Abstract: In recent years, the noninvasive monitoring of steroid hormone metabolites in feces of mammals and droppings of birds has become an increasingly popular technique. It offers several advantages and has been applied to a variety of species under various settings. However, using this technique to reliably assess an animal's adrenocortical activity is not that simple and straightforward to apply. Because clear differences regarding the metabolism and excretion of glucocorticoid metabolites (GCMs) exist, a careful validation for each species and sex investigated is obligatory. In this review, general analytical issues regarding sample storage, extraction procedures, and immunoassays are briefly discussed, but the main focus lies on experiments and recommendations addressing the validation of fecal GCM measurements in mammals and birds. The crucial importance of scrutinizing the physiological and biological validity of fecal GCM analyses in a given species is stressed. In particular, the relevance of the technique to detect biologically meaningful alterations in adrenocortical activity must be shown. Furthermore, significant effects of the animals' sex, the time of day, season, and different life history stages are discussed, bringing about the necessity to seriously consider possible sex differences as well as diurnal and seasonal variations. Thus, comprehensive information on the animals' biology and stress physiology should be carefully taken into account. Together with an extensive physiological and biological validation, this will ensure that the measurement of fecal GCMs can be used as a powerful tool to assess adrenocortical activity in diverse investigations on laboratory, companion, farm, zoo, and wild animals.

Abstract: The continuous development of tourism and related leisure activities is exerting an increasingly intense pressure on wildlife. In this study, a novel noninvasive method for measuring stress in the black grouse, an endangered, emblematic species of European ecosystems that is currently declining in several parts of its European range, is tested and physiologically validated. A radiometabolism study and an ACTH challenge test were performed on four captive black grouse (two of each sex) in order to get basic information about the metabolism and excretion of corticosterone and to find an appropriate enzyme-immunoassay (EIA) to measure its metabolites in the feces. Peak radioactivity in the droppings was detected within 1 to 2 hours. Injected (3)H-corticosterone was excreted as polar metabolites and by itself was almost absent. A cortisone-EIA was chosen from among seven tested EIAs for different groups of glucocorticoid metabolites, because it cross-reacted with some of the formed metabolites and best reflected the increase of excreted corticosterone metabolites, after the ACTH challenge test. Concentrations of the metabolites from fecal samples collected from snow burrows of free-ranging black grouse were within the same range as in captive birds. The noninvasive method described may be appropriate for evaluating the stress faced by free-living black grouse populations in the wild, particularly in mountain ecosystems where human disturbance, especially by winter sports, is of increasing conservation concern.

Abstract: Fecal steroid analyses are becoming increasingly popular among both field and laboratory scientists. The benefits associated with sampling procedures that do not require restraint, anesthesia, and blood collection include less risk to subject and investigator, as well as the potential to obtain endocrine profiles that are not influenced by the sampling procedure itself. In the feces, a species-specific pattern of metabolites is present, because glucocorticoids are extensively metabolized. Therefore, selection of adequate extraction procedures and immunoassays for measuring the relevant metabolites is a serious issue. In this review, emphasis is placed on the establishment and analytical validation of methods to measure glucocorticoid metabolites for a noninvasive evaluation of adrenocortical activity in droppings of birds.

Abstract: Neurobiological and behavioral research indicates that place learning and response learning occur simultaneously, in parallel. Such findings seem to conflict with theories of associative learning in which different cues compete for learning. The authors conducted place+response training on a radial maze and then tested place learning and response learning separately by reconfiguring the maze in various ways. Consistent with the effects of manipulating place and response systems in the brain (M. G. Packard & J. L. McGaugh, 1996), well-trained rats showed strong place learning and strong response learning. Three experiments using associative blocking paradigms indicated that prior response learning interferes with place learning. Blocking and related tests can be used to better understand how memory systems interact during learning.

Abstract: REASONS FOR PERFORMING STUDy: MP3 recorders are readily available, small, lightweight and low cost, providing the potential for logging analogue hoof mounted accelerometer signals for the characterisation of equine locomotion. These, however, require testing in practice. OBJECTIVES: To test whether 1) multiple MP3 recorders can maintain synchronisation, giving the ability to synchronise independent recorders for the logging of multiple limbs simultaneously; and 2) features of a foot mounted accelerometer signal attributable to foot-on and foot-off can be accurately identified from horse foot mounted accelerometers logged directly into an MP3 recorder. METHODS: Three experiments were performed: 1) Maintenance of synchronisation was assessed by counting the number of samples recorded by each of 4 MP3 recorders while mounted on a trotting horse and over 2 consecutive 30 min periods in 8 recorders on a bench. 2) Foot-on and foot-off times obtained from manual transcription of MP3 logged data and directly logged accelerometer signal were compared. 3) MP3/accelerometer acquisition units were used to log accelerometer signals from racehorses during extended training sessions. RESULTS: Mean absolute error of synchronisation between MP3 recorders was 10 samples per million (compared to mean number of samples, range 1-32 samples per million). Error accumulation showed a linear correlation with time. Features attributable to foot on and foot off were equally identifiable from the MP3 recorded signal over a range of equine gaits. CONCLUSIONS: Multiple MP3 recorders can be synchronised and used as a relatively cheap, robust, reliable and accurate logging system when combined with an accelerometer and external battery for the specific application of the measurement of stride timing variables across the range of equine gaits during field locomotion. POTENTIAL RELEVANCE: Footfall timings can be used to identify intervals between the fore and hind contacts, the identification of diagonal advanced placement and to calculate stride timing variables (stance time, protraction time and stride time). These parameters are invaluable for the characterisation and assessment of equine locomotion.