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Abstract: An Anaplasma species closely related to Anaplasma phagocytophilum detected in sika deer in Hokkaido, Japan was molecularly analyzed using 16S rRNA, citrate synthase (gltA), and heat-shock operon (groEL) gene sequences. Genome walking was performed to determine its complete gltA and groEL sequences (1233bp and 1650bp, respectively). Percent identities to the closest A. phagocytophilum sequences from the US and European strains were 98.6–98.8%, 76.5%, and 80.3–80.8% for 16S rRNA, gltA, and groEL genes, respectively. For deduced amino acid sequences, percent identities to the closest A. phagocytophilum sequences were 66.7% and 97.6% for gltA and groEL genes, respectively. Phylogenetic analyses revealed divergence from any known A. phagocytophilum strain. The lower identities and the divergent phylogenetic position of the Anaplasma sp. detected from sika deer in Japan with established A. phagocytophilum strains provide evidence of its potential novelty. [Copyright &y& Elsevier]

Theileria orientalis and hemotropic Mycoplasma spp. (hemoplasmas) can cause anemia in cattle. Cattle infected with one of these two pathogens tend to resist infection by the other pathogen. This is called the "interference phenomenon". However, the detailed investigation of this phenomenon using molecular techniques has not been performed until now. We used PCR to analyze blood samples from cattle grazing in two pastures, and investigated interference between T. orientalis and hemoplasmas. Results indicate that single infection with T. orientalis or hemoplasmas was more common than co-infections. The degree of anemia observed in co-infected animals was significantly milder compared to those only infected with T. orientalis. These findings revealed that the interaction between these two pathogens in cattle demonstrated interference., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Recent molecular analyses of the Anaplasma sp. closely related to Anaplasma phagocytophilum (previously believed to be A. phagocytophilum) in Japan have clarified its distinct phylogenetic position. PCR methods relying on 16S rRNA- and P44/MSP2-based primers designed to detect this species have low sensitivity and specificity. In this study, a highly sensitive and specific nested PCR method using newly designed primers based on heat-shock operon gene (groEL) was developed to detect this species. The method was later used in an epidemiological study testing DNA samples from 85 Ixodid ticks (collected by flagging) and 50 cattle from the same pastureland in Nakaosobetsu, Hokkaido, Japan. Results revealed prevalence rates of 2.4% (2 of 85) in ticks and 2% (1 of 50) in cattle. The present study also reported the first molecular detection of the Anaplasma sp. closely related to A. phagocytophilum in Japan in H. douglasii, and established a new reliable PCR method that detects this Anaplasma sp. closely related to A. phagocytophilum in Japan.

We describe here the clinical significance of coinfection with Theileria orientalis and Babesia ovata in cattle. Anemia status in a herd of dairy cattle in Japan was investigated in relation to infection with these parasites. Our findings indicate that while B. ovata infection might not be the primary cause of anemia in the cattle, it may contribute to the clinical development of anemia in animals coinfected with both B. ovata and T. orientalis.