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In Europe wildlife animals such as the red fox (Vulpes vulpes) are considered the main reservoir for Angiostrongylus vasorum as well as a potential threat for domestic dog infection. Though this parasite is endemic in fox populations, data on A. vasorum infection in wolves (Canis lupus italicus) are still scant, having only recently been described in Northwestern Spain, in Italy, in Croatia and in Slovakia. Based on the rising number of cases of canine lungworm infection in Central Italy (Abruzzo region), the aim of the present study was to investigate the infection by A. vasorum in fox and wolf populations sharing the same geographical area of dogs. From October 2008 to November 2019, A. vasorum specimens were collected, through routine post-mortem examination, from 56 carcasses (44 foxes and 12 wolves). Adult parasites were searched for in the right side of the heart and in pulmonary artery of all carcasses. First stage of larvae (L1) was searched in faeces using the Baermann technique and in lungs by tissue impressions. Overall, 230 adult specimens were collected and identified on a morphological basis. To confirm the morphological identification, 4 adult specimens (n = 3 from fox, n = 1 from wolf) were molecularly identified as A. vasorum by amplification of partial fragment of nuclear 18S rRNA (~1700 bp) genes. The anatomo-pathological and parasitological examinations indicated the presence of A. vasorum in 33 foxes (75%) and in 8 wolves (66.7%). The level of prevalence of infested wolves was higher than the previous one reported in other European countries. Interestingly, the prevalence of infection in foxes herein recorded was higher than that described in dogs (8.9%) living in the same geographical area. This result may confirm the hypothesis that the spread of canine angiostrongylosis is linked to fox populations infection., Graphical abstract Image 1, Highlights • Angiostrongylus vasorum was found in dead foxes and wolves in Chieti province, Italy. • The prevalence in foxes was higher than that described in dogs. • The prevalence in wolves was higher than the previous one reported in European countries. • The spread of canine angiostrongylosis is linked to fox populations infection. • The province of Chieti appears to be an enzootic focus for canine angiostrongylosis.

The dire wolf was one of the successful top predators in North America during the Pleistocene. It is best known from the southern part of North America, and it even immigrated to South America. Fossils of dire wolves are very rare north of 42° North latitude in North America. That distribution supported the belief that the low temperatures and ice sheets in the higher latitudes of North American formed an insurmountable barrier for the dispersal of dire wolves. Here we report the first record of the dire wolf fossil in Eurasia. The fossil is a partial mandibular fragment with a tooth recovered from a Late Pleistocene underwater sand mine site near the city of Harbin in Northeastern China. Other mammalian fossils from the same site suggest that this dire wolf coexisted with the typical Mammuthus-Coelodonta fauna of Eurasia. The newly discovered specimen has a huge m1; much larger than gray wolves and other large canids from the same region. The massive m1 trigonid accounts for a high percentage of its overall length. The m1 talonid is reduced, but retains a small entoconid. The m2-3 alveoli suggest that the two teeth were smaller than those of the gray wolf. The combination of these morphologies is present only in the dire wolf. When the body mass spectra of medium and large-bodied carnivores from Asia and North America were compared, the Asian dire wolf occupies a position nearly overlapping the spotted hyena (Crocuta crocuta ultima). During the Late Pleistocene, C. crocuta ultima was one of the most widely distributed and dominant carnivores in Asia. Competition from hyenas may have kept dire wolf Asian populations at very low levels leading to their rarity as fossils rare in Asia.

Development of genomic preservation technologies for canids, especially for seasonally breeding species like the grey wolf (Canis lupus), is needed in advance of growing species conservation concerns. Here, we evaluated the efficacy of two cryopreservation protocols – needle immersion vitrification (NIV) and slow freezing (SF) on grey wolf (n = 7) testicular tissue morphology. NIV samples were equilibrated in a 7.5% v/v dimethyl sulfoxide (DMSO or Me2SO) + 7.5% ethylene glycol (EG) solution in minimum essential medium with 20% FBS for 10 min at 4 °C, then exposed to 15% DMSO + 15% EG + 0.5 M sucrose for 10 min at 4 °C before plunging into liquid nitrogen. For slow freezing, we assessed two cryoprotectant (CPA) strategies, DMSO, 15% v/v alone (SF-D) or 7.5% EG + 7.5% DMSO (SF-ED). Following thawing, there were no significant differences in seminiferous tubule area among treatment groups, although all cryopreserved tissues displayed reduced tubule size compared with fresh controls and increased apoptosis, the latter reaching significance for SF-D treated tissues. Slow freezing improved maintenance of testis architecture, with minimal detachment of seminiferous tubule basement membranes post-thaw. Spermatogonia densities were reduced in NIV tissues compared with fresh, with no differences in spermatocyte, spermatid, or Sertoli cell counts, or germ cell marker DDX4+ cell densities among groups. In sum, we conclude that slow freezing better maintained morphology of cryopreserved testicular tissues compared with needle vitrification with 15% each DMSO and EG and 0.5 M sucrose, and that DMSO + EG combination SF supports cell viability. This represents a first step in the development of male gonadal tissue preservation strategies for the grey wolf.

The aim of the study was to extend the knowledge about to the total locomotor activity behavior in maned wolves (Chrysocyon brachyurus) through the comparison with domestic dogs (Canis familiaris). Five maned wolves housed individually in 4 m2 dens with nest boxes and runs allowing them to move in outdoors area of 250 m2 with natural vegetation; and five Beagles dogs living individually in a 12 m2 box with an outside area of 50 m2 were subjected to natural light-dark (L/D) cycle (maned wolves: 15 L/9D; dogs 14 L/10D) and environmental conditions (maned wolves: 22-26°C, 64Rh%; dogs 17-23°C, 65Rh%). In each animal, the monitoring of locomotor activity was performed for 2 days by means of an activity data logger (Actiwatch Cambridge Neurotechnology, Cambridge, UK), applied on a neck collar. Data were analyzed by the single Cosinor method. At the obtained rhythmic parameters, two-way analysis of variance was applied to verify differences due to day of monitoring and species. Maned wolves showed a nocturnal daily rhythm of locomotor activity. Dogs showed a diurnal daily rhythm of locomotor activity. The diurnality index was 0.29 and 0.93, respectively, for maned wolves and dogs. The two studied species showed statistically difference in mesor and amplitude of rhythm, higher in maned wolves (mesor: 3256.64 ± 364.87 arbitrary unit; amplitude: 3726.32 ± 327.50 arbitrary unit) than dogs (mesor: 414.06 ± 9.68 arbitrary unit; amplitude: 502.35 ± 18.78 arbitrary unit). We can claim that maned wolves, housed in captivity are more active than dogs and showed an opposed daily rhythm of locomotor activity.

Reports in the literature indicate that species of Hepatozoon commonly occur in African wild dog (AWD) or painted wolf (Lycaon pictus) populations. These findings were based on examination of blood smears by microscopy, and specific identity of the Hepatozoon sp. gamonts seen could not be confirmed. We present the first in-depth molecular data on the prevalence of species of Hepatozoon in a free-ranging AWD population. In a general health survey of AWDs in the Kruger National Park, blood specimens (n = 74) collected from 54 individuals were examined for the presence of Hepatozoon spp. At first sampling, specimens from 42 of 54 individuals (77.7%) were positive, based on the primer set HepF300 and HepR900. Twenty individuals were resampled between 51 and 69 days after first sampling; one of these was resampled twice. Samples from six individuals that had tested negative previously now reacted positive. Assuming that all 54 individuals were still alive, the prevalence had therefore increased to 48 individuals infected, or 88.8%. Resultant 18S rDNA sequences isolated from these specimens share high similarity to other Hepatozoon canis genotypes. Phylogenetic analysis recovered the Hepatozoon sp. isolated from AWDs within the H. canis cluster, which includes species of Hepatozoon from other canid and tick hosts.

Dispersal is a fundamental ecological process that, on an individual level, is associated with intrinsic and fixed personality traits such as ‘boldness’. However, it is unknown whether personality traits or behavioural syndromes might change as a consequence of dispersal itself. We analysed 14 GPS-collared grey wolves, Canis lupus, in Finland that dispersed from their natal territories and settled in new territories, as well as 22 time-matched nondispersing controls. We used wolf avoidance of low-use forest roads, high-use primary roads and houses as proxies for boldness, and we estimated wolf responses to these features across spatial scales and across dispersal phases. Wolves, which already occupy portions of Finland with low human density, expanded into more human-impacted areas, but with a strong large-scale selection for relatively low human presence. At the finer scale, avoidance of human elements varied as a function of dispersal phase. During dispersal, avoidance of forest roads, primary roads and houses was substantially lower than in natal territories, reflecting the need to take more risks while moving in a small group (alone or with a single mate) in unfamiliar environments. After settling in new territories, the strength of avoidance of those elements continued to be lower than in natal territories, even after controlling for changes in availability. These results suggest that experiences gained across a major life-history transition, such as dispersal, might themselves influence personality traits.

Wild ungulates can recognize certain predators without previous experience, but this innate ability may be relaxed under domestication. Using naive dairy calves, Bos taurus, we examined the effects of exposure to a predator odour (coyote, Canis latrans, urine) and two control odours (deer urine and water) on (1) latency to approach a milk food reward, (2) exploration, vigilance and locomotor play, (3) magnitude of the startle response to a sudden noise delivered upon arrival at the feeder and (4) heart rate during feeding in 10 min trials on three consecutive days. In response to predator odour, calves delayed feeding and tended to explore more and play less in the first 30 s of the trial compared to calves exposed to control odours. The duration spent feeding and standing with the head raised (i.e. vigilant) did not differ among treatments. On the first day, calves exposed to predator odour startled more to a sudden noise than control animals. Mean heart rate during feeding decreased across days in the control treatments, but not in the predator treatment, suggesting a sustained response to the predator odour. We conclude that naive domestic calves retain the ability to distinguish between predator and nonpredator odour.

Toxocariasis, a natural helminth infection of dogs and cats caused by Toxocara canis and T. cati, respectively, that are transmitted to mammals, including humans. Infection control is based currently on periodic antihelmintic treatment and there is a need for the development of vaccines to prevent this infection.Eight potential vaccine candidate T. canis recombinant proteins were identified by in silico (rTcGPRs, rTcCad, rTcVcan, rTcCyst) and larval proteomics (rTES26, rTES32, rMUC-3 and rCTL-4) analyses. Immunogenicity and protection against infectious challenge for seven of these antigens were determined in a murine model of toxocariasis. C57BL/6 female mice were immunized with each of or combinations of recombinant antigens prior to challenge with 500 T. canis embryonated eggs. Levels of specific antibodies (IgG, IgG1, IgG2a and IgE) in sera and cytokines (IL-5, INF-ɣ and IL-10) produced by antigens-stimulated splenocytes, were measured. Presence of specific antibodies to the molecules was measured in sera of T. canis-seropositive dogs and humans.All seven molecules were immunogenic in immunized mice; all stimulated significantly elevated levels of specific IgG, IgG1 or IgG2a and six were associated with elevated levels of specific IgE; all induced elevated production of IFN- ɣ and IL-10 by splenocytes, but only the in silico-identified membrane-associated recombinants (rTcCad, rTcVcan, and rTcCyst) induced significantly increased IL-5 production. Vaccination with two of the latter (rTcCad and rTcVcan) reduced larval loads in the T. canis challenged mice by 54.3% and 53.9% (P 0.0001), respectively, compared to unimmunized controls. All seven recombinants were recognized by T. canis-seropositive dog and human sera.The identification of vaccine targets by in silico analysis was an effective strategy to identify immunogenic T. canis proteins capable of reducing larval burdens following challenge with the parasite. Two recombinant proteins, rTcCad and rTcVcan, were identified as promising vaccine candidates for canine toxocariasis.

Football matches, as a common urban activity, produce high levels of noise because of vehicular traffic, screams, whistles, and firework displays. Fear of noise is one of the most commonly reported behavior problems in dogs (Canis familiaris); however, few studies have focused on the effects of repeated exposition to noise in these animals. Here, we investigated whether the noise produced during football matches may act as a stress source for dogs living near football stadiums. We measured noise levels on days with and without matches, in the surrounding neighborhoods of two football stadiums of Minas Gerais, Brazil: (a) Arena Independencia (AI) and (b) Governador Magalhaes Pinto (GMP). Noise measurements were carried out by using a sound level meter, in distances varying between 0 and 400 m from the stadiums. A structured interview was conducted to dog owners living nearby AI, about the dogs' characteristics, demographics, and behavior. Noise records of days with and without matches were compared, and questionnaires were analyzed (general linear models) to identify possible correlations between the reported changes in the animals’ behaviors on days with football matches and the distance these animals lived from the stadium. Higher noise levels were recorded on football match days than on days without matches, with a greater effect of these between 0 and 249 m (D1) than between 250 and 400 from AI. Around Governador Magalhaes Pinto, we recorded greater noise levels on days without football matches compared with match days, with no difference between distances. Fear/anxiety behaviors were reported as more intense on days with football matches, in comparison with days without matches for 90.6% of the study dogs. The occurrence of matches made specific fear-related behaviors of the dogs—severe trembling, excessive salivation, agitation/restlessness, whining, and appetite loss—more frequent. The proximity of AI intensified the exhibition of general fear/anxiety behaviors and made intense trembling and excessive salivation more frequent at D1, in relation to D2. On the other hand, dogs living at D2 barked, howled, and lost appetite more often than those living at D1. Our results suggested a detrimental influence of noise from football matches on the behavior of dogs living around football stadiums. These data are relevant for public policy-making to promote preventive and mitigating measures, to improve the quality of life of both the dog and the human populations in the surroundings of football stadiums.

Helminths have developed complex mechanisms to suppress the host immune response. These mechanisms may impair the host vaccine response. This study aimed to evaluate the effect of Toxocara spp. infection on the vaccine immune response to bovine herpesvirus type 5 (BoHV-5). First, 30 heifers received two doses of an experimental BoHV-5 vaccine. At 42nd days after the primo vaccination the vaccine efficacy was evaluated, and the presence of anti-Toxocara antibodies. Second, 20 Balb/c mice were divided into two groups, one infected with T. canis and the other without infection. After infection, both groups received two doses of vaccine. The vaccine immune response was assessed by BoHV-5 serum neutralization and splenic cytokines transcription by qPCR. All heifers positive for Toxocara spp. (40%) showed BoHV-5 SN titer ≤1:32, whereas heifers negative for Toxocara spp. (60%) had BoHV-5 SN titer ≥1: 128. Infected T. canis mice showed BoHV-5 SN titer ≤1:2, whereas mice not infected with T. canis BoHV-5 SN titer ≥1:8. Splenocytes from control mice stimulated with BoHV-5 had a significant (p .05) mRNA transcription for the cytokines IL-12, IL-17, and IL-23, whereas the same cytokines were down-regulated in T. canis infected mice. These results suggest that Toxocara spp. infection may impair BoHV-5 immunization and should be considered for efficient herd immunization.

Background Toxocariasis is an infection caused by larvae of a group of animal nematodes named Toxocara in human tissues. The disease often affects children under the age of 10 years. In this research, we decided to determine the prevalence of infection in schoolchildren of Bushehr city as a first study in the region. Methods In this cross-sectional study, the samples were selected randomly among Bushehr primary school students between September 2011 and June 2012. ELISA method was used to measure anti-Toxocara canis IgG antibodies. Data were collected by questionnaire and analyzed using SPSS 18.0 software and Chi square test. Results Of the 400 students studied, 48 (12%) were positive for anti-Toxocara IgG antibodies. Results showed that there is no a statistically significant relationship between infection and some of the analyzed variables such as sex, abdominal pain, anorexia, chronic cough, diffuse pain, itching and skin sensitivity. However, with the habit of playing with soil, this relationship was significant. Conclusion Considering the importance of the disease in children and influence of harmful effects on their health, it is suggested that the methods for diagnosis be trained for clinicians in the area and periodically tests as well as teaching the prevention and control strategies should be considered to the educational staff and parents.

The Toxocarosis is a common zoonotic parasitic infection caused by three etiological agents T. cati, T. canis and T. vitulorum. It has a severe impact on health and economy especially in countries where it is endemic. The study was conducted from June 2013–2014. The blood samples were collected from (n = 270) cattle grazing in the Potohar region. The sera were separated and subjected to anti-Toxocara ELISA and results revealed an overall seroprevalence of 16.67% (45/270) in cattle. The relationship between age groups and seroprevalence of T. vitulorum indicated a statistically significant difference (p = .01, χ2 = 16.81) and non-significant was sex group of cattle and seroprevalence of T. vitulorum (p = .859, χ2 = 0.032). The results also revealed a statistically non-significant difference between cattle breed and Toxocara infection (p = .142, χ2 = 5.424). From the study, we concluded that indirect ELISA is a highly efficient assay for timely diagnosis of infection in order to adapt control strategies to mitigate the infection. The present study recommended the need to develop ELISA by using excretory-secretory (ES) T. vitulorum antigen in order to reduce the cost of commercial kits for mass screening purposes. Furthermore, the result of the current study also suggested and conducting similar nature of studies in other agroecological zones of Pakistan, which will help to provide baseline epidemiological data of parasitic diseases.

Cardiopulmonary nematodes are among the most pathogenic parasites of domestic and wild canids. The aim of this study was to describe the species diversity, prevalence and infection intensity of these parasites in the Iberian wolf (Canis lupus signatus) and the red fox (Vulpes vulpes) in the northwest of the Iberian Peninsula. 257 foxes and 74 wolves were necropsied between 2008 and 2014. Four nematode species were identified: Angiostrongylus vasorum, Eucoleus aerophilus, Crenosoma vulpis and Filaroides hirthi. This last species was only found in wolves, being the first time that is cited worldwide in this wild canid. The overall parasite prevalence was significantly higher in foxes (70%) than in wolves (28%). Specifically, prevalences in foxes and wolves were, respectively, 43% and 22% for A. vasorum, 33% and 5% for E. aerophilus, and 30% and 9% for C. vulpis. The prevalence of F. hirthi was 16%. The A. vasorum intensity was significantly higher in foxes than in wolves. Differences between host species in the risk of infection would be associated to diverging feeding behavior, and possibly reflects a parasite-host adaptation related to host's hunting strategies and cardiorespiratory requirements. This study revealed an association between infection and environmental factors, and highlighted a wide variation in the spatial distribution of A. vasorum. Our results indicate that cardiopulmonary parasites are widespread in wild canids in northwest Spain, and further agrees with other studies indicating the expansion of A. vasorum in Europe and, therefore, the urgent need to investigate infection in dogs in sympatric areas.

In some cooperatively breeding species, breeding and nonbreeding individuals within a group care for young through provisioning and guarding, commonly referred to as alloparental care. The question of why nonbreeders contribute to the care of young is at the centre of research on the evolution of cooperative societies, and the mechanisms that drive the behaviour are a topic of debate. We employed a novel approach to examine grey wolf, Canis lupus, space use at pup-rearing sites by using genotyped scats to measure the distance between wolves and evaluate the influence of genetic relatedness and other group characteristics on this space use. Specifically, we evaluated how group composition and individual characteristics influenced space use and whether the distance between adults and pups influenced pup survival. We genotyped 3093 wolf scats collected at pup-rearing sites during June–August 2008–2016 and calculated the distance between scats. We found that space use at pup-rearing sites did not influence pup survival. We also found that nonbreeders, especially yearlings, remained closer to pups than did breeders, and that individuals more genetically related to the pups were found closer to them, suggesting kin-directed attendance behaviour. Finally, we found evidence suggesting that direct fitness benefits, such as gaining access to food, may influence space use as well and help to explain why yearling nonbreeders were found the closest to the pups. This is the first study to provide indirect evidence for kin-directed attendance behaviour in wolves and it contributes novel methodology for investigating space use and helping behaviour in cooperative breeders at reproductive sites.

Classification plays a pivotal role in our attempts to develop an understanding and expectation of animal behavior. The hypothesis underpinning traditional breed group classification of domestic dogs is that behavioral differences among breeds may be explained by selection to perform particular roles. Consequently, breed group membership may provide an explanation of differences in performance in problem-solving tests. This study examined owner-reported performance of 8,063 pedigree dogs of three problem-solving tests designed to assess performance of different aspects of animal cognition. We asked (1) whether there are significant differences in problem-solving performance between members of breed groups in their performance of 3 cognitive tests and (2) the utility of breed group profiles in providing an explanation for the highest performing breeds. Results indicate that counter to popular perception of breeds as distinct populations manifesting differing and predictable “breed-group typical” cognitive abilities, the findings revealed no such differences in problem-solving performances between breed groups although there was variation between breeds. High-performing breeds could not be explained by a particular morphology type or original breed origin function as reflected in Kennel Club classifications. It is suggested that breed group classifications are inadequate in yielding useful explanations of problem-solving performance. The results caution against an overreliance on such classifications for understanding dog behavior.

Urban wildlife populations present different scenarios for managers compared to rural populations, partly because of greater diversity in stakeholder attitudes and opinions regarding urban wildlife. Wild urban canids—especially coyotes (Canis latrans)—have been of increasing interest throughout North America in recent years. Our objective was to evaluate the potential of using iNaturalist-generated observations of urban red foxes and coyotes for cost-effective, customizable data collection to inform urban canid management. Our research is important on two fronts; first, it is a way to engage the public to make them more aware of urban canids, and secondly, it is an attempt to empirically test if we can more efficiently and effectively track coyotes and red foxes in urban areas. We used iNaturalist to collect over 800 community-generated locations for red foxes (Vulpes vulpes) and coyotes in Madison, WI from 2015 and 2016. We concurrently placed radio-collars on 11 red foxes and 11 coyotes to determine areas used in this urban ecosystem. We compared iNaturalist to radio-telemetry locations to identify factors that led to a positive relationship between these two inherently different spatial data sets. Greatest overlap between iNaturalist and telemetry data for both red foxes and coyotes occurred in areas with moderate human development and there was minimal overlap in natural areas. The overlap between iNaturalist and telemetry locations was comparable for both species, but the underlying mechanism differed by species-specific habitat use. iNaturalist reports appeared to show where and when humans most often interacted with red foxes and coyotes, rather than their true spatiotemporal distribution. Understanding the relationship between community-generated reports and local canid distribution may inform how iNaturalist can be used as a management tool and allow managers to proactively monitor and manage human-wildlife interactions with urban wildlife.

Unawareness of canine parasitic diseases among at-risk hosts and an uncontrolled program of stray dog population have caused that zoonotic parasites received great attention in endemic regions of the Middle East. A total of 552 faecal samples were collected between December 2016 to January 2018 from stray (n = 408) and domestic (n = 144) dogs of Iran. All specimens were coproscopically observed following concentration and flotation techniques. Subsequently, the DNAs of taeniid eggs were extracted, amplified, and sequenced by targeting of mitochondrial cytochrome oxidase subunit 1 and small-subunit ribosomal DNA markers. The overall prevalence of canine intestinal parasites found 53.6%. The following parasites and their total frequencies were identified: taeniid (10.5%), Dicrocoelium dendriticum (0.7%), Trichuris vulpis (1.2%), Capillaria spp. (2.3%), Blastocystis spp. (5.2%), Ancylostoma spp. (2%), Eimeria spp. (13.2%), Dipylidium caninum (2.3%), Toxocara canis (3.8%), Giardia spp. (8.5%), and Toxascaris leonina (3.6%). Stray dogs were characterized more likely to be poliparasitized and indicated a higher prevalence of taeniid (10.9%), T. canis (4.4%) Giardia spp. (10.1%) than domestic dogs (P > 0.05). Phylogenetic and sequence analysis of Cox1 and SSU-rDNA indicated a low genetic diversity (Haplotype diversity; 0 to 0.495) in E. granulosus sensu lato G1, G3, G7 genotypes, and Taenia hydatigena. The pairwise sequence distances between G7 isolates showed an intra-diversity of 0.7%–1.5% and identity of 98.5%–100%. The first occurrence of pig strain (G7) from Iranian dogs might have substantial implications in the drug treatment of infected dogs due to the shorter maturation time of G7 compared with G1 genotype. Thus, the preventive strategies should be noticed to determine the risk factors, the importance of applying the hygienic practices, and well adjusting deworming programs for the Iranian dogs and at-risk individuals.

Infectious haemolytic anaemia (IHA) in dogs share similar clinical signs including fever, lethargy, icterus, paleness of mucous membranes and splenomegaly. Postmortal findings are similar and, without additional diagnostic methods, an accurate aetiological diagnosis is difficult to achieve. In order to investigate causes of lethal IHA in Croatian dogs, we performed a retrospective study on archived formalin-fixed, paraffin-embedded tissue blocks (FFPEB) from dogs that died due to haemolytic crisis, using microscopic and molecular diagnostic tools to determine the aetiological cause of disease. Molecular analysis was performed on kidney, lung, myocardium and spleen on FFPEB from all dogs. The originally stated aetiological diagnosis of B. canis or leptospirosis was confirmed in only 53% of the dogs. PCR and sequencing revealed that, in addition to the expected pathogens, B. canis and Leptospira interrogans, the presence of previously undiagnosed "new" pathogens causing anaemia including Candidatus Neoehrlichia mikurensis and Anaplasma phagocytophilum. Furthermore, Theileria capreoli was detected for the first time in a dog with postmortal descriptions of lesions. Intensive extravascular hemolysis was noticeable as jaundice of the mucosa, subcutis and fat tissue, green or yellow discoloration of renal parenchyma caused by bilirubin excretion in the renal tubules and bile accumulation within the liver in 90% of the dogs. This work highlights the value of molecular diagnostics to complement traditional ante-mortem and post-mortem diagnostic protocols for the aetiological diagnosis of pathogens associated with IHA.

Livestock depredations present real and perceived threats to property and human livelihood, undermining carnivore conservation and management. Considerable research has been aimed at identifying factors influencing depredations but contradictory relationships indicate our predictive ability still needs improvement. Previous approaches have relied primarily on linear correlation modeling not considering that some factors may promote and limit depredations across the range of observed conditions. We investigated relationships between wolf (Canis lupus) depredations of livestock and indices of human activity in Michigan’s Upper Peninsula (UP), USA. Using binomial generalized linear models at three spatial scales (site scale, radius [r] = 0.9 km; area of mean 50% wolf core area [core area scale], r = 4.4 km; and 95% wolf territory area [territory scale], r = 9.0 km), we tested the hypothesis that the relationship between depredation probability and indices of human activity is nonlinear, and that the greatest probability of depredations occurs at an intermediate level of conditions. Across spatial scales we found support for quadratic relationships between cattle density, human density, and proportion of agricultural lands with the occurrence of wolf-livestock depredations (n = 260). We also demonstrated that at the wolf territory scale, increased road density reduced the probability of depredations. Using test data, model prediction accuracy for estimating depredations was 90% (n = 28) at the land section scale and 84% (n = 26) at the core area scale. We provide demonstrative use of non-linear modeling to evaluate factors influencing depredations robust to wolf and prey distribution and abundance.

Background Canine brucellosis , caused by the bacterium Brucella canis , is a zoonotic and largely reproductive disease of dogs. The disease is a recognized problem in canine breeding populations, and the risk to individuals assisting with birthing is well described. Prior to 2015, all cases of canine brucellosis reported to the Minnesota Board of Animal Health were in dogs used for breeding. In 2015, canine brucellosis was identified in eight Minnesota rescue dogs, all originating from specific geographic areas in South Dakota. Our objective was to measure the seroprevalence of B. canis in stray and previously owned dogs entering a large Minnesota animal rescue organization to determine if our observations represented a localized or generalized disease issue among rescue dogs. Methods A stratified random sample of stray and previously owned dogs entering the largest Minnesota animal rescue organization between November 1, 2016 and November 7, 2017, was tested for B. canis antibodies by the 2-Mercaptoethanol Rapid Slide Agglutination Test (2ME-RSAT) (Zoetis d -TEC® CB kit). Sample sizes for each strata were calculated using previously published seroprevalence estimates. Blood from selected dogs was collected, serum harvested, and transported to the Minnesota Veterinary Diagnostic Laboratory for testing. Positive samples in the 2ME-RSAT were shipped to Cornell University for confirmation by Agarose Gel Immunodiffusion (AGID) testing. Demographics, state and setting of origin, and health status were collected on study-dogs. Results Of the 10,654 dogs accepted by AHS during the study period, 943 (8.9%) were selected for testing. Most study dogs arrived from Oklahoma (28%), Alabama (18%), and Minnesota (12%). The median age of study dogs was 1.5 years; 303 (32%) were intact males and 294 (31%) were intact females. Most study dogs were strays (n = 716, 76%). Of the total, 22 (3.1%) stray and eight (3.5%) owner-surrendered dogs were presumptively positive by RSAT; one (0.11%) of the stray dogs was positive by 2ME-RSAT and confirmed by AGID. The positive dog was a healthy-appearing 1 year-old neutered male beagle from Texas. Conclusions The seroprevalence of canine brucellosis in dogs entering Minnesota for adoption from multiple states was low. Never-the-less, care must to be taken to consider all potential risks and outcomes of interstate and international dog trade, including the spread of infectious diseases such as canine brucellosis.

Domestic dogs, Canis familiaris, are highly responsive to human communicative cues and can utilize gestures, such as pointing, to locate hidden rewards. This ability is thought to be the product of both genetic and behavioural selection, allowing dogs to adapt to life with humans. Dogs' responsiveness to human gestures can also lead to suboptimal choices when dogs readily follow misleading cues despite directly contradicting perceptual information such as odour cues. However, this bias likely reflects pet dogs' enculturation with humans and thus may not be representative of other populations of dogs. We investigated the ability of young dogs in training for explosives detection (N = 77) to locate a hidden reward using olfactory cues when presented in conflict with a deceptive communicative gesture in an object-choice task. We assessed performance at 3, 6 and 11 months of age using both cross-sectional and longitudinal designs. We found that, contrary to previous findings with pet dogs, responsiveness to human pointing decreased with age, whereas the ability to locate the reward by scent increased. Furthermore, a lack of susceptibility to deceptive social cues was predictive of future success as a detection dog. These findings further indicate the influence of ontogenetic effects on canine social cognition and demonstrate potential applications for the identification of suitable detection dogs.

An unexpected change in reward quantity or quality frequently elicits a sharp decrease of responses as well as a negative emotional state. This phenomenon is called successive negative contrast (SNC) and, although it has been observed in numerous mammals, results in dogs have been inconsistent. The aim of this study was to evaluate SNC in dogs, comparing the effects of rewards of different qualities in a non-social task carried out in the dogs? usual environment. Dogs were separated into two experimental groups that experienced a downshift from a high quality reward (liver or sausage) to a low quality one (dry food), as well as a control group that always received dry food. The task involved a dog toy with bone shaped pieces that had to be removed to get the food hidden underneath. When the reward changed from liver to dry food, dogs picked up significantly fewer bones than the control group. However, this effect was not observed with sausage. Results show SNC in dogs in a non-social task carried out in their home environment. Additionally, the importance of the discrepancy in the hedonic value of the rewards is highlighted. Fil: Dzik, Marina Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina Fil: Cavalli, Camila María. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina Fil: Iglesias, M.. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina Fil: Bentosela, Mariana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; Argentina

Following decades of absence, the gray wolf (Canis lupus) has recolonized much of the northern Great Lakes region from Canada and remnant populations in northern Minnesota. The wolf population in Michigan’s Upper Peninsula may now be reaching saturation, with evidence that some dispersing individuals have traversed the Straits of Mackinac during ice-over winter conditions indicating potential recolonization of northern Lower Michigan. While previous research suggests suitable habitat exists in northern Lower Michigan to support a small wolf population, habitat availability at other hierarchical levels, including den habitat and the ability of individuals to disperse successfully among suitable habitat patches, has not been assessed. We evaluated the den habitat availability and landscape connectivity using a multi-scale modeling approach that integrates hierarchical habitat selection theory as well as spatial structure to assess whether corridors exist for wolves to successfully recolonize and raise pups in northern Lower Michigan. We used expert opinion, scientific literature, and geographical information systems to develop models of landscape suitability, resistance, and least-cost path analysis to identify dispersal corridors throughout the Upper and northern Lower Peninsulas of Michigan. Based on our models, the Upper Peninsula was almost entirely amenable to wolves for both denning and dispersing, particularly in the western portion of the peninsula. Our estimates indicate that over 1900 km2 of high quality den habitat exists in northern Lower Michigan, but landscape permeability between these habitat patches appeared relatively low relative to Upper Michigan. We delineated several corridors of high quality habitat in the Upper Peninsula that may facilitate dispersal in to Lower Michigan. Dispersal corridors were of moderate quality in northern Lower Michigan, representing higher mortality risk but potentially capable of promoting recolonization of high-quality habitat areas. Conservation efforts within these identified corridors may further increase the potential for successful recolonization and establishment of viable long-term breeding populations of gray wolves in northern Lower Michigan.

The ability to determine body mass from skulls is valuable for understanding various ecological, physiological, and evolutionary factors. In the Canidae, numerous methods to reconstruct body mass from measurements of the skull have been proposed, however there is no one-size fits all approach that can be applied across all species and subspecies. Added to this, current methods of reconstructing body mass are often complex, and have relatively high error rates. We aimed to produce a multivariate regression equation to estimate body mass of the Australian dingo (Canis dingo) from simple measurements of the skull, whilst ensuring that it could also be used in studies of encephalisation. To do this, we focussed on palate length (PL), palate width (PW) and the length of the first upper molar (M1). A total of 128 adult dingo (64 male; 64 female) crania from one region of Australia with known body mass were measured. Overall, the combination of PL and PW was the best predictor of body mass, with M1 having poor predictability. The model, mass (kg) = 0.246 ∗ (PL) + 0.320 ∗ (PW) − 24.757 produced a prediction error of 8.05%. Thus, these two measures of the palate provide simple and accurate predictors of body mass for the dingo. This will be useful for modern dingo specimens, as well as those found at archaeological sites and in museum collections that often consist of incomplete cranial material. The reconstruction of dingo body size is useful for evaluating variation in body mass through time, and across the Australian continent, particularly in the context of human activity.

Evidence suggests that paw preferences are related to emotional functioning in the domestic dog. Whether motor bias in this species is related to the display of behaviour problems, which often have their basis in emotional reactivity, is still unknown. This study therefore explored the relationship between lateralised motor behaviour in the form of paw preferences and the occurrence of canine behaviour problems. Fifty-two dogs presenting with one or more behaviour problems, and 61 control animals, had their paw preferences assessed using the commonly employed Kong™ ball test. The dogs’ behavioural profile was determined using the C-BARQ, a psychometric tool designed to provide standardized evaluations of canine behaviour. Analysis revealed a roughly equal split in the proportion of dogs classified as ambilateral and lateralised, although dogs were more like to be ambilateral than left- or right-pawed. Dogs differed significantly in their behavioural profile, with animals in the behaviour problem group displaying more severe patterns of stranger- and dog-directed aggression and fear, non-social fear and touch sensitivity than those in the control group. There was no significant difference in the distribution, direction or strength of the dogs’ paw use between the two groups of animals. The dogs’ paw preference classification was significantly associated with one of the C-BARQ subscale scores, namely stranger-directed aggression, with left-pawed dogs having lower scores on this subscale than right-pawed or ambilateral animals. The direction of the dogs’ paw use in the behaviour problem group was significantly correlated with C-BARQ scores for the subscales of stranger-directed aggression, stranger-directed fear and attention-seeking behaviour, with higher subscale scores associated with increasing right-pawedness. There was no significant correlation between the direction of the dogs’ paw use and any of the other C-BARQ subscale scores. The strength of the dogs’ paw preferences was not significantly correlated with any of the C-BARQ subscale scores for either the control or behaviour problem group of animals. Canine sex was not significantly related to either the dogs’ paw preferences or the presentation of behaviour problems. Overall, this study provides little evidence in support of a relationship between paw preference and the occurrence of behaviour problems in the domestic dog. Further, longitudinal, work is recommended in an effort to unravel the ontogeny of lateralisation and its association with emotional reactivity in this species.

This study examines the activity budgets and social behaviours initiated and received by 69 focal dogs in an off-leash dog park for 400 s after entry, a time of high activity about which little is known. Using motivationally-neutral labels for social behaviour categories, we describe the frequency of behaviours, and correlations among them. We then examine these relationships in the context of proposed functions for some behaviours in dogs, in terms of information gathering and communication, including visual and tactile signalling. Time spent with other dogs decreased rapidly over the visit, and much of this early interaction involved greeting the park newcomer. Snout-muzzle contact behaviours were ubiquitous, while other behaviours were rarely observed, including aggressive behaviours. Correlations among certain non-contact behaviours initiated and received by focal dogs are consistent with their function as visual signals that may influence the continuation and form of social interactions, and their possible role in social mimicry (i.e., play bow and pull-rear away). Age, sex, and number of dogs present in the park influenced specific aspects of dogs’ activity budgets, and a few behaviours. This ethological study provides fundamental data on dog social behaviour in dog parks, about which surprisingly little has been published.

Data concerning combined molecular and serological prevalence of emerging canine tick-borne pathogens in Serbia are lacking. A large population of outdoor living dogs in Belgrade, Serbia's' capital, present an excellent population for epidemiology study. Blood samples were collected from 111 dogs, including 46 shelter, 31 free roaming, and 34 hunting dogs. Species-specific real-time polymerase chain reaction (PCR) (IDEXX Laboratories, Inc., Westbrook Maine, USA) was applied for the molecular detection of Anaplasma phagocytophilum, A. platys, Ehrlichia canis, Babesia canis, B. gibsoni and B. vogeli. A research based SNAP assay (SNAP® M-A, IDEXX Laboratories, Inc., Westbrook Maine, USA) that uses genus and species-specific peptides was used to asses Anaplasma spp., A. phagocytophilum, A. platys, Ehrlichia spp., E. canis, E. chaffeensis, E. ewingii and Borrelia burgdorferi antibody status. B. canis, B. gibsoni and B. vogeli antibody status was assessed with an indirect immunofluorescence test (MegaCor Diagnostic, Horbranz, Austria). Anaplasma spp. and Ehrlichia spp. DNA was not amplified. One quarter of the dogs were A. phagocytophilum, one dog was A. platys, one was E. ewingii and two dogs were B. burgdorferi seroreactive with the SNAP® M-A. Babesia canis or B. gibsoni DNA was amplified by PCR from 16.2% of dogs, whereas 67.6% were seroreactive to one or more Babesia spp. Babesia vogeli was not PCR amplified. We conclude that outdoor dogs in this territory are reservoirs for B. canis and B. gibsoni and are frequently co-exposed to combinations of Anaplasma and Babesia spp.

Management of wildlife often relies upon understanding mechanisms linking anthropogenic disturbance to population declines. The most-cited mechanism by which disturbance threatens boreal caribou (Rangifer tarandus caribou) is the exacerbation of apparent competition via increases in early successional forage, and subsequent changes in the densities and distributions of other prey species and gray wolves (Canis lupus). An alternative mechanism is the direct alteration of wolf distribution via positive responses by wolves to anthropogenic features. We conducted a mechanistic evaluation of hypotheses explaining human-mediated increases in boreal caribou mortality across northeast British Columbia. We evaluated support for (i) numeric apparent competition (increased prey densities) by evaluating relationships between disturbances, moose (Alces alces) density, and caribou survival. To evaluate (ii) spatial apparent competition (altered prey distribution) and (iii) wolf spatial responses (altered wolf distribution independent of prey), we modeled the relationships between disturbances and indices of caribou-moose and caribou-wolf co-occurrence and then examined predation risk for caribou as a function of co-occurrence. We did not detect any relationships between anthropogenic disturbances, moose density, and caribou survival. Although caribou-moose co-occurrence increased predation risk, we observed both positive and negative relationships between disturbances and caribou-moose co-occurrence. In contrast, caribou-wolf co-occurrence increased predation risk and was positively correlated with anthropogenic linear features. Contrary to other boreal caribou populations, our analyses demonstrate stronger support for the direct effects of anthropogenic linear features on caribou-wolf spatial overlap, leading to greater risk for caribou. Our research highlights the need for region-specific management actions to conserve and recover widely distributed species.

A multiplex PCR test was evaluated to detect the DNA of three important dog haemoparasites by comparing with singular PCR counterpart on clinical blood samples of dogs in and around Chennai, Tamil Nadu, India. Initial screening of samples was done by microscopic examination of peripheral blood smear and singular PCR and those found exclusively positive for Babesia spp , Ehrlichia canis and Trypanosoma evansi and concurrent infections were used to standardize multiplex PCR. Amplicons of 619 bp, 377 bp and 227 bp corresponding to Babesia spp ( 18S rRNA gene), E. canis (VirB9 gene), and T.evansi (VSG gene ) respectively were amplified, without any non-specific amplification. The laboratory sensitivity (91.7% to 100%) and specificity (100%) of the multiplex PCR were calculated using ‘true positive’ and ‘true negative’ dog blood samples obtained in the initial screening process. Clinical blood samples from 287 dogs were screened using singular PCR and multiplex PCR tests for the presence of genome of Babesia spp, E. canis and T. evansi . The multiplex PCR was found to have high level of diagnostic specificity (97.5%–100%) in the detection of all three dog blood parasites and high level of diagnostic sensitivity (95%) in the detection of T. evansi from field level clinical blood samples compared to the singular PCR. However, the diagnostic sensitivity of the multiplex PCR was found to be low to moderate (40.45%–66.7%) in detection of Babesia spp and E. canis from field level clinical blood samples. The strength of agreement between singular and multiplex PCR assays was ‘moderate’ (0.445), ‘good’ (0.708) and ‘very good’ (0.968) in detection of DNA of Babesia spp, E. canis and T. evansi . The multiplex PCR was found to be 10 fold less sensitive in comparison with the singular PCR counterpart.

After more than two decades of implementation of the Habitats Directive (Directive 92/43/EEC), some fundamental aspects of the directive are still unclear, and subject to interpretive uncertainty, which limit its correct implementation. For example, obligations for Member States in situations where a protected population has almost, or has just, gone extinct are unclear. The isolated and protected population of wolves (Canis lupus) in the Sierra Morena region in Spain – the only wolf population in the southern half of the Iberian Peninsula – has been steadily declining to the point where it is doubtful whether any wolves are left. Using this illustrative example, we provide clarifications on the obligations by Member States in situations where populations are on the verge of extinction. Our analysis shows that Articles 6 and 12 of the Habitats Directive require Member States to restore populations that are quasi extinct. From a legal perspective, even the complete extinction of the species would not exonerate Member States from its obligations regarding the species in the Natura 2000 sites concerned. In this line, we argue that the Spanish authorities should not wait with recolonization, reinforcement and/or reintroduction actions until the complete absence of wolves in the Sierra Morena is conclusively proven. Two scenarios appear to meet legal requirements: i) active reinforcement/reintroduction, or ii) an active and effective policy towards a rapid natural recolonization of Sierra Morena by northern wolves. However, based on the observed wolf trends in Spain and Portugal during the past five decades, a reconnection between northern and Sierra Morena wolves seems unlikely in the foreseeable future even if actively promoted. Considering the urgency of actions required to avoid that this population will be the first wolf population to become extinct in Europe in modern times, in order to comply with European obligations, the adopting and carrying out a reintroduction/reinforcement scheme to restore the Sierra Morena wolf population is required. Such a scheme needs to be accompanied by a comprehensive enforcement plan to assure that reintroduced wolves will thrive.

This cross-sectional, observational, and descriptive study aims to investigate the epidemiology of Ehrlichia canis in healthy owned dogs from the Southeastern region of Rio de Janeiro, Brazil. Blood samples were collected from 390 households dogs. During the visits, an epidemiological questionnaire was filled out concerning the dogs’ characteristics as well as the environments in which they lived. The variables were analyzed using a bivariate test, while the correlation analysis between the variables was performed via a phi test. The variables that had p-values lower than 0.2 in the bivariate analysis and had a low or moderate correlation were selected for the multivariate analysis. The model that had the lowest Akaike information criterion (AIC) value was retained. Among the 390 blood samples tested, 24.8% were considered positive for E. canis. The parsimonious logistic regression model presented an AIC value of 408.75 and showed three variables that favored the presence of E. canis DNA in the tested dogs: the animal’s access to urban streets and neighborhoods (odds ratio [OR] = 1.91; p-value = 0.02; confidence interval [CI]: 1.14 − 3.18), tick infestation (OR = 2.01; p-value = 0.006; CI: 1.22 − 3.32), and poor hygienic conditions (OR = 2.19; p-value = 0.002; CI: 1.31 − 3.67). The model was considered well-calibrated based on the Hosmer–Lemeshow test (p = 0.39). According to the present study, dogs that have access to the street and neighborhood, are infested with ticks, and live under poor hygienic conditions are more likely to be infected with E. canis in the state of Rio de Janeiro, Brazil.

Dental microwear texture analysis (DMTA) has been instrumental in reconstructing dietary ecology of extinct and extant carnivorans. Current sampling methods for canids focus on lower second molars (m2), where the grinding of flesh and bone captures dental microwear indicative of diet. However, dental microwear on other biomechanically analogous grinding facets (i.e. the talonid basin on the lower carnassial, m1) might be comparable and could help dramatically increase sample sizes of fossil specimens, as carnassials are more frequently recovered and identifiable than lower m2s. Here, we quantify the degree to which dental microwear textures between grinding facets on lower first and second molars are similar in two species of extant canids, coyotes (Canis latrans) and gray wolves (Canis lupus). Casts of paired m1s and m2s for each individual were sampled from museum collections and analyzed for three microwear parameters that correlate with diet in carnivorans: anisotropy, complexity, and textural fill volume. Within wolves, the m1 talonid and m2 are indistinguishable in all DMTA parameters. In coyotes, grinding facets of the m1 talonid and m2 are indistinguishable in complexity and textural fill volume, but anisotropy values of m1s are significantly lower than those of m2s. Differences in anisotropy between species were unlikely to be driven by biomechanical shifts in bite force between the m1 talonid and m2, but could stem from a combination of subtle morphological variation and intra-tooth variation. Overall, these data suggest that regions across molars with similar functions yield similar dental microwear textures. Finally, to demonstrate the effect of increased sample size, we show how the combination of DMTA data from lower m1 talonids and lower m2s of the hypocarnivorous Phlaocyonini canids from the John Day Formation of eastern Oregon, USA, alters the size and shape, but not position, of their reconstructed dietary “niche” space and hence interpretations about dietary behavior.

Angiostrongylus vasorum is a nematode parasite of the pulmonary arteries and heart that infects domestic and wild canids. Dogs (Canis familiaris) and red foxes (Vulpes vulpes) are the most commonly affected definitive hosts. Recent studies suggest that angiostrongylosis is an emerging disease, and that red foxes may play an important role in the epidemiology of the parasite. Genetic analyses of parasites collected from dogs and foxes throughout Europe have shown that the same parasite haplotypes are commonly shared between different host species. However, the extent of genetic diversity within local A. vasorum populations and individual hosts is unknown. The objective of the present study was to assess the occurrence of genetic diversity among A. vasorum (a) recovered from different foxes within the Greater London area (a localised population, single worm per fox dataset); and (b) hosted within single foxes (multiple worms per fox dataset). During 2016, A. vasorum worms were collected from foxes culled for other purposes in London. DNA was extracted from each parasite and a partial fragment of the mitochondrial cytochrome oxidase subunit 1 (mtCOI) gene was amplified and sequenced. Sequences from the single worm dataset were compared with those published elsewhere. Combined, 19 haplotypes were described of which 15 were identified from foxes found in London, indicating that considerable genetic diversity can be detected within a local geographic area. Analysis of the multiple worm dataset identified 22 haplotypes defining worms recovered from just six foxes, emphasising the relevance of wild canines as reservoirs of genetic diversity. This is the first study to explore the genetic complexity of individual fox-hosted A. vasorum populations.

More than hundred urine samples, vaginal secretions, vulval and anal imprints as well as anal sac secretions, collected during various phases of the ovarian cycle, were evaluated with the HS-SPME/GC-MS technique. The results indicate there were differences among samples of urine collected during particular phases of the ovarian cycle. Unequivocally, the amount of carbonyl aromatic compounds, such as acetophenone (hypnone) and benzaldehyde, increased during the period of proestrus and estrus of the estrous cycle. This was accompanied by increases in methylketones, e.g., 2-octanone, 2-pentanone and 3-hexanone. Simultaneously, amounts of sulfide compounds (1-methylotiopropane; 1-methylotiobutane, 1-methylotipentane and dimethyl trisulfide) decreased during the period of estrus and abruptly increased in diestrus. These observations suggest a possible dual mechanism of interaction between males and females during and subsequent to the mating period, including the existence of both attracting and repelling signals. No significant changes were detected in samples other than urine. Further studies including a proteomic approach as well as behavioral assessments are suggested to identify the complete range of estrual semiochemical signals and to verify the semiochemical activities of identified substances.

Ehrlichia canis is the causative agent of canine monocytic ehrlichiosis (CME). While there is a high prevalence of CME in Thailand, genetic diversity of E. canis is still poorly defined. This study examined the molecular characteristics of E. canis using PCR and phylogenetic analysis of the dsb, gp19 and gp36 genes. DNA was extracted from 220 whole blood samples of naturally infected dogs, and all had clinical signs compatible with tick-borne diseases. Of these, 16.4% (36/220) provided positive E. canis DNA via the dsb and gp19 genes. However, only 13 out of the 36 samples (36.1%) were positive for the gp36 gene. Sequences of the dsb gene had very high identity (99–100%) with previously deposited E. canis sequences. Sequences of the gp19 gene were similar to those from US and Taiwanese genogroups (98.8–99.5% identity). Elucidation of genetic characteristics of E. canis based on the gp36 gene displayed 91.4–99.1% shared identity. There were 426–429 bp of a 5′ end pre-repeat tandem region, a 27 bp repetition with variable numbers of a tandem repeat (TR) region of 9 amino acid sequences (TEDSVSAPA), and a variable 3′ end region with sequence length depending on the isolate (72–93 bp). Phylogenetic trees of E. canis, particularly using the gp36 amino acid sequences, showed that the Thai strains fell into two phylogenetic clades contained within other worldwide E. canis strains. Alignment and phylogenetic analysis suggested that E. canis strains from Thailand could be divided into two genogroups, the US and Taiwanese genogroups. This study provides the first characterization of the dsb and gp19 genes of E. canis in Thailand, the results support the conclusion that the gp36 is a potential target for genotyping and elucidation of phylogenetic relationships among E. canis strains.

Aim Predation on livestock is one of the primary concerns for Mexican wolf (Canis lupus baileyi) recovery because it causes economic losses and negative attitudes toward wolves. Our objectives were to develop a spatial risk model of cattle depredation by Mexican wolves in the USA portion of their recovery area to help reduce the potential for future depredations. Location Arizona and New Mexico, USA. Methods We used a presence-only maximum entropy modeling approach (Maxent) to develop a risk model based on confirmed depredation incidents on public lands. In addition to landscape and human variables, we developed a model for annual livestock density using linear regression analysis of Animal Unit Month (AUM), and models for abundance of elk (Cervus canadensis), mule deer (Odocoileus hemionus) and white-tailed deer (Odocoileus virginiana) using Maxent, to include them as biotic variables in the risk model. We followed current recommendations for controlling model complexity and other sources of bias. Results The primary factors associated with increased risk of depredation by Mexican wolf were higher canopy cover variation and higher relative abundance of elk. Additional factors with increased risk but smaller effect were gentle and open terrain, and greater distances from roads and developed areas. Main conclusions The risk map revealed areas with relatively high potential for cattle depredations that can inform future expansion of Mexican wolf distribution (e.g., by avoiding hotspots) and prioritize areas for depredation risk mitigation including the implementation of active non-lethal methods in depredation hotspots. We suggest that livestock be better protected in or moved from potential hotspots, especially during periods when they are vulnerable to depredation (e.g. calving season). Our approach to create natural prey and livestock abundance variables can facilitate the process of spatial risk modeling when limitations in availability of abundance data are a challenge, especially in large-scale studies.

Nonlethal tools for reducing livestock depredations, such as livestock guardian dogs (LGDs; Canis familiaris), reduce lethal management of livestock predators and have been widely adopted by domestic sheep (Ovis aries) producers in the United States. However, compared with their success in reducing coyote (Canis latrans) depredations, commonly used LGD breeds appear less effective against wolves (Canis lupus). With more than 30 distinct LGD breeds found throughout the world, certain breeds may be more effective at deterring specific threats. We compared LGD breeds commonly used in the United States, collectively called whitedogs, with three European breeds selected for boldness toward carnivores, history of use in areas with wolves, lack of aggression toward humans, and size. We collected data on LGD behavior with sheep herds in Idaho, Montana, Oregon, Washington, and Wyoming in 2015 and 2016. We also developed a test to examine LGDs’ response to a simulated encounter with a wolf while on summer grazing range. Results from generalized linear mixed models of proportion of time spent in a given behavior indicate that few significant behavioral differences exist among tested breeds. Kangals tended to be more investigative when engaging a decoy, karakachans more vigilant, and transmontanos more able to decipher a threatening from unthreatening stimulus. Transmontanos also spent less time scanning than whitedogs, and there was a marginally significant effect of karakachans moving more than whitedogs. While these subtle behavioral differences may help livestock producers make tailored decisions in choosing the appropriate LGD for their needs and circumstance, our results suggest that behavioral differences among breeds may be less common than often suggested.

Canis mosbachensis is a widely distributed species from the Early to Middle Pleistocene in Europe. In Eastern Asia, Canis variabilis was supposed to be a local subspecies of C. mosbachensis, but such hypothesis has never been tested in details. Here we restudied the Canis materials from type locality, Loc.1 of ZKD (Zhoukoudian) of Middle Pleistocene, as well as other localities that produced important materials of this taxon (e.g. Loc.13 of ZKD). To better evaluate the variability of the Pleistocene Canis, a series of morphotypes are described. Based on morphotype and morphometric, the present study reveals there is only one species present at Loc.1 and Loc.13 of ZKD, which is quite similar to European C. mosbachensis and should be revised as a subspecies of C. mosbachensis, namely C. mosbachensis variabilis. C. m. variabilis is more derived and less hypercarnivorous than the Early Pleistocene Canis chihliensis (as well as European Canis etruscus and Canis arnensis) in craniodental characters, whereas it is less derived and less hypercarnivorous than Canis lupus. A review of fossil records of C. m. variabilis in China was made based on these different characters. Canids from Gongwangling, Lantian of Shaanxi Province (middle or late Early Pleistocene) are the earliest known representatives of C. mosbachensis in China, which are still more primitive than C. m. variabilis from the Middle Pleistocene sites of ZKD. Reliable records of C. mosbachensis are only known from the Early Pleistocene to the late Middle Pleistocene in Northern China. C. m. variabilis is not direct ancestor to C. lupus but a close relative of the latter, while it is far from the Early Pleistocene C. chihliensis. The latter remains more primitive characters that are more similar to those of European Canis etruscus and Canis arnensis.

The golden jackal (Canis aureus) is a medium-sized canid species native to Europe. This species is characterized by rapid large-scale expansion. A similar trend is also observed in Serbia, where the species is now distributed in more than a half of the territory. Although jackals prefer habitats in human-dominated landscapes, these animals have not been studied well enough from an eco-epidemiological point of view, and little is known about their potential for carrying zoonotic pathogens. In a study conducted during a three-year period (01/2010-02/2013), a total of 216 hunted or road-killed golden jackals were collected from 10 localities in Serbia. Ticks, when present, were removed, and after necropsy, spleen samples were collected from each animal. All tick and spleen samples were tested for the DNA of bacterial and protozoan tick-borne pathogens (Borrelia species, Bartonella species, Rickettsia species, Anaplasma species, Coxiella burnetii, Francisella species and Babesia species) by multiplex real-time PCR, conventional PCR and sequencing analyses. The DNA of Babesia canis was detected in nine out of 216 (4.2%) spleen samples, and two samples (0.9%) tested positive for Anaplasma phagocytophilum. In 118 ticks collected from jackals, the DNA of two Babesia species (Ba. canis and Ba. microti), three Borrelia species (Bo. garinii, Bo. valaisiana, and Bo. lusitaniae) and A. marginale was detected. From the aspect of public health surveillance, the potential role of the golden jackal in the maintenance of vector-borne zoonotic pathogens in Serbia must be considered, and further eco-epidemiological studies should be performed to determine the precise role of this animal species in zoonotic disease transmission cycles.

Brucella canis is a facultative intracellular pathogen responsible for canine brucellosis, a zoonotic disease that affects canines, causing abortions and reproductive failure; and the production of non-specific symptoms in humans. In 2005 the presence of B. canis in Antioquia was demonstrated and the strains were identified as type 2. The sequencing of the genome of a field strain denoted Brucella canis str. Oliveri, showed species-specific indel events, which led us to investigate the genomic characteristics of the B. canis strain isolated and to establish the phylogenetic relationships and the divergence time of B. canis str. Oliveri. Conventional PCR sequencing was performed in 30 field strains identifying 5 indel events recognized in B. canis str. Oliveri. ADN from Brucella suis, Brucella melitensis and vaccine strains from Brucella abortus were used as control, and it was determined that all of the studied field strains shared 4 out of the 5 indels of the sequenced Oliveri strain, indicating the presence of more than one strain circulating in the region. Phylogenetic analysis was performed with 24 strains of Brucella using concatenated sequences of genetic markers for species differentiation. The molecular clock hypothesis and Tajima's relative rate test were tested, showing that the Oliveri strain, similarly to other canis species, diverged from B. suis. The molecular clock hypothesis between Brucella species was rejected and an evolution rate and a similar genetic distance between the B. canis were demonstrated.

The aim of this study was to evaluate the relationship between naturally occurring Leishmania spp. infections in dogs (Canis familiaris) and the practical implications of the use of serological and molecular methods to confirm diagnoses. The study population consisted of 96 domestic dogs in southeastern Brazil. Serum samples were tested for the presence of anti-Leishmania immunoglobulin G (IgG) antibodies using four commercial canine visceral leishmaniasis kits. Dogs confirmed positive by immunofluorescence antibody test (IFAT) were culled and samples from mesenteric lymph nodes, spleen border, bone marrow and ear skin were taken and submitted to DNA extraction. PCR reactions were performed using primers that amplify a 300–350 bp fragment of the Leishmania ribosomal internal transcribed spacer 1 (ITS1) region. The ITS1 amplified products were analyzed by PCR-RFLP using Hae III restriction endonuclease. To confirm the Leishmania species detected by PCR, each purified sample was sequenced in duplicate. Of the 96 serum samples submitted to serological assays, 8 (8.3%) tested positive for Leishmania by IFAT, 4 (4.1%) by ELISA, 2 (2.1%) by rK39 RDT and 7 (7.3%) by DPP. Four of these infected dogs (50%) were found to be infected only by Leishmania braziliensis or Leishmania amazonensis, and their serum samples tested positive by IFAT and DPP. These findings demonstrate for the first time that cross-reactivity of L. braziliensis and L. amazonensis infection in dogs can be found using the DPP serum test. This is the first record of Leishmania (Leishmania) amazonensis confirmed by a specific molecular marker in dogs (Canis familiaris) from Belo Horizonte, Brazil.

In Brazil, active infections of Rickettsia spp. is confirmed in all regions, involving various species of ticks. During investigation of a new focus of Spotted Fever (SF) incidence in the state of Pernambuco, northeastern Brazil, tick species Rhipicephalus sanguineus sensu lato (Latreille), and Amblyomma pseudoconcolor were collected from one Canis familiaris and four Euphractus sexcinctus, respectively, and analized for the presence of rickettsial genes. Ten A. pseudoconcolor ticks (90.9%) were found to be infected with Rickettsia, whereas no evidence of Rickettsia spp. was found in R. sanguineus s. l. Genetic analysis based of five rickettsial genes showed that the detected strain is most closely related to Rickettsia amblyommatis (formerly Candidatus Rickettsia amblyommii). R. amblyommatis was, for the first time, detected in Amblyomma pseudoconcolor and the results pointed to this tick like a potential vector in the enzootic cycle of R. amblyommatis in a typical semiarid Brazilian savannah region. In conclusion, despite the need for further studies to confirm if R. amblyommatis was responsible for the observed case in the state of Pernambuco, the presence of this bacterium during an SF focussed investigation should be a major concern in terms of public health due the capacity of SF for rapid and extensive dispersion within Brazilian territory.

Leishmaniosis is, to date, considered the second most important emerging vector-borne protozoal disease in the world after malaria. The form of zoonotic visceral leishmaniosis found in the Mediterranean basin is caused by Leishmania infantum, and its life cycle includes the domestic dog and a phlebotomine sandfly vector. This complex epidemiological cycle and its high prevalence of subclinical infection, hinder the surveillance and control of L. infantum, and allows it to go unnoticed at the geographical endemicity limits of the parasite or in recently colonized areas. We, therefore, tested 102 wolves (Canis lupus) and 47 other wild carnivores in order to detect Leishmania DNA by means of PCR. Samples were collected from 2008 to 2014 in Asturias (northern Spain), a region considered non-endemic for the parasite. The results obtained provided valuable information regarding the prevalence of Leishmania in wild carnivores in Asturias and its geographic distribution in the region: an average prevalence of 33% for wolves and an overall prevalence of 40% for all the wild carnivores studied were reported, with a widespread presence of the parasite in the region and an apparent increase in its prevalence in wolves during the last decade. This suggests the usefulness of the wolf as a sentinel species for the detection and study of Leishmania in the field and confirms the value of wildlife sanitary surveillance programs for the detection and monitoring of hitherto disregarded diseases that affect domestic animals and humans., This is a contribution to the agreement between CSIC and Principado de Asturias.

Tick-borne diseases (TBDs), including emerging and re-emerging infectious diseases, are important threats to human and animal health worldwide. Indeed, the number of reported human and animal infectious cases of novel TBD agents has increased in recent decades. However, TBDs tend to be neglected, especially in resource-limited countries that often have limited diagnostic capacity. The aim of this molecular survey was to detect and characterise tick-borne pathogens (Babesia, Theileria, and Hepatozoon parasites and Anaplasmataceae bacteria) in domestic dogs in Zambia. In total, 247 canine peripheral blood samples were collected in Lusaka, Mazabuka, Monze, and Shangombo. Conventional PCR to detect the selected pathogens was performed using DNA extracted from canine blood. One hundred eleven samples were positive for protozoa and 5 were positive for Anaplasmataceae. Sequencing of thirty-five randomly selected protozoa-positive samples revealed the presence of Babesia rossi, Babesia vogeli, and Hepatozoon canis 18S rDNA. Based on these sequences, a multiplex PCR system was developed to yield PCR products with different amplicons, the size of which depended on the parasite species; thus, each species could be identified without the need for sequence analysis. Approximately 40% of dogs were positive for H. canis. In particular, the positive rate (75.2%) of H. canis infection was significantly higher in Shangombo than in other sampling sites. Multiplex PCR assay detected B. rossi and B. vogeli infections in five and seven dogs, respectively, indicating that this approach is useful for detecting parasites with low prevalence. Sequencing analysis of gltA and groEL genes of Anaplasmataceae revealed that two and one dogs in Lusaka were infected with Anaplasma platys and Ehrlichia canis, respectively. The data indicated that Zambian dogs were infected with multiple tick-borne pathogens such as H. canis, B. rossi, B. vogeli, A. platys, E. canis and uncharacterized Ehrlichia sp. Since some of these parasites are zoonotic, concerted efforts are needed to raise awareness of, and control, these tick-borne pathogens.

Large carnivores have historically been decreasing worldwide, often as a result of human-carnivore conflicts. However, large carnivores are recovering throughout Europe, and European management scenarios can provide important insights into broad issues related to human-large carnivore existence. After becoming almost extinct in Sweden during the mid-19th century the Swedish grey wolf (Canis lupus) population has now recovered. Current national wolf management aims to promote distribution shifts from the current areas in central Sweden, potentially also into a previously exempt reindeer husbandry area. Prior wolf re-introductions have highlighted the necessity of pro-active management for colonization success. Identification of likely range expansion areas could therefore be paramount for a successful Swedish wolf management. We characterized the demographic and spatial progression of Swedish wolves during 2001–2015 and used a MaxEnt approach to species distribution models to identify potential range expansion areas. The Swedish wolf population had expanded from 10 to almost 60 reproductions or territorial pairs, and increased in both range size and density. Our distribution models suggested that Swedish wolf management may face trade-offs between costs of hosting wolves in densely populated areas in southern Sweden with cattle and sheep and the costs of allowing wolves to expand into reindeer husbandry areas with associated cultural and economic consequences. Spatially explicit data on the economic, social and cultural factors associated with wolf conflict and acceptance may be paramount for developing optimal management strategies in the face of such a trade-off.

The Mexican wolf (Canis lupus baileyi) was included in the 1973 Endangered Species Act listing of the gray wolf (C. lupus), but then listed separately as a subspecies in 2015. Early accounts of its range included the Sierra Madre Occidental of Mexico, southeastern Arizona, southwestern New Mexico, and sometimes western Texas, supported by ecological, biogeographic, and morphological data. There have been multiple unsuccessful attempts to revise the original 1982 recovery plan and identify areas suitable for Mexican wolf reintroduction. Despite the fact that 90% of its historical range is in Mexico and widespread suitable habitat exists there, previous draft recovery plans recommended recovery mostly outside of Mexico and well north of the subspecies' historical range. Planning recovery outside historical range of this subspecies is fraught with problems that may compromise, thwart, or impede successful recovery. Dispersal of Mexican wolves northward and continued movements southward by Northwestern wolves (C. l. occidentalis), along with allowing establishment of Mexican wolves north of their historical range before they are recovered, may lead to premature and detrimental intraspecific hybridization. Interbreeding of Northwestern wolves from Canadian sources and Mexican wolves does not represent the historical cline of body size and genetic diversity in the Southwest. If Northwestern wolves come to occupy Mexican wolf recovery areas, these physically larger wolves are likely to dominate smaller Mexican wolves and quickly occupy breeding positions, as will their hybrid offspring. Hybrid population(s) thus derived will not contribute towards recovery because they will significantly threaten integrity of the listed entity. Directing Mexican wolf recovery northward outside historical range threatens the genetic integrity and recovery of the subspecies, is inconsistent with the current 10(j) regulations under the ESA, is unnecessary because large tracts of suitable habitat exist within historical range, is inconsistent with the concepts of restoration ecology, and disregards unique characteristics for which the Mexican wolf remains listed.

Skulls from 392 grey wolves (Canis lupus) were examined macroscopically according to predefined criteria. Two hundred and seven skulls were included in this study, comprised of 124 young adults (59.9%) and 83 adults (40.1%); of these, 65 (31.4%) specimens were from male wolves and 104 (50.3%) were from females, with 38 (18.4%) of unknown sex. Out of 8,694 possible teeth, 8,339 (95.9%) were present for evaluation. Fifty-five teeth (15.5%) were absent congenitally, 30 (8.5%) were lost during life and 270 (76.1%) were lost artefactually post mortem. Skeletal or dental malocclusion was present in 37 specimens (17.9%), with level bite being the most commonly encountered malocclusion. Enamel hypoplasia was present in five skulls (2.4%), affecting eight teeth (0.1%) in total. An abnormal number of roots was found on 23 teeth (0.3%) on 13 skulls (6.3%). Persistent deciduous teeth occurred in two (1.0%) specimens, affecting one (0.01%) tooth each. Fenestration or dehiscence was found associated with 203 teeth (2.4%) in 72 skulls (34.8%). Periodontitis was noted on 115 skulls (55.6%)and 1,000 teeth (11.5%), affecting significantly more adults (n=63, 75.0%) than young adults (n=52, 41.9%; P

Domestic dogs (Canis familiaris) are social animals and, therefore, social interactions with conspecifics are crucial for their welfare. However, in kennelled dogs, the ability to interact with conspecifics may be limited. Swiss military dogs, for instance, are kept individually without direct contact to conspecifics. Here we asked whether short-term exposure to conspecifics may be beneficial for dogs kept in isolation. The treatment lasted for eight weeks and consisted of one session weekly of three hours of social exposure. During social exposure, focal dogs were allowed direct contact with conspecifics but were supervised by the experimenter who intervened when necessary to prevent dogs from attacking one another. Immediately before and after the treatment phase, the dogs’ reactions towards unfamiliar objects (including a dog model) and an unfamiliar male dog (stimulus dog) were assessed both in experimental dogs (with social exposure, n = 29) and control dogs (without social exposure, n = 27). We predicted a positive effect of the social exposure, i.e. less offensive and defensive behaviours shown towards both the unfamiliar dog model and stimulus dog. In accordance with our predictions, experimental dogs showed a greater decrease in offensive and defensive behaviours compared to control dogs. Although none of these dogs had been socialised conventionally like family dogs are, we found a clear positive effect of social exposure in adult dogs on their social behaviour. Thus, working dogs, which are otherwise kept singly, may benefit from temporary social exposure in terms of both their working ability and their wellbeing.

The objectives of the present study were to investigate the presence of the Babesia parasites in Dermacentor reticulatus ticks along its current distribution range in Lithuania and Latvia, and Ixodes ricinus in Lithuania, to characterize the detected Babesia spp. using partial sequencing of 18S rRNR gene, and to determine the prevalence of the Babesia pathogens in ticks from different locations of Lithuania and Latvia. From 2013 to 2015, four D. reticulatus nymphs and 2255 questing adults were collected from 40 locations in Lithuania, and 181 questing adult D. reticulatus specimens from 11 locations in Latvia. In Lithuania, 16 questing I. ricinus nymphs and 354 questing adults were collected from 12 locations with the sympatric existence of D. reticulatus and I. ricinus. In Lithuania, Babesia spp. were detected in 1.2% (26/2259) of D. reticulatus and in 9.5% (35/370) of I. ricinus ticks. The overall prevalence of Babesia in D. reticulatus ticks from Latvia was 2.8% (5/181). Sequence analysis of partial 18S rRNA gene of positive samples indicated the presence of B. canis and B. venatorum in D. reticulatus ticks, whereas B. microti and B. venatorum were detected in I. ricinus ticks. Our study is the first investigation on the prevalence and molecular characterization of zoonotic B. canis, B. venatorum, and B. microti in ixodid ticks in Lithuania, and of B. canis in D. reticulatus ticks in Latvia. It is also the first report of B. venatorum in D. reticulatus ticks.

Canine monocytic ehrlichiosis (CME), caused by a rickettsial bacterium, Ehrlichia canis, is distributed worldwide, particularly in tropical and subtropical regions. Transmission of E. canis is primarily mediated by the vector tick, Rhipicephalus sanguineus sensu lato and the bacteria then infect and replicate in monocytes and macrophages. Many cases are seen in veterinary hospitals and treated routinely; however, the genetic variation of E. canis strains found in the Philippines has been poorly investigated to date. In this study, the 16S rRNA gene and the gp200 gene of E. canis were detected by polymerase chain reaction from an infected dog in the Philippines, and the deduced amino acid sequence of the gp200 gene was subjected to a phylogenetic analysis. The Philippine genotype formed a cluster with the Taiwan genotype, and was somewhat divergent from the USA and Brazil strains. This suggested that E. canis underwent evolution in East and Southeast Asia, confirming the utility of the gp200 gene for the assessment of genetic relationships among strains.