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.

Colour Preferences in Relation to Diet in Chimpanzees(Pan troglodytes), Gorillas (Gorilla gorilla) and Mandrills (Mandrillus sphinx).

Although trichromatic colour vision has been extensively studied as it grants significant advantages for Old World primates, it is unknown which selective pressures were behind the trait's evolution. The leading hypothesis would be that colour vision arose as a foraging adaptation because it allowed individuals to spot food more efficiently. To test this, we exposed 3 chimpanzees (Pan troglodytes), 5 gorillas (Gorilla gorilla) and 3 mandrills (Mandrillus sphinx) to colour cardboard plates to assess whether colours related to diet were the most preferred. The experimental setting was divided into two phases. During the first, animals were provided with colour cardboard plates of only 1 colour per data collection session. The order of colour presentation was randomly determined: white, black, yellow, green and red. In phase 2, primates were simultaneously provided with cardboard plates of all colours. Behavioural interactions with plates were measured using a one-zero group focal sampling (10-s sampling intervals and 20-min observation periods). Results showed that when animals were exposed to only 1 colour at a time, they exhibited different colour preferences depending on the species considered. Chimpanzees preferred red and yellow, the colours linked to fruits, while gorillas selected red and white. Mandrills exhibited fewer differences between colour preferences, with red being the most selected. Furthermore, when all colours were simultaneously provided, individuals chose colours related to diet over black and white. Although there were clear individual differences, our results support that trichromatic colour vision is an advantage in detecting and selecting red items. In the wild, it could be important in the detection of reddish fruits and leaves.

Testosterone and the dark ventral patch of male red deer: the role of the social environment.

The expression of male sexual traits, which is stimulated by testosterone, entails significant costs for individuals. Consequently, natural selection is expected to favour the modulation of sexual trait development according to the balance between its costs and benefits. The proportion of rivals in a population may affect this balance by increasing or decreasing the reproductive benefits associated with the development of sex traits. Here, we explore the relationship between testosterone level and sex trait size under two populational conditions of mate competition: fenced (i.e. high male-male competition; all male age groups are present) and unfenced (i.e. low competition; most males present are juveniles). Our model species is the Iberian red deer (Cervus elaphus hispanicus), and the sex trait is the dark ventral patch that males exhibit during the rutting season. Our results showed that the positive relationship between testosterone levels and the size of the dark ventral patch depends on the environmental level of male-male competition. Only in populations where the operational sex ratio was high (i.e. high proportion of rival males), individuals with high levels of testosterone developed the sex trait. Conversely, when mate competition was low, there was no significant relationship between testosterone level and trait size. This result reinforces the idea that the effect of testosterone in promoting the development of sex traits may be mediated by the intensity of mate competition in the population, as well as the role of sexual selection in the evolution of the dark ventral patch in red deer.

Reproductive hormones monthly variation in free-ranging European wildcats: Lack of association with faecal marking.

Integrating ecophysiological and behavioural discoveries in conservation and management plans is essential to preserve scarce and elusive species such as the European wildcat (Felis silvestris). The purpose of this study was to characterize the monthly variation in the steroid reproductive hormone metabolite levels (oestradiol, progesterone and testosterone) in this species and to test its possible association with a monthly pattern of faecal marking. By collecting fresh faecal samples in Montes do Invernadeiro Natural Park (Galicia, Northwest Spain) each month, we obtained a total of 110 samples belonging to 25 different individuals. We conducted enzyme immunoassays which allowed us to track the annual variation in reproductive hormone excretion patterns in wildcat scats. Furthermore, we also evaluated the possible relation between the faeces used as marks and the reproductive hormone levels. We found that oestradiol and progesterone metabolite levels exhibited a distinct pattern, both increasing during the breeding months. Oestradiol metabolite larger peaks were found during March and April, whereas the highest concentration of progesterone metabolites appeared in July. On the contrary, testosterone metabolite levels did not significantly change depending on the month. Moreover, we did not find any evidence that the faecal marking behaviour pattern was associated with reproductive hormone metabolite levels. It seems that other factors related to habitat and food resources could be more important in the performance of this behaviour.

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.

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.

A new large-scale index (AcED) for assessing traffic noise disturbance on wildlife: stress response in a roe deer (Capreolus capreolus) population.

Anthropogenic noise is a growing ubiquitous and pervasive pollutant as well as a recognised stressor that spreads throughout natural ecosystems. However, there is still an urgent need for the assessment of noise impact on natural ecosystems. This article presents a multidisciplinary study which made it possible to isolate noise due to road traffic to evaluate it as a major driver of detrimental effects on wildlife populations. A new indicator has been defined: AcED (the acoustic escape distance) and faecal cortisol metabolites (FCM) were extracted from roe deer faecal samples as a validated indicator of physiological stress in animals moving around in two low-traffic roads that cross a National Park in Spain. Two key findings turned out to be relevant in this study: (i) road identity (i.e. road type defined by traffic volume and average speed) and AcED were the variables that best explained the FCM values observed in roe deer, and (ii) FCM concentration was positively related to increasing traffic volume (road type) and AcED values. Our results suggest that FCM analysis and noise mapping have shown themselves to be useful tools in multidisciplinary approaches and environmental monitoring. Furthermore, our findings aroused the suspicion that low-traffic roads (< 1000 vehicles per day) could be capable of causing higher habitat degradation than has been deemed until now.

Changes in behaviour and faecal glucocorticoid levels in response to increased human activities during weekends in the pin-tailed sandgrouse.

Human recreational activities are becoming increasingly widespread and frequent, a fact that may potentially exacerbate their effects on wildlife. These human-related disturbances on animals may induce behavioural and physiological changes that can ultimately affect their fitness, showing a similar anti-predator response that against natural predator or other threats. Here, we combine the use of behavioural and physiological approaches to assess the potential effect of winter human activities on a threatened farmland bird in Europe, the pin-tailed sandgrouse (Pterocles alchata). We compared before, during and after weekend variations in human activity rates, pin-tailed sandgrouse behaviour (flocking and flying behaviour, interspecific association in mixed flocks and habitat use) and faecal glucocorticoid metabolite concentrations. Human disturbances, in particular those associated with hunting activities, peaked during weekends. Sandgrouse showed significant behavioural changes (increased sandgrouse-only flock sizes, increased proportion of birds flying and changes in habitat use) during weekends and higher faecal glucocorticoid metabolite concentrations after the weekends compared with during or before weekends. Therefore, physiological stress levels could be modulated by behavioural adjustments such as increased flock sizes and changes in habitat use that may allow sandgrouse to cope with increased human disturbance rates during weekends. Nevertheless, temporal and spatial organization of hunting days among groups of estates might be good strategies to buffer these potential adverse effects on wintering pin-tailed sandgrouse and other steppe species of conservation concern, while preserving a socio-economically important activity such as hunting.

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.

Onto the sense of smell in macaws, amazons and toucans: can they use volatile cues of fruits to make foraging decisions?

Over the past decades, empirical evidence has been accumulated indicating that olfactory information plays a fundamental role in bird life history. Nonetheless, many aspects of avian olfaction remain poorly understood. Our purpose was to broaden the knowledge about the importance of the sense of smell in some neglected bird groups: psittaciformes and ramphastids, and to compare how the response varied between the species. Because of the lack of information about the use of chemical cues for locating food in fruit-eating species, we also aimed to delve into this question. We conducted a 3-choice (water/vinegar/papaya and banana juice) scent test in 5 Costa Rican native species: scarlet macaw (Ara macao), red-lored amazon (Amazona autumnalis), yellow-naped amazon (Amazona auropalliata), keel-billed toucan (Ramphastos sulfuratus), and yellow-throated toucan (Ramphastos ambiguus). Results revealed that macaws and toucans allocated significantly more time to interacting with the fruit scent container, indicating that these species can perceive the volatile chemicals emitted by ripe fruits and that they can use this information to make foraging decisions. However, amazons did not dedicate more time to interact with the fruit treatment. Our research provides the first evidence of the ability to exploit chemical volatile cues in macaws and toucans.

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.

Reaction to Novel Objects and Fecal Glucocorticoid Metabolite Levels in Two Species of Nocturnal Geckos.

Many reptiles are maintained in captivity and heavily traded, although welfare measures for many species are not well established and are under-researched compared to other animals. In this study, we focused on two of these species: crested geckos (Correlophus ciliatus) and leopard geckos (Eublepharis macularius). To better interpret their behavior in captivity, the individual reaction to novel objects and the fecal glucocorticoid metabolite levels were measured in an attempt to identify the potential correlation between them. Also, we explored if some characteristic of the objects (e.g., color, shape, or smell) resulted in being more attractive to some species and/or individuals. Equivalent responses to different objects were not obtained for all the geckos, the behavioral response being highly individual and context-dependent, although modulated by the species. Individuals which manipulated earlier and interacted longer with novel objects showed lower basal fecal corticosterone metabolite (FCM) levels. Differences according to the species suggested that crested geckos have significantly greater and more variable FCM levels than leopard geckos. Our results can help to understand the reaction of geckos to novelty and have the potential to serve in their welfare assessment, although more studies are needed to proper establish welfare protocols.

Evaluating the functional, sexual and seasonal variation in the chemical constituents from feces of adult Iberian wolves (Canis lupus signatus).

Chemical signals deposited in feces play an important role in intraspecific and interspecific communication of many mammals. We collected fresh feces of adult wolves from wild breeding groups. All samples visually identified as belonging to wolves were subsequently identified to species level by sequencing a small fragment of mtDNA and sexed typing DBX6 and DBY7 sex markers. Using gas chromatography-mass spectrometry (GC-MS), we identified 56 lipophilic compounds in the feces, mainly heterocyclic aromatic organic compounds, such as indole or phenol, but also steroids, such as cholesterol, carboxylic acids and their esters between n-C4 and n-C18, aldehydes, alcohols and significant quantities of squalene and α-tocopherol, which would increase the chemical stability of feces on humid substrates. There was variability in the number and proportions of compounds between sexes, which could be indicative of their function as chemical signals. We also found variability in different reproductive states, especially in odorous compounds, steroids and α-tocopherol. Feces with a presumed marking function had higher proportions of α-tocopherol and steroids than feces with non-marking function. These compounds could be involved in intragroup and intergroup communication of wolves and their levels in feces could be directly related with the wolf's sex and physiological and reproductive status.

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.

The plant physical features selected by wildcats as signal posts: an economic approach to fecal marking.

The chemical signals of solitary and territorial felid species are essential for their intraspecific communication. We studied the selection of plant substrates during the fecal marking behavior of the European wildcat Felis silvestris from September 2008 to June 2009 in a protected area in Northwest Spain. The aim of the study was to examine the selection of plants as signal posts with respect to their physical characteristics. We hypothesized that wildcats deposit their fecal marks on plants with physical characteristics (e.g., size, species, and visual conspicuousness) that enhance the olfactory and visual effectiveness of the signal. Our results indicate that diameter, plant group, visual conspicuousness, and interaction between the diameter and plant group influence the decision of wildcats to deposit their fecal marks on plants. The wildcats chose plants with greater diameters and greater visual conspicuousness as scent-marking posts. Moreover, the wildcats chose woody and herbaceous plants, and certain plant species were marked more frequently than expected at random. Indeed, our results indicate that the fecal marks were not randomly distributed on the plants: the wildcats chose to place their marks on plants with certain physical characteristics that maximized the detectability of the signal by intruders and potential mates, thus facilitating the spatial distribution of the species.

Relationships between soil pollution by heavy metals and melanin-dependent coloration of a fossorial amphisbaenian reptile.

Melanin is the basis of coloration in many animals, and although it is often used in communication, thermoregulation, or camouflage, melanin has many other physiological functions. For example, in polluted habitats, melanin can have a detoxifying function. Melanic coloration would help to sequester in the skin the heavy metal contaminants from inside the body, which will be expelled to the exterior when the skin is sloughed. Moreover, animals should have evolved more melanic colorations in more polluted habitats ("industrial melanism" hypothesis). We examined whether the fossorial amphisbaenian reptile, Trogonophis wiegmanni, is able to eliminate heavy metals, derived from soil pollution by seagull depositions, through sloughing its skin. Our results suggest a covariation between levels of soil pollution by heavy metals and the concentration of heavy metals in the sloughed skins of amphisbaenians. This suggests that amphisbaenians may expel heavy metals from their bodies when they slough the skins. We also tested whether amphisbaenians inhabiting soils with higher levels of heavy metal pollution had darker (melanin-dependent) body colorations. However, contrary to predictions from the "industrial melanization" hypothesis, we found a negative relationship between soil pollution and proportions of melanic coloration. This contradictory result could, however, be explained because heavy metals have endocrine disruption effects that increase physiological stress, and higher stress levels could result in decreased melanogenesis. We suggest that although amphisbaenians might have some detoxifying mechanism linked to melanin in the skin, this process might be negatively affected by stress and result ineffective under conditions of high soil pollution.

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.

Effect of intensity and duration of anthropic noises on European mink locomotor activity and fecal cortisol metabolite levels.

Human activities involving noise emission can affect wild animals. European mink was exposed to road noise and human voice playbacks to analyze how sound intensity level and duration of both noises altered the time that individuals were active and if their fecal cortisol metabolite (FCM) levels varied. A Hierarchical Analysis Cluster was performed to establish 2 mink groups with respect to both noise source type: short duration/low intensity (SL) and long duration/high intensity (LH). We performed general linear mixed models to evaluate the variation in locomotor activity duration (s) and FCM (nanogram per gram) levels, respectively. The results showed both road noise and human voices decreased locomotor activity duration in SL more sharply compared with LH, and human voices were the triggers that induced the most pronounced response to both exposure conditions. FCM (ng/g) levels increased in SL compared with LH during road noise while the opposite happened during human voices. Differences based on sex and age of individuals were observed. In conclusion, noise characteristics given by the sound type determined the variations in locomotor activity duration while noise exposure level determined the variations in FCM (ng/g) levels. Attention should be paid to noisy activities (e.g., recreational activities for visitors in protected natural areas) and loud groups of people to conserve wildlife, especially noise sensitive species.

Hidden but Potentially Stressed: A Non-Invasive Technique to Quantify Fecal Glucocorticoid Levels in a Fossorial Amphisbaenian Reptile.

To understand wildlife responses to the changing environment, it is useful to examine their physiological responses and particularly their endocrine status. Here, we validated an enzyme immunoassay (EIA) to non-invasively quantify fecal corticosterone metabolites (FCM) in the fossorial amphisbaenian reptile Trogonophis wiegmanni from North Africa. We supplemented animals assigned to the treatment group with corticosterone dissolved in oil applied non-invasively on the skin for several days, while control groups received the oil-alone solution. Fresh feces were collected at the end of the supplementation period, and FCM levels were quantified by an EIA. Basal FCM levels were similar for both treatments and increased at the end of the test, but FCM increased significantly more in corticosterone-treated animals. A further examination of FCM levels in a wild population of this amphisbaenian did not find overall sexual, size or seasonal differences but showed a high range of variation among individuals. This suggests that different uncontrolled intrinsic or local environmental variables might increase the circulating glucocorticoid levels of different individuals. Our results confirmed the suitability of EIA for analyzing physiological changes in FCM in this amphisbaenian species. This technique may be useful for understanding and remediating the little-explored potential stressors of the soil environment that may negatively affect the health state of fossorial reptiles.