Spatio-temporal variability of trace elements fingerprints in cockle (Cerastoderma edule) shells and its relevance for tracing geographic origin.

Understanding spatio-temporal variability of trace elements fingerprints (TEF) in bivalve shells is paramount to determine the discrimination power of this analytical approach and secure traceability along supply chains. Spatio-temporal variability of TEF was assessed in cockle (Cerastoderma edule) shells using inductively coupled plasma-mass spectrometry (ICP-MS). Four elemental ratios (Mg/Ca, Mn/Ca, Sr/Ca and Ba/Ca) were measured from the shells of specimens originating from eight different ecosystems along the Portuguese coast, as well as from four different areas, within one of them, over two consecutive years (2013 and 2014). TEF varied significantly in the shells of bivalves originating from the eight ecosystems surveyed in the present study. Linear discriminant function analyses assigned sampled cockles to each of the eight ecosystems with an average accuracy of 90%. Elemental ratios also displayed significant differences between the two consecutive years in the four areas monitored in the same ecosystem. Overall, while TEF displayed by cockle shells can be successfully used to trace their geographic origin, a periodical verification of TEF (>6 months and <1 year) is required to control for temporal variability whenever comparing specimens originating from the same area collected more than six months apart.

Effects of sample storage and shell orientation on LA-ICPMS trace element measurements on deep-sea mussels.

Geochemical markers are being increasingly applied to fundamental questions in population and community ecology in marine habitats because they allow inferences on individuals dispersal, but vital effects, small sample size and instrumental limitation are still challenging particularly in deep-sea studies. Here we use shells of the deep-sea bivalve Idas modiolaeformis to assess potential effects of sample storage, mineralogy, and valve orientation on LA-ICPMS measurements. Trace element concentrations of (24)Mg, (43)Ca, (88)Sr, (137)Ba, (208)Pb, and (238)U are not affected by the two most commonly used storage methods of biologic deep-sea samples (frozen at -20°C and fixed in 95% ethanol); thus combined analysis of differently preserved specimens is possible when the number of individuals is insufficient and distinct sample fixation is needed for multiple purposes. Valve orientation had a strong impact on quantification of trace elements in the calcitic but not in the aragonitic layer of adult shells. Hence, to enable comparisons between adult shells and entirely aragonitic embryonic shells, a reference map of site-specific signatures can potentially be generated using the aragonitic layer of the adult shells. Understanding ontogenetic changes and environmental effects in trace element incorporation is critical before geochemical fingerprinting can be used as a tool for larval dispersal studies in the deep-sea.

Trace element fingerprinting of cockle (Cerastoderma edule) shells can reveal harvesting location in adjacent areas.

Determining seafood geographic origin is critical for controlling its quality and safeguarding the interest of consumers. Here, we use trace element fingerprinting (TEF) of bivalve shells to discriminate the geographic origin of specimens. Barium (Ba), manganese (Mn), magnesium (Mg), strontium (Sr) and lead (Pb) were quantified in cockle shells (Cerastoderma edule) captured with two fishing methods (by hand and by hand-raking) and from five adjacent fishing locations within an estuarine system (Ria de Aveiro, Portugal). Results suggest no differences in TEF of cockle shells captured by hand or by hand-raking, thus confirming that metal rakes do not act as a potential source of metal contamination that could somehow bias TEF results. In contrast, significant differences were recorded among locations for all trace elements analysed. A Canonical Analysis of Principal Coordinates (CAP) revealed that 92% of the samples could be successfully classified according to their fishing location using TEF. We show that TEF can be an accurate, fast and reliable method to determine the geographic origin of bivalves, even among locations separated less than 1 km apart within the same estuarine system. Nonetheless, follow up studies are needed to determine if TEF can reliably discriminate between bivalves originating from different ecosystems.

Are organic falls bridging reduced environments in the deep sea? - results from colonization experiments in the Gulf of Cádiz.

Organic falls create localised patches of organic enrichment and disturbance where enhanced degradation is mediated by diversified microbial assemblages and specialized fauna. The view of organic falls as "stepping stones" for the colonization of deep-sea reducing environments has been often loosely used, but much remains to be proven concerning their capability to bridge dispersal among such environments. Aiming the clarification of this issue, we used an experimental approach to answer the following questions: Are relatively small organic falls in the deep sea capable of sustaining taxonomically and trophically diverse assemblages over demographically relevant temporal scales? Are there important depth- or site-related sources of variability for the composition and structure of these assemblages? Is the proximity of other reducing environments influential for their colonization? We analysed the taxonomical and trophic diversity patterns and partitioning (α- and ß-diversity) of the macrofaunal assemblages recruited in small colonization devices with organic and inorganic substrata after 1-2 years of deployment on mud volcanoes of the Gulf of Cádiz. Our results show that small organic falls can sustain highly diverse and trophically coherent assemblages for time periods allowing growth to reproductive maturity, and successive generations of dominant species. The composition and structure of the assemblages showed variability consistent with their biogeographic and bathymetric contexts. However, the proximity of cold seeps had limited influence on the similarity between the assemblages of these two habitats and organic falls sustained a distinctive fauna with dominant substrate-specific taxa. We conclude that it is unlikely that small organic falls may regularly ensure population connectivity among cold seeps and vents. They may be a recurrent source of evolutionary candidates for the colonization of such ecosystems. However, there may be a critical size of organic fall to create the necessary intense and persistent reducing conditions for sustaining typical chemosymbiotic vent and seep organisms.

A complex picture of associations between two host mussels and symbiotic bacteria in the Northeast Atlantic.

Among chemosymbiotic metazoans found at deep-sea hydrothermal vents, cold seeps and organic falls, members of the mussel clade Bathymodiolinae (Bivalvia: Mytilidae) have evolved interactions with a higher diversity of bacterial lineages than other bivalve groups. Here, we characterized the bacteria associated with "Bathymodiolus" mauritanicus and Idas-like specimens from three sites in the Northeast Atlantic (two mud volcanoes in the Gulf of Cadiz and one seamount of the Gorringe Bank). Phylogenetic analysis of bacterial 16S rRNA-encoding gene sequences demonstrated that "B". mauritanicus has a dual symbiosis dominated by two phylotypes of methane-oxidising bacteria and a less abundant phylotype of a sulphur-oxidising bacterium. The latter was the dominant phylotype in a sympatric population of Idas-like mussels at the Darwin mud volcano. These results are the first report of a bacterial phylotype shared between two deep-sea mussels from divergent clades. This sulphur-oxidising bacterium was absent from Idas-like specimens from the other two sites (Gorringe Bank and Meknès mud volcano), in which bacterial clone libraries were dominated by other Gammaproteobacteria related to symbionts previously identified in Idas modiolaeformis from the Eastern Mediterranean. All Idas-like specimens studied herein are closely related and also related to I. modiolaeformis. However, they probably display different associations with bacteria, with the possible absence of both methane- and sulphur-oxidising symbionts at the Gorringe Bank. These results draw a very complex picture of associations between mussels and bacteria in the Northeast Atlantic, which could be highly variable depending on locale characteristics of the habitats.