Differential allergen expression in three Tyrophagus putrescentiae strains inhabited by distinct microbiome

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Populations of Stored Product Mite Tyrophagus putrescentiae Differ in Their Bacterial Communities

Citation: Erban, T., Klimov, P. B., Smrz, J., Phillips, T. W., Nesvorna, M., Kopecky, J., & Hubert, J. (2016). Populations of Stored Product Mite Tyrophagus putrescentiae Differ in Their Bacterial Communities. Frontiers in Microbiology, 7, 19. doi:10.3389/fmich.2015.01046Background: 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), Cardiniurn (five populations), Bartonella-like (five populations), Blattabacteriurn-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 Microbacteriurn. 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

Two Populations of Mites (Tyrophagus putrescentiae) Differ in Response to Feeding on Feces-Containing Diets

Background:Tyrophagus putrescentiae is a ubiquitous mite species in soil, stored products and house dust and infests food and causes allergies in people. T. putrescentiae populations harbor different bacterial communities, including intracellular symbionts and gut bacteria. The spread of microorganisms via the fecal pellets of T. putrescentiae is a possibility that has not been studied in detail but may be an important means by which gut bacteria colonize subsequent generations of mites. Feces in soil may be a vector for the spread of microorganisms.Methods: Extracts from used mite culture medium (i.e., residual food, mite feces, and dead mite bodies) were used as a source of feces-inhabiting microorganisms as food for the mites. Two T. putrescentiae populations (L and P) were used for experiments, and they hosted the intracellular bacteria Cardinium and Wolbachia, respectively. The effects of the fecal fraction on respiration in a mite microcosm, mite nutrient contents, population growth and microbiome composition were evaluated.Results: Feces from the P population comprised more than 90% Bartonella-like sequences. Feces from the L population feces hosted Staphylococcus, Virgibacillus, Brevibacterium, Enterobacteriaceae, and Bacillus. The mites from the P population, but not the L population, exhibited increased bacterial respiration in the microcosms in comparison to no-mite controls. Both L- and P-feces extracts had an inhibitory effect on the respiration of the microcosms, indicating antagonistic interactions within feces-associated bacteria. The mite microbiomes were resistant to the acquisition of new bacterial species from the feces, but their bacterial profiles were affected. Feeding of P mites on P-feces-enriched diets resulted in an increase in Bartonella abundance from 6 to 20% of the total bacterial sequences and a decrease in Bacillus abundance. The population growth was fivefold accelerated on P-feces extracts in comparison to the control.Conclusion: The mite microbiome, to a certain extent, resists the acquisition of new bacteria when mites are fed on feces of the same species. However, a Bartonella-like bacteria-feces-enriched diet seems to be beneficial for mite populations with symbiotic Bartonella-like bacteria. Coprophagy on the feces of its own population may be a mechanism of bacterial acquisition in T. putrescentiae

The effect of antibiotics on associated bacterial community of stored product mites.

Bacteria are associated with the gut, fat bodies and reproductive organs of stored product mites (Acari: Astigmata). The mites are pests due to the production of allergens. Addition of antibiotics to diets can help to characterize the association between mites and bacteria.Ampicillin, neomycin and streptomycin were added to the diets of mites and the effects on mite population growth (Acarus siro, Lepidoglyphus destructor and Tyrophagus putrescentiae) and associated bacterial community structure were assessed. Mites were treated by antibiotic supplementation (1 mg g(-1) of diet) for 21 days and numbers of mites and bacterial communities were analyzed and compared to the untreated control. Bacterial quantities, determined by real-time PCR, significantly decreased in antibiotic treated specimens from 5 to 30 times in A. siro and T. putrescentiae, while no decline was observed in L. destructor. Streptomycin treatment eliminated Bartonella-like bacteria in the both A. siro and T. putrescentiae and Cardinium in T. putrescentiae. Solitalea-like bacteria proportion increased in the communities of neomycin and streptomycin treated A. siro specimens. Kocuria proportion increased in the bacterial communities of ampicillin and streptomycin treated A. siro and neomycin and streptomycin treated L. destructor.The work demonstrated the changes of mite associated bacterial community under antibiotic pressure in pests of medical importance. Pre-treatment of mites by 1 mg g(-1) antibiotic diets improved mite fitness as indicated accelerated population growth of A. siro pretreated streptomycin and neomycin and L. destructor pretreated by neomycin. All tested antibiotics supplemented to diets caused the decrease of mite growth rate in comparison to the control diet

Detection and localization of Solitalea-like and Cardinium bacteria in three Acarus siro populations (Astigmata: Acaridae)

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. 1009 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 popu- lations 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

Cardinium endosymbionts are widespread in synanthropic mite species(Acari: Astigmata)

‘Candidatus Cardinium’ is an intracellular endosymbiont or parasite frequently occurring in invertebrates including mites and ticks. In this work we report Cardinium bacteria in Astigmata mites and explore their incidence in synanthropic species. Amplification of a 776 bp bacterial 16S rRNA gene fragment, using specific primers, enabled identification of closely related Cardinium sequences in 13 laboratory-reared populations of mites. In addition, Cardinium sequences were identified in three wild mite populations. Large scale screening of these populations showed 100% prevalence of Cardinium, representing the highest incidence compared to other major Chelicerate groups

The analyses of 16S rRNA libraries from the clones from stored product mites reared on control diets.

<p>The data from the three samples per treatment are presented altogether.</p><p>Legend: Accession number of match in GENBANK for the most similar sequences of identified bacteria I (%) describes the similarity; A– ampicillin treated diet, C – control diet, N – neomycin treated diet, S – streptomycin treated diet (all 1 mgg−1 of diet).</p><p>The analyses of 16S rRNA libraries from the clones from stored product mites reared on control diets.</p

Quantitative real-time PCR of total bacteria from the DNA extracted from stored product mites reared on control and antibiotic treated diets.

<p>The numbers of copies were recalculated per one specimen, averages with respective standard deviations. The Dunnet significant differences from the control are marked by asterisk; CON - untreated control, AMP –ampicillin, NEO – neomycin, STR – streptomycin.</p