Ubiquity and Diversity of Human-Associated Demodex Mites

Demodex mites are a group of hair follicle and sebaceous gland-dwelling species. The species of these mites found on humans are arguably the animals with which we have the most intimate interactions. Yet, their prevalence and diversity have been poorly explored. Here we use a new molecular method to assess the occurrence of Demodex mites on humans. In addition, we use the 18S rRNA gene (18S rDNA) to assess the genetic diversity and evolutionary history of Demodex lineages. Within our samples, 100% of people over 18 years of age appear to host at least one Demodex species, suggesting that Demodex mites may be universal associates of adult humans. A phylogenetic analysis of 18S rDNA reveals intraspecific structure within one of the two named human-associated Demodex species, D. brevis. The D. brevis clade is geographically structured, suggesting that new lineages are likely to be discovered as humans from additional geographic regions are sampled.     

Detection,Prevalence and Phylogenetic Relationships of Demodex spp and further Skin Prostigmata Mites (Acari,Arachnida) in Wild and Domestic Mammals

This study was conceived to detect skin mites in social mammals through real-time qPCR, and to estimate taxonomic Demodex and further Prostigmata mite relationships in different host species by comparing sequences from two genes: mitochondrial 16S rRNA and nuclear 18S rRNA. We determined the mite prevalence in the hair follicles of marmots (13%) and bats (17%). The high prevalence found in marmots and bats by sampling only one site on the body may indicate that mites are common inhabitants of their skin. Since we found three different mites (Neuchelacheles sp, Myobia sp and Penthaleus sp) in three bat species (Miotis yumanensis, Miotis californicus and Corynorhinus townsendii) and two different mites (both inferred to be members of the Prostigmata order) in one marmot species (Marmota flaviventris), we tentatively concluded that these skin mites 1) cannot be assigned to the same genus based only on a common host, and 2) seem to evolve according to the specific habitat and/or specific hair and sebaceous gland of the mammalian host. Moreover, two M. yumanensis bats harbored identical Neuchelacheles mites, indicating the possibility of interspecific cross-infection within a colony. However, some skin mites species are less restricted by host species than previously thought. Specifically, Demodex canis seems to be more transmissible across species than other skin mites. D. canis have been found mostly in dogs but also in cats and captive bats. In addition, we report the first case of D. canis infestation in a domestic ferret (Mustela putorius). All these mammalian hosts are related to human activities, and D. canis evolution may be a consequence of this relationship. The monophyletic Demodex clade showing closely related dog and human Demodex sequences also supports this likely hypothesis.     

Complete mitochondrial genomes of the human follicle mites Demodex brevis and D. folliculorum: novel gene arrangement,truncated tRNA genes,and ancient divergence between species

Background

Follicle mites of the genus Demodex are found on a wide diversity of mammals, including humans; surprisingly little is known, however, about the evolution of this association. Additional sequence information promises to facilitate studies of Demodex variation within and between host species. Here we report the complete mitochondrial genome sequences of two species of Demodex known to live on humans—Demodex brevis and D. folliculorum—which are the first such genomes available for any member of the genus. We analyzed these sequences to gain insight into the evolution of mitochondrial genomes within the Acariformes. We also used relaxed molecular clock analyses, based on alignments of mitochondrial proteins, to estimate the time of divergence between these two species.

Results

Both Demodex genomes shared a novel gene order that differs substantially from the ancestral chelicerate pattern, with transfer RNA (tRNA) genes apparently having moved much more often than other genes. Mitochondrial tRNA genes of both species were unusually short, with most of them unable to encode tRNAs that could fold into the canonical cloverleaf structure; indeed, several examples lacked both D- and T-arms. Finally, the high level of sequence divergence observed between these species suggests that these two lineages last shared a common ancestor no more recently than about 87 mya.

Conclusions

Among Acariformes, rearrangements involving tRNA genes tend to occur much more often than those involving other genes. The truncated tRNA genes observed in both Demodex species would seem to require the evolution of extensive tRNA editing capabilities and/or coevolved interacting factors. The molecular machinery necessary for these unusual tRNAs to function might provide an avenue for developing treatments of skin disorders caused by Demodex. The deep divergence time estimated between these two species sets a lower bound on the time that Demodex have been coevolving with their mammalian hosts, and supports the hypothesis that there was an early split within the genus Demodex into species that dwell in different skin microhabitats.

Electronic supplementary material

The online version of this article (doi:10.1186/1471-2164-15-1124) contains supplementary material, which is available to authorized users.     

The weasel (Mustela nivalis) helps in detectingDemodex mites in rodents

Many stages of parasites living in or on prey animals can pass undestroyed through the intestinal tract of carnivores and carnivorous animals. Nematode eggs, whole nematodes, even cestodes can be found in their faeces, but of the ectoparasitic mites usually only the eggs are found. It was therefore surprising thatDemodex mites, some of them even still alive, were found to have been passed by a captive weasel.Members of the genusDemodex cannot be detected by routine procedures yielding ectoparasites. Most species do not give rise to pathological alterations and the site they inhabit may not be known. To isolate them, either the skin complex or the organ supposed to containDemodex has to be digested with KOH or else the mites have to be squeezed out of the follicles. Compared to this, the weasel method is very simple.Faecal pellets (not more than one or two) are carefully ground with a mortar in a saturated ZnSO₄ solution and poured through a fine mesh into a test tube so as to form a little mountain at the orifice. The tube is allowed to stand for 25–30 min. Due to their lower specific weight, protozoan oocysts, worm and mite eggs and the specimens ofDemodex rise to the surface. The meniscus is then touched with a cover slip and, with the adhering drop, is placed on a slide. In the case of the parasites mentioned here, the procedure can be repeated once more after 5 min. Demodex mites and all their developmental stages, including the eggs, appear in a clean preparation on the slide, ready to be measured, drawn, or photographed. The mites are in excellent shape. Some of them even move their gnathosome and legs, if fresh faeces are submitted to the concentration technique; the method is very quick, with results being obtained within 30 min. Although only one rodent can be fed to a weasel per day, it takes a minimum of labour to examine the faeces, which can also be stored in a refrigerator.As the weasel was young when caught, it was not restricted in its feeding habits to the common vole (Microtus arvalis) which is its main prey in Middle Europe, but took other rodents, too. This proved to be very helpful when two different rodent species were to be tested forDemodex. In order not to mix them up, a white mouse of aDemodex-free stock was given on the day between. By this it was also possible to make a semi-quantitative survey of theDemodex population size of one host animal, because the faecal pellets of a weasel are very small and can be entirely used. All available rodents can be tested by this method. If the weasel is hungry it might even be possible to feed only the parts of the body of bigger host animals suspected to containDemodex.The disadvantages of the weasel method are that (a) no permanent mounts can be made, because the zinc sulfate (or any other saturated solution commonly used for these purposes) forms crystals when a mounting medium is added, and (b) the site inhabited by the mites has still to be ascertained; this is particularly important, since one host species can harbour more than oneDemodex species, and these may look very similar.     

Efficient isolation and observation of the most complex human commensal, <Emphasis Type="Italic">Demodex</Emphasis> spp.

Demodex spp. mites are an often neglected member of the human skin microbiome. Mostly they are commensals, although their pathophysiological role in rosacea, spinulosis folliculorum, and other skin diseases is recognized. Little is known about their life cycle, biology, and physiology. Demodex mites cannot be cultivated in vitro, thereby complicating research immensely. The manual extraction from human sebum is laborious and death can only be detected by surrogate markers like ceased movement or loss of fluorescence. Here we present a new approach for the extraction of larger mite numbers and the hitherto most precise way to detect death. The extraction of mites from sebum and debris by hand can be accelerated by a factor 10 using sucrose gradient centrifugation, which is well tolerated by the mites. Staining with propidium iodide allows for easy identification of dead mites, excluding frail mites that stopped moving, and has no negative effect on overall mite survival. We anticipate our methods to be a starting point for more sophisticated research and ultimately in vitro cultivation of Demodex spp. mites.     

Tear cytokines and chemokines in patients with Demodex blepharitis

PurposeThe purpose of the study is to evaluate the causes of inflammation in Demodex-induced blepharitis by analyzing cytokine levels in lacrimal fluid.MethodsFifteen Demodex blepharitis patients were selected for assessment of tear cytokine concentrations. Fifteen Demodex-free blepharitis patients and 15 subjects with no ocular symptoms were selected as control groups. Minimally stimulated tear samples (20 μl) were collected from each eye and analyzed using a Luminex® 200? Total System for detection of IL-1β, IL-5, IL-7, IL-12, IL-13, IL-17, granulocyte colony-stimulating factor (G-CSF), and macrophage inflammatory protein-1 beta (MIP-1β).ResultsThe concentration of IL-17 in tears was significantly higher in the Demodex blepharitis group than in the Demodex-free blepharitis group. Tear IL-7 and IL-12 levels show serial increases for these three groups (p < 0.05). There were no significant differences in the other cytokines levels between both blepharitis groups. We confirmed that elevated cytokines normalized after treatments.ConclusionsInfestation of Demodex mites induces change of tear cytokine levels, IL-17 especially, which cause inflammation of the lid margin and ocular surface. These findings might increase our understanding of the mechanism of ocular discomfort and telangiectasias frequently found in Demodex blepharitis patients.     

Genetic and morphological diversity of Trisetacus species (Eriophyoidea: Phytoptidae) associated with coniferous trees in Poland: phylogeny,barcoding, host and habitat specialization

Eriophyoid species belonging to the genus Trisetacus are economically important as pests of conifers. A narrow host specialization to conifers and some unique morphological characteristics have made these mites interesting subjects for scientific inquiry. In this study, we assessed morphological and genetic variation of seven Trisetacus species originating from six coniferous hosts in Poland by morphometric analysis and molecular sequencing of the mitochondrial cytochrome oxidase subunit I gene and the nuclear D2 region of 28S rDNA. The results confirmed the monophyly of the genus Trisetacus as well as the monophyly of five of the seven species studied. Both DNA sequences were effective in discriminating between six of the seven species tested. Host-dependent genetic and morphological variation in T. silvestris and T. relocatus, and habitat-dependent genetic and morphological variation in T. juniperinus were detected, suggesting the existence of races or even distinct species within these Trisetacus taxa. This is the first molecular phylogenetic analysis of the Trisetacus species. The findings presented here will stimulate further investigations on the evolutionary relationships of Trisetacus as well as the entire Phytoptidae family.     

Influence of temperature and medium on viability of <Emphasis Type="Italic">Demodex folliculorum</Emphasis> and <Emphasis Type="Italic">Demodex brevis</Emphasis> (Acari: Demodicidae)

Demodex folliculorum and D. brevis are cosmopolitan, obligatory parasites. Epidemiological studies have established a clear association between these species and various facial diseases in humans. However, not much is known of the ecology of these mites, also because it has proven difficult to culture them. Here we report a laboratory study on the (combined) effects of temperature and culture medium on the viability of both species. Results showed significant differences in the survival time of D. folliculorum and D. brevis among different temperatures and media. Pair-wise comparisons revealed that both species survived longest in serum medium, at a temperature of 16–22°C. Therefore, we conclude that these conditions allow preservation and maintenance of both Demodex mites.     

Phoresy of Trichouropoda shcherbakae Hirschmann, 1972 (Acari: Mesostigmata) on beetles of the genus Tetropium Kirby, 1837 (Coleoptera: Cerambycidae) in Białowieża Forest, Poland

The aim of this study was to assess the phoretic relationship between two beetle species of the genus Tetropium and mites from order Mesostigmata. The study was conducted in the Bia?owie?a Forest, which is recognized as one of Europe’s last natural forest areas. Insects were caught over a period of 8 weeks in 72 attractant traps (type Intercept IPM). In total 1250 specimens of genus Tetropium were collected. We analyzed 524 beetles, including 295 specimens of T. castaneum and 229 specimens of T. fuscum. On 49 beetles (9.4%) there were 785 individuals of Trichouropoda shcherbakae (Trematuridae). Mites were more common on T. fuscum, which carried 82% of all collected deutonymphs. Most of the mites found on beetles were attached to their legs. This study reports on changes in the intensity of phoresy in time and location of mite deutonymphs on their host species.     

Molecular evidence for host–parasite co-speciation between lizards and Schellackia parasites

Current and past parasite transmission may depend on the overlap of host distributions, potentially affecting parasite specificity and co-evolutionary processes. Nonetheless, parasite diversification may take place in sympatry when parasites are transmitted by vectors with low mobility. Here, we test the co-speciation hypothesis between lizard final hosts of the Family Lacertidae, and blood parasites of the genus Schellackia, which are potentially transmitted by haematophagous mites. The effects of current distributional overlap of host species on parasite specificity are also investigated. We sampled 27 localities on the Iberian Peninsula and three in northern Africa, and collected blood samples from 981 individual lizards of seven genera and 18 species. The overall prevalence of infection by parasites of the genus Schellackia was ～35%. We detected 16 Schellackia haplotypes of the 18S rRNA gene, revealing that the genus Schellackia is more diverse than previously thought. Phylogenetic analyses showed that Schellackia haplotypes grouped into two main monophyletic clades, the first including those detected in host species endemic to the Mediterranean region and the second those detected in host genera Acanthodactylus, Zootoca and Takydromus. All but one of the Schellackia haplotypes exhibited a high degree of host specificity at the generic level and 78.5% of them exclusively infected single host species. Some host species within the genera Podarcis (six species) and Iberolacerta (two species) were infected by three non-specific haplotypes of Schellackia, suggesting that host switching might have positively influenced past diversification of the genus. However, the results supported the idea that current host switching is rare because there existed a significant positive correlation between the number of exclusive parasite haplotypes and the number of host species with current sympatric distribution. This result, together with significant support for host–parasite molecular co-speciation, suggests that parasites of the genus Schellackia co-evolved with their lizard hosts.     

Biology and host specificity of the genusPsoroptes Gervais (Acarina: Psoroptidae), with reference to its occurrence in Australia

Psoroptes mites are obligate ectoparasites of mammals that cause psoroptic mange in a number of domestic and wildlife species world-wide. Correct identification of species in this genus is important for eradication and quarantine programs. However, conflicting reports about species identity and host specificity within this morphologically conservative genus have raised questions regarding the usefulness of the taxonomic characters previously used to distinguish species. Records that rely only on host species do not provide accurate identification ofPsoroptes mites at the species level. This paper reviews the information available on life history, geographic distribution, and host specificity for this genus, with special reference to problems associated with identifying species in Australia.     

How Diverse Is the Genus Wolbachia? Multiple-Gene Sequencing Reveals a Putatively New Wolbachia Supergroup Recovered from Spider Mites (Acari: Tetranychidae)

At least 20% of all arthropods and some nematode species are infected with intracellular bacteria of the genus Wolbachia. This highly diverse genus has been subdivided into eight “supergroups” (A to H) on the basis of nucleotide sequence data. Here, we report the discovery of a new Wolbachia supergroup recovered from the spider mite species Bryobia species V (Acari: Tetranychidae), based on the sequences of three protein-coding genes (ftsZ, gltA, and groEL) and the 16S rRNA gene. Other tetranychid mites possess supergroup B Wolbachia strains. The discovery of another Wolbachia supergroup expands the known diversity of Wolbachia and emphasizes the high variability of the genus. Our data also clarify the existing supergroup structure and highlight the use of multiple gene sequences for robust phylogenetic analysis. In addition to previous reports of recombination between the arthropod-infecting supergroups A and B, we provide evidence for recombination between the nematode-infecting supergroups C and D. Robust delineation of supergroups is essential for understanding the origin and spread of this common reproductive parasite and for unraveling mechanisms of host adaptation and manipulation across a wide range of hosts.     

Interspecific transmission of endosymbiotic Spiroplasma by mites

The occurrence of closely related strains of maternally transmitted endosymbionts in distantly related insect species indicates that these infections can colonize new host species by lateral transfer, although the mechanisms by which this occurs are unknown. We investigated whether ectoparasitic mites, which feed on insect haemolymph, can serve as interspecific vectors of Spiroplasma poulsonii, a male-killing endosymbiont of Drosophila. Using Spiroplasma-specific primers for PCR, we found that mites can pick up Spiroplasma from infected Drosophila nebulosa females and subsequently transfer the infection to Drosophila willistoni. Some of the progeny of the recipient D. willistoni were infected, indicating successful maternal transmission of the Spiroplasma within the new host species. However, the transmission rate of the infection from recipient flies to their offspring was low, perhaps due to low Spiroplasma density in the recipient flies.     

A new species of Leptobacillim,L. symbioticum,isolated from mites and sori of soybean rust

A verticillium-like fungus forming a whitish colony and mainly solitary phialides that produced fusiform to cylindrical conidia in chains was isolated from uredinia of Phakopsora pachyrhizi, a causal agent of soybean rust. In addition, two similar looking isolates were obtained from Prostigmata mites. Our taxonomic study based on morphology and phylogenetic analysis using ITS rDNA and LSU rDNA D1/D2 region sequences revealed that the three isolates were the same species and assigned to the genus Leptobacillium. These isolates represent a new species, morphologically and phylogenetically distinguished from the type species, L. leptobactrum, and we propose L. symbioticum sp. nov. In addition, Simplicillium chinense and S. coffeanum are assigned to the genus Leptobacillium.     

Host specificity in spinturnicid mites: do parasites share a long evolutionary history with their host?

Host specificity in parasites can be explained by spatial isolation from other potential hosts or by specialization and speciation of specific parasite species. The first assertion is based on allopatric speciation, the latter on differential lifetime reproductive success on different available hosts. We investigated the host specificity and cophylogenetic histories of four sympatric European bat species of the genus Myotis and their ectoparasitic wing mites of the genus Spinturnix. We sampled >40 parasite specimens from each bat species and reconstructed their phylogenetic COI trees to assess host specificity. To test for cospeciation, we compared host and parasite trees for congruencies in tree topologies. Corresponding divergence events in host and parasite trees were dated using the molecular clock approach. We found two species of wing mites to be host specific and one species to occur on two unrelated hosts. Host specificity cannot be explained by isolation of host species, because we found individual parasites on other species than their native hosts. Furthermore, we found no evidence for cospeciation, but for one host switch and one sorting event. Host‐specific wing mites were several million years younger than their hosts. Speciation of hosts did not cause speciation in their respective parasites, but we found that diversification of recent host lineages coincided with a lineage split in some parasites.     

Secondary structure prediction for complete rDNA sequences (18S, 5.8S,and 28S rDNA) of Demodex folliculorum, and comparison of divergent domains structures across Acari

According to base pairing, the rRNA folds into corresponding secondary structures, which contain additional phylogenetic information. On the basis of sequencing for complete rDNA sequences (18S, ITS1, 5.8S, ITS2 and 28S rDNA) of Demodex, we predicted the secondary structure of the complete rDNA sequence (18S, 5.8S, and 28S rDNA) of Demodex folliculorum, which was in concordance with that of the main arthropod lineages in past studies. And together with the sequence data from GenBank, we also predicted the secondary structures of divergent domains in SSU rRNA of 51 species and in LSU rRNA of 43 species from four superfamilies in Acari (Cheyletoidea, Tetranychoidea, Analgoidea and Ixodoidea). The multiple alignment among the four superfamilies in Acari showed that, insertions from Tetranychoidea SSU rRNA formed two newly proposed helixes, and helix c3-2b of LSU rRNA was absent in Demodex (Cheyletoidea) taxa. Generally speaking, LSU rRNA presented more remarkable differences than SSU rRNA did, mainly in D2, D3, D5, D7a, D7b, D8 and D10.     

A new species description of the acanthomus species group (Acari: Podapolipidae: Eutarsopolipus), with keys to world species of the group

Mites of the family Podapolipidae (Acari: Heterostigmatina) are permanent parasites of insects. During an intensive survey of insect parasitic mites in Iran, a new species of the genus Eutarsopolipus Berlese, 1911 (Acari: Heterostigmatina: Podapolipidae), belonging to the acanthomus species group, was found from the subelytra of Amara (Amara) aenea (De Geer, 1774) (Coleoptera: Carabidae). In this article, Eutarsopolipus amaraceus Tajodin and Hajiqanbar sp. nov. is described and compared with the most closely related species. In addition, keys to world species of all stages of the acanthomus group are provided, and the host range of all representatives of the group is discussed.     

Eriophyoid mites (Acari,Eriophyoidea) associated with tea plants,with descriptions of a new genus and two new species

A new genus and two new species of mites in the family Eriophyidae, Theaphyes rapaneae gen. n. and sp. n. which is found on the type host Rapanea neriifolia (Sieb. et Zucc.) Mez (Myrsinaceae) and Paracaphyllisa theacea sp. n., are described and illustrated. They are vagrants on the tea plant Camellia sinensis (L.) Kuntze and no apparent symptoms were detected. A key to the eriophyoid mites including thirteen species associated with tea plants all over the world is provided.     

Causal mechanisms underlying host specificity in bat ectoparasites

In parasites, host specificity may result either from restricted dispersal capacity or from fixed coevolutionary host-parasite adaptations. Knowledge of those proximal mechanisms leading to particular host specificity is fundamental to understand host-parasite interactions and potential coevolution of parasites and hosts. The relative importance of these two mechanisms was quantified through infection and cross-infection experiments using mites and bats as a model. Monospecific pools of parasitic mites (Spinturnix myoti and S. andegavinus) were subjected either to individual bats belonging to their traditional, native bat host species, or to another substitute host species within the same bat genus (Myotis). The two parasite species reacted differently to these treatments. S. myoti exhibited a clear preference for, and had a higher fitness on, its native host, Myotis myotis. In contrast, S. andegavinus showed no host choice, although its fitness was higher on its native host M. daubentoni. The causal mechanisms mediating host specificity can apparently differ within closely related host-parasite systems.