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First molecular evidence of Hepatozoon canis infection in red foxes and golden jackals from Hungary.

ABSTRACT: BACKGROUND: Recently, Hepatozoon canis infection has been detected among shepherd, hunting and stray dogs in the southern part of Hungary, which is considered to be free of Rhipicephalus sanguineus sensu lato and close to the border with Croatia. The aim of this study was to acquire information on the possibility that red foxes and/or golden jackals could play a role in the appearance and spread of H. canis in Hungary. METHODS: A conventional PCR was used to amplify a 666 bp long fragment of the Hepatozoon 18S rRNA gene from blood samples collected from 334 foxes shot in 231 locations in 16 counties and 15 golden jackals shot in 9 locations in two southwestern counties close to Croatia. A second PCR assay was performed in some of the samples positive by the first PCR to amplify a larger segment (approximately 1500 bp) of the 18S rRNA gene of Hepatozoon spp. for further phylogenetic analysis. RESULTS: Hepatozoon infection was detected in canids shot in 30 locations and 9 counties. Altogether 26 foxes (8.0%, 95% CI: 5-11%) and 9 jackals (60%, 95% CI: 33-81%) were PCR positive. Hepatozoon canis sequences were obtained from 12 foxes and 7 jackals. DNA sequences from 16 animals were 99-100% similar to H. canis from Croatian foxes or dogs while two of the sequences were 99% similar to an Italian fox. Half (13/26) of the infected red foxes and all golden jackals were shot in the two southwestern counties. CONCLUSIONS: This is the first report on molecular evidence of H. canis in red foxes (Vulpes vulpes) and golden jackals (Canis aureus) from Hungary, which is considered free from the tick vector of H. canis, R. sanguineus. Although no R. sanguineus sensu lato had been found on infected or non-infected wild canids, the detection of authochnous canine hepatozoonosis in Hungary might imply that the range of R. sanguineus sensu lato has reached this country.

SUBMITTER: Farkas R

PROVIDER: S-EPMC4086283 | biostudies-literature | 2014

REPOSITORIES: biostudies-literature

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First molecular evidence of Hepatozoon canis infection in red foxes and golden jackals from Hungary.

Farkas Róbert R Solymosi Norbert N Takács Nóra N Hornyák Ákos Á Hornok Sándor S Nachum-Biala Yaarit Y Baneth Gad G

Parasites & vectors 20140702

<h4>Background</h4>Recently, Hepatozoon canis infection has been detected among shepherd, hunting and stray dogs in the southern part of Hungary, which is considered to be free of Rhipicephalus sanguineus sensu lato and close to the border with Croatia. The aim of this study was to acquire information on the possibility that red foxes and/or golden jackals could play a role in the appearance and spread of H. canis in Hungary.<h4>Methods</h4>A conventional PCR was used to amplify a 666 bp long fr ...[more]

PMID: 24985073

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Project description:BACKGROUND: Hepatozoon canis is a protozoan tick-borne pathogen of dogs and wild canids. Hepatozoon spp. have been reported to infect foxes in different continents and recent studies have mostly used the polymerase chain reaction (PCR) for the detection and characterization of the infecting species. Surveying red foxes (Vulpes vulpes) may contribute to better understanding the epidemiology of canine vector-borne diseases, including hepatozoonosis caused by H. canis in domestic dogs. The present study investigated the prevalence of Hepatozoon spp. by means of histopathology and molecular analysis of different tissues in red foxes from different parts of Portugal. METHODS: Blood and tissues including bone marrow, heart, hind leg muscle, jejunum, kidney, liver, lung, popliteal or axillary lymph nodes, spleen and/or tongue were collected from 91 red foxes from eight districts in northern, central and southern Portugal. Tissues were formalin-fixed, paraffin-embedded, cut and stained with hematoxylin and eosin. Polymerase chain reaction (PCR) amplified a ~650 bp fragment of the 18S rRNA gene of Hepatozoon spp. and the DNA products were sequenced. RESULTS: Hepatozoon canis was detected in 68 out of 90 foxes (75.6%) from all the sampled areas by PCR and sequencing. Histopathology revealed H. canis meronts similar in shape to those found in dogs in the bone marrow of 11 (23.4%) and in the spleen of two (4.3%) out of 47 foxes (p = 0.007). All the 11 foxes found positive by histopathology were also positive by PCR of bone marrow and/or blood. Positivity by PCR (83.0%) was significantly higher (p < 0.001) than by histopathological examination (23.4%) in paired bone marrow samples from the same 47 foxes. Sequences of the 18S rRNA gene of H. canis were 98-99% identical to those in GenBank. CONCLUSIONS: Hepatozoon canis was found to be highly prevalent in red fox populations from northern, central and southern Portugal. Detection of the parasite by histopathology was significantly less sensitive than by PCR. Red foxes are a presumptive reservoir of H. canis infection for domestic dogs.

Project description:BACKGROUND:Domestic dogs are not native to sub-Saharan Africa, which may account for their susceptibility to Babesia rossi, of which endemic black-backed jackals (Canis mesomelas) are natural reservoirs. There is virtually no information on the occurrence of potentially pathogenic haemogregarines (e.g. Hepatozoon canis) or even rickettsial bacteria (e.g. Ehrlichia spp. and Anaplasma spp.) in indigenous canids in sub-Saharan Africa. Such organisms could pose a risk to domestic dogs, as well as to populations of endangered indigenous canid species. RESULTS:Genomic DNA extracted from blood samples taken from 126 free-ranging and 16 captive black-backed jackals was subjected to reverse line blot (RLB) hybridization assay; 82 (57.8%) specimens reacted only with the Ehrlichia/Anaplasma genera-specific probe. Full-length bacterial 16S rRNA gene of five of these specimens was cloned and the recombinants sequenced. The ten 16S rDNA sequences obtained were most closely related, with approximately 99% identity, to Anaplasma sp. South African Dog, various uncultured Anaplasma spp., as well as various Anaplasma phagocytophilum genotypes. Ninety-one specimens were screened for haemogregarines through PCR amplification using the 18S rRNA gene; 20 (21.9%) specimens reacted positively, of which 14 (15.4%) were confirmed positive for Hepatozoon genotypes from within H. canis. Two (2.2%) specimens were found positive for two different Hepatozoon genotypes. CONCLUSIONS:Sequence analyses confirmed the presence of 16S rDNA sequences closely related to A. phagocytophilum and Anaplasma sp. South African Dog as well as two H. canis genotypes in both free-ranging and captive black-backed jackals. Distinguishing between closely related lineages may provide insight into differences in pathogenicity and virulence of various Anaplasma and H. canis genotypes. By building up a more comprehensive understanding of the range and diversity of the bacteria and eukaryotic organisms (piroplasms and haemogregarines) in the blood of indigenous canids, we may gain insight to such infections in these often-endangered species and the potential for horizontal transmission to and from domestic dogs via ticks where favourable conditions exist.

Project description:Golden jackal (Canis aureus) expansion in the last decades has triggered research interest in Europe. However, jackal phylogeny and taxonomy are still controversial. Morphometric studies in Europe found differences between Dalmatian and the other European jackals. Recent genetic studies revealed that African and Eurasian golden jackals are distinct species. Moreover, large Canis aureus lupaster may be a cryptic subspecies of the African golden jackal. Although genetic studies suggest changes in Canis aureus taxonomy, morphological and morphometric studies are still needed. The present study proposes the first comprehensive analysis on a wide scale of golden jackal skull morphometry. Extensive morphometric data of jackal skulls from Europe (including a very large Bulgarian sample), Asia Minor, and North Africa were analysed, by applying recently developed statistical tools, to address the following questions: (i) is there geographic variation in skull size and shape among populations from Europe, Anatolia and the Caucasus?, (ii) is the jackal population from the Dalmatian coast different?, and (iii) is there a clear distinction between the Eurasian golden jackal (Canis aureus) and the African wolf (Canis lupaster sensu lato), and among populations of African wolves as well? Principal component analysis and linear discriminant analysis were applied on the standardized and log-transformed ratios of the original measurements to clearly separate specimens by shape and size. The results suggest that jackals from Europe, Anatolia and the Caucasus belong to one subspecies: Canis aureus moreotica (I. Geoffroy Saint-Hilaire, 1835), despite the differences in shape of Dalmatian specimens. The present study confirmed morphometrically that all jackals included so far in the taxon Canis aureus sensu lato may represent three taxa and supports the hypothesis that at least two different taxa (species?) of Canis occur in North Africa, indicating the need for further genetic, morphological, behavioural and ecological research to resolve the taxonomic uncertainty. The results are consistent with recent genetic and morphological studies and give further insights on golden jackal taxonomy. Understanding the species phylogeny and taxonomy is crucial for the conservation and management of the expanding golden jackal population in Europe.

Dataset Information

First molecular evidence of Hepatozoon canis infection in red foxes and golden jackals from Hungary.

Publications

First molecular evidence of Hepatozoon canis infection in red foxes and golden jackals from Hungary.

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