Heat shock protein expression patterns (HSP70) in the hydrothermal vent mussel Bathymodiolus azoricus

We previously reported evidence of increased levels of DNA damage in the hydrothermal mussel Bathymodiolus azoricus, which suggested that the species was not fully resistant to the natural toxicity of its deep-sea vent environment. In the present study, HSP70 was used as a biomarker of sub-cellular stress. Differences in HSP70 expression pattern were observed between vent sites, typified by different depths/toxicity profiles, and between different mussel tissue types. A comparison of specimens collected by remote operated vehicle (ROV) and acoustically-operated cages showed that less stress (as indicated by changes in HSP70 levels) was induced by the faster cage recovery method. Therefore alternatives to ROV collection should be considered when planning experiments involving live deep sea organisms. Significantly, a positive correlation was found between the levels of DNA strand breakage, as measured using the Comet assay, and HSP70 expression pattern; evidence was also obtained for the constitutive expression of at least one HSP isoform which was located within the cell nucleus

The effects of hydrostatic pressure change on DNA integrity in the hydrothermal-vent mussel Bathymodiolus azoricus: implications for future deep-sea mutagenicity studies

Comet and agarose gel electrophoresis (AGE) assays were used to show that haemocytes (blood cells) and gill tissues of vent mussels, Bathymodiolus azoricus, are sensitive to hydrostatic pressure change, but can repair DNA damage induced by retrieval from 840 m to the sea surface. In contrast, animals collected from 1700 m survived for only a few days in the laboratory, which was reflected in their poor DNA quality. These findings support the hypothesis of a physiological barrier to survival around 1000–1500 m depth, which these results show affects both vent and non-vent species alike. Based on in vitro experimental exposures to hydrogen peroxide and MMC, vent mussels appear to have sensitivities to the environmental mutagens that are not significantly different from those of coastal mussels

Carrier of reduced sulfur is a possible role for thiotaurine in symbiotic species from hydrothermal vents with thiotrophic symbionts

Experiments supporting the possible role of the free sulfur-containing amino acid thiotaurine, as a transport and storage compound for sulfide in invertebrates with thiotrophic symbionts are described. The free-living chemotrophic sulfur-oxidising bacterium, Thiobacillus hydrothermalis (strain DSMZ 7121), was used as a model for the symbionts as the actual symbionts have not been obtained in culture. Thiotaurine contains two sulfur atoms, namely the inner sulfone and the outer sulfane sulfur; the latter presents a potential source of reducing equivalents for the symbiont. Nevertheless, we observed no oxidation of thiotaurine when this compound was added to a culture of T. hydrothermalis pre-grown on sulfide. In contrast, when thiotaurine was added to the culture together with an extract of the trophosome of a vestimentiferan tubeworm from the Manus basin, we observed that thiotaurine was oxidised to hypotaurine with concomitant acidification and formation of bacterial biomass. Thus, the trophosome contains an unknown catalytic factor. We suggest that thiotaurine requires reduction prior to oxidation by T. hydrothermalisand that the host may catalyse the conversion of thiotaurine through the glutathione redox couple. This way, the host can accurately control energy delivery (as reduced sulfur) to the symbionts and can therefore control their symbiont biomass

Thiotaurine is a biomarker of sulfide-based symbiosis in deep-sea bivalves

A simple biochemical approach for demonstrating the presence of symbionts in deep-sea bivalves and for discriminating between thiotrophic and methylotrophic symbioses is described. Correspondence analysis (CA) of the free amino compound composition of nine bivalve species living in hydrothermal vents and cold seeps successfully discriminates symbiotic species from nonsymbiotic ones, and sulfuroxidizing symbionts from methylotrophic symbionts. CA was also used to infer the metabolism of Bathymodiolus azoricus, B. boomerang, and two new species of Mytilid and Vesicomyid from the Barbados. These results were consistent with the evidence obtained by ultrastructural observations of the gills and enzymatic studies, and show that CA of amino acid profiles might be a useful tool to determine the type of endosymbionts present in deep-sea bivalves. Among all the free amino acids, thiotaurine appears as the main discriminating one and is proposed as a biomarker of sulfide-based endosymbiosis in deep-sea bivalves from hydrothermal vents and cold seeps

Marine invertebrate eco-genotoxicology: a methodological overview

The last 25 years have seen major advances in the field of mammalian genotoxicology, particularly with the advent of molecular methods, some of which have spilled over into the relatively new field of eco-genotoxicology, which aims to evaluate the impact of contaminants on the natural biota. Unlike mammalian genotoxicology, where the focus is centred on a limited number of model species, efforts in the marine field have generally lacked coordination and focus, with the result that progress has been somewhat slow and fragmented. However, it is recognized that at the DNA and chromosome levels, marine invertebrates express qualitatively similar types of induced damage to that found in higher organisms (e.g. point mutations, strand breaks and chromosomal aberrations). Given that many of these species (bivalve molluscs, crustaceans, polychaete worms, etc.) are linked directly or indirectly to the human food chain, this is an important reason why one should be concerned about their exposure to environmental mutagens and carcinogens, particularly as many of these organisms have the capacity to (i) transform these agents to biologically active metabolites and (ii) accumulate toxicants in their cells and tissues at concentrations several orders of magnitude above that found in the environment. This review covers the advantages and limitations of those cytogenetic and molecular assays that have been used to address the question of genotoxicity in the cells and early life stages of selected marine invertebrate species. It concludes with the recommendation for the adoption of standardized test procedures, leading to a tiered approach in future eco-genotoxicity testing