Metabolic Regulation of Copper Toxicity during Marine Mussel Embryogenesis.

The development of new tools for assessing the health of cultured shellfish larvae is crucial for aquaculture industries to develop and refine hatchery methodologies. We established a large-volume ecotoxicology/health stressor trial, exposing mussel (Perna canaliculus) embryos to copper in the presence of ethylenediaminetetraacetic acid (EDTA). GC/MS-based metabolomics was applied to identify potential biomarkers for monitoring embryonic/larval health and to characterise mechanisms of metal toxicity. Cellular viability, developmental abnormalities, larval behaviour, mortality, and a targeted analysis of proteins involved in the regulation of reactive oxygen species were simultaneously evaluated to provide a complementary framework for interpretative purposes and authenticate the metabolomics data. Trace metal analysis and speciation modelling verified EDTA as an effective copper chelator. Toxicity thresholds for P. canaliculus were low, with 10% developmental abnormalities in D-stage larvae being recorded upon exposure to 1.10 µg·L-1 bioavailable copper for 66 h. Sublethal levels of bioavailable copper (0.04 and 1.10 µg·L-1) caused coordinated fluctuations in metabolite profiles, which were dependent on development stage, treatment level, and exposure duration. Larvae appeared to successfully employ various mechanisms involving the biosynthesis of antioxidants and a restructuring of energy-related metabolism to alleviate the toxic effects of copper on cells and developing tissues. These results suggest that regulation of trace metal-induced toxicity is tightly linked with metabolism during the early ontogenic development of marine mussels. Lethal-level bioavailable copper (50.3 µg·L-1) caused severe metabolic dysregulation after 3 h of exposure, which worsened with time, substantially delayed embryonic development, induced critical oxidative damage, initiated the apoptotic pathway, and resulted in cell/organism death shortly after 18 h of exposure. Metabolite profiling is a useful approach to (1) assess the health status of marine invertebrate embryos and larvae, (2) detect early warning biomarkers for trace metal contamination, and (3) identify novel regulatory mechanisms of copper-induced toxicity.

Differential expression of novel metabolic and immunological biomarkers in oysters challenged with a virulent strain of OsHV-1.

Early lifestages of the Pacific oyster (Crassostrea gigas) are highly susceptible to infection by OsHV-1 µVar, but little information exists regarding metabolic or pathophysiological responses of larval hosts. Using a metabolomics approach, we identified a range of metabolic and immunological responses in oyster larvae exposed to OsHV-1 µVar; some of which have not previously been reported in molluscs. Multivariate analyses of entire metabolite profiles were able to separate infected from non-infected larvae. Correlation analysis revealed the presence of major perturbations in the underlying biochemical networks and secondary pathway analysis of functionally-related metabolites identified a number of prospective pathways differentially regulated in virus-exposed larvae. These results provide new insights into the pathogenic mechanisms of OsHV-1 infection in oyster larvae, which may be applied to develop disease mitigation strategies and/or as new phenotypic information for selective breeding programmes aiming to enhance viral resistance.

Establishing the thermal window for aerobic scope in New Zealand geoduck clams (Panopea zelandica).

Geoduck clams (Panopea spp.) are the longest-lived and largest deep burrowing bivalve. Their unique morphology allows them to live buried in the sediment at depths of up to 1 m. The endemic New Zealand geoduck (Panopea zelandica Quoy and Gaimard, 1835) has recently been identified as a potential species for aquaculture. However, very little is known about the biology and physiology of this entirely subtidal geoduck species. Currently, the New Zealand geoduck fishery relies entirely upon wild harvests, but farms are expected to emerge as cultivation protocols are established. A key step in the optimization of cultivation procedures is the identification of optimal temperature and food rations. One method for establishing thermal optima is to identify the temperature window that supports the widest aerobic scope: the degree to which metabolic rate can be increased to support elevated activity demands. Thus, we investigated the aerobic scope for activity at five different temperatures representative of typical environmental conditions (8, 11, 15, 19, and 23 °C) for juvenile and young adult P. zelandica. Clearance rate was also measured at all temperatures. Comparisons of aerobic scope for activity and clearance rates between size classes revealed that juvenile geoducks had a narrower thermal optimum than young adults (15-19 versus 11-19 °C, respectively). Temperatures higher than 19 °C resulted in a reduction of aerobic scope for activity and clearance rate for both juvenile and young adults, which may lead to reduced performance and elevated mortality. These findings provide the first measures of aerobic scope in P. zelandica, a key step towards a meaningful understanding of the ecophysiology of this unusual species.