The effects of different holding methods on heart rate (HR) changes in the green crab, Carcinus maenas (Linnaeus, 1758), were investigated. Green crabs were held in perforated plastic boxes (with or without a layer of sand) suspended above the bottom of the tank or strapped to a weighted plastic grate. The HR of green crabs classified as unrestrained (plastic box with or without sand) dropped more rapidly compared with restrained (hanging from band, strapped to grate) green crabs. Within 1 h, unrestrained green crabs exhibited periods of cardiac pausing accounting for between 8% and 14% of the hourly time. In contrast, restrained green crabs rarely exhibited cardiac pausing. When the green crabs were subjected to a temperature increase (10–30 °C), the HR of unrestrained green crabs reached higher levels than that of the restrained animals. The four restraining methods were also used to investigate cardiac responses to hypoxia. During progressive hypoxia (100%–20% oxygen), the HR of unrestrained green crabs declined to lower levels than that of the restrained animals. The restraining methods appeared to be more stressful for the green crabs that maintained elevated HRs and were less able to respond to environmental change compared with green crabs that could move freely within a small chamber. This suggests that even subtle changes in experimental design may alter physiological responses.
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