Mites of the genus Tyrophagus (Acari: Acaridae) are among the most widespread and common mites, inhabiting diverse natural and anthropogenic habitats. Some species are pests of agricultural products and stored food and/or live in house dust, causing allergies to humans. We sequenced 1.2 kb of the mitochondrial COI gene for 38 individuals belonging to seven species of Tyrophagus, including T. curvipenis, T. putrescentiae, T. fanetzhangorum, T. longior, T. perniciosus, and T. cf. similis. Molecular phylogenetic analyses (1) recovered two major clades corresponding to the presence or absence of eyespots, and (2) separated all included morphological species. Tyrophagus curvipenis and T. putrescentiae had the lowest between-species genetic distances (range, mean ± SD): 14.20-16.30, 15.17 ± 0.40 (K2P). The highest within-species variation was found in T. putrescentiae 0.00-4.33, 1.78 ± 1.44 (K2P). In this species, we recovered two distinct groups; however, no geographical or ecological dissimilarities were observed between them. Based on our analyses, we document important morphological differences between T. curvipenis and T. putrescentiae. For the first time, we record the occurrence of T. curvipenis in the New World and suggest that it may be an emerging pest as it is currently spreading in agricultural produce.

• Klíčová slova
• COI, DNA barcode, Genetic distances, Tyrophagus curvipenis, Tyrophagus phylogeny,
• MeSH
• Acaridae anatomie a histologie   klasifikace   enzymologie   genetika   MeSH
• biologická evoluce * MeSH
• fylogeneze * MeSH
• mitochondriální proteiny genetika   MeSH
• proteiny členovců genetika   MeSH
• respirační komplex IV genetika   MeSH
• rozšíření zvířat MeSH
• sekvenční analýza DNA MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• mitochondriální proteiny MeSH
• proteiny členovců MeSH
• respirační komplex IV MeSH

Bacteria associated with mites influence their fitness, nutrition and reproduction. Previously, we found Solitalea-like (Sphingobacteriales) and Candidatus Cardinium (Cytophagales) bacteria in the stored product mite Acarus siro L. by cloning and using pyrosequencing. In this study, taxon-specific primers targeting 16S rRNA gene were used to detect and quantify the bacteria in mites and eggs of three A. siro populations. The specific probes for fluorescent in situ hybridization (FISH) were used to localize Solitalea-like and Cardinium bacteria in mite bodies. The population growth as an indirect estimator of fitness was used to describe the mite-bacteria interactions on (1) control diet; (2) rifampicin supplemented diet; (3) tetracycline supplemented diet; (4) rifampicin pretreated mites; (5) tetracycline pretreated mites. Solitalea-like 16S rRNA gene sequences from A. siro formed a separate cluster together with sequences from Tyrophagus putrescentiae. qPCR analysis indicated that number of Solitalea-like bacteria 16S rRNA gene copies was ca. 100× higher than that of Cardinium and the numbers differed between populations. FISH analysis localized Solitalea-like bacteria in the parenchymal tissues, mesodeum and food bolus of larvae, nymphs and adults. Solitalea-like, but not Cardinium bacteria were detected by taxon-specific primers in mites and eggs of all three investigated populations. None of the antibiotic treatments eliminated Solitalea-like bacteria in the A. siro populations tested. Rifampicin pretreatment significantly decreased the population growth. The numbers of Solitalea-like bacteria did not correlate with the population growth as a fitness indicator. This study demonstrated that A. siro can host Solitalea-like bacteria either alone or together with Cardinium. We suggest that Solitalea-like bacteria are shared by vertical transfer in A. siro populations.

• Klíčová slova
• Antibiotic treatment, Stored product mites, Symbiont, Transmission of bacteria,
• MeSH
• Acaridae růst a vývoj   mikrobiologie   MeSH
• antibakteriální látky farmakologie   MeSH
• Bacteroidetes klasifikace   genetika   fyziologie   MeSH
• DNA bakterií genetika   MeSH
• genetická zdatnost * MeSH
• hybridizace in situ fluorescenční MeSH
• krmivo pro zvířata analýza   MeSH
• larva růst a vývoj   mikrobiologie   MeSH
• nymfa růst a vývoj   mikrobiologie   MeSH
• RNA ribozomální 16S genetika   MeSH
• symbióza MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antibakteriální látky MeSH
• DNA bakterií MeSH
• RNA ribozomální 16S MeSH

BACKGROUND: Tyrophagus putrescentiae colonizes different human-related habitats and feeds on various post-harvest foods. The microbiota acquired by these mites can influence the nutritional plasticity in different populations. We compared the bacterial communities of five populations of T. putrescentiae and one mixed population of T. putrescentiae and T. fanetzhangorum collected from different habitats. MATERIAL: The bacterial communities of the six mite populations from different habitats and diets were compared by Sanger sequencing of cloned 16S rRNA obtained from amplification with universal eubacterial primers and using bacterial taxon-specific primers on the samples of adults/juveniles or eggs. Microscopic techniques were used to localize bacteria in food boli and mite bodies. The morphological determination of the mite populations was confirmed by analyses of CO1 and ITS fragment genes. RESULTS: The following symbiotic bacteria were found in compared mite populations: Wolbachia (two populations), Cardinium (five populations), Bartonella-like (five populations), Blattabacterium-like symbiont (three populations), and Solitalea-like (six populations). From 35 identified OTUs97, only Solitalea was identified in all populations. The next most frequent and abundant sequences were Bacillus, Moraxella, Staphylococcus, Kocuria, and Microbacterium. We suggest that some bacterial species may occasionally be ingested with food. The bacteriocytes were observed in some individuals in all mite populations. Bacteria were not visualized in food boli by staining, but bacteria were found by histological means in ovaria of Wolbachia-infested populations. CONCLUSION: The presence of Blattabacterium-like, Cardinium, Wolbachia, and Solitalea-like in the eggs of T. putrescentiae indicates mother to offspring (vertical) transmission. RESULTS of this study indicate that diet and habitats influence not only the ingested bacteria but also the symbiotic bacteria of T. putrescentiae.

• Klíčová slova
• 16S rRNA, Blattabacterium, Tyrophagus putrescentiae, Wolbachia, bacteria, feeding, symbiont,
• Publikační typ
• časopisecké články MeSH

Tyrophagus putrescentiae (Schrank, 1781) is an emerging source of allergens in stored products and homes. Feces proteases are the major allergens of astigmatid mites (Acari: Acaridida). In addition, the mites are carriers of microorganisms and microbial adjuvant compounds that stimulate innate signaling pathways. We sought to analyze the mite feces proteome, proteolytic activities, and mite-bacterial interaction in dry dog food (DDF). Proteomic methods comprising enzymatic and zymographic analysis of proteases and 2D-E-MS/MS were performed. The highest protease activity was assigned to trypsin-like proteases; lower activity was assigned to chymotrypsin-like proteases, and the cysteine protease cathepsin B-like had very low activity. The 2D-E-MS/MS proteomic analysis identified mite trypsin allergen Tyr p3, fatty acid-binding protein Tyr p13 and putative mite allergens ferritin (Grp 30) and (poly)ubiquitins. Tyr p3 was detected at different positions of the 2D-E. It indicates presence of zymogen at basic pI, and mature-enzyme form and enzyme fragment at acidic pI. Bacillolysins (neutral and alkaline proteases) of Bacillus cereus symbiont can contribute to the protease activity of the mite extract. The bacterial exo-chitinases likely contribute to degradation of mite exuviae, mite bodies or food boluses consisting of chitin, including the peritrophic membrane. Thus, the chitinases disrupt the feces and facilitate release of the allergens. B. cereus was isolated and identified based on amplification and sequencing of 16S rRNA and motB genes. B. cereus was added into high-fat, high-protein (DDF) and low-fat, low-protein (flour) diets to 1 and 5% (w/w), and the diets palatability was evaluated in 21-day population growth test. The supplementation of diet with B. cereus significantly suppressed population growth and the suppressive effect was higher in the high-fat, high-protein diet than in the low-fat, low-protein food. Thus, B. cereus has to coexist with the mite in balance to be beneficial for the mite. The mite-B. cereus symbiosis can be beneficial-suppressive at some level. The results increase the veterinary and medical importance of the allergens detected in feces. The B. cereus enzymes/toxins are important components of mite allergens. The strong symbiotic association of T. putrescentiae with B. cereus in DDF was indicated.

• Klíčová slova
• Bacillus cereus, Tyrophagus putrescentiae, allergen, bacillolysin, exochitinase, nutrition, protease, symbiosis,
• Publikační typ
• časopisecké články MeSH

The rate of population increase of three mite species, Acarus siro (L.), Lepidoglyphus destructor (Schrank) and Tyrophagus putrescentiae (Schrank), was studied on various types of barley and at various combinations of temperature and humidity. The mites were added into the chambers and incubated for 21 days on seven different kinds of barley coming from four sites, including six cultivars and a mixture. The population increase of all species was higher on the mixture than on any other cultivar, except for Sebastian and Calgary. The increase of mites was studied at constant temperatures ranging from 5 to 35 °C and relative humidity (RH) ranging from 50 to 90 %. Positive rate of increase was found above 70 % RH for all species. The optimal humidity was at 85 % RH for A. siro and L. destructor and at 90 % RH for T. putrescentiae. As concerns the temperature, positive rate of increase was found at temperatures higher than 10, 15 and 20 °C for A. siro, L. destructor and T. putrescentiae, respectively. The temperature optima were at 23, 25, and 30 °C for A. siro, L. destructor and T. putrescentiae, respectively. Model estimated on laboratory data was then fitted to temperature and humidity records from August to November in the Czech grain store. Estimated population rate of increase was rarely positive: for A. siro it was for 24 %, for L. destructor for only 1 % and for T. putrescentiae for only 7 % days of the study period. It is concluded that in the climatic conditions of the Czech Republic the population increase of three mite pests is negligible during autumn and winter.

• MeSH
• Acari fyziologie   MeSH
• ječmen (rod) klasifikace   MeSH
• populační dynamika MeSH
• teplota * MeSH
• vlhkost * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Tyrophagus putrescentiae (Acari: Astigmata) and Fusarium sp. co-occur in poorly managed grain. In a laboratory experiment, mite grazing resulted in significant reduction of fungal mycelium on cultivation plates. The destruction of mycelium appeared to be a result of an interaction between the mites, fungi and associated bacteria. METHODOLOGY AND PRINCIPAL FINDINGS: A laboratory experiment was performed to simulate a situation of grain multiinfested by mites and Fusarium fungi. Changes of mite-associated bacterial community in T. putrescentiae were described in 3 habitats: (i) T. putrescentiae mites from a rearing diet prior to their transfer to fungal diet; (ii) fungal mycelium before mite introduction; (iii) mites after 7 day diet of each Fusarium avenaceum, F. culmorum, F. poae and F. verticillioides. Bacterial communities were characterized by 16 S rRNA gene sequencing. In total, 157 nearly full-length 16 S rRNA gene sequences from 9 samples representing selected habitats were analyzed. In the mites, the shift from rearing to fungal diet caused changes in mite associated bacterial community. A diverse bacterial community was associated with mites feeding on F. avenaceum, while feeding on the other three Fusarium spp. led to selection of a community dominated by Bacillaceae. CONCLUSIONS/SIGNIFICANCE: The work demonstrated changes of bacterial community associated with T. putrescentiae after shift to fungal diets suggesting selection for Bacillaceae species known as chitinase producers, which might participate in the fungal mycelium hydrolysis.

• MeSH
• Acaridae mikrobiologie   MeSH
• Bacteria klasifikace   MeSH
• dieta * MeSH
• druhová specificita MeSH
• Fusarium růst a vývoj   MeSH
• jedlá semena mikrobiologie   MeSH
• kultivační techniky MeSH
• mycelium růst a vývoj   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Internal bacterial communities of synanthropic mites Acarus siro, Dermatophagoides farinae, Lepidoglyphus destructor, and Tyrophagus putrescentiae (Acari: Astigmata) were analyzed by culturing and culture-independent approaches from specimens obtained from laboratory colonies. Homogenates of surface-sterilized mites were used for cultivation on non-selective agar and DNA extraction. Isolated bacteria were identified by sequencing of the 16S rRNA gene. PCR amplified 16S rRNA genes were analyzed by terminal restriction fragment length polymorphism analysis (T-RFLP) and cloning sequencing. Fluorescence in situ hybridization using universal bacterial probes was used for direct bacterial localization. T-RFLP analysis of 16S rRNA gene revealed distinct species-specific bacterial communities. The results were further confirmed by cloning and sequencing (284 clones). L. destructor and D. farinae showed more diverse communities then A. siro and T. putrescentiae. In the cultivated part of the community, the mean CFUs from four mite species ranged from 5.2 × 10(2) to 1.4 × 10(3) per mite. D. farinae had significantly higher CFUs than the other species. Bacteria were located in the digestive and reproductive tract, parenchymatical tissue, and in bacteriocytes. Among the clones, Bartonella-like bacteria occurring in A. siro and T. putresecentiae represented a distinct group related to Bartonellaceae and to Bartonella-like symbionts of ants. The clones of high similarity to Xenorhabdus cabanillasii were found in L. destructor and D. farinae, and one clone related to Photorhabdus temperata in A. siro. Members of Sphingobacteriales cloned from D. farinae and A. siro clustered with the sequences of "Candidatus Cardinium hertigii" and as a separate novel cluster.

• MeSH
• Acari klasifikace   genetika   mikrobiologie   MeSH
• Bacteria klasifikace   genetika   izolace a purifikace   MeSH
• Dermatophagoides farinae mikrobiologie   MeSH
• druhová specificita MeSH
• hybridizace in situ fluorescenční MeSH
• klonování DNA MeSH
• molekulární sekvence - údaje MeSH
• polymerázová řetězová reakce MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• RNA ribozomální 16S genetika   MeSH
• roztoči mikrobiologie   MeSH
• sekvenční analýza DNA MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• RNA ribozomální 16S MeSH

Bacillus thuringiensis (Bt) toxins present a potential for control of pest mites. Information concerning the effect of Bt and its possible application to the biocontrol of synathropic mites is rare. The toxic effect of Bacillus thuringiensis var. tenebrionis producing Cry3A toxin was tested on the mites Acarus siro L., Tyrophagus putrescentiae (Schrank), Dermatophagoides farinae Hughes, and Lepidoglyphus destructor (Schrank) via feeding tests. Fifty mites were reared on Bt additive diets in concentrations that ranged from 0 to 100 mg g(-1) under optimal conditions for their development. After 21 days, the mites were counted and the final populations were analyzed using a polynomial regression model. The Bt diet suppressed population growth of the four mite species. The fitted doses of Bt for 50% suppression of population growth were diets ranging from 25 to 38 mg g(-1). There were no remarkable differences among species. Possible applications of Bt for the control of synanthropic mites are discussed.

• MeSH
• akaricidy * MeSH
• bakteriální proteiny * MeSH
• biologická kontrola škůdců * MeSH
• druhová specificita MeSH
• endotoxiny * MeSH
• hemolyziny * MeSH
• roztoči * MeSH
• toxiny Bacillus thuringensis MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• akaricidy * MeSH
• bakteriální proteiny * MeSH
• endotoxiny * MeSH
• hemolyziny * MeSH
• insecticidal crystal protein, Bacillus Thuringiensis MeSH Prohlížeč
• toxiny Bacillus thuringensis MeSH

Eggs of 13 species of common, economically important stored-product pest mites (Acarus siro, A. gracilis, Tyrophagus putrescentiae, T. brevicrinatus, Tyroborus lini, Aleuroglyphus ovatus, Caloglyphus redikorzevi, C. oudemansi, Lepidoglyphus destructor, Glycyphagus domesticus, Aeroglyphus robustus, Chortoglyphus arcuatus and Carpoglyphus lactis) from four families (Acaridae, Glycyphagidae, Chortoglyphidae, Carpoglyphidae) were studied in order to build a diagnostic key. The morphological study dealt with shape, size and surface sculpturing of eggs. Morphological details were visualised using scanning electron microscopy. A key for distinguishing eggs at genus and species level was developed for the major stored-product mites.

• MeSH
• Acaridae klasifikace   cytologie   ultrastruktura   MeSH
• kontrola škůdců MeSH
• mikroskopie elektronová rastrovací MeSH
• ovum klasifikace   cytologie   ultrastruktura   MeSH
• povrchové vlastnosti MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The toxicities of three plant volatiles, (2E)-hexenal, (2E, 6Z)-nonadienal and (2E)-nonenal, intermediate products of the oxylipin biosynthesis pathway, were tested on three mites of importance for medical purposes and as pests. The aldehydes were diluted in hexane separately and incorporated into diets in ranges of 4-143 mg g(-1). The final density of mites in control and aldehyde-enriched diets was compared after 21 days. The aldehydes were toxic to the mites, whose final density showed an inverse correlation with aldehyde concentration. In addition to the effects of aldehyde concentration, the final density of mites was also influenced by the different aldehydes tested and the interaction among aldehyde concentration and chemical structure. In a functional combination of aldehydes and species, the doses calculated for growth inhibition and eradication of mites ranged from 4 to 35 mg g(-1) and from 36 to 314 mg g(-1), respectively. Due to the protective role displayed by natural six-carbon and nine-carbon aldehydes, these compounds are potential candidates for controlling stored-product mites in stored food and feed products.

• MeSH
• Acaridae účinky léků   MeSH
• aldehydy farmakologie   MeSH
• druhová specificita MeSH
• insekticidy farmakologie   MeSH
• roztoči účinky léků   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aldehydy MeSH
• insekticidy MeSH