Hiromi Fujita

Prevalence and Genetic Diversity of Bartonella Species Isolated from Wild Rodents in Japan

ABSTRACT Here, we describe for the first time the prevalence and genetic properties of Bartonella organisms in wild rodents in Japan. We captured 685 wild rodents throughout Japan (in 12 prefectures) and successfully isolated Bartonella organisms from 176 of the 685 rodents (isolation rate, 25.7%). Those Bartonella isolates were all obtained from the rodents captured in suburban areas (rate, 51.8%), but no organism was isolated from the animals captured in city areas. Sequence analysis of rpoB and gltA revealed that the Bartonella isolates obtained were classified into eight genetic groups, comprising isolates closely related to B. grahamii (A-I group), B. tribocorum and B. elizabethae (B-J group), B. tribocorum and B. rattimassiliensis (C-K group), B. rattimassiliensis (D-L group), B. phoceensis (F-N group), B. taylorii (G-O group), and probably two additional novel Bartonella species groups (E-M and H-P). B. grahamii, which is one of the potential causative agents of human neuroretinitis, was found to be predominant in Japanese rodents. In terms of the relationships between these Bartonella genetic groups and their rodent species, (i) the A-I, E-M, and H-P groups appear to be associated with Apodemus speciosus and Apodemus argenteus; (ii) the C-K, D-L, and F-N groups are likely implicated in Rattus rattus; (iii) the B-J group seems to be involved in Apodemus mice and R. rattus; and (iv) the G-O group is probably associated with A. speciosus and Clethrionomys voles. Furthermore, dual infections with two different genetic groups of bartonellae were found in A. speciosus and R. rattus. These findings suggest that the rodent in Japan might serve as a reservoir of zoonotic Bartonella infection.

Isolation and characterization of a novel Borrelia group of tick-borne borreliae from imported reptiles and their associated ticks.

The members of the genus Borrelia are transmitted by arthropods and known to be infectious to vertebrates. Here we found isolates and DNAs belonging to the Borrelia turcica and unknown Borrelia species from imported reptiles and their ectoparasites. The Borrelia strains were isolated from blood and multiple organs of exotic tortoises, and were experimentally infectious to captive-bred tortoises. These findings suggest that these tortoises may be a candidate as the reservoir host of the Borrelia species. In this study, the Borrelia strains were also isolated from and/or detected in hard-bodied ticks, Amblyomma ticks and Hyalomma ticks. In some of these ticks, immunofluorescence imaging analysis revealed that the Borrelia had also invaded into the tick salivary glands. Accordingly, these ticks were expected to be a potential vector of the Borrelia species. Sequencing analyses of both housekeeping genes (flaB gene, gyrB gene and 16S rDNA gene) and 23S rRNA gene-16S rRNA gene intergenic spacer region revealed that these Borrelia strains formed a monophyletic group that was independent from two other Borrelia groups, Lyme disease Borrelia and relapsing fever Borrelia. From these results, the novel group of Borrelia comprises the third major group of arthropod-transmitted borreliae identified to date.

Molecular Identification of Ehrlichia Species and 'Candidatus Neoehrlichia mikurensis' from Ticks and Wild Rodents in Shizuoka and Nagano Prefectures, Japan

A total of 293 ticks and 111 wild rodents that were collected in Shizuoka and Nagano Prefectures, Japan, were examined for infection of Ehrlichia species and ‘Candidatus Neoehrlichia mikurensis.’ The 16S rDNA or the omp‐1 gene of these bacterial DNAs were detected from the spleens of tick‐inoculated mice (5 positive/total 29 mice) or from the spleens of wild rodents (25 positive/total 111 rodents) by PCR amplification. Sequencing of the 16S rDNA revealed Ehrlichia spp. from the 5 tick‐inoculated mice and 8 wild rodents, and ‘Candidatus N. mikurensis’ from 17 wild rodents. The data suggest the presence of additional genetic variants, and potential vectors and/or reservoirs for these bacteria in central Japan.

Multilocus Sequence Typing Implicates Rodents as the Main Reservoir Host of Human-Pathogenic Borrelia garinii in Japan

ABSTRACT Multilocus sequence typing of Borrelia garinii isolates from humans and comparison with rodent and tick isolates were performed. Fifty-nine isolates were divided into two phylogenetic groups, and an association was detected between clinical and rodent isolates, suggesting that, in Japan, human-pathogenic B. garinii comes from rodents via ticks.

Presence of a novel ehrlichia sp. in ixodes granulatus found in okinawa, japan

Ehrlichia‐specific DNA fragments of Ehrlichia omp‐1 and groEL genes were found in two I. granulatus ticks which had been collected from wild small mammals in a subtropical zone in Japan. The DNA sequences of groEL and 16SrDNA of the suspected Ehrlichia were clustered into a group of E. chaffeensis, E. muris, and Ehrlichia sp. HF565 found in I. ovatus, but were distinctly different. Therefore the Ehrlichia strain was designated as a novel Ehrlichia sp. 360. The Ehrlichia sp. 360 was detected in I. granulatus but not in any other ticks. This suggests that I. granulatus is a probable vector of Ehrlichia sp. 360 in Japan.

Characterization of reptile-associated Borrelia sp. in the vector tick, Amblyomma geoemydae, and its association with Lyme disease and relapsing fever Borrelia spp.

The genus Borrelia is arthropod-borne infectious agents in vertebrates, and is classified into Lyme disease (LD) Borrelia spp. and Relapsing fever (RF) Borrelia spp. In addition to these Borrelia groups, we recently reported reptile-associated (REP) Borrelia spp. from reptiles and from hard-bodied ticks, which probably transmitted the REP Borrelia spp. In this study, we investigated the presence of REP Borrelia sp. in moulted ticks, and found that trans-stadial transmission of REP Borrelia sp. occurred in the midgut, while it was observed that REP Borrelia sp. entered the salivary gland during blood-feeding. This characteristic is also found in LD Borrelia spp., which are also transmitted by hard-bodied ticks. Although phylogenetic analysis demonstrated that REP Borrelia spp. are similar to RF Borrelia spp., the ecology of the spirochaetes within the vector ticks is different for REP Borrelia spp. and RF Borrelia spp. Elucidation of the evolutionary history of the genus Borrelia and its adaptation to ticks promises to be of great interest to researchers of vector-borne microorganisms.

Diagnostic Assay for Rickettsia japonica

We developed a specific and rapid detection system for Rickettsia japonica and R. heilongjiangensis, the causative agents of spotted fever, using a TaqMan minor groove binder probe for a particular open reading frame (ORF) identified by the R. japonica genome project. The target ORF was present only in R. japonica–related strains.

Detection of Kobe-type Babesia microti associated with Japanese human babesiosis in field rodents in central Taiwan and southeastern mainland China

Field rodent surveys for Babesia infection were performed from 2002 to 2005 in the vicinities of human babesiosis occurrences in Taiwan and mainland China. Babesia microti was identified by microscopical examination and/or PCR in 1 Rattus coxinga and 1 Crocidura horsfieldii in central Taiwan and in 13 Niviventer confucianus and 1 Apodemus agrarius in Zhejiang and Fujian Provinces of southeastern China. Of 15 B. microti samples detected by PCR, all except 1 were shown to be the Kobe-type, the aetiological small subunit rRNA gene-type of the first Japanese patient; the exception was also a Kobe-related type. The Kobe-type had been found in rodents only in a few places including the human infection occurrence place in Japan. The internal transcribed spacer 1 to 2 sequences of the Taiwanese and Chinese Kobe-types were very similar to each other but considerably different (approx. 94% pairwise identities) from that of the Japanese Kobe-type. A Taiwanese Kobe-type strain was serologically differentiated from the Kobe strain originating from the Japanese first patient. The distribution of the Kobe-type in the vicinities of human babesiosis occurrences in Taiwan and China as well as in Japan is suggestive of involvement of the Kobe-type in Asian human babesiosis.

Spotted fever group Rickettsia sp. closely related to Rickettsia japonica, Thailand.

Suggested citation for this article: Takada N, Fujita H, Kawabata H, Ando S, Sakata A, Takano A, et al. Spotted fever group Rickettsia sp. closely related to R. japonica, Thailand [letter]. Emerg Infect Dis [serial on the Internet]. 2009 Apr [date cited]. Available from http://www.cdc.gov/EID/content/15/4/610.htm

Survey of Babesia microti Infection in Field Rodents in Japan : Records of the Kobe-Type in New Foci and Findings of a New Type Related to the Otsu-Type

Of 247 rodents comprising 5 genera and 7 species collected at 17 sites throughout Japan from 2003 to 2005, Babesia microti was detected microscopically and by polymerase chain reaction (PCR) in 36 rodents comprising 2 genera and 3 species from 12 sites. Based on the analysis of small subunit ribosomal RNA gene (SSUrDNA) sequences, the Kobe‐type, the etiological type of the first Japanese case of human infection was found in Apodemus speciosus and Apodemus argenteus in Aomori, the northernmost prefecture of the Japanese mainland, while the U.S.‐type was found on Hokkaido Island and the Otsu‐type was widely distributed. In addition, a new Otsu‐related type was detected exclusively in Eothenomys andersoni in Nagano, a prefecture in central Japan. The sequences of internal transcribed spacer 1 to 2 (ITS1/2) of the present Kobe‐ and Otsu‐types were almost identical to those of the same types previously identified. The ITS1/2 sequence of the U.S.‐type identified in Hokkaido in this survey was somewhat different from that of the U.S.‐type strain originating from the U.S.A., with approximately 95% identity. This value was similar to the 94% identity found between the ITS1/2 sequences of the Otsu‐type and the new Otsu‐related type. The new Otsu‐related type of B. microti was isolated as the Nagano strain, which was serologically differentiated from the other type strains of B. microti. The divergence and distribution of genotypes are important factors in investigating the epidemiology of human B. microti infection in Japan.