Canine distemper virus may affect European wild cat populations in Central Spain.

The main objective of this brief communication is to inform about the exposure to certain pathogens of interest for mesocarnivores in wildcats (Felis silvestris silvestris) that inhabit a human-domestic-wild ecotone located in a Natural Park (Serranía de Cuenca, Central Spain). Blood and mucosal swabs (nasal, conjunctival and rectal) samples were collected from nine alive animals to detect canine distemper virus (CDV), parvovirus (CPV/FPV), feline leukaemia virus (FeLV), feline coronavirus (FCoV), feline immunodeficiency virus (FIV), Leptospira interrogans, Chlamydia felis, Ehrlichia canis, Toxoplasma gondii, and Neospora caninum. ELISA, immunochromatograpy, microscopy agglutination test and PCR assays were used. The results show the first worldwide detection of exposure of wildcats to L. interrogans (3 positive/9 analysed) and the first detection of exposure to CVD (7/9), of carriers ofC felis (2/9) and of fecal spreading of CPV-FPV (2/9) in wildcats in Spain. Exposure to T. gondii and CPV-FPV was detected in 5 of the 9 wildcats analysed, and to FelV in 4 of 9. No FIV, FCoV, Ehrlichia canis and Neospora caninum were detected. The results reveal the circulation of pathogens among the wildcat population studied, but more vigilance is needed for an accurate assessment of the impact of these pathogens on the health status of this population.

Using a top predator as a sentinel for environmental contamination with pathogenic bacteria: the Iberian wolf and leptospires.

The Iberian wolf (Canis lupus) is the top predator in the Iberian environments in which it lives, feeding on a wide range of species, thus encountering a wide range of disease agents. Therefore, the wolf can serve as sentinel of environmental contamination with pathogens. We investigated the exposure of free-living wolves to 14 serovars of Leptospira interrogans sensu lato. Kidney samples from 49 wolves collected from 2010-2013 in northwestern Spain were analysed by culture, direct immunofluorescence and polymerase chain reaction. Tissue fluids were analysed for antibodies by a microscopic agglutination test. Ten wolves (observed prevalence: 20%, 95% confidence interval = 11-33%) showed evidence of contact with leptospires, eight through direct detection and nine through serology (7 wolves were positive according to both techniques). Titres below the cut-off level were also detected in seven cases. Serovars confirmed were Canicola (n = 4), Icterohaemorrhagiae (n = 3) and Sejroë, Ballum and Grippotyphosa (n = 1 each), indicating that wolves were infected with serovars for which dogs, rodents and ungulates, are the natural hosts and supporting the utility of the wolf and other large predators as environmental sentinels for pathogens.

Urban landscape and infection risk in free-roaming cats.

Despite public concern on the role of free-roaming cats as reservoirs of zoonotic agents, little is known about the influence of urban and peri-urban landscapes on the exposure risk. We evaluated the seroprevalence of three zoonotic agents (Chlamydia felis, Coxiella burnetii and Toxoplasma gondii) in domestic cats (Felis catus). Two hundred and ninety-one free-roaming cats were trapped in Murcia municipality (Southeast Spain), and their sera were tested for specific antibodies against T. gondii using a modified agglutination test (MAT), and for C. felis, C. burnetii and feline immunodeficiency virus (FIV) antibodies with ELISA technique. Pathogen seroprevalence at 95% CI was calculated for each sex and age category (up to and over 12 months) and compared with a chi-squared test. The role of human population density and urban landscape characteristics on the risk of pathogen exposure in the cat population was explored using generalized linear models. Seropositivity against a single pathogen was found in 60% of the cats, while 19% was seropositive for two or three pathogens. Seroprevalence of C. felis was 8% (CI95% : 5-11), 37% (CI95% : 31-42) for C. burnetii and 42% (CI95% : 36-47) for T. gondii. In addition to these three pathogens, FIV seropositivity was low (1%, CI95% : -0.1 to 2) and adult cats were more likely to be seropositive to C. burnetii than young individuals (OR: 2.3, CI95% : 1.2-4.2). No sex or age class differences in seroprevalence were observed for the rest of the pathogens. Seropositivity was correlated with water surface areas for C. felis, and not with crop areas. Coxiella burnetii seropositivity was correlated with the percentage of urban areas (continuous with only buildings and discontinuous, that include buildings, parks, and pedestrian and urban green areas), human population size and peri-urban areas with shrubs, and not correlated with other agricultural landscapes (orchards and crop areas). However, the seroprevalence of T. gondii was only associated with agricultural landscapes such as orchards. The detection of hotspot areas of high pathogen exposure risk is the basis for municipal services to implement surveillance and risk factor control campaigns in specific-risk areas, including (a) efficient health management of urban cat colonies by geographical location, population census and health status monitoring of the components of each cat colony, (b) improvement of hygiene and sanitary conditions at the feeding points of the cat colony and (c) free-roaming cat trapping for health monitoring and, in the long term, to know the evolution of the health status of their populations.

Salmonella seroprevalence in wild boar from Southeast Spain depends on host population density.

Salmonellosis is the second most prevalent zoonosis in Europe and it has considerable economic and health implications for its monitoring and control as well as being among the most prevalent pathogens on livestock farms. The wild boar (Sus scrofa) has been identified as a competent host and spreader of Salmonella spp. There has been a significant increase in wild boar population in Europe in recent decades, and it is even present in urban areas. This study evaluates the spatial distribution of the seroprevalence of Salmonella spp. in wild boar from Murcia (Southeast Spain) and its relationship with host-related risk factors (sex, age, location and density). The presence of antibodies against S. Typhimurium and Choleraesuis in 269 serum of wild boars hunted in Murcia between 2015 and 2019 were analyzed using a commercial ELISA test (PrioCHECK porcine Salmonella kit). The seroprevalence were spatially distributed using Kernel function, and wild boar density using Gaussian kernel estimates (spatialEco version 1.1.1). The risk function was estimated as the ratio between the intensity of positive samples and the wild boar density The overall seroprevalence was 19.3% (IC95% 16.9-21.8), showing a significant spatial aggregation. The highest seroprevalence detected was 51.8% (IC95% 42.2-61.5) in a specific area with high risk of infection (76-100%) and was related to the wild boar density. Only marginal differences were detected for sex and age. The use of ELISA combined with QGIS (version 3.6.0) has allowed the identification of areas of Salmonella occurrence associated with high density as risk factor.

Disseminated bovine tuberculosis in a wild red fox (Vulpes vulpes) in southern Spain.

A wild-caught, adult red fox (Vulpes vulpes) from Doñana National Park (southern Spain), in very poor condition, died during anesthesia. At necropsy, the submandibular, retropharyngeal, mediastinal, axillary, mesenteric, and popliteal lymph nodes were enlarged, and the right submandibular and mesenteric lymph nodes had hard, white-colored concretions (calcifications). Multiple white pinpoint foci were observed in the lungs, and abscesses were present in the left dorsal lung lobes. No lesions were seen in tonsils, liver, or spleen. On histopathology multiple tuberculous granulomas, with Ziehl-Neelsen-positive bacilli, were observed in the lung, and multifocal necrotic areas with calcification were present in the submandibular and mesenteric lymph nodes. Nucleic acid amplification from lymph node samples demonstrated the presence of mycobacteria belonging to the Mycobacterium tuberculosis complex. One strain was isolated by culture and identified as Mycobacterium bovis. The suspected route of infection was oral, probably after repeated scavenging of infected wild ungulate carcasses. This is the first report of generalized bovine tuberculosis (bTB) in a wild canid. This finding raises concerns about bTB as a disease risk for protected species, livestock, and humans in Mediterranean Spain.

Pathogens of zoonotic and biological importance in roe deer (Capreolus capreolus): Seroprevalence in an agro-system population in France.

Antibody prevalence for several infectious and parasitic diseases in a population of roe deer (Capreolus capreolus) inhabiting a mixed agricultural landscape (south of France) has been analyzed. Serological analyses with ELISA in 245 animals captured from 2008 to 2012 has been performed. We found a high prevalence of Toxoplasma gondii (46.4%), Chlamydophila abortus (17.27%) and Coxiella burnetii (11.26%) compared to other studies in Europe. Seroprevalence varied strongly among years for T. gondii (27-91%), C. abortus (0-42%) and C. burnetii (0-27%). T. gondii prevalence was lower in juvenile females, compared to juvenile males and adults of both sexes. Other pathogens had low prevalences: Neospora caninum (1.56%), Bovine herpesvirus 1 (1.17%, 2008/09; 1.1%, 2010/11), Mycoplasma agalactiae (1.45%, 2009/10), Mycobacterium avium subsp. paratuberculosis (0.9%) and Slow viruses (CAEV-MVV) (0.15%, 2008/10; 0%, 2011/12). Antibodies to bluetongue virus and pestiviruses were not found in any individual.

Leptospirosis in wild and domestic carnivores in natural areas in Andalusia, Spain.

Leptospirosis is a zoonosis that affects humans, domestic animals, and wildlife. Carnivores are at the top of the feeding chain, thus being exposed to pathogens through their preys. From June 2004 to April 2007, we analyzed for evidences of contact with 14 serovars of Leptospira interrogans Sensu Lato serum (analyzed by indirect Microscopic Agglutination Test) and urine or kidney samples (analyzed by microscopic observation, immunostaining and culture) collected from 201 wild and domestic carnivores, including 26 free-living Iberian lynx (Lynx pardinus), 33 red foxes (Vulpes vulpes), 33 Egyptian mongooses (Herpestes ichneumon), 25 common genets (Genetta genetta), two Eurasian badgers (Meles meles) and one Eurasian otter (Lutra lutra), and 53 free-roaming cats and 28 rural dogs in protected areas in Andalusia (southern Spain). Twenty-three percent of the animals presented evidences of contact, being the prevalence similar among wild (23.5%) and domestic species (22.2%). Contact with Lesptospira was detected in all the species but the otter. Prevalence was: lynx (11% by bacteriological detection, 32% by serology), fox (0%, 47%), mongoose (5%, 20%), genet (0%, 12%), badger (0%, 50%), cat (20%, 14%), dog (only serology: 36%). Serovar Icterohemorragiae accounted for 2/3 of the cases. Serovar Canicola was detected in half of the positive dogs and one lynx. Other serovars detected were Ballum, Sejroë, and Australis. No macroscopic lesions were observed in necropsied animals that showed evidence of contact with the agent, although histopathologic lesions (chiefly chronic interstitial nephritis) were observed in 7 out of the 11 microscopically analyzed individuals. Thus, L. interrogans may cause previously unrecorded disease in wild carnivores in Spain. Wild and free-roaming carnivores may not act as reservoir of L. interrogans but as a dead-end hosts, though the dog may act as reservoir of serovar Canicola. Carnivores are apparently good sentinels for the epidemiological monitorization of leptospirosis.

Disease threats to the endangered Iberian lynx (Lynx pardinus).

The Iberian lynx, (Lynx pardinus), is the most endangered felid in the world. To determine whether sympatric carnivores are reservoirs of pathogens posing a disease risk for the lynx, evidence of exposure to 17 viral, bacterial and protozoan agents was investigated in 176 carnivores comprising 26 free-living lynx, 53 domestic cats, 28 dogs, 33 red foxes (Vulpes vulpes), 24 Egyptian mongooses (Herpestes ichneumon), 10 common genets (Genetta genetta) and 2 Eurasian badgers (Meles meles) in the areas inhabited by the last two populations of Iberian lynx, both in Andalusia (South-Western Spain). The results indicated that the lynx had low rates of contact with viral pathogens, with one seropositive finding each for feline leukemia virus, parvovirus and canine adenovirus-1, whereas contact with bacteria and protozoa appeared more frequent. Active infections with parvovirus, Ehrlichia spp., Mycobacterium bovis, Leptospira interrogans and Cytauxzoon spp. were confirmed. In contrast, 53% of the domestic cats were exposed to some infectious agent (prevalence range 4.5-11.4%). Antibodies to canine distemper virus and parvovirus were frequently found in dogs (32% and 42%, respectively) and foxes (30% and 12%). Past or present infections with parvovirus, Ehrlichia spp., Chlamydophila spp., M. bovis, Salmonella enterica, L. interrogans, Toxoplasma gondii, and Neospora caninum were also detected in these and other species surveyed. Questionnaires to owners revealed that 14% of the dogs but none of the cats had been vaccinated, and no cat had been neutered. Based on the apparent absence of acquired immunity of the lynx against infectious agents, the frequent detection of agents among sympatric carnivores, and the reported lack of immunocompetence of the Iberian lynx, a disease outbreak among the local abundant carnivores may pose a serious disease risk for lynx conservation.

Using a top predator as a sentinel for environmental contamination with pathogenic bacteria: the Iberian wolf and leptospires

The Iberian wolf (Canis lupus) is the top predator in the Iberian environments in which it lives, feeding on a wide range of species, thus encountering a wide range of disease agents. Therefore, the wolf can serve as sentinel of environmental contamination with pathogens. We investigated the exposure of free-living wolves to 14 serovars of Leptospira interrogans sensu lato. Kidney samples from 49 wolves collected from 2010-2013 in northwestern Spain were analysed by culture, direct immunofluorescence and polymerase chain reaction. Tissue fluids were analysed for antibodies by a microscopic agglutination test. Ten wolves (observed prevalence: 20%, 95% confidence interval = 11-33%) showed evidence of contact with leptospires, eight through direct detection and nine through serology (7 wolves were positive according to both techniques). Titres below the cut-off level were also detected in seven cases. Serovars confirmed were Canicola (n = 4), Icterohaemorrhagiae (n = 3) and Sejroë, Ballum and Grippotyphosa (n = 1 each), indicating that wolves were infected with serovars for which dogs, rodents and ungulates, are the natural hosts and supporting the utility of the wolf and other large predators as environmental sentinels for pathogens.