Abstract: Adult male elephants periodically show the phenomenon of musth, a condition associated with increased aggressiveness, restlessness, significant weight reduction and markedly elevated androgen levels. It has been suggested that musth-related behaviours are costly and that therefore musth may represent a form of physiological stress. In order to provide data on this largely unanswered question, the first aim of this study was to evaluate different assays for non-invasive assessment of adrenocortical function in the male African elephant by (i) characterizing the metabolism and excretion of [3H]cortisol (3H-C) and [14C]testosterone (14C-T) and (ii) using this information to evaluate the specificity of four antibodies for determination of excreted cortisol metabolites, particularly with respect to possible cross-reactions with androgen metabolites, and to assess their biological validity using an ACTH challenge test. Based on the methodology established, the second objective was to provide data on fecal cortisol metabolite concentrations in bulls during the musth and non-musth condition. 3H-C (1 mCi) and 14C-T (100 microCi) were injected simultaneously into a 16 year old male and all urine and feces collected for 30 and 86 h, respectively. The majority (82%) of cortisol metabolites was excreted into the urine, whereas testosterone metabolites were mainly (57%) excreted into the feces. Almost all radioactive metabolites recovered from urine were conjugated (86% 3H-C and 97% 14C-T). In contrast, 86% and >99% of the 3H-C and 14C-T metabolites recovered from feces consisted of unconjugated forms. HPLC separations indicated the presence of various metabolites of cortisol in both urine and feces, with cortisol being abundant in hydrolysed urine, but virtually absent in feces. Although all antibodies measured substantial amounts of immunoreactivity after HPLC separation of peak radioactive samples and detected an increase in glucocorticoid output following the ACTH challenge, only two (in feces against 3alpha,11-oxo-cortisol metabolites, measured by an 11-oxo-etiocholanolone-EIA and in urine against cortisol, measured by a cortisol-EIA) did not show substantial cross-reactivity with excreted 14C-T metabolites and could provide an acceptable degree of specificity for reliable assessment of glucocorticoid output from urine and feces. Based on these findings, concentrations of immunoreactive 3alpha,11-oxo-cortisol metabolites were determined in weekly fecal samples collected from four adult bulls over periods of 11-20 months to examine whether musth is associated with increased adrenal activity. Results showed that in each male levels of these cortisol metabolites were not elevated during periods of musth, suggesting that in the African elephant musth is generally not associated with marked elevations in glucocorticoid output. Given the complex nature of musth and the variety of factors that are likely to influence its manifestation, it is clear, however, that further studies, particularly on free-ranging animals, are needed before a possible relationship between musth and adrenal function can be resolved. This study also clearly illustrates the potential problems associated with cross-reacting metabolites of gonadal steroids in EIAs measuring glucocorticoid metabolites. This has to be taken into account when selecting assays and interpreting results of glucocorticoid metabolite analysis, not only for studies in the elephant but also in other species.

Abstract: This paper describes the results of a series of studies showing that variations in mother-pup interactions program the development of individual differences in behavioral and endocrine stress responses in the rat. These effects are associated with altered expression of genes in brain regions, such as the amygdala, hippocampus, and hypothalamus, that regulate the expression of stress responses. Studies from evolutionary biology suggest that such “maternal effects” are common and often associated with variations in the quality of the maternal environment. Together these findings suggest an epigenetic process whereby the experience of the mother alters the nature of the parent-offspring interactions and thus the phenotype of the offspring.

Abstract: The development of litter and individual differences in the rate of ultrasonic signaling of neonatal rats was studied. Systematic variations among litters and individuals emerged, without differential treatment. These differences were not correlated with variations in general development as indexed by body weight. Two experiments using a cross-fostering design showed that litter differences developed independently of variations in postnatal environment. These results indicate that the variations among litters in ultrasound rate have a prenatal, possibly genetic, etiology and may represent reliable indicants of response to environmental stress.

Abstract: Social stress is frequently used as a model for studying the neuroendocrine mechanisms underlying stress-induced behavioral inhibition, depression, and fear conditioning. It has previously been shown that social subordination may result in increased glucocorticoid release and changes in brain signaling systems. However, it is still an open question which neuroendocrine and behavioral differences are causes, and which are consequences of social status. Using juvenile rainbow trout of similar size and with no apparent differences in social history, we demonstrate that the ability to win fights for social dominance can be predicted from the duration of a behavioral response to stress, in this case appetite inhibition after transfer to a new environment. Moreover, stress responsiveness in terms of confinement-induced changes in plasma cortisol was negatively correlated to aggressive behavior. Fish that exhibited lower cortisol responses to a standardized confinement test were markedly more aggressive when being placed in a dominant social position later in the study. These findings support the view that distinct behavioral-physiological stress coping styles are present in teleost fish, and these coping characteristics influence both social rank and levels of aggression.

Abstract: For free-living or feral horses weaning takes place naturally at around 8-9 months [Gill, E.L., 1988. Factors affecting body condition of New Forest Ponies. Ph.D. Thesis. Department of Biology, University of Southampton]. Some mares will continue to suckle their foal until shortly before the arrival of their next foal, gestation being approximately 342 days depending upon the breed of the horse [Ropiha, R.T., Mathews, G., Butterfield, R.M., 1969. The duration of pregnancy in Thoroughbred mares. Vet. Rec. 84, 552-555]. Under domestic conditions, weaning tends to take place earlier, typically between 4 and 6 months of age. The weaning process has been identified as associated with potential psychological, physical and nutritional stressors that are of welfare concern. Following a review of the literature it is evident that there is a need for detailed research into what should constitute best practice with respect to foal and mare welfare. In addition, there is a need to understand the potential long-term impact of weaning on, for example, trainability and later maternal behaviour, and whether the stresses associated with early weaning have detrimental effects on the performance horse. There is also a lack of clear information concerning the most frequently observed weaning practices and the reasons why certain weaning methods are chosen. Some variables should be closely managed during weaning in order to minimise stress responses. These include: early creep feeding to familiarise the young animal with the food it will be exposed to during weaning, feeding a high fibre diet and keeping the animal in extensive conditions using a gradual approach to weaning. However, we conclude that there may not be one best method for weaning, since the chosen method must take into account a number of factors including: available resources, the housing environment, the individual foal's stage of development, the strength of the mare-foal attachment, the foal's ability to cope with changes in social conditions and the ability of the horse owner to implement the chosen method. We do however suggest that the fewest stress responses appear to occur where foals are weaned gradually and allowed to have social contact either with other foals or with older horses.