Prevalence of coronaviruses in European bison (Bison bonasus) in Poland.

Coronaviruses have been confirmed to infect a variety of species, but only one case of associated winter dysentery of European bison has been described. The study aimed to analyze the prevalence, and define the impact on the species conservation, the source of coronavirus infection, and the role of the European bison in the transmission of the pathogen in Poland. Molecular and serological screening was performed on 409 European bison from 6 free-ranging and 14 captive herds over the period of 6 years (2017-2023). Presence of coronavirus was confirmed in one nasal swab by pancoronavirus RT-PCR and in 3 nasal swab samples by bovine coronavirus (BCoV) specific real time RT-PCR. The detected virus showed high (> 98%) homology in both RdRp and Spike genes to BCoV strains characterised recently in Polish cattle and strains isolated from wild cervids in Italy. Antibodies specific to BCoV were found in 6.4% of tested samples, all originating from free-ranging animals. Seroprevalence was higher in adult animals over 5 years of age (p = 0.0015) and in females (p = 0.09). Our results suggest that European bison play only a limited role as reservoirs of bovine-like coronaviruses. Although the most probable source of infections in the European bison population in Poland is cattle, other wild ruminants could also be involved. In addition, the zoonotic potential of bovine coronaviruses is quite low.

Antibodies against the Mycobacterium tuberculosis complex and Brucella spp. in captive and free-living European bison (Bison bonasus) in Poland.

BACKGROUND: The European bison (Bison bonasus), a symbol of Polish nature, is a protected species that requires active health monitoring. However, conservation efforts are made difficult by the zoonotic diseases such as brucellosis and tuberculosis. OBJECTIVE: The aim of this study was to screen the Polish European bison population for exposure to the Mycobacterium tuberculosis complex (MTC) and Brucella spp. METHODS: A total of 323 free-living and captive European bison from 13 localities were tested serologically for antibodies against the M. bovis P22 multi-protein complex (in-house ELISA) and against Brucella spp. (commercial ELISA). RESULTS: Antibodies against the MTC (P22) were detected in 7% (22/323) of the tested European bison. Anti-MTC antibody positivity was not significantly different by sex, age, and captive/free range status. Anti-MTC antibodies were found in six of 13 populations sampled, always in populations with larger sample sizes including the four free-living ones. Antibodies against Brucella spp. were detected in 36% (116/323) of the tested bison. While Brucella spp. antibody prevalence was not different by sex, it was significantly different by age (lower in adults) and captive/free-living status. Brucella spp. seroprevalence decreased with sample size and seropositive bison were found in 12 of 13 sampling populations. CONCLUSIONS: Our findings identify potential emerging threats to the European bison population and confirm the first serological response to P22 in European bison. As Poland is currently officially free of brucellosis and bovine tuberculosis, our results require careful interpretation. Further studies are needed to establish the presence of cross-reactions with atypical mycobacteria in the case of MTC and other bacteria (e.g. Yersinia enterocolitica O:9) in the case of Brucella spp.

Morphometric and Molecular Analysis of Five-Spine Epidinium Morphotypes Taken from the Rumen of European Bison, Bison bonasus.

An important feature of ruminal ciliates is their phenotypic plasticity, which makes their identification difficult. The common manifestation of the phenotypic plasticity in rumen ciliates is a change in their cell size and caudal spination. We analyzed various morphotypes of Epidinium with five caudal processes (spines) taken from the rumen of European bison (Bison bonasus). In the study, the cluster analysis and K-means analysis of morphometric data could not distinguish very similar morphotypes of Epidinium with five caudal processes. However, the morphotype of E. parvicaudatum prevailed (70%). The DNA of four individual E. parvicaudatum was isolated successfully from formaldehyde-preserved samples. The partial 18S rDNA gene sequences (about 350-400 bp) were identical to Epidinium sequences in GenBank (E. caudatum, a one-spine morphotype, and E. cattanei, a five-spine morphotype). It can be assumed that these short sequences cannot distinguish the differences between the Epidinium morphospecies. Complete gene sequences from various hosts and various molecular markers are necessary to reveal the validity of the Epidinium five-spine species. In conclusion, classical morphology should be supplemented with molecular data when more morphotypes of the rumen ciliate species are present in samples.

A serological survey of pathogens associated with the respiratory and digestive system in the Polish European bison (Bison bonasus) population in 2017-2022.

BACKGROUND: The European bison (Bison bonasus) is a near threatened species and requires health monitoring. The aim of the present study was to determine the prevalence of antibodies to pathogens known to cause respiratory and digestive illness in ruminants. RESULTS: In the studied 328 European bison, the highest seroprevalence was observed for Bovine herpesvirus-1 (BoHV-1) (50.27%), Bovine Coronavirus (BCoV) (26.36%), and Bluetongue Virus (BTV) (12.83%). For Mycoplasma bovis strains and Bovine Viral Diarrhea Virus (BVDV), positive results were rare. Interestingly, a higher prevalence of BTV antibodies was noted in the northeastern populations and older animals. CONCLUSIONS: Our findings indicate that the Polish European bison population appears to have considerable contact with BoHV-1; however, this does not appear to be of great significance, as clinical symptoms and post-mortem lesions are rarely noted in Polish European bison population. The high seroprevalence of BTV in the north-east of Poland is an ongoing trend, also noted in previous studies. It is possible that European bison may perpetuate the virus in this region. This is the first report of antibodies for BCoV in European bison.

Effect of Translocation on Host Diet and Parasite Egg Burden: A Study of the European Bison (Bison bonasus).

For the purpose of nature management and species conservation, European bison (Bison bonasus) are being increasingly reintroduced into nature reserves across Europe. The aim of this study was to investigate European bison's adaptability to new areas through the study of their parasite-EPG (eggs per gram feces) and dietary diversity during twelve months after translocation. We compared the parasite-EPG from introduced European bison in Lille Vildmose, Denmark, with the parasite-EPG from populations from Bornholm, Denmark, and Bialowieza Forest, Poland. From March 2021 to February 2022, fecal samples were collected from three populations. Samples from Lille Vildmose were examined through flotation, sedimentation, the Baermann technique, and nanopore sequencing. Fecal samples from Bornholm and Bialowieza were examined through flotation and sedimentation. Nanopore sequencing of DNA from 63 European bison's fecal samples collected during March-September in Lille Vildmose identified 8 species of nematodes within the digestive tract of the European bison, with Haemonchus contortus being the most frequently observed. In Lille Vildmose, a significantly higher excretion of nematode-EPG was observed during the summer period than in the spring, autumn, and winter. In addition, monthly differences in the excretion of nematode eggs were found, with this being significantly higher in June than in the months during autumn and winter (October-February). Significant differences in the nematode-EPG were only found between the excretion of nematode eggs in Bialowieza Forest when compared to that of Lille Vildmose, with significantly higher excretion in Lille Vildmose (October-November). The results indicate that the development rates for nematodes may be affected by changes in temperature, with increasing temperatures speeding up their development time. Independent of this study design, wildlife vets together with the gamekeepers managing the herd found it necessary to treat the herd with antiparasitics for practical and animal welfare reasons in relation to translocation. Furthermore, 79 plant taxa were identified in the diet of the European bison. The broadest diet was observed in March suggesting that the European bison quickly adapted to their new habitat. The results suggest a seasonal shift in their diet, with this being most apparent from March to April.

Ten Years of Animal Tuberculosis Monitoring in Free-Living European Bison (Bison bonasus) in Poland.

In the period 1996-2012, two outbreaks of animal tuberculosis were noted in the population of free-living European bison (Bison bonasus caucasicus) in the Bieszczady Mountains, Southern Poland. As the European bison is an endangered species and particularly susceptible to tuberculosis, not to mention a national icon, the decision was made to test all deceased bison for TB in Poland. The screened bison were obtained by elimination due to poor health or natural death. A total of 159 European bison have been examined over the last 10 years. The individuals came from four regions of Poland (Bialowieza Forest, Bieszczady Mountains, Borecka Forest, Knyszynska Forest), not only from the area where tuberculosis is still endemic. Mycobacterium bovis and Mycobacterium avium spp. hominisuis were identified in two different herds. The isolation of M. bovis from European bison was the first case described in Poland. So far, the only causative agent of tuberculosis identified in European bison in Poland, both in the wild and in captive herds, was Mycobacterium caprae. The isolated M. bovis spoligotype has not previously been registered in international spoligotype databases so far. The obtained results highlight the need to monitor TB in European bison in Poland.

Is Serological Monitoring a Fit-for-purpose Tool to Assess the Epidemiological Situation of Tuberculosis in the Sylvatic Species of European Bison (Bison Bonasus) in Poland?

Introduction: Bovine tuberculosis is one of the most dangerous zoonotic diseases. Despite the near-complete elimination of the disease from cattle breeding in Poland achieved in 2009, its re-emergence is now observed. Globally, the number of human cases is underestimated and the importance of free-living animals as reservoirs of tuberculosis is growing. As a species highly susceptible to Mycobacterium tuberculosis complex infection, the European bison (Bison bonasus) has a role in the transmission of the disease in Poland. The purpose of the investigation was to assess the epidemiological situation of tuberculosis in Polish European bison serologically. Material and Methods: A total of 460 serum samples were collected from 436 European bison from 15 out of 26 national populations between 2013 and 2020. An M. bovis ELISA was used, and its sensitivity and specificity were assessed with an eyelid tuberculin skin test (TST) and interferon gamma release assay (IGRA). Results: Mycobacterium bovis antibodies were detected in nine serum samples. The presence of antibodies was found in two animals from the Bialowieza Forest (1.2% of the population), and one each from the Borecka Forest (2.4%) and the Warsaw Zoo (14.3%). One European bison among the 14 sampled (7.1%) from Smardzewice was positive on five occasions. Other samples from Smardzewice and the Bieszczady Mountains, where tuberculosis had previously been reported, were negative. Conclusion: ELISA testing is an effective, easy and cost-efficient tool for monitoring of tuberculosis-naïve populations. Serological testing in tuberculosis control programmes can significantly improve the detection of infected herds. Antibody ELISAs may supplement TST and IGRA, but cannot replace them.

Three new genes associated with longevity in the European Bison.

Longevity-related genes have been found in humans, mice, dogs and in several other animal species. The goal of this study was to perform genetic analysis of long-lived European bisons with the aim to find genes that are associated with longevity using GWAS and further sequencing of a wider sample panel. European bison has a unique history of near extinction and the recovery of the species from just a few founder individuals. Together with the short medium lifespan, the expected genetic homogeneity makes bison a suitable model for studying longevity. Particular single nucleotide polymorphisms within three genes, BCKDHB, FER1L6 and SERPINI2, were found significantly overrepresented in long-lived European bisons. In SERPINI2, the longevity-associated single nucleotide polymorphism localizes to an exon. In the protein encoded by the SERPINI2 gene, amino acid leucine present in the reference European bisons is replaced by tryptophan in the long-lived animals. This study is the first to determine longevity-associated variants in genes in European bison. Association of the FER1L6 gene with longevity shows a possible sex dependency.

Prevalence and Genetic Diversity of Trueperella pyogenes Isolated from Infections in European Bison (Bison bonasus).

Trueperella pyogenes is a Gram-positive bacterium causing purulent infections in many animal species, including the European bison. However, the data about the virulence and genetic relationships of T. pyogenes strains isolated from these wild ruminants are strongly limited. The aim of this study was to investigate the prevalence of T. pyogenes infections in the European bison, and to evaluate the genetic diversity of isolates from these animals. In the time span of 10 years, 328 European bison from 16 different locations were examined. The standard bacteriological methods were used for T. pyogenes isolation and identification from clinical specimens obtained from urogenital tract infections and abscesses of different locations. The presence of genes encoding known virulence factors was investigated by PCR, and the genetic diversity of T. pyogenes strains was examined with the RAPD-PCR method. The prevalence of T. pyogenes infections was 14.6%, and the pathogen was isolated from both female (47.9% of isolates) and male (52.1% of isolates) European bison. It should be highlighted that a considerable number of strains were isolated from the prepuce and penis infections. Therefore, the role of T. pyogenes in the pathogenesis of balanoposthitis should be seriously perceived. A total of 39 T. pyogenes strains were subjected to genetic characterization. All studied strains carried the plo gene, while the nanH (25.6%), nanP (23.1%), cbpA (7.7%), fimA (97.4%), fimC (69.2%), fimE (92.3%) and fimG (15.4%) genes were present with a variable frequency among the tested strains. The virulence genotype plo/fimA/fimC/fimE was dominant. RAPD-PCR typing showed a high level of genetic diversity among European bison T. pyogenes strains, and a total of 31 different RAPD profiles were distinguished. In a few cases, the same RAPD profile was found in strains obtained from animals living in the same area. This study provided the first data about the prevalence and genetic relationships of T. pyogenes in the Polish population of European bison. However, further epidemiological investigations are needed to understand the routes of transmission and dissemination of the pathogen in these wild animals.

Pregnancy in European bison (Bison bonaus) with generalized tuberculosis - no evidence of vertical transmission.

INTRODUCTION AND OBJECTIVE: One of the main health threats to the endangered European bison (Bison bonsasus) is bovine tuberculosis, the pathogenesis of which in this species is not fully known. The aim of the study was to confirm a possible case of vertical transmission from a pregnant European bison with generalized tuberculosis to its 12-week-old foetus. MATERIAL AND METHODS: During the autopsy it was found that the bison had become pregnant, despite an advanced stage of tuberculosis. Material collected from the organs and foetus was placed on Lowenstein and Stonebrink media and incubated at 37 °C for 12 weeks. RESULTS: Mycobacteria were isolated from the lungs and lymph nodes; however, the tissue of the foetus and fragments of the reproductive system were negative. CONCLUSIONS: Vertical transmission was excluded, although it cannot be ruled out that infection could occur as pregnancy progresses.

Establishment of a Wisent (Bison bonasus) Germplasm Bank.

The wisent, or European bison (Bison bonasus), belongs to the same family (Bovidae) as the American bison and domestic cattle. The wisent is the largest mammal in Europe, and is called the "Forest Emperor". The wisent is listed as "Vulnerable" on the IUCN Red List, and is protected by international law. Achievements in reproductive biotechnology have opened new possibilities for the cryoconservation of the wisent germplasm. Therefore, this research aimed to improve a strategy for the protection and preservation of the European bison through the creation of a wisent germplasm bank, based on the following procedures: isolation and in vitro maturation (IVM) of oocytes, in vitro fertilization (IVF) of matured oocytes, in vitro embryo culture (IVC), and embryo cryopreservation. Wisent ovaries were isolated from females outside the reproductive season, and eliminated from breeding for reasons other than infertility. Cumulus-oocyte complexes (COCs) were isolated from follicles greater than 2 mm in diameter and matured for 24 h and 30 h. After IVM, COCs were fertilized in vitro with wisent sperm. The obtained wisent zygotes, based on oocytes matured for 24 h and 30 h, were cultured for 216 h. Embryos at the morula and early blastocyst stages were vitrified and then warmed and transferred to interspecies recipients (Bos taurus). USG and biochemical tests were used to monitor pregnancies. This study obtained embryos in the morula and early blastocyst stages only after oocytes were fertilized and matured for 30 h. On average, per oocyte donor, 12.33 ± 0.5 COCs were isolated, and only 9.33 ± 0.61 COCs were qualified for in vitro maturation (75.68%), while 9.16 ± 0.48 COCs were matured (84.32%). On average, per donor, 5.5 ± 0.34 embryos were cleaved (59.96%) after 48 h post-fertilization (hpf), and 3.33 ± 0.21 achieved the eight-cell stage (36.52%) after 96 hpf, while 1 ± 0.21 morula and early blastocyst stages (10.71%) were achieved after 216 hpf. A total of six embryos (one morula and five early blastocysts) were obtained and vitrified; after warming, five of them were interspecies transferred to cattle (Bos taurus). On day 41 after fertilization, 3 out of 5 pregnancies were detected based on USG, P4, and PAG tests. However, no pregnancy was observed on day 86 after fertilization, indicating embryo resorption. This study shows that obtaining wisent embryos in vitro, and subsequent cryopreservation to create a wisent embryo bank, can be applied and implemented for the wisent protection program.

Case Report: Proximal Phalangeal Fracture Management in a European Bison (Bison bonasus).

Fracture of the digits is a well-known orthopedic condition in adult cattle, and mainly the distal phalanx (P3) is involved. To our knowledge, the treatment of fractures of the middle (P2) and proximal (P1) phalanges with orthopedic claw blocks has not yet been described in cattle and other ruminants. This report describes the first case of the successful management of a P1 fracture in an adult European bison. A 5-year-old female European bison (Bison bonasus) presented with severe weight bearing lameness of the left hind limb and a marked soft tissue swelling accentuated over the plantar and lateral aspects of the proximal and middle phalanges, associated with multifocal light bluish discoloration of the skin (hematoma) and increased local temperature. The cow was examined and managed because of a severely comminuted fracture of the lateral P1 of the left hind limb. Application of an orthopedic block on the healthy medial partner claw allowed to reduce the load of the affected digit. Combined with the administration of non-steroidal anti-inflammatory drugs, it supported immobilization and recovery. Radiographic re-evaluations at weeks 6, 9, and 11 after the injury revealed progressive callus formation and fracture consolidation. From week 9 onwards, until the end of treatment, no lameness was observed. The healing process was good, and both clinical and radiographical improvement were evident after immobilizing the affected digit by applying an orthopedic claw block on the healthy partner claw and administering non-steroidal anti-inflammatory drugs.

Intra-Palpebral Tuberculin Skin Test and Interferon Gamma Release Assay in Diagnosing Tuberculosis Due to Mycobacterium caprae in European Bison (Bison bonasus).

Despite the threat posed by tuberculosis (TB) to the protected European bison (Bison bonasus), no validated TB tests exist for this species. This pilot study evaluates two tests based on detecting cellular immunity for this purpose: interferon gamma release assay (IGRA) and tuberculin skin test (TST). Ten animals were subjected to ante-mortem and post-mortem examinations. IGRA was performed using a commercial test, and the comparative TST was performed in the eyelids. The lesions were assessed post-mortem and material was collected for mycobacterial culture. The isolated strains were subjected to genotyping. At post-mortem examination, five out of ten individuals demonstrated both tuberculous lesions and positive culture results (Mycobacterium caprae). Compared to the palpebral TST, the findings of the IGRA are easier to interpret when diagnosing tuberculosis in European bison.

Veterinary monitoring of gastrointestinal parasites in European bison, Bison bonasus designated for translocation: Comparison of two coprological methods.

It is important to monitor the level of parasitic loads in herds of European bison and to identify threats early enough to prevent their spread to other populations or species. The aim of the present study was to compare the detection sensitivity of two fecal flotation techniques, viz. the modified Willis method (WM) with centrifugation and modified McMaster flotation technique (MM), in the diagnostics of gastrointestinal parasites of European bison before the translocation of animals. Out of 166 feces samples, Eimeria spp. oocysts (84.3% in WM and 71.1% in MM) and Trichostrongylidae eggs (82.5% in WM and 53.6% in MM) predominated. These were accompanied by eggs from Capillaria spp. (prevalence: 13.9% in WM and 3.61% in MM), Nematodirus spp. (prevalence: 18.1% in WM and 4.8% in MM) and Trichuris spp. (prevalence: 12.7% in WM and MM) were identified. The lowest prevalence was noted for cestode eggs of Moniezia spp. (5.4% in WM and 3.0% in MM). The Willis method yielded a higher prevalence of eggs and oocysts than the modified McMaster method, and hence has a higher probability of detecting parasitic structures than the modified McMaster method, especially in cases of very low levels of invasion. As the two methods yield consistent results, it is recommended to use the Willis method for diagnosis of internal parasite infection in European bison. This test offers more sensitive method than McMaster technique of detecting the presence of low levels of a variety of parasite eggs and oocysts in feces, while also being inexpensive and adaptable to field work.

Three-dimensional characteristic of fungiform papillae and its taste buds in European bison (Bison bonasus), cattle (Bos taurus), and Bison bonasus hybrid.

BACKGROUND: Our recent macro- and scanning electron microscopic study of tongue conducted on domesticated cattle, wild living European bison, and Bison bonasus hybrid revealed various spatial arrangement and number of gustatory and mechanical papillae between parental species and their hybrid. Furthermore, scanning electron microscopy analysis of gustatory papillae indicated the variable distribution of fungiform papillae (Fu) over the surface of the tongue, which could be significant in differentiated taste perception during feeding in studied wild living and domesticated husbandry ruminants. To specify the detailed microstructure of Fu papillae with connective tissue cores (CTC) and intraepithelial taste buds system, the first time the three-dimensional computer-aided analysis of serial histoslides resulted in the rendering of 3D reconstructions of Fu papillae. RESULTS: The comparative analysis of 3D models Fu papillae conducted in six areas of lingual mucosa of each tongue revealed information about, microstructural diversity of Fu papillae in studied ruminants. The estimation of number and density of Fu papillae on tongues, rate of protrusion of papillae over mucosa, and a number of taste buds per papilla allowed to state the ventral surface of the lingual apex and posterolateral surfaces of the lingual torus as regions important in taste perception, as in the preselection of taken food, as well in the analysis of food during rumination, respectively. On the 3D models were observed three structural types of CTC of different distribution on the tongue in studied species. The quantitative data of the number of taste buds on Fu papillae have regional functional differences in the taste system important in feeding and veterinary practice. Moreover, our analysis determined specific features in examined hybrid and showed similarities of some studied features with cattle, i.e., maternal species. CONCLUSIONS: The 3D reconstruction method used for the first time in the field of study of the lingual papillae and taste buds system can be considered as an innovative and effective tool in assessing of the microstructures of Fu papillae, and it could be suitable for further studies of taste system structures in normal and pathological condition.

Benzimidazole resistant Haemonchus contortus in a wildlife park.

INTRODUCTION: Anthelmintic resistance (AR) of ruminant gastrointestinal nematodes (GINs) constitutes a major problem worldwide. Among GINs, the abomasal blood-feeding parasite Haemonchus contortus is particularly pathogenic and may show resistance against all major anthelmintic substance classes. In the present study, the death of a 1,5 year-old European bison (Bison bonasus) from a German wildlife park due to haemonchosis despite frequent anthelmintic treatment of the herd with fenbendazole as well as doramectin prompted an investigation regarding AR. Pooled faecal samples were collected from four different bison groups as well as from mouflons (Ovis orientalis), elk (Alces alces), reindeer (Rangifer tarandus), sika deer (Cervus nippon), Persian fallow deer (Dama mesopotamica) and red deer (Cervus elaphus) housed by the wildlife park. After coproscopical examination, faecal larval cultures were established. Haemonchus contortus-positive larval cultures were further examined for genetic polymorphisms associated with benzimidazole resistance at codons 167 and 200 of the ß-tubulin isotype 1 gene by real-time pyrosequencing. Infections with H. contortus were detected in all four bison groups, as well as in mouflons. In five samples, representing two bison groups and the mouflons, the frequency of the resistance-associated single-nucleotide polymorphism (SNP) at codon 200 was 100%. In contrast, resistance-associated SNPs were not detected at codon 167. In addition, faecal egg counts from two bison before and 14 days after parenteral doramectin treatment indicated possible macrocyclic lactone resistance. Detection of anthelmintic resistant nematodes in these animals was especially concerning in the light of planned reintroduction into the wild. As helminth control in zoological gardens and wildlife parks relies mostly on anthelmintic treatment due to restricted possibilities regarding management practices such as rotational grazing, care should be taken to avoid underdosing or unnecessary frequent treatments facilitating the development of AR.

INTRODUCTION: La résistance aux anthelminthiques (RA) des nématodes gastro-intestinaux (NGI) des ruminants constitue un problème majeur dans le monde. Parmi les NGI, le parasite hématophage de la caillette Haemonchus contortus est particulièrement pathogène et peut montrer une résistance contre toutes les principales classes de substances anthelminthiques. Dans la présente étude, la mort d'un bison d'Europe de 1,5 ans (Bison bonasus) d'un parc animalier allemand en raison d'une hémonchose malgré un traitement anthelminthique fréquent du troupeau avec du fenbendazole ainsi que de la doramectine a incité une enquête sur la RA. Des échantillons de matières fécales regroupés ont été collectés auprès de quatre groupes de bisons différents ainsi que de mouflons (Ovis orientalis), de wapitis (Alces alces), de rennes (Rangifer tarandus), de cerfs sika (Cervus nippon), de daims de Perse (Dama mesopotamica) et de cerfs élaphes (Cervus elaphus) hébergés par le parc animalier. Après examen coproscopique, des cultures larvaires fécales ont été réalisées. Des cultures larvaires positives pour Haemonchus contortus ont été examinées plus avant pour les polymorphismes génétiques associés à la résistance au benzimidazole aux codons 167 et 200 du gène de l'isotype 1 de la ß-tubuline par pyroséquençage en temps réel. Des infections à H. contortus ont été détectées dans les quatre groupes de bisons, ainsi que chez les mouflons. Dans cinq échantillons, représentant deux groupes de bisons et les mouflons, la fréquence du polymorphisme mononucléotidique (SNP) associé à la résistance au codon 200 était de 100 %. En revanche, les SNP associés à la résistance n'ont pas été détectés au codon 167. De plus, le nombre d'œufs par gramme de fèces de deux bisons avant et 14 jours après le traitement parentéral à la doramectine a indiqué une possible résistance aux lactones macrocycliques. La détection de nématodes résistants aux anthelminthiques chez ces animaux était particulièrement préoccupante à la lumière de la réintroduction prévue dans la nature. Étant donné que le contrôle des helminthes dans les jardins zoologiques et les parcs animaliers repose principalement sur un traitement anthelminthique en raison des possibilités limitées concernant les pratiques de gestion telles que le pâturage en rotation, des précautions doivent être prises pour éviter un sous-dosage ou des traitements fréquents inutiles facilitant le développement de la RA.

Demodex bialoviensis sp. nov. (Acariformes, Demodecidae) a new, specific parasite of the European bison Bison bonasus (Artiodactyla, Bovidae).

Sixteen species of parasitic mites of the family Demodecidae have been described in ungulates (Ungulatomorpha), particularly among domestic animals and livestock. Specific synhospital species have been found in seven host species. Demodex bisonianus, occurring in the Meibomian glands of the eyelids, was described in the European bison Bison bonasus. Together with Trypanosoma wrublewskii and Bisonicola sedecimdecembrii, it is one of the three known specific parasite species of European bison. The European bison is not only a rare and endangered mammal, but its biology has been shaped by its recent restitution from extinction from a small pool of individuals kept in breeding facilities. This could have been the reason for the extinction of the majority of specific parasites. The present study reports the finding of a new specific parasite species, Demodex bialoviensis sp. nov., associated with the nasal skin region, in European bison from the Bialowieza Forest (Poland). It is the second species of Demodex reported in this host species, however occurring in a different location.

Acute-phase protein concentrations in serum of clinically healthy and diseased European bison (Bison bonasus) - preliminary study.

BACKGROUND: This is the first report describing levels of APPs in European bison. Serum concentration of acute phase proteins (APPs) may be helpful to assess general health status in wildlife and potentially useful in selecting animals for elimination. Since there is a lack of literature data regarding concentration of APPs in European bisons, establishment of the reference values is also needed. METHODS: A total of 87 European bison from Polish populations were divided into two groups: (1) healthy: immobilized for transportation, placing a telemetry collar and routine diagnostic purposes; and (2) selectively culled due to the poor health condition. The serum concentration of haptoglobin, serum amyloid A and α1-acid-glycoprotein were determined using commercial quantitative ELISA assays. Since none of the variables met the normality assumptions, non-parametric Mann-Whitney U test was used for all comparisons. Statistical significance was set at p < 0.05. Statistical analyses were performed using Statistica 13.3 (Tibco, USA). RESULTS: The concentration of haptoglobin and serum amyloid A was significantly higher in animals culled (euthanised) due to the poor condition in respect to the clinically healthy European bison. The levels of α1-acid-glycoprotein did not show statistical difference between healthy and sick animals. CONCLUSIONS: Correlation between APPs concertation and health status was proven, therefore the determination of selected APPs may be considered in future as auxiliary predictive tool in assessing European bison health condition.

Microbiological and molecular monitoring for bovine tuberculosis in the Polish population of European bison (Bison bonasus).

INTRODUCTION AND OBJECTIVE: In recent years, bovine tuberculosis (BTB) has become one of the major health hazards facing the European bison (EB, Bison bonasus), a vulnerable species that requires active protection, including regular and effective health monitoring. Monitoring of zoonotic disease in wildlife is also an important part of public health protection. The aim of the study was to determine whether BTB still influences the EB population in Poland. MATERIAL AND METHODS: During 2017-2019, mandibular, retropharyngeal and mediastinal lymph nodes were collected from 90 EB during post-mortem examination, and then cultivated on Lowenstein-Jensen and Stonebrink media. Isolated strains were subjected to molecular analysis to determine the species, spoligotype and MIRU-VNTR pattern. RESULTS: Lesions were found in lymph nodes originating from eight EB (8.89%). Positive microbiological cultures for mycobacteria were obtained in samples from six (6.67%) EB. The isolated strains were identified as Mycobacterium caprae (material from four EB) and atypical mycobacteria (material from two EB). For M. caprae strains spoligotype M. bovis 4_CA 1600 was identified and the MIRU-VNTR pattern was identified as 345751355413232. CONCLUSIONS: It is recommended that this potentially dangerous disease should be monitored in EB via a comprehensive strategy based on a combination of microbiological and molecular methods. Such monitoring will protect the health of both animals and humans.

Pathogens with potential impact on reproduction in captive and free-ranging European bison (Bison bonasus) in Poland - a serological survey.

BACKGROUND: The European bison is an endangered species, and as such it is extremely important to monitor herds for pathogens which can lead to reproductive failure. The aim of the present study was to determine the current prevalence of antibodies to pathogens known to potentially influence reproduction in European bison. Serum samples from 183 bison, originating from different parts of Poland, were tested using commercial ELISA tests for antibodies to Chlamydia spp., Coxiella burnetti, Leptospira interrogans, Neospora caninum and Toxoplasma gondii; the findings were compared between captive and main free-ranging herds, and with regard to the influence of demographic factors such as age and sex. The prevalence of seropositivity was also checked with regard to location and the animal species sharing it. RESULTS: Chlamydia spp. antibodies were present in 48 out of 130 (36.9%) tested samples. Coxiella burnetii was found in one sample out of 178 (0.58%). N. caninum in 36 out of 172 (20.9%) and T. gondii in 23 out of 172 (13.4%). No sample was positive for leptospirosis. Neither sex nor age appeared to have a significant effect on the occurrence of antibodies to the identified species. The prevalence of Chlamydia spp. in the samples varied significantly according to location; however, similar frequency ranges were observed between free ranging and captive herds. In contrast, antibodies to N. caninum were more common in free-ranging herds than captive herds, with the highest frequency observed in the Bieszczady Mountains. CONCLUSIONS: Chlamydia spp., N. caninum and T. gondii might have a similar impact on the reproductive potential of European bison as they have on cattle. The high occurrence of antibodies to N. caninum in bison from the Bieszczady Mountains may be associated with the relatively high density of the wolf population in the area.