Molecular Detection and Genetic Variability of Hepatozoon canis in Golden Jackals (Canis aureus L. 1758) in Serbia.

Hepatozoon canis is a protozoan tick-borne parasite infecting domestic and wild canids, including foxes, wolves, and jackals. It is mainly found in dogs but has also been detected in several wild carnivores, including foxes, wolves, and jackals. Host transmission primarily occurs through the ingestion of infected ticks, typically Rhipicephalus sanguineus, with documented instances of transplacental transmission from infected females to cubs. In Serbia, the golden jackal is common throughout the country, and its population has increased in recent years. Previous research has documented the presence of several vector-borne pathogens in the jackal population in Serbia, so we conducted this study to determine the presence, prevalence, and genetic variability of H. canis. Over eleven years (2010-2020), 114 animal samples were collected from 23 localities in Serbia. A total of 90/114 (78.95%) jackals were positive for H. canis, and they came from 22 localities. Among 15 juveniles, almost half (6/15 (40%)) tested positive for H. canis. In addition to the high prevalence, high genetic variability of the pathogen was also found. According to the mutated positions, four sequence types (S4-S7) of H. canis were determined. Based on our earlier research on the grey wolf and on this study, it can be observed that various sequence types of H. canis circulate within wild canid populations in Serbia. The prevalence of H. canis infection in wild carnivores raises significant concerns for wildlife conservation and animal health. Infected animals may act as reservoirs for the disease, posing a potential risk to domestic animals by acting as a source of infection.

Mammal responses to global changes in human activity vary by trophic group and landscape.

Wildlife must adapt to human presence to survive in the Anthropocene, so it is critical to understand species responses to humans in different contexts. We used camera trapping as a lens to view mammal responses to changes in human activity during the COVID-19 pandemic. Across 163 species sampled in 102 projects around the world, changes in the amount and timing of animal activity varied widely. Under higher human activity, mammals were less active in undeveloped areas but unexpectedly more active in developed areas while exhibiting greater nocturnality. Carnivores were most sensitive, showing the strongest decreases in activity and greatest increases in nocturnality. Wildlife managers must consider how habituation and uneven sensitivity across species may cause fundamental differences in human-wildlife interactions along gradients of human influence.

Rehabilitation and release of orphaned Eurasian lynx (Lynx lynx) in Europe: Implications for management and conservation.

Rehabilitation of injured or immature individuals has become an increasingly used conservation and management tool. However, scientific evaluation of rehabilitations is rare, raising concern about post-release welfare as well as the cost-effectiveness of spending scarce financial resources. Over the past 20 years, events of juvenile Eurasian lynx presumably orphaned have been observed in many European lynx populations. To guide the management of orphaned lynx, we documented survival, rehabilitation and fate after the release and evaluated the potential relevance of lynx orphan rehabilitation for population management and conservation implications. Data on 320 orphaned lynx was collected from 1975 to 2022 from 13 countries and nine populations. The majority of orphaned lynx (55%) were taken to rehabilitation centres or other enclosures. A total of 66 orphans were released back to nature. The portion of rehabilitated lynx who survived at least one year after release was 0.66. Release location was the best predictor for their survival. Of the 66 released lynx, ten have reproduced at least once (8 females and 2 males). Conservation implications of rehabilitation programmes include managing genetic diversity in small, isolated populations and reintroducing species to historical habitats. The lynx is a perfect model species as most reintroduced populations in Central Europe show significantly lower observed heterozygosity than most of the autochthonous populations, indicating that reintroduction bottlenecks, isolation and post-release management have long-term consequences on the genetic composition of populations. The release of translocated orphans could be a valuable contribution to Eurasian lynx conservation in Europe. It is recommended to release orphans at the distribution edge or in the frame of reintroduction projects instead of a release in the core area of a population where it is not necessary from a demographic and genetic point of view. Rehabilitation programmes can have conservation implications that extend far beyond individual welfare benefits.

The Occurrence and Diversity of Tick-Borne Pathogens in Small Mammals from Serbia.

Background: Despite abundance of small mammals in Serbia, there is no information on their role in the epidemiology of tick-borne diseases (TBDs). This retrospective study aimed to identify different tick-borne pathogens (TBPs) in small mammals in Serbia collected during 2011. Materials and Methods: A total of 179 small mammals were collected from seven different localities in Serbia. The five localities belong to the capital city of Serbia-Belgrade: recreational areas-Ada Ciganlija, Titov gaj, and Kosutnjak as well as mountainous suburban areas used for hiking-Avala and Kosmaj. The locality Veliko Gradiste is a tourist place in northeastern Serbia, whereas the locality Milosev Do is a remote area in western Serbia with minor human impact on the environment. Results: The results of the presented retrospective study are the first findings of Rickettsia helvetica, Rickettsia monacensis, Neoehrlichia mikurensis, Borrelia afzelii, Borrelia miyamotoi, Babesia microti, Hepatozoon canis, and Coxiella burnetii in small mammals in Serbia. The presence of R. helvetica was confirmed in two Apodemus flavicollis, the presence of one of the following pathogens, R. monacensis, B. afzelii, H. canis, Ba. microti, and N. mikurensis was confirmed in one A. flavicollis each, whereas the presence of B. miyamotoi was confirmed in one Apodemus agrarius. Coinfection with B. afzelii and Ba. microti was confirmed in one A. flavicollis. DNA of C. burnetii was detected in 3 of 18 pools. Conclusions: The results confirm that detected pathogens circulate in the sylvatic cycle in Serbia and point to small mammals as potential reservoir hosts for the detected TBPs. Further large-scale studies on contemporary samples are needed to clarify the exact role of particular small mammal species in the epidemiology of TBDs caused by the detected pathogens.

Behavioral responses of terrestrial mammals to COVID-19 lockdowns.

COVID-19 lockdowns in early 2020 reduced human mobility, providing an opportunity to disentangle its effects on animals from those of landscape modifications. Using GPS data, we compared movements and road avoidance of 2300 terrestrial mammals (43 species) during the lockdowns to the same period in 2019. Individual responses were variable with no change in average movements or road avoidance behavior, likely due to variable lockdown conditions. However, under strict lockdowns 10-day 95th percentile displacements increased by 73%, suggesting increased landscape permeability. Animals' 1-hour 95th percentile displacements declined by 12% and animals were 36% closer to roads in areas of high human footprint, indicating reduced avoidance during lockdowns. Overall, lockdowns rapidly altered some spatial behaviors, highlighting variable but substantial impacts of human mobility on wildlife worldwide.

Assessment of macro-, trace- and toxic elements in Small Indian mongoose, Herpestes auropunctatus (Hodgson, 1836), from Montenegro: potential use for biomonitoring.

The aim of this study was to determine the concentrations of 28 elements in the liver, kidney, muscle, and heart of a small Indian mongoose from Montenegro. Element concentrations were determined using inductively coupled plasma optical emission spectrometry (ICP-OES). Significant differences in elemental concentrations between analyzed tissues were observed for Ag, Al, Cu, Fe, Hg, K, Mg, Mn, Mo, Na, P, S, Se, and Zn. The concentrations of Ag, Fe, Mo, and S were significantly different between all analyzed tissues. Muscle tissue is characterised by higher concentrations of some macroelements (K, Mg, S) and liver tissue by higher concentrations of some trace elements (Fe, Mn, Mo, and Zn). Cardiac tissue could be the target organ for bioaccumulation of Ag and Se. The molar ratio Se:Hg was higher than 1 in all studied tissues. There were few significant differences between element concentrations in mongooses from three analyzed (Airport, Saline, Inland) groups. Significantly higher concentrations of Ag and Se were found in liver tissue of Inland mongooses compared to animals from Airport and Saline. There were few significant differences (Zn in liver and muscle, Fe in muscle) in element concentrations between females and males.

High Prevalence and Genetic Variability of Hepatozoon canis in Grey Wolf (Canis lupus L. 1758) Population in Serbia.

Wild canids are globally recognised as hosts and reservoirs of a large number of ecto- and endoparasites. Data that reveal the importance of the grey wolf (Canis lupus L.1758) in the spread of hepatozoonosis are very scarce. There are a large number of different potential host species that can be infected by Hepatozoon canis, but the most common are domestic and wild carnivores, such as dogs, jackals, foxes, and wolves. In this study, the epidemiological significance of the grey wolf as a host for the pathogen was analysed for the first time in Serbia, as well as the genetic variability of H. canis. The presence of H. canis in wolf spleens has been demonstrated using molecular methods. A total of 107 wolf spleen samples from 30 localities in Serbia were analysed. The presence of H. canis was confirmed in 62 (57.94%) individuals from 26 out of 30 localities. According to the analysis, the sampled H. canis sequences were found to be characterised by a certain heterogeneity. Based on five mutated nucleotide sites in the sequences, H. canis could be divided into five sequence types, S1 to S5. The five sequence types can potentially circulate in grey wolf populations as well as among other domestic and wild canids. This study is the first confirmation of the presence of H. canis in grey wolf populations in Serbia. Considering that the role of this vector-borne disease is poorly researched in wild carnivores, it is very important to indicate the role of this species in the circulation of this pathogen in natural ecosystems.

First Molecular Detection of Pneumocystis spp. in the Golden Jackal (Canis aureus).

Forty-six golden jackals (Canis aureus) were collected between November 2020 and February 2021 in five counties of Serbia. Lung samples were screened for the presence of Pneumocystis DNA by pan-Pneumocystis PCR on the mtLSU rRNA gene obtaining PCR products of 370 bp in length. Pneumocystis DNA was detected in the lungs from 6/46 (13.04%) golden jackals. Four were females and two were males; four were classified as adults and two as subadults. Positive samples were confirmed in 4/5 investigated counties. No gross pathologic lung lesions were observed in this study. The sequences of Pneumocystis spp. from golden jackals were identical to one another and showed the highest similarity with Pneumocystis spp. sequences of dogs (98% nucleotide identity). The genetic variation was comparable to Pneumocystis spp. of raccoon dogs (95-97% nucleotide identity), red foxes (91-95% nucleotide identity), ferrets (86% nucleotide identity), and another Pneumocystis type in dogs (P. canis Ck2, 81% nucleotide identity) was higher. Golden jackals may be carriers and may play a nonnegligible role in the spread of Pneumocystis spp. Although this finding cannot be directly related to any clinical manifestation or pathologic lesions, a possible role in the exacerbation of different pulmonary disorders should be considered.

First Report of Alveolar Hydatid Disease (Echinococcus multilocularis) in a Golden Jackal (Canis aureus).

BACKGROUND: Alveolar hydatid disease caused by the tapeworm Echinococcus multilocularis is a parasitic disease present in the northern hemisphere. Echinococcus multilocularis is a parasite of canid and felid carnivores as definitive hosts, and small mammals, particularly rodents as intermediate hosts. Other animal species and humans can be aberrant intermediate hosts for this parasite. It is known that besides acting as definitive hosts, domestic dogs can rarely become infected with the larval form of E. multilocularis and develop alveolar echinococcosis; however, a role of wild canids as aberrant intermediate hosts has not been documented until now. To the best of our knowledge the present paper provides the first description of alveolar hydatid disease in a golden jackal (Canis aureus). CASE PRESENTATION: Necropsy of the yearling female animal found a large, round, tumor-like mass, 20 cm in diameter, with a rough, multilobulated surface in the abdominal cavity, connected to the liver and omentum. On the cut surface this tumor-like lesion was multicystic, with a number of locular cavities filled with a clear yellowish to orange watery fluid and a large area of necrosis in the central part of the mass. Histopathology revealed multiple cystic spaces separated by fibrous sheaths and inflammatory cells-lymphocytes, plasma cells, neutrophil and eosinophil granulocytes. The cysts contained either pale, hyaline, eosinophilic laminar and occasionally amorphous, acellular, PAS-positive structures, or metacestodes with invaginated protoscolices. In several cysts round calcified bodies (calcareous corpuscles) were noted. Microscopic examination showed everted and inverted protoscolices which were attached to fragments of the brood capsule or free in hydatid fluid. By comparing consensus nucleotide sequence of 457 bp obtained by PCR reaction with sequences deposited in NCBI GenBank it is determined that it was 100% identical with E. multilocularis sequences under accession numbers MH259778.1, MH259776.1, AB668376.1, EU704124.1 and AB018440.2. CONCLUSIONS: The present paper provides a proof that the golden jackal, besides being a definitive host, can also serve as the aberrant intermediate host for E. multilocularis.

Cryptic Diversity of the European Blind Mole Rat Nannospalax leucodon Species Complex: Implications for Conservation.

We explored the cryptic speciation of the Nannospalax leucodon species complex, characterised by intense karyotype evolution and reduced phenotypic variability that has produced different lineages, out of which 25 are described as chromosomal forms (CFs), so many cryptic species remain unnoticed. Although some of them should be classified as threatened, they lack the official nomenclature necessary to be involved in conservation strategies. Reproductive isolation between seven CFs has previously been demonstrated. To investigate the amount and dynamics of genetic discrepancy that follows chromosomal changes, infer speciation levels, and obtain phylogenetic patterns, we analysed mitochondrial 16S rRNA and MT-CYTB nucleotide polymorphism among 17 CFs-the highest number studied so far. Phylogenetic trees delineated 11 CFs as separate clades. Evolutionary divergence values overlapped with acknowledged higher taxonomic categories, or sometimes exceeded them. The fact that CFs with higher 2n are evolutionary older corresponds to the fusion hypothesis of Nannospalax karyotype evolution. To participate in conservation strategies, N. leucodon classification should follow the biological species concept, and proposed cryptic species should be formally named, despite a lack of classical morphometric discrepancy. We draw attention towards the syrmiensis and montanosyrmiensis CFs, estimated to be endangered/critically endangered, and emphasise the need for detailed monitoring and population survey for other cryptic species.

Heartworm Disease in Jackals: Unusual Location of Dirofilaria immitis.

PURPOSE: The knowledge of heartworm disease in free ranging wild canid populations is limited. As it is very difficult to monitor this disease in live animals, sporadic findings are mostly obtained by examining culled individuals of game species. METHODS: As a part of a broader study on jackal (Canis aureus) ecology in Serbia, the necropsy of legally hunted animals was performed in 2020. RESULTS: Two cases of heartworm infection with unusually located adult heartworms were diagnosed. The first case is an adult female jackal harvested in the vicinity of the capital city of Belgrade. Three adult specimens of Dirofilaria immitis were found in the right atrium of the heart, and nine adult specimens were located at an atypical site, in the vena cava caudalis. Very similar, the second case was in an adult female jackal harvested in the vicinity of Backi Monostor. In this case, six adult specimens of D. immitis were found in the right atrium of the heart and six more adult specimens in the vena cava caudalis. CONCLUSION: Although this nematode generally resides in the pulmonary arteries and the right ventricle of the heart, rare findings in other large blood vessels can occur. To the best of our knowledge, these are the first and only two confirmed cases of adult D. immitis found in vena cava caudalis in jackals in Europe.

Next-generation phylogeography resolves post-glacial colonization patterns in a widespread carnivore, the red fox (Vulpes vulpes), in Europe.

Carnivores tend to exhibit a lack of (or less pronounced) genetic structure at continental scales in both a geographic and temporal sense and this can confound the identification of post-glacial colonization patterns in this group. In this study we used genome-wide data (using genotyping by sequencing [GBS]) to reconstruct the phylogeographic history of a widespread carnivore, the red fox (Vulpes vulpes), by investigating broad-scale patterns of genomic variation, differentiation and admixture amongst contemporary populations in Europe. Using 15,003 single nucleotide polymorphisms (SNPs) from 524 individuals allowed us to identify the importance of refugial regions for the red fox in terms of endemism (e.g., Iberia). In addition, we tested multiple post-glacial recolonization scenarios of previously glaciated regions during the Last Glacial Maximum using an Approximate Bayesian Computation (ABC) approach that were unresolved from previous studies. This allowed us to identify the role of admixture from multiple source population post-Younger Dryas in the case of Scandinavia and ancient land-bridges in the colonization of the British Isles. A natural colonization of Ireland was deemed more likely than an ancient human-mediated introduction as has previously been proposed and potentially points to a larger mammalian community on the island in the early post-glacial period. Using genome-wide data has allowed us to tease apart broad-scale patterns of structure and diversity in a widespread carnivore in Europe that was not evident from using more limited marker sets and provides a foundation for next-generation phylogeographic studies in other non-model species.

Seasonal and diel movement patterns of brown bears in a population in southeastern Europe.

Most animals concentrate their movement into certain hours of the day depending on drivers such as photoperiod, ambient temperature, inter- or intraspecific competition, and predation risk. The main activity periods of many mammal species, especially in human-dominated landscapes, are commonly set at dusk, dawn, and during nighttime hours. Large carnivores, such as brown bears, often display great flexibility in diel movement patterns throughout their range, and even within populations, striking between individual differences in movement have been demonstrated. Here, we evaluated how seasonality and reproductive class affected diel movement patterns of brown bears of the Dinaric-Pindos and Carpathian bear populations in Serbia. We analyzed the movement distances and general probability of movement of 13 brown bears (8 males and 5 females) equipped with GPS collars and monitored over 1-3 years. Our analyses revealed that movement distances and probability of bear movement differed between seasons (mating versus hyperphagia) and reproductive classes. Adult males, solitary females, and subadult males showed a crepuscular movement pattern. Compared with other reproductive classes, females with offspring were moving significantly less during crepuscular hours and during the night, particularly during the mating season, suggesting temporal niche partitioning among different reproductive classes. Adult males, solitary females, and in particular subadult males traveled greater hourly distances during the mating season in May-June than the hyperphagia in July-October. Subadult males significantly decreased their movement from the mating season to hyperphagia, whereas females with offspring exhibited an opposite pattern with almost doubling their movement from the mating to hyperphagia season. Our results provide insights into how seasonality and reproductive class drive intrapopulation differences in movement distances and probability of movement in a recovering, to date little studied, brown bear population in southeastern Europe.

Population genetic structure of European wildcats inhabiting the area between the Dinaric Alps and the Scardo-Pindic mountains.

Habitat fragmentation and loss have contributed significantly to the demographic decline of European wildcat populations and hybridization with domestic cats poses a threat to the loss of genetic purity of the species. In this study we used microsatellite markers to analyse genetic variation and structure of the wildcat populations from the area between the Dinaric Alps and the Scardo-Pindic mountains in Slovenia, Croatia, Serbia and North Macedonia. We also investigated hybridisation between populations of wildcats and domestic cats in the area. One hundred and thirteen samples from free-leaving European wildcats and thirty-two samples from domestic cats were analysed. Allelic richness across populations ranged from 3.61 to 3.98. The observed Ho values ranged between 0.57 and 0.71. The global FST value for the four populations was 0.080 (95% CI 0.056-0.109) and differed significantly from zero (P < 0.001). The highest FST value was observed between the populations North Macedonia and Slovenia and the lowest between Slovenia and Croatia. We also found a signal for the existence of isolation by distance between populations. Our results showed that wildcats are divided in two genetic clusters largely consistent with a geographic division into a genetically diverse northern group (Slovenia, Croatia) and genetically eroded south-eastern group (Serbia, N. Macedonia). Hybridisation rate between wildcats and domestic cats varied between 13% and 52% across the regions.

Positive selection on the MHC class II DLA-DQA1 gene in golden jackals (Canis aureus) from their recent expansion range in Europe and its effect on their body mass index.

BACKGROUND: In Europe, golden jackals (Canis aureus) have been expanding their range out of the southern and southeastern Balkans towards central Europe continually since the 1960s. Here, we investigated the level of functional diversity at the MHC class II DLA-DQA1 exon 2 in golden jackal populations from Bulgaria, Serbia, and Hungary. Specifically, we tested for positive selection on and geographic variation at that locus due to adaptation to supposedly regionally varying pathogenic landscapes. To test for potential fitness effects of different protein variants on individual body condition, we used linear modeling of individual body mass indexes (bmi) and accounted for possible age, sex, geographical, and climatic effects. The latter approach was performed, however, only on Serbian individuals with appropriate data. RESULTS: Only three different DLA-DQA1 alleles were detected, all coding for different amino-acid sequences. The neutrality tests revealed no significant but positive values; there was no signal of spatial structuring and no deviation from the Hardy-Weinberg equilibrium across the studied range of expansion. However, we found a signal of trans-species polymorphism and significant test results for positive selection on three codons. Our information-theory based linear modeling results indicated an effect of ambient temperature on the occurrence of individual DLA-DQA1 genotypes in individuals from across the studied expansion range, independent from geographical position. Our linear modeling results of individual bmi values indicated that yearlings homozygous for DLA-DQA1*03001 reached values typical for adults contrary to yearlings carrying other genotypes (protein combinations). This suggested better growth rates and thus a possible fitness advantage of yearlings homozygous for DLA-DQA1*03001. CONCLUSIONS: Our results indicate a demographic (stochastic) signal of reduced DLA-DQA1 exon 2 variation, in line with the documented historical demographic bottleneck. At the same time, however, allelic variation was also affected by positive selection and adaptation to varying ambient temperature, supposedly reflecting geographic variation in the pathogenic landscape. Moreover, an allele effect on body mass index values of yearlings suggested differential fitness associated with growth rates. Overall, a combination of a stochastic effect and positive selection has shaped and is still shaping the variation at the studied MHC locus.

Population genetic structure of wolves in the northwestern Dinaric-Balkan region.

The Balkan Peninsula and the Dinaric Mountains possess extraordinary biodiversity and support one of the largest and most diverse wolf (Canis lupus) populations in Europe. Results obtained with diverse genetic markers show west-east substructure, also seen in various other species, despite the absence of obvious barriers to movement. However, the spatial extent of the genetic clusters remains unresolved, and our aim was to combine fine-scale sampling with population and spatial genetic analyses to improve resolution of wolf genetic clusters. We analyzed 16 autosomal microsatellites from 255 wolves sampled in Slovenia, Croatia, Bosnia and Herzegovina (BIH), and Serbia and documented three genetic clusters. These comprised (1) Slovenia and the regions of Gorski kotar and Lika in Croatia, (2) the region of Dalmatia in southern Croatia and BIH, and (3) Serbia. When we mapped the clusters geographically, we observed west-east genetic structure across the study area, together with some specific structure in BIH-Dalmatia. We observed that cluster 1 had a smaller effective population size, consistent with earlier reports of population recovery since the 1980s. Our results provide foundation for future genomic studies that would further resolve the observed west-east population structure and its evolutionary history in wolves and other taxa in the region and identify focal areas for habitat conservation. They also have immediate importance for conservation planning for the wolves in one of the most important parts of the species' European range.

Detection and genotyping of Toxoplasma gondii in wild canids in Serbia.

To gain insight into the population structure of the protozoan parasite Toxoplasma gondii in wildlife in Serbia, multiplex nested PCR- RFLP was performed on T. gondii DNA positive samples of heart tissue of three wild canid species, including red foxes (Vulpes vulpes), golden jackals (Canis aureus) and grey wolves (Canis lupus). Out of 110 samples (28 red foxes, 72 golden jackals, 10 wolves), 30 (27.3%) were positive for T. gondii DNA, of which 24 were genotyped (nine red foxes, 13 golden jackals and two wolves). The results showed a large dominance of lineage II (n = 20, 83.4%) over lineage III (n = 4, 16.7%) strains. Lineage II comprised four archetypes, 14 variants and one which remained undetermined, while within lineage III there were three archetypes and one variant strain. However, a notable difference in diversity was observed among the examined canids, with eight (88.9%) of the nine strains detected in red foxes distinct, vs. six (46.1%) distinct strains of the 13 detected in golden jackals. Since anthropogenic food sources are a major reservoir of archetypal lineage II and III T. gondii strains, the results suggest that the golden jackal may be outcompeting the red fox, forcing it to hunt wild mammals and birds for sustenance, thereby increasing its chances of infection with variant strains. In addition to presenting the first data on the T. gondii population structure in wild canids in Serbia, this is the first report ever on T. gondii genotypes in golden jackals, a species with an increasing presence in Europe and another important reservoir of domestically circulating T. gondii strains.

Spatial Subchondral Bone Density Reflecting Joint Loading of the Talus in Different Canidae.

OBJECTIVE: Subchondral bone density distribution can be used to study joint biomechanics non-invasively. Differences in joint loading between related species can aid in the understanding of joint loading and the development of certain types of orthopaedic pathology. This study was conducted to evaluate density distribution in the subchondral bone of the talus of different Canidae species, as a parameter reflecting the long-term joint loading in the tarsocrural joint. MATERIALS AND METHODS: The tarsal joints of cadaveric dogs of different breeds were included, that is, German Shepherd (n = 5), Bouvier des Flandres (n = 3) and Labrador Retriever (n = 6).Additionally, golden jackals (n = 5) (Canis aureus) and wolves (n = 5) (Canis lupus) were included. Consecutive computed tomography slices were made and the subchondral bone density distribution was evaluated using computer tomographic osteoabsorptiometry. Different breeds and species were visually compared. RESULTS: Differences were found in the subchondral bone density distribution of the talus between breeds and between species (Canis familiaris, Canis lupus and Canis aureus). DISCUSSION AND CONCLUSION: Based on the density distribution, there are differences in loading conditions of the tarsocrural joint in different species of Canidae. The joint loading distribution is very similar between dogs of the same breed and within the same species. Although between-breed differences can be explained by conformational differences, the between-species differences remain subject to further research.

Trichinella spp. in wild mesocarnivores in an endemic setting.

Human trichinellosis and Trichinella infection in pigs are both still endemic in the Balkans, including Serbia. Because of the flow between the sylvatic and the domestic cycle of Trichinella spp., monitoring wildlife has been recommended for the risk assessment of Trichinella spp. infection in swine. We have previously shown the presence of Trichinella infection in wild carnivores including the wolf and the golden jackal, and here we report on Trichinella infection in several other mesocarnivore species. From a total of 469 animals collected between 1994 and 2013, Trichinella larvae were detected in 29 (6.2%, 95% CI = 4.0-8.4) animals, including 14 red foxes (4.7%), 7 wild cats (35%), 5 beech martens (4.8%), 2 pine martens (16.7%), and 1 European badger (6.25%). No Trichinella larvae were detected in the examined specimens of European polecats, steppe polecats and European otters. Species identification of the Trichinella larvae performed for 18 positive samples revealed T. spiralis in 77.8% and T. britovi in 22.2% of the isolates. Both species were detected in red foxes and wild cats. The predominance of T. spiralis in wildlife in Serbia indicates the (past or present) spillover of this pathogen from domestic to wild animals.

First evidence of tick-borne protozoan pathogens, Babesia sp. and Hepatozoon canis, in red foxes (vulpes vulpes) in Serbia.

Tick-borne haematozoans cause severe diseases in domestic animals, and some of them have zoonotic potential. The results of previous studies in Europe point to the important role of foxes in natural endemic cycles of several tick-borne pathogens, including protozoa. The aim of the present research was to acquire information on the prevalence and distribution of tick-borne protozoan parasites among foxes in Serbia. Legally hunted foxes from 14 localities throughout Serbia were analysed. Spleen samples were collected from 129 animals and tested for the presence of Babesia spp. and Hepatozoon spp. by PCR. In total, 79/129 (61.2%) of the tested foxes were positive for H. canis, while the presence of two Babesia species was confirmed: B. vulpes (37/129, 28.7%) and B. canis (1/129, 0.8%). Coinfection with B. vulpes and H. canis was present in 26/129 (20.2%) foxes and one animal (1/129, 0.8%) was co-infected by B. canis and H. canis. The results of this study indicate the important role of foxes in the epizootiology of B. vulpes and H. canis in the Republic of Serbia and stress the need for further research to clarify all elements of the enzootic cycle of the detected pathogens, including other reservoirs, vectors, and transmission routes.