Abstract: During the evolution of the horse, an extended period of feed intake, spread over the entire 24-h period, determined the horses� behaviour and physiology. Horses will not interrupt their feed intake for more than 4 h, if they have a choice. The aim of the present study was to investigate in what way restrictive feeding practices (non ad libitum) affect the horses� natural feed intake behaviour. We observed the feed intake behaviour of 104 horses on edible (n = 30) and non-edible bedding (n = 74) on ten different farms. We assessed the duration of the forced nocturnal feed intake interruption of horses housed on shavings when no additional roughage was available. Furthermore, we comparatively examined the feed intake behaviour of horses housed on edible versus non-edible bedding. The daily restrictive feeding of roughage (2 times a day: n = 8; 3 times a day: n = 2), as it is common in individual housing systems, resulted in a nocturnal feed intake interruption of more than 4 hours for the majority (74.32%, 55/74) of the horses on shavings (8:50 ± 1:25 h, median: 8:45 h, minimum: 6:45 h, maximum: 13:23 h). In comparison to horses on straw, horses on shavings paused their feed intake less frequently and at a later latency. Furthermore, they spent less time on consuming the evening meal than horses on straw. Our results of the comparison of the feed-intake behaviour of horses on edible and non-edible bedding show that the horses� ethological feeding needs are not satisfied on non-edible bedding. If the horses accelerate their feed intake (also defined as �rebound effect�), this might indicate that the horses� welfare is compromised. We conclude that in addition to the body condition score, the longest duration of feed intake interruption (usually in the night) is an important welfare indicator of horses that have limited access to roughage.

Abstract: Since the non-invasive field endocrinology techniques were developed, several fecal preservation and extraction methods have been established for a variety of species. However, direct adaptation of methods from previous studies for use in crested macaques should be taken with caution. We conducted an experiment to assess the accuracy and stability of fecal estrogen metabolite (E1C) and glucocorticoid metabolite (GCM) concentrations in response to several preservation parameters: (1) time lag between sample collection and fecal preservation; (2) long-term storage of fecal samples in 80% methanol (MeOH) at ambient temperature; (3) different degrees of feces drying temperature using a conventional oven; and (4) different fecal preservation techniques (i.e., freeze-drying, oven-drying, and field-friendly extraction method) and extraction solvents (methanol, ethanol, and commercial alcohol). The study used fecal samples collected from crested macaques (Macaca nigra) living in the Tangkoko Reserve, North Sulawesi, Indonesia. Samples were assayed using validated E1C and GCM enzyme immunoassays. Concentrations of E1C and GCM in unprocessed feces stored at ambient temperature remained stable for up to 8 h of storage after which concentrations of both E1C and GCM changed significantly compared to controls extracted at time 0. Long-term storage in 80% MeOH at ambient temperature affected hormone concentrations significantly with concentrations of both E1C and GCM increasing after 6 and 4 months of storage, respectively. Drying fecal samples using a conventional oven at 50, 70, and 90 °C did not affect the E1C concentrations, but led to a significant decline for GCM concentrations in samples dried at 90 °C. Different fecal preservation techniques and extraction solvents provided similar results for both E1C and GCM concentrations. Our results confirm previous studies that prior to application of fecal hormone analysis in a new species, several preservation parameters should be evaluated for their effects on hormone metabolite stability. The results also provide several options for fecal preservation, extraction, and storage methods that can be selected depending on the condition of the field site and laboratory.

Abstract: Although housing horses in single stalls limits their natural behavior to a great extent, this housing system is widespread in Germany, especially for competition horses. To improve the welfare of this system, free exercise on pastures or paddocks is deemed suitable, but it is also feared because of injuries and decreased willingness or motivation to perform. In the present study, three treatments were investigated with regard to their effect on the behavior of six competition horses in the stable, behavior during training, and on their degree of stress: daily training without free exercise (no turnout [NT]), solitary turnout for 2 hours after training, and 2-hour turnout in groups of two after training (group turnout). The horses' behavior in the stable was continuously analyzed through video recordings (2 pm to 6 am) on 3 days at the end of each treatment. The degree of stress was evaluated daily by heart rate variability at rest. The behavior during training was evaluated by a questionnaire answered by the riders, and the distance covered during training was measured by global positioning system. When NT was allowed, the horses showed less lying in the stable compared with the treatments with turnout. Heart rate variability measurements resulted in great individual differences, but generally, there was a higher degree of stress shown with the treatment NT according to the following parameters: standard deviation of inter-beat-intervals (SDNN), square root of the mean of the sum of the squares of differences between successive inter-beat-intervals (RMSSD), and ratio between low frequency and high frequency (LF/HF). The willingness to perform was evaluated as being slightly better in the treatments with turnout than in the treatment without turnout.