Non-invasive measurement of steroids in fish-holding water: important considerations when applying the procedure to behaviour studies

Fish behaviourists are increasingly turning to non-invasive measurement of steroid hormones in holding water, as opposed to blood plasma. When some of us met at a workshop in Faro, Portugal, in September, 2007, we realised that there were still many issues concerning the application of this procedure that needed resolution, including: Why do we measure release rates rather than just concentrations of steroids in the water? How does one interpret steroid release rates when dealing with fish of different sizes? What are the merits of measuring conjugated as well as free steroids in water? In the ‘static’ sampling procedure, where fish are placed in a separate container for a short period of time, does this affect steroid release—and, if so, how can it be minimised? After exposing a fish to a behavioural stimulus, when is the optimal time to sample? What is the minimum amount of validation when applying the procedure to a new species? The purpose of this review is to attempt to answer these questions and, in doing so, to emphasize that application of the non-invasive procedure requires more planning and validation than conventional plasma sampling. However, we consider that the rewards justify the extra effort.

Physical Health Problems and Environmental Challenges Influence Balancing Behaviour in Laying Hens

With rising public concern for animal welfare, many major food chains and restaurants are changing their policies, strictly buying their eggs from non-cage producers. However, with the additional space in these cage-free systems to perform natural behaviours and movements comes the risk of injury. We evaluated the ability to maintain balance in adult laying hens with health problems (footpad dermatitis, keel damage, poor wing feather cover; n = 15) using a series of environmental challenges and compared such abilities with those of healthy birds (n = 5). Environmental challenges consisted of visual and spatial constraints, created using a head mask, perch obstacles, and static and swaying perch states. We hypothesized that perch movement, environmental challenges, and diminished physical health would negatively impact perching performance demonstrated as balance (as measured by time spent on perch and by number of falls of the perch) and would require more exaggerated correctional movements.We measured perching stability whereby each bird underwent eight 30-second trials on a static and swaying perch: with and without disrupted vision (head mask), with and without space limitations (obstacles) and combinations thereof. Video recordings (600 Hz) and a three-axis accelerometer/gyroscope (100 Hz) were used to measure the number of jumps/falls, latencies to leave the perch, as well as magnitude and direction of both linear and rotational balance-correcting movements. Laying hens with and without physical health problems, in both challenged and unchallenged environments, managed to perch and remain off the ground. We attribute this capacity to our training of the birds. Environmental challenges and physical state had an effect on the use of accelerations and rotations to stabilize themselves on a perch. Birds with physical health problems performed a higher frequency of rotational corrections to keep the body centered over the perch, whereas, for both health categories, environmental challenges required more intense and variable movement corrections. Collectively, these results provide novel empirical support for the effectiveness of training, and highlight that overcrowding, visual constraints, and poor physical health all reduce perching performance.