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In the present paper we determined low molecular heavy metal binding protein called metallothionein (MT) in fish spermatozoa sampled from four fish species (Acipenser baerii, Acipenser ruthenus, Perca fluviatilis and Sander lucioperca). The spermatozoa samples were analyzed with differential pulse voltammetry DPV (Brdicka reaction). The average content of MT in the fish sperm was 0.64 +- 0.55 micro m. The highest content of MT was determined in spermatozoa of S. lucioperca (0.96 +- 0.86 micro m), followed by P. fluviatilis (0.88 +- 0.11 micro m), A. ruthenus (0.57 +- 0.60 micro m) and A. baerii (0.43 +- 0.25 micro m). The differences in the content of MT was probably related to the differences in age and perhaps to the health status based on the environmental conditions in which the tested specimens developed while also other still unknown factors may have been involved.

Summary Glutathione belongs to a vital intra- and extra-cellular protective antioxidant and is found almost exclusively in its reduced form. The ratio between its reduced and oxidized within cells is often used as a marker of cellular toxicity. The objectives of the study were to (i) determine both the reduced (GSH) and oxidized glutathione (GSSG) and cysteine (Cys) in the sperm of the Acipenser baerii and Acipenser ruthenus ,a s well as in Perca fluviatilis and Sander lucioperca, and (ii) to demonstrate the differences in concentration levels between representatives of acipenseriform and teleost species. High performance liquid chromatography with electrochemical detection was employed. The average content of the thiols determined in the sperm samples were as follows: Acipenser baerii (cysteine 55 ± 8 l gm l )1 ; GSH 126 ± 19 l gm l )1 ; GSSG 49 ± 7 l gm l )1 ), Acipenser ruthenus (cysteine 62 ± 9 l gm l )1 ; GSH 768 ± 115 l gm l )1 ; GSSG 180 ± 16 l gm l )1 ), Sander lucioperca (cysteine 251 ± 38 l gm l )1 ; GSH 185 ± 28 l gm l )1 ; GSSG 93 ± 14 l gm l )1 ), Perca fluviatilis (cysteine 281 ± 42 l gm l )1 ; GSH 496 ± 74 l gm l )1 ; GSSG 138 ± 21 l gm l )1 ). Based on the results obtained it can be concluded that this method is sensitive and selective for the determination of these compounds in real samples. Results revealed differences in cysteine content between species of the two systematic categories but also showed that ratios between GSH and GSSG can vary between species while potentially predict oxidative stress in fish sperm.