cu), Chromium (Cr) and Cadmium (Cd) METHODS
and are major sources of pollution among soils (Turer Field Methodolgy
and Maynard, 2003). To understand the state of environment of the
Soils are usually regarded as an ultimate sink. Madurai area a detailed field survey was carried out and
For heavy metals discharged into the environment (Banat after having identified possible sources of pollution a
et al., 2005) and sediments can be sensitive indicators for part of Madurai area was selected. This area is under
monitoring contaminants in aquatic environment (Pekey intense human interference in terms of growing
et al., 2004). Therefore the environmental problem of urbanization (municipal sewage sludge, traffic pollution
soil and sediment pollution by heavy metals has received in particular) and industrialization.
increasing attention in the last few decades in both Selection of sampling site
developing and developed countries throughout the In the present study stratified regular sampling
world (Zhang et al., 2007). Hence, inorder to monitor method was adopted for soil sample collection as in
heavy metal pollution in an area, due to the geo-assessment of the variables estimated, the stratified
anthropogenic activity (Sarala Thambavani and Vathana, regular sampling is more suitable because this kind of
2011), the soil samples represent an excellent media sampling draws homogenous error (Burgess et al., 1981).
because heavy metals are usually deposited in the top Different sampling stations were selected and samples
soil (Govil et al., 2001; Romic and Romic, 2003) and are collected from the top layer of the soil using plastic
help in knowing the sources of heavy metals and also spatula after removing the debris, rock pieces and
controlling and optimizing their effects on the human physical contaminants. In order to have the background
health. concentration values of the heavy metal elements, three
soil samples were collected, each from 100 cm below
ground level, which are least affected by anthropogenic RESULTS

activities (Table1). The samples were placed in the clean The concentration of heavy metals Lead,
polythene bags, which were brought to the laboratory. Copper, Chromium, Nickel and Cadmium in the soils
Laboratory Methodology of Madurai industrial, traffic and residential area
The samples were brought to the laboratory were analyzed, collected at six sampling stations during
where they are dried and mixed thoroughly to obtain the May 2011- Oct 2011. The range of the concentrations
representative samples. Soon after drying the debris and found in different sampling stations are (i) Pb industrial
other objects were hand picked up and the sample were (24.81-42.37 mg/kg), traffic (26.80-5.32 mg/kg)
grounds in a mortar to break up the aggregates or lumps, and residential (20.42-2.66 mg/kg) (ii) Cu industrial
taking care not to break actual soil particles. Soil samples (10.40-16.24 mg/kg), traffic (10.69-18.20 mg/kg)
were then passed through a 2 mm sieve in order to and residential (10.5 -18.16 mg/kg) (iii) Cr industrial
collect granulometric fraction. Since trace metals are (17.0-34.50 mg/kg), traffic (14.56-21.60 mg/kg)
often found mainly in clay and silt fractions of soil and and residential (25.12 mg/kg) (13.60-18.52 mg/kg)
hence the size fraction <63 m sieve (wet sieving) and ( i v) Zn in dustr ial (22.5-45.6 mg/kg),
was used to measure the concentration of the heavy traffic (22.32-25.46 mg/kg) and residential (22.24- 25.12
metals Lead, Copper, Chromium, Zinc, Nickel and mg/kg) (v) Ni industrial (11.85-14.0 mg/kg), traffic
Cadmium from all the samples collected. ( 11.52 -14.80 mg/kg) and residential (11.70-13.9 mg/kg)
For this purpose the clay and silt fraction were (vi) Cd industrial (1.24-4.32 mg/kg), traffic
digested by acids to get the solution by taking 5 g of (1.60-3.62 mg/kg) and residential (1.70-2.25 mg/kg).
sample into a 300 ml polypropylene wide-mouthed jar The mean concentration for these heavy metals
and distilled water was added to make a total 200 ml. from the surface soil have been calculated to be (i) Pb
Then it was acidified with 10 ml HF, 5 ml HClO4, industrial (33.23), traffic (41.50) and residential (24.31).
2.5 ml HCl and 2.5 ml HNO3 in order to completely (ii) Cu industrial (12.97), traffic (15.03) and residential
digest the soil. This jar was shaken on an orbital shaker (14.98). (iii) Cr industrial (24.33), traffic (17.53) and
for 16 h at 200-220 rpm before being filtered through residential (15.51). (iv) Zn industrial (29.78), traffic
whatman filter paper (No.42) into acid washed bottles. (24.23) and residential (23.74). (v) Ni industrial (12.77),
The solution was stored and heavy metal contents were traffic (13.72) and residential (12.99). (vi) Cd industrial
analyzed by Atomic Absorption Spectrophotometer as (2.94), traffic (2.59) and residential (1.92) respectively at
per the method recommended by committee of soil the confidence limits of 95%.
standard methods for analyses and measurement (1986). The concentration of heavy metals in all the
The raw data obtained during the course of laboratory sampling stations exhibit an increasing trend over a very
analyses were stored in Microsoft Excel software and short period of monitoring from May 2011-Oct 2011
further processed to obtain various parameters required (Figure 1). It was observed that the mean concentration
for interpretation. 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 DISCUSSION

and Cadmium. In order to evaluate the rate of accumulation of
Accumulative Signature of Heavy Metals heavy metals in the soils the mean values for all heavy
An increasing trend has been found for the heavy metals studied were considered along with Enrichment
metal elements Lead, Copper, Chromium, Zinc Nickel factor values of all six metals (Table 2), which clearly
and Cadmium wherein the Lead and Cadmium are indicate the highest enrichment of Cadmium followed by
getting accumulated with very rapid rate mainly due to Lead, Zinc, Chromium, Copper and Nickel in all the
anthropogenic activities (Sayadi, 2009). In order to three sampling stations of industrial, traffic and
assess the variations in the heavy metal accumulations in residential area. The values of NSC for all six heavy
the soils, the calculated measures that is Enrichment metal showed that Cadmium is increasing in soil
Factor and Normalized Scatter Coefficient were used. environment of industrial area followed by Zinc,
The Enrichment Factor (EF) is a ratio of the Chromium, Lead, Copper and Nickel. In traffic area
concentrations of the heavy metals in the soil samples to Lead is increasing in soil environment followed by
the corresponding concentration of natural background Cadmium, Chromium, Copper, Nickel and Zinc and in
concentration. EF is calculated with the help of the residential area Copper is increasing in soil environment
formula given by Subramanian and Datta dilip (1998) followed by Chromium, Cadmium, Lead, Nickel and
and presented in Table 2. Zinc.
EF = Value of a given metal concentration found on soil (mg/kg) It is observed that in all the sampling sites, Lead
Natural local background concentration of the metal (mg/kg) shows highest concentration in soil and also have high
Table 2 Enrichment Factor for heavy metals in the
Normalized Scatter Coefficient (NSC) has been soils
calculated to asses the temporal variability of the heavy
Industrial Area
metals in the soils. It helps to understand the increasing Pb Cu Cr Zn Ni Cd
or decreasing concentration of heavy metals in the soils 4.6 1.1 1.7 1.9 1.1 3.8
5.3 1.1 1.8 2.1 1.1 4.1
with the passage of time which is independent of the past
5.7 1.4 2.2 2.3 1.0 8.8
focusing only at the period of study. The NSC for any 6.9 1.5 2.7 2.5 1.1 11.8
element is calculated (Table 3) with the following 7.7 1.5 2.9 2.9 1.2 13.1
8.3 1.7 3.5 4.0 1.2 13.5
formula (Sayadi and Sayyed, 2010).
Traffic Area
concentration in the last sampling Pb Cu Cr Zn Ni Cd
concentration in first sampling 5.1 1.1 1.4 1.9 1.0 5.3
NSC = x 100
concentration in the last sampling +
5.6 1.4 1.6 1.9 1.2 6.6
concentration in first sampling 6.9 1.5 1.6 2.1 1.2 8.3
8.7 1.7 1.8 2.1 1.3 9.5
The NSC values + 100% indicates absolute 10.1 1.8 1.8 2.1 1.3 9.9
11.2 1.9 2.1 2.2 1.3 12.1
increase while-100% means absolute decrease. The value
Residential Area
of 0% can be regarded for no change in the parameters Pb Cu Cr Zn Ni Cd
under consideration. 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
Figure 1. Mean Concentrations of the heavy metals Figure 2. Industrial Area

on different sampling stations
Figure 3. Traffic Area

Figure 4. Residential Area
enrichment factor. Cadmium shows lowest concentration cumulative activity in the region. Hence the Enrichment
in soil but is has quite high enrichment factor, while factor should denote the total enrichment and or
Copper, Chromium, Zinc and Nickel shows higher metal depletion of an element and cannot evaluate the trend for
concentration but rather low EF when compared to lead. the short term accumulation.
The scatter plot of the mean concentration of When the mean values of EF and NSC for all the
heavy metals was plotted against the EF for all the three six heavy metals are studied at all the sampling stations
sampling sites (Figures 5,6,7). Per usual of the result (Figures 8, 9,10) it can be stated that Cadmium has been
showed that Zinc is having high mean concentration but enriched to a quite greater extent followed by Lead, Zinc,
it is not getting enriched in proportion to its mean Chromium, Copper and Nickel at all the sampling sites.
concentration. On the other hand Cadmium though On the other hand the Normalized Scatter Coefficient
having lowest mean concentration has higher rate of value indicates that Cadmium has got enriched in faster
enrichment. Lead shows the highest mean concentration rate at industrial area followed by Zinc, Chromium,
and also corresponding highest enrichment factor. Lead, Copper and Nickel. In traffic area Lead is getting
The behavior of Zinc may be attributed to its enriched in the faster rate followed by Cadmium,
source mainly from weathering of the parent rock while Chromium, Copper, Zinc and Nickel. But in residential
that of Cadmium and Lead mainly due to anthropogenic area, the NSC value indicate that Cu is quite enriched
activities. EF normally reveals the addition and or with the faster rate followed by Chromium, Cadmium,
removal of metal under consideration which is a result of Lead and Zinc.

Enrichment Factor
Normalized Scatter Coefficient %

Figure 5. Industrial Area Figure 8 INDUSTRIAL AREA

Enrichment Factor
Figure 6. Traffic Area Figure 9 TRAFFIC AREA

Enrichment Factor
Figure 7. Residential Area Figure 10. RESIDENTIAL AREA
CONCLUSION by Lead, Zinc, Chromium, Copper and Nickel.

The variation assessment of heavy metal Normalized Scatter Coefficient value indicate that Lead
pollution by using Enrichment Factor and Normalized is getting accumulated in a faster rate followed by
Scatter Coefficient in the soil sample collected from the Cadmium, Chromium, Copper, Zinc and Nickel. In
study area between May 2011-Oct 2011 has revealed summary the soils in the Madurai industrial, traffic and
significant increase in the six heavy metals (viz Pb, Cu, residential area are significantly contaminated by heavy
Cr, Zn, Ni and Cd). Enrichment Factor values shows metals and hence more attention to be paid to heavy
that Cadmium has enriched to a greater extent followed metal pollution particularly for Lead and Cadmium. In
Govil PK, Reddy GLN, Krishna AK. 2001.

Table 3 Normalized Scatter Coefficient (%)of the Contamination of soil due to heavy metals in Patancheru
heavy metals in the soils of the study area
Industrial Area industrial development area, Andhra Pradesh, India.
Pb Cu Cr Zn Ni Cd Environmental Geology 41:145-150.
26.1 21.9 33.8 33.8 8.6 55.4
21.1 21.1 30.9 31.7 8.1 53.2 Jarup L. 2003. Hazards of heavy metal contamination.
17.8 12.0 23.6 26.7 6.6 21.3
Brazilian Medical Bulletin 68:425-462.
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 Michalke B. 2003. Element speciation definition,
Traffic Area analytical methodology, and some examples,

Pb Cu Cr Zn Ni Cd Ecotoxicology and Environmental Safety 56:122-139.
37.0 25.9 19.5 6.6 12.5 38.7
32.9 14.6 15.8 4.9 5.7 29.1 Pekey H, Karakas D, Ayberk S. 2004. Ecological Risk
23.0 10.9 14.0 1.9 2.7 18.3
12.5 5.5 8.0 0.9 2.0 12.1 Assessment Using Trace Elements from surface
5.2 4.1 7.3 7.3 0.8 9.5 Sediments of Izmit Bay (Northeastern Marmara Sea)
0 0 0 0 0 0
Turkey. Marine Pollution Bulletin 48:946-953.
Residential Area
Pb Cu Cr Zn Ni Cd Romic M, Romic D. 2003. Heavy metal distribution in
13.2 26.4 15.3 6.1 8.9 13.9
8.8 17.9 13.1 4.7 7.2 13.4 agricultural top soils in urban area. Environmental
3.9 16.8 11.9 3.4 3.2 9.8 Geology 43(7):795-805.
2.5 1.6 8.7 2.6 2.5 6.1
0.8 0.6 5.4 0.4 0.6 5.6
Sarala thambavani D and vathana.m. 2012. Ambient
0 0 0 0 0 0
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from the Madurai city and to maintain the ecological
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quality criteria, a need to be taken. Sayadi MH, Sayyed MRG. 2010. Comparative
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Journal of Research in Biology An International Open Access Research Journal

Anatomical variation in the olfactory apparatus of marine teleosts

Phone No: Article Citation:

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

INTRODUCTION and Platycephalus scaber (Linnaeus, 1758), an estuarine

Figure 1A - Schematic representation of Rastrelliger kanagurta.

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

Figure 2A - Schematic representation of Scomberoides commersonianus.

Figure 3A - Schematic representation of Platycephalus scaber.

Journal of Research in Biology (2013) 3(1): 742-748 746

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

Kleerekoper H. 1969. Olfaction in Fishes. Indiana

Knouff RA. 1935. The developmental pattern of

Kotrschal K, Krautgartner W, Hansen A. 1997.

Munday PL, Dixson DL, Donelson JM, Jones GP,

Nevitt GA. 1991. Do fish sniff? A new mechanism of

Ojha PP and Kapoor AS. 1973. The anatomy of

Teichmann H, 1954. Vergle chende ntersuchungen an Advantages

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

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

Corresponding author: Article Citation:

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

INTRODUCTION pies and porridges. Nevertheless, this species has

Journal of Research in Biology (2013) 3(1): 747-758 749

Table 1. Purple yam (D. trifida) incorporated

Caloric value (kcal/100 g)

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

Centesimal composition analyses of purple yam

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

Carbohydrate and caloric values were determined

Table 2. Yam centesimal composition.

Lipid (%) 0.20 0.10 1.13 0.69

Protein (%) 2.00 2.30 1.83 0.13

Journal of Research in Biology (2013) 3(1): 747-758 751

followed by post hoc tests (Table 3), it may be asserted 80

composition and caloric values of the four purple yam 50

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

20% neither modifies bread centesimal composition nor 40 80 160 330

caloric value. As for lipid, statistically significant

Journal of Research in Biology (2013) 3(1): 747-758 753

0.4 15% Bread

et al., 2008). Corroborating data from Hsu et al., (2004), 70

it was confirmed that the antioxidant activity rose as the 60

observed by Hsu et al., (2004) in flour of Taiwan purple 20

yam (D. purpurea), was also detected in the Amazonian 10

Antioxidant activity determination through the lipid

As it was observed by the DPPH method, the

distribution normality of the three tested purple yam

homocedasticity among the formulations through the

Levene test on all sensory attributes evaluated in the

acceptance test (i.e. statistically significant values at 0.6285

Friedman ANOVA followed by post hoc tests

(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

Choosing a determined food should depend

Journal of Research in Biology (2013) 3(1): 747-758 755

Table 7. Purple yam incorporated breads microbiological analysis.

mainly on its nutritional value. Nevertheless, color, ACKNOWLEDGEMENTS

756 Journal of Research in Biology (2013) 3(1): 747-758

Journal of Research in Biology (2013) 3(1): 747-758 757

Omonigho SE and Ikenebomeh MJ. 2000. Effect of