Euschoengastia pipistrelli (Acari: Trombiculidae) from American Perimyotis, Perimyotis subflavus (Chiroptera: Vespertilionidae): Novel Stereoscopic and Scanning Electron Microscopy.

The chigger, Euschoengastia pipistrelli Brennan, is a trombiculid mite that infests a variety of vespertilionid bats in North America. It has been reported from at least 9 species of bats from 18 U.S. states. However, nothing is available on the actual in situ infestation and ultrastructure of this chigger. Here we document some stereoscopic photographs of the infestation as well as a scanning electron micrograph of the mite from a common bat species. We also provide a summation of host and state records for this chigger.

Four Species of Under-reported Parasitic Arthropods in Mexico and their Potential Role as Vectors of Pathogens.

Numerous arthropod taxa are important in human and veterinary medicine. The salivary secretions and feces of arthropods can cause allergic reactions in host vertebrates or harbor pathogens. Also, bites can be a risk factor for secondary infections. Documenting the diversity of arthropods of medical and veterinary importance remains an important aspect of disease control and prevention. We provide new records of ectoparasitic arthropods from Mexico that are of potential medical or veterinary relevance. Scanning electron microscopy along with amplification and sequencing of a fragment of the mitochondrial gene (16S rRNA) was used to confirm some species identities. We report the cat louse Felicola subrostratus from cats and the chewing louse Heterodoxus spiniger from dogs, which are common ectoparasites but largely not reported in Mexico. The chigger Eutrombicula alfreddugesi is common on wild lizards (Squamata). For the first time, E. alfreddugesi is reported on Hemidactylus frenatus (common house gecko). This reptile has a close relationship with humans and its chiggers can cause dermatitis (i.e., trombiculiasis) or transmit pathogens. In addition, the common bed bug Cimex lectularius is reported for the first time in the state of Yucatan, an atypical area for its natural distribution. Phylogenetic analysis revealed that Cimex lectularius from Yucatan is closely related to genetic sequences of Cimex lectularius from China. Knowing the regional distribution of arthropods allows the design and implementation of prevention strategies for those that have potential roles as reservoirs or vectors.

A New Species of Morelacarus (Acariformes: Prostigmata: Leeuwenhoekiidae) Associated With Sceloporus grammicus (Squamata: Phrynosomatidae) From the Mexican Plateau, Zacatecas, Mexico.

A new species of chigger mites, Morelacarus uazi n. sp., parasitizing lizards of the species Sceloporus grammicus from Mazapil, Zacatecas, Mexico, is described and illustrated. This new taxon differs from other species of the genus by having a palpal claw with 4 subequal paired prongs, and it can be further distinguished by the combination of the following characters: presence of onychotriches and mastitarsala III, absence of mastitibiala III, subterminala I, and parasubterminala I, and the palpal setal formula B/B/BBb. Additionally, an identification key for the species of the genus Morelacarus is presented. The new species represents the seventh species assigned to the genus and the first species described from a Mexican lizard.

Autofluorescence microscopy for paired-matched morphological and molecular identification of individual chigger mites (Acari: Trombiculidae), the vectors of scrub typhus.

BACKGROUND: 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. METHODOLOGY/PRINCIPAL FINDINGS: 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. CONCLUSIONS/SIGNIFICANCE: 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.

Comparative ultrastructure of coxal glands in unfed larvae of Leptotrombidium orientale (Schluger, 1948) (Trombiculidae) and Hydryphantes ruber (de Geer, 1778) (Hydryphantidae).

Coxal glands of unfed larvae Leptotrombidium orientale (Schluger, 1948) (Trombiculidae), a terrestrial mite parasitizing vertebrates, and Hydryphantes ruber (de Geer, 1778) (Hydryphantidae), a water mite parasitizing insects were studied using transmission electron microscopy. In both species, the coxal glands are represented by a paired tubular organ extending on the sides of the brain from the mouthparts to the frontal midgut wall and are formed of the cells arranged around the central lumen. As in other Parasitengona, the coxal glands are devoid of a proximal sacculus. The excretory duct, joining with ducts of the prosomal salivary glands constitutes the common podocephalic duct, opening into the subcheliceral space. The coxal glands of L. orientale are composed of a distal tubule with a basal labyrinth, an intermediate segment without labyrinth, and a proximal tubule bearing tight microvilli on the apical cell surface and coiled around the intermediate segment. The coxal glands of H. ruber mainly consist of the uniformly organized proximal tubule with apical microvilli of the cells lacking the basal labyrinth. This tubule shows several loops running backward and forward in a vertical plane on the side of the brain. In contrast to L. orientale, larvae of H. ruber reveal a terminal cuticular sac/bladder for accumulation of secreted fluids. Organization of the coxal glands depends on the ecological conditions of mites. Larvae of terrestrial L. orientale possess distal tubule functioning in re-absorption of ions and water. Conversely, water mite larvae H. ruber need to evacuate of the water excess, so the filtrating proximal tubule is prominent.

Comparative stylostome ultrastructure of Hirsutiella zachvatkini (Trombiculidae) and Trombidium holosericeum (Trombidiidae) larvae.

Stylostomes (feeding tubes) of Hirsutiella zachvatkini (Schluger) (Trombiculidae), feeding on bank voles [Myodes glareolus (Schreber)], and of Trombidium holosericeum (L.) (Trombidiidae), feeding on larvae of Stenodemini sp. (Heteroptera, Miridae), were studied by TEM methods and on semi-thin sections. The stylostome of H. zachvatkini is a homogeneous structure of low electron density and without strict margins. It extends within the concave host epidermis, undergoing hyperplasia and hyperkeratosis. TEM does not reveal any obvious stratification in the stylostome walls. The cheliceral movable digits are moved apart by 5-6 µm and tightly applied/adhered to the stylostome substance. A local area beneath the open end of the stylostome canal is not empty but contains a nearly homogeneous substrate, which can pass into the central stylostome canal. The latter is mostly free of contents. In contrast to H. zachvatkini, larvae of T. holosericeum form a root-like stylostome chaotically branching within the clear space underneath the host cuticle free of tissue elements. Tubules of the distal stylostome branches become progressively thinner and disappear blindly. As in H. zachvatkini, the stylostome walls of T. holosericeum are devoid of stratification but show moderate to high electron density. The cheliceral movable digits are moved apart by the same distance, as in H. zachvatkini, and tightly applied to the stylostome substance. The lumen of the central canal is either electron lucent, in the distal portions, or filled with a fine granular or homogeneous substrate of low electron density in the proximal portions forming a type of ampoule. This study shows that Trombiculidae and Trombidiidae share similar initial stages of stylostome formation but the resultant stylostome of each family is distinctly different.

A new genus and species of chigger mite (Trombidiformes: Trombiculidae) from Loxodontomys pikumche (Rodentia: Cricetidae) in Chile.

The family Trombiculidae is one of the most diverse and cosmopolitan (Walter et al. 2009). In Chile, the family Trombiculidae is represented by six genera associated with reptiles: Eutrombicula Ewing; Microtrombicula Ewing; Paratrombicula Goff & Whitaker; Whartonacarus (Brennan & Jones); Diaguitacarus Stekolnikov & González-Acuña and Proschoengastia Vercammen-Grandjean and two genera associated with rodents Chilacarus Webb, Bennett & Loomis and Poliremotus Brennan & Goff (Stekolnikov & González-Acuña 2015).

Mouthparts in Leptotrombidium larvae (Acariformes: Trombiculidae).

Mouthparts of Leptotrombidium larvae (Acariformes: Trombiculidae), potential vectors of tsutsugamushi disease agents, were studied in detail using light microscopy, scanning electron microscopy, and transmission electron microscopy. The mouthparts incorporated within the pseudotagma gnathosoma are composed of the infracapitulum ventrally and the chelicerae dorsally. The ventral wall of the infracapitulum is formed by a wide mentum posteriorly and a narrowed malapophysis anteriorly. The malapophysis firmly envelops the distal cheliceral portions by its lateral walls. The lateral lips of the malapophysis are flexible structures hiding the cheliceral blades in inactive condition and turning back forming a type of temporary sucker closely applied to the host skin during feeding. The roof of the infracapitulum is formed by a weakly sclerotized labrum anteriorly and a cervix with the capitular apodemes extending posteriorly. The labral muscles are lacking. The capitular apodemes serve as origin for pharyngeal dilators running to the dorsal wall of the pharynx fused with the bottom of the infracapitulum. The basal cheliceral segments are separated from each other besides the very posterior portions where they are movably joined by the inner walls. The sigmoid pieces serve for insertion of the cheliceral elevators originating at the posterior portions of the basal segments. The movable digits reveal the solid basal sclerite and the cheliceral blade curved upward with a tricuspid cap on its tip. Dendrites of nerve cells run along the digits to their tips. The ganglia are placed within the basal segments just behind the movable digits. The chelicerae also reveal well developed flexible fixed digits overhanging the basal portions of the blades. The gnathosoma possesses several sets of extrinsic muscles originating at the scutum and at the soft cuticle behind it. Laterally, the gnathosoma bears five-segmented palps with a trifurcate palpal claw.

Comparative morphology and ultrastructure of the prosomal salivary glands in the unfed larvae Leptotrombidium orientale (Acariformes, Trombiculidae), a possible vector of tsutsugamushi disease agent.

The prosomal salivary glands of the unfed larvae Leptotrombidium orientale (Schluger) were investigated using transmission electron microscopy. In total, four pairs of the prosomal glands were identified--three pairs, the lateral, the medial and the anterior, belong to the podocephalic system, and one pair, the posterior, is separate having an own excretory duct. All glands are simple alveolar/acinous with prismatic cells arranged around a relatively small intra-alveolar lumen with the duct base. The cells of all glands besides the lateral ones contain practically mature electron-dense secretory granules ready to be discharged from the cells. The secretory granules in the lateral glands undergo formation and maturation due to the Golgi body activity. The cells of all gland types contain a large basally located nucleus and variously expressed rough endoplasmic reticulum. Specialized duct-forming cells filled with numerous freely scattered microtubules are situated in the middle zone of each gland's acinus and form the intra-alveolar lumen and the duct base. Both the acinar (secretory) and the duct-forming cells contact each other via gap junctions and septate desmosomes. Axons of nerve cells come close to the basal extensions of the duct-forming cells where they form the bulb-shaped synaptic terminations. The process of secretion is under the control of the nerve system that provides contraction of the duct-forming cells and discharge of secretion from the secretory cells into the intra-alveolar lumen and further to the exterior. Unfed larvae of L. orientale, the potential vector of tsutsugamushi disease agents, contain the most simply organized salivary secretory granules among known trombiculid larvae, and this secretion, besides the lateral glands, does not undergo significant additional maturation. Thus, the larvae are apparently ready to feed on the appropriate host just nearly after hatching.

Stylostome organization in feeding Leptotrombidium larvae (Acariformes: Trombiculidae).

The stylostome of larvae of the trombiculids Leptotrombidium scutellare (Nagayo et al.), Leptotrombidium fletcheri (Womersley et Heaslip) and Leptotrombidium deliense (Walch) was studied experimentally at different time intervals after larval attachment using the histological method. The stylostome of these species has the same organization and belongs to the epidermal combined with the mixed type, developing more in width than in length. Neither transverse nor conspicuous longitudinal layers are present within the stylostome walls, which stain predominantly in red with Azan, also showing longitudinal portions with blue staining. Larvae tend to attach closely to each other and scabs, consisting of the hyperkeratotic epidermal layers fusing with migrating inflammatory cells, develop around the attachment sites. The dermis shows inflammatory foci with dilated capillaries and inflammatory cells inserting in the connective tissue layer underneath the stylostome. The feeding cavity, which is moderately expressed, may be found either in the epidermis or in the dermis. It contains inflammatory cells and their debris in the liquefied host tissues. The stylostome length depends on the character of the attachment site (the thicker epidermis or scab the longer the stylostome), and does not directly correspond to the stages of larval feeding. Nevertheless, at the 48-h time interval, nearly all attached larvae are found to be fully fed and their midgut cells are filled with nutritional globules.

Severe ulceronecrotic dermatitis associated with mite infestation in the critically endangered Amargosa vole (Microtus californicus scirpensis).

The entire range of the critically endangered Amargosa vole (Microtus californicus scirpensis) consists of less than 20 km(2) of riparian habitat in the Amargosa River drainage of the Mojave Desert in southern California. In September 2010, deformities on ears and chiggers on the ears and genitalia were detected, with some individuals so severely affected that they were missing ear pinnae altogether. Follow-up trapping was performed to document the presence of mites and mite-associated disease, and molecular characterization was performed on the mites. Of 151 Amargosa voles sampled from February to April of 2011, 60 (39.7%) voles had hard orange mites adhered to some part of their bodies, on ears of 46 (76.7%), on genitalia of 11 (18.3%), and near mammary tissue of 13 (21.7%) voles. Gross lesions were not detected on genitalia, but 47% of all individuals examined showed pinnal lesions and deformities, which included alopecia, swelling, marginal necrosis, and ulceration, as well as scarring, scabbing, and loss of pinna mass covering 25-100% of the pinnae. Biopsies revealed parakeratotic hyperkeratosis and acanthosis with diffuse neutrophilic exocytosis and dense necrotic granulocytes in the epidermis and superficial dermis associated with focal erosion and ulceration. In the underlying dermis, there were dense pleocellular inflammatory cell infiltrates composed primarily of necrotic granulocytes and multifocal hemorrhage. In some samples, mite mouthparts could be seen penetrating the superficial epidermis associated with focal necrosis, and mite fragments were found on the surface epidermis and within hair follicles. Microscopic examination of the mites documented that they were a larval trombiculid in the genus Neotrombicula with anatomical features that most closely resemble Neotrombicula microti, based on scutal shape, setation, and texture. PCR of 2 mite pools (each consisting of 3 mites from an individual animal) amplified 331 bp amplicons, which had 92-97% homology with the 18S rRNA gene of Leptotrombidium deliense, although coverage of Trombiculidae in GenBank is sparse. The severity and prevalence of lesions due to this chigger were atypical and distinct. Severe clinical trombiculiasis in this endangered species could negatively impact individual health and fitness.

Trombiculid mites (Hannemania sp.) in Leptodactylus chaquensis (Amphibia: Anura) inhabiting selected soybean and rice agroecosystems of Argentina.

Trombiculid mites are known to parasitize a variety of amphibian species; however, few comparisons of mite parasitism among sites have been made. Here, Hannemania sp. parasitism in frogs (Leptodactylus chaquensis) inhabiting agroecosystems from mideastern Argentina was described. A total of 40 adult frogs (22 females and 18 males) were analyzed to detect ectoparasite Hannemania spp. larvae. Prevalence and mean abundance of Hannemania sp. were consistently higher in frogs from the agroecosystems (rice and soybean fields) than from two reference sites. Leptodactylus chaquensis might be considered an important host species of Hannemania sp., particularly in agricultural areas.

[Structure of eyes in trombidioid mites (Acariformes: Trombidioidea)]. / Stroenie glaz krasnotelkovykh kleshchei (Acariformes: Trombidioidea).

The eyes or ocelli of trombidioid mite larvae of Euschoengastia rotundata, Hirszutiella zachvatkini and Camerotrombidium pexatum, and larvae and adults of Platytrombidium fasciatum were studied by means of transmission electron microscopy. These species together with larvae of Odontacarus efferus, Ericotrombidium hasgelum, Walchia chinensis and adult E. rotundata and H. zachvatkini were also studied under scanning electron microscope. The eyes of larvae are not inverted and characterized by an epicuticular lamellar lens. The group of phoreceptor cells with rhabdomeres arranged typically of Chelicerata is underlaid by a pigment cup. The eyes of adult mites are inverted, perikarions of photoreceptor cells are situated between the lens and rhabdomeres; tapetum occupies the space between the pigment cup and rhabdomeres. Sensitivity of eyes to light is similar to that of primary eyes of spiders dwelling on soil surface.

Ultrastructure of the integument during moulting of the quiescent tritonymphal instar of trombiculid mite Hirsutiella zachvatkini (Acariformes: Trombiculidae).

The ultrastructure of the integument of the quiescent reduced tritonymph of the trombiculid mite Hirsutiella zachvatkini (Schluger) was investigated by means of transmission electron microscopy. Mites were investigated daily during the 14-16 day tritonymphal period (imagochrysalis). This period includes the deutonymphal moult (1-3 days), the quiescent tritonymph period (2-4 days), and the tritonymphal moult into the adult mite (6-10 days). A distinct recognizable feature of the tritonymphal moulting cycle is a sequence of events independent of precise time intervals. This process involves partial destruction and reorganization of the hypodermis of the previous instar, and formation of a new hypodermis of the subsequent instar from islands of rudimentary hypodermal cells. The integument of the reduced tritonymph differs greatly from that of both larva and active deutonymph and adult. It consists of a simply organized hypodermal layer of varying thickness and a thick clear poorly lamellate cuticle with curved pore canals, and lacking setae. The epicuticle is very thin and without a clear protein layer. The tritonymphal instar as such with its own cuticle situated near the hypodermis is encased within the detached covering of the previous active deutonymph, and may be considered a calyptostasic and entirely pharate instar. There is a tendency for reduced tritonymphal stage to be eliminated from ontogenesis and this stage is not homologous to the pupa of insects.

[Persistent scalp infestation by Dermanyssus gallinae in an Emilian country-woman]. / Infestazione persistente del cuoio capelluto da Dermanyssus gallinae in una contadina emiliana.

A case of persistent infestation of the scalp due to Dermanyssus gallinae.--Infestation due to Dermanyssus gallinae, the common red mite of poultry, in a country-woman aged 69 years from Crevalcore (Emilia-Romagna region, Northern Italy), is described. The case was unusual either for its location, the scalp, and for its persistence, 9 months. Specimens of the mite were also found in the henhouse adjacent to the patient's house. Apparently the woman recovered by means of daily washing of camomile tea.

A new species of Leptotrombidium (Acari:Trombiculidae) collected in active rice fields in northern Thailand.

Leptotrombidium (Leptotrombidium) chiangraiensis Tanskul & Linthicum is described and illustrated as new from specimens collected from the rodents Rattus rattus (L., 1758), Rattus argentiventer (Robinson & Kloss, 1916), Rattus losea (Swinhoe, 1870), and Bandicota indica (Bechstein, 1800) in Chiangrai Province northern Thailand. The new species was collected in active rice fields and adjacent fruit plantation areas. The etiological agent of scrub typhus, Orientia (formerly Rickettsia) tsutsugamushi (Hayashi), has been isolated from patients who live and work in the same habitat where L. chiangraiensis is the predominant Leptotrombidium species.

An ultrastructural study of vertical transmission of Rickettsia tsutsugamushi during oogenesis and spermatogenesis in Leptotrombidium pallidum.

Rickettsia tsutsugamushi in Leptotromibidium pallidum was observed by electron microscopy and rickettsiae were found in the various tissues and organs of both larvae and adults. Budding of rickettsiae, a manner of release from the host cells, was observed only in the rudiments of the reproductive organs in larvae. Oogonia and maturing oocytes in adult females and eggs after oviposition contained the microorganisms. In adult males, rickettsiae were also found in the spermatogonia, spermatocytes, and spermatids in the early stage of spermatogenesis, but were eliminated from these cells during maturation. Only the maturing spermatids, but not the eliminated rickettsiae, migrated to another rickettsia-free area of the testis, resulting in the separation of spermatids from rickettsiae and in the production of rickettsia-free spermatophores. Based on these observations, the mechanism of vertical transmission of the rickettsiae to the progeny occurs only in the female parents. Most rickettsiae in the somatic cells of larvae and adults were coccoid, but some rickettsiae in the ovary and the testis of adult mites showed bacillary forms and were enveloped by a membrane of unknown origin.

[The alveolar salivary glands of hungry larvae of the chigger Hirsutiella zachvatkini]. / Al'veoliarnye sliunnye zhelezy golodnykh lichinok krasnotelkovogo kleshcha Hirsutiella zachvatkini.

Heteromorphic parasitic larvae of Hirsutiella zachvatkini have four pairs of simple alveolar salivary glands possessing characteristic peculiarities of electron microscopic organisation. Each of paired granulosecreting salivary glands has its own dynamics of secretory activity, which is stimulated by a specific functional role connected with feeding on the host.

Electron microscopic observations of the embryo Leptotrombidium (Leptotrombidium) pallidum naturally infected with Rickettsia tsutsugamushi.

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.