Occurrence of OsHV-1 in Crassostrea gigas Cultured in Ireland during an Exceptionally Warm Summer. Selection of Less Susceptible Oysters.

The occurrence of OsHV-1, a herpes virus causing mass mortality in the Pacific oyster Crassostrea gigas was investigated with the aim to select individuals with different susceptibility to the infection. Naïve spat transferred to infected areas and juveniles currently being grown at those sites were analyzed using molecular and histology approaches. The survey period distinguishes itself by very warm temperatures reaching up to 3.5°C above the average. The virus was not detected in the virus free area although a spread of the disease could be expected due to high temperatures. Overall mortality, prevalence of infection and viral load was higher in spat confirming the higher susceptibility in early life stages. OsHV-1 and oyster mortality were detected in naïve spat after 15 days of cohabitation with infected animals. Although, infection was associated with mortality in spat, the high seawater temperatures could also be the direct cause of mortality at the warmest site. One stock of juveniles suffered an event of abnormal mortality that was significantly associated with OsHV-1 infection. Those animals were infected with a previously undescribed microvariant whereas the other stocks were infected with OsHV-1 µVar. Cell lesions due to the infection were observed by histology and true infections were corroborated by in situ hybridization. Survivors from the natural outbreak were exposed to OsHV-1 µVar by intramuscular injection and were compared to naïve animals. The survival rate in previously exposed animals was significantly higher than in naïve oysters. Results derived from this study allowed the selection of animals that might possess interesting characteristics for future analysis on OsHV-1 resistance.

Neutral red retention time assay in determination of toxicity of nanoparticles.

The neutral red retention time (NRRT) assay is useful for detecting decreased lysosomal membrane stability in haemocytes sampled from bivalves, a phenomenon often associated with exposure to environmental pollutants including nanomaterials. Bivalves are popular sentinel species in ecotoxicology and use of NRRT in study of species in the genus Mytilus is widespread in environmental monitoring. The NRRT assay has been used as an in vivo test for toxicity of carbon nanoparticles (Moore MN, Readman JAJ, Readman JW, Lowe DM, Frickers PE, Beesley A. 2009. Lysosomal cytotoxicity of carbon nanoparticles in cells of the molluscan immune system: An in vivo study. Nanotoxicology. 3 (1), 40-45). We here report application of this assay adapted to a microtitre plate format to a panel of metal and metal oxide nanoparticles (2 ppm). This showed that copper, chromium and cobalt nanoparticles are toxic by this criterion while gold and titanium nanoparticles are not. As the former three nanoparticles are often reported to be cytotoxic while the latter two are thought to be non-cytotoxic, these data support use of NRRT as a general in vitro assay in nanotoxicology.

A preliminary health survey of the hermit crab, Pagurus bernhardus, on the southwest coast of Ireland.

During October and November of 2011 and 2012, 229 hermit crabs, Pagurus bernhardus, were randomly collected from the intertidal shore at three locations along the southwest coast of Ireland. This is the first survey to assess the health status of this crustacean in Ireland. Carapace length and the sex of each crab were recorded. Crabs were screened for parasites by histology and the intensity and prevalence of infection was determined. Crabs of varying carapace length were screened. The only parasite observed in the crabs was Microphallus sp., the first record of this digenetic trematode in P. bernhardus in Europe.

Toxicity of copper oxide nanoparticles in the blue mussel, Mytilus edulis: a redox proteomic investigation.

Relatively little is known about the fate and effects of nanomaterials even in relatively simple organisms such as Mytilus edulis. Here, copper oxide nanoparticles (CuO NP) are shown to induce dose-dependent toxic effects at the biochemical, physiological and tissue levels in the blue mussel. Stable CuO NP suspensions were sized by differential light scattering and nanoparticle tracking analysis to yield average particle diameters of approximately 100 nm. These were administered to M. edulis, at doses of 400, 700 and 1000 ppb. Ingested copper was predominantly located in the gill tissue with small amounts in digestive gland. Fifteen coomassie-stained spots were excised from two dimensional gel electrophoresis separations of gill tissue extacts and identified by peptide mass fingerprinting. These contained six unique proteins (alpha- and beta-tubulin, actin, tropomyosin, triosephosphate isomerase and Cu-Zn superoxide dismutase). Of these, two spots (actin and triosephosphate isomerase) showed decreased protein thiols while three (alpha-tubulin, tropomyosin and Cu-Zn superoxide dismutase) showed increased carbonylation which is indicative of protein oxidation of cytoskeleton and enzymes in response to CuO NP. The neutral red retention time (NRRT) assay revealed toxicity due to the CuO NPs which was comparable with toxic metal oxide nanoparticles such as chromium and cobalt. In contrast, non-toxic titanium and gold metal oxide nanoparticles gave no NRRT effects at similar NP concentrations. Histology revealed deposition of pigmented brown cells in response to CuO NP, located predominantly along the mantle and gill margin but also lining digestive tubules and some of the sinuses and distributed throughout the connective tissue and in the adductor muscle.