Pacific oysters, Crassostrea gigas, living at a range of tidal heights, routinely encounter large seasonal fluctuations in temperature. We demonstrate that the thermal limits of oysters are relatively plastic, and that these limits are correlated with changes in the expression of one family of heat-shock proteins (HSP70). Oysters were cultured in the intertidal zone, at two tidal heights, and monitored for changes in expression of cognate (HSC) and inducible (HSP) heat-shock proteins during the progression from spring through winter. We found that the "control" levels (i.e., prior to laboratory heat shock) of HSC77 and HSC72 are positively correlated with increases in ambient temperature and were significantly higher in August than in January. The elevated level of HSCs during the summer was associated with moderate, 2-3 degrees C, increases in the upper thermal limits for survival. We measured concomitant increases in the threshold temperatures (T(on)) required for induction of HSP70. Total hsp70 mRNA expression reflected the seasonal changes in the expression of inducible but not cognate members of the HSP70 family of proteins. A potential cost of increased T(on) in the summer is that there was no extension of the upper thermal limits for survival (i.e., induction of thermotolerance) after sublethal heat shock at temperatures that were sufficient to induce thermotolerance during the winter months.

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This study investigated temperature effects on depuration for reducing Vibrio parahaemolyticus and Vibrio vulnificus in American oyster (Crassostrea virginica). Raw oysters were inoculated with 5-strain cocktail of V. parahaemolyticus or V. vulnificus to levels of 10(4) to 10(5) MPN (most probable number)/g and depurated in artificial seawater (ASW) at 22, 15, 10, and 5 degrees C. Depuration of oysters at 22 degrees C had limited effects on reducing V. parahaemolyticus or V. vulnificus in the oysters. Populations of V. parahaemolyticus and V. vulnificus were reduced by 1.2 and 2.0 log MPN/g, respectively, after 48 h of depuration at 22 degrees C. Decreasing water temperature to 15 degrees C increased the efficacy of depuration in reducing V. parahaemolyticus and V. vulnificus in oysters. Reductions of V. parahaemolyticus and V. vulnificus in oysters increased to 2.1 and 2.9 log MPN/g, respectively, after 48 h of depuration at 15 degrees C. However, depurations at 10 and 5 degrees C were less effective than at 15 degrees C in reducing the Vibrio spp. in oysters. Extended depuration at 15 degrees C for 96 h increased reductions of V. parahaemolyticus and V. vulnificus in oysters to 2.6 and 3.3 log MPN/g, respectively.

Summer seed mortality (SSM) has occurred yearly in Tomales Bay, California since 1993. SSM has resulted in up to 90% cumulative losses, and has been associated with extreme temperature, phytoplankton blooms, and an oyster herpesvirus. In this study, three stocks of Pacific oysters were planted at three sites in California (Inner Tomales Bay, Outer Tomales Bay, and Bodega Harbor) in October of 2000 (Fall) and April of 2001 (Spring) and monitored for mortality, growth, and health status. In April of 2001, a similar study was conducted in Totten Inlet, WA state using cohorts of oysters planted in California; animals were monitored for mortality and growth. Temperature data were collected at all sites; phytoplankton abundance data were collected at the California sites. Mortality occurred only at the Inner Tomales Bay site where losses were correlated with maximum temperatures (r ¼ 0.949) and preferentially affected faster growing oysters (r ¼ 0.916). Significant differences in cumulative mortality were identified among oysters stocks and two of the three oysters stocks planted in the fall outperformed their cohorts planted in the spring (P < 0.0001). Microscopic changes in connective tissue and digestive tubules are consistent with previous observations of herpesvirus infections in oysters including: diffuse to multifocal pertibular hemocyte infiltration, diapedesis, dilation of the digestive tubules, nuclear hypertrophy, and chromatin margination. Nuclear hypertrophy and chromatin margination, in particular, are suggestive of herpesvirus infections; these histological changes were rare indicating the need to use multiple diagnostic methods when oyster herpesviruses are suspected to cause SSM. Temperature maxima (;25°C) experienced at the Inner Tomales Bay site are not considered extreme for Pacific oyster survival; the association between oyster herpesviruses and temperature in Tomales Bay, California is discussed.

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