Novel Genetic Variants of Anaplasma phagocytophilum , Anaplasma bovis , Anaplasma centrale , and a Novel Ehrlichia sp. in Wild Deer and Ticks on Two Major Islands in Japan

Abstract

Wild deer are one of the important natural reservoir hosts of several species of Ehrlichia and Anaplasma that cause human ehrlichiosis or anaplasmosis in the United States and Europe. The primary aim of the present study was to determine whether and what species of Ehrlichia and Anaplasma naturally infect deer in Japan. Blood samples obtained from wild deer on two major Japanese islands, Hokkaido and Honshu, were tested for the presence of Ehrlichia and Anaplasma by PCR assays and sequencing of the 16S rRNA genes, major outer membrane protein p44 genes, and groESL. DNA representing four species and two genera of Ehrlichia and Anaplasma was identified in 33 of 126 wild deer (26%). DNA sequence analysis revealed novel strains of Anaplasma phagocytophilum, a novel Ehrlichia sp., Anaplasma centrale, and Anaplasma bovis in the blood samples from deer. None of these have been found previously in deer. The new Ehrlichia sp., A. bovis, and A. centrale were also detected in Hemaphysalis longicornis ticks from Honshu Island. These results suggest that enzootic cycles of Ehrlichia and Anaplasma species distinct from those found in the United States or Europe have been established in wild deer and ticks in Japan.

Detection of Anaplasma phagocytophilum and Anaplasma bovis in Small Wild Mammals from Taichung and Kinmen Island, Taiwan

“…In 1990, A. phagocytophilum was identified as a possible cause of human granulocytic anaplasmosis (HGA) in the United States (3). In Asia, A. phagocytophilum-infected ticks, domestic animals, and wild animals have been detected in China, Russia, Korea, and Japan (4)(5)(6)(7)(8)(9)(10)(11)(12). However, till date, there is no epidemiological evidence of A. phagocytophilum infection in Taiwan.…”

“…However, these sequences branched out from those of A. phagocytophilum that were detected in a patient from the United States (AF093789), a horse from Sweden (AY527214), Ixodes ovatus from Japan (AY969012), and I. persulcatus from Russia (HM366590), as well as related sequences detected in Dermacentor silvarum (DQ449948), A. agrarius (DQ342324, GQ412337), Tscherskia triton (GQ412339), and a sheep (GQ412338) from Jilin province in northeastern China (5,14). A. phagocytophilum-related sequences detected in wild rodents from Taiwan were distinct from those detected in cattle from Yonaguni Island (EU368727, EU368728), deer (AB196720) and boars in Japan, and goats (JN558811) in China (6,13,21,22).…”

“…The remaining 25 sequences detected in R. losea (AB817955, AB817956, AB817957, and AB817959), A. agrarius (AB817958), B. indica, and M. caroli were identified as those of A. bovis and branched out from a sequence isolated from a dog from South Africa (U03775), a goat from China (JN558819, JN558822, JN558828) (21), deer and boars from Japan (AB196475, AB588967, AB588965, AB588966, AB588970, AB588971), and cattle (EU368731, EU368730) from Yonaguni Island, the southernmost island of Japan (6,7,13,22,23). Liu et al (21) reported that A. bovis was detected in monocytes isolated from sheep experimentally inoculated with blood from a goat from Suizhou, Hubei province, China.…”

“…DNA was extracted from the spleen using the QuickGene-800 nucleic acid isolation system (Kurabo, Osaka, Japan) (13). Anaplasma-specific DNA was detected using nested PCR targeting 16S rDNA with the forward primer EC9 and the reverse primer EC12A in the first step of the PCR reaction (6). The initial amplification consisted of 40 cycles, each consisting of 60 s at 949 C, 60 s at 589 C, and 90 s at 729 C. For nested PCR, the forward primer Abpp was annealed to DNA sequences of A. bovis, A. phagocytophilum, and A. platys, and a second forward primer, Acom, was annealed to the sequences of A. centrale, A. ovis, and A. marginale.…”

Molecular Evidence forAnaplasma bovisInfection in Wild Reeves' Muntjac (Muntiacus reevesi), Southwest China

“…Clinical signs of A. bovis infection consist of sporadic fever lasting for several weeks, anemia, weight loss, and possibly death of naive or stressed animals (Sreekumar et al 1996;Liu et al 2012). In recent years, molecular evidence of A. bovis infection has been described in domestic and wild animals, especially in cervidae (Kawahara et al 2006;Masuzawa et al 2011). There have been no reports of cervids infected with this pathogen in China, despite a high prevalence of A. bovis (16%) in goats from central and southern China (Liu et al 2012).…”