Search

Hanging baskets are rapidly being adopted in many parts of the world to grow shell- fish, especially oysters. The adoption of this method is opening up new areas for aquaculture development. However, expansion into areas that have previously been unusable has been con- troversial, due to concerns about farms being established in areas with particularly high ecologi- cal values and sensitivity. This study investigated the impact of establishing a hanging basket oys- ter farm in the Kaipara Harbour in northern New Zealand on seagrass. Aerial photographs were analysed with a geographical imaging software in combination with field sampling to detect potential impacts. The hanging basket oyster farm technology was found to have no significant overall impact on seagrass beneath the farm. However, field sampling found a narrow band directly beneath the oyster farm growout lines of less than 5% of the farm area that had lower seagrass densities and abundance compared to adjacent control zones sampled within the farm. This area was obscured in aerial photographs by farm structures. This highly localised impact is likely due to shading or scour caused by farm structures. Overall, the results indicate that hanging basket technology for shellfish aquaculture has minimal environmental impact on underlying seagrass, representing an improvement over traditional culture technologies. Furthermore, the results confirm that remote sensing methods are useful tools for examining aquaculture impacts on seagrass communities, but only when combined with field sampling.

Juvenile oyster disease (JOD) in Crassostrea virginica is caused by the marine bacterium Roseovarius crassostreae. Although the 16S rRNA genes of the bacterial isolates exhibit little variation, 2 genetic signatures (GSI and GSII) may be discerned by Ava I digestion of the 16S-23S internal transcribed spacer (ITS). In this study we analyzed isolates from JOD epizootics throughout the northeastern USA (including affected adults for the first time) to better understand how oyster populations encounter and become affected by the pathogen. Isolates from a given epizootic usually had the same ITS signature; however, the involvement of both genetic signatures was occasionally detected, even within the same oyster. Sequencing was used to localize the variable Ava I site to a 100 bp region of low sequence identity, and detection of additional base changes resulted in the identification of 11 distinct genotypes. One genotype was found only in Martha's Vineyard, Massachusetts, USA and persisted in JOD survivors. Two genotypes were associated with Maine epizootics, and both were believed to be unique to that region until 2004, when one was detected in Martha's Vineyard among oysters that had survived colonization by the local genotype. Apparent competition between those 2 genotypes was also detected among a population of juveniles. Five genotypes were found only in New York, and the other 3 were isolated from both New York and from around Cape Cod, Massachusetts. Relationships between the geographic occurrence and phylogenetic relatedness of genotypes were compared with regional current patterns to identify possible mechanisms controlling their distribution.

Oysters Crassostrea virginica are 'keystone' estuarine species in the southeastern USA, providing essential fish habitat, food for human consumption, filtration of water bodies, and protection against shoreline erosion. Relatively few oyster pathology studies have been conducted in Georgia. The parasitic protozoan Perkinsus marinus was first observed here in the 1960s, but has not been investigated since the late 1990s, when increasing oyster infection levels were apparent. In the late 1990s and early 2000s, the region suffered a prolonged drought, resulting in elevated salinities and the proliferation of various diseases in the marine environment. By 2003, salinities had returned to normal levels, but the effect of the drought on oysters was unknown. In June 2003, oyster reefs throughout Chatham County were sampled to evaluate the prevalence and intensity of P. marinus. The disease appears to have remained prevalent in the coastal waters of Georgia (100% prevalence at some sites), but the intensity was low, ranging from 0 to 1.83 on a scale where heavy infections rated a score of 5. While the disease did not occur at levels high enough to cause oyster mortalities, further monitoring, particularly on a temporal scale, is warranted.

Seed losses of Pacific oysters Crassostrea gigas have been associated with an ostreid herpesvirus-1 (OsHV-1) in Europe, and in 2002, a similar OsHV was detected in Tomales Bay, California, USA. In May of 2003, 5 stocks of seed Pacific oysters were planted at 2 sites (Inner Bay and Outer Bay) in Tomales Bay and monitored for mortality, presence/prevalence of OsHV (using polymerase chain reaction [PCR] and histology), and growth. Temperature (degrees C) and salinity data were collected every half an hour at each site. OsHV was detected at both the Inner and Outer Bay sites on the same sample date and mean temperature predicted OsHV presence (p0.005). High levels of mortality occurred 2 wk (Inner Bay site) and 4 wk (Outer Bay site) after OsHV detection. OsHV presence predicted mortality (p = 0.01). Temperature maximums and overall temperature exposure were greater at the Inner Bay site and may explain why mortality affected these oysters sooner than oysters planted at the Outer Bay site. Differences in cumulative mortality were significant among stocks (p0.0001), but not between sites (p0.05). OsHV prevalence was similar among stocks (p0.05) and between sites (p0.05). No evidence of herpesvirus-induced Cowdry type A nuclear inclusions or other pathogens were observed. Changes in tissue and cellular architecture including dilation of the digestive tubules and nuclear chromatin margination and pycnosis were observed in OsHV-infected oysters, consistent with previously observed OsHV infections. Stocks with smaller oysters had higher mortality rates than those with larger oysters; growth rate did not correlate with mortalities (p0.05). Taken together, these data suggest that the OsHV may cause or act in synergy with temperature to kill Pacific oyster seed in Tomales Bay, but further investigation of OsHV etiology in seed oysters is needed.

Culture of native flat oysters Ostrea puelchana d'Orbigny in San Antonio Bay (San Matías Gulf, Argentina) began in 1995. After elevated mortality (33%) occurred in September 1996, 18 mo after immersion, histopathological analysis and evaluation of parasitic prevalence was carried out. In October 1997, after 31 mo of cultivation, cumulative mortality was 80%, and in December of the same year, when individuals reached marketable size, mortality was 95% and culture was discontinued. The present study describes the haemocytic parasitism that affected O. puelchana, and suggests that a Bonamia sp. was the etiological agent. This parasite should be considered as a different species from Bonamia sp. detected in Australia and New Zealand until more studies are made to determine the correct taxonomy. This work constitutes the first record of this haemocyte parasite in flat oysters from the Argentinean coast.

We have developed a PCR-assay for the diagnosis of juvenile oyster disease (JOD) based on the detection of Roseovarius crassostreae directly from affected oysters. Species-specific primers are used to amplify the 16S-23S rDNA internal transcribed spacer (ITS) of R. crassostreae, and confirmation of product identity is accomplished by restriction enzyme analysis. No false positives were obtained with either closely related bacterial species or from other DNAs present in oyster samples. The assay has the potential to detect as few as 10 cells of R. crassostreae per oyster when samples are taken from the inner valve surfaces of the animal. Inclusion of material from soft body surfaces is not necessary, and may reduce sensitivity approximately 10-fold. In a JOD-affected population, a positive PCR result was obtained from all oysters from which these bacteria were subsequently cultured. The assay also detected the presence of R. crassostreae in 2 oysters from which no R. crassostreae isolates were recovered. No R. crassostreae was detected by either PCR or bacteriology in oysters from a population that was not exhibiting JOD-signs. This assay is expected to advance regional disease management efforts and provide valuable insights into the disease process and epizootiology of JOD.

Mortalities related to the detection of Ostreid Herpesvirus 1 (OsHV-1) have been previously reported in France among larvae and spat of the Pacific oyster Crassostrea gigas. Adult oysters appear less sensitive to herpesvirus infections, although OsHV-1 has been detected in adults without signs of disease or mortality. This suggests that the virus is able to persist in its host and that adult oysters may be able to control OsHV-1 infection. Little is known about antiviral substances in invertebrates. The present work concerns the research of antiviral substances in adult oyster C. gigas, where putative antiviral activities were monitored using 3 strategies: (1) in metabolites with variable polarity, (2) in peptidic extracts and (3) in crude haemolymph. In vitro antiviral assays were based on inhibition of Herpes simplex virus type 1 (HSV-1) replication in Vero cell monolayers. All extracts presented no cytotoxicity. Antiviral activity was detected in the fresh filtered haemolymph (EC50:425 microg ml(-1)) and seasonal variation of the haemolymph antiviral activity was monitored.

Populations of eastern oysters Crassostrea virginica along the east coast of North America have repeatedly experienced epizootic mass mortality due to infections by protozoan parasites, and molecular diagnostic methodologies are fast becoming more widely available for the diagnosis of protozoan diseases of oysters. In this study we applied a modified version of an existing multiplex polymerase chain reaction (PCR) for detection of the eastern oyster parasites Haplosporidium nelsoni, H. costale and Perkinsus marinus from field-collected samples. We incorporated primers for DNA quality control based on the large subunit ribosomal RNA (LSU rRNA) gene of C. virginica. The multiplex PCR (MPCR) simultaneously amplified genomic DNA of C. virginica, and cloned DNA of H. nelsoni, P. marinus and H. costale. In field trial applications, we compared the performance of the MPCR to that of the conventional diagnostic techniques of histopathological tissue examination and the Ray/Mackin fluid thioglycollate medium (RMFT) assay. A total of 530 oysters were sampled from 18 sites at 12 locations along the east coast of the United States from the Gulf of Mexico to southern New England. The modified MPCR detected 21% oysters with H. nelsoni, 2% oysters with H. costale, and 40% oysters with P. marinus infections. In comparison, histopathological examination detected H. nelsoni and H. costale infections in 6 and 0.8% oysters, respectively, and the RMFT assay detected P. marinus infection in 31% oysters. The MPCR is a more sensitive diagnostic assay for detection of H. nelsoni, H. costale, and P. marinus, and incorporation of an oyster quality control product limits false negative results.

Since 1991, herpesvirus infections have been reported among larvae and juveniles of various bivalves. Most of the studies focused on detection of viral infections of economically impor- tant species. However, the persistence of bivalve herpesviruses in the marine environment is poorly documented. The present study concerns the role of seawater parameters in Ostreid Herpesvirus 1 (OsHV-1) detection by polymerase chain reaction (PCR). Viral DNA extracted from purified particles or virions present in infected oyster larvae were detected by PCR after storage in different media at different temperatures. The lowest detection threshold was found using distilled water or Tris EDTA buffer. In seawater, the threshold was higher. The use of sterile media permitted detection of viral DNA stored over a longer period. Storage temperature also had a significant influence on detection, with lower temperatures promoting DNA detection over a longer period. In summary, water para- meters such as temperature influenced detection of OsHV-1 DNA by PCR. However, the PCR tech- nique may also be successfully applied to samples in natural seawater. Indeed, the PCR technique permitted detection of naked viral DNA at 100 ng l -1 in seawater in bioassays.

The present study was designed to quantify the potential influence of oyster cultures (Crassostrea gigas, Thunberg) on dissolved inorganic nitrogen (DIN) renewal in a shallow coastal lagoon (Thau, France), where nitrogen is the factor limiting primary production. The approach was based on a comparative in situ analysis of the seasonal patterns of nitrogenous nutrient exchanges at both the water sediment and oyster cultures water interfaces. The experiments were performed at a station under the influence of oyster cultures (UC) compared with benthic exchanges in a station outside their direct influence (OC) which was used as the reference site for benthic fluxes. In contrast to what it is usually reported for shallow ecosystems, I found evidence of a key influence of sus- pended oyster culture units (OCU) on the regeneration in the water column of nitrogen available to the phytoplankton, with benthic remineralisation being only of secondary importance. The results also showed that the greatest impact of oyster cultures was on DIN renewal (e.g. nitrogen transfor- mation) in the water column, not on particulate organic nitrogen transfer to the sediment. In the sec- tor studied, it also appeared that nitrogen recycling by both sediments and OCU could satisfy the nitrogen requirements of the primary producers except in summer when other internal nitrogen sources were necessary. Conversely, during most seasons, pelagic primary production could not meet the demand for particulate organic nitrogen of the cultured assemblages (oysters and epibiota). This emphasises the importance to the diet of cultured assemblages of other nitrogen sources such as heterotrophic populations of bacteria, cillates or flagellates.

In an attempt to develop a reproducible experimental model of bacterial infection in Crassostrea gigas, oysters taken from very localised sub-populations suffering natural mortality outbreaks were used in cohabitation trials under laboratory conditions. From these trials, a collection of Vibrio strains was isolated from moribund and healthy oysters. In a second step, strains were experimentally tested for virulence by means of injection into healthy oysters. This screening revealed a span of virulence among isolated strains from none to medium. When pooling injected strains, results suggest increased virulence. Vibrio strains may have additive/synergistic action leading to higher C, gigas mortality rates in experimental challenges. Although the study initially aimed to develop a simple experimental model, a complex of interactions emerged between several bacterial strains during the pathogenic process in their molluscan host. Selected strains provide a suitable model of experimental disease for further studies and better understanding of bacterial interaction and pathogenesis in C. gigas.

Apoptosis, or programmed cell death, has been reported as being pivotal in infectious diseases of different organisms. The effects of apoptosis on the progression and transmission of the protistan parasites Perkinsus marinus and Haplosporidium nelsoni in the eastern oyster Crassostrea virginica were studied. Oysters were diagnosed for their respective infections by standard methods, and apoptosis was detected using in situ hybridization to detect DNA fragments by end labeling on paraffin sections. A digoxigenin nucleotide probe was used to label the 200 bp fragment produced by apoptosis and detected immunohistochemically using an antidigoxigenin peroxidase conjugate. The probe/DNA fragment complex was stained with a peroxidase substrate and tissues were counterstained with methyl green. Uninfected oysters had large numbers of apoptotic hemocytes present in the connective tissue underlying the stomach, gill, and mantle epithelia, whereas oysters infected with P. marinus had a reduced number of apoptotic hemocytes. The parasite may prevent hemocyte apoptosis in order to yield a greater number of hemocytes in which to house itself. Large numbers of P. marinus cells in some infected oysters were eliminated via apoptosis in the stomach epithelia, disabling the spread of infectious particles through seawater. The oysters infected with H. nelsoni also had reduced numbers of apoptotic hemocytes, while part of the vesicular connective tissue cells were apoptotic. H. nelsoni plasmodia were eliminated via apoptosis in some oysters. Apoptosis may enhance progression and prevent transmission of infectious oyster diseases.

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.

Infections of a population of Crassostrea gigas by the copepod Mytilicola orientalis were examined at an oyster growing site at Dungarvan, County Waterford, Ireland. Twenty-one samples, each consisting of 20 to 30 oysters have been examined over 2 yr. Condition, sex, reproductive stage, length, weight, glycogen content and other parasite burdens of the oysters were examined in relation to the degree of infection of M. orientalis; 14.38% of oysters were infested. Mean abundance was 0.6 oyster -1 . The maximum number of copepods in an oyster was 20. M. orientalis had no effect on condition, growth, sex, stage or glycogen content of the oyster but correlated with shell burrowing by Polydora sp.

Mass mortalities of the Japanese pearl oyster Pinctada fucata martensii have widely occurred in western Japan since 1994. The causes of these mass mortalities are at present not thoroughly understood. In this study, we investigated oyster survival in relation to some environmental factors such as water temperature, concentration of chlorophyll a and density or composition of phytoplankton. The examined mass mortality occurred from September to December 1998, and the color on the adductor muscle of the oysters was red-brown, suggesting an infectious disease. Oysters that became moribund during the experiment lost weight, while the weight of unaffected oysters increased. The cell density of Nitzschia spp., an inedible algae for the oyster, in Uchiumi Bay increased before and during the mass mortality event. From the results of our study, we hypothesize that P. fucata martensii was weakened by starvation because of the dominance of inedible food and then contracted an infectious disease that resulted in mortality.

Concomitant sporadic high mortalities were reported in France in May 1994 among batches of hatchery-reared larval Pacific oysters Crassostrea gigas and European flat oysters Ostrea edulis in 2 hatcheries, and in June and July 1994 among batches of cultured spat of both species in a shellfish nursery. Histological observation showed the presence of cellular abnormalities in moribund animals. Transmission electron microscopy revealed the presence of herpes-like virus particles in infected larvae and spat of both oyster species. This is the first description of a herpes-like virus infection in larval O.. edulis. Viruses observed in diseased larvae and spat of both species are similar with respect to ultrastructure and morphogenesis, They were detected simultaneously in C. gigas and O. edulis larvae and spat, indicating possible interspecific transmission. Moreover, these viruses are associated with high mortality rates in both oyster species. An electron microscopic examination revealed hemocytes with condensed chromatin and extensive perinuclear fragmentation of chromatin. These data suggest that herpes-like viruses infecting oysters may induce apoptosis in oyster hemocytes.

In French Atlantic coastal ponds of Charente, oysters can grow under conditions where phytoplankton production is limited by nutrients exhaustion. Such ponds typically show a high concentration of ciliates and flagellates during the growing season (1 x 104 to 3 x 105 cells l-1 in June 1997). In order to evaluate the importance of the "protozoan trophic link " for energy transfer from the " microbial food web" to large benthic suspension feeders, we offered a coastal pond community of ciliates and flagellates as potential prey to the oyster, Crassostrea gigas. Clearance rate, filtered particles and relative retention efficiency were evaluated. In the grazing experiment, 94 % of ciliates and 86 % of flagellates (size between 4 and 72 μm), were retained by the oyster. Whatever their size, protists were similarly retained by the oyster gills. In terms of carbon, oyster retain on average 126 μg carbon (C) h-1 g-1 dry weight, a value over 4 times higher than reported for phytoplankton. These results indicate that a field community of protists can contribute in coastal oyster rearing ponds to the energy requirements of the oyster Crassostrea gigas. We report here the first experimental evidence of a significant retention of a protist community by oysters, supporting the role of protists as a trophic link between picoplankton and benthic filter-feeding bivalves.

During a detailed survey of Pacific oyster Crassostrea gigas juvenile health at intensive rearing facilities, an episode of persistent morbidity and mortality occurred over an 8 wk period in 1997. Affected oyster seed were typically between about 1.0 and 2.4 mm in shell height. Abscesses were formed in the extrapallial space resulting from invasion by straight bacterial rods along the inner shell surface. The abscesses contained host cells and bacteria. The infection appeared to be chronic, was associated with relatively low numbers of bacteria cells, and caused alterations of the underlying mantle and abnormal shell deposition. No signs of any other type of Infectious agent were found associated with the lesions. After a chronic time course, 2 outcomes were possible: either the mantle was breached, leading to an overwhelming terminal bacterial infection or, in some cases, host cell debris and bacteria were sequestered by new shell deposition and the infection was resolved. The condition can cause mortality and significant loss of growth in intensively cultured juvenile oysters.

Experiments in many parts of the world have indicated that sessile intertidal organisms are affected by fish predation. Farming of oysters Saccostrea commercialis (Iredale & Roughley) and Crassostrea gigas (Thunberg) in New South Wales, Australia, depends on their natural recruitment and growth on intertidal structures in estuaries. We investigated the effects of predation by fish on numbers of oysters recruiting to experimental panels of 3 different sizes. We tested effects of excluding fish of different sizes using different sizes of mesh (12.5 and 50 mm). Numbers of oysters were significantly reduced on panels open to predatory fish. Numbers of oysters per unit area and subsequent predation on them did not vary, however, across 3 different sizes of panels, indicating that predation was not dependent on patch size. Mortality on panels open to predation averaged 40.0% (±4.3% SE). Fish also significantly altered the distribution of sizes of oysters. The effect of predation was almost entirely attributable to toadfish Tetractenos spp. Previous knowledge of the life history of oysters and succession in these intertidal assemblages suggests that effects of predation may not, however, have important long-term consequences on natural populations.

Considering the economic importance of the Pacific oyster Crassostrea gigas to the French shellfish industry, the appearance of major diseases in this species could cause dramatic decreases in production. Suminoe oysters, of the non-indigenous species Crassostrea rivularis (Gould), were introduced into France to test their ability to adapt to local conditions. These oysters were imported after careful examination, and were maintained in laboratory quarantine. Some mortalities occurred 7 mo after importation. Histological and electron microscope examinations of 9 dead specimens revealed a parasite presumed to be a Bonamia-like protozoan. This is the first report concerning a parasite of the genus Bonamia in a species belonging to the genus Crassostrea. Thus, C. rivularis is not considered to be a suitable substitute for C. gigas in France.

This paper describes effects of temperature on herpes-like virus detection in Pacific oyster Crassostrea gigas larvae held at different temperatures. Intranuclear, intracytoplasmic and extracellular viral particles were observed in velum and mantle connective tissues of oyster larvae reared at 25-26°C. In larvae held at 22-23OC, although nuclear lesions were observed, the presence of viral particles was not detected. Results were obtained for oyster larvae with 4 different parental origins. Herpesvirus infection was found in 3 of the 4 groups of oyster larvae held In the same conditions at the higher temperature.

The susceptibility of the eastern oyster Crassostrea virginica to infection by the protozoan parasite Perlunsus mannus is Influenced by temperature Because of the crucial roles of heat shock proteins in cellular thermal tolerance and in host-parasite adaptations in other species we compared the in vitro heat shock responses of cultured P mannusand of oyster hemocytes The parasite and host heat shock proteins were different in size and in ~mmunochemical specificlty In addi t~on, the thermal threshold for induclng the iesponse was higher for P marinus acclimated to the same temperature as the oysters The results suggest that P marlnus is likely to employ heat shock proteins as part of ~ t s adaptive survival repertoire and that ~t nlay be able to function normally under conditions of hyperthermia that evoke an emergency physiological response from the oyster defense cells Furthermore, they indicate that ~t is feasible to Investigate the host's and parasite S adap t~ve responses to each other, since the11 individual responses are readily d~stinguishable

A low intensity Steinhausia-like infection was observed in h~stological sections of the gonads of 7 out of 10 female Sydney rock oysters Saccostrea comn~erc~alis obtained from a farm in Queensland. Australia. Parasitophorous vacuoles occurred within the nucleus and cytoplasm of ovocytes and contained up to 18 unicells (in cross section). Infection caused an Intense hemocyte infiltration. The parasite may cause problems In S. commercialis culture at hlgher infection intenslties.

On determine en laboratoire les taux de filtration, ingestion, assimilation, excretion et respiration d'O.c. en vue d'etablir le budget energetique en fonction de la taille et de deux concentrations de l'aliment (algue)

Nitrogen and oxygen fluxes were measured in situ during monthly dark enclosure experiments on oyster beds. The incubations were performed on undisturbed sediment with its endofauna, on 10 oysters Crassostrea gigas isolated from the substratum, and on 10 oysters associated with the sediment, to assess the impact of oyster farming on the environment. Ammonia was released by the sediment following a seasonal pattern, s ammonia excretion increased in spring (3.21 pm01 g-' h-') and autumn (2.5 to 6.7 pm01 g-' h-'), compared to low winter values (0.28 pm01 g-' h-'), in accordance with changes in temperature and oxygen consumption (0.14 and 1.3 mg O2 g-' h-' in winter and summer respectively). Some nitrate production was observed, suggesting occasional nitrification. Primary amine exchanges were rather erratic, whereas urea seems to play an important role seasonally as a nitrogenous end-product, in relation to a lower condition index at the end of the winter A sirnp!e budge? sf sediment-water exchanges, calculated on a m-' basis (e. g. 2 kg oysters m-'), indicates that over the year the oysters' contribution to the fluxes averages 37 and 40 % for ammonia and urea release respectively, and 26 O/ O for oxygen uptake. However, the actual contribution of oysters to the exchanges at the water-sediment interface rarely fits the expected values from potential estimates. Oxygen consumption, ammonia release and nitrate absorption of the association of oysters with sediment are mainly depressed, except durlng winter and early spring when respiration is stimulated.