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    Molecular detection of tick-borne rickettsial and protozoan pathogens in domestic dogs from Turkey
    (Bmc, 2015) Aktas, Munir; Ozubek, Sezayi; Altay, Kursat; Ipek, Neval Duygu Sayin; Balkaya, Ibrahim; Utuk, Armagan Erdem; Kirbas, Akin
    Background: Canine tick-borne parasites have emerged in recent years, showing a wider geographic distribution and increased global prevalence. In addition to their veterinary importance, domestic dogs play an important role in the transmission cycles of some agents by acting as reservoirs and sentinels. This study investigated Babesia, Theileria, Anaplasma, and Ehrlichia species in asymptomatic dogs in ten provinces of Turkey. Methods: DNA obtained from blood samples collected from 757 domestic dogs (243 stray, 351 shelter, 163 pet) of both sexes and various ages were evaluated using PCR and reverse line blotting (RLB) assays. Results: Of the 757 dogs tested, 41 (5.4%) were found to be infected with one or more parasites. Ehrlichia canis (37/757, 4.9%) was the most common canine tick-borne pathogen, followed by Anaplasma platys (4/757, 0.5%). Babesia canis and Theileria annulata were each detected in 1 (0.13%) sample. Combined infection of E. canis and A. platys was detected in 2 (0.3%) samples. The prevalence of tick-borne pathogens was higher in adult dogs (6.8%) than in those under one year old (3.1%). Difference in infection rate of male and female dogs was not significant. Pet dogs had a lower prevalence of infection (1.2%) compared to stray (7.4%) and shelter dogs (6%) although the difference between stray and shelter dogs was not significant. Conclusions: Babesia canis, T. annulata, A. platys, and E. canis species were identified at the molecular level in dogs in several provinces of Turkey, with E. canis being the most common species among tick-borne pathogens. Detailed studies should be conducted regarding the existence and prevalence of B. canis and Dermacentor reticulatus in eastern Turkey.
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    Molecular Evidence for Transstadial Transmission of Ehrlichia canis by Rhipicephalus sanguineus sensu lato Under Field Conditions
    (Oxford Univ Press Inc, 2018) Ipek, Neval Duygu Sayin; Ozubek, Sezayi; Aktas, Munir
    This study investigated possible transstadial transmission of Ehrlichia canis by Rhipicephalus sanguineus sensu lato collected from shelter dogs and the shelter grounds in Diyarbakir Province of south-eastern Turkey. Totally 225 engorged nymphs were collected from eight infected dogs with E. canis and incubated at 28 degrees C for moulting. Unfed ticks from the shelter grounds comprising 1,800 larvae, 3,100 nymphs, and 85 adults were sorted according to sampling origin, life stage, and sex into 116 pools and screened by 16S rRNA PCR. Nine out of 26 pools of unfed adult ticks were positive for E. canis, with overall infection rate maximum likelihood estimation (MLE) of 4.83 (CI 2.39-8.87). E. canis was detected in three of 12 male pools (MLE 3.22, CI 0.86-8.83) and six of 14 female pools (MLE 6.16, CI 2.59-12.90). No adult pools collected from the shelter grounds were positive. Among 62 unfed nymph pools collected from the shelter, six were infected with E. canis (MLE 0.20, CI 0.08-0.42). No E. canis DNA was detected in any of the larva pools. Our results revealed molecular evidence for transstadial transmission of E. canis by R. sanguineus s.l. both from larva to nymph and from nymph to adult. We found no evidence of transovarial transmission.
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    Molecular investigations of Hepatozoon species in dogs and developmental stages of Rhipicephalus sanguineus
    (Springer, 2013) Aktas, Munir; Ozubek, Sezayi; Ipek, Duygu Neval Sayin
    The occurrence and distribution of Hepatozoon species in stray dogs, and the developmental stages of Rhipicephalus sanguineus detached from the same dogs in DiyarbakA +/- r Province, Turkey is reported. A total of 328 ticks, including 133 adults (55 males and 75 females consist of 63 partially engorged and 15 fully engorged) and 195 nymphs (91 partially engorged and 104 fully engorged) were detached from the dogs. Fully engorged nymphs and females were incubated at 27 A degrees C and relative humidity of 85 % to molt to adult stage and recover eggs. The ticks were pooled according to sex and developmental stage. No Hepatozoon gamonts were found, whereas, by PCR, 15.87 % (10/63) of the dogs were infected with Hepatozoon canis. Of the 68 tick pools tested, 14 (20.58 %) pools were infected with Hepatozoon spp., an overall maximum likelihood estimation of prevalence of 4.9 % (95 % confidence intervals (CI) = 2.85-7.93 %) per 100 ticks. Maximum likelihood estimation of the infection rate varied by tick sex and developmental categories, ranging from 1.75 % (95 % CI = 0.11-8.11 %) in fed males to 6.81 % (95 % CI = 2.07-17.46 %) in unfed females. One amplicon from a fed adult female was 99 % identical to the sequence for Hepatozoon felis. The remaining sequences isolated from both dogs and ticks shared 99-100 % similarity with the corresponding H. canis isolates. This is the first detection of H. canis and H. felis in the tick R. sanguineus in Turkey.
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    A Molecular Survey of Rickettsias in Shelter Dogs and Distribution of Rhipicephalus sanguineus (Acari: Ixodidae) sensu lato in Southeast Turkey
    (Oxford Univ Press Inc, 2018) Ozubek, Sezayi; Ipek, Duygu Neval Sayin; Aktas, Munir
    Canine tick-borne pathogens are the source of emerging diseases and have important zoonotic relevance. Dogs play a major role in the transmission of several zoonotic tick-borne pathogens, as reservoirs and/or sentinels. To simultaneously detect Anaplasma and Ehrlichia species, a reverse line blot assay was conducted on 219 blood samples collected from autochthonous asymptomatic shelter dogs. One hundred and three (47.0%, CI 40.3-53.9) dogs were positive for one or both rickettsial pathogens. Seventy-one (32.4%, CI 26.3-39.0) dogs were infected with Anaplasma platys and 23 (10.5%, CI 6.8-15.3) with Ehrlichia canis. Concurrent infection with A. platys and E. canis was detected in nine (4.1%, CI 1.9-7.6) dogs. Partial sequences of the 16S ribosomal RNA gene shared 100% identity with the corresponding published sequences for A. platys and E. canis. Infection with Anaplasma phagocytophilum was not detected in the examined dogs. In total, 1018 (range 1-70, mean intensity 13.1, mean abundance 4.6) Rhipicephalus sanguineus (Latreille) (Acari: Ixodidae) sensu lato ticks (45.7% nymphs, 54.3% adults) were collected from the dogs. There was no significant association between Anaplasma/Ehrlichia infection and dog sex or age, but a significant correlation was found between rickettsia infection and presence of R. sanguineus. Improved tick control strategies to reduce the risk of these pathogens spreading among dogs and humans are needed in the region.