Molecular identification of Rickettsia spp. in chigger mites in Taiwan.

The genus Rickettsia is the causative agent of several rickettsial diseases that are primarily transmitted by hard ticks. The occurrence of Rickettsia in chigger mites, which are vectors of scrub typhus in the western Pacific region, has been infrequently investigated. We identified Rickettsia spp. in chiggers collected from small mammals in six counties of Taiwan. Moreover, by capitalising on parallel Rickettsia detections on small mammals and their infested ticks and fleas, we were able to identify Rickettsia spp. that suggested more intimate associations with chigger mites. Rickettsia detection rates in 318 pools of chiggers were 21.7% and 22.3% when based on the ompB and gltA gene, respectively. Overall, we identified six (based on the ompB gene) and eight (gltA gene) Rickettsia species. Approximately half of the sequenced species were most similar to Rickettsia sp. clone MB74-1 (ompB gene) and Rickettsia sp. TwKM02 (gltA gene). Furthermore, both species were either infrequently or never identified in small mammals, ticks and fleas, which suggests that chigger mites might be the primary host of both rickettsiae. Whether both species are pathogenic to humans remains to be studied. They may also be microbial endosymbionts of chigger mites, with their potential effects on the pathogenicity of the aetiologic agent of scrub typhus deserving further investigations.

Molecular detection of Rickettsia species and host associations of Laelaps mites (Acari: Laelapidae) in Taiwan.

Various rickettsiae have recently been detected in Laelaps mites (Acari: Laelapidae), which are common ectoparasites of rodents; however, investigations on this topic remain very scarce, particularly in Asia. In the present study, shrews and rodents were trapped from 2006 to 2010 in eight lowland regions of Taiwan (< 500 m in elevation) to collect associated Laelaps mites, from which Rickettsia-a group of emerging pathogens-were detected and identified by assaying the gltA and ompB genes. A total of 853 Laelaps mites of at least four species were collected from a sample of 1004 small mammals that included one shrew and 10 rodent species. Rattus losea was the most common species (44.9% of total hosts) and hosted the highest percentage of mites (76.6% of total mites). Laelaps nuttalli was the most abundant mite species (51.7% of total mites), followed by Laelaps echidninus (24.2%), Laelaps sedlaceki (23.1%), and Laelaps myonyssognathus (0.2%). Notably, Rickettsia species with the highest similarity to spotted fever group (SFG) rickettsiae were identified from seven of the 72 pools of Laelaps mites. The presence of SFG rickettsiae in hematophagous Laelaps mites, particularly including species that are closely associated with commensal rodents in frequent contact with humans, calls for further investigation on the competence of Laelaps mites in transmitting rickettsiae.

DNMT3L promotes quiescence in postnatal spermatogonial progenitor cells.

The ability of adult stem cells to reside in a quiescent state is crucial for preventing premature exhaustion of the stem cell pool. However, the intrinsic epigenetic factors that regulate spermatogonial stem cell quiescence are largely unknown. Here, we investigate in mice how DNA methyltransferase 3-like (DNMT3L), an epigenetic regulator important for interpreting chromatin context and facilitating de novo DNA methylation, sustains the long-term male germ cell pool. We demonstrated that stem cell-enriched THY1(+) spermatogonial stem/progenitor cells (SPCs) constituted a DNMT3L-expressing population in postnatal testes. DNMT3L influenced the stability of promyelocytic leukemia zinc finger (PLZF), potentially by downregulating Cdk2/CDK2 expression, which sequestered CDK2-mediated PLZF degradation. Reduced PLZF in Dnmt3l KO THY1(+) cells released its antagonist, Sal-like protein 4A (SALL4A), which is associated with overactivated ERK and AKT signaling cascades. Furthermore, DNMT3L was required to suppress the cell proliferation-promoting factor SALL4B in THY1(+) SPCs and to prevent premature stem cell exhaustion. Our results indicate that DNMT3L is required to delicately balance the cycling and quiescence of SPCs. These findings reveal a novel role for DNMT3L in modulating postnatal SPC cell fate decisions.

Widespread Rickettsia spp. Infections in Ticks (Acari: Ixodoidea) in Taiwan.

Ticks are second to mosquitoes as the most important disease vectors, and recent decades have witnessed the emergence of many novel tick-borne rickettsial diseases, but systematic surveys of ticks and tick-borne rickettsioses are generally lacking in Asia. We collected and identified ticks from small mammal hosts between 2006 and 2010 in different parts of Taiwan. Rickettsia spp. infections in ticks were identified by targeting ompB and gltA genes with nested polymerase chain reaction. In total, 2,732 ticks were collected from 1,356 small mammals. Rhipicephalus haemaphysaloides Supino (51.8% of total ticks), Haemaphysalis bandicota Hoogstraal & Kohls (28.0%), and Ixodes granulatus Supino (20.0%) were the most common tick species, and Rattus losea Swinhoe (44.7% of total ticks) and Bandicota indica Bechstein (39.9%) were the primary hosts. The average Rickettsia infective rate in 329 assayed ticks was 31.9% and eight Rickettsia spp. or closely related species were identified. This study shows that rickettsiae-infected ticks are widespread in Taiwan, with a high diversity of Rickettsia spp. circulating in the ticks. Because notifiable rickettsial diseases in Taiwan only include mite-borne scrub typhus and flea-borne murine typhus, more studies are warranted for a better understanding of the real extent of human risks to rickettsioses in Taiwan.

Cascading effect of economic globalization on human risks of scrub typhus and tick-borne rickettsial diseases.

The increase in global travel and trade has facilitated the dissemination of disease vectors. Globalization can also indirectly affect vector-borne diseases through the liberalization of cross-border trade, which has far-reaching, worldwide effects on agricultural practices and may in turn influence vectors through the modification of the ecological landscape. While the cascading effect of economic globalization on vector-borne diseases, sometimes acting synergistically with regional agricultural policy, could be substantial and have significant economic, agricultural, and public health implications, research into this remains very limited. We evaluated how abandonment of rice paddies in Taiwan after joining the World Trade Organization, along with periodic plowing, an agricultural policy to reduce farm pests in abandoned fields can unexpectedly influence risks to diseases transmitted by ticks and chiggers (larval trombiculid mites), which we collected from their small-mammal hosts. Sampling was limited to abandoned (fallow) and plowed fields due to the challenge of trapping small mammals in flooded rice paddies. Striped field mice (Apodemus agrarius) are the main hosts for both vectors. They harbored six times more ticks and three times more chiggers in fallow than in plowed plots. The proportion of ticks infected with Rickettsia spp. (etiologic agent of spotted fever) was three times higher in fallow plots, while that of Orientia tsutsugamushi (scrub typhus) in chiggers was similar in both treatments. Fallow plots had more ground cover and higher vegetation than plowed ones. Moreover, ticks and chiggers in both field types were dominated by species known to infest humans. Because ticks and chiggers should exhibit very low survival in flooded rice paddies, we propose that farm abandonment in Taiwan, driven by globalization, may have inadvertently led to increased risks of spotted fever and scrub typhus. However, periodic plowing can unintentionally mitigate vector burdens. Economic globalization can have unexpected consequences on disease risk through modification of the agricultural landscape, but the outcome may also be influenced by agricultural policies, calling for further research on vector-borne diseases and their control from broader perspectives.

Fleas of Shrews and Rodents in Rural Lowland Taiwan.

Fleas transmit a variety of pathogens to humans but are relatively understudied in comparison to mosquitoes and ticks, including in Taiwan, where fleas in rural lowlands have never been systematically surveyed. In total, 700 fleas of four species were collected from 1,260 shrews and rodents at nine counties across lowland Taiwan. Nosopsyllus nicanus Jordan (Siphonaptera: Ceratophyllidae) and Xenopsylla cheopis Rothschild (Siphonaptera: Pulicidae) were the most abundant flea species (79.0 and 14.6% of total fleas, respectively); the former was largely limited to the islets, while the latter was restricted to the Taiwan main island. Rattus losea Swinhoe (Rodentia: Muridae) was the most common small mammal species (49.3% of total) and hosted the majority of fleas (88.3% of total). Five Rickettsia spp., including Rickettsia conorii Brumpt (Rickettsiales: Rickettsiaceae), Rickettsia felis Bouyer et al. Rickettsia japonica Uchida, Rickettsia raoultii Mediannikov, and Rickettsia rickettsii Brumpt or closely related species, were identified from 67 individually assayed fleas based on ompB and gltA genes. Rickettsia felis, mainly transmitted by fleas, was detected in one X. cheopis in southern Taiwan where a confirmed human case of infection with R. felis has been reported. The presence of R. felis, along with the other four tick-borne Rickettsia spp., demonstrates that a variety of rickettsiae circulate in rural lowland Taiwan and could pose risks to human health.

Surveillance of potential hosts and vectors of scrub typhus in Taiwan.

BACKGROUND: Scrub typhus is a lethal infectious disease vectored by larval trombiculid mites (i.e. chiggers) infected with Orientia tsutsugamushi (OT) and recent decades have witnessed an emergence of scrub typhus in several countries. Identification of chigger species and their vertebrate hosts is fundamental for the assessment of human risks to scrub typhus under environmental changes, but intensive and extensive survey of chiggers and their hosts is still lacking in Taiwan. METHODS: Chiggers were collected from shrews and rodents in nine counties of Taiwan and were assayed for OT infections with nested polymerase chain reaction (PCR). PCR products were further sequenced to reveal probable OT strains. Rodents were assessed for OT exposure by immunofluorescent antibody assay. Lastly, incidence rate of scrub typhus in each county was associated with loads and prevalence of chigger infestations, seropositivity rate in rodents, and OT positivity rate in chiggers. RESULTS: Rattus losea was the most abundant (48.7% of 1,285 individuals) and widespread (occurred in nine counties) small mammal species and hosted the majority of chiggers (76.4% of 128,520 chiggers). Leptotrombidium deliense was the most common (64.9% of all identified chiggers) and widespread (occurred in seven counties) chigger species but was replaced by Leptotrombidium pallidum or Leptotrombidium scutellare during the cold seasons in two counties (Matsu and Kinmen) where winter temperatures were lower than other study sites. Seropositivity rate for OT exposure in 876 assayed rodents was 43.0% and OT positivity rate in 347 pools of chiggers was 55.9%, with 15 OT strains identified in the 107 successfully sequenced samples. Incidence rate of scrub typhus was positively correlated with chigger loads, prevalence of chigger infestations, seropositivity rate but not OT positivity rate in chiggers. CONCLUSIONS: Our study reveals R. losea as the primary host for chiggers and there exists a geographical and seasonal variation in chigger species in Taiwan. It also emphasizes the importance of recognition of chigger vectors and their vertebrate hosts for a better prediction of human risks to scrub typhus under rapid environmental changes.

The presence of Borrelia valaisiana-related genospecies in ticks and a rodent in Taiwan.

A field survey was conducted to investigate the presence of Borrelia burgdorferi sensu lato (s.l.) in six counties of Taiwan. Spirochetes were successfully isolated from one rodent ear sample out of 485 rodent ears and 53 live, fed tick (Ixodes granulatus) samples. The spirochetes were confirmed to be B. burgdorferi s.l. by real-time PCR. In addition, 23 of 113 tick samples were tested positive for Borrelia DNA according to real-time PCR. The Borrelia isolate from the rodent and the 23 Borrelia DNA samples from the ticks were identified as B. valaisiana-related genospecies by phylogenetic analysis based on flagellin gene sequences. These findings suggest that the Borrelia valaisiana-related strains are maintained in a zoonotic cycle between tick vectors and reservoir hosts in Taiwan.