Over the last few decades North American flying squirrels (Glaucomys spp.) have experienced dramatic northward range shifts. Previous studies have focused on the potential effects of warming winter temperatures, yet the hypothesis that rising summer temperature had a role in these range shifts remained unexplored. We therefore sought to determine the effect of high environmental temperatures on the thermoregulation and energetics of flying squirrels in an area of the Northeast of North America with a recent species turnover. Unable to find a logistically feasible population of the northern species (Glaucomys sabrinus), we focused on Southern Flying Squirrels (G. volans). Using flow-through respirometry, we measured the relationship between metabolic rate, evaporative water loss, and body temperature at high ambient temperatures. We also measured core body temperature in free-ranging flying squirrels using temperature-sensitive data loggers. We detected no significant increase in metabolic rate up to ambient temperatures as high as 40 °C. However, evaporative water loss increased at temperatures above 36.2 °C. Free-ranging body temperature of flying squirrels followed a circadian pattern with a ~2 °C difference between active and resting phase modal body temperatures. Rest-phase body temperatures were influenced by environmental temperatures with higher resting temperatures observed on days with higher daily maximum ambient temperatures but not to an extent that energy or water costs were significantly increased during rest. We found that, due to a relatively high level of thermal tolerance, high ambient temperatures are unlikely to cause an energetic strain on Southern Flying Squirrels. However, these findings do not preclude negative impacts of high ambient temperatures on the northern species, and these may still play a role in the changing distributions of Glaucomys in North America.
Las ardillas voladoras norteamericanas (Glaucomys sp.) han experimentado cambios dramáticos en su distribución hacia el norte durante las últimas décadas. Las investigaciones anteriores se han enfocado en los efectos potenciales del aumento de las temperaturas invernales, pero todavía no se ha explorado la hipótesis de que el aumento de la temperatura estival haya desempeñado un papel en estos cambios de rango. Por ende, deseamos determinar el efecto de las altas temperaturas ambientales sobre la termorregulación y la energía en las ardillas voladoras en una zona del noreste de América del Norte que ha mostrado un recambio de especies reciente. Estudiamos las ardillas voladoras del sur (Glaucomys volans), ya que no se pudo encontrar una población de la especie del norte (Glaucomys sabrinus) que fuera viable en términos de logística. Medimos la relación entre la tasa metabólica, la pérdida de agua por evaporación y la temperatura corporal a temperaturas ambiente altas por medio de la respirometría de flujo. Asímismo, determinamos la temperatura corporal interna de ardillas voladoras en libertad utilizando registradores de datos de temperatura. No detectamos ningún aumento estadísticamente significativo en la tasa metabólica hasta temperaturas ambiente de hasta 40 °C, pero la pérdida de agua por evaporación aumentó a temperaturas por encima de los 36,2 °C. La temperatura corporal de las ardillas voladoras en libertad siguió un patrón circadiano con una diferencia de ~2 °C entre las temperaturas corporales modales de la fase activa y de la fase de reposo. Las temperaturas ambientales afectaron las temperaturas de la fase de reposo. Los valores de dichas temperaturas fueron más altos en los días con temperaturas ambientales máximas diarias más elevadas, pero no se presentaron aumentos significativos en los costos de energía o agua durante el reposo. Determinamos que es poco probable que las altas temperaturas ambientales causen una tensión energética en las ardillas voladoras del sur debido a un nivel relativamente alto de tolerancia térmica. No obstante, estos hallazgos no excluyen los impactos negativos de las altas temperaturas ambientales sobre las especies del norte, que podrían tener un efecto sobre las distribuciones cambiantes de Glaucomys en América del Norte.

The intensification of agricultural practices and urbanisation are widespread causes of biodiversity loss. However, the role of artificial habitats in genetic rescue is an aspect that is not well understood. Implementing genetic rescue measures to improve gene flow and maintain a viable population of keystone species is a crucial prerequisite for promoting diverse and resilient ecosystems. Landscape fragmentation and modern agricultural methods have caused the decline and the isolation of the remnant colonies of the endangered European ground squirrel (Spermophilus citellus) throughout its range. However, the artificial habitat, such as airport fields with regular grass mowing, provides suitable conditions for this grassland specialist. We measured home range size and genetic variation of seven souslik colonies in western Slovakia. Based on the 6904 ddRAD SNPs, we found significantly higher individual heterozygosity in colonies on airports compared to colonies on pastures. This indicates a potential for higher fitness of individuals from airport colonies, which can serve as a source for evidence-based translocations. Such an intervention can preserve the genetic diversity of small and isolated populations in the region. We emphasize that conservation management strategies would be strengthened including a specific focus on human-made grassland habitats.

Mammalian hibernators survive prolonged periods of cold and resource scarcity by temporarily modulating normal physiological functions, but the mechanisms underlying these adaptations are poorly understood. The hibernation cycle of thirteen-lined ground squirrels (Ictidomys tridecemlineatus) lasts for 5-7 months and comprises weeks of hypometabolic, hypothermic torpor interspersed with 24-48-h periods of an active-like interbout arousal (IBA) state. We show that ground squirrels, who endure the entire hibernation season without food, have negligible hunger during IBAs. These squirrels exhibit reversible inhibition of the hypothalamic feeding center, such that hypothalamic arcuate nucleus neurons exhibit reduced sensitivity to the orexigenic and anorexigenic effects of ghrelin and leptin, respectively. However, hypothalamic infusion of thyroid hormone during an IBA is sufficient to rescue hibernation anorexia. Our results reveal that thyroid hormone deficiency underlies hibernation anorexia and demonstrate the functional flexibility of the hypothalamic feeding center.

The current study was conducted to determine the phylogroups and antibiotic susceptibilities of Escherichia coli isolates recovered from fecal samples of Anatolian Ground Squirrels (Spermophilus xanthoprymnus) and to examine the relationship between them. Eighty-two E. coli isolates obtained from 150 fecal samples were investigated. The quadruplex polymerase chain reaction (PCR), phylogroup C-, and E-specific mPCR were subjected to phylogenetic typing of the isolates. The susceptibilities to fifteen antibiotics of the isolates were detected by the disk diffusion method. In the result of phylogenetic typing, phylogroup B2 was most predominant (58.6 %), followed by B1 (25.6 %), E (8.5 %), C (4.9 %), and D (2.4 %). The phylogroup A, F, and Escherichia clades were not detected. The antibiotic susceptibility test revealed that 59.8 % (49/82) and 19.5 % (16/82) of E. coli isolates were resistant to at least one antibiotic and multidrug-resistant (MDR), respectively. Twenty-six (31.7 %), 19 (23.2 %), 11 (13.4 %), and 10 (12.2 %) of the isolates were found to be resistant to gentamicin, tetracycline, amoxicillin-clavulanic acid, and cefoxitin. Of the 49 E. coli isolates that were found to be resistant to any antibiotic analyzed, 30, 13, 4, and 2 were located in phylogroup B2, B1, E, and D, respectively. MDR isolates were mostly located in both phylogroup B1 (31.3 %) and B2 (31.3 %). In conclusion, data from the current study suggest that the isolates may potentially have pathogenic properties, since the majority (69.5 %) of E. coli isolates from fecal samples of Spermophilus xanthoprymnus were located in the pathogenic phylogroup and resistance to various antibiotics was detected.

Enterocytozoon bieneusi is an obligate intracellular microsporidian parasite with a worldwide distribution. As a zoonotic pathogen, E. bieneusi can infect a wide range of wildlife hosts through the fecal-oral route. Although the feces of flying squirrels (Trogopterus xanthipes) are considered a traditional Chinese medicine (as \"faeces trogopterori\"), no literature is available on E. bieneusi infection in flying squirrels to date. In this study, a total of 340 fresh flying squirrel fecal specimens from two captive populations were collected in Pingdingshan city, China, to detect the prevalence of E. bieneusi and assess their zoonotic potential. By nested PCR amplification of the ITS gene, six specimens tested positive, with positive samples from each farm, with an overall low infection rate of 1.8%. The ITS sequences revealed three genotypes, including known genotype D and two novel genotypes, HNFS01 and HNFS02. Genotype HNFS01 was the most prevalent (4/6, 66.7%). Phylogenetic analysis showed that all genotypes clustered into zoonotic Group 1, with the novel genotypes clustering into different subgroups. To our knowledge, this is the first report of E. bieneusi infection in flying squirrels, suggesting that flying squirrels could act as a potential reservoir and zoonotic threat for E. bieneusi transmission to humans in China.
UNASSIGNED: Occurrence et génotypage d’Enterocytozoon bieneusi chez les écureuils volants (Trogopterus xanthipes) de Chine.
UNASSIGNED: Enterocytozoon bieneusi est un parasite microsporidien intracellulaire obligatoire présent dans le monde entier. En tant qu’agent pathogène zoonotique, E. bieneusi peut infecter un large éventail d’hôtes sauvages par la voie fécale-orale. Bien que les excréments d’écureuils volants (Trogopterus xanthipes) soient considérés comme un ingrédient de médecine traditionnelle chinoise (comme « faeces trogopterori »), aucune littérature n’est disponible à ce jour sur l’infection par E. bieneusi chez les écureuils volants. Dans cette étude, un total de 340 spécimens fécaux frais d’écureuils volants provenant de deux populations captives ont été collectés dans la ville de Pingdingshan, en Chine, pour détecter la prévalence d’E. bieneusi et évaluer leur potentiel zoonotique. Par amplification PCR nichée du gène ITS, six échantillons se sont révélés positifs, avec des échantillons positifs dans chaque ferme, et un taux d’infection global faible, à 1,8 %. Les séquences ITS ont révélé trois génotypes, dont le génotype D connu et deux nouveaux génotypes, HNFS01 et HNFS02. Le génotype HNFS01 était le plus répandu (4/6, 66,7 %). L’analyse phylogénétique a montré que tous les génotypes se regroupaient dans le groupe zoonotique 1, les nouveaux génotypes se regroupant en différents sous-groupes. À notre connaissance, il s’agit du premier rapport d’infection par E. bieneusi chez des écureuils volants, ce qui suggère que les écureuils volants pourraient agir comme un réservoir potentiel et une menace zoonotique pour la transmission d’E. bieneusi aux humains en Chine.

Urban areas are expanding exponentially, leading more species of wildlife living in urban environments. Urban environmental characteristics, such as human disturbance, induce stress for many wildlife and have been shown to affect some cognitive traits, such as innovative problem-solving performance. However, because different cognitive traits have common cognitive processes, it is possible that urban environmental characteristics may directly and indirectly affect related cognitive traits (the ripple effect hypothesis). We tested the ripple effect hypothesis in urban Eurasian red squirrels residing in 11 urban areas that had different urban environmental characteristics (direct human disturbance, indirect human disturbance, areas of green coverage and squirrel population size). These squirrels were innovators who had previously repeatedly solved a food extraction task (the original task). Here, we examined whether and how urban environmental characteristics would directly and indirectly influence performance in two related cognitive traits, generalisation and (long-term) memory. The generalisation task required the innovators to apply the learned successful solutions when solving a similar but novel problem. The memory task required them to recall the learned solution of the original task after an extended period of time. Some of the selected urban environmental characteristics directly influenced the task performance, both at the population level (site) and at individual levels. Urban environmental characteristics, such as increased direct and indirect human disturbance, decreased the proportion of success in solving the generalisation task or the memory task at the population (site) level. Increased direct human disturbance and less green coverage increased the solving efficiency at individual levels. We also found an indirect effect in one of the urban environmental characteristics, indirect human disturbance, in the generalisation task, but not the memory task. Such an effect was only seen at the individual level but not at the population level; indirect human disturbance decreased the first original latency, which then decreased the generalisation latency across successes. Our results partially support the ripple effect hypothesis, suggesting that urban environmental characteristics are stressors for squirrels and have a greater impact on shaping cognitive performance than previously shown. Together, these results provide a better understanding of cognitive traits that support wildlife in adapting to urban environments.

BackgroundTo be better prepared for emerging wildlife-borne zoonoses, we need to strengthen wildlife disease surveillance.AimThe aim of this study was to create a topical overview of zoonotic pathogens in wildlife species to identify knowledge gaps and opportunities for improvement of wildlife disease surveillance.MethodsWe created a database, which is based on a systematic literature review in Embase focused on zoonotic pathogens in 10 common urban wildlife mammals in Europe, namely brown rats, house mice, wood mice, common voles, red squirrels, European rabbits, European hedgehogs, European moles, stone martens and red foxes. In total, we retrieved 6,305 unique articles of which 882 were included.ResultsIn total, 186 zoonotic pathogen species were described, including 90 bacteria, 42 helminths, 19 protozoa, 22 viruses and 15 fungi. Most of these pathogens were only studied in one single animal species. Even considering that some pathogens are relatively species-specific, many European countries have no (accessible) data on zoonotic pathogens in these relevant animal species. We used the Netherlands as an example to show how this database can be used by other countries to identify wildlife disease surveillance gaps on a national level. Only 4% of all potential host-pathogen combinations have been studied in the Netherlands.ConclusionsThis database comprises a comprehensive overview that can guide future research on wildlife-borne zoonotic diseases both on a European and national scale. Sharing and expanding this database provides a solid starting point for future European-wide collaborations to improve wildlife disease surveillance.

Thirteen-lined ground squirrels (TLGSs) are obligate hibernators that cycle between torpor (low metabolic rate and body temperature) and interbout euthermia (IBE; typical euthermic body temperature and metabolism) from late autumn to spring. Many physiological changes occur throughout hibernation, including a reduction in liver mitochondrial metabolism during torpor, which is reversed during arousal to interbout euthermia. Nuclear-encoded microRNA (miRNA, small posttranscriptional regulator molecules) differ in abundance throughout TLGS hibernation and have been shown to regulate mitochondrial gene expression in mammalian cell culture (where they are referred to as mitomiRs). This study characterized differences in mitomiR profiles from TLGS liver mitochondria isolated during summer, torpor, and IBE, and predicted their mitochondrial targets. Using small RNA sequencing, differentially abundant mitomiRs were identified between hibernation states, and using quantitative PCR analysis, we quantified the expression of predicted mitochondrial mRNA targets. Most differences in mitomiR abundances were seasonal (i.e., between summer and winter) with only one mitomiR differentially abundant between IBE and torpor. Multiple factor analysis (MFA) revealed three clusters divided by hibernation states, where clustering was predominantly driven by mitomiR abundances. Nine of these differentially abundant mitomiRs had predicted mitochondrial RNA targets, including subunits of electron transfer system complexes I and IV, 12S rRNA, and two tRNAs. Overall, mitomiRs were predicted to suppress the expression of their mitochondrial targets and may have some involvement in regulating protein translation in mitochondria. This study found differences in mitomiR abundances between seasons and hibernation states of TLGS and suggests potential mechanisms for regulating the mitochondrial electron transfer system.NEW & NOTEWORTHY During the hibernation season, thirteen-lined ground squirrels periodically increase metabolism remarkably between torpor and interbout euthermia (IBE). This process involves rapid reactivation of mitochondrial respiration. We predicted that mitochondrial microRNA (mitomiRs) might be altered during this response. We found that the abundance of 38 liver mitomiRs differs based on hibernation state (summer, IBE, and torpor). Small RNA sequencing identified mitomiR profiles, including some mitomiRs that are predicted to bind to mitochondrial RNAs.

Food hoarding provides animals access to resources during periods of scarcity. Studies on mammalian caching indicate associations with brain size, seasonality and diet but are biased to a subset of rodents. Whether the behaviour is generalizable at other taxonomic scales and/or is influenced by other ecological factors is less understood. Population density may influence food caching due to food competition or pilferage, but this remains untested in a comparative framework. Using phylogenetic analyses, we assessed the role of morphology (body and brain size), climate, diet breadth and population density on food caching behaviour evolution at multiple taxonomic scales. We also used a long-term dataset on caching behaviour of red squirrels (Tamiasciurus fremonti) to test key factors (climate and population density) on hoarding intensity. Consistent with previous smaller scale studies, we found the mammalian ancestral state for food caching was larderhoarding, and scatterhoarding was derived. Caching strategy was strongly associated with brain size, population density and climate. Mammals with larger brains and hippocampal volumes were more likely to scatterhoard, and species living at higher population densities and in colder climates were more likely to larderhoard. Finer-scale analyses within families, sub-families and tribes indicated that the behaviour is evolutionary labile. Brain size in family Sciuridae and tribe Marmotini was larger in scatterhoarders, but not in other tribes. Scatterhoarding in tribe Marmotini was more likely in species with lower population densities while scatterhoarding in tribe Sciurini was associated with warmer climates. Red squirrel larderhoarding intensity was positively related to population density but not climate, implicating food competition or pilferage as an important mechanism mediating caching behaviour. Our results are consistent with previous smaller-scale studies on food caching and indicate the evolutionary patterns of mammalian food caching are broadly generalizable. Given the lability of caching behaviour as evidenced by the variability of our results at finer phylogenetic scales, comparative analyses must consider taxonomic scale. Applying our results to conservation could prove useful as changes in population density or climate may select for different food caching strategies and thus can inform management of threatened and endangered species and their habitats.

This study investigates the adaptive strategies of the Alashan Ground Squirrel (Spermophilus alashanicus) in response to habitat changes, as rodents are sensitive indicators of ecosystem changes. Despite its ecological importance, the genome and microbiome of this species have not been thoroughly studied. This research fills this gap by presenting the first comprehensive metagenomic and transcriptomic datasets of the species. Transcriptomic data was collected from five tissue types, including heart, liver, cecum, muscle, and blood, resulting in the assembly of 72,156 unigenes. Metagenomic sequencing identified predominant bacterial groups such as Firmicutes, Bacteroidetes, Verrucomicrobia, Urovircota, and Proteobacteria. Our workflow involved RNA and DNA extraction, library preparation, assembly, and annotation, yielding valuable insights into gene discovery, microbial composition, and further genome and microbial function studies. In conclusion, our findings have significant implications for understanding the adaptive mechanisms of this species in response to environmental changes.