Nuclear Abnormalities in Erythrocytes of Frogs from Wetlands and Croplands of Western Ghats Indicate Environmental Contaminations

Authors

  • Shreyas Raghunath Kuvempu University
  • Chethankumar Masaruru Veerabhadrappa
  • Sannanegunda Venkatarama Bhatta Krishnamurthy Kuvempu University

DOI:

https://doi.org/10.11594/jtls.07.03.04

Keywords:

Agro-chemicals, Euphlyctis cyanophlyctis, micronucleus, nuclear abnormalities, Western Ghats-India

Abstract

Anuran amphibians are the biological models to assess the influence of environmental contamination. We conducted nuclear abnormality assessment and micronuclei test in erythrocytes of frogs to identify an early influence of environmental contaminations. In Western Ghats of India, farmers use different agrochemicals and obviously, the amphibian habitat is contaminated with combinations of many residues. Many frog species use these agro-ecosystem for breeding and to complete early life stage. In the present study, we used Indian skipper frog (Euphlyctis cyanophlyctis (Anura: Ranidae)), a common inhabitant of water bodies in agro-ecosystems of Western Ghats for the assay. We collected the adult frogs from three different habitats; a) the rice paddy fields contaminated with agro-chemicals, b) uncontaminated marsh and c) a shallow water pool. We recorded micronucleus, blebbed, lobed, kidney bean shaped nucleus and nucleus with notches in erythro-cytes of these frogs. These nuclear abnormalities in erythrocytes are quantified.  The incidences of occurrence of micronucleus ranged from 0 to 8 in 1000 analyzed cells.  More than 80% of the frogs living in agro-chemical contaminated site showed these abnormalities. The appearance of nuclear abnormalities indicates that the frogs are affected by agro-chemicals.

References

Mann RM, Hyle RV, Choung CB, Wilson SP (2009) Am-phibians and agricultural chemicals: Review of the risks in a complex environment. Environmental Pollution 157 (11): 2903 – 2927. doi: 10.1016/j.envpol.2009.05.015.

Ismail M, Khan QM, Ali R et al. (2014) Evaluation of the genotoxicity of chlorpyrifos in common indus valley toad, Bufo stomaticus using alkaline single-cell gel electrophore-sis (comet) assay. Agricultural Science 5: 376 – 382. doi: 10.4236/as.2014.54039.

Lips KR (1998) Decline of a tropical montane amphibian fauna. Conservation Biology 12 (1): 106 – 112.

Venturino A, Rosenbaum E, Decastro AC et al. (2003) Biomarkers of effects in toads and frogs. Biomarker 8 (3 – 4): 167 – 186. doi: 10.1080/1354700031000120116.

Falfushinska HI, Romanchuk LD, Stolyar OB (2008) Dif-ferent responses of biochemical markers in Frogs (Rana ridibunda) from urban and rural wetlands to the effect of carbamate fungicide. Comparative Biochemistry and Phys-iology Part C: Toxicology and Pharmacology 48 (3): 223 – 229. doi: 10.1016/j.cbpc.2008.05.018.

Sewell D, Griffiths RA (2009) Can a single Amphibian species be a good biodiversity indicator? Diversity 1 (2): 102 – 117. doi:10.3390/d1020102.

Leiva-Presa A, Mortensen AS, Arukwe A, Jenssen BM (2006) Altered hepatic Retinol and CYP26 levels in adult European common frogs (Rana temporaria) exposed to p, p’- DDE. Marine Environmental Research 62 (Supplement 1): 510 – 515. doi: 10.1016/j.marenvres.2006.04.013.

Krishnamurthy SV (2003) Amphibian assemblages in un-dis-

turbed and disturbed areas of Kudremukh National Park, Central Western Ghats, India. Environmental Conserva-tion

(3): 274 – 282. doi: 10.1017/S0376892903000274.

Naniwadekar R, Vasudevan K (2007) Patterns in diversity of anurans along an elevational gradient in the Western Ghats, South India. Journal of Biogeography 34 (5): 842 – 853. doi: 10.1111/j.1365-2699.2006.01648.x.

Vasanthi K, Chairman K, Singh AJAR, Raj AJK (2014) Amphibian diversity and distribution in Courtallam, South Western Ghats foothills, India. International Journal of Biodiversity Conservation 6 (4): 351 – 362. doi: 10.5897/IJBC2013.0675.

Gurushankara HP, Krishnamurthy SV, Vasudev V (2007) Morphological abnormalities in natural populations of common frogs inhabiting agro-ecosystems of central West-ern Ghats. Applied Herpetology 4: 39 – 45. doi: 10.1163/157075407779766651.

Krishnamurthy SV, Meenakumari D, Gurushankara HP, Vasudev V (2008) Nitrate induced morphological anoma-lies in the tadpoles of Nyctibatrachus major and Fejervarya limnocharis (Anura: Ranidae). Turkish Journal of Zoology 32: 239 – 244.

Patel AM, Kulkarni PA, Girish KG et al. (2008) Fejervarya limnocharis Morphology. Herpetological Review 31 (9): 77.

Vasudev V, Krishnamurthy SV, Gurushankara HP (2008) Organophosphate pesticides-a major threat to anuran pop-ulations in an agro ecosystems of Western Ghats, India. Froglog 83: 8-9

Hegde G, Krishnamurthy SV (2014) Analysis of health status of the frog Fejervarya limnocharis (Anura: Ranidae) living in rice paddy fields of Western Ghats, using body condition factors and AchE content. Ecotoxicology and Environmental Contamination 9 (1): 69-76. doi: 10.5132/eec.2014.01.009.

Gurushankara HP (2007) Cytogenetical and biochemical effects of organophosphate pesticides malathion and para-thion on Rana (Limnonectus) Limnocharis. Ph.D Thesis. Kuvempu University.

Muranli FDG, Guner U (2011) Induction of micronuclei and nuclear abnormalities in erythrocytes of mosquito fish (Gambusia affinis) following exposure to the pyrethroid in-secticide λ -cyhalothrin. Mutation Research/Genetic Toxi-cology and Environmental Mutagenesis 726 (2): 104 – 108. doi: 10.1016/j.mrgentox.2011.05.004.

Muranli FDG (2013) Genotoxic and cytotoxic evaluation of pyrethroid insecticides λ-cyhalothrin and α-cypermethrin on human blood lymphocyte culture. Bulletin of Environ-mental Contamination and Toxicology 90: 357 – 363. doi: 10.1007/s00128-012-0909-z.

Venturino A, Pechen De Angelo AM (2005) Biochemical targets of xenobiotics: Biomarkers in amphibian ecotoxi-cology. Applied Herpetology 2: 335 – 353. doi: 10.1163/1570754054507433.

Josende ME, Tozetti AM, Alalan MT et al. (2015) Geno-toxic evaluation in two amphibian species from Brazilian subtropical wetlands. Ecological Indicators 49: 83 – 87.

Mouchet F, Gauthier L, Mailhes C et al. (2006) Compara-tive evaluation of genotoxicity of Captan in amphibian lar-vae (Xenopus laevis and pleurodeles waltl) using the com-et assay and the micronucleus test. Environmental Toxi-cology 21 (3): 264 – 277. doi: 10.1002/tox.20180.

Ferrier V, Gauthier L, Zoll-Morreux CL, Haridon J (1998) Genotoxicity testing in amphibians: a review. In: Mi-croscale testing in aquatic toxicology: Advance techniques and Practice. Philadelphia, CRC Press, pp 507 – 519.

Da Rocha CAM (2011) The micronucleus test in erythro-cytes of amphibian larvae as tool for xenobiotic exposure site assessment: A brief review and an example using Lithobates catesbeianus exposed to copper sulphate. Mid-dle-East Journal of Scientific Research 8 (1): 23 – 29.

Amphibia Web (2017) Amphibian. http://amphibiaweb.org/. Accessed: July 2017.

Hegde G (2014) Use of agrochemicals and their influence on population structure of Anuran amphibians in agro-ecosystems of Western Ghats. Ph.D Thesis. Kuvempu Uni-versity.

Lajmanovich RC, Cabagna M, Peltzer PM et al. (2005) Micronucleus induction in erythrocytes of the Hyla pul-chela tadpoles (Amphibia: Hylidae) exposed to the insecti-cide endosulfan. Mutation Research/Genetic Toxicology and Environmental Mutagenesis 587 (1 – 2): 67 – 72. doi: 10.1016/j.mrgentox.2005.08.001.

Carrasco KR, Tilbury TL, Myers MS (1990) Assessment of piscine micronuclei test as an in situ biological indicator of chemical contaminant effects. Canadian Journal of Fisher-ies and Aquatic Sciences 47 (11): 2123 – 2136. doi: 10.1139/f90-237.

Strunjak-Perovic IR, Coz-Rakovac, Topicpopovic N, Jadan M (2009) Seasonality of nuclear abnormalities in gilthead sea bream Sparus aurata (L) erythrocytes. Fish Physiology and Biochemistry 35 (2): 287 – 291. doi: 10.1007/s10695-008-9208-3.

Burlibasa L, Gavrila L (2011) Amphibians as model organ-isms for study environmental genotoxicity. Applied Ecolo-gy and Environmental Research 9 (1): 11-15. doi: 10.15666/aeer/0901_001015.

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Published

2017-09-11

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