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Original Research
Recent biophysical characteristics of domestic water

sources in Owerri Metropolis, Nigeria.
Journal of Research in Biology
Authors: ABSTRACT:
Nwachukwu MI1*,
Eziuzor SC2, Duru MKC3,
Nwachukwu IO1,
Ukaga CN4, Udujih OS1 and The recent biophysical characteristics of domestic water sources in Owerri
Udujih GO5. 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.
2000, Owerri, Nigeria. (TCC), total Salmonella-Shigella count (TSSC), and total fungal count (TFC), as against
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.
7021, Aba, Nigeria. standards for domestic use. The high percentage occurrence of Salmonella species
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.
2000, Uturu, Nigeria. routinely reported in the clinics within the metropolis. With these findings, there is
need for public water supply authority within Owerri metropolis to improve in quality
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 Keywords:
Technology, P.M.B. 1526,
Bio-load, biophysical characteristics, infections, water sources, Owerri metropolis.
Owerri, Imo State, Nigeria.
Corresponding author: Article Citation:

Nwachukwu MI. 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 Accepted: 05 Aug 2013 Published: 11 Nov 2013
Web Address:
http://jresearchbiology.com/
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/
documents/RA0297.pdf. 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.
1066-1071 | JRB | 2013 | Vol 3 | No 6

An International
Scientific Research Journal www.jresearchbiology.com
Nwachukwu et al., 2013
INTRODUCTION microbiological and physiochemical characteristics of

Water of good quality is very important to health domestic water sources in Owerri metropolis Nigeria.
and man’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 of 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, “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.
treatment”. 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 as contamination may be ten-fold serially diluted before being plated out using the
intermittent. And considering the global data, morbidity spread plate technique in triplicates for total
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 were streaked, purified, and identified
aimed at providing recent information on the through biochemical, microscopic and macroscopic
1067 Journal of Research in Biology (2013) 3(6): 1066-1071
observations according to 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 fungal count. Rain water was next to borehole water in
sources include colour, taste, odour, pH. Iron, total 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 ≤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
RESULTS water sources are presented in figures 2 and 3,
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%),
2 3
1.5x10 to 1.5x10 cfu/ml. The total coliform count Klebsiella (4.35%) and Enterobacter (4.35%) as isolated
2
(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
2.5x10 cfu/ml, the total Vibrio count (TVC) ranged and the ties of Cryptococcus (14.28%) and
2
from 2.5 to 7.2x10 cfu/ml, and total fungal count (TFC) Saccharomyces (14.28%) as lowest.
ranged from 2.5 to 4.0x10 cfu/ml. The findings as shown Statistical analysis revealed a significant
in figure 1, make borehole water the best among the difference at ≤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.
highest bio-load in the present study. This makes it the
Cell Density (cfu/mi)
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

25
Percentage occurence (%)
20
15
10
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 Water sources Tolerance

A B C D E WHO
Colour (TCU) ( Units) colour less colour less dull colour less colour less 500
Odour no no ob ob no no
Taste no ob ob ob No no
pH 6.7 6.92 6.86 6.92 6.82 7.0 - 8.50
Conductivity (µs/cm) 146.2 23.6 45.5 28.4 3.4 500
Turbidity ( NTU) 1.0 20.37 7.77 00.0 1.5 50
Alkalinity (mg/ l) 0.0 2.00 5.00 04.0 1.0 600
Chlorine (mg/l) 0.0 0.00 0.00 00.0 0.0 200
Total Iron (mg/ l) ≤0.1 ≤0.1 ≤0.1 ≤0.1 ≤0.1 0.1
BOD (mg/l) 1.3 1.38 1.48 01.2 1.3 2.0
COD (mg/l) 121.45 137.18 137.18 120.2 117.58 196
TDS (mg/l) 0.2 11.7 11.7 0.1 0.1 -
TCU-true colour unit, no-not objectionable, ob-objectionable, NTU-nephlometric turbidity units.
A-borehole, B-Otamiri, C-Nworie, D-tap water, E-rainwater

45
Percentage occurence (%) 40
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

water supply by public water supply authority, the Nwachukwu CI and Otokunefor TV. 2006.
findings of the present study have also shown that the Bacteriological quality of drinking water supplies in the
University of Port Harcourt, Nigeria. Nigerian Journal of
improvement should as well include the quality of water
Microbiology, 20(3): 1383 – 1388.
distributed. Efficient distribution of portable water by
public water supply authority used to be the pride of the Onyeagba A, Ugbogu OC, Kanu IJ and Ogbu O.
2004. Laboratory Guide for Microbiology. Crystal
metropolis in the past.
Publishers, Owerri, Nigeria.
Raymond F. 1992. Problems of Water Supplies. EB and

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Sons Ltd UK. 123 – 126.
Antonine JPD and Dante C. 2008. Chemical levels in
drinking water. Applied Environmental Microbiology, 66 Tebutt THY. 1983. Principles of Quality Control.
(6): 2520 – 2525. Pergamon publishers, England.
American Public Health Association (APHA). 1998. UNICEF. 2004. Water. Environment and Sanitation.
Standard Methods for the Examination of Water and World Water Day 2004. Available online at
Wastewater. 20th ed. Washington, DC. www.unicef.org//wes/index.html.
Chukwura EI. 2001. Aquatic Microbiology. Octoba USEPA. 2001. Current Drinking Water Standards.
Press, Onitsha, Nigeria. 67 – 77. United States Environmental Protection Agency,
Washington, USA.
Cappucinno James G and Sherman W. 1992.
Microbiology: A Laboratory Manual. 3rd ed. Benjamin WHO. 2003. Water Supply, Sanitation and Hygiene
Cummings, California. 25 – 30. Development. Water. Sanitation and Health WHO,
Geneva.
Edema MO, Omemu AM and Fapetu OM. 2001.
Microbiology and physico-chemical analysis of different WHO. 1984. Guidelines for Drinking Quality. Drinking
sources of drinking water in Abeokuta, Nigeria. Nigerian Water Quality Control in Small Community Supplies,
Journal of Microbiology, 15(1): 57 – 61.
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

Submit your articles online at www.jresearchbiology.com
Determinative Bacteriology, 9th ed. Williams and
Wilkins Co., Baltimore. Advantages
Easy online submission
Ihekoronye AI and Ngoddy PO. 1985. Integrated Food Complete Peer review
Sciences and Technology for the Tropics. Macmillan Affordable Charges
Education Ltd. London and Oxford.95 – 195. Quick processing
Extensive indexing
Lemo OO. 2002. Bacteriology Determination of Water You retain your copyright
with Long Term Storage. BS.c thesis, University of submit@jresearchbiology.com
Agriculture, Abeokuta UNAAB, Abeokuta. 40 – 41. www.jresearchbiology.com/Submit.php.

Original Research
Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae and

the moth, Philosamia ricini
Authors: ABSTRACT:
Bendang Ao* and

Sentimenla. 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
Institution: were extracted by biochemical method and the subsequent electrophoretic analysis
Department of Zoology,
revealed that they were similar to chondroitin 4 – sulfate. Moreover, chromatographic
School of Sciences,
Nagaland University, analysis revealed different sugar components in the eyes of the two insects. It is
Lumami - 798627, concluded that acid mucopolysaccharides have structural and other physiological roles
Nagaland, India in the visual apparatus but no part in light and dark or photoperiodic adaptations.
Corresponding author: Keywords:

Bendang Ao. Mucopolysaccharides, Rhabdome.
Email: Article Citation:

Bendang Ao and Sentimenla.
Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae and the moth,
Philosamia ricini.
Dates:
Web Address:
http://jresearchbiology.com/ Received: 13 Mar 2013 Accepted: 21 Sep 2013 Published: 11 Nov 2013
documents/RA0340.pdf.
licenses/by/2.0), which gives permission for unrestricted use, non-commercial, distribution and
1072-1085 | JRB | 2013 | Vol 3 | No 6

An International Scientific
Research Journal www.jresearchbiology.com
Bendang, 2013
INTRODUCTION units are usually N-acetylated and sometimes N-sulfated,

Kennedy and White (1983) introduced the term while the acidic units are sometimes O-sulfated
“mucopolysaccharides” to describe 2-amino-2- (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 within the mucopolysaccharides category
protein derivative. They can be those that contain uronic irrespective of the ways in which the term has been used,
acid and those that are neutral. 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.
sulphate and hyaluronic acid respectively. These terms Presence of acid mucopolysaccharides in the
i . e. , AMPs an d SMPs (sul pha t ed a ci d 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
Meyer (1938) 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; Fitzsimmons et al., 1992;
similar physico-chemical properties isolated from Takahashi et al., 1993; Goes et al., 1999; Kato et al.,
connective tissues. Later on, the terms 1999), in chick cornea (Conrad et al., 1977; Li et al.,
“gl yc osa m in ogl yca n s” “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 bacterial polysaccharides and 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).
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 et al., 1977), interstitial matrix
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) ha d indi ca t ed that and Robert, 1967) etc. In the case of insects, AMPs have
glycosaminoglycans may have specific 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 within the carbohydrate chain. Limulus polyphemus and Macrobrachium birmanicum
“Glycosaminoglycan” is the systematic name for the (Dey et al., 1978), Musca domestica, Apis cerana
carbohydrate residues which form linear chains of indica (Dey, 1980)
alternating acidic and basic monosaccharides. The basic
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 rpm and precipitate was collected and dried. The
compound eye of the insects viz., 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 evaporated to dryness. The dried residue was then
The eyes were separated from live insects and 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 water in the ratio of 4:1:1 (v/v) as solvent (Giri and
thick sections were cut by microtome. The section were Nigam, 1954).
stained with Toluidine blue and Alcian blue (Humason, The chromatogram was developed with silver-
1971) for detection of mucopolysaccharides. nitrate (0.1 ml of saturated solution in 20 ml of acetone)
and sodium hydroxide (0.5 gm of NaOH in 25 ml of
Biochemical study according to Dietrich et al., (1977).
rectified spirit) as suggested by Trevelyan et al., (1950).
Extraction:
The chromatogram was then washed in 6N ammonium
Fresh eyes (1gm) were defatted in cold acetone
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 This was according to the method as described
by Leitner and Kerby, (1954). Streaks of the acid
forming a layer at the surface, and incubated at 37°C for
mucopolysaccharide samples were applied on Whatman
24 hours. After incubation, pH of the mixture was
No.1 paper strips using 0.1M phosphate buffer (pH 6.6)
brought to 11 with Conc. NaOH and maintained for six
at 4v/cm for 8 hours. After removal from the

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 with several standard a ci d
times in H2O. The strips were then dried at room mucop ol ysa cch ar ides sh owed that th e
temperature. mucopolysaccharides extracted resemble chondroitin
OBSERVATIONS 4-sulfate (Figure 5 and 6; Table 3 and 4).
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., i n di c a t i n g th e pr e s e n c e of 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 i n di ca t e s the pr e s en c e of a ci d by the glycosaminoglycans.
mucopolysaccharides. (Fig 1) In the case of the visual apparatus, they play a
Lens cuticle of the moth, Philosamia ricini: central role in the physiological maintenance of
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 analysis of the acid play important roles in “water binding” 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 Philosamia ricini (Figure 3 and 4; They based this on the fact that the corneal stroma of
Table 1 and 2). most vertebrates, including mammals, birds and teleosts
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 of 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 of mucopolysaccharides in
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

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Table 1: Ascending paper chromatogram of sugar Table 2: Ascending Paper chromatogram of some
components of the butterfly, Pieris brassicae and the standard sugar components. (Solvent used is butanol,
moth, Philosamia ricini. (Solvent used is butanol, acetic acid and water in the ratio of 4: 1:1 v/v)
acetic acid and water in the ratio of 4: 1:1 v/v)
Insect Rf value Identification Sugar Rf value
0.05 Lactose Raffinose 0.03
Butterfly,
Pieris brassicae
0.18 Galactose Lactose 0.05
0.33 Xylose Glucose 0.10
0.16 Galactose Sucrose 0.13
Moth, 0.33 Xylose Galactose 0.18
Philosamia ricini 0.43 Rhamnose Mannose 0.25
Fructose 0.28
errors of metabolism and/ or storage disorders (Matalon Xylose 0.34
Ribose 0.38
et al., 1974a; Hall et al., 1978; Neufeld and Fratantoni,
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 , insects.
and Ca++, and also with cationic groups of 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
++ +
preference for binding Ca 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
++
substances act as a store for Ca 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 the occurrence of 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 also suggested that stromal
cuticle where it performs some special functions transparency is correlated with the presence of normal
Bendang 2013
Fig. 3: Ascending paper chromatogram showing the Fig. 4: Ascending paper chromatogram showing the
sugar components of the mucopolysaccharides from sugar components of the mucopolysaccharides from
the eye of the butterfly, Pieris brassicae. the eye of the moth Philosamia ricini.
Fig. 5: Paper electrophorectic movement patterns Fig. 6: Paper electrophorectic movement pattern of
of the crude mucopolysaccharides from the eyes of the crude mucopolysaccharides from the eyes of the
the butterfly, Pieris brassicae. 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;

Bendang, 2013
Table 3: Paper electrophorectic movement patterns of Table 4: Paper electrophorectic movement patterns of
the crude mucopolysaccharides from the eyes of the
some standard mucopolysaccharides. (Solvent used is
butterfly, Pieris brassicae and the moth, Philosamia
phosphate buffer of pH 6.5)
ricini. (Solvent used is phosphate buffer of pH 6.5)
Distancetravelled Acid mucopolysaccharide Standard Distance travelled

Insect
(cms) type mucopolysaccharides (cms)
Butterfly, Heparin 5.5
6.4 Chondroitin 4-sulfate Chondroitin 4-sulfate 6.6
Pieris brassicae
Moth,
6.8 Chondroitin 4-sulfate
Heparan sulfate 7.2
Philosamia ricini Chondroitin 6-sulfate 7.6
Keratan sulfate 8.7
Bettelheim and Plessy 1975; Castoro et al., 1988 and Dermatan sulfate 10.0
Cristol et al., 1992) and this was thought to be related to (Moozar and Moozar, 1972).
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 it is the high negative charge of 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.
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, 1969). The protein polysaccharide noted that they are present not only in the corneal lens
complex provides a more stable and 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 that during corneal development of
Bendang, 2013
vertebrates, rise in transparency of stroma was found to the polysaccharides for the normal function of the
be associated with occurrence of mucopolysaccharides corneal stroma. Acta. Ophthal. 48(3): 455 - 464.
(Anseth and Fransson, 1970). Balazs EA. 1965. Amino sugar-containing
It is thus concluded that AMPs do indeed play macromolecules in tissues of the eye and ear. In: The
various roles in the physiology of vision, but no amino sugars. Balazs EA and Jeanloz RW, Eds. Vol 2A,
401 - 460. Academic press, New York.
photoperiodic adaptational mechanisms can be attributed
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

The present investigation revealed that mucopolysaccharides of cattle retina. Biochem. J. 108
(part 1): 75 - 88.
mucopolysaccharides are present in the ocular tissues
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(glycosaminoglycans) of the retina. Identification,
the insects studied i.e., Pieris brassicae, and
distribution and possible biological role. Biol.
Philosamia ricini. Moreover, the analysis of sugar
Ophthalmol. 79: 5 - 31.
components show that the ocular tissues of both the
Bettelheim FA and Plessy B. 1975. The hydration of
insects have similar sugars – galactose and xylose,
proteoglycans of bovine cornea. Biochem. Biophys. Acta.
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(in Pieris brassicae) and rhamnose (in Philosamia
Blochberger TC, Vergnes JP, Hempel J and Hassell
ricini), but no definitive conclusion can be drawn on the
JR. 1992. cDNA to chick lumican (Corneal keratan
matter of this difference pending further studies. It is sulfate proteoglycan) reveals homology to the small
thus concluded that acid mucopolysaccharides have interstitial proteoglycan gene family and expression in
structural and other physiological roles in the visual muscle and intestine. J. Biol. Chem. 267 (1): 347 – 352.
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Original Research
Diversity of Wetland dependent birds around the Bhadra Reservoir

Project (BRP) area, Karnataka
Authors: ABSTRACT:
Dayananda GY.
The study of bird species inhabiting certain wetlands around Bhadra
Reservoir Project (BRP), Shivamogga, Karnataka was carried out from February 2008
Institution: to January 2010. The total of 68 species of wetland birds belonging to nineteen
Department of P.G. Studies families and six orders. Of these, Anatidae (15%) and Ardidae (13%) have more than
and Research in Applied nine species. The diversity may be attributed the moderate volume of water storage,
Zoology, Bioscience availability of food and assured protection to these birds. Additionally we recorded
Complex, Jnana Sahyadri, seven types of migratory birds visiting these ponds. Those include White-necked Stork,
Kuvempu University, Shoveler, Pintail, Grey Plover, Curlew, Ringtailed-fishing Eagle and Black-winged Stilt.
Shankaraghatta – 577 451. All these wetlands are important places for foraging activity of wetland birds. In order
Shimoga. to protect these wetland birds, the wetlands should be conserved by controlling
encroachment, pollution and other anthropogenic activities.

Dayananda GY. 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.
Web Address: Dates:

http://jresearchbiology.com/ Received: 06 July 2013 Accepted: 22 July 2013 Published: 04 Sep 2013
1054-1059 | JRB | 2013 | Vol 3 | No 6

An International
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 (2.94) = Ciconiidae (2.94) =
method (Colin et al., 1993; William, 1997). The Scolopacidae (2.94 ) = Laridae (2.94 ) = Alaudidae
residential status and abundance criteria was calculated (2.94) = Corvidae (2.94) = Ploceidae (2.94)
using presence and absence scoring method and then >Podicipedidae (1.47) = Recurvirostridae (1.47) =
Dayananda, 2013
Table 1. Wetland bird diversity around the Bhadra Reservoir Project Area
Sl.
Order Family Common Name Scientific Name RS AS FH
No
Podicipediformes Podicipedidae Little Grebe Tachybaptus ruficollis R C P
Pelecaniformes Phalacrocoracidae Little Cormorant Phalacrocorax niger RM VC P
Oriental Darter Anhinga melanogaster RM UC P

Ciconiiformes Ardeidae Grey Heron Ardea cinerea RM C P
Purple Heron Ardea purpurea RM C P
Pond Heron Ardeola grayii R VC P
Night Heron Nycticorax nycticorax R UC P
Cattle Egret Bubulcus ibis R VC P
Large Egret Casmerodius albus RM VC P
Median Egret Mesophoyex intermedia R VC P
Little Egret Egretta garzetta R VC P
Chestnut Bittern Ixobrychus cinnamomeus RM UC P
Threskiornithidae Black-headed Ibis Threskiornis melanocephalus R VC P
Black Ibis Pseudibis papillosa RM C P

Eurasian Spoonbill Platalea leucorodia RM R P
Glossy Ibis Plegadis falcinellus RM C P
Ciconiidae White-necked Stork Ciconia nigra M R P
Open-bill Stork Anastomus oscitans R UC P
Anseriformes Anatidae Lesser-whistling Teal Dendrocygna javanica R C O
Common Teal Anas crecca RM C O
Spot-billed Duck Anas poecilorhyncha RM VC O
Garganey Anas querquedula R UC O
Nakta or Comb Duck Sarkidiornis melanotos R UC O
Shoveler Anas clypeata M R O
Cotton Teal Nettapus coromandelianus R VC O
Mallard Anas platyrhynchos RM UC O

Pintail Anus acuta M R O
Brahminy Duck Tadorna ferruginea RM UC O
Accipitridae Common Pariah Kite Milvus migrans R VC C
Brahminy Kite Haliastur indus R VC C
Ring tailed fishing Eagle Haliaeetus leucoryphus M R C
Gruiformes Rallidae White-breasted Water Amaurornis phoenicurus R VC I,G

hen
Indian Moorhen Gallinula chloropus R VC O
Purple Moorhen Porphyrio porphyrio R VC O
Common Coot Fulica atra R VC O

Dayananda, 2013
Charadriiformes Jacanidae Bronze-winged Jacana Metopidius indicus R VC I/G

Hydrophasianus chirurgus RM VC I/G
Pheasant-tailed Jacana
Brown Crake Amaurornis akool R C I
Water Cock or Kora Gallicrex cinerea LM C I
Charadriidae Red-wattled Lapwing Vanellus indicus R VC I
Yellow-wattled Lapwing Vanellus malabaricus R VC I
Little-ringed Plover Charadrius dubius RM C I

Grey Plover Pluvialis squatarola M R I
Curlew Numenius arquata M R I
Common Sandpiper Actitis hypoleucos RM VC I
Marsh Sandpiper Tringa stagnatilis R C I

Recurvirostridae Black-winged Stilt Himantopus himantopus M R I
Scolopacidae Painted Snipe Rostratula benghalensis R C I
Common Snipe Gallinago gallinago RM C I
Laridae Indian River Tern Sterna aurantia R C P
Common Tern Sterna hirundo RM C P

Alcedinidae Lesser-pied Kingfisher Ceryle rudis R C P
Small-blue Kingfisher Alcedo atthis R C P
Blue-eared Kingfisher Alcedo meninting R C P

White-breasted King- Halcyon smyrnensis R VC P
fisher
Stork-billed Kingfisher Pelargopsis capensis R C P
Alaudidae Crested Lark Galerida cristata R C I
Black-bellied Finchlark Eremopterix griseus R UC I
Sturnidae Indian Myna Acridotheres tristis R VC I

Corvidae House Crow Corvus splendens R VC O
Jungle Crow Corvus macrorhynchos R VC O
Motacillidae Large pied Wagtail Motacilla maderaspatensis R C I
White Wagtail Motacilla alba RM VC I

Yellow Wagtail Motacilla flava R C I
Yellow-headed Wagtail Motacilla citreola RM C I
Paddy Field Pipit Anthus novaeseelandiae R VC I

Ploceidae Baya weaver bird Ploceus philippinus R VC I
Black breasted weaver Ploceus benghalensis R VC I
bird
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.
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 REFERENCES
Podicipedidae, Recurvirostridae and Sturnidae had single Ali S. 1996. The Book of Indian Birds. BNHS, Oxford
University Press, Mumbai.
species each. Among the birds recorded in this study,
about 36.76 % (25 species) are both piscivores and
Ali S, Ripley SD and Dick JH. 1995. A Pictorial Guide
insectivores and 22.06 % (15 species) are omnivores and to the Birds of the Indian Subcontinent, Oxford
4.41 % (3 species) are carnivores respectively (Fig. 1). University Press, Delhi, 165.
In the present study, the analysis on the status
Anon. 1990. Wetlands of India - A Directory.
shows that twenty five species are common, twenty eight
Government of India, Forests and Environment
species very common, nine species uncommon and eight
Department, New Delhi.
species rare. The abundance of birds may be influenced
by availability food for birds in the form of plants, Bhatnagar SP, Shukla SK and Bhatnagar M. 2008.
vertebrates and invertebrates, some of them feed in An annotated checklist of waterfowl of the Didwana
inland saline lake, Nagaur, Rajasthan, India, Newsl. For
wetland soil, water column and dry landscape in and

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Bird Watchers, 48:53-55. Ahmadpour and Younes Yaghobzadeh. 2011. A three

years study of the diversity and density of waterfowl and
Colin J Bibby, Niel D. Burgess and David A Hill. waders in Sorkhrud International Wetland (October 2007
1993. Birds census Techniques. Academic press Ltd. –March 2010). Scientific Research and Essays,
London. 6(30):6317-6324.
Dayananda GY. 2008. Status of waterbirds at Nanda KNV, Sailaja K, Nagarjuna A. 2010. Avian
Ramanakere, Cheelur, Davanagere District, Karnataka. biodiversity indices and comparative chronobiology of
J.Aqua.Biol., 23(2): 32-35. Uppalapadu and Nelapattu bird protected areas of
Andhra Pradesh, India. J. Zool., 5(3):148-152.
Dayananda GY. 2009. Avifaunal diversity of Gudavi
Bird Sanctuary, Sorab, Shimoga, Karnataka. Our Nature, Rajashekara S and Venkatesha MG. 2011.
7:100-109. Community composition of aquatic birds in lakes of
Bangalore, India. J. of Env. Biol., 32(1):77-83.
Fleming RL(Sr), Fleming RL(Jr) and Bangdel LS.
2000. Birds of Nepal with reference to Kashmir and Rajpar MN and Zakaria M. 2010. Density and
Sikkim. First Adarsh Impression, Gaurav Offset, Delhi. diversity of water bird and terrestrial bird at Paya Indah
wetland reserve, Selangor Peninsular Malaysia. J. Biol.
Grimmett R, Inskipp C and Inskipp T. 2001. Pocket Sci., 10(7):658-666.
Guide to the Birds of the Indian Subcontinent. Oxford
University Press, New Delhi. Sonobe K and Usui S. 1993. A Field Guide to the
Water Birds of Asia. Wild Bird Society of Japan, Tokyo.
Huston MA. 1994. biological diversity, Cambridge
University Press, Cambridge. 3rd Edition.,78. William J Sutherland. 1997. Ecological Census
Techniques a handbook. Cambridge University Press
Inskipp C and Inskipp T. 1991. A Guide to the birds of U.K.
Nepal. Christopher Helm. London.
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 Advantages
subcontinent. Buceros, 6(1) : 1-37.
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Karimi, Seyed Mahmoud Ghasempouri, Mousa

Original Research
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
Animal Science/Fisheries, of the maggots ranged from 55.4% in maggots grown on pig dung to 57.42% in
Abia State University, maggots grown on cattle blood. The crude fibre contents ranged between 0.32% and
Umuahia Campus. Abia 0.21%. Maggots produced from pig dung and cattle blood recorded the highest ash
State, Nigeria. content and the values were 11.09% and 11.20% respectively. Moisture content was
highest (10.14%) for maggots produced from cattle dung. Fat content of the maggots
2. Department of Nutrition produced from the different livestock wastes ranged between 21.06% and 22.66%.
and Dietetics Imo State Significant differences (p<0.05) in the proximate composition of the maggots were
University, P. M. B. 2000, only observed in the crude fiber, ash and moisture contents. The results from this
Owerri, Imo State, Nigeria. study showed that the substrates used can produce substantial quantities of maggots
with varying degrees of success.

Afamdi Anene. maggots, proximate quality, livestock wastes
Email Id: 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.
Web Address: Dates:

http://jresearchbiology.com/ Received: 07 Aug 2013 Accepted: 12 Aug 2013 Published: 18 Oct 2013

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

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

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.
Anene et al., 2013
Data Collection, Sample and Data Analysis Table 1: Weight of maggots produced from
different livestock wastes
Maggots from each replicate were weighed to the
nearest 0.1g when wet and then weighed after drying to a
Mean Yield (g)
Treatments
constant weight at 35oC in an oven using a digital per Kg (N=3)
weighing balance. Dried maggots from each treatment Wet Weight Dry Weight
were blended into a smooth paste in a 3.8 L kitchen-type Poultry Droppings 58.730±0.34a 12.79±0.22a
Pig dung 08.180±0.22d 02.97±0.17d
blender (Warning Products, New Hartford, CT) which
Cattle dung 12.920±0.16c 04.18±0.52c
was thoroughly cleaned and dried between samples. Cattle Blood 21.770±0.31b 07.79±0.41b
Triplicate determination was made for each treatment. Means in the same column with different superscripts
Samples were analysed for crude protein (CP), crude 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 The results obtained in this study compared
using Duncan’s Multiple Range Test (Duncan, 1955). 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 maggots. These report however
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.
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

Anene et al., 2013
Table 2: Proximate composition of maggots produced from different livestock wastes

Treatments
Parameters Poultry droppings Pig dung Cattle Dung Cattle Blood
Crude Protein 56.25±0.21a 55.54± 0.15a 56.00a± 0.00 57.42a±0.00
Crude fibre 00.32±0.08a 00.26±0.05ab 00.21± 0.01b 00.29±0.06ab
Ash 10.80±0.17b 11.09±0.15a 10.90± 0.12b 11.20±0.11a
Ether Extract 22.32±0.09a 22.64±0.07 a
22.66± 0.21a 21.06±0.19a
Nitrogen Free Extract 00.17±0.04a 00.61±0.07a 00.07± 0.01a 00.16±0.07a
Moisture content 10.12±0.11b 09.84±0.12b 10.14± 0.21a 09.86±0.16b
* 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 content is an indication of the mineral content of feed
maggots produced from various substrates. This 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
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 2010.
contents, the ash, nitrogen free extract and moisture
AOAC. 1995. Official Methods of analysis AOAC,
composition were affected by the type of substrate used
International, P. Cunniff Sixteenth edition, Vol. II
in the study. The protein content in the maggots
Chapter 49, 1-49. Arlington, Virginia, United States.
produced from poultry (layer) droppings, cattle dung, pig
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
Calvert CC, Martins RD and Eby HJ. 1971. Housefly
production bringing about economic affordability of the
pupae as food for poultry. Journal of Economic
much needed animal protein. The results also show that
Entomology. 62(1): 939.
maggot meal is not a good source of fiber. This study
also further strengthens the role of maggots in Cao JunMing, Yan Jing, Huang YanHua, Wang
biodegradation of livestock/animal wastes and its GuoXia, Zhang RongBin, Chen XiaoYing, Wen
importance in the management of wastes in the industry. YuanHong, Zhou TingTing. 2012. Effects of
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 performance, antioxidant and non-specific
facilitate its incorporation into animal and fish feeds. immune indexes of juvenile Litopenaeus vannamei.
J. Fish. China, 36 (4): 529-537.
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Journal of Research in Biology An International Scientific Research Journal

Recent biophysical characteristics of domestic water

Corresponding author: Article Citation:

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

INTRODUCTION microbiological and physiochemical characteristics of

Journal of Research in Biology (2013) 3(6): 1066-1071 1068

Table 1. Physicochemical characteristics of water sources in Owerri metropolis

Parameters Water sources Tolerance

1069 Journal of Research in Biology (2013) 3(6): 1066-1071

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

Journal of Research in Biology (2013) 3(6): 1066-1071 1070

Raymond F. 1992. Problems of Water Supplies. EB and

Gerhardt P. 1994. Methods for General and Molecular

Holt JG, Bergey DH (ed.). 1994. Bergey’s Manual of

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

Acid mucopolysaccharides in the eyes of the butterfly, Pieris brassicae and

Bendang Ao* and

Corresponding author: Keywords:

Email: Article Citation:

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

INTRODUCTION units are usually N-acetylated and sometimes N-sulfated,

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

Journal of Research in Biology (2013) 3(6): 1072-1085 1074

Journal of Research in Biology (2013) 3(6): 1072-1085 1076

Journal of Research in Biology (2013) 3(6): 1072-1085 1078

Distancetravelled Acid mucopolysaccharide Standard Distance travelled

CONCLUSIONS Berman ER and Bach G. 1968. The acid

Journal of Research in Biology (2013) 3(6): 1072-1085 1080

1081 Journal of Research in Biology (2013) 3(6): 1072-1085

factor-mediated phosphorylation of PDGF-Rbeta in Kato T, Nakayasu K, Ikegami K, Obara T,

Journal of Research in Biology (2013) 3(6): 1072-1085 1082

1083 Journal of Research in Biology (2013) 3(6): 1072-1085

Journal of Research in Biology (2013) 3(6): 1072-1085 1084

Wang L, Fuster M, Sriramarao P and Esko JD. 2005.

Wigglesworth VB. 1965. The principle of Insect

Yang H, Reinach PS, Koniarek JP, Wang Z,

Yue BY, Lin CC, Fei PF and Tso Mo 1984. Effects of

Submit your articles online at www.jresearchbiology.com

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

Diversity of Wetland dependent birds around the Bhadra Reservoir

Corresponding author: Keywords:

Email Id: Article Citation:

Web Address: Dates:

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

INTRODUCTION percentage of birds occurrence was calculated to

Pelecaniformes Phalacrocoracidae Little Cormorant Phalacrocorax niger RM VC P

Oriental Darter Anhinga melanogaster RM UC P

Black Ibis Pseudibis papillosa RM C P

Mallard Anas platyrhynchos RM UC O

Gruiformes Rallidae White-breasted Water Amaurornis phoenicurus R VC I,G

Journal of Research in Biology (2013) 3(6): 1054-1059 1056

Charadriiformes Jacanidae Bronze-winged Jacana Metopidius indicus R VC I/G

Little-ringed Plover Charadrius dubius RM C I

Common Sandpiper Actitis hypoleucos RM VC I

Marsh Sandpiper Tringa stagnatilis R C I

Common Tern Sterna hirundo RM C P

Blue-eared Kingfisher Alcedo meninting R C P

Sturnidae Indian Myna Acridotheres tristis R VC I

White Wagtail Motacilla alba RM VC I

Paddy Field Pipit Anthus novaeseelandiae R VC I

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

Journal of Research in Biology (2013) 3(6): 1054-1059 1058

Bird Watchers, 48:53-55. Ahmadpour and Younes Yaghobzadeh. 2011. A three