Search

Abstract Background The shell of the pearl-producing bivalve Pinctada margaritifera is composed of an organic cell-free matrix that plays a key role in the dynamic process of biologically-controlled biomineralization. In order to increase genomic resources and identify shell matrix proteins implicated in biomineralization in P. margaritifera, high-throughput Expressed Sequence Tag (EST) pyrosequencing was undertaken on the calcifying mantle, combined with a proteomic analysis of the shell. Results We report the functional analysis of 276 738 sequences, leading to the constitution of an unprecedented catalog of 82 P. margaritifera biomineralization-related mantle protein sequences. Components of the current "chitin-silk fibroin gel-acidic macromolecule" model of biomineralization processes were found, in particular a homolog of a biomineralization protein (Pif-177) recently discovered in P. fucata. Among these sequences, we could show the localization of two other biomineralization protein transcripts, pmarg-aspein and pmarg-pearlin, in two distinct areas of the outer mantle epithelium, suggesting their implication in calcite and aragonite formation. Finally, by combining the EST approach with a proteomic mass spectrometry analysis of proteins isolated from the P. margaritifera shell organic matrix, we demonstrated the presence of 30 sequences containing almost all of the shell proteins that have been previously described from shell matrix protein analyses of the Pinctada genus. The integration of these two methods allowed the global composition of biomineralizing tissue and calcified structures to be examined in tandem for the first time. Conclusions This EST study made on the calcifying tissue of P. margaritifera is the first description of pyrosequencing on a pearl-producing bivalve species. Our results provide direct evidence that our EST data set covers most of the diversity of the matrix protein of P. margaritifera shell, but also that the mantle transcripts encode proteins present in P. margaritifera shell, hence demonstrating their implication in shell formation. Combining transcriptomic and proteomic approaches is therefore a powerful way to identify proteins involved in biomineralization. Data generated in this study supply the most comprehensive list of biomineralization-related sequences presently available among protostomian species, and represent a major breakthrough in the field of molluskan biomineralization.

The evolution of eutrophication (i.e., phytoplankton biomass) during recent decades was examined in the coastal waters of Vilaine Bay (VB, France) in relation to those in their main external nutrient sources, the Loire and Vilaine Rivers. Dynamic Linear Models, corroborated by the Mann-Kendall test, were used to study long-term trends and seasonality of dissolved inorganic nutrient and chlorophyll a concentrations (Chl a) in rivers and coastal waters between 1980 and 2013. The reduction in dissolved riverine inorganic phosphorus concentrations (DIP) from the early 1990s led to the decrease in their Chl a levels. However, dissolved inorganic nitrogen concentrations (DIN) decreased only slightly in the Vilaine and actually increased in the Loire, especially during summer. Simultaneously, phytoplankton in the VB has undergone profound changes with: (1) increase in biomass, (2) change in the position of the annual peak from spring to summer, and (3) increase in diatom:dinoflagellate ratios, especially in summer. The increase in phytoplankton biomass in VB, particularly in summer, was probably due to increased DIN loads from the Loire, sustained by internal regeneration of DIP and dissolved silicate from sediments. This long-term ecosystem-scale analysis reports the consequence of nutrient management scenarios focused solely on P reduction. Freshwater ecosystems upstream reveal successful recoveries through the control of P alone, while eutrophication continues to increase downstream, especially during the period of N limitation. Therefore, nutrient management strategies, paying particular attention to diffuse N-sources, are required to control eutrophication in receiving coastal waters. [ABSTRACT FROM AUTHOR]

Producing high quality cultured black pearls from Pinctada margaritifera is one of the major challenges for the “pearl oyster” industry in French Polynesia. In order to assess donor effect on cultured pearl quality, wild Pinctada margaritifera originating from the Tuamotu Archipelago were used in a duplicated grafting experiment. After 12 months of culture, nucleus retention was assessed and seven pearl quality traits recorded on the 454 cultured pearls harvested from the experiment. The traits scored were nacre thickness and pearl weight, surface defects, lustre, grade, and the colour components: 1) darkness of cultured pearl colour, and 2) visual perception of colour class (bodycolor and/or overtone). Our results demonstrate for the first time that individual wild donors of implanted mantle grafts significantly affect these seven quality traits in P. margaritifera cultured pearls. This finding was repeated in two series of grafts made by different professional grafters. The wild donors could be ranked from “best” (e.g., the donor whose grafts produced the cultured pearl with the maximum lustre) to the “worst”. Moreover, we showed strong correlations between: 1) cultured pearl nacre thickness and grade, with grade A showing the greatest nacre thickness on average compared with grade D and rejects; and 2) nacre thickness/cultured pearl weight and colour components (darkness and visual “colour categories”), with the palest cultured pearls (i.e. white cultured pearls) being the smallest (lowest nacre thickness and weight). Thus, one way of enhancing P. margaritifera foundation stocks for a selective breeding program could be to select the “best” donors, using appropriate molecular tools. Generation of selected donor lines from these stocks through hatchery production would be one way to increase the quality of cultured pearl farming of P. margaritifera in French Polynesia. [ABSTRACT FROM AUTHOR]

Pinctada margaritifera mollusc is cultivated in French Polynesia for the production of black pearls. For this study, the colour of juvenile samples (48 days old) was investigated in the visible range spectra (430–700 nm) using spectrophotometry. A first measurement was done with the soft parts still inside the shell (entire animal). Then, the soft parts were removed in order to do a second measurement on the growing edge of the shell. Comparison of the two measurements shows that the estimation of the living animal colour with unaided eye is strongly influenced by the colour of the soft parts. The use of the International Commission on Illumination (ISI) chromaticity diagram shows that at this growth stage, the shells are “white”; i.e. present no absorptions to the visible part. Multivariate statistical analyses based on the intensities of 6 wavelengths show that the shell colour is less variable than the colours of the entire animals. Wavelength intensities of the shells are similar, so no colour trend is visible. On the other hand, the colours of the entire animals are darker, and more variable. At this growth stage, the shell colour is not predictable for a potential selection of receiver or donor. [ABSTRACT FROM PUBLISHER]

Pearl oyster grafting is a complex surgical operation that should lead to pearl formation after approximately eighteen months. Although this technique has been used for many years in French Polynesia, the grafting process is still not standardised. While studies have been carried out in order to improve graft performance and yield, these remain highly variable due to post-grafting mortality, nucleus rejection and unreliable pearl quality, all of which constrain pearl farm profitability. The present study uses histological analysis to monitor oysters that either rejected or retained their nuclei. Both groups of oysters are compared in terms of evolution of the graft, which could influence retention, and the development of a pearl sac in cases where grafting was successful. Data show that rejection phenomena are linked to a number of causes, notably an inflammatory reaction in the "receiving" oyster, the presence of numerous tissue lesions and the quality of the grafted tissue. These results suggest that study is needed on the different concomitant elements of the grafting process: the graft "donor" oysters, the nucleus and the "receiving" oyster and their interactions. [ABSTRACT FROM AUTHOR]

We assayed European flat oyster, Ostrea edulis, hemocyte parameters, circulating and tissue-infiltrating hemocyte densities, circulating hemocyte type distribution and lysosomal enzyme contents, to possibly relate these hematological parameters to Bonamia ostreae infection. Circulating hemocyte densities were not statistically different between infected and uninfected oysters. In contrast, the number of tissue-infiltrating hemocytes increased with infection intensity suggesting a recruitment process at the site of infection and a possibility for cells to migrate from circulatory system to connective tissues. Lysosomal enzymes were localized mainly in granulocytes both infected and uninfected, and mean of α-naphtyl butyrate esterase activity decreased with increasing B. ostreae infection level. The main response observed was a change in hemocyte type distribution between uninfected and infected oysters and greater tissue-infiltrating hemocytes with increased infections. These results suggest that the decrease of circulating granulocytes, and, consequently of some cell enzyme activities may be related with B. ostreae infection. [Copyright &y& Elsevier]

Abstract As human population increases worldwide, water quality will become increasingly problematic, and food consumed raw may be of higher risk. This is already evident for oysters grown in coastal areas - despite regulations based on bacterial indicators, oysters are still implicated in food-borne outbreaks worldwide. The pathogens most frequently detected are human noroviruses, which are shed at high concentrations in human excreta and are very resistant to environmental conditions. Sewage treatment plants usually apply a variety of steps such as activated sludge treatment, chlorine or UV disinfection to eliminate contaminants, these processes have variable efficacy. This study demonstrates the impact of replacing an old lagoon-based sewage treatment plant with a new membrane bioreactor sewage treatment plant on human norovirus levels in treated sewage and oysters. While comparable norovirus concentrations were detected in the influent samples, a clear difference was observed in effluent quality, as norovirus was only detected in one sample after treatment in the new membrane bioreactor system, confirming the efficiency of such technology. As a direct impact, oysters located close to the membrane bioreactor sewage outfall were less frequently contaminated by norovirus, and showed lower concentrations compared to the first period of the study when they were exposed to sewage effluent from the lagoon outfall. Shellfish located upstream showed comparable contamination levels suggesting that there are also other sources of norovirus contamination in the estuary. Considering the health benefits of shellfish consumption, improving wastewater quality will make an important contribution to enhancing the safety of shellfish and international food security. Highlights • Old lagoon sewage treatment plant is inefficient in eliminating human norovirus. • Membrane bioreactor treatment plant improves the microbiological quality of effluent. • Oyster contamination by norovirus can be prevented by improving sewage treatment. • Several factors may contribute to water contamination in an estuary. • Lowering human sewage input in surface waters will be beneficial for international food security. [ABSTRACT FROM AUTHOR]

The protistan parasite Mikrocytos mackini, the causative agent of Denman Island disease in the oyster Crassostrea gigas in British Columbia, Canada, is of wide concern because it can infect other oyster species and because its life cycle, mode of transmission, and origins are unknown. PCR and fluorescent in situ hybridization (FISH) assays were developed for M. mackini, the PCR assay was validated against standard histopathological diagnosis, and a preliminary phylogenetic analysis of the M. mackini small-subunit ribosomal RNA gene (SSU rDNA) was undertaken. A PCR designed specifically not to amplify host DNA generated a 544 bp SSU rDNA fragment from M. mackini-infected oysters and enriched M. mackini cell isolates, but not from uninfected control oysters. This fragment was confirmed by FISH to be M. mackini SSU rDNA. A M. mackini-specific PCR was then designed which detected 3 to 4x more M. mackini infections in 1056 wild oysters from Denman Island, British Columbia, than standard histopathology. Mikrocytos mackini prevalence estimates based on both PCR and histopathology increased (PCR from 4.4 to 7.4%, histopathology from 1.2 to 2.1%) when gross lesions were processed in addition to standard samples (i.e. transverse sections for histopathology, left outer palp DNA for PCR). The use of histopathology and tissue imprints plus PCR, and standard samples plus observed gross lesions, represented a 'total evidence' approach that provided the most realistic estimates of the true prevalence of M. mackini. Maximum parsimony and evolutionary distance phylogenetic analyses suggested that M. mackini may be a basal eukaryote, although it is not closely related to other known protistan taxa.