Anaplasma marginale msp1a geninin yeni bir tandem raporunu da içeren, Türkiye’nin Karaman yöresindeki sığırlarda Anaplasma ve Ehrlichia türlerinin moleküler araştırması

WOS:000472627700005Tick-borne pathogens cause serious health problems and loss of productivity in domesticated and wild animals. A. molecular study was performed to detect the frequency of infection with Anaplasma/Ehrtichia (A/E) in cattle from Karaman province of Turkey. Venous blood samples were taken from 150 apparently healthy cattle in 2016. After amplification the hypervariable VI region of the 16S rRNA gene of A/E species, a reverse line blot (RLB) assay was performed using species-specific probes. Since some samples gave signal only to A/E catch-all probe, the samples analyzed in terms of major surface proteins (MSPs) of Anaplasma marginale. Genetic diversity and tandem repeat analysis were made for msp1 alpha gene sequences of A. marginale.Anaplasma-like bodies were detected in four (2.66%) animals via microscopic examination. Anaplasma centrale was detected in eight (5.33%) animals via RLB. When the samples were examined in terms of A. marginale msp 1 a gene with semi-nested PCR, a total of nine (6.00%) animals [six of them (4.00%) were positive for A. centrale with RLB] were found to be infected with A. marginale. In addition, the sequences of MSP 1a amplicons revealed one new tandem repeat (Tr70). According to these results, it was determined that A. marginale and A. centrale were found in cattle in Karaman province and this study provided the first evidence of genetic diversity of A. marginale with one new tandem repeat in cattle in the region.Kene kaynaklı patojenler evcil ve yabani hayvanlarda ciddi sağlık problemlerine ve verim kaybına neden olur. Karaman ilindeki sığırlarda Anaplasma / Ehrlichia (A/E) ile enfeksiyon sıklığını saptamak için moleküler bir çalışma yapıldı. Venöz kan numuneleri görünüşte sağlıklı olan 150 sığırdan 2016 yılında alınmıştır. A/E türlerinin 16S rRNA geninin değişken V1 bölgesi amplifiye edildikten sonra tür spesifik problar kullanılarak reverse line blot (RLB) deneyi gerçekleştirilmiştir. Bazı örnekler sadece A/E probuna sinyal verdiğinden, örnekler Anaplasma marginale'nin major surface proteinleri (MSPs) açısından analiz edilmiştir. A. marginale'nin msp1α gen dizileri için genetik çeşitlilik ve tandem tekrar analizi yapıldı. Mikroskobik inceleme ile dört (%2.66) hayvanda Anaplasma benzeri cisimler tespit edildi. Anaplasma centrale RLB ile sekiz (%5.33) hayvanda tespit edildi. Örnekler seminested PZR ile A. marginale msp1a geni açısından incelediğinde, toplam dokuz (%6.00) hayvanın [6'sı (%4.00) RLB ile A. centrale açısından pozitif olan] A. marginale ile enfekte olduğu bulunmuştur. Ek olarak, MSP1a amplikonlarının dizileri bir tane yeni tandem tekrarı (Tr70) ortaya çıkardı. Bu sonuçlara göre Karaman ilindeki sığırlarda A. marginale ve A. centrale'nin bulunduğu tespit edilmiş ve bu çalışma bölgedeki sığırlarda bir yeni tandem tekrarı ile A. marginale’nin genetik çeşitliliğinin ilk kanıtını sağlamıştır.Commission of Scientific Research Projects, Karamanoglu Mehmetbey UniversityKaramanoglu Mehmetbey University [41-M-16]This work was supported financially by a grant (41-M-16) from the Commission of Scientific Research Projects, Karamanoglu Mehmetbey University

Molecular Detection and Phylogeny of Anaplasma phagocytophilum and Related Variants in Small Ruminants from Turkey

Anaplasma phagocytophilum causes tick-borne fever in small ruminants. Recently, novel Anaplasma variants related to A. phagocytophilum have been reported in ruminants from Tunisia, Italy, South Korea, Japan, and China. Based on 16S rRNA and groEL genes and sequencing, we screened the frequency of A. phagocytophilum and related variants in 433 apparently healthy small ruminants in Turkey. Anaplasma spp. overall infection rates were 27.9% (121/433 analyzed samples). The frequency of A. phagocytophilum and A. phagocytophilum-like 1 infections was 1.4% and 26.5%, respectively. No A. phagocytophilum-like 2 was detected in the tested animals. The prevalence of Anaplasma spp. was comparable in species, and no significant difference was detected between sheep and goats, whereas the prevalence significantly increased with tick infestation. Sequencing confirmed PCR-RFLP data and showed the presence of A. phagocytophilum and A. phagocytophilum-like-1 variant in the sampled animals. Phylogeny-based on 16S rRNA gene revealed the A. phagocytophilum-like 1 in a separate clade together with the previous isolates detected in small ruminants and ticks. In this work, A. phagocytophilum-like 1 has been detected for the first time in sheep and goats from Turkey. This finding revealed that the variant should be considered in the diagnosis of caprine and ovine anaplasmosis

Bovine Babesiosis in Turkey: Impact, Current Gaps, and Opportunities for Intervention

Bovine babesiosis is a global tick-borne disease that causes important cattle losses and has potential zoonotic implications. The impact of bovine babesiosis in Turkey remains poorly characterized, but several Babesia spp., including B. bovis, B. bigemina, and B. divergens, among others and competent tick vectors, except Rhipicephalus microplus, have been recently identified in the country. Bovine babesiosis has been reported in all provinces but is more prevalent in central and highly humid areas in low and medium altitude regions of the country housing approximately 70% of the cattle population. Current control measures include acaricides and babesicidal drugs, but not live vaccines. Despite the perceived relevant impact of bovine babesiosis in Turkey, basic research programs focused on developing in vitro cultures of parasites, point-of-care diagnostic methods, vaccine development, “omics” analysis, and gene manipulation techniques of local Babesia strains are scarce. Additionally, no effective and coordinated control efforts managed by a central animal health authority have been established to date. Development of state-of-the-art research programs in bovine babesiosis to address current gaps in knowledge and implementation of long-term plans to control the disease will surely result in important economic, nutritional, and public health benefits for the country and the region

Molecular detection of tick-borne rickettsial and protozoan pathogens in domestic dogs from Turkey

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