The chigger mite genus Leptotrombidium Nagayo, Miyagawa, Mitamura and Imamura, 1916 is reviewed using literature data. For 340 larval species brief diagnoses, synonymy, data on type hosts and type localities are provided. The genus is divided into species-groups based on morphological evidence enabling easier establishment of group-membership of unknown specimens in the future. Some species groups are supported by a hierarchical cluster analysis with multiscale bootstrap resampling applied to a matrix including 335 species and geographic morphotypes and 19 standard quantitative characters. Six new species from mammalian hosts are described: L. aenigmami sp. nov., L. abramovi sp. nov., L. tikhonovi sp. nov., L. bochkovi sp. nov., L. laoense sp. nov., and L. megaloti sp. nov. from Laos. Seven names created by Vercammen-Grandjean and Langston (1976) for infrasubspecific entities are applied to species with the same descriptions: Leptotrombidium tenompaki sp. nov., L. kinabalui sp. nov., L. megabodense sp. nov., L. minului sp. nov., L. ului sp. nov., L. megalangati sp. nov., and L. saigoni sp. nov. A new replacement name is proposed: L. ushi nom. nov. pro L. hsui Wu, Yang and Li, 1999 (praeocc. Yu, Yang and Gong, 1986). Nineteen new synonyms and 7 new combinations are proposed: Leptotrombidium (= Hsuella Wang, Li and Shi, 1989, syn. nov.; = Leptotrombidium (Monosigmum) Wen, 2001, syn. nov.), L. deliense (Walch, 1922) (= L. deliense sinense Wen and Chen, 1984, syn. nov.; = L. deliense microsetosa Zhao, Tang and Mo, 1986, syn. nov.), L. sialkotense Vercammen-Grandjean and Langston, 1976 (= L. jishoum Wen, Li, Zhang and Liao, 1988, syn. nov.), L. imphalum Vercammen-Grandjean and Langston, 1976 (= L. imphalum sabahense Vercammen-Grandjean and Langston, 1976, syn. nov.; = L. chiangraiensis Tanskul and Linthicum, 1997, syn. nov.), L. wenense Wu, Wen, Yang and Wu, 1982 (= L. kaohuense Li, Wang and Chen, 1997, syn. nov.), L. longimedian Brown, 1992 (= L. mindanensis Brown, 1992, syn. nov.), L. silvaticum Hushcha and Schluger, 1967 (= L. pakistanum Vercammen-Grandjean and Langston, 1976, syn. nov.), L. cricethrionis Wen, Sun and Sun, 1984 (= L. rusticum Yu, Yang and Gong, 1986, syn. nov.), L. intermedium (Nagayo, Mitamura and Tamiya, 1920) (= Trombicula (L.) daisen Kumada and Sasa, 1953, syn. nov.; = Trombicula hiranumai Kanda, 1942, syn. nov.), L. fletcheri (Womersley and Heaslip, 1943) (= L. fletcheri francolini Wen and Xiang, 1984b, syn. nov.), L. apertum Kudryashova, 1979 (= L. sorosi Kharadov, 1995, syn. nov.; = L. tolaicus Kharadov, 2000, syn. nov.), L. turdicola Vercammen-Grandjean and Langston, 1976 (= L. muntiaci Xiang and Wen, 1984d, syn. nov.; = L. suense Wen, 1984g, syn. nov.), L. paradux Vercammen-Grandjean and Langston, 1976 (= L. montanum Stekolnikov, 2004, syn. nov.), L. hubeiense (Wang, Li and Shi, 1989) comb. nov. from Hsuella, L. dunqingi (Liu, Xiang and Ma, 2003) comb. nov. from Hsuella, L. nainae (Kharadov, 1990) comb. nov. from Montivagum, L. mongolicum (Kudryashova, 1988) comb. nov. from Montivagum, L. kunitzkyi (Kudryashova, 1988) comb. nov. from Montivagum, L. alaicum (Kharadov, 1994) comb. nov. from Montivagum, and Lorillatum nudisensillum (Yu, Gong and Tao, 1981) comb. nov. from Leptotrombidium. A key to Leptotrombidium species is provided.

Conventional gold standard characterization of chigger mites involves chemical preparation procedures (i.e. specimen clearing) for visualization of morphological features, which however contributes to destruction of the arthropod host DNA and any endosymbiont or pathogen DNA harbored within the specimen. In this study, a novel work flow based on autofluorescence microscopy was developed to enable identification of trombiculid mites to the species level on the basis of morphological traits without any special preparation, while preserving the mite DNA for subsequent genotyping. A panel of 16 specifically selected fluorescence microscopy images of mite features from available identification keys served for complete chigger morphological identification to the species level, and was paired with corresponding genotype data. We evaluated and validated this method for paired chigger morphological and genotypic ID using the mitochondrial cytochrome c oxidase subunit I gene (coi) in 113 chigger specimens representing 12 species and 7 genera (Leptotrombidium, Ascoschoengastia, Gahrliepia, Walchia, Blankaartia, Schoengastia and Schoutedenichia) from the Lao People’s Democratic Republic (Lao PDR) to the species level (complete characterization), and 153 chiggers from 5 genera (Leptotrombidium, Ascoschoengastia, Helenicula, Schoengastiella and Walchia) from Thailand, Cambodia and Lao PDR to the genus level. A phylogenetic tree constructed from 77 coi gene sequences (approximately 640 bp length, n = 52 new coi sequences and n = 25 downloaded from GenBank), demonstrated clear grouping of assigned morphotypes at the genus levels, although evidence of both genetic polymorphism and morphological plasticity was found. With this new methodology, we provided the largest collection of characterized coi gene sequences for trombiculid mites to date, and almost doubled the number of available characterized coi gene sequences with a single study. The ability to provide paired phenotypic-genotypic data is of central importance for future characterization of mites and dissecting the molecular epidemiology of mites transmitting diseases like scrub typhus.

Background: Scrub typhus, caused by a bacterial pathogen (Orientia spp.), is a potentially life-threatening febrile illness widely distributed in the Asia-Pacific region and is emerging elsewhere. The infection is transmitted by the larval stage of trombiculid mites (“chiggers”) that often exhibit low host specificity. Here, we present an analysis of chigger ecology for 38 species sampled from 11 provinces of Thailand and microbiomes for eight widespread species. Results: In total, > 16,000 individual chiggers were collected from 1574 small mammal specimens belonging to 18 species across four horizontally-stratified habitat types. Chigger species richness was positively associated with higher latitudes, dry seasonal conditions, and host maturity; but negatively associated with increased human land use. Human scrub typhus incidence was found to be positively correlated with chigger species richness. The bacterial microbiome of chiggers was highly diverse, with Sphingobium, Mycobacterium, Neisseriaceae and various Bacillales representing the most abundant taxa. Only Leptotrombidium deliense was found to be infected with Orientia and another potential pathogen, Borrelia spp., was frequently detected in pools of this species. β-diversity, but not α-diversity, was significantly different between chigger species and geographic regions, although not between habitat types. Conclusion: Our study identified several key environmental and host-derived correlates of chigger species richness across Thailand, which in turn impacted on human scrub typhus incidence. Moreover, this first extensive field survey of the chigger microbiome revealed species- and province-level variation in microbial β-diversity across the country, providing a framework for future studies on interactions between pathogens and other symbionts in these understudied vectors.

For over a decade, the presence of trombiculid mites in some mountain areas of La Rioja (Northern Spain) and their association with seasonal human dermatitis have been recognized. This work aimed to establish the species identity of the agent causing trombiculiasis in the study area.

Chigger mites are vectors of the bacterial disease scrub typhus, caused by Orientia spp. The bacterium is vertically transmitted in the vector and horizontally transmitted to terrestrial vertebrates (primarily wild small mammals), with humans as incidental hosts. Previous studies have shown that the size of the chigger populations is correlated with the density of small mammals in scrub typhus-endemic regions. Here, we explore interactions between the small mammals and chiggers in two oil palm plantations located in the Perak and Johor states of Peninsular Malaysia. The location in Perak also contained an aboriginal (Orang Asli) settlement. A ~5% sub-sample from 40,736 chigger specimens was identified from five species of small mammals (n = 217), revealing 14 chigger species, including two new records for Malaysia. The abundance and species richness of chiggers were significantly affected by habitat type (highest in forest border), state (highest in Perak), and season (highest in dry). The overall prevalence of Orientia tsutsugamushi DNA in small-mammal tissues was 11.7% and was not significantly affected by host or habitat characteristics, but in Johor, was positively associated with infestation by Leptotrombidium arenicola. These findings highlight the risk of contracting scrub typhus in oil palm plantations and associated human settlements.