High frequency of benzimidazole resistance alleles in trichostrongyloids from Austrian sheep flocks in an alpine transhumance management system.

BACKGROUND: Infections of small ruminants with trichostrongyloid nematodes often result in reduced productivity and may be detrimental to the host. Anthelmintic resistance (AR) against most anthelmintic drug classes is now widespread amongst the trichostrongyloids. Baseline establishment, followed by regular monitoring of the level of AR, is necessary for farmers and veterinarians to make informed decisions about parasite management. The detection of single nucleotide polymorphisms (SNPs) is a sensitive method to detect AR against benzimidazoles (BZs), one of the most widely used anthelmintic classes. Alpine transhumance constitutes a special type of pasturing of sheep from many different farms, the aim of this study was to investigate the prevalence of benzimidazole resistance alleles in this particular management system. RESULTS: Sixteen sheep flocks in Styria and Salzburg in Austria were examined by pyrosequencing for SNPs at codons 167, 198 and 200 of the isotype-1 ß-tubulin gene. The frequency of the resistance-associated exchange F200Y was 87-100% for H. contortus, 77-100% for T. colubriformis and <  5-66% for T. circumcincta. Additionally, the F167Y polymorphism was detected in T. colubriformis from two farms at a frequency of 19 and 23% respectively. CONCLUSIONS: The high resistance allele frequency in H. contortus and T. colubriformis in the examined sheep population urgently calls for the development of new treatment strategies to sustainably control trichostrongyloid infections for this kind of pasturing, since the frequent mixing of flocks during the alpine summer grazing must be considered an important risk factor for the spread of resistant nematodes to a large number of farms.

Adult Hyalomma marginatum tick positive for Rickettsia aeschlimannii in Austria, October 2018.

We report on a non-indigenous adult Hyalomma marginatum tick in Austria carrying the human pathogenic Rickettsia aeschlimannii; presumably introduced as a nymph via migratory birds and completed the moulting within the same year. It was negative for Crimean-Congo haemorrhagic fever virus, but the finding of R. aeschlimannii represents a potential threat for humans due to its zoonotic character. Awareness of invasive tick species and carried pathogens should be improved in central and northern Europe.

A molecular survey of vector-borne pathogens in red foxes (Vulpes vulpes) from Bosnia and Herzegovina.

BACKGROUND: Red foxes (Vulpes vulpes) have recently been recognized as potential reservoirs of several vector-borne pathogens and a source of infection for domestic dogs and humans, mostly due to their close vicinity to urban areas and frequent exposure to different arthropod vectors. The aim of this study was to investigate the presence and distribution of Babesia spp., Hepatozoon canis, Anaplasma spp., Bartonella spp., 'Candidatus Neoehrlichia mikurensis', Ehrlichia canis, Rickettsia spp. and blood filaroid nematodes in free-ranging red foxes from Bosnia and Herzegovina. METHODS: Spleen samples from a total of 119 red foxes, shot during the hunting season between October 2013 and April 2014 throughout Bosnia and Herzegovina, were examined for the presence of blood vector-borne pathogens by conventional PCRs and sequencing. RESULTS: In the present study, three species of apicomplexan parasites were molecularly identified in 73 red foxes from the entire sample area, with an overall prevalence of 60.8%. The DNA of B. canis, B. cf. microti and H. canis was found in 1 (0.8%), 38 (31.9%) and 46 (38.6%) spleen samples, respectively. In 11 samples (9.2%) co-infections with B. cf. microti and H. canis were detected and one fox harboured all three parasites (0.8%). There were no statistically significant differences between geographical region, sex or age of the host in the infection prevalence of B. cf. microti, although females (52.9%; 18/34) were significantly more infected with H. canis than males (32.9%; 28/85). The presence of vector-borne bacteria and filaroid nematodes was not detected in our study. CONCLUSION: This is the first report of B. canis, B. cf. microti and H. canis parasites in foxes from Bosnia and Herzegovina and the data presented here provide a first insight into the distribution of these pathogens among the red fox population. Moreover, the relatively high prevalence of B. cf. microti and H. canis reinforces the assumption that this wild canid species might be a possible reservoir and source of infection for domestic dogs.

Acute granulocytic anaplasmosis in a captive timber wolf (Canis lupus occidentalis).

A 2-yr-old male captive timber wolf, kept in an outdoor enclosure in Austria, presented with anorexia, depression, and fever in June. Tick infestation was reported despite monthly acaricidal treatment. The microscopic examination of a blood smear revealed elementary bodies and morulae suspicious for Anaplasma phagocytophilum. Polymerase chain reaction confirmed the infection by the G-variant of the pathogen, also found in horses and humans but not in wild ruminants. The wolf seroconverted within 2 wk, and antibodies persisted thereafter. Therapy was started with doxycycline for 10 days, and the wolf recovered within 24 hr. Clinical symptoms and temporary changes in blood parameters (thrombocytopenia, lymphopenia, mild anemia) in this wolf show high similarity to granulocytic anaplasmosis seen in dogs. This is the first report on granulocytic anaplasmosis in a wolf, indicating that A. phagocytophilum might cause clinical disease in the wolf.

[Detection of Dirofilaria in Austrian dogs]. / Nachweis von Dirofilarien in österreichischen Hunden.

A recent increase in the occurrence of Dirofilaria repens and Dirofilaria immitis has been reported from the eastern neighbouring countries of Austria (Slovakia, Hungary). In order to obtain data from Austria--especially from the areas bordering these countries--blood samples of dogs from Gänserndorf (northeastern Austria, bordering Slovakia) and Neusiedl (eastern Austria, bordering Hungary) were examined for the presence of Dirofilaria spp.-DNA by quantitative real-time PCR (qPCR). For D. repens one (out of eight) and six (out of ninety) positive dogs were found, respectively. D. immitis-DNA was not detected. Most of the positive dogs had previously been abroad in known Dirofilaria endemic areas. Two dogs, however, had never been abroad and presumably became infected in Austria. To investigate an autochthonous focus, 1366 mosquitoes from the eastern part of Austria were also examined by qPCR. Although positive mosquitoes could not be found, an emerging endemic occurrence of D. repens in Austria due to positive dogs seems possible.

Parasites on paper--The use of FTA Elute((R)) for the detection of Dirofilaria repens microfilariae in canine blood.

Storage of blood samples for subsequent DNA extraction without loss of integrity can be difficult under field conditions. Filter-based technologies are known to deliver good results for viral, bacterial and protozoan material but have not been tested for blood-dwelling stages of nematodes. In this study Whatman FTA Elute (Whatman plc, Middlesex, UK) filter technology was tested for its ability to stabilise DNA from blood samples of dogs naturally infected with Dirofilaria repens for storage prior to PCR. The concentration of microfilariae per 100microl of blood was evaluated and the blood was diluted to determine the lowest detectable number of parasites by real-time PCR (qPCR). In parallel, negative dog blood was prepared in the same way. A parasitaemia of 6+/-0.43 larvae per 100microl of blood containing EDTA could be detected using the FTA Elute filter cards, although quantification of the larvae by using the qPCR was not possible. Inasmuch as the average microfilaremia in a pool of positive dogs was 311+/-21.72 larvae per 100microl, these cards provide an effective tool for parasitological field studies because blood samples are stable for extended periods of time at room temperature without loss of DNA integrity.