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Dirofilaria immitis and D. repens are filarioid nematodes of animals and humans, transmitted by the bite of infected mosquitoes. Domestic and wild canids are a major natural host and reservoir for these parasites. In this study, we designed a duplex real-time PCR protocol targeting the mitochondrial cytochrome c oxidase subunit I (COI) gene, detecting both D. immitis and D. repens using two primer pairs and two Dirofilaria-specific hydrolysable probes. The sensitivity and specificity of the primers and probes were tested in both experimental and naturally infected samples. The detection limits of this assay were evaluated using plasmid DNA from D. immitis and D. repens. No cross-reaction was observed when testing this system against DNA from other filarial nematodes. The detection limit of the real-time PCR system was one copy per reaction mixture containing 5μl of template DNA. Field application of the new duplex real-time assay was conducted in Corsica. The prevalence rate of D. immitis was 21.3% (20/94) in dogs. In a locality where most dogs with Dirofilaria spp. infection were found, D. immitis and D. repens were detected in 5% (20/389) and 1.5% (6/389) of the Aedes albopictus population, respectively. These results suggest that this sensitive assay is a powerful tool for monitoring dirofilariosis in endemic or high risk areas., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Rhipicephalus bursa is one of 79 species of the genus Rhipicephalus in the family of Ixodidae. In this study, we investigated Anaplasmataceae bacteria associated with R. bursa collected after an epizootic outbreak of ovine anaplasmosis. 76 adult ticks, (60 male and 16 female ticks), were removed from sheep in two farms and all identified as R. bursa, all females were partially engorged. We found that 50% of the ticks were positive in the initial Anaplasmataceae qPCR screening. Bacterial species was identified by analyzing the sequences of amplicons of 23S rRNA, groEL and rpoB genes. 22.4% of ticks contained DNA of Anaplasma phagocytophilum and 7.9% the DNA of Anaplasma ovis. Based on 23S rRNA and groEL genes analysis, we found that 19.7% of ticks contained a potentially new species of Ehrlichia. We propose the status of Candidatus for this uncultured species and we provisionally name it Candidatus Ehrlichia urmitei. No Wolbachia were identified. These results show that R. bursa can be a carrier of Anaplasmataceae bacteria., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Ehrlichia canis and Anaplasma platys are bacteria belonging to the Anaplasmataceae family that cause acute, self-limiting and sometimes fatal vector-borne infections in dogs. These bacteria have been reported worldwide and are transmitted mainly by Rhipicephalus sanguineus. Aside from a report on E. canis once in 1935, no other Anaplasmataceae bacteria have been reported in Algeria to date. The aim of this study was to identify the microbial species implicated in ehrlichiosis and anaplasmosis by a molecular epidemiological survey in dogs. The study was carried out in Kabylie, in northeast Algeria. Sampling was performed in 11 municipalities in the province of Tizi Ouzou and 2 municipalities in the province of Béjaïa. Peripheral blood samples from 110 dogs were screened by qPCR, which is capable of identifying most Anaplasmataceae bacteria. Out of 110, a total of 13 samples screened positive (7/110 E. canis and 6/110 A. platys), and two genetic variants of A. platys and one of E. canis were identified. This is the first study to report the presence of A. platys in dogs from Algeria using a molecular investigative method. This survey was conducted in early spring. As tick activity can affect the prevalence of these pathogens in dogs, further investigations are needed to establish the year-round prevalence of these infections., (Copyright © 2015 Elsevier GmbH. All rights reserved.)