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.

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.

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.

Sweet or salty? The origin of freshwater gastrotrichs (Gastrotricha, Chaetonotida) revealed by molecular phylogenetic analysis.

Chaetonotidae is the most diverse and widely distributed family of the order Chaetonotida (Gastrotricha) and includes both marine and freshwater species. Although the family is regarded as a sister taxon to the exclusively marine Xenotrichulidae, the type of environment, marine or freshwater, where Chaetonotidae originated is still not known. Here, we reconstructed the phylogeny of the family based on molecular sequence data and mapped both morphological and ecological characters to determine the ancestral environment of the first members of the family. Our results revealed that the freshwater genus Bifidochaetus is the earliest branching lineage in the paraphyletic Chaetonotidae (encompassing Dasydytidae and Neogosseidae). Moreover, we reconstructed Lepidochaetus-Cephalionotus clade as a monophyletic sister group to the remaining chaetonotids, which supports Kisielewski's morphological based hypothesis concerning undifferentiated type of body scales as a most primary character in Chaetonotidae. We also found that reversals to marine habitats occurred independently in different Chaetonotidae lineages, thus marine species in the genera Heterolepidoderma, Halichaetonotus, Aspidiophorus and subgenera Chaetonotus (Schizochaetonotus) or Chaetonotus (Marinochaetus) should be assumed as having secondarily invaded the marine environment. Character mapping revealed a series of synapomorphies that define the clade that includes Chaetonotidae (with Dasydytidae and Neogosseidae), the most important of which may be those linked to reproduction.

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.

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.

The Reappearance of An Extremely Rare and Critically Endangered Nitella translucens (Charophyceae) in Poland.

We report the reappearance of the rare charophyte Nitella translucens in Poland. It was identified in the soft-water lobelian Lake Jelen (North Poland) during 2013 and 2018 phytolittoral surveys. This species is considered critically endangered in various European countries and was previously classified as extinct in Poland. Its occurrence was confirmed using morphological and molecular data (ITS1-18S, ITS2-28S, rDNA, and rbcL). The N. translucens occupied ~20% of the lake bottom, at depths of 1.5-6.5 m, water pH 7.5-8.6, conductivity of 59-66 µS · cm-1 , and total nitrogen and phosphorus during growing season in the range of 1.1-1.4 mg · L-1 and 0.07-0.1 mg · L-1 , respectively. It co-occurred mainly with plant species typical for lobelia lakes: Isoetes lacustris, Littorella uniflora, and Myriophyllum alterniflorum, as well as Ceratophyllum demersum and Elodea canadensis, which are characteristic for eutrophicated waters. It appears that N. translucens may thrive in lobelia lakes during their transformation to more eutrophic states.

Combined description (morphology with DNA barcode data) of a new quill mite Torotrogla paenae n. sp. (Acariformes: Syringophilidae) parasitising the Kalahari scrub-robin Cercotrichas paena (Smith) (Passeriformes: Muscicapidae) in Namibia.

A new quill mite species Torotrogla paenae n. sp. (Acariformes: Syringophilidae) parasitising the Kalahari scrub-robin Cercotrichas paena (Smith) (Passeriformes: Muscicapidae) in Namibia is described based on the external morphology and DNA barcode data (the mitochondrial cytochrome c oxidase subunit 1 sequences, cox1). Females of T. paenae n. sp. morphologically differ from the most similar species T. lusciniae Skoracki, 2004 by the total body length (780-830 vs 645-715 µm in T. lusciniae) and the presence of hysteronotal shields (vs absence), apunctate propodonotal and pygidial shields (vs punctate), apunctate coxal fields (vs punctate), the fan-like setae p' and p" of legs III-IV provided with c.10 tines (vs 14-15) and the length of setae si (140-180 vs 190-210 µm) and se (160-185 vs 210-225 µm). The male of T. paenae n. sp. morphologically differs from T. lusciniae by the lateral branch of peritremes composed of 4 chambers (vs 7-8 chambers) and lengths of setae ve (45 vs 70-75 µm) and se (120 vs 165 µm).

Late Danubian mitochondrial genomes shed light into the Neolithisation of Central Europe in the 5th millennium BC.

BACKGROUND: Recent aDNA studies are progressively focusing on various Neolithic and Hunter - Gatherer (HG) populations, providing arguments in favor of major migrations accompanying European Neolithisation. The major focus was so far on the Linear Pottery Culture (LBK), which introduced the Neolithic way of life in Central Europe in the second half of 6th millennium BC. It is widely agreed that people of this culture were genetically different from local HGs and no genetic exchange is seen between the two groups. From the other hand some degree of resurgence of HGs genetic component is seen in late Neolithic groups belonging to the complex of the Funnel Beaker Cultures (TRB). Less attention is brought to various middle Neolithic cultures belonging to Late Danubian sequence which chronologically fall in between those two abovementioned groups. We suspected that genetic influx from HG to farming communities might have happened in Late Danubian cultures since archaeologists see extensive contacts between those two communities. RESULTS: Here we address this issue by presenting 5 complete mitochondrial genomes of various late Danubian individuals from modern-day Poland and combining it with available published data. Our data show that Late Danubian cultures are maternally closely related to Funnel Beaker groups instead of culturally similar LBK. CONCLUSIONS: We assume that it is an effect of the presence of individuals belonging to U5 haplogroup both in Late Danubians and the TRB. The U5 haplogroup is thought to be a typical for HGs of Europe and therefore we argue that it is an additional evidence of genetic exchange between farming and HG groups taking place at least as far back as in middle Neolithic, in the Late Danubian communities.

Higher-level molecular phylogeny of the water mites (Acariformes: Prostigmata: Parasitengonina: Hydrachnidiae).

With nearly 6000 named species, water mites (Hydrachnidiae) represent the largest group of arachnids to have invaded and extensively diversified in freshwater habitats. Water mites together with three other lineages (the terrestrial Erythraiae and Trombidiae, and aquatic Stygothrombiae), make up the hyporder Parasitengonina, which is characterized by having parasitic larvae and predatory nymphs and adults. Relationships between the Hydrachnidiae and other members of the Parasitengonina are unclear, as are relationships among the major lineages of water mites. Monophyly of water mites has been asserted, with the possible exception of the morphologically distinctive Hydrovolzioidea. Here we infer the phylogeny of water mites using multiple molecular markers and including representatives of all superfamilies of Hydrachnidiae and of almost all other Parasitengonina. Our results support a monophyletic Parasitengonina including Trombidiae, Stygothrombiae, and Hydrachnidiae. A monophyletic Hydrachnidiae, including Hydrovolzioidea, is strongly supported. Terrestrial Parasitengonina do not form a monophyletic sister group to water mites. Stygothrombiae is close to water mites but is not nested within this clade. Water mites appear to be derived from ancestors close to Stygothrombiae or the erythraoid group Calyptostomatoidea; however, this relationship is not clear because of extremely short branches in this part of the parasitengonine tree. We recovered with strong support all commonly accepted water mite superfamilies except for Hydryphantoidea, which is clearly paraphyletic. Our data support the previously proposed clades Protohydrachnidia (Hydrovolzioidea and Eylaoidea), Euhydrachnidia (all remaining superfamilies), and the euhydrachnid subclade Neohydrachnidia (Lebertioidea, Hydrachnoidea, Hygrobatoidea, and Arrenuroidea). We found that larval leg structure and locomotory behavior are strongly congruent with the molecular phylogeny. Other morphological and behavioral characters, including host choice, are not as strongly correlated with phylogeny. Molecular dating suggests that the Hydrachnidiae arose about 235MYA, and that Neohydrachnidia began to diversify about 155MYA. Our results provide a strong framework for classification and for further elaboration at finer taxonomic scales, which will allow testing of ecological and behavioral hypotheses associated with the transition from terrestrial to aquatic life.

Three new species of the genus Pterygosoma Peters, 1849 (Acariformes: Pterygosomatidae) from agamid lizards (Sauria: Agaminae) with DNA barcode data.

Three new species of the genus Pterygosoma Peters, 1849 parasitising lizards of the subfamily Agaminae (Squamata: Agamidae) are described: P. pallidum n. sp. from Trapelus pallidus (Merrem) and P. parasiniatum n. sp. from Pseudotrapelus cf. sinaitus (Heyden) (both from Jordan); and P. theobaldi n. sp. from Phrynocephalus theobaldi Blyth from North India. We extend the standard morphological description of the new species by using DNA barcode markers, partial sequences of the mitochondrial cytochrome c oxidase subunit I (cox1) gene and the hypervariable region D2 of the nuclear 28S rRNA gene. A key to the species group inermis is constructed based on female morphology. The agamid genus Phrynocephalus Kaup, 1825 is recorded as a host of Pterygosoma for the first time.

Aulonastus similis n. sp., a new quill mite species (Syringophilidae) parasitising passeriform birds (Tyrannidae and Cardinalidae) in Mexico.

A new quill mite species, Aulonastus similis n. sp. (Acariformes: Syringophilidae), parasitising Myiozetetes similis (Spix) (Tyrannidae) and Habia fuscicauda (Cabanis) (Cardinalidae) in Mexico is described and DNA barcode sequences of the mitochondrial cytochrome c oxidase subunit I (cox1) and D1-D3 region of the nuclear 28S rRNA gene are provided. Morphologically, females of A. similis are close to A. euphagus Skoracki, Hendricks & Spicer, 2010 but differ from this species in the length ratios of the idiosomal setae: ve:si (2-2.3:1 vs 1:1) and f2:f1 (4.7-6.3:1 vs 3.3:1).

Differences in speciation progress in feather mites (Analgoidea) inhabiting the same host: the case of Zachvatkinia and Alloptes living on arctic and long-tailed skuas.

Recent molecular phylogenetic analyses have revealed that some apparently oligoxenous feather mite species are in fact monoxenous cryptic species with little morphological differentiation. In this study we analyzed two species, Zachvatkinia isolata (Avenzoariidae) and Alloptes (Sternalloptes) stercorarii (Alloptidae) which prefer different parts of the plumage of two sister species of birds: arctic skua (Stercorarius parasiticus) and long-tailed skua (S. longicaudus) breeding on tundra in the High Arctic archipelago of Svalbard. Given that there are no reports about hybridization events between the host species, we expected that both skuas would have a species-specific acarofauna. The genetic distances among DNA-barcode sequences (COI and 28S rDNA), phylogenetic tree topologies, and haplotype networks of the COI sequences of mites suggested extensive gene flow in Z. isolata between and within populations inhabiting both skua species, whereas the Alloptes populations were host specific and sufficiently genetically separated as to warrant species-level status. The discrepancy in the genetic structure of Alloptes and Zachvatkinia populations suggests frequent but transient contacts between the two skua species in which the probability of mite exchange is much higher for Zachvatkinia, which is present in high numbers and inhabits exposed parts of primary flight feathers, than for the less abundant Alloptes that lives primarily in more protected and inaccessible parts of the plumage. We discuss the possible nature of these contacts between host species and the area(s) where they might take place. The star-like structures in the haplotype network as well as high haplotype diversity and low nucleotide diversity observed in Z. isolata are concordant with the known dispersal strategy of feather mites: vertical colonization of new host individuals followed by rapid growth of founder populations.

Phylogenetic position of the enigmatic clawless eutardigrade genus Apodibius Dastych, 1983 (Tardigrada), based on 18S and 28S rRNA sequence data from its type species A. confusus.

The systematics of Eutardigrada, the largest lineage among the three classes of the phylum Tardigrada, is based mainly on the morphology of the leg claws and of the buccal apparatus. However, three members of the rarely recorded and poorly known limno-terrestrial eutardigrade genus Apodibius have no claws on their strongly reduced legs, a unique character among all tardigrades. This absence of all claws makes the systematic position of Apodibius one of the most enigmatic among the whole class. Until now all known associates of the genus Apodibius have been located in the incertae sedis species group or, quite recently, included into the Necopinatidae family. In the present study, phylogenetic analyses of 18S and 28S rRNA sequence data from 31 tardigrade species representing four parachelan superfamilies (Isohypsibioidea, Hypsibioidea, Macrobiotoidea, Eohypsibioidea), the apochelan Milnesium tardigradum, and the type species of the genus Apodibius, A. confusus, indicated close relationship of the Apodibius with tardigrade species recently included in the superfamily Isohypsibioidea. This result was well-supported and consistent across all markers (separate 18S rRNA, 28S rRNA, and combined 18S rRNA+28S rRNA datasets) and methods (MP, ML) applied.

DNA barcodes reveal female dimorphism in syringophilid mites (Actinotrichida: Prostigmata: Cheyletoidea): Stibarokris phoeniconaias and Ciconichenophilus phoeniconaias are conspecific.

Here we present the first evidence of female dimorphism in ectoparasitic quill mites of the family Syringophilidae (Actinotrichida: Prostigmata: Cheyletoidea). Stibarokris phoeniconaias Skoracki et OConnor, 2010 and Ciconichenophilus phoeniconaias Skoracki et OConnor, 2010 so far have been treated as two distinct species cohabiting inside the quills of feathers of the lesser flamingo Phoeniconaias minor (Geoffroy Saint-Hilaire) and the American flamingo Phoenicopterus ruber Linnaeus. Although females of these species differ morphologically by the extent of body sclerotisation, presence/absence of lateral hypostomal teeth, and shape of dorsal setae, their important common features are the lack of leg setae vs II, and both stylophore and peritremes shape. Here, we apply the DNA barcode markers to test whether the differences between S. phoeniconaias and C. phoeniconaias have a genetic basis, indicating that they really are distinct taxa, or whether they just represent two morphs of a single species. All analysed sequences (616 bp for COI and 1159 bp for 28S rDNA) obtained for specimens representing females of both studied taxa as well as male, tritonymph, protonymph and larva of S. phoeniconaias were identical, which indicates that S. phoeniconaias and C. phoeniconaias are conspecific. The formal taxonomic consequence of our results is denial of the genus status of Ciconichenophilus Skoracki et OConnor, 2010 and species status of C. phoeniconaias, and recommendation that they should be treated as junior synonyms of Stibarokris Kethley, 1970 and S. phoeniconaias, respectively.

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.

Multidisciplinary analysis of Knemidocoptes jamaicensis parasitising the Common Chaffinch, Fringilla coelebs: proofs for a multispecies complex?

The number of studies discussing the pathology and host specificity in Knemidocoptinae is very limited. In Knemidocoptes jamaicensis, the host specificity seems to be very broad, and there is a clear morphological variability in individuals originating from various bird species; hence, serious doubts appear about the species status of this mite. We report a multidisciplinary approach to the taxonomy, morphology, ecology, and pathology of K. jamaicensis. The source of the mites in our study was a second year aged female of the Common Chaffinch, Fringilla coelebs, which accidentally died in the mist net during a field study in Dumbrava, Cluj County, Romania in March 2011. Comparisons of the biometrical data regarding the body dimensions, length of certain setae, and distances between bases of dorsal setae with other published data showed a great variability of certain measurements between populations infecting various hosts and localities and sometimes even within single populations. Gross and histologic lesions consisted in severe bilateral orthokeratotic hyperkeratosis and epidermal spongiosis. Lesions also involved the skin of the joints. Skin inflammation was absent, and no lesions were noticed in the metatarsus bone. Following molecular analysis, the 518-base-long sequence differed from the published 18S rDNA in nine positions. Additionally, our paper reports for the first time the DNA barcode sequences of K. jamaicensis and, together with the synoptic analysis of host spectrum, geographical distribution and morphological variability it brings important evidences to sustain the hypothesis of multispecies complex for K. jamaicensis.

Madeira's ptyctimous mites (Acari, Oribatida).

In the material recently collected in Madeira, 16 species of ptyctimous mites have been found. A new species of Austrophthiracarus rabacalensis Niedbala sp. nov. has been described. The presence of P. globosus and S. (R.) ortizi, reported earlier from Madeira has not been confirmed, but P. anonymus and P. montanus, so far not reported from this island, have been found. All 16 species identified in the material from Madeira studied occur in the Palaearctic Region; four of them are endemites, seven occur in western Palaearctic, four are panpalaearctic, while one is a semicosmopolitan. Morphological analysis has revealed a high similarity of two endemic species of Madeira with two European species: Steganacarus (Steganacarus) crassisetosus is similar to Steganacarus (Steganacarus) applicatus, while Steganacarus (Steganacarus) similis to Steganacarus (Steganacarus) spinosus. DNA-barcode analysis using COI and D2 28S rDNA sequences confirmed the species status of these four species. The phylogenetic analyses of COI amino acid data and D2 28S rDNA sequences suggest a closer relationship between S. (S.) crassisetosus and S. (S.) applicatus, pointing to a great genetic distance between S. (S.) spinosus and the other species of Steganacarus (Steganacarus).