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IntroductionTick-borne pathogens (TBP) are an important group of organisms that can affect animals and humans all over the world. Equine piroplasmosis (EP), caused by Theileria equi and Babesia caballi, is considered one of the most important tick-borne diseases and can cause significant clinical symptoms and mortality in horses. Moreover, EP plays a restrictive role in international horse traditions and transportation. Although these species can cause similar symptoms, there are different 18S rRNA genotypes of T. equi (five genotypes) and B. caballi (three genotypes). Besides piroplasma species, Anaplasma and hemotropic mycoplasmas (HM) are known as other important tick-borne pathogens reported in horses.MethodsIn this study, we investigated the presence, prevalence, genetic diversity, and phylogenetic analyses of TBPs using PCRs and DNA sequencing in grazing horses in Kyrgyzstan. For these purposes, a total of 311 blood samples were collected from Chuy, Issyk-Kul, Naryn, Osh, Talas, and Jalal-Abad.ResultsDNA amplification of TBP revealed that 23 (7.40%) out of 311 samples were found to be positive for T. equi. However, B. caballi, HM, A. phagocytophilum, and A. capra were not detected in this study. The infection rate of T. equi was higher in males (8.11%) than in females (6.35%) (p=0.2880) and in those older than 5 years (9.02%) than in the 1-4 age group (6.35%) (p=0.1950). Phylogenetic analysis of 18S rRNA revealed that A and E genotypes of T. equi have circulated in grazing horses in Kyrgyzstan.DiscussionInformation about the genetic diversity of T. equi is important for understanding the population dynamics of the species and developing effective control strategies against this pathogen. This is the first molecular investigation of A. capra in horses in Kyrgyzstan. Although this pathogen has been detected in different hosts in Kyrgyzstan, it was not detected in this study. However, considering the wide host spectrum of A. capra, it is thought that more large-scale studies are needed to understand the effect of horses on the epidemiology of this pathogen.

Tick-borne parasitic diseases cause mild to severe infections among vertebrate hosts, including dogs. Species in the genus Babesia are important tick-borne pathogens and have worldwide distributions. Although there are data on the prevalence and distribution of Babesia species among dogs around the world, there is no information available in Kyrgyzstan, according to a literature review. In this study, 337 dogs were screened by nested PCR for the presence of the 18S small subunit ribosomal RNA (18S SSU rRNA) gene of piroplasm species. Overall prevalence was 6.23% (21/337) for Babesia/Theileria spp. DNA sequencing of positively tested samples revealed that eighteen samples were infected with Babesia vogeli (B. vogeli) (5.34%), two samples with B. vulpes (0.59%), and one sample with Theileria ovis (T. ovis) (0.29%). The phylogenetic analyses and nucleotide sequences in contrast with those present in GenBank revealed that two nucleotide substitutions (594th and 627th) were found between B. vogeli isolates, including ours, indicating that the mutation is relatively rare. The sequences of other pathogens obtained in this study confirmed 100% nucleotide identity with B. vulpes and T. ovis sequences in GenBank. To the best of our knowledge, B. vogeli, B. vulpes, and T. ovis were detected for the first time in dogs from Kyrgyzstan, and it is thought that results will contribute to the understanding of the epidemiology of canine tick-borne pathogens in the country.

Mycoplasma haemocanis (Mhc) and Candidatus Mycoplasma haematoparvum (CMhp) have been investigated using species specific PCR and sequencing in 170 dogs from Kyrgyzstan. Maximum likelihood estimation (MLE) of the infection rates with 95% confidence intervals (CI) was calculated. The molecular prevalence of hemoplasma infection was 5.29% (CI 2.57-9.34). It was found that, five (2.94%, CI 1.06-6.22) samples were found to be infected with Mhc, one (0.59%, CI 0.03-2.57) sample with CMhp and three (1.76%, CI 0.44-4.52) samples with both species. These results demonstrate that dogs can be exposed to each haemoplasma species and provide first molecular evidence for these species in Kyrgyzstan.

Canine hepatozoonosis is a tick-borne protozoan disease and widespread in Europe, Africa, Asia and America. There is not any available data about the presence of Hepatozoon infections in dogs in Kyrgyzstan. In the study we aimed that investigate the presence of Hepatozoon canis and the prevalance of Hepatozoon infections in dogs from Kyrgyzstan using polymerase chain reaction (PCR) and sequence analysis. To determine the prevelance of hepatozoonosis in dogs, a total of 170 blood samples were applied to PCR to amplify a fragment of 666 bp found in 18S SSU rRNA gene of Hepatozoon spp. The PCR results shown that Hepatozoon infection rate was 28.8% (49/170) in dogs. Eleven representative positive samples were sequenced to classification of the species. The nucleotide sequences were compared to the H. canis sequences which registered in GenBank using the basic local alignment search tool. Results of sequence analyse of 11 amplicons indicated that 8 were 100% identical and the other 3 sequences shared 99% similarity with H. canis. The sequences were deposited in Genbank with accession numbers from MG917709 to MG917719. It was the first record of H. canis in dogs in Kyrgyzstan.

Objective:This study aimed to investigate Anaplasma phagocytophilum and related strains (A. phagocytophilum-like 1 and like 2) in sheep and goats for the first time in Sivas province with molecular techniques.Methods:The study material was composed of 247 animal (159 sheep and 88 goats) blood samples from four districts of Sivas province (Sivas City Center, Kangal, Koyulhisar, and Yıldızeli). A. phagocytophilum and related strains were screened with polymerase chain reaction (PCR), PCR-RFLP, and DNA sequence analysis.Results:A. phagocytophilum related strains were found in 44.93% (111/247) of the small ruminants using PCR. The infection rate was 45.91% (73/159) in sheep and 43.18% (38/88) in goats. In this study, 110 samples were positive for only A. phagocytophilum-like 1, while A. phagocytophilum-like 1 and like 2 were mix-infection in one sample. A. phagocytophilum was not detected in sheep or goats. Two randomly selected PCR products were sequenced in both directions, and the consensus sequences were deposited on the GenBank under accession numbers: ON598644 and ON598645. Nucleotide similarity of 99.34-100% was determined between A. phagocytophilum-like 1 isolates obtained in this study and those of A. phagocytophilum-like 1 isolates present in the GenBank database.Conclusion:This study provides the first molecular data on A. phagocytophilum-like 1 and like 2 in Sivas province. Considering the high positive rate of the A. phagocytophilum-like 1 in sheep and goats, there is a paucity of data on clinical symptoms and vector species of the pathogen. Therefore, further studies are needed to investigate the vector tick species and clinical symptoms of the pathogen in the host.

Abstract  A polymerase chain reaction (PCR) assay followed by partial sequencing of the 16S ribosomal RNA gene was performed for the presence of Ehrlichia and/or Anaplasma. A total of 242 ixodid ticks were collected from domestic ruminants and their shelters, as well as humans, and their individual salivary glands were dissected out for DNA. From the 242 ticks analyzed, six (2.47%), comprising three Hyalomma anatolicum anatolicum, one Rhipicephalus bursa, and two Rhipicephalus sanguineus, were positive. Of these sequenced samples directly obtained from the PCR products, three sequences from H. a. anatolicum were identical to that of the gene of Ehrlichia spp. strains. One sequence identified in R. bursa was closely related to Anaplasma platys. The remaining two sequences detected in R. sanguineus were similar to that of the gene of Anaplasma ovis. The study presented here provides preliminary data regarding the presence of rickettsial pathogens in ticks in Turkey. [ABSTRACT FROM AUTHOR]

Abstract   Theileria sp. MK in sheep and goats were detected first time by polymerase chain reaction (PCR) and detection limit of PCR and reverse line blotting (RLB) were compared. A part of 18S ssu rRNA gene was amplified from blood samples that were taken from sheep and goats naturally infected with Theileria sp. MK by PCR. Detection limit of both PCR and RLB methods was one infected cell in 107 sheep erythrocytes. Nine hundred twenty field samples that had been tested previously by RLB were evaluated by the PCR assay. As found by RLB previously, 12 of 920 (1.30%) samples were detected as positive by PCR. Two positive PCR products, one of which was from sheep and the other from goat, were sequenced. These sequences were identical to the reported nucleotide sequence of Theileria sp. MK. It is concluded that the PCR described in this study will be useful for epidemiological studies and for discrimination between Theileria sp. MK and other Theileria species. In addition, PCR has superiority over RLB because of its ease of use and time period required. [ABSTRACT FROM AUTHOR]

Abstract??In this study, PCR and thin blood smear-based diagnostic methods were used to assess the frequency ofBabesiainfection in small ruminants. A total of 300 sheep and 100 goats from 37 randomly selected herds located in eight locations of eastern Turkey were examined for the presence ofBabesiainfection and any tick species on the body of the animals. Of 400 blood samples examined, 6 (1.5%) were positive forBabesiaspp. piroplasms upon microscopic examination, whereas 33 (8.25%) were positive for the presence ofB. ovisby PCR. The prevalence of babesiosis in small ruminants detected by PCR was significantly higher than obtained in microscopic examination of thin blood smears (P<0.05). Thirty-three animals produced the DNA fragment specific forBabesia ovisof which 32 (10.66%) were sheep and 1 (1%) was goat. The difference between the prevalence ofBabesiainfection in sheep and goats were statistically significant (P<0.05). The prevalence ofBabesiainfection in different age groups of sheep were statistically non-significant (P?>?0.05). The frequency ofB. ovisinfection was higher in herds with tick burden than no tick burden (P<0.05). Seven amplicons (six from sheep and one from goat) were sequenced. The resulting sequences were identical to the recently reported nucleotide sequence ofB. ovis. A total of 510 ticks belonging to theRhipicephalusspp. were collected from sheep. Ticks were identified to beR. bursaandR. turanicuson the basis of morphological features. [ABSTRACT FROM AUTHOR]

Abstract 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.