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.

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.

Distribuição e abundância da malacofauna epibentônica no Parracho de Maracajaú, RN, Brasil

The mollusks present a great taxon variety and life habits in coral reefs being good environmental indicators. It is important to know the distribution pattern of the mollusks and the processes that influence it, so that disturbances in sea ecosystems could be monitored. The present study aims to accomplish the inventory and distribution of epibenthic mollusks in the Parracho of Maracajaú. 23 sites in different habitats of the Parracho were settled: 11 in the reef habitat, 3 in the sandy bottom and 9 in the seagrass bed. Qualitative and quantitative samplings have been done through snorkeling and scuba diving. Three band transects (10m²) were sampled in each site and the data were obtained to each m² of the transect, where the species were counted and the environmental variables (rugosity and recovery of the substratum) were valued. The data were submitted to multivariate analyses in order to find possible distribution patterns that could be associated to the substratum variables. The diversity indexes were calculated for each reef sites and compared with each other. A number of 46 species were registered. The reef habit at should to be the richnest area while the sandy bottom was poorest one. In the reef habitat, the mollusks were associated to rugosity and recovering of frondose algae and zoanthids, while for the seagrass bed, the animals exhibited a richness variation associated to the muddy and sandy sediment. There were found 3 species economically explored, what requires an appropriate management for the maintenance and conservation of the area resources in a sustainable way

Why is Asari (= Manila) clam Ruditapes philippinarum fitness poor in Arcachon Bay: a meta-analysis to answer?

Asari (= Manila) clam, Ruditapes philippinarum, is the second bivalve mollusc in terms of production in the world and, in many coastal areas, can beget important socio-economic issues. In Europe, this species was introduced after 1973. In Arcachon Bay, after a decade of aquaculture attempt, Asari clam rapidly constituted neo-naturalized population which is now fished. However, recent studies emphasized the decline of population and individual performances. In the framework of a national project (REPAMEP), some elements of fitness, stressors and responses in Arcachon bay were measured and compared to international data (41 publications, 9 countries). The condition index (CI=flesh weight/shell weight) was the lowest among all compared sites. Variation in average Chla concentration explained 30% of variation of CI among different areas. Among potential diseases, perkinsosis was particularly prevalent in Arcachon Bay, with high abundance, and Asari clams underwent Brown Muscle Disease, a pathology strictly restricted to this lagoon. Overall element contamination was relatively low, although arsenic, cobalt, nickel and chromium displayed higher values than in other ecosystems where Asari clam is exploited. Finally, total hemocyte count (THC) of Asari clam in Arcachon Bay, related to the immune system activity, exhibited values that were also under what is generally observed elsewhere. In conclusion, this study, with all reserves due to heterogeneity of available data, suggest that the particularly low fitness of Asari clam in Arcachon Bay is due to poor trophic condition, high prevalence and intensity of a disease (perkinsosis), moderate inorganic contamination, and poor efficiency of the immune system.

Distribuição espaço-temporal e dieta de Lontra longicaudis (Carnivora: mustelidae) em região costeira do sul do RS

A lontra-neotropical (Lontra longicaudis) é um carnívoro semi-aquático, com adaptações morfológicas para viver nos mais diversos habitats aquáticos, como rios, lagos, mangues e estuários. Além disso, também é encontrada em ambientes marinhos, onde se alimenta, ou apenas transita. São carnívoros que se alimentam principalmente de peixes e crustáceos. O objetivo desde trabalho foi verificar a utilização de ambientes de influência do mar, por L. longicaudis, no litoral sul do RS. A área de estudo foi a Praia do Cassino, onde foram percorridos seis cursos d’água (sangradouros), por cerca de 1 km em cada, à procura de fezes de lontras, entre dezembro de 2009 e novembro de 2010. As fezes foram analisadas para determinar a distribuição espaço-temporal e a dieta das lontras. Foram encontradas 75 fezes de lontras, sendo a maior quantidade no inverno e outono, diminuindo na primavera e verão. As maiores quantidades de fezes foram encontradas nos sangradouros R7 e R9, por estes serem mais extensos e profundos. As menores quantidades de fezes nos sangradouros R4, R8 e R10 se deve ao fato de estes serem menores e menos profundos. Os peixes foram as principais presas das lontras, seguidos pelos crustáceos, anfíbios, moluscos, insetos, aves e mamíferos. Os peixes foram mais predados na maior parte das estações, exceto no outono, quando os crustáceos predominaram. No inverno, os anfíbios predominaram sobre os crustáceos, sendo o segundo grupo mais predado. Os peixes mais consumidos foram Perciformes e Siluriformes. Foi verificado que as lontras utilizam os sangradouros da Praia do Cassino, mesmo estes não possuindo vegetação e substrato mais favoráveis à espécie. A maior utilização dos ambientes durante o inverno provavelmente se deve ao fato de neste período os sangradouros estarem mais profundos. A dieta das lontras variou ao longo do ano, possivelmente conforme a disponibilidade das presas.

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.

Aspects of the biology of Lutraria lutraria (L.) (bivalvia : Mactracea)

The life history of a population of Lutraria lutraria in a depth of 7m at Hunterston, Ayrshire is discussed. Much of the present population Is thought to have settled in 1967. The functional morphology of Lutraria is described and related to its life as a large, deep-burrowing bivalve. Lutraria spawned in late spring and continued to do so through the summer in 1979 and 1980. Animals became spent in August and September. Unsuccessful attempts were made to induce spawning in the laboratory. Artificial fertilization was successful but development did not proceed beyond the ciliated gastrula stage. Larvae of Lutraria were not identified in plankton samples and young stages were not encountered in sieved sediment samples. The biochemical cycle of the total animal and five component parts (gonad and visceral mass, digestive gland, adductor muscle, siphon and 'other' tissue) is investigated. A marked increase in weight, reflected in an increase in weight of the component parts, was recorded in Autumn 1979. This is thought to be related to an exceptional increase in the phytoplankton at this time. Although a relationship between the biochemical cycle and reproductive cycle remains uncertain, definite seasonal changes were recorded in the respiration rate of Lutraria. At 10°C, the maximum rate of a standard 20g animal was 0.1283m1s 02/g. dry wt./hr. in May 1980 and the minimum rate was 0.O59mls 02/g. dry wt./hr. in October 1980. The effect of temperature on respiration rate was also investigated. Significant differences were recorded for five experimental temperatures (10°C, 15°C, 20°C, 25°C and 30 °C) in August and October but only between two temperatures (10 C and 30 C) in April. There was a decrease in respiration rate at 30 C in August and October, but an increase in April. Respiration rate is affected by a reduction in oxygen tension. A variety of responses were recorded with a small degree of regulation shown. Individuals of Lutraria were able to survive 48 hours under anaerobic conditions. In fully oxygenated conditions heart rate ranged from 4-15 beats per minute with an average of 8 beats per minute. Heart beat was markedly affected by changes in temperature and oxygen tension, increasing to a maximum 22 beats per minute at 25 C, and decreasing to a minimum 2 beats per minute in anaerobic conditions. Heart rate is reduced (12 beats per minute to 5 beats per minute) on exposure to air. Lutraria exhibits an intermittent pattern of pumping activity. Under normal conditions 35% of the time is spent pumping and this Increases as oxygen is reduced (3.00mls 02/litre) to 65% of the time spent pumping. 15. Under normal conditions the respiratory flow varies between 0.382 litres per hour and 1.023 litres per hxir. Adult Lutraria maintain their ability to burrow, albeit slowly.

Site n°5. The Pertuis Breton (Bay of Biscay) France

1.This report presents the results of a field study conducted in the ECASA test site nOS in the Pertuis Breton, France. The site is located on the Atlantic West coasts. It is open to the bay of Biscay, but is slightly protected against westerly winds. The bay has been exploited by intertidal mussels culture for centuries. 2. Within the bay, mussels (Mytilus edulis) are cultivated either by the traditional pole technique, around the bay or on longlines in the centre of the bay. The area occupied by these longline s represents 250 ha, and the resulting annual production is 1 000 tonnes of mussels. The average depth at mid tide is of 13.8 m. The sediment is sandy, with a small fraction of mud. 3. The site is subject to several regular monitoring through the local implementation of national networks aiming at protecting the environment and marine resources, on pollutants (RNO), microbiological quality of the waters (REMI), phytoplanktonic toxic species (REPHY) and growth and mortality of molluscs (REMORA). Benthic macrofauna was studied in 1976. 4. Five sampling sations were chosen along a line, starting under the longlines, and at distances of 50, 100, 200, and 400 metres from the area cultivated. A reference station was chosen in a different direction at 2300 metres of the cultivated area. Sampling methods are described in the text. _Sediments were sampled for different analyses: grain size, content in organic matter, total organic carbon and nitrogen, and phytic pigments (chlorophyll a and phaeopigments). Redox were measured in cores. The macrofauna living into the sediment was also sampled. The water column was sampled for physical (temperature, transparency) and chemical parametres, including oxygen content, salinity, organic matter, dissolved nitrogen forms, phosphates and silicates. Results from benthic macrofauna surveys indicate that there were no significant differences between the different stations and the reference station, all being classified as slightly disturbed. The bay is submitted to freshwater runoffs from two adjacent rivers. 7. The sediment is slightly modified by the culture of bivalves. Total organic carbon, total nitrogen, Eh values and pheopigments were significantly higher under the trestles than in any other stations. Other stations often did not differ from the reference station. 8. The effects of shellfish culture on the water column were. However, it was observed a small decrease of the food available to the molluscs near the rearing 9. The DEB model was able to describe and predict adequately the growth of oysters, both in the Baie des Veys and in the Loch Creran. The parametres for its use in other environment are given, but a tuning of one parametre should be performed with the help of authors. 10. Among the indicators and models for use in are as of intertidal bivalve culture, it is recommended to use the sediment quality index, TOC (Total Organic Carbon), redox and pheopigments, in surficial sediment, AMBI for the macrofauna, chlorophyll a contents and nitrogen forms in the water column, and models describing the carrying capacity, filtration rate of molluscs, and a DEB model to predict the growth of molluscs.

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.

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.

Plotosus lineatus (Thunberg, 1787)

The Striped Catfish can be recognized by its striped coloration, barbels around the mouth, and its body shape which tapers to a point posteriorly. Small juveniles are black and large adults may be less distinctly striped. Plotosus lineatus can reach a maximum length of 32 cm (13 in) and about 40cm in Persian Gulf. The body is brown with cream-colored or white longitudinal bands. The most striking feature of this species is in the fins; in fact the second dorsal, caudal and anal are fused together as in eels. In the rest of the body is quite similar to a freshwater catfish: the mouth is surrounded by four pairs of barbells, four on the upper jaw and four on the lower jaw. The first dorsal and each of the pectoral fins have a highly venomous spine. They may even be fatal. Juveniles of P. lineatus form dense ball-shaped schools of about 100 fish, while adults are solitary or occur in smaller groups of around 20 and are known to hide under ledges during the day. Adult P. lineatus search and stir the sand incessantly for crustaceans, mollusks, worms, and sometimes fish. Striped eel catfish is an oviparous fish; this species has demersal eggs and planktonic larvae. This species has evolved long ampullary canals in its electrosensory organs.

Salinity influences disease-induced mortality of the oyster Crassostrea gigas and infectivity of the ostreid herpesvirus 1 (OsHV-1)

Mortality of young Pacific oysters Crassostrea gigas associated with the ostreid herpesvirus 1 (OsHV-1) is occurring worldwide. Here, we examined for the first time the effect of salinity on OsHV-1 transmission and disease-related mortality of C. gigas, as well as salinity-related effects on the pathogen itself. To obtain donors for OsHV-1 transmission, we transferred laboratory-raised oysters to an estuary during a disease outbreak and then back to the laboratory. Oysters that tested OsHV-1 positive were placed in seawater tanks (35‰, 21°C). Water from these tanks was used to infect naïve oysters in 2 experimental setups: (1) oysters acclimated or non-acclimated to a salinity of 10, 15, 25 and 35‰ and (2) oysters acclimated to a salinity of 25‰; the latter were exposed to OsHV-1 water diluted to a salinity of 10 or 25‰. The survival of oysters exposed to OsHV-1 water and acclimated to a salinity of 10‰ was >95%, compared to only 43 to 73% survival in oysters acclimated to higher salinities (Expt 1), reflecting differences in the levels of OsHV-1 DNA and viral gene expression (Expts 1 and 2). However, the survival of their non-acclimated counterparts was only 23% (Expt 2), and the levels of OsHV-1 DNA and the expression of 4 viral genes were low (Expt 1). Thus, OsHV-1 may not have been the ultimate cause of mortality in non-acclimated oysters weakened by a salinity shock. It appears that reducing disease risk by means of low salinity is unlikely in the field.

Natural recovery of the benthic nematode assemblages associated to Zostera noltii seagrass beds after physical disturbance caused by digging harvest activity

Seagrass beds are productive ecosystems that maintain high levels of biodiversity, making them susceptible to anthropogenic pressures such as bivalve harvesting. Nematodes are considered great ecological indicators as changes in their density, diversity and structure may represent changes in the environment. This experimental fieldwork aimed to assess the impact of the bivalve harvesting on the nematodes assemblage of a seagrass bed in the Mira estuary by simulating the digging activity. Two plots were subjected to the digging (D1 and D19) and two plots were control (C11 and C18). The sampling took place in five occasions: T0 – before digging; T1 – 14 days; T2 – 45 days; T3 – 75 days; and T4 – 165 days after digging. The results showed no significant difference in the nematode assemblages’ density, diversity and trophic composition between treatments and sampling times, evidencing their high tolerance for naturally stressed environments and to the level of digging they were exposed; Recuperação natural das comunidades de nematodes bentónicos associados aos povoamentos de Zostera noltii após atividade de marisqueio Resumo: As pradarias marinhas são ecossistemas produtivos que suportam elevados níveis de biodiversidade, pelo que estão sujeitos a pressões antropogénicas. Os nematodes são bons indicadores ecológicos pois respondem rapidamente a qualquer perturbação por alterações na densidade, diversidade e estrutura. Este trabalho experimental teve como finalidade o estudo da recuperação natural das comunidades de nematodes associados aos povoamentos de Zostera noltii pela simulação da atividade de marisqueio. Dois plots foram sujeitos a revolvimento (D1 e D19) e dois plots serviram como controlo (C11 e C18) e foram efetuadas amostragens em cinco ocasiões: T0 – antes do revolvimento; T1 – 14 dias; T2 – 45 dias; T3 – 75 dias; e T4 – 165 dias após revolvimento. Os resultados obtidos não mostraram diferenças significativas na diversidade, densidade e composição trófica das comunidades de nematodes entre tratamentos e tempos de amostragem, evidenciando a sua elevada tolerância a ambientes naturalmente dinâmicos e ao nível de revolvimento a que foram expostas.

Bivalves marinhos do miocénio superior (Tortoniano superior) de Cacela (Algarve, Portugal)

Dissertação de mestrado em Estudos Marinhos e Costeiros, Unidade de Ciências e Tecnologias dos Recursos Aquáticos, Univ. do Algarve, 2000