Reading tea leaves worldwide: Decoupled drivers of initial litter decomposition mass-loss rate and stabilization.

The breakdown of plant material fuels soil functioning and biodiversity. Currently, process understanding of global decomposition patterns and the drivers of such patterns are hampered by the lack of coherent large-scale datasets. We buried 36,000 individual litterbags (tea bags) worldwide and found an overall negative correlation between initial mass-loss rates and stabilization factors of plant-derived carbon, using the Tea Bag Index (TBI). The stabilization factor quantifies the degree to which easy-to-degrade components accumulate during early-stage decomposition (e.g. by environmental limitations). However, agriculture and an interaction between moisture and temperature led to a decoupling between initial mass-loss rates and stabilization, notably in colder locations. Using TBI improved mass-loss estimates of natural litter compared to models that ignored stabilization. Ignoring the transformation of dead plant material to more recalcitrant substances during early-stage decomposition, and the environmental control of this transformation, could overestimate carbon losses during early decomposition in carbon cycle models.

Aspectos da ecologia reprodutiva de Trachycephalus cunauaru (Anura: Hylidae) na Amazônia meridional / Aspects of the reproductive ecology of Trachycephalus cunauaru (Anura: Hylidae) in the southern Amazon

Trachycephalus cunauaru é um hilídeo amazônico que utiliza fitotelmatas para se reproduzir. Existem relativamente poucas informações sobre a espécie, principalmente devido à dificuldade de acesso aos seus sítios reprodutivos. Nesse trabalho, reunimos dados de ecologia e história natural de T. cunauaru em uma área da Amazônia meridional, no estado de Mato Grosso, Brasil. Além de fitotelmatas naturais, utilizamos baldes instalados a uma altura de 10 m como fitotelmatas artificiais. Comparamos características físicas e químicas, bem como a presença e ausência de girinos, entre fitotelmatas naturais e artificiais. Também coletamos dados sobre o comportamento reprodutivo da espécie por meio de armadilhas fotográficas. Registramos uma densidade de 14,1 sítios reprodutivos por km2 . Os parâmetros ambientais diferiram significativamente entre fitotelmatas artificiais e naturais. Em sítios artificiais a presença de girinos esteve diretamente relacionada à árvores com maior diâmetro. Registramos, pela primeira vez, oofagia para a espécie e observamos que machos podem utilizar mais de um fitotelmata. Também registramos a presença de cobras nos sítios reprodutivos. Constatamos que sítios artificiais e armadilhas fotográficas representam uma alternativa satisfatória para registros comportamentais para T. cunauaru e, possivelmente, para outras espécies com hábitos similares. (AU)

Rate-Pressure Product Responses to Static Contractions Performed at Various Altitudes.

Simmonds, Michael J., Surendran Sabapathy, and Jean-Marc Hero. Rate-pressure product responses to static contractions performed at various altitudes. High Alt Med Biol. 22: 166-173, 2021. Background: Adventure tourism has led to an unprecedented number of individuals being exposed to altitude, including those with subclinical cardiometabolic disorders. The disproportionate hemodynamic challenge associated with small-muscle static activities is potentially dangerous at altitude as these may compound the risk for cardiac events. We thus examined the cardiovascular response to, and during recovery from, static exercise performed at altitude. Methods: Eighteen individuals completed this study at three altitudes (sea level; ∼1,500 m; ∼3,000 m) in central Nepal. At each altitude, individuals performed two handgrip contractions for 2 minutes at the same intensity (30% maximal voluntary contraction [MVC]), with two distinct recovery periods: during control recovery was completed quietly at rest, while during ischemic challenge recovery was conducted with a cuff occluding the upper limb. Results: Oxygen saturation decreased during ascent to 1,500 m (-2%) and 3,000 m (-8%), compared with sea level. Handgrip MVC was not affected by altitude, although heart rate at rest (∼70 beat/min), during static exercise (range ∼90-95 beat/min), and during recovery in both conditions (each ∼70 beat/min) was significantly increased by ∼15% at 3,000 m, but not 1,500 m. The magnitude of the muscle metaboreflex during recovery from static exercise was unaffected by altitude; however, the rate-pressure product was significantly elevated by ∼10% during and following static exercise at 3,000 m. Conclusions: A significant increase in the rate-pressure product during static exercise was observed at altitude, which persisted during recovery. Individuals at risk for cardiac events should use awareness of static contractions while at altitude, especially considering that stress induced by static exercise is additive to that of dynamic activities such as hiking.

Research priorities for natural ecosystems in a changing global climate.

Climate change poses significant emerging risks to biodiversity, ecosystem function and associated socioecological systems. Adaptation responses must be initiated in parallel with mitigation efforts, but resources are limited. As climate risks are not distributed equally across taxa, ecosystems and processes, strategic prioritization of research that addresses stakeholder-relevant knowledge gaps will accelerate effective uptake into adaptation policy and management action. After a decade of climate change adaptation research within the Australian National Climate Change Adaptation Research Facility, we synthesize the National Adaptation Research Plans for marine, terrestrial and freshwater ecosystems. We identify the key, globally relevant priorities for ongoing research relevant to informing adaptation policy and environmental management aimed at maximizing the resilience of natural ecosystems to climate change. Informed by both global literature and an extensive stakeholder consultation across all ecosystems, sectors and regions in Australia, involving thousands of participants, we suggest 18 priority research topics based on their significance, urgency, technical and economic feasibility, existing knowledge gaps and potential for cobenefits across multiple sectors. These research priorities provide a unified guide for policymakers, funding organizations and researchers to strategically direct resources, maximize stakeholder uptake of resulting knowledge and minimize the impacts of climate change on natural ecosystems. Given the pace of climate change, it is imperative that we inform and accelerate adaptation progress in all regions around the world.

Gut microbiota and their putative metabolic functions in fragmented Bengal tiger population of Nepal.

Bengal tigers (Panthera tigris tigris) serve a pivotal role as an apex predator in forest ecosystems. To increase our knowledge on factors impacting the viability and health of this endangered species, we studied the gut microbiota in 32 individual Bengal tigers from three geographically separated areas (Chitwan National Park (CNP), Bardia National Park (BNP) and Suklaphanta Wildlife Reserve (SWR)) in Nepal, using noninvasive genetic sampling methods. Gut microbiota influence the immune system, impact various physiological functions, and modulates metabolic reactions, that ultimately impact the host health, behavior and development. Across the tiger populations in Nepal, we found significant differences in the composition of microbial communities based on their geographic locations. Specifically, we detected significant differences between CNP and the other two protected areas (CNP vs BNP: pseudo t = 1.944, P = 0.006; CNP vs SWR: pseudo t = 1.9942, P = 0.0071), but no differences between BNP and SWR. This mirrors what has been found for tiger gene flow in the same populations, suggesting gut microbiota composition and host gene flow may be linked. Furthermore, predictive metagenome functional content analysis (PICRUSt) revealed a higher functional enrichment and diversity for significant gut microbiota in the Chitwan tiger population and the lowest enrichment and diversity in Suklaphanta. The CNP tiger population contained higher proportions of microbiota that are associated with predicted functions relevant for metabolism of amino acid, lipid, xenobiotics biodegradation, terpenoides and polyketides than the SWR population. We conclude the tiger population structure, gut microbiota profile and associated functional metabolic categories are correlated, with geographically most separated CNP and SWR tiger population having the most distinct and different host genotype and microbiota profiles. Our work dramatically expands the understanding of tiger microbiota in wild populations and provides a valuable case study on how to investigate genetic diversity at different hierarchical levels, including hosts as well as their microbial communities.

Correction: Surveillance of Influenza A Virus and Its Subtypes in Migratory Wild Birds of Nepal.

[This corrects the article DOI: 10.1371/journal.pone.0133035.].

Species, sex and geo-location identification of seized tiger (Panthera tigris tigris) parts in Nepal-A molecular forensic approach.

Tiger (Panthera tigris) populations are in danger across their entire range due to habitat loss, poaching and the demand for tiger parts. The Bengal tiger (Panthera tigris tigris) is an endangered apex predator with a population size estimated to be less than 200 in Nepal. In spite of strict wildlife protection laws, illegal trade of tiger parts is increasing; and Nepal has become one of the major sources and transit routes for poached wildlife parts. Identification of wildlife parts is often challenging for law enforcement officials due to inadequate training and lack of available tools. Here, we describe a molecular forensic approach to gain insight into illegally trafficked tiger parts seized across Nepal. We created Nepal's first comprehensive reference genetic database of wild tigers through the Nepal Tiger Genome Project (2011-2013). This database has nuclear DNA microsatellite genotype and sex profiles, including geo-spatial information, of over 60% (n = 120) of the wild tigers of Nepal. We analyzed 15 putative cases of confiscated poached tiger parts and all were confirmed to be of tiger. Ten samples were identified as male and five were female. We determined probable geo-source location for 9 of the 14 samples with 6-8 nuclear DNA microsatellite loci using inferences from four different statistical assignment methods. Six samples were assigned to Bardia National Park and one of these was an exact match to a female tiger previously profiled in our fecal DNA reference database. Two tiger samples were assigned to Shuklaphanta Wildlife Reserve and one to Chitwan National Park. We are unable to definitively assign five tiger samples which could be offspring dispersers or might have come from tiger population outside of Nepal. Our study revealed that the western region, particularly Bardia National Park, is a poaching hotspot for illegal tiger trade in Nepal. We present feasibility of using molecular forensic based evidence to incriminate criminals in a court of law in the fight against wildlife crime.

Assessment of genetic diversity, population structure, and gene flow of tigers (Panthera tigris tigris) across Nepal's Terai Arc Landscape.

With fewer than 200 tigers (Panthera tigris tigris) left in Nepal, that are generally confined to five protected areas across the Terai Arc Landscape, genetic studies are needed to provide crucial information on diversity and connectivity for devising an effective country-wide tiger conservation strategy. As part of the Nepal Tiger Genome Project, we studied landscape change, genetic variation, population structure, and gene flow of tigers across the Terai Arc Landscape by conducting Nepal's first comprehensive and systematic scat-based, non-invasive genetic survey. Of the 770 scat samples collected opportunistically from five protected areas and six presumed corridors, 412 were tiger (57%). Out of ten microsatellite loci, we retain eight markers that were used in identifying 78 individual tigers. We used this dataset to examine population structure, genetic variation, contemporary gene flow, and potential population bottlenecks of tigers in Nepal. We detected three genetic clusters consistent with three demographic sub-populations and found moderate levels of genetic variation (He = 0.61, AR = 3.51) and genetic differentiation (FST = 0.14) across the landscape. We detected 3-7 migrants, confirming the potential for dispersal-mediated gene flow across the landscape. We found evidence of a bottleneck signature likely caused by large-scale land-use change documented in the last two centuries in the Terai forest. Securing tiger habitat including functional forest corridors is essential to enhance gene flow across the landscape and ensure long-term tiger survival. This requires cooperation among multiple stakeholders and careful conservation planning to prevent detrimental effects of anthropogenic activities on tigers.

Proximate causes of altitudinal differences in body size in an agamid lizard.

Body size is directly linked to key life history traits such as growth, fecundity, and survivorship. Identifying the causes of body size variation is a critical task in ecological and evolutionary research. Body size variation along altitudinal gradients has received considerable attention; however, the underlying mechanisms are poorly understood. Here, we compared the growth rate and age structure of toad-headed lizards (Phrynocephalus vlangalii) from two populations found at different elevations in the Qinghai-Tibetan Plateau. We used mark-recapture and skeletochronological analysis to identify the potential proximate causes of altitudinal variation in body size. Lizards from the high-elevation site had higher growth rates and attained slightly larger adult body sizes than lizards from the low-elevation site. However, newborns produced by high-elevation females were smaller than those by low-elevation females. Von Bertalanffy growth estimates predicted high-elevation individuals would reach sexual maturity at an earlier age and have a lower mean age than low-elevation individuals. Relatively lower mean age for the high-elevation population was confirmed using the skeletochronological analysis. These results support the prediction that a larger adult body size of high-elevation P. vlangalii results from higher growth rates, associated with higher resource availability.

Calling phenology of a diverse amphibian assemblage in response to meteorological conditions.

The strong association between amphibian activity, breeding and recruitment with local environmental conditions raises concerns regarding how changes in climate may affect the persistence of species populations into the future. Additionally, in a highly diverse assemblage of anurans, competition for breeding sites affects the time and duration of activity, as species compete for limited resources such as water. Meteorological conditions are strong drivers of amphibian activity, so we assessed whether temperature, rainfall, atmospheric pressure and humidity were associated with the calling phenology of an assemblage of anurans in South East Queensland, Australia. We performed calling surveys and collected digital recordings at 45 ponds in an area known for high anuran diversity. We performed detection analyses to investigate the influence of 10 meteorological variables in detection of calling activity in 19 amphibian species. Our results suggest four breeding strategies in the assemblage: explosive summer breeders, prolonged breeders, opportunistic breeders and a winter breeder. Classifying these species into associations provides a framework for understanding how species respond to environmental conditions. Explosive breeders (i.e. species demonstrating short and highly synchronised breeding periods) were particularly responsive to temperature. Our findings help elucidate the breeding phenology of frogs and provide valuable information on their mating systems in native Australian forests. This study highlights the difficulties of surveying even common anurans. We highlight the importance of predictability and stability in climate and the vulnerability of species for which reproduction appears to require highly specific environmental cues.

Visualizing Phonotactic Behavior of Female Frogs in Darkness.

Many animals use sounds produced by conspecifics for mate identification. Female insects and anuran amphibians, for instance, use acoustic cues to localize, orient toward and approach conspecific males prior to mating. Here we present a novel technique that utilizes multiple, distributed sound-indication devices and a miniature LED backpack to visualize and record the nocturnal phonotactic approach of females of the Australian orange-eyed tree frog (Litoria chloris) both in a laboratory arena and in the animal's natural habitat. Continuous high-definition digital recording of the LED coordinates provides automatic tracking of the female's position, and the illumination patterns of the sound-indication devices allow us to discriminate multiple sound sources including loudspeakers broadcasting calls as well as calls emitted by individual male frogs. This innovative methodology is widely applicable for the study of phonotaxis and spatial structures of acoustically communicating nocturnal animals.

Does physiological response to disease incur cost to reproductive ecology in a sexually dichromatic amphibian species?

It is well known that the disease chytridiomycosis, caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd) has contributed to amphibian declines worldwide. The impact of Bd varies, with some species being more susceptible to infection than others. Recent evidence has shown that Bd can have sub-lethal effects, whereby increases in stress hormones have been associated with infection. Could this increased stress response, which is a physiological adaptation that provides an increased resilience against Bd infection, potentially be a trade-off with important life-history traits such as reproduction? We studied this question in adult male frogs of a non-declining species (Litoria wilcoxii). Frogs were sampled for (1) seasonal hormone (testosterone and corticosterone), color and disease profiles, (2) the relationship between disease infection status and hormone levels or dorsal color, (3) subclinical effects of Bd by investigating disease load and hormone level, and (4) reproductive and stress hormone relationships independent of disease. Testosterone levels and color score varied seasonally (throughout the spring/summer months) while corticosterone levels remained stable. Frogs with high Bd prevalence had significantly higher corticosterone levels and lower testosterone levels compared to uninfected frogs, and no differences in color were observed. There was a significant positive correlation between disease load and corticosterone levels, and a significant negative relationship between disease load and testosterone. Our field data provides novel evidence that increased physiological stress response associated with Bd infection in wild frogs, could suppress reproduction by down-regulating gonadal hormones in amphibians, however the impacts on reproductive output is yet to be established.

Temporal Variation of the Skin Bacterial Community and Batrachochytrium dendrobatidis Infection in the Terrestrial Cryptic Frog Philoria loveridgei.

In animals and plants, symbiotic bacteria can play an important role in disease resistance of host and are the focus of much current research. Globally, amphibian population declines and extinctions have occurred due to chytridiomycosis, a skin disease caused by the pathogen Batrachochytrium dendrobatidis (Bd). Currently amphibian skin bacteria are increasingly recognized as important symbiont communities with a relevant role in the defense against pathogens, as some bacteria can inhibit the growth of B. dendrobatidis. This study aims to document the B. dendrobatidis infection status of wild populations of a terrestrial cryptic frog (Philoria loveridgei), and to determine whether infection status is correlated with changes in the skin microbial communities. Skin samples of P. loveridgei were collected along an altitudinal range within the species distribution in subtropical rainforests in southeast Australia. Sampling was conducted in two years during two breeding seasons with the first classified as a "La Niña" year. We used Taqman real-time PCR to determine B. dendrobatidis infection status and 16S amplicon sequencing techniques to describe the skin community structure. We found B. dendrobatidis-positive frogs only in the second sampling year with low infection intensities, and no correlation between B. dendrobatidis infection status and altitude, frog sex or size. Skin bacterial diversity was significantly higher in P. loveridgei frogs sampled in the 1st year than in the 2nd year. In addition, 7.4% of the total OTUs were significantly more abundant in the 1st year compared to the 2nd year. We identified 67 bacterial OTUs with a significant positive correlation between infection intensity and an OTU's relative abundance. Forty-five percent of these OTUs belonged to the family Enterobacteriaceae. Overall, temporal variation was strongly associated with changes in B. dendrobatidis infection status and bacterial community structure of wild populations of P. loveridgei.

The Australian SuperSite Network: A continental, long-term terrestrial ecosystem observatory.

Ecosystem monitoring networks aim to collect data on physical, chemical and biological systems and their interactions that shape the biosphere. Here we introduce the Australian SuperSite Network that, along with complementary facilities of Australia's Terrestrial Ecosystem Research Network (TERN), delivers field infrastructure and diverse, ecosystem-related datasets for use by researchers, educators and policy makers. The SuperSite Network uses infrastructure replicated across research sites in different biomes, to allow comparisons across ecosystems and improve scalability of findings to regional, continental and global scales. This conforms with the approaches of other ecosystem monitoring networks such as Critical Zone Observatories, the U.S. National Ecological Observatory Network; Analysis and Experimentation on Ecosystems, Europe; Chinese Ecosystem Research Network; International Long Term Ecological Research network and the United States Long Term Ecological Research Network. The Australian SuperSite Network currently involves 10 SuperSites across a diverse range of biomes, including tropical rainforest, grassland and savanna; wet and dry sclerophyll forest and woodland; and semi-arid grassland, woodland and savanna. The focus of the SuperSite Network is on using vegetation, faunal and biophysical monitoring to develop a process-based understanding of ecosystem function and change in Australian biomes; and to link this with data streams provided by the series of flux towers across the network. The Australian SuperSite Network is also intended to support a range of auxiliary researchers who contribute to the growing body of knowledge within and across the SuperSite Network, public outreach and education to promote environmental awareness and the role of ecosystem monitoring in the management of Australian environments.

Surveillance of Influenza A Virus and Its Subtypes in Migratory Wild Birds of Nepal.

Nepal boarders India and China and all three countries lie within the Central Asian Flyway for migratory birds. Novel influenza A H7N9 caused human fatalities in China in 2013. Subclinical infections of influenza A H7N9 in birds and the potential for virus dispersal by migratory birds prompted this study to assess avian H7N9 viral intrusion into Nepal. Surveillance of influenza A virus in migratory birds was implemented in early 2014 with assistance from the Food and Agricultural Organization (FAO). Of 1811 environmental fecal samples collected from seven wetland migratory bird roosting areas, influenza A H9N2 was found in one sample from a ruddy shelduck in Koshi Tappu Wildlife Reserve located in southern Nepal. Avian H7N9 and other highly pathogenic avian influenza viruses were not detected. This study provides baseline data on the status of avian influenza virus in migratory bird populations in Nepal.

Maximizing the reliability of non-invasive endocrine sampling in the tiger (Panthera tigris): environmental decay and intra-sample variation in faecal glucocorticoid metabolites.

Evaluation of physiological stress in the tiger (Panthera tigris) using faecal cortisol metabolite (FCM) enzyme immunoassays (EIAs) provides a powerful conservation physiology tool for the species. However, it is important to validate non-invasive endocrine sampling techniques in field conditions to ensure that the method provides a reliable parameter of physiological stress in the species. This is because endocrine measurements are highly species specific and FCM concentrations can be influenced by environmental factors. Here, we studied the impact of the decay rate of FCMs and intra-sample variation of FCMs using a previously validated EIA. To determine the decay rate of FCMs, we measured FCMs in freshly deposited tiger faeces (n = 8 tigers and 48 scats) that were randomly exposed to the natural environment (dry conditions with no rainfall) for up to 192 h. To determine intra-sample variation in FCMs, we used 10 scats from 10 tigers, divided each sample into four sections and each section into four sub-sections and measured FCMs in each section and sub-section. The results of this decay-rate experiment showed that FCMs in tiger faeces began to decay after 48 h exposure to the environmental conditions available. Thus, FCMs within freshly deposited tiger faeces are influenced by available environmental conditions. Changes in weather conditions (e.g. increased rainfall and humidity) could influence the stability of FCMs. The results of the intra-sample variation study showed that inter-variation among scats accounted for 52% of the variations in FCMs, while intra-sample variation between sections (32%) was greater than the sub-sample variation (16%). Intra-sample variation can be reduced by homogenizing the entire lyophilized faecal sample prior to the EIA. In conclusion, careful evaluation of decay rate and complete homogenization of faeces prior to EIA analysis will increase the reliability of FCMs as a non-invasive index of physiological stress in the tiger.

The neuro-hormonal control of rapid dynamic skin colour change in an amphibian during amplexus.

Sexual signalling using dynamic skin colouration is a key feature in some vertebrates; however, it is rarely studied in amphibians. Consequently, little is known about the hormonal basis of this interesting biological phenomenon for many species. Male stony creek frogs (Litoria wilcoxii) are known to change dorsal colouration from brown to lemon yellow within minutes. This striking change is faster then what has been seen most amphibians, and could therefore be under neuronal regulation, a factor that is rarely observed in amphibians. In this study, we observed colour changes in wild frogs during amplexus to determine the natural timing of colour change. We also investigated the hypothesis that colour change is mediated by either reproductive or neuro- hormones. This was achieved by injecting frogs with epinephrine, testosterone, saline solution (control 1) or sesame oil (control 2). A non-invasive approach was also used wherein hormones and controls were administered topically. Male frogs turned a vivid yellow within 5 minutes of initiation of amplexus and remained so for 3-5 hours before rapidly fading back to brown. Epinephrine-treated frogs showed a significant colour change from brown to yellow within 5 minutes, however, testosterone-treated frogs did not change colour. Our results provide evidence of the role neuronal regulation plays in colour change systems.

Determinants of tree assemblage composition at the mesoscale within a subtropical eucalypt forest.

A variety of environmental processes, including topography, edaphic and disturbance factors can influence vegetation composition. The relative influence of these patterns has been known to vary with scale, however, few studies have focused on environmental drivers of composition at the mesoscale. This study examined the relative importance of topography, catchment flow and soil in influencing tree assemblages in Karawatha Forest Park; a South-East Queensland subtropical eucalypt forest embedded in an urban matrix that is part of the Terrestrial Ecosystem Research Network South-East Queensland Peri-urban SuperSite. Thirty-three LTER plots were surveyed at the mesoscale (909 ha), where all woody stems ≥1.3 m high rooted within plots were sampled. Vegetation was divided into three cohorts: small (≥1-10 cm DBH), intermediate (≥10-30 cm DBH), and large (≥30 cm DBH). Plot slope, aspect, elevation, catchment area and location and soil chemistry and structure were also measured. Ordinations and smooth surface modelling were used to determine drivers of vegetation assemblage in each cohort. Vegetation composition was highly variable among plots at the mesoscale (plots systematically placed at 500 m intervals). Elevation was strongly related to woody vegetation composition across all cohorts (R2: 0.69-0.75). Other topographic variables that explained a substantial amount of variation in composition were catchment area (R2: 0.43-0.45) and slope (R2: 0.23-0.61). Soil chemistry (R2: 0.09-0.75) was also associated with woody vegetation composition. While species composition differed substantially between cohorts, the environmental variables explaining composition did not. These results demonstrate the overriding importance of elevation and other topographic features in discriminating tree assemblage patterns irrespective of tree size. The importance of soil characteristics to tree assemblages was also influenced by topography, where ridge top sites were typically drier and had lower soil nutrient levels than riparian areas.

Repeated thermal stressor causes chronic elevation of baseline corticosterone and suppresses the physiological endocrine sensitivity to acute stressor in the cane toad (Rhinella marina).

Extreme environmental temperature could impact the physiology and ecology of animals. The stress endocrine axis provides necessary physiological stress response to acute (day-day) stressors. Presently, there are no empirical evidences showing that exposure to extreme thermal stressor could cause chronic stress in amphibians. This could also modulate the physiological endocrine sensitivity to acute stressors and have serious implications for stress coping in amphibians, particularly those living in fragmented and disease prone environments. We addressed this important question using the cane toad (Rhinella marina) model from its introduced range in Queensland, Australia. We quantified their physiological endocrine sensitivity to a standard acute (capture and handling) stressor after exposing the cane toads to thermal shock at 35°C for 30min daily for 34 days. Corticosterone (CORT) responses to the capture and handling protocol were measured on three sampling intervals (days 14, 24, and 34) to determine whether the physiological endocrine sensitivity was maintained or modulated over-time. Two control groups (C1 for baseline CORT measurement only and C2 acute handled only) and two temperature treatment groups (T1 received daily thermal shock up to day 14 only and a recovery phase of 20 days and T2 received thermal shock daily for 34 days). Results showed that baseline CORT levels remained high on day 14 (combined effect of capture, captivity and thermal stress) for both T1 and T2. Furthermore, baseline CORT levels decreased for T1 once the thermal shock was removed after day 14 and returned to baseline by day 29. On the contrary, baseline CORT levels kept on increasing for T2 over the 34 days of daily thermal shocks. Furthermore, the magnitudes of the acute CORT responses or physiological endocrine sensitivity were consistently high for both C1 and T1. However, acute CORT responses for T2 toads were dramatically reduced between days 24 and 34. These novel findings suggest that repeated exposure to extreme thermal stressor could cause chronic stress and consequently suppress the physiological endocrine sensitivity to acute stressors (e.g. pathogenic diseases) in amphibians.

Acute thermal stressor increases glucocorticoid response but minimizes testosterone and locomotor performance in the cane toad (Rhinella marina).

Climatic warming is a global problem and acute thermal stressor in particular could be considered as a major stressor for wildlife. Cane toads (Rhinella marina) have expanded their range into warmer regions of Australia and they provide a suitable model species to study the sub-lethal impacts of thermal stressor on the endocrine physiology of amphibians. Presently, there is no information to show that exposure to an acute thermal stressor could initiate a physiological stress (glucocorticoid) response and secondly, the possible effects on reproductive hormones and performance. Answering these questions is important for understanding the impacts of extreme temperature on amphibians. In this study, we experimented on cane toads from Queensland, Australia by acclimating them to mildly warm temperature (25°C) and then exposing to acute temperature treatments of 30°, 35° or 40°C (hypothetical acute thermal stressors). We measured acute changes in the stress hormone corticosterone and the reproductive hormone testosterone using standard capture and handling protocol and quantified the metabolites of both hormones non-invasively using urinary enzyme-immunoassays. Furthermore, we measured performance trait (i.e. righting response score) in the control acclimated and the three treatment groups. Corticosterone stress responses increased in all toads during exposure to an acute thermal stressor. Furthermore, exposure to a thermal stressor also decreased testosterone levels in all toads. The duration of the righting response (seconds) was longer for toads that were exposed to 40°C than to 30°, 35° or 25°C. The increased corticosterone stress response with increased intensity of the acute thermal stressor suggests that the toads perceived this treatment as a stressor. Furthermore, the results also highlight a potential trade-off with performance and reproductive hormones. Ultimately, exposure acute thermal stressors due to climatic variability could impact amphibians at multiple eco-physiological levels through impacts on endocrine physiology, performance and potentially fitness traits (e.g. reproductive output).