Instability of fecal glucocorticoid metabolites at 4°C: Time to freeze matters.

The use of fecal corticosteroid metabolites (FCMs) has proven to be well suited to evaluate adrenocortical activity, a major component of the stress response, particularly in wildlife. As with any tools, confounding factors and drawbacks must be carefully considered. Among them, sample preservation and storage are of particular importance, as they can affect stability of FCMs and lead to biased results and interpretations. Arguably, immediate freezing of fecal samples upon collection is the best practice to preserve FCM integrity, however, for logistical reasons, this condition is rarely feasible in the field. It is generally argued that temporary storage of samples at low above-zero temperature is an acceptable way of preserving samples in the field before freezing them for long-term storage. However, to our knowledge, there is no empirical study that demonstrates the stability of fecal metabolites in samples stored at +4°C. In this study, we collected a fresh fecal sample from 20 captive roe deer, each of which was homogenized and split into three subsamples (60 subsamples in total) to investigate the effects on FCMs levels of temporary storage at +4°C for 24 h and 48 h before freezing versus immediate freezing at -20°C after feces collection. Compared to immediate freezing, mean FCMs levels decreased by 25% every 24 h when feces were stored at +4°C before freezing. The variance of FCMs levels followed the same pattern, leading to a clear reduction in the ability to detect biological effects. Minimizing the storage time at +4°C before freezing should therefore be seriously considered when establishing sampling and storage protocols for feces in the field for adequate hormonal profiling.

Covariation between glucocorticoids, behaviour and immunity supports the pace-of-life syndrome hypothesis: an experimental approach.

The biomedical literature has consistently highlighted that long-term elevation of glucocorticoids might impair immune functions. However, patterns are less clear in wild animals. Here, we re-explored the stress-immunity relationship considering the potential effects of behavioural profiles. Thirteen captive roe deer (Capreolus capreolus) were monitored over an eight-week period encompassing two capture events. We assessed how changes in baseline faecal cortisol metabolite (FCM) concentrations following a standardized capture protocol and an immune challenge using anti-rabies vaccination affected changes in 13 immune parameters of innate and adaptive immunity, and whether these changes in baseline FCM levels and immune parameters related to behavioural profiles. We found that individuals with increased baseline FCM levels also exhibited increased immunity and were characterized by more reactive behavioural profiles (low activity levels, docility to manipulation and neophilia). Our results suggest that the immunity of large mammals may be influenced by glucocorticoids, but also behavioural profiles, as it is predicted by the pace-of-life syndrome hypothesis. Our results highlight the need to consider covariations between behaviour, immunity and glucocorticoids in order to improve our understanding of the among-individual variability in the stress-immunity relationships observed in wildlife, as they may be underpinned by different life-history strategies.

Ranging behaviour of roe deer in an experimental high-density population: are females territorial?

We studied the ranging behaviour and spatial relationships between seven roe deer during more than 4 years in a partly wooded 14.2-ha enclosure. The animals (three young males, four adult females) were monitored with GPS telemetry collars. As expected, the surface area and overlap of the males' bimonthly ranges decreased, and the distance between their arithmetic centres increased, as they became adult and, for two of them, territorial. Unexpectedly, females also tended to space out, the surface area and overlap of their bimonthly ranges being minimal in May to June, i.e. during the birth period. The distance between their arithmetic centres reached its maximum at the same time. Overlap between females' ranges was consistently lower than those between males and females' ranges, or between 1-year old males' ranges. Our results raise the questions of female seasonal territoriality and of independence of the spacing systems of the two sexes in roe deer.

Spatial organization and spatial distribution of activities within home ranges in a Springbok (Antidorcas marsupialis) captive population.

We studied over 1 year the spatial organization and the spatial distribution of activities in a captive springbok (Antidorcas marsupialis) population living in an 18-ha enclosure located in southern France. Throughout the study period, the two adult males occupied fairly exclusive home ranges, in the overlapping part of which the three subadult males were restricted. The spatial and temporal distribution of aggressive, marking, and avoidance behavior of males showed that the two adults were territorial, except during summer. They accounted for 71% of all marking behaviors recorded, for 77% of the aggressive behavior, and for 91% of the sexual interactions, whereas subadult males accounted for 94% of the avoidance behavior observed. The adult females used the whole enclosure, moving through the males' home ranges. They fed everywhere, but they all had the same preferred resting area, located in the center of the territory of one of the two adult males. They gave birth, accounted for maternal behavior and were engaged in sexual interactions in sectors differing from one individual to the other, but mainly outside the sector where all males' home ranges overlapped. Our results are compared to those reported in natural conditions and lead us to discuss both the functional interpretations of marking behavior, and the signification of a home range for an ungulate. Zoo Biol 27:19-35, 2008. (c) 2007 Wiley-Liss, Inc.

Herd size in large herbivores: encoded in the individual or emergent?

In large mammalian herbivores, the increase of group size with habitat openness was first assumed to be an adaptive response, encoded in the individual. However, it could, alternatively, be an emergent property: if groups were nonpermanent units, often fusing and splitting up, then any increase of the distance at which animals perceive one another could increase the rate of group fusion and thus mean group size. Dynamical models and empirical data support this second hypothesis. This is not to say that adaptive modifications of mean herd size cannot occur. However, this changes the way in which we can envisage the history of gregariousness in large herbivores during the Tertiary.