Journal of Research in Biology
Original Research
Impact of the residue of Deltamethrin and Endosulfan pesticides on biochemical toxicity
and some neurotransmitter contents in different brain areas of male Albino mice
Authors:
Journal of Research in Biology
Somaya M. Ismail1,
Azza A. Said2,
Samira M. El-Sayad2.
Institution:
1. Zoology department,
Faculty of Science,
Cairo university.
2. Zoology department,
Faculty of Science,
Fayoum university.
Corresponding author:
Somaya M. Ismail
Email:
mragaa11@yahoo.com
Tel:
+201118244212
Web Address:
http://jresearchbiology.com/
documents/RA0324.pdf.
Journal of Research in Biology
An International Scientific
Research Journal
An International Scientific Research Journal
ABSTRACT:
Evaluating the action of the residues of pesticides on non-target organisms
has been of interest to many researchers. The present study aimed to evaluate the
pesticides deltamethrin and endosulfan on biochemical toxicity and some
neurotransmitter contents in different brain areas of male albino mice. The results
showed that the daily oral administration of deltamethrin and endosulfan caused a
significant decrease in neurotransmitter contents (NE, DA and GABA) in most of the
tested brain areas (cerebellum, striatum, cerebral cortex, hypothalamus, brain stem
and hippocampus). On the other hand a gradual significant reduction, ALT, AST and
ALP enzyme activities, while the glucose level and acid phosphatase increase were
observed in serum of mice treated with deltamethrin and endosulfan for two weeks.
Also, this study has a significant inhibition in the activities of enzymes in liver tissues of
treated mice including glutathione reductase. Meanwhile, the activity of lipid
peroxide, glycolytic (PK, PFK and GPI) and gluconeogenic enzyme activities
(F-1, 6-D-Pase) were significantly increased in liver tissues of treated mice in response
to treatment. Additionally, total protein and glycogen content showed a significant
reduction in liver tissues of mice treated with deltamethrin and endosulfan for two
weeks. It was concluded that the pollution of the aquatic environment by
deltamethrin and endosulfan pesticides, would adversely affect the metabolism of the
mice.
Keywords:
Deltamethrin, Endosulfan pesticide, Laboratory-bred strain Swiss albino male
mice, neurotransmitter contents (NE, DA and GABA).
Article Citation:
Somaya M. Ismail, Azza A. Said and Samira M. El-Sayad.
Impact of the residue of Deltamethrin and Endosulfan pesticides on biochemical toxicity
and some neurotransmitter contents in different brain areas of male Albino mice.
Journal of Research in Biology (2013) 3(4): 954-966
Dates:
Received: 12 Jan 2013 Accepted: 22 Feb 2013 Published: 24 May 2013
This article is governed by the Creative Commons Attribution License (http://creativecommons.org/
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954-966 | JRB | 2013 | Vol 3 | No 4
www.jresearchbiology.com

INTRODUCTION
Using pesticides is an important procedure for
enhancing agriculture yield. However, the great
consciousness, brought back upon their deleterious
effects on human, animal and environmental health,
leading to the shortage of their use by imposing various
rules (Ahmsd et al., 2010; Botella et al., 2004).
Among pesticides, Deltamethrin, which is a type
II pyrethroids, has a wide acceptability, and is used in
agriculture and forestry because of its high
activity against a broad spectrum of insect pests
(Villarini et al., 1998). The oral route constitutes the
main sources of general population exposure to this
pesticide which is ingested within food and water
(Barlow et al., 2001).
It has been reported that deltamethrin caused
an oxidative damage in liver and intestine of
Carassius auratus gibelio explained by an increase of
LPO level and an enhancement of antioxidative defence
parameters (Dinu et al., 2010). Oral absorption of
deltamethrin is rapid and is metabolized with microsomal
enzyme system in liver and with tissue esterase present
in intestinal wall and liver in addition to plasma
carboxylesterases (Usmani et al., 2006). According
to Simsek et al., (2008), Deltamethrin applied at different
concentrations of 25, 50, 100, 200, 400, 800 and
1600 mg /L, for 1,24, 48, 72 and 96 h increased lipid
peroxidation which is accompanied by a decrease of
reduced glutathione and catalase activity in digestive
gland and gill of fresh water mussel.
Endosulfan (6, 7, 8, 9, 10-hexachloro -1,5,5a,6,
9a-hexahydro-6,9-methano -2, 4, 3 benzodioxathiepine-3
-oxide) is a broad-spectrum organochlorine pesticide
(insecticide and acaricide) first registered for use in the
United States in 1954 to control agricultural insect and
mite pests on a variety of fruits, vegetables, rice, grains,
tea, coffee, cotton and also in animal farm and houses
(US EPA). Results from a global monitoring network for
persistent organic pollutants revealed that endosulfan is
955
Ismail et al., 2013
abundant in the environment and its use is increasing
(Pozo et al., 2006; Harner et al., 2006). It reaches aquatic
systems through direct application, as well as spray drift
and runoff from agricultural areas (Broomhall, 2002;
Jergentz et al., 2004 and Rand et al., 2010).
It is known that exposure to pesticides during
development may interfere with the normal development
of neurotransmitter systems and cause their direct
damage (Richardson et al., 2006). The central nervous
system (CNS) during development is particularly
susceptible to the toxic effects of xenobiotics (Tilson,
2000). The mechanism by which these effects occur is
not known but currently it is assumed that the
monoaminergic neurotransmitters play a role during
development, defined as morphogenetic (Buznikov
et al., 1996; Levitt et al., 1997; Nicotra and Schatten,
1990).
Organophosphate pesticide represent one of the
worlds most commonly used agrochemical.
Consequently, many of its residues are frequently found
in the environment. The aim of this study was to
determine the effects of the pesticides, deltamethrin and
endosulfan on biochemical toxicity and some
neurotransmitter contents in different brain areas of male
albino mice.
MATERIALS AND METHODS
Pesticides
Deltamethrin
Deltamethrin is a synthetic pyrethroid pesticide
[ ( S ) -a c ya n o-3 - ph en ox ybe n z yl ( 1 R, 3 R) - 3 -( 2, 2-
dibromovinyl)-2,2-dimethylcyclopropanecarboxylate]
with molecular formula (C22H19 Br2NO3). Solubility in
water is <0.1 mg/L at 25oC. Relative molecular mass of
the compound is 505.2 g/mol, and the melting point is
100oC (Figure 1).
Endosulfan
Endosulfan is an off-patent organochlorine
pesticide and acaricide that is being phased out globally
Journal of Research in Biology (2013) 3(4): 954-966
Ismail et al., 2013
[6, 7, 8, 9, 10, 10-Hexachloro-1, 5,5a, 6, 9, 9a-hexahydro
-6, 9-methano-2,4,3-benzodioxathiepine-3-oxide]. With
the molecular formula of (C9H6Cl6O3S). Solubility in
water is 0.33 mg/L. Relative molecular mass is found to
1
be 406.93 g mol ,and the melting point is 70-100C,
343-373 K, 158-212 F (Figure 1)
Fig. 1. Chemical structure of the pesticides
Deltamethrin and Endosulfan
Fig. 2. Changes (%) of activities of glucose level (GL),
some enzymes (alanine aminotransferase (ALT),
aspartate aminotransferase (AST), Alkaline
phosphatase (ALP), acid phosphatase (ACP) in serum
of male mice,. Lipid peroxide (LP) glutathione
(GSH) , pyruvate kinase (PK), phosphofructokinase
(PFK), glucose phosphate isomerase (GPI), Fructose -
1, 6-diphosphatase (F-1, 6-ase) enzymes, Total protein
(TP), glycogen content in tissues of male mice liver
exposed to LC25 of Deltamethrin and Endosulfan
pesticides for 2 weeks.
Journal of Research in Biology (2013) 3(4): 954-966
Animals
Swiss albino male mice of 10 weeks old with an
average weight of 28.52.5 g obtained from the National
Research Centre, Cairo, Egypt were used. They were
maintained in a well ventilated animal house. They were
housed in large polypropylene cages with free access to
food and water ad labium during the course of
the experiment. Animals were housed in groups
(5 animals/ group) and maintained under standard
conditions of temperature (23C to 25C), a relative
humidity of 65% to 86% and in a schedule of 12 hours of
light and 12 hours of dark.
Animal treatment
The animals were divided into three groups (n=6)
of equal number, The control group (1) was orally and
daily administered with equivalent amount of the vehicle
(distilled water) for two weeks, the second group was
given drinking water with 1.28 mg/kg BW of
deltamethrin (Yousef et al., 2006) during two weeks of
oral and daily administration and the third group was
orally and daily administered with endosulfan
(1.5 mg /kg BW). At the ends of the experimental period
(2 weeks), the mice were sacrificed under diethyl ether
anesthesia at fasting state.
Effect of deltamethrin and endosulfan (pesticide) on
some neurotransmitter contents in different brain
areas of male albino mice
During the experiment six mice of each group
were decapitated each week till the end of the 2-week
duration times. The mice were killed by sudden
decapitation at the designed times. The brain was rapidly
and carefully excised and then dissected on dry ice glass
plate according to the method of Glowinski and Lversen
(1966) into the following regions; cerebellum, striatum
cerebral cortex, hypothalamus, brain stem and
hippocampus. Brain tissues were wiped dry with filter
paper, weighed, wrapped in plastic films and then in
aluminum foil and quickly frozen in dry ice. NE and DA
were extracted and estimated in the brain tissues
956
 Ismail et al., 2013

according to the method of Chang (1964) modified by
Ciarlone (1978). GABA were extracted and estimated in
the brain tissues according to the method of Sutton and
Simmonds (1973). The fluorescence was measured in
Jenway 6200 fluorometer.
Effect of deltamethrin and endosulfan (pesticide) on
biochemical toxicity of male albino mice
Serum samples were obtained by
centrifugation of blood of six rats of each group at
4000 rpm for 15 min at 4C, and were then divided in to
Eppendorf tubes. Isolated sera from each group were
stored at -20C until they were used for the analyses. For
preparation of tissue homogenates of mouse liver tissue
of six mice of each group, one gram of liver tissues of
mouse from each group was homogenized in 5 ml
distilled water at pH 7.5.A glass homogenizer was used
and the homogenate was centrifuged for 10 minutes at
3000 rpm, fresh supernatant was used.
The levels of serum alanine aminotransferase
(ALT), aspartate aminotransferase (AST) were measured
according to Reitman and Frankel (1957). Alkaline
phosphatase (ALP) was measured according to Belfield
and Goldberg (1971) and acid phosphatase (ACP) was
measured according to Wattiaux and De Duve (1956)
and sera glucose concentrations (GL) were determined
according to the glucose oxides method of Trinder
(1969). Total protein (TP) content was determined
according to Bradford (1976) Determination of tissues
glycogen was evaluated according to Nicholas et al.,
(1956). Lipid peroxide ( LP) was measured according to
Buege and Aust (1978). Glutathione (GSH) was
measured according to Moron et al., (1979) Pyruvate
kinase (PK) relative activity was measured
spectrophometrically by the method of Bucher and
the pfleiderer (1975). phosphofructokinase (PFK) was
measured according to Zammit et al., (1978) Glucose
phosphate isomerase (GPI) was measured according to
King (1965). Fructose -1, 6-diphosphatase (F- 1, 6-ase)
was measured according to Sand et al., (1980). All
biochemical l parameters determined in this study were
determined spectrophotometrically, using reagent kits
purchased from BioMerieux Company, France. Kits
purchased from BioMerieux Company, France.
Statistical analysis
The results obtained in the present work are
represented as means standard deviation (SD), and
were analyzed using analysis of variance (ANOVA). The
significance of difference between means were
calculated using the Duncan Multiple Range Test (Steel
and Torrie, 1980).
RESULTS
Results in Table 1 showed that the daily oral
administration of deltamethrin and endosulfan resulted in

Table (1): Effect of oral administration of Deltamethrin and Endosulfan on dopamine (DA) content in the
different brain areas of male albino rat.
Cerebellum Striatum Cerebral cortex Hypothalamus Brain stem Hippocampus
Pesticides
mean S.E. mean S.E. mean S.E. mean S.E. mean S.E. mean S.E.

C 122.7 0.72 280.50.64 52.30.084 433 4.2 310.20.45 222.10.6

Deltamethrin T 82.31.2* 186.60.6* 36.50.21* 146.32.1** 254.51.4 160.20.62
% 32.79% 33.69% 30.21% 66.21% 17.69% -27.87%
C 122.7 0.72 280.50.64 52.30.084 4334.2 310.20.45 222.10.6

Endosulfan T 48.22.42** 145.122.3** 21.40.73** 116.42.5*** 192.63 1.5** 112.61.6**

% 60.72% 48.26% 59.1% 73.12% 38.11% 49.30%

- Statistical analyses were performed between control (C=6) and treated (T=6) animals by using paired t' test
% : Percentage of change from control *p< 0.05,**p< 0.01 & ***p< 0.001

957 Journal of Research in Biology (2013) 3(4): 954-966

Ismail et al., 2013

a significant decrease in DA content in all brain area.
The maximal decrease (p<0.001) in DA content was
found in the hypothalamus of mice treated with
deltamethrin and endosulfan at the concentrations of
66.21% and 73.1%, respectively. Also, Table 2 showed
that the daily oral administration of deltamethrin and
endosulfan caused a significant (p<0.001) decrease in
GABA content in all the brain area, the maximal
decrease (p< 0.001) in GABA content was found in
brain stem of mice treated with deltamethrin and
endosulfan at the concentration of 72.52% and 80.52%,
respectively.
The results obtained from Table 3 showed that
the maximal decrease (p<0.001) in NE content was
found in the hypothalamus of mice treated with
deltamethrin and endosulfan at the concentrations of
54.1% and 62.89%, respectively.
The results in Table 4 showed that a clear
reduction (P<0.001) in liver enzyme activities in serum
of mice treated with deltamethrin and endosulfan as
compared to the control mice. On the other hand, the
glucose concentration and Acid phosphatase in serum of
treated mice showed a marked increase (P<0.001) in
comparison with the control group. Glycogen content in
tissues of treated mice showed a significant (p>0.001)
decrease in comparison with the control group. The
reduction rates were 36.32% and 58.24% for mice
treated with deltamethrin and endosulfan, respectively
(Table 7).

Table (2):Effect of oral administration of Deltamethrin and Endosulfan on gama-butyric acid (GABA) content
in the different brain areas of male albino rat.
Cerebellum Striatum Cerebral cortex Hypothalamus Brain stem Hippocampus
Pesticide
mean S.E. mean S.E. mean S.E. mean S.E. mean S.E. mean S.E.

C 165.70.65 154.210.8 44.20.62 321.60.82 121.20.197 204.31.6

Deltamethrin T 102.61.3** 92.60.428** 35.20.8 29250.43* 33.30.764** 98.80.577**
% -61.97% 39.82% 18.1% 9% 72.52% 51.64%
C 165.70.65 154.210.8 44.20.62 321.60.82 121.20.197 204.31.6
Endosulfan T 60.61.2*** 72.40.87** 28.60.83*** 2521.6 23.60.82* 88.41.6***
% 63.43% 53.1% 35.29% 21.64% 80.52% 56.73%

- Statistical analyses were performed between control (C=6) and treated (T=6) animals by using paired t' test.
% : Percentage of change from control. *p< 0.05,**p< 0.01 & ***p< 0.001

Table (3): Effect of oral administration of Deltamethrin and Endosulfan on norepinephrine (NE) content
in the different brain areas of male albino rat.

Cerebellum Striatum Cerebral cortex Hypothalamus Brain stem Hippocampus

Pesticide
mean S.E. mean S.E. mean S.E. mean S.E. mean S.E. mean S.E.

C 102.61.4 434.21.6 64.61.54 462.22.11 3420.53 233.11.4

Deltamethrin T 77.50.56** 344.231.4** 35.20.54* 212.2052** 243.10.45** 106.20.62***
% 24.64% 20.72% 48.57% 54.1% 28.95% 45.44%
C 102.61.4 434.21.6 64.61.54 462.22.11 3420.53 233.11.4
Endosulfan T 48.20.62** 223.30.61** 22.31.4*** 210.81.1*** 168.51.4*** 86.50.83***
% 53% 48.57% 65.48% 99.45% 50.73% 62.89%
- Statistical analyses were performed between control (C=6) and treated (T=6) animals by using paired t' test.
% : Percentage of change from control. *p< 0.05,**p< 0.01 & ***p< 0.001

Journal of Research in Biology (2013) 3(4): 954-966 958

 Ismail et al., 2013

The present result in Table 5 indicated that a

% Change
significant increase in lipid peroxide accompanied with a

Data represent mean values of five replicates. Within columns for dose, time and (dose x time), mean values followed by different letters are statistically significantly
11.96%

37.27%
phosphatase (ACP)
significant reduction in glutathione and total protein in

Acid liver enzyme activities of mice treated with deltamethrin

and endosulfan as compared to the control mice.

8.84 F0.22
6.44 0.23

7.21 0.22
The present results in tables (6, 7) demonstrate a
significant elevated level of glycolytic (PK, PFK and
GPI) and gluconeogenic enzyme activities (F-1,6-D-
Table 4: Effect of Deltamethrin and Endosulfan on liver function enzymes in serum of male mice.

% Change

Pase) in tissue of mice treated with deltamethrin and

46.26%
11.8%
endosulfan as compared to the control. The elevation
ALP

rates in the activities of PK, PFK, GPI and F-1-6, D-Pase

3.61 0.03

2.85 0.05

1.94 0.01
enzymes were 97.36%, 76.1%, 74.84% and 69.1%,
respectively for mice treated with endosulfan.

DISCUSSION
% Change

18.48%

49.52%
(umole/mg protein/min.)
Liver function enzymes

Many monoamine neurotransmitters, including

different based on LSD at P = 0.05.

DA , NE and GABA are important in the regulation of

ALT

brain development prior to assuming their roles as

8.36 0.338

6.835 0.12

4.22 0.45

transmitters in the mature brain (Whitaker-Azmitia,

1992; Di Pino, 2004; Ansorge, 2008), thus any
circumstance that affects these neurotransmitters in the
developing brain can alter the final structure and function
% Change

44.87%

44.8%

of the brain. Developmental neurotoxicity involves

alterations in behavior, neurophysiology.
AST

From the present results, it is clear that the daily

18.21 0.432

14.21 0.316

10.04 0.311

oral administration of deltamethrin and endosulfan

caused reducing side effect in some neurotransmitter
tissue in the brain and a significant decrease in
% Change

neurotransmitter contents (NE, DA and GABA) in most

49.7%

64.9%

of the tested brain areas. Cerebellum which is

Glucose (GL)
mg/g tissue

responsible for the voluntary movement; pons and

medulla oblongata which is responsible of essential
44.5 0.64
22.4 1.5

15.6 1.8

reflexive acts; striatum which is a brain region

responsible for motor activity; cerebral cortex is
responsible for sensation including visual, auditory and
olfactory as well as motor coordination and association,
Deltamethrin

Endosulfan

also is responsible for higher mental function such as

Control

thinking, planning, reasoning, memory and

consciousness and hippocampus, this is the key area
959 Journal of Research in Biology (2013) 3(4): 954-966
Ismail et al., 2013
Table 5: Effect of Deltamethrin and Endosulfan on lipid peroxide, glutathione and total protein in male mice liver.
Lipid peroxide(LP) Glutathione(GR) Total protein(TP)
(ug/g tissue) (ug/g tissue) (mg/ml)

% change % Change % change

Control 0.65 0.01 30.22 1.22 52.44 2.11

Deltamethrin 0.95 0.57 -46.15% 20.20 1.12 33.16% 41.11 1.15 21.61%

Endosulfan 1.220.06 -87.69% 15.16 0.85 49.83% 31.22 1.65 40.46%

concerned with learning (Ansorge, 2008). Brain stem is
responsible for integration of coordination of essential
reflexive acts such as swallowing, vomiting and
respiration (Bloom, 1983).
Our findings support the idea that deltamethrin
and endosulfan is neurotoxic in the developing brain.
The present result found that these pesticides induced a
decrease in DA levels in cerebellum, striatum, cerebral
cortex, hypothalamus, brain stem and hippocampus of
treated mice. The loss of hippocampus DA levels was
higher in treated mice. DA is an important component of
the neuroendocrine mechanism that regulates the
activation of male sexual behavior in mammalian species
(Castagna and Ball, 1997). Moreover, steroidogenesis in
the brain may play a critical role in mammalian brain
developmental of both sexes (Konkle and McCarthy,
2011). Steroids play a role in the development of
catecholamines systems (Leret, 2009; Muneoka et al.,
2010; Pappas et al., 2010).
It is known that DA is the major compound
involved in the control of the motor system. Bernardi and
Palermo-Neto, (1983) showed that locomotion and
rearing frequencies observed in an open field might be
used to detect drug-induced dopaminergic interference.
Locomotors activity as measured in the open field
appears to be associated with the dopaminergic system
(Chiavegatto et al., 1998). Also, in the present study, we
similarly found a loss of the NE and gamma-butyric acid
(GABA) content in the cerebellum, striatum, cerebral
cortex, hypothalamus, brain stem and hippocampus. The
loss of brain stem DA levels and the loss of hippocampus
GABA levels were higher in treated mice.
These effects may represent a large number of
actions involved in the development of synaptic
dysfunction in these neurotransmitter systems that
ultimately contribute to behavioral anomalies.
Nevertheless further behavioral testing is needed to
confirm this suggestion. Moreover, the present findings
might indicate that prenatal and postnatal exposure to
pesticide altered the program for developmental of DA,
NE, and GABA synaptic functions. Given that, the
dysfunction in serotonin and dopamine systems is
involved such as appetite, affective, locomotion,
learning, neurological and neuropsychiatric disorders
(Insel et al., 1990; Kaye, 2008), further testing of this
function is needed to confirm that alteration of these
neurotransmitter systems is the cause of some of these
dysfunctions. In general, our results support the

Table 6: Effect of Deltamethrin and Endosulfan on some glycolytic enzymes in male mice liver.
Glycolytic enzmes (umole/mg protein/min.)
PK PFK GPI
% change % change % change
Control 4.16 0.26 7.44 1.16 77.34 2.43
Deltamethrin 7.18 1.44 -72.6% 10. 1 1.22 -35.75% 113.50 3.2 -46.81%
Endosulfan 8.23 1.64 -97.36% 13.1 1.23 -76.1% 135.22 6.4 -74.84%
Data represent mean values of five replicates. Within columns for dose, time and (dose x time), mean values followed by
different letters are statistically significantly different based on LSD at P = 0.05.

Journal of Research in Biology (2013) 3(4): 954-966 960


Table 7: Effect of Deltamethrin and Endosulfan on Glycogen and some Gluconeogenic enzymes in male mice liver.
Glycogen( mg/g tissue )
Control 6.8 0.64
Deltamethrin 4.330.64
Endosulfan 2.841.02
suggestion that at least some of the effects of these
disorders that are increasing in humans can be caused by
exposure to neurotoxin environmental contaminants
(Slikker W and Schwetz, 2003).
In conclusion, the results observed in this study
reinforce the idea of the use of neurochemical measures,
such as the DA, NE and GABA content and its
metabolites in brain regions as indicators
neurotoxicity, including developmental neurotoxicity,
induced by chemical agents. Because of serotonergic
dysfunction is involved in appetite and affective
disorders, and the catecholamine DA and NE have been
most often linked to the behavioral pathology of a
number of neurological and psychiatric disorders, studies
of pesticide on DA, NE- and GABA. Related behaviors
in animal models will be needed to clarify the outcomes
of long-term alterations in noradrenergic, serotonergic
and dopaminergic systems identified here.
Concerning, ALT, AST and ALP enzyme
activities, gradual significant reduction was observed in
serum of mice treated with deltamethrin and endosulfan
for two week. The reduction observed in AST and ALT
attributed to the hepatocellular damage resulting from
chemical-toxicity, where the transaminases levels
showed an intimate relationship to cell necrosis and /or
increased cell membrane permeability which led to the
discharge of enzyme to blood stream. The decrease in
transaminase levels providing additional support for the
side effect of the deltamethrin and endosulfan on
mitochondria of the hepatic cells as it is the subcellular
localization of transaminases (El Shazly et al., 2001).
961
Ismail et al., 2013
Fructose-1,6-diphos-phatase
(umole/mg protein/min.)
% change % Change
12.61.22
36.32% 15.4 1.11 22.22%
58.24% 21.3 1.43 69.1%
This was attributed to the irritation of liver cells by
toxins or due to increase loss of intracellular enzyme by
diffusion through cell membrane. In the present study,
acid phosphatase show significant elevation in serum
of treated mice. Higher levels of acid phosphatase in
tissue was observed by El-Aasar et al., (1989) and
Abdel-Rahman et al., (1993), which was attributed to the
of irritation of liver cells by toxins or metabolic products of
growing schistosomula of adult worms and eggs or due
to increase loss of intracellular enzyme by diffusion
through cell membrane which appear to act as a stimulus
to the synthesis of more enzyme.
Regarding the sources of energy for mice,
deltamethrin and endosulfan significantly decreased the
glycogen content in liver tissues of treated mice, while
the glucose level increased in the serum of treated mice.
This may be attributed to the activity of the pesticides
that impedes oxygen consumption of mice, thus inducing
anaerobic respiration. Under hypoxic conditions, animals
derive their energy from anaerobic breakdown of
glucose, which is available to the cells by increased
glycogenolysis (Vincent et al., 1995; Sambasiva, 1999).
Nakano and Tomlinson (1967) have suggested that
catecholamine levels rise under stressful environmental
conditions, enabling the increased utilization of glycogen
for energy production. To restore its energy
requirements, the mouse has to increase the rate of
glycolysis thus bringing about a reduction of the
glycogen content and increase glucose level in the blood
(Baskaran and Palanichamy, 1990; Vasanthi and
Baskaran, 1990).
Journal of Research in Biology (2013) 3(4): 954-966
Ismail et al., 2013
The data obtained in the present study showed
that lipid peroxide was elevated in the liver of mice
treated with deltamethrin and endosulfan.
peroxidation is known to require the participation of
highly reactive oxygen and other reactive metabolites in
the chain of biochemical reaction (Botros et al., 2007).
At the same time, liver GSH was drastically depleted in
the liver. Such depletion is critical, as shown by the
increased cytotoxicity of H2O2 in endothelial cells, as a
result of inhibition of glutathione reductase, which keeps
glutathione in its reduced state (El-Rigal et al., 2006).
In a good agreement with the present
Mittelstaedt et al., (2004) suggested that nuclei and
mitochondria act as major targets of MG toxic action,
probably by increasing the generation of free radicals,
lipid peroxidation and DNA adducts formation.
Total protein content showed significant
reduction in liver tissue of mice treated with deltamethrin
and endosulfan for 2 weeks. This could be attributed to
cellular damage caused by toxin (Parasad et al., 1991).
The significant decrease in total protein content is mainly
due to increase in messenger RNA degradation which is
the possible cause for the hypoalbuminemia (Metwally
et al., 1990).The depletion of the protein fraction in the
liver tissue of mice in this experiment may have been
due to interference of active substance of pesticides in
protein metabolism by inhibiting protein synthesis (Volpi
et al., 1997).
The current investigation revealed significant
enhancement of PK, PFK and GPI the rate limiting
glycolytic enzymes, in liver tissue of treated mice
with two pesticides compared to control group
accompanied by a marked increase in
gluconeogensis; F,1-6-diphosphtase. The increase in
gluconeogenic enzymes may be also responsible for the
production of glucose during treatment (Klover and
Mooney, 2004). Stimulation of PK in pesticide toxicity
ascertained the enhancement of glycolytic
previously reported by Ahmed and Gad (1995).
Journal of Research in Biology (2013) 3(4): 954-966
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 Journal of Research in Biology An International Scientific Research Journal

Original Research

Prevalence and the effect of plant extracts on community associated

methicillin resistant Staphylococcus aureus in Owerri, Imo State, Nigeria
Journal of Research in Biology

Authors: ABSTRACT:
Amadi ES1, Oguoma OI1, The prevalence of Methicillin resistant Staphylococcus aureus (MRSA) among
Ibekwe VI1, Abanobi SE2, apparently healthy inhabitants of Eziobodo Community and Students of Federal
Chikwendu CI1 and University of Technology Owerri (FUTO), Imo State, Nigeria was studied. The work
Egbadon OE1. further ascertained the antibacterial activities of medicinal plants including
Azadirachta indica, Pterocarpus mildbraedii, Garcinia kola, Phyllanthus amarus and
Vernonia amygdalina against the MRSA isolates. A total of two hundred nasal swab
specimens were randomly collected from the participants. The Kirby-Bauer technique
Institution: was used to determine the susceptibility pattern of the isolates to Vancomycin (5g),
1. Department of Ciprofloxacin (5g), Ceftriaxone (30g), Oxacillin (5g), Methicillin (10g) and
Microbiology, School of Erythromycin (15g). The antibacterial properties of the ethanolic plant extracts were
Science, Federal University determined using the agar well diffusion technique. A total of 181 (90.5%) and 141
of Technology, P.M.B. 1526, (70.5%) of the nasal swab samples, yielded Staphylococcus species and
Owerri Imo State, Nigeria. Staphylococcus aureus respectively. The antibiotic sensitivity screening revealed that
38 (27%) of the S. aureus isolates were methicillin resistant. The MRSA isolates also
2. Department of exhibited the highest resistance to vancomycin and the least to ceftriaxone.
Biochemistry, School of Furthermore, the result showed that crude ethanolic extracts of all tested plant
Science, Federal University extracts except Pterocarpus mildbraedii exhibited antibacterial activities against the
of Technology, P.M.B. 1526, MRSA isolates. Phytochemical components such as Alkaloids, Tannins, Glycosides,
Owerri Imo State, Nigeria. Saponins, Flavonoids, Terpenoids, Phlobatannins, Steroids and Anthraquinones were
detected in the plant materials in varying proportions. This study unveils a relatively
high occurrence of MRSA among the study population which could be a risk factor for
infection with MRSA. These plant extracts could also serve as potential sources of
therapy for the treatment of MRSA infections.

Corresponding author: Keywords:

Chikwendu CI. Antibiotic resistance, Staphylococcus aureus, Methicillin, Plant extracts,
Isolates.

Email: Article Citation:

Amadi ES, Oguoma OI, Ibekwe VI, Abanobi SE, Chikwendu CI and Egbadon OE.
Prevalence and the effect of plant extracts on community associated methicillin
resistant Staphylococcus aureus in Owerri, Imo State, Nigeria.
Journal of Research in Biology (2013) 3(4): 967-976

Dates:
Web Address: Received: 20 Mar 2013 Accepted: 09 May 2013 Published: 05 June 2013
http://jresearchbiology.com/
documents/RA0342.pdf.

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.

967-976 | JRB | 2013 | Vol 3 | No 4

Journal of Research in Biology
An International Scientific
Research Journal www.jresearchbiology.com

INTRODUCTION
Staphylococcus aureus is a coagulase positive,
gram positive cocci, which apart from being a normal
flora of the anterior nares, skin and large intestine, is also
capable of causing a wide range of diseases varying from
minor skin infections to life threatening septicemia,
pneumonia, endocarditis, deep-seated abscess among
others (Willey et al., 2008; Lowy 2003; Kuehnert et al.,
2006; Tenover and Gaynes, 2000; Holmes et al., 2005;
Nester et al., 2007). Penicillin and later methicillin were
very efficacious in the management of Staphylococcus
infections in the early 1960s. However, over the years,
most strains have acquired resistance to these drugs due
to acquisition of gene encoding the enzyme penicillinase.
In recent times, strains of S. aureus have emerged that
not only produce penicillinase, but also have Penicillin
binding proteins (PBPs) with low affinity for all -lactam
drugs. These strains referred to as methicillin resistant
Staphylococcus aureus (MRSA) are resistant to
m eth i cil l in an d oth er -l a cta m drugs
(Nester et al., 2007; Willey et al., 2008). Nearly all
MRSA have additional genetic material known as mec A
gene not found in methicillin sensitive strains, which
encodes PBP 2a, a cell wall transpeptidase, having
reduced affinity for -lactam antibiotics. The mec A
gene is found as a part of a mobile genetic element found
in MRSA strains known as Staphylococcal cassette
chromosome mec (SCC mec) (Jeshina and Surekha,
2009; Pinho et al., 2001).
In addition to lactam drugs, MRSA isolates
have become resistant to a number of antimicrobial
agents such as, fluoroquinolones, aminoglycosides and
macrolides (Shittu et al., 2009). MRSA could be
categorized as either hospital acquired (HA-MRSA) or
community acquired (CA-MRSA), depending on the
source of acquisition. While the former occur in
individuals who are/have recently been in a hospital or
other healthcare facility, the latter are acquired by
persons not recently hospitalized. According to David
968
Amadi et al., 2013
and Daum (2010), all infections occurring among
outpatient or among inpatients with an MRSA obtained
earlier than 48 hours after hospitalization could be
regarded as CA-MRSA. In addition, livestock associated
MRSA (LA-MRSA) have been reported to pose a
challenge particularly in countries with low level of
MRSA (Stefani et al., 2012). Morris et al., (2012)
reported the potential for pet animals to harbour MRSA
when residing with human MRSA patients.
The fact that MRSAs are becoming more
prevalent worldwide and also resistant to a wide range of
antibiotic groups, underlines the need for alternate
strategies to stem the immense public health challenge
posed by these organisms. Natural products from local
medicinal plants are increasingly being used in the
treatment of many hard to treat diseases and the search
for more potential compounds from plants has continued
(Lai et al., 2010; Newman and Cragg, 2007). According
to WHO, 65-80% of the world population rely on
traditional medication for their ailments (Gurinder and
Daljit, 2009). A number of works has highlighted the
efficacy of local indigenous plants against a wide range
of pathogens (Ugbogu et al., 2010; Lai et al., 2010;
Aliyu et al., 2008; Aliyu et al., 2011; Ajibade et al.,
2010). The present study was aimed at ascertaining the
occurrence of MRSA among apparently healthy
Eziobodo community inhabitants and FUTO students as
well as their susceptibility to different antibiotic groups.
It also determined the antibacterial effects of some local
plant extracts on the MRSA isolates.
MATERIALS AND METHODS
Collection of nasal specimens
Two hundred (200) nasal specimens were
collected, 100 each from the anterior nares of apparently
healthy individuals of Eziobodo community (one of the
communities hosting FUTO) and students of Federal
University of Technology (FUTO), all in Owerri West
LGA, Imo State, Nigeria. They were aseptically
Journal of Research in Biology (2013) 3(4): 967-976
Amadi et al., 2013
collected using sterile swab sticks between August and
November 2010.
Cultivation and isolation of Staphylococcus aureus
The respective nasal specimens were cultivated
within one hour of collection in Mannitol salt agar and
nutrient agar using standard techniques to obtain discreet
colonies. The plates were incubated at 37oC for 24 hours.
The axenic cultures of the isolates were subsequently
identified using colony morphology, microscopy and
biochemical tests including catalase and coagulase tests
(Cheesbrough, 2002).
Antibiotic susceptibility test
The antibiotic susceptibility screening of the
S. aureus isolates was conducted using the Kirby-Bauer
disc diffusion method (Cheesbrough, 2002). Standard
inoculum, equivalent of 0.5 McFarland standards of the
isolates was evenly spread on Mueller Hinton agar
plates. Antibiotic discs including Vancomycin (5g),
Ciprofloxacin (5g), Ceftriaxone (30g), Oxacillin
(5g), Methicillin (10g) and Erythromycin (15g)
(Oxoid) were aseptically placed on the plates. The plates
were then incubated at 37oC for 24 hours and the
inhibition zones recorded in millimeters using a meter
rule.
Subsequently, all the isolates identified as
Methicillin resistant Staphylococcus aureus (MRSA)
were subjected to antibiotic screening test using the same
disc diffusion technique as above. The following
antibiotics were used; Vancomycin (5g), Ciprofloxacin
(5g), Ceftriaxone (30g) and Erythromycin (15g)
(Oxoid).
Collection of plant materials
The leaf and bark of Pterocarpus mildbraedii,
Azadirachta indica, leaves of Vernonia amygdalina, and
whole plant of Phyllanthus amarus were obtained from
the premises of FUTO. The seeds of Garcinia kola
however, were purchased from Ekeukwu Owerri market,
Imo State. The plant materials were subsequently
authenticated by a taxonomist.
Journal of Research in Biology (2013) 3(4): 967-976
Figure 1: Antimicrobial resistance rates (%) of
MRSA isolates to different antibiotics
Preparation of plant extracts
The leaves, barks and seeds of the plants were
washed and dried at room temperature and later
pulverized. 20gm of each plant powder was separately
mixed with 250ml of ethanol and the extraction was done
using the soxhlet extraction procedure.
Phytochemical screening
The phytochemical screening of each plant
extract was carried out to determine the presence or
absence of Alkaloids, Tannin, Saponins, Glycosides,
Anthraquinone, Steroids, Flavonoids, Terpenoids, and
phlobatannins (Harbone, 1973; Sofowora, 1993).
Susceptibility screening of MRSA isolate to plant
extracts
The antibacterial effects of each plant extract on
MRSA were determined using the agar well diffusion
technique (Perez et al., 1990). Standard inoculum,
equivalent of 0.5 McFarland standards of the isolates was
evenly spread on Mueller Hinton agar plates. Sterile cork
borer was used to make wells on the agar. The
reconstituted extracts (25mg/ml, 50mg/ml, 100mg/ml
and 200mg/ml) were respectively introduced into wells
and labeled accordingly. Following the incubation of the
plates at 37oC for 24 hours, the inhibition zone diameters
were recorded using meter rule.
969

Table 1: Prevalence (%) of Staphylococcus aureus and MRSA
isolates from Eziobodo and FUTO inhabitants
Target population No of samples Staphylococcus sp.
Eziobodo 100 95(95)
FUTO students 100 86(86)
Total 200 181(90.5)
RESULTS AND DISCUSSION
Out of the total of 200 specimens collected, 181
isolates were identified as Staphylococcus species, while
141 isolates were identified as S. aureus, representing a
prevalence rate of 90.5% and 70.5% respectively
(Table 1). Also, the antibiotic resistance screening of the
isolates showed that 38 (27%) of the S. aureus isolates
were MRSA. The MRSA isolates exhibited their highest
sensitivity to Ceftriaxone and the least to Vancomycin
antibiotic (Table 2 and Figure 1).
The Phytochemical screening of the plant
extracts revealed the presence of Phytochemical
components such as alkaloids, saponins, flavonoids and
others in varying quantities (Table 3).
The antibacterial screening of the ethanolic
extracts of the medicinal plants used in this study
indicated that all except Pterocarpus mildbraedii,
exhibited inhibitory activity against MRSA isolate.
(Table 4).
The result of this study showed that 90.5% and
70.5% of the isolates from the anterior nares of Eziobodo
inhabitants and FUTO students were respectively
Staphylococcus species and Staphylococcus aureus. This
is consistent with the findings of Ugbogu et al., (2010)
and Chigbu and Ezeronye (2003), but higher than the
33.3% prevalence reported from Amassoma community
in Niger Delta, Nigeria (Onanuga and Temedie,
2011).This high occurrence in our present work is not
unexpected, since S. aureus is a normal microflora of the
human body, particularly the upper respiratory tract
(Willey et al., 2008; Cheesbrough 2002).
970
Amadi et al., 2013
Staphylococcus aureus MRSA
66(66) 20(30.3)
75(75) 18(24)
141(70.5) 38(26.9)
The Methicillin resistant Staphylococcus aureus
(MRSA) prevalence rate of 27% among apparently
healthy individuals as reported in the present work is
considerably low compared to a report by Ugbogu et al.,
(2010) who isolated 83.5% of MRSA from healthy
individuals in Abia State, South East Nigeria and
Onanuga et al., (2005) that recorded 69% isolation from
healthy women in Zaria, Nigeria. Similarly, our current
finding is also lower than the report of Olowe et al.,
(2007) and Onanuga and Temedie (2011), in which
47.8% and 47.6% MRSA were isolated in South West
and Niger Delta regions of Nigeria respectively. The
prevalence rate of 47.15% and 43% has also been
reported from Ibadan and Jos, Nigeria (Ghebremedhin
et al., 2009; Ekeh, 2003). However, the current result is
consistent with the report of Nwankwo et al., (2010) in
which 28.6% was recorded. The difference in the
prevalence of MRSA obtained in the present study and
those of previous works could be attributable to strain
variation in different geographical regions and locations
(Ikeagwu et al., 2008).
It is important to note that the recovery of MRSA
from apparently healthy community inhabitants in the
present study is very significant particularly at this time
when infiltrations of Community acquired MRSA
(CA-MRSA) to healthcare facilities has been reported in
some parts of the world (Stefani et al., 2012). According
to Creech et al., (2005), Farley et al., (2008), and
Hidron et al., (2005) enormous reservoirs of MRSA now
exist outside health care settings and this implies that the
current methods of MRSA control in health facilities are
not likely to succeed. In this regard, preventive measures
Journal of Research in Biology (2013) 3(4): 967-976
Amadi et al., 2013

Table 2: Frequency (%) of antibiotic resistance S. aureus isolates from nasal

samples of Eziobodo and FUTO inhabitants
No (%) of resistant isolates
Eziobodo FUTO
Total no of
No of No of resistant No of No of resistant Total no of
Antibiotics resistant
isolates isolates isolates isolates isolates
isolates
Oxacillin 66 21(31.8) 75 20(26.7) 141 41(29.1)
Methicillin 66 18(27.3) 75 20(26.7) 141 38(27)
Ciprofloxacin 66 16(24.2) 75 10(13.3) 141 36(25.5)
Vancomycin 66 15(22.7) 75 7(9.3) 141 22(15.6)
Erythromycin 66 10(15.1) 75 9(12) 141 19(13.5)
Ceftriaxone 66 3(4.5) 75 0(0) 141 3(2.1)

Table 3: Phytochemical components of plant extracts

Phytochemical components
Plant
Alkaloids Tannins Glycosides Saponinss Flavonoids Phlobatannins Steroids Anthraquinones Terpenoids
extracts
AIL + + - + + - + + -
AIB - + + + + + - + +
PML + + + + + - - - -
PMB - + + - - + - + +
PA - + - - - - + + -
VA + + + + + + -
GA - + + + - + - + +
Key: AIL Azadirachta indica Leaf, AIB Azadirachta indica Bark, PA Phyllanthus amarus, PML Pterocarpus mildbraedii
Leaf, PMB Pterocarpus mildbraedii Bark, VA Vernonia amygdalina, GK Garcinia kola.

to stop the possible transmission in the communities is a
viable approach ( Charlebois et al., 2004; Cooper et al.,
2004; David et al., 2008; Liu et al., 2008).
The antibiotic susceptibility test revealed that all
the S. aureus isolates exhibited the least resistance to
ceftriaxone antibiotic in the present study. This finding is
consistent with the report of Masood and Aslam (2010)
in which 96.1% susceptibility of S. aureus isolates to
ceftriaxone was highlighted. Ceftriaxone was apparently
recommended by these workers as a drug of choice for
infections caused by S. aureus, Escherichia coli,
Pseudomonas aeruginosa, Klebsiella pnuemoniae and
Salmonella typhi. On the other hand, the S. aureus
isolates were more resistant to Ciprofloxacin and
Erythromycin (Tables 2 and 3). The resistant rates are in
line with the reports of Shanhraz et al., (2012), and
Onanuga and Temedie (2011), but quite low compared to
the over 70% resistance recorded by Ojulong et al.,
(2009) in Kampala, Uganda. Azeez-Akande et al.,
(2008), however reported a susceptibility rate of 93.9%
of MRSA isolates to ciprofloxacin.
Furthermore, vancomycin has been described as
a reliable alternative for the treatment of MRSA
infections. Elhamzaoui et al., (2009) and Nwankwo and
Nasiru (2011) reported 100% sensitivity of S. aureus
isolates from a University hospital in Rabat Morocco and
a tertiary health institution in Kano, Nigeria, to
Vancomycin respectively. Nevertheless, this antibiotic,
vancomycin, which was initially a drug of choice in the
treatment of MRSA infections, is witnessing resistance
in recent times (Von-Eiff et al., 2001). In the present
work, over 50% of the MRSA isolates were resistant to
Vancomycin. This is worrisome because Vancomycin
has been described by various workers as very effective

Journal of Research in Biology (2013) 3(4): 967-976 971

 Amadi et al., 2013

Table 4: Inhibitory activities of plant extracts concomitant effect of the active compounds contained by
against MRSA isolates these plants on MRSA. However, none of the extracts
Mean zone of inhibition (mm)/ were active against the MRSA isolates at the lowest
Concentration of plant extracts (mg/ml)
concentration of 25mg/ml (Table 4).
Plant
25 50 100 200 The inhibitory effect of Garcinia kola extract on
extract
AIL - - - 9 MRSA as observed in the present study is in agreement
AIB - 7 8 12
with the work of Ugbogu et al., (2010) and
PML - - - -
PMB - - - - Adeleke et al., (2006), in which Garcinia kola extracts
PA - - - 8 exhibited antibacterial activities against MRSA isolates
VA - - 9 10 in Nigeria. Also, Taiwo et al., (1999) reported that
GK - - - 11
Garcinia kola exhibited strong activity against MRSA.
Key: AIL Azadirachta indica Leaf, AIB Azadirachta indica
Bark, PA Phyllanthus amarus, PML Pterocarpus Similarly, the inhibitory properties of Azadirachta indica
mildbraedii Leaf, PMB Pterocarpus mildbraedii Bark,
VA Vernonia amygdalina, GK Garcinia kola. and Vernonia amygdalina against MRSA as recorded in
the present work is consistent with the reports of
against MRSA and in fact a drug of choice in the Skariyachan et al., (2011), Aliyu et al., (2011)
treatment of multidrug resistant S. aureus infections and Aliyu et al., (2008). Furthermore, that
(Ojulong et al., 2009; Elhamzaoui et al., 2009). The Phyllanthus amarus extract had antibacterial activity
vancomycin resistance rate as recorded in the current against MRSA is in line with the findings of
study is however contrary to the report of Onanuga and Aliyu et al., (2008). Ajibade et al., (2010) also
Temedie (2011) in Niger Delta Nigeria and highlighted the antimicrobial activity of Phyllanthus
Shanhraz et al., (2012) in which over 70% susceptibility species against MRSA. Undoubtedly, the findings of this
was recorded. The present finding thus suggests that study support the local use of these plant materials in the
vancomycin may be inefficient in the treatment of treatment of most hard to treat infections.
infections caused by MRSA in the near future among our In conclusion, the recovery of CA-MRSA from
target population. The present study therefore the external nare of apparently healthy individuals in this
recommends ceftriaxone as a drug of choice for the study underscores the significance of the nasal region as
treatment of MRSA infections in our study area. a reservoir of S. aureus, and by implication MRSA. In
The increasing resistance of MRSA to -lactam fact, MRSA colonization of the nares is believed to be a
and other broad spectrum antibiotics has stimulated risk factor for a clinically apparent infection with MRSA
recent investigations on plant parts for naturally (Croft et al., 2009; Huang et al., 2006; Lu et al., 2007;
occurring active compounds as alternatives to treatment Muder et al., 1991). It is therefore very imperative that
of MRSA caused infections. The phytochemical strategies should be designed to halt the further spread of
screening of the plant extracts used in this study revealed MRSA in communities and most especially to
the presence of alkaloids, Tannins, saponins, flavonoids, immunodeficient individuals. According to
terpenoids, anthraquinones, glycosides and steroids Stefani et al., (2012), CA-MRSA clones spreading in the
(Table 4). The antibacterial screening of the plant community could also infiltrate healthcare facility in
extracts showed that all the plant materials used except many parts of the world. This certainly would exacerbate
Pterocarpus mildbraedii exhibited inhibitory activity the challenges already posed by MRSA. Interestingly
against MRSA. This effect could be attributed to the however, the therapeutic activities of the plant materials
972 Journal of Research in Biology (2013) 3(4): 967-976
Amadi et al., 2013

used in this study could hold a great promise as a Chigbu CO and Ezeronye OU. 2003. Antibiotic
potential precursor in the development of therapies for resistant Staphylococcus aureus in Abia State of Nigeria.
the management of MRSA infections, if properly Afr. J. Biotechnol., 2(10): 374-378.
harnessed.
Cooper BS, Medley GF, Stone SP, Kibber CC,
Cookson BD, Roberts JA, Duckworth G, Lai R and
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 Journal of Research in Biology An International Scientific Research Journal

Original Research

Hepatic enzyme markers and proteins in serum and some selected tissues
in Clarias gariepinus from swamp around Kokori-Erhoike oil field, Nigeria
Journal of Research in Biology

Authors: ABSTRACT:
Osioma E1*, Akanji MA1 This study determines changes in some biochemical parameters in serum and
and Arise RO1. tissues of Clarias gariepinus obtained from fish natural habitat in the oil exploration
environs of Kokori-Erhoike in Delta State, Nigeria. Sampling sites include Ethiope River
(Eku axis, reference Site A); Erhoike swamp (Site B) and Erhoike fish pond
(Site C). However, Sites B and C are located in the oil exploration region of Erhoike.
Clarias gariepinus (n=8) were collected from each site and used for the study. Levels of
total proteins, albumin, haemoglobin as well as the activities of alanine
aminotransferase (ALT), aspartate aminotransferase (AST) and alkaline phosphatase
were determined in serum, gill, liver, brain and muscle tissues. Results showed that
total protein concentrations were comparable (p>0.05) in serum. Albumin
Institution: concentrations of fish from Site B and C were lower (p<0.05) as compared with that of
1. Department of site A in gill, muscle and brain tissues. Haemoglobin concentration was significantly
Biochemistry, Faculty of lower (p<0.05) in fish from Site A as compared with that of Sites B and C. Results also
Science, University of Ilorin, indicated that total protein and albumin concentrations were significantly (p<0.05)
Nigeria. higher in gill, muscle and brain tissues of Clarias gariepinus from Site A as compared
with fish tissues from Sites B and C. Hepatic enzymes (ALT and AST) and ALP activities
were elevated (p<0.05) in serum, gill, brain and liver of fish from Sites B and C as
compared with that of Site A. The observed biochemical changes in fish from Sites B
and C could have resulted from contaminants arising from the oil exploration activities
in Site B and the presence of organic/inorganic contaminants in Site C due to the
presence of fish feeds. These biochemical alterations show that the fish were under
stress in their natural habitat. These biomarkers could be employed in the
environmental monitoring of crude oil pollution as well as early warning signs of the
adverse effects of environmental pollution.

Corresponding author: Keywords:

Osioma E. Fish, Kokori-Erhoike, Clarias gariepinus, Albumin, Alkaline phosphatase,
Haemoglobin.

Email: Article Citation:

Osioma E, Akanji MA and Arise RO.
Hepatic enzyme markers and proteins in serum and some selected tissues in
Clarias gariepinus from swamp around Kokori-Erhoike oil field, Nigeria.
Journal of Research in Biology (2013) 3(4): 984-992

Web Address: Dates:

http://jresearchbiology.com/
Received: 23 Mar 2013 Accepted: 23 May 2013 Published: 13 June 2013
documents/RA0343.pdf.

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.

984-992 | JRB | 2013 | Vol 3 | No 4

Journal of Research in Biology
An International Scientific
Research Journal www.jresearchbiology.com

INTRODUCTION
Over the years, the extent of oil exploration
activities and its related environmental effects has been
on the increase, (Tolulope, 2004). Some quantities of
petroleum and its products may be released into the
environment during oil exploration activities due to
operational, accidental, transportation or other means.
Apart from contaminating the flesh of commercially
valuable fish, crude oil compromise fish hatcheries in
coastal waters (Leighton, 1991) and its products are the
most relevant to aquatic ecotoxicology (Pacheco and
Santos, 2001). In Nigeria, crude oil was discovered at
Oloibiri in 1959 (Akpofure et al., 2000) and exploration
activities has been carried out in Kokori- Erhoike
environment for over 30 years (Emoyan, 2009).
The cause-effect relationship and result of
xenobiotic pollution in an ecosystem can be assessed
through the analysis of biochemical alterations on
organisms inhabiting that environment. These
biochemical alterations may be sensitive and specific as
early indicators of aquatic pollution (Norris et al., 2000;
Strinac and Braunbeck, 2000)
Protein plays a vital role in the physiology of
living organisms and its metabolism according to Adams
et al., (1990) provides information on the general energy
mobilization of an animal and show relationship with
effects of contaminants in these organisms. The
concentration of plasma albumin is a useful index of the
state of protein repletion and it makes the major
contribution to plasma sulphydryl groups which can
function as a chain breaking antioxidant (Halliwell,
1988). Haemoglobin contained in the red blood cells
which serve as the oxygen carrier in blood has been
employed in assessing the health of fish and monitoring
stress response of several environmental contaminants
including petroleum hydrocarbons (Soivio and Oikari.,
1976; Gabriel et al., 2007).
Alanine aminotransferase (ALT) and aspartate
aminotransferase (AST) catalyze the transfer of -amino
985
Osioma et al., 2013
group from -amino acid to -keto acid. AST and ALT
are biological responses of severe hepatic injury and
their bioassay can serve as a diagnostic tool for
estimating necrosis of the liver cells. (Cappo et al.,
2002). The determination of ALT and AST activities has
been applied in fish research to indicate bacteria, viral
and parasitic infection, intoxications and water pollution
(Bucher and Hofer, 1990). Alkaline phosphatase
comprises group of enzymes which is responsible for
hydrolyzing phosphoric ester bonds present in organic
compounds at an alkaline pH (Akcakaya et al., 2007).
The enzyme (ALP) has been reported to be a marker
enzyme for the plasma membrane and endoplasmic
recticulum (Akanji et al., 1993).
Fish species are excellent subjects for the study
of various effects of contaminants (El-Shehami et al.,
2007) and African catfish (Clarias gariepinus) has been
used in fundamental research and toxicological studies
(Nguyen and Janssen, 2002)
A significant body of research has investigated
the effects of crude oil (or its derivatives) and refinery
effluents on fish health. The investigators include:
Yarbrough et al., 1976; Kuehn et al., 1995; Sunmonu
and Oloyede, 2006; Wegwu and Omeodu, 2010;
Mahmoud et al., 2011 and Nwaogu et al., 2011.
However, there is dearth of information on the effect of
crude oil exploration activities on African catfish
(Clarias gariepinus) obtained from swamps (fish natural
habitat) around Kokori- Erhoike oil field located in Delta
State, Nigeria. This information deficiency prompted this
study. Therefore, the aim of this research is to determine
changes of some biochemical parameters in serum and
tissues (gill, liver, brain and muscle) of
Clarias gariepinus obtained from swamps around Kokori
- Erhoike petroleum flow station in Delta State, Nigeria.
MATERIALS AND METHODS
Sampling sites:
Journal of Research in Biology (2013) 3(4): 984-992
Osioma et al., 2013

This study was carried out in Ethiope East Local
Government Area of Delta State, Nigeria. Experimental
areas have been previously described by Aries et al.,
2013 and are represented in Figure 1 below.
Site A (reference site) is the Eku axis of the
Ethiope River, Delta State. There is no presence of oil
facilities/operations or any industry located along the
Ethiope River from its source, Umuaja, about 22km to
the Eku axis. The upper axis of the Ethiope River has
been reported to be relatively unpolluted (Ikomi et al.,
2005; Agbaire and Obi, 2009; Aries et al., 2013). This
qualifies the Eku axis of Ethiope River as a reference site
for this study. Site B is the swampy environment of
Kokori-Erhoike petroleum flow station where oil
exploration activities have been on for more than 35
years. This area has a number of oil wells and flow
stations. The aquatic ecosystem in the area is constituted
by non-tidal freshwater swampy forest characteristics of
those found within the freshwater survey zone of the
Niger Delta. Site C is a natural fish pond located within
Kokori-Erhoike environment. The main uses of water in
the catchments include domestic, recreational (e.g.
swimming) and fishing. Their major occupation includes
farming (cassava, yam, Okro etc.), fishing and petty
trading on food stuff.

Fig 1. Map of Ethiope East showing the location sampling sites

Source: Ministry of lands, Surveys and urban Development, Asaba. (2008)

Journal of Research in Biology (2013) 3(4): 984-992 986

 Osioma et al., 2013

Fish Duncans Multiple Range Test (DMRT) at 5%

Eight African catfish (Clarias gariepinus) were probability level. All statistical analysis was performed
collected from each site in October, 2011. At site A (Eku using SPSS version 16.
River) and site B (Erhoike swamp), fish were caught
with the help of professional local fishermen and at site RESULTS
C (Erhoike Fish pond) fish net was used to catch the fish. Results (Table 1) show that gill, muscle and
Preparation of Serum brain total protein concentrations were significantly
About 1.5ml blood was taken by caudal arterial higher (p<0.05) in fish from Site A as compared with
puncture from each fish into a sterilized plain tube. that of Site B and C. Total protein concentration in serum
Blood was allowed to clot for about 5min dislodged and and liver of Clarias gariepinus from all sites were
centrifuged at x15,000g for 15min to obtain the serum, comparable at p>0.05. The data also indicated significant
which was stored frozen at 20C until analyzed. reduction (p<0.05) in gill and muscle albumin
Preparation of Tissue Homogenate concentration of Clarias gariepinus from Sites B and C
The fish were dissected and the gill, liver, brain as compared with that of Site A. Levels of serum and
and muscle tissues were quickly removed. The tissues liver albumin were comparable in fish from all sites.
(gill, liver, brain and muscles) were washed in cold Results in Table 1 also revealed that fish from Sites B
saline (0.9% NaCl) solution several times and then 1g of and C has elevated (p<0.05) haemoglobin concentration
wet tissue was homogenized in 9 ml of the physiological compared with that of Site A.
solution (normal saline). The resulting homogenate was
centrifuged at 5000g for 20min. The supernatant was
decanted and used for further biochemical analysis. Table 1: Levels of total protein, albumin and
haemoglobin concentrations in serum and tissues (gill,
Biochemical Investigations
liver, muscle and brain) of Clarias gariepinus from
The concentrations of total protein and albumin swamps around Kokori-Erhoike Petroleum Flow
Station in Delta State, Nigeria
were determined in serum and tissues (gill, liver, muscle
and brain) employing the methods of Doumas et al., SAMPLING SITES
A B C
1981 and Doumas et al., 1971 respectively. Total protein concentration (g/dl)
Haemoglobin level was estimated by the method of Tietz Serum (n=8) 9.310.53a 9.300.31a 9.290.33a
Gill (n=8) 2.820.05a 2.460.24b 2.430.03b
(1976), while the method of Roy (1970) was used to Liver (n=8) 2.810.37a 2.910.15a 2.980.40a
determine the activities of alkaline phosphatase. Alanine Muscle (n=8) 5.280.32a 4.290.10b 4.080.40b
Brain (n=8) 4.170.17a 3.720.13b 3.960.12c
aminotransferase and aspartate aminotransferase were Albumin concentration (g/dl)
analyzed using the method of Reitman and Frankel Serum (n=8) 7.250.32a 7.010.01a 6.960.13a
Gill (n=8) 2.480.36a 2.030.16b 2.080.24b
(1957). All assays were carried out with the aid of Liver (n=8) 2.520.43 a
2.510.26 a
2.690.18a
a b
commercially available kits supplied by TECO Muscle (n=8) 4.090.17 2.090.42 2.550.12b
Brain (n=8) 3.780.22a 2.250.11b 3.680.10a
Diagnostics, Anahem, USA and Randox Laboratories,
Haemoglobin concentration (g/dl)
Ardmore, United Kingdom. Blood (n=8) 22.241.57a 27.031.65b 27.291.65b
Statistics
Values are given as Mean SD. Means not sharing a
Analysis of Variance (ANOVA) was used to
common superscript letter on a given row differ
analyse data obtained from the various biochemical significantly at p<0.05. A= Ethiope River (Eku axis);
B= Erhoike swamp; C= Erhoike fish pond.
investigations. Group means were compared by the
987 Journal of Research in Biology (2013) 3(4): 984-992
Osioma et al., 2013
Increased (p<0.05) activities of alanine
aminotransferase in serum and liver of
Clarias gariepinus from Sites B and C were observed as
compared with that of Site A (Table 2). Similar trend
was observed for the activity of liver aspartate
aminotransferase. The table also showed that the activity
of aspartate aminotransferase in serum of fish from all
sites (A, B and C) were comparable (p>0.05), although,
fish from Site A had relatively lower aspartate
aminotransferase activity. Results (Table 2) also showed
that the activity of alkaline phosphatase in (serum, liver,
gill and brain) of Clarias gariepinus from Sites (B and
C) were significantly (p<0.05) higher as compared with
that of Site A.
DISCUSSION
Biochemical markers of pollution are considered
indicators employed in fish toxicity tests and for field
monitoring of aquatic contamination. They established
contact of the sample with definite groups of chemical
compounds and clarify their metabolic fate. Biochemical
investigations allow cause-effect relationship to be
established at an early stage of pollution and these
sensitive and predictive diagnostic tools (Biomarkers) for
assessing animal exposure and toxic effects of chemical
contaminants are needed as aquatic environmental
contamination assessment indicators.
The total protein concentration in serum, gill,
liver, muscle and brain of Clarias gariepinus from Eku
River is higher than the values obtained from fish
samples in Erhoike swamp and Erhoike fish pond. The
differences were significant (p<0.05) in the gill, brain
and muscle tissues.
Proteins play a vital role in the physiology of
living organisms and provide information on the general
energy mobilization of an animal and show relationship
with the effect of contamination in these organisms
(Adams et al., 1990). The over 30 years of petroleum
exploration activities in Erhoike vicinity could have
Journal of Research in Biology (2013) 3(4): 984-992
Table 2: Changes in the activities of alanine
aminotransferase (ALT), aspartate aminotransferase
(AST) and alkaline phosphatase (ALP) in serum and
tissues (liver, gill and brain) of Clarias gariepinus
collected from swamps around Kokori-Erhoike
Petroleum Flow Station in Delta State, Nigeria
SAMPLING SITES
A (n=8) B (n=8) C (n=8)
Alanine aminotransferase (IU/L)
Serum 43.862.79a 48.280.34b 49.190.65 b
Liver 25.100.80a 45.901.63b 42.381.82c
Aspartate aminotransferase (IU/L)
Serum 53.758.84 a 54.342.88 a 54.253.15 a
Liver 52.310.79a 69.701.63b 65.941.82c
Alkaline phosphatase (IU/L)
Serum 32.863.14 a 46.223.05 b 45.763.49 b
Liver 43.011.48a 52.671.79b 53.832.91b
Gill 29.733.73a 36.791.39b 32.745.36a
Brain 52.942.36a 57.341.21b 60.372.25c
Values are expressed as Mean SD. Means not sharing a
common superscript letter on a given row differ
significantly at p<0.05. A= Ethiope River (Eku axis);
B= Erhoike swamp; C= Erhoike fish pond.
contaminated the aquatic environment and petroleum
hydrocarbon can act as a mediator in free radical
generation in fish (Achuba and Osakwe, 2003). During
stress conditions, fishes need more energy to detoxify the
toxicants and to overcome stress, thus carbohydrate
reserve is depleted to meet energy demand (Nelson and
Cox, 2005; Sudhanshu and Ajay, 2009). Since fish have
a very little amount of carbohydrate, the next alternative
source of energy is protein to meet the increased energy
demand occasioned by a pollutant.
The decrease in total protein level observed in
the gill, brain and muscle tissue could be to meet the
higher energy demands for metabolic purposes due to the
presence of petroleum hydrocarbon in Erhoike
environment and could also be related to impaired food
intake, increased energy cost of homeostasis, tissue
repair and detoxification mechanism during stress.
Albumin is the most soluble and most electrically
mobile of all the major serum protein components and it
is synthesized entirely by the hepatic parenchymal cells.
Lower albumin concentration was observed in the serum,
988

gill, liver, muscle and brain tissues of Clarias gariepinus
from Erhoike swamp and Erhoike fish pond compared
with corresponding albumin level of Clarias gariepinus
from Eku River (control site). The utilization of proteins
as an alternative source of energy by fish in stress
condition could have accounted for the reduced albumin
level in fish from Erhoike swamp and Erhoike fish pond.
Albumin has also been regarded as an antioxidant
molecule. It reacts with and neutralizes peroxyl radicals
(Stocker and Frei, 1991) and it is considered as a
sacrificial molecule that prevents damage when it acts as
an antioxidant because albumin is destroyed in the
process (Halliwell, 1988). Therefore, the observed
reduction of albumin concentration in African Catfish
from the oil exploration areas may be linked to its
participation as an antioxidant molecule to quench free
radical reactions in other to mitigate the impact of
oxidative stress or its utilization as a source of energy by
the fish in stress condition.
This study showed that Clarias gariepinus from
Erhoike swamp and Erhoike fish pond have higher levels
(p<0.05) of haemoglobin as compared with haemoglobin
concentration of Clarias gariepinus from the control site
(Eku River). Elevated levels of haemoglobin observed in
African catfish from Erhoike swamp and fish pond could
be as a result of stress induced by the presence of crude
oil and other contaminants (as in the case of the fish
pond) that leads to environmental hypoxia as a result of
chronic exposure to the contaminants and anaerobic
condition which lead to increase haemoglobin
concentration as a compensatory mechanism for
increased oxygen demand. This result corroborates with
the findings of Mdegela et al., (2010) who reported
significant elevation of haemoglobin concentration in
fish from Mzumbe sewage water. Zaki et al., (2010) also
reported significant increase in haemoglobin levels in
Tilapia zilli exposed to acute lethal concentration dose of
lead (Pb).
989
Osioma et al., 2013
Alanine aminotransferase and aspartate
aminotransferase (ALT and AST) are enzymes directly
associated with the conversion of amino acids to keto
acids. Apart from being considered to be important in
assessing the state of the liver and some other organs
Verma et al., (1981), transamination of the same
represents one of the main pathways for synthesis and
deamination of amino acid, thereby allowing interplay
between carbohydrate and protein metabolism during the
fluctuating energy demands of the organisms in various
adaptive situations. ALT and AST activities are direct
indicators of intense hepatic damage, thus their bioassay
can assist as a diagnostic tool for determining necrosis of
the liver cells (Whitehead et al., 1999; Cappo et al.,
2002). Ugwu et al. (2008) concluded that AST enzyme
activity in Heterobranchus bidorsalis adults could be
used as biomarker for monitoring crude oil pollution in
Nigeria.
C om p a r e d wi t h th e c on t r ol (i . e .
Clarias gariepinus from Ethiope River, Eku axis) the
activities of ALT and AST in serum and liver of
Clarias gariepinus from Erhioke swamp and Erhoike
fish pond were higher. Such increase of ALT and AST
may be partly due to hepatic damage resulting from
petroleum pollution (in case of Erhoike swamp) or
organic/inorganic contaminants (present in Erhoike fish
pond, Arise et al., 2013) induced oxidative insults on
the hepatocytes. In addition, increased protein catabolism
might be responsible for the elevation of these
transaminases. These results agree with the findings of
Ayalogu et al., (2001); Orisakwe et al., (2005).
In this study, serum, liver, gill and brain alkaline
ph osph a t a s e a ct i vi t y wer e m ea sur ed in
Clarias gariepinus from the three sampling sites. Marked
increase in ALP activity was recorded in the serum, liver,
gill and brain tissues of Clarias gariepinus from Erhoike
swamp and Erhoike fish pond as compared with ALP
activity of Clarias gariepinus from the control site (Eku
River). ALP together with ALT and AST provide an
Journal of Research in Biology (2013) 3(4): 984-992
Osioma et al., 2013
indication of the degree of inflammation as well as
possible causes of hepatocellular damage as well as
distortion of the plasma membrane and endoplasmic
recticulum. Results of the research agree with the
findings of Yarbrough et al., (1976) and Ayalogu et al.,
(2001).
CONCLUSION
The study demonstrates that the level of
contamination is enough to cause changes in the protein,
albumin and haemoglobin level with attendant effect on
the integrity of hepatocytes as evidence in the elevated
activities as seen in liver marker enzymes and alkaline
phosphatase activity of Clarias gariepinus from Erhoike
swamp and Erhoike fish pond. The above biochemical
changes showed that the fish were under stress in their
natural habitat (Erhoike swamp) thus; these markers
could be employed in the environmental monitoring of
crude oil pollutant and their associated metabolic
changes as early warning signs of adverse effects of
environmental pollution.
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Journal of Research in Biology (2013) 3(4): 984-992 992

 Journal of Research in Biology An International Scientific Research Journal

Original Research

Influence of the growing area on oil palm (Elaeis guineensis) inflorescences

insects population
Journal of Research in Biology

Authors: ABSTRACT:
Koua Kouakou Herv1,
Akpesse Apka Alexandre
Moise2, Tuo Yalamoussa3, Oil palm tree grows naturally on low ground and on plain. Seed production
and Hala Nklo4. varies from one area to another on the same oil palm plantation. Pollination of oil
palm is essentially entomophilous; it appeared useful to assess the influence of the
Institution:
growing area on the fluctuation of pollinating insects population. Samplings were
1. Flix Houphouet-Boigny
University of Cocody performed each month on male and female inflorescences during two years on plots
(Abidjan, Cte dIvoire). in lowland and plain. The insects showed no qualitative change from one area to
22 BP: 1611 Abidjan 22. another. Sixteen species of insects were observed on the male inflorescences against
10 species on female inflorescences. The inflorescences showed variation in the
2. Flix Houphouet-Boigny number of insects based on the growing area and the stage of flowering.
University of Cocody
(Abidjan, Cte dIvoire).
BP: 582 Abidjan 22.

3. Flix Houphouet-Boigny
University of Cocody
(Abidjan, Cte dIvoire). Keywords:
BP: 582 Abidjan 22. culture area; pollinating insects; Lam; Cte dIvoire.
4. National Center of
Agronomic Research
(CNRA, Cte dIvoire)
BP: 1740 Abidjan 01.

Corresponding author: Article Citation:

Koua Kouakou Herv.
Koua Kouakou Herv, Akpesse Apka Alexandre Moise, TUO Yalamoussa,
and Hala Nklo.
Influence of the growing area on oil palm (Elaeis guineensis) inflorescences
insects population.
Journal of Research in Biology (2013) 3(4): 940-946

Web Address: Dates:

http://jresearchbiology.com/ Received: 25 Mar 2013 Accepted: 02 May 2013 Published: 16 May 2013
documents/RA0344.pdf.

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.

940-946 | JRB | 2013 | Vol 3 | No 4

Journal of Research in Biology
An International
Scientific Research Journal www.jresearchbiology.com

INTRODUCTION
Many problems are opposed to a good
production of oil palm. These include pests, pathogenic
fungi and especially the continuing decline of pollinating
insects. For several years many problems of fruit set
were observed in some regions of cultivation of oil palm
tree causing a gradual decline in seed production (Mariau
et al., 1991). The observation was made that the most
affected areas, spread over large areas of lowland.
There are many evidences that the pollinator
insects effectively contributes to the reproduction of
many cultivated plant species. Regarding the oil palm
tree, the discovery of the role of insects in pollination
was made by Chevalier in 1910. The works of Syed
(1979) and Syed et al. (1982) have confirmed this
finding. Pollination of oil palm is essentially
entomophilous (Corrado, 1985). Without pollination,
fruit set by wind is extremely low (Mariau et al., 1991).
Pollinating insects are thus an undeniable role. In Cte
d'Ivoire (West Africa), five species of Elaeidobius
(E. kamerunicus, E. plagiatus, E. subvitatus,
E. bilineatus, E. singularis), two species of Microporum
(M. congolense and M. dispar), two species of
Proseostus (P. minor, P. sculptilis), Atheta burgeoni,
Gabrius sp, Litargus sp, Thrips sp and Anthocoris sp
have been described by Desmier De Chenon, (1981) and
Hala et al., (2012) as pollinators of oil palm. It therefore
seemed appropriate to follow the dynamics of these
insects on two different ecological zones: lowland and
plain. The research question that we asked is whether the
growing area of oil palm has an influence on populations
of pollinating insects. Many factors can explain the
fluctuations of insect populations. The latest studies on
this subject have established outside the bioclimatic
factors which have a clear implication, that the use of
insecticides in the fight against pests do not spare
beneficial insects that are pollinators (Tuo et al., 2011).
This preliminary study established a quantitative
and qualitative comparison of oil palm inflorescences
941
Koua et al., 2013
insects in two agro ecosystems: lowland and plain.
MATERIAL AND METHODS
Study site
Our study site was the experimental station of La
M, located at 526, N, 350, W. The station is located to
about thirty kilometers north-east of Abidjan (Cte
dIvoire). This area is characterized by an ombrophilous
forest (Traor and Mangara, 2009).
The study area has an equatorial climate
characterized by two distinct rainy seasons (March to
July and November). These two seasons are alternated by
two dry seasons: December to February and from August
to October (Pene and Assa, 2003). The monthly mean
temperature was about 27C. The monthly average of the
highest temperature was recorded in March and the
lowest in August with respectively 28.55 and 25.5C.
The average annual rainfall was about 1500 mm. The
average annual sunshine duration was about 1790 h; the
average monthly relative humidity was about 81%.
Insects of male inflorescences
Three operations were performed to assess the
male inflorescences of oil palm insect fauna: location,
sampling and identification of insects (Fataye, 1984).
Location
Each month a location was carried out in order to
count four inflorescences in the process of flowering.
This location has identified 192 male inflorescences at a
rate of eight per month during two years.
Sampling
As soon as the third florets of each inflorescence
were listed, with pair of secateurs four spikelets per
inflorescence were collected. This collection was done in
the beginning of anthesis (BA). The second (full
anthesis: FA) and third (end anthesis: EA) took place
respectively after the first three days and two days after
the second. Each batch of four spikelets collected was
placed in a bag and then the insects collected were
neutralized with an insecticide bomb. Before
Journal of Research in Biology (2013) 3(4): 940-946
Koua et al., 2013
identification, insects of each batch of spikelet were
collected in pillboxes containing alcohol 70%.
Identification
Using the collection of the insect fauna of oil
palm inflorescences of the National Agricultural
Research Centre of La ME and a binocular microscope,
insects of every month and each area were identified at
the species level.
Insects of female inflorescences
This study was conducted according to the
methods of N'goran (1982) and Fataye (1984).
Location
Two non-flowering inflorescences per plot were
identified and followed by month. The inflorescences
were cleared of husks and bulky leaves with machetes
and knives three days before flowering. Each
inflorescence thus revealed was covered with a muslin
cage and attached to the floral stem with a rubber. The
bagged inflorescences were controlled each afternoon to
follow the evolution of the inflorescence.
Sampling
All the insects that were attracted are placed on
the cage once flowering commences. Using a vacuum
cleaner, these insects were captured every hour for ten
minutes. This operation was performed at 6 AM to 6 PM
during the two days of the anthesis length. The collected
insects were immediately stored in pillboxes containing
70% alcohol. At the end of the day, insects collected
were sent to the laboratory. At each study site, sampling
was conducted on 48 inflorescences.
Identification
Insects collected were identified using the same
protocol as previously.
STATISTICAL ANALYSIS
Data processing was performed using Statistica
software version 7.1. An analysis of variance (ANOVA)
revealed significant differences between the data. The
test of Student-Newman-Keuls at 5% was used to
Journal of Research in Biology (2013) 3(4): 940-946
classify the means into homogeneous groups
RESULTS AND DISCUSSION
Variation in the number of insects on male
inflorescences
On male inflorescences of oil palm tree, the
insects mostly belong to Elaeidobius (E) genus. Five
species were observed: E. kamerunicus, E. plagiatus,
E. subvittatus, E. singularis and E. bilineatus.
Microporum (M) genus was represented by the species
M. dispar and M. congolense. Prosoestus genus was
present with two species P. sculptilis and P. minor.
Species, Atheta burgeoni, Lithargus sp., Anthocoride sp.,
Thrips sp, Gabrius sp. and bees (Nomia sp and
Apis mellifera) were also observed.
At the beginning of anthesis (BA)
The number of insects collected from the plot of
lowland (61%) is higher than that collected on the plain
(31%). At the species level, it was observed that
E. singularis, E. bilineatus, P. sculptilis, M congolense
and Anthocoris sp showed a significant difference
depending on the growing area with a higher effective in
lowland areas. The other species showed no preference
for one area (Figure 1A).
At full anthesis (FA)
The total number of insects collected was 42% in
the plain region and 58% in lowland areas. For the
species, E. plagiatus, E. kamerunicus, M. congolense,
M. dispar and A. Burgeoni, a significant difference was
found between their respective populations based on the
growing area. Only A. burgeoni presented a higher
effective in the lowland areas. The other species were
much more present in plain areas. Besides these species,
no differences were recorded between the number of
insects collected in lowland areas and those collected on
the plain (Figure 1B).
At the end of anthesis (EA)
The total number of insects differ from one area
to another. It was 75% in lowland areas against 25% on
942
 Koua et al., 2013

the plain. With regard to species, only the number of

E. singularis, M. dispar, Lithargus sp and A. burgeoni
depends on the area. These species excepted Lithargus sp
were more abundant in the lowland than on the plain
(Figure 1C).
Variation in the number of insects on female
inflorescences
The species observed on female inflorescences
A: Beginning of anthesis
were: E. kamerunicus, E. plagiatus, E. subvittatus,
E. bilineatus, E. singularis, M. congolense, M. dispar,
P. minor, P. sculptilis and Atheta burgeoni.
First day of anthesis
The total population of insects was significantly
higher on the plain (78%) than in the lowland area
(22%). At the species level, only the species M. dispar,
M. congolense, E. plagiatus, E. kamerunicus,
E. subvittatus, E. singularis, A. burgeoni and P. minor,
B: Full anthesis
were affected by the growing area. A. burgeoni attended
more inflorescences of the lowland area. Other species
were more present on the plain than in the lowlands
(Figure 2A).
Second day of anthesis
Insects were relatively influenced by the growing
area. Indeed, 78% of the insects were collected on
inflorescences of the plain region against 28% in the
lowlands. Regarding species, E. subvittatus, A. Burgeoni, C: End of anthesis
P. minor and P. sculptilis, had no preference for the Lowland Plain
growing area. The species E. kamerunicus, E. plagiatus, Histograms with the same letter are not
E. bilineatus, E. singularis, M. dispar and M. congolense significantly different at the 5% level
were receptive to the area of culture (Figure 2B). Figure 1: Influence of the growing area on the
Among the species, only E. kamerunicus, number of insects present on male inflorescences
E.su: Elaeidobius subvittatus; E.p: Elaeidobius
E. plagiatus, E. singularis and M. dispar were affected plagiatus; E.s: Elaeidobius singularis; E.b: Elaeidobius
by the growing area both the first and second day of balineatus; E.k: Elaiedobius kamerinucus;
P.m: Prosoestus minor; P.s: Prosoestus sculptilis;
anthesis. M.c: Microporum congolense; M.d: Microporum
The insects of oil palm inflorescences showed no dispar; L.sp: Lithargus sp.; Ant.sp.: Anthocoris sp.;
A.bur: Atheta burgeoni: T.sp.: Thrips sp.; G.sp.:
qualitative change from one area to another. These Gabrius sp.
species were always present on the oil palm tree and Sixteen insect species were observed on the male
colonize the inflorescences of this plant regardless of the inflorescences against only 10 species on female
study area. inflorescences. The six species that were absent on the
943 Journal of Research in Biology (2013) 3(4): 940-946
Koua et al., 2013

female inflorescences (Lithargus sp., Anthocoride sp.,

Thrips sp., Gabrius sp., and bees (Nomia sp. and
Apis mellifera) would not intervene mainly in oil palm
pollination. According to Mariau et al., 1991, four
species provide the largest share of pollination of oil
palm tree: E. kamerinucus, E. plagiatus, E. subvittatus
and E. singularis.
The male inflorescences showed a variation in
the numbers of insects based on the growing area and the
A: First day of anthesis
stage of flowering. In general, the lowland areas showed
significantly more insects than upland areas (61% against
31% at the beginning of anthesis, 58% against 42% at
full anthesis and finally 75% against 25% at the end of
anthesis). Insects observed in the male inflorescences
live on them usually. For example, the male flowers are
the breeding sites of insects of the genus Elaeidobius
(Beaudoin-Ollivier et al., 2012). The differences could
be explained by changes in environmental factors. At the
beginning of anthesis, flowers began to appear on the A: Second day of anthesis

male inflorescences that induced an attractive factor

Lowland Plain
because of the strong smell of anise emitted by the male
Histograms with the same letter are not
flowers. To this, were added the environmental factors significantly different at the 5% level.
including relative humidity and temperature. It was noted Figure 2: Influence of the growing area on the
that the number of insects on lowland inflorescences numbers of insects present on female inflorescences
E.su: Elaeidobius subvittatus; E.p: Elaeidobius
were two times higher than those of insects collected on plagiatus; E.s: Elaeidobius singularis; E.b: Elaeidobius
plain region inflorescences. At the species level, if for balineatus; E.k: Elaeidobius kamerinucus; P.m:
Prosoestus minor; P.s: Prosoestus sculptilis; M.c:
E. bilineatus, P. sculptilis and M. congolense, numbers Microporum congolense; M.d: Microporum dispar;
were highest in the lowland area than on the plain region. L.sp: Lithargus sp.; Ant.sp.: Anthocoris sp.; A.bur:
Atheta burgeoni: T.sp.: Thrips sp.; G.sp.: Gabrius sp.
These three species are unfortunately not effectively
intervening in the pollination of oil palm tree. The other M. congolense were significantly different from one area
insect species showed no significant difference at the to another. Indeed, apart A. burgeoni, all these species
beginning of anthesis. were more prevalent in upland areas. This can be
In full anthesis, the number of insects on male explained by bioclimatic factors which were more
inflorescences has reached its maximum value. The favorable to the activity of these insects.
attractive factor of flowers took over on bioclimatic We observed three times more insects on
factors. Thus, it has almost the same number of insects lowland inflorescences than on those on plain at the end
from one region to another. In terms of species observed of anthesis. This can be explained by the fact that on
in full anthesis, the number of E. plagiatus, plain, the flowers were fading and dry faster than the
E. kamerunicus, A. Burgeoni, M. dispar and inflorescences in lowland areas (lower temperature and

Journal of Research in Biology (2013) 3(4): 940-946 944


higher relative humidity). Insects were removed quickly
on inflorescences located in plain region. At the species
level, only E. singularis, M. dispar, A. burgeoni and
Lithargus sp. showed significantly different numbers
from one area to another. Only bio-ecological
requirements of these species can explain this
distribution. According to Hussein et al., 1990, the
change in the population of pollinating insects in
plantations of oil palm is largely due to the influence of
intrinsic and extrinsic factors, in particular, biological
and chemical factors.
During the anthesis, the total number of insects is
higher on female inflorescences taken from the plain
region than in the lowland area. At the species level, only
P. sculptilis showed no preference zone during the two
days during anthesis. The other species except
A. burgeoni showed a preference for the plateau region.
The determining factor is mainly the higher temperature
which allows the presence and maximal activity of
insects found on female inflorescences (Mariau et al.,
1991). Thus, the first day of anthesis as the second, the
numbers of insects were higher in these areas.
CONCLUSION
The number of insects collected on the plain
region is significantly different from that harvested the
lowland area. The number of insect has been higher in
male inflorescences in lowland areas than on the plain.
This number was higher on the plain than in the lowland
area. The numbers of insects are influenced by the
culture area. This factor is to be taken into account in the
implementation and the entomological monitoring of oil
palm plantations.
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Chevalier A. 1910. Documentation sur le palmier
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Fataye A. 1984. Rle des principaux insectes dans la
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Hala N, Tuo Y, Akpesse AAM, Koua HK and Tano
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Hussein MY, Lajis NH and Ali JH. 1990. Biological
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introduction of Elaeidobius kamerunicus faust, the oil
palm pollinator in Malaisia. Acta Horticulturae, 288: 81-
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Mariau D, Houssou M, Lecoustre R and Ndigui B.
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fleurs femelles des palmiers huile ; importance des
populations sur les fleurs mles. Rapport de stage de fin
de premire anne agronomique, ENSA, Abidjan-
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Station palmier huile IRHO-CIRAD de La M, CI.13p.

Syed RA. 1979. Studies on oil palm pollination by

insects. Bull. Ent. Res, 69(2): 213-224.

Syed RA, Law IH and Corley RH. 1982. VInsect

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Journal of Research in Biology (2013) 3(4): 940-946 946

 Journal of Research in Biology
Odonata diversity (Insecta: Arthropoda) in rice and vegetable fields in a
north-eastern district of Tamil Nadu, India
Journal of Research in Biology
Authors:
Veeramuthu Anbalagan,
Michael Gabriel Paulraj
and Savarimuthu
Ignacimuthu*
Institution:
Entomology Research
Institute, Loyola College,
Chennai-34.
Corresponding author: Keywords:
Savarimuthu Ignacimuthu Dragonflies, Damselflies, Libellulidae, Pantala flavescens
Email Id:
Web Address:
http://jresearchbiology.com/
documents/RA0346.pdf.
Journal of Research in Biology
An International
Scientific Research Journal
An International Scientific Research Journal
Original Research
ABSTRACT:
Odonata diversity in vegetable fields (brinjal and okra) and rice fields was
studied from January 2005 to December 2008 in Tiruvallur district of Tamil Nadu.
Totally 23 species of Anisoptera (dragonflies) and 12 species of Zygoptera (damselflies)
were recorded and all these species were grouped into eight families. In vegetable
fields 31 species of dragonflies and damselflies were recorded under 22 genera. In rice
fields the species richness (21 species) and total genera (16) were less than vegetable
fields during the entire study period. Libellulidae was the large family in both
vegetable and rice fields which comprised maximum number of species.
Pantala flavescens (Fabricius), a migratory species, was the most dominant in
numbers throughout the year. Diversity indices clearly showed that odonata diversity
was higher in vegetable fields than in rice fields.
Article Citation:
Veeramuthu Anbalagan, Michael Gabriel Paulraj and Savarimuthu Ignacimuthu.
Odonata diversity (Insecta: Arthropoda) in rice and vegetable fields in a north-eastern
district of Tamil Nadu, India.
Journal of Research in Biology (2013) 3(4): 977-983
Dates:
Received: 06 Apr 2013 Accepted: 23 May 2013 Published: 05 June 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.
977-983 | JRB | 2013 | Vol 3 | No 4
www.jresearchbiology.com

INTRODUCTION
Dragonflies and damselflies in the order Odonata
are important group of insects in agroecosystems, forest
ecosystems and aquatic ecosystems. They are potential
biocontrol agents of agricultural, horticultural and forest
pests. Many studies have shown that the larval stages of
Odonata are important biological control agents of
mosquito larvae (Mandal et al., 2008; Spencer et al.,
1999). According to Corbet (1999), dragonflies and
damselflies are excellent ecological indicators. Around
6,000 species and subspecies of Odonata have been
described under 630 genera in 28 families throughout the
world (Tsuda, 1991). In India, 499 species, 139 genera
and 17 families of dragonflies and damselflies have been
documented (Prasad and Varshney, 1995; Sharma,
2010). Odonata diversity has been extensively studied in
different forest areas. Emiliyamma (2005) has recorded
31 species of dragonflies and damselflies from southern
Western Ghats in the Kottayam district of Kerala. Very
few investigators have studied the Odonata diversity in
agricultural fields (Gunathilagaraj et al., 1999;
Kandibane et al., 2005). A knowledge on Odonata
diversity in different agro ecosystems is very essential to
understand the influence of crop type on species
richness, abundance and evenness of dragonflies and
damselflies. Hence the present work was undertaken to
assess the Odonata diversity in two different agricultural
fields, i.e. rice and vegetable fields in Tiruvallur district
of Tamil Nadu.
MATERIAL AND METHODS
Study site:
Dragonflies and damselflies were sampled from
vegetable fields, viz. brinjal and okra in Kolappancheri
village and rice fields in Vayalanallur village of
Tiruvallur district. The geocordination of Tiruvallur
district is 12 15 and 13 5`N Latitude and 99 15` and
80 20` E Longitude.
978
Anbalagan et al., 2013
Sampling of Odonates:
In each village, dragonflies and damselflies were
sampled in three different locations by quadrate method.
Quadrates of 25 m x 10 m size were laid down with
threads inside rice, brinjal and okra fields separately.
Totally three quadrates were put in each rice and
vegetable fields. Perched dragonflies and damselflies
found inside the quadrates were collected by sweeping
net (25 cm in diameter) during day times (between 10.00
AM to 15.00 PM). Flying Odonates inside quadrate area
were also caught with sweeping net. Sampling was done
twice in a month from January 2005 to December 2008.
Specimens from replications were pooled together.
Identification:
The specimens were identified using
identification keys provided by Fraser (1933, 1934 and
1936) and Subramanian (2009). After identification and
counting the total number of specimens, few specimens
from each taxa were retained and others were left behind
alive in the field. Specimens which were not identified in
the field were brought to the laboratory for identification.
The identified specimens were deposited at the
Entomology Research Institute, Loyola College,
Chennai.
Meteorological Data:
Data on atmospheric temperature, relative
humidity, mean total rainfall and total number of rainy
days from 2005 to 2008 were obtained from Regional
Meteorological Centre, Chennai.
Diversity indices:
Total number of dragonflies and damselflies
collected during the study period was recorded. Total
abundance, Simpsons index of diversity (1-D), Shannon
-Wiener Diversity Index (H), Shannon entropy, species
richness and species evenness were calculated by using
the software Past.exe (ver. 2.14). Jaccards similarity
index was calculated to find out the similarity in Odonata
diversity between vegetable and rice fields.
Journal of Research in Biology (2013) 3(4): 977-983
Anbalagan et al., 2013

The formulae for the diversity indices are as follows:

Simpsons index (D) = ni (ni-1) / N (N-1)
i=1
Where ni = number of individual for each species
N = total number of individuals

Shannon index of general diversity ( H )

ni ni
H =- N N
log
Where ni = number of individual for each species
N = total number of individuals
Figure 1. Total number of genera and species
Evenness (e) collected under different families of Odonata
collectively from vegetable and rice fields
H
e=
log S Coenagrionidae, Euphaeidae, Lestidae and
Platycnemididae were recorded under Zygoptera
(damselflies) (Table 1). Totally 31 species of dragonflies
Where H = Shannon index
and damselflies were recorded under 22 genera, of which
S = number of species
15 genera and 22 species were dragonflies and 7 genera
The similarity in odonata diversity between
and 9 species were damselflies. Libellulidae was found
vegetable fields and rice fields was assessed by using the
to be the largest family, which has the highest number of
formula of Jaccards similarity index as follows:
species (18 species) throughout the study period. Species
Jaccards Index = A/ (A+B+C)
richness was 31 throughout the study. Total abundance
Where A= total number of species present in both
was maximum (4167) in 2008. Maximum evenness of
communities
0.899 was recorded in vegetable fields in 2007 and this
B= the number of species present in community 1 but not 2
was correlated with the maximum Shannon-Wiener
C= the number of species present in community 2 but not 1
diversity index of 3.328 during the same study year
(Table 2). The similarity index (Jaccards similarity
RESULTS
index) was calculated as 0.660 for each study
Totally 35 species of dragonflies and damselflies
year (Table 2).
were recorded collectively from vegetable and rice fields
Species composition and diversity in rice fields
in Tiruvallur district from January 2005 to December
Five different families namely Aeshnidae,
2008 (Figure1). The species composition, richness,
Gomphidae, Libellulidae, Coenagrionidae and Lestidae
evenness and other diversity indices showed variations
were recorded in rice field. All the species collected from
between vegetable and rice fields.
rice fields were grouped under 16 genera (12 Anisoptera
Species composition and diversity in vegetable fields
and 4 Zygoptera). Total number of species recorded in
Three families viz., Aeshnidae, Gomphidae and
rice field was 21 (15 Anisoptera and 6 Zygoptera).
Libellulidae were recorded under Anisoptera
Maximum total abundance (1703) was recorded in 2008.
(dragonflies) and five families viz., Calopterygidae,
Maximum Shannon-Wiener diversity index (2.871) and

Journal of Research in Biology (2013) 3(4): 977-983 979

 Anbalagan et al., 2013

Table 1. Taxonomic composition and total number of individuals collected under different species of Odonata
from North-Eastern Tamilnadu during 2005-2008
Sl.No. Species Number of individuals collected
Vegetable fields Rice fields
2005 2006 2007 2008 2005 2006 2007 2008
Anisoptera
Family: Aeshnidae
1 Anax guttatus (Burmeister) 0 0 0 0 9 11 7 6
2 Anax immaculifrons (Rambur) 25 52 67 72 12 6 14 11
Family: Gomphidae
3 Heliogomphus selysi (Fraser) 179 158 124 186 38 88 76 54
4 Ictinogomphus distinctus (Rambur) 128 94 108 134 0 0 0 0
5 Ictinogomphus rapax (Rambur) 112 75 82 92 29 42 37 32
Family: Libellulidae
6 Brachythemis chalybea (Brauer) 128 142 108 129 0 0 0 0
7 Brachythemis contaminata (Fabricius) 106 85 122 148 78 55 86 73
8 Bradinopyga geminata (Rambur) 27 35 42 33 0 0 0 0
9 Crocothemis servilia (Drury) 220 145 189 238 36 42 46 58
10 Diplocodes trivialis (Rambur) 175 205 218 232 125 163 158 182
11 Neurothemis tullia (Drury) 98 112 147 121 58 82 117 93
12 Orthetrum glaucum (Brauer) 116 105 98 165 78 67 63 85
13 Orthetrum sabina (Drury) 125 145 102 148 51 25 48 60
14 Orthetrum testaceum (Burmeister) 114 108 122 148 0 0 0 0
15 Pantala falvescens (Fabricius) 480 306 318 372 185 211 203 197
16 Rhyothemis variegata (Linn.) 219 184 225 236 89 58 62 71
17 Sympetrum vulgatum flavum (Bartenef) 90 109 128 114 0 0 0 0
18 Tholymis tillarga (Fabricius) 30 18 45 55 0 0 0 0
19 Tramea basilaris (Palisot de Beauvois) 170 165 138 145 31 27 41 29
20 Tramea limbata (Desjardins) 150 120 111 165 0 0 0 0
21 Trithemis aurora (Burmeister) 112 78 65 92 26 34 31 40
22 Trithemis festiva (Rambur) 107 118 128 108 0 0 0 0
23 Trithemis pallidinervis (Kirby) 72 110 95 108 52 45 69 42
Zygoptera
Family:Calopterygidae
24 Caliphaea sp 27 35 42 33 0 0 0 0
Family: Coenagrionidae
25 Agriocnemis femina femina (Brauer) 0 0 0 0 110 74 101 122
26 Agriocnemis pygmaea (Rambur) 0 0 0 0 92 68 81 105
27 Ceriagrion coromandelianum(Fabricius) 190 78 158 212 140 61 125 156
28 Ischnura aurora (Brauer) 70 78 65 128 43 59 78 88
29 Ischnura delicata (Hagen) 0 0 0 0 121 82 88 106
30 Ischnura inarmata (Calvert) 71 65 108 108 0 0 0 0
31 Ischnura senegalensis (Rambur) 92 84 149 132 0 0 0 0
Family: Euphaeidae
32 Euphaea sp 30 45 68 73 0 0 0 0
Family: Lestidae
33 Lestes viridulus (Rambur) 69 80 118 120 61 68 91 93
Family: Platycnemididae
34 Copera marginipes (Rambur) 70 78 92 55 0 0 0 0
35 Platycnemis sp 54 65 88 65 0 0 0 0
Total 3656 3277 3670 4167 1464 1368 1622 1703

evenness (0.8409) in rice fields were recorded during similarity index) was calculated as 0.660 for each study
2007. Odonata diversity in rice fields was lower than year.
vegetable fields. The similarity index (Jaccards
980 Journal of Research in Biology (2013) 3(4): 977-983
Anbalagan et al., 2013

Table 2. Diversity indices for Odonata in vegetable and rice fields from 2005 to 2008
2005 2006 2007 2008
Sl.
Diversity Indices Vegetable Rice Vegetable Rice Vegetable Rice Vegetable
No. Rice Fields
Fields Fields Fields Fields Fields Fields Fields
1 Species richness (S) 21 31 21 31 21 31 21 31
Total no. of
2 1464 3656 1368 3277 1622 3670 1703 4167
individuals
Shannon-Wiener
3 2.84 3.221 2.828 3.3 2.871 3.328 2.847 3.308
Diversity Index (H)
4 Simpson 1-D 0.9326 0.9508 0.9288 0.9584 0.9358 0.9601 0.934 0.9591
5 Evenness 0.815 0.8082 0.8056 0.8749 0.8409 0.8991 0.8208 0.8817
Jaccard Similarity
6 0.660 0.660 0.660 0.660
Index

Table 3. Mateorological data for the years from 2005 to 2008

Year Mean Maximum Mean minimum Mean Relative Mean Total Total number of
temperature (oC) temperature (oC) Humidity (%) @ Rainfall (mm) rainy days (2.5mm
0830/1730 hrs IST and above)
2005 33.6 24.8 66.8-75.6 199.8 73
2006 33.8 24.6 64.3-75.9 123.9 67
2007 33.3 24.5 67.1-75.7 106.9 68
2008 33.7 24.8 64.3-75.3 150.2 63
Meteorological data: Several investigators have reported that
The meteorological data is given in the table 3. dragonflies and damselflies are very common in rice
Mean maximum and minimum yearly temperatures were agroecosystems. Kandibane et al., (2003) have recorded
low in 2007 compared to other three years. Also the 12 species of Odonata under three families in rice fields
relative humidity was high in the year 2007. of Madura. In the present work the number of species
and families recorded in rice fields were high compared
DISCUSSION to the results of Kandibane et al., (2003, 2005). Among
Present study reports the odonata diversity in the various species, Pantala flavescens, a migratory
vegetable and rice agroecosystems. Odonates are species, was abundant in numbers. The damselfly
predaceous insects and they are important biocontrol Ceriagrion coromandelianum was abundant in both
agents of agricultural pests and vector mosquitoes. vegetable fields and rice fields. In rice field,
In the present study families Libellulidae in Anisoptera Agriocnemis femina femina was also found to be
and Coenagrionidae in Zygoptera were found to be more abundant. Kandibane et al., (2003) have reported that
diverse families in terms of the number of species. A. femina was more abundant in rice ecosystems.
Similar findings were already reported by some The diversity and distribution of insects may be
investigators. Ghahari et al.,(2009) have reported that influenced by type of ecosystems and climate. In the
families Libellulidae and Coenagrionidae were dominant present study the species richness, total abundance and
in terms of number of species in rice fields in Iran. diversity of Odonata were high in vegetable ecosystems
Kumar and Mitra (1998) reported that family compared to rice ecosystem. Higher evenness values
Libellulidae was represented by high number of species were recorded in vegetable fields than rice fields during
(18 species) among a total collection of 42 species from 2006, 2007 and 2008. When the richness and the
Sahstradhara, Dehra Dun. Similar reports were evenness of a community increases, the Shannon index
published by Prasad (2002), Kumar (2002) and also increases. In the present study the Shannon index
Vashishth et al., (2002). was higher in vegetable fields than rice fields. This was
Journal of Research in Biology (2013) 3(4): 977-983 981

due to the higher species richness and evenness in
vegetable fields. The dominance of species was found to
be lower in vegetable crops compared to rice fields.
Hence the Simpsons index of diversity (1-D) was higher
in vegetable crops and it clearly explained that species
distribution in vegetable crops was equal.
Besides the type of crop, the climatic factors
such as rainfall, atmospheric temperature and humidity
also affect the insect diversity. The average annual
temperature was the lowest in the year 2007. This lowest
average temperature in 2007 coincided with the
maximum insect diversity in both rice and vegetable
crops. Brinjal and okra plants grow taller with branches
and provide suitable microclimate and resting place for
perching adult Odonata. Vegetable fields also harbour
variety of small insects, which are the main prey of
Odonates. Latif et al., (2009) have reported 20 species
of pest insects and 10 families of predaceous insects in
brinjal field. Hence the presence of variety of prey
insects might be the reason for higher odonata diversity
in vegetable fields.
CONCLUSION
It is concluded that dragonfly and damselfly
diversity was influenced by type of crop because
vegetable ecosystem supported more taxa of Odonates
than rice field.
ACKNOWLEDGMENTS
The authors thank Entomology Research Institute
for financial support
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Cornell University Press, Ithaca, New York, 829.
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Ghahari H, Tabari M, Sakenin H, Ostovan H and
Imani S. 2009. Odonata (Insecta) from Northern Iran,
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Gunathilagaraj K, Soundarajan RP, Chitra N and
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Kandibane M, Mahadevan NR and Gunathilagaraj
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Kumar A. 2002. Odonata Diversity in Jharkhand State
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297-314.
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MM. 2009. Survey of arthropod biodiversity in the
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 Journal of Research in Biology An International Scientific Research Journal

Original Research

A chromosomal analysis of seven Cameroonian Acrididae species (Orthoptera:

Acridinae, Oedipodinae and Spathosterninae) based on published data
Journal of Research in Biology

Authors: ABSTRACT:
Seino Richard Akwanjoh1,2
and Dongmo Tonleu Ingrid1. So far, the karyotypes of seven Acrididae species from Cameroon have been
reported. These species included: Acrida turrita, Chirista compta, Coryphosima
stenoptera producta, Oxycatantops spissus (Acridinae), Paracinema luculenta,
Morphacris fasciata (Oedipodinae) and Spathosternum pygmaeum (Spathosterninae).
Institution:
1. Laboratory of Applied Karyotype and meiosis relationships among these species were analysed from
Ecology (LABEA), published data. The species had a common karyotype made up of 23 acrocentric
Department of Animal chromosomes (males), the sex mechanism in all seven species was XX-XO
Biology, Faculty of Science, and meiosis was normal and chiasmate. The chromosomes in the species occurred in
University of Dschang, three size groups of long, medium and short. The number of chromosomes per size
P.O. Box 353, Dschang, group however varied among the species (A. turrita = 4LL:5MM:2SS;
Cameroon. C. compta =4LL:4MM:3SS; C. stenoptera product=2LL:6MM:3SS; O. spissus
=5LL:3MM:3SS; P. luculenta = 6LL:2MM:3SS; M. fasciata = 6LL:2MM:3SS; and
2. Department of Biological S. pygmaeum = 2LL:7MM:2SS). The X chromosome was long in the Oedipodinae,
Science, Faculty of Science, medium in the Acridinae and short in the Spathosterninae. Total length of
University of Bamenda, chromosomal material was in the series C. compta > O. spissus > P. luculenta >
P.O. Box 39, Bamenda, S. pygmaeum > A. turrita > M. fasciata > C.s. producta.
Cameroon.

Corresponding author: Keywords:

Seino Richard Akwanjoh Acrididae, Acridinae, Oedipodinae, Spathosterninae, karyotype, relationships.

Email: Article Citation:

raseino@yahoo.co.uk Seino Richard Akwanjoh and Dongmo Tonleu Ingrid.
A chromosomal analysis of seven Cameroonian Acrididae species (Orthoptera:
Acridinae, Oedipodinae and Spathosterninae) based on published data.
Journal of Research in Biology (2013) 3(4): 947-953

Web Address: Dates:

http://jresearchbiology.com/
Received: 15 Apr 2013 Accepted: 02 May 2013 Published: 20 May 2013
documents/RA0349.pdf.

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.

947-953 | JRB | 2013 | Vol 3 | No 4

Journal of Research in Biology
An International Scientific
Research Journal www.jresearchbiology.com
 Seino and Dongmo, 2013

INTRODUCTION Coryphosima stenoptera producta, Oxycatantops spissus

The use of Orthoptera material for karyotype
studies dates from the inception of cytogenetics. This is
simply because Orthoptera material presents large
chromosomes and few chromosomes per karyotype.
Chromosome size and number are of important
cytotaxonomic value (Turkoglu and Koca, 2002). The
Orthoptera are also well known for their karyotypic
uniformity in chromosome number and morphology
(Ashwathanarayana and Ashwath, 2006; Chadha and
Mehta, 2011a).
It has been severally shown that analysis of
karyotype differentiation between species yields better
understanding of the evolutionary interrelationships and
divergence (Chadha and Mehta, 2011a; Sandhu and
Chadha, 2012). A survey of investigations on karyotype
evolution in different groups of animals has revealed that
several karyotypes are dynamic and are subject to
change. Therefore, the stable karyotypes of the Acrididae
are subject to change.
The cytogenetic diversity of Cameroonian
acridid grasshoppers has not been investigated. During
this study, published data on karyotypic characters were
analysed to determine similarities and differences as well
as interrelationships among seven Cameroonian
Acrididae species.
MATERIALS AND METHODS
The cytogenetics of only seven Cameroonian
Acrididae species have so far been described. The
species include Acrida turrita, Chirista compta,
(Sub-family Acridinae) Paracinema luculenta,
Morphacris fasciata (Sub-family Oedipodinae) and
Spathosternum pygmaeum (Sub-family Spathosterninae).
The species and the sources from which karyotypic
information on them was obtained for this analysis are
shown in Table 1.
To analyse these karyotypes for similarities and
differences, the karyotypes of the seven species were
also arranged together (Figure. 1) and the morphometric
characters for the seven species were arranged in a
tabular form (Table 2).
RESULTS
Information on chromosome number,
morphology, size, and length of X chromosome obtained
for the seven species is summarised in Table 2. A perusal
of Table 2 revealed that among the seven species studied
A. turrita, C. compta, C. stenoptera producta and
O. spissus belonged to the sub-family Acridinae,
P. luculenta and M. faciata belonged to the subfamily
Oedipodinae and S. pygmaeum belonged to the
subfamily Spathosterninae (Mestre and Chiffaud, 2009).
Table 2 also revealed that the seven species investigated
had a common a diploid chromosome number of 2n=23
and the sex determining mechanism was XO in males.
Figure 1 also revealed that the in the seven species
investigated was acrocentric in morphology. The
chromosomes in all seven species occurred in three size
groups of long, medium and short. The number of
chromosome pairs per size group varied between species

Table 1: The species analysed, their subfamilies and references from which karyotypic
information was obtained
S/No Species Subfamily Source of data
1 Acrida turrita Seino et al, 2008
2 Chirista compta Seino et al, 2010
3 Coryphosima stenoptera producta Acridinae Seino et al, 2010
4 Oxycatantops spissus Seino et al, 2010
5 Paracinema luculenta Seino et al, 2012
6 Morphacris fasciata Oedipodinae Seino et al, 2012
7 Spathosternum pygmaeum Spathosterninae Seino et al, 2012
948 Journal of Research in Biology (2013) 3(4): 947-953
 Seino and Dongmo, 2013

and subfamilies (Table 2; Figure. 2). The Oedipodinae

chromosome
showed most similarity since both of them revealed

Nature of X
6 long, 2 medium and 3 short chromosomes (6LL: 2MM:
3SS) in their karyotypes. The lengths of the

S
X chromosome was in the series P. luculenta >
chromosome
(m ) of X

C. compta > O. spissus > C.s. producta > M; fasciata >

5.00.08

7.30.52

5.60.56

6.60.00

7.70.59

5.50.24

1.70.00
Length

A. turrita > S. pygmaeum. However, the X chromosome

was medium in the Acridinae, long in the Oedipodinae
All

All

All

All

All

All

All
A

and short in the Spathosterninae species (Figure. 2). The

Morphology of
chromosomes

total length of chromosomal material was in the series

Table 2: Morphometric characters of karyotypes of the seven species investigated

SM

-
C. compta> O. spissus> P. luculenta> S. pygmaeum>
M

A. turrita> M. fasciata > C.s. producta.

-

-
chromosome
length (m)

134.60.79

176.30.14

123.67.60

165.40.61

164.72.26

129.90.63

160.70.91

DISCUSSION
Total

Every species has a unique karyotype which

provides an identity to the species (Channaveerappa and
Short

Ranganath, 1997). Acridid grasshoppers are known to be

Number of chromosome per

characterised by a basic karyotype of 23 chromosomes in

M= Metacentric, SM= Submetacentric, A= Acrocentric; L=long, M=Medium, S=Short

males. Due to this great cytogenetic uniformity Acridids

Medium

are considered as an example of karyotypic

5

7
size group

conservation (Aswathanarayana and Aswath, 2006).

Long

In the present study, seven Acridids have been

4

investigated which belong to three different sub-families

mechanism

that include the Acridinae, Oedopodinae and

Sex deter-

XX-XO

XX-XO

XX-XO

XX-XO

XX-XO

XX-XO

XX-XO
mining

Spathosterninae. The results of this study revealed that

-

the seven Acrididae have a chromosome number of

per cell in the

23 and a sex determining mechanism which is XO/XX.

chromosomes
number of

Similar observations have been made for several other

Total

male

Acrididae species. With respect to chromosome number,

23

23

23

23

23

23

23

chromosome morphology and sex determining

mechanism, Bugrov et al., (1994); Bugrov (1995),
Spathosternina
Oedipodinae

Oedipodinae
Sub- family

Acridinae

Acridinae

Acridinae

Acridinae

Bugrov et al., (1999) Bugrov and Sergeev (1997)

observed similar results for Podisma and
Eyprepocnemidinae (Acrididae) grasshoppers in Russia
C. S. producta

S. pygmaeum

and Central Asia. Camacho and Cabrero (1983) also

P. luculenta

M. fasciata
C. compta

O. spissus
A. turrita

reported similar results for European species of Acrotylus

Species

(Oedopodinae). Yao (2006) and Chadha and

Mehta (2011a) reported similar results for
S.N

Spathosternum pransiniferum (Spathosterninae)

Journal of Research in Biology (2013) 3(4): 947-953 949

 Seino and Dongmo, 2013

a b c

d e f

g
Figure. 1: Mitotic Metaphase chromosomes in the seven species investigated.
a) Acrida turrita, b) Chirista compta, c) Coryphosima stenoptera producta, d) Oxycatantops spissus,
e) Paracinema luculenta, f) Mophacris fasciata, g) Spathosternum pygmaeum. Chromosomes are tapered
towards one end and centromeres were deemed to be towards the tapered ends of the chromosomes.

respectively from Asia and India. So the Acridid Acrididae species (White, 1973; Sharma and Gautam,
grasshoppers of different regions are showing 2002; Mayya et al., 2004; Chadha and Mehta, 2011a).
cytogenetic uniformity regarding chromosome number, Turkoglu and Koca (2002) reported the presence of
morphology and sex determining mechanism. The results metacentric, submetacentric and acrocentric
of this study confirmed that the basic Acrididae chromosomes in the karyotypes of Oedipoda schochi and
karyotype is 23 acrocentric chromosomes and a sex Acrotylus insbricus (Oedopodinae) from Turkey. The
determining mechanism of the XX/XO type. aberrant chromosomes were the result of centric fissions.
Metacentric chromosomes through fusions were not X - autosome fusion resulting in the Neo - XY sex
observed in the seven species here investigated even mechanism have been reported in some acridid
though they have been reported in several other grasshoppers (White, 1973). Bidau and Marti (2000)
950 Journal of Research in Biology (2013) 3(4): 947-953
Seino and Dongmo, 2013

16 16

Relative Chromosome Length

Relative Chromosome Length

14 14
12 12
10 10
(RCL)

(RCL)
8 8
6 6
4 4
2 2
0 0
L1 L2 L3 L4 M5 M6 M7 M8 M9 S10 S11 X L1 L2 L3 L4 M5 M6 M7 M8 S9 S10 S11 X
Chromosome pair Chromosome pair

A. turrita C. compta

18 16
Relative Chromosome Length

Relative Chromosome Length

16 14
14 12
12
10
10
(RCL)

(RCL)
8
8
6
6
4 4

2 2
0 0
L1 L2 M3 M4 M5 M6 M7 M8 S9 S10 S11 X L1 L2 L3 L4 L5 M6 M7 M8 S9 S10 S11 X
Chromosome pair Chromosome pair

C.s.producta O. spissus

14 14
Relative Chromosome Length
Relative Chromosome Length

12 12

10 10

8 8
(RCL)
(RCL)

6 6

4 4

2 2

0 0
L1 L2 L3 L4 L5 L6 M7 M8 S9 S10 S11 X L1 L2 L3 L4 L5 L6 M7 M8 S9 S10 S11 X

Chromosome pair Chromosome pair

P. luculenta M. fasciata
16
Relative Chromosome Length

14
12
10
(RCL)

8
6
4
2
0
L1 L2 M3 M4 M5 M6 M7 M8 M9 S10 S11 X
Chromosome pair

S. pygmaeum
Figure. 2: Idiograms of the seven species investigated
Journal of Research in Biology (2013) 3(4): 947-953 951

r eport ed Neo-XY in Dichropl us v ittatus
(Acrididae: Melanoplinae). This type of sex
determination mechanism was absent in the seven
species investigated in this study.
The X-chromosome during this investigation was
found to be medium in the four Acridinae. However,
Chadha and Mehta (2011a), investigating Indian
Acridinae observed that the X chromosome in A. turrita
was the longest chromosome in the karyotype. There is
therefore disagreement of this report with that of the
present investigation. Chadha and Mehta (2011b)
reported the X chromosome in Oedaleus abruptus
(oedipodinae) to be the largest element in the karyotype.
During the present study, the X-chromosome in
P. luculenta and M. fasciata (Oedipodinae) were among
the large chromosomes of the karyotypes. There is no
doubt that the X chromosomes of different species of the
Oedipodinae is one of the largest elements in the
karyotype. Though this chromosome was acrocentric in
the two Oedipodinae investigated here, Turkoglu and
Koca (2002) found the same chromosome in Oedipodia
schochi schochi and Acrotylus insbricus (Oedipodinae)
from Turkey to be Metacentric in morphology.
ACKNOWLEDGEMENTS
The authors are thankful to Dr Watcho Pierre
(Associate Professor in the Department of Animal
Biology, University of Dschang - Cameroon) for reading
the manuscript and helpful suggestions. We are also
grateful to Professor Mpoame Mbida (Head of the
Laboratory of Applied Ecology (LABEA), University of
Dschang for laboratory space.
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