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In vitro cell-toxicity screening as an alternative animal model for coral toxicology: effects of heat stress, sulfide, rotenone, cyanide, and cuprous oxide on cell viability and mitochondrial function

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Abstract

The logistics involved in obtaining and maintaining large numbers of corals hampers research on the toxicological effects of environmental contaminants for this ecologically and economically important taxon. A method for creating and culturing single-cell suspensions of viable coral cells was developed. Cell segregation/separation was based on specific cell densities and resulting cell cultures were viable for at least 2 mos. Low-density cells lacking symbiotic zooxanthallae and rich in mitochondria were isolated and cultured for toxicity studies. Cells were exposed to differing degrees or concentrations of heat stress, rotenone, cyanide, sulfide, and cuprous oxide. Cells were assayed for mitochondrial membrane potential using the fluorescent probe, JC-9, and for overall viability using the MTT/formazan spectrophotometric viability assay. Significant differences were observed between controls and treatments and the efficacy of this method was validated; only 2 cm2 of tissue was required for a seven-point concentration-exposure series.

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Acknowledgments

We thank Dr. David Julian for instruction in conducting sulfide-exposure experiments and measuring sulfide.

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Authors and Affiliations

  1. Haereticus Environmental Laboratory, P.O. Box 92, Clifford, VA, 24533, USA

    Craig A. Downs & Virgil D. Downs

  2. Office of Public Health Studies, John A. Burns School of Medicine, University of Hawaii—Manoa, 1960 East-West Road, BioMed Tower, Honolulu, HI, 96822, USA

    Craig A. Downs

  3. Department of Biology, University of Central Florida, 4000 Central Florida Boulevard, Orlando, FL, 32816-2368, USA

    John E. Fauth

  4. Pacific Biosciences Research Center, University of Hawaii—Manoa, 2500 Campus Road, Hawaii Hall 211, Honolulu, HI, 96822, USA

    Gary K. Ostrander

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Downs, C.A., Fauth, J.E., Downs, V.D. et al. In vitro cell-toxicity screening as an alternative animal model for coral toxicology: effects of heat stress, sulfide, rotenone, cyanide, and cuprous oxide on cell viability and mitochondrial function. Ecotoxicology 19, 171–184 (2010). https://doi.org/10.1007/s10646-009-0403-5

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