SARS-CoV-2 Seropositivity in Urban Population of Wild Fallow Deer, Dublin, Ireland, 2020-2022.

SARS-CoV-2 can infect wildlife, and SARS-CoV-2 variants of concern might expand into novel animal reservoirs, potentially by reverse zoonosis. White-tailed deer and mule deer of North America are the only deer species in which SARS-CoV-2 has been documented, raising the question of whether other reservoir species exist. We report cases of SARS-CoV-2 seropositivity in a fallow deer population located in Dublin, Ireland. Sampled deer were seronegative in 2020 when the Alpha variant was circulating in humans, 1 deer was seropositive for the Delta variant in 2021, and 12/21 (57%) sampled deer were seropositive for the Omicron variant in 2022, suggesting host tropism expansion as new variants emerged in humans. Omicron BA.1 was capable of infecting fallow deer lung type-2 pneumocytes and type-1-like pneumocytes or endothelial cells ex vivo. Ongoing surveillance to identify novel SARS-CoV-2 reservoirs is needed to prevent public health risks during human-animal interactions in periurban settings.

Case report: First documented case of cerebral angiostrongyliasis caused by Angiostrongylus costaricensis in a free-ranging opossum.

Angiostrongylus costaricensis is a metastrongyloid nematode that primarily infects the mesenteric arteries of wild rodents. This parasite is endemic in several regions of the American continent, and in humans, causes a disease known as abdominal angiostrongyliasis. Despite the important health implications of this nematode, there are limited studies investigating the involvement of wild animals in its life cycle. In this study, we present the clinical manifestations, pathologic findings, and molecular diagnosis, to the best of our current knowledge, of the first documented onset of cerebral angiostrongyliasis because of A. costaricensis infection in a juvenile free-ranging opossum (Didelphis marsupialis). Histopathological findings stress the presence of eosinophilic meningoencephalitis with nematodes present within the lesions, and PCR was positive for cox1 and ITS1 reactions. The obtained sequences for a 279 bp fragment of ITS1 were 100% identical to A. costaricensis from Costa Rica. This case highlights the substantial difficulties in diagnosing neuroangiostrongyliasis, yet underscores the importance of considering A. costaricensis as a potential culprit behind neurological conditions in wild marsupials. It acts as an urgent call to action to improve surveillance programs tracking infectious and parasitic diseases causing mortality in wildlife populations.

Is the European badger a new host for Dirofilaria immitis? The first records in Greece.

Dirofilaria immitis is a ubiquitous nematode parasite with zoonotic potential, transmitted by mosquitoes, that causes heartworm disease in various animal species. Dogs are the parasite's typical final host, and wild carnivores represent the parasite's reservoir in nature. Studies on D. immitis infections in wild animals are essential to assess infection pressure for domestic animals, and until now, there has been only one infection case reported in a European badger (Meles meles). The current report describes the first two European badger cases with cardiovascular dirofilariosis in Greece. Two adult male badgers were rescued in Heraklion and Chania, Crete Island, and admitted to "ANIMA -Wildlife Rehabilitation Centre" in Athens. The detailed clinical examination revealed that the first badger suffered from severe broncho-pneumonitis while the second one displayed clinical signs associated with severe brain trauma. Blood samples were taken for haematology and biochemistry analyses during their short hospitalisation period. In addition, different routine diagnostic tests were carried out, including heartworm antigen testing (ELISA) and the modified Knott's test for microfilariae. Both badgers were positive in both tests. The animals died a few hours after their admission and the detailed necropsies followed, revealed the presence of three parasites in each animal's right heart, morphologically identified as adults of D. immitis. These findings add the European badger in the list of additional potential reservoir hosts for D. immitis and highlight the potential role of wildlife for companion animals and human health.

White-tailed deer (Odocoileus virginianus) may serve as a wildlife reservoir for nearly extinct SARS-CoV-2 variants of concern.

The spillover of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from humans to white-tailed deer (WTD) and its ability to transmit from deer to deer raised concerns about the role of WTD in the epidemiology and ecology of the virus. Here, we present a comprehensive cross-sectional study assessing the prevalence, genetic diversity, and evolution of SARS-CoV-2 in WTD in the State of New York (NY). A total of 5,462 retropharyngeal lymph node samples collected from free-ranging hunter-harvested WTD during the hunting seasons of 2020 (Season 1, September to December 2020, n = 2,700) and 2021 (Season 2, September to December 2021, n = 2,762) were tested by SARS-CoV-2 real-time RT-PCR (rRT-PCR). SARS-CoV-2 RNA was detected in 17 samples (0.6%) from Season 1 and in 583 samples (21.1%) from Season 2. Hotspots of infection were identified in multiple confined geographic areas of NY. Sequence analysis of SARS-CoV-2 genomes from 164 samples demonstrated the presence of multiple SARS-CoV-2 lineages and the cocirculation of three major variants of concern (VOCs) (Alpha, Gamma, and Delta) in WTD. Our analysis suggests the occurrence of multiple spillover events (human to deer) of the Alpha and Delta lineages with subsequent deer-to-deer transmission and adaptation of the viruses. Detection of Alpha and Gamma variants in WTD long after their broad circulation in humans in NY suggests that WTD may serve as a wildlife reservoir for VOCs no longer circulating in humans. Thus, implementation of continuous surveillance programs to monitor SARS-CoV-2 dynamics in WTD is warranted, and measures to minimize virus transmission between humans and animals are urgently needed.

Detection of Dirofilaria immitis via integrated serological and molecular analyses in coyotes from Texas, United States.

Wild canids serve as reservoir for various vector-borne pathogens of veterinary and medical importance, including the canine heartworm, Dirofilaria immitis. In North and Central America, coyotes (Canis latrans) may be a relevant reservoir host for heartworm transmission. The objective of this study was to determine the occurrence of D. immitis in coyotes across Texas using integrated antigen detection test and molecular assays. Matching whole blood and serum samples were collected from 122 coyotes from different locations across the state of Texas, United States, encompassing nine counties. Collections occurred from February to April 2016, and December 2016. Samples were assessed serologically using a commercial microtiter plate ELISA (DiroCHEK®), and molecularly by conventional PCR targeting the cytochrome oxidase c subunit 1 (cox1) and NADH dehydrogenase subunit 5 (nad5) of the mitochondrial DNA, and via a TaqMan© probe-based real-time PCR protocol, also targeting a fragment of the cox1 gene. Overall, 12 (9.83%) samples tested positive when serological and molecular results were combined. Seven of 122 samples (5.73%) were antigen-positive, 8 (6.55%) were qPCR-positive, and 4 (3.27%) were positive using conventional PCR. Of 12 positive samples, 4 tested antigen-positive by DiroCHEK® but were negative in all molecular tests, another 4 tested positive by at least one of the molecular assays but tested negative by DiroCHEK®, and 3 samples tested positive by both antigen test and at least one of the molecular assays. Two samples (16.67%) tested positive on both the antigen test and both conventional PCR and qPCR. Our study confirmed the presence of D. immitis infection in coyotes from southern and northern Texas. The combination of serologic and molecular diagnostic tests was proven synergistic for the identification of D. immitis infections, including occult dirofilariosis, and revealed a more accurate picture of heartworm occurrence in the sampled coyotes.

Is serology a realistic approach for monitoring red deer tuberculosis in the field?

Tuberculosis (TB) is a zoonotic mycobacterial infection with great importance in human health, animal production, and wildlife conservation. Although an ambitious eradication programme in cattle has been implemented for decades, TB-free status has not yet been achieved in most of Spain, where animal TB persists in a multi-host system of domestic and wild hosts, including the red deer (Cervus elaphus). However, information on long time series and trends of TB prevalence in wildlife is scarce. The diagnosis of TB in wild red deer is often based on gross pathology and bacteriological culture confirmation, although recently serological assays have been developed to detect anti- Mycobacterium tuberculosis Complex (MTC) antibodies. Particularly, protein complex P22 has demonstrated to yield good specificity and sensitivity in the serological diagnosis of MTC for red deer, as well as cattle, goats, sheep, pigs, wild boar, and European badger. Thus, the objective of the present study was to compare the performance of the P22-ELISA with TB-compatible lesion detection, as well as to assess the potential application of each technique for determining spatiotemporal trends and risk factors of MTC infection in wild red deer from low and high TB prevalence areas of Spain over the last two decades. We tested 5095 sera from 13 wild populations by indirect ELISA using P22 as antigen. Mean seroprevalence (13.22%, CI95: 12.32-14.18) was compared with the prevalence of macroscopic TB-compatible lesions (6.94%, CI95: 6.18-7.79). The results evidenced a poor agreement between both techniques (K < 0.3), although generalized TB-lesions and anti-P22 antibodies showed a positive association (χ² = 9.054, P = 0.004). Consequently, TB-lesion based prevalence and seroprevalence cannot be considered as equivalent for TB surveillance in red deer. Regarding the spatiotemporal trend of TB in red deer in Spain, we observed a North-South gradient of TB occurrence [North: 1.23% (CI95: 0.77-1.97) of TB-lesions and 12.55% (CI95: 10.91-14.41) of P22-ELISA; Centre: 7.10% (CI95: 6.04-8.33) and 8.74% (CI95: 7.57-10.08); South: 21.04% (CI95:17.81-24.69) and 23.09% (CI95: 19.73-26.84), respectively]. Overall, there was a stability over time, with higher prevalence in adults belonging to densely populated sites. We conclude that the P22-ELISA alone is not sufficiently reliable for TB surveillance in red deer at large spatiotemporal scales. Instead, we recommend combining gross pathology and P22-ELISA.

The Importance of Accurate Host Species Identification in the Framework of Rabies Surveillance, Control and Elimination.

Accurate host identification is paramount to understand disease epidemiology and to apply appropriate control measures. This is especially important for multi-host pathogens such as the rabies virus, a major and almost invariably fatal zoonosis that has mobilized unanimous engagement at an international level towards the final goal of zero human deaths due to canine rabies. Currently, diagnostic laboratories implement a standardized identification using taxonomic keys. However, this method is challenged by high and undiscovered biodiversity, decomposition of carcasses and subjective misevaluation, as has been attested to by findings from a cohort of 242 archived specimens collected across Sub-Saharan Africa and submitted for rabies diagnosis. We applied two simple and cheap methods targeting the Cytochrome b and Cytochrome c oxidase subunit I to confirm the initial classification. We therefore suggest prioritizing a standardized protocol that includes, as a first step, the implementation of taxonomic keys at a family or subfamily level, followed by the molecular characterization of the host species.

SARS-CoV-2 in animals: potential for unknown reservoir hosts and public health implications.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, previously 2019-nCoV) is suspected of having originated in 2019 in China from a coronavirus infected bat of the genus Rhinolophus. Following the initial emergence, possibly facilitated by a mammalian bridge host, SARS-CoV-2 is currently transmitted across the globe via efficient human-to-human transmission. Results obtained from experimental studies indicate that animal species such as cats, ferrets, raccoon dogs, cynomolgus macaques, rhesus macaques, white-tailed deer, rabbits, Egyptian fruit bats, and Syrian hamsters are susceptible to SARS-CoV-2 infection, and that cat-to-cat and ferret-to-ferret transmission can take place via contact and air. However, natural infections of SARS-CoV-2 have been reported only in pet dogs and cats, tigers, lions, snow leopards, pumas, and gorillas at zoos, and farmed mink and ferrets. Even though human-to-animal spillover has been reported at several instances, SARS-CoV-2 transmission from animals-to-humans has only been reported from mink-to-humans in mink farms. Following the rapid transmission of SARS-CoV-2 within the mink population, a new mink-associated SARS-CoV-2 variant emerged that was identified in both humans and mink. The increasing reports of SARS-CoV-2 in carnivores indicate the higher susceptibility of animal species belonging to this order. The sporadic reports of SARS-CoV-2 infection in domestic and wild animal species require further investigation to determine if SARS-CoV-2 or related Betacoronaviruses can get established in kept, feral or wild animal populations, which may eventually act as viral reservoirs. This review analyzes the current evidence of SARS-CoV-2 natural infection in domestic and wild animal species and their possible implications on public health.

New serological platform for detecting antibodies against Mycobacterium tuberculosis complex in European badgers.

European badgers (Meles meles) have been identified as wildlife reservoirs for Mycobacterium bovis in the UK and Ireland, and may also have a role in the epidemiology of animal tuberculosis in other European regions. Thus, detection of M. bovis-infected badgers may be required for the purposes of surveillance and monitoring of disease levels in infected populations. Current serological assays to detect M. bovis infection in live badgers, while rapid and inexpensive, show limited diagnostic sensitivity. Here we describe and evaluate new ELISA platforms for the recognition of the P22 multiprotein complex derived from the purified protein derivative (PPD) of M. bovis. The recognition of IgG against P22 multiprotein complex derived from PPD-B was tested by ELISA in the serum of badgers from the UK, Ireland and Spain. TB infection in the badgers was indicated by the presence of M. bovis in tissues by culture and histology at post-mortem examination and TB-free status was established by repeated negativity in the interferon γ release assay (IGRA). In experimentally infected badgers, humoral antibody responses against P22 developed within 45 days post-infection. The ELISA tests showed estimated sensitivity levels of 74-82% in experimentally and naturally infected badgers with specificities ranging from 75% to 100% depending on the badger population tested. The P22 multi-antigen based ELISAs provide a sensitive and specific test platform for improved tuberculosis surveillance in badgers.

A new bovine tuberculosis model for England and Wales (BoTMEW) to simulate epidemiology, surveillance and control.

BACKGROUND: Bovine tuberculosis (bTB) is a zoonotic disease of cattle caused by Mycobacterium bovis, widespread in England and Wales. It has high incidence towards the South West of England and Wales, with much lower incidence to the East and North. A stochastic simulation model was developed to simulate M. bovis transmission among cattle, transfer by cattle movements and transmission from environmental reservoirs (often wildlife and especially badgers). It distinguishes five surveillance streams, including herd tests, pre-movement testing and slaughter surveillance. The model thereby simulates interventions in bTB surveillance and control, and generates outputs directly comparable to detailed disease records. An anonymized version of the executable model with its input data has been released. The model was fitted to cattle bTB records for 2008-2010 in a cross-sectional comparison, and its projection was compared with records from 2010 to 2016 for validation. RESULTS: The fitted model explained over 99% of the variation among numbers of breakdowns in four defined regions and surveillance streams in 2010. It classified 7800 (95% confidence interval c. 5500 - 14,000) holdings within high incidence regions as exposed to infectious environmental reservoirs, out of over 31,000 cattle holdings identified as potentially exposed to such sources. The model was consistent with previous estimates of low M. bovis transmission rate among cattle, but cattle to cattle transmission was clearly required to generate the number of cattle cases observed. When projected to 2016, the model as fitted to 2010 continued to match the distribution of bTB among counties, although it was notable that the actual distribution of bTB in 2010 was itself a close match for its distribution in 2016. CONCLUSIONS: The close model fit demonstrated that cattle movements could generate breakdowns as observed in low incidence regions, if persistent environmental reservoirs such as wildlife maintained infection levels in the high incidence regions. The model suggests that environmental reservoirs may be a challenge for control, because, although many holdings are exposed to infection from wildlife or the environment, they are a minority of holdings. Large impacts on disease in wildlife will be required to avoid each individual transmission event to cattle.

Effects of a zoonotic pathogen, Borrelia burgdorferi, on the behavior of a key reservoir host.

Most emerging infectious diseases of humans are transmitted to humans from other animals. The transmission of these "zoonotic" pathogens is affected by the abundance and behavior of their wildlife hosts. However, the effects of infection with zoonotic pathogens on behavior of wildlife hosts, particularly those that might propagate through ecological communities, are not well understood. Borrelia burgdorferi is a bacterium that causes Lyme disease, the most common vector-borne disease in the USA and Europe. In its North American range, the pathogen is most frequently transmitted among hosts through the bite of infected blacklegged ticks (Ixodes scapularis). Using sham and true vaccines, we experimentally manipulated infection load with this zoonotic pathogen in its most competent wildlife reservoir host, the white-footed mouse, Peromyscus leucopus, and quantified the effects of infection on mouse foraging behavior, as well as levels of mouse infestation with ticks. Mice treated with the true vaccine had 20% fewer larval blacklegged ticks infesting them compared to mice treated with the sham vaccine, a significant difference. We observed a nonsignificant trend for mice treated with the true vaccine to be more likely to visit experimental foraging trays (20%-30% effect size) and to prey on gypsy moth pupae (5%-20% effect size) compared to mice treated with the sham vaccine. We observed no difference between mice on true- versus sham-vaccinated grids in risk-averse foraging. Infection with this zoonotic pathogen appears to elicit behavioral changes that might reduce self-grooming, but other behaviors were affected subtly or not at all. High titers of B. burgdorferi in mice could elicit a self-reinforcing feedback loop in which reduced grooming increases tick burdens and hence exposure to tick-borne pathogens.

Interspecific network centrality, host range and early-life development are associated with wildlife hosts of Rift Valley fever virus.

Rift Valley fever virus (RVFV) is responsible for a substantive disease burden in pastoralist communities and the agricultural sector in the African continent and Arabian Peninsula. Enzootic, epizootic and zoonotic RVFV transmission dynamics remain ill-defined, particularly due to a poor understanding of the role of mammalian hosts in the epidemiology and infection ecology of this arbovirus. Using a piecewise structural equation model, this study sought to identify associations between biological and ecological characteristics of mammalian species and documented RVFV infection to highlight species-level traits that may influence wildlife host status. Interspecific network centrality, size of species home range and reproductive life-history traits were all associated with being an RVFV host. The identification of these species-level characteristics may help to provide ecological context for the role of wildlife amplification hosts in the epidemiology of spillover to livestock and humans and may also help to identify specific points of vulnerability at the wildlife-livestock interface.

Interannual cycles of Hantaan virus outbreaks at the human-animal interface in Central China are controlled by temperature and rainfall.

Hantavirus, a rodent-borne zoonotic pathogen, has a global distribution with 200,000 human infections diagnosed annually. In recent decades, repeated outbreaks of hantavirus infections have been reported in Eurasia and America. These outbreaks have led to public concern and an interest in understanding the underlying biological mechanisms. Here, we propose a climate-animal-Hantaan virus (HTNV) infection model to address this issue, using a unique dataset spanning a 54-y period (1960-2013). This dataset comes from Central China, a focal point for natural HTNV infection, and includes both field surveillance and an epidemiological record. We reveal that the 8-y cycle of HTNV outbreaks is driven by the confluence of the cyclic dynamics of striped field mouse (Apodemus agrarius) populations and climate variability, at both seasonal and interannual cycles. Two climatic variables play key roles in the ecology of the HTNV system: temperature and rainfall. These variables account for the dynamics in the host reservoir system and markedly affect both the rate of transmission and the potential risk of outbreaks. Our results suggest that outbreaks of HTNV infection occur only when climatic conditions are favorable for both rodent population growth and virus transmission. These findings improve our understanding of how climate drives the periodic reemergence of zoonotic disease outbreaks over long timescales.

Cattle-derived Salmonella enterica serovar Dublin Infections in Red Foxes ( Vulpes vulpes ) in Tyrol, Austria.

Salmonella enterica serovar Dublin is endemic in the cattle population in some areas of the Austrian province Tyrol, and each year single dairy farms have experienced clinical infections. To ascertain if Tyrolean red foxes ( Vulpes vulpes ) act as a reservoir for Salmonella spp., we tested hepatic tissue and intestinal content from foxes hunted in the years 2015-16 by using microbiological methods. In addition, we included several fox fecal samples collected on a mountain pasture near chamois carcasses in the investigation. Of 434 foxes tested, nine animals (2.1%) were positive for Salmonella spp. Serotyping revealed five foxes positive with S. Dublin, demonstrating that this serovar exists in the Tyrolean fox population. The fecal samples collected in the area surrounding skeletonized chamois ( Rupicapra rupicapra ) also tested positive for S. Dublin. These chamois were probably victims of a waterborne outbreak caused by S. Dublin-shedding cattle. Our results indicate that the S. Dublin infections in red foxes were primarily acquired through ingestion of infected cattle material such as abortion tissues, but also by feeding on dead chamois. The findings underline the importance of interspecies transmission in this domestic/wildlife interface.

Parenteral Vaccination with Heat-Inactivated Mycobacterium Bovis Reduces the Prevalence of Tuberculosis-Compatible Lesions in Farmed Wild Boar.

In 2012, a wild boar (Sus scrofa) tuberculosis (TB) control programme was set up in a wild boar farm by means of intramuscular (IM) vaccination with a heat-inactivated Mycobacterium bovis vaccine (IV). The goal was to assess safety and efficacy of the parenterally administered IV in a large farm setting with natural M. bovis circulation. Based on preceding results under laboratory conditions, we hypothesized that vaccinated piglets would show smaller scores of TB-compatible lesions (TBCL) than unvaccinated controls. After vaccination, no adverse reactions were detected by visual inspection or at post-mortem examination (n = 668 and 97, respectively). Post-mortem data on TBCL were available for 97 vaccinated wild boar and 182 controls. The observed TBCL prevalence was 4.1% (95% CI = 0.2-8%) and 12.1% (95% CI = 7.1-17.1%) for vaccinated and control wild boar, respectively (P < 0.05). Among those animals with TBCL, no difference in the mean lesion score was found (P > 0.05). The results show that IV administered intramuscularly to wild boar piglets is safe and protects vaccinated individuals (66% reduction in TBCL prevalence) against natural challenge in a low-prevalence setting. In a context of increasing TB prevalence in wild boar in Mediterranean habitats, vaccination achieved a progressive though slow decline in lesion prevalence since the onset of the vaccination scheme. Hence, vaccination might contribute, along with other tools, to TB control in wild boar and in pigs.

Bovine Viral Diarrhea Virus (BVDV) in White-Tailed Deer (Odocoileus virginianus).

Bovine viral diarrhea virus (BVDV) is the prototypic member of the genus Pestivirus in the family Flaviviridae. Infections with BVDV cause substantial economic losses to the cattle industries, prompting various organized control programs in several countries. In North America, these control programs are focused on the identification and removal of persistently infected (PI) cattle, enhancement of BVDV-specific immunity through vaccination, and the implementation of biosecure farming practices. To be successful, control measures must be based on complete knowledge of the epidemiology of BVDV, including the recognition of other potential sources of the virus. BVDV does not possess strict host-specificity, and infections of over 50 species in the mammalian order Artiodactyla have been reported. Over 50 years ago, serologic surveys first suggested the susceptibility of white-tailed deer (Odocoileus virginianus), the most abundant free-ranging ruminant in North America, to BVDV. However, susceptibility of white-tailed deer to BVDV infection does not alone imply a role in the epidemiology of the virus. To be a potential wildlife reservoir, white-tailed deer must: (1) be susceptible to BVDV, (2) shed BVDV, (3) maintain BVDV in the population, and (4) have sufficient contact with cattle that allow spillback infections. Based on the current literature, this review discusses the potential of white-tailed deer to be a reservoir for BVDV.

Demographic buffering and compensatory recruitment promotes the persistence of disease in a wildlife population.

Demographic buffering allows populations to persist by compensating for fluctuations in vital rates, including disease-induced mortality. Using long-term data on a badger (Meles meles Linnaeus, 1758) population naturally infected with Mycobacterium bovis, we built an integrated population model to quantify impacts of disease, density and environmental drivers on survival and recruitment. Badgers exhibit a slow life-history strategy, having high rates of adult survival with low variance, and low but variable rates of recruitment. Recruitment exhibited strong negative density-dependence, but was not influenced by disease, while adult survival was density independent but declined with increasing prevalence of diseased individuals. Given that reproductive success is not depressed by disease prevalence, density-dependent recruitment of cubs is likely to compensate for disease-induced mortality. This combination of slow life history and compensatory recruitment promotes the persistence of a naturally infected badger population and helps to explain the badger's role as a persistent reservoir of M. bovis.

Comparison of Rift Valley fever virus replication in North American livestock and wildlife cell lines.

Rift Valley fever virus (RVFV) causes disease outbreaks across Africa and the Arabian Peninsula, resulting in high morbidity and mortality among young domestic livestock, frequent abortions in pregnant animals, and potentially severe or fatal disease in humans. The possibility of RVFV spreading to the United States or other countries worldwide is of significant concern to animal and public health, livestock production, and trade. The mechanism for persistence of RVFV during inter-epidemic periods may be through mosquito transovarial transmission and/or by means of a wildlife reservoir. Field investigations in endemic areas and previous in vivo studies have demonstrated that RVFV can infect a wide range of animals, including indigenous wild ruminants of Africa. Yet no predominant wildlife reservoir has been identified, and gaps in our knowledge of RVFV permissive hosts still remain. In North America, domestic goats, sheep, and cattle are susceptible hosts for RVFV and several competent vectors exist. Wild ruminants such as deer might serve as a virus reservoir and given their abundance, wide distribution, and overlap with livestock farms and human populated areas could represent an important risk factor. The objective of this study was to assess a variety of cell lines derived from North American livestock and wildlife for susceptibility and permissiveness to RVFV. Results of this study suggest that RVFV could potentially replicate in native deer species such as white-tailed deer, and possibly a wide range of non-ruminant animals. This work serves to guide and support future animal model studies and risk model assessment regarding this high-consequence zoonotic pathogen.

Molecular identification of Ehrlichia species and host bloodmeal source in Amblyomma americanum L. from two locations in Tennessee, United States.

The current status of tick-borne diseases in the southeastern United States is challenging to define due to emerging pathogens, uncertain tick/host relationships, and changing disease case definitions. A golf-oriented retirement community on the Cumberland Plateau in Tennessee experienced an ehrlichiosis outbreak in 1993, prompting efforts to reduce the local tick population using '4-Poster' acaricide devices targeting white-tailed deer (Odocoileus virginianus). In 2009, the prevalence of Ehrlichia spp. in questing ticks was surveyed in the area and compared to a Tennessee state park where acaricide had not been applied. The range of wildlife hosts that immature Amblyomma americanum fed upon and the role that these hosts may play in pathogen dynamics were investigated using a reverse line blot (RLB) bloodmeal analysis technique. Amblyomma americanum was by far the most common tick species in both study areas (>99% of ticks collected). Of 303 adult and nymphal A. americanum tested at the retirement community, six were positive for Ehrlichia chaffeensis (2.0%), 16 were positive for E. ewingii (5.3%), and six were positive for Panola Mountain Ehrlichia (2.0%). This is the first confirmation of Panola Mountain Ehrlichia in A. americanum from the state of Tennessee. The 9.3% prevalence of Ehrlichia spp. in ticks from the retirement community was similar to that detected at the state park site (5.5%), suggesting that the 4-Poster treatment had not been sufficient to reduce Ehrlichia spp. cycling in the tick population. At both study sites, A. americanum fed on a wide range of mammal and bird species, with a minority of detectable bloodmeals coming from deer. Of the Ehrlichia-infected nymphs with positive bloodmeal identification, none fed on deer, indicating that multiple vertebrate species are contributing to sylvatic maintenance of Ehrlichia spp. at these sites. This highlights the difficulty of attempting to reduce the risk of tick-borne disease through host-targeted interventions alone.

Seroprevalence of Encephalitozoon cuniculi in wild rodents, foxes and domestic cats in three sites in the United Kingdom.

Encephalitozoon cuniculi is an obligate intracellular microsporidian that is the causal agent of encephalitozoonosis, an important and emerging disease in both humans and animals. Little is known about its occurrence in wildlife. In this study, serum samples from 793 wild rodents [178 bank voles (BV), 312 field voles (FV) and 303 wood mice (WM)], 96 foxes and 27 domestic cats from three study areas in the UK were tested for the presence of antibodies to E. cuniculi using a direct agglutination test (DAT). Seroprevalence in the wild rodents ranged from 1.00% to 10.67% depending on species (overall 5.31%) and was significantly higher in foxes [49.50% (50/96)]. None of the 27 cats sampled were found to be seropositive. This is the first report of seroprevalence to E. cuniculi in BV, FV, WM, foxes and cats in the UK and provides some evidence that foxes could act as sentinels for the presence of E. cuniculi in rodents. The study demonstrates that wildlife species could be significant reservoirs of infection for both domestic animals and humans.