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Submitochondrial particles asin vitro biosensors of heavy metal toxicity


The effects on mitochondrial respiratory parameters of heavy metals, such as Cu, Ni, Pb, Cd, Zn, Ag, Hg, were recorded by using thein vitro response of submitochondrial particles (SMP) from beef heart mitochondria.

The toxicity of these elements was estimated by determining their effects on the energy-coupled reverse electron transfer (RET), which is induced by ATP and succinate at first site level of the respiratory chain in SMP.

The RET rate was easily monitored by recording spectrophotometrically at 340 nm the production of NADH, arising from the reduction of exogenous NAD+ by RET.

The toxicity values were expressed as the toxicant molar concentration which decreases the rate of reduction of NAD+ to an extent of 50 percent (EC50). The toxicity increased in the following order: Ni2+<Pb2+<Zn2+< Cd2+<Hg2+<Cu2+<Ag+.

The SMP data were compared with the toxicity values obtained from a variety of biological systems currently used for toxicity testing. The results obtained demonstrate that the SMP test generally provides a good estimate of metal toxicity for several fish and invertebrate species. This is demonstrated by the statistical parameters obtained in the regression analysis. The broadened 95% confidence intervals and, in particular, the poor correlations obtained for some aquatic organisms can be ascribed to the more complex metabolic interactions and competing toxic pathways in aquatic organisms, when compared to SMP.

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  • Argese, E., A Marcomini, P. Miana, C. Bettiol & G. Perin, 1994. Submitochondrial particle response to linear alkylbenzene sulfonates, nonylphenol polyethoxylates and their biodegradation derivatives. Env. Toxicol. Chem. 13(5): 737–742.

    Google Scholar 

  • Argese, E., C. Bettiol, A. Ghelli, R. Todeschini & P. Miana, 1995. Submitochondrial particles as toxicity biosensors of chlorophenols. Env. Toxicol. Chem. 14(3): 363–368.

    Google Scholar 

  • Blondin, G. A., L. M. Knobeloch, H. W. Read & J. M. Harkin, 1987. Mammalian mitochondria asin vitro monitors of water quality. Bull. Environ. Contam. Toxicol. 38: 467–474.

    Google Scholar 

  • Denizeau, F., M. Marion, M. Chtaib & J. P. Schmit, 1990. Toxicity of heavy metals in cultured hepatocytes. Environ. Toxicol. Chem. 9: 737–743.

    Google Scholar 

  • Dutka, B. J. & K. K. Kwan, 1981. Comparison of three microbial toxicity screening tests with the Microtox test. Bull. Environ. Contam. Toxicol. 27: 753–757.

    Google Scholar 

  • Forstner, U. & G. T. W. Wittman, 1981.Metal Pollution in the Aquatic Environment. Springer-Verlag. New York, pp. 1–26.

    Google Scholar 

  • Hansen, L. & L. Smith, 1964. Studies on mechanism of oxidative phosphorylation. VII. Preparation of a submitochondrial particle (ETPH) which is capable of fully coupled oxidative phosphorylation. Biochim. Biophys. Acta 81: 214–222.

    Google Scholar 

  • Joshi, S. & J. B. Hughes, 1981. Inhibition of coupling factor B activity by cadmium ion, arsenite-2,3-dimercaptopropanol, and phenylarsine oxide, and preferential reactivation by dithiols. J. Biol. Chem. 256(21): 11112–11116.

    Google Scholar 

  • Keller, A. E. & S. G. Zam, 1991. The acute toxicity of selected metals to the freshwater mussel,Anodonta imbecilis. Environ. Toxicol. Chem. 10: 539–546.

    Google Scholar 

  • Martin, T. R. & D. M. Holdich, 1986. The acute lethal toxicity of heavy metals to peracarid crustaceans (with particular reference to fresh-water asellids and gammarids). Wat. Res. 20(9): 1137–1147.

    Google Scholar 

  • Muller, L., 1986. Consequences of cadmium toxicity in rat hepatocytes: mitochondrial dysfunction and lipid peroxidation. Toxicology 40: 286–295.

    Google Scholar 

  • Nacci, D., E. Jackim & R. Walsh, 1986. Comparative evaluation of three rapid marine toxicity tests: sea urchin early embryo growth test, sea urchin sperm cell toxicity test and Microtox. Environ. Toxicol. Chem. 5: 521–525.

    Google Scholar 

  • Piekering, Q., D. O. Carle, A. Pill, T. Willingham & J. Lazorchak, 1989. Effects of pollution on freshwater organisms. J. Wat. Poll. Contr. Fed. 61(6): 998–1042.

    Google Scholar 

  • Pilli, A., D. O. Carle, E. Kline, Q. Pickering & J. Lazorchak, 1988. Effects of pollution on fresh water organisms. J. Wat. Poll. Contr. Fed. 60(6): 994–1065.

    Google Scholar 

  • Schubauer-Berigan, M. K., J. R. Dierkes, P. D. Monson & G. T. Ankley, 1993. pH-dependent toxicity of Cd, Cu, Ni Pb and Zn toCerodaphnia dubia, Pimephales promelas, Hyalella atzeca and Lumbricus variegatus. Environ. Toxicol. Chem. 12: 1261–1266.

    Google Scholar 

  • Vallee, B. L. & D. D. Ulmer, 1972. Biochemical effects of mercury, cadmium, and lead. Annu. Rev. Biochem. 41: 91–128.

    Google Scholar 

  • Yagi, T. & Y. Hatefi, 1984. Thiols in oxidative phosphorylation: inhibition and energy-potentiated uncoupling by monothiol and dithiol modifiers. Biochemistry 23: 2449–2455.

    Google Scholar 

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Argese, E., Bettiol, C., Miana, P. et al. Submitochondrial particles asin vitro biosensors of heavy metal toxicity. Journal of Aquatic Ecosystem Health 5, 125–134 (1996).

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Key words

  • mitochondria
  • reverse electron transfer
  • aquatic toxicology
  • screening test
  • metal toxicity assessment