Journal of Research in Biology Volume 3 Issue 6

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Department of Pathology, Army Medical College, Rawalpindi, Pakistan.

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University of Kordofan, Elobeid-SUDAN.

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Central Institute of Medicinal and Aromatic Plants, Lucknow, India.

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C.S.J.M. University, India.

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Federal University of Rondnia, Brazil.

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Stefan cel Mare University of Suceava, Romania.

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Govt. Dungar College, Bikaner.

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School of Life Sciences, Bharathidasan University, India.

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Arts and commerce girls college Raipur (C.G.), India.

Suwattana Pruksasri [Enzyme technology, Biochemical Engineering]
Silpakorn University, Thailand.

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Yogeshwari Mahavidyalaya, Ambajogai, India.

Dr. Pankaj Sah [Environmental Science, Plant Ecology]
Higher College of Technology (HCT), Al-Khuwair.

Dr. Erkan Kalipci [Environmental Engineering]
Selcuk University, Turkey.

Dr Gajendra Pandurang Jagtap [Plant Pathology]
College of Agriculture, India.

Dr. Arun M. Chilke [Biochemistry, Enzymology, Histochemistry]
Shree Shivaji Arts, Commerce & Science College, India.

Dr. AC. Tangavelou [Biodiversity, Plant Taxonomy]
Bio-Science Research Foundation, India.

Nasroallah Moradi Kor [Animal Science]
Razi University of Agricultural Sciences and Natural Resources, Iran

T. Badal Singh [plant tissue culture]
Panjab University, India

Dr. Kalyan Chakraborti [Agriculture, Pomology, horticulture]
AICRP on Sub-Tropical Fruits, Bidhan Chandra Krishi Viswavidyalaya,
Kalyani, Nadia, West Bengal, India.

Dr. Monanjali Bandyopadhyay [Farmlore, Traditional and indigenous
practices, Ethno botany]
V. C., Vidyasagar University, Midnapore.

M.Sugumaran [Phytochemistry]
Adhiparasakthi College of Pharmacy, Melmaruvathur, Kancheepuram District.

Prashanth N S [Public health, Medicine]
Institute of Public Health, Bangalore.

Tariq Aftab
Department of Botany, Aligarh Muslim University, Aligarh, India.

Manzoor Ahmad Shah
Department of Botany, University of Kashmir, Srinagar, India.

Syampungani Stephen
School of Natural Resources, Copperbelt University, Kitwe, Zambia.

Iheanyi Omezuruike OKONKO
Department of Biochemistry & Microbiology, Lead City University,
Ibadan, Nigeria.

Sharangouda Patil
Toxicology Laboratory, Bioenergetics & Environmental Sciences Division,
National Institue of Animal Nutrition
and Physiology (NIANP, ICAR), Adugodi, Bangalore.

Jayapal
Nandyal, Kurnool, Andrapradesh, India.

T.S. Pathan [Aquatic toxicology and Fish biology]
Department of Zoology, Kalikadevi Senior College, Shirur, India.

Aparna Sarkar [Physiology and biochemistry]
Amity Institute of Physiotherapy, Amity campus, Noida, INDIA.

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Department of Physiology, University of Calcutta, Kolkata, INDIA .

Maruthi [Plant Biotechnology]
Dept of Biotechnology, SDM College (Autonomous),
Ujire Dakshina Kannada, India.

Veeranna [Biotechnology]
Dept of Biotechnology, SDM College (Autonomous),
Ujire Dakshina Kannada, India.

RAVI [Biotechnology & Bioinformatics]
Department of Botany, Government Arts College, Coimbatore, India.

Sadanand Mallappa Yamakanamardi [Zoology]
Department of Zoology, University of Mysore, Mysore, India.

Anoop Das [Ornithologist]
Research Department of Zoology, MES Mampad College, Kerala, India.

Dr. Satish Ambadas Bhalerao [Environmental Botany]
Wilson College, Mumbai

Rafael Gomez Kosky [Plant Biotechnology]
Instituto de Biotecnologa de las Plantas, Universidad Central de Las Villas
Eudriano Costa [Aquatic Bioecology]
IOUSP - Instituto Oceanogrfico da Universidade de So Paulo, Brasil

M. Bubesh Guptha [Wildlife Biologist]
Wildlife Management Circle (WLMC), India

Rajib Roychowdhury [Plant science]
Centre for biotechnology visva-bharati, India.

Dr. S.M.Gopinath [Environmental Biotechnology]
Acharya Institute of Technology, Bangalore.

Dr. U.S. Mahadeva Rao [Bio Chemistry]
Universiti Sultan Zainal Abidin, Malaysia.

Hrida Regina Nunes Salgado [Pharmacist]
Unesp - Universidade Estadual Paulista, Brazil

Mandava Venkata Basaveswara Rao [Chemistry]
Krishna University, India.

Dr. Mostafa Mohamed Rady [Agricultural Sciences]
Fayoum University, Egypt.

Dr. Hazim Jabbar Shah Ali [Poultry Science]
College of Agriculture, University of Baghdad , Iraq.

Danial Kahrizi [Plant Biotechnology, Plant Breeding,Genetics]
Agronomy and Plant Breeding Dept., Razi University, Iran

Dr. Houhun LI [Systematics of Microlepidoptera, Zoogeography, Coevolution,
Forest protection]
College of Life Sciences, Nankai University, China.

Mara de la Concepcin Garca Aguilar [Biology]
Center for Scientific Research and Higher Education of Ensenada, B. C., Mexico

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

Dr. Pritam Chattopadhyay [Agricultural Biotech, Food Biotech, Plant Biotech]
Visva-Bharati (a Central University), India

Dr. Preetham Elumalai [Biochemistry and Immunology] Institute for
Immunology Uniklinikum, Regensburg, Germany

Dr. Mrs. Sreeja Lakshmi PV [Biochemistry and Cell Biology]
University of Regensburg, Germany

Dr. Alma Rus [Experimental Biology]
University of jan, Spain.

Dr. Milan S. Stankovi [Biology, Plant Science]
University of Kragujevac, Serbia.

Dr. Manoranjan chakraborty [Mycology and plant pathology]
Bishnupur ramananda college, India.

Table of Contents (Volume 3 - Issue 6)

Serial No Accession No Title of the article Page No

1 RA0363 Diversity of Wetland dependent birds around the Bhadra Reservoir
Project (BRP) area, Karnataka.
Dayananda GY.

1054-1059
2 RA0374 Preliminary investigations on quantity and proximate quality of
maggots produced from four different sources of livestock wastes.

Afamdi Anene, Olivia C. Afam-Anene, Kelechi Ike and Nnamdi H.
Ekekwe.
1060-1065

3

RA0297

Recent biophysical characteristics of domestic water sources in Owerri
Metropolis, Nigeria.

Nwachukwu MI, Eziuzor SC, Duru MKC, Nwachukwu IO, Ukaga CN,
Udujih OS and Udujih GO.

1066-1071
4 RA0340 Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae
and the moth, Philosamia ricini.

Bendang Ao and Sentimenla.

1072-1085

Article Citation:
Dayananda GY.
Diversity of Wetland dependent birds around the Bhadra Reservoir Project (BRP)
area, Karnataka.
Journal of Research in Biology (2013) 3(6): 1054-1059
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Diversity of Wetland dependent birds around the Bhadra Reservoir
Project (BRP) area, Karnataka
Keywords:
Wetland birds, diversity, wetlands, Bhadra Reservoir Project .
ABSTRACT:

The study of bird species inhabiting certain wetlands around Bhadra
Reservoir Project (BRP), Shivamogga, Karnataka was carried out from February 2008
to January 2010. The total of 68 species of wetland birds belonging to nineteen
families and six orders. Of these, Anatidae (15%) and Ardidae (13%) have more than
nine species. The diversity may be attributed the moderate volume of water storage,
availability of food and assured protection to these birds. Additionally we recorded
seven types of migratory birds visiting these ponds. Those include White-necked Stork,
Shoveler, Pintail, Grey Plover, Curlew, Ringtailed-fishing Eagle and Black-winged Stilt.
All these wetlands are important places for foraging activity of wetland birds. In order
to protect these wetland birds, the wetlands should be conserved by controlling
encroachment, pollution and other anthropogenic activities.
1054-1059 | JRB | 2013 | Vol 3 | No 6
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.
www.jresearchbiology.com
Journal of Research in Biology
An International
Scientific Research Journal
Authors:
Dayananda GY.

Institution:
Department of P.G. Studies
and Research in Applied
Zoology, Bioscience
Complex, Jnana Sahyadri,
Kuvempu University,
Shankaraghatta 577 451.
Shimoga.

Corresponding author:
Dayananda GY.

Email Id:

Web Address:
http://jresearchbiology.com/
documents/RA0363.pdf.
Dates:
Received: 06 July 2013 Accepted: 22 July 2013 Published: 04 Sep 2013
An International Scientific Research Journal
Original Research

INTRODUCTION
Wetlands are the treasures of avifaunal species
richness and these are the important ecological
significance areas, which serves as a major link between
the natural resources and agricultural practices. Wetlands
of lentic group form a favorable habitat to various groups
of animals especially waterfowl, that need food, water
for drinking, wallowing and abode. Wetlands are known
to be most productive and diverse ecosystems on the
earth. Water birds are perhaps the most visible
manifestation of faunal diversity but many other groups
also inhabit these wetlands. Wetlands are fragile
ecosystems, which are fast deteriorating and shrinking
due to man made activities. India has 65,000 wetlands
covering an area of 4.5 million hectares (Anon, 1990).
The diversity of water birds obviously indicate the
quality and healthy condition of the ecosystem in the
country. Concerning the realm of this study, some other
works have been carried out by Dayananda (2009);
Nanda et al., (2010); Rajpar and Zakaria (2010); Mohsen
et al., (2011). The aim of this study is to assess the
diversity of wetland birds in and around Bhadra
Reservoir Project area.

MATERIALS AND METHODS
The checklist of wetland birds around the BRP
area was made by sighting the birds with 8 x 50
binoculars. The field guides (Ali, 1996; Sonobe and
Usui, 1993; Inskipp and Inskipp, 1991; Fleming et al.,
2000; Kazmierczak and Perlo, 2000; Grimmett et al.,
2001) were used for bird identification. The wetland bird
census was conducted in morning hours from 06:00 AM
to 10:00 AM and evening 04:00 PM to 06:00 PM by
walking. Study of wetland birds around the BRP area
was carried out from February 2008 to January 2010,
every month at regular interval by direct counting
method (Colin et al., 1993; William, 1997). The
residential status and abundance criteria was calculated
using presence and absence scoring method and then
percentage of birds occurrence was calculated to
determine the status. The modified score classes used on
the basis of total bird recorded during study period i.e.,
1-5%= rare (R), 6-10%=Uncommon (UC), 11-13%=
common (C) and >14% = Verycommon (VC) as
accomplished by Mc Kinnon and Philips (1993).

RESULTS AND DISCUSSION
A total of 68 species of birds were found
associated with the Bhadra Reservoir. Of which 40
species are resident, 21 residents with local migratory
and seven are migratory. Some of the migratory birds
recorded includes White-necked Stork, Shoveler, Pintail,
Grey Plover, Curlew, Ringtailed-fishing Eagle and Black
-winged Stilt. These are winter migrants used the
wetlands for foraging, resting and other activities till
favorable condition of their native and some residential
wetland birds such as the herons, egrets, ibises and storks
used the trees and shurbs as roosting site. These species
were found during the study period on the ground
feeding of fishes, amphibians and crutaceans. The report
suggested that the wetlands are important places for
foraging of wetland birds. This observation got support
from earlier publications which reported that, habitat has
long been used as a predictor of bird species abundance
and variety of birds has developed different preferences
for habitat (Huston, 1994; Lameed, 2011). During study
68 bird species belonging to 19 families and six orders
were found on the wetland (Table-1). The status based
upon percent occurrence of bird species representing
different families with respect to total bird species
presently recorded was Anatidae (14.71) > Ardeidae
(13.24) > Charadriidae (10.29) > Alcedinidae (7.35) =
Motacillidae (7.35) > Rallidae (5.88) = Jacanidae (5.88)
= Threskiornithidae (5.88) > Accipitridae (4.41) >
Phalacrocoracidae (2.94) = Ciconiidae (2.94) =
Scolopacidae (2.94 ) = Laridae (2.94 ) = Alaudidae
(2.94) = Corvidae (2.94) = Ploceidae (2.94)
>Podicipedidae (1.47) = Recurvirostridae (1.47) =
Dayananda, 2013
1055 Journal of Research in Biology (2013) 3(6): 1054-1059

Dayananda, 2013
Journal of Research in Biology (2013) 3(6): 1054-1059 1056
Table 1. Wetland bird diversity around the Bhadra Reservoir Project Area
Sl.
No
Order Family Common Name Scientific Name RS AS FH
Podicipediformes Podicipedidae Little Grebe Tachybaptus ruficollis R C P
Pelecaniformes Phalacrocoracidae Little Cormorant Phalacrocorax niger RM VC P
Oriental Darter Anhinga melanogaster RM UC P
Ciconiiformes Ardeidae Grey Heron Ardea cinerea RM C P
Purple Heron Ardea purpurea RM C P
Pond Heron Ardeola grayii R VC P
Night Heron Nycticorax nycticorax R UC P
Cattle Egret Bubulcus ibis R VC P
Large Egret Casmerodius albus RM VC P
Median Egret Mesophoyex intermedia R VC P
Little Egret Egretta garzetta R VC P
Chestnut Bittern Ixobrychus cinnamomeus RM UC P
Threskiornithidae Black-headed Ibis Threskiornis melanocephalus R VC P
Black Ibis Pseudibis papillosa RM C P
Eurasian Spoonbill Platalea leucorodia RM R P
Glossy Ibis Plegadis falcinellus RM C P
Ciconiidae White-necked Stork Ciconia nigra M R P
Open-bill Stork Anastomus oscitans R UC P
Anseriformes Anatidae Lesser-whistling Teal Dendrocygna javanica R C O
Common Teal Anas crecca RM C O
Spot-billed Duck Anas poecilorhyncha RM VC O
Garganey Anas querquedula R UC O
Nakta or Comb Duck Sarkidiornis melanotos R UC O
Shoveler Anas clypeata M R O
Cotton Teal Nettapus coromandelianus R VC O
Mallard Anas platyrhynchos RM UC O
Pintail Anus acuta M R O
Brahminy Duck Tadorna ferruginea RM UC O
Accipitridae Common Pariah Kite Milvus migrans R VC C
Brahminy Kite Haliastur indus R VC C
Ring tailed fishing Eagle Haliaeetus leucoryphus M R C
Gruiformes Rallidae White-breasted Water
hen
Amaurornis phoenicurus R VC I,G
Indian Moorhen Gallinula chloropus R VC O
Purple Moorhen Porphyrio porphyrio R VC O
Common Coot Fulica atra R VC O

Sturnidae (1.47) (Fig. 1). The Anatidae and Ardeidae had
more than nine species, this can be considered as good
indicators of the health of these wetlands.

The diversity may be attributed the moderate
volume of water storage, availability of food sources
(fish, crustaceans, invertebrates, aquatic plants and
plankters), shelter and assured protection to these birds.
Dayananda, 2013
Charadriiformes Jacanidae
Bronze-winged Jacana
Metopidius indicus R VC I/G

Pheasant-tailed Jacana
Hydrophasianus chirurgus RM VC I/G

Brown Crake
Amaurornis akool R C I

Water Cock or Kora
Gallicrex cinerea LM C I

Charadriidae
Red-wattled Lapwing
Vanellus indicus R VC I

Yellow-wattled Lapwing Vanellus malabaricus R VC I

Little-ringed Plover Charadrius dubius RM C I

Grey Plover Pluvialis squatarola M R I

Curlew Numenius arquata M R I

Common Sandpiper
Actitis hypoleucos RM VC I

Marsh Sandpiper
Tringa stagnatilis R C I

Recurvirostridae Black-winged Stilt Himantopus himantopus M R I

Scolopacidae
Painted Snipe
Rostratula benghalensis R C I

Common Snipe
Gallinago gallinago RM C I

Laridae
Indian River Tern
Sterna aurantia R C P

Common Tern
Sterna hirundo RM C P

Alcedinidae
Lesser-pied Kingfisher
Ceryle rudis R C P

Small-blue Kingfisher
Alcedo atthis R C P

Blue-eared Kingfisher
Alcedo meninting R C P

White-breasted King-
fisher
Halcyon smyrnensis R VC P

Stork-billed Kingfisher
Pelargopsis capensis R C P

Alaudidae
Crested Lark
Galerida cristata R C I

Black-bellied Finchlark Eremopterix griseus R UC I

Sturnidae Indian Myna Acridotheres tristis R VC I

Corvidae House Crow Corvus splendens R VC O

Jungle Crow Corvus macrorhynchos R VC O

Motacillidae Large pied Wagtail Motacilla maderaspatensis R C I

White Wagtail Motacilla alba RM VC I

Yellow Wagtail Motacilla flava R C I

Yellow-headed Wagtail Motacilla citreola RM C I

Paddy Field Pipit Anthus novaeseelandiae R VC I

Ploceidae Baya weaver bird Ploceus philippinus R VC I

Black breasted weaver
bird
Ploceus benghalensis R VC I
Common and Scientific names are as followed by Manakadan and Pittie, 2001.
RS Residential Status of the birds: R Resident, M Migratory, RM Resident with migratory. AS Abundance
Status of the birds: R Rare, UC Uncommon, C Common, VC Verycommon. FH Food habit of the birds:
I Insectivore; P- Piscivore; O-Omnivore; I/G Insectivore with Grainivore.
The family Anatidae dominated the list with ten species,
which was represented 14.71% of the total number of
wetland birds present in the study area. Ardeidae was
represented by nine species with a relative abundance of
13.24%. Charadriidae was represented by seven species.
Motacillidae and Alcedinidae were represented by five
species. Threskiornithidae, Rallidae, Jacanidae were
represented by four species. Accipitridae was represented
by three species and Phalacrocoracidae, Ciconiidae,
Scolopacidae, Laridae, Alaudidae, Corvidae and
Ploceidae were represented by two species each whereas
Podicipedidae, Recurvirostridae and Sturnidae had single
species each. Among the birds recorded in this study,
about 36.76 % (25 species) are both piscivores and
insectivores and 22.06 % (15 species) are omnivores and
4.41 % (3 species) are carnivores respectively (Fig. 1).
In the present study, the analysis on the status
shows that twenty five species are common, twenty eight
species very common, nine species uncommon and eight
species rare. The abundance of birds may be influenced
by availability food for birds in the form of plants,
vertebrates and invertebrates, some of them feed in
wetland soil, water column and dry landscape in and
around the wetlands. The present work is in conformity
with the earlier work of Dayananda (2008) carried out in
Ramanakere of Davanagere district. Similarly, this
results were in agreement with the earlier works of
Rajashekara and Venkatesha (2011); Lameed, 2011;
Bhatnagar et al., (2008) who also reported the varying
diversity of birds in different lakes due to different
habitat conditions for roosting, nesting, feeding and
availability of food sources.

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Fig. 1. Percent composition of avian families represented by species
richness of waterbirds around BRP area

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Dayananda, 2013
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Article Citation:
Afamdi Anene, Olivia C. Afam-Anene, Kelechi Ike and Nnamdi H. Ekekwe
Preliminary investigations on quantity and proximate quality of maggots produced
from four different sources of livestock wastes.
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Preliminary investigations on quantity and proximate quality of maggots
produced from four different sources of livestock wastes
Keywords:
maggots, proximate quality, livestock wastes
ABSTRACT:

Maggot, housefly larva was grown on four substrates namely: poultry (layer)
droppings, cattle dung, pig dung, and whole cattle blood. Poultry droppings produced
maggots with the highest wet and dry weight, while the lowest weights were recorded
for pig dung. The values ranged between 58.73g and 8.18g for wet weight and 12.79g
and 2.97g for dry weight respectively. Proximate compositions of the maggots were
determined using standard methods. Results indicate that the crude protein content
of the maggots ranged from 55.4% in maggots grown on pig dung to 57.42% in
maggots grown on cattle blood. The crude fibre contents ranged between 0.32% and
0.21%. Maggots produced from pig dung and cattle blood recorded the highest ash
content and the values were 11.09% and 11.20% respectively. Moisture content was
highest (10.14%) for maggots produced from cattle dung. Fat content of the maggots
produced from the different livestock wastes ranged between 21.06% and 22.66%.
Significant differences (p<0.05) in the proximate composition of the maggots were
only observed in the crude fiber, ash and moisture contents. The results from this
study showed that the substrates used can produce substantial quantities of maggots
with varying degrees of success.
1060-1065 | JRB | 2013 | Vol 3 | No 6

An International
Authors:
Afamdi Anene
1
, Olivia C.
Afam-Anene
2
, Kelechi Ike
1

and Nnamdi H. Ekekwe
1

Institution:
1. Animal Nutrition
Laboratory, Department of
Animal Science/Fisheries,
Abia State University,
Umuahia Campus. Abia
State, Nigeria.

2. Department of Nutrition
and Dietetics Imo State
University, P. M. B. 2000,
Owerri, Imo State, Nigeria.

Afamdi Anene.

Email Id:

Web Address:
Dates:
Received: 07 Aug 2013 Accepted: 12 Aug 2013 Published: 18 Oct 2013
Original Research

INTRODUCTION
Feed is known to be the single most expensive
factor in animal and aquaculture production of which
protein is the feed constituent with the highest cost
implications (Aniebo et al., 2008). Plant protein sources
as alternative non-conventional protein have their
limitations largely due to the presence of secondary
metabolites such as alkaloids, glycosides, oxalic acids,
phytates, protease inhibitors, haematoglutinin, saponins,
cyanoglycosides and linamarin etc to mention a few.
Plant protein sources have the advantage of low cost
implications as well as rich nutrient levels (Sogbesan,
2006, Sogbesan et al., 2006). These anti-nutritional
factors negate growth and other physiological activities
at higher inclusion levels (Oresegun and Alegbeleye,
2001). Fish meal which is the guaranteed protein feed
ingredient in animal diets and it costs as much as
$2.1 per kilogram, approximately N300/kg which is
about thrice the cost of soya bean meal and four times
the cost of groundnut cake (GNC) (Aniebo et al., 2008).
Consequently there is a drive to develop other protein
sources too. Maggot meal has been reported to possess
good nutritional value, cheaper and less tedious to
produce than most other sources of animal protein.
Housefly maggots have been used as protein ingredients
in fish feeds (Aniebo et al., 2008), poultry feeds (Inaoka
et al., 1999, Adeniji, 2007, Hwangbo et al., 2009) and
crustaceans (Cao et al., 2012).
The housefly (Musca domestica Linnaeus 1758)
is the most common fly species and belongs to the
phylum Insect and order Diptera. The larval forms
(maggots) of houseflies feed on decaying organic matter
thereby giving them the ability to degrade wastes into
valuable biomass that are nutrient (fat, protein etc) rich.
Many studies (Akpodiete et al., 1993, Awoniyi and
Aletor, 2002, Teguia 2005, Aniebo et al., 2008) have
been conducted on the production of housefly biomass in
simulated environments with a view in utilizing such as
feed for farm animals.
Pig manure, wheat bran, cattle gut and rumen
contents, fish guts and cattle blood are some of the
substrates that have been reportedly used for the
production of maggots (Viroje et al., 1988; Ekoue et al.,
2000; Aniebo et al., 2008; Ossey et al., 2012; Zhu et al.,
2012). However, there is a lacuna of information on the
comparative advantage in quantity of production of these
substrates. There is a dearth of information on the
production potentials of different substrates for the
production of maggots.
This study is aimed at a comparative evaluation of;
The quantity of maggots harvested from poultry
droppings, pig dung, cattle dung and cattle blood,
without any additional fly attractants and without
absorbents,
The proximate quality of the maggot so produced
from these livestock wastes (substrates).

The experiment was carried out at the Teaching
and Research Farm, Abia State University, Umuahia
Location. The treatments consisted of 30 kg each of
poultry droppings, cattle and pig dung; and congealed
blood. These were replicated three times giving each
replicate a weight of 10 kg and randomly placed in a
roofed open space. The exposed substrates attracted
houseflies which laid eggs that hatched into larva called
maggots. Each substrate was sprinkled with half a liter of
untreated borehole water for a period of four days to
prevent desiccation.
Harvesting
Harvesting was done on the 4
th
day using the
sedimentation technique. Each replicate was mixed with
7-10 liters of water and allowed to stand for 10 minutes
to completely separate the maggots from the substrates.
Upon mixing, the substrates sank while the maggots
floated and were collected using a 3mm sieve. Harvested
maggots were taken to the laboratory for weight
measurements and chemical analyses.
Anene et al., 2013
Data Collection, Sample and Data Analysis
Maggots from each replicate were weighed to the
nearest 0.1g when wet and then weighed after drying to a
constant weight at 35
o
C in an oven using a digital
weighing balance. Dried maggots from each treatment
were blended into a smooth paste in a 3.8 L kitchen-type
blender (Warning Products, New Hartford, CT) which
was thoroughly cleaned and dried between samples.
Triplicate determination was made for each treatment.
Samples were analysed for crude protein (CP), crude
fiber (CF), ash, nitrogen free extract (NFE), and moisture
using methods described by AOAC (1995). All data were
subjected to Analysis of Variance (ANOVA) using SPSS
version 17 and differences in means were separated
using Duncans Multiple Range Test (Duncan, 1955).

RESULTS AND DISCUSSIONS
The wet and dry weights of maggots produced
from the four different wastes are presented in Table 1.
The result from this study shows that 1kg each of poultry
manure, pig dung, cattle dung and congealed blood
yielded a mean wet weight of 58.73, 8.18, 12.92 and
21.77 g of maggot. Similarly, the dry weight of maggot
yield from the 1kg of the four substrates were 12.7 g
from poultry droppings, 2.97 g from pig dung, 4.18 from
cattle dung and 7.79 g from congealed cattle blood.
These results showed that there were significant
differences (p>0.05) in the weights of maggots (wet and
dry) produced from the wastes. The trend in the quantity
of maggot production was as follows: Poultry droppings
> Cattle blood > Cattle dung > Pig dung. Insects have
been shown to exhibit marked preferences for particular
substrates for oviposition (Zvereva and Zhemchuzhina,
1988). Similarly, sites for oviposition can be influenced
by many factors among which are moisture, nutritive
value of the substrate and the presence or absence of an
oviposition attractant. In this study poultry manure
characterized by high ammonium levels produced the
highest quantity of manure. (Pastor et al., 2011) have
shown that ammonia, is an effective oviposition
attractant.
The results obtained in this study compared
favorably with some literature reports on maggot
production (Akpodiete et al., 1993, Awoniyi and Aletor,
2002). It is important to note that the quantities of
maggot produced in this study were generally lower than
those reported in Aniebo et al., (2008). Aniebo et al.,
(2008) used absorbent material namely wheat brain, rice
dust and saw dust and these may have accounted for by
the higher harvests of maggots. These report however
(Akpodiete et al., 1993, Awoniyi and Aletor, 2002,
Aniebo et al., 2008) agree that the quantity of maggot
produced was primarily dependent on the nature of the
substrate.
Other factors such as moisture control and
inadequate aeration of substrates may influence the
quantity of maggot yield from the substrates (Calvert
et al., 1971). Aniebo et al., 2008 reported that high
density of substrates decreased aerobic conditions which
could adversely affect the development and survival of
both of eggs and hatched larvae.
Table 2 summarizes the proximate composition
of maggots produced from the different livestock wastes.
Crude protein content ranged between 55.54% in
maggots produced from pig dung to 56.25% in maggots
produced from poultry droppings and did not indicate
any significant difference (p>0.05) amongst the various
Anene et al., 2013
Treatments
Mean Yield (g)
per Kg (N=3)
Wet Weight Dry Weight
Poultry Droppings 58.7300.34
a
12.790.22
a

Pig dung 08.1800.22
d
02.970.17
d

Cattle dung 12.9200.16
c
04.180.52
c

Cattle Blood 21.7700.31
b
07.790.41
b

Table 1: Weight of maggots produced from
different livestock wastes
Means in the same column with different superscripts
are significantly different (p<0.05).

substrates. The crude protein content of housefly
maggots has been shown by various workers to vary
between 40 and 60% (Inaoka et al., 1999, Heuz and
Tran; 2013). Hwangbo et al., (2009) recorded a protein
content of 63.99% in maggots grown on chicken
droppings sprinkled with powdered milk and sugar.
Lower protein regimes of 45% - 48% were reported by
Fasakin et al., (2003). It is possible that higher protein
values in maggots may be attributed to the higher
nutritional content of the substrate.
Table 2 also shows the ether extract content of
maggots produced from various substrates. This
parameter ranges from 27.06-22.66% and did not vary
significantly (p>0.05) with the substrate type. Inaoka
et al., (1999) recorded crude fat content of 20% in
maggots while some other authors have reported a highly
variable lipid contents ranging between 9-25% (Heuz
and Tran; 2013). The results of this study on the fat
content of maggot produced from different substrates
were in tandem with those of other authors. Drying
methods (sun drying and oven drying) have been shown
to influence the ratio of protein to fat ratio (Aniebo and
Owen, 2010). Heuz and Tran (2013) observed that fatty
acid profiles of maggots are largely influenced by the
substrates on which they are grown and this may account
for the high variability in fat content reported by various
authors (Inaoka et al., 1999, Hwangbo et al., 2009,
Aniebo and Owen, 2010).

There were significant differences (p<0.05) in
ash content of maggots reared on various substrates. Ash
content of maggots reared on pig dung was 11.09% and
those reared on cattle blood was 11.20%. These values
were significantly lower (p<0.05) than the ash content of
maggots reared on poultry manure (10.8%) and pig dung
(11.09%). These results on ash content of maggots differ
from a value of 2.74% reported for larvae of dung beetle
(Aphodius rufipes) (Paiko et al., 2012) but are in tandem
with those published by Hwangbo et al., (2009). Ash
content is an indication of the mineral content of feed
materials.
The crude fiber content of the maggots from all
the substrates were all less than 1%. Similarly, there
were significant differences in the crude fiber content.
These low values indicate that maggot meal is not a good
source of fiber. Similar low values ranging between
0.16% for cattle blood and 0.61% for pig dung were
recorded for nitrogen free extracts (NFE). There were no
significant differences (p<0.05) in the values obtained
for this parameter.

CONCLUSION AND RECOMMENDATION
In this study, maggots of housefly larvae were
grown on four substrates namely: poultry (layer)
droppings, cattle dung, pig dung, and whole cattle blood
in a roofed open space. The findings from this
experiment showed that poultry droppings produced
maggots with the highest wet and dry weights and this
Anene et al., 2013
Treatments
Parameters Poultry droppings Pig dung Cattle Dung Cattle Blood
Crude Protein 56.250.21
a
55.54 0.15
a
56.00
a
0.00 57.42
a
0.00
Crude fibre 00.320.08
a
00.260.05
ab
00.21 0.01
b
00.290.06
ab

Ash 10.800.17
b
11.090.15
a
10.90 0.12
b
11.200.11
a

Ether Extract 22.320.09
a
22.640.07
a
22.66 0.21
a
21.060.19
a

Nitrogen Free Extract 00.170.04
a
00.610.07
a
00.07 0.01
a
00.160.07
a

Moisture content 10.120.11
b
09.840.12
b
10.14 0.21
a
09.860.16
b

Table 2: Proximate composition of maggots produced from different livestock wastes
*
abc
: Means along the same row with different superscripts are significant (p<0.05).
result may be due to the presence of ammonia in poultry
dropping. This study further strengthens the observation
that the quantity of maggot produced by a substrate is
primarily dependent on the nature of the substrate.
With the exception of the crude protein and fat
contents, the ash, nitrogen free extract and moisture
composition were affected by the type of substrate used
in the study. The protein content in the maggots
produced from poultry (layer) droppings, cattle dung, pig
dung, and whole cattle blood were comparable to
literature reports on maggots grown on other substrates.
The high protein content in the maggots would greatly
encourage and promote livestock production and fish
production bringing about economic affordability of the
much needed animal protein. The results also show that
maggot meal is not a good source of fiber. This study
also further strengthens the role of maggots in
biodegradation of livestock/animal wastes and its
importance in the management of wastes in the industry.
In all, this work has provided vital information on the
chemical composition of maggot meal which would
facilitate its incorporation into animal and fish feeds.

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Aniebo AO and Owen OJ. 2010. Effects of Age and
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Awoniyi TAM and Aletor VA. 2002. Proc. 29th Ann.
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Calvert CC, Martins RD and Eby HJ. 1971. Housefly
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Cao JunMing, Yan Jing, Huang YanHua, Wang
GuoXia, Zhang RongBin, Chen XiaoYing, Wen
YuanHong, Zhou TingTing. 2012. Effects of
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Fasakin EA, Balogun AM and Ajayi OO. 2003.
Evaluation of full-fat and defatted maggot meals in the
feeding of clariid catfish Clarias gariepinus fingerlings.
Aquaculture Research. 34(9): 733-738.

Heuz V and Tran G. 2013. Housefly maggot meal.
Feedipedia.org. A programme by INRA, CIRAD, AFZ
and FAO. http://www.feedipedia.org/node/671.

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Hwangbo J, Hong EC, Jang A, Kang HK, Oh JS,
Kim BW and Park BS. 2009. Utilization of house fly
maggots a feed supplement in the production of broiler
chickens. Journal of Environmental Biology. 30(4): 609 -
614.

Inaoka T, Okubo G, Yokota M, Takemasa M. 1999.
Nutritive Value of House Fly Larvae and Pupae Fed on
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Oresegun A and Alegbeleye WO. 2002. Serum and
Tissue Thiocyanate concentration in Tilapia
(Oreochromis niloticus) Fed Cassava Peel Based Diets
supplemented with DL Methionine. Journal of
Aquaculture in the Tropics.17(2): 93-100.

Ossey YB, Koumi AR, Koffi KM, Atse BC, Kouame
LP. 2012. Use of soybean, bovine brain and maggot as
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(1): 2099-2108.

Paiko YB, Dauda BEN, Salau RB and Jacob JO.
2012. Preliminary Data On The Nutritional Potentials of
The Larvae Of Edible Dung Beetle Consumed In Paikoro
Local Government Area Of Niger State, Nigeria.
Continental J. Food Science and Technology 6(2):38- 42.

Pastor B, ikov H, Koznek M, Martnez-Snchez
A, Tak P and Rojo S. 2011. Effect of the size of the
pupae, adult diet, oviposition substrate and adult
population density on egg production in Musca
domestica (Diptera: Muscidae). Eur. J. Entomol., 108
(4): 587596.

Sogbesan OA. 2006. Effects of different organic
substrates on growth and survival of long winged termite
(Macrotermes subhyabrius) under laboratory conditions.
Afr. J. Gen. Agric., 2(2): 37-44.

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2006. Harvesting techniques and evaluation of maggot
meal as animal dietary protein source for Heteoclarias
in outdoor concrete tanks. World J. Agric. Sci., 2 (4):
394-402.

Teguia A and Beynen AC. 2005. Alternative feedstuffs
for broilers in Cameroon. Livestock Research for
Rural Development 17 (3). http://www.lrrd.org/lrrd17/3/
tegu17034.htm.

Viroje W and Malin S. 1989. Effects of fly larval meal
grown on pig manure as a source of protein in early
weaned pig diets. Thurakit Ahan Sat, 6 (21): 25-31.

Zhu FX, Wang WP, Hong CL, Feng MG, Xue ZY,
Chen XY, Yao YL, Yu M. 2012. Rapid production of
maggots as feed supplement and organic fertilizer by the
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Anene et al., 2013

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Recent biophysical characteristics of domestic water
sources in Owerri Metropolis, Nigeria.
Keywords:
Bio-load, biophysical characteristics, infections, water sources, Owerri metropolis.
ABSTRACT:

The recent biophysical characteristics of domestic water sources in Owerri
metropolis, Nigeria was studied for quality. The selected water sources were
borehole, Otamiri River, Nworie Rivers, tap water and rain water. Results of bio-load
study of the water sources revealed borehole water to have the least colony forming
units per milliliter of total heterotrophic bacterial count (THBC), total coliform count
(TCC), total Salmonella-Shigella count (TSSC), and total fungal count (TFC), as against
the Otamiri River with the highest values. Physicochemical characteristics of water
sources studied were within permissible limit of World Health Organization (WHO)
standards for domestic use. The high percentage occurrence of Salmonella species
among other bacterial genera in the studied water sources raises a health concern.
These could be behind the high incidence of diarrhoea and typhoid infections,
routinely reported in the clinics within the metropolis. With these findings, there is
need for public water supply authority within Owerri metropolis to improve in quality
of water distributed. The present study has shown the recent biophysical
characteristics of domestic water sources in Owerri metropolis, Nigeria.
1066-1071 | JRB | 2013 | Vol 3 | No 6
An International
Authors:
Nwachukwu MI
1*
,
Eziuzor SC
2
, Duru MKC
3
,
Nwachukwu IO
1
,
Ukaga CN
4
, Udujih OS
1
and
Udujih GO
5
.

Institution:
1. Department of Microbiology,
Imo State University, P.M.B.
2000, Owerri, Nigeria.

2. Department of Microbiology,
Rhema University, P.M.B.
7021, Aba, Nigeria.

3. Department of Biochemistry,
Abia State University, P.M.B.
2000, Uturu, Nigeria.

4. Department of Animal and
Environmental Biological
Sciences, Imo State University,
P.M.B. 2000, Owerri, Nigeria.

5. Department of Public Health,
Federal University of
Technology, P.M.B. 1526,
Owerri, Imo State, Nigeria.

Nwachukwu MI.

Email:

Web Address:
Dates:
Received: 16 Oct 2012 Accepted: 05 Aug 2013 Published: 11 Nov 2013
Article Citation:
Nwachukwu MI, Eziuzor SC, Duru MKC, Nwachukwu IO, Ukaga CN, Udujih OS and Udujih GO.
Recent biophysical characteristics of domestic water sources in Owerri Metropolis, Nigeria.
Original Research

INTRODUCTION
Water of good quality is very important to health
and mans continued existence. The potable water
provision to rural and urban population prevents health
hazards (Lemo, 2002). Hence the principal objectives of
municipal water are the production and distribution of
safe water that is fit for human consumption (USEPA,
2001). Therefore before describing water as potable, it
has to be confirmed with certain physical, chemical and
microbiological standards which ensure that the water is
potable and safe for drinking purposes (Tebutt, 1983).
However, potable water have to be free from disease
producing microorganisms and chemical substances
deleterious to health (Ihekoronye and Ngoddy, 1985).
Water can be obtained from a number of sources
such as streams, lakes, rivers, ponds, rain, springs and
wells (Chukwura, 2001). Raymond 1992 says, Clean,
pure and safe water only exist briefly in nature and is
immediately polluted by prevailing environmental
factors and human activities. Water from most sources is
therefore unfit for immediate consumption without
treatment. The consequences of water borne bacterial
and viral infections have been well established along
with chemical contamination, which is known to cause
some deadly effect (Edema et al., 2001; Fapetu, 2000).
It is essential that water for domestic use be
examined frequently as contamination may be
intermittent. And considering the global data, morbidity
of diarrhoea disease is greater than 1.5 million and
mortality is 4 million with more than 2 billion people
being at risk. The WHO (2003) and UNICEF (2004)
have reported that 80% of sickness and death among
children in the world are caused by unsafe drinking
water. Although municipal water is distributed to large
population through closed network, but very often,
consumers are exposed to risk of water borne diseases
due to inadequate treatment of water (Antonine and
Dante, 2008; Fapetu, 2000). This study therefore is
aimed at providing recent information on the
microbiological and physiochemical characteristics of
domestic water sources in Owerri metropolis Nigeria.
This will reveal the water source or sources that could be
certified suitable for domestic usages.

Water collection
Water samples from different sources which
include borehole, Otamiri and Nworie rivers, tap water
and rainwater were collected within Owerri metropolis
and analyzed. The samples were randomly collected
from highly dependable points where residents usually
would collect their water for domestic use. Samples were
collected aseptically using sterilized 500 ml glass bottles
following the guideline of APHA (1998) and
WHO (1984) for sampling various water sources.
However, the river water sample was collected using the
method of Onyeagba et al., (2004). The collected
samples were labeled appropriately and transported to
the laboratory in an ice cool pack for analysis within
24 hours.
Bio-load study
The standard methods for the isolation and
identification of microorganisms as described by
Cappucino et al., (1992) and Onyeagba et al., (2004)
were adopted in the analyses. All the samples were
ten-fold serially diluted before being plated out using the
spread plate technique in triplicates for total
heterotrophic bacteria, count (THBC) using nutrient
agar, total coliform count (TCC) using MacConkey agar,
total Vibrio count (TVC) using thiosulphate citrate bile
salt agar, total Salmonella-Shigella count (TSSC) using
Salmonella-Shigella agar, and total fungal count (TFC)
using Sabouraud dextrose agar. All the plates were
incubated for 18 to 24 hours at 37
o
C except for
fungal count that was incubated for 3 to 5 days at
room temperature (about 26 to 32
o
C). Representative
colonies were streaked, purified, and identified
through biochemical, microscopic and macroscopic
Nwachukwu et al., 2013
observations according to Gehardt (1994) and
identification based on Holt et al., (1994).
Determination of physiochemical characteristics
Physical and chemical indices of the water
sources include colour, taste, odour, pH. Iron, total
alkalinity, chloride, biological oxygen demand (BOD),
chemical oxygen demand (COD), nitrate, conductivity,
total dissolved solids (TDS) and turbidity were
determined according to standard methods described by
APHA (1998).

RESULTS
Result of the bio-load of water sources analyzed
is shown in figure 1. The result revealed that the total
heterotrophic bacteria count (THBC) ranged between
1.5x10
2
to 1.5x10
3
cfu/ml. The total coliform count
(TCC) was in the range 1.0 to 2.0x10
2
cfu/ml, the total
Samonella/ Shigella count (TSSC) ranged from 1.5 to
2.5x10
2
cfu/ml, the total Vibrio count (TVC) ranged
from 2.5 to 7.2x10
2
cfu/ml, and total fungal count (TFC)
ranged from 2.5 to 4.0x10 cfu/ml. The findings as shown
in figure 1, make borehole water the best among the
studied water sources with no Vibrio and fungal growth;
and lowest in terms of bio-load. Otamiri River had the
highest bio-load in the present study. This makes it the
most microbiological polluted among the water sources
analyzed. Nworie River was the highest in total coliform
while tap water produced the highest value of total
fungal count. Rain water was next to borehole water in
terms of bio-load.
Statistical analysis revealed that there was
significant difference at 0.05 in the load of different
microbial groups from the different water sources
analyzed.
The overall percentage occurrence of the
different genera of bacteria and fungi isolated from the
water sources are presented in figures 2 and 3,
respectively. The bacterial percentage occurrence
revealed Salmonella (21.7%) to be highest in occurrence
as compared to the ties of Micrococcus (4.35%),
Klebsiella (4.35%) and Enterobacter (4.35%) as isolated
and analyzed. The percentage occurrence of fungi genera
isolated revealed that Aspergillus (42.85%) as the highest
and the ties of Cryptococcus (14.28%) and
Saccharomyces (14.28%) as lowest.
Statistical analysis revealed a significant
difference at 0.05 in the percentage occurrence of
bacterial and fungal isolates analyzed from the water
sources.

C
e
l
l

D
e
n
s
i
t
y

(
c
f
u
/
m
i
)

Figure 1. Bio-load of different water sources analyzed recently in
Owerri metropolis, Nigeria.
*A-borehole, B-Otamiri river, C-Nworie river, D-tap water, E-rainwater
Water Samples

The physicochemical characteristics analyzed are
shown in table 1. The water sources had pH near
neutrality in the range of 6.70 to 6.92. The borehole,
Otamiri, tap water and rainwater water sources were all
colourless. The colour and taste of borehole, Otamiri, tap
water and rainwater water sources were not
objectionable, while that of Nworie was objectionable.
The overall result showed that values for most
physicochemical indices considered in this study were
within the permissible limit as stipulated by WHO.

Figure 2. Overall percentage occurrence of different bacterial genera isolated
from water sources in Owerri metropolis, Nigeria.
Bacteria genera
0
5
10
15
20
25
P
e
r
c
e
n
t
a
g
e

o
c
c
u
r
e
n
c
e

(
%
)
Bacteria genera
Table 1. Physicochemical characteristics of water sources in Owerri metropolis
TCU-true colour unit, no-not objectionable, ob-objectionable, NTU-nephlometric turbidity units.
A-borehole, B-Otamiri, C-Nworie, D-tap water, E-rainwater
Parameters Water sources Tolerance
A B C D E WHO
Colour (TCU) ( Units) colour less colour less dull colour less colour less 500
Odour no no ob ob no no
Taste no ob ob ob No no
pH 6.7 6.92 6.86 6.92 6.82 7.0 - 8.50
Conductivity (s/cm) 146.2 23.6 45.5 28.4 3.4 500
Turbidity ( NTU) 1.0 20.37 7.77 00.0 1.5 50
Alkalinity (mg/ l) 0.0 2.00 5.00 04.0 1.0 600
Chlorine (mg/l) 0.0 0.00 0.00 00.0 0.0 200
Total Iron (mg/ l) 0.1 0.1 0.1 0.1 0.1 0.1
BOD (mg/l) 1.3 1.38 1.48 01.2 1.3 2.0
COD (mg/l) 121.45 137.18 137.18 120.2 117.58 196
TDS (mg/l) 0.2 11.7 11.7 0.1 0.1 -
DISCUSSION
The water sources in Owerri metropolis as
analyzed have shown a best option in recent times for
domestic usage. The borehole water source has the least
bio-load and chemical components thereby making it the
best source of water for domestic use among the water
sources studied. This observation could be behind the
high rate of sinking of borehole wells within Owerri
metropolis in recent times. Its low bio-load could be
attributed to the fact that it is a ground water and there is
low infiltration of pollutants from the top soil
downwards through capillary action (Chukwura, 2001;
Edema et al., 2001). Rain water which is supposed to be
the cleanest source of water by nature was the second
best in the present study. The observed low bio-load of
rain water could be due to the purification process that
takes place during condensation while its relegation to
second best could be due to incessant and reckless air
pollution from diverse sources (Nwachukwu and
otukunefor, 2006; Fapetu, 2000).
The WHO standard for domestic water supplies
which recommends a 100 cfu/ml or less for total
heterotrophic bacterial count and a zero coliform per
100ml of water was compared to results of this study
(WHO, 2003). From the observed results, only borehole
water source was acceptable while Otamiri River,
Nworie Rivers, tap water and rain water sources were
unacceptable for domestic and drinking purposes. This
study affirms a previous study, which revealed that
borehole water source has a good water acceptable
quality, microbiologically (Nwachukwu and Otokunefor,
2006).
The high percentage occurrence of Salmonella
species among other bacterial genera is a strong causal
agent. The observed high percentage occurrence of
Salmonella species in the studied water sources could be
associated to high diarrhoea and typhoid infections that
are routinely reported in the clinics within Owerri
metropolis.

CONCLUSION
Physicochemical characteristics of the water
sources in this study fall within WHO standards for
domestic use whereas the observed bio-load of the water
sources followed the order Otamiri River > Nworie River
> tap water > rain water > borehole. Borehole was the
best among the studied water sources. As inhabitants of
Owerri metropolis glamour for improvement in public
0
5
10
15
20
25
30
35
40
45
Cryptococcus sp. Candida sp. Saccharomyces sp. Aspergillus sp.
P
e
r
c
e
n
t
a
g
e

o
c
c
u
r
e
n
c
e

(
%
)
Fungal genera
Figure 3. Overall percentage occurrence of different fungal genera isolated from water sources in
Owerri metropolis, Nigeria.

water supply by public water supply authority, the
findings of the present study have also shown that the
improvement should as well include the quality of water
distributed. Efficient distribution of portable water by
public water supply authority used to be the pride of the
metropolis in the past.

REFERENCES
Antonine JPD and Dante C. 2008. Chemical levels in
drinking water. Applied Environmental Microbiology, 66
(6): 2520 2525.

American Public Health Association (APHA). 1998.
Standard Methods for the Examination of Water and
Wastewater. 20
th
ed. Washington, DC.

Chukwura EI. 2001. Aquatic Microbiology. Octoba
Press, Onitsha, Nigeria. 67 77.

Cappucinno James G and Sherman W. 1992.
Microbiology: A Laboratory Manual. 3
rd
ed. Benjamin
Cummings, California. 25 30.

Edema MO, Omemu AM and Fapetu OM. 2001.
Microbiology and physico-chemical analysis of different
sources of drinking water in Abeokuta, Nigeria. Nigerian
Journal of Microbiology, 15(1): 57 61.

Fapetu, OM. 2000. Comparative analysis of different
sources of drinking water in Abeokuta South L.G.A.,
Ogun state. BS.c thesis, University of Agriculture,
Abeokuta.

Gerhardt P. 1994. Methods for General and Molecular
Bacteriology (ed). American Society for Microbiology,
ASM Press, Washington, DC.

Holt JG, Bergey DH (ed.). 1994. Bergeys Manual of
Determinative Bacteriology, 9
th
ed. Williams and
Wilkins Co., Baltimore.

Ihekoronye AI and Ngoddy PO. 1985. Integrated Food
Sciences and Technology for the Tropics. Macmillan
Education Ltd. London and Oxford.95 195.

Lemo OO. 2002. Bacteriology Determination of Water
with Long Term Storage. BS.c thesis, University of
Agriculture, Abeokuta UNAAB, Abeokuta. 40 41.

Nwachukwu CI and Otokunefor TV. 2006.
Bacteriological quality of drinking water supplies in the
University of Port Harcourt, Nigeria. Nigerian Journal of
Microbiology, 20(3): 1383 1388.

Onyeagba A, Ugbogu OC, Kanu IJ and Ogbu O.
2004. Laboratory Guide for Microbiology. Crystal
Publishers, Owerri, Nigeria.

Raymond F. 1992. Problems of Water Supplies. EB and
Sons Ltd UK. 123 126.

Tebutt THY. 1983. Principles of Quality Control.
Pergamon publishers, England.

UNICEF. 2004. Water. Environment and Sanitation.
World Water Day 2004. Available online at
www.unicef.org//wes/index.html.

USEPA. 2001. Current Drinking Water Standards.
United States Environmental Protection Agency,
Washington, USA.

WHO. 2003. Water Supply, Sanitation and Hygiene
Development. Water. Sanitation and Health WHO,
Geneva.

WHO. 1984. Guidelines for Drinking Quality. Drinking
Water Quality Control in Small Community Supplies,
WHO, Geneva. Switzerland 3, 121-130.

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Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae and
the moth, Philosamia ricini
Keywords:
Mucopolysaccharides, Rhabdome.
ABSTRACT:

Mucopolysaccharides were detected by histochemical methods in the
crystalline cones of both the butterfly (Pieris brassicae) and the moth (Philosamia
ricini) commonly known as large cabbage white and eri silk moth respectively, but
they were absent in the rhabdome part of both the insects. The mucopolysaccharides
were extracted by biochemical method and the subsequent electrophoretic analysis
revealed that they were similar to chondroitin 4 sulfate. Moreover, chromatographic
analysis revealed different sugar components in the eyes of the two insects. It is
concluded that acid mucopolysaccharides have structural and other physiological roles
in the visual apparatus but no part in light and dark or photoperiodic adaptations.
1072-1085 | JRB | 2013 | Vol 3 | No 6
An International Scientific
Research Journal
Authors:
Bendang Ao
*
and
Sentimenla.

Institution:
Department of Zoology,
School of Sciences,
Nagaland University,
Lumami - 798627,
Nagaland, India

Bendang Ao.

Email:

Web Address:
Dates:
Received: 13 Mar 2013 Accepted: 21 Sep 2013 Published: 11 Nov 2013
Article Citation:
Bendang Ao and Sentimenla.
Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae and the moth,
Philosamia ricini.
Original Research

INTRODUCTION
Kennedy and White (1983) introduced the term
mucopolysaccharides to describe 2-amino-2-
deoxyhexose containing polysaccharides of animal origin
and occurring either as free polysaccharides or as their
protein derivative. They can be those that contain uronic
acid and those that are neutral. Acid
mucopolysaccharides (AMPs) come under the second
class. Acid mucopolysaccharides (AMPs) may be further
sulphated (SMP) or non sulphated e.g., chondroitin
sulphate and hyaluronic acid respectively. These terms
i . e. , AMPs and SMPs (sul phat ed aci d
mucopolysaccharides) appear to provide an adequate
description and also have the added advantage of
continuous use (Jaques, 1977).
Me yer ( 1938) c oi n ed t h e t er m
mucopolysaccharides to include all substances with
similar physico-chemical properties isolated from
connective tissues. Later on, the terms
gl ycosaminogl ycans gl ycopr ot ei ns and
mucoproteins were used, but they failed to distinguish
between bacterial polysaccharides and antibiotics
containing amino sugars. But these terms are still found
in literature.
Compound eyes of insects include the lens
system, a retina and underlying optic ganglia. Lens is a
modified cuticle and is composed of the cornea and
underlying crystalline cone. Immediately behind the
crystalline cone are the longitudinal sensory elements or
the retinula cells. The inner sides of the retinula cells
collectively secrete an internal light trapping rod-like
structure known as rhabdom.
Carney (1994) had indi cat ed that
glycosaminoglycans may have specific biological
functions conferred upon them because of specific
sequences within the carbohydrate chain.
Glycosaminoglycan is the systematic name for the
carbohydrate residues which form linear chains of
alternating acidic and basic monosaccharides. The basic
units are usually N-acetylated and sometimes N-sulfated,
while the acidic units are sometimes O-sulfated
(Kennedy and White, 1983).
It is to be noted that glycosaminoglycans always
come within the mucopolysaccharides category
irrespective of the ways in which the term has been used,
and it is now known that glycosaminoglycans are
attached covalently to proteins. Therefore, AMPs
actually refer to glycosaminoglycans of a proteoglycan
plus, sometimes a few amino acid units.
Presence of acid mucopolysaccharides in the
visual system of vertebrates are well documented. For
example, they have been reported in the bovine cornea
(Coster et al., 1987; Funderburgh et al., 1996; Corpuz
et al., 1996; Plaas et al., 2001; Achur et al., 2004 and
Conrad et al., 2010), in the eye of rabbit (Yue et al.,
1984; Lutjen Drecoll, 1990; Fitzsimmons et al., 1992;
Takahashi et al., 1993; Goes et al., 1999; Kato et al.,
1999), in chick cornea (Conrad et al., 1977; Li et al.,
1992; Mc Adams and McLoon 1995), in human and
rabbit cornea (Freund et al., 1995; Tai et al., 1997), in
calf lens capsule (Mohan and Spiro 1991), and in the
corneal stroma of squid (Anseth, 1961 and Moozar and
Moozar, 1973).
Other visual apparatuses where AMPs have been
reported are in the cornea of elasmobranchs (Balazs,
1965), vitreous body of the eye of squids (Balazs et al.,
1965), in aqueous and ciliary body (Cole, 1970;
Schachtschabel et al., 1977), interstitial matrix
surrounding the photoreceptor cell of the cattle (Berman
and Bach, 1968; Berman, 1969), inter photoreceptor
matrix of vertebrate (Rolich, 1970), sclera of ox (Robert
and Robert, 1967) etc. In the case of insects, AMPs have
also been reported in the compounds eyes of Periplaneta
americana, Belostoma sp (Dey, 1976), Palaemon sp,
Limulus polyphemus and Macrobrachium birmanicum
(Dey et al., 1978), Musca domestica, Apis cerana
indica (Dey, 1980)

Bendang, 2013
Acid mucopolysaccharides play several
important physiological roles owing to their capacity to
bind and hold water (Ogston, 1970; Ogston and Wells,
1972; Wells, 1973b). They serve as natural lubricants in
the joints, impart elasticity to connective tissue, and are a
component of cartilage and ligaments. They are also
involved in support and motor functions, and also have
bactericidal properties. It is also known that many
diseases such as collagenosis, mucoplysaccharidosis, and
rheumatism etc which are correlated with aging, are also
a result of disorders in mucopolysaccharides metabolism
which lead to compositional changes of connective tissue
and of the body fluids.
With this view a study was done in the
compound eye of the insects viz., butterfly,
Pieris brassicae and moth, Philosamia ricini with
regards to the occurrence of acid mucopolysaccharides,
and their possible functions in the eyes have been
discussed.

The eyes were separated from live insects and
fixed in 10% buffered formalin until they were used.
Histochemical study:
The tissues were embedded in paraffin and 8
thick sections were cut by microtome. The section were
stained with Toluidine blue and Alcian blue (Humason,
1971) for detection of mucopolysaccharides.

Biochemical study according to Dietrich et al., (1977).
Extraction:
Fresh eyes (1gm) were defatted in cold acetone
for three hours and dried. The tissues were then
homogenized and suspended in 20 ml of 0.05M Tris-HCl
buffer (pH 8). To the mixture, 10 mg of trypsin was
added and then a few drops of toluene were added
forming a layer at the surface, and incubated at 37C for
24 hours. After incubation, pH of the mixture was
brought to 11 with Conc. NaOH and maintained for six
hours at room temperature. Then the pH was brought to 6
by the addition of HCl and the mixture was centrifuged
for 15 minutes at 3000rpm. To the supernatant, 0.1 ml of
2M NaCl and two volumes of ethanol were added and
kept overnight at 5C. The mixture was centrifuged for
15 minutes at 3000 rpm and the precipitate was collected
and dried. The resultant powder was re-suspended in 1
ml of 0.05M sodium acetate (pH 6.5) along with 1 mg of
DNAase and RNAase. The solution was again incubated
for 24 hours at 37C with a layer of toluene. After
incubation, 0.1 ml of 2M Nacl and two volumes of
ethanol were added to the solution and kept overnight at
5C. It was then centrifuged for fifteen minutes at 3000
rpm and precipitate was collected and dried. The
resultant powder was dissolved in 0.5 ml of water, heated
at 100C for two minutes and analyzed by paper
chromatography and electrophoresis.
Chromatography:
The extract was hydrolyzed with 6N HCl at
100C for 12 hours. The acid hydrolysate was then
evaporated to dryness. The dried residue was then
dissolved in 0.5 ml of distilled water and spotted in
whatman No 1 filter paper and ascending paper
chromatograms were run using butanol, acetic acid and
water in the ratio of 4:1:1 (v/v) as solvent (Giri and
Nigam, 1954).
The chromatogram was developed with silver-
nitrate (0.1 ml of saturated solution in 20 ml of acetone)
and sodium hydroxide (0.5 gm of NaOH in 25 ml of
rectified spirit) as suggested by Trevelyan et al., (1950).
The chromatogram was then washed in 6N ammonium
hydroxide for 10 minutes and then washed in running
water and dried at room temperature.
Electrophoresis:
This was according to the method as described
by Leitner and Kerby, (1954). Streaks of the acid
mucopolysaccharide samples were applied on Whatman
No.1 paper strips using 0.1M phosphate buffer (pH 6.6)
at 4v/cm for 8 hours. After removal from the
Bendang, 2013

electrophorectic apparatus, the paper strips were dried at
room temperature and stained with Toluidine blue
(0.04% in 80% acetone). The staining of the strips was
followed by 2-3 rinsing in 0.1% acetic acid and then 2-3
times in H
2
O. The strips were then dried at room
temperature.
OBSERVATIONS
Histochemical observations:
Lens cuticle of the butterfly, Pieris brassicae:
When the sections of the eyes were stained with
toluidine blue, the cornea and crystalline cone became
purple in color showing metachromasia (Photoplate 1)
i . e. , i ndi cat i ng t he pr esence of aci d
mucopolysaccharides, while the region of the rhabdom
was orthochromatic (blue in colour) and therefore
devoid of acid mucopolysaccharides. Similarly, when the
eyes were stained with alcian blue, the lens and
crystalline cone became purple in colour (Photoplate 2)
whi ch i ndi cat es t he pr esence of aci d
mucopolysaccharides. (Fig 1)
Lens cuticle of the moth, Philosamia ricini:
When the sections were stained with toluidine
blue, the cornea as well as crystalline cone became
purple in colour (Photoplate 3) showing the presence of
mucopolysaccharides. The more intense reactions were
observed towards the corneal lens. The rhabdom region
however gave a blue colour reaction i.e. the region is
orthochromatic (Photoplate 4). When the eyes were
stained with alcain blue the corneal lens and crystalline
cone became purple in colour indicating the presence of
AMPs, while the rhabdom became blue in colour which
indicates absence of AMPs. (Fig 2)
Biochemical observations:
Chromatographic analysis of the acid
mucopolysaccharides extract showed the presence of
three sugars viz lactose, galactose and xylose in case of
Pieris brassicae and galactose, xylose and rhamnose in
the case of Philosamia ricini (Figure 3 and 4;
Table 1 and 2).
Electrophorectic movement pattern of the crude
extracts of the acid mucopolysaccharides from the eyes
of Pieris brassicae and Philosamia ricini, when
compar ed wi t h s ever al st andar d a ci d
mucop ol ys a cch a r i des sh owed t h a t t h e
mucopolysaccharides extracted resemble chondroitin
4-sulfate (Figure 5 and 6; Table 3 and 4).

DISCUSSION
Several workers like Miao et al., (1996), Groves
et al., (2005), Manton et al., (2007), Fthenou et al.,
(2006, 2008) etc. have studied the influence of
glycosaminoglycans on cell division, differentiation,
responses to growth factors, adhesion, migration,
peripheral nerve extension or regeneration and signal
transduction. In this regard, Bulow and Hobert, (2006)
are of the opinion that the correct development of a
multicellular organism is via a specific code contributed
by the glycosaminoglycans.
In the case of the visual apparatus, they play a
central role in the physiological maintenance of
trabecular meshwork in the eyes (Yue et al., 1984 and
Cavallotti et al., 2004). They may also have a role in
influencing keratocytes and nerve growth in corneal
stroma because of their ability to bind together (Cornard
et al., 2010). They, and their core proteins also have
important physiological and homeostatic roles e.g.
during inflammation and immune response (Park et al.,
2001; Li et al., 2002; Wang et al., 2005).
AMPs influence tissue osmotic pressure not only
by influencing the water balance, but also by introducing
excess swelling pressure which is balanced by an internal
structural resistance (Ogston, 1970). Moreover, AMPs
play important roles in water binding and maintenance
of tissue osmotic pressure (Ogston and Wells, 1972).
Payrau et al., (1967) observed that the transparency of
the cornea is based on the state of hydration of tissue.
They based this on the fact that the corneal stroma of
most vertebrates, including mammals, birds and teleosts
Bendang, 2013
absorb water wherever free water is accessible. In
contrast, according to Maurice and Riley (1970) odema
of the cornea leads to disorganization of its structure and
less transparency, but dehydration does not appear to
have serious optical affects. Maurice (1972) suggested
that the presence of AMPs in the cornea is mainly
responsible for the dehydration properties of the tissue
and hence transparency. This is supported by workers
like Hedbys (1961, 1963); Kikkawa and Hirayama
(1970); Bettelheim and Plessy (1975); Lee and Wilson
(1981) and Castoro et al., (1988).
AMPs have also been suggested to play a major
role in the structural organization of intracellular matrix
via electrostatic and steric interactions with other
macromolecules of the matrix, such as collagen and
elastin (Kobayashi and Pedrini, 1973). Similarly, Ogston
and Wells, (1972) have suggested that AMPs help in the
maintenance of mechanical flexibility and elasticity of
tissues. Ogston, (1966a) and Katchalsky, (1964) have
shown that acid mucopolysaccharides possess high water
binding capacities.
Multiple types of chondroitin sulphate
proteoglycans are seen in vertebrates and they greatly
influence development and tissue mechanics. For
example, the chondroitin chains in the nematode
Caenorhabditis elegans are not sulphated, but are
nevertheless essential for embryonic development and
vulval morphogenesis (Olson et al., 2006). Chondroitin
and dermatan proteoglycans have also been the subject
of much interest as inhibitors of axon growth and have
been shown to be important components of the glial scar
that prevents axon regeneration (Rhodes and Fawcett,
2004).
The role of mucopolysaccharides in
pathogenicity has been widely reviewed. For instance,
they are responsible for calcification of bones (Rubin and
Howard, 1950), dermal thickening in acromegalic
patients (Matsuoka et al., 1982), involved in inborn
Bendang 2013
Fig 1. Histochemical observations of Lens cuticle of the
butterfly, Pieris brassicae
Fig 2. Histochemical observations of Lens cuticle of the
moth, Philosamia ricini

errors of metabolism and/ or storage disorders (Matalon
et al., 1974a; Hall et al., 1978; Neufeld and Fratantoni,
1970; McKusick et al., 1978), maintenance of retinal
structure and neural tube closure in Knobloch syndrome
(Sertie et al., 2000) and treatment of diabetic
nephropathy (Gambaro and Van Der Woude, 2000).
Matthews (1959) and Oosawa (1971) have
suggested that one of the characteristic properties of
mucopolysaccharides is the selective association or
binding with small inorganic cations, especially H
+
, Na
+
,
and Ca
++
, and also with cationic groups of
macromolecules. In these regard, Farber and Schubert
(1957) and Urist et al., (1968) have also found a small
preference for binding Ca
++
over Na
+
in chondroitin
sulphate. Matthews (1975) thus suggested that these
substances act as a store for Ca
++
in cartilage tissue and
that is the reason for their specific roles in tissue-
calcification. Some roles of AMPs, especially in
arthropodan cuticle have been reported by Meenakshi
and Scheer (1959) and Sundara Rajulu (1969) in terms of
calcification of the cuticle of Hemigrapsus nudus and
Cingalobolus bugnioni respectively. Krishnan (1965) has
suggested that AMPs may be associated with -S-S-
bonding of the cuticle in the scorpion Palaemonetes
swammerdami.
Si n c e t h e o c c u r r e n c e o f a c i d
mucopolysaccharides is not a general feature of the
arthropod cuticle and it occurs in some special types of
cuticle where it performs some special functions
(Meenakshi and Scheer, 1959; Sundara Rajulu, 1969;
Krishnan, 1965 and Raghuvarman et al., 1998), it is
reasonable to presume that the specific occurrence of
mucopolysaccharides in the lens cuticle and the
crystalline cone may have a bearing on the visual system
of the insects. Keeping the above account in view it is
possible to assume a role of AMPs in the lens-cuticle of
insects.
The lens-cuticle as already stated, besides
playing a general defensive role, performs a special
optical function of conducting light rays to the inner
rhabdomere. It is possible to presume that the
transparency of the lens-cuticle, which is more than that
of other types of cuticle (e.g. body cuticle), may be
affected by the occurrence of mucopolysaccharides
(Anseth and Fransson, 1970). Similarly, Freund et al.,
(1995) also reported that the presence of AMPs in human
and rabbit cornea is related to transparency. It is known
that the bulk of cornea of vertebrate eye is the stroma,
which functions as a supporting structure and is adapted
for the transmission of a high percentage of incident light
of visible-wave length (Maurice, 1969). Anseth and
Fransson (1970) have found that during chick corneal
development, the occurrence of a highly sulfated keratan
sulfate is associated with rise in the transparency of
stroma. They have also suggested that stromal
transparency is correlated with the presence of normal
Bendang 2013
Table 2: Ascending Paper chromatogram of some
standard sugar components. (Solvent used is butanol,
acetic acid and water in the ratio of 4: 1:1 v/v)
Sugar Rf value
Raffinose 0.03
Lactose 0.05
Glucose 0.10
Sucrose 0.13
Galactose 0.18
Mannose 0.25
Fructose 0.28
Xylose 0.34
Ribose 0.38
Table 1: Ascending paper chromatogram of sugar
components of the butterfly, Pieris brassicae and the
moth, Philosamia ricini. (Solvent used is butanol,
acetic acid and water in the ratio of 4: 1:1 v/v)
Insect Rf value Identification
Butterfly,
Pieris brassicae
0.05 Lactose
0.18 Galactose
0.33 Xylose

Moth,
Philosamia ricini
0.16 Galactose
0.33 Xylose
0.43 Rhamnose
proportions of keratan sulfate and chondroitin 4-sulfate.
Funderburgh et al., (1996) have reported that
keratan proteoglycans are the major proteoglycans of the
bovine cornea and secreted by keratocytes in the corneal
stroma and they are thought to play an important role in
corneal structure and physiology, particularly in the
maintenance of corneal transparency. Blochberger et al.,
(1992), has reported that corneal keratan sulfate
proteoglycans contribute to corneal transparency in
chick. Takahashi et al., (1993) have also reported that
keratan sulfate and dermatan sulfate proteoglycans are
associated with collagen in foetal rabbit cornea.
Transparency of the corneal stroma depends partially on
the degree of spatial order of its collagen fibrils which
are narrow in diameter and closely packed in a regular
array (Maurice, 1957; Cox et al., 1970; Benedek, 1971;
Mc Cally and Farrell, 1990 and Bron, 2001). Mc Adams
and Mc Loon (1995) have shown that retinal axons grow
in the presence of chondroitin sulphate and keratan
sulfate proteoglycans and that these proteoglycans helps
in developing chick visual pathway.
Many studies that focused on corneal swelling
behavior have noted a gradual decrease in swelling from
the posterior to anterior side (Van Horn et al., 1975;
Bendang 2013
Fig. 6: Paper electrophorectic movement pattern of
the crude mucopolysaccharides from the eyes of the
moth Philosamia ricini
Fig. 5: Paper electrophorectic movement patterns
of the crude mucopolysaccharides from the eyes of
the butterfly, Pieris brassicae.
Fig. 3: Ascending paper chromatogram showing the
sugar components of the mucopolysaccharides from
the eye of the butterfly, Pieris brassicae.
Fig. 4: Ascending paper chromatogram showing the
sugar components of the mucopolysaccharides from
the eye of the moth Philosamia ricini.
Bettelheim and Plessy 1975; Castoro et al., 1988 and
Cristol et al., 1992) and this was thought to be related to
the organization of the collagen lamellae and the
presence of different types of proteoglycans. In the
posterior part, keratan sulfate, a more hydrophilic
proteoglycan is prevalent, whereas in the anterior part
dermatan sulfate, a much less hydrophilic proteoglycan,
is present (Bettelheim and Plessy 1975; Castoro et al.
1988). An interesting conclusion was drawn by Muller et
al., (2001) while studying the differential behaviour of
the anterior and posterior stroma during corneal swelling,
that it is the high negative charge of the
glycosaminoglycan components of the proteoglycans that
is responsible for the corneal swelling due to electrostatic
repulsion between acidic groups. They also suggested
that the structural stability of the anterior stroma under
condition of extreme hydration imply an important role
for this zone in the maintenance of corneal curvature and
that this stability is determined by the tight interweave of
the stromal lamellae.
It is now known that the pH value is a decisive
factor for the taking of water by the cornea (Cejkova and
Brettschneider, 1969). The protein polysaccharide
complex provides a more stable and specific
configuration within the molecules than electro-static
linkage could. For the cornea to remain transparent, it is
essential that an active mechanism counter the natural
tendency of the stroma to increase its hydration, swelling
and opacity. It may be noted here that the non - swelling
properties of elasmobranch cornea is supposed to be due
to the high mannose content in their structural proteins
(Moozar and Moozar, 1972).
It is well-established that one of the corneal
limiting cell layers i.e., the corneal endothelium,
transports fluid at a substantial rate and that this transport
is essential to maintain normal stromal hydration
(Maurice, 1972; Candia, 1976; Candia and Zamudio,
1995; Narula et al., 1992; Bonanno et al., 1989 and Yang
et al., 2000). Anseth and Fransson, (1969) had
demonstrated the synthesis of AMPs by corneal
epithelial and stromal cells, and that they are important
in maintaining the corneal structure in relation to its
environment. Deb and Raghuvarman (1994) have also
observed that glycosaminoglycans are essential for the
maintenance of corneal structure and function.
Acid mucopolysaccharides thus detected in the
compound eyes of the butterfly, pieris brassicae and the
moth, Philosamia ricini may play an important role in
visual excitation, when light rays pass through the outer
epicuticle to the inner endocuticular region (crystalline
cone) - the sites of AMPs, due to the fact that they act as
a selective ion barrier (Jeanloz, 1970). It may also be
noted that they are present not only in the corneal lens
but also in the crystalline cone, which are in close
connection with the inner rhabdomeres (the actual sites
of photochemical reactions), the products of which may
depolarize the membrane of the retinula cells and initiate
impulse formation (Wigglesworth, 1965). Further,
mucopolysaccharides may play a role in increasing
transparency of lens-cuticle. In this context, it is worth
mentioning that during corneal development of
Bendang, 2013
Table 3: Paper electrophorectic movement patterns of
the crude mucopolysaccharides from the eyes of the
butterfly, Pieris brassicae and the moth, Philosamia
ricini. (Solvent used is phosphate buffer of pH 6.5)
Insect
Distancetravelled
(cms)
Acid mucopolysaccharide
type
Butterfly,
Pieris brassicae
6.4 Chondroitin 4-sulfate
Moth,
Philosamia ricini
6.8 Chondroitin 4-sulfate
Table 4: Paper electrophorectic movement patterns of
some standard mucopolysaccharides. (Solvent used is
phosphate buffer of pH 6.5)
Standard
mucopolysaccharides
Distance travelled
(cms)
Heparin 5.5
Chondroitin 4-sulfate 6.6
Heparan sulfate 7.2
Chondroitin 6-sulfate 7.6
Keratan sulfate 8.7
Dermatan sulfate 10.0
vertebrates, rise in transparency of stroma was found to
be associated with occurrence of mucopolysaccharides
(Anseth and Fransson, 1970).
It is thus concluded that AMPs do indeed play
various roles in the physiology of vision, but no
photoperiodic adaptational mechanisms can be attributed
to them.

CONCLUSIONS
The present investigation revealed that
mucopolysaccharides are present in the ocular tissues
(crystalline cones, but absent in the rhabdome) of both
the insects studied i.e., Pieris brassicae, and
Philosamia ricini. Moreover, the analysis of sugar
components show that the ocular tissues of both the
insects have similar sugars galactose and xylose,
except for two different sugar components i.e., lactose
(in Pieris brassicae) and rhamnose (in Philosamia
ricini), but no definitive conclusion can be drawn on the
matter of this difference pending further studies. It is
thus concluded that acid mucopolysaccharides have
structural and other physiological roles in the visual
apparatus but no part in light and dark or photoperiodic
adaptations.

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the reader to make a reliable decision on probable interest. Do not use all caps; instead use caps only at the first word of the title and/or at
scientific names, abbreviations etc., Center the authors' initials and last names directly below the title.

Abstract

The abstract should include a hypothesis or rationale for the work, a brief description of the methods, a summary of the results, and a conclusion:
The abstract should be less than 250 words. Do not include literature citations or references to tables, figures or equations.

Keywords

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content of your article. Note that words in the title are not searchable as keywords unless they are also included in the keyword list.

Body of the Article

The introductory section of the text should include a brief statement of why the research was conducted. It should also define the
problem and present objectives along with a plan of development of the subject matter. The introductory section also usually includes a brief
survey of the relevant literature on the topic.

Materials and Methods

Provide sufficient detail so that the work may be repeated. Do not give details of methods described in readily available sources.
Instead, refer to the source and describe any modification. Figures that illustrate test apparatus and tables of treatment parameters or equipment
specifications are appropriate here.

Results and Discussion

This section describes the solution to the problem stated in the introductory section. Use figures and tables to visually supplement the
presentation of your results. The text must refer explicitly to all visuals, and you must interpret the visual elements to emphasize the evidence on
which your conclusions are based. Do not omit important negative results.
In addition, relate your findings to previous findings by identifying how and why there are differences and where there is agreement. Speculation
is encouraged, but it must be identified.

Conclusion

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suggestions for future research. The conclusion may be a subsection of the Results and Discussion section, or it may be a separate section. Data
or statements cited in your conclusion must have been stated previously in the article. Do not introduce new information in t he conclusion.

Acknowledgement

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financing. The acknowledgements will appear at the end of the text and should be limited to one or two sentences.

References

All sources cited in the text must be listed in the References, and all documents listed in the References must be cited in t he text.
Accuracy of citation is the author's responsibility.

Reference Style

References should be cited in the text in the form (Author et al, 1987) and listed in alphabetical order at the end of the article as follows:

Schernewski G, Neumann T. The trophic state of the Baltic Sea a century ago: a model simulation study. J Mar Sys., 2005;53:109
124.

Kaufman PD, Cseke LJ, Warber S, Duke JA and Brielman HL. Natural Products from plants. CRC press, Bocaralon, Florida. 1999;
15-16.

Kala CP. Ecology and Conservation of alphine meadows in the valley of flowers national park, Garhwal Himalaya. Ph.D Thesis,
Dehradun: Forest Research Institute, 1998; 75-76.

http://www.ethnobiomed.com/content/pdf/1746-4269-1-11.pdf.

Appendix

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