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The grey wolf ( Canis lupus ) population is expanding in parts of Europe due to legal protection and favorable ecological conditions. As wolves increasingly move into urban and suburban areas, interactions with domestic dogs become more frequent, raising the risk of pathogen transmission and posing potential threats to both wolf conservation and public health. This study investigated the health status of wolves in the Foreste Casentinesi National Park (Italy) using non-invasive fecal sampling conducted between May 2019 and March 2020. Samples were genetically analyzed to identify individuals and then screened for viral pathogens, Canine Coronavirus and Parvovirus, using PCR, Sanger sequencing, and phylogenetic analysis. Parasitological examinations were performed using flotation techniques on whole samples, and real-time PCR targeting Echinococcus granulosus and E. multilocularis was conducted on selected samples. Of the 260 samples collected, genetic analysis identified 80 individual wolves belonging to 8 packs. Only one sample tested positive for Coronavirus (1.2%), and none for Parvovirus. The detected sequence clustered with strains previously reported in wolves and foxes in Italy. Copromicroscopy revealed a high prevalence of veterinary-relevant endoparasites, including Eucoleus spp. (90.0%), Sarcocystis spp. (42.5%), Taeniids (28.7%), and Ancylostomatids (26.2%). Trichuris vulpis , Toxocara canis , and coccidia showed prevalence rates below 2%. All 104 samples tested for E. granulosus or E. multilocularis were negative. These findings suggest that while wolves in the FCNP commonly harbor several canine parasites, their role in the transmission of zoonotic pathogens appears limited. Although phylogenetic data suggest that coronavirus strains tend to cluster within wildlife species, molecular data on domestic dogs remain scarce. Nonetheless, the high prevalence of shared parasites highlights the need for ongoing surveillance in both wild canids and domestic carnivores. As wolves increasingly inhabit human-dominated landscapes, understanding disease dynamics at the wildlife–domestic interface is essential for effective conservation and public health strategies.

Intensive disease surveillance in an endangered population of Ethiopian wolves provided evidence of concurrent outbreaks of rabies and canine distemper viruses in 2019, including co-infection in an individual animal. Disease surveillance and intensive monitoring of wolf packs in Ethiopia were essential in detecting the concurrent outbreaks and enabled accurate assessment of disease from both pathogens. The study highlights the risk posed to endangered populations that are susceptible to, or live in areas with, reservoir hosts for canine distemper and rabies viruses. Instances of concurrent distemper and rabies outbreaks appear unusual in the existing literature; modeling for one disease might underestimate the risk for extinction. Concurrent outbreaks may have a larger effect than single-disease outbreaks, even in a population that has partial vaccination coverage. Researchers studying wildlife populations from a conservation perspective should be aware that both diseases can strike at once where susceptible populations exist.

Wildlife reintroductions select or treat individuals for good health with the expectation that these individuals will fare better than infected animals. However, these individuals, new to their environment, may also be particularly susceptible to circulating infections and this may result in high morbidity and mortality, potentially jeopardizing the goals of recovery. Here, using the reintroduction of the grey wolf (Canis lupus) into Yellowstone National Park as a case study, we address the question of how parasites invade a reintroduced population and consider the impact of these invasions on population performance. We find that several viral parasites rapidly invaded the population inside the park, likely via spillover from resident canid species, and we contrast these with the slower invasion of sarcoptic mange, caused by the mite Sarcoptes scabiei. The spatio-temporal patterns of mange invasion were largely consistent with patterns of host connectivity and density, and we demonstrate that the area of highest resource quality, supporting the greatest density of wolves, is also the region that appears most susceptible to repeated disease invasion and parasite-induced declines. The success of wolf reintroduction appears not to have been jeopardized by infectious disease, but now shows signs of regulation or limitation modulated by parasites.

The presence of many pathogens varies in a predictable manner with latitude, with infections decreasing from the equator towards the poles. We investigated the geographic trends of pathogens infecting a widely distributed carnivore: the gray wolf ( Canis lupus ). Specifically, we investigated which variables best explain and predict geographic trends in seroprevalence across North American wolf populations and the implications of the underlying mechanisms. We compiled a large serological dataset of nearly 2000 wolves from 17 study areas, spanning 80° longitude and 50° latitude. Generalized linear mixed models were constructed to predict the probability of seropositivity of four important pathogens: canine adenovirus, herpesvirus, parvovirus, and distemper virus—and two parasites: Neospora caninum and Toxoplasma gondii . Canine adenovirus and herpesvirus were the most widely distributed pathogens, whereas N. caninum was relatively uncommon. Canine parvovirus and distemper had high annual variation, with western populations experiencing more frequent outbreaks than eastern populations. Seroprevalence of all infections increased as wolves aged, and denser wolf populations had a greater risk of exposure. Probability of exposure was positively correlated with human density, suggesting that dogs and synanthropic animals may be important pathogen reservoirs. Pathogen exposure did not appear to follow a latitudinal gradient, with the exception of N. caninum . Instead, clustered study areas were more similar: wolves from the Great Lakes region had lower odds of exposure to the viruses, but higher odds of exposure to N. caninum and T. gondii ; the opposite was true for wolves from the central Rocky Mountains. Overall, mechanistic predictors were more informative of seroprevalence trends than latitude and longitude. Individual host characteristics as well as inherent features of ecosystems determined pathogen exposure risk on a large scale. This work emphasizes the importance of biogeographic wildlife surveillance, and we expound upon avenues of future research of cross-species transmission, spillover, and spatial variation in pathogen infection.

Aujeszky’s disease (AD), caused by suid herpesvirus 1 (pseudorabies virus, PRV), is a highly contagious infection primarily affecting swine, with wild boars serving as an important reservoir in Europe. Spillover infections in non-suid species, including carnivores, are rare but typically fatal and of epidemiological concern. This study presents the first case of AD in a grey wolf (Canis lupus) in Central Europe with genomic characterization. The 8-month-old wolf was found in the Carpathians (SE Poland), moribund with acute neurological signs, and euthanized for animal welfare reasons. Necropsy revealed no pathognomonic gross lesions. Molecular analyses of tissues confirmed the presence of PRV DNA using real-time PCR, and virus isolation was successful. Genomic analysis revealed that the PRV isolate clustered within genotype I, the predominant circulating genotype in Europe. However, due to the limited availability of reference PRV genome sequences from European wildlife, the precise geographic origin and transmission pathways of this strain could not be fully resolved. In the presented case, wild boars were considered a possible source of infection. This highlights the potential for PRV transmission to apex predators. This study emphasizes the importance of systematic surveillance of PRV in wildlife and the need for expanded genomic databases of PRV strains. Full-genome sequencing is crucial for improving the understanding of PRV transmission.

Novel protoparvoviruses genetically related to human and non-human primate bufaviruses (BuVs) have been detected recently in respiratory and enteric specimens collected from dogs and cats. In this study, by molecular screening of archival collections of faecal samples from wolves and foxes, we detected BuVs with a rate of 17.1% (7/41) and 10.5% (9/86), respectively. Sequence analysis of a portion of the ORF2 gene region of nine positive samples showed that the viruses in these samples were closely related to BuVs (97.5–99.0% nucleotide sequence identity) found in domestic carnivores.

We provide the first report of a wolf infected with pseudorabies virus (PRV) in China. We observed the clinical symptoms and also dissected tissue samples from the wolf. The samples were ground under sterile conditions and injected subcutaneously into the necks of rabbits, which subsequently developed intense pruritus symptoms and died. The PRV strain from the wolf was isolated in porcine kidney (PK)-15 cells and was specifically recognized by pig PRV antibody-positive serum, as shown by indirect immunofluorescence. Tissues from the dead wolf and rabbits were examined by polymerase chain reaction (PCR), and the PCR-amplified partial glycoprotein E gene was sequenced, which confirmed that the wolf had died as a result of PRV infection.

Canine kobuviruses (CaKoVs) were first identified in diarrhoeic and asymptomatic dogs in 2011 in the USA. Subsequent studies have demonstrated a worldwide distribution of these viruses, but it is not clear if CaKoVs play a role as enteric pathogens of dogs. More recently, CaKoV RNA has been detected in wild carnivores, including red fox, golden jackal, side-striped jackal and spotted hyena. In this study, we addressed the hypothesis that wolves are susceptible to CaKoV infections. A total of 185 wolf stool samples were collected from necropsied animals and from transects in the Liguria, Piemonte and Valle D’Aosta regions of Italy, and CaKoV RNA was identified in two of these specimens. Both samples were obtained from necropsied wolves, with a prevalence rate of 4.9% (2/41). Sequence analysis of the full-length VP1 region showed that these strains displayed the highest nucleotide (nt) sequence identity (86.3-98.5%) to canine strains identified in the UK and Africa, and to kobuviruses that were previously detected in other African wild carnivores. This suggests that genetically related CaKoV strains circulate in domestic and wild carnivores, with interspecies transmission being not uncommon among carnivores of different ecosystems.

African swine fever (ASF), caused by a DNA virus (ASFV) belonging to genus Asfivirus of the Asfarviridae family, is one of the most threatening diseases of suids. During last few years, it has spread among populations of wild boars and pigs in countries of Eastern and Central Europe, causing huge economical losses. While local ASF occurrence is positively correlated with wild boar density, ecology of this species (social structure, movement behavior) constrains long-range disease transmission. Thus, it has been speculated that carnivores known for high daily movement and long-range dispersal ability, such as the wolf (Canis lupus), may be indirect ASFV vectors. To test this, we analyzed 62 wolf fecal samples for the presence of ASFV DNA, collected mostly in parts of Poland declared as ASF zones. This dataset included 20 samples confirmed to contain wild boar remains, 13 of which were collected near places where GPS-collared wolves fed on dead wild boars. All analyzed fecal samples were ASFV-negative. On the other hand, eight out of nine wild boar carcasses that were fed on by telemetrically studied wolves were positive. Thus, our results suggest that when wolves consume meat of ASFV-positive wild boars, the virus does not survive the passage through intestinal tract. Additionally, wolves may limit ASFV transmission by removing infectious carrion. We speculate that in areas where telemetric studies on large carnivores are performed, data from GPS collars could be used to enhance efficiency of carcass search, which is one of the main preventive measures to constrain ASF spread.

Canine distemper virus (CDV) infection is a primary threat affecting a wide number of carnivore species, including wild animals. In January 2013, two carcasses of Apennine wolves (Canis lupus) were collected in Ortona dei Marsi (L'Aquila province, Italy) by the local Veterinary Services. CDV was immediately identified either by RT-PCR or immunohistochemistry in lung and central nervous tissue samples. At the same time, severe clinical signs consistent with CDV infection were identified and taped (Videos S1-S3) from three wolves rescued in the areas surrounding the National Parks of the Abruzzi region by the Veterinary Services. The samples collected from these symptomatic animals also turned out CDV positive by RT-PCR. So far, 30 carcasses of wolves were screened and CDV was detected in 20 of them. The sequencing of the haemagglutinin gene and subsequent phylogenetic analysis demonstrated that the identified virus belonged to the CDV Arctic lineage. Strains belonging to this lineage are known to circulate in Italy and in Eastern Europe amongst domestic dogs. To the best of our knowledge this is the first report of CDV Arctic lineage epidemics in the wild population in Europe.