Contribution of host species and pathogen clade to snake fungal disease hotspots in Europe.

Infectious diseases are influenced by interactions between host and pathogen, and the number of infected hosts is rarely homogenous across the landscape. Areas with elevated pathogen prevalence can maintain a high force of infection and may indicate areas with disease impacts on host populations. However, isolating the ecological processes that result in increases in infection prevalence and intensity remains a challenge. Here we elucidate the contribution of pathogen clade and host species in disease hotspots caused by Ophidiomyces ophidiicola, the pathogen responsible for snake fungal disease, in 21 species of snakes infected with multiple pathogen strains across 10 countries in Europe. We found isolated areas of disease hotspots in a landscape where infections were otherwise low. O. ophidiicola clade had important effects on transmission, and areas with multiple pathogen clades had higher host infection prevalence. Snake species further influenced infection, with most positive detections coming from species within the Natrix genus. Our results suggest that both host and pathogen identity are essential components contributing to increased pathogen prevalence.

Ecological responses of squamate reptiles to nocturnal warming.

Nocturnal temperatures are increasing at a pace exceeding diurnal temperatures in most parts of the world. The role of warmer nocturnal temperatures in animal ecology has received scant attention and most studies focus on diurnal or daily descriptors of thermal environments' temporal trends. Yet, available evidence from plant and insect studies suggests that organisms can exhibit contrasting physiological responses to diurnal and nocturnal warming. Limiting studies to diurnal trends can thus result in incomplete and misleading interpretations of the ability of species to cope with global warming. Although they are expected to be impacted by warmer nocturnal temperatures, insufficient data are available regarding the night-time ecology of vertebrate ectotherms. Here, we illustrate the complex effects of nocturnal warming on squamate reptiles, a keystone group of vertebrate ectotherms. Our review includes discussion of diurnal and nocturnal ectotherms, but we mainly focus on diurnal species for which nocturnal warming affects a period dedicated to physiological recovery, and thus may perturb activity patterns and energy balance. We first summarise the physical consequences of nocturnal warming on habitats used by squamate reptiles. Second, we describe how such changes can alter the energy balance of diurnal species. We illustrate this with empirical data from the asp viper (Vipera aspis) and common wall lizard (Podarcis muralis), two diurnal species found throughout western Europe. Third, we make use of a mechanistic approach based on an energy-balance model to draw general conclusions about the effects of nocturnal temperatures. Fourth, we examine how warmer nights may affect squamates over their lifetime, with potential consequences on individual fitness and population dynamics. We review quantitative evidence for such lifetime effects using recent data derived from a range of studies on the European common lizard (Zootoca vivipara). Finally, we consider the broader eco-evolutionary ramifications of nocturnal warming and highlight several research questions that require future attention. Our work emphasises the importance of considering the joint influence of diurnal and nocturnal warming on the responses of vertebrate ectotherms to climate warming.

Pesticide concentrations in a threatened freshwater turtle (Emys orbicularis): Seasonal and annual variation in the Camargue wetland, France.

Wetlands are among the most threatened ecosystems on the planet and pollution is a major factor causing the decline of wetland biodiversity. Despite the increasing use of pesticides, their fate and effects on freshwater reptiles remain largely unknown. We studied the European pond turtle (Emys orbicularis), a long-lived species at risk with a high exposure potential to pesticides. Between 2018 and 2020, we measured 29 pesticides and metabolites in 408 blood samples of turtles from two populations in the Camargue wetland (France). We were able to quantify 24 compounds and at least one pesticide or one degradation product in 62.5% of samples. Pesticide occurrences and concentrations were low, except for a herbicide widely used in rice cultivation and locally detected in water: bentazone that reached high blood concentrations in E. orbicularis. The occurrence and the concentration of pesticides in E. orbicularis blood depended mainly on the site and the sampling date in relation to pesticide application. Individual characteristics (sex, age, body condition) did not explain the occurrence or the concentration of pesticides found in turtle blood. Assessing the exposure of aquatic wildlife to a cocktail of currently-used pesticides is a first and crucial step before studying their effects at the individual and population levels.

Prey consumption does not restore hydration state but mitigates the energetic costs of water deprivation in an insectivorous lizard.

To cope with limited availability of drinking water in their environment, terrestrial animals have developed numerous behavioral and physiological strategies including maintaining an optimal hydration state through dietary water intake. Recent studies performed in snakes, which are generalist carnivorous reptiles, suggest that the benefits of dietary water intake are negated by hydric costs of digestion. Most lizards are generalist insectivores that can shift their prey types, but firm experimental demonstration of dietary water intake is currently missing in these organisms. Here, we performed an experimental study in the common lizard Zootoca vivipara, a keystone mesopredator from temperate climates exhibiting a great diversity of prey in its mesic habitats, in order to investigate the effects of food consumption and prey type on physiological responses to water deprivation. Our results indicate that common lizards cannot improve their hydration state through prey consumption, irrespective of prey type, suggesting that they are primarily dependent upon drinking water. Yet, high-quality prey consumption reduced the energetic costs of water deprivation, potentially helping lizards to conserve a better body condition during periods of limited water availability. These findings have important implications for understanding the physiological responses of ectotherms to water stress, and highlight the complex interactions between hydration status, energy metabolism and feeding behavior in insectivorous lizards.

What drives the risk of being bitten by a viper? A fine spatial scale study in western France.

Previous studies on viper bites in France have focused on clinical consequences of envenomation, efficacy of antivenom and epidemiology of bites. Herein, we wanted to clarify temporal and spatial patterns in bite incidence using a fine spatial scale (municipality level). We focused on viper bites recorded over the last 10 years in 4 regions of western France. We addressed the determinants of bite occurrence and number of bites considering the following variables: predicted probability of viper presence, species (V. aspis or V. berus), climatic data, tourism function rate, soil transformation and landscape use. 703 bite cases were retained with significant disparities between areas. Bites occurred either during a garden-related activity (339 cases, 51.2%) or during an activity in the countryside (300 cases, 45.3%). The probability of presence of a viper at the municipality level positively influenced the risk of being bitten (multiplied by 3 for a variation in probability of 0.25 from 0.5) but varied between species (lower in V. berus than V. aspis). Artificial land development had a positive effect on bite risks. Finally, a tourism function rate above 50 beds/100 inhabitants was strongly associated with an increase in the risk of occurrence and frequency of bites. Overall, viper bites recorded in our study were concentrated on the south coastline of Pays de la Loire region. The coastal towns are significant areas of tourist attraction and are located close to preserved semi-natural landscapes that provide favorable habitats for vipers. This convergence may favor human/wildlife encounters.

Behavioural hydroregulation protects against acute effects of drought in a dry-skinned ectotherm.

During extreme climate events, behavioural thermoregulation may buffer ectotherms from thermal stress and overheating. However, heatwaves are also combined with dry spells and limited water availability, and how much individuals can behaviourally mitigate dehydration risks through microclimate selection remains largely unknown. Herein, we investigated the behavioural and physiological responses to changes in air and microhabitat humidity in a terrestrial ectotherm, the asp viper (Vipera aspis). We exposed individuals to a simulated heatwave together with water deprivation for 3 weeks, and manipulated air water vapour density (wet air vs. dry air) and microclimate (wet shelter vs. dry shelter) in a two-by-two factorial design. Dry air conditions led to substantial physiological dehydration and muscle wasting. Vipers exposed to dry air used more often a shelter that offered a moist microclimate, which reduced dehydration and muscle wasting at the individual level. These results provide the first experimental evidence that active behavioural hydroregulation can mitigate specific physiological stress responses caused by a dry spell in an ectotherm. Future studies investigating organismal responses to climate change should consider moisture gradient in the habitat and integrate both hydroregulation and thermoregulation behaviours.

Lizards from warm and declining populations are born with extremely short telomeres.

Aging is the price to pay for acquiring and processing energy through cellular activity and life history productivity. Climate warming can exacerbate the inherent pace of aging, as illustrated by a faster erosion of protective telomere DNA sequences. This biomarker integrates individual pace of life and parental effects through the germline, but whether intra- and intergenerational telomere dynamics underlies population trends remains an open question. Here, we investigated the covariation between life history, telomere length (TL), and extinction risk among three age classes in a cold-adapted ectotherm (Zootoca vivipara) facing warming-induced extirpations in its distribution limits. TL followed the same threshold relationships with population extinction risk at birth, maturity, and adulthood, suggesting intergenerational accumulation of accelerated aging rate in declining populations. In dwindling populations, most neonates inherited already short telomeres, suggesting they were born physiologically old and unlikely to reach recruitment. At adulthood, TL further explained females' reproductive performance, switching from an index of individual quality in stable populations to a biomarker of reproductive costs in those close to extirpation. We compiled these results to propose the aging loop hypothesis and conceptualize how climate-driven telomere shortening in ectotherms may accumulate across generations and generate tipping points before local extirpation.

Experimental Exposure to Tebuconazole Affects Metabolism and Body Condition in a Passerine Bird, the House Sparrow (Passer domesticus).

Triazole compounds are among the most widely used fungicides in agroecosystems to protect crops from potential fungal diseases. Triazoles are suspected to have an impact on nontarget species due to their interactions with nonfungal sterol synthesis, and wild birds are likely to be contaminated by triazole fungicides because many of them live in agroecosystems. We experimentally tested whether exposure to environmental concentrations of a triazole could alter key integrative traits (metabolic rates and body condition) of an agroecosystem sentinel species, the house sparrow (Passer domesticus). Wild-caught adult sparrows were maintained in captivity and exposed (exposed group) or not (control group) for 7 continuous months to tebuconazole through drinking water. The metabolic rates of exposed and control sparrows were then measured at two different temperatures (12 °C and 25 °C), which correspond, respectively, to the thermoregulation and thermoneutrality temperatures of this species. We found that exposed sparrows had lower resting metabolic rates (i.e., measured at thermoneutrality, 25 °C) than controls. However, the thermoregulatory metabolic rates (i.e., measured at 12 °C) did not differ between exposed and control sparrows. Although the body mass and condition were not measured at the beginning of the exposure, sparrows at the time of the metabolic measurements 7 months after the onset of such exposure had a higher body condition than controls, supporting further the idea that tebuconazole affects metabolic functions. Our study demonstrates for the first time that the use of tebuconazole can alter metabolism and could potentially lead to adverse effects in birds. Environ Toxicol Chem 2022;41:2500-2511. © 2022 SETAC.

Nature vs. Nurture: Disentangling the Influence of Inheritance, Incubation Temperature, and Post-Natal Care on Offspring Heart Rate and Metabolism in Zebra Finches.

A historic debate in biology is the question of nature vs. nurture. Although it is now known that most traits are a product of both heredity ("nature") and the environment ("nurture"), these two driving forces of trait development are rarely examined together. In birds, one important aspect of the early developmental environment is egg incubation temperature. Small changes (<1°C) in incubation temperature can have large effects on a wide-array of offspring traits. One important trait is metabolism, because it is related to life-history traits and strategies, organismal performance, and energetic and behavioral strategies. Although it has been shown that embryonic and post-hatch metabolism are related to egg incubation temperature, little is known about how this may vary as a function of genetic differences or post-hatching environmental conditions. Here, we investigated this question in zebra finches (Taeniopygia guttata). We experimentally incubated eggs at two different temperatures: 37.5°C (control), which is optimal for this species and 36.3°C (low), which is suboptimal. We first measured embryonic heart rate as a proxy of embryonic metabolic rate. Then, at hatch, we cross-fostered nestlings to differentiate genetic and pre-hatching factors from post-hatching environmental conditions. When offspring were 30 days-old, we measured their resting metabolic rate (RMR; within the thermoneutral zone) and thermoregulatory metabolic rate (TMR; 12°C; birds must actively thermoregulate). We also measured RMR and TMR of all genetic and foster parents. We found that embryonic heart rate was greater in eggs incubated at the control temperature than those at the low temperature. Further, embryonic heart rate was positively related to genetic father RMR, suggesting that it is both heritable and affected by the pre-natal environment. In addition, we found that post-hatch metabolic rates were positively related to genetic parent metabolic rate, and interactively related to incubation temperature and foster mother metabolic rate. Altogether, this suggests that metabolism and the energetic cost of thermoregulation can be influenced by genetics, the pre-natal environment, and the post-natal environment. Our study sheds light on how environmental changes and parental care may affect avian physiology, as well as which traits may be susceptible to natural selection.

Two stressors are worse than one: combined heatwave and drought affect hydration state and glucocorticoid levels in a temperate ectotherm.

Heatwaves and droughts are becoming more intense and frequent with climate change. These extreme weather events often occur simultaneously and may alter organismal physiology, yet their combined impacts remain largely unknown. Here, we experimentally investigated physiological responses of a temperate ectotherm, the asp viper (Vipera aspis), to a simulated heatwave and drought. We applied a two-by-two factorial design by manipulating the daily temperature cycle (control versus heatwave) and the water availability (water available versus water deprived) over a month followed by exposure to standard thermal conditions with ad libitum access to water. Simulated heatwave and water deprivation additively increased mass loss, while water deprivation led to greater plasma osmolality (dehydration). Mass gain from drinking after the treatment period was higher in vipers from the heatwave and water-deprived group, suggesting that thirst was synergistically influenced by thermal and water constraints. Heatwave conditions and water deprivation also additively increased baseline corticosterone levels but did not influence basal metabolic rate and plasma markers of oxidative stress. Our results demonstrate that a short-term exposure to combined heatwave and drought can exacerbate physiological stress through additive effects, and interactively impact behavioral responses to dehydration. Considering combined effects of temperature and water availability is thus crucial to assess organismal responses to climate change.

Agricultural intensification alters marbled newt genetic diversity and gene flow through density and dispersal reduction.

Recent agricultural intensification threatens global biodiversity with amphibians being one of the most impacted groups. Because of their biphasic life cycle, amphibians are particularly vulnerable to habitat loss and fragmentation that often result in small, isolated populations and loss of genetic diversity. Here, we studied how landscape heterogeneity affects genetic diversity, gene flow and demographic parameters in the marbled newt, Triturus marmoratus, over a hedgerow network landscape in Western France. While the northern part of the study area consists of preserved hedged farmland, the southern part was more profoundly converted for intensive arable crops production after WWII. Based on 67 sampled ponds and 10 microsatellite loci, we characterized regional population genetic structure and evaluated the correlation between landscape variables and (i) local genetic diversity using mixed models and (ii) genetic distance using multiple regression methods and commonality analysis. We identified a single genetic population characterized by a spatially heterogeneous isolation-by-distance pattern. Pond density in the surrounding landscape positively affected local genetic diversity while arable crop land cover negatively affected gene flow and connectivity. We used demographic inferences to quantitatively assess differences in effective population density and dispersal between the contrasted landscapes characterizing the northern and southern parts of the study area. Altogether, results suggest recent land conversion affected T. marmoratus through reduction in both effective population density and dispersal due to habitat loss and reduced connectivity.

Getting ready for a long bath: skin permeability decreases prior to aquatic breeding in male toads.

Vertebrate skin regulates exchanges between the organism and its environment and notably plays a fundamental role in regulating water fluxes. Dynamic changes of skin resistance to water fluxes are expected to occur in species that regularly shift between habitat types especially if these habitats differ in their hydric properties (e.g., terrestrial versus aquatic). We investigated changes of skin permeability using a study system (terrestrial toads) where reproduction induces a transition from terrestrial to freshwater habitats and a prolonged immersion that can last several weeks in males. In this system, the simultaneity between skin shedding and the onset of breeding suggests that the production of new integument layers prior to immersion for reproduction may regulate water influxes. We found that the skin permeability of male toads decreases significantly prior to breeding, suggesting that skin shedding at the onset of breeding regulates water fluxes to alleviate osmotic costs of immersion during reproduction. The continued decrease of skin permeability detected during breeding suggests that additional mechanisms interact with skin structure to further decrease permeability to water during a prolonged immersion. Future studies are required to assess whether changes in skin permeability to water tradeoffs with other skin characteristics (gas exchanges) relevant to aquatic breeding amphibians.

Water deprivation compromises maternal physiology and reproductive success in a cold and wet adapted snake Vipera berus.

Droughts are becoming more intense and frequent with climate change. These extreme weather events can lead to mass mortality and reproduction failure, and therefore cause population declines. Understanding how the reproductive physiology of organisms is affected by water shortages will help clarify whether females can adjust their reproductive strategy to dry conditions or may fail to reproduce and survive. In this study, we investigated the consequences of a short period of water deprivation (2 weeks) during early pregnancy on the physiology and behaviour of a cold- and wet-adapted ectotherm (Vipera berus). We also examined water allocation to developing embryos and embryonic survival. Water-deprived females exhibited significant dehydration, physiological stress and loss of muscle mass. These effects of water deprivation on water balance and muscle loss were correlated with the number of developing embryos. While water-deprived females maintained water transfer to embryos at the expense of their own maintenance, water deprivation also led to embryonic mortality. Overall, water deprivation amplifies the reproductive costs of water allocation to support embryonic development. The deleterious impacts of water deprivation on female current reproductive performance and on potential survival and future reproduction could lead to severe population declines in this species.

Determinants of Legacy Persistent Organic Pollutant Levels in the European Pond Turtle (Emys orbicularis) in the Camargue Wetland, France.

Many banned persistent organic pollutants (POPs) remain for decades in the aquatic environment and can have harmful effects on long-lived predators because of their high bioaccumulation and biomagnification potentials. We investigated the occurrence and levels of 18 polychlorinated biphenyls (PCBs) and 16 organochlorine pesticides in European pond turtles (n = 174) from April to July 2018 in the Camargue wetland, France. Although the Camargue was highly contaminated in previous decades, plasma occurrence and levels of POPs were very low: we were able to quantify only 3 of the 34 compounds we analyzed in >10% of the turtles. The burdens from POPs did not differ between males and females and were uncorrelated with sampling date and body mass. We observed differences in POP burdens between turtles from the 2 sampling sites. One possible explanation is that the sampling sites were in different agricultural hydraulic systems: plasma occurrence and levels were higher for PCB-52 and hexachlorobenzene in turtles captured in drainage channels and for PCB-153 at the site that receives irrigation. Finally, the occurrence and levels of PCB-153 in turtles increased with age, likely because of bioaccumulation and much higher exposure 20 to 30 yr ago than now. Environ Toxicol Chem 2021;40:2261-2268. © 2021 SETAC.

Additive effects of temperature and water availability on pregnancy in a viviparous lizard.

One of the greatest current threats to biodiversity is climate change. However, understanding of organismal responses to fluctuations in temperature and water availability is currently lacking, especially during fundamental life-history stages such as reproduction. To further explore how temperature and water availability impact maternal physiology and reproductive output, we used the viviparous form of the European common lizard (Zootoca vivipara) in a two-by-two factorial design manipulating both hydric and thermal conditions, for the first time. We collected blood samples and morphological measurements during early pregnancy and post-parturition to investigate how water availability, temperature and a combination of the two influence maternal phenology, morphology, physiology and reproductive output. We observed that dehydration during gestation negatively affects maternal physiological condition (lower mass gain, higher tail reserve mobilization) but has little effect on reproductive output. These effects are mainly additive to temperature regimes, with a proportional increase in maternal costs in warmer environments. Our study demonstrates the importance of considering combined effects of water and temperature when investigating organismal responses to climate changes, especially during periods crucial for species survival such as reproduction.

Reproductive state and water deprivation increase plasma corticosterone in a capital breeder.

Plasma corticosterone (CORT) concentrations fluctuate in response to homeostatic demands. CORT is widely recognized as an important hormone related to energy balance. However, far less attention has been given to the potential role of CORT in regulating salt and water balance or responding to osmotic imbalances. We examined the effects of reproductive and hydric states on CORT levels in breeding Children's pythons (Antaresia childreni), a species with substantial energetic and hydric costs associated with egg development. Using a 2 × 2 experimental design, we examined how reproduction and water deprivation, both separately and combined, impact CORT levels and how these changes correlate with hydration (plasma osmolality) and energy levels (blood glucose). We found that reproduction leads to increased CORT levels, as does dehydration induced by water deprivation. The combined impact of reproduction and water deprivation led to the largest increases in CORT levels. Additionally, we found significant positive relationships among CORT levels, plasma osmolality, and blood glucose. Our results provide evidence that both reproductive activity and increased plasma osmolality can lead to increased plasma CORT in an ectotherm, which could be explained by either CORT having a role as a mineralocorticoid or CORT being elevated as part of a stress response to resource imbalances.

Slowing down the metabolic engine: impact of early-life corticosterone exposure on adult metabolism in house sparrows (Passer domesticus).

Whole-organism metabolism is an integrative process that determines not only the energy cost of living but also the energy output that is available for behavioral and physiological processes during the life cycle. Developmental challenge is known to affect growth, development of several organs, and several physiological mechanisms (such as HPA responsiveness, oxidative stress or immunity), which may altogether affect adult metabolism. All of these developmental effects are likely to be mediated by glucocorticoids, but the impact of developmental glucocorticoid exposure on adult metabolism has rarely been studied and the results are equivocal. In this study, we examined the impact of developmental exposure to corticosterone (CORT, the main avian glucocorticoid hormone) on resting metabolic rate (RMR, measured in thermoneutrality, 25°C) and thermoregulatory metabolic rate (TMR, measured in cold challenge conditions, 5°C) in the house sparrow. Following experimental administration of CORT at the nestling stage, house sparrows were kept in captivity until adulthood, when their metabolism was measured. We found that post-natal CORT exposure decreased both RMR and TMR in adult sparrows. This CORT-mediated reduction of metabolism was also associated with a reduced overnight body mass loss. Therefore, our results suggest that developmental CORT exposure can orient the phenotype towards an energy-saving strategy, which may be beneficial in a constraining environmental context.

When water interacts with temperature: Ecological and evolutionary implications of thermo-hydroregulation in terrestrial ectotherms.

The regulation of body temperature (thermoregulation) and of water balance (defined here as hydroregulation) are key processes underlying ecological and evolutionary responses to climate fluctuations in wild animal populations. In terrestrial (or semiterrestrial) ectotherms, thermoregulation and hydroregulation closely interact and combined temperature and water constraints should directly influence individual performances. Although comparative physiologists traditionally investigate jointly water and temperature regulation, the ecological and evolutionary implications of these coupled processes have so far mostly been studied independently. Here, we revisit the concept of thermo-hydroregulation to address the functional integration of body temperature and water balance regulation in terrestrial ectotherms. We demonstrate how thermo-hydroregulation provides a framework to investigate functional adaptations to joint environmental variation in temperature and water availability, and potential physiological and/or behavioral conflicts between thermoregulation and hydroregulation. We extend the classical cost-benefit model of thermoregulation in ectotherms to highlight the adaptive evolution of optimal thermo-hydroregulation strategies. Critical gaps in the parameterization of this conceptual optimality model and guidelines for future empirical research are discussed. We show that studies of thermo-hydroregulation refine our mechanistic understanding of physiological and behavioral plasticity, and of the fundamental niche of the species. This is illustrated with relevant and recent examples of space use and dispersal, resource-based trade-offs, and life-history tactics in insects, amphibians, and nonavian reptiles.

Some like it dry: Water restriction overrides heterogametic sex determination in two reptiles.

ABSTRACT: The evolution of sex determination is complex and yet crucial in our understanding of population stability. In ectotherms, sex determination involves a variety of mechanisms including genetic determination (GSD), environment determination (ESD), but also interactions between the two via sex reversal. In this study, we investigated whether water deprivation during pregnancy could override GSD in two heterogametic squamate reptiles. We demonstrated that water restriction in early gestation induced a male-biased secondary sex ratio in both species, which could be explained by water sex reversal as the more likely mechanism. We further monitored some long-term fitness estimates of offspring, which suggested that water sex determination (WSD) represented a compensatory strategy producing the rarest sex according to Fisher's assumptions of frequency-dependent selection models. This study provides new insights into sex determination modes and calls for a general investigation of mechanisms behind WSD and to examine the evolutionary implications. OPEN RESEARCH BADGES: This article has earned an Open Data Badge for making publicly available the digitally-shareable data necessary to reproduce the reported results. The data is available at https://doi.org/10.5061/dryad.mv06pv1.

Author Correction: Agricultural landscapes and the Loire River influence the genetic structure of the marbled newt in Western France.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has been fixed in the paper.