Blind date: female fossorial amphisbaenians prefer scent marks of large and healthy males.

Selecting a good mate is a decision with important fitness consequences. For this reason, mate choice has promoted the evolution of sexual ornaments signaling the quality of an individual. In fossorial animals, inhabiting visually restricted underground environments, chemical senses should be very important for mate choice. We examined whether sexual chemical signals (substrate scent marks) produced by males of the Iberian worm lizard, Blanus cinereus, a strictly fossorial blind amphisbaenian, provide information to females on morphological traits and health state. We administered corticosterone (CORT) to males simulating a continuous stressor affecting their health. Females preferred settling at sites scent-marked by males in comparison with similar sites with female scent or unmarked sites, but the attractiveness of males' scent differed between individuals. Females preferred scent marks of larger/older males and with a higher immune response, while their body condition and CORT treatment were unrelated to female preferences. Chemical analyses showed that proportions of some compounds in precloacal secretions of males (used to produce scent marks) were correlated with the morphological (body size) and health state (immune response and body condition, but not CORT treatment) of these males. These results suggest that females may make site-selection decisions based on assessing the chemical characteristics of males' scent marks, which were reliably related to some of the traits of the male that produced the scent. Therefore, females might use chemical senses to increase the opportunities to find and mate with males of high quality, coping with the restrictions of the subterranean environment.

Wild Ungulates Constitute the Basis of the Diet of the Iberian Wolf in a Recently Recolonized Area: Wild Boar and Roe Deer as Key Species for Its Conservation.

The Iberian wolf (Canis lupus signatus) is recolonizing historical distribution areas after decades of absence. As in other human-dominated landscapes, finding a balance to protect this species by favoring recolonization and mitigating human-wildlife conflicts is a challenge. Since wolves are often generalist opportunistic predators, we studied their diet composition in central Spain to evaluate the consumption of domestic ungulates and provide reliable data that could help local authorities to deal with the current wolf-cattle ranchers conflict and coexistence. Diet composition (% prey occurrence, % prey ingested biomass) was analyzed through the identification of prey hairs present in 671 scats collected between 2017 and 2021. The wolves fed more on wild ungulates (82% occurrence) than domestic ones (18%). Wild boar (Sus scrofa, 44% occurrence) and roe deer (Capreolus capreolus, 35%) were the most consumed prey. The wolves positively selected these two species. The wolves' diets varied between seasons, years, and forest regions, but a diet based on wild ungulates predominated over domestic ones. Food niche breadth showed variations depending on seasons and years. Preserving the availability and diversity of wild ungulates may favor reducing livestock attacks and would be an achievable goal that would help to conserve this species and reduce conservation conflicts.

An Experimental Study in Wild Wood Mice Testing Elemental and Isotope Analysis in Faeces to Determine Variations in Food Intake Amount.

The analysis of carbon and nitrogen elemental (C, N) and isotopic compositions (δ13C, δ15N) in faeces are considered reliable methodologies for the study of diet in wildlife. Here, we tested the suitability of these techniques to detect variations in the amount of food intake. We captured wild wood mice (Apodemus sylvaticus) with Sherman live traps where bait access was initially free, and later it was experimentally limited inside by four different devices to cause intended variations in the amount ingested. The total C and N (%) and stable δ13C and δ15N isotopic values were determined for the bait and in mice faecal samples. Faecal values were lower than bait ones except for N, likely due to animal matter ingested before capture. No significant differences in total C, N and δ13C were found due to individual traits. However, breeding males showed higher δ15N values than breeding females, probably due to differences in energy and protein demands between both sexes during the breeding season. Only δ13C detected food intake variations (≥2 g). Despite further research being needed, these results initially support the potential of δ13C to provide information on the amount ingested, thus being useful to complement trophic ecology studies.

Ungulate presence and predation risks reduce acorn predation by mice in dehesas.

Foraging decisions by rodents are key for the long-term maintenance of oak populations in which avian seed dispersers are absent or inefficient. Decisions are determined by the environmental setting in which acorn-rodent encounters occur. In particular, seed value, competition and predation risks have been found to modify rodent foraging decisions in forest and human-modified habitats. Nonetheless, there is little information about their joint effects on rodent behavior, and hence, local acorn dispersal (or predation). In this work, we manipulate and model the mouse-oak interaction in a Spanish dehesa, an anthropogenic savanna system in which nearby areas can show contrasting levels of ungulate densities and antipredatory cover. First, we conducted a large-scale cafeteria field experiment, where we modified ungulate presence and predation risk, and followed mouse foraging decisions under contrasting levels of moonlight and acorn availability. Then, we estimated the net effects of competition and risk by means of a transition probability model that simulated mouse foraging decisions. Our results show that mice are able to adapt their foraging decisions to the environmental context, affecting initial fates of handled acorns. Under high predation risks mice foraged opportunistically carrying away large and small seeds, whereas under safe conditions large acorns tended to be predated in situ. In addition, in the presence of ungulates lack of antipredatory cover around trees reduced mice activity outside tree canopies, and hence, large acorns had a higher probability of survival. Overall, our results point out that inter-specific interactions preventing efficient foraging by scatter-hoarders can reduce acorn predation. This suggests that the maintenance of the full set of seed consumers as well as top predators in dehesas may be key for promoting local dispersal.

The dilution effect behind the scenes: testing the underlying assumptions of its mechanisms through quantifying the long-term dynamics and effects of a pathogen in multiple host species.

Disentangling the mechanisms that mediate the relationships between species diversity and disease risk has both theoretical and applied implications. We employed a model system of rodents and their Mycoplasma pathogens, in which an extreme negative diversity-disease relationship was demonstrated, to test the assumptions underlying three mechanisms that may explain this field pattern. Through quantifying the long-term dynamics and effects of the pathogen in its three host species, we estimated the between-host differences in pathogen spreading and transmission potentials, and host recovery potential and vulnerability to infection. The results suggest that one of the hosts is a pathogen amplifier and the other two hosts function as diluters. Considering the similarity in infection success and intensity among hosts, and the failure to detect any pathogen-induced damage, we could not validate the assumption underlying the hypotheses that diluters reduce the overall transmission or increase the mortality of infected hosts in the system. Instead, the results demonstrate that diluters clear the infection faster than amplifiers, supporting the possibility that the addition of diluters to the community may reduce the overall number of infected hosts through this mechanism. This study highlights the contribution of experimental studies that simultaneously explore different aspects of host-pathogen interactions in multiple hosts, in diversity-disease research.

Non-Invasive Monitoring of Adrenocortical Activity in Three Sympatric Desert Gerbil Species.

The study of the endocrine status can be useful to understand wildlife responses to the changing environment. Here, we validated an enzyme immunoassay (EIA) to non-invasively monitor adrenocortical activity by measuring fecal corticosterone metabolites (FCM) in three sympatric gerbil species (Gerbillus andersoni, G. gerbillus and G. pyramidum) from the Northwestern Negev Desert's sands (Israel). Animals included into treatment groups were injected with adrenocorticotropic hormone (ACTH) to stimulate adrenocortical activity, while control groups received a saline solution. Feces were collected at different intervals and FCM were quantified by an EIA. Basal FCM levels were similar in the three species. The ACTH effect was evidenced, but the time of FCM peak concentrations appearance differed between the species (6-24 h post-injection). Furthermore, FCM peak values were observed sooner in G. andersoni females than in males (6 h and 18 h post-injection, respectively). G. andersoni and G. gerbillus males in control groups also increased FCM levels (18 h and 48 h post-injection, respectively). Despite the small sample sizes, our results confirmed the EIA suitability for analyzing FCM in these species as a reliable indicator of the adrenocortical activity. This study also revealed that close species, and individuals within a species, can respond differently to the same stressor.

Losing seasonal patterns in a hibernating omnivore? Diet quality proxies and faecal cortisol metabolites in brown bears in areas with and without artificial feeding.

Bears are omnivores particularly well-adapted to variations in the nutritional composition, quality and availability of food resources. Artificial feeding practices have been shown to strongly influence diet composition and seasonality, as well as to cause alterations in wintering and movement in brown bears (Ursus arctos). In this study, we investigated seasonal differences (hypophagia vs hyperphagia) in food quality of two brown bear subpopulations in the Polish Carpathians using faecal nitrogen (FN) and carbon (FC) estimates. The subpopulations inhabit areas that differ in artificial feeding practices: no artificial feeding occurs in the western subpopulation (Tatra Mountains), while artificial food targeted to ungulates is provided and used year-round in the eastern subpopulation (Bieszczady Mountains). We also compared these results with faecal cortisol metabolites (FCM) to explore how FN and FC correlate with the hypothalamic-pituitary-adrenal axis activity and if the seasonal patterns are apparent. We found that in Tatra Mts bears fed on significantly higher quality diet, as shown by FN and FC values, and had significantly higher FC levels in hyperphagia, when they accumulate fat reserves for wintering. The pattern in FCM levels for Tatra subpopulation followed the changes in energy intake during the seasons of hypo- and hyperphagia, while in Bieszczady Mts, the area with intensive feeding, no seasonal patterns could be observed. Artificial feeding practices may disrupt nutrient phenology and seasonality, relative to subpopulations with natural diets. We showed that the availability of human-provided foods may alter not only the overall dietary quality, but also hormonal patterns linked to seasonal nutritional requirements. Combining FN, FC and FCM proved to be a useful tool for reconstructing diet quality and related physiological patterns.

The dynamics between limited-term and lifelong coinfecting bacterial parasites in wild rodent hosts.

Interactions between coinfecting parasites may take various forms, either direct or indirect, facilitative or competitive, and may be mediated by either bottom-up or top-down mechanisms. Although each form of interaction leads to different evolutionary and ecological outcomes, it is challenging to tease them apart throughout the infection period. To establish the first step towards a mechanistic understanding of the interactions between coinfecting limited-term bacterial parasites and lifelong bacterial parasites, we studied the coinfection of Bartonella sp. (limited-term) and Mycoplasma sp. (lifelong), which commonly co-occur in wild rodents. We infected Bartonella- and Mycoplasma-free rodents with each species, and simultaneously with both, and quantified the infection dynamics and host responses. Bartonella benefited from the interaction; its infection load decreased more slowly in coinfected rodents than in rodents infected with Bartonella alone. There were no indications for bottom-up effects, but coinfected rodents experienced various changes, depending on the infection stage, in their body mass, stress levels and activity pattern, which may further affect bacterial replication and transmission. Interestingly, the infection dynamics and changes in the average coinfected rodent traits were more similar to the chronic effects of Mycoplasma infection, whereas coinfection uniquely impaired the host's physiological and behavioral stability. These results suggest that parasites with distinct life history strategies may interact, and their interaction may be asymmetric, non-additive, multifaceted and dynamic through time. Because multiple, sometimes contrasting, forms of interactions are simultaneously at play and their relative importance alternates throughout the course of infection, the overall outcome may change under different ecological conditions.

The role of steroid hormones and individual traits in food intake in the wood mouse (Apodemus sylvaticus).

Availability of food resources affects animal survival and reproduction. Thus, coping with changes in food availability is one of the most crucial behavioural and physiological processes in wildlife. Food intake is a key concept in animal ecology that is directly conditioned by food quality and abundance or diet choice, but may also vary according to individual-related factors (e.g. foraging behaviours, social rank or energy-demanding periods) and the influence of the endocrine system on energy metabolism. Here, we studied food intake in relation to individual characteristics (sex, breeding condition and age) and whether steroid hormones (testosterone and corticosterone metabolites) mediate food intake in wild wood mouse (Apodemus sylvaticus). Field work was carried out in February-March 2014 in Monte de Valdelatas (Madrid, Spain). Wood mice were live-trapped for 10 consecutive days in four independent plots. Traps were baited with 4 g of toasted corn and food intake was calculated by subtracting the remaining bait found inside traps. Fresh faecal samples from 130 different wood mice were collected and faecal testosterone and corticosterone metabolites (FTM and FCM, respectively) were analysed by enzyme immunoassays. Food intake was higher in females than males, probably due to greater energy requirements. Non-breeders and young individuals also showed a higher food intake. These individuals usually hold a lower social rank which is associated to limited food resources because of dominants; thus, increased food intake may be a result of freely exploit food bait inside traps while avoiding risky competition. In addition, food intake negatively correlated with FTM levels and positively with FCM levels indicating that both hormones have an active role mediating food intake in the wood mouse. Our data suggest that food intake is a function of both individual traits and the endocrine system that accordingly respond throughout different energy-demanding periods.

Wood mouse feeding effort and decision-making when encountering a restricted unknown food source.

Animals making foraging decisions must balance the energy gained, the time invested, and the influence of key environmental factors. In our work, we examined the effect of predation risk cues and experience on feeding efforts when a novel food resource was made available. To achieve this, we live-trapped wood mouse Apodemus sylvaticus in Monte de Valdelatas (Madrid), where 80 Sherman traps were set in four plots. Traps were subjected to two food-access difficulties in treatments consisting of three consecutive nights: open plastic bottles (easy) and closed bottles (difficult), both using corn as bait. To simulate predation risk, we set fox faeces in half of the traps in each plot. We also considered moonlight (medium/low) as an indirect predation risk cue. We analysed whether bottles had been bitten by mice and the gnawed area of each bottle was measured. Our results indicated that food access difficulty, experience, and predation risk determined mice feeding decisions and efforts. The ability of mice to adapt feeding effort when a new food source is available was demonstrated because a higher proportion of closed bottles exhibited bite marks and the gnawed area was bigger. Moreover, mouse experience was determinant in the use of this new resource since recaptured mice gnawed broader orifices in the bottles and the gnawed area increased each time an individual was recaptured. Additionally, direct predation risk cues prompted mice to bite the bottles whereas the effect of different moon phases varied among the food access treatments. This study provides direct evidence of formidable efficacy of wild mice to exploit a new nutrient resource while considering crucial environmental factors that shape the decision-making procedure.

Stressful living in lower-quality habitats? Body mass, feeding behavior and physiological stress levels in wild wood mouse populations.

Wild populations are continuously subjected to changes in environmental factors that pose different challenges. Body condition and hormones have been commonly used as health indicators due to their potential correlation with fitness. In the present study, we analyzed whether habitats of different quality influenced body mass, food intake and physiological stress levels in wild wood mice (Apodemus sylvaticus). Field work was seasonally carried out in Holm oak woods and pine forests in central Spain. A total of 93 wood mice from 4 different populations (2 per habitat type) were live-trapped. From each captured individual we noted body mass and food intake, measured as the amount of bait remaining in each trap. The physiological stress levels were measured non-invasively in collected fresh feces by quantifying fecal corticosterone metabolites (FCM) with a 5a-pregnane-3ß,11ß, 21-triol-20-one enzyme immunoassay. Wood mice abundances decreased from spring to summer, were higher in Holm oak woods than in pine forests and also resulted in different age-class distribution between both habitats. Individuals inhabiting pine forests showed a lower body mass and increased food intake, probably because of the comparatively lower food quality and availability in this habitat. Furthermore, these individuals showed increased physiological stress levels, likely due to the lower quality habitat in relation to both food and vegetation cover availability. Overall, besides affecting local wood mouse abundance, our study underscores the effect of habitat quality on body mass, food intake and the endocrine stress response. Considering the wood mouse's pivotal position in ecosystems, these results could help in the understanding of environmental traits hampering the viability of wild populations.

Foraging, feeding, and physiological stress responses of wild wood mice to increased illumination and common genet cues.

In nature, animals are exposed to a broad range of threats imposed by predators, which may strongly influence the ecology of prey species directly or indirectly by affecting their behavior via fear of predation. Here, we studied wood mice Apodemus sylvaticus behavioral and physiological responses to simulated predation risk. Risk avoidance was analyzed by live trapping with control traps and traps treated with feces of common genet Genetta genetta (direct cue of risk) under new moon nights and following by simulated full moon conditions (indirect cue). The time devoted to foraging behavior and capture time were analyzed by video recording mice activity around traps. Food intake was calculated based on the amount of bait remaining in each trap. Fecal corticosterone metabolites (FCMs) were measured by enzyme-immunoassay as indicators of physiological stress responses. Fewer wood mice were captured during full moon, yet only non-breeding adult males clearly avoided common genet odor. Mice were captured sooner at night during the simulated full moon conditions and later in predator-treated traps. Foraging activity was lower when individuals faced predator's feces, but neither food intake nor FCM levels were affected by predation risk cues. Direct and indirect cues of predation risk selectively affected wood mice behavior, although behavioral responses seem to be modulated by different costs-benefit balances related to the individual's perception of risk. The lack of physiological responses to predation risk cues suggests that wood mice did not perceive them as reliable stressors or the response was too small or transient to be measured by FCM.

The concentration of fear: mice's behavioural and physiological stress responses to different degrees of predation risk.

Predation is an unavoidable and dangerous fact in the lifetime of prey animals and some sign of the proximity of a predator may be enough to trigger a response in the prey. We investigated whether different degrees of predation risk by red foxes (Vulpes vulpes) evoke behavioural and physiological stress responses in wood mice (Apodemus sylvaticus). We examined the variation in mice responses due to individual factors (sex and reproductive status) and related them to the concentration of the volatile compounds from fox faeces over time. In our experiment, we introduced predation cues into four plots, each subjected to a different concentration treatment (0, 10, 50 and 100% concentration of fresh faeces of red fox), based on the following outline: initial odourless phase 0, phase1 in which predation treatment was renewed daily, and phase 2 in which we renewed the treatment only on the first day. Wood mice were live trapped during all three phases and the physiological response was measured non-invasively by analysing faecal corticosterone metabolites (FCM) in freshly collected faeces. Data were analysed by Generalized Linear Mixed Models. Overall, males were trapped less often than females, and reproductively active individuals from both sexes avoided traps more than non-reproductively active individuals, especially in medium- and high- concentration plots. Variations in FCM concentrations were explained by plot, the interaction between plot and treatment phase, and the interaction between the treatment phase and the reproductive status. During phase 1, we detected a significant rise in FCM levels that increased with predator faecal odour concentration. Additionally, reproductively active individuals showed a strong physiological response during both phases 1 and 2 in all plots, except the control plot. Our results indicated that wood mice are able to discriminate different degrees of predation risk, which allows them to trigger gradual changes in their behavioural and physiological stress responses.

Are motorways potential stressors of roadside wood mice (Apodemus sylvaticus) populations?

Linear infrastructures represent one of the most important human impacts on natural habitats and exert several effects on mammal populations. Motorways are recognized as a major cause of habitat fragmentation and degradation and of biodiversity loss. However, it is unknown whether motorways lead to increased physiological stress reactions in wild animal populations. We analysed faecal corticosterone metabolites (FCM) in wild populations of wood mice (Apodemus sylvaticus) living in a well-preserved Mediterranean agro-pastoral woodland at different distances (verge, 500 m and 1000 m) from the AP-51 motorway in Spain. Wood mice were captured with Sherman live traps, and fresh faecal samples from 424 individuals were collected and analyzed in the laboratory. The quantification of FCM was performed by a 5α-pregnane-3ß,11ß, 21-triol-20-one enzyme immunoassay. Results showed that females had higher FCM levels than males, and these levels were higher in breeding females. In addition, FCM levels were positively correlated with body weight of individuals. Wood mice captured where cattle were present showed higher FCM levels than individuals living where cattle were not detected. FCM levels were higher in non-breeding individuals living close to the motorway compared with FCM levels in those individuals captured further from the motorway. This is the first study showing evidence of the motorways' impact on physiological stress reactions in wild wood mice populations. Understanding how free-living animals are influenced by human interventions could help to understand other subtle changes observed in wild animal populations. Since mice are used world-wide as research models these results could open new perspectives testing human influence on the natural environment and trade-offs of species in degraded ecosystems.