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

Abstract

ABSTRACT. 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.

Molecular epidemiological survey and genetic analysis of vector-borne infections of cattle in Luzon Island, the Philippines

“…A. marginale and B. bovis were screened using a nested PCR assay based on 16S rRNA (Ybañez et al, 2012) and rhoptry-associated protein (RAP)-1 (Figueroa et al, 1993), respectively. B. bigemina, Theileria spp., and T. evansi were detected using single-step PCR methods based on apical membrane antigen (AMA)-1 (Sivakumar et al, 2012), major piroplasm surface protein (MPSP) (Kakudai et al, 1998) and RoTat1.2 (Claes et al, 2004) genes, respectively. All PCR assays were conducted as previously described with slight modifications.…”

Molecular detection and characterization of Theileria species in the Philippines

“…The two diseases of major concern to Philippine authorities are babesiosis caused by Babesia bigemina, B. bovis and B. major (Bock et al 2004) and anaplasmosis caused by Anaplasma marginale and A. centrale (Ybañez et al 2013), collectively producing tick fever transmitted by Rhipicephalus (Boophilus) microplus. Rhipicephalus (Boophilus) microplus is known to transmit Theileria parasites in South Asian countries but in the Philippines there no study that this tick species transmit Theileria parasites.…”

High Genetic Diversity of <i>Anaplasma marginale</i> Detected from Philippine Cattle

“…Previously, only few cattle in a limited area were used to demonstrate the genotypes of A. marginale in the Philippines [32]. Thus, the present study was endeavored to determine the genetic diversity of A. marginale using more number of bovine blood samples collected from different geographic locations in the Philippines.…”