Universal versus taxon-specific drivers of helminth prevalence and intensity of infection.

Two key epidemiological parameters, prevalence and mean intensity of infection, together capture the abundance of macroparasite populations, the strength of density-dependent effects they experience, their potential impact on host population dynamics and the selective pressures they exert on their hosts. Yet, the drivers of the extensive variation observed in prevalence and mean intensity of infection, even among related parasite taxa infecting related hosts, remain mostly unknown. We performed phylogenetically grounded Bayesian modelling across hundreds of amphibian populations to test the effects of various predictors of prevalence and intensity of infection by six families of helminth parasites. We focused on the potential effects of key host traits and environmental factors pertinent to focal host populations, i.e. the local diversity of the amphibian community and local climatic variables. Our analyses revealed several important determinants of prevalence or intensity of infection in various parasite families, but none applying to all families. Our study uncovered no universal driver of parasite infection levels, even among parasite taxa from the same phylum, or with similar life cycles and transmission modes. Although local variables not considered here may have effects extending across taxa, our findings suggest the need for a taxon-specific approach in any attempt to predict disease dynamics and impacts in the face of environmental and climatic changes.

Assessing the impact of preventative measures to limit the spread of Toxoplasma gondii in wild carnivores of Madagascar.

Novel multihost pathogens can threaten endangered wildlife species, as well as humans and domestic animals. The zoonotic protozoan parasite Toxoplasma gondii is transmitted by members of Felidae and can infect a large number of animal species, including humans. This parasite can have significant health consequences for infected intermediate hosts and could further endanger wild carnivore populations of Madagascar. Building on an empirical characterization of the prevalence of the pathogen in local mammals, we used mathematical models of pathogen transmission in a multihost community to compare preventative measures that aim to limit the spread of this parasite in wild carnivores. Specifically, we examined the effect of hypothetical cat vaccination and population control campaigns on reducing the risk of infection by T. gondii in wild Eupleridae. Our model predicted that the prevalence of exposure to T. gondii in cats would be around 72% and that seroprevalence would reach 2% and 43% in rodents and wild carnivores, respectively. Reducing the rodent population in the landscape by half may only decrease the prevalence of T. gondii in carnivores by 10%. Similarly, cat vaccination and reducing the population of definitive hosts had limited impact on the prevalence of T. gondii in wild carnivorans of Madagascar. A significant reduction in prevalence would require extremely high vaccination, low turnover, or both in the cat population. Other potential control methods of T. gondii in endangered Eupleridae include targeted vaccination of wild animals but would require further investigation. Eliminating the threat entirely will be difficult because of the ubiquity of cats and the persistence of the parasite in the environment.

Evaluación del impacto de las medidas preventivas para limitar el contagio de Toxoplasma gondii en los carnívoros silvestres de Madagascar Resumen Los patógenos novedosos con múltiples hospederos pueden amenazar tanto a las especies silvestres como a los humanos y a los animales domésticos. Los miembros de la familia Felidae transmiten el protozoario parásito Toxoplasma gondii, el cual puede infectar a un gran número de especies animales, incluyendo al humano. Este parásito puede generar consecuencias importantes para la salud en los hospederos intermediarios infectados y podría poner más en peligro a las poblaciones de carnívoros silvestres de Madagascar. Usamos modelos matemáticos de la transmisión de patógenos en una comunidad con múltiples hospederos a partir de una caracterización empírica de la prevalencia del patógeno en los mamíferos locales para comparar las medidas preventivas que buscan limitar la transmisión de este parásito en los carnívoros silvestres. En específico, examinamos el efecto de la vacunación hipotética de felinos y las campañas de control poblacional sobre la reducción del riesgo de infección de T. gondii en los Eupleridae silvestres. Nuestro modelo predijo que la prevalencia de la exposición a T. gondii en los felinos sería de un 72% y que la seroprevalencia llegaría al 2% y al 43% en los roedores y carnívoros silvestres, respectivamente. La reducción a la mitad de la población de roedores en el paisaje podría disminuir sólo en un 10% la prevalencia del protozoario en los carnívoros. De forma similar, la vacunación y la reducción de la población de hospederos definitivos tuvieron un impacto limitado sobre la prevalencia de T. gondii en los carnívoros silvestres de Madagascar. Una reducción significativa en la prevalencia requeriría que la población de felinos tuviera una vacunación extremadamente elevada, baja rotación, o ambas. Otros métodos potenciales de control de T. gondii en los Eupleridae incluyen la vacunación de animales silvestres, pero requieren de mayor investigación. La eliminación completa de la amenaza será difícil por la ubicuidad de los felinos y la persistencia del parásito en el ambiente.

Scavenger-induced scattering of wild boar carcasses over large distances and its implications for disease management.

Vertebrate scavengers provide essential ecosystem services such as accelerating carrion decomposition by consuming carcasses, exposing tissues to microbial and invertebrate decomposers, and recycling nutrients back into the ecosystem. Some scavengers do not consume carcasses on site but rather scatter their remains in the surroundings, which might have important implications for nutrient transport, forensic investigations and the spread of diseases such as African Swine Fever. However, only a few studies have investigated and measured the scatter distances. Using wild boar (Sus scrofa) carcasses and limbs, we monitored scavenging behavior and measured scatter distances of mammals. We placed 20 carcasses (up to 25 kg) and 21 separate limbs equipped with very high frequency (VHF) transmitters and monitored scavenger activity using camera traps in a mountainous region in southeast Germany. Except for one carcass, all other carcasses and limbs were scattered. We measured 72 scatter distances (of 89 scattering events; mean = 232 m, maximum = 1250 m), of which 75% were dispersed up to 407 m. Scavengers moved scattered pieces into denser vegetation compared to the half-open vegetation at provisioning sites. Red foxes (Vulpes vulpes) were the most common scavenger species, contributing to 72 scattering events (58 measured scatter distances). Our results provide evidence of scatter distances farther than previously assumed and have far-reaching implications for disease management or forensic investigations, as the broader surroundings of carcasses must be included in search efforts to remove infectious material or relevant body parts for forensic analysis.

Leucism: the prevalent congenital malformation in the olive ridley sea turtle of northwestern Mexico.

Despite being the most abundant sea turtle in the world, the olive ridley turtle Lepidochelys olivacea is classified as Vulnerable by the IUCN. There is evidence of congenital malformations in hatchlings, and the associated causes are multifactorial, with both genetic and environmental sources. Santuario Playa Ceuta (SPC) is a sanctuary for the olive ridley, located at the northernmost region of its nesting range in the Mexican Pacific. The objective of this study was to identify and quantify the prevalence and severity of congenital malformations in olive ridley embryos/hatchlings in SPC during the 2017 nesting season. We collected 62907 eggs from 643 relocated nests that were moved to a hatchery, of which 4242 eggs with obvious development did not hatch and were analyzed for this study. Hatching success was 53.9%, with 22.5% of nests (n = 145) and 0.54% of eggs (n = 344) showing embryos or hatchlings with malformations. The nest severity index was 2.4 (range: 1-10) malformed embryos or hatchlings per nest, and the organism severity index was 1.4 (range: 1-7) malformations per malformed embryo or hatchling. Leucism was the most prevalent malformation (34.4%; 170/494 total observed), with the craniofacial region showing the greatest diversity of malformations (17/35 types). Given the geographical position of SPC, extreme environmental conditions (e.g. cold, heat, and dryness) could be one of the main causes of teratogenesis in this species. However, more studies are needed regarding the presence of contaminants, genetic factors, health assessments of nesting females, and malformation rates of nests that remain in situ versus those that are relocated.

Forest cover influences chytrid infections in populations of Boana curupi, a threatened treefrog of south Brazil.

Complex interactions among hosts, pathogens, and the environment affect the vulnerability of amphibians to the emergence of infectious diseases such as chytridiomycosis, caused by Batrachochytrium dendrobatidis (Bd). Boana curupi is a forest-dwelling amphibian endemic to the southern Atlantic Forest of South America, a severely fragmented region. Here, we evaluated whether abiotic factors (including air and water temperature, relative air humidity, and landscape) are correlated with chytrid infection intensity and prevalence in B. curupi. We found individuals infected with Bd in all populations sampled. Prevalence ranged from 25-86%, and the infection burden ranged from 1 to over 130000 zoospore genomic equivalents (g.e.) (mean ± SD: 4913 ± 18081 g.e.). The infection load differed among populations and was influenced by forest cover at scales of 100, 500, and 1000 m, with the highest infection rates recorded in areas with a higher proportion of forest cover. Our results suggest that the fungus is widely distributed in the populations of B. curupi in southern Brazil. Population and disease monitoring are necessary to better understand the relationships between host, pathogen, and environment, especially when, as in the case of B. curupi, threatened species are involved.

An agent-based framework for improving wildlife disease surveillance: A case study of chronic wasting disease in Missouri white-tailed deer.

Epidemiological surveillance for important wildlife diseases often relies on samples obtained from hunter-harvested animals. A problem, however, is that although convenient and cost-effective, hunter-harvest samples are not representative of the population due to heterogeneities in disease distribution and biased sampling. We developed an agent-based modeling framework that i) simulates a deer population in a user-generated landscape, and ii) uses a snapshot of the in silico deer population to simulate disease prevalence and distribution, harvest effort and sampling as per user-specified parameters. This framework can incorporate real-world heterogeneities in disease distribution, hunter harvest and harvest-based sampling, and therefore can be useful in informing wildlife disease surveillance strategies, specifically to determine population-specific sample sizes necessary for prompt detection of disease. Application of this framework is illustrated using the example of chronic wasting disease (CWD) surveillance in Missouri's white-tailed deer (Odocoileus virginianus) population. We show how confidence in detecting CWD is grossly overestimated under the unrealistic, but standard, assumptions that sampling effort and disease are randomly and independently distributed. We then provide adjusted sample size recommendations based on more realistic assumptions. Wildlife agencies can use these open-access models to design their CWD surveillance. Furthermore, these models can be readily adapted to other regions and other wildlife disease systems.

Immunosenescence in wild animals: meta-analysis and outlook.

Immunosenescence, the decline in immune defense with age, is an important mortality source in elderly humans but little is known of immunosenescence in wild animals. We systematically reviewed and meta-analysed evidence for age-related changes in immunity in captive and free-living populations of wild species (321 effect sizes in 62 studies across 44 species of mammals, birds and reptiles). As in humans, senescence was more evident in adaptive (acquired) than innate immune functions. Declines were evident for cell function (antibody response), the relative abundance of naïve immune cells and an in vivo measure of overall immune responsiveness (local response to phytohaemagglutinin injection). Inflammatory markers increased with age, similar to chronic inflammation associated with human immunosenescence. Comparisons across taxa and captive vs free-living animals were difficult due to lack of overlap in parameters and species measured. Most studies are cross-sectional, which yields biased estimates of age-effects when immune function co-varies with survival. We therefore suggest longitudinal sampling approaches, and highlight techniques from human cohort studies that can be incorporated into ecological research. We also identify avenues to address predictions from evolutionary theory and the contribution of immunosenescence to age-related increases in disease susceptibility and mortality.

Demodicosis in a captive African straw-coloured fruit bat (Eidolon helvum).

Demodicosis is most frequently observed in the domestic dog (Canis familiaris), but it has rarely been reported in bats (Chiroptera). The overpopulation of Demodex spp. that causes dermatological changes is generally associated with a compromised immune system. We describe the gross and histological features of generalized demodicosis in an adult female African straw-coloured fruit bat (Eidolon helvum) drawn from a captive research colony. The histology of the lesions revealed comedones and follicular infundubular cysts harbouring numerous Demodex spp. mites, eliciting a minimal inflammatory response in the adjacent dermis. The histological examination of a full set of tissues did not reveal clear evidence of immunosuppression, although a clinical history of recent abortion and possible stressors due to captivity could be considered risk factors for the demodicosis. Attempts to determine the Demodex species using PCR on DNA extracted from the formalin fixed paraffin embedded tissue failed. This is the first clinical and histological description of demodicosis in Eidolon helvum.

Disruption of skin microbiota contributes to salamander disease.

Escalating occurrences of emerging infectious diseases underscore the importance of understanding microbiome-pathogen interactions. The amphibian cutaneous microbiome is widely studied for its potential to mitigate disease-mediated amphibian declines. Other microbial interactions in this system, however, have been largely neglected in the context of disease outbreaks. European fire salamanders have suffered dramatic population crashes as a result of the newly emerged Batrachochytrium salamandrivorans (Bsal). In this paper, we investigate microbial interactions on multiple fronts within this system. We show that wild, healthy fire salamanders maintain complex skin microbiotas containing Bsal-inhibitory members, but these community are present at a remarkably low abundance. Through experimentation, we show that increasing bacterial densities of Bsal-inhibiting bacteria via daily addition slowed disease progression in fire salamanders. Additionally, we find that experimental-Bsal infection elicited subtle changes in the skin microbiome, with selected opportunistic bacteria increasing in relative abundance resulting in septicemic events that coincide with extensive destruction of the epidermis. These results suggest that fire salamander skin, in natural settings, maintains bacterial communities at numbers too low to confer sufficient protection against Bsal, and, in fact, the native skin microbiota can constitute a source of opportunistic bacterial pathogens that contribute to pathogenesis. By shedding light on the complex interaction between the microbiome and a lethal pathogen, these data put the interplay between skin microbiomes and a wildlife disease into a new perspective.

Long-term monitoring of an amphibian community after a climate change- and infectious disease-driven species extirpation.

Infectious disease and climate change are considered major threats to biodiversity and act as drivers behind the global amphibian decline. This is, to a large extent, based on short-term studies that are designed to detect the immediate and strongest biodiversity responses to a threatening process. What few long-term studies are available, although typically focused on single species, report outcomes that often diverge significantly from the short-term species responses. Here, we report the results of an 18-year survey of an amphibian community exposed to both climate warming and the emergence of lethal chytridiomycosis. Our study shows that the impacts of infectious disease are ongoing but restricted to two out of nine species that form the community, despite the fact all species can become infected with the fungus. Climate warming appears to be affecting four out of the nine species, but the response of three of these is an increase in abundance. Our study supports a decreasing role of infectious disease on the community, and an increasing and currently positive effect of climate warming. We caution that if the warming trends continue, the net positive effect will turn negative as amphibian breeding habitat becomes unavailable as water bodies dry, a pattern that already may be underway.

Haemoparasites of the pied flycatcher: inter-population variation in the prevalence and community composition.

The prevalence and community composition of haemoparasites can substantially differ among avian host populations, which may lead to different selection pressures. Therefore, information about these parameters is crucial for understanding, e.g. the inter-population variation in host life history traits. Here, we molecularly screened a population of a long-distance migrant, the pied flycatcher Ficedula hypoleuca, from central Poland for the presence of three genera of blood parasites: Haemoproteus, Plasmodium and Trypanosoma. The infection rate in this population was the highest for haemosporidians (86·8%) and one of the highest for trypanosomes (39·7%) among the thus far screened breeding populations of this species. The haemosporidian community was composed of six Haemoproteus/Plasmodium lineages, and the trypanosome community - 4 species and a parasite assigned to genus level. Trypanosomes were dominated by T. culicavium, a recently described species, corroborating the prediction that insectivorous songbirds are vertebrate hosts of this parasite. Host sex and age did not explain variation in infection incidence except for the higher trypanosome infection rates in males. A comparison of the study population with three other breeding populations previously screened molecularly for haemosporidians showed some geographic differences. This study confirms the importance of examining local parasite communities across a host distribution range.

High juvenile mortality in amphibians during overwintering related to fungal pathogen exposure.

The result of pathogen exposures may depend upon trade-offs in energetic demands for immune responses against host growth and survival. Environmental conditions may influence these trade-offs by affecting host size, or trade-offs may change across seasons, altering impacts of pathogens. We exposed northern leopard frog Lithobates pipiens tadpoles to different larval environments (low leaf litter, high density of conspecifics, atrazine, caged fish, or controls) that influenced size at metamorphosis. Subsequently, we exposed metamorphs to Batrachochytrium dendrobatidis (Bd), a fungal pathogen, just after metamorphosis and/or prior to overwintering 12 wk later. Bd exposure dramatically reduced survival during overwintering, with the strongest effects when hosts were exposed at both time points. Larval environments resulted in differences in host size. Those exposed to caged fish were 2.5 times larger than the smallest (those exposed to high density of conspecifics), but larval environment did not influence Bd effects on growth and survival. The largest frogs exposed to caged fish had greater survival through overwintering, but in the absence of Bd. We built stage-structured models to evaluate if overwinter mortality from Bd is capable of having effects on host populations. Our models suggest that Bd exposure after metamorphosis or before overwintering can reduce population growth rates. Our study demonstrates that hosts suffer little effects of Bd exposures following metamorphosis and that small body size did not hamper growth and survival. Instead, we provide evidence that winter mortality from Bd exposure is capable of reducing population sizes, providing a plausible mechanism for amphibian declines in temperate regions.

Haemosporidian infections in the Tengmalm's Owl (Aegolius funereus) and potential insect vectors of their transmission.

Sedentary bird species are suitable model hosts for identifying potential vectors of avian blood parasites. We studied haemosporidian infections in the Tengmalm's Owl (Aegolius funereus) in the Ore Mountains of the Czech Republic using molecular detection methods. Sex of owl nestlings was scored using molecular sexing based on fragment analysis of PCR-amplified CHD1 introns. Observed infection prevalences in nestlings and adult owls were 51 and 86 %, respectively. Five parasite lineages were detected. Most of the infections comprised the Leucocytozoon AEFUN02 and STOCC06 lineages that probably refer to distinct Leucocytozoon species. Other lineages were detected only sporadically. Mixed infections were found in 49 % of samples. The main factor affecting the probability of infection was host age. No effect of individual sex on infection probability was evidenced. The youngest infected nestling was 12 days old. High parasite prevalence in the Tengmalm's Owl nestlings suggests that insect vectors must enter nest boxes to transmit parasites before fledging. Hence, we placed sticky insect traps into modified nest boxes, collected potential insect vectors, and examined them for the presence of haemosporidian parasites using molecular detection. We trapped 201 insects which were determined as biting midges from the Culicoides genus and two black fly species, Simulium (Nevermannia) vernum and Simulium (Eusimulium) angustipes. Six haemosporidian lineages were detected in the potential insect vectors, among which the Leucocytozoon lineage BT2 was common to the Tengmalm's Owl and the trapped insects. However, we have not detected the most frequently encountered Tengmalm's Owl Leucocytozoon lineages AEFUN02 and STOCC06 in insects.

Double trouble: Co-infection of Angiostrongylus vasorum and Dirofilaria immitis in golden jackal (Canis aureus) in Friuli Venezia Giulia, Italy.

This study reports the first co-infection of Angiostrongylus vasorum and Dirofilaria immitis, two nematodes affecting canid cardiopulmonary systems, in golden jackals (Canis aureus) in Italy. Data on golden jackal carcasses, recovered in Friuli Venezia Giulia (Italy) from 2020 to 2023, were recorded using InfoFaunaFVG wildlife monitoring network. Out of 60 necropsied golden jackals, 24 tested positive for either pathogen, with 3 animals displaying co-infection. Pathological findings included verminous pneumonia, abdominal and thoracic bleeding, and adult individuals in the heart and pulmonary arteries. The recent expansion of the golden jackal in northern Italy may favour the establishment and spread of these nematodes, posing challenges for disease containment and surveillance. Given the increasing prevalence of angiostrongylosis and cardiopulmonary dirofilariosis in Europe, health monitoring of wild canids is essential to reduce their potential impact.

Development of a qPCR Duplex Assay for simultaneous detection of Fascioloides magna and Galba truncatula in eDNA samples: Monitoring beyond boundaries.

Parasites constitute a significant economic burden and highly impact environmental, public, and animal health. The emergence of many parasitic diseases is environmentally mediated and they share the same biogeography with humans and both domestic and wild animals. American liver fluke, Fascioloides magna - a trematode parasite of domestic and wild ungulates - is an example of the anthropogenic introduction of an "invasive alien species" in Italy and Europe. Multiple introductions to Europe have led to the biogeographical expansion of the parasite across the Danube region mainly provided by the presence of suitable habitats for all hosts involved in the parasite's life cycle, human-assisted transport, and drastic environmental events such as flooding. In Italy, it was introduced and established in La Mandria Regional Park (LMRP) near Turin in 1865 along with imported wapitis (Cervus elaphus canadensis) from North America (Bassi, 1875), but with no reported expansion to the surrounding areas. LMRP isolated F. magna focus, poses an important threat of possible expansion since the enclosed area is vulnerable to occasional bidirectional passage of roe deer. Additionally, tributary rivers to the Po river system, traversing the enclosed area, could further bolster the possibility of such spread. In this study, we developed a duplex qPCR assay for F. magna and its principal intermediate host Galba truncatula optimized for testing eDNA samples to meet the needs for surveillance of the parasite. Moreover, we validated the developed assay in natura by testing samples derived from filtered water and sediments collected inside and outside LMRP's fenced-off area. Our findings for the first time demonstrate the presence of F. magna's eDNA outside the park's internal fenced-off area.

Viral metagenomic survey of Caspian seals.

Introduction: Viral diseases of pinnipeds cause substantial mortality and morbidity and can influence population demography. Viral metagenomic studies can therefore play an important role in pinniped health assessments and disease surveillance relevant to both individual species and in a "One Health" context. Methods: This study used a metagenomic approach with high throughput sequencing to make the first assessment of viral diversity in Caspian seals (Pusa caspica), the only marine mammal species endemic to the Caspian Sea. Results: Sequencing libraries from 35 seals sampled 2009-2020 were analysed, finding sequences from the viral families Circoviridae, Parvoviridae, Herpesviridae, Papillomaviridae, Picornaviridae, Caliciviridae, Cruciviridae, Anelloviridae, Smacoviridae, and Orthomyxoviridae, with additional detection of Adenoviridae via PCR. The similarity of viral contigs from Caspian seal to sequences recovered from other pinnipeds ranged from 63.74% (San Miguel sea lion calicivirus) to 78.79% (Seal anellovirus 4). Discussion: Some findings represent novel viral species, but overall, the viral repertoire of Caspian seals is similar to available viromes from other pinnipeds. Among the sequences recovered were partial contigs for influenza B, representing only the second such molecular identification in marine mammals. This work provides a foundation for further studies of viral communities in Caspian seals, the diversity of viromes in pinnipeds more generally, and contributes data relevant for disease risk assessments in marine mammals.

Sustained Impact of RHDV2 on Wild Rabbit Populations across Australia Eight Years after Its Initial Detection.

Following the arrival of rabbit haemorrhagic disease virus 2 (RHDV2) in Australia, average rabbit population abundances were reduced by 60% between 2014 and 2018 based on monitoring data acquired from 18 sites across Australia. During this period, as the seropositivity to RHDV2 increased, concurrent decreases were observed in the seroprevalence of both the previously circulating RHDV1 and RCVA, a benign endemic rabbit calicivirus. However, the detection of substantial RHDV1 seropositivity in juvenile rabbits suggested that infections were continuing to occur, ruling out the rapid extinction of this variant. Here we investigate whether the co-circulation of two pathogenic RHDV variants was sustained after 2018 and whether the initially observed impact on rabbit abundance was still maintained. We monitored rabbit abundance and seropositivity to RHDV2, RHDV1 and RCVA at six of the initial eighteen sites until the summer of 2022. We observed sustained suppression of rabbit abundance at five of the six sites, with the average population reduction across all six sites being 64%. Across all sites, average RHDV2 seroprevalence remained high, reaching 60-70% in adult rabbits and 30-40% in juvenile rabbits. In contrast, average RHDV1 seroprevalence declined to <3% in adult rabbits and 5-6% in juvenile rabbits. Although seropositivity continued to be detected in a low number of juvenile rabbits, it is unlikely that RHDV1 strains now play a major role in the regulation of rabbit abundance. In contrast, RCVA seropositivity appears to be reaching an equilibrium with that of RHDV2, with RCVA seroprevalence in the preceding quarter having a strong negative effect on RHDV2 seroprevalence and vice versa, suggesting ongoing co-circulation of these variants. These findings highlight the complex interactions between different calicivirus variants in free-living rabbit populations and demonstrate the changes in interactions over the course of the RHDV2 epizootic as it has moved towards endemicity. While it is encouraging from an Australian perspective to see sustained suppression of rabbit populations in the eight years following the arrival of RHDV2, it is likely that rabbit populations will eventually recover, as has been observed with previous rabbit pathogens.

Multiple infections with H5N8 subtype high pathogenicity avian influenza viruses in a feral mallard.

During the 2020-2021 winter, Eurasian countries experienced large outbreaks caused by the clade 2.3.4.4b H5N8 subtype high pathogenicity avian influenza viruses (HPAIVs) in the wild bird populations. At least seven gene constellations have been found in the causal HPAIVs. When and where the various HPAIVs emerged remains unclear. Here, we successfully cloned H5N8 HPAIVs with multiple gene constellations from a tracheal swab of a dead mallard found at its wintering site in Japan in January 2021. According to their phylogeny, the bird was most likely co-infected with the E2 and E3 genotype clade 2.3.4.4b HPAIVs. The result indicates that feral waterbirds can be infected with multiple HPAIVs, and shed an HPAIV with novel gene constellation in Southern wintering sites.

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.

Parasite Prevalence in Feral Swine (Sus scrofa) from the Great Smoky Mountains National Park, USA.

Feral swine (Sus scrofa) are an introduced species to the Great Smoky Mountains National Park (GSMNP), US, and serve as carriers of several diseases that are considered a threat to other wildlife, domestic animals, and humans. During 2013 and 2015, fecal samples from 67 feral swine from the GSMNP within both Tennessee and North Carolina, US, were opportunistically collected as part of a feral swine removal program and submitted to the University of Tennessee College of Veterinary Medicine, Knoxville, Tennessee, for parasite screening by centrifugal sugar flotation. Ten taxa from the phyla Acanthocephala, Apicomplexa, and Nematoda were identified: Ascaris spp., Strongylid-type spp., Capillaria spp., Trichuris suis, Metastrongylus spp., Macracanthorhynchus spp., Coccidia, Sarcocystis spp., and Cryptosporidium spp. In 98.5% of samples, at least one parasite was found. No differences in parasite prevalence or species diversity were noted based on state of collection (Tennessee or North Carolina), sex, or age. The high prevalence of gastrointestinal parasites in these feral swine, some of which are zoonotic, represents a potential public health risk as well as a concern for free-range swine farmers.