Free-ranging Wolves Research Articles

Monitoring of pathogenic Leptospira infection in wolves (Canis lupus) from Spain and Italy

Leptospirosis is a bacterial disease of worldwide distribution with relevant implications for animal and human health. Different large wild carnivore species can act as reservoirs of this zoonotic pathogen. This study aimed to evaluate the circulation of Leptospira spp. in free-ranging wolves (Canis lupus) from southern Europe. A total of 281 kidney samples of wolves from Spain and Italy were collected between 2017 and 2023. The presence of Leptospira DNA was analysed by real-time PCR and phylogenetic analyses were carried out using a Bayesian approach. The overall prevalence was 3.2 % (9/281; 95 %CI: 1.1–5.3). Leptospira DNA was detected in nine of the 180 wolves from Spain (5.0 %; 95 %CI: 1.8–8.2), but not in the Italian wolf population (0 %; 0/101). Molecular analyses revealed high homology between the sequences obtained in the present study and isolates of Leptospira interrogans and Leptospira borgpetersenii from different rodent and domestic ungulate species. Our results provide evidence of a low and spatially heterogeneous circulation of this pathogen in wolf populations of southern Europe. The detection of zoonotic Leptospira species in this survey supports the need to consider wolf populations in monitoring programs for leptospirosis with a One Health approach.

Epidemiological survey of infectious agents in free-ranging maned wolves (Chrysocyon brachyurus) in Northeastern Brazil.

Infectious diseases are one of the most concerning threats to maned wolves (Chrysocyon brachyurus) due to the potential impact on free-ranging populations. The species is currently classified as vulnerable according to the national list of threatened species and occurs mainly in open habitats, such as the Cerrado, a tropical savannah, which comprises its main distribution area in Brazil. In the northeastern region, it occurs in the Cerrado of Bahia, Piauí, Maranhão, and Tocantins states. Therefore, this study aimed to investigate the occurrence of infectious agents in Chrysocyon brachyurus through an epidemiological assessment of free-ranging individuals in western Bahia, specifically in the Barreiras microregion, a Cerrado area intensely fragmented and anthropized by agricultural activity. Eleven specimens were evaluated for serological titration, antigen research, and genetic material research for canine distemper virus (CDV), canine parvovirus (CPV), adenovirus-canine-type 1 (CAdV-1), canine coronavirus (CCoV), Leptospira interrogans and Toxoplasma gondii from 2020 to 2022. In addition to maned wolves, domestic dogs were also evaluated and tested. All maned wolves (100%) evaluated by the dot-ELISA technique exhibited immunoglobulin M (IgM) and seven (64%) exhibited immunoglobulin G (IgG) against CDV and CPV, while 100% exhibited IgG against CDV when using the immunochromatographic technique. Regarding CAdV-1, 90% were seropositive for IgG, while 64% exhibited IgG against T. gondii. Nine dogs from the region were also sampled, and all (100%) exhibited IgM and IgG against CDV and CPV. For IgG against T. gondii and against CAdV-1, 90% of the animals were seropositive. Molecular evaluation yielded negative results for all maned wolves and dogs assessed for CAdV-1, CDV, and T. gondii, as well as the CCoV antigen. These data indicate the occurrence of viral agents and Toxoplasma gondii in maned wolves and dogs, suggesting circulation in both populations.

Detection and molecular analysis of Pseudorabies virus from free-ranging Italian wolves (Canis lupus italicus) in Italy - a case report

BackgroundThe only natural hosts of Pseudorabies virus (PRV) are members of the family Suidae (Sus scrofa scrofa). In mammals, the infection is usually fatal and typically causes serious neurologic disease. This study describes four Aujeszky’s disease cases in free-ranging Italian wolves (Canis lupus italicus). In Italy, the wolf is a strictly protected species and is in demographic expansion.Case presentationThree wolves (Wolf A, B, and C) were found in a regional park in Northern Italy, and one (Wolf D) was found in Central Italy. Wolf A and D were alive at the time of the finding and exhibited a fatal infection with epileptic seizures and dyspnoea, dying after a few hours. Wolf B presented scratching lesions under the chin and a detachment of the right earlobe, whilst Wolf C was partially eaten.The wolves showed hepatic congestion, diffuse enteritis, moderate pericardial effusion, severe bilateral pneumonia, and diffuse hyperaemia in the brain. The diagnostic examinations included virological analyses and detection of toxic molecules able to cause serious neurological signs.All four wolves tested positive for pseudorabies virus (PrV). The analysed sequences were placed in Italian clade 1, which is divided into two subclades, “a” and “b”. The sequences of Wolf A, B, and C were closely related to other Italian sequences in the subclade b, originally obtained from wild boars and hunting dogs. The sequence from Wolf D was located within the same clade and was closely related to the French hunting dog sequences belonging to group 4.ConclusionResults showed the presence of PrV strains currently circulating in wild boars and free-ranging Italian wolves. The genetic characterisation of the PrV UL44 sequences from the four wolves confirmed the close relationship with the sequences from wild boars and hunting dogs. This fact supports a possible epidemiological link with the high PrV presence in wild boars and the possibility of infection in wolves through consumption of infected wild boar carcasses or indirect transmission. To the best of our knowledge, this study is the first detection of Pseudorabies virus in free-ranging Italian wolves in northern and central Italy.

Tick-borne infections in wolves from an expanding population in Eastern Europe

In Central and Eastern Europe, wolf populations have been increasing over the last two decades, recolonizing areas from which the species had been previously exterminated. As wolves are still recovering after years of persecution by humans, recognizing pathogens infecting this species, including tick-borne infections, is crucial for its conservation. On the other hand the high mobility of wolves and their frequent contacts with humans, dogs, and other domestic species make them a potentially important zoonotic reservoir. In this paper, we used molecular methods to determine the prevalence of tick-borne pathogens in the following genera: Anaplasma, Babesia, Bartonella, Borrelia, and Rickettsia in 50 free-ranging wolves from Poland. We detected Babesia canis in the blood of nine individuals (prevalence 9/50=18 %). The obtained sequence showed the highest similarity to B. canis isolated from dogs and ticks, and all infected individuals originated from regions endemic to the ornate tick, Dermacentor reticulatus. Anaplasma phagocytophilum was found in tissue from one individual (1/50=2 %), and the sequence was assigned to the zoonotic ecotype I.

The Sarcoptic Mange in Maned Wolf (Chrysocyon brachyurus): Mapping an Emerging Disease in the Largest South American Canid.

The maned wolf (Chrysocyon brachyurus) is the largest South American canid. In Brazil, as in other countries, it is considered an endangered species. Habitat loss, landscape changes, hunting, and roadkill are the main threats to this species. In addition, invasive diseases of domestic animals are considered to be an emerging threat to the maned wolf, where parasitic diseases are relevant. Sarcoptic mange is a skin disease caused by the mite Sarcoptes scabiei. This disease is currently almost globally distributed, with a remarkable host diversity. In Brazil, reports of sarcoptic mange in wildlife include several species, both wild and captive. However, the impact of this disease on wildlife is unknown. At the time of writing, there is only one published report of sarcoptic mange in maned wolves. This study sheds light on the occurrence of sarcoptic mange in free-ranging maned wolves in their natural range. A total of 52 cases (suspected and confirmed) of sarcoptic mange were identified through social media review, camera trapping, chemical immobilization and sample collection. These cases were distributed in southeastern Brazil, in the states of São Paulo (n = 34), Minas Gerais (n = 17), and Rio de Janeiro (n = 1), demonstrating a rapid and widespread spread of this disease, although it still only occurs in part of the species' range. We expect that these results will help to subsidize future actions relevant to the control of this emerging disease.

The human-initiated model of wolf domestication - An expansion based on human-dingo relations in Aboriginal Australia.

The historically known relationship of interspecies companionship between Aboriginal foraging communities in Australia and free-ranging dingoes provides a model for understanding the human-canid relations that gave rise to the first domesticated dogs. Here, we propose that a broadly similar relationship might have developed early in time between wild-living wolves and mobile groups of foragers in Late Pleistocene Eurasia, with hunter-gatherers routinely raiding wild wolf dens for pre-weaned pups, which were socialized to humans and kept in camp as tamed companions ("pets"). We outline a model in which captive wolf pups that reverted to the wild to breed when they were sexually mature established their territories in the vicinity of foraging communities - in a "liminal" ecological zone between humans and truly wild-living wolves. Many (or most) of the wolf pups humans took from the wilderness to rear in camp may have derived from these liminal dens where the breeding pairs had been under indirect human selection for tameness over many generations. This highlights the importance of the large seasonal hunting/aggregation camps associated with mammoth kill-sites in Gravettian/Epigravettian central Europe. Large numbers of foragers gathered regularly at these locations during the wild wolf birthing season. We infer that if a pattern of this kind occurred over long periods of time then there might have been a pronounced effect on genetic variation in free-ranging wolves that denned and whelped in the liminal zones in the vicinity of these human seasonal aggregation sites. The argument is not that wolves were domesticated in central Europe. Rather, it is this pattern of hunter-gatherers who caught and reared wild wolf pups gathering seasonally in large numbers that might have been the catalyst for the early changes leading to the first domesticated dogs - whether in western Eurasia or further afield.

Key observations of flexed-leg urination in the free-ranging Gray Wolf (<i>Canis lupus</i>)

Flexed-leg urination (FLU) in female Gray Wolves (Canis lupus) has been little studied in the wild. Captive females in packs do not exhibit FLU unless they are both mature and dominant to an associate female, but these characteristics have not been confirmed in free-ranging wolves. We present observations of wolves in Yellowstone National Park that accord with those of wolves in captivity, extend our knowledge of FLU in Gray Wolf, pose additional questions about it, and suggest new areas of study to better understand it.

The origin and population genetics of wolves in the north Hungarian mountains

The grey wolf (Canis lupus) is one of the most challenging species to conserve in our modern and crowded world. Due to various factors, most European wolf populations are currently growing. In Hungary, numbers have increased since the 2000s. Although spontaneous recolonisation from Slovakia is considered to be the most likely mechanism by the majority of experts, some stakeholders claim that hand-reared individuals have been released. To determine the origin of wolves in northern Hungary, we analysed samples of free-ranging wolves collected in Slovakia and Hungary as well as samples from wolves in private enclosures in the region. We also included reference samples from domestic dogs. All samples were genotyped at 14 canine autosomal tetranucleotide microsatellite loci (STR) and analysed using multivariate, Bayesian methods. Hungarian wolf samples were also analysed using kinship methods. In the free-ranging wolf samples, all loci were polymorphic with 3–12 alleles. The overall observed (Ho) and unbiased expected (uHE) heterozygosities were 0.60–0.66 and 0.69–0.71, respectively. Parental and sibling relationships were also found among Hungarian individuals: three generations of a pack in the Bükk Mountains were identified. Samples from free-ranging wolves clustered separately from those of captive wolves and dogs. However, genetic similarities were found between Slovakian and Hungarian wolf samples. Our analyses indicate a Slovakian origin of the sampled Hungarian wolves, and we found no evidence that individuals originating in captivity have played any role in the recolonisation process. Kinship relationships and moderate genetic diversity suggest that there is ongoing gene flow across the Slovakian–Hungarian border.

Assessing Lead Exposure in Free-Ranging Gray Wolves (Canis lupus) in Minnesota, USA

The gray wolf (Canis lupus) is both an apex predator and a scavenger in Minnesota, US. Monitoring the health of Minnesota's gray wolf population is an important component of wolf management. Concern regarding whether wolves are being exposed to lead through scavenging viscera of hunter-harvested cervids left on the landscape, led to our study to determine lead-exposure rates. In fall 2012, livers from 147 hunter-harvested wolves (89 females, 58 males) were screened for lead and 20 other elements by inductively coupled plasma-atomic emission spectroscopy. Ten wolves (6.8%) were exposed to lead; only one had high enough exposure (6.14 ppm) to suggest lead toxicosis. Lead exposure varied by time of harvest, with nearly all lead-exposed wolves taken in the late hunting and trapping season (from 24 November 2012 to 31 January 2013), compared with the earlier hunting-only season (3-18 November 2012). Further, eight of 10 lead-exposed wolves were taken from deer-permit areas that harvested >1 deer/km2; only two of 10 were taken where deer harvest was less. This suggests the availability of viscera on the landscape may influence exposure risk of lead to wolves. More research is needed to determine baseline levels for toxic concentrations of lead in gray wolves and to determine clinical signs of lead poisoning in wild canids.

Integrated Use of Molecular Techniques to Detect and Genetically Characterise DNA Viruses in Italian Wolves (Canis lupus italicus).

Simple SummaryIn our study, different quantitative and qualitative molecular techniques were used to detect and genetically characterise Carnivore protoparvovirus 1, Canine adenovirus type 1 and 2 (CAdV-1 and CAdV-2), and Canine circovirus (CanineCV) in Italian wolves (Canis lupus italicus) of the Italian Apennines. Carnivore protoparvoviruses were the most frequently detected viruses, followed by CanineCV and CAdV. All the wolves tested positive for at least one of the DNA viruses screened, and 47.8% of the subjects were coinfected with two or three viruses. From viral sequences analysis, close correlations emerged between the viruses identified in the wolves and those circulating in domestic dogs, suggesting that the same viruses infect wolves and domestic dogs. Further studies are needed to investigate if pathogens are transmitted between the two species.In this study, internal organs (tongue, intestine, and spleen) of 23 free-ranging Italian wolves (Canis lupus italicus) found dead between 2017 and 2019 were tested for Carnivore protoparvovirus 1, Canine adenovirus (CAdV), and Canine circovirus (CanineCV) using real-time PCR assays. Genetic characterisation of the identified viruses was carried out by amplification, sequencing, and analysis of the complete viral genome or informative viral genes. All the wolves tested positive for at least one of the DNA viruses screened, and 11/23 were coinfected. Carnivore protoparvoviruses were the most frequently detected viruses (21/23), followed by CanineCV (11/23) and CAdV (4/23). From the analysis of the partial VP2 gene of 13 carnivore protoparvoviruses, 12 were canine parvovirus type 2b, closely related to the strains detected in dogs and wild carnivores from Italy, and one was a feline panleukopenia-like virus. Of the four CAdV identified, two were CAdV-1 and two were CAdV-2. The complete genome of seven CanineCVs was sequenced and related to the CanineCV identified in dogs, wolves, and foxes worldwide. Close correlations emerged between the viruses identified in wolves and those circulating in domestic dogs. Further studies are needed to investigate if these pathogens may be potentially cross-transmitted between the two species.

Field Anesthesia of the Maned Wolf (Chrysocyon brachyurus) in Bolivia.

Fifteen maned wolves (Chrysocyon brachyurus) were anesthetized a total of 43 times as part of a long-term ecology and health study in a remote region of northeastern Bolivia. We administered tiletamine-zolazepam (TZ) to wolves in box traps or free-ranging, from blinds or on foot, at a mean dosage of 4.6 mg/kg intramuscularly. Detailed anesthetic information was recorded in 24 of these events in 11 wolves (six males, five females), and wolves were monitored closely post procedure with very high frequency or global positioning system telemetry collars. Anesthetic induction was smooth and rapid in all cases, with a mean 6.4 min from injection to recumbency. Vital parameters were stable during the majority of procedures. As expected with this drug combination, recovery was long (mean time to standing 163 min [range: 80-235 min]) but smooth, and animals were monitored in most cases in box traps until stable for release. One case of apnea and prolonged recovery is reported. In two cases, wolves recovered normally but were found to move minimally in the 2.5-4 d postprocedure before resuming normal movements. Overall, TZ provided safe, stable immobilization of free-ranging maned wolves in remote and extreme field conditions, although postanesthesia monitoring via telemetry is recommended.

Wolf (Canis lupus) as canine adenovirus type 1 (CAdV-1) sentinel for the endangered cantabrian brown bear (Ursus arctos arctos).

Canine adenovirus type 1 (CAdV-1) causes infectious canine hepatitis (ICH) and has recently been described as a cause of death among endangered populations of European brown bear (Ursus arctos arctos) in the Cantabrian mountain range in Asturias, Spain. Sympatric wild and domestic carnivores can act as reservoirs of the virus and likely spread it into the environment and subsequently transmit it to brown bears. The present work investigates the prevalence and geo-temporal distribution of CAdV-1 among free-ranging wolves (Canis lupus) in Asturias from 2009 to 2018, during which three fatal cases of ICH were reported among brown bears in the region. A total of 149 wolves were analysed in this study, of which 21 (14%) were found to have CAdV-1 DNA based on real-time polymerase chain reaction (RT-PCR) of spleen samples. Prevalence of the virus was similar between males and females. All but one of the 20 CAdV-1-positive animals of estimable age were younger than 2years, and only one of the 46 adult animals (>2years) tested positive. Prevalence was highest in the western area of Asturias and during 2010 and 2011. Our results confirm that CAdV-1 is circulating in Asturian free-ranging wolves, supporting their possible role as virus reservoirs and sentinels in the region of this emerging disease in brown bears.

Molecular survey of tick-borne pathogens reveals a high prevalence and low genetic variability of Hepatozoon canis in free-ranging grey wolves (Canis lupus) in Germany

Wild carnivores are important hosts for ixodid ticks and presumed reservoirs for several tick-borne pathogens (TBPs) of medical and veterinary importance. However, little is known about the role that the European grey wolf (Canis lupus) plays in the enzootic cycle of TBPs. The recent recolonization of Central European lowland, including some regions in Germany, by the grey wolf, opened up an excellent opportunity for studying the impact of the wolf population on pathogen diversity and transmission dynamics. Therefore, in the present study, we investigated spleen samples collected from 276 grey wolves in 11 federal states in Germany for common TBPs by molecular methods. In total, 127 grey wolves (46.0 %) were found to be positive for Hepatozoon canis. Only two genetic variants of this protozoan parasite (herein designated as G1 and G2) were found to circulate among the grey wolves, which can be potentially shared between populations of domestic and other wild carnivores in the country. Two grey wolves were positive for Anaplasma phagocytophilum (0.7 %), and both were co-infected with H. canis G1 genotype. The presence of other pathogens could not be confirmed by PCR and sequencing. This study represents the first one reporting H. canis in a grey wolf population worldwide, and it provides highly relevant information, which may contribute to a better understanding of the epidemiology of TBPs and the pathogen transmission dynamics among the reintroduced population of grey wolves and other carnivores.

Characterising the harmonic vocal repertoire of the Indian wolf (Canis lupus pallipes).

Vocal communication in social animals plays a crucial role in mate choice, maintaining social structure, and foraging strategy. The Indian grey wolf, among the least studied subspecies, is a social carnivore that lives in groups called packs and has many types of vocal communication. In this study, we characterise harmonic vocalisation types of the Indian wolf using howl survey responses and opportunistic recordings from captive and nine packs (each pack contains 2–9 individuals) of free-ranging Indian wolves. Using principal component analysis, hierarchical clustering, and discriminant function analysis, we found four distinct vocalisations using 270 recorded vocalisations (Average Silhouette width Si = 0.598) which include howls and howl-barks (N = 238), whimper (N = 2), social squeak (N = 28), and whine (N = 2). Although having a smaller body size compared to other wolf subspecies, Indian wolf howls have an average mean fundamental frequency of 422 Hz (±126), which is similar to other wolf subspecies. The whimper showed the highest frequency modulation (37.296±4.601) and the highest mean fundamental frequency (1708±524 Hz) compared to other call types. Less information is available on the third vocalisation type, i.e. ‘Social squeak’ or ‘talking’ (Mean fundamental frequency = 461±83 Hz), which is highly variable (coefficient of frequency variation = 18.778±3.587). Lastly, we identified the whine, which had a mean fundamental frequency of 906Hz (±242) and is similar to the Italian wolf (979±109 Hz). Our study’s characterisation of the Indian wolf’s harmonic vocal repertoire provides a first step in understanding the function and contextual use of vocalisations in this social mammal.

Seasonal Cycles in Hematology and Body Mass in Free-Ranging Gray Wolves (Canis lupus) from Northeastern Minnesota, USA

Studies of captive gray wolves (Canis lupus) showed seasonal cycles in hematologic values and female body mass. We used a remotely controlled recapture collar to determine whether nine female and five male free-ranging wolves handled four to 17 times in NE Minnesota, US showed similar cycles. Hematocrit, hemoglobin, red blood cell count, mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration, and body mass increased from summer toward a winter peak and then decreased again toward summer. Several hematologic values differed considerably from those of captive wolves, and the ranges in free-ranging wolves were much greater than those of captives.

GPS-coupled contaminant monitors on free-ranging Chernobyl wolves challenge a fundamental assumption in exposure assessments

Measurements of external contaminant exposures on individual wildlife are rare because of difficulties in using contaminant monitors on free-ranging animals. Most wildlife contaminant exposure data are therefore simulated with computer models. Rarely are empirical exposure data available to verify model simulations, or to test fundamental assumptions inherent in exposure assessments. We used GPS-coupled contaminant monitors to quantify external exposures to individual wolves (Canis lupus) living within the Belarus portion of Chernobyl's 30-km exclusion zone. The study provided data on animal location and contaminant exposure every 35 min for 6 months, resulting in ~6600 individual locations and 137Cs external exposure readings per wolf, representing the most robust external exposure data published to date on free ranging animals. The data provided information on variation in external exposure for each animal over time, as well as variation in external exposure among the eight wolves across the landscape of Chernobyl. The exposure data were then used to test a fundamental assumption in screening-level risk assessments, espoused in guidance documents of the U.S. Environmental Protection Agency and U.S. Department of Energy, — Mean contaminant concentrations conservatively estimate individual external exposures. We tested this assumption by comparing our empirical data to a series of simulations using the ERICA modeling tool. We found that modeled simulations of mean external exposure (10.5 mGy y−1), based on various measures of central tendency, under-predicted mean exposures measured on five of the eight wolves wearing GPS-contaminant monitors (i.e., 12.3, 26.3, 28.0, 28.8 and 35.7 mGy y−1). If under-prediction of exposure occurs for some animals, then arguably the use of averaged contaminant concentrations to predict external exposure is not as conservative as proposed by current risk assessment guidance. Thus, a risk assessor's interpretation of simulated exposures in a screening-level risk analysis might be misguided if contaminant concentrations are based on measures of central tendency. We offer three suggestions for risk assessors to consider in order to reduce the probability of underestimating exposure in a screening-level risk assessment.

Helminth Fauna in Captive European Gray Wolves (Canis lupus lupus) in Germany.

Captive as well as free-ranging wolves, which are currently recolonizing Germany, may harbor a variety of gastrointestinal parasites. This study investigated endoparasites in captive European gray wolves (Canis lupus lupus) using coproscopical methods. Fecal samples were collected monthly between October 2012 and November 2013 from 18 wolf enclosures in 14 German zoological gardens, representing 72 individual wolves. In total, 1,041 fecal samples including 26 bulk samples were analyzed by the sedimentation and flotation method. The most frequently detected egg morphotypes included five nematodes [Ancylostomatidae (Ancylostoma or Uncinaria spp.), Toxocara canis, Toxascaris leonina, Trichuris vulpis, and Capillaria/Eucoleus spp.], one cestode (Taeniidae) and one trematode (Alaria alata). 44.76% of all samples were positive for at least one of these egg morphotypes. Overall, Ancylostomatidae showed the highest frequency (30.84% of all samples), followed by Capillaria/Eucoleus spp. (19.88%), Toxocara canis (5.19%), taeniids (3.75%), Trichuris vulpis and Alaria alata (3.65% each), and Toxascaris leonina (1.25%). As fecal samples were collected from the environment and could not be assigned to individual wolves, sample results were combined per zoo and month. General linear mixed models were employed to analyze the effect of season and management factors on the occurrence of Ancylostomatidae, Capillaria/Eucoleus spp., Toxocara canis and taeniids. No statistically significant effect of season was found, whereas anthelmintic treatment negatively affected Ancylostomatidae egg excretion. Detected parasites and their prevalences are comparable to previous studies on wolf parasitism conducted elsewhere in Europe. As many of the most prevalent helminths are of zoonotic importance, routine anthelmintic treatment of captive wolves should be recommended.

Conservation implications for the Himalayan wolf Canis (lupus) himalayensis based on observations of packs and home sites in Nepal

AbstractWe provide insights into pack composition and den site parameters of the Himalayan wolf Canis (lupus) himalayensis based on observations of free-ranging wolves in three study areas in Nepal. We combine this with a social survey of the local Buddhist communities regarding human–carnivore conflict, to draw inferences for conservation practice in the Nepalese Himalayas. We recorded eight wolf packs (with an average composition of two adults and three pups), and found five home sites in high-altitude shrubland patches within alpine grasslands at 4,270–4,940 m altitude. There was a spatial–temporal overlap of wolf home sites and livestock herding during spring and summer, which facilitated human–wolf conflict. The litters of three out of five wolf packs found in Dolpa during 2016 were killed by local people in the same year. In Nepal compensation is offered for depredation by snow leopards Panthera uncia, with associated lowering of negative attitudes, but not for depredation by wolves. We recommend the implementation of financial and educational conservation schemes for all conflict-causing carnivores across the Himalayan regions of Nepal.

Viral gut metagenomics of sympatric wild and domestic canids, and monitoring of viruses: Insights from an endangered wolf population.

Animal host–microbe interactions are a relevant concern for wildlife conservation, particularly regarding generalist pathogens, where domestic host species can play a role in the transmission of infectious agents, such as viruses, to wild animals. Knowledge on viral circulation in wild host species is still scarce and can be improved by the recent advent of modern molecular approaches. We aimed to characterize the fecal virome and identify viruses of potential conservation relevance of diarrheic free‐ranging wolves and sympatric domestic dogs from Central Portugal, where a small and threatened wolf population persists in a highly anthropogenically modified landscape. Using viral metagenomics, we screened diarrheic stools collected from wolves (n = 8), feral dogs (n = 4), and pet dogs (n = 6), all collected within wolf range. We detected novel highly divergent viruses as well as known viral pathogens with established effects on population dynamics, including canine distemper virus, a novel bocavirus, and canine minute virus. Furthermore, we performed a 4‐year survey for the six wolf packs comprising this endangered wolf population, screening 93 fecal samples from 36 genetically identified wolves for canine distemper virus and the novel bocavirus, previously identified using our metagenomics approach. Our novel approach using metagenomics for viral screening in noninvasive samples of wolves and dogs has profound implications on the knowledge of both virology and wildlife diseases, establishing a complementary tool to traditional screening methods for the conservation of threatened species.

The prevalence and impact of Babesia canis and Theileria sp. in free-ranging grey wolf (Canis lupus) populations in Croatia

BackgroundBabesia spp. and Theileria spp. are important emerging causes of disease in dogs. Alongside these domesticated hosts, there is increasing recognition that these piroplasms can also be found in a range of wild animals with isolated reports describing the presence of these pathogen in foxes (Vulpes vulpes) and captive grey wolves (Canis lupus). The prevalence and impact of these infections in free-ranging populations of canids are unknown. To gain a better insight into the epidemiology and pathogenesis of piroplasm infections in free-ranging grey wolves, pathological and molecular investigations into captive and free-ranging grey wolves in Croatia were performed.ResultsThe carcasses of 107 free-ranging wolves and one captive wolf were the subjects of post-mortem investigations and sampling for molecular studies. A blood sample from one live captured wolf for telemetric tracking was also used for molecular analysis. PCR amplification targeting the 18S RNA gene revealed that 21 of 108 free-ranging wolves and one captive animal were positive for Theileria/Babesia DNA. Subsequent sequencing of a fragment of the 18S RNA gene revealed that 7/22 animals were positive for Babesia canis while the other amplified sequence were found to be identical with corresponding 18S rDNA sequences of Theileria capreoli isolated from wild deer (15/22). Haematological and cytological analysis revealed the presence of signet-ring shaped or pear-shaped piroplasms in several animals with the overall parasite burden in all positive animals assessed to be very low. Pathological investigation of the captive animal revealed fatal septicemia as a likely outcome of hemolytic anaemia. There was little or no evidence of hemolytic disease consistent with babesiosis in other animals.ConclusionImportantly, the presence of B. canis in free-ranging grey wolves has not been described before but has been reported in a single fox and domestic dogs only. That B. canis infections cause disease in dogs but have little impact on wolf health possibly suggests that the wolf is the natural and the domestic dog is a secondary host. Surprisingly, the frequent finding of Theileria capreoli in wolves suggests that this Theileria species is not restricted to ungulates (cervids) but commonly infects also this carnivore species. Nevertheless, the potential role that these asymptomatically infected animals may play in the dispersal of these pathogens to susceptible sympatric species such as domesticated dogs requires further investigation.