Molecular Characterization of Enterocytozoon bieneusi in Wild Carnivores in Spain

Microsporidia comprises a diverse group of obligate intracellular parasites that infect a broad range of invertebrates and vertebrates. Among Microsporidia, Enterocytozoon bieneusi is the most frequently detected species in humans and animals worldwide bringing into question the possible role of animal reservoirs in the epidemiology of this pathogen. Although E. bieneusi is an emerging zoonotic pathogen able to infect many domestic and wild mammals that could act as reservoir of infection for humans and other animals, only few studies have documented its occurrence in wild carnivores. To determine the occurrence of E. bieneusi in wild carnivores, we examined 190 wild carnivores collected from different locations in Spain. Twenty‐five fecal samples (13.2%) from three host species (European badger, beech marten, and red fox) were E. bieneusi‐positive by PCR. Nucleotide sequence analysis of the ITS region revealed a high degree of genetic diversity with a total of eight distinct genotypes including four known (PtEbIX, S5, S9, and WildBoar3) and four novel (EbCar1‐EbCar4) genotypes identified. Phylogenetic analysis showed that the four novel genotypes (EbCar1‐EbCar4), S5, S9, and WildBoar3 clustered within the previously designated zoonotic Group 1. Our results demonstrate that human‐pathogenic genotypes are present in wild carnivores, corroborating their potential role as a source of human infection and environmental contamination.     

Exposure of free-ranging wild carnivores, horses and domestic dogs to Leptospira spp in the northern Pantanal, Brazil

Leptospirosis is a zoonotic disease affecting most mammals and is distributed throughout the world. Several species of domestic and wild animals may act as reservoirs for this disease. The purpose of this study was to assess the exposure of free-ranging wild carnivores, horses and domestic dogs on a private reserve located in the northern Pantanal (Brazil) and the surrounding areas to Leptospira spp from 2002-2006, 75 free-ranging wild carnivores were captured in the Pantanal and serum samples were collected. In addition, samples from 103 domestic dogs and 23 horses in the region were collected. Serum samples were tested for the presence of Leptospira antibodies using the microscopic agglutination test. Thirty-two wild carnivores (42.7%) were considered positive with titres ≥ 100, and 18 domestic dogs (17.5%) and 20 horses (74.1%) were also found to be positive. Our study showed that horses, dogs and several species of free-ranging wild carnivores have been exposed to Leptospira spp in the Pantanal, suggesting that the peculiar characteristics of this biome, such as high temperatures and an extended period of flooding, may favour bacterial persistence and transmission. In this region, wild carnivores and horses seem to be important hosts for the epidemiology of Leptospira species.     

Detection of rabies virus antibodies in Brazilian free-ranging wild carnivores

Rabies virus is a pathogen of major concern in free-ranging wild carnivores in several regions of the world, but little is known about its circulation in Brazilian wild carnivores. Sera from 211 free-ranging wild carnivores, captured from 2000 to 2006 in four locations of two Brazilian biomes (Pantanal and Cerrado), were tested for rabies antibodies. Twenty-six individuals (12.3%) had neutralizing antibody titers ≥0.10 IU/ml. The four sampled locations had antibody-positive animals, suggesting that Rabies virus circulates in all of these regions. Results underscore the risk posed by rabies for conservation of Brazilian carnivores and the possibility of the animals acting as reservoirs for the Rabies virus.     

Sarcoptic mange vulnerability in carnivores of the Białowieża Primeval Forest,Poland: underlying determinant factors

Sarcoptic mange caused by Sarcoptes scabiei, is a highly contagious worldwide mite infection responsible for epizootic skin disease in populations of wild and domestic mammals. It causes skin disorders that may lead to severe generalized skin disease (mange). We investigated the spread and dynamics of sarcoptic mange in one of the best preserved carnivore communities in the Bia?owie?a Primeval Forest over the last 20 years. Sarcoptic mange was found in 6 of 10 investigated species. The highest percentage of infected individuals was recorded in red foxes (19 %), European badgers (9 %), grey wolves (7 %), raccoon dogs (6 %), and stone martens (5 %). Other species were not infected (least weasel, American mink, European polecat, pine marten) or infected sporadically (Eurasian lynx––1 case). Rate of infection was correlated with the sum of ecological and behavioral features, which may increase the spread of infection. Outbreaks of sarcoptic mange occurred simultaneously in several species of carnivores, which indicates the occurrence of between species transmission. We found that the source of investigated individuals (trapped, culled and found dead) may influence the results of the analysis. Our study shows that in carnivore communities such as in Bia?owie?a Forest, numerous species may serve as a reservoir of the pathogen and favor its persistence in the environment. This study indicates that sociality and use of den sites are important factors affecting sarcoptic mange spread and prevalence in wild populations of carnivores.     

Absence of antibodies against canine distemper virus in free-ranging populations of the Eurasian badger in Great Britain

Canine distemper virus (CDV) is a serious disease of wild carnivores throughout the world. In Europe, infection has been detected in several carnivores including the Eurasian badger (Meles meles). In the present study 182 badger blood samples were collected from an intensively studied population of wild badgers in southwestern England (January-July, 1997), and a further 286 from throughout southern Britain (June 1996-November 1998). A neutralizing peroxidase-linked antibody test was used for the detection of antibodies against CDV. All the samples were negative for CDV antibodies, suggesting that in contrast to mainland Europe, the disease may be either absent or maintained at low levels in British badgers.     

Prey preference of large carnivores in Anamalai Tiger Reserve,India

Prey preferences of large carnivores (tiger (Panthera tigris), leopard (Panthera pardus) and dhole (Cuon alpinus)) in the tropical forest of Anamalai Tiger Reserve (ATR) were evaluated. This was the first study in ATR to estimate the density of prey and the food habits of these large carnivores. The 958-km² intensive study area was found to have a high mammalian prey density (72.1 animals per square kilometre) with wild boar (20.61 animals per square kilometre) and chital (20.54 animals per square kilometre) being the most common species, followed by nilgiri tahr (13.6 animals per square kilometre). When the density figures were multiplied by the average weight of each prey species, a high biomass density of 14,204 kg km⁻² was obtained for the intensive study area. Scat analysis and incidental kill observation were used to determine the dietary composition of these predators. During the study from the period of March 2001 to April 2004, 1,145 tiger scats, 595 leopard scats and 2,074 dhole scats were collected and analysed. Kill data were based on direct observation of 66 tiger kills and 39 leopard kills. Sambar, with a density of 6.54 kg km⁻² was the preferred prey for these carnivores. Sambar constitutes 35% of the overall diet of tiger, whereas it constitutes 17% and 25% in leopard and dhole diets, respectively. Chital was utilized less than sambar in the range of about 7%, 11% and 15% by tiger, leopard and dhole, respectively. Predator diet was estimated more accurately by scat analysis, which reveals 30% of smaller prey species in leopard’s diet, which was not observed by kill data. This study reveals that ATR harbours high prey density, and these large carnivores seem mostly dependent on the wild prey rather than on domestic livestock as in some other areas in the subcontinent. These factors make ATR a potential area for long-term conservation of these endangered carnivores.     

First report of Thelazia callipaeda (Spirurida, Thelaziidae) in wolves in Italy

Thelazia callipaeda (Spirurida, Thelaziidae) infects the eyes of humans and rabbits as well of domestic and wild carnivores (i.e., dogs, cats, and foxes). The first three cases of infection by T. callipaeda eyeworms in wolves (Canis lupus) are here described with up to 96 nematodes collected from a single animal. The host competence of wolf was demonstrated by the retrieval of adult worms in the eyes of all examined animals. All nematodes collected were evaluated by morphologic and molecular techniques. Sequence analysis of the partial mitochondrial cytochrome c oxidase subunit 1 gene (cox1) confirmed that only one haplotype of T. callipaeda was present. The competence of wolves as a definitive host for T. callipaeda is discussed in view of the relevance of wild fauna in maintaining and spreading eyeworm infection in humans and domestic animals.     

Epizootiologic investigations of parvovirus infections in free-ranging carnivores from Germany

To assess if wild carnivores in Germany play a role in the epizootiology of canine parvovirus (CPV) infection, seroprevalences against CPV in free-ranging carnivores (n=1,496) from selected urban and rural areas were compared. Antibodies against CPV were found in sera from red foxes (Vulpes vulpes; 136 of 1,442; 9%), raccoon dogs (Nyctereutes procyonides; two of 33; 6%), stone martens (Martes foina; four of 13; 31%), and pine martens (Martes martes; one of two) using the hemagglutination-inhibition test and pig erythrocytes. Evidence of CPV infection was detected in all study areas. Antibody titers varied between 10 and 320. In red foxes, the number of reactors did not differ between most urban and rural areas. However, we found significantly more reactors in the most densely populated urban area (Berlin). None of 430 tissue samples (small intestine, spleen, mesenterial lymph nodes) from any species tested for the presence of CPV nucleic acid using polymerase chain reaction yielded an amplification product. Based on our results, we believe that contact between domestic dogs and free-ranging red foxes probably plays a subordinate role in the epizootiology of CPV in Germany.     

Alveolar echinococcosis in Belorussian polesie

Results of long-time (1980-1998) investigations of wild animals, which are potential hosts of the pathogen of alveolar echinococcosis in Belorussian Polesie, have been already generalized. One species of carnivores, two species of insectivores and eight species of rodents take part in circulation of infection.     

Using Molecular Epidemiology to Track Toxoplasma gondii from Terrestrial Carnivores to Marine Hosts: Implications for Public Health and Conservation

Background

Environmental transmission of the zoonotic parasite Toxoplasma gondii, which is shed only by felids, poses risks to human and animal health in temperate and tropical ecosystems. Atypical T. gondii genotypes have been linked to severe disease in people and the threatened population of California sea otters. To investigate land-to-sea parasite transmission, we screened 373 carnivores (feral domestic cats, mountain lions, bobcats, foxes, and coyotes) for T. gondii infection and examined the distribution of genotypes in 85 infected animals sampled near the sea otter range.

Methodology/Principal Findings

Nested PCR-RFLP analyses and direct DNA sequencing at six independent polymorphic genetic loci (B1, SAG1, SAG3, GRA6, L358, and Apico) were used to characterize T. gondii strains in infected animals. Strains consistent with Type X, a novel genotype previously identified in over 70% of infected sea otters and four terrestrial wild carnivores along the California coast, were detected in all sampled species, including domestic cats. However, odds of Type X infection were 14 times higher (95% CI: 1.3–148.6) for wild felids than feral domestic cats. Type X infection was also linked to undeveloped lands (OR = 22, 95% CI: 2.3–250.7). A spatial cluster of terrestrial Type II infection (P = 0.04) was identified in developed lands bordering an area of increased risk for sea otter Type II infection. Two spatial clusters of animals infected with strains consistent with Type X (P≤0.01) were detected in less developed landscapes.

Conclusions

Differences in T. gondii genotype prevalence among domestic and wild felids, as well as the spatial distribution of genotypes, suggest co-existing domestic and wild T. gondii transmission cycles that likely overlap at the interface of developed and undeveloped lands. Anthropogenic development driving contact between these cycles may increase atypical T. gondii genotypes in domestic cats and facilitate transmission of potentially more pathogenic genotypes to humans, domestic animals, and wildlife.     

Ethiopian wolves conflict with pastoralists in small Afroalpine relicts

The peaceful coexistence between people and the rare Ethiopian wolf Canis simensis is being challenged by conflicts rising due to livestock predation by wild carnivores. Understanding the cultural and socio‐economic context of these conflicts can help to prevent negative attitudes and retaliatory killings, which have the potential to seriously compromise the survival of Ethiopian wolf populations in small Afroalpine relicts. With this objective in mind, questionnaire surveys were conducted in 140 households around the Aboi Gara range in north Ethiopia. Half of the households reported losing sheep and goats to wolves and golden jackals Canis aureus, with an annual average loss of 1.2 heads per year (10% of the average herd size), equivalent to 92 US$. Aboi Gara pastoralists considered wolves and jackals to be equally responsible for livestock killings. Households with large herds, closer to Afroalpine habitats, and using Afroalpine pastures for longer periods, reported more predation by wild carnivores. Most respondents (62%) expressed a positive attitude towards Ethiopian wolves, particularly literate people and those with smaller herds. We suggest ways to diminish conflict, including best livestock guarding techniques to lessen the risk of livestock predation by wild carnivores in Afroalpine areas.     

Exposure of Free-Ranging Wild Carnivores and Domestic Dogs to Canine Distemper Virus and Parvovirus in the Cerrado of Central Brazil

Human population growth around protected areas increases the contact between wild and domestic animals, promoting disease transmission between them. This study investigates the exposure of free-ranging wild carnivores and domestic dogs to canine distemper virus (CDV) and parvovirus in Emas National Park (ENP) in the Cerrado savanna of central Brazil. Serum samples were collected from 169 wild carnivores, including the maned wolf (Chrysocyon brachyurus), crab-eating fox (Cerdocyon thous), hoary fox (Pseudalopex vetulus), puma (Puma concolor), ocelot (Leopardus pardalis), pampas cat (Leopardus colocolo), jaguarundi (Herpailurus yagouaroundi), striped hog-nosed skunk (Conepatus semistriatus) and coati (Nasua nasua), and from 35 domestic dogs living on rural properties bordering ENP. Serological tests showed that 10.6% of wild carnivores (maned wolves, crab-eating foxes and ocelots) and 71.4% of domestic dogs were exposed to CDV, and 56.8% of wild carnivores, including all species sampled except coatis, and 57.1% of domestic dogs were exposed to parvovirus. This report is the first to indicate that the free-ranging pampas cat, jaguarundi and striped hog-nosed skunk are exposed to parvovirus. CDV and parvovirus deserve attention in ENP, and it is extremely important to monitor the health of carnivore populations and perform molecular diagnosis of the viruses to determine the possible involvement of the domestic dog in their transmission.     

Prevalence and Phylogenetic Analysis of <Emphasis Type="Italic">Bartonella</Emphasis> Species of Wild Carnivores and Their Fleas in Northwestern Mexico

The host–parasite–vector relationship of Bartonella spp. system in wild carnivores and their fleas from northwestern Mexico was investigated. Sixty-six carnivores belonging to eight species were sampled, and 285 fleas belonging to three species were collected during spring (April–May) and fall (October–November) seasons. We detected Bartonella species in 7 carnivores (10.6%) and 27 fleas (9.5%) through either blood culture or PCR. Of the 27 Bartonella-positive fleas, twenty-two were Pulex simulans, three were Pulex irritans and one was Echidnophaga gallinacea. The gltA gene and ITS region sequences alignment revealed six and eight genetic variants of Bartonella spp., respectively. These variants were clustered into Bartonella rochalimae, Bartonella vinsonii subsp. berkhoffii and another genotype, which likely represents a novel species of Bartonella spp. Although experimental infection studies are required to prove the vector role of P. simulans, our results suggest that this flea may play an important role in the Bartonella transmission. The results indicated possible host-specific relationships between Bartonella genotypes and the families of the carnivores, but further studies are needed to verify this finding. The presence of zoonotic species of Bartonella spp. in wild carnivores raises the issue of their potential risk for humans in fragmented ecosystems.     

Livestock predation in the Bale Mountains,Ethiopia

In the Web Valley of the Bale Mountains National Park, the pastoral people suffer from livestock predation by wild carnivores. A total of 704 livestock were reported to be killed by wild carnivores over a 3‐year period, causing a loss of potential revenue of 12 USD per year per household. Reported annual predation rates equated to 1.4% of the livestock population of the study area. Spotted hyaenas were responsible for most livestock predation (57%), followed by leopards (18%), common jackals (16%) and servals (9%). Hyaenas killed all livestock types (horses, donkeys, mules, cattle, goats and sheep) whilst leopards, common jackals and servals killed mostly goats and sheep. A survey of 362 households revealed that the pastoral people keep dogs to protect livestock from carnivores. During 250 nights of observation in the ten settlements, pastoralists were alerted to the presence of hyaenas on 80 occasions by the barking of their dogs. Such tradition of keeping dogs presents a threat to the persistence of the endangered Ethiopian wolf through diseases transmission. Given the frequency of carnivore attacks on livestock, it is desirable to develop alternative livestock protection methods that both minimize livestock losses and reduce the risk of disease transmission to Ethiopian wolves.     

Trophic niche overlap and wild ungulate consumption by red fox and wolf in a mountain area in Italy

Red fox (Vulpes vulpes) and wolf (Canis lupus) are two widespread opportunistic predators living in simpatry in many areas. Nonetheless, scarce information are available on their trophic interactions. We investigated food habits of these two carnivores in a mountain area in Italy and assessed the extent of their trophic niche overlap, focusing on the consumption of wild ungulates. Thereby we analyzed the content of 669 red fox scats and 253 wolf scats collected between May 2008 and April 2009. Red foxes resulted to have a more than three times higher niche breadth than wolves. Vegetables, small mammals, wild ungulates, and invertebrates were major items (altogether 92% of volume) of the red fox annual diet. On the contrary wolf annual diet relied on wild ungulates (94% of volume) with wild boar (Sus scrofa) being the main food item. The degree of trophic niche overlap between the two species was found to be low (Pianka's O = 0.356). Diet variation between the warm and the cold seasons was limited in both species, and higher in red fox than in wolf. The two canids appeared to use wild ungulates unevenly being the former more selective for younger preys, smaller in size (newborn piglets and roe deer Capreolus capreolus fawns), whereas the latter exhibited a preference for medium-sized and large ungulates (10–35 kg wild boar and adult roe deer). Even if wild ungulates represent the main shared food category, the different use of age/weight classes by the two predators, together with their possible consumption as carrions by red fox, suggests a very limited trophic competition between wolf and red fox.This study represents a contribution to the knowledge of trophic interaction in predator–prey systems where sympatric carnivores are present.     

Antibodies to Toxoplasma gondii and Neospora caninum in captive neotropical and exotic wild canids and felids

This study was designed to detect antibodies to Toxoplasma gondii and Neospora caninum in wild captive carnivores maintained in Brazilian zoos. Blood samples were collected from 142 Brazilian wild felids and 19 exotic felids in zoos, and 3 European wolves (Canis lupus) and 94 Brazilian wild canids maintained in captivity in Brazilian zoos of S?o Paulo, Mato Grosso states and Federal District. One hundred and two (63.4%) and 70 (50.3%) of the 161 wild felids tested were seropositive for T. gondii and N. caninum by indirect immunofluorescent assay test (IFAT), respectively. Among sampled wild canids, 49 (50.5%) and 40 (41.2%) animals were seropositive for T. gondii and N. caninum antigens by IFAT, respectively. To our knowledge, this is the first serological detection of antibodies to N. caninum in Brazilian wild captive felids and bush dogs (Speothos venaticus (Lund)).     

Trichinosis in South-west Ireland

Since the large outbreak of trichinosis in Liverpool in 1953 there have been three outbreaks in Ireland, two occurring recently in Cork and Kerry, where a total of 76 cases was confirmed. In the outbreak of 50 cases in Kerry the latex trichina slide agglutination test was found to be valuable diagnostically and epidemiologically, while the efficacy of thiabendazole treatment was not established. This infection may be maintained as a reservoir in wild carnivores, occasionally spreading to pigs and perpetuated in them by a failure to boil pig swill.     

Sarcocystis and Its Complications in Camels (Camelus dromedarius) of Eastern Provinces of Iran

The prevalence of Sarcocystis spp. was investigated by gross and histopathological examinations in 250 camels (Camelus dromedarius) slaughtered from 2002 to 2005 in the Mashhad Slaughterhouse, eastern Iran. Samples were taken from the diaphragm, heart, tongue, esophagus and masseter muscles for histopathological studies. No macroscopic sarcocysts were found in the samples at gross inspection. Sarcocysts were detected in 209 of 250 (83.6%) examined camels at histopathological level. The infection rate of the esophagus, heart, masseter muscles, diaphragm, and tongue was 58.8%, 48.0%, 46.8%, 41.6%, and 28.0%, respectively. There was no significant difference in the rate of infection between male (85.8%) and female (81.0%) camels. The tissue response to vital cysts was minimal; however, reaction to the degenerating cysts was severe and caused tissue damages resulting in hyperemia, hemorrhages, mononuclear cell infiltration, necrotic changes, and fibrosis. The wild and domestic carnivores especially dogs may be the final hosts of Sarcocystis spp. in this area.     

Presence of Bartonella species in wild carnivores of northern Spain

The genus Bartonella was detected by PCR in 5.7% (12/212) of wild carnivores from Northern Spain. Based on hybridization and sequence analyses, Bartonella henselae was identified in a wildcat (Felis silvestris), Bartonella rochalimae in a red fox (Vulpes vulpes) and in a wolf (Canis lupus), and Bartonella sp. in badgers (Meles meles).     

Frugivory and spatial patterns of seed deposition by carnivorous mammals in anthropogenic landscapes: a multi-scale approach

Background

Knowledge about how frugivory and seed deposition are spatially distributed is valuable to understand the role of dispersers on the structure and dynamics of plant populations. This may be particularly important within anthropogenic areas, where either the patchy distribution of wild plants or the presence of cultivated fleshy-fruits may influence plant-disperser interactions.

Methodology/Principal Findings

We investigated frugivory and spatial patterns of seed deposition by carnivorous mammals in anthropogenic landscapes considering two spatial scales: ‘landscape’ (∼10 km²) and ‘habitat type’ (∼1–2 km²). We sampled carnivore faeces and plant abundance at three contrasting habitats (chestnut woods, mosaics and scrublands), each replicated within three different landscapes. Sixty-five percent of faeces collected (n = 1077) contained seeds, among which wild and cultivated seeds appeared in similar proportions (58% and 53%) despite that cultivated fruiting plants were much less abundant. Seed deposition was spatially structured among both spatial scales being different between fruit types. Whereas the most important source of spatial variation in deposition of wild seeds was the landscape scale, it was the habitat scale for cultivated seeds. At the habitat scale, seeds of wild species were mostly deposited within mosaics while seeds of cultivated species were within chestnut woods and scrublands. Spatial concordance between seed deposition and plant abundance was found only for wild species.

Conclusions/Significance

Spatial patterns of seed deposition by carnivores differed between fruit types and seemed to be modulated by the fleshy-fruited plant assemblages and the behaviour of dispersers. Our results suggest that a strong preference for cultivated fruits by carnivores may influence their spatial foraging behaviour and lower their dispersal services to wild species. However, the high amount of seeds removed within and between habitats suggests that carnivores must play an important role – often overlooked – as ‘restorers’ and ‘habitat shapers’ in anthropogenic areas.