Efficiency of transovarial transmission of Rickettsia tsutsugamushi in Leptotrombidium arenicola (Acari: Trombiculidae).
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Investigations of trombiculid mites and Rickettsia tsutsugamushi in wild rodents were made in southern Gifu Prefecture where patients infected with tsutsugamushi disease recently have been found. A total of 16,396 trombiculid mites, consisting of 10 species from three genera, was collected from 170 Apodemus speciosus in two locations. Kani-Sakahogi and Kuze. Leptotrombidium scutellare (Nagayo et al.) (44.0%) was most predominant, followed by L. pallidum (Nagayo et al.) (26.9%); L. fuji (Kuwata et al.) (13.6%); and Gahrliepia saduski Womersley (14.2%). These four species constituted the bulk of the chigger mite fauna. L. scutellare was present from October to February with a remarkably high peak in November, whereas pallidum occurred from November to March with the highest peak in December. L. fuji and G. saduski showed their highest peaks in December and moderate peaks in early summer (April and May). Positive identification of Rickettsia tsutsugamushi in wild rodents from Kani-Sakahogi were found to be 50 and 58.3% in November 1985 and 1986, respectively, and 38.5% in November 1986 from Kuze. R. tsutsugamushi was isolated from chigger mites of an L. pallidum-rich group, displaying the highest titer to Karp strain. Serological investigation of rodents to R. tsutsugamushi antibodies were calculated as 41.6 and 50% positive in November 1985 and 1986 in Kani-Sakahogi, respectively, and 50% in November 1986 in Kuze. The Karp strain was dominant in specificity to antibodies. These results indicate that the surveyed areas have a high probability of occurrence of tsutsugamushi disease, and L. scutellare and L. pallidum may serve as the vectors in these areas. Particularly, we suggest that L. scutellare is the most important vector which has caused a recent outbreak of this disease in southern Gifu Prefecture.
In the Philippine Islands, the genus Leptotrombidium currently consists of members of the subgenera Leptotrombidium and Trombiculindus. The following species are described as new: L. (L.) macacaphilus, L. (L.) longimedian, L. (L.) mindanensis, L. (L.) urogale, L. (L.) oculascutum, L. (T.) roseannleilaniae. The distribution of, and comments on, the known vectors of Rickettsia tsutsugamushi, L. (L.) deliense and L. (L.) fletcheri and the previously reported L. (L.) sandfordi, are given. Information on species of the potential scrub itch genera Blankaartia, Eutrombicula, and Schoengastia and a key to the genera and species of medically important trombiculids is provided.
Larvae of Leptotrombidium pallidum (Nagayo, Miyagawa, Mitamura & Tamiya) from uninfected laboratory colonies were fed on mice infected with Rickettsia tsutsugamushi (Hayashi) Ogata. Infection of the chiggers with R. tsutsugamushi was determined by passage of chigger exudates into ddY mice. The passage method was modified so that an inoculum was considered to be positive when R. tsutsugamushi or anti-R. tsutsugamushi antibody, or both, were detected in mice up to the third blind passage. R. tsutsugamushi was detected in six of 18 larvae (33.3%) and in all developmental stages. In adults, five of 18 males and 10 of 46 females were infected with R. tsutsugamushi. In L. fuji (Kuwata, Berge & Philip), R. tsutsugamushi was not found in 57 engorged larvae fed on rickettsemic mice but was found in a very low percentage of deutonymphs and adults. Female L. pallidum from larvae fed on infected mice were paired individually, and F1 larvae were collected. Although eight females were found to be positive for R. tsutsugamushi, the rickettsia was not detected in 12 pools (249 larvae) of F1 larvae from these infected females. We concluded that uninfected mites became infected by feeding on rickettsemic mice at comparatively high rates depending on the species and transmitted this infection transstadially to succeeding life stages, but not vertically to larvae in the following F1 generation.
The ecology of Orientia tsutsugamushi (Hayashi) was studied in chiggers and small animals in an orchard near Bangkok, Thailand. Small animals were trapped monthly between July and November 1992 and examined for the presence of O. tsutsugamushi and ectoparasitic chiggers. A total of 40 Rattus rattus (L.) and 16 Tupaia glis (Diard) was trapped. O. tsutsugamushi was isolated from liver and spleen samples of 30.8% of R. rattus and 18.6% of T. glis. Antibodies to O. tsutsugamushi were detected in 95% of R rattus, and IgG antibodies persisted for up to 10 mo after removing rats from potential reinfection in the field. A total of 1,133 chiggers was identified and examined for the presence of O. tsutsugamushi using a direct fluorescent antibody test, and 2.6% Leptotrombidium deliense Walch, 5.1% Aschoshoengastia indica Hirst, 2.6% A. (Laurentella) sp. #2 and 0.9% A. (Laurentella) sp. #4 were infected. Forty-four pools of chiggers from these animals were triturated and injected into mice. Seven pools were obtained from T. glis and 1 was positive for O. tsutsugamushi, and 3 of 37 pools from R. rattus were positive. A proportion of the engorged chiggers collected was reared to the adult stage, and the progeny of these adults tested for the presence of O. tsutsugamushi. The progeny of 186 females was tested, and the progeny of 2 L. deliense was found to be naturally infected with O. tsutsugamushi.
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Recent studies of Rickettsia tsutsugamushi have demonstrated clearly the phenotypic and genotypic differences between this microorganism and other species belonging to the genus Rickettsia. Therefore, classification of R. tsutsugamushi in a new genus, Orientia gen. nov., is proposed.
Two cases of tsutsugamushi disease misdiagnosed initially as drug eruption were reported. Clinical symptoms of both cases disappeared dramatically after starting minocycline. Statistical examinations were performed on 29 cases of tsutsugamushi disease, including those observed in Mie Prefecture since 1982. Half of them were seen in the last 2 years. The onset was predominantly recorded in November (59%). Presumptive sites of infection were forests (63%) and fields (21%). No patients were infected along river-banks.
In Tottori Prefecture, we gathered the data of all patients known to have been infected with tsutsugamushi disease (TD) over a span of 49 years, from 1950 to 1998. The total number of patients reported so far has been 19, 16 of which occurred after 1988. Recently, it has been observed that the incidence tended to increase. Of the 13 laboratory confirmed patients, 7 came from Saji Village and Chizu Town, in the Yazu district, a place whereby, a body of water known as the Sendai River partly passes through, and is located in the eastern region of Tottori Prefecture. The remaining 6 came from Nichinan Town in the Hino district, a locality whereby the Hino River partly passes through; in the western part of the same prefecture. Both are mountainous areas, where corniferous trees such as Japanese cedars and pines grow. These results indicate that the occurrence or prevalence of TD in Tottori Prefecture was restricted only to particular places. Seasonal occurrence was during April and May in Nichinan, while October and November in Yazu, particularly in Saji, with one exceptional case in Chizu in which TD occurred during spring. It is likely that the causative chigger mite is the Leptotrombidium pallidum (L. pallidum). The causative agent in Yazu was the Gilliam strain only of Orientia tsutsugamushi (O. tsutsugamushi), while in the latter, it was the Karp, Kato or some unknown strains with antigen(s) common to the Gilliam, Karp and Kato strains. The seasonal difference in the occurrence among the three places may be caused by the difference in temperature because the average autumn/spring temperature difference in the 2 districts was 2-3 degrees C lower in Nichinan than in the Saji-Chizu areas. As a result, the activity of larval chigger mites may be temporarily halted or stopped by the relatively lower temperature in Nichinan during autumn.
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Rickettsia tsutsugamushi strains from three recent patients of Tsutsugamushi disease in Niigata Prefecture were isolated primarily in mice and then in L cell cultures. By this procedure, low virulent strains to mice, as well as high virulent ones, could be isolated and cultivated serially in L cell cultures, suggesting the usefulness of L cells for isolation of this species of rickettsia. Each newly isolated strain was identified as a member of R. tsutsugamushi from the results of cross immunological tests and morphological observation. On the other hand, it was recognized that one of these rickettsiae showed immunological properties distinguishable from the prototype strains of Kato, Karp, and Gilliam by the cross complement fixation test, and also had low virulence in mice.
In order to clarify the epidemiological background of the endemic occurrence of tsutsugamushi disease in Toyama Prefecture, Japan, since 1978, comparative surveys have been carried out between endemic and nonendemic areas. Rickettsia tsutsugamushi (Rt) was isolated at a rate of about 36% (158/439) from field rodents in the endemic area while it was not isolated from any of 280 in nonendemic areas. In all of six stations in the endemic area, a significantly high proportion of rodents were found to be Rt carriers. However, no Rt was isolated from rodents captured from July to September. The organism was isolated from rodents captured in the other months, especially in a high proportion in November when infestation of rodents with Leptotrombidium pallidum was at its peak. When the rodents were examined by indirect immunofluorescence staining, the rate of anti-Rt antibody-positive animals was about 55% (157/287) and about 17% (62/368) in endemic and nonendemic areas, respectively. Larvae of mites collected from the rodents were found to belong to four genera and 11 species. Among them L. pallidum was the only mite that had been known to be a vector of Rt. L. pallidum was found most frequently and in abundance from rodents in the endemic area, whereas it was present in very small numbers in rodents in nonendemic areas. The infestation of rodents with L. pallidum showed a seasonal variation, i.e. two peaks per year, in spring and autumn, and the number of mites detected was markedly greater in November than in spring. Rt was isolated from L. pallidum on rodents captured in the endemic area.
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With a view to clarifying the actual state of inapparent infection of tsutsugamushi diseases, inhibitants of endemic and nonendemic areas were screened for anti-Rickettsia tsutsugamushi antibody (anti-Rt antibody) by the indirect immunofluorescence test. The anti-Rt antibody-positive rate in the inhabitants of the endemic area (about 50%) was statistically significantly higher than that in the nonendemic area (14.7%). The antibody titer in the inhabitants of the endemic area was 10-160, and the number of inhibitants showing a high antibody titer was 2-4 times larger than that of the nonendemic area. A total of 257 volunteers in the endemic area were analyzed for the changes in anti-Rt antibody titer over 1.5-2 years on an individual basis. An increase in the antibody titer was found in 20 inhabitants. There was no difference in the anti-Rt antibody-positive rate between male and female in either the endemic or the nonendemic area. The positive rate was also compared as to the distribution by 10 years of age. In the endemic area, there were no significant differences in the positive rate between any pair of 10-year age groups from 30s to 60s, whereas in the nonendemic area, the positive rate in the teen-age group was significantly lower than those in the age groups of 20 years or older. In Yamada district, the numbers of serum samples obtained from each age group were about the same, and the distribution of the positive rates showed a normal distribution. The nurse students having their homes in Toyama Prefecture were plotted on the map as for their anti-Rt antibody and geographical distribution. The results showed that many of them having homes in the endemic area were positive for the antibody, while some antibody-positives were scattered all over Toyama Prefecture.
Embryos of Leptotrombidium (Leptotrombidium) pallidum mites naturally infected with Rickettsia tsutsugamushi were examined by electron microscopy. Rickettsiae were not found in eggs just after oviposition, but were easily detected in cells at the various parts of the embryos just before hatching, indicating that the rickettsiae are surely vertically transmitted from infected adult mites to the larvae through embryos, and the rickettsiae may multiply in situ during the developing process of the embryo.