Comparative analysis of the gut microbiome of ungulate species from Qinghai-Xizang plateau.

Several studies have investigated the gut bacterial composition of wild ungulates in the Qinghai-Xizang Plateau. However, the relationship between their gut microbiome dendrograms and their phylogenetic tree remains unclear. In this study, we analyzed 45 amplicons (V3-V4 region of the 16S rRNA gene) from five wild ungulates-Pseudois nayaur, Pantholops hodgsonii, Gazella subgutturosa, Bos grunniens, and Equus kiang-from the Qinghai-Xizang Plateau to clarify the relationship between their phylogenies and gut microbiome dendrograms. The unweighted pair group method with arithmetic mean analysis and hierarchical clustering analysis indicated that G. subgutturosa is closely related to P. nayaur; however, these results were inconsistent with their phylogenetic trees. Additionally, the indicator genera in the microbiome of each wild ungulate showed strong associations with the diets and habitats of their host. Thus, diet and space niche differentiation may primarily account for the differences between the gut microbiome characteristics of these wild ungulates and their phylogeny. In summary, our research provides insights into the evolutionary factors influencing the gut microbiome of wild ungulates in the Qinghai-Xizang Plateau.

High toxinogenic potential of Staphylococcus aureus from wild ungulates in Brandenburg, Germany with a low level of antibiotic resistance.

Introduction: Data regarding the occurrence and virulence of Staphylococcus (S.) aureus in wild living animals is rare. However, S. aureus may carry a multitude of virulence factors and express resistance to several antimicrobial substances. Handling game meat may thus lead to serious infections or food poisoning. The aim of this study was to provide insights into the occurrence and characteristics of S. aureus in wild ungulates from Brandenburg, Germany. Methods: Nasal swabs of externally healthy-looking wild boars, roe, fallow and red deer were collected in hunts during season 2021/2022 and analyzed for S. aureus by selective enrichment. Species were determined using matrix assisted laser desorption ionization mass spectrometry and tested for phenotypic antimicrobial resistance. Whole-genome sequencing was conducted for genotyping, determination of virulence associated genes and analysis of phylogenetic relationships. Results: S. aureus were recovered from approximately 8% of nasal swabs. However, the strains were only obtained from the sampled wild ruminants. S. aureus isolates were associated with sequence types (ST) 1, ST30, ST133, ST425, ST582 and ST6238. Isolates of ST1 clustered closely together in the phylogenetic analysis. Genes encoding staphylococcal enterotoxin (SE) or SE-like (SEl) were found in 14/17 isolates. In particular, a seh gene was present in 12/17 isolates. Moreover, two isolates harbored a multiplicity of genes encoding SE or SEl. In addition, the toxic shock syndrome toxin encoding tst gene was detected in one isolate. This isolate was resistant to penicillin and cefoxitin and accordingly harbored the blaZ gene. Discussion: Wild ungulates intended for human consumption may carry potentially virulent S. aureus. In one case, the close phylogenetic relationship of S. aureus isolates indicates a possible intraspecific spread within a common territory. However, for others, the origin or the spread pattern can only be inferred. Handling of animals or their carcasses might contribute to staphylococcal infections in humans. Moreover, food poisoning due to SE producing strains may occur, if recommended hygiene practices are not applied during processing of game meat.

Epidemiological factors conditioning sarcoptic mange spreading in wild boar (Sus scrofa).

Sarcoptic mange is a widely distributed disease, with numerous potential hosts among domestic and wild animals. Nowadays it is considered a neglected re-emergent infection in humans. As a difference with domestic pigs, and even with several clinical cases reported in some European countries, it seems that Eurasian wild boars (Sus scrofa) have a low susceptibility to clinical mange. However, because of a case of confirmed transmission from Spanish ibex (Capra pyrenaica) to wild boar in the province of Tarragona, we planned a large-scale ELISA survey in the neighboring Valencian Community (SE Spain). We compared 419 wild boar sera from different management systems (fenced vs. open game estates), different ages (piglets, juveniles, and adults), with different behaviour (gregarious females of all ages and male piglets vs. solitary juveniles and adult males), from areas with different wild boar densities, different wild ruminant densities and different sarcoptic mange epidemiologic situations. The whole prevalence of antibodies against sarcoptic mange in the tested wild boars was 10.5%. No significant differences were found when comparing fenced and free ranging wild boars, males and females, gregarious vs. solitary individuals or among different ages. However, wild boar density was a relevant factor. In areas with a hunting bag of <1 wild boar/km2, considered as a low density of suids, the seroprevalence was 2.94%, but rose to 11.52% in high density districts, constituting a significant difference (p = 0.037). Low wild boar populations would act as a protective factor (OR 0.233; p = 0.049) against coming into contact with the mite. The wild ruminant densities or their sarcoptic mange status did not show any effect on wild boars seroprevalence against this disease. These results reinforce the suggested host-taxon Sarcoptes scabiei specificity and the independence of host-species foci.

Fecal thyroid hormone metabolites in wild ungulates: a mini-review.

This review aims to analyse the fluctuations of fecal thyroid hormone metabolites (FTMs) related to environmental and individual variables in different species of wild ungulates and provide a collection of assay methods. The great advantage of fecal sampling is being completely non-invasive. A systemic search was conducted from 2019 to 2024, using data sources PubMed, Scopus, Web of Science, and the World Wide Web, and ten studies were found on this topic. Three studies used the radioimmunoassay method for FTMs analysis, while the others used a less expensive enzyme-linked immunosorbent assay. Most of these papers validated the method for the species-specific matrix. Related to the studied variables, some authors analysed FTM fluctuations only concerning individual variables, and others in response to both. Temperature and fecal cortisol metabolites (FCMs) were the most studied environmental and individual variables, respectively. Since FTMs are an integrative measure of plasma thyroid hormones, the information obtained from a non-invasive-assay method regarding wild ungulate physiology is becoming of great interest to the scientific community.

Seeds in the guts: can seed traits explain seed survival after being digested by wild ungulates?

Plants inhabiting open landscapes are often dispersed by ungulates and are expected to be adapted to this type of dispersal through their seed traits. To find which traits help seeds survive the passage through digestion of wild ungulates, we conducted a comprehensive feeding experiment with almost forty species of plants and three species of ungulates. We fed specified numbers of seeds to the animals, collected the dung, and germinated the dung content. We explored whether seed morphological traits and seed nutrient contents are good predictors of seed survival after passage through the ungulate digestive system. We also tested how the seed survival differed after the passage through different ungulate species. To find answers, we used GLMM with beta-binomial distribution and animal and plant species as random factor, respectively. We found that species survival and germination success were negatively correlated to seed elongation and the thickness of the seed coat. Even though phylogenetically correct GLMM did not yield significant results, when we tested species from commonly represented families, separately (legumes and grasses compared to all other species) different traits had statistically significant effects. In the case of seed elongation, the effect changed direction from negative to positive when legumes and grasses were left out. Our results suggest that seed traits enabling species survival after passage through the digestive tract are strongly phylogenetically conserved and different groups of plants evolved different ways of adapting to grazing pressure and utilize it for dispersal.

Gastrointestinal parasites of the wild ungulates (Mammalia: Cetartiodactyla) in the Hemis National Park, Ladakh, India.

We evaluated the gastrointestinal parasitic prevalence and intensity of wild ungulates in the Hemis National Park of Ladakh by examining their fresh faecal samples. A total of 327 fresh faecal samples from three wild ungulates, blue sheep Pseudois nayaur (n = 127), Ladakh urial Ovis vignei vignei (n = 110), and Himalayan ibex Ibex siberica (n = 90) were collected between June 2021 and May 2022. The techniques of flotation and sedimentation were used to find parasite eggs and oocysts from the faecal samples. Out of 327 samples examined, 165 samples were infected with gastrointestinal parasites making an overall prevalence of 50.45%. Seven parasitic taxa, including one protozoan (Eimeria spp.), five nematodes (Nematodirus spp., Strongyloides spp., Haemonchus sp., Trichuris sp., and Trichostrongylus spp.), and one cestode (Monezia spp.), were found during the current investigation. Mixed infection was reported in 36 (11%) of the total examined samples. The prevalence of gastrointestinal parasites was found to be highest in blue sheep (55.11%), followed by Ladakh urial (49.09%) and Himalayan ibex (45.55%). The highest prevalence was recorded in the summer (64.42%), and the lowest in the winter (33.82%). A significant difference in parasitic prevalence was observed across seasons in each of the three wild ungulate hosts. However, there was no significant difference in the prevalence of parasites between these hosts.

Circulation of picobirnavirus in Neotropical free-ranging mammals.

Picobirnavirus (PBV) is a family of non-enveloped double-stranded RNA viruses with bisegmented genomes. Segment 1 encodes the capsid protein and segment 2 encodes RNA-dependent RNA polymerase. They exhibit high genomic heterogeneity and infect a wide range of vertebrate hosts, including humans. The objective of this study was to expand our knowledge of the circulation of PBV in free-living animals from two regions (Brazil and Argentina) of the Atlantic Forest. Fecal samples were analyzed from free-living animals: tapir, brocket deer, peccary, and different species of rodents and marsupials. A total of 133 samples were collected and analyzed by RT-PCR, of which 44 (33.08%) were PBV-positive. Nine amplicons were sequenced, five species from Argentina and four from Brazil, and phylogenetic analysis was performed. The nucleotide and amino acid identities of the PBV strains detected in animals from Argentina and Brazil were between 66.3% and 82.5% and between 55.3% and 74.2%, respectively. The analysed strains presented conserved nucleotide blocks without distinction of the host species. The phylogenetic tree showed that PBV strains from Atlantic Forest animals belonging to genogroup I were grouped into different clusters, without defining groups according to host species (human or animal) or the geographical area of detection. This is the first study on PBV in free-living animals in the Atlantic Forest. Our analysis suggested that PBV strains can infect different animal species, leading to PBV transmission between animals and humans. This reinforces the hypothesis of previous crossover points in the ecology and evolution of heterologous PBV strains.

Wild Ungulates Constitute the Basis of the Diet of the Iberian Wolf in a Recently Recolonized Area: Wild Boar and Roe Deer as Key Species for Its Conservation.

The Iberian wolf (Canis lupus signatus) is recolonizing historical distribution areas after decades of absence. As in other human-dominated landscapes, finding a balance to protect this species by favoring recolonization and mitigating human-wildlife conflicts is a challenge. Since wolves are often generalist opportunistic predators, we studied their diet composition in central Spain to evaluate the consumption of domestic ungulates and provide reliable data that could help local authorities to deal with the current wolf-cattle ranchers conflict and coexistence. Diet composition (% prey occurrence, % prey ingested biomass) was analyzed through the identification of prey hairs present in 671 scats collected between 2017 and 2021. The wolves fed more on wild ungulates (82% occurrence) than domestic ones (18%). Wild boar (Sus scrofa, 44% occurrence) and roe deer (Capreolus capreolus, 35%) were the most consumed prey. The wolves positively selected these two species. The wolves' diets varied between seasons, years, and forest regions, but a diet based on wild ungulates predominated over domestic ones. Food niche breadth showed variations depending on seasons and years. Preserving the availability and diversity of wild ungulates may favor reducing livestock attacks and would be an achievable goal that would help to conserve this species and reduce conservation conflicts.

Parasite-mediated manipulation? Toxoplasma gondii infection increases risk behaviour towards culling in red deer.

Parasites can modify host behaviour to increase their chances of survival and transmission. Toxoplasma gondii is a globally distributed protozoan whose ability to modify host behaviour is well known in taxa such as rats and humans. Less well known are the effects on the behaviour of wild species, with the exception of a few studies on primates and carnivores. Taking advantage of a culling activity conducted in Stelvio National Park (Italy), the serological status of T. gondii was studied in 260 individuals of red deer Cervus elaphus with respect to the risk of being culled. A temporal culling rank index was fitted as a response variable, and T. gondii serological status as the main explanatory variable in linear models, accounting for covariates such as sex, age, jaw length, bone marrow fat and culling location. The overall seroprevalence of T. gondii was 31.5%, and the selected models suggested that seropositive deer were culled earlier than seronegative ones, but this effect was only evident in females, in individuals with medium-good body condition, and in areas with greater human presence. Our results suggest that T. gondii may be involved in risk behaviour in large herbivores, supporting its role as a facilitator of predation risk.

Wild ungulates as sentinels of flaviviruses and tick-borne zoonotic pathogen circulation: an Italian perspective.

BACKGROUND: Vector-borne zoonotic diseases are a concerning issue in Europe. Lyme disease and tick-borne encephalitis virus (TBEV) have been reported in several countries with a large impact on public health; other emerging pathogens, such as Rickettsiales, and mosquito-borne flaviviruses have been increasingly reported. All these pathogens are linked to wild ungulates playing roles as tick feeders, spreaders, and sentinels for pathogen circulation. This study evaluated the prevalence of TBEV, Borrelia burgdorferi sensu lato, Rickettsia spp., Ehrlichia spp., and Coxiella spp. by biomolecular screening of blood samples and ticks collected from wild ungulates. Ungulates were also screened by ELISA and virus neutralization tests for flaviviral antibody detection. RESULTS: A total of 274 blood samples were collected from several wild ungulate species, as well as 406 Ixodes ricinus, which were feeding on them. Blood samples tested positive for B. burgdorferi s.l. (1.1%; 0-2.3%) and Rickettsia spp. (1.1%; 0-2.3%) and showed an overall flaviviral seroprevalence of 30.6% (22.1-39.2%): 26.1% (17.9-34.3%) for TBEV, 3.6% (0.1-7.1%) for Usutu virus and 0.9% (0-2.7%) for West Nile virus. Ticks were pooled when possible and yielded 331 tick samples that tested positive for B. burgdorferi s.l. (8.8%; 5.8-11.8%), Rickettsia spp. (26.6%; 21.8-31.2%) and Neoehrlichia mikurensis (1.2%; 0-2.4%). TBEV and Coxiella spp. were not detected in either blood or tick samples. CONCLUSIONS: This research highlighted a high prevalence of several tick-borne zoonotic pathogens and high seroprevalence for flaviviruses in both hilly and alpine areas. For the first time, an alpine chamois tested positive for anti-TBEV antibodies. Ungulate species are of particular interest due to their sentinel role in flavivirus circulation and their indirect role in tick-borne diseases and maintenance as Ixodes feeders and spreaders.

The method matters. A comparative study of biologging and camera traps as data sources with which to describe wildlife habitat selection.

Habitat use is a virtually universal activity among animals and is highly relevant as regards designing wildlife management and conservation actions. This has led to the development of a great variety of methods to study it, of which resource selection functions combined with biologging-derived data (RSF) is the most widely used for this purpose. However this approach has some constraints, such as its invasiveness and high costs. Analytical approaches taking into consideration imperfect detection coupled with camera trap data (IDM) have, therefore, emerged as a non-invasive cost-effective alternative. However, despite the fact that both approaches (RSF and IDM) have been used in habitat selection studies, they should also be comparatively assessed. The objective of this work is consequently to assess them from two perspectives: explanatory and predictive. This has been done by analyzing data obtained from camera traps (60 sampling sites) and biologging (17 animals monitored: 7 red deer Cervus elaphus, 6 fallow deer Dama dama and 4 wild boar Sus scrofa) in the same periods using IDM and RSF, respectively, in Doñana National Park (southern Spain) in order to explain and predict habitat use patterns for three studied species. Our results showed discrepancies between the two approaches, as they identified different predictors as being the most relevant to determine species intensity of use, and they predicted spatial patterns of habitat use with a contrasted level of concordance, depending on species and scale. Given these results and the characteristics of each approach, we suggested that although partly comparable interpretations can be obtained with both approaches, they are not equivalent but rather complementary. The combination of data from biologging and camera traps would, therefore, appear to be suitable for the development of an analytical framework with which to describe and characterise the habitat use processes of wildlife.

[Circulation of bovine herpesvirus (Herpesviridae: Varicellovirus) and bovine viral diarrhea virus (Flaviviridae: Pestivirus) among wild artiodactyls of the Moscow region].

INTRODUCTION: Pestiviruses and viruses of the Herpesviridae family are widely distributed among different species of ungulates, but the main information about these pathogens is related to their effect on farm animals. Data on detection of bovine viral diarrhea virus (BVDV) and bovine herpes virus (BoHV) in wild ungulates reported from different countries in recent years raises the question of the role of wild animals in the epidemiology of cattle diseases. AIM OF WORK: To study the prevalence of herpesviruses and pestiviruses in the population of wild artiodactyls of the Moscow region. MATERIALS AND METHODS: Samples of parenchymal organs and mucosal swabs from 124 wild deer (moose and roe deer) shot during hunting seasons 20192022 in Moscow Region were examined by PCR, virological and serological methods for the presence of genetic material and antibodies to bovine infectious rhinotracheitis and viral diarrhea. RESULTS: BVDV RNA was found in a sample from one moose, BoHV DNA was detected in samples from three roe deer and two moose shot in the Moscow region. Seropositive animals were of different sex and age, the total BoHVs and BVDV seroprevalence rates in wild artiodactyls were 46 and 29%, respectively. CONCLUSION: Wild ruminant artiodactyls of the Moscow Region can be a natural reservoir of BoHV-1, and this must be taken into account when planning and organizing measures to control the infectious bovine rhinotracheitis. Cases of BVDV infection in wild artiodactyls are less common, so more research is needed to definitively establish their role in the epidemiology of this disease in cattle.

Different Methods for Seminal Plasma Removal and Sperm Selection on the Quality and Fertility of Collared Peccary Sperm.

Methods for seminal plasma (SP) removal and the selection of collared peccary sperm for fertilization were compared. The experiments evaluated the following: the (I) impact of centrifugation for SP removal before swim-up for sperm selection and (II) a comparison of different Percoll® gradient densities (PG 45-90% and PG 35-70%). Non-selected sperm served as the control. Sperm quality was assessed based on motility patterns, morphology, membrane functional integrity, viability, reactive oxygen species (ROS), glutathione (GSH), and DNA integrity. Subsequently, the most successful group in the previous experiment and washing by centrifugation (WC) were compared for motility patterns and fertilization using pig oocytes. Swim-up decreased motility and enhanced ROS compared to the control. Centrifugation before swim-up harmed integrity and viability compared to the control. PG 45-90% (96.8 vs. 69.7 vs. 40.7 µm/s) allowed for a better velocity average pathway (VAP), a better velocity straight line, and better linearity (LIN) than those of the control and PG 35-70% (88.4 vs. 56.0 vs. 27.3 µm/s). Thus, PG 45-90% was used for fertilization. PG 45-90% obtained a higher VAP, a higher amplitude of the lateral head, straightness, and higher LIN than those of the control and WC. Cleavage (25.2-26.3%) and morula (8.1-10.5%) rates did not differ between the groups. Therefore, PG 45-90% and WC were efficient in isolating collared peccary sperm capable of fertilizing pig oocytes.

Epidemiological Dynamics of Foot-and-Mouth Disease in the Horn of Africa: The Role of Virus Diversity and Animal Movement.

The Horn of Africa is a large area of arid and semi-arid land, holding about 10% of the global and 40% of the entire African livestock population. The region's livestock production system is mainly extensive and pastoralist. It faces countless problems, such as a shortage of pastures and watering points, poor access to veterinary services, and multiple endemic diseases like foot-and-mouth disease (FMD). Foot-and-mouth disease is one of the most economically important livestock diseases worldwide and is endemic in most developing countries. Within Africa, five of the seven serotypes of the FMD virus (FMDV) are described, but serotype C is not circulating anymore, a burden unseen anywhere in the world. The enormous genetic diversity of FMDV is favored by an error-prone RNA-dependent RNA polymerase, intra-typic and inter-typic recombination, as well as the quasi-species nature of the virus. This paper describes the epidemiological dynamics of foot-and-mouth disease in the Horn of Africa with regard to the serotypes and topotypes distribution of FMDV, the livestock production systems practiced, animal movement, the role of wildlife, and the epidemiological complexity of FMD. Within this review, outbreak investigation data and serological studies confirm the endemicity of the disease in the Horn of Africa. Multiple topotypes of FMDV are described in the literature as circulating in the region, with further evolution of virus diversity predicted. A large susceptible livestock population and the presence of wild ungulates are described as complicating the epidemiology of the disease. Further, the husbandry practices and legal and illegal trading of livestock and their products, coupled with poor biosecurity practices, are also reported to impact the spread of FMDV within and between countries in the region. The porosity of borders for pastoralist herders fuels the unregulated transboundary livestock trade. There are no systematic control strategies in the region except for sporadic vaccination with locally produced vaccines, while literature indicates that effective control measures should also consider virus diversity, livestock movements/biosecurity, transboundary trade, and the reduction of contact with wild, susceptible ungulates.

Basic Hazard Control Plan for Small Wild Ungulates Slaughtered for Meat Production.

Animal slaughter plans and related activities must not increase the number of hazards in meat. In their nature, these processes must reduce possible hazards to minimum or acceptable levels. This is a generally accepted concept worldwide; hence, authorities continue to develop regulations that seek to mitigate the scourge of meat hazards for consumer protection. The situation is similar with small wild ungulate meat, in which a hazard analysis plan needs developing to improve meat safety. This investigation follows a narrative review of articles published for a PhD program and other scholarly articles supporting the concept of a basic slaughter plan for small wild ungulate animals in South Africa. The findings of this investigation highlight the need to control hazards within one health concept plan, which should be implemented and propagated by establishing forums that will drive meat safety solutions in these communities. There should be a basic hygiene slaughter plan developed and endorsed by all members of the forum. The outcome must be the control of microbiological, chemical and physical hazards from farm-to-fork, and as part of a system imbedded in game meat policies and regulations.

Seasonal Variation in the Body Composition, Carcass Composition, and Offal Quality in the Wild Fallow Deer (Dama dama L.).

The goal of this study was to examine the seasonal variation in the body composition, carcass composition, and quality of edible internal organs from the fallow deer hunt-harvested in the summer (n = 9) and the winter (n = 10) seasons. The weight and proportion of the mesenteric and omental fat were greater for the animals harvested in the winter (1.12 kg and 2.75%) compared to those from the summer season (0.43 kg and 1.02%). The winter-harvested animals had more perinephric fat (0.75 kg and 1.84%) than those hunted in summer (0.26 kg and 1.84%). The gastrointestinal tract of the fallow deer hunted in summer was more filled with feed and therefore heavier (7.92 kg) compared to those from the winter season (5.16 kg). The proportion of fat was significantly greater in the carcasses obtained in winter compared to the summer season (6.55% vs. 3.79%). No seasonal variety was found in the physicochemical characteristics of the edible offal, but the content of extractable fat was significantly affected by the season. In conclusion, the effect of the season on the slaughter value of the hunt-harvested fallow deer was limited to a variation in the proportion of some of the internal organs and affected the fat deposition in the body of the examined animals. The season significantly affected the fat content in the carcass and the extractable fat content in the examined offal.

Feeding Ecology of the Cuvier's Gazelle (Gazella cuvieri, Ogilby, 1841) in the Sahara Desert.

Knowledge of the feeding ecology of ungulates in arid biomes offers an interesting model for understanding the drought resistance of large desert-adapted herbivores, a crucial issue in the face of increasing desertification due to climate change. To assess the feeding ecology of the endangered Cuvier's gazelle (Gazella cuvieri) in the Sahara desert, we used a multi-method approach combining faecal samples, direct observations, and the recording of indirect signs of feeding. We hypothesised that browser behaviour is the best foraging strategy for species living in hyper-arid environments, mainly due to long periods without grazing opportunities. Complementarily, we explored the effects of the main environmental descriptors (rainfalls and NDVI) on feeding patterns and diet quality. We found that Cuvier's diets are based mainly on acacias (Vachellia tortilis, V. flava) and occasionally on the annual forb Anastatica hierochuntica. In total, eighteen species (five trees, nine shrubs, three herbs, and one grass) belonging to fifteen families were recorded. Our result confirmed the browsers' characteristic of this species, reaffirming its ability to settle in a hostile environment. Acacias stand out as key species consumed at the southernmost limit of their range; hence, future conservation plans and strategies should take this into account for the survival of Cuvier's gazelle in desert environments.

Occurrence and molecular characterization of Enterocytozoon bieneusi in wild and domestic animal species in Portugal.

The phylum Microsporidia encompasses a diverse group of obligate, intracellular, and spore-forming organisms able to infect a wide range of animal hosts. Among them, Enterocytozoon bieneusi is the most frequently reported species in humans and animals. Little is known about the presence and epidemiology of E. bieneusi in wildlife. We investigated E. bieneusi occurrence and genetic diversity in wild and domestic mammals, through molecular-detection methods, from different regions across Portugal. A total of 756 samples were collected from 288, 242, and 226 wild carnivores, wild ungulates, and domestic animals, respectively. Overall, eight specimens were E. bieneusi-positive (1.1%, 8/756) obtained from five wild (Iberian lynx, Iberian wolf, red fox, stone marten, and wild boar) and one domestic (sheep) host. Nucleotide sequence analysis identified four genotypes of E. bieneusi, Type IV, Wildboar3, BEB6, and PtEbIX. Three of those genotypes belong to Groups 1 (Type IV and Wildboar3) and 2 (BEB6), which are known to contain genotypes capable of infecting a variety of hosts, including humans, highlighting their public health importance. PtEbIX belongs to the dog-specific Group 11. This study represents the first, largest, and most comprehensive molecular-based epidemiology survey carried out in Portugal in wild and domestic animals to date and the first worldwide identification of E. bieneusi in wolf species. Our study showed that wild carnivores and ungulates may act as reservoirs of zoonotic genotypes of E. bieneusi, establishing their role in maintaining the sylvatic cycle of this parasite while representing a potential source of infection for humans and domestic animals.

The identification of human-pathogenic genotypes of fungi-related Enterocytozoon bieneusi in wild carnivores and ungulates in Portugal suggests cross-species infection events and overlapping of the sylvatic and domestic transmission cycles, demonstrating a potential transmission risk to humans.

The Impact of Variation in the Toll-like Receptor 3 Gene on Epizootic Hemorrhagic Disease in Illinois Wild White-Tailed Deer (Odocoileus virginianus).

Epizootic hemorrhagic disease (EHD) leads to high mortality in white-tailed deer (Odocoileus virginianus) and is caused by a double-stranded RNA (dsRNA) virus. Toll-like receptor 3 (TLR3) plays a role in host immune detection and response to dsRNA viruses. We, therefore, examined the role of genetic variation within the TLR3 gene in EHD among 84 Illinois wild white-tailed deer (26 EHD-positive deer and 58 EHD-negative controls). The entire coding region of the TLR3 gene was sequenced: 2715 base pairs encoding 904 amino acids. We identified 85 haplotypes with 77 single nucleotide polymorphisms (SNPs), of which 45 were synonymous mutations and 32 were non-synonymous. Two non-synonymous SNPs differed significantly in frequency between EHD-positive and EHD-negative deer. In the EHD-positive deer, phenylalanine was relatively less likely to be encoded at codon positions 59 and 116, whereas leucine and serine (respectively) were detected less frequently in EHD-negative deer. Both amino acid substitutions were predicted to impact protein structure or function. Understanding associations between TLR3 polymorphisms and EHD provides insights into the role of host genetics in outbreaks of EHD in deer, which may allow wildlife agencies to better understand the severity of outbreaks.

Phylodynamic Analysis Suggests That Deer Species May Be a True Reservoir for Hepatitis E Virus Genotypes 3 and 4.

Hepatitis E virus (HEV) genotypes 3 and 4 (HEV-3 and HEV-4) cause zoonotic infection in humans, with domestic pigs and wild boars being the main reservoirs of infection. Other than suids, HEV-3 and HEV-4 are found in ruminants, most frequently in deer species. However, it is still debatable, whether HEV infection in deer is a spillover, or indicates a stable virus circulation in these host species. To explore the patterns of HEV-3 and HEV-4 transmission in deer and other host species, we performed a Bayesian analysis of HEV sequences available in GenBank. A total of 27 HEV sequences from different deer species were found in GenBank. Sequences from wild boars collected in the same territories, as well as sequences from all mammals that were most similar to sequences from deer in blast search, were added to the dataset, comprising 617 in total sequences. Due to the presence of partial genomic sequences, they were divided into four subsets (two ORF1 fragments and two ORF2 fragments) and analyzed separately. European HEV-3 sequences and Asian HEV-4 sequences collected from deer species demonstrated two transmission patterns. The first pattern was spillover infection, and the second pattern was deer-to-deer transmission, indicating stable HEV circulation in these species. However, all geographic HEV clusters that contained both deer and swine sequences originated from ancestral swine strains. HEV-3 and HEV-4 transmission patterns in ungulates reconstructed by means of Bayesian analysis indicate that deer species are a true host for HEV. However, wild and domestic swine are often the primary source of infection for ruminants living in the same areas. Complete HEV genomic sequences from different parts of the world are crucial for further understanding the HEV-3 and HEV-4 circulation patterns in wildlife.