Further data on the distribution of Dirofilaria spp. in the Czech Republic in dogs.

Based on previously published data, the Czech Republic is regarded an endemic country of the onchocercid nematodes Dirofilaria immitis (Leidy, 1856) and Dirofilaria repens Railliet et Henry, 1911. Nevertheless, while cases of D. repens are commonly reported from dogs in South Moravia, no recent records of D. immitis are available. Therefore, the present study was performed to clarify the occurrence of both species of Dirofilaria Railliet et Henry, 1910. Blood samples of 551 dogs sampled during 2015 and 2016 were analysed microscopically for presence of microfilariae and blood sera were examined by IDEXX SNAP® 4Dx® test (IDEXX, USA). DNA from blood of microscopically positive dogs was extracted and PCR protocol amplifying fragment of cytochrome c oxidase I (COI) gene was performed; PCR products were then sequenced. All dogs from the Bohemian part of the Czech Republic were negative. The prevalence of D. repens in the Moravian region was 5.7 % (27/476). BLAST analyses of obtained sequences confirmed the presence of D. repens (99-100% identical to KX265049). All sampled animals showed a negative result for D. immitis antigen in IDEXX SNAP® 4Dx® test. Our study confirmed the previously reported occurrence of D. repens in South Moravia and revealed its spreading from the epicentre to the north and west. PCR with subsequent sequencing together with negative results for D. immitis antigen in IDEXX SNAP® 4Dx® test revealed only D. repens infection. A previously published autochthonous infection of dogs with D. immitis in South Moravia was not confirmed.

Eurasian golden jackal as host of canine vector-borne protists.

BACKGROUND: Jackals are medium-sized canids from the wolf-like clade, exhibiting a unique combination of ancestral morphotypes, broad trophic niches, and close phylogenetic relationships with the wolf and dog. Thus, they represent a potential host of several pathogens with diverse transmission routes. Recently, populations of the Eurasian golden jackal Canis aureus have expanded into the Western Palaearctic, including most of Europe. The aim of our study was to examine Eurasian golden jackals from Romania, Czech Republic and Austria for a wide spectrum of vector-borne protists and to evaluate the role of this species as a reservoir of disease for domestic dogs and/or humans. RESULTS: Diagnostic polymerase chain reaction (PCR) DNA amplifications revealed 70% of jackals to be positive for Hepatozoon, 12.5% positive for piroplasms, and one individual positive for Leishmania infantum. Phylogenetic analyses of partial 18S rDNA sequences invariably placed sequenced isolates of Hepatozoon into the H. canis clade. For piroplasms, both the 18S and cox1 sequences obtained confirmed the presence of Babesia canis and "Theileria annae" in 5 and 2 individuals, respectively, providing the first records of these two piroplasmids in Eurasian golden jackals. A single animal from Dolj County (Romania) was PCR-positive for L. infantum, as confirmed also by sequencing of ITS1-5.8S. CONCLUSIONS: Apparently, expanding populations of jackals can play a significant role in spreading and maintaining new Babesia canis foci in Central Europe. The role of jackals in the epidemiology of "Theileria annae" and H. canis is probably similar to that of red foxes and should be taken into account in further research on these parasites. Also the presence of L. infantum deserves attention. Our study confirms that once established, the populations of Eurasian golden jackals constitute natural reservoirs for many canine vector-borne diseases, analogous to the role of the coyotes in North America.

A new case of the enigmatic Candidatus Neoehrlichia sp. (FU98) in a fox from the Czech Republic.

This study reports a new case of Candidatus Neoehrlichia sp. (FU98) infection in a fox from the Czech Republic, and provides confirmatory evidence on the occurrence of this newly identified sequence type. However, further studies are needed to investigate the distribution, host range and possible vector(s) for this bacterium, as well as its impact on animals and humans.

Autochthonous Hepatozoon infection in hunting dogs and foxes from the Czech Republic.

Blood samples from 21 red foxes (Vulpes vulpes) and 8 hunting dogs from the same locality in the Czech Republic were examined for presence of Hepatozoon canis/Hepatozoon sp. The dogs were selected based on their close contact with foxes during fox bolting and because they had not traveled into known endemic areas. Using diagnostic PCR amplifying partial 18S rDNA fragment, Hepatozoon DNA was detected in 20 red foxes (95 %) and 4 dogs (50 %). From 8 positive foxes and 2 positive dogs, we obtained nearly complete 18S rDNA sequences. Phylogenetic analyses of these sequences revealed very low variability. Buffy coat smears from positive dogs were prepared and examined. No Hepatozoon gamonts were found. This study provides the first report of autochthonous infection of H. canis/Hepatozoon in dogs and foxes from the Czech Republic. Our study indirectly demonstrates cross infection between red foxes and dogs and confirms autochthonous infection of Hepatozoon canis in dogs living in a geographic area well outside the range of Rhipicephalus sanguineus sensu lato, which is so far the only known vector of H. canis in Europe.

Dirofilaria repens: emergence of autochthonous human infections in the Czech Republic (case reports).

BACKGROUND: Human dirofilariasis is a zoonotic infection that continues to spread to previously unaffected areas of Europe. In the South Moravian Region of the Czech Republic (CR), imported as well as autochthonous canine infections were recorded in the last decade, and parasite DNA was detected in mosquitoes of Aedes vexans. In the present paper, human Dirofilaria infections are reported from the country for the first time. CASE PRESENTATION: The samples from five patients with suspected tissue helminthiases were investigated. In particular cases, nematodes were isolated from various tissues including skin of lower leg, soft tissues of finger, subcutaneous tissue of hypogastrium, lymph node and peritoneum. The diagnosis was based on light microscopic morphology and/or DNA analysis of the worms. In addition, ELISA examination of patients' sera for anti-filaria IgG antibodies was performed. CONCLUSIONS: In the CR, five cases of human dirofilariasis caused by Dirofilaria repens were recorded during 2010-2014 (species determination for three of them was confirmed besides morphological also by DNA analysis). At least, three of the cases were of autochthonous origin (the patients are Czech citizens residing in South Moravian Region who have never travelled abroad). The findings confirm the natural setting of D. repens in South Moravian Region of the CR. Dirofilariasis should be therefore considered as endemic in this area where it may represent a significant risk factor for public health.

First report of Cercopithifilaria spp. in dogs from Eastern Europe with an overview of their geographic distribution in Europe.

Dogs in Europe may be parasitized by a variety of species of filarioids. While some species have been studied for decades, others have been only recently reported, and their range of distribution is still unknown. The present study was aimed to investigate the occurrence of Cercopithifilaria spp., whose microfilariae reside in the dermis and are transmitted by Rhipicephalus sanguineus s.l. ticks, in a dog population from Romania and to present the current knowledge of their distribution in Europe. Sampling was performed in a rural locality from Danube Delta region of Romania, location chosen due to the presence of the only known vector for these species. Skin samples were collected from 39 dogs. The samples were tested for the presence of Cercopithifilaria spp. by PCR methods. One animal (2.56%) was positive, and the amplified sequence showed a 100% similarity to Cercopithifilaria bainae. This study reports C. bainae for the first time in Eastern Europe, extending the known range of the genus in Europe.