Reduction and methylation of inorganic mercury in Porcellio scaber (Isopoda, Crustacea) and its environment were studied, using a purpose-built experimental setup where Hg cycling was followed using 203Hg2+ tracer in experiments without and with isopods. In experiment without isopods, daily reduction of 203Hg2+ to 203Hg0 under sterile and nonsterile conditions was measured for three weeks to assess the contribution of bacteria to this process. In experiments with isopods, daily release of 203Hg0 was measured for two weeks. Total mercury (T203Hg) and monomethylmercury (Me203Hg) in whole animals, gut, digestive glands (hepatopancreas), food (hazelnut leaves), and feces were measured to obtain the assimilation and distribution of mercury in the animals, to investigate the origin and fate of Me203Hg, and, finally, to assess the mass balance of mercury in the experimental system. Experiment without isopods showed the important role of bacteria in reduction of 203Hg2+ to 203Hg0, especially in the first day of the experiment. Experiments with isopods showed that formation of 203Hg0 depended on the 203Hg2+ concentration in the food. The contribution of the isopod's digestive flora in reduction of 203Hg2+ to 203Hg0 was negligible. Approximately 3% of T203Hg and 2% of Me203Hg consumed was assimilated by the animals. Methylation of 203Hg2+ occurred already in the leaves before they were consumed by the isopods. Assimilation of Me203Hg from the food surprisingly was low. Also, a loss of Me203Hg was noticed when comparing assimilated and excreted Me203Hg versus consumed Me203Hg. This may be explained by the assumption that demethylation of MeHg prevailed over methylation of Hg2+ in the animal's digestive system, leading to excretion of ingested mercury as Hg2+.
Two established methods for assessment of the cytotoxicity of contaminants, the lysosomal latency (LL) assay and the neutral red retention (NRR) assay, were successfully applied to in toto digestive gland tubes (hepatopancreas) of the terrestrial isopod Porcellio scaber (Isopoda, Crustacea). In vitro exposure of isolated gland tubes to copper was used as a positive control to determine the performance of the two methods. Lysosomal latency and the NRR assay were then used on in vivo (via food) laboratory-exposed animals and on field populations. Arbitrarily selected criteria for determination of the fitness of P. scaber were set on the basis of lysosomal membrane stability (LMS) as assessed with in toto digestive gland tubes. Decreased LMS was detected in animals from all polluted sites, but cytotoxicity data were not in agreement with concentrations of pollutants. Lysosomal membrane stability in the digestive gland tubes of animals from an environment in Idrija, Slovenia that was highly polluted with mercury (260 microg/g dry wt food and 1,600 microg/g dry wt soil) was less affected than LMS in laboratory animals fed with 5 and 50 microg Hg/g dry weight for 3 d. This probably indicates tolerance of P. scaber to mercury in the mercury-polluted environment and/or lower bioavailability of environmental mercury. In animals from the vicinity of a thermal power plant with environmental mercury concentrations three to four orders of magnitude lower than those in Idrija, LMS was severely affected. In general, the LL assay was more sensitive than the NRR assay. The LMS assay conducted on digestive gland tubes of terrestrial isopods is highly recommended for integrated biomarker studies.
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