Adnan Hodžić

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

BackgroundRed 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.MethodsSpleen 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.ResultsIn 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.ConclusionThis 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.

Candidatus Neoehrlichia sp. in an Austrian fox is distinct from Candidatus Neoehrlichia mikurensis, but closer related to Candidatus Neoehrlichia lotoris

BackgroundCandidatus Neoehrlichia came under the focus of recent research in terms of human and pet relevance. Candidatus Neoehrlichia mikurensis seems to be relatively abundant in animals and humans from Central European countries, whereas Candidatus Neoehrlichia lotoris was found solely in raccoons from the USA.FindingsSpleen samples from a total of 164 red foxes, originating from two western provinces in Austria (Tyrol and Vorarlberg), were collected and examined for the presence of tick-borne bacteria of the family Anaplasmataceae by PCR and sequencing. In a fox sample originating from Vorarlberg Candidatus Neoehrlichia sp. was found, which is genetically (16S rRNA, groEL) closely related to Candidatus Neoehrlichia lotoris but clearly distinct from Candidatus Neoehrlichia mikurensis.ConclusionsThe present study revealed, for the first time, the occurrence of Candidatus Neoehrlichia sp. in a red fox worldwide. A continuing screening of wild carnivores, especially foxes, and ticks for this potential pathogen is required to evaluate the actual occurrence and distribution of these bacteria. Further research is needed to elucidate the relationships of Neoehrlichia, as well as their reservoir and impact on wildlife, pets and humans.

Hepatozoon silvestris sp. nov.: morphological and molecular characterization of a new species of Hepatozoon (Adeleorina: Hepatozoidae) from the European wild cat (Felis silvestris silvestris).

SUMMARY Based on morphological and genetic characteristics, we describe a new species of Hepatozoon in the European wild cat (Felis silvestris silvestris), herein named Hepatozoon silvestris sp. nov. The study also provides the first data on the occurrence of H. felis in this wild felid. Hepatozoon meronts were observed in multiple cross-sections of different organs of four (44%) cats. Additionally, extracellular forms, resembling mature gamonts of Hepatozoon, were found in the spleen and myocardium of two cats. Furthermore, tissues of six animals (67%) were positive by PCR. Hepatozoon felis was identified infecting one cat (11%), whereas the 18S rRNA sequences of the remaining five cats (56%) were identical, but distinct from the sequences of H. felis. Phylogenetic analyses revealed that those sequences form a highly supported clade distant from other Hepatozoon spp. Future studies should include domestic cats from the areas where the wild cats positive for H. silvestris sp. nov. were found, in order to investigate their potential role to serve as intermediate hosts of this newly described species. Identification of its definitive host(s) and experimental transmission studies are required for elucidating the full life cycle of this parasite and the possible alternative routes of its transmission.

Molecular analysis of Ixodes rugicollis, Candidatus Neoehrlichia sp. (FU98) and a novel Babesia genotype from a European badger (Meles meles).

The European badger (Meles meles) is a widespread mammal in most countries of the European continent, with increasingly recognized veterinary/medical importance owing to its preferred habitats (including pastures and urban environments), broad spectrum of food items, and role as a game hunting target. However, ticks and tick-borne pathogens associated with badgers are only partly known, and most of them have not yet been analysed with molecular biological methods The aim of this study was to perform molecular taxonomic analysis of ticks collected from a road-killed European badger, as well as to molecularly investigate its ticks and blood sample for the presence of Anaplasmataceae and piroplasms. Ticks from the badger were morphologically identified as females of Ixodes rugicollis. Based on its cytochrome oxidase subunit I (COI) and 16S rRNA sequences, I. rugicollis phylogenetically clustered together with I. lividus and I. arboricola, i.e. other members of the subgenus Pholeoixodes. The blood sample of the badger contained the DNA of Candidatus Neoehrlichia sp. (FU98) recently identified in red fox in Austria and the Czech Republic. This genotype is most closely related to Ca. N. lotoris (from raccoons in North America), and has lower sequence identity with the I. ricinus-transmitted zoonotic agent, Ca. N. mikurensis found in Eurasia. In the blood of the badger and in one female I. rugicollis, the DNA of a new Babesia genotype was also present, which differed from a piroplasm detected in M. meles in Spain, and clustered phylogenetically in the B. microti clade. Phylogenetic analysis of I. rugicollis (based on two genetic markers) confirms its status in subgenus Pholeoixodes. Ca. Neoehrlichia sp. (FU98) was identified for the first time in M. meles and in Hungary. In addition, a molecularly previously not yet characterized Babesia genotype occurs in badgers in Central Europe.

Dermacentor reticulatus, a putative vector of Babesia cf. microti (syn. Theileria annae) piroplasm

Babesia cf. microti (syn. Theileria annae, Babesia microti-like, Babesia vulpes) is a recently recognized tick-borne piroplasm that infects domestic and wild carnivores. Although Ixodes hexagonus is considered as the leading candidate responsible for the transmission, its capacity to act as a competent vector has not yet been confirmed. This study reports the occurrence of B.cf. microti in unfed Dermacentor reticulatus for the first time, suggesting that this tick species may be implicated in the life cycle of this canine parasite. Out of 128 questing D. reticulatus ticks collected in eastern Austria, nine (7%) and four (3%) of them were found to be PCR positive for B. canis and B. cf. microti, respectively. Although the data presented here are not sufficient to explicitly state that D. reticulatus is a competent vector of B. cf. microti, our results can at least give a hint for future studies, which need to include experimental transmission in order to confirm its vector competence and possible involvement in the transmission of this babesial species.

Contributions to the phylogeny of Ixodes (Pholeoixodes) canisuga, I. (Ph.) kaiseri, I. (Ph.) hexagonus and a simple pictorial key for the identification of their females

BackgroundIn Europe, hard ticks of the subgenus Pholeoixodes (Ixodidae: Ixodes) are usually associated with burrow-dwelling mammals and terrestrial birds. Reports of Pholeoixodes spp. from carnivores are frequently contradictory, and their identification is not based on key diagnostic characters. Therefore, the aims of the present study were to identify ticks collected from dogs, foxes and badgers in several European countries, and to reassess their systematic status with molecular analyses using two mitochondrial markers.ResultsBetween 2003 and 2017, 144 Pholeoixodes spp. ticks were collected in nine European countries. From accurate descriptions and comparison with type-materials, a simple illustrated identification key was compiled for adult females, by focusing on the shape of the anterior surface of basis capituli. Based on this key, 71 female ticks were identified as I. canisuga, 21 as I. kaiseri and 21 as I. hexagonus. DNA was extracted from these 113 female ticks, and from further 31 specimens. Fragments of two mitochondrial genes, cox1 (cytochrome c oxidase subunit 1) and 16S rRNA, were amplified and sequenced. Ixodes kaiseri had nine unique cox1 haplotypes, which showed 99.2–100% sequence identity, whereas I. canisuga and I. hexagonus had eleven and five cox1 haplotypes, respectively, with 99.5–100% sequence identity. The distribution of cox1 haplotypes reflected a geographical pattern. Pholeoixodes spp. ticks had fewer 16S rRNA haplotypes, with a lower degree of intraspecific divergence (99.5–100% sequence identity) and no geographical clustering. Phylogenetic analyses were in agreement with morphology: I. kaiseri and I. hexagonus (with the similar shape of the anterior surface of basis capituli) were genetically more closely related to each other than to I. canisuga. Phylogenetic analyses also showed that the subgenus Eschatocephalus (bat ticks) clustered within the subgenus Pholeoixodes.ConclusionsA simple, illustrated identification key is provided for female Pholeoixodes ticks of carnivores (including I. hexagonus and I. rugicollis) to prevent future misidentification of these species. It is also shown that I. kaiseri is more widespread in Europe than previously thought. Phylogenetic analyses suggest that the subgenus Pholeoixodes is not monophyletic: either the subgenus Eschatocephalus should be included in Pholeoixodes, or the latter subgenus should be divided, which is a task for future studies.

Troglostrongylus brevior in an Eurasian lynx (Lynx lynx) from Bosnia and Herzegovina

BackgroundIn the past few years the interest of the scientific community on lungworms of the genus Troglostrongylus has grown due to the increased number of unexpected cases of infections with Troglostrongylus brevior in domestic cats from Mediterranean Europe, likely due to a spill-over from wild reservoirs. Thus, there is a merit to increase our knowledge on the occurrence of this parasite in felids from European regions. The present paper describes lung lesions associated with T. brevior infection in the endangered Eurasian lynx (Lynx lynx) from Bosnia and Herzegovina.FindingsThe carcass of an illegally killed 3-year-old male Eurasian lynx was presented for necropsy at the Faculty of Veterinary Medicine of Sarajevo (Bosnia and Herzegovina). Grossly, multiple, multinodular, consolidated and firm, tan to grey areas, occupying the caudal third of caudal lung lobes, were observed. At cut section, the catarrhal fluid was draining from the airways. Larvae of T. brevior were found in tracheal scraping. The histopathological examination revealed multifocal to coalescing areas, centered on bronchi and bronchioles, and expanded alveoli filled with necrotic debris, degenerated inflammatory cells, mostly neutrophils and macrophages, and multiple cross sections of parasite larvae and thin-walled morulated eggs of lungworms. The paraffin-embedded lung samples were molecularly positive for T. brevior.ConclusionThis paper describes the first record of T. brevior in the Eurasian lynx and the associated host lung pathology. Given its pathogenic potential and the lack of data on troglostrongylosis in lynx populations, the occurrence and impact of Troglostrongylus spp. on wildlife health as well as the role of L. lynx as reservoir of infection for other felids, should be further investigated.

The raccoon dog (Nyctereutes procyonoides) and the raccoon (Procyon lotor)—their role and impact of maintaining and transmitting zoonotic diseases in Austria, Central Europe

The neozoan species raccoon dog (Nyctereutes procyonoides) and raccoon (Procyon lotor) are widespread in Europe and potential vectors of many diseases that can threaten human and domestic animal health. Facing a further spread of these species, it is important to know about (i) pathogens imported and/or (ii) pathogens acquired in the new habitat. Thus, we investigated the parasite fauna of wild raccoon dogs and raccoons from Austria, at the edge of their new distribution range. The eight examined raccoons were nearly free of pathogens including Baylisascaris procyonis, and thus assumed to have a low epidemiological impact, so far. Out of ten raccoon dog specimens, we found one from western Austria to be infected with Echinococcus multilocularis and another three from the eastern wetland regions to harbour adults of Alaria alata. Furthermore, we detected Babesia cf. microti in five of eight raccoon dogs all over Austria but none of our samples were tested positive for Trichinella spp. Nevertheless, the raccoon dog seems to be a relevant host, at least for the zoonotic pathogens E. multilocularis and A. alata, and we suggest to further monitor the raccoon dogs parasite fauna.

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.

First report of Rickettsia raoultii in field collected Dermacentor reticulatus ticks from Austria.

In a set of pooled field collected Dermacentor reticulatus ticks, Rickettsia raoultii, the causative agent of Tick-borne lymphadenopathy/Dermacentor-borne necrosis erythema and lymphadenopathy, was found for the first time in Austria. The coordinates of the positive locations for tick and pathogen abundance are given and shown in a map.