Assessment of genotoxicity through DNA damage analysis in Indian major carps induced by acute exposure to Chakbandi drain water

Paper Details

Research Paper 01/03/2018
Views (284) Download (10)

Assessment of genotoxicity through DNA damage analysis in Indian major carps induced by acute exposure to Chakbandi drain water

Muhammad Arshad Rana, Tayyaba Sultana, Salma Sultana, Bilal Hussain
J. Bio. Env. Sci.12( 3), 169-180, March 2018.
Certificate: JBES 2018 [Generate Certificate]


In this study, Chakbandi drain’s composite water concentrations were collected from the selected sites in the month of April, May and June, 2016 and applied to fingerling’s of three Indian major carps i.e. Catla catla, Labeo rohita and Cirrhinus mrigala under laboratory conditions in glass aquaria. After determining the LC50, the sub-lethal dilutions i.e. 20%, 25%, 30%, 35% and 40% of drain water were tested for three month’s acute toxicity trial. All the water quality parameters i.e. DO, pH, temperature, conductivity, TDS, salinity, TSS, BOD and COD and concentration of selected heavy metals (Cu, Cr, Mn, Cd, Co, Ni, Sn, Hg, Zn and Pb) recorded from drain water were found above the permissible limits as described by FAO/ WHO. During current research, the DNA damage in fish peripheral erythrocytes was quantified by using different categories of damaged cells that were arbitrarily defined according to the tail length (size) of the comets. Maximum frequency of DNA damaged cells was recorded in erythrocytes of Cirrhinus mrigala when compared with Labeo rohita and Catla catla. Cirrhinus mrigala showed maximum average tail DNA length and average tail moment than Labeo rohita and Catla catla. Conclusively, DNA fragmentation as biomarker approach was found to be reliable for the assessment of genotoxicity and environmental pollution. Moreover, findings of this study are helpful as an early warning for environmental monitoring strategies.


Adeyemo OK. 2005. Haematological and histopathological effects of Cassava Mill Effluent in Clarias gariepinu. African Journal of Biomedical Research 8, 179 -183.

Al-Sabti K, Metcalfe CD. 1995. Fish micronuclei for assessing genotoxicity in water. Mutation Research 343(2), 121-35.

Andrade VM, Silva J, Silva FR, Heuser VD, Dias JF, Yoneama ML, Freitas TR. 2004. Fish as bioindicators to assess the effects of pollution in two southern Brazilian rivers using the Comet assay and Micronucleus test. Environmental and Molecular mutagenesis 44(5), 459-68.

Atienzar AF, Cordi B, Donkin ME, Evenden AJ, Jha AN, Depledge MH. 2000. Comparison of ultraviolet-induced genotoxicity detected by random amplified polymorphic DNA with chlorophyll fluorescence and growth in a marine macroalgae, Palmaria palmate. Aquatic Toxicology 50(2), 1-12.

Bistodeau TJ, Barber LB, Bartell SE, Cediel RA, Grove KJ. 2006. Larval exposure to environmentally relevant mixtures of al-kylphenole thoxylates reduces reproductive competence in male fathead minnows. Aquatic  Toxicolology  79(3), 268-77.

Bolognes C, Cirillo S. 2014. Genotoxicity biomarkers in aquatic bio indicators. Current Zoology 60(2), 273–284.

Bolognesi C, Landini E, Roggieri P, Fabbri R, Viarengo A. 1999. Genotoxicity biomarkers in the assessment of heavy metal effects in mussels: experimental studies. Environmental Molecular Mutagenesis 33(4), 287-292.

Bonga WSE. 1997. The stress response in fish. Physiology Revolution 77, 591-625.

Carrasco K, Tilbury KL, Myers MS. 1990. Assessment of the piscine micronucleus test as an in situ biological indicator of chemical contaminant effects. Canadian journal of Fisheries and Aquatic Sciences 47(11), 2123-2136.

Cavas T, Konen S. 2007. Detection of cytogenetic and DNA damage in peripheral erythrocytes of goldfish (Carrassius auratus) exposed to glyphosate formulation using the micronucleus test and comet assay. Mutagenesis 22, 263-272.

Devaraj S, Arulprakasam C, Kandhan AP, Neelamegam K, Kalaiselvan R. 2014. Toxicological effects of ammonia on gills of Cyprinus carpio var. communis (Linn.). Journal of Coastal Life Medicine 2(2), 94-98.

Dhawan A, Bajpayee M, Pandey AK, Parmar D. 2009. Protocol for the single cell gel electrophoresis/comet assay for rapid genotoxicity assessment. Industrial Toxicology Research Centre Marg, Lucknow, India pp. 153-159.

Dikilitas M, Kocyigit A, Yigit A. 2009. Molecular-based fast method to determine the extent of DNA damages in higher plants and fungi. African Journal of Biotechnology 8(14), 3118-3127.

El-Bassir AHA, Ashjgan MA, Maab EE, Ryan SG, Alim DIM. 2017. Levels of four heavy metals in two fish species collected from Aldubaseen area in khartoum state, sudan. EC Nutrition 6(4), 130-134.

Evans DH, Piermanin PM, Choe KP. 2005. The Multifunctional fish gill: dominant site of gas exchange, osmoregulation, acid base regulation and excretion of nitrogenous waste. Physiology Revolution 85, 97-177.

Gebel TW. 2001. Review: Genotoxicity of arsenical compounds. International Journal of Hygenic Environmental Health 203, 249-262.

Giesy JP, Pierens SL, Snyder EM, Miles-Richardson S, Kramer VJ. 2000. Effects of 4-nonylphenol on fecundity and biomarkers of estrogenicity in fathead minnows Pimephales promelas. Environmental Toxicology 19, 1368-1377.

Ginebreda A, Kuzmanovic M, Guasch H, López de Alda M, López-Doval JC et al. 2014. Assessment of multi-chemical pollution in aquatic ecosystems using toxic units: Com-pound prioritization, mixture characterization and relationships with biological descriptors. Science Total Environment 469, 715-723.

Goksoyr A, Anderson T, Buhler DR, Stegeman JJ, Williams DB, Forlin L. 1991. Immunochemical cross reactivity of (3-naphthoflavone inducible cytochrome P450 (P450 IAI) in lives microsomes from different fish species and rat. Fish Physiology and Chemistry 9(1), 1-13.

Greig SM, Sear DA, Carling PA. 2005. The impact of fine sediment accumulation on the survival of incubating salmon progeny: implications for sediment management. Science and Total Environment 344, 241-258.

Grisolia CK, Rivero CLG, Starling FLRM, da-Silva ICR, Barbosa AC, Dorea JG. 2009. Profile of micronucleus frequencies and DNA damage in different species of fish in a eutrophic tropical lake. Genetics and Molecular Biology 32(1), 138- 143.

Gupta V, Malik DS, Dinesh K. 2017. Risk assessment of heavy water pollution in middle stretch of river Ganga: an introspection. International Research Journal of Environment Science 6(2), 62-71.

Hayat S, Javed M, Razzaq S. 2007. Growth performance of metal stressed major carps viz. Catla catla, Labeo rohita and Cirrhinus  mrigala reared under semi-intensive culture system. Pakistan Veterinary Journal 27(1), 8-12.

Javed M. 2005. Growth responses of Catla catla, Labeo rohita and Cirrhina mrigala for bio-accumulation of zinc during chronic exposure. Pakistan Journal of Biological Science 8, 1357- 1360.

Kousar S, Javed M. 2015. Diagnosis of metals induced DNA damage in fish using comet assay. Pakistan Veterinary Journal 35(2), 168-172.

Kumar PS, Ananthan G, Kumar DS, Jagadeesan L. 2011. Haematology and biochemical parameters of different feeding behavior of teleost fishes from vellar Estuary, India. Comparative Clinical Pathology 11, 1257-1259.

Lee HB, Peart TE. 2000. Bisphenol A contamination in Canadian municipal and industrial wastewater and sludge samples. Water Quality Research Journal of Canada 35, 283–298.

Lopes PA, Pinheiro T, Santos MC, da Luz Mathias M, Collares-Pereira MJ, Viegas-Crespo AM. 2001. Response of antioxidant enzymes in freshwater fish populations (Leuciscus alburnoides) to inorganic pollutants exposure. Science and Total Environment 280, 153-163.

Mallin MA, McIver MR, Fulton M, Wirth E. 2011. Elevated levels of metals and organic pollutants in fish and clams in the cape fear river watershed. Archives of Environmental Contamination and Toxicology 61, 461 – 471.

Mohamad EA, Osman AR. 2014. Heavy metals concentration in water, muscle and gills of Oreochromis niloticus collected from the sewage treated water and the White Nile. International Journal of Aquaculture 4(6), 36-42.

Moustafa MZ, Al-Sayed EM. 2014. Impact of water pollution with heavy metals on fish health: Overview and updates. Global Veterinaria 12(2), 219-231.

Naz S, Javed M. 2013. Growth responses of fish during chronic exposure of metal mixtures under laboratory conditions. Pakistan Veterinary Journal 33, 354-357.

Nhapi I, Wali UG, Uwonkunda BK, Nsengimana H, Banadda N, Kimwaga BK. 2011. Assessment of water pollution levels in the Nyabugogo Catchment, Rwanda. The Open Environmental Engineering Journal 4, 40-53.

Noor N, Zutshi B. 2016. Bioaccumulation of trace metals in tissues of rohu fish for environmental risk assessment. Journal of Water Resource and Protection 8, 472-481.

Nussey G, Vuren JHJ, Preez HH. 1995. Effect of copper on the differential white blood cell counts of the Mozambique tilapia (Oreochromis mossambicus). Comparative Biochemistry and Physiology 111, 381-388.

Obiakor MO, Okonkwo JC, Nnabude PC, Ezeonyejiaku CD. 2012. Eco-genotoxicology: Micronucleus assay in fish erythrocytes as in situ aquatic pollution biomarker: a review. Journal of Animal Sciences 2, 123-133.

Perera PACT, Suranga P, Kodithuwakku TV, Sundarabarathy & Edirisinghe U. 2015.  Bioaccumulation of cadmium in freshwater fish: an environmental perspective. Insight Ecology 4(1), 1-12, 2015.

Ramesh F, Agarajan K. 2013. Evaluation of DNA damage of the fish Clarias batrachus exposed to Sago effluent using the comet assay. European International Journal of Science and Technology 2(3), 12-16.

Ruane NM, Bonga WSE, Balm PHM. 1999. Differences between rainbow trout and brown trout in the regulation of the pituitary inter-renal axis and physiological performance during confinement. Comparative Endocrinology 115, 210-219.

Sabullah MK, Ahmad SA, Shukor MY, Gansau AJ, Syed MA, Sulaiman MR, Shamaan NA. 2015. Heavy metal biomarker: Fish behavior, cellular alteration, enzymatic reaction and proteomics approaches. International Food Research Journal 22(2), 435-454.

Santamaría WC, Gómez MS, Santamaría YMV. 2016. Using genotoxic and haematological biomarkers as an evidence of environmental contamination in the Ocoa River native fish, Villavicencio meta, Colombia. Springer Plus 5, 351-356.

Shakir HA, Qazi JI, Chaudhry AS. 2015. Assessing human  health  risk  of  metal  accumulations  in  a  wild  carp  fish  from selected  sites  of  a  river  loaded  with  municipal  and  industrial  wastes. International Journal of Environmental Research 9(2), 545-552.

Shirani M, Mirvaghefi A, Farahm H, Abdollahi M. 2012. Biomarker responses in mudskipper (Periophthalmus waltoni) from the coastal areas of the Persian Gulf with oil pollution. Environmental Toxicology and Pharmacology 34(3), 705-713.

Singh NP, McCoy MT, Tice RR, Schneider EL. 1988. A simple technique for quantification of low levels of DNA damage in individual cells. Experimental Cellular Research 175(1), 184-191.

Sultana T, Butt K, Sultana S,   Al-Ghanim KA, Mubashra R, Bashir N, Ahmed Z, Ashraf A, Mahboob S. 2016a. Histopathological changes in liver, gills and intestine of Labeo rohita inhabiting industrial waste contaminated water of river Ravi. Pakistan Journal of Zoology 48(4), 1171-1177.

Villela IV, Oliveira IM, Silva J, Henriques JAP. 2006. DNA damage and repair in hemolymph cells of golden mussel (Limnoperna fortunex) exposed to environmental contaminants. Mutation Research 605(2), 78-86.

Vosyliene MZ, Jankaite A. 2006. Effect of heavy metal model mixture on rainbow trout biological parameters. Ekologija Niger 4, 12-17.

Wilson RW, Taylor EW. 1993. The physiological responses of freshwater rainbow trout, Onchorynchus mykiss, during acute exposure. Journal of Comparative Physiology 163(2), 38-47.

Wood CM, Pessy SF, Wright PA, Bergman HL, Randall DJ. 1989. Ammonia and urea dynamics in the lake Magadi tilapia a teleost fish adapted to an extremely alkaline environment. Respire Physiology 77, 1 -20.

Yancheva V, Velcheva I, Stoyanova S, Georgieva E. 2016.  Histological biomarkers in fish as a tool in ecological risk assessment and monitoring programs: A review. Applied Ecology and Environmental Research 14(1), 47-75.

Zaniboni–Filho E, Noner APO, Reynalte–Ta- taye DA, Serafini RL. 2009. Water pH survival. Fish Physiology and Biochemistry 35, 151-155.

Zaqoot HA, Aish AM, Wafi HN. 2017. Baseline concentration of heavy metals in fish collected from Gaza fishing harbor in the Mediterranean sea along Gaza Coast, Palestine. Turkish Journal of Fisheries and Aquatic Sciences 17, 101-109.

Zizy I, Elbialy Ismail T, Doaa H, Abdelhady Amel M, Elasely. 2016. Assessment of genotoxic effects of pesticide residues and related haemato-biochemical parameters on farmed Nile Tilapia (Oreochromis niloticus L.) in Kafrelsheikh Governorate, Egypt. Alexandria Journal of Veterinary Sciences 44(1), 136-146.