Modelling interactions of acid-base balance and respiratory status in the toxicity of metal mixtures in the American oyster Crassostrea virginica.

Heavy metals, such as copper, zinc and cadmium, represent some of the most common and serious pollutants in coastal estuaries. In the present study, we used a combination of linear and artificial neural network (ANN) modelling to detect and explore interactions among low-dose mixtures of these heavy metals and their impacts on fundamental physiological processes in tissues of the Eastern oyster, Crassostrea virginica. Animals were exposed to Cd (0.001-0.400 microM), Zn (0.001-3.059 microM) or Cu (0.002-0.787 microM), either alone or in combination for 1 to 27 days. We measured indicators of acid-base balance (hemolymph pH and total CO(2)), gas exchange (Po(2)), immunocompetence (total hemocyte counts, numbers of invasive bacteria), antioxidant status (glutathione, GSH), oxidative damage (lipid peroxidation; LPx), and metal accumulation in the gill and the hepatopancreas. Linear analysis showed that oxidative membrane damage from tissue accumulation of environmental metals was correlated with impaired acid-base balance in oysters. ANN analysis revealed interactions of metals with hemolymph acid-base chemistry in predicting oxidative damage that were not evident from linear analyses. These results highlight the usefulness of machine learning approaches, such as ANNs, for improving our ability to recognize and understand the effects of sub-acute exposure to contaminant mixtures.

Differential localization and bacteriostasis of Vibrio campbellii among tissues of the Eastern oyster, Crassostrea virginica.

In bivalve mollusks the roles of individual tissues in antimicrobial defense remain unclear. In this study, Crassostrea virginica were injected in the adductor muscle with 10(5) live Vibrio campbellii. Major tissues were dissected at 10, 30, 60 or 120 min postinjection (PI); in each tissue undegraded (intact) bacteria were quantified by real-time PCR and culturable bacteria were enumerated by selective plating. At 10 min PI, accumulation of bacteria varied among tissues from approximately 2.4 x 10(3) (labial palps, digestive gland) to 24.2 x 10(3) (gonads) intact Vibrio g(-1). Neither distribution nor accumulation of intact bacteria changed with time except in the hemolymph. In most tissues, more than 80% of intact bacteria were culturable at 10 min PI and culturability decreased with time. In contrast, only 19% of intact bacteria in gonadal tissue could be cultured at 10 min PI, pointing to a major role for the gonadal tissues in antibacterial defense of molluscs.