VIVALDI, A H2020 European Project aiming at preventing and mitigating farmed bivalve diseases

The European shellfish industry enjoys a privileged position on the global scene. Its social dimension is essential, as it employs a high number of people in more than 8000 companies, mostly micro-companies. Shellfish production in Europe is little diversified and mainly relies on the industrially produced mussels, oysters and clams. Over the recent years, this sector has grown more slowly than other fish farming sectors, notably because it depends a great deal on the environmental quality and the emergence of diseases. Mortality events, linked to pathogen organisms such as viruses, bacteria and parasites (protozoa), tend to weaken the production’s sustainability. In this context, the European project VIVALDI (PreVenting and mItigating farmed biVALve DIseases) aims at increasing the sustainability and competitiveness of the shellfish industry in Europe, developing tools and approaches with a view to better preventing and controlling marine bivalve diseases. VIVALDI is a 4-years European Horizon 2020 project coordinated by Ifremer (2016-2020): 21 mostly European, public and private partners are involved, representing the diversity of the European shellfish industry landscape

Disruption of amylase genes by RNA interference affects reproduction in the Pacific oyster Crassostrea gigas

Feeding strategies and digestive capacities can have important implications for variation in energetic pathways associated with ecological and economically important traits, such as growth or reproduction in bivalve species. Here, we investigated the role of amylase in the digestive processes of Crassostrea gigas, using in vivo RNA interference. This approach also allowed us to investigate the relationship between energy intake by feeding and gametogenesis in oysters. Double-stranded (ds)RNA designed to target the two α-amylase genes A and B was injected in vivo into the visceral mass of oysters at two doses. These treatments caused significant reductions in mean mRNA levels of the amylase genes: −50.7% and −59% mRNA A, and −71.9% and −70.6% mRNA B in 15 and 75 µg dsRNA-injected oysters, respectively, relative to controls. Interestingly, reproductive knock-down phenotypes were observed for both sexes at 48 days post-injection, with a significant reduction of the gonad area (−22.5% relative to controls) and germ cell under-proliferation revealed by histology. In response to the higher dose of dsRNA, we also observed reductions in amylase activity (−53%) and absorption efficiency (−5%). Based on these data, dynamic energy budget modeling showed that the limitation of energy intake by feeding that was induced by injection of amylase dsRNA was insufficient to affect gonadic development at the level observed in the present study. This finding suggests that other driving mechanisms, such as endogenous hormonal modulation, might significantly change energy allocation to reproduction, and increase the maintenance rate in oysters in response to dsRNA injection.

Age-dependence of cultured pearl grade and colour in the black-lipped pearl oyster Pinctada margaritifera

Pinctada margaritifera is an economically important marine bivalve species for cultured pearl production in French Polynesian aquaculture. In order to evaluate the influence of donor oyster age on pearl quality traits, experiments were conducted over 6 years using both grafts and surgreffe operations. At harvest, 6 pearl quality traits were recorded and compared: surface defects, luster, grade, darkness level, and visual color. Analyzing the quality traits of pearls harvested in the initial graft process and those of pearls obtained from surgreffe experiments allowed a comparison of the influence of pearl sac cells originating from the initial mantle graft, which aged together with their recipient oysters. The results demonstrated a significant decrease between these successive grafts in luster, grade (A-B-C,) darkness level, and green color – traits that are of major importance in the pearl market. The duplicated graft experiment allowed the comparison of donor oyster families at 2 and 5 years old, where a mantle graft was inserted into recipient oysters aged 2.5 years old. The results showed the same tendencies to a lesser extent, with 1) an improved pearl grade, predominantly through a most important rate of 0 surface defect category, and 2) a green / grey ratio in favor of the younger donor. A comparison between the graft-surgreffe and the duplicated graft experiments also highlighted: 1) the indirect role played by the younger recipient oysters, which must be optimized for optimal pearl quality realization, and 2) the complex interplay between donor and recipient oysters.

Identifying Robust Proxies of Gonad Maturation for the Protandrous Hermaphrodite Tridacna maxima (Röding, 1798, Bivalvia) from Individual to Population Scale

Monitoring gonadmaturation for protandrous and functional hermaphrodite species such as the giant clamTridacna maxima is difficult due to the juxtaposition and relative proportion ofmale and female tissues in the gonad [gonadal sex ratio (GSR)]. Here, the relevance of the widely used gonadosomatic index (GSI) as proxy of giant clam gonad maturation is tested with a large dataset (n = 265). Gonadosomatic index is compared with other indices, namely the proportion of the male part harboring spermatozoids, the proportion of empty oocyte follicles, the mean oocyte diameter, and the oocyte elongation. At gonad scale, high index variability highlighted partial spawning. At individual scale, male and female maturation proxies were contrasted, showing either asynchronous emissions of male and female gametes or contrasted spermatogenesis and oogenesis duration. The GSI was mostly driven by the number and diameter of oocytes and therefore it is recommended here as primary proxy for female maturity. Except for the oocyte elongation, all indices were affected by the GSR, which ruled out drawing conclusions at population scale. These results highlight the need for maturation stage proxies that are optimized for functional hermaphrodite species.

Bacterial diseases in marine bivalves

Bivalve aquaculture is seriously affected by many bacterial pathogens that cause high losses in hatcheries as well as in natural beds. A number of Vibrio species, but also members of the genera Nocardia and Roseovarius, are considered important pathogens in aquaculture. The present work provides an updated overview of main diseases and implicated bacterial species affecting bivalves. This review focuses on aetiological agents, their diversity and virulence factors, the diagnostic methods available as well as information on the dynamics of the host-parasite relationship.

Informing Marine Spatial Planning (MSP) with numerical modelling: A case-study on shellfish aquaculture in Malpeque Bay (Eastern Canada)

A moratorium on further bivalve leasing was established in 1999–2000 in Prince Edward Island (Canada). Recently, a marine spatial planning process was initiated explore potential mussel culture expansion in Malpeque Bay. This study focuses on the effects of a projected expansion scenario on productivity of existing leases and available suspended food resources. The aim is to provide a robust scientific assessment using available datasets and three modelling approaches ranging in complexity: (1) a connectivity analysis among culture areas; (2) a scenario analysis of organic seston dynamics based on a simplified biogeochemical model; and (3) a scenario analysis of phytoplankton dynamics based on an ecosystem model. These complementary approaches suggest (1) new leases can affect existing culture both through direct connectivity and through bay-scale effects driven by the overall increase in mussel biomass, and (2) a net reduction of phytoplankton within the bounds of its natural variation in the area.

Report of the Workshop on Growth-increment Chronologies in Marine Fish: climate-ecosystem interactions in the North Atlantic 2 (WKGIC2). 18-22 April 2016 Esporles, Mallorca, Spain

Over the past several decades, thousands of otoliths, bivalve shells, and scales have been collected for the purposes of age determination and remain archived in European and North American fisheries laboratories. Advances in digital imaging and computer software combined with techniques developed by tree-ring scientists provide a means by which to extract additional levels of information in these calcified structures and generate annually resolved (one value per year), multidecadal time-series of population-level growth anomalies. Chemical and isotopic properties may also be extracted to provide additional information regarding the environmental conditions these organisms experienced.Given that they are exactly placed in time, chronologies can be directly compared to instrumental climate records, chronologies from other regions or species, or time-seriesof other biological phenomena. In this way, chronologies may be used to reconstruct historical ranges of environmental variability, identify climatic drivers of growth, establish linkages within and among species, and generate ecosystem-level indicators. Following the first workshop in Hamburg, Germany, in December 2014, the second workshop on Growth increment Chronologies in Marine Fish: climate-ecosystem interactions in the North Atlantic (WKGIC2) met at the Mediterranean Institute for Advanced Studies headquarters in Esporles, Spain, on 18–22 April 2016, chaired by Bryan Black (USA) and Christoph Stransky (Germany).Thirty-six participants from fifteen different countries attended. Objectives were to i) review the applications of chronologies developed from growth-increment widths in the hard parts (otoliths, shells, scales) of marine fish and bivalve species ii) review the fundamentals of crossdating and chronology development, iii) discuss assumptions and limitations of these approaches, iv) measure otolith growth-increment widths in image analysis software, v) learn software to statistically check increment dating accuracy, vi) generate a growth increment chronology and relate it to climate indices, and vii) initiate cooperative projects or training exercises to commence after the workshop.The workshop began with an overview of tree-ring techniques of chronology development, including a hands-on exercise in cross dating. Next, we discussed the applications of fish and bivalve biochronologies and the range of issues that could be addressed. We then reviewed key assumptions and limitations, especially those associated with short-lived species for which there are numerous and extensive otolith archives in European fisheries labs. Next, participants were provided with images of European plaice otoliths from the North Sea and taught to measure increment widths in image analysis software. Upon completion of measurements, techniques of chronology development were discussed and contrasted to those that have been applied for long-lived species. Plaice growth time-series were then related to environmental variability using the KNMI Climate Explorer. Finally, potential future collaborations and funding opportunities were discussed, and there was a clear desire to meet again to compare various statistical techniques for chronology development using a range existing fish, bivalve, and tree growth-increment datasets. Overall, we hope to increase the use of these techniques, and over the long term, develop networks of biochronologies for integrative analyses of ecosystem functioning and relationships to long-term climate variability and fishing pressure.

Determination of thresholds in marine molluscs as an alternative to the Environmental Quality Standards in marine water defined in the Water Framework Directive

The Water Framework Directive (WFD) establishes Environmental Quality Standards (EQS) in marine water for 34 priority substances. Among these substances, 25 are hydrophobic and bioaccumulable (2 metals and 23 organic compounds). For these 25 substances, monitoring in water matrix is not appropriate and an alternative matrix should be developed. Bivalve mollusks, particularly mussels (Mytilus edulis, Mytilus galloprovincialis), are used by Ifremer as a quantitative biological indicator since 1979 in France, to assess the marine water quality. This study has been carried out in order to determine thresholds in mussels at least as protective as EQS in marine water laid down by the WFD. Three steps are defined: - Provide an overview of knowledges about the relations between the concentrations of contaminants in the marine water and mussels through bioaccumulation factor (BAF) and bioconcentration factor (BCF). This allows to examine how a BCF or a BAF can be determined: BCF can be determined experimentally (according to US EPA or ASTM standards), or by Quantitative Activity-Structure Relationship models (QSAR): four equations can be used for mussels. BAF can be determined by field experiment; but none standards exists. It could be determined by using QSAR but this method is considered as invalid for mussels, or by using existing model: Dynamic Budget Model, but this is complex to use. - Collect concentrations data in marine water (Cwater) in bibliography for those 25 substances; and compare them with concentration in mussels (Cmussels) obtained through French monitoring network of chemicals contaminants (ROCCH) and biological integrator network RINBIO. According to available data, this leads to determine the BAF or the BCF (Cmussels /Cwater) with field data. - Compare BAF and BCF values (when available) obtained with various methods for these substances: BCF (stemming from the bibliography, using experimental process), BCF calculated by QSAR and BAF determined using field data. This study points out that experimental BCF data are available for 3 substances (Chlorpyrifos, HCH, Pentachlorobenzene). BCF by QSAR can be calculated for 20 substances. The use of field data allows to evaluate 4 BAF for organic compounds and 2 BAF for metals. Using these BAF or BCF value, thresholds in shellfish can be determined as an alternative to EQS in marine water.