Journal of Research in Biology Volume 3 Issue 6

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Laboratorio de ecología y conservación de fauna Silvestre,

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

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Universidade Federal de São João del-Rei, Brazil.

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BCKV (Agri University), West Bengal, INDIA.

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School of Chemical & Biotechnology, Sastra University, Tamilnadu, INDIA.

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Jawaharlal Technological University, Hyderabad.

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University of Kordofan, Elobeid-SUDAN.

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University of Mosul, Mosul,Iraq.

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

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Hem. North Gujarat University, Patan.

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

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Federal University of Rondônia, Brazil.

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School of Life Sciences, Bharathidasan University, India.

Dr. Mrs Kavita Sharma [Botany]

Arts and commerce girl’s college Raipur (C.G.), India.

Suwattana Pruksasri [Enzyme technology, Biochemical Engineering]

Silpakorn University, Thailand.

Dr.Vishwas Balasaheb Sakhare [Reservoir Fisheries]

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.

Dr. Amit Bandyopadhyay [Sports & Exercise Physiology]

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 Biotecnología de las Plantas, Universidad Central de Las Villas

Eudriano Costa [Aquatic Bioecology]

IOUSP - Instituto Oceanográfico da Universidade de São 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.

Hérida 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.

María de la Concepción García 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 jaén, Spain.

Dr. Milan S. Stanković [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

http://jresearchbiology.com/documents/RA0363.pdf




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

Jou

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esearch

in

Biology

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

Journal of Research in Biology 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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Dayananda, 2013


Fig. 1. Percent composition of avian families represented by species

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Ahmadpour and Younes Yaghobzadeh. 2011. A three

years study of the diversity and density of waterfowl and

waders in Sorkhrud International Wetland (October 2007

–March 2010). Scientific Research and Essays,

6(30):6317-6324.

Nanda KNV, Sailaja K, Nagarjuna A. 2010. Avian

biodiversity indices and comparative chronobiology of

Uppalapadu and Nelapattu bird protected areas of

Andhra Pradesh, India. J. Zool., 5(3):148-152.

Rajashekara S and Venkatesha MG. 2011.

Community composition of aquatic birds in lakes of

Bangalore, India. J. of Env. Biol., 32(1):77-83.

Rajpar MN and Zakaria M. 2010. Density and

diversity of water bird and terrestrial bird at Paya Indah

wetland reserve, Selangor Peninsular Malaysia. J. Biol.

Sci., 10(7):658-666.

Sonobe K and Usui S. 1993. A Field Guide to the

Water Birds of Asia. Wild Bird Society of Japan, Tokyo.

William J Sutherland. 1997. Ecological Census

Techniques a handbook. Cambridge University Press

U.K.

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. Journal of Research in Biology (2013) 3(6): 1060-1065

Jou

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esearch

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Biology

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



www.jresearchbiology.com Journal of Research in Biology

An International


Authors:

Afamdi Anene1, Olivia C.

Afam-Anene2, Kelechi Ike1

and Nnamdi H. Ekekwe1

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:


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 4th 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


http://www.feedipedia.org/node/16357


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 35oC 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 Duncan’s 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 58.730±0.34a 12.79±0.22a

Pig dung 08.180±0.22d 02.97±0.17d

Cattle dung 12.920±0.16c 04.18±0.52c

Cattle Blood 21.770±0.31b 07.79±0.41b

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, Heuzé 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% (Heuzé

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). Heuzé 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 56.25±0.21a 55.54± 0.15a 56.00a± 0.00 57.42a±0.00

Crude fibre 00.32±0.08a 00.26±0.05ab 00.21± 0.01b 00.29±0.06ab

Ash 10.80±0.17b 11.09±0.15a 10.90± 0.12b 11.20±0.11a

Ether Extract 22.32±0.09a 22.64±0.07a 22.66± 0.21a 21.06±0.19a

Nitrogen Free Extract 00.17±0.04a 00.61±0.07a 00.07± 0.01a 00.16±0.07a

Moisture content 10.12±0.11b 09.84±0.12b 10.14± 0.21a 09.86±0.16b

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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Adeniji AA. 2007. Effects of replacing groundnut cake

with maggot meal in the diet of broilers. Int. J. Poult.

Sci., 6 (11): 822-825.

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larvae and pupal in three substrates of poultry droppings.

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Ibadan, Nigeria.

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domestica) meal generated from mixture of cattle blood

and wheat bran. Livestock Research for Rural

Development. 20 (12):1-5.

Aniebo AO and Owen OJ. 2010. Effects of Age and

Method of Drying on the Proximate Composition of

Housefly Larvae (Musca domestica Linnaeus) Meal

(HFLM). Pakistan Journal of Nutrition 9 (5): 485-487,

2010.

AOAC. 1995. Official Methods of analysis AOAC,

International, P. Cunniff Sixteenth edition, Vol. II

Chapter 49, 1-49. Arlington, Virginia, United States.

Awoniyi TAM and Aletor VA. 2002. Proc. 29th Ann.

Cont. Nig. Soc. of Anim. Prod. NSAP. March 17-21

2002. Fed- University of Tech. Akure, Nigeria. 170-173.

Calvert CC, Martins RD and Eby HJ. 1971. Housefly

pupae as food for poultry. Journal of Economic

Entomology. 62(1): 939.

Cao JunMing, Yan Jing, Huang YanHua, Wang

GuoXia, Zhang RongBin, Chen XiaoYing, Wen

YuanHong, Zhou TingTing. 2012. Effects of

replacement of fish meal with housefly maggot meal on

growth performance, antioxidant and non-specific

immune indexes of juvenile Litopenaeus vannamei.

J. Fish. China, 36 (4): 529-537.

Duncan DB. 1955. Multiple range and multiple F tests.

Biometrics, 11: 1-42.

Ekoue SE and Hadzi YA. 2000. Maggots production as

proteins source for young poultry in Togo - Preliminary

observations. Tropicultura, 18 (4): 212–214.

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.

Heuzé V and Tran G. 2013. Housefly maggot meal.

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and FAO. http://www.feedipedia.org/node/671.

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http://www.feedipedia.org/user/3

http://www.feedipedia.org/user/4


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

Chicken Feces as Food Source for Poultry. J. Poult. Sci.,

36 (3): 174-180.

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

sources of dietary protein in larval Heterobranchus

longifilis (Valenciennes, 1840). J. Anim. Plant Sci., 15

(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, Čičková H, Kozánek M, Martínez-Sánchez

A, Takáč 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): 587–596.

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.

Sogbesan AO, Ajuonu N, Musa BO, Adewole AM.

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

two-stage composting of pig manure. Biores. Technol.,

116: 485–491.

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factors influencing the Musca domestica L. fecundity.

MedskayaParazitol. 58: 27–30 [in Russian].

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Biology

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 MI1*,

Eziuzor SC2, Duru MKC3,

Nwachukwu IO1,

Ukaga CN4, Udujih OS1 and

Udujih GO5.

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:


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.

Journal of Research in Biology (2013) 3(6): 1066-1071

Journal of Research in Biology

Original Research

An International Scientific Research Journal

INTRODUCTION

Water of good quality is very important to health

and man’s 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 37oC except for

fungal count that was incubated for 3 to 5 days at

room temperature (about 26 to 32oC). 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

1.5x102 to 1.5x103 cfu/ml. The total coliform count

(TCC) was in the range 1.0 to 2.0x102 cfu/ml, the total

Samonella/ Shigella count (TSSC) ranged from 1.5 to

2.5x102 cfu/ml, the total Vibrio count (TVC) ranged

from 2.5 to 7.2x102 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.



Cell

Den

sity

(cfu

/mi)

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

25P

erc

en

tage

occ

ure

nce

(%

)

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 (µs/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.

Pe

rce

nta

ge o

ccu

ren

ce (

%)

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. 20th 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. 3rd 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. Bergey’s Manual of

Determinative Bacteriology, 9th 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.




Advantages


[email protected]


http://www.unicef.org/wes/index.html

Jou

rn

al of R

esearch

in

Biology

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:


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. Journal of Research in Biology (2013) 3(6): 1072-1085


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 (sulphated acid

mucopolysaccharides) appear to provide an adequate

description and also have the added advantage of

continuous use (Jaques, 1977).

M e ye r ( 1 9 3 8 ) c o i n ed t h e t e r m

“mucopolysaccharides” to include all substances with

similar physico-chemical properties isolated from

connective tissues. Later on, the terms

“glycosaminoglycans” “glycoproteins” 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 indicated 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 µ

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 37°C 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 5°C. 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 37°C 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

5°C. 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 100°C for two minutes and analyzed by paper

chromatography and electrophoresis.

Chromatography:

The extract was hydrolyzed with 6N HCl at

100°C 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 H2O. 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 . , in di ca t in g th e pr esen ce of a cid

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)

which indi cat es the presence of a cid

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

com par ed wi th s eve r a l s t an da r d a ci d

m u cop o l ys a cch a r i d e s sh o wed 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.

S i 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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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.

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Journal of Research in Biology Volume 3 Issue 6