Journal of Research in Biology Volume 3 Issue 6 by Journal of Research in Biology

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Dr. Afreenish Hassan [Microbiology] Department of Pathology, Army Medical College, Rawalpindi, Pakistan. Gurjit Singh [Soil Science] Krishi Vigyan Kendra, Amritsar, Punjab, India. Dr. Marcela Pagano [Mycology] Universidade Federal de São João del-Rei, Brazil. Dr.Amit Baran Sharangi [Horticulture] BCKV (Agri University), West Bengal, INDIA. Dr. Bhargava [Melittopalynology] School of Chemical & Biotechnology, Sastra University, Tamilnadu, INDIA. Dr. Sri Lakshmi Sunitha Merla [Plant Biotechnology] Jawaharlal Technological University, Hyderabad. Dr. Mrs. Kaiser Jamil [Biotechnology] Bhagwan Mahavir Medical Research Centre, Hyderabad, India. Ahmed Mohammed El Naim [Agronomy] University of Kordofan, Elobeid-SUDAN. Dr. Zohair Rahemo [Parasitology] University of Mosul, Mosul,Iraq. Dr. Birendra Kumar [Breeding and Genetic improvement] Central Institute of Medicinal and Aromatic Plants, Lucknow, India. Dr. Sanjay M. Dave [Ornithology and Ecology] Hem. North Gujarat University, Patan. Dr. Nand Lal [Micropropagation Technology Development] C.S.J.M. University, India. Fábio M. da Costa [Biotechnology: Integrated pest control, genetics] Federal University of Rondônia, Brazil. Marcel Avramiuc [Biologist] Stefan cel Mare University of Suceava, Romania. Dr. Meera Srivastava [Hematology , Entomology] Govt. Dungar College, Bikaner. P. Gurusaravanan [Plant Biology ,Plant Biotechnology and Plant Science] 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.

CHANDRAMOHAN [Biochemist] College of Applied Medical Sciences, King Saud University.

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

B.C. Behera [Natural product and their Bioprospecting] Agharkar Research Institute, Pune, INDIA.

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

Kuvalekar Aniket Arun [Biotechnology] Lecturer, Pune.

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

Mohd. Kamil Usmani [Entomology, Insect taxonomy] Aligarh Muslim university, Aligarh, india.

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

Dr. Lachhman Das Singla [Veterinary Parasitology] Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, India.

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

Vaclav Vetvicka [Immunomodulators and Breast Cancer] University of Louisville, Kentucky.

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

José F. González-Maya [Conservation Biology] Laboratorio de ecología y conservación de fauna Silvestre, Instituto de Ecología, UNAM, México.

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.

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

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

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

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

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

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

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

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

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

RA0363

Title of the article

Diversity of Wetland dependent birds around the Bhadra Reservoir

Page No

1054-1059

Project (BRP) area, Karnataka. Dayananda GY.

RA0374

Preliminary investigations on quantity and proximate quality of

1060-1065

maggots produced from four different sources of livestock wastes. Afamdi Anene, Olivia C. Afam-Anene, Kelechi Ike and Nnamdi H. Ekekwe.

RA0297

Recent biophysical characteristics of domestic water sources in Owerri

1066-1071

Metropolis, Nigeria. Nwachukwu MI, Eziuzor SC, Duru MKC, Nwachukwu IO, Ukaga CN, Udujih OS and Udujih GO. 4

RA0340

Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae and the moth, Philosamia ricini. Bendang Ao and Sentimenla.

1072-1085

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

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

ABSTRACT:

Institution: Department of P.G. Studies and Research in Applied Zoology, Bioscience Complex, Jnana Sahyadri, Kuvempu University, Shankaraghatta â&#x20AC;&#x201C; 577 451. Shimoga.

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.

Corresponding author: Dayananda GY.

Keywords: Wetland birds, diversity, wetlands, Bhadra Reservoir Project .

Email Id:

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

Web Address:

Dates: Received: 06 July 2013

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

Accepted: 22 July 2013

Published: 04 Sep 2013

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

Journal of Research in Biology An International Scientific Research Journal

1054-1059 | JRB | 2013 | Vol 3 | No 6

www.jresearchbiology.com

Dayananda, 2013 INTRODUCTION

percentage of birds occurrence was calculated to

Wetlands are the treasures of avifaunal species

determine the status. The modified score classes used on

richness and these are the important ecological

the basis of total bird recorded during study period i.e.,

significance areas, which serves as a major link between

1-5%= rare (R), 6-10%=Uncommon (UC), 11-13%=

the natural resources and agricultural practices. Wetlands

common (C) and >14% = Verycommon (VC) as

of lentic group form a favorable habitat to various groups

accomplished by Mc Kinnon and Philips (1993).

of animals especially waterfowl, that need food, water for drinking, wallowing and abode. Wetlands are known

RESULTS AND DISCUSSION

to be most productive and diverse ecosystems on the

A total of 68 species of birds were found

earth. Water birds are perhaps the most visible

associated with the Bhadra Reservoir. Of which 40

manifestation of faunal diversity but many other groups

species are resident, 21 residents with local migratory

also inhabit these wetlands. Wetlands are fragile

and seven are migratory. Some of the migratory birds

ecosystems, which are fast deteriorating and shrinking

recorded includes White-necked Stork, Shoveler, Pintail,

due to man made activities. India has 65,000 wetlands

Grey Plover, Curlew, Ringtailed-fishing Eagle and Black

covering an area of 4.5 million hectares (Anon, 1990).

-winged Stilt. These are winter migrants used the

The diversity of water birds obviously indicate the

wetlands for foraging, resting and other activities till

quality and healthy condition of the ecosystem in the

favorable condition of their native and some residential

country. Concerning the realm of this study, some other

wetland birds such as the herons, egrets, ibises and storks

works have been carried out by Dayananda (2009);

used the trees and shurbs as roosting site. These species

Nanda et al., (2010); Rajpar and Zakaria (2010); Mohsen

were found during the study period on the ground

et al., (2011). The aim of this study is to assess the

feeding of fishes, amphibians and crutaceans. The report

diversity of wetland birds in and around Bhadra

suggested that the wetlands are important places for

Reservoir Project area.

foraging of wetland birds. This observation got support from earlier publications which reported that, habitat has

MATERIALS AND METHODS

long been used as a predictor of bird species abundance

The checklist of wetland birds around the BRP

and variety of birds has developed different preferences

area was made by sighting the birds with 8 x 50

for habitat (Huston, 1994; Lameed, 2011). During study

binoculars. The field guides (Ali, 1996; Sonobe and

68 bird species belonging to 19 families and six orders

Usui, 1993; Inskipp and Inskipp, 1991; Fleming et al.,

were found on the wetland (Table-1). The status based

2000; Kazmierczak and Perlo, 2000; Grimmett et al.,

upon percent occurrence of bird species representing

2001) were used for bird identification. The wetland bird

different families with respect to total bird species

census was conducted in morning hours from 06:00 AM

presently recorded was Anatidae (14.71) > Ardeidae

to 10:00 AM and evening 04:00 PM to 06:00 PM by

(13.24) > Charadriidae (10.29) > Alcedinidae (7.35) =

walking. Study of wetland birds around the BRP area

Motacillidae (7.35) > Rallidae (5.88) = Jacanidae (5.88)

was carried out from February 2008 to January 2010,

= Threskiornithidae (5.88) > Accipitridae (4.41) >

every month at regular interval by direct counting

Phalacrocoracidae

method (Colin et al., 1993; William, 1997). The

Scolopacidae

residential status and abundance criteria was calculated

(2.94)

using presence and absence scoring method and then

>Podicipedidae (1.47) = Recurvirostridae (1.47) =

1055

(2.94)

Ciconiidae

(2.94)

(2.94 ) = Laridae (2.94 ) = Alaudidae Corvidae

(2.94)

Ploceidae

(2.94)

Journal of Research in Biology (2013) 3(6): 1054-1059

Dayananda, 2013 Table 1. Wetland bird diversity around the Bhadra Reservoir Project Area Sl. No

Order

Family

Common Name

Scientific Name

Podicipediformes

Podicipedidae

Little Grebe

Tachybaptus ruficollis

Pelecaniformes

Phalacrocoracidae

Little Cormorant

Ciconiiformes

Ardeidae

Threskiornithidae

Ciconiidae Anseriformes

Anatidae

Accipitridae

Gruiformes

Rallidae

Phalacrocorax niger

Oriental Darter

Anhinga melanogaster

Grey Heron

Ardea cinerea

Purple Heron

Ardea purpurea

Pond Heron

Ardeola grayii

Night Heron

Nycticorax nycticorax

Cattle Egret

Bubulcus ibis

Large Egret

Casmerodius albus

Median Egret

Mesophoyex intermedia

Little Egret

Egretta garzetta

Chestnut Bittern

Ixobrychus cinnamomeus

Black-headed Ibis

Threskiornis melanocephalus

Black Ibis

Pseudibis papillosa

Eurasian Spoonbill

Platalea leucorodia

Glossy Ibis

Plegadis falcinellus

White-necked Stork

Ciconia nigra

Open-bill Stork

Anastomus oscitans

Lesser-whistling Teal

Dendrocygna javanica

Common Teal

Anas crecca

Spot-billed Duck

Anas poecilorhyncha

Garganey

Anas querquedula

Nakta or Comb Duck

Sarkidiornis melanotos

Shoveler

Anas clypeata

Cotton Teal

Nettapus coromandelianus

Mallard

Anas platyrhynchos

Pintail

Anus acuta

Brahminy Duck

Tadorna ferruginea

Common Pariah Kite

Milvus migrans

Brahminy Kite

Haliastur indus

Ring tailed fishing Eagle

Haliaeetus leucoryphus

White-breasted Water hen Indian Moorhen

Amaurornis phoenicurus

I,G

Gallinula chloropus

Purple Moorhen

Porphyrio porphyrio

Common Coot

Fulica atra

Journal of Research in Biology (2013) 3(6): 1054-1059

1056

Dayananda, 2013 Charadriiformes

Jacanidae

I/G

Amaurornis akool

Water Cock or Kora

Gallicrex cinerea

Red-wattled Lapwing Yellow-wattled Lapwing

Vanellus indicus

Vanellus malabaricus

Little-ringed Plover

Charadrius dubius

Grey Plover

Pluvialis squatarola

Curlew

Numenius arquata

Common Sandpiper

Actitis hypoleucos

Recurvirostridae

Marsh Sandpiper Black-winged Stilt

Tringa stagnatilis Himantopus himantopus

Scolopacidae

Painted Snipe

Rostratula benghalensis

Bronze-winged Jacana

Metopidius indicus Hydrophasianus chirurgus

Pheasant-tailed Jacana Brown Crake Charadriidae

Common Snipe

Gallinago gallinago

Indian River Tern

Sterna aurantia

Common Tern

Sterna hirundo

Lesser-pied Kingfisher

Ceryle rudis

Small-blue Kingfisher

Alcedo atthis

Blue-eared Kingfisher White-breasted Kingfisher

Alcedo meninting

Halcyon smyrnensis

Stork-billed Kingfisher

Pelargopsis capensis

Crested Lark Black-bellied Finchlark

Galerida cristata

Eremopterix griseus

Sturnidae

Indian Myna

Acridotheres tristis

Corvidae

House Crow

Corvus splendens

Jungle Crow

Corvus macrorhynchos

Large pied Wagtail

Motacilla maderaspatensis

White Wagtail

Motacilla alba

Yellow Wagtail

Motacilla flava

Yellow-headed Wagtail

Motacilla citreola

Paddy Field Pipit

Anthus novaeseelandiae

Baya weaver bird

Ploceus philippinus

Black breasted weaver bird

Ploceus benghalensis

Laridae Alcedinidae

Alaudidae

Motacillidae

Ploceidae

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. Sturnidae (1.47) (Fig. 1). The Anatidae and Ardeidae had

The diversity may be attributed the moderate

more than nine species, this can be considered as good

volume of water storage, availability of food sources

indicators of the health of these wetlands.

(fish, crustaceans, invertebrates, aquatic plants and plankters), shelter and assured protection to these birds.

1057

Journal of Research in Biology (2013) 3(6): 1054-1059

Dayananda, 2013

Fig. 1. Percent composition of avian families represented by species richness of waterbirds around BRP area The family Anatidae dominated the list with ten species,

around the wetlands. The present work is in conformity

which was represented 14.71% of the total number of

with the earlier work of Dayananda (2008) carried out in

wetland birds present in the study area. Ardeidae was

Ramanakere of Davanagere district. Similarly, this

represented by nine species with a relative abundance of

results were in agreement with the earlier works of

13.24%. Charadriidae was represented by seven species.

Rajashekara and Venkatesha (2011); Lameed, 2011;

Motacillidae and Alcedinidae were represented by five

Bhatnagar et al., (2008) who also reported the varying

species. Threskiornithidae, Rallidae, Jacanidae were

diversity of birds in different lakes due to different

represented by four species. Accipitridae was represented

habitat conditions for roosting, nesting, feeding and

by three species and Phalacrocoracidae, Ciconiidae,

availability of food sources.

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).

REFERENCES Ali S. 1996. The Book of Indian Birds. BNHS, Oxford University Press, Mumbai. Ali S, Ripley SD and Dick JH. 1995. A Pictorial Guide to the Birds of the Indian Subcontinent, Oxford University Press, Delhi, 165.

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

Anon. 1990. Wetlands of India - A Directory. Government of India, Forests and Environment Department, New Delhi.

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 Journal of Research in Biology (2013) 3(6): 1054-1059

Bhatnagar SP, Shukla SK and Bhatnagar M. 2008. An annotated checklist of waterfowl of the Didwana inland saline lake, Nagaur, Rajasthan, India, Newsl. For 1058

Dayananda, 2013 Bird Watchers, 48:53-55. Colin J Bibby, Niel D. Burgess and David A Hill. 1993. Birds census Techniques. Academic press Ltd. London. Dayananda GY. 2008. Status of waterbirds at Ramanakere, Cheelur, Davanagere District, Karnataka. J.Aqua.Biol., 23(2): 32-35. Dayananda GY. 2009. Avifaunal diversity of Gudavi Bird Sanctuary, Sorab, Shimoga, Karnataka. Our Nature, 7:100-109. Fleming RL(Sr), Fleming RL(Jr) and Bangdel LS. 2000. Birds of Nepal with reference to Kashmir and Sikkim. First Adarsh Impression, Gaurav Offset, Delhi. Grimmett R, Inskipp C and Inskipp T. 2001. Pocket Guide to the Birds of the Indian Subcontinent. Oxford University Press, New Delhi. Huston MA. 1994. biological diversity, Cambridge University Press, Cambridge. 3rd Edition.,78. Inskipp C and Inskipp T. 1991. A Guide to the birds of Nepal. Christopher Helm. London.

Ahmadpour and Younes Yaghobzadeh. 2011. A three years study of the diversity and density of waterfowl and waders in Sorkhrud International Wetland (October 2007 â&#x20AC;&#x201C;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.

Kazmierczak K and Perlo BV. 2000. A field Guide to the Birds of the Indian Subcontinent. Yale University Press, New Haven and London. Lameed GA. 2011. Species diversity and abundance of wild birds in Dagona-Waterfowl Sanctuary Borno state, Nigeria. Afr. J. Environ. Sci. Technol., 5(10): 855-866. Manakadan R and Pittie A. 2001. Standardized common and scientific names of the Birds of the Indian subcontinent. Buceros, 6(1) : 1-37. Mc Kinnon J and Philips K. 1993. A field guide to birds of Sumatra, Java and Bali. Oxford University Press, Oxford.

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Original Research

Journal of Research in Biology

Preliminary investigations on quantity and proximate quality of maggots produced from four different sources of livestock wastes Authors: ABSTRACT: Afamdi Anene1, Olivia C. Afam-Anene2, Kelechi Ike1 Maggot, housefly larva was grown on four substrates namely: poultry (layer) and Nnamdi H. Ekekwe1 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 Institution: and 2.97g for dry weight respectively. Proximate compositions of the maggots were 1. Animal Nutrition determined using standard methods. Results indicate that the crude protein content Laboratory, Department of of the maggots ranged from 55.4% in maggots grown on pig dung to 57.42% in Animal Science/Fisheries, maggots grown on cattle blood. The crude fibre contents ranged between 0.32% and Abia State University, 0.21%. Maggots produced from pig dung and cattle blood recorded the highest ash Umuahia Campus. Abia content and the values were 11.09% and 11.20% respectively. Moisture content was State, Nigeria. 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%. 2. Department of Nutrition Significant differences (p<0.05) in the proximate composition of the maggots were and Dietetics Imo State only observed in the crude fiber, ash and moisture contents. The results from this University, P. M. B. 2000, Owerri, Imo State, Nigeria. study showed that the substrates used can produce substantial quantities of maggots with varying degrees of success. Corresponding author: Afamdi Anene.

Keywords: maggots, proximate quality, livestock wastes

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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

Web Address:

Dates: Received: 07 Aug 2013

http://jresearchbiology.com/ documents/RA0374.pdf.

Accepted: 12 Aug 2013

Published: 18 Oct 2013

Journal of Research in Biology An International Scientific Research Journal

1060-1065 | JRB | 2013 | Vol 3 | No 6

www.jresearchbiology.com

Anene et al., 2013 Pig manure, wheat bran, cattle gut and rumen

INTRODUCTION Feed is known to be the single most expensive

contents, fish guts and cattle blood are some of the

factor in animal and aquaculture production of which

substrates that have been reportedly used for the

protein is the feed constituent with the highest cost

production of maggots (Viroje et al., 1988; Ekoue et al.,

implications (Aniebo et al., 2008). Plant protein sources

2000; Aniebo et al., 2008; Ossey et al., 2012; Zhu et al.,

as alternative non-conventional protein have their

2012). However, there is a lacuna of information on the

limitations largely due to

the presence of secondary

comparative advantage in quantity of production of these

metabolites such as alkaloids, glycosides, oxalic acids,

substrates. There is a dearth of information on the

phytates, protease inhibitors, haematoglutinin, saponins,

production potentials of different substrates for the

cyanoglycosides and linamarin etc to mention a few.

production of maggots.

Plant protein sources have the advantage of low cost

This study is aimed at a comparative evaluation of;

implications as well as rich nutrient levels (Sogbesan,

The quantity of maggots harvested from poultry

2006, Sogbesan et al., 2006). These anti-nutritional

droppings, pig dung, cattle dung and cattle blood,

factors negate growth and other physiological activities

without any additional fly attractants and without

at higher inclusion levels (Oresegun and Alegbeleye,

absorbents,

2001). Fish meal which is the guaranteed protein feed

The proximate quality of the maggot so produced

ingredient in animal diets and it costs as much as

from these livestock wastes (substrates).

$2.1 per kilogram, approximately N300/kg which is about thrice the cost of soya bean meal and four times

MATERIALS AND METHODS

the cost of groundnut cake (GNC) (Aniebo et al., 2008).

The experiment was carried out at the Teaching

Consequently there is a drive to develop other protein

and Research Farm, Abia State University, Umuahia

sources too. Maggot meal has been reported to possess

Location. The treatments consisted of 30 kg each of

good nutritional value, cheaper and less tedious to

poultry droppings, cattle and pig dung; and congealed

produce than most other sources of animal protein.

blood. These were replicated three times giving each

Housefly maggots have been used as protein ingredients

replicate a weight of 10 kg and randomly placed in a

in fish feeds (Aniebo et al., 2008), poultry feeds (Inaoka

roofed open space. The exposed substrates attracted

et al., 1999, Adeniji, 2007, Hwangbo et al., 2009) and

houseflies which laid eggs that hatched into larva called

crustaceans (Cao et al., 2012).

maggots. Each substrate was sprinkled with half a liter of

The housefly (Musca domestica Linnaeus 1758)

untreated borehole water for a period of four days to

is the most common fly species and belongs to the

prevent desiccation.

phylum Insect and order Diptera. The larval forms

Harvesting

(maggots) of houseflies feed on decaying organic matter

Harvesting was done on the 4th day using the

thereby giving them the ability to degrade wastes into

sedimentation technique. Each replicate was mixed with

valuable biomass that are nutrient (fat, protein etc) rich.

7-10 liters of water and allowed to stand for 10 minutes

Many studies (Akpodiete et al., 1993, Awoniyi and

to completely separate the maggots from the substrates.

Aletor, 2002, Teguia 2005, Aniebo et al., 2008) have

Upon mixing, the substrates sank while the maggots

been conducted on the production of housefly biomass in

floated and were collected using a 3mm sieve. Harvested

simulated environments with a view in utilizing such as

maggots were taken to the laboratory for weight

feed for farm animals.

measurements and chemical analyses.

1061

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

Anene et al., 2013 Data Collection, Sample and Data Analysis Maggots from each replicate were weighed to the

Table 1: Weight of maggots produced from different livestock wastes

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

Mean Yield (g) per Kg (N=3)

Treatments Poultry Droppings Pig dung Cattle dung Cattle Blood

Wet Weight 58.730±0.34a 08.180±0.22d 12.920±0.16c 21.770±0.31b

Dry Weight 12.79±0.22a 02.97±0.17d 04.18±0.52c 07.79±0.41b

Means in the same column with different superscripts are significantly different (p<0.05).

fiber (CF), ash, nitrogen free extract (NFE), and moisture using methods described by AOAC (1995). All data were

shown that ammonia, is an effective oviposition

subjected to Analysis of Variance (ANOVA) using SPSS

attractant.

version 17 and differences in means were separated using Duncan’s Multiple Range Test (Duncan, 1955).

The results obtained in this study compared favorably with some literature reports on maggot production (Akpodiete et al., 1993, Awoniyi and Aletor,

RESULTS AND DISCUSSIONS

2002). It is important to note that the quantities of

The wet and dry weights of maggots produced

maggot produced in this study were generally lower than

from the four different wastes are presented in Table 1.

those reported in Aniebo et al., (2008). Aniebo et al.,

The result from this study shows that 1kg each of poultry

(2008) used absorbent material namely wheat brain, rice

manure, pig dung, cattle dung and congealed blood

dust and saw dust and these may have accounted for by

yielded a mean wet weight of 58.73, 8.18, 12.92 and

the higher harvests of

21.77 g of maggot. Similarly, the dry weight of maggot

(Akpodiete et al., 1993, Awoniyi and Aletor, 2002,

yield from the 1kg of the four substrates were 12.7 g

Aniebo et al., 2008) agree that the quantity of maggot

from poultry droppings, 2.97 g from pig dung, 4.18 from

produced was primarily dependent on the nature of the

cattle dung and 7.79 g from congealed cattle blood.

substrate.

maggots. These report however

These results showed that there were significant

Other factors such as moisture control and

differences (p>0.05) in the weights of maggots (wet and

inadequate aeration of substrates may influence the

dry) produced from the wastes. The trend in the quantity

quantity of maggot yield from the substrates (Calvert

of maggot production was as follows: Poultry droppings

et al., 1971). Aniebo et al., 2008 reported that high

> Cattle blood > Cattle dung > Pig dung. Insects have

density of substrates decreased aerobic conditions which

been shown to exhibit marked preferences for particular

could adversely affect the development and survival of

substrates for oviposition (Zvereva and Zhemchuzhina,

both of eggs and hatched larvae.

1988). Similarly, sites for oviposition can be influenced

Table 2 summarizes the proximate composition

by many factors among which are moisture, nutritive

of maggots produced from the different livestock wastes.

value of the substrate and the presence or absence of an

Crude protein content ranged between 55.54% in

oviposition attractant. In this study poultry manure

maggots produced from pig dung to 56.25% in maggots

characterized by high ammonium levels produced the

produced from poultry droppings and did not indicate

highest quantity of manure. (Pastor et al., 2011) have

any significant difference (p>0.05) amongst the various

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

1062

Anene et al., 2013 Table 2: Proximate composition of maggots produced from different livestock wastes Parameters Crude Protein Crude fibre Ash Ether Extract Nitrogen Free Extract

Poultry droppings 56.25±0.21a 00.32±0.08a 10.80±0.17b 22.32±0.09a 00.17±0.04a 10.12±0.11b

Moisture content

Treatments Pig dung Cattle Dung 55.54± 0.15a 56.00a± 0.00 00.26±0.05ab 00.21± 0.01b 11.09±0.15a 10.90± 0.12b a 22.64±0.07 22.66± 0.21a 00.61±0.07a 00.07± 0.01a

Cattle Blood 57.42a±0.00 00.29±0.06ab 11.20±0.11a 21.06±0.19a 00.16±0.07a

09.84±0.12b

09.86±0.16b

10.14± 0.21a

* abc: Means along the same row with different superscripts are significant (p<0.05). substrates. The crude protein content of housefly

There were significant differences (p<0.05) in

maggots has been shown by various workers to vary

ash content of maggots reared on various substrates. Ash

between 40 and 60% (Inaoka et al., 1999, Heuzé and

content of maggots reared on pig dung was 11.09% and

Tran; 2013). Hwangbo et al., (2009) recorded a protein

those reared on cattle blood was 11.20%. These values

content of 63.99% in maggots grown on chicken

were significantly lower (p<0.05) than the ash content of

droppings sprinkled with powdered milk and sugar.

maggots reared on poultry manure (10.8%) and pig dung

Lower protein regimes of 45% - 48% were reported by

(11.09%). These results on ash content of maggots differ

Fasakin et al., (2003). It is possible that higher protein

from a value of 2.74% reported for larvae of dung beetle

values in maggots may be attributed to the higher

(Aphodius rufipes) (Paiko et al., 2012) but are in tandem

nutritional content of the substrate.

with those published by Hwangbo et al., (2009). Ash

Table 2 also shows the ether extract content of maggots produced

from

various

substrates.

This

content is an indication of the mineral content of feed materials.

parameter ranges from 27.06-22.66% and did not vary

The crude fiber content of the maggots from all

significantly (p>0.05) with the substrate type. Inaoka

the substrates were all less than 1%. Similarly, there

et al., (1999) recorded crude fat content of 20% in

were significant differences in the crude fiber content.

maggots while some other authors have reported a highly

These low values indicate that maggot meal is not a good

variable lipid contents ranging between 9-25% (Heuzé

source of fiber. Similar low values ranging between

and Tran; 2013). The results of this study on the fat

0.16% for cattle blood and 0.61% for pig dung were

content of maggot produced from different substrates

recorded for nitrogen free extracts (NFE). There were no

were in tandem with those of other authors. Drying

significant differences (p<0.05) in the values obtained

methods (sun drying and oven drying) have been shown

for this parameter.

to influence the ratio of protein to fat ratio (Aniebo and Owen, 2010). Heuzé and Tran (2013) observed that fatty

CONCLUSION AND RECOMMENDATION

acid profiles of maggots are largely influenced by the

In this study, maggots of housefly larvae were

substrates on which they are grown and this may account

grown on four substrates namely: poultry (layer)

for the high variability in fat content reported by various

droppings, cattle dung, pig dung, and whole cattle blood

authors (Inaoka et al., 1999, Hwangbo et al., 2009,

in a roofed open space. The findings from this

Aniebo and Owen, 2010).

experiment showed that poultry droppings produced maggots with the highest wet and dry weights and this

1063

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

Anene et al., 2013 result may be due to the presence of ammonia in poultry

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

dropping. This study further strengthens the observation

Method of Drying on the Proximate Composition of

that the quantity of maggot produced by a substrate is

Housefly Larvae (Musca domestica Linnaeus) Meal

primarily dependent on the nature of the substrate.

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

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

2010. AOAC. 1995. Official Methods of analysis AOAC, International, P. Cunniff Sixteenth edition, Vol. II Chapter 49, 1-49. Arlington, Virginia, United States.

dung, and whole cattle blood were comparable to

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

literature reports on maggots grown on other substrates.

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

The high protein content in the maggots would greatly

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

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

Calvert CC, Martins RD and Eby HJ. 1971. Housefly pupae as food for poultry. Journal of Economic Entomology. 62(1): 939.

also further strengthens the role of maggots in

Cao JunMing, Yan Jing, Huang YanHua,

biodegradation of livestock/animal wastes and its

GuoXia, Zhang RongBin, Chen XiaoYing, Wen

importance in the management of wastes in the industry.

YuanHong,

In all, this work has provided vital information on the

replacement of fish meal with housefly maggot meal on

chemical composition of maggot meal which would

growth

facilitate its incorporation into animal and fish feeds.

immune indexes of juvenile Litopenaeus vannamei.

Zhou

performance,

TingTing.

2012.

Wang

Effects

antioxidant and non-specific

J. Fish. China, 36 (4): 529-537. REFERENCES Adeniji AA. 2007. Effects of replacing groundnut cake with maggot meal in the diet of broilers. Int. J. Poult. Sci., 6 (11): 822-825.

Duncan DB. 1955. Multiple range and multiple F tests. Biometrics, 11: 1-42. Ekoue SE and Hadzi YA. 2000. Maggots production as

Akpodiete OJ, Ologhobo AD and Oluyemi JO. 1993. The production and nutritive value of maggot meal from

proteins source for young poultry in Togo - Preliminary observations. Tropicultura, 18 (4): 212â&#x20AC;&#x201C;214.

larvae and pupal in three substrates of poultry droppings.

Fasakin EA, Balogun AM and Ajayi OO. 2003.

Report to the Dept of Animal Science, University of

Evaluation of full-fat and defatted maggot meals in the

Ibadan, Nigeria.

feeding of clariid catfish Clarias gariepinus fingerlings.

Aniebo AO, Erondu ES and Owen OJ. 2008.

Aquaculture Research. 34(9): 733-738.

Proximate composition of housefly larva (Musca

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

domestica) meal generated from mixture of cattle blood

Feedipedia.org. A programme by INRA, CIRAD, AFZ

and

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

wheat

bran.

Livestock

Research

for

Rural

Development. 20 (12):1-5.

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

1064

Anene et al., 2013 Hwangbo J, Hong EC, Jang A, Kang HK, Oh JS,

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

Kim BW and Park BS. 2009. Utilization of house fly–

2006. Harvesting techniques and evaluation of maggot

maggots a feed supplement in the production of broiler

meal as animal dietary protein source for ‘Heteoclarias’

chickens. Journal of Environmental Biology. 30(4): 609 -

in outdoor concrete tanks. World J. Agric. Sci., 2 (4):

614.

394-402.

Inaoka T, Okubo G, Yokota M, Takemasa M. 1999.

Teguia A and Beynen AC. 2005. Alternative feedstuffs

Nutritive Value of House Fly Larvae and Pupae Fed on

for broilers in Cameroon. Livestock Research for

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

Rural Development 17 (3). http://www.lrrd.org/lrrd17/3/

36 (3): 174-180.

tegu17034.htm.

Oresegun A and Alegbeleye WO. 2002. Serum and

Viroje W and Malin S. 1989. Effects of fly larval meal

Tissue

grown on pig manure as a source of protein in early

Thiocyanate

concentration

Tilapia

(Oreochromis niloticus) Fed Cassava Peel Based Diets supplemented with DL – Methionine. Journal of Aquaculture in the Tropics.17(2): 93-100.

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

Ossey YB, Koumi AR, Koffi KM, Atse BC, Kouame

maggots as feed supplement and organic fertilizer by the

LP. 2012. Use of soybean, bovine brain and maggot as

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

sources of dietary protein in larval Heterobranchus

116: 485–491.

longifilis (Valenciennes, 1840). J. Anim. Plant Sci., 15 (1): 2099-2108. Paiko YB, Dauda BEN, Salau RB and Jacob JO.

Zvereva EL and Zhemchuzhina AA. 1988. On some factors influencing the Musca domestica L. fecundity. MedskayaParazitol. 58: 27–30 [in Russian].

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

Submit your articles online at www.jresearchbiology.com

domestica (Diptera: Muscidae). Eur. J. Entomol., 108

Advantages

(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.

Easy online submission Complete Peer review Affordable Charges Quick processing Extensive indexing You retain your copyright submit@jresearchbiology.com www.jresearchbiology.com/Submit.php.

1065

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

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Recent biophysical characteristics of domestic water sources in Owerri Metropolis, Nigeria. Authors: Nwachukwu MI1*, Eziuzor SC2, Duru MKC3, Nwachukwu IO1, Ukaga CN4, Udujih OS1 and Udujih GO5.

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 Institution: study of the water sources revealed borehole water to have the least colony forming 1. Department of Microbiology, units per milliliter of total heterotrophic bacterial count (THBC), total coliform count Imo State University, P.M.B. (TCC), total Salmonella-Shigella count (TSSC), and total fungal count (TFC), as against 2000, Owerri, Nigeria. the Otamiri River with the highest values. Physicochemical characteristics of water 2. Department of Microbiology, sources studied were within permissible limit of World Health Organization (WHO) Rhema University, P.M.B. standards for domestic use. The high percentage occurrence of Salmonella species 7021, Aba, Nigeria. among other bacterial genera in the studied water sources raises a health concern. 3. Department of Biochemistry, These could be behind the high incidence of diarrhoea and typhoid infections, Abia State University, P.M.B. routinely reported in the clinics within the metropolis. With these findings, there is 2000, Uturu, Nigeria. need for public water supply authority within Owerri metropolis to improve in quality 4. Department of Animal and of water distributed. The present study has shown the recent biophysical Environmental Biological Sciences, Imo State University, characteristics of domestic water sources in Owerri metropolis, Nigeria. P.M.B. 2000, Owerri, Nigeria. 5. Department of Public Health, Federal University of Technology, P.M.B. 1526, Owerri, Imo State, Nigeria.

Corresponding author: Nwachukwu MI.

Keywords: Bio-load, biophysical characteristics, infections, water sources, Owerri metropolis.

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 Email:

Dates: Received: 16 Oct 2012

Web Address: http://jresearchbiology.com/ documents/RA0297.pdf.

Journal of Research in Biology An International Scientific Research Journal

Accepted: 05 Aug 2013

Published: 11 Nov 2013

1066-1071 | JRB | 2013 | Vol 3 | No 6

www.jresearchbiology.com

Nwachukwu et al., 2013 microbiological and physiochemical characteristics of

INTRODUCTION Water of good quality is very important to health

domestic water sources in Owerri metropolis Nigeria.

and manâ&#x20AC;&#x2122;s continued existence. The potable water

This will reveal the water source or sources that could be

provision to rural and urban population prevents health

certified suitable for domestic usages.

hazards (Lemo, 2002). Hence the principal objectives of municipal water are the production and distribution of

MATERIALS AND METHODS

safe water that is fit for human consumption (USEPA,

Water collection

2001). Therefore before describing water as potable, it

Water samples from different sources which

has to be confirmed with certain physical, chemical and

include borehole, Otamiri and Nworie rivers, tap water

microbiological standards which ensure that the water is

and rainwater were collected within Owerri metropolis

potable and safe for drinking purposes (Tebutt, 1983).

and analyzed. The samples were randomly collected

However, potable water have to be free from disease

from highly dependable points where residents usually

producing microorganisms and chemical substances

would collect their water for domestic use. Samples were

deleterious to health (Ihekoronye and Ngoddy, 1985).

collected aseptically using sterilized 500 ml glass bottles

Water can be obtained from a number of sources

following

the

guideline

APHA

(1998)

and

such as streams, lakes, rivers, ponds, rain, springs and

WHO (1984) for sampling various water sources.

wells (Chukwura, 2001). Raymond 1992 says, â&#x20AC;&#x153;Clean,

However, the river water sample was collected using the

pure and safe water only exist briefly in nature and is

method of Onyeagba et al., (2004). The collected

immediately polluted by prevailing environmental

samples were labeled appropriately and transported to

factors and human activities. Water from most sources is

the laboratory in an ice cool pack for analysis within

therefore unfit for immediate consumption without

24 hours.

treatmentâ&#x20AC;?. The consequences of water borne bacterial

Bio-load study

and viral infections have been well established along

The standard methods for the isolation and

with chemical contamination, which is known to cause

identification of microorganisms as described by

some deadly effect (Edema et al., 2001; Fapetu, 2000).

Cappucino et al., (1992) and Onyeagba et al., (2004)

It is essential that water for domestic use be

were adopted in the analyses. All the samples were

examined

frequently

contamination

may

ten-fold serially diluted before being plated out using the

intermittent. And considering the global data, morbidity

spread

of diarrhoea disease is greater than 1.5 million and

heterotrophic bacteria, count (THBC) using nutrient

mortality is 4 million with more than 2 billion people

agar, total coliform count (TCC) using MacConkey agar,

being at risk. The WHO (2003) and UNICEF (2004)

total Vibrio count (TVC) using thiosulphate citrate bile

have reported that 80% of sickness and death among

salt agar, total Salmonella-Shigella count (TSSC) using

children in the world are caused by unsafe drinking

Salmonella-Shigella agar, and total fungal count (TFC)

water. Although municipal water is distributed to large

using Sabouraud dextrose agar. All the plates were

population through closed network, but very often,

incubated for 18 to 24 hours at 37oC except for

consumers are exposed to risk of water borne diseases

fungal count that was incubated for 3 to 5 days at

due to inadequate treatment of water (Antonine and

room temperature (about 26 to 32oC). Representative

Dante, 2008; Fapetu, 2000). This study therefore is

colonies

aimed

through biochemical, microscopic and macroscopic

1067

providing

recent

information

the

plate

were

technique

streaked,

triplicates

purified,

and

for

total

identified

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

Nwachukwu et al., 2013 observations

according

Gehardt

(1994)

and

most microbiological polluted among the water sources

identification based on Holt et al., (1994).

analyzed. Nworie River was the highest in total coliform

Determination of physiochemical characteristics

while tap water produced the highest value of total

Physical and chemical indices of the water sources include colour, taste, odour, pH. Iron, total

fungal count. Rain water was next to borehole water in terms of bio-load.

alkalinity, chloride, biological oxygen demand (BOD),

Statistical analysis revealed that there was

chemical oxygen demand (COD), nitrate, conductivity,

significant difference at â&#x2030;¤0.05 in the load of different

total dissolved solids (TDS) and turbidity were

microbial groups from the different water sources

determined according to standard methods described by

analyzed.

APHA (1998).

The overall percentage occurrence of the different genera of bacteria and fungi isolated from the water sources are presented in figures 2 and 3,

RESULTS Result of the bio-load of water sources analyzed

respectively.

The

bacterial

percentage

occurrence

is shown in figure 1. The result revealed that the total

revealed Salmonella (21.7%) to be highest in occurrence

heterotrophic bacteria count (THBC) ranged between

as compared to the ties of Micrococcus (4.35%),

1.5x10 to 1.5x10 cfu/ml. The total coliform count 2

Klebsiella (4.35%) and Enterobacter (4.35%) as isolated

(TCC) was in the range 1.0 to 2.0x10 cfu/ml, the total

and analyzed. The percentage occurrence of fungi genera

Samonella/ Shigella count (TSSC) ranged from 1.5 to

isolated revealed that Aspergillus (42.85%) as the highest

2.5x10 cfu/ml, the total Vibrio count (TVC) ranged 2

from 2.5 to 7.2x10 cfu/ml, and total fungal count (TFC)

and

the

ties

Cryptococcus

(14.28%)

and

Saccharomyces (14.28%) as lowest.

ranged from 2.5 to 4.0x10 cfu/ml. The findings as shown

Statistical

analysis

revealed

significant

in figure 1, make borehole water the best among the

difference at â&#x2030;¤0.05 in the percentage occurrence of

studied water sources with no Vibrio and fungal growth;

bacterial and fungal isolates analyzed from the water

and lowest in terms of bio-load. Otamiri River had the

sources.

Cell Density (cfu/mi)

highest bio-load in the present study. This makes it the

Water Samples *A-borehole, B-Otamiri river, C-Nworie river, D-tap water, E-rainwater Figure 1. Bio-load of different water sources analyzed recently in Owerri metropolis, Nigeria. Journal of Research in Biology (2013) 3(6): 1066-1071

1068

Nwachukwu et al., 2013

Percentage occurence (%)

25 20 15 10 5 0

Bacteria genera Bacteria genera Figure 2. Overall percentage occurrence of different bacterial genera isolated from water sources in Owerri metropolis, Nigeria. The physicochemical characteristics analyzed are

objectionable, while that of Nworie was objectionable.

shown in table 1. The water sources had pH near

The overall result showed that values for most

neutrality in the range of 6.70 to 6.92. The borehole,

physicochemical indices considered in this study were

Otamiri, tap water and rainwater water sources were all

within the permissible limit as stipulated by WHO.

colourless. The colour and taste of borehole, Otamiri, tap water

and

rainwater

water

sources

were

not

Table 1. Physicochemical characteristics of water sources in Owerri metropolis Parameters Colour (TCU) ( Units) Odour Taste pH Conductivity (µs/cm) Turbidity ( NTU) Alkalinity (mg/ l) Chlorine (mg/l) Total Iron (mg/ l) BOD (mg/l) COD (mg/l) TDS (mg/l)

Water sources A B colour less colour less no no no ob 6.7 6.92 146.2 23.6 1.0 20.37 0.0 2.00 0.0 0.00 ≤0.1 ≤0.1 1.3 1.38 121.45 137.18 0.2 11.7

C dull ob ob 6.86 45.5 7.77 5.00 0.00 ≤0.1 1.48 137.18 11.7

D colour less ob ob 6.92 28.4 00.0 04.0 00.0 ≤0.1 01.2 120.2 0.1

E colour less no No 6.82 3.4 1.5 1.0 0.0 ≤0.1 1.3 117.58 0.1

Tolerance WHO 500 no no 7.0 - 8.50 500 50 600 200 0.1 2.0 196 -

TCU-true colour unit, no-not objectionable, ob-objectionable, NTU-nephlometric turbidity units. A-borehole, B-Otamiri, C-Nworie, D-tap water, E-rainwater 1069

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

Nwachukwu et al., 2013 45 Percentage occurence (%)

35 30 25 20 15

10 5

0 Cryptococcus sp.

Candida sp.

Saccharomyces sp.

Aspergillus sp.

Fungal genera

Figure 3. Overall percentage occurrence of different fungal genera isolated from water sources in Owerri metropolis, Nigeria. DISCUSSION

(WHO, 2003). From the observed results, only borehole

The water sources in Owerri metropolis as

water source was acceptable while Otamiri River,

analyzed have shown a best option in recent times for

Nworie Rivers, tap water and rain water sources were

domestic usage. The borehole water source has the least

unacceptable for domestic and drinking purposes. This

bio-load and chemical components thereby making it the

study affirms a previous study, which revealed that

best source of water for domestic use among the water

borehole water source has a good water acceptable

sources studied. This observation could be behind the

quality, microbiologically (Nwachukwu and Otokunefor,

high rate of sinking of borehole wells within Owerri

2006).

metropolis in recent times. Its low bio-load could be

The high percentage occurrence of Salmonella

attributed to the fact that it is a ground water and there is

species among other bacterial genera is a strong causal

low infiltration of pollutants from the top soil

agent. The observed high percentage occurrence of

downwards through capillary action (Chukwura, 2001;

Salmonella species in the studied water sources could be

Edema et al., 2001). Rain water which is supposed to be

associated to high diarrhoea and typhoid infections that

the cleanest source of water by nature was the second

are routinely reported in the clinics within Owerri

best in the present study. The observed low bio-load of

metropolis.

rain water could be due to the purification process that takes place during condensation while its relegation to

CONCLUSION

second best could be due to incessant and reckless air

Physicochemical characteristics of the water

pollution from diverse sources (Nwachukwu and

sources in this study fall within WHO standards for

otukunefor, 2006; Fapetu, 2000).

domestic use whereas the observed bio-load of the water

The WHO standard for domestic water supplies

sources followed the order Otamiri River > Nworie River

which recommends a 100 cfu/ml or less for total

> tap water > rain water > borehole. Borehole was the

heterotrophic bacterial count and a zero coliform per

best among the studied water sources. As inhabitants of

100ml of water was compared to results of this study

Owerri metropolis glamour for improvement in public

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

1070

Nwachukwu et al., 2013 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.

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.

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

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

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

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

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.

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.

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.

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

Journal of Research in Biology

An International Scientific Research Journal

Original Research

Journal of Research in Biology

Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae and the moth, Philosamia ricini Authors: Bendang Ao* and Sentimenla.

ABSTRACT:

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

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 â&#x20AC;&#x201C; 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.

Corresponding author: Bendang Ao.

Keywords: Mucopolysaccharides, Rhabdome.

Email:

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

Web Address:

Dates: Received: 13 Mar 2013

http://jresearchbiology.com/ documents/RA0340.pdf.

Accepted: 21 Sep 2013

Published: 11 Nov 2013

1072-1085 | JRB | 2013 | Vol 3 | No 6

www.jresearchbiology.com

Bendang, 2013 units are usually N-acetylated and sometimes N-sulfated,

INTRODUCTION Kennedy and White (1983) introduced the term “mucopolysaccharides”

describe

2-amino-2-

while the acidic units are sometimes O-sulfated (Kennedy and White, 1983).

deoxyhexose containing polysaccharides of animal origin

It is to be noted that glycosaminoglycans always

and occurring either as free polysaccharides or as their

come

protein derivative. They can be those that contain uronic

irrespective of the ways in which the term has been used,

acid

Acid

and it is now known that glycosaminoglycans are

mucopolysaccharides (AMPs) come under the second

attached covalently to proteins. Therefore, AMPs

class. Acid mucopolysaccharides (AMPs) may be further

actually refer to glycosaminoglycans of a proteoglycan

sulphated (SMP) or non sulphated e.g., chondroitin

plus, sometimes a few amino acid units.

and

those

that

are

neutral.

sulphate and hyaluronic acid respectively. These terms i . e. ,

AMPs

an d

mucopolysaccharides

category

Presence of acid mucopolysaccharides in the visual system of vertebrates are well documented. For

mucopolysaccharides) appear to provide an adequate

example, they have been reported in the bovine cornea

description and also have the added advantage of

(Coster et al., 1987; Funderburgh et al., 1996; Corpuz

continuous use (Jaques, 1977).

et al., 1996; Plaas et al., 2001; Achur et al., 2004 and

(1938)

(sul pha t ed

the

a ci d

Meyer

SMPs

within

coin ed

the

term

Conrad et al., 2010), in the eye of rabbit (Yue et al.,

“mucopolysaccharides” to include all substances with

1984; Lutjen Drecoll, 1990;

similar

physico-chemical

connective

tissues.

properties Later

“gl yc osa m in ogl yca n s”

on,

Fitzsimmons et al., 1992;

isolated

from

Takahashi et al., 1993; Goes et al., 1999; Kato et al.,

the

terms

1999), in chick cornea (Conrad et al., 1977; Li et al.,

“gl yc opr ot ei n s”

and

1992; Mc Adams and McLoon 1995),

in human and

“mucoproteins” were used, but they failed to distinguish

rabbit cornea (Freund et al., 1995; Tai et al., 1997), in

between

antibiotics

calf lens capsule (Mohan and Spiro 1991), and in the

containing amino sugars. But these terms are still found

corneal stroma of squid (Anseth, 1961 and Moozar and

in literature.

Moozar, 1973).

bacterial

polysaccharides

and

Compound eyes of insects include the lens

Other visual apparatuses where AMPs have been

system, a retina and underlying optic ganglia. Lens is a

reported are in the cornea of elasmobranchs (Balazs,

modified cuticle and is composed of the cornea and

1965), vitreous body of the eye of squids (Balazs et al.,

underlying crystalline cone. Immediately behind the

1965), in aqueous and ciliary body (Cole, 1970;

crystalline cone are the longitudinal sensory elements or

Schachtschabel

the retinula cells. The inner sides of the retinula cells

surrounding the photoreceptor cell of the cattle (Berman

collectively secrete an internal light trapping rod-like

and Bach, 1968; Berman, 1969), inter photoreceptor

structure known as rhabdom.

matrix of vertebrate (Rolich, 1970), sclera of ox (Robert

Carn ey

(1994)

1977),

interstitial

matrix

and Robert, 1967) etc. In the case of insects, AMPs have

biological

also been reported in the compounds eyes of Periplaneta

functions conferred upon them because of specific

americana, Belostoma sp (Dey, 1976), Palaemon sp,

sequences

Limulus polyphemus and Macrobrachium birmanicum

within

may have the

indi ca t ed

al.,

that

glycosaminoglycans

ha d

specific

carbohydrate

chain.

“Glycosaminoglycan” is the systematic name for the

(Dey et al., 1978),

carbohydrate residues which form linear chains of

indica (Dey, 1980)

Musca domestica, Apis cerana

alternating acidic and basic monosaccharides. The basic 1073

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

Bendang, 2013 Acid

mucopolysaccharides

play

several

hours at room temperature. Then the pH was brought to 6

important physiological roles owing to their capacity to

by the addition of HCl and the mixture was centrifuged

bind and hold water (Ogston, 1970; Ogston and Wells,

for 15 minutes at 3000rpm. To the supernatant, 0.1 ml of

1972; Wells, 1973b). They serve as natural lubricants in

2M NaCl and two volumes of ethanol were added and

the joints, impart elasticity to connective tissue, and are a

kept overnight at 5°C. The mixture was centrifuged for

component of cartilage and ligaments. They are also

15 minutes at 3000 rpm and the precipitate was collected

involved in support and motor functions, and also have

and dried. The resultant powder was re-suspended in 1

bactericidal properties. It is also known that many

ml of 0.05M sodium acetate (pH 6.5) along with 1 mg of

diseases such as collagenosis, mucoplysaccharidosis, and

DNAase and RNAase. The solution was again incubated

rheumatism etc which are correlated with aging, are also

for 24 hours at 37°C with a layer of toluene. After

a result of disorders in mucopolysaccharides metabolism

incubation, 0.1 ml of 2M Nacl and two volumes of

which lead to compositional changes of connective tissue

ethanol were added to the solution and kept overnight at

and of the body fluids.

5°C. It was then centrifuged for fifteen minutes at 3000

With this view a study was done in the compound

eye

the

insects

viz.,

rpm and precipitate was collected and dried. The

butterfly,

resultant powder was dissolved in 0.5 ml of water, heated

Pieris brassicae and moth, Philosamia ricini with

at 100°C for two minutes and analyzed by paper

regards to the occurrence of acid mucopolysaccharides,

chromatography and electrophoresis.

and their possible functions in the eyes have been

Chromatography:

discussed.

The extract was hydrolyzed with 6N HCl at 100°C for 12 hours. The acid hydrolysate was then

MATERIALS AND METHODS The eyes were separated from live insects and

evaporated to dryness. The dried residue was then dissolved in 0.5 ml of distilled water and spotted in

fixed in 10% buffered formalin until they were used.

whatman No 1 filter paper and ascending paper

Histochemical study:

chromatograms were run using butanol, acetic acid and

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

water in the ratio of 4:1:1 (v/v) as solvent (Giri and Nigam, 1954). The chromatogram was developed with silvernitrate (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

for three hours and dried. The tissues were then

hydroxide for 10 minutes and then washed in running

homogenized and suspended in 20 ml of 0.05M Tris-HCl

water and dried at room temperature.

buffer (pH 8). To the mixture, 10 mg of trypsin was

Electrophoresis:

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

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

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

1074

Bendang, 2013 electrophorectic apparatus, the paper strips were dried at

Electrophorectic movement pattern of the crude

room temperature and stained with Toluidine blue

extracts of the acid mucopolysaccharides from the eyes

(0.04% in 80% acetone). The staining of the strips was

of Pieris brassicae and Philosamia ricini, when

followed by 2-3 rinsing in 0.1% acetic acid and then 2-3

com par ed

times in H2O. The strips were then dried at room

mucop ol ysa cch ar ides

temperature.

mucopolysaccharides extracted resemble chondroitin

OBSERVATIONS

4-sulfate (Figure 5 and 6; Table 3 and 4).

with

several

standard

sh owed

that

a ci d th e

Histochemical observations: Lens cuticle of the butterfly, Pieris brassicae:

DISCUSSION

When the sections of the eyes were stained with

Several workers like Miao et al., (1996), Groves

toluidine blue, the cornea and crystalline cone became

et al., (2005), Manton et al., (2007), Fthenou et al.,

purple in color showing metachromasia (Photoplate 1)

(2006, 2008) etc. have studied the influence of

i.e.,

acid

glycosaminoglycans on cell division, differentiation,

mucopolysaccharides, while the region of the rhabdom

responses to growth factors, adhesion, migration,

was

orthochromatic (blue in colour) and therefore

peripheral nerve extension or regeneration and signal

devoid of acid mucopolysaccharides. Similarly, when the

transduction. In this regard, Bulow and Hobert, (2006)

eyes were stained with alcian blue, the lens and

are of the opinion that the correct development of a

crystalline cone became purple in colour (Photoplate 2)

multicellular organism is via a specific code contributed

wh i ch

by the glycosaminoglycans.

i n di c a t i n g

i n di ca t e s

th e

pr e s e n c e

the

pr e s en c e

a ci d

mucopolysaccharides. (Fig 1)

In the case of the visual apparatus, they play a central role in the physiological maintenance of

Lens cuticle of the moth, Philosamia ricini: When the sections were stained with toluidine

trabecular meshwork in the eyes (Yue et al., 1984 and

blue, the cornea as well as crystalline cone became

Cavallotti et al., 2004). They may also have a role in

purple in colour (Photoplate 3) showing the presence of

influencing keratocytes and nerve growth in corneal

mucopolysaccharides. The more intense reactions were

stroma because of their ability to bind together (Cornard

observed towards the corneal lens. The rhabdom region

et al., 2010). They, and their core proteins also have

however gave a blue colour reaction i.e. the region is

important physiological and homeostatic roles e.g.

orthochromatic (Photoplate 4). When the eyes were

during inflammation and immune response (Park et al.,

stained with alcain blue the corneal lens and crystalline

2001; Li et al., 2002; Wang et al., 2005).

cone became purple in colour indicating the presence of

AMPs influence tissue osmotic pressure not only

AMPs, while the rhabdom became blue in colour which

by influencing the water balance, but also by introducing

indicates absence of AMPs. (Fig 2)

excess swelling pressure which is balanced by an internal

Biochemical observations:

structural resistance (Ogston, 1970). Moreover, AMPs

Chromatographic

acid

play important roles in â&#x20AC;&#x153;water bindingâ&#x20AC;? and maintenance

mucopolysaccharides extract showed the presence of

of tissue osmotic pressure (Ogston and Wells, 1972).

three sugars viz lactose, galactose and xylose in case of

Payrau et al., (1967) observed that the transparency of

Pieris brassicae and galactose, xylose and rhamnose in

the cornea is based on the state of hydration of tissue.

the case of

They based this on the fact that the corneal stroma of

Table 1 and 2). 1075

analysis

the

Philosamia ricini (Figure 3 and 4;

most vertebrates, including mammals, birds and teleosts Journal of Research in Biology (2013) 3(6): 1072-1085

Bendang 2013

Fig 1. Histochemical observations of Lens cuticle of the Fig 2. Histochemical observations of Lens cuticle of the butterfly, Pieris brassicae moth, Philosamia ricini absorb water wherever free water is accessible. In

shown that acid mucopolysaccharides possess high water

contrast, according to Maurice and Riley (1970) odema

binding capacities.

of the cornea leads to disorganization of its structure and

Multiple

types

chondroitin

sulphate

less transparency, but dehydration does not appear to

proteoglycans are seen in vertebrates and they greatly

have serious optical affects. Maurice (1972) suggested

influence development and tissue mechanics. For

that the presence of AMPs in the cornea is mainly

example, the chondroitin chains in the nematode

responsible for the dehydration properties of the tissue

Caenorhabditis elegans are not sulphated, but are

and hence transparency. This is supported by workers

nevertheless essential for embryonic development and

like Hedbys (1961, 1963); Kikkawa and Hirayama

vulval morphogenesis (Olson et al., 2006). Chondroitin

(1970); Bettelheim and Plessy (1975); Lee and Wilson

and dermatan proteoglycans have also been the subject

(1981) and Castoro et al., (1988).

of much interest as inhibitors of axon growth and have

AMPs have also been suggested to play a major

been shown to be important components of the glial scar

role in the structural organization of intracellular matrix

that prevents axon regeneration (Rhodes and Fawcett,

via electrostatic and steric interactions with other

2004).

macromolecules of the matrix, such as collagen and

The

role

mucopolysaccharides

elastin (Kobayashi and Pedrini, 1973). Similarly, Ogston

pathogenicity has been widely reviewed. For instance,

and Wells, (1972) have suggested that AMPs help in the

they are responsible for calcification of bones (Rubin and

maintenance of mechanical flexibility and elasticity of

Howard, 1950), dermal thickening in acromegalic

tissues. Ogston, (1966a) and Katchalsky, (1964) have

patients (Matsuoka et al., 1982), involved in inborn

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

1076

Bendang 2013 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 Butterfly, Pieris brassicae Moth, Philosamia ricini

Rf value 0.05 0.18 0.33 0.16 0.33 0.43

Identification Lactose Galactose Xylose Galactose Xylose Rhamnose

errors of metabolism and/ or storage disorders (Matalon et al., 1974a; Hall et al., 1978; Neufeld and Fratantoni,

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 Raffinose Lactose Glucose Sucrose Galactose Mannose Fructose Xylose Ribose

Rf value 0.03 0.05 0.10 0.13 0.18 0.25 0.28 0.34 0.38

1970; McKusick et al., 1978), maintenance of retinal

(Meenakshi and Scheer, 1959; Sundara Rajulu, 1969;

structure and neural tube closure in Knobloch syndrome

Krishnan, 1965 and Raghuvarman et al., 1998), it is

(Sertie et al., 2000) and treatment of diabetic

reasonable to presume that the specific occurrence of

nephropathy (Gambaro and Van Der Woude, 2000).

mucopolysaccharides in the lens cuticle and the

Matthews (1959) and Oosawa (1971) have

crystalline cone may have a bearing on the visual system

suggested that one of the characteristic properties of

of the insects. Keeping the above account in view it is

mucopolysaccharides is the selective association or

possible to assume a role of AMPs in the lens-cuticle of

binding with small inorganic cations, especially H , Na , and

Ca++,

and

also

with

The lens-cuticle as already stated, besides

macromolecules. In these regard, Farber and Schubert

playing a general defensive role, performs a special

(1957) and Urist et al., (1968) have also found a small

optical function of conducting light rays to the inner

over Na in chondroitin

rhabdomere. It is possible to presume that the

sulphate. Matthews (1975) thus suggested that these

transparency of the lens-cuticle, which is more than that

preference for binding Ca

substances act as a store for Ca

cationic

groups

insects.

in cartilage tissue and

of other types of cuticle (e.g. body cuticle), may be

that is the reason for their specific roles in tissue-

affected by the occurrence of mucopolysaccharides

calcification. Some roles of AMPs, especially in

(Anseth and Fransson, 1970). Similarly, Freund et al.,

arthropodan cuticle have been reported by Meenakshi

(1995) also reported that the presence of AMPs in human

and Scheer (1959) and Sundara Rajulu (1969) in terms of

and rabbit cornea is related to transparency. It is known

calcification of the cuticle of Hemigrapsus nudus and

that the bulk of cornea of vertebrate eye is the stroma,

Cingalobolus bugnioni respectively. Krishnan (1965) has

which functions as a supporting structure and is adapted

suggested that AMPs may be associated with -S-S-

for the transmission of a high percentage of incident light

bonding of the cuticle in the scorpion Palaemonetes

of visible-wave length (Maurice, 1969). Anseth and

swammerdami.

Fransson (1970) have found that during chick corneal

Since

a cid

development, the occurrence of a highly sulfated keratan

mucopolysaccharides is not a general feature of the

sulfate is associated with rise in the transparency of

arthropod cuticle and it occurs in some special types of

stroma. They have

cuticle where it performs some special functions

transparency is correlated with the presence of normal

1077

the

occurrence

also suggested that

stromal

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

Bendang 2013

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.

Fig. 5: Paper electrophorectic movement patterns of the crude mucopolysaccharides from the eyes of the butterfly, Pieris brassicae.

Fig. 6: Paper electrophorectic movement pattern of the crude mucopolysaccharides from the eyes of the moth Philosamia ricini

proportions of keratan sulfate and chondroitin 4-sulfate.

Transparency of the corneal stroma depends partially on

Funderburgh et al., (1996) have reported that

the degree of spatial order of its collagen fibrils which

keratan proteoglycans are the major proteoglycans of the

are narrow in diameter and closely packed in a regular

bovine cornea and secreted by keratocytes in the corneal

array (Maurice, 1957; Cox et al., 1970; Benedek, 1971;

stroma and they are thought to play an important role in

Mc Cally and Farrell, 1990 and Bron, 2001). Mc Adams

corneal structure and physiology, particularly in the

and Mc Loon (1995) have shown that retinal axons grow

maintenance of corneal transparency. Blochberger et al.,

in the presence of chondroitin sulphate and keratan

(1992), has reported that corneal keratan sulfate

sulfate proteoglycans and that these proteoglycans helps

proteoglycans contribute to corneal transparency in

in developing chick visual pathway.

chick. Takahashi et al., (1993) have also reported that

Many studies that focused on corneal swelling

keratan sulfate and dermatan sulfate proteoglycans are

behavior have noted a gradual decrease in swelling from

associated with collagen in foetal rabbit cornea.

the posterior to anterior side (Van Horn et al., 1975;

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

1078

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)

Table 4: Paper electrophorectic movement patterns of some standard mucopolysaccharides. (Solvent used is phosphate buffer of pH 6.5)

Bettelheim and Plessy 1975; Castoro et al., 1988 and

Standard mucopolysaccharides Heparin Chondroitin 4-sulfate Heparan sulfate Chondroitin 6-sulfate Keratan sulfate Dermatan sulfate

Cristol et al., 1992) and this was thought to be related to

(Moozar and Moozar, 1972).

Distancetravelled (cms)

Insect Butterfly, Pieris brassicae Moth, Philosamia ricini

Acid mucopolysaccharide type

6.4

Chondroitin 4-sulfate

6.8

Chondroitin 4-sulfate

Distance travelled (cms) 5.5 6.6 7.2 7.6 8.7 10.0

the organization of the collagen lamellae and the

It is well-established that one of the corneal

presence of different types of proteoglycans. In the

limiting cell layers i.e., the corneal endothelium,

posterior part, keratan sulfate, a more hydrophilic

transports fluid at a substantial rate and that this transport

proteoglycan is prevalent, whereas in the anterior part

is essential to maintain normal stromal hydration

dermatan sulfate, a much less hydrophilic proteoglycan,

(Maurice, 1972; Candia, 1976; Candia and Zamudio,

is present (Bettelheim and Plessy 1975; Castoro et al.

1995; Narula et al., 1992; Bonanno et al., 1989 and Yang

1988). An interesting conclusion was drawn by Muller et

et al., 2000). Anseth and Fransson, (1969) had

al., (2001) while studying the differential behaviour of

demonstrated the synthesis of AMPs by corneal

the anterior and posterior stroma during corneal swelling,

epithelial and stromal cells, and that they are important

that

the

in maintaining the corneal structure in relation to its

glycosaminoglycan components of the proteoglycans that

environment. Deb and Raghuvarman (1994) have also

is responsible for the corneal swelling due to electrostatic

observed that glycosaminoglycans are essential for the

repulsion between acidic groups. They also suggested

maintenance of corneal structure and function.

the

high

negative

charge

that the structural stability of the anterior stroma under

Acid mucopolysaccharides thus detected in the

condition of extreme hydration imply an important role

compound eyes of the butterfly, pieris brassicae and the

for this zone in the maintenance of corneal curvature and

moth, Philosamia ricini may play an important role in

that this stability is determined by the tight interweave of

visual excitation, when light rays pass through the outer

the stromal lamellae.

epicuticle to the inner endocuticular region (crystalline

It is now known that the pH value is a decisive

cone) - the sites of AMPs, due to the fact that they act as

factor for the taking of water by the cornea (Cejkova and

a selective ion barrier (Jeanloz, 1970). It may also be

Brettschneider,

noted that they are present not only in the corneal lens

complex

1969). The protein polysaccharide specific

but also in the crystalline cone, which are in close

configuration within the molecules than electro-static

connection with the inner rhabdomeres (the actual sites

linkage could. For the cornea to remain transparent, it is

of photochemical reactions), the products of which may

essential that an active mechanism counter the natural

depolarize the membrane of the retinula cells and initiate

tendency of the stroma to increase its hydration, swelling

impulse formation (Wigglesworth, 1965). Further,

and opacity. It may be noted here that the non - swelling

mucopolysaccharides may play a role in increasing

properties of elasmobranch cornea is supposed to be due

transparency of lens-cuticle. In this context, it is worth

to the high mannose content in their structural proteins

mentioning

1079

provides

stable

and

that

during

corneal

development

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

Bendang, 2013 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

the polysaccharides for the normal function of the corneal stroma. Acta. Ophthal. 48(3): 455 - 464. Balazs EA. 1965. Amino sugar-containing macromolecules in tissues of the eye and ear. In: The amino sugars. Balazs EA and Jeanloz RW, Eds. Vol 2A, 401 - 460. Academic press, New York.

to them.

Benedek GB. 1971. Theory of transparency of the eye. Appl. Optics. 10(3): 459 – 473.

CONCLUSIONS

Berman ER and Bach G. 1968. The acid mucopolysaccharides of cattle retina. Biochem. J. 108 (part 1): 75 - 88.

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. REFERENCES Achur RN, Muthusamy A, Madhunapantula SV, Bhavanandan VP, Seudieu Cand Gowda DC. 2004. Chondroitin sulfate proteoglycans of bovine cornea: structural characterization and assessment for the adherence of Plasmodium falciparum-infected erythrocytes. Biochem. Biophsy. Acta. 1701(1-2):109119. Anseth A. 1961. Studies on corneal polysaccharides*: 111. Topographic and comparative biochemistry. Exp. Eye. Res. 1(2): 106 - 115. Anseth A and Fransson LA. 1969. Studies on corneal polysaccharides*: V1. Isolation of dermatan sulfate from corneal scar tissue. Expl. Eye. Res. 8(3): 302 - 309. Anseth A and Fransson LA. 1970. The importance of Journal of Research in Biology (2013) 3(6): 1072-1085

Ber man ER. 1969. Mucopol ysa cch ari des (glycosaminoglycans) of the retina. Identification, distribution and possible biological role. Biol. Ophthalmol. 79: 5 - 31. Bettelheim FA and Plessy B. 1975. The hydration of proteoglycans of bovine cornea. Biochem. Biophys. Acta. 381(1): 203 - 214. Blochberger TC, Vergnes JP, Hempel J and Hassell JR. 1992. cDNA to chick lumican (Corneal keratan sulfate proteoglycan) reveals homology to the small interstitial proteoglycan gene family and expression in muscle and intestine. J. Biol. Chem. 267 (1): 347 – 352. Bonanno JA, Klyce SD and Cragoe EJ. 1989. Mechanism of chloride uptake in rabbit corneal epithelium. Am. J. Physiol. 257(2 pt 1): C290 - C296. Bron AJ. 2001. The architecture of the corneal stroma. Br. J. Ophthalmol. 85(4): 379 - 381. Bulow HE and Hobert O. 2006. The molecular diversity of glycosaminoglycans shapes animal development. Annu. Rev. Cell. Dev. Biol. 22: 375 - 407. Candia OA. 1976. Fluid and Cl transport by the epithelium of the isolated frog cornea. Fedn. Proc. 35: 703. Candia OA and Zamudio AC. 1995. Chloride-activated water permeability in the frog corneal epithelium. J. Membr. Biol. 143(3): 259 - 266. Carney SL. 1986. Proteoglycans. In Carbohydrate analysis-a practical approach. Chaplin MF and Kennedy JF, Eds. Oxford University Press, New York. 1080

Bendang, 2013 Castoro JA, Bettelheim AA, Bettelheim FA. 1988. Water gradients across bovine cornea. Invest. Ophthamol. Vis. Sci. 29(6): 963 - 968.

Deb MK and Raghuvarman A. 1994. Glycosaminoglycan fractions in the cornea of two fishes. Curr. Sci. 67(12): 1020.

Cavallotti C, Feher J, Pescosolido N and Sagnelli P. 2004. Glycosaminoglycans in human trabecular meshwork: age- related changes. Ophthalmic. Res. 36(4): 211 - 217.

Dey S. 1976. Acid mucopolysaccharides in the lens cutice of insects. MSc dissertation. North Eastern Hill University, Shillong, Meghalaya, India.

Cejkova J and Brettschneider I. 1969. The effect of hydration of the rabbit cornea on the histochemical demonstration of acid mucopolysaccharides. Histochem. 17(4): 327 - 336. Cole D. 1970. Aqueous and ciliary body. In: Biochemistry of the eye. Graymore CN, Ed. 105 - 171. Academic Press, London and New York. Conrad AH, Zhang Y, Tasheva ES and Conrad GW. 2010. Proteomic analysis of potential keratan sulfate, chondroitin sulfate A, and hyaluronic acid molecular interactions. Invest Ophthalmol. Vis. Sci. 51(9): 4500 4515. Conrad G, Hamilton C and Haynes E. 1977. Differences in glycosaminoglycans synthesized by fibroblast-like cells from chick cornea, heart and skin. J. Biol. Chem. 252(19): 6861 - 6870. Corpuz LM, Funderburgh JL, Funderburgh ML, Bottomley GS, Prakash S and Conrad GW. 1996. Molecular cloning and tissue distribution of keratocan: bovine corneal keratan sulfate proteoglycan. 37A. J. Biol. Chem. 271(16): 9759 - 9763. Coster L, Rosenberg LC, Van der Rest M and Poole AR. 1987. The dermatan sulfate proteoglycans of bovine sclera and their relationship to those articular cartilage. An immunological and biochemical study. J. Biol.Chem. 262(8): 3809 - 3812. Cox JL, Farrell RA and Hart RW and Langham ME. 1970. The transparency of the mammalian cornea. J. Physiol (Lond). 210(3): 601 - 616. Cristol SM, Edelhauser HF and Lynn MJ. 1992. A comparison of corneal stromal edema induced from the anterior or the posterior surface. Refract. Corneal. Surg. 8(3): 224 - 229.

1081

Dey S. 1980. Studies on certain physiological and biochemical aspects of vision in some arthropods. Ph.D. Thesis. North Eastern Hill University, Shillong, Meghalaya, India. Dey S, Raghuvarman A and Michael RG. 1978. Acid mucopolysaccharides in the lens cuticle of fresh water prawn, Macrobrachium birmanicum (Crustacea) and the marine horse shoe crab, Limulus polyphemus (Xiphosura) and their physiological significance. IV. All. India. Symp. On. Comp. Animal. Physiol. Endocrinol. Dietrich CP, Sampaio LO and Toledo OMS. 1977. Preparation of a crude AMPs fraction from tissues with proteolytic enzymes. In: Methods of biochemical analysis. Glick D, Ed. Vol. 24, 284 - 285. Wiley, New York. Farber SJ, and Schubert M. 1957. The binding of cations by chondroitin sulfate. J. Clin. Invest. 36 (12):1715 - 1722. Fitzsimmons TD, Fagerholm P, Harfstrand A and Schenholm M. 1992. Hyaluronic acid is in the rabbit cornea after excimer laser superficial keratectomy. Invest. Ophthalmol. Vis. Sci. 33(11): 3011 - 3016. Freund DE, McCally RL, Farrell RA, Cristol SM, L'Hernault NL and Edelhauser HF. 1995. Ultrastructuure in the anterior and posterior stroma of perfused human and rabbit corneas. Relation to transparency. Invest. Opthalmol. Vis. Sci. 36(8): 1508 1523. Fthenou E, Zafiropoulos A, Tsatsakis A, Stathopoulos A, Karamanos NK and Tzanakakis GN. 2006. Chondroitin sulfate A chains enhance platelet derived growth factor-mediated signalling in fibrosarcoma cells. Int. J. Biochem. Cell. Biol. 38(12): 2141 â&#x20AC;&#x201C; 2150. Fthenou E, Zafiropoulos A, Katonis P, Tsatsakis A, Karamanos NK and Tzanakakis GN. 2008. Chondroitin sulfate prevents platelets derived growth Journal of Research in Biology (2013) 3(6): 1072-1085

Bendang, 2013 factor-mediated phosphorylation of PDGF-Rbeta in normal human fibroblasts severely impairing mitogenic responses. J. Cell. Biochem. 103(6): 1866 - 1876. Funderburgh JL, Funderburgh ML, Mann MM, Prakash S and Conrad GW. 1996. Synthesis of corneal keratan sulfate proteoglycans by bovine Keratocytes in vitro. J. Biol. Chem. 271 (49): 31431 - 31436. Giri KV and Nigam AN. 1954. Circular paper chromatography of carbohydrates. J. Indian. Inst. Sci . 36: 49-63. Goes RM, Laicine EM, Porcionatto MA, Bonciani NH and Haddad A. 1999. Glycosaminoglycans in components of the rabbit eye: Synthesis and charcterization. Curr. Eye. Res. 19 (2): 146 â&#x20AC;&#x201C; 153.

Kato T, Nakayasu K, Ikegami K, Obara T, Kanayama T and Kanai A. 1999. Analysis of glycosaminoglycans in rabbit cornea after excimer laser keratectomy. Br. J.Ophthalmol., 83(5): 609 - 612. Kennedy JF and White CA. 1983. Bioactive carbohydrates: In Chemistry, biochemistry and biology. 183. Ellis Horwood Limited. Halstead Press, New York, Brisbane, Chichester, Toronto. Kikkawa Y and Hirayama K. 1970. Uneven swelling properties of the corneal stroma. Invest. Ophthalmol. 9 (10): 735 - 741. Kobayashi TK and Pedrini V. 1973. Proteoglycanscollagen interactions in human costal cartilage. Biochem. Biophys. Acta . 303(1):148 - 160.

Gambaro G and Van Der Woude FJ. (2000). Glycosaminoglycans: Use in treatment of diabetic nephropathy. J. Am. Soc. Nephrol. 11(2): 359 - 368.

Krishnan N. 1965. Occurance of a cid mucopolysaccharides in the cuticle of the scorpion Palamneus swammerdami. Curr. Sci. 34(4): 120 - 121.

Groves ML, McKeon R, Werner E, Nagarsheth M, Meador W and English AW. 2005. Axon regeneration in peripheral nerves is enhanced by proteoglycan degradation. Exp. Neural. 195(2): 278 - 292.

Lee D and Wilson G. 1981. Non-uniform swelling properties of the corneal stroma. Curr. Eye.Res. 1(8): 457 - 461.

Hall CW, Liebaers I, Di Natale P and Neufeld EF. 1978. Enzymic diagnosis of the genetic mucopolysaccharides storage disorders. Methods. Enzymol. 50: 439-456. Hedbys BO. 1961. The role of polysaccharides in corneal swelling. Exp. Eye. Res. 1(1): 81 â&#x20AC;&#x201C; 89. Hedbys BO. 1963. Studies on the hydration of the corneal stroma. 1 - 20. Akademisk. Lund. Humason GL. 1971. Animal tissue techniques. W. H. Freeman and Co. San Francisco. Jaques LB. 1977. Determination of heparin and related sulfated mucopolysaccharides. In: Methods of biochemical analysis. 24 - 206. 24:203-312. Jeanloz RW. 1970. Mucopolysaccharides of higher animals. In The carbohydrates. W.Pigman and Herp, D.Horton. Eds. Vol 2B. 589 - 625 Academic Press, New York. Katchalsky A. 1964. Polyelectrolytes and their biological interactions. Biophysc. J. 4(1): 9 - 41.

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

Leitner, JG and Kerby, GP 1954. Staining of acid mucopolysaccharides after chromatography on filter paper. Biotechnic & Histochemistry. Vol. 29, No. 5, 257259 Li W, Vergnes JP, Cornuet PK and Hassell JR. 1992. cDNA clone to chick corneal chondroitin dermatan sulfate proteoglycan reveals identity to decorin. Arch. Biochem. Biophys.296(1): 190 - 197. Li Q, Park PW, Wilson CL and Parks WC. 2002. Matrilysin shedding of syndecan-1 regulates chemokine mobilization and trans-epithelial efflux of neutrophils in acute lung injury. Cell. 111(5): 635 - 46. Lutjen-Drecoll E, Schenholm M, Tamm E and Tengblad A. 1990. Visualization of hyaluronic acid in the anterior segment of rabbit and monkey eyes. Exp. Eye. Res. 51 (1): 55 - 63. Manton KJ, Leong DFM, Cool SM and Nurcombe V. (2007). Disruption of heparan and chondroitin sulfate signaling enhances mesenchymal stem cell-derived osteogenic differentiation via bone morphogenetic

1082

Bendang, 2013 protein signaling pathways. Stem Cells. 25(11): 2845 2854. Matalon R, Arbogast B and Dorfman A. 1974 a. Deficiency of chondroitin sulfate ChS Nacetylgalactosamine 4-sulfate sulfatase in MaroteauxLamy syndrome. Biochem. Biophys. Res. Commun. 61: 1450-1457. Mathews MB. 1959. Macromolecular properties of isomeric chondroitin sulfates. Biochem. Biophys. Acta. 35: 9 - 17. Mathews MB. 1975. Connective TissueMacromolecular structure and evolution. SpringerVerlag, Berlin, Heidelberg, New York. 318 pages. Matsuoka LY, Wortsman J, Kupchella CE, Eng A and Dietrich JE. 1982. Histochemical characterization of the cutaneous involvement of acromegaly. Arch. Intern. Med. 142 (10): 1820 - 1823. Maurice DM and Riley MV. 1970. In: The biochemistry of the eye. Graymore CN, Ed. 1-95. Academic Press, London and New York. Maurice DM. 1957. The structure and transparency of the cornea. J. Physiol (Lond). 136(2): 263 - 286. Maurice DM. 1969. The cornea and sclera: In the eye. Davson H, 2nd Ed. Vol. 1, pp. 489-600. Academic Press, New York. Maurice DM. 1972. The location of the fluid pump in the cornea. J. Physiol. London. 221(1): 43 - 54. McAdams BD. and McLoon SC. 1995. Expression of chondroitin sulfate and keratan sulfate proteoglycans in the path of growing retinal axons in the developing chick. . 352 (4): 594 - 606. McCally RL and Farrell RA. 1990. Light scattering from cornea and corneal transparency. In: Non invasive diagnostic techniques in ophthalmology. Masters BR, Ed . New York, Springer-Verlag. pp 189 - 210. McKusick VA, Neufeld EF and Kelley TE. 1978. The mucopolysaccharides storage diseases. In : The metabolic basis of inherited disease. Stanbury JB, Wyngaarden J B. and Fredrickson, DS, 4th Eds. pp. 1282. McGraw-Hill, New York.

1083

Meenakshi VR and Scheer BT. 1959. Acid mucopolysaccharides of the crustacean cuticle. Science. 130(3383): 1189 - 1190. Meyer K. 1938. The chemistry and biology of mucopolysaccharides and glycoproteins. Cold Spring Harbor Symposia on Quantitative Biology. 6: 91-102. Miao HQ, Ishai-Michaeli R, Atzmon R, Peretz T and Vodavsky I. 1996. Sulfate moieties in the subendothelial extracellular matrix are involved in basic fibroblast growth factor sequestration, dimerization, and stimulation of cell proliferation. J. Bio. Chem. 271(9): 4879 â&#x20AC;&#x201C; 4886. Mohan PS and Spiro RG. 1991. Characterization of heparan sulfate proteoglycan from calf lens capsule and proteoglycans synthesized by cultured lens epithelial cells. Comparison with other basement membrane proteoglycans. J. Biol. Chem. 266 (13): 8567 -8575. Moczar E. and Moczar M. 1972. The phylogenetic evolution of the corneal stroma. Distribution of carbohydrates in the macromolecular complexes of the primate, Baboon (Cynocephalus papio- papio) corneal stroma. Comp. Biochem. Physiol. 43(2): 261 - 270. Moczar M. and Moczar E. 1973. Phylogenetic evolution of the cornea. Macromolecular composition of the corneal stroma of the squid (Sepia officinalis). Comp. Biochem. Physiol. 45(1): 213 - 223. Muller LJ, Pels E and Vrensen FJM 2001. The specific architecture of the anterior stroma accounts for maintenance of corneal curvature. Br. J. Ophthalmol. 85 (4): 437 â&#x20AC;&#x201C; 443. Narula P, Xu M and Kuang KY, Akiyama R and Fischbarg J. 1992. Fluid transport across cultured bovine corneal endothelial cell monolayers. Am. J. Physiol. 262 (1 pt 1): C98 - 103. Neufeld EF and Fratantoni JC. 1970. Inborn errors of mucopolysaccharides metabolism. Science. 169(3941): 141-146. New York, London. Ogston AG. 1966 a. On water binding. Federation. Proc. 25(3): 986 - 989. Ogston AG. 1970. The biological function of the glycosaminoglycans. In: Chemistry and Molecular Journal of Research in Biology (2013) 3(6): 1072-1085

Bendang, 2013 Biology of the intercellular matrix. Balazs, E. A. Ed. Vol. 3. pp. 1231 -1240. Academic Press, New York. Ogston AG and Wells JD. 1972. The osmotic properties of sulphoethylâ&#x20AC;&#x201C;sephadex. A model

for

cartilage.

Biochem. J. 128(part 3): 685 - 690. Olson SK, Bishop J R, Yates J R, Oegema K. and Esko JD. 2006. Identification of novel chondroitin proteoglycans in Caenorhabditis elegans : embryonic cell division depends on CPG-1 and CPG-2. J. Chem. Biol.

Protides of biological fluids, vol. is, p. 143. Amsterdam: Elsevier 1967. Rubin PS and Horward JR. 1950. Conference on metabolic interactions. Trans. 2nd Conf. Col. 2. 155. Schachtschabel DO, Bigalke B and Rohen JW. 1977. Production of glycosaminoglycans by cell cultures of the trabecular meshwork of the primate eye. Exp. Eye. Res. 24(1): 71 - 80. Sertie AL, Sossi V, Camargo AA, Zatz M, Brahe C

173 (6): 985 - 994.

and Passos-Bueno MR. 2000. Collagen XV111,

Oosawa F. 1971. Polyelectrolytes. Dekker, New York.

containing an endogenous inhibitor of angiogenesis and

Park PW, Pier GB, Hinkes MT and Bernfield M.

tumor growth, plays a critical role in the maintenance of

2001.

retinal structure and in neural tube closure (Knobloch

Exploitation

syndecan-1

shedding

Pseudomonas aeruginosa enhances virulence. Nature. 411(6833): 98 - 102.

syndrome). Hum. Mol. Genet. 9(13): 2051 - 2058. Sundara

Rajulu

1969.

Presence

acid

Payrau P, Pouliquen Y, Faure JP and Offret G. 1967.

mucopolysaccharides in the cuticle of a millipede,

In : Biochemisty of the eye. Graymore, C. N. Ed. pp. 13.

Cingalobolus bugnioni. Curr. Sci. 38(14): 345 - 346.

Academic press, London, New York.

Tai GH,

Nieduszynski IA, Fullwood NJ and

Plaas AH, West LA, Thonar EJA, Karcioglu ZA,

Huckerby TN. 1997. Human corneal keratan sulfates.

Smith CJ, Klintworth GK and Hascall VC. 2001. Altered

J. Biol. Chem. 272(45): 28227 - 28231.

fine structures of corneal and skeletal keratan sulfate and chondroitin/dermatan

sulfate

macular

corneal

dystrophy. J. Biol. Chem. 276(43): 39788 - 39796.

Acid mucopolysaccharides in the body cuticle of the Peripatus, Typhiloperipatus weldoni in relation to the cutaneous mode of sperm transfer. J. Endocrinol. Reprod. 1 (2): 80 - 85. Rhodes KE and Fawcett JW. 2004. Chondroitin proteoglycans:

preventing

plasticity

protecting the CNS? J. Anat. 204(1): 33 - 48. Robert I and Robert B. 1967. Structural glycoproteins of membranes and connective tissue: biochemical and immunopathological properties. In: H. PEETERS, (ed.),

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

1993. Keratan sulfate and dermatan sulfate proteoglycans associate with type V1 collagen in fetal rabbit cornea.

Raghuvarman A, Dey S and Hermann HR Jr. 1998.

sulphate

Takahashi T, Cho HI, Kublin CL and Cintron C.

J. Histochem. Cytochem. 46 41(10): 1447 - 1457. Trevelyan WE, Procter DP and Harrison JS. 1950. Detection of sugars on paper chromatograms. Nature. Lond. 166(4219): 444 - 445. Urist MR, Speer DP, Ibsen KJ and Strates BS. 1968. Calcium binding by chondroitin sulfate. Calcif. Tissue. Res. 2(1): 253 - 261. Van Horn DL, Doughman DJ, Harris JE, Miller GE, Lindstrom R and Good RA. 1975. Ultrastructure of human

organ-cultured

cornea.

11.

Stroma

and

epithelium. Arch. Ophthalmol. 93(4): 275 - 277. 1084

Bendang, 2013 Wang L, Fuster M, Sriramarao P and Esko JD. 2005. Endothelial heparan sulfate deficiency impairs L-selectin and chemokine-mediated neutrophil trafficking during inflammatory responses. Nat. Immunol. 6(9): 902 -910. Wigglesworth VB. 1965. The principle of Insect Physiology. English Language book Society. Methuen and Co. Ltd. 25 - 47. Yang H, Reinach PS, Koniarek JP, Wang Z, Iserovich P and Fischbarg J. 2000. Fluid transport by cultured corneal epithelial cell layers. Br. J. Ophthalmol. 84(2): 199 - 204. Yue BY, Lin CC, Fei PF and Tso Mo 1984. Effects of chondroitin

sulfate

metabolism

trabecular

meshwork. Exp. Eye. Res. 38 (1): 35 - 44.

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