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: Predation on livestock is a source of human-wildlife conflicts and can undermine the conservation of large carnivores. To design effective mitigation strategies, it is important to understand the determinants of predation across livestock species, which often differ in husbandry practices, vulnerability to predators and economic value. Moreover, attention should be given to both predation occurrence and intensity, because these can have different spatial patterns and predictors. We used spatial risk modelling to quantify factors affecting wolf predation on five livestock species in Portugal. Within the 1619 parishes encompassing the entire wolf range in the country, the national wolf compensation scheme recorded 17,670 predation events in 2009-2015, each involving one or more livestock species: sheep (31.7%), cattle (27.7%), goats (26.8%), horses (14.8%) and donkeys (3.2%). Models built with 2009-2013 data and validated with 2014-2015 data, showed a shared general pattern of predation probability on each species increasing with its own density and proximity to wolf packs. For some species there were positive relations with the density of other livestock species, and with habitat variables such as altitude, and land cover by shrubland and natural pastures. There was also a general pattern for predation intensity on each species increasing with its own density, while proximity to wolf packs had no significant effects. Predation intensity on goats, cattle and horses increased with the use of communal versus private pastures. Our results suggest that although predation may occur wherever wolves coexist with livestock species, high predation intensity is mainly restricted to particular areas where husbandry practices increase the vulnerability of animals, and this is where mitigation efforts should concentrate.

Abstract: We recorded the behaviour of dogs in detour tests, in which an object (a favourite toy) or food was placed behind a V-shaped fence. Dogs were able to master this task; however, they did it more easily when they started from within the fence with the object placed outside it. Repeated detours starting from within the fence did not help the dogs to obtain the object more quickly if in a subsequent trial they started outside the fence with the object placed inside it. While six trials were not enough for the dogs to show significant improvement on their own in detouring the fence from outside, demonstration of this action by humans significantly improved the dogs' performance within two-three trials. Owners and strangers were equally effective as demonstrators. Our experiments show that dogs are able to rely on information provided by human action when confronted with a new task. While they did not copy the exact path of the human demonstrator, they easily adopted the detour behaviour shown by humans to reach their goal.

Abstract: Innovative behaviors such as exploiting novel food sources can grant significant fitness benefits for animals, yet little is known about the mechanisms driving such phenomena, and the role of physiology is virtually unexplored in wild species. Two hypotheses predict opposing effects of physiological state on innovation success. On one hand, poor physiological condition may promote innovations by forcing individuals with poor competitive abilities to invent alternative solutions. On the other hand, superior physiological condition may ensure greater cognitive capacity and thereby better problem-solving and learning performance. To test these hypotheses, we studied the behavior of wild-caught house sparrows (Passer domesticus) in 4 novel tasks of food acquisition, one of which was presented to the birds in repeated trials, and we investigated the relationships of individual performance with relevant physiological traits. We found that problem-solving performance across the 4 tasks was moderately consistent within individuals. Birds with lower integrated levels of corticosterone, the main avian stress hormone, solved the most difficult task faster and were more efficient learners in the repeated task than birds with higher corticosterone levels. Birds with higher concentration of total glutathione, a key antioxidant, solved 2 relatively easy tasks faster, whereas birds with fewer coccidian parasites tended to solve the difficult task more quickly. Our results, thus, indicate that aspects of physiological state influence problem-solving performance in a context-dependent manner, and these effects on problem-solving capacity, probably including cognitive abilities, are more likely to drive individual innovation success than necessity due to poor condition.