First Molecular Characterization of Anaplasma marginale in Cattle and Rhipicephalus (Boophilus) microplus Ticks in Cebu, Philippines

Journal of Veterinary Medical Science

Rochelle Haidee Daclan YBAÑEZ¹  Hassan HAKIMI⁴  Kotaro MATSUMOTO²  Zandro Obligado PEREZ³  Naoaki YOKOYAMA⁴  Shin-ichiro KAWAZU¹  Shirleny Reyes GABOTERO³  Thillaiampalam SIVAKUMAR¹  Jowarren Catingan RATILLA⁵  Hisashi INOKUMA⁴  Adrian Patalinghug YBAÑEZ² 

[1] National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080–8555, Japan;Department of Veterinary Clinical Sciences, Obihiro University of Agriculture and Veterinary Medicine, Obihiro, Hokkaido 080–8555, Japan;GPY Veterinare Animale - Group of Veterinary Clinics, Cebu City 6000, Philippines;United Graduate School of Veterinary Sciences, Gifu University, Gifu 501–1193, Japan;National Dairy Authority, Cebu City 6000, Philippines

关键词: Anaplasma marginale;    cattle;    PCR;    Philippines;    Rhipicephalus (Boophilus) microplus;   

DOI  :  10.1292/jvms.12-0268

学科分类：兽医学

来源: Japanese Society of Veterinary Science

【 摘 要 】

References(33)Cited-By(11)Anaplasma marginale has been detected in the Philippines only by peripheral blood smear examination and serological methods. This study generally aimed to molecularly detect and characterize A. marginale in cattle and ticks in Cebu, Philippines. A total of 12 bovine blood samples and 60 Rhipicephalus (Boophilus) microplus ticks were collected on the Cebu Island in 2011. 16S rRNA-based screening-PCR and DNA sequencing revealed 8 cattle (66.7%) and 8 ticks (13.3%) to be positive for A. marginale, and 1 tick (1.7%) to be positive for A. centrale. Selected positive DNA samples were further characterized based on 16S rRNA (longer sequence), Msp5, Msp1α,gltA and groEL genes for phylogenetic analyses. Sequence identities of partial DNA fragments of A. marginale from the Philippines revealed 99.1–100% (16S rRNA, gltA, groEL and Msp5) and 94.3–97.6% (Msp1α) identities to the closest isolates from other countries. Moreover, sequence analysis of the Msp1α, gene showed 3 variants, including a case of co-infection with 2 variants. Phylogenetic analyses based on Msp1α and Msp5 genes revealed that Philippine A. marginale isolates formed a monophyletic lineage, and were phylogenetically related to Brazilian and Chinese isolates. On the other hand, a highly specific and sensitive nested PCR based on groEL, with a detection limit of 2 copies/PCR, was developed to detect A. marginale in the Philippines. This study reported the first molecular detection and characterization of A. marginale in cattle and R. microplus in Cebu, Philippines.

【 授权许可】

Unknown