Molecular detection and genetic identification of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in Turkey.
Babesia spp., Theileria spp. and Anaplasma spp. are significant tick-borne pathogens of livestock globally. In this study, we investigated the presence and distribution of Babesia bigemina, Theileria annulata, Theileria orientalis and Anaplasma marginale in cattle from 6 provinces of Turkey using species-specific PCR assays. The PCR were conducted using the primers based on the B. bigemina rhoptry-associated protein 1a (BbiRAP-1a), T. annulata merozoite surface antigen-1 (Tams-1), T. orientalis major piroplasm surface protein (ToMPSP) and A. marginale major surface protein 4 (AmMSP4) genes, respectively. Fragments of B. bigemina internal transcribed spacer (BbiITS), T. annulata internal transcribed spacer (TaITS), ToMPSP and AmMSP4 genes were sequenced for phylogenetic analysis. PCR results revealed that the overall infections of A. marginale, T. annulata, B. bigemina and T. orientalis were 29.1%, 18.9%, 11.2% and 5.6%, respectively. The co-infection of two or three pathogens was detected in 29/196 (15.1%) of the cattle samples. The results of sequence analysis indicated that BbiRAP-1a, BbiITS, Tams-1, ToMPSP and AmMSP4 were conserved among the Turkish samples, with 99.76%, 99-99.8%, 99.34-99.78%, 96.9-99.61% and 99.42-99.71% sequence identity values, respectively. In contrast, the Turkish TaITS gene sequences were relatively diverse with 92.3-96.63% identity values. B. bigemina isolates from Turkey were found in the same clade as the isolates from other countries in phylogenetic analysis. On the other hand, phylogenetic analysis based on T. annulata ITS sequences revealed significant differences in the genotypes of T. annulata isolates from Turkey. Additionally, the T. orientalis isolates from Turkish samples were classified as MPSP type 3 genotype. This is the first report of type 3 MPSP in Turkey. Moreover, AmMSP4 isolates from Turkey were found in the same clade as the isolates from other countries. This study provides important data for understanding the epidemiology of tick-borne diseases and it is expected to improve approach for diagnosis and control of tick-borne diseases in Turkey.
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Mo
M
Zhou
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Shinuo
S
Cao
Faculty of Veterinary Medicine, Selcuk University, Konya, Turkey.
Ferda
F
Sevinc
Faculty of Veterinary Medicine, Selcuk University, Konya, Turkey.
Mutlu
M
Sevinc
Faculty of Veterinary Medicine, Selcuk University, Konya, Turkey.
Onur
O
Ceylan
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Paul Franck Adjou
PFA
Moumouni
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Charoonluk
C
Jirapattharasate
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Mingming
M
Liu
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Guanbo
G
Wang
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Aiko
A
Iguchi
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Patrick
P
Vudriko
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan.
Hiroshi
H
Suzuki
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080-8555, Japan. Electronic address: gen@obihiro.ac.jp.
Xuenan
X
Xuan
D042322:Anaplasma marginale / Q000235:genetics*; D000712:Anaplasmosis / Q000453:epidemiology / Q000382:microbiology*; D000818:Animals; D000953:Antigens, Protozoan / Q000235:genetics / Q000302:isolation & purification; D001403:Babesia / Q000235:genetics*; D001404:Babesiosis / Q000453:epidemiology / Q000469:parasitology*; D002417:Cattle; D002418:Cattle Diseases / Q000453:epidemiology* / Q000382:microbiology / Q000469:parasitology; D004269:DNA, Bacterial / Q000235:genetics; D016054:DNA, Protozoan / Q000235:genetics; D014644:Genetic Variation; D010802:Phylogeny; D016794:Theileria / Q000235:genetics*; D013801:Theileriasis / Q000453:epidemiology / Q000469:parasitology*; D014421:Turkey / Q000453:epidemiology
D000953:Antigens, Protozoan; D004269:DNA, Bacterial; D016054:DNA, Protozoan
Cattle; Species-specific PCR; Tick-borne pathogen; Turkey