High-frequency recordings of valve opening behavior (VOB) in bivalves are often used to detect changes in environmental conditions. However, generally a single variable such as temperature or the presence of toxicants in the water is the focus. A description of routine VOB under non-stressful conditions is also important for interpreting responses to environmental changes. Here we present the first detailed quantitative investigation of the in-situ VOB of eastern oysters (Crassostrea virginica) to environmental variables typically not considered stressful. The VOB of eight individuals was monitored for seven weeks in a Louisiana estuary. We examined the relationships between VOB metrics (variance in mean % max opening among oysters, the probability of an oyster being closed, and the rate of valve closure), and temperature, salinity, chlorophyll-a (chl-a) concentration, the rate of change in those environmental variables, and the rate of change in water depth. Relationships were analyzed through statistical models including rates of change over 0, 0.25, 1-, 6-, 12-, and 24-hours. All the responses were best explained by the 12-hour time step model. The interaction effect between salinity and the rate of change of salinity had the greatest impact on variance in oysters’ behavior. Oysters closed faster at higher salinities and were more likely to be closed at lower chl-a concentrations. Significant interactions were found between many environmental variables, indicating a high level of complexity of oyster behavior in the natural environment. This study contributes to a better understanding of the impact of environmental conditions on oyster behavior and can help inform predictive tools for restoration initiatives and fisheries practices.