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20211
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Subject: Veterinary Pharmacology and Toxicology × Author: DABHOLKAR AKSHAY RAJKUMAR × End date: 2023 × Start date: 2020 ×
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    ThesisItemOpen Access
    CYTOTOXIC, CARCINOGENIC AND GENOTOXIC EFFECTS OF CARBOSULFAN IN CULTURED MAMMALIAN CELLS
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, THRISSUR, 2021-12-30) DABHOLKAR AKSHAY RAJKUMAR; Bibu John Kariyil
    Carbosulfan a carbamate class of insecticide, is commonly used in agricultural practices for soil, foliar and seed treatment. There is a paucity of data about the cytotoxic, carcinogenic and genotoxic effects of carbosulfan. The data so obtained could be useful in elucidating the various health hazards in humans and animals due to the environmental exposure to carbosulfan. Hence the present study was envisaged to evaluate the cytotoxic, carcinogenic and genotoxic effects of carbosulfan in cultured mammalian cells. The cytotoxic effect was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay in CHO-K1 cells. The mean cell viability per cent of carbosulfan at 12.5, 25, 50, 100, 250 and 500 µg/mL was found to be 96.93 ± 2.40, 95.66 ± 3.36, 91.89 ± 5.31, 73.28 ± 3.08, 34.24 ± 1.67 which proved carbosulfan to be cytotoxic. Carbosulfan treatment at 100, 250 and 500 µg/mL showed loss of basic cytoskeletal structure and cellular contact with reduction in cell density, cell shrinkage, cell detachment and debris formation. Carbosulfan treatment showed induction of oxidative stress as revealed by significant (p ≤ 0.001) increase in the intracellular reactive oxygen species (ROS) generation in CHO-K1 cells. The DNA fragmentation studies showed significant (p ≤ 0.001) concentration dependent increase in the per cent of DNA fragments in carbosulfan treated CHO K1 cells. It was observed to produce significant (p ≤ 0.001) DNA damage in comet assay as evident by significant (p ≤ 0.001) increase in tail length, tail moment and olive tail moment. Carbosulfan treatment at 50 µg/mL showed time dependent carcinogenicity in BALB/c 3T3 clone A31 cells using cell transformation assay revealing the carcinogenic potential of carbosulfan. Genotoxicity measured using in vitro chromosomal aberration test revealed that carbosulfan treatment induced significant (p ≤ 0.001) concentration and time dependent chromosomal aberrations like breaks, gaps and exchange in chromatid, breaks, gaps and exchange in chromosome, chromosome fragments, dicentric chromosome, centromeric disruption, ring formation, pulverised chromosome and multiple aberrations. In vitro micronucleus formation test also showed significant (p ≤ 0.001) concentration and time dependent increase in the number of binucleate cells with micronuclei (BNMN). The above results proved carbosulfan to be genotoxic. Hence the present study concluded that carbosulfan produced cytotoxicity, carcinogenicity and genotoxicity in cultured mammalian cells due to the ROS mediated oxidative stress.