The chromosomal aberration test in kidney cells of Channa punctatus was conducted to study the genotoxic effect of synthetic pyrethroid fenvalerate, Different chromosomal abnormalities were seen and their frequency was recoded in metaphase spreads obtained from fishes exposed to three sub lethal concentrations (0.0625, 0.0314 and 0.0157 ppm) of this compound. The exposure to fenvalerate caused various structural abnormalities in chromosomes such as chromatid break, fragment, gap, chromatid separation, deletion and ring type chromosomes. Results obtained in present investigations clearly indicate that fenvalerate acts as a genotoxic agent in Channa punctatus

Chaudhari, Renu

Saxena, Kamal Kumar

Journal of Applied and Natural Science

70Study of chromosomal abnormalities in Channa punctatus exposed to fenvalerateKamal Kumar Saxena* and Renu ChaudhariPest and Parasite Research Laboratory, Department of Zoology, Bareilly College, Bareilly-243005(U.P.), INDIA*Corresponding author. E-mail: drkamal06@yahoo.co.inAbstract: The chromosomal aberration test in kidney cells of Channa punctatus was conducted to study thegenotoxic effect of synthetic pyrethroid fenvalerate, Different chromosomal abnormalities were seen and theirfrequency was recoded in metaphase spreads obtained from fishes exposed to three sub lethal concentrations(0.0625, 0.0314 and 0.0157 ppm) of this compound. The exposure to fenvalerate caused various structuralabnormalities in chromosomes such as chromatid break, fragment, gap, chromatid separation, deletion and ringtype chromosomes. Results obtained in present investigations clearly indicate that fenvalerate acts as a genotoxicagent in Channa punctatus.Keywords:  Fenvalerate, Chromosomal aberration, Genotoxic effect, Channa punctatusINTRODUCTIONPesticides are being used very commonly throughoutthe world for various purposes. These are main harmfulpollutants among the chemicals released by man into theenvironment. In an ideal situation, a pesticide should fallexactly on the target and get degraded completely intoharmless compounds.  As per the estimate of Piementaland Leviton (1986) only less than 1% of the pesticidesused reaches target and the remaining part spreads inthe surroundings which in turn affects the non targetorganisms and cause pollution in the environment. Fishprovides a suitable model for monitoring aquaticgenotoxicity because of their ability to metabolizexenobiotics and accumulate pollutants (Grisolia andCordeiro, 2000). Fishes can also respond to mutagens atlow concentration of toxicants in a manner similar to highervertebrates (Al-Sabti and Metcalfe, 1995).The changes caused by genotoxins in the genetic materialcan be detected in fish at specific level by using variousgenotoxic assay. In fish kidney chromosomes thechromosome aberration test (CAT) is most suitable tostudy the genotoxic effect of pesticides. Syntheticpyrethroids are reported (Saxena and Seth, 2002; Sethand Saxena, 2003, Sirohi and Saxena, 2006 and Saxenaand Sirohi, 2007) to be highly toxic to fish because ofthese compounds are strongly absorbed by the gills evenat very low concentration in water due to their highlipophilicity. These compounds possess moderatemammalian toxicity but they are extremely toxic to fishand aquatic invertebrates (Eisler, 1992).The study of genotoxic effect of pesticides and otherpollutants in fishes is very important. Some workers (Rishiand Grewal, 1995; Arockia-Rita and Selvanayagam, 1998;Chauhan et al., 2000; Devi et al., 2001; Aboul, 2002; Poliet al., 2003; Lioi et al., 2004; Saxena and Rana, 2005 andAli et al., 2008) have reported genotoxic effects of somepesticides and other pollutants on mammals and fishes.In present study an attempt has been made to study theeffect of fenvalerate on chromosomes of Channapunctatus.MATERIALS AND METHODSThe common fresh water fish Channa punctatus werecollected from the fresh water resources of Bareilly andadjoining places and acclimatized to laboratoryconditions for few days in untreated soft water. Afteracclimation the fish were divided in to experimental andcontrol groups. The experimental group fishes wereexposed to different concentrations of fenvalerate (0.0625,0.0314 and 0.0157 ppm).Chromosomal preparations were made according to themethod of Ojima (1982), Nagpure and Barat (1997) andAsano et al. (1998). After exposure to fenvalerate, theinjection of 0.05% Colchicine (1ml per 100 gm body wt.)was given to the fishes and they were left for 2 hrs. Afterthis time duration the kidneys were excised from thetreated fishes and were immediately transferred toseparate Petri dishes containing freshly prepared 0.56%KCl (hypotonic solution). After processing the tissuesthe suspension was prepared and centrifuged.After discarding the supernatant, remaining part wasfixed with methanol acetic acid fixative. These slides wereprepared by dropping method and after staining withGiemsa stain cells were covered with D.P.X. Thechromosomes were observed by using 100x oil immersionmicroscope and their photographs were taken.ISSN : 0974-9411 All Rights Reserved © Applied and Natural Science Foundation   www.ansfoundation.orgJournal of Applied and Natural Science 2 (1): 70-73 (2010)  JANS2008APPLIED    ANDNATURAL SCIENCEFOUNDATIONANSF71RESULTS AND DISCUSSIONIn the present investigations changes in kidney cells ofChanna punctatus have been observed due to exposureto sub lethal concentrations of fenvalerate. The changein chromosomal structures directly affects the geneticmaterial of the animals and by examining the metaphasestructure of chromosomes after exposure to pesticideslike fenvalerate, the clastogenic properties of the pesticidecan be detected (Ali et al., 2009). In Channa punctatus32 chromosomes were observed in normal diploid cellsin both sexes. No sex chromosomes were observed inthis fish species. These observations are in agreementwith the earlier data available (Rishi and Haobam, 1990and Rishi and Grewal, 1995, Saxena and Rana, 2008). Inpresent study prominent changes in the structure ofchromosomes, such as decrease in length, increase inbreadth and over all compression in size was observed.Some important chromosomal aberrations were chromatidbreaks, fragments, gaps, chromatid separation, acentricfragments, deletion and ring type chromosomes.The data obtained indicate that the frequencies ofchromosomal abnormalities were higher during the initialperiod of exposure and there after declined. However,they were still higher than the frequencies in controlsamples (Manna and Mukherjee, 1986). It was alsorecorded that as the exposure period increased, thepercentage of chromosomal abnormalities also increased.When the fishes were exposed for 96 hours, themetaphase spreads had only two types of abnormalities.Chromosomal breaks were observed more than any othertype chromosomal abnormalities when fish Channapunctatus were exposed to 144 hrs. (Mathew andJahageerdar, 1999) have also reported similar observationsin Channa punctatus exposed to heavy metals.Kamal Kumar Saxena and Renu Chaudhari / J. Appl. & Nat. Sci. 2 (1): 70-73 (2010)Concentration of fenvalerate (in ppm) Exposure period  (in hours) No. of fishes dissected  No. of metaphase spreads studied No. of metaphase spreads with abnormalities % of metaphase spreads with abnormalities 0.0000 (Control) 0 24 48 72 96 120 144 5 5 5 5 5 5 5 50 50 50 50 50 50 50 0 0 1 2 1 2 2 0 0 2 4 2 4 4 0.0157 24 48 72 96 120 144 5 5 5 5 5 5 50 50 50 50 50 50 1 7 10 16 13 6 2 14 20 32 26 12 0.0314 24 48 72 96 120 144 5 5 5 5 5 5 50 50 50 50 50 50 3 10 19 23 17 11 6 20 38 46 34 22 0.0625 24 48 72 96 120 144 5 5 5 5 5 5 50 50 50 50 50 50 6 11 29 37 21 19 12 22 58 74 42 38 Table 1.  Frequency of chromosomal aberrations induced by fenvalerate in kidney cells of Channa punctatus.72Acute and sublethal toxicity are the important parametersfor the evaluation of pesticidal activity. Mortality dependsupon the concentration and duration of exposure. Dataobtained in present studies indicate that fenvalerate ishighly toxic to Channa punctatus, as it caused mortalityat the 0.10 ppm. In the present studies, LC50 offenvalerate for Channa punctatus was also determinedsince the study of sublethal toxicity of pesticides is moreimportant than the acute toxicity (Alderic, 1967) becauseit gives an idea about the mode of action of the compoundinside the body of fish.During fenvalerate treatment, Channa punctatus showedseveral changes in their behaviour. This behaviourresponse varied in accordance with the concentration offenvalerate. At higher concentrations they exhibitederratic swimming. They often lost their balance anddashed against the wall of aquaria and even tried toescape out. The fishes often came to the surface of waterto gulp atmospheric air, which may be to avoid the toxicenvironment. Opercular activity was increased afterexposure to fenvalerate. In the present investigationsthe colour of fish became pale and this change was alsodependent on the concentration of fenvalerate. Thesefindings are in agreement with Seth and Saxena (2003).The change in chromosomal structures directly affectsthe genetic material of the animals. The DNA repair isreported (Matsumura, 1985) to be slower in fishes thanmammals therefore, these changes appear to be useful indetecting genotoxicity caused by aquatic pollutants infishes. By examining the metaphase structure ofchromosomes after exposure to pesticides likefenvalerate, the clastogenic properties of the pesticidecan be detected. It was observed that chromosomes inexposed fishes were reduced in length in comparison tothe chromosomes of control fishes. Aquatic organismsare susceptible to the chemicals present in theirsurroundings and if these chemicals cause DNA damage,it may result into a variety of abnormalities like reducedgrowth, abnormal development and age of adult andlarval stages. Chromosomal mutations may causecarcinogenesis and the development of varioussyndromes (Bond and Rossbacher, 1993).The percentage of metaphase spreads with abnormalitieswas also very high i.e. more than 70% at all theconcentrations used. The abnormalities were significantat the level of Pd”0.01. More than three types ofaberrations were also observed in a single metaphasespread in several cases. The abnormalities dependedupon both exposure period and dose level of fenvalerate.The results presented in this study clearly show thatexposure to fenvalerate, even at very low concentrationscause chromosomal abnormalities, which are directlyassociated with DNA damage. It shows a high level ofgenotoxic potential of fenvalerate for this fish species.This may affect over all fish health and in turn may alsoaffect other animals including human beings which areconsuming the fish as food.REFERENCESAboul, Ela. E.I. (2002). Genotoxic effect of rifampicin on themouse. Bull. Mat. Res. 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Toxic effect of ë-cyhalothrinon biochemical contents of fresh water fish Channapunctatus. J. Fisheries Aquatic Sci., 1(2): 112-116.Kamal Kumar Saxena and Renu Chaudhari / J. Appl. & Nat. Sci. 2 (1): 70-73 (2010)

Study of chromosomal abnormalities in Channa punctatus exposed to fenvalerate

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Study of chromosomal abnormalities in Channa punctatus exposed to fenvalerate

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