Molecular identification of Theileria and Babesia in sheep and goats in the Black Sea Region in Turkey

This study was carried out to investigate presence and distribution of Theileria and Babesia species via microscopic examination and reverse line blotting (RLB) techniques in sheep and goats in the Black Sea region of Turkey. For this purpose, 1,128 blood samples (869 sheep and 259 goats) were collected by active surveillance from sheep and goats in different provinces of various cities in the region in the years 2010 and 2011. Smears were prepared from the blood samples, stained with Giemsa, and examined under the light microscope for Theileria and Babesia piroplasms. The genomic DNAs were extracted from blood samples. The length of 360–430-bp fragment in the variable V4 region of 18S SSU rRNA gene of Theileria and Babesia species was amplified using the gDNAs. The polymerase chain reaction products were hybridized to the membraneconnected species-specific probes. A total of 38 animals (3.37 %) including 34 sheep (3.91 %) and 4 goats (1.54 %) were found to be positive for Theileria spp. piroplasms in microscopic examination of smears while Babesia spp. piroplasm could not detected. Infection rates were 34.64 % in sheep, 10.04 % in goats, and totally 28.99 % for Theileria ovis while 0.58 % in sheep and totally 0.44 % for Babesia ovis. However, Theileria sp. OT3 was detected in 2.65 % of sheep and 2.04 % of all animals; besides Theileria sp., MK had 0.58 % prevalence in sheep and 0.77 % in goats, with a total 0.62 % with RLB. Although T. ovis and Theileria sp. MK were determined in both sheep and goats, B. ovis and Theileria sp. OT3 were observed only in the sheep. These results provide the first detailed molecular data for sheep and goat theileriosis and babesiosis in the region

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

Türkiye’nin Karadeniz Bölgesindeki koyun ve keçilerde kene enfestasyonları

WOS:000303296300004This study was carried out to determine the status of tick infestation of sheep and goats in some cities (Bolu, Kastamonu, corum, Samsun,Tokat, Giresun and Bayburt) in the Black Sea region of Turkiye. For this purpose, a total of 2608 small ruminants (2161 sheep, 447 goat) in 53 town were examined in the period of 2010-2011 and 812 of them (665 sheep, 147 goat) were infested. Whole body parts of the animals were examined and 2797 ticks belong to five genus and 12 species [Rhipicephalus turanicus (28.63%), Haemaaphysalis parva (22.59%), Rhipicephalus bursa (18.26%), Dermacentor marginatus (16.55%), Rhipicephalus sanguineus (3.32%), Ixodes ricinus (2.46%), Haemaphysalis punctata (2.35%), Hyalomma marginatum (2.21%), Haemaphysalis sulcata (1.39%), Hyalomma excavatum (1.17%), Haemaphysalis concinna (0.53%), Hyalomma detritum (0.46%)] were collected. Infestation rates in sheep and goats were 30.77% and 32.88% respectively. It was found that R. turanicus, H. parya and R. bursa were the dominant species on sheep and goats in the region. Seasonal distribution of identified tick species was also determined. Rhipicephalus and Hyalomma species were detected in spring and summer while Haemaphysalis in winter, spring and autumn and Dermacentor and Ixodes in all seasons on sheep and goats in this region.Karadeniz Bölgesi’ndeki bazı illerde (Bolu, Kastamonu, Çorum, Samsun, Tokat, Giresun ve Bayburt) koyun ve keçilerde bulunan kene türleri ile bu türlerin enfestasyon oranlarının ve mevsimsel dağılımlarının belirlenmesi amacıyla yürütülmüştür. Bu amaçla 2010 ve 2011 yıllarında bölgede yer alan 53 farklı yerleşim merkezinden toplam 2608 küçükbaş hayvan (2161 koyun, 447 keçi) kene enfestasyonu yönünden muayene edilmiş ve 812 tanesi (665 koyun, 147 keçi) enfeste bulunmuştur. Hayvanların tüm vücutları kene enfestasyonu yönünden muyene edilmiş, 5 soya ait 12 türde [Rhipicephalus turanicus (%28.63), Haemaphysalis parva (%22.59), Rhipicephalus bursa (%18.26), Dermacentor marginatus (%16.55), Rhipicephalus sanguineus (%3.32), Ixodes ricinus (%2.46), Haemaphysalis punctata (%2.35), Hyalomma marginatum (%2.21), Haemaphysalis sulcata (%1.39), Hyalomma excavatum (%1.17), Haemaphysalis concinna (%0.53), Hyalomma detritum (%0.46)] toplam 2797 adet kene toplanmıştır. Koyun ve keçilerdeki enfestasyon oranları sırasıyla %30.77 ve %32.88 olarak belirlenmiş ve en yaygın türlerin R. turanicus, H. parva ve R. bursa olduğu ortaya konmuştur. Tespit edilen türlerden Rhipicephalus ve Hyalomma’ların ilkbahar ve yaz aylarında görülürken; Haemaphysalis’lerin kış, ilkbahar ve sonbahar; Dermacentor ve Ixodes’lerin ise bütün mevsimlerde hayvanlar üzerinde bulunduğu görülmüştü

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

Molecular identification of Theileria and Babesia in ticks collected from sheep and goats in the Black Sea region of Turkey

WOS:000347158100007PubMed:25260692A molecular survey was undertaken in the Black Sea region of Turkey to determine the presence of Theileria and Babesia species of medical and veterinary importance. The ticks were removed from sheep and goats, pooled according to species and locations, and analyzed by PCR-based reverse line blot (RLB) and sequencing. A total of 2241 ixodid ticks belonging to 5 genus and 12 species were collected and divided into 310 pools. Infection rates were calculated as the maximum likelihood estimation (MLE) with 95 % confidence intervals (CI). Of the 310 pools tested, 46 (14.83 %) were found to be infected with Theileria or Babesia species, and the overall MLE of the infection rate was calculated as 2.27 % (CI 1.67-2.99). The MLE of the infection rates were calculated as 0.691 % (CI 0.171-1.78) in Haemaphysalis parva, 1.47 % (CI 0.081-6.37) in Rhipicephalus sanguineus, 1.84 % (CI 0.101-7.87) in Ixodes ricinus, 2.86 % (CI 1.68-4.48) in Rhipicephalus turanicus, 5.57 % (CI 0.941-16.3) in Hyalomma marginatum, and 6.2 % (CI 4.02-9.02) in Rhipicephalus bursa. Pathogens identified in ticks included Theileria ovis, Babesia ovis, Babesia bigemina, and Babesia microti. Most tick pools were infected with a single pathogen. However, five pools displayed mixed infections with T. ovis and B. ovis. This study provides the first molecular evidence for the presence of B. microti in ticks in Turkey.Scientific and Technical Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [109 O 766]; Commission of Scientific Research Projects, Karamanoglu Mehmetbey UniversityKaramanoglu Mehmetbey University [45-M-12]This work was supported financially by grants (109 O 766) from the Scientific and Technical Research Council of Turkey (TUBITAK) and (45-M-12) Commission of Scientific Research Projects, Karamanoglu Mehmetbey University. We thank all veterinarians, technicians, and also animal breeders in the region for their kind help during sample collection. We are also grateful to Weidong Gu (Health Statistician, PhD, Centers for Disease Control and Prevention, Atlanta, GA, USA) for MLE-IR program used for estimating infection rates of pooled samples