Impact of Curacron Toxicity on Lactate Dehydrogenase in the Serum of the Fish Cyprinus carpio
S. Justin Raj
The experiment was conducted on fish, Cyprinus carpio to study the effect of curacron on serum biomarker (LDH). Curacron is a organophosphate pesticide and is used by the farmers to protect their crops. This pesticide reaches to the aquatic ecosystem by many ways and affects the aquatic fauna. LC50 of Curacron for Cyprinus carpio has been calculated by the log-dose/probit regression line method and recorded as 0.38 ppm at 96 h. Three sub-lethal concentrations (0.1, 0.01 mL and 0.001 ml L-1) were selected to expose the fish for 1, 7, 14 and 21 days. Changes in enzyme activity were observed with all concentrations and exposure period. Lactate concentrations were significantly lower in the experimental group, compared with the control group. The above results of blood plasma profile indicate a marked cytotoxic and hepatotoxic effect of curacron in fishes.
The widespread environmental pollution caused by the chemical substances such
as pesticides is a serious problem for creatures including human (McClure
et al., 2001; McKinlay et al., 2008;
Boobis et al., 2008). The pollutants that enter
the inshore waters and estuaries create serious problems causing extensive damage
to the life and activities of the living aquatic organisms and even to mass
mortality (Joseph et al., 2010). Various chemical
substances entering animal bodies are carried to the organs responsible for
detoxification, such as liver and kidney and excreted. Pesticides are major
cause of concern for aquatic environment because of their toxicity, persistency
and tendency to accumulate in the organisms (Joseph and
Raj, 2010). The impact of these pesticides on aquatic organisms is due to
the movement of pesticides from various diffuse or point sources. These pesticides
are posing a great threat to aquatic fauna especially to fishes, which constitute
one of the major sources of protein rich food for mankind (Sharma
and Singh, 2007). The pesticides induce its effects first at cellular or
even at molecular level, but ultimately it tends to create biochemical disorder
that may even cause death. The organophosphate pesticides modify the activity
of several metabolic enzymes (Radhaiah and Rao, 1990).
Succinic dehydrogenase and Lactate dehydrogenase are the major oxidative enzymes
in carbohydrate metabolism. It may be used for demonstrating tissue damage in
fish. A significant decrease in LDH activity under sub-lethal toxicity of quinolphos
in different tissues of freshwater fish, C. punctatus was reported by
Sastry and Siddiqui (1984). Ganathy
et al. (1994) documented decrease in LDH activity in different tissues
of C. punctatus exposed to hexachlorocyclohexane. Amali
(1995) reported a reduced activity with increasing concentration of quinphos
and paddan in Labeo rohita. The fish serves as bio-indicator of water
quality and the impact of the pesticide can be well understood by analyzing
either blood or serum of the fish, because blood is a pathophysiological reflecter
of whole body (Sharma and Singh, 2004, 2006).
Fishes are widely used to evaluate the health of aquatic ecosystems because
pollutants build up in the food chain and are responsible for adverse effects
and death in the aquatic systems (Farkas et al., 2002;
Joseph et al., 2010). Common carp is widely used
species in aquaculture for food supply in Iran. It provides a good model to
study responses and possible adaptations of local fish populations exposed to
various pollutant sources (Afaghi et al., 2007).
In the present study, the fish, Cyprinus carpio was investigated to evaluate
the effect of Organo phosphorus pesticide Curacron on cytotoxic and hepatotoxic
MATERIALS AND METHODS
Specimens of Cyprinus carpio were obtained from local vicinity in the
year 2007 and introduced into large cement tank (6'X4'X3') disinfected with
potassium permanganate and washed thoroughly prior to introduction of fish (to
prevent fungal infection). Fish were acclimatized for about 20 days before the
commencement of the experiment. They were fed on commercial fish food which
was given daily at morning hours. LC50 of curacron was calculated
by the log-dose/Probit regression line method (Finney, 1971)
and was recorded. The test fishes were grouped in three groups (A, B and C)
having three different sub-lethal concentrations (A = 0.1 mL, B = 0.01 mL and
C = 0.001 ml L-1) for specific time of period (1, 7, 14 and 21 days).
Simultaneously a control set was run to compare the toxicated values. Blood
samples were collected after severing the caudal peduncle and taken in sterilized
centrifuge tubes and allowed to stand for about 1 h, then centrifuged at 2000
rpm for 30 min. The supernatant was separated by a fine rubber bulb pipette
in separate test tube and was used for the estimation of LDH by the method of
RESULTS AND DISCUSSION
In Cyprinus carpio, the serum LDH activity will decrease from the control
set. The LDH activity was found to be decrease with increasing exposure period.
Further, the LDH activity was maximum 25.5" 0.66 when Cyprinus carpio
reared in control. As shown in the Table 1 the LDH activity
decreased from 22.5" 0.66 to 15.97" 0.13, 16,4" 0.49 to 12.5" 0.65 and 19.34"
0.20 to 11.55" 0.33 in 0.1, 0.01 and 0.001 ml L-1. There is a significant
decrease in the percentage variation of LDH with increasing exposure period
due to the toxic effect of curacron. The organophosphate pesticides modify the
activity of several enzymes. It is well known that tissue damaged by toxicants
exhibit a sharp rise in activity of mitochondrial enzymes aspartate transferase
and alanine amino transferase (Abdelsalam et al.,
1982; Mikhail et al., 1979).
the concentration of enzyme Lactate dehydrogenase of control and curacron
treated Cyprinus carpio in different exposure periods. The results
are expressed as F moles of formazon formed mg dL-1 protein
|Percentage in parentheses. Values are in means±SE
Alanine amino transferase and aspartate amino transferase activities of the
blood increase significantly due to curacron poisoning. Alanine amino transferase
has been strongly implicated in the production of energy in tissues and is considered
as a stress indicator (Gould et al., 1976). Asparate
amino transferase is the main transaminase that interfere with TCA cycle in
a major way Lowenstein (1967).
A rise in its activity indicates the occurrence of greater energy demand, which
are normally associated with synthetic activities of the cell (Meister,
1955). In this study, the LDH activity in the serum of Cyprinus carpio
decreased when exposed to different concentrations of curacron. The inhibition
of LDH activity indicated may be due to the functioning of in intermediates
in to the TCA cycle. This might be responsible for the suppression of the oxidative
phase of tissue metabolism under curacron impact. The intoxication of these
pesticides combine with an enzyme to form an enzyme inhibition complex which
react with various functional groups of the enzymes inhibit the normal enzyme
activity of major metabolic site. Similar inhibited LDH activity under various
pesticide stresses are reported in fishes by Sastry and
Siddiqui (1984) and Kabeer et al. (1983).
However, Natarajan et al. (1994) has reported
stimulation in LDH activity in response to metasytox and chlordane will be studied
in tissues of Puntias conchonius.
The study depicts that the pesticide curacron is highly toxic to the fish Cyprinus carpio and the stress responses showed by fish are dependent on concentration and duration of exposure. LDH is as a general indicator of the existence and severity of acute or chronic tissue damage and sometimes, as a monitor of progressive conditions like hemolytic anaemia From the present study, it may be concluded that the analysis of enzyme activity of fishes can effectively be used as an indicator of fish health. Long term exposure of organisms to pesticides means a continuous health hazard for the population. So, human population is at high risk by consuming these toxicated fishes. It is also suggested that these type of toxicological studies are required to monitor the aquatic life and predict the toxic effect of pesticides on aquatic organisms particularly fish.
1: Abdelsalam, E.B., S.E.I. Adam and G. Tartour, 1982. Mixture of dieldrin and phosphamidon rises GOT, GPT and suppress cholinesterase in blood serum. Zentralale Veternaemed Reihe. A., 29: 136-138.
2: Afaghi, A., S. Zare, R. Heidari, Y. Asadpoor and R.M. Viayeh, 2007. Effects of copper sulfate (CuSo4) on the levels of glucose and cortisol in common carp, Cyprinus carpio. Pak. J. Biol. Sci., 10: 1655-1660.
CrossRef | PubMed | Direct Link |
3: Amali, A.A., 1995. Biochemical and enzymological studies on the toxicity of pesticides (Quinolphos and padan) in the fresh water fish, Labeo rohita (HAM.). Ph.D. Thesis, University of Madras. Tamil Nadu, India.
4: Boobis, A.R., B.C. Ossendorp, U. Banasiak, P. Hamey, Y. Sebestian and A. Moretto, 2008. Cumulative risk assesment of pesticide residues in food. Toxicol. Lett., 180: 137-150.
5: Farkas, A., J. Salanki and A. Specziar, 2002. Relation between growth and the heavy metal concentration in organs of bream, Abramis brama L. populating lake Balaton. Arch. Environ. Contam. Toxicol., 43: 236-243.
CrossRef | PubMed | Direct Link |
6: Finney, D.J., 1971. Probit Analysis. 3rd Edn., Cambridge University Press, London, UK., pp: 76-80
CrossRef | Direct Link |
7: Ganathy, V.D., S. Reddy, S.L.N. Reddy and K. Shankariah, 1994. Effect of hexachlorocyclohexane. J. Ecotoxical, Environ. Monit., 4: 15-20.
8: Gould, E., R.S. Collier, J.J. Karoulus and S.A. Givenus, 1976. Heart transaminase in the rock crab, cancer irrortus exposed to cadmium salts. Bull. Environ. Contam. Toxicol., 15: 635-643.
9: Joseph, B. and S.J. Raj, 2010. Effect of curacron toxicity on the total serum protein content of Cyprinus carpio. Toxicol. Environ. Chem., 92: 1889-1893.
CrossRef | Direct Link |
10: Joseph, B., S.J. Raj, B.T. Edwin, P. Sankarganesh, M.V. Jeevitha, S.U. Ajisha and S.R. Sheeja, 2010. Toxic effect of heavy metals on aquatic environment. Int. J. Biol. Chem. Sci., 4: 939-952.
CrossRef | Direct Link |
11: Kabeer, A.I.K., R.S. Sambasiva and K.V.R.R. Rao, 1983. Dehydrogenase systems under augmented sub-lethal Malathion stress. J. Anim. Morphol. Physiol., 30: 101-106.
12: Lowenstein, J.M., 1967. Metabolic Pathway. Vol. I, Academic Press, New York, pp: 147
13: McClure, G.Y., R.M. Helm, K. Stine, A.W. Burks, S.M. Jones and J. Gandy, 2001. Evaluation of immune parameters in propanil-exposed farm families. Arch. Environ. Contam. Toxicol., 41: 104-111.
14: McKinlay, R., J.A. Plant, J.N. Bell and N. Voulvoulis, 2008. Calculating human exposure to endocrine disrupting pesticides via agricultural and non-agriculturl exposure routes. Sci. Total Environ., 398: 1-12.
15: Meister, A., 1955. Methods Enzymology. Vol. 2, Academic Press, New York, pp: 283
16: Mikhail, T.H., N. Aggour, K. Awadallah, M.N. Boules, E.A. Eldessoukey and A.J. Karim, 1979. Dursban resulted rise of GOT, GPT and simultaneous decrease of cholinesterase. Z. Ernachrung Swiss, 18: 258-262.
17: Natarajan, V., H.N. Jayaram, W.M. Scribner and J.G. Garcia, 1994. Activation of endothelial cell phospholipase D by sphingosine and sphingosine-1-phosphate. Am. J. Respir. Cell Mol. Biol., 11: 221-229.
18: Radhaiah, V. and K.J. Rao, 1990. Toxicity of phsethroid insecticide fenvalerate to a freshwater fish. Tilapia mossambica, Changes in glycogen metabolism of muscle. Ecotox. Environ. Safety, 19: 116-116.
19: Sastry, K.V. and A.A. Siddiqui, 1984. Some haematological, biochemical and enzymological parameters of freshwater fish, Channa punctatus exposed to Sub-lethal concentration of quinolphos. Pestic. Biochem. Physiol., 22: 8-13.
20: Sharma, G. and S. Singh, 2004. Studies on the effect of intoxicant indofil on the blood morphology of Channa punctatus (Bloch.). Bionotes, 6: 20-20.
21: Sharma, G. and S. Singh, 2006. Assay of some blood parameters of the fish, Channa punctatus (Bloch.) after intoxication of Indofil. Bionotes, 8: 21-21.
22: Sharma, G. and S. Singh, 2007. Effect of indofil toxicity on MCHC of Channa punctatus (Bloch.). J. Environ. Res. Dev., 1: 261-263.
Direct Link |
23: Tietz, W., 1976. Ascorbic acid. In: Fundamentals of Clinical Chemistry. Tietz, W. (Ed.). W.B. Saunders Company, Britain, pp: 422-419