Bibha Chetia Borah

Indira Baruah

Sonmoina Bhuyan

U C Goswami

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Save Nature to Survive
8(3): 1111-1114, 2013 (Supplement on Toxicology)
www.thebioscan.in
1111
EFFECT OF SUBLETHAL CONCENTRATION OF MALATHION ON
HEMATOLOGICAL PARAMETERS OF MONOPTERUS CUCHIA
(HAMILTON- BUCHANAN)
INDIRA BARUAH1*,  U. C. GOSWAMI2,  BIBHA CHETIA BORAH3,  SONMOINA BHUYAN3
1Department of Zoology, Bahona College, Jorhat
2 Department of Zoology, Gauhati University, Guwahati
3Fisheries Research Centre, Assam Agricultural University, Jorhat - 785 013
e-mail: indira_baruah@rediffmail.com
INTRODUCTION
Extensive use of chemicals is in vogue in present day
agriculture for enhancing crop productivity per unit area to
meet the demand of food for the growing population of the
world, particularly in developing countries like India. Among
the different chemicals used in modern agricultural systems,
pesticides are unique toxic substances, as these are deliberately
applied or added to suppress or eliminate some components
of the food chain of crop field ecosystem, commonly known
as pests, to safeguard the crop from being damaged or feed
upon. However, while doing so, the toxic chemicals affect
other non target components of the ecosystem and pollute
the soil, air and water. More so, when the pesticides enter the
aquatic ecosystem through direct application or run off from
agricultural operation they often cause adverse impact on the
aquatic biota including fish (Kumar, 1994). One of the very
commonly used pesticides in India is ‘malathion’ an
organophosphorus pesticide. The toxic effects of the organo
phosphorus components are the results of their ability to inhibit
the enzyme Acetyl chlorine esterase (ACHE) which in turn
disrupts the neural transmission through binding of the ACHE
with ‘Malatoxin’, the oxygen analog of Malathion (O’Brian et
al., 1974). Fish species, inhabiting the paddy fields, swamps,
canals and derelicts that are directly linked with paddy field
are more susceptible to exposure to such chemicals. Fresh
water eel Monopterus cuchia (Ham Buch) commonly known
as “Mud eel” is one such species which needs immediate
ABSTRACT
Impact of pesticides used in agricultural operations on the environment as well biotic community as a whole is
a global concern particularly in developing countries like India. The present study encompasses impact of sub
lethal concentration of a commonly used OP pesticide ‘Malathion’ on certain hematological parameters of
Monopterus cuchia (Ham- Buch) popularly known as ‘mud eel’ under laboratory condition. The study revealed
significant decrease in Total Erythrocyte Count (TEC) in treated fish upon exposure to sub lethal dose of Malathion
with a mean value of (2.02 ± 0.2) 106/mm3 in comparison to the control (2.49 ± 0.2) 106/mm3. In addition to
that, significant increase in leucocytes count in treated fish with a mean value of 2.57 ± 0.5 106/mm3 in
comparison to control (2.00 ± 0.5) 106/mm3 indicated ailing condition of the fish. Significant difference in
percentage of Eosinophil, Neutrophils, Lymphocytes, Monocytes in blood samples of controlled and treated fish
indicates the enormity of the impact of the pesticide on the hematological parameters of the fish.  Although
immediate mortality was not recorded, the findings indicated that prolonged exposure may have severe impact on
the life processes of the species.
KEYWORDS
Malathion
Hematological
parameter
IUCN
Metabolism
Received on :
17.05.2013
Accepted on :
26.08.2013
*Corresponding
author
attention in this aspect. Earlier study conducted by Baruah et
al. (2008) on behavioral response of the fish to different
concentration of the pesticides indicated that this species is
highly sensitive to exposure to malathion even in very low
concentration leading to weak and ailing health condition.
Hematological parameters reflect health status of fish and have
been used for detecting disease, dietary deficiency and
environmental stress (Conroy 1972; Srivastava, 1979). A study
of hematological parameter is therefore necessary to assess
the extent of damage caused by any toxic material. M cuchia
has very high demand among the ethnic population of the NE
region of India for its unique taste and therapeutic value and
is collected from its natural habitats like beels, swamps and
paddy fields by traditional method. The species have this
special nature of burrowing in mud throughout its life. The
natural population of this species is apparently on a decreasing
trend during the recent years.  The conservational status of
the species is reported to be lower risk near threatened as per
IUCN based CAMP report (1998). Increasing demand for the
species has been observed during recent years as a major
component of diet of anemic and weak people for the popular
belief that consumption of this species increase hemoglobin
count of blood and enhance physical strength in ailing person.
In addition to its importance as food fish, the species is known
to have ornamental value for its peculiar snakelike body shape
and movement. The species is enlisted as one of the 58 fish
species currently being exported as ornamental fish species
from NE Region of India. Vulnerability of this species to pesticide
1112
INDIRA  BARUAH  et  al.,
exposure due to its habits of mud dwelling as well as habit of
living in paddy field swamps etc., it is important to assess the
impact of different pesticides on the health and population
status of the species. In view of the above present study was
conducted on the toxic affect of malathion on some
hematological parameters like total blood cell count,
Differential Count of leucocytes (DLC)of M. cuchia, under
exposure to sub lethal concentration of Malathion.
MATERIALS AND METHODS
M. cuchia of average length (60-68 cm) and weight (180-250
gm), procured from the local wet-lands of Bahona area, Jorhat,
Assam were used during this study as test animals. The fish
were acclimatized to the controlled condition for 5 days period
to treatment. The concentration of malathion 50 EC is kept at
sublethal level (0.006 ml/l) determined on the basis of bioassay
trial. The test fishes (Total no. 30) were kept in laboratory
conditions in 2 glass aquaria separately each filled with 10
liters pond water treated with 0.006 ml malathion 50 EC/l
water for 25 days. One group of test fish (10 no.) placed in
fresh water without pesticide served as the control group. The
acclimatized fishes were not fed during the course of the
experiment (Delala et al., 1980). The pond water was changed
on alternate days and the concentration of pesticide was
maintained. In the treated and control fishes, the blood was
collected from the lateral line of experimental fishes by inserting
a heparinized microsyringe. The following methods were used
for the different blood parameters.
I. Total Erythrocyte count (RBC, 106/mm3): The total
erythrocytes count was estimated by the dilution chamber
technique using Hayem’s fluid as diluent (Benjamin,
1985).
II. Total leucocytes count (WBC 103/mm3): Total leucocytes
count was estimated by the standard dilution technique
using diluting fluid (4% glacial acetic acid and two drops
of gentian) (Talib and Khurana, 1995).
III. Differential count of leucocytes (DLC): Differential count
of blood smears prepared by staining with Giemsa stain
were made under 100X by using, and counting was done
by the Meander method. Hundred cells were counted
and tabulated viz. Neutrophils, Eosinophils, Basophils,
lymphocytes and monocytes.
IV. The total number of differential leucocytes present was
calculated as per the formula. DLC = Number of types
cells/Total number of white blood corpuscles × 100
V. Haemoglobin (Hb%): Haemoglobin was estimated by
Shalis and Haematin method as per modified by Talib
and Khurana (1995).
RESULTS AND DISCUSSION
Treatment with sub lethal concentration of Malathion (0.006
ml/l) has not indicated any immediate behavioral response as
well as mortality. However, the fish become lethargic and
subsequently become still upon exposure to prolonged period.
Mortality has not been recorded in any of the treatments during
the period of investigation.
Hematological investigations carried out by taking blood
samples from control and treated fishes revealed that the Total
Erythrocyte Count (TEC) had been reduced considerably in
treated specimen with an average TEC ranging from (1.00 ±
0.3)106/mm3 to (2.00 ± 0.2) 106/mm3 and a mean 2.02 ± 0.2
106/mm3 while in the control specimen it was (2.20 ± 0.5)106/
mm3 to (3.20 ± 0.5) 106/mm3 and a mean 2.49 ± 0.2 106/
mm3. The difference was significant at 1% level. On the other
hand there was significant increase in Total Leukocyte Count
(TLC) (Table 1). Significant differences in the TEC and TLC
records of treated and controlled group indicated severity of
Table 1: Impact of malathion (0.006 mL/L) on the different blood parameters of Monopterus cuchia
Group No. Fish group treatment Normal control TEC 106/mm3 TLC 103/mm3 Hb%
and No. of fish period (days) period (days) Control Treated Control Treated Control Treated
1 (n=8) 5 5 3.20±5 2.00±0.2 2.80±0.2 2.00±0.1 18±4 17.0±0.5
2 (n=8) 10 10 2.60±0.3 1.60±0.5 2.00±0.5 1.80±0.5 16±5.5 12.5±1.5
3 (n=8) 15 15 2.25±0.3 1.00±0.3 1.80±0.1 1.50±0.5 12±2.5 9.2±2.5
4 (n=5) 20 20 2.20±0.5 1.50±0.3 1.80±0.1 1.55±0.5 10±1.5 7.0±3.0
5 (n=5) 25 25 2.20±0.5 1.60±0.4 1.60±.05 1.00±0.5 8±0.5 5.5±0.5
Mean 2.49±0.2 2.02±0.2 2.00±.05 2.57±0.5 12.8±1.5 12.64±1.5
‘t’ - value between control and treated 5.875** 4.924** 0.266NS
Table 2: Effect of malathion (0.006 mL/L) on the DLC of Monopterus cuchia
Group Fish group Normal Eosinophil % Neutrophils % Basophils % Lymphocytes % Monocytes %
No. and treatment control Control Treated Control Treated Control Treated Control Treated Control Treated
No. of period period
fish (days) (days)
1 (n=8) 5 5 4±0.5 3±0.5 45±5.5 36±6.0 2±0.5 2±0.5 55±7.5 60±10.0 19±2.0 27±4.5
2 (n=8) 10 10 4±1.2 3±0.5 35±7.0 25±4.0 1.5±1.5 2±0.5 47±2.5 58±5.0 16±1.5 25±3.5
3 (n=8) 15 15 3±0.1 2±1.0 31±8.0 22±3.0 1±1.5 1±0.5 45±3.0 53±3.5 20±1.5 27±2.5
4 (n=8) 20 20 2±0.5 1±0.2 32±9.0 22±2.0 1±1.5 1±0.5 50±9.5 58±3.5 18±1.5 23±3.5
5 (n=8) 25 25 2±0.5 1±0.1 30±5.0 23±1.5 1±1.5 1±0.5 52±0.11 55±2.5 18±2.5 20±4.5
Mean 3±0.2 1±0.01 34.6±5.5 25.6±0.2 1±1.5 1.2±3.0 49.8±7.5 56.8±0.7 18.2±2.5 24.4±3.0
‘t’ - value between 3.207** 16.432** 0.535NS 5.078** 4.966*
control and treated
* - Significant at 5% level of significance,  ** - Significant at 1% level of significance, NS - Non-significant
1113
impact of the pesticides on the hematological parameters of
the test fish. There was a distinct reduction in percentage of
Haemoglobin (Hb%) also in between treated and control fish,
which was however not significant statistically.
Blood analysis forms a useful tool for determining physiological
and pathological conditions of the fishes. M. cuchia exposed
to sub lethal concentrations of malathion at laboratory
conditions resulted in a significant decrease in RBCs count
leading to anaemia as a result of inhibition of erythropoiesis,
haemosynthesis and increase in the rate of erythrocyte
destruction in haemopeotic organs. Similar reports have been
made by Sampath et al. (1998) on Oreochromis mossambicus
after exposure to zinc and copper. Srivastava (1979) have
shown that exposure to sub lethal concentration of lade
produced haemolytic anaemic condition leading to the lysis
of erythrocytes, Hb content, RBC count and PCV values,
resulting in hypochronic anemia due to deficiency of iron
and decreased utilization for Hb synthesis.
In the present study on M. cuchia exposed to sub lethal
concentration of malathion, significant reduction in RBC count
was recorded causing macrocytic anaemia. While the DLC
revealed decrease in percentage Eosinophil and neutrophils,
there was significant increase in lymphocytes and monocyte
count (Table 2). The increased leucocytes count in treated
fishes may be due to the inclusion of thrombocytes in the
WBC population. Although the difference in mean
haemoglobin percentage is not significant between the treated
and controlled groups, yet the difference reflects the ailing
condition of the test group.
The anaemic condition recorded in the present study could
be due to the destruction of mature RBC by the effect of
pesticide as well as inhibition of erythrocyte production. Such
a decrease in RBC and anaemic response was earlier observed
by Koundinya and Ramamurthy (1979) in Sarotherodan
mossambicus and Tilapia mossambicus after exposure to lethal
concentration of a similar pesticide Sumithion, Lal et al. (1986)
recorded similar findings in H. fossilis exposed to malathion.
In the present study it was apparent that the reduction in the
number of RBC and Hb content had hampered oxidative
metabolism in the test fish. Since the Hb and RBCs are oxygen
carrying devices, the quantitative decrease in their levels might
have lead to the derangement of the oxidative metabolism
with a decrease in cellular activities with respiratory potential.
The weak and lethargic stage of the treated fishes might be the
reason of the reduction in the oxygen level in the body.
The neutrophil increase in fish exposed to malathion may be
due to tissue damage. Mahajan and Dheer (1979) and Sampath
et al. (1998) reported that neutrophils showed great sensitivity
to changes in the environment and increase in the nutriphil
number the important indicators of distressed condition. The
increase in lymphocytes and monocytes suggests that these
fish become stimulated under pesticide stress to fight against
the pollutants in the environment. However, in the differential
count there was a gradual decline in the number of
lymphocytes in fish exposed to malathion for a longer duration
(20 days and 25 days). This significant finding may be due to
continuous stress. The present observation is in accordance
with Mc Leay (1973) who suggested that continuous stress
might lead to a decrease in the number of circulating
lymphocytes.
The results in the present study indicated that prolong exposure
to Malathion has lead to significant increase in the total number
of leucocytes and while the RBC count, Hb concentration
gradually declined as time went by. This phenomenon
suggests that exposure to malathion even at sub lethal dose
cause anaemic and macrocytic changes in M. cuchia which
becomes severe upon prolong exposure.
The findings indicate that malathion 50 EC has harmful effects
on the non-target organisms like M.cuchia at a very low
concentration, leading to serious impact on the hematological
parameters of the infected fish. Although there was no
immediate mortality of the test fishes, such weak health status
would lead to secondary disease infection, low growth, low
metabolism, low reproductive development and will impact
on the overall population status of the species in the natural
resources. Hence, increasing necessity has been realized to
take step for controlling application of chemical pesticides in
agricultural operation and strategy for alternative (biological
or organic) pest control measures may be adopted in paddy
field to safeguard the aquatic biota.
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EFFECT OF MALATHION ON HEMATOLOGICAL PARAMETERS OF MONOPTERUS CUCHIA
1114
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INDIRA  BARUAH  et  al.,


EFFECT OF SUBLETHAL CONCENTRATION OF MALATHION ON HEMATOLOGICAL PARAMETERS OF MONOPTERUS CUCHIA (HAMILTON-BUCHANAN)

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EFFECT OF SUBLETHAL CONCENTRATION OF MALATHION ON HEMATOLOGICAL PARAMETERS OF MONOPTERUS CUCHIA (HAMILTON-BUCHANAN)

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