Peer Community Journal

The detection and monitoring of bat species using non-invasive sampling and molecular techniques has become increasingly popular in recent years. In Europe, these approaches have been applied to identify horseshoe bats of the genus Rhinolophus, which includes five species: R. hipposideros, R. ferrumequinum, R. euryale, R. mehelyi and R. blasii. While species-specific real-time PCR assays exist for R. ferrumequinum and R. hipposideros, no unified panel of real-time PCR assays currently enables the identification of all five European Rhinolophus species from non-invasively collected samples. Here, we developed five species-specific real-time PCR assays, each targeting interspecies nucleotide variation within the mitochondrial cytochrome b gene. To enhance single-base discrimination, RNase H-dependent PCR (rhPCR) primers were employed, incorporating cleavable blocked primers that require perfect complementarity for extension. The assays were applied to droppings non-invasively collected from 18 caves and one church in Serbia and Romania. Of the 149 samples analysed, 131 (88%) yielded successful amplification of Rhinolophus DNA. Detection probabilities for the three species identified in the field ranged from 0.49 to 0.82. Occupancy estimates varied, with R. euryale showing the highest (0.86; UI: 0.69-0.97) and R. mehelyi the lowest (0.23; UI: 0.08-0.43). The assays were capable of detecting up to three species concurrently within a single pooled sample (approximately 15 droppings). These assays are especially valuable for detecting R. mehelyi, given its rarity and uncertain distribution, and offer a robust tool for monitoring Rhinolophus populations across Europe.

Mark-recapture analyses on the delineation of natural populations between areas often assume random sampling, with a between/within (B/W) area resighting ratio that declines towards zero as the population components of two areas become more-and-more isolated from one another, with fewer-and-fewer individuals mixing between areas. I use an individual based population model split in two areas to simulate this result, analysing also for the potential effects of the space-time fidelity of the mark-recapture sampling in the areas. I find that small B/W resighting ratios--that traditionally is taken as evidence of population isolation--can easily be observed within a completely mixing population if a random sampling scheme is restricted in space and/or time. Random sampling within restricted areas and time windows is not sufficient to estimate mixing rates and population isolation between areas, unless the resighting rates are analysed by a method that accounts both for the space-time fidelity of the scientific sampling scheme and the space-time fidelity of the distributional behaviour of the individuals in the population.

Biological sequences are known to be not random. Thus, the comparison of in silico restriction fragment distributions of random and biological sequences may be an indicator of this non-randomness. Our analyses show that for most of the tested combinations of restriction enzyme and genome sequence the fragments per Megabase of the biological sequence deviate at least more then 10% from the corresponding random sequence. This deviation goes into both directions, i.e. clearly increased values are as common as clearly decreased values. Although there is no species- or restriction-enzyme-specific effect, a clear impact of the GC content both of the restriction site and of the genome sequence can be seen. In contrast to the random sequences, the genome sequences show distinct peaks in their fragment length distributions, hinting to repetitive elements such as transposons.

The marbled teal (Marmaronetta angustirostris) is a widely distributed but declining waterfowl species, classified as Near Threatened globally and Critically Endangered in Spain. Despite ongoing conservation actions, including ex situ management and population reinforcement programmes, the genomic consequences of long-term captivity, inbreeding, and patterns of functional genetic variation remain unknown due to the absence of a species-specific reference genome. Here, we present the first chromosome-level genome assembly for this species. The genome was generated using PacBio HiFi long reads and Omni-C data, yielding a 1.15Gb assembly with a scaffold N50 of 76.95Mb. A total of 97.16% of the assembly was anchored into 36 chromosome-scale scaffolds, including the Z and W sex chromosomes. BUSCO analysis recovered 99.2% of conserved avian genes. Gene prediction was performed using both ab initio and homology-based strategies, resulting in 16,048 protein-coding genes. This resource provides a foundation for genomewide analyses of inbreeding, demographic history, and adaptive variation, and will support evidencebased in situ and ex situ conservation strategies for this threatened species.

Nature-Based Solutions are increasingly promoted to address current urban challenges. While their potential effects on vector-borne disease risks have been documented, data on Aedes albopictus, a known arbovirus vector, remain limited in France. A previous study showed that urban vegetation moderately increases the abundance of adult mosquitoes of this species, but the monitoring period lasted only six months. Using ovitraps, we monitored Ae. albopictus egg density dynamics over multiple years (2022 to 2024) and analysed its environmental predictors in various urban environments. We included lagged meteorological variables, land cover metrics, and the cumulated egg densities recorded in the previous weeks as environmental predictors. Both parametric (GLMM) and non-parametric (Random Forest) models were fitted to weekly egg counts per trap. Our findings highlight that (i) egg density dynamics were related to how vegetation classes structured the landscape, (ii) growing degree days and cumulated number of eggs recorded in specific lagged time windows were the main contributors to egg density, and (iii) the non-parametric and parametric models performed similarly in terms of prediction accuracy.

According to sexual selection theory, males should benefit more from mating with multiple partners than females do, as male investment into offspring production is typically lower. For females, empirical evidence indeed often shows diminishing returns or even costs of mating multiply. For males, the assumption often seems to be "the more, the better" - i.e., a steady increase of male reproductive success with mate number - but experimental tests of it are rare. Here we used a laboratory experiment with Trinidadian guppies (Poecilia reticulata), known for being promiscuous, to assess how pairing males weekly with 4 vs. 7 females affects both sexes reproductive performance (n = 32 polygynous males and 170 monogamous females). Increased polygyny delayed females reproductive onset by 9% and tripled their risk of reproductive failure. High-polygyny males fathered offspring with 49% more females and had 73% higher daily reproductive output. Yet, they needed 19% longer to initiate pregnancy, and only accumulated more offspring than low-polygyny males after two months. This study suggests that male mating performance is not unlimited. Especially when high extrinsic mortality selects for fast reproduction, less polygyny might be advantageous, and the strength of sexual selection perhaps more similar between the sexes than often assumed.

Alleles with opposing effects on fitness characters are said to exhibit selectional antagonistic pleiotropy (broadly construed so that effects are not necessarily confined to the same individual). A number of theoretical investigations considered the case where a pair of alleles at a locus influences two fitness components and derived the conditions giving rise to stable polymorphism under various assumptions about the mode of trait-interaction. Strikingly, many of these analyses concluded that the potential for maintaining polymorphism is strongly constrained by the joint influence of two factors: (1) the prevalence of weak selection coefficients over coefficients of large magnitude, and (2) the absence of beneficial dominance reversals (where the deleterious effects of each allele are partially or completely masked in the heterozygous genotype). Consequently, the conclusion that selective polymorphism is unlikely to be maintained by intralocus mechanisms of antagonistic pleiotropy has achieved widespread acceptance. Here we argue that such conclusions do not apply to any of the following models of antagonism: (i) additive trait-interaction, (ii) multiplicative trait-interaction, (iii) bivoltine selection, (iv) soft selection, (v) hard selection, and (vi) sexual antagonism. We demonstrate that the parameter space giving rise to stable allelic variation is quite large throughout, and moreover, the plenitude of suitable parameters neither depends on the strength of selection nor requires dominance reversal. Dominance coefficients associated with stringent conditions for stable polymorphism are shown to be atypical as compared to all feasible parameters, and best regarded as an outcome of adherence to a special relation: dominance with a constant magnitude and direction, which includes the case of additive allelic effects at a locus. Properties of single-locus equilibria (heterozygosity, allele frequency differentiation) are investigated, as well as the contribution of dominance schemes to the genetic variance in fitness characters in populations at multilocus linkage equilibrium. Author summaryAllelic variants at a locus with opposing effects on multiple fitness components (antagonistic fitness pleiotropy) have long been appreciated as a possible source of balancing selection. The prevalence of polymorphism owing to this form of natural selection, however, has been doubted on theoretical grounds due to the fact that standard assumptions of genetic models (namely, constant magnitudes for the dominance coefficients) are hardly conducive to the maintenance of polymorphism. The major exception to this conclusion lies with schemes that exhibit dominance reversal (where the direction of dominance for antagonistic alleles flips across fitness components). Here we conduct a geometric analysis of the space of polymorphism-promoting dominance parameters and conclude that the conditions for maintaining balanced alleles is unrestrictive, with non-reversals playing an underappreciated role.

The choice of appropriate methods to detect species is crucial for biodiversity monitoring. Camera trapping is currently one of the most widely used methods for characterizing mammal communities, although it requires substantial investment in equipment and personnel. In contrast, questionnaires administered to local populations provide a faster and more cost-effective alternative for assessing community composition, but may be influenced by respondent-related biases that compromise data reliability. This study evaluates the concordance between these approaches for characterizing the carnivore community in the Sierra de Segura (Jaen, southern Spain), using Cohens kappa coefficient, while also examining the individual and social factors shaping Local Ecological Knowledge (LEK). We deployed 24 camera-trap stations (144 trap nights) across a 25 km2 area to record carnivore presence. In parallel, we conducted two types of surveys with local residents (n = 103): (i) free-listing and (ii) image-based species recognition, while recording individual and social characteristics of respondents. Free-listing surveys tended to underreport species, whereas image-based surveys showed higher agreement with camera-trap data, although occasionally overestimating species presence. Higher concordance was associated with social factors indicative of closer and prolonged contact with the environment, such as permanent residence and ownership of agricultural land. Mammal communities differed between methods; however, agreement improved when respondents had higher LEK, while species-specific behavioral traits could also influence perception. Our findings demonstrate that image-based questionnaires can provide results comparable to camera trapping when respondents have strong connections to their natural surroundings. These results highlight the importance of both survey design and respondent selection in improving the accuracy of biodiversity monitoring, offering a transferable framework for integrating LEK into conservation protocols across diverse ecosystems. O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=126 SRC="FIGDIR/small/720805v1_ufig1.gif" ALT="Figure 1"> View larger version (35K): org.highwire.dtl.DTLVardef@d55c34org.highwire.dtl.DTLVardef@1985c66org.highwire.dtl.DTLVardef@1da576aorg.highwire.dtl.DTLVardef@1a10ccb_HPS_FORMAT_FIGEXP M_FIG C_FIG

Objectives: To determine the impact of discharge home on physical activity and sedentary behaviour following orthopaedic trauma. Design: Observational study. Setting: Acute hospital. Participants: Between October 2022 and January 2024, 31 adult orthopaedic trauma patients were recruited during hospital admission. Participants had either an isolated hip fracture or multi-trauma (i.e., a lower limb fracture, with an upper limb and/or spinal fracture). Interventions: Participants wore two activity monitors (activPAL3 and ActiGraphGT3x) during the final days of an acute hospital admission and the first five days at home. An interrupted time series analysis evaluated changes physical activity variables during the hospital to home transition. Participants were analysed individually using mixed-effects linear regression allowing the intercept to vary by participant. Main outcome measures: Primary outcome was daily steps; secondary outcomes included sedentary time and other activity measures. Results: Daily steps (mean +- SD) were higher at home (4552.4 +- 2639.5) compared to hospital (2597.8 +- 1450.8). Modelled results indicated a 27% increase in daily steps following hospital discharge (exp(beta946;): 1.27, 95% CI: 1.01,1.59, p=0.039) and a sustained improvement at home. No significant differences were observed between hip fracture and multi-trauma participants. Conclusion: Participants recovering from orthopaedic trauma showed a significant increase in daily step count upon discharge home from hospital, highlighting the positive impact of the home environment on activity levels. Further research is warranted to assess the effectiveness of interventions to improve activity levels in hospital (e.g., early intensive therapy) and at home (e.g., immediate home-based physiotherapy) in individuals following orthopaedic trauma.

Objective To examine the effects of combined oral contraceptive (OC) use on clinical markers of ovarian reserve by comparing Anti-Muellerian Hormone (AMH), antral follicle count (AFC), and ovarian volume (OV) before and after starting or stopping OC. Methods This analysis is based on data from a prospective cohort study conducted at the University Hospital Tubingen, Germany, as part of the IRTG-2804 project. A total of 54 healthy women were included and categorized into three groups based on their OC use status: OC starters (n = 12), stoppers (n = 16), and long-term OC-users (n = 26). Each participant underwent a transvaginal ultrasound (including AFC and OV) and serum sampling (including AMH) at two time points (S1 and S2), three to six months apart. OC starters were assessed first during the early follicular phase (day 1-7) and then during active OC intake (day 8-21), while stoppers were assessed in the reverse order. Long-term users were assessed twice during active OC intake. Results OC stoppers showed significant within-group increases in all ovarian reserve markers, including AMH ({Delta} = 2.57 ng/mL, p < .001), AFC ({Delta} = 3.88, p = .004), and OV, which almost doubled (1.94-fold increase; 95% CI [1.35, 2.80], p < .001). In contrast, OC starters exhibited a significant decline in AMH ({Delta} = -1.25 ng/mL, p = .013), but no changes in AFC or OV. No significant longitudinal changes were observed among long-term OC users. Conclusion AMH levels decrease after starting OC use whereas AFC and OV are not affected. In contrast, AMH, AFC, and OV recover within three to six months after stopping OC, suggesting a reversible suppression of ovarian reserve markers during OC use. These findings are clinically relevant for fertility counseling and for the interpretation of ovarian reserve markers in women using hormonal contraception.

ObjectiveUnlike several other fields of healthcare, little is known about the size of therapist effects on patient outcomes following rehabilitation for musculoskeletal conditions. We aimed to estimate the proportion of variance in patient outcomes from a structured rehabilitation program explained by therapist effects. MethodsFor our observational cohort study we accessed data from the national multicentre Good Life with osteoArthritis in Denmark (GLA:D) osteoarthritis management program. Analyses included 23,021 consecutive eligible adults with hip or knee osteoarthritis (mean (SD) age 65.0 (9.8) years, 71% female) treated by 657 therapists between October 2014 and February 2019. The primary outcome was [&ge;]30% reduction in pain intensity on 0-100 VAS at 3 months. Therapist effects were estimated as the variance partition coefficient (intra-class correlation coefficient (ICC)) from two-level random intercept logistic regression models before and after adjusting for patient-level case-mix factors and therapist-level characteristics (number of patients treated, days since therapist certification). Analyses were repeated for a range of secondary outcomes using multiply imputed data and complete-case analysis. Results52% of patients reported a [&ge;]30% reduction in pain intensity on 0-100 VAS at 3 months. In the null model the ICC was 0.007 (95%CI: 0.005, 0.009), which changed little after adjusting for patient- and therapist-level covariates. Upper confidence limits for ICC estimates across all secondary outcomes in multiply imputed and complete case analyses were less than 0.03. ConclusionsIn a nationally implemented osteoarthritis management program delivered by trained healthcare professionals, therapist effects made a minimal contribution to variation in patient outcomes. KEY MESSAGESO_ST_ABSWhat is already known on this topicC_ST_ABS Therapist effects - defined as the effect of a given therapist on patient outcomes as compared to another therapist - have been observed in several fields of healthcare and have important consequences for selection, training, and service improvement. In musculoskeletal rehabilitation five previous studies suggest that 1-12% of variation in patient-reported outcomes may be attributable to therapist effects, but these estimates were based on relatively small datasets resulting in substantial uncertainty. What this study addsOur cohort study analysed registry data from 2014-2019 on 23,021 patients and 647 trained therapists from the nationally implemented GLA:D structured osteoarthritis management program in Denmark. We found that therapist effects accounted for less than 3% of total variation in patient-reported pain and quality of life outcomes 3 months after beginning the program How this study might affect research, practice, or policyOur findings suggest that contextual factors that relate to therapist effects - therapist characteristics or therapist-patient interaction and alliance - make a minimal contribution to variation in patient outcomes from this structured, group-based rehabilitation intervention. Any contextual effects must be attributable to alternative sources, e.g. patient expectations, intervention setting.

The effects of the selection of livestock type (e.g., sheep or cattle) and grazing intensity on the soil seed bank of sand grasslands of conservation interest were studied. 25 grazed grassland sites classified into four grazing intensity categories were studied. The soil seed bank was analysed by seedling emergence; germinated seedlings were classified into morpho-functional, social behaviour type (SBT) and CSR strategy groups. The following hypotheses were tested: i) Diversity and density of soil seed banks are lower in sheep-grazed sites than in cattle-grazed ones. ii) The species composition, diversity, and density of the soil seed banks are more strongly affected by grazing intensity than by the livestock type. iii) Leaf traits, SBT and CSR strategy composition are highly affected both by livestock type and grazing intensity. The main effect of livestock type only affected seed bank density, while that of grazing intensity had a significant effect on most of the variables. Most of the studied variables were affected by the interaction of grazing intensity and livestock type. Total seed bank density was lower at all grazing intensity levels in sheep-grazed sites than in cattle-grazed ones, especially close to frequently visited places. We found that sheep grazing sustained a much lower total seed bank density and lower density of species of natural and semi-natural habitats regardless to the grazing intensity. Thus, livestock type must be carefully selected and high-intensity sheep grazing should be avoided in the long-run when managing sand grasslands. HighlightsO_LIThe soil seed banks of sheep and cattle grazed sand grassland were studied C_LIO_LIEffect of grazing intensity found the most important driver of seed bank diversity and density C_LIO_LIThe total soil seed bank density was higher in cattle than sheep grazed sites C_LIO_LIBoth intensity and livestock type must be considered in the grassland management planning C_LIO_LIHigh intensity sheep grazing should be avoided in sand grassland management C_LI

Wildlife rehabilitation plays a central role in the conservation of threatened primates, yet parasite dynamics during captivity are rarely reported, particularly in relation to release readiness. We investigated gastrointestinal helminth infection patterns in rehabilitating Javan slow lorises (Nycticebus javanicus), a Critically Endangered species heavily impacted by the illegal wildlife trade. Using repeated fecal sampling (147 samples from 19 adults) and Bayesian mixed-effects models, we examined parasite richness, Shannon diversity, infection probability, and egg-shedding intensity in relation to release readiness status, sex, housing condition, and time since anthelmintic treatment. Four nematode taxa identifiable through egg morphology were detected: Strongyloides spp., strongylids, oxyurids, and Trichuris spp.. Parasite richness and Shannon diversity showed no credible associations with release readiness or other host and management variables. In contrast, infection probability for Strongyloides spp. and strongylids increased with time since deworming, and Strongyloides egg counts exhibited a similar temporal pattern, consistent with post-treatment reinfection dynamics. Release readiness did not predict detection probability or parasite intensity for any parasite group, despite marked differences in captivity duration and health history between individuals deemed ready for release or not. These findings indicate that gastrointestinal helminth dynamics in rehabilitating slow lorises are driven primarily by treatment-related temporal processes and individual-level heterogeneity rather than coarse host classification. They also highlight the need for longitudinal parasite monitoring and for future work evaluating how infection dynamics, management interventions, and host health relate to rehabilitation and translocation outcomes..

In nature, the most common drivers of lethal aggression are predation and territorial defense. In northeastern Poland, the yellow-necked mouse (Apodemus flavicollis) coexists with several rodent species, including the bank vole (Clethrionomys glareolus). Compared to voles, A. flavicollis is larger, physically stronger, more aggressive, and dominant in the social ecosystem. However, no visually documented instance of a lethal attack by this species has been reported up to date. Here, we present the first recorded case of a fatal attack by a yellow-necked mouse following an encounter with a bank vole. A near-lethal attack is also reported. Importantly, these attacks were not predatory, as no consumption occurred. The attacks appeared instead to be related to interspecies competition, i.e., to competitive interactions between two species that live in the same habitat and use the same type of resources. Notably, while the aggressiveness of yellow-necked mice towards bank voles was known, it was unknown that it could take such extreme forms. Since, in rodents, most competition-related agonistic interactions are aimed at distancing the competitor, the physical destruction of the competitor appears as a surprisingly extreme way of addressing the game of interspecies competition through definitive removal of the opponent. Our observations highlight the need for further research on interspecific aggression among small mammals. They also emphasize the importance of field-based methods, such as camera trapping and continuous video monitoring, which allow for direct observation of animal behavior in natural settings and can reveal rare or previously overlooked interactions.

Accurate home range knowledge is essential for conserving species that are highly dependent on certain types of habitats. The critically endangered European mink (Mustela lutreola) is a wetland specialist whose movements are constrained by riparian and wetland habitats. In dendritic landscapes, conventional home range estimators such as Minimum Convex Polygons tend to include unsuitable habitats in estimated home ranges. Using VHF telemetry data from 16 individual-years tracked in France between 1996-1999 and 2020-2022, we compared four methods: Kernel Density Estimator (KDE), an adaptative sphere-of-influence local convex hull (a-LoCoH), a newly developed Ecological Home Range method (EHR), and a Generalized Additive Model (GAM) approach integrating hydrographic covariates. Our objective is to determine which method best accounts for the European minks specialization in wetlands, considering the spatial distribution of locations. Evaluation with wetland-specific metrics showed KDE consistently overestimated range extent and included unsuitable habitats, and a-LoCoH yielded mixed results, but these indicated that the method was not effective in excluding unused habitats. It was EHR and GAM methods that aligned more closely with ecological constraints. We therefore recommend GAM because it matches our objective and has the capacity to integrate additional environmental variables. Using the GAM, male home ranges averaged 3,074 ha--26 times larger than female ranges (116 ha)-- and were significantly larger in river than marsh landscapes. These are the largest ranges reported for the species. Large spatial requirements heighten vulnerability to road fatality and predation, both significant threats for remaining French populations. Our findings highlight the need for conservation strategies that integrate precise, habitat-based range estimates. The GAM method offers a robust, adaptable framework for managing European mink and other semi-aquatic species in complex landscapes.

Sleeping and feeding are crucial for survival of any animal. In case of large predators, knowing where these activities occur can help us understand their behavioural adaptations for coexisting with people and could help mitigating human-carnivore conflicts. Leopard (Panthera pardus) is an elusive and highly adaptable large felid that mostly lives outside protected areas and can survive also in close proximity to humans. However, most leopard research in Africa has been conducted in protected areas and we poorly understand leopards habitat selection while resting and hunting. To shed light on their coexistence with humans, we investigated habitat features influencing leopard selection of resting and kill sites on farmlands in central Namibia, using generalized linear mixed models (GLMM) under a use-availability study design and blinded field-sampling. Leopards primarily selected resting sites that were located in mountainous, steep, rugged terrain and sites with good concealment while kill sites were selected in mountainous habitats. Human infrastructure did not affect leopard resting and kill site selection. Thus, the capacity of leopards to perform essential life-supporting behaviours while coexisting with people appears to be primarily driven by their ability to remain concealed, rather than spatially avoiding humans.

Widow spiders of the genus Latrodectus are important animals for biomedical, pest and conservation research. Here, we present the assembled genomes of two closely related Latrodectus species: the Australian L. hasselti and the New Zealand endemic L. katipo. The genome of L. katipo consists of 13 scaffolds likely corresponding to chromosomes (90% of the total length) and 1267 short scaffolds (10%). It has a total length of 1.5 Gbp and BUSCO of 94.9%. The genome of L. hasselti consists of 379 scaffolds and has a total length of 1.7 Gbp and a BUSCO score of 95.4%. The repeat content is very similar in both genomes with a total proportion of 37.2% for L. katipo and 39.9% for L. hasselti. Genome annotation predicted 12706 and 15111 genes for L. katipo and L. hasselti respectively. An ortholog analysis shows large overlap between orthogroups suggesting either duplication events in L. hasselti or loss of genes in L. katipo.

Semi-aquatic mammals lie at the intersection of several key conservation issues such as wetland deterioration or species invasions, and monitoring their distribution in space and time is essential to inform conservation strategies. However, gathering information about their presence is challenging due to their elusive lifestyle and generally low abundance. The Eurasian otter (Lutra lutra), a near-threatened and strictly protected species in Europe, is currently recolonizing part of its historical range. Its high conservation interest, combined with a dynamic more commonly associated with range-expanding or invasive species, makes it a particularly compelling case study. Otter monitoring has traditionally relied on scat surveys, but recent environmental DNA (eDNA) and camera-trapping initiatives have emerged offering promising complementary tools. Yet, these approaches have rarely been formally compared, either to one another or across regions. Here, we compared the efficiency of spraint surveys, camera traps, and eDNA for detecting otters, and assessed how their performance varied among four catchments in southern France where the species is known to be present. All three methods provided otter detections with varying efficiency. Scat surveys were the most effective method, with an average detection probability of 0.71 and no strong variability between catchments. Although camera-traps had the lowest detection rate, they provided detections at two of the four sites where no spraint was found, highlighting the complementarity of these two approaches. Detection rates varied greatly between individual cameras rather than between catchments, underscoring sensitivity to camera-placement. eDNA showed important variability between catchments, with detection probabilities differing by roughly sixfold across regions. All in all, our results highlight differences in efficiency between methods and across environmental conditions, and show the value of combining approaches for future monitoring programs.

Mycoplasmopsis [Mycoplasma] agassizii is one of the principal pathogens associated with upper respiratory tract disease (URTD) in tortoises, yet its epidemiology in European wild chelonian populations remains poorly understood. The pathogen has been linked to population declines in some wild tortoise populations and is frequently detected in captive tortoises, where infections may persist subclinically and prolonged contact can facilitate transmission. In this context, the pet trade and the release or escape of captive individuals represent potential pathways for pathogen exchange between captive and wild populations. We assessed the presence and prevalence of M. agassizii in wild Mediterranean tortoises in Spain and compared infection patterns with captive populations. A total of 259 tortoises were sampled between 2020 and 2025, including spur thighed tortoises (Testudo graeca; 127 wild; 63 captive) and Hermanns tortoises (Testudo hermanni; 46 wild; 23 captive). Detection of M. agassizii was performed using PCR. The pathogen was detected in both species, but prevalence patterns differed markedly between captivity status and species. High prevalence was consistently observed in captive individuals of both species. In contrast, wild populations showed species-specific patterns: T. graeca exhibited very low or absent prevalence across wild populations, whereas T. hermanni showed comparatively higher prevalence in the wild. These results provide the first baseline assessment of M. agassizii occurrence in Mediterranean tortoises in Spain and highlight the importance of incorporating pathogen surveillance into conservation and management strategies for European chelonian populations.

Parasite infections in wildlife vary across time and space due to interactions among host biology, ecological processes, and climatic variability. Under ongoing climate change, understanding how temperature, precipitation, or humidity influences parasite dynamics is important for predicting shifts in infection patterns and host-parasite interactions. Here, we examine how variation in climatic conditions is associated with helminth infections in a free-living rodent, the bank vole (Myodes glareolus), across 17 years and multiple spatial scales. Using zero-inflated generalised linear models, we quantified the effects of climatic variables on individual parasite burden. Climatic conditions (temperature and humidity or precipitation) affected helminth infections across all analysed scales, though the strength and direction of these effects differed among parasite species and between temporal and spatial scales. In the temporal dataset, parasite load was associated with seasonal variation in weather conditions, whereas in the spatial datasets, infection levels were linked to yearly average climatic variables. The differences reflect species-specific parasites life histories and transmission strategies. Our findings highlight the importance of analysing individual parasite species rather than overall parasite load or aggregated infection indices when assessing the impacts of climatic variation on host-parasite dynamics.