Abstract: Faecal steroid hormone metabolites are becoming increasingly popular as parameters for reproductive functions and stress. The extraction of the steroids from the faecal matrix represents the initial step before quantification can be performed. The steroid metabolites present in the faecal matrix are of varying polarity and composition, so selection of a proper extraction procedure is essential. There have been some studies to address this complex but often neglected point. Radiolabelled steroids (e.g. cortisol or progesterone) have frequently been added to faecal samples to estimate the efficiency of the extraction procedures used. However, native, unmetabolized steroids are normally not present in the faeces and therefore the results are artifi- cial and do not accurately reflect the actual recoveries of the substances of interest. In this respect, recovery experiments based on faecal samples from radiometabolism studies are more informative. In these samples, the metabolite content accurately reflects the mixture of metabolites present in the given species. As a result, it is possible to evaluate different extraction methods for use with faecal samples. We present studies on sheep, horses, pigs, hares and dogs that utilized samples containing naturally metabolized, 14C-labelled steroids. We recommend extracting faecal steroids by simply suspending the faeces in a high percentage of a primary alcohol (for glucocorticoid metabolites 80% aqueous methanol proved best suited for virtually all mammalian species tested so far). Not only does the procedure significantly increase the total amount of recovered radioactivity, it also increases the percentage of unconjugated metabolites, which are more likely to be recognized by the antibodies used in various immunoassays. The advantages of this extraction procedure are clear: it is very easy to use (no evaporation step is needed), it yields high recoveries and variation based on the extraction procedure is reduced to a minimum.

Abstract: Mortality is high in zoo-housed black rhinoceros (Diceros bicornis), and the reproductive rates of captive white rhinoceros (Ceratotherium simum) are unsustainably low. To determine the possible role of stress in the causation of these problems, we analyzed weekly fecal samples collected for 1 year from black (10 males and 16 females) and white (six males and 13 females) rhinoceroses at 16 zoos for corticoid metabolite concentrations. Fecal corticoid profiles were examined in relation to behavior as rated by keepers in a questionnaire, luteal phase ovarian cycles of females (Brown et al., 2001), and socioenvironmental factors. We compared individual fecal corticoid profiles by examining hormone means and variability (i.e., standard deviation (SD) and coefficient of variation (CV)). For the black rhinos, higher mean corticoid concentrations were found at zoos where rhinos were maintained in enclosures that were exposed to the public around a greater portion of the perimeter. Higher variability in corticoid excretion was correlated with higher rates of fighting between breeding partners and higher institutional mortality rates. Black rhino pairs that were kept separated exhibited lower corticoid variability and less fighting activity when they were introduced during female estrous periods compared to pairs that were kept together every day. For white rhinos, significantly lower mean corticoids were found for individuals that rated higher on “friendliness to keeper.” Higher corticoid variability was found in noncycling as compared to cycling white rhino females. Noncycling females exhibited higher rates of stereotypic pacing and lower frequencies of olfactory behaviors. Interindividual differences in mean corticoids in both species appeared to be related to responsiveness to humans, whereas corticoid variability was related to intraspecific social relationships. More importantly, high corticoid variability appeared to be an indicator of chronic or “bad” stress, because of its association with potentially deleterious consequences in each species (i.e., fighting and mortality (black rhino), and reproductive acyclicity (white rhino)). Our results provide evidence that social stressors may cause chronic stress in black and white rhinos, and that this contributes to the captive-population sustainability problems observed in each species. Zoo Biol 0:1–18, 2005. © 2005 Wiley-Liss, Inc.

Abstract: Background Recent record rainfall in much of semi-arid Central Australia is the most likely reason for a feral horse population increase in excess of normal. Uncontrolled numbers of feral horses have habitat degradation and animal welfare implications. Objectives The aims of this study were to investigate the social structure of feral horses and assess their population growth rate following unseasonably high rainfall. Methods The study area was 4000 km2 of unmanaged, semi-arid country in Central Australia (latitude 24.50°S, longitude 132.10°E). Horses were identified by descriptive features from ground searches, movement-activated cameras and ‘hides’ positioned at key water holes. Wherever possible, sex and age categories were documented. Population growth rate was estimated by the number of foals divided by the number of horses older than 1 year in the observed population. Results A total of 1424 horses were identified and categorised, of which 335 were foals born within the current year. Only 123 juveniles were identified. Of the adult horses, 53.4% were male and 46.6% were female and this differed from parity (P = 0.04). Of the mares, 71.9% had a foal at foot and the population growth rate was 29.5%. Conclusions With a sustained population growth rate of 29.5%, this population of feral horses will more than double within 3 years. The high population increase will likely have a detrimental effect on native fauna and flora and the fragile, semi-arid ecosystems of Central Australia. After a period of high rainfall and plentiful resources, ‘normal’ drought conditions will return and many feral horses will starve and die as they compete for limited resources.

Abstract: It is known that the right and left brain hemispheres differ in their ability to regulate autonomic processes in the organism. Direct unilateral stimulation of the brain provokes side-dependent endocrine, immune and other visceral reactions. Since brain hemispheres are mainly involved in the regulation of muscles and sensory organs on the contra lateral side of the body the activation of behavioural asymmetry stimulates the contra lateral half of the brain. The important theoretical and practical question of whether autonomic processes can be regulated via the behavioural asymmetry route remains unexplored. In this study, we report that the chronic presentation of an emotionally important stimulus--food--from the left side, improves reproductive performance in animals in a broad range of feeding conditions. The unilateral presentation of food can also influence lactation, but in this case the side-dependent effects are different under varying feeding conditions. This finding opens a simple practical approach to influence basic somatic functions in the organism.

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.