High temperatures and low humidity promote the occurrence of microsporidians (Microsporidia) in mosquitoes (Culicidae).

BACKGROUND: In the context of climate change, a growing concern is that vector-pathogen or host-parasite interactions may be correlated with climatic factors, especially increasing temperatures. In the present study, we used a mosquito-microsporidian model to determine the impact of environmental factors such as temperature, humidity, wind and rainfall on the occurrence rates of opportunistic obligate microparasites (Microsporidia) in hosts from a family that includes important disease vectors (Culicidae). METHODS: In our study, 3000 adult mosquitoes collected from the field over 3 years were analysed. Mosquitoes and microsporidia were identified using PCR and sequencing of the hypervariable V5 region of the small subunit ribosomal RNA gene and a shortened fragment of the cytochrome c oxidase subunit I gene, respectively. RESULTS: DNA metabarcoding was used to identify nine mosquito species, all of which were hosts of 12 microsporidian species. The prevalence of microsporidian DNA across all mosquito samples was 34.6%. Microsporidian prevalence in mosquitoes was more frequent during warm months (> 19 °C; humidity < 65%), as was the co-occurrence of two or three microsporidian species in a single host individual. During warm months, microsporidian occurrence was noted 1.6-fold more often than during the cold periods. Among the microsporidians found in the mosquitoes, five (representing the genera Enterocytospora, Vairimorpha and Microsporidium) were positively correlated with an increase in temperature, whereas one (Hazardia sp.) was significantly correlated with a decrease in temperature. Threefold more microsporidian co-occurrences were recorded in the warm months than in the cold months. CONCLUSIONS: These results suggest that the susceptibility of mosquitoes to parasite occurrence is primarily determined by environmental conditions, such as, for example, temperatures > 19 °C and humidity not exceeding 62%. Collectively, our data provide a better understanding of the effects of the environment on microsporidian-mosquito interactions.

Co-Occurrence of Borrelia burgdorferi Sensu Lato and Babesia spp. DNA in Ixodes ricinus Ticks Collected from Vegetation and Pets in the City of Poznan, Poland.

Here, we described the prevalence of Borrelia burgdorferi s.l. and Babesia species found in mono- and double infections among Ixodes ricinus ticks occurring in urban areas of the city of Poznan, Poland. We tested 1029 host-seeking ticks and 1268 engorged ticks removed from pet animals. Borrelia afzelii and B. garinii prevailed both in ticks from vegetation (3.7% and 3.7%, respectively) and from pets (3.7% and 0.6%, respectively). Babesia canis and Ba. microti were the most prevalent in host-seeking (2.6% and 1.4%, respectively) and feeding ticks (2.8% and 2.2%, respectively). Babesia microti sequences proved to be identical to the human pathogenic Ba. microti genotype "Jena/Germany". Sequences of the rarest piroplasm Ba. venatorum (0.7%) were identical with those isolated from European patients. About 1.0% of tested ticks yielded dual infections; in host-seeking ticks, Ba. canis prevailed in co-infections with B. afzelii and B. garinii, whereas Ba. microti and B. afzelii dominated in double-infected feeding ticks. Dual infections, even with a low prevalence, pose a challenge for differential diagnosis in patients with acute febrile disease after a tick bite. The finding of Ba. canis in both tick groups suggests that I. ricinus could be involved in the circulation of this piroplasm.

Evaluation of Genetic Diversity in Quill Mites of the Genus Syringophiloidus Kethley, 1970 (Prostigmata: Syringophilidae) with Six New-to-Science Species.

Quill mites (Acariformes: Syringophilidae) are poorly explored bird parasites. Syringophiloidus Kethley, 1970, is the most specious and widespread genus in this family. It is believed to contain mono-, steno- and poly-xenous parasites and thus seems to be an exemplary for studies on biodiversity and host associations. In this work, we applied the DNA barcode marker (mitochondrial cytochrome c oxidase subunit I gene fragment, COI) to analyze the species composition and host specificity of representatives of fifteen Syringophiloidus populations parasitizing fifteen bird species. The neighbor joining analyses distinguished thirteen monophyletic lineages, almost completely corresponding to seven previously known species recognized based on morphological features, and six new-to-science species. The only exception is S. amazilia Skoracki, 2017, which is most likely conspecific with Syringophiloidus stawarczyki Skoracki, 2004. The intraspecific distances of all species were not higher than 0.9%, whilst the interspecific diversity ranged from 5.9% to 19.2% and 6.3-22.4%, inferred as the distances p and K2P, respectively. Although all putative species (except S. amazilia) are highly supported, the relationships between them have not been fully resolved and only faintly indicate that both host phylogeny and distributions influence the phylogenetic structure of quill mite taxa.

Differences in the proliferation trend of 'Microsporidium' sp. PL03 in Culex pipiens and C. torrentium larvae.

Our study aimed to examine whether there are differences in the proliferation trend of microsporidia in mosquito larvae of the same genus (Culex spp.). DNA-barcoding and quantitative analyses were used to determine microsporidian rDNA copies in 'early' (L1 + L2) and 'late' (L3 + L4) Culex larvae in a natural population. In the study area, C. pipiens and C. torrentium larvae were infected by 'Microsporidium' sp. PL03 at similar levels. Infection by this microsporidian species probably elicits a notable immune response in C. pipiens, whereas in C. torrentium, it may evade or suppress the host immune response.

Patrilocality and hunter-gatherer-related ancestry of populations in East-Central Europe during the Middle Bronze Age.

The demographic history of East-Central Europe after the Neolithic period remains poorly explored, despite this region being on the confluence of various ecological zones and cultural entities. Here, the descendants of societies associated with steppe pastoralists form Early Bronze Age were followed by Middle Bronze Age populations displaying unique characteristics. Particularly, the predominance of collective burials, the scale of which, was previously seen only in the Neolithic. The extent to which this re-emergence of older traditions is a result of genetic shift or social changes in the MBA is a subject of debate. Here by analysing 91 newly generated genomes from Bronze Age individuals from present Poland and Ukraine, we discovered that Middle Bronze Age populations were formed by an additional admixture event involving a population with relatively high proportions of genetic component associated with European hunter-gatherers and that their social structure was based on, primarily patrilocal, multigenerational kin-groups.

Exposure of dogs and cats to Borrelia miyamotoi infected Ixodes ricinus ticks in urban areas of the city of Poznan, west-central Poland.

Borrelia miyamotoi is an emerging human pathogen that causes a relapsing fever-like disease named B. miyamotoi disease. The bacterium belongs to the relapsing fever borreliae, and similar to spirochetes of the Borrelia burgdorferi sensu lato group, it is transmitted only by hard ticks of the Ixodes ricinus complex. To date, B. miyamotoi has not been demonstrated to cause illness in dogs or cats, and is poorly documented in veterinary medicine. The aim of this study was to determine the B. miyamotoi presence in (i) host-seeking ticks and (ii) engorged Ixodes sp. ticks collected from dogs and cats during their inspection in veterinary clinics of the city of Poznan, west-central Poland. Host-seeking ticks were sampled in dog walking areas localized in urban forested recreational sites of the city. In this study, 1,059 host-seeking and 837 engorged I. ricinus ticks collected from 680 tick-infested animals (567 dogs and 113 cats) were screened. Additionally, 31 I. hexagonus ticks (one larva, 13 nymphs, and 17 females) were collected from three cats; one larva and one nymph were collected from two dogs; and one dog was infested with a single Dermacentor reticulatus female. Borrelia DNA was identified by the amplification and sequencing of the V4 hypervariable region of the 16S rRNA gene and flaB gene fragments. DNA of B. miyamotoi was detected in 22 (2.1%) of the host-seeking ticks (in all developmental tick stages and in all study areas). In addition, the engorged I. ricinus ticks exhibited a similar B. miyamotoi presence (1.8%). Fifteen I. ricinus ticks collected from animals tested positive for the presence of B. miyamotoi DNA, and the DNA of B. miyamotoi was observed in three (9.1%; one female and two nymphs) I. hexagonus ticks. The single D. reticulatus female collected from a dog tested PCR-negative for the bacterium. The results of this study demonstrated the establishment and broad presence of the bacterium in tick populations from different urban ecosystems of the city of Poznan. The lack of difference in the mean infection presence of animal-derived and host-seeking I. ricinus ticks suggests that the systematic surveillance of pets may be useful for the evaluation of human exposure to B. miyamotoi infected ticks in urban areas. Additional studies are required to further elucidate the role of domestic and wild carnivores in the epidemiology of B. miyamotoi, which remains unknown.

Glyphosate versus glyphosate based ionic liquids: Effect of cation on glyphosate biodegradation, soxA and phnJ genes abundance and microbial populations changes during soil bioaugmentation.

The applicability of herbicidal ionic liquids (HILs) as an alternative form of herbicides is currently evaluated. Yet, the available research is lacking information on the behaviour of herbicidal ionic liquids upon addition to the environment, i.e., if cations and anions act as separate moieties or remain an ionic salt. Hence, we tested degradation of five HILs with the glyphosate anion, their bioavailability in soil, toxicity towards microorganisms, impact on the biodiversity and the abundance of phnJ and soxA genes. The cations were proven to be slightly or moderately toxic. The properties of cations determined the properties of the whole formulation, which might suggest that cations and anion act as the independent mixture of ions. The mineralisation efficiencies were in the range of 15-53%; however, in the case of cations (except non-toxic choline), only 13-20% were bioavailable for degradation. The hydrophobic cations were proven to be highly sorbed, while the anion was readily available for microbial degradation regardless of its counterion. The approach to enrich test samples with isolated microorganisms specialised in glyphosate degradation resulted in higher degradation efficiencies, yet not high enough to mitigate the negative impact of cations. In addition, increased activity of enzymes participating in glyphosate degradation was observed. In the view of obtained results, the use of cationic surfactants in HILs structure is not recommended, as sorption was shown to be determining factor in HILs degradation efficiency. Moreover, obtained results indicate that corresponding ions in HILs might act as separate moieties in the environment.

Microsporidian Infection in Mosquitoes (Culicidae) Is Associated with Gut Microbiome Composition and Predicted Gut Microbiome Functional Content.

The animal gut microbiota consist of many different microorganisms, mainly bacteria, but archaea, fungi, protozoans, and viruses may also be present. This complex and dynamic community of microorganisms may change during parasitic infection. In the present study, we investigated the effect of the presence of microsporidians on the composition of the mosquito gut microbiota and linked some microbiome taxa and functionalities to infections caused by these parasites. We characterised bacterial communities of 188 mosquito females, of which 108 were positive for microsporidian DNA. To assess how bacterial communities change during microsporidian infection, microbiome structures were identified using 16S rRNA microbial profiling. In total, we identified 46 families and four higher taxa, of which Comamonadaceae, Enterobacteriaceae, Flavobacteriaceae and Pseudomonadaceae were the most abundant mosquito-associated bacterial families. Our data suggest that the mosquito gut microbial composition varies among host species. In addition, we found a correlation between the microbiome composition and the presence of microsporidians. The prediction of metagenome functional content from the 16S rRNA gene sequencing suggests that microsporidian infection is characterised by some bacterial species capable of specific metabolic functions, especially the biosynthesis of ansamycins and vancomycin antibiotics and the pentose phosphate pathway. Moreover, we detected a positive correlation between the presence of microsporidian DNA and bacteria belonging to Spiroplasmataceae and Leuconostocaceae, each represented by a single species, Spiroplasma sp. PL03 and Weissella cf. viridescens, respectively. Additionally, W. cf. viridescens was observed only in microsporidian-infected mosquitoes. More extensive research, including intensive and varied host sampling, as well as determination of metabolic activities based on quantitative methods, should be carried out to confirm our results.

Microsporidians (Microsporidia) parasitic on mosquitoes (Culicidae) in central Europe are often multi-host species.

Microsporidians (Microsporidia) are a diverse group of obligate and intracellular parasites of eukaryotes. There is evidence that the real species diversity in the phylum could be greatly underestimated, especially for microsporidians parasitic on invertebrates. Mosquitoes (Culicidae) are among very important microsporidian host groups. However, to date, no extensive survey on the prevalence of microsporidians in European mosquitoes has been performed. Here, we used mosquitoes collected in west-central Poland and a metabarcoding approach to examine the prevalence and diversity of microsporidian species among European mosquitoes. We found that up to one-third of mosquitoes in Europe may be infected with at least 13 microsporidian species belonging to the genera Amblyospora, Hazardia, Encephalitozoon, Enterocytospora, and Nosema and the holding genus Microsporidium. The lack of a difference in microsporidian prevalence between mosquito sexes implies that other factors, e.g., temperature or humidity, affect microsporidian occurrence in adult mosquitoes. Each microsporidian species was found in at least three mosquito species, which suggests that these microsporidians are polyxenic rather than monoxenic parasites. The co-occurrence of at least two different microsporidian species was found in 3.6% of host individuals. The abundance of microsporidian DNA sequences suggests interactions between co-occurring parasites; however, these results should be confirmed by microscopic and quantitative methods. In addition, further histological research is required to describe Microsporidium sp. PL01 or match its DNA to that of an already described species.

Trophic and symbiotic links between obligate-glacier water bears (Tardigrada) and cryoconite microorganisms.

Insights into biodiversity and trophic webs are important for understanding ecosystem functions. Although the surfaces of glaciers are one of the most productive and biologically diverse parts of the cryosphere, the links between top consumers, their diet and microbial communities are poorly understood. In this study, for the first time we investigated the relationships between bacteria, fungi and other microeukaryotes as they relate to tardigrades, microscopic metazoans that are top consumers in cryoconite, a biologically rich and productive biogenic sediment found on glacier surfaces. Using metabarcoding (16S rDNA for bacteria, ITS1 for fungi, and 18S rDNA for other microeukaryotes), we analyzed the microbial community structures of cryoconite and compared them with the community found in both fully fed and starved tardigrades. The community structure of each microbial group (bacteria, fungi, microeukaryotes) were similar within each host group (cryoconite, fully fed tardigrades and starved tardigrades), and differed significantly between groups, as indicated by redundancy analyses. The relative number of operational taxonomic units (ZOTUs, OTUs) and the Shannon index differed significantly between cryoconite and tardigrades. Species indicator analysis highlighted a group of microbial taxa typical of both fully fed and starved tardigrades (potential commensals), like the bacteria of the genera Staphylococcus and Stenotrophomonas, as well as a group of taxa typical of both cryoconite and fully fed tardigrades (likely part of the tardigrade diet; bacteria Flavobacterium sp., fungi Preussia sp., algae Trebouxiophyceae sp.). Tardigrades are consumers of bacteria, fungi and other microeukaryotes in cryoconite and, being hosts for diverse microbes, their presence can enrich the microbiome of glaciers.

Metabarcoding reveals low prevalence of microsporidian infections in castor bean tick (Ixodes ricinus).

BACKGROUND: Microsporidia is a large group of eukaryotic obligate intracellular spore-forming parasites, of which 17 species can cause microsporidiosis in humans. Most human-infecting microsporidians belong to the genera Enterocytozoon and Encephalitozoon. To date, only five microsporidian species, including Encephalitozoon-like, have been found in hard ticks (Ixodidae) using microscopic methods, but no sequence data are available for them. Furthermore, no widespread screening for microsporidian-infected ticks based on DNA analysis has been carried out to date. Thus, in this study, we applied a recently developed DNA metabarcoding method for efficient microsporidian DNA identification to assess the role of ticks as potential vectors of microsporidian species causing diseases in humans. METHODS: In total, 1070 (493 juvenile and 577 adult) unfed host-seeking Ixodes ricinus ticks collected at urban parks in the city of Poznan, Poland, and 94 engorged tick females fed on dogs and cats were screened for microsporidian DNA. Microsporidians were detected by PCR amplification and sequencing of the hypervariable V5 region of 18S rRNA gene (18S profiling) using the microsporidian-specific primer set. Tick species were identified morphologically and confirmed by amplification and sequencing of the shortened fragment of cytochrome c oxidase subunit I gene (mini-COI). RESULTS: All collected ticks were unambiguously assigned to I. ricinus. Potentially zoonotic Encephalitozoon intestinalis was identified in three fed ticks (3.2%) collected from three different dogs. In eight unfed host-seeking ticks (0.8%), including three males (1.1%), two females (0.7%) and three nymphs (0.7%), the new microsporidian sequence representing a species belonging to the genus Endoreticulatus was identified. CONCLUSIONS: The lack of zoonotic microsporidians in host-seeking ticks suggests that I. ricinus is not involved in transmission of human-infecting microsporidians. Moreover, a very low occurrence of the other microsporidian species in both fed and host-seeking ticks implies that mechanisms exist to defend ticks against infection with these parasites.

Prevalence of Babesia canis DNA in Ixodes ricinus ticks collected in forest and urban ecosystems in west-central Poland.

Babesia canis, a widely distributed European tick-borne protozoan haemoparasite, causes canine babesiosis, the most important tick-borne disease afflicting dogs worldwide. The meadow tick, Dermacentor reticulatus, is considered to be the primary vector of this parasite in central Europe. Females of the more broadly distributed and medically important castor bean tick, Ixodes ricinus, also commonly feed upon dogs, but their role in the enzootic transmission cycle of B. canis is unclear. Here, we screened 1,598 host-seeking I. ricinus ticks collected from two different ecosystems, forest stands vs. urban recreational forests, for the presence of B. canis DNA. Ticks were sampled during their two seasonal peaks of activity, spring (May/June) and late summer (September). Babesia species were identified by amplification and sequencing of a hypervariable 18S rRNA gene fragment. Babesia canis was the only piroplasm detected in 13% of 200 larvae and 8.2% of 324 nymphs in the forest ecosystems. In urban recreational areas, B. canis DNA was found in 1.5% of 460 nymphs, 3.5% of 289 females and 3.2% of 280 males. Additionally, three samples, including one female, one male, and one nymph, were co-infected with B. venatorum and one nymph with B. divergens or B. capreoli. Our findings implicate that B. canis can be transmitted transovarially and maintained transstadially within populations of I. ricinus, but the vector competence of I. ricinus for transmitting B. canis remains to be investigated.

Maternal genetic origin of the late and final Neolithic human populations from present-day Poland.

OBJECTIVE: We aim to identify maternal genetic affinities between the Middle to Final Neolithic (3850-2300 BC) populations from present-day Poland and possible genetic influences from the Pontic steppe. MATERIALS AND METHODS: We conducted ancient DNA studies from populations associated with Zlota, Globular Amphora, Funnel Beaker, and Corded Ware cultures (CWC). We sequenced genomic libraries on Illumina platform to generate 86 complete ancient mitochondrial genomes. Some of the samples were enriched for mitochondrial DNA using hybridization capture. RESULTS: The maternal genetic composition found in Zlota-associated individuals resembled that found in people associated with the Globular Amphora culture which indicates that both groups likely originated from the same maternal genetic background. Further, these two groups were closely related to the Funnel Beaker culture-associated population. None of these groups shared a close affinity to CWC-associated people. Haplogroup U4 was present only in the CWC group and absent in Zlota group, Globular Amphora, and Funnel Beaker cultures. DISCUSSION: The prevalence of mitochondrial haplogroups of Neolithic farmer origin identified in Early, Middle and Late Neolithic populations suggests a genetic continuity of these maternal lineages in the studied area. Although overlapping in time - and to some extent - in cultural expressions, none of the studied groups (Zlota, Globular Amphora, Funnel Beaker), shared a close genetic affinity to CWC-associated people, indicating a larger extent of cultural influence from the Pontic steppe than genetic exchange. The higher frequency of haplogroup U5b found in populations associated with Funnel Beaker, Globular Amphora, and Zlota cultures suggest a gradual maternal genetic influx from Mesolithic hunter-gatherers. Moreover, presence of haplogroup U4 in Corded Ware groups is most likely associated with the migrations from the Pontic steppe at the end of the Neolithic and supports the observed genetic distances.

Zoonotic Giardia duodenalis sub-assemblage BIV in wild raccoons (Procyon lotor) from Germany and Luxembourg.

Giardia duodenalis is a cosmopolitan flagellate that causes giardiasis, one of the most significant gastrointestinal diseases in humans. This parasite can be a serious threat to public health because it can cause waterborne outbreaks as well as sporadic infections in humans. Invasive raccoons (Procyon lotor) may play a role in disseminating Giardia into the environment and transmitting it to humans and domestic animals because they live in high densities and deposit their faces in latrines near areas used by humans. While Giardia infections have been reported from raccoons in North America, it is unknown whether they carry G. duodenalis with zoonotic assemblage A and B, which have the potential to cause illness in humans. We collected faecal samples from 66 legally harvested raccoons in Germany and Luxembourg and examined for Giardia using molecular techniques. Using a quantitative PCR based on primers specific to Giardia genetic assemblages A and B, we detected the presence of zoonotic assemblage B in 27% (95% CI, 17.0-39.6) of all examined faecal samples from raccoons, including animals sampled in buildings. We did not detect genetic assemblage A in any of the samples. Sequences obtained from the glutamate dehydrogenase and beta-giardin gene fragments from a selection of three of the positive samples showed that raccoons carried a zoonotic G. duodenalis genotype belonging to sub-assemblage BIV, which is commonly found in humans and animals worldwide. Our results suggest that free-ranging raccoons have the potential to play an increasingly important role in the epidemiology of Giardia and pose a threat to public health in Europe and other regions where this species is common and lives in close association with humans.

Correction:Analysis of LCT-13910 genotypes and bone mineral density in ancient skeletal materials.

[This corrects the article DOI: 10.1371/journal.pone.0194966.].

A new method of metabarcoding Microsporidia and their hosts reveals high levels of microsporidian infections in mosquitoes (Culicidae).

DNA metabarcoding offers new perspectives, especially with regard to the high-throughput identification and diagnostics of pathogens. Microsporidia are an example of widely distributed, opportunistic and pathogenic microorganisms in which molecular identification is important for both environmental research and clinical diagnostics. We have developed a method for parallel detection of both microsporidian infection and the host species. We designed new primer sets: one specific for the classical Microsporidia (targeting the hypervariable V5 region of small subunit [ssu] rDNA), and a second one targeting a shortened fragment of the COI gene (standard metazoan DNA-barcode); both markers are well suited for next generation sequencing. Analysis of the ssu rDNA data set representing 607 microsporidian species (120 genera) indicated that the V5 region enables identification of >98% species in the data set (596/607). To test the method, we used microsporidians that infect mosquitoes in natural populations. Using mini-COI data, all field-collected mosquitoes were unambiguously assigned to seven species; among them almost 60% of specimens were positive for at least 11 different microsporidian species, including a new microsporidian ssu rDNA sequence (Microsporidium sp. PL01). Phylogenetic analysis showed that this species belongs to one of the two main clades in the Terresporidia. We found a high rate of microsporidian co-infections (9.4%). The numbers of sequence reads for the operational taxonomic units suggest that the occurrence of Nosema spp. in co-infections could benefit them; however, this observation should be retested using a more intensive host sampling. Our results show that DNA barcoding is a rapid and cost-effective method for deciphering sample diversity in greater resolution, including the hidden biodiversity that may be overlooked using classical methodology.

Mitochondrial genomes from Bronze Age Poland reveal genetic continuity from the Late Neolithic and additional genetic affinities with the steppe populations.

OBJECTIVE: In this work we aim to investigate the origins and genetic affinities of Bronze Age populations (2,400-1,100 BC) from the region of southern Poland and to trace maternal kinship patterns present in the burials of those populations by the use of complete mitochondrial genomes. MATERIALS AND METHODS: We performed ancient DNA analyses for Bronze Age individuals from present-day Poland associated with the Strzyzow culture, the Mierzanowice culture, and the Trzciniec Cultural circle. To obtain complete mitochondrial genomes, we sequenced genomic libraries using Illumina platform. Additionally, hybridization capture was used to enrich some of the samples for mitochondrial DNA. AMS 14 C-dating was conducted for 51 individuals to verify chronological and cultural attribution of the analyzed samples. RESULTS: Complete ancient mitochondrial genomes were generated for 80 of the Bronze Age individuals from present-day Poland. The results of the population genetic analyses indicate close maternal genetic affinity between Mierzanowice, Trzciniec, and Corded Ware culture-associated populations. This is in contrast to the genetically more distant Strzyzów people that displayed closer maternal genetic relation to steppe populations associated with the preceding Yamnaya culture and Catacomb culture, and with later Scythians. Potential maternal kinship relations were identified in burials of Mierzanowice and Trzciniec populations analyzed in this study. DISCUSSION: Results revealed genetic continuity from the Late Neolithic Corded Ware groups to Bronze Age Mierzanowice and Trzciniec-associated populations, and possible additional genetic contribution from the steppe to the formation of the Strzyzów-associated group at the end of 3rd millennium BC. Mitochondrial patterns indicated several pairs of potentially maternally related individuals mostly in Trzciniec-associated group.

Microbial composition of enigmatic bird parasites: Wolbachia and Spiroplasma are the most important bacterial associates of quill mites (Acariformes: Syringophilidae).

BACKGROUND: The microbiome is an integral component of many animal species, potentially affecting behavior, physiology, and other biological properties. Despite this importance, bacterial communities remain vastly understudied in many groups of invertebrates, including mites. Quill mites (Acariformes: Syringophilidae) are a poorly known group of permanent bird ectoparasites that occupy quills of feathers and feed on bird subcutaneous tissue and fluids. Most of the known species have strongly female-biased sex ratio, and it was hypothesized that this is caused by endosymbiotic bacteria. Previously, Anaplasma phagocytophilum (Foggie) and a high diversity of Wolbachia strains were detected in quill mites via targeted PCR screens. Here, we use an unbiased 16S rRNA gene amplicon sequencing approach to determine other bacteria that potentially impact quill mite biology. RESULTS: We performed 16S rRNA gene amplicon sequencing of 126 quill mite individuals from eleven species parasitizing twelve species (four families) of passeriform birds. In addition to Wolbachia, we found Spiroplasma as potential symbiont of quill mites. Consistently, high Spiroplasma titers were only found in individuals of two mite species associated with finches of the genus Carduelis, suggesting a history of horizontal transfers of Spiroplasma via the bird host. Furthermore, there was evidence for Spiroplasma negatively affecting Wolbachia titers. We found no evidence for the previously reported Anaplasma in quill mites, but detected sequences of high similarity to the potential pathogens Brucella and Bartonella at low abundances. Other amplicon sequence variants (ASVs) could be assigned to a diverse number of bacterial taxa, including several that were previously isolated from bird skin. Further, many frequently found ASVs were assigned to taxa that show a very broad distribution with no strong prior evidence for symbiotic association with animals. We interpret these findings as evidence for a scarcity of resident microbial associates (other than inherited symbionts) in quill mites.

Molecular phylogeny of Eupodidae reveals that the family Cocceupodidae (Actinotrichida; Eupodoidea) and its genus Filieupodes are valid taxa.

The family Eupodidae is one of eight families belonging to the superfamily Eupodoidea. Although the Eupodidae are very common mites, they are still poorly investigated and the systematic parameters of the family are vague with unclear intrafamilial relationships between its genera. According to the hypothesis of Jesionowska (Genus 21(4):637-658, 2010), the eupodid genera Cocceupodes, Linopodes and her new genus Filieupodes should be transferred from the Eupodidae to a separate family named Cocceupodidae. The opposite hypothesis of Khaustov (ZooKeys 422:11-22, 2014) considers the family Cocceupodidae an artificial taxon and the genus Filieupodes a junior synonym of Cocceupodes and assignes the family Eupodidae its primary definition. Here, we present the first molecular reconstruction of phylogenetic relationships within the Eupodidae. We test the taxonomic status of the Eupodidae and refer to the previous hypotheses based on morphological data. According to the reconstructed phylogram based on nuclear (18S + 28S rDNA) and mitochondrial (COI) sequences as well as morphological characters tracing this analysis, we have concluded that: (1) the genera Linopodes, Filieupodes and Cocceupodes are a separate monophyletic lineage of familial rank and the sister group of Eupodidae s.s., and (2) the genus Filieupodes should be considered a separate genus being a sister group of the Linopodes-Cocceupodes clade.