Abstract: We investigated the impact of enclosure size and animal density on behavior and adrenocortical secretion in Pere David's deer in Dafeng Nature Reserve, China. From February 15 to April 16 in 2004, we conducted two experiments. First, we studied maintenance behavior and conflict behavior of Pere David's deer stags in a large enclosure (200 ha) with low animal density (0.66 deer/ha) and a small display pen (0.75 ha) with high animal density (25.33 deer/ha). The maintenance behavior we recorded included standing, locomotion, foraging and rest. During the behavioral observations, we collected fresh voided fecal samples from the stags periodically, and analyzed the fecal cortisol concentrations in those samples using radioimmunoassay technique. Second, we monitored the fecal cortisol concentrations of one group of stags (12 deer lived in an enclosure of 100 ha) before and after transferred into a small pen (0.5 ha). We found that in the first experiment: (1) there were significant differences in standing and rest whereas no significant differences of locomotion and foraging between the free-ranging group and the display group; (2) frequency of conflict behavior in the display group was significantly higher than those in the free-ranging group; and (3) fecal cortisol concentration of the display group (326.17+/-16.98 ng/g dry feces) was significantly higher than that of the free-ranging group (268.98+/-15.21 ng/g dry feces). In the second experiment, there was no significant difference of the fecal cortisol concentrations among sampling days, but the mean fecal cortisol concentration of the day after transferring (337.46+/-17.88 ng/g dry feces) was significantly higher than that of the day before transferring (248.44+/-7.99 ng/g dry feces). Comparison with published findings, our results indicated that enclosure size and animal density affect not only behaviors, but also adrenocortical secretion in Pere David's deer. Small living space with high animal density may impose physiological stress to captive Pere David's deer. Moreover, long-term physiological stress and increase of conflict behavior may subsequently affect survival and reproduction of the deer.

Abstract: During the past 20 years, measuring steroid hormone metabolites in fecal samples has become a widely appreciated technique, because it has proved to be a powerful, noninvasive tool that provides important information about an animal's endocrine status (adrenocortical activity and reproductive status). However, although sampling is relatively easy to perform and free of feedback, a careful consideration of various factors is necessary to achieve proper results that lead to sound conclusions. This article aims to provide guidelines for an adequate application of these techniques. It is meant as a checklist that addresses the main topics of concern, such as sample collection and storage, time delay extraction procedures, assay selection and validation, biological relevance, and some confounding factors. These issues are discussed briefly here and in more detail in other recent articles.

Abstract: Behavioural reactions to selected handling procedures were compared between conditioned, or imprint-trained, and untrained foals raised on the same farm. Nineteen randomly chosen healthy foals were imprint trained at birth and 24 h later (Group A). Twenty-one similar foals that were not imprint-trained served as age-matched controls (Group B). Training began within 10 min of birth and consisted of touch desensitization by gentle rubbing. Each tactile stimulus was repeated 30-50 times over 45-60 min, until the foal no longer resisted the procedure and appeared relaxed. The procedure was then repeated at 24 h of age. At that time a physical examination and blood analysis were performed to assess the foals' health status. Group B animals were not handled except for a brief physical examination and blood analysis at 24 h of age. Thereafter all foals were kept on pastures with their dams with no further handling until they were three months of age. Any foals handled for other reasons before that time were excluded from the study. At three months, each of the 28 foals that completed the study experienced the following handling procedures: acceptance of restraint, haltering, complete physical examination, acceptance of a plastic rebreathing bag, touching the whole body, intramuscular vaccination in the neck, intranasal vaccination, and deworming with oral paste. Response to each procedure was scored (1=not resistant, 2=low resistance, 3=strong resistance, 4=not possible without major physical restraint). Conditioned foals (Group A) were significantly less resistant to touching the front and hind legs and picking up the hind feet (P<0.05). The administration of vaccines and paste dewormer and the collection of blood were tolerated by the majority of the foals of both groups with no or low resistance. It appeared that neonatal imprint training resulted in a learned behaviour that resulted in decreased self-defence responses towards handling the limbs at three months of age.

Abstract: We examined whether very young dairy calves are able to discriminate between two people, and whether they use the colour of clothing or other indices to do so. During the familiarisation phase, one person (the familiar rewarder), who always wore the same colour clothes, gave milk, spoke gently and patted the calves in their individual pen for 6 days each week. During the test phase, the calf had to make a choice in an Y-maze placed in front of the gate of its stall. When the calves chose the familiar rewarder, they received 200 ml of milk as reinforcement. When they made the incorrect choice, they received nothing and were returned to their stall. On each test day, the calves made eight choices. The criterion of success was that the calf made at least six correct choices in eight trials on each of two consecutive test days (P<0.021 by the binomial law). The first experiment was carried out with fourteen 1-week-old male and female Holstein calves to see if calves could approach a person, who changed position in the maze, in order to obtain a feed reward. The familiar rewarder wore the same clothes as during the period of familiarisation and was in one arm of the Y-maze. The other arm was empty and the position of the familiar rewarder in the maze was randomised. Eleven of the 14 calves reached the criterion for success, after only three tests. The second experiment, carried out with five 2-week-old calves, examined whether the calves can differentiate the familiar rewarder (wearing the same clothing as during the period of familiarisation) from another person (the non-rewarder) wearing clothes of a different colour. The criterion of success was reached by all five calves. The third experiment was carried out with seven 2-week-old calves. It examined whether the calves can differentiate the familiar rewarder and the non-rewarder, when the two people are wearing clothes of the same colour (i.e. the same colour worn by the familiar rewarder during the phase of familiarisation). None of the calves were able to reach the criterion of success within a limited number of four test days. Often, calves would always choose the same arm of the maze. The fourth experiment was carried out on six 1-month-old calves. It was similar to experiment 3 with the difference that the familiar rewarder and the non-rewarder both wore the same colour clothes, but which were not the same colour as worn during the phase of familiarisation. Only one calf achieved the criterion of success within two test days. Results demonstrated that colour cues help very young calves to discriminate between two people, when these people wear different colour clothing. Some calves may be able to use other indicators than the colour of clothing. The Y-maze method is an promising way of examining calves' abilities to recognise people.

Abstract: This experiment investigated the effects of positive and negative tactile handling on the stress physiology and behaviour of dairy heifers. Forty-eight 5-14-month-old nonlactating Holstein-Friesian heifers were allocated to one of two handling treatments, either positive or negative tactile handling, over four time replicates. Handling was imposed twice daily, 2-5 min per session and involved moving animals individually along a 64 m outdoor route. The negatively handled heifers took longer to approach within 1 and 2 m of a stimulus person in a standard test, than their positively handled counterparts (P<0.001) and had a greater flight distance to an approaching stimulus (P<0.001). The time taken by the heifers to approach within 1 and 2 m of a familiar person was similar to that taken to approach within 1 and 2 m of an unfamiliar person in the standard test (P<0.05). There was a tendency for heifers to have a greater flight distance from the approaching unfamiliar person than from the approaching familiar person (P=0.06). The negatively handled heifers had greater (P<0.05) increases in total cortisol concentrations 5, 10 and 15 min after exposure to a human and had higher (P<0.05) free cortisol concentrations in the afternoon than the positively handled heifers. It is concluded that the nature of the human contact affects the subsequent behavioural response of heifers to humans. This behavioural response may extend to other humans through the process of stimulus generalisation, although there was some evidence of moderate discrimination. Negative handling results in an acute stress response in the presence of humans and also leads to a chronic stress response. Further research into the effect of these stress responses on milk production and welfare in fearful cows in a commercial situation is suggested.