An Investigation on Genotoxic Effects of Arsenic on Caspian Roach (Rutilus Caspicus) Using Micronucleus and Comet Assays

Subject Areas : Environmental Toxicology

Mahsa Daneshian ¹ , Hadiseh Kashiri ² , ali akbar hedayati ³ , Kaveh Khosraviani ⁴

1 - M.S.c of Aquatic Ecology, Faculty of Fisheries and Environment, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
2 - Assistant Professor, Faculty of Fisheries and Environment, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.* (Corresponding Author)
3 - Associate Professor, Faculty of Fisheries and Environment, Gorgan University of Agricultural Sciences and Natural Resources, Gorgan, Iran.
4 - M.S.c of Fisheries. Natural Resources Faculty of Karaj, Tehran University, Teharn, Iran.

Received: 2017-12-26 Accepted : 2018-05-09 Published : 2021-04-21

Keywords: Micronucleus, Arsenic, Genotoxic, Comet, Rutilus caspicus,

Abstract :

Background and Objective: Arsenic, as a pollutant, can lead to serious problems in aquatic organisms and human. In the present study, considering the economic and ecological value of Caspian roach, Rutilus caspicus, genotoxic effects of arsenic on this species were assessed using the comet and micronucleus assays in 2016.Method: At first, the fish were exposed to the concentrations of 3, 6 and 9 mg L-1 arsenic. A control treatment without arsenic was also considered. Sampling was done during the 3 and 7th exposure days. To perform micronucleus test, blood sampling was done at two times and micronuclei frequency was determined. To perform comet assay, sampling was done from the fish gill tissue and the DNA damage parameters were assessed.  Findings: Exposure to the arsenic led to the induction of micronuclei formation in the erythrocytes and DNA damage in the gill tissue. So that micronuclei frequency and values of damage parameters in the treatments exposed to arsenic significantly increased compared to the control (P≤0.05). The induced damage level was dependent on the exposure time and concentration. The highest levels of damage parameters including Tail Length, Tail Moment, %Tail DNA and also micronuclei frequency were 101.1±1.55, 57.11±1.64, 59.45±3.09 and 2.26±0.58, respectively which observed in the treatment exposed to 9 mg L-1 arsenic.Discussion and Conclusion: The results from the present study indicated sub-lethal concentrations of arsenic can cause micronuclei formation and induction of DNA damage in Caspian roach. Therefore, it could be stated that arsenic has genotoxic effects on Caspian roach. 

References:

1. Kiabi, B.H., Abdoli, A., Naderi, M., 1999. Status of the fish fauna in the South Caspian basin of Iran. Journal of Zoology in the Middle East, Vol. 18, pp. 57-65.


2. De­Mora, S., Sheikholeslami, M.R., Wyse, E., Azemard, S., Cassi, R., 2004. An assessment of metal contamination in coastal sediments of the Caspian Sea. Marine Pollution Bulletin, Vol. 48, pp. 61-77.


3. Di Gioacchino, M., Petrarca, C., Perrone, A., Farina, M., Sabbioni, E., Hartung, T., Martino, S., Esposito, D.L., Lotti, L.V. Mariani-Costantini, R., 2008. Autophagy as an ultrastructural marker of heavy metal toxicity in human cord blood hematopoietic stem cells. Science of the Total Environment, Vol. 392, pp. 50-58.


4. Bagheri, H., Darvish Bastami, K., Sharmad, T., Bagheri, Z., 2013. Assessment of the distribution of heavy metals pollution in Gorgan bay. Oceanography, Vol. 3(11), pp. 65-72. (In Persian)


5. Hashemi, S.J., Riahi Bakhtiari, A.R., Lak, R., 2013. Source identification and distribution of lead, copper, zinc, nickel, chromium and vanadium in surface sediments of Caspian Sea. Journal of Mazandaran University of Medical Sciences, Vol. 23(1), pp. 36-50. (In Persian)


6. Bagheri, H., Azimi, A., 2016. Study of the heavy metals distribution in surface sediments of Sisangan coasts-south of Caspian Sea. Oceanography, Vol. 6(21), pp. 27-36. (In Persian)


7. Saghali, M., Baqraf, R., Patimar, R., Hosseini, S.A., Baniemam, M., 2014. Determination of heavy metal (Cr, Zn, Cd and Pb) concentrations in water, sediment and benthos of the Gorgan Bay (Golestan province, Iran). Iranian Journal of Fisheries Sciences, Vol. 13(2), pp. 449- 455.

1. Ventura-Lima, J., Bogo, M.R., Monserrat, J.M., 2011. Arsenic toxicity in mammals and aquatic animals: A comparative biochemical approach. Ecotoxicology and Environmental Safety, Vol. 74, pp. 211-218.


2. Zhang, W., Huang, L., Wang, W.X., 2011. Arsenic bioaccumulation in a marine juvenile fish Terapon jarbua. Aquatic Toxicology, Vol. 105, pp. 582-588.


3. Chakraborti, D., 2011. Arsenic: Occurrence in Groundwater. In: Nriagu JO (eds), Encyclopedia of Environmental Health. Burlington, Elsevier. pp. 165-80.


4. Kumari, B., Kumar, V., Sinha, A.K., Ahsan, J., Ghosh, A.K., Wang, H., DeBoeck, G., 2016. Toxicology of arsenic in fish and aquatic systems. Environmental Chemistry Letters, Vol. 15(1), pp. 43-64.


5. Bagheri, H., 2017. Study of arsenic distribution in sediments of the southeastern Caspian Sea. Journal of Marine Sciences and Technology, Vol. 79, pp. 35-42. (In Persian)  

13. World Health Organization, 2001. Environmental Health Criteria, arsenic and arsenic compounds. 114p.

1. Vanitha, S., Amsath, A., Muthukumaravel, K., Sugumaran, J., 2017. Effect of Arsenic on hematological parameters of freshwater fish, Channa punctatus (Bloch). International Journal of Zoology and Applied Biosciences, Vol. 2(3), pp. 117-121.


2. Kumar, R., Banerjee, T.K., 2016. Arsenic induced hematological and biochemical responses in nutritionally important cat fish Clarias batrachus (L.). Toxicology Reports, Vol. 3, pp. 148-152.


3. Hallauer, J., Geng, X., Yang, H.C., Shen, J., Tsai, K.J., Liu, Z., 2016. The effect of chronic Arsenic exposure in zebrafish. Zebrafish, Vol. 13(5), pp. 405-412.


4.  Li, C., Li, P., Tan, Y.M., Lam, S.H., Chan, E.C.Y., Gong, Z., 2016. Metabolic characterizations of liver injury caused by acute Arsenic toxicity in zebrafish. PLOS ONE, Vol. 11(3), e0151225.

18. Siu, W.H.L., Caob, J., Jack, R.W., Wu, R.S.S., Richardson, B.J., Xu,, Lam, P.K.S., 2004. Application of the comet and micronucleus assays to the detection of B[a]P genotoxicity in haemocytes of the green-lipped mussel (Perna viridis). Aquatic Toxicology, Vol. 66, pp. 381-392.


19. Gravato, C., Santos, M.A., 2003. Genotoxicity biomarkers’ association with B(a)P biotransformation in Dicentrarchus labrax Ecotoxicology and Environmental Safety, Vol. 55, pp. 352-358.

1. Bolognesi, C., Perrone, E., Roggieri, P., Sciutto, A., 2006. Bioindicators in monitoring long term genotoxic impact of oil spill: Haven case study. Marine Environmental Research, Vol. 62, pp. 287-291.

21. Mizuno, K., Shinohara, N., Miyakoshi, J., 2015. In Vitro Evaluation of genotoxic effects under magnetic resonant coupling wireless power transfer. International Journal of Environmental Research and Public Health, Vol. 12, pp. 3853-3863.

1. Ramirez, O.A.B., Garcia, F.P., 2005. Genotoxic damage in zebra fish (Danio rerio) by arsenic in waters from Zimapan, Hidalgo, Mexico. Mutagenesis, Vol. 20(4), pp. 291-295.


2. Ahmed, M.K., Al-Mamun, H., Hossain, M.A., Arif, M., Parvin, E., Akter, M.S., Khan, M.S., Islam, M., 2011. Assessing the genotoxic potentials of arsenic in tilapia (Oreochromis mossambicus) using alkalin comet assay and micronucleus test. Chemosphere, Vol. 84, pp. 143-149.


3. Das, S., Unni, B., Bhattacharjee, M., Wann, S.B., Rao, P.G., 2012. Toxicological effects of arsenic in a freshwater teleost fish, Channa punctatus. African Journal of Biotechnology, Vol. 11(19), pp. 4447-4454.


4. Kumar, A., Kesari, V.P., Khan, P.K., 2013. Fish micronucleus assay to assess genotoxic potential of arsenic at its guideline exposure in aquatic environment. Biometals, Vol. 26, pp. 337-346.


5.  Sayed, A.E.D.H., Elbaghdady, H.A.M., Zahran, E., 2015. Arsenic-induced genotoxicity in Nile tilapia (Oreochromis niloticus); the role of Spirulina platensis extract. Environmental Monitoring and Assessment, Vol. 187, pp. 1-10.


6. Kousar, S., Javed, M., 2014. Assessment of DNA damage in peripheral blood erythrocytes of fish exposed to arsenic under laboratory conditions. International Journal of Current Microbiology and Applied Sciences, Vol. 3(11), pp. 877-888.


7. Kousar, S., Javed, M., 2016. Determination of Arsenic induced nuclear abnormalities in peripheral blood erythrocytes of carps using micronucleus test. The Journal of Animal and Plant Sciences, Vol. 26(5), pp. 1501-1506.

29. Di Giulio, R.T. Hinton, D.E., 2008. The Toxicology of Fishes. Taylor and Francis. pp. 319–884.


30. Hedayati, S.A.A., Jahanbakhshi, A.R., Qadermarzi, A., Bagheri, T., 2014. Aquatic toxicology. Gorgan University of Agricultural Sciences and Natural Resources, 212 p.


31. Oliveira, M., Pacheco, M., Santos, M.A., 2007. Cytochrome P4501A, genotoxic and stress responses in golden grey mullet (Liza aurata) following short-term exposure to phenanthrene. Chemosphere, Vol. 66, pp. 1284–91.


32. Ayllon, F., Garcia-Vazquez, E., 2000. Induction of micronuclei and other nuclear abnormalities in European minnow Phoxinus phoxinus and mollie Poecilia latipinna: an assessment of the fish micronucleus test. , 467(2), pp. 177-86.


33. Singh, N.P., McCoy, M.T., Tice, R.R., Schneider, E.L., 1988. A simple technique for quantitation of low levels of DNA damage in individual cells. Experimental Cell Research, Vol. 175, pp. 184-191.


34. Tice, R.R., Agurell, E., Anderson, D., Burlinson, B., Hartmann, A., Kobayashi, H., Miyamae Y., Rojas, E., Ryu, J.C., Sasaki , Y.F., 2000. Single cell gel/comet assay: guidelines for in vitro and in vivo genetic toxicology testing. Environmental and Molecular Mutagenesis, Vol. 35(3), pp. 206-221.


35. Konca, K.., Lankoff, A., Banasik, A., Lisowska, H., Kuszewski, T., Gozdz, S., Koza, Z., Wojcik, A.A., 2003. cross-platform public domain PC image-analysis program for the comet assay. Mutation Research, Vol. 534, pp. 15–20.

1. Ullah, S., Hasan, Z., Zorriehzahra, M.J., Ahmad, S., 2017. Diagnosis of endosulfan induced DNA damage in rohu (Labeo rohita, Hamilton) using comet assay. Iranian Journal of Fisheries Sciences, Vol. 16(1), pp. 138-149.


2. Kumar, A., Kesari, V.P., Alok, A.K., Kazim, S.N., Khan, P.K., 2014. Assessment of arsenic-induced DNA damage in goldfish by a polymerase chain reaction-based technique using random amplified polymorphic DNA markers. Archives of Environmental Contamination and Toxicology, Vol. 67(4), pp. 630-638.


3. Selvaraj, V., Armistead, M.Y., Cohenford, M., Murray, E., 2013. Arsenic trioxide (As₂O₃) induces apoptosis and necrosis mediated cell death through mitochondrial membrane potential damage and elevated production of reactive oxygen species in PLHC-1 fish cell line. Chemosphere, Vol. 90, pp. 1201-1209.


4. Sunaina, A., Bhardwaj, A.K., Ansari, B.A., 2016. Oxidative stress biomarkers in assessing arsenic trioxide toxicity in the zebrafish, Danio rerio. International Journal of Fisheries and Aquatic Studies, Vol. 4(4), pp. 08-13. 


5. Asophian, H.V., Asophian, M.M., 2006. Arsenic Toxicology: Five questions. Chemical Research in Toxicology, Vol. 19, pp. 1-15.


6. Engstrom, J., Hedenstierna, G., Larsson, A., 2010. Pharyngeal oxygen administration increases the time to serious desaturation at incubation in acute lung injury: an experimental study. Critical Care, Vol. 14, R93.


7. Ding, W., Liu, W., Cooper, K.L., Qin, X.J., Bergo, P.L.S., Hudson, L.G., Liu, K.J., 2009. Inhibition of poly (ADP-ribose) polymerase-1 by arsenite interferes with repaire of oxidative DNA damage. Journal of Biological Chemistry, Vol. 284(11), pp. 6809-6817.


8. Flora, S.J., Mittal, M., Pachauri, V., Dwivedi, N., 2012. A possible mechanisms for combined arsenic and fluoride induced cellular and DNA damage in mice. Metallomics, Vol. 4, pp. 78-90.


9. Shen, S., Li, X.F., Cullen, W.R., Weinfeld, M., Le, X.C., 2013. Arsenic binding to proteins. Chemical Reviews, Vol. 113, pp. 7769-7792.


10. Ventura-Lima, J., Fattorini, D., Regoli, F., Monserrat, J.M., 2009. Effects of different inorganic arsenic species in Cyprinus carpio (Cyprinidae) tissues after short-time exposure: bioaccumulation, biotransformation and biological responses. Environmental Pollution, Vol. 157, pp. 3479-3484.


11. Parveen, N., Shadab, G.G.H.A., 2012. Cytogenetic evaluation of cadmium chloride on Channa punctatus. Journal of Environmental Biology, Vol. 33, pp. 663-666.

47. Grisolia, C.K., Rivero, C.L.G., Starling, F.L.R. M.,Silva, I.C.R., Barbosa, A.C.B., Dorea, J.G., 2009. Profile of micronucleus frequencies and DNA damage in different species of fish in a eutrophic tropical lake. Genetics and Molecular Biology, Vol. 32(1), pp. 138-14.


48. Rocha, C., Cavalcanti, B., Pessoa, C.O., Cunha, L., Pinheiro, R.H., Bahia, M., Ribeiro, H., Cestari, M., Burbano, R., 2011. Comet assay and micronucleus test in circulating erythrocytes of Aequidens tetramerus exposed to methylmercury. In vivo, Vol. 25(6), pp. 929-933.


49. Bolognesi, C., Perrone, E., Roggieri, P., Sciutto, A., 2006. Bioindicators in monitoring long term genotoxic impact of oil spill: Haven case study. Marine Environmental Research, Vol. 62, pp. 287-291.


50. Yadav, K.K., Trividi, S.P., 2009. Sublethal exposure of heavy metals induces micronuclei in fish, Channa punctatus. Chemosphere, Vol. 77, pp. 1495-1500.


51. Vanzella, T.P., Martinez, C.B.R., Colus, I.M.S., 2007. Genotoxic and mutagenic effects of diesel oil water fraction on a neotropical fish species. Mutation Research, Vol. 631, pp. 36–43.


52. Udroiu, I., 2006. The micronucleus test in piscine erythrocytes. Aquatic Toxicology, Vol. 79, pp. 201-204.


53. Al-Sabti, K.., Metcalfe, C.D., 1995. Fish micronuclei for assessing genotoxicity in water. Mutation Research, Vol. 343(23), pp. 121-135.