Microbiome and mitogenomics of the chigger mite Pentidionis agamae: potential role as an Orientia vector and associations with divergent clades of Wolbachia and Borrelia.

BACKGROUND: Trombiculid mites are globally distributed, highly diverse arachnids that largely lack molecular resources such as whole mitogenomes for the elucidation of taxonomic relationships. Trombiculid larvae (chiggers) parasitise vertebrates and can transmit bacteria (Orientia spp.) responsible for scrub typhus, a zoonotic febrile illness. Orientia tsutsugamushi causes most cases of scrub typhus and is endemic to the Asia-Pacific Region, where it is transmitted by Leptotrombidium spp. chiggers. However, in Dubai, Candidatus Orientia chuto was isolated from a case of scrub typhus and is also known to circulate among rodents in Saudi Arabia and Kenya, although its vectors remain poorly defined. In addition to Orientia, chiggers are often infected with other potential pathogens or arthropod-specific endosymbionts, but their significance for trombiculid biology and public health is unclear. RESULTS: Ten chigger species were collected from rodents in southwestern Saudi Arabia. Chiggers were pooled according to species and screened for Orientia DNA by PCR. Two species (Microtrombicula muhaylensis and Pentidionis agamae) produced positive results for the htrA gene, although Ca. Orientia chuto DNA was confirmed by Sanger sequencing only in P. agamae. Metagenomic sequencing of three pools of P. agamae provided evidence for two other bacterial associates: a spirochaete and a Wolbachia symbiont. Phylogenetic analysis of 16S rRNA and multi-locus sequence typing genes placed the spirochaete in a clade of micromammal-associated Borrelia spp. that are widely-distributed globally with no known vector. For the Wolbachia symbiont, a genome assembly was obtained that allowed phylogenetic localisation in a novel, divergent clade. Cytochrome c oxidase I (COI) barcodes for Saudi Arabian chiggers enabled comparisons with global chigger diversity, revealing several cases of discordance with classical taxonomy. Complete mitogenome assemblies were obtained for the three P. agamae pools and almost 50 SNPs were identified, despite a common geographic origin. CONCLUSIONS: P. agamae was identified as a potential vector of Ca. Orientia chuto on the Arabian Peninsula. The detection of an unusual Borrelia sp. and a divergent Wolbachia symbiont in P. agamae indicated links with chigger microbiomes in other parts of the world, while COI barcoding and mitogenomic analyses greatly extended our understanding of inter- and intraspecific relationships in trombiculid mites.

Molecular detection of Orientia tsutsugamushi in ectoparasites & their small mammal hosts captured from scrub typhus endemic areas in Madurai district, India.

BACKGROUND OBJECTIVES: Scrub typhus, caused by Orientia tsutsugamushi present in small mammals harbouring the ectoparasites. A study was undertaken to detect the pathogen present in small mammals and its ectoparasites in the scrub typhus-reported areas. METHODS: The small mammals (rodents/shrews) and its ectoparasites were screened for O. tsutsugamushi using nested PCR amplification of the groEL gene. Small mammals were collected by trapping and screened for ectoparasites (mites, ticks and fleas) by combing method. RESULTS: All the chigger mites collected were tested negative for O. tsutsugamushi . Interestingly, adult non-trombiculid mites ( Oribatida sp., Dermanyssus gallinae ), fleas ( Xenopsylla astia, X. cheopis, Ctenophalides felis and Ctenophalides sp.) and ticks ( Rhipicephalus sanguineus , R. haemaphysaloides ) screened were found to be positive for O. tsutsugamushi , which the authors believe is the first report on these species globally. Bandicota bengalensis with O. tsutsugamushi infection is reported for the first time in India. The O. tsutsugamushi groEL sequences from the positive samples were similar to the reference strains, Karp and Ikeda and phylogenetically clustered in clade IV with less evolutionary divergence. The blood samples of Rattus rattus , Suncus murinus and B. bengalensis collected from this area were tested positive for O. tsutsugamushi ; interestingly, the sequence similarity was much pronounced with their ectoparasites indicating the transmission of the pathogen to host or vice versa . INTERPRETATION CONCLUSIONS: The outcome of the present investigations widened our scope on the pathogens present in ectoparasites and rodents/shrews from this area. This will help to formulate the required vector control methods to combat zoonotic diseases.

Bat ectoparasites: chigger mites (Trombiculidae), ticks (Ixodidae and Argasidae), and bugs (Cimicidae) in the Eastern Palaearctic.

Nine species-level taxa of bat ectoparasites, three chigger mites (Trombiculidae), three hard (Ixodidae), and one soft tick (Argasidae) species, as well as two bug (Cimicidae) species from nine bat species hosts were detected in the Eastern Palaearctic. Trombiculid larvae of Leptotrombidium schlugerae, Leptotrombidium album, and Ascoschoengastia latyshevi were first recorded on bats in the temperate zone of eastern Russia. L. schlugerae was more abundant than A. latyshevi in the same study sites in Eastern Siberia, and the main hosts of both chigger species were Plecotus ognevi and Eptesicus nilssonii. Ixodid ticks Dermacentor marginatus, Ixodes simplex, and Ixodes sp. were sampled from bats in Kazakhstan, the Far East, and Eastern Siberia, respectively. Phylogenetic analysis based on Cox1, 16S rDNA, and ITS2 sequences of I. simplex showed that the specimens from the Far East grouped into a clade distributed in the Eastern Palaearctic and India. In turn, the specimen of Ixodes sp. from Eastern Siberia was most closely related to Ixodes soricis and Ixodes angustus with p-distance of 9.8-10.7% (Cox1), suggesting that this tick probably belongs to a new species. Argas vespertilionis larvae were collected from three widespread bat species in Kazakhstan. Two bug species, Cimex pipistrelli and Cimex aff. lectularius, were recorded in the Far East and Eastern Siberia, respectively. Specimens from Transbaikalia were morphologically identified as Cimex lectularius. However, they differed from the latter by 12.5-12.9% of Cox1 sequences, indicating that C. aff. lectularius may be a new species.

Resistance of dwarf Cocos nucifera L. (Arecaceae) cultivars to Raoiella indica Hirst (Acari: Tenuipalpidae).

The red palm mite Raoiella indica Hirst, 1924 (Acari: Tenuipalpidae) is an important pest of the coconut palm Cocos nucifera L. (Arecaceae) and has caused problems in coconut production worldwide. Research has been carried out aiming at controlling the mite through chemical, biological, alternative, and host plant resistance methods. Identifying coconut palm cultivars resistant to R. indica is important to reduce the problems caused to plantations. Therefore, the objective of this work was to evaluate the performance of R. indica in six dwarf coconut palm cultivars, to identify sources of resistance. The cultivars of the sub-varieties green, red, and yellow evaluated were Brazilian Green Dwarf-Jiqui (BGDJ), Brazilian Red Dwarf-Gramame (BRDG), Cameroon Red Dwarf (CRD), Malayan Red Dwarf (MRD), Brazilian Yellow Dwarf-Gramame (BYDG), and Malayan Yellow Dwarf (MYD). Confinement and free choice tests of R. indica on the cultivars were performed, in which biological parameters and preference were evaluated. Mite performance was different in the cultivars evaluated. In the confinement bioassay, R. indica had the worst performance in the cultivar BGDJ, the best performance in CRD, MRD, and BRDG, and intermediate performance in BYDG and MYD. In the free choice test, the cultivars MRD and MYD were preferred in relation to BGDJ, and CRD was less preferred in relation to BGDJ. Therefore, we considered that the cultivar BGDJ is the most resistant to R. indica, by antibiosis and antixenosis; CRD has resistance by antixenosis; and MRD, BRDG, BYDG, and MYD are susceptible.

Parasite infestation patterns differ between ticks and chigger mites on two rodent host species in Taiwan.

Parasites are typically concentrated on a few host individuals, and identifying the mechanisms underlying aggregated distribution can facilitate a more targeted control of parasites. We investigated the infestation patterns of hard ticks and chigger mites on two rodent species, the striped field mouse, Apodemus agrarius, and the lesser ricefield rat, Rattus losea, in Taiwan. We also explored abiotic and biotic factors that were important in explaining variation in the abundance of ticks and chiggers on rodent hosts. Ticks were more aggregated than chiggers on both rodent species. Factors important for the variation in parasitic loads, especially biotic factors, largely differed between ticks and chiggers. Variation partitioning analyses revealed that a larger proportion of variation in chiggers than in ticks can be explained, especially by abiotic factors. If, as proposed, the higher number of parasites in males is due to a larger range area or immunity being suppressed by testosterone, when A. agrarius males host more ticks, they are expected to also host more chiggers, given that chiggers adopt a similar host finding approach to that of ticks. Instead, the similar abundance of chiggers in male and female A. agrarius implies that a large home range or suppressed immunity does not predispose males to inevitably host more parasites. More variations were explained by abiotic than biotic factors, suggesting that controlling practices are more likely to be successful by focusing on factors related to the environment instead of host traits. Our study indicated that the extent of parasitism is rarely determined by a sole factor, but is an outcome of complex interactions among animal physiology, animal behavior, characteristics of parasites, and the environments.

Detection of Orientia spp. Bacteria in Field-Collected Free-Living Eutrombicula Chigger Mites, United States.

Scrub typhus, a rickettsial disease caused by Orientia spp., is transmitted by infected larval trombiculid mites (chiggers). We report the molecular detection of Orientia species in free-living Eutrombicula chiggers collected in an area in North Carolina, USA, to which spotted fever group rickettsiae infections are endemic.

Molecular Detection of Candidatus Orientia chuto in Wildlife, Saudi Arabia.

Scrub typhus is a zoonosis caused by 3 species of Orientia bacteria, including Candidatus Orientia chuto. This species is known only from a human case in Dubai and infections in wildlife in Kenya. We report molecular detection of Candidatus O. chuto in 2 wild rodent species from Saudi Arabia.

Bacterial pathogens' screening in Brazilian chigger mites (Trombidiformes: Trombiculidae), with the first report of 'Candidatus Rickettsia colombianensi'-like in avian-associated chiggers.

Chiggers are larval ectoparasites of the Trombiculidae that can transmit pathogens to their hosts. In this study, chiggers collected from birds in Brazil were morphologically identified as Blankaartia sinnamaryi, Eutrombicula batatas, Eutrombicula daemoni, Eutrombicula goeldii, Eutrombicula tinami, and Parasecia gilbertoi. For these specimens, a beginning attempt at molecular identification were also provided, as well as, were genetically screened to detect bacterial pathogens. The species B. sinnamaryi and E. tinami were positive for Rickettsia felis-like and 'Candidatus Rickettsia colombianensi'-like, respectively. For the other agents (Anaplasmataceae, Borrelia spp. and Orientia tsutsugamushi), the tests were negative. This is the first report of 'Ca. R. colombianensi'-like and the second record of R. felis-like in chigger collected on birds from Brazil.

Ectoparasites associated with the Bushveld gerbil (Gerbilliscus leucogaster) and the role of the host and habitat in shaping ectoparasite diversity and infestations.

Rodents are known hosts for various ectoparasite taxa such as fleas, lice, ticks and mites. South Africa is recognized for its animal diversity, yet little is published about the parasite diversity associated with wild rodent species. By focusing on a wildlife-human/domestic animal interface, the study aims to record ectoparasite diversity and levels of infestations of the Bushveld gerbil, Gerbilliscus leucogaster, and to establish the relationship between ectoparasite infestation parameters and host- and habitat factors. Rodents (n = 127) were trapped in 2 habitat types (natural and agricultural) during 2014­2020. More than 6500 individuals of 32 epifaunistic species represented by 21 genera and belonging to 5 taxonomic groups (fleas, sucking lice, ticks, mesostigmatan mites and trombiculid mites) were collected. Mesostigmatan mites and lice were the most abundant and fleas and mesostigmatan mites the most prevalent groups. Flea and mesostigmatan mite numbers and mesostigmatan mite species richness was significantly higher on reproductively active male than female rodents. Only ticks were significantly associated with habitat type, with significantly higher tick numbers and more tick species on rodents in the natural compared to the agricultural habitat. We conclude that the level of infestation by ectoparasites closely associated with the host (fleas and mites) was affected by host-associated factors, while infestation by ectoparasite that spend most of their life in the external environment (ticks) was affected by habitat type.

Chigger mite (Acariformes: Trombiculidae) infestation in reed passerine birds in Central Europe: a case of the bearded tit Panurus biarmicus.

Larval trombiculid (chigger) mites are common ectoparasites of terrestrial vertebrates including humans, causing itching and skin inflammation known as trombiculiasis. Investigation of their diversity, distribution and seasonal abundance is therefore important from a veterinary and public health point of view. Although researchers have paid increased attention to these parasites in recent years, there is still little ecological data available on chiggers associated with birds inhabiting different types of habitats such as wetlands, for example. In 2021, we investigated the mite fauna in a specialist reedbed passerine, the bearded tit (Panurus biarmicus), and their effects on this host in the south-west Slovakia, Central Europe. A total of 1134 larvae of 1 mite species Blankaartia acuscutellaris were found in 99 out of 267 examined bearded tits. Juveniles were more infested than adult birds, but no differences were found between sexes. The larvae of mites first appeared on the host during the second half of June and peaked in the second half of July. After that, their numbers decreased gradually until October. Despite the relatively high prevalence and intensity of mite infestation in the bearded tit, no differences in body condition between infested and uninfested birds suggest that infestation by B. acuscutellaris may not have serious negative effects on the host health. Bearded tits can therefore be a reliable indicator of the presence of the chigger mites in wetland habitats.

Microbiome comparison of Dermanyssus gallinae populations from different farm rearing systems and the presence of common endosymbiotic bacteria at developmental stages.

The hematophagous arthropod, Dermanyssus gallinae (Poultry red mite, PRM) can cause remarkable economic losses in the poultry industry across the globe. Although overall composition of endosymbiotic bacteria has been shown in previous studies, how farm habitats influence the microbiome remains unclear. In the present study, we compared the bacterial communities of D. gallinae populations collected from the cage and free-range farms using next-generation sequences targeting the V3-V4 hypervariable region of the 16S rRNA gene. The QIIME2 pipeline was followed in bioinformatic analyses. Proteobacteria represented a great majority of the total bacterial community of D. gallinae from both farming systems. More specifically, Bartonella-like bacteria (40.8%) and Candidatus Cardinium (21.5%) were found to be predominant genera in free-range and cage rearing systems, respectively. However, the microbiome variation based on farming systems was not statistically significant. In addition, the presence of the five common endosymbiotic bacteria (Wolbachia, Cardinium, Rickettsiella, Spiroplasma, and Schineria) was screened in different developmental stages of D. gallinae. Cardinium was detected in all developmental stages of D. gallinae. On the other hand, Wolbachia and Rickettsiella were only found in adults/nymphs, but neither in the eggs nor larvae. To our knowledge, this study provides the first microbiome comparison at genus-level in D. gallinae populations collected from different farm habitats and will contribute to the knowledge of the biology of D. gallinae.

Successful long-term control of poultry red mite (Dermanyssus gallinae) infestations in floor-kept laying hens via integrated pest management-a case report.

This case report describes the successful control of poultry red mite [PRM] (Dermanyssus gallinae) infestations in an experimental laying hen house via a combined use of cleaning and disinfection measure, the preventive application of a synthetic silica-based acaricide and frequent mite monitoring. The high number of PRM in the laying hen house was reduced by 99.8% by treatment with fluralaner (Exzolt®, MSD Animal Health Unterschleißheim, Germany; 0.5 mg/kg body weight via drinking water twice, 7 days apart). After the laying hens were removed, the hen house was dry-cleaned, wet-cleaned and disinfected. After drying, synthetic amorphous silica (Fossil Shield® instant white, Bein GmbH, Eiterfeld, Germany) was applied as a preventive measure before the hen house was restocked with pullets for two housing periods of 58 and 52 weeks. Over these periods (i.e. more than 2 years), no PRM was detected during mite monitoring at two-week intervals via tube traps and visual monitoring. This result therefore suggests that the combined use of appropriate chemical and physical prevention measures within an integrated pest management regime can be successfully used for the long-term control of PRM. This could reduce the use of acaricidal drugs, thereby helping maintain their efficacy.

Current status of mites and mite-borne diseases in India.

Mites act as the vectors of zoonotic vector-borne disease scrub typhus caused by the rickettsial pathogen Orientia tsutsugamushi. In India, scrub typhus is transmitted by the mite Leptotrombidium deliense. Rodents are the reservoirs and support the spread of this disease. Sarcoptes scabiei or the itch mite is causing scabies which is a common skin infection in India. Dermatitis, trombiculosis, and mite dust allergy are common mite-borne diseases transmitted by Pyemotidae family. D. brevis folliculorum and Demodex brevis are two major mite species found on humans also causing a disease; "Demodicosis", common in India. Dermanyssus gallinae, fowl mite causes pruritis in poultry birds, transmits infections to poultry workers when they come into contact with birds. There is a re-emergence of mite-borne diseases, especially scrub typhus, in many parts of India requiring urgent attention for its control. This review is aimed to update the available information on mites and mite transmitted diseases prevalent in India to highlight the importance of rodent and chigger mite vector control to prevent forthcoming mite-borne diseases in India.

Let me know your name: a study of chigger mites (Acariformes: Trombiculidae) associated with the edible dormouse (Glis glis) in the Carpathian-Balkan distribution gradient.

Trombiculid mites were collected from the edible dormouse (Glis glis) within the Carpathian-Balkan distribution gradient of host species. Representatives of five genera (Leptotrombidium, Neotrombicula, Brunehaldia, Hirsutiella, Schoutedenichia) and 10 species of chiggers were discovered in the material, based on morphological and/or molecular data. Brunehaldia, new to the fauna of Greece, was recorded for the first time from the edible dormouse. Neotrombicula talmiensis was new to the fauna of Greece and Neotrombicula vulgaris was new to the fauna of North Macedonia. Successful amplification and sequencing of COI was carried out in relation to three genera and six species. The intraspecific variation of taxa hitherto distinguished based on morphological criteria was juxtaposed with molecular data, using the distance method and the phylogenetic approach. The molecular methods indicated wider than hitherto recognized, intraspecific morphological variation for Leptotrombidium europaeum and N. talmiensis. On the other hand, an inference limited to morphology proved to be insufficient for species delineation, which was confirmed by the relatively low identity (%) of examined COI sequences as well as the size of inter-/intraspecific K2P distance threshold. Our study provides support for integrative taxonomy that combines different sources of evidence and contributes to recognition of the scope of intraspecific variation. The high degree of hidden diversity revealed with the application of molecular tools, votes for a careful approach to the identification of chiggers. The confirmed cases of co-invasion, including the representatives of various genera (Leptotrombidium and Neotrombicula, Brunehaldia and Neotrombicula, Neotrombicula and Schoutedenichia, Hirsutiella and Schoutedenichia) additionally support the need to include all larvae found on a given host specimen in the identification process.

Species diversity and related ecology of chiggers on small mammals in a unique geographical area of Yunnan Province, southwest China.

Chiggers are common ectoparasites and the exclusive vector of scrub typhus. Based on previous investigations from a unique geographical area in Yunnan Province of southwest China, the Three Parallel Rivers Area, we retrospectively studied the species diversity and related ecology of chiggers on rodents and other small mammals. A very high species diversity of 120 chigger species was identified. Five dominant chigger species accounted for 59.4% (5238/8965) of total chiggers, and among them Leptotrombidium scutellare is the second major vector of scrub typhus in China. Species diversity of the chigger community fluctuates greatly in different altitudinal and latitudinal gradients. There are significant differences in species composition, species diversity and dominant species of chiggers among hosts with apparent community heterogeneity. Based on the species abundance distribution, the expected total number of chigger species was estimated to be 170, 50 more than the number of actually collected species; this further indicates a very high chigger species diversity in this area. The bipartite ecological network analysis revealed the intricate relationships between chigger and host species-positive and negative correlations existed among some species of dominant and vector chiggers.

A new species of Eutrombicula Ewing, 1938 (Trombidiformes: Trombiculidae) from the Galápagos Islands, with a checklist of Ecuadorian chiggers.

Few studies have been carried out on the Ecuadorian chigger fauna. In this study, we are describing a new species of Eutrombicula from the Galápagos Islands. Also, we provide an updated checklist of Ecuadorian and Galápagos Islands chiggers, including a new host association and locality record for Eutrombicula nerudai Stekolnikov and González-Acuña.

Novel Vector of Scrub Typhus in Sub-Antarctic Chile: Evidence From Human Exposure.

The exposure of a research team to chigger mites in southern Chile allowed the first identification of a trombiculid species as vector and reservoir of scrub typhus outside the tsutsugamushi triangle, providing unique insights into the ecology and transmission of this recently discovered rickettsial infection in South America.

Mixed genotypes of Orientia tsutsugamushi in conserved genes and a single immune-dominant tsa56 genotype discovered from a patient with scrub typhus in Hainan Island, China: a case report.

BACKGROUND: Orientia tsutsugamushi (O. tsutsugamushi), an obligate intracellular bacterium, is transmitted to humans through infected larval trombiculid mite bites, causing scrub typhus. Mixed genotypes of O. tsutsugamushi in canonical conserved genes were reported in 8-25% of blood samples from patients. Yet, there are few clinical descriptions of these mixed O. tsutsugamushi-infected patients. CASE PRESENTATION: We report a patient with scrub typhus complicated with pulmonary involvement and hepatic dysfunction, who carried mixed genotypes of the conserved genes but had a single immune-dominant 56-kDa type-specific antigen (tsa56) genotype. The patient was successfully recovered by doxycycline treatment. CONCLUSIONS: In this reported case, both patient's eschar and blood samples have repeatedly shown the same results, i.e., no variants were discovered in tsa56 gene that bears multiple hypervariable regions. Whereas the selected highly conserved genes were identified with up to 32 variants in a 2700 base-pair concatenated sequence. The prevalence, disease severity and mechanism of these single-tsa56-genotype mixed infections remain to be investigated on a large scale with more cases.

Diversity and distribution of ectoparasite taxa associated with Micaelamys namaquensis (Rodentia: Muridae), an opportunistic commensal rodent species in South Africa.

South Africa boasts a rich diversity of small mammals of which several are commensal and harbour parasites of zoonotic importance. However, limited information is available on the parasite diversity and distribution associated with rodents in South Africa. This is particularly relevant for Micaelamys namaquensis (Namaqua rock mouse), a regionally widespread and locally abundant species that is often commensal. To address the paucity of data, the aims of the study were to record the ectoparasite diversity associated with M. namaquensis and develop distribution maps of lice and mites associated with M. namaquensis and other rodents in South Africa. Micaelamys namaquensis individuals (n = 216) were obtained from 12 localities representing multiple biomes during 2017­2018. A total of 5591 ectoparasites representing 5 taxonomic groups ­ fleas, lice, mesostigmatid mites, chiggers and ticks was recorded. These consisted of at least 57 taxa of which ticks were the most speciose (20 taxa). Novel contributions include new host and locality data for several ectoparasite taxa and undescribed chigger species. Known vector species were recorded which included fleas (Ctenocephalides felis, Dinopsyllus ellobius and Xenopsylla brasiliensis) and ticks (Haemaphysalis elliptica, Rhipicephalus appendiculatus and Rhipicephalus simus). Locality records indicate within-taxon geographic differences between the 2 louse species and the 2 most abundant mite species. It is clear that M. namaquensis hosts a rich diversity of ectoparasite taxa and, as such, is an important rodent species to monitor in habitats where it occurs in close proximity to humans and domestic animals.

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.