The impact of health and disease on wildlife population dynamics and individual survival is complex and poorly understood, especially in cryptic species such as chelonians. Eastern box turtles (Terrapene carolina carolina) are declining due to anthropogenic and natural factors, including disease, though the relative importance of these factors for individual survival is unknown. Determining survival rates in free-ranging chelonians is challenging because individuals are difficult to locate and recapture, deceased turtles can be quickly scavenged, and turtles can die underground during a brumation period. The purpose of this study was to estimate the apparent survival rate for wild eastern box turtles detected with common box turtle pathogens, including Terrapene herpesvirus 1, Terrapene adenovirus, and box turtle Mycoplasmopsis sp., using Cormack-Jolly-Seber models. We used mark-recapture data from 778 individuals from five box turtle populations collected over 7 yr (2016-22), paired with concurrently collected demographic and quantitative PCR pathogen detection data. Apparent survival estimates were different among the five sites, ranging from 71% to 88%, but similar between sexes. We found that pathogens modeled as a function of survival had a positive effect; turtles detected with a pathogen were two to six times more likely to survive than those without detected pathogens. However, this may be an artifact of high, unbiased pathogen prevalence paired with a relatively low probability of pathogen detection via intermittent testing. This analysis provides important estimates of apparent survival for the declining eastern box turtle and valuable information on the interaction between pathogen detection and estimates of individual survival, which can be used to better understand the drivers of population persistence in this species.

ABSTRACTMaynard Smith's proposed two‐fold cost of sex states that one of the disadvantages of clonal reproduction is the decreased ability to persist in dynamic ecosystems. However, the long‐term persistence of some clonal alloploid lineages suggests that these lineages may not always be so ephemeral in nature. Understanding the stability of these lineages over time can inform our understanding of the advantages of an asexual mode of life. Here we investigate a gynogenetic allopolyploid triploid, the Silvery Salamander (LJJ—historically referred to as Ambystoma platineum), an asexual lineage closely related to A. barbouri. However, in our focal populations, neither parental species is present, and another sexual host, A. texanum (TT), is a required sperm donor. Gynogenetic reproduction involving LJJ and its host A. texanum should result in a clone of the mother (LJJ); however, with the occurrence of kleptogenesis, it often can result in tetraploid hybrids (LJJT). LJJ is considered endangered in this population due to its restricted range coupled with concerns that the cryptic tetraploid (LJJT) could completely replace LJJ. Here we assess the level of LJJ × A. texanum hybridization in nine ephemeral wetland populations in east central Illinois. Using species‐specific microsatellite loci, we compared the prevalence of LJJ and LJJT genotypes across localities and years. We find variation across ponds and developmental stages but suggest relative stability over time. Given the considerable amount of environmental degradation and loss, we suggest continued monitoring of this unique segment of biodiversity to ensure its persistence into the future.

A primary response to the Clean Water Act has been the development of numerous indices of biotic integrity (IBI) using various taxonomic groups to assess the condition of water bodies in the United States. The IBI is a frequently used approach for assessing the ecological integrity of streams; however, it is less commonly applied to wetland systems, despite the scientific and policy needs to assess wetland condition and develop ecological performance goals for wetland monitoring, creation, and restoration. Although a great deal of attention has been given to wetland acreage due to the "no net loss" policy over the past two decades, much less attention has been given to the ecological integrity of these same wetlands. To assess wetland health and function, regulatory agencies and land managers need clear performance standards that are based on the ecology of wetland organisms. To address this need, we sampled 242 wetlands distributed across 45 managed properties in Illinois, U. S.A. over three years (2012-2014) for adult and larval amphibians (12 samples per wetland). Amphibian assemblages make up critical ecological components of many wetland ecosystems and are considered useful bioindicators of environmental change because they are sensitive to various forms of environmental and habitat alteration. We developed an amphibian-based IBI (hereafter aIBI) for the assessment of seasonal wetlands and optimal sampling strategies to best quantify these wetlands (i.e., assessment of time and effort). The aIBI applied a conservation coefficient to all species captured; conservation coefficients were developed through a survey of amphibian experts (n = 16). We captured 25 amphibian species with an average of 7.11 amphibian species (range 1-13) per site. Using the species occurrence data collected from both dipnet and minnow trap surveys over 12 survey periods, we assessed the average percent error in the estimation of aIBIs and species richness. Our results show that a wetland can be accurately evaluated with amphibians in one field season, but that multiple samples during that field season will be necessary. We show that 4-5 surveys using multiple methods distributed throughout the spring and summer are required. This level of effort may be incongruent with existing definitions of 'rapid assessment', but it is the necessary level of effort needed to confidently and accurately evaluate wetlands using amphibians as an indicator taxon. Additionally, aIBI provides a more robust framework to assess wetland integrity compared to simple species richness or other similar methodologies.

Abstract Snakes are difficult to study due to their cryptic coloration, minimal movements, and use of inaccessible habitats. Although well‐timed surveys during a species' active season can result in higher detection rates and conserve survey resources (i.e., time and money), survey effort may not ensure the detection of rare and cryptic species. Thus, in such instances, a strategic species‐specific sampling design is needed. The Kirtland's snake (Clonophis kirtlandii) is a rare, cryptic species assumed to be experiencing range‐wide declines. Naturalists have noted the disappearance of Kirtland's snakes from various habitats since the early 1970s. The primary objective of our study was to determine detection of Kirtland's snakes and the environmental and temporal factors influencing detection. We calculated the effort needed to detect individuals at sites by estimating detection probabilities of 3 known Kirtland's snake populations in Illinois from 2019 to 2021. Based on 77 Kirtland's snake detections over 226 site visits (34.1%) across 3 study sites, we found that high cloud cover, moderate air temperature, and low relative humidity enhanced the detection probability of this species. The middle of May to the beginning of July was the best time to conduct surveys when detection rates were highest. As our results suggested, it is imperative to establish strategic monitoring programs maximizing conservation resources to document populations for conservation action and range shifts for species of conservation concern, such as Kirtland's snakes.

As anthropogenic changes continue to ecologically stress wildlife, obtaining measures of gene flow and genetic diversity are crucial for evaluating population trends and considering management and conservation strategies for small, imperiled populations. In our study, we conducted a molecular assessment to expand on previous work to elucidate patterns of diversity and connectivity in the remaining disjunct Eastern Massasauga Rattlesnake (Sistrurus catenatus) hibernacula in Illinois. We assayed genetic data for 327 samples collected during 1999-2015 from the Carlyle Lake study area across 21 microsatellite loci. We found hibernacula formed distinct genetic clusters corresponding to the three main study areas (Dam Recreation Areas, Eldon Hazlet State Park, and South Shore State Park). Genetic structuring and low estimates of dispersal indicated that connectivity among these study areas is limited and each is demographically independent. Hibernacula exhibited moderate levels of heterozygosity (0.60-0.73), but estimates of effective population size (5.2-41.0) were low and track census sizes generated via long-term mark-recapture data. Hibernacula at Carlyle Lake, which represent the only Eastern Massasauga remaining in Illinois, are vulnerable to future loss of genetic diversity through lack of gene flow as well as demographic and environmental stochastic processes. Our work highlights the need to include population-level genetic data in recovery planning and suggests that recovery efforts should focus on managing the three major study areas as separate conservation units in order to preserve and maintain long-term adaptive potential of these populations. Specific management goals should include improving connectivity among hibernacula, maintaining existing wet grassland habitat, and minimizing anthropogenic sources of mortality caused by habitat management (e.g., mowing, prescribed fire) and recreational activities. Our molecular study provides additional details about demographic parameters and connectivity at Carlyle Lake that can be used to guide recovery of Eastern Massasauga in Illinois and throughout its range.

Objectives To identify associations between success following application for consultant physician posts and demographic factors. Design Logistic regression analysis of nationwide survey data. Setting United Kingdom (UK) physicians with a recent certificate of completion of training (CCT). Participants All UK trainee physicians who received a CCT between 2010 and 2019 were surveyed. Respondents were excluded if they had not applied for a consultant post or if application data were incomplete. Main outcome measures The primary outcome measure was success over the entire consultant application process, i.e. shortlisted and offered the post following the first application. Secondary outcomes were: shortlisted following first application and offered a consultant post at first interview. Results From 7037 CCT holders surveyed, 50.7% responded. While 1198 (59.7%) respondents were white, 760 (37.9%) were from minority ethnic groups and 50 (3.5%) were of unknown ethnicity. Primary medical qualification (PMQ) country was the UK in 75.3% (n = 1512). On multivariable logistic regression analysis the independent negative associations with success were: minority ethnicity (odds ratio [OR] 0.55, 95% confidence interval [CI] 0.43-0.71); p < 0.001) vs. white; PMQ from Europe (OR 0.47, 95% CI 0.28-0.79; p = 0.004) or Asia (OR 0.68, 95% CI 0.49-0.96; p = 0.027) vs. UK PMQ; year of CCT 2012 (OR 0.40, 95% CI 0.24-0.68; p = 0.001), 2013 (OR 0.39, 95% CI 0.23-0.65; p < 0.001), and 2014 (OR 0.26, 95% CI 0.15-0.43; p < 0.001) vs. 2019. Specialties associated with lower success rates included Cardiology, Endocrinology, Genitourinary medicine, Palliative care, Renal and Respiratory, compared to Acute medicine. Conclusions Minority ethnic group candidates for consultant physician posts had lower success rates compared to white candidates after correction for important variables including specialty, time from and country of PMQ. This finding requires further evaluation to identify the causes for this variation.

Introduction British Society of Gastroenterology (BSG) Workforce reports have identified widespread consultant vacancies, excessive workloads and insufficient training posts to meet current and future service demand. The COVID-19 pandemic has put further strain on an already overstretched and often demoralised workforce. Methods The Medical Workforce Unit of the Royal College of Physicians (RCP) undertakes an annual census of consultants each October. Data for gastroenterologists and hepatologists from the 2019 census, January 2020 RCP consultant wellbeing survey and the General Medicine Council (GMC) between 2012 and 2020 were examined. Results The number of UK licensed gastroenterologists increased from 1,703 to 2,013 from 2014 to 2019. However, in recent years, fewer gastroenterologists have joined the GMC register annually (151 in 2014 and 115 in 2018). Consequently, 45% of advertised consultant posts in gastroenterology were unfilled and 8% of all posts filled by a locum in 2019, mostly due to a lack of applicants, with demand outstripping supply. There has also been a smaller increase (9% versus 14%) in gastroenterology trainee numbers compared to other medical specialties between 2012 and 2020. The average number of population per full time equivalent gastroenterologist in the UK is 43,913 but there are significant regional variations with some regions much more poorly served (Thames Valley 63,456, North Wales 59,444, Kent, Surrey and Sussex 50.974, and East of England 50,487). Gastroenterologists work a mean of 1PA unpaid in addition to contracted PAs. This leads to gastroenterologists saying they always, or most of the time in 59% work excessive hours and in 61% have an excessive workload. Gastroenterology is the major specialty at highest risk of burnout with lower (worse) mean mental wellbeing scores. Consequently, 51% of gastroenterologists reported that work affected their relationship with their partner and 53% with their children in the past year. 52% described their morale in 2019 as being worse than in 2018. 18% of gastroenterologists reported feeling bullied or harassment over the past year by managers or fellow consultants. Despite these issues, 87% of gastroenterologists reported they were always or often satisfied with specialty work, but only 21% with general internal medicine work. Conclusions Unfilled consultant vacancies, high workloads, low wellbeing scores and morale were evident in gastroenterology and hepatology even before the COVID-19 pandemic. Gastroenterology is the major medical specialty at highest risk of burnout. At a time when the NHS workforce is under increasing pressure, these issues must be urgently addressed to improve the working lives of gastroenterologists and hepatologists.

INTRODUCTION:21% of NHS staff are from Black, Asian and minority ethnic (BAME) backgrounds yet account for a disproportionately high number of medical-staff deaths from COVID-19. Using data from the published OpenSAFELY Collaborative, we analysed consultant physicians to determine those at increased risk of COVID-19 related death.METHODS:Data from 13,500 consultant physicians collected by the Royal College of Physicians were analysed to determine those at an increased risk of death from COVID-19, assuming no comorbidities.RESULTS:The data reveal a picture in which a third of consultant physicians have a hazard ratio (HR) >1 for dying from COVID-19; one in five have HR >2; one in 11, HR >3; and one in 40, HR >4. Of concern are the risks to male physicians aged ≥60 with HR >3.8. Sub-specialties including cardiology, endocrine and diabetes, gastroenterology, haematology, neurology and rheumatology have a greater risk profile due to high proportion of men, physicians of older age, and proportion of BAME individuals.CONCLUSION:A third of consultant physicians have an increased risk of a COVID-19-related death, and one in five have a higher relative risk (HR >2). The risk is mainly driven by age, gender, and ethnicity, the risk is highest in male consultant physicians over 60, especially from BAME backgrounds.

Model-based approaches that attempt to delimit species are hampered by computational limitations as well as the unfortunate tendency by users to disregard algorithmic assumptions. Alternatives are clearly needed, and machine-learning (M-L) is attractive in this regard as it functions without the need to explicitly define a species concept. Unfortunately, its performance will vary according to which (of several) bioinformatic parameters are invoked. Herein, we gauge the effectiveness of M-L-based species-delimitation algorithms by parsing 64 variably-filtered versions of a ddRAD-derived SNP data set collected from North American box turtles (Terrapene spp.). Our filtering strategies included: (i) minor allele frequencies (MAF) of 5%, 3%, 1%, and 0% (= none), and (ii) maximum missing data per-individual/per-population at 25%, 50%, 75%, and 100% (= no filtering). We found that species-delimitation via unsupervised M-L impacted the signal-to-noise ratio in our data, as well as the discordance among resolved clades. The latter may also reflect biogeographic history, gene flow, incomplete lineage sorting, or combinations thereof (as corroborated from previously observed patterns of differential introgression). Our results substantiate M-L as a viable species-delimitation method, but also demonstrate how commonly observed patterns of phylogenetic discordance can seriously impact M-L-classification.

Studies to assess wildlife health commonly evaluate clinical pathology changes, immune responses, pathogen presence, and contaminant exposure, but novel modalities are needed to characterize the unique physiologic responses of reptiles. Lactate is an indicator of hypoperfusion and/or anaerobic respiration and can be quickly and easily measured using a point-of-care analyzer. This study evaluated baseline blood lactate concentrations in free-living eastern box turtles (Terrapene carolina carolina, n = 116) using a point of care analyzer and then determined the effect of handling time, physical examination (PE) abnormalities, and quantitative polymerase chain reaction pathogen detection (Terrapene herpesvirus 1, Mycoplasma sp., Terrapene adenovirus) on lactate concentrations. Blood lactate concentrations were higher in turtles with Terrapene herpesvirus 1 (n = 11), quiet mentation, and increased packed cell volume (P < 0.05). Lactate concentrations increased between initial capture and PE, with peak values reaching 129 min after capture. Lactate at PE was positively associated with baseline lactate concentrations. Turtles with Terrapene herpesvirus 1 may have alterations in blood flow, oxygen delivery, or activity patterns, driving increases in baseline lactate. Increased handling time likely leads to more escape behaviors and/or breath holding, causing turtles to undergo anaerobic metabolism and raising lactate concentrations. Overall, lactate measured by a point of care analyzer shows variability caused by capture and health factors in eastern box turtles and may be a useful adjunctive diagnostic test in this species after full methodologic validation.

Environmental DNA represents a potentially cost-effective method for detection of rare and cryptic species, for which traditional sampling can be logistically costly. We tested species-specific probe-based assays to detect eDNA for the federally threatened eastern massasauga rattlesnake and the causative agent of snake fungal disease, Ophidiomyces ophiodiicola . We collected water samples from 100 crayfish burrows (over-wintering refugia) during spring emergence and screened them for snake and fungal DNA. We detected snake DNA in two samples and failed to detect O. ophiodiicola DNA. Given the high density of both target species at the sites, we suggest these methods may not offer a logistical and financial improvement over traditional sampling protocols.

Hybridization occurs differentially across the genome in a balancing act between selection and migration. With the unprecedented resolution of contemporary sequencing technologies, selection and migration can now be effectively quantified such that researchers can identify genetic elements involved in introgression. Furthermore, genomic patterns can now be associated with ecologically relevant phenotypes, given availability of annotated reference genomes. We do so in North American box turtles (Terrapene) by deciphering how selection affects hybrid zones at the interface of species boundaries and identifying genetic regions potentially under selection that may relate to thermal adaptations. Such genes may impact physiological pathways involved in temperature-dependent sex determination, immune system functioning, and hypoxia tolerance. We contrasted these patterns across inter- and intra-specific hybrid zones that differ temporally and biogeographically. We demonstrate hybridization is broadly apparent in Terrapene, but with observed genomic cline patterns corresponding to species boundaries at loci potentially associated with thermal adaptation. These loci display signatures of directional introgression within intra-specific boundaries, despite a genome-wide selective trend against intergrades. In contrast, outlier loci for inter-specific comparisons exhibited evidence of being under selection against hybrids. Importantly, adaptations coinciding with species boundaries in Terrapene overlap with climatic boundaries and highlight the vulnerability of these terrestrial ectotherms to anthropogenic pressures.

Wildlife disease surveillance and pathogen detection are fundamental for conservation, population sustainability, and public health. Detection of pathogens in snakes is often overlooked despite their essential roles as both predators and prey within their communities. Ophidiomycosis (formerly referred to as Snake Fungal Disease, SFD), an emergent disease on the North American landscape caused by the fungusOphidiomyces ophiodiicola, poses a threat to snake population health and stability. We tested 657 individual snakes representing 58 species in 31 states from 56 military bases in the continental US and Puerto Rico forO.ophiodiicola.Ophidiomyces ophiodiicolaDNA was detected in samples from 113 snakes for a prevalence of 17.2% (95% CI: 14.4-20.3%), representing 25 species from 19 states/territories, including the first reports of the pathogen in snakes in Idaho, Oklahoma, and Puerto Rico. Most animals were ophidiomycosis negative (n = 462), withOphidiomycesdetected by qPCR (n = 64), possible ophidiomycosis (n = 82), and apparent ophidiomycosis (n = 49) occurring less frequently. Adults had 2.38 times greater odds than juveniles of being diagnosed with ophidiomycosis. Snakes from Georgia, Massachusetts, Pennsylvania, and Virginia all had greater odds of ophidiomycosis diagnosis, while snakes from Idaho were less likely to be diagnosed with ophidiomycosis. The results of this survey indicate that this pathogen is endemic in the eastern US and identified new sites that could represent emergence or improved detection of endemic sites. The direct mortality of snakes with ophidiomycosis is unknown from this study, but the presence of numerous individuals with clinical disease warrants further investigation and possible conservation action.

BACKGROUND:To the authors' knowledge, this is the first report of mast cell neoplasia in a koala (Phascolarctos cinereus).CASE REPORT:An adult female koala was presented for rapidly deteriorating health and death of a pouch young. Significant weight loss was apparent despite supplemental feeding; the abdomen was distended; and the koala was weak and mentally depressed. Haematology revealed a significant mastocytosis with a concurrent population of atypical mononuclear cells. The koala was euthanised and tissues were collected for histology. Bone marrow, lymph node, lung, stomach and spleen exhibited significant infiltration by mast cells. Atypical round cells consistent with those identified in the peripheral blood were also identified in the marrow. A diagnosis of systemic mastocytosis and probable mast cell leukaemia was made. Immunocytochemical and immunohistochemical staining was not able to further characterise the atypical cell population, and the mast cells exhibited only weak staining with CD117.CONCLUSION:The histological diagnosis, in this case, was systemic mastocytosis and myeloproliferative disease of uncertain origin. There was a dominant population of mast cells in the peripheral blood and marrow, and a population of circulating atypical mononuclear cells, appearing similar to mast cell leukaemia-acute myeloid leukaemia in humans.

Population surveying and monitoring are important for identifying conservation needs and tracking trends in populations, communities, and ecosystems over time and laying the groundwork for conservation management and policy decisions. If species or populations go undetected because of inadequate effort or sampling design, protection and management cannot be properly provided. Due to the widespread loss of populations, the Eastern Massasauga (a rattlesnake) was recently listed as a federally threatened species in the United States; it is also listed as threatened in Canada. Given its current conservation status, there is considerable interest at state and federal levels in determining how to best survey for Eastern Massasaugas to aid in management decisions. Using a 16-year dataset, we examined the relationships among environmental, temporal, area, management, and search effort factors on the detection probability of Eastern Massasaugas. We found that four abiotic parameters (solar irradiance, shaded air temperature, three-day maximum air temperature, and humidity) and three search parameters (effort per researcher, search area, and search time of day) influenced detection of Eastern Massasaugas. As the current biodiversity crisis continues, the cost-effective use of resources and scientific expertise will continue to increase in importance. We hope our results stimulate similar analyses in other taxa, which will be critical for designing and implementing regional survey and monitoring programs.

The acute phase response is a highly conserved reaction to infection, inflammation, trauma, stress, and neoplasia. Acute phase assays are useful for wildlife health assessment, however, they are infrequently utilized in reptiles. This study evaluated erythrocyte sedimentation rate (ESR) in eastern (Terrapene carolina carolina) and ornate box turtles (Terrapene ornata ornata) and hemoglobin-binding protein (HBP) in T. ornata. Erythrocyte sedimentation rate in 90 T. carolina and 105 T. ornata was negatively associated with packed cell volume and was greater in unhealthy turtles (p < 0.05). Female T. ornata had higher ESR values than males (p < 0.05). Measurement of ESR with a microhematocrit tube proportionally overestimated values from a commercial kit (Winpette), though both methods may retain utility with separate reference intervals. Hemoglobin-binding protein concentration in 184 T. ornata was significantly increased in adults and unhealthy turtles (p < 0.05). Erythrocyte sedimentation rate values were similar between seasons and populations, and HBP values were consistent between years, indicating that these analytes may have more stable baseline values than traditional health metrics in reptiles. This study demonstrates that ESR and HBP are promising diagnostics for health assessment in wild box turtles. Incorporating these tests into wild herptile health assessment protocols may support conservation efforts and improve ecosystem health monitoring.

We tracked yearly variation in breeding phenology in relation to weather parameters in a common European toad population from the southeastern part of its range. Phonological data were collected from 2001 to 2003 and from 2011 to 2017 and compared to open-access daily weather data from a nearby weather station. Data analysis revealed no significant effect of weather on initiation of breeding and a negative relationship between mean daily humidity and mean cloud cover on the duration of breeding season (P = 0.03). Further analysis showed a decreasing trend in breeding season humidity in the past 70 years. Our results predict a tendency toward longer toad breeding seasons in years with drier winter/spring. A projected decrease in humidity in this region could prolong toad breeding season, potentially exposing adults to higher predation. Therefore, the scenario of further decline in our study population should be considered and conservation measures planned accordingly.

Physicians with training in point-of-care ultrasound (POCUS) can reliably estimate left ventricular ejection fracture (LVEF). However, those without experience or at novice level may misestimate LVEF. The reference standard for calculation of LVEF involves obtaining both apical four-chamber (AP4) and apical two-chamber (AP2) views, and tracing the endocardial border. In POCUS, these views are difficult to obtain due to a number of patient, machine and environmental factors. Physicians may therefore use other views such as the parasternal long axis (PLAX) when determining LVEF, and do not assign specific quantitative values to LVEF.