title: In vitro cytotoxicity assessment of two heavy metal salts in a fish cell line (RF).
abstract: Labeo rohita (rohu) is one of most important of Indian major carps, which is highly ranked among the important freshwater aquaculture species in the world. Heavy metals are major risk factors for aquatic health, which are biomagnified through the food chain. The present study was carried out to investigate the effect of different divalent salts of two heavy metals, such as zinc (ZnCl(2), ZnSO(4) and ZnNO(3)) and cadmium (CdCl(2) and CdSO(4)), in an established fish cell line, RF developed from fin tissue of L. rohita. The RF cell line was used for assessment of heavy metal cytotoxicity through various endpoint assays, including maximum tolerated dose (MTD) determination, 3-[4,5-dimethylthiazole-2-yl]-2,5-diphenyl tetrazolium bromide (MTT) assay, neutral red (NR) uptake assay, and Coomassie brilliant blue (CBB) assay. Results revealed that these heavy metal salts were cytotoxic to the RF cell line at varied concentrations. MTD values were found to be 1.563, 3.125, 6.25, 12.5 and 25 mg/L for CdCl(2), CdSO(4), ZnCl(2), Zn(NO(3))(2) and ZnSO(4), respectively. The half-maximal inhibitory concentration values calculated by MTT, NR and CB assay were 53.83 ± 7.02, 58.03 ± 9.12 and 79.20 ± 15.27 for ZnSO(4), 26.44 ± 7.01, 36.60 ± 7.82 and 155.6 ± 14.75 for Zn(NO(3))(2), 20.26 ± 17.95, 16.94 ± 7.05 and 87.54 ± 7.58 for ZnCl(2), 5.166 ± 0.57, 15 ± 1 and 41.80 ± 8.38 for CdSO(4) and 4.966 ± 0.56, 9.56 ± 1.73 and 20.93 ± 4.47 for CdCl(2). This study establishes the RF cell line as an in vitro tool for assessment and monitoring of heavy metal concentration in the aquatic environment.
authors.affiliation: National Bureau of Fish Genetic Resources , Lucknow , India .
authors.forename: Mukunda
authors.initials: M
authors.lastname: Goswami
authors.forename: Kamalendra
authors.initials: K
authors.lastname: Yadav
authors.forename: Akhilesh
authors.initials: A
authors.lastname: Dubey
authors.forename: Bhagwati S
authors.initials: BS
authors.lastname: Sharma
authors.forename: Rituraj
authors.initials: R
authors.lastname: Konwar
authors.forename: Ravindra
authors.initials: R
authors.lastname: Kumar
authors.forename: Naresh S
authors.initials: NS
authors.lastname: Nagpure
authors.forename: Wazir S
authors.initials: WS
authors.lastname: Lakra
mesh_terms: D000818:Animals; D002104:Cadmium / Q000633:toxicity*; D002347:Carps / Q000378:metabolism*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004784:Environmental Monitoring / Q000379:methods*; D020714:Maximum Tolerated Dose; D009499:Neutral Red; D012394:Rosaniline Dyes; D013778:Tetrazolium Salts; D013844:Thiazoles; D018675:Toxicity Tests; D015032:Zinc / Q000633:toxicity*
chemical_list: D012394:Rosaniline Dyes; D013778:Tetrazolium Salts; D013844:Thiazoles; D002104:Cadmium; D009499:Neutral Red; C048139:Coomassie blue; C022616:thiazolyl blue; D015032:Zinc