The Effect of Residual Pesticide Application on Microbiomes of the Storage Mite Tyrophagus putrescentiae.

Arthropods can host well-developed microbial communities, and such microbes can degrade pesticides and confer tolerance to most types of pests. Two cultures of the stored-product mite Tyrophagus putrescentiae, one with a symbiotic microbiome containing Wolbachia and the other without Wolbachia, were compared on pesticide residue (organophosphate: pirimiphos-methyl and pyrethroid: deltamethrin, deltamethrin + piperonyl butoxide)-containing diets. The microbiomes from mite bodies, mite feces and debris from the spent mite diet were analyzed using barcode sequencing. Pesticide tolerance was different among mite cultures and organophosphate and pyrethroid pesticides. The pesticide residues influenced the microbiome composition in both cultures but without any remarkable trend for mite cultures with and without Wolbachia. The most influenced bacterial taxa were Bartonella-like and Bacillus for both cultures and Wolbachia for the culture containing this symbiont. However, there was no direct evidence of any effect of Wolbachia on pesticide tolerance. The high pesticide concentration residues in diets reduced Wolbachia, Bartonella-like and Bacillus in mites of the symbiotic culture. This effect was low for Bartonella-like and Bacillus in the asymbiotic microbiome culture. The results showed that the microbiomes of mites are affected by pesticide residues in the diets, but the effect is not systemic. No actual detoxification effect by the microbiome was observed for the tested pesticides.

Pesticide residue exposure provides different responses of the microbiomes of distinct cultures of the stored product pest mite Acarus siro.

BACKGROUND: The contribution of the microbiome to pesticide breakdown in agricultural pests remains unclear. We analyzed the effect of pirimiphos-methyl (PM) on four geographically different cultures of the stored product pest mite Acarus siro (6 L, 6Tu, 6Tk and 6Z) under laboratory experiments. The effect of PM on mite mortality in the impregnated filter paper test was compared. RESULTS: The mite sensitivity to PM decreased in the order of 6 L, 6Tu, 6Tk, and 6Z. Then, the mites were cultured on PM residues (0.0125 and 1.25 µg·g-1), and population growth was compared to the control after 21 days of exposure. The comparison showed two situations: (i) increasing population growth for the most sensitive cultures (6 L and 6Tu), and (ii) no effect on mite population growth for tolerant cultures (6Z and 6Tk). The microbiome of mites was analyzed by quantification of 16S DNA copies based on quantitative polymerase chain reaction (qPCR) and by barcode sequencing of the V4 fragment of 16S DNA on samples of 30 individuals from the control and PM residues. The microbiome comprised primarily Solitalea-like organisms in all cultures, except for 6Z, followed by Bacillus, Staphylococcus, and Lactobacillus. The microbiomes of mite cultures did not change with increasing population density. The microbiome of cultures without any differences in population density showed differences in the microbiome composition. A Sodalis-like symbiont replaced Solitalea in the 1.25 µg·g-1 PM in the 6Tk culture. Sodalis and Bacillus prevailed in the microbiomes of PM-treated mites of 6Z culture, while Solitalea was almost absent. CONCLUSION: The results showed that the microbiome of A. siro differs in composition and in response to PM residues in the diet. The results indicate that Sodalis-like symbionts can help recover mites from pesticide-induced stress.

Microbial Communities of Stored Product Mites: Variation by Species and Population.

Arthropod-associated microorganisms are important because they affect host fitness, protect hosts from pathogens, and influence the host's ability to vector pathogens. Stored product mites (Astigmata) often establish large populations in various types of food items, damaging the food by direct feeding and introducing contaminants, including their own bodies, allergen-containing feces, and associated microorganisms. Here we access the microbial structure and abundance in rearing diets, eggs, feces fraction, and mite bodies of 16 mite populations belonging to three species (Carpoglyphus lactis, Acarus siro, and Tyrophagus putrescentiae) using quantitative PCR and 16S ribosomal RNA (rRNA) gene amplicon sequencing. The mite microbiomes had a complex structure dominated by the following bacterial taxa (OTUs): (a) intracellular symbionts of the genera Cardinium and Wolbachia in the mite bodies and eggs; (b) putative gut symbionts of the genera Solitalea, Bartonella, and Sodalis abundant in mite bodies and also present in mite feces; (c) feces-associated or environmental bacteria of the genera Bacillus, Staphylococcus, and Kocuria in the diet, mite bodies, and feces. Interestingly and counterintuitively, the differences between microbial communities in various conspecific mite populations were higher than those between different mite species. To explain some of these differences, we hypothesize that the intracellular bacterial symbionts can affect microbiome composition in mite bodies, causing differences between microbial profiles. Microbial profiles differed between various sample types, such as mite eggs, bodies, and the environment (spent growth medium-SPGM). Low bacterial abundances in eggs may result in stochastic effects in parent-offspring microbial transmission, except for the intracellular symbionts. Bacteria in the rearing diet had little effect on the microbial community structure in SPGM and mite bodies. Mite fitness was positively correlated with bacterial abundance in SPGM and negatively correlated with bacterial abundances in mite bodies. Our study demonstrates critical host-microbe interactions, affecting all stages of mite growth and leading to alteration of the environmental microbiome. Correlational evidence based on absolute quantitation of bacterial 16S rRNA gene copies suggests that mite-associated microorganisms are critical for modulating important pest properties of mites by altering population growth.

The Mite Tyrophagus putrescentiae Hosts Population-Specific Microbiomes That Respond Weakly to Starvation.

The effect of short-term nutrient deprivation was studied in five populations of the mite Tyrophagus putrescentiae with different microbiomes. The fresh weight, nutrient status, respiration, and population growth of the mites were observed for the five mite population-scale samples. The starvation caused the larvae and nymphs to be eliminated, resulting in a significant increase in the fresh weight of starved adult specimens. Three populations were negatively influenced by starvation, and the starved specimens were characterized by a decrease in nutrient status, respiration, and population growth. One population was not influenced or was slightly influenced by starvation, which had no effect on population growth or nutrient contents but caused a significant decrease in respiration. One population was positively influenced by starvation; the population growth increased in starved specimens, and starvation had no effect on respiration. Although starvation altered the bacterial profiles of the microbiomes, these differences were much smaller than those between the populations. The bacterial profiles of Staphylococcus, Bacillus, Kocuria, Brevibacterium, and unidentified Micrococcaceae and Enterobacteriaceae increased in starved specimens, whereas those of Bartonella and Solitalea-like genera were reduced in the starved mite populations. The profiles of the intracellular symbiont Cardinium decreased in the starved specimens, and the Wolbachia profile changes were dependent on the mite population. In mite populations, when the symbionts were rare, their profiles varied stochastically. Correlations between changes in the profiles of the bacterial taxa and mite fitness parameters, including nutrient status (lipids, proteins, saccharides, and glycogen contents), mite population growth, and respiration, were observed. Although the microbiomes were resistant to the perturbations caused by nutrition deficiency, the responses of the mites differed in terms of their population growth, respiration, and nutrient status.

Population and Culture Age Influence the Microbiome Profiles of House Dust Mites.

Interactions with microorganisms might enable house dust mites (HDMs) to derive nutrients from difficult-to-digest structural proteins and to flourish in human houses. We tested this hypothesis by investigating the effects of changes in the mite culture growth and population of two HDM species on HDM microbiome composition and fitness. Growing cultures of laboratory and industrial allergen-producing populations of Dermatophagoides farinae (DFL and DFT, respectively) and Dermatophagoides pteronyssinus (DPL and DPT, respectively) were sampled at four time points. The symbiotic microorganisms of the mites were characterized by DNA barcode sequencing and quantified by qPCR using universal/specific primers. The population growth of mites and nutrient contents of mite bodies were measured and correlated with the changes in bacteria in the HDM microbiome. The results showed that both the population and culture age significantly influenced the microbiome profiles. Cardinium formed 93% and 32% of the total sequences of the DFL and DFT bacterial microbiomes, respectively, but this bacterial species was less abundant in the DPL and DPT microbiomes. Staphylococcus abundance was positively correlated with increased glycogen contents in the bodies of mites, and increased abundances of Aspergillus, Candida, and Kocuria were correlated with increased lipid contents in the bodies of mites. The xerophilic fungus Wallemia accounted for 39% of the fungal sequences in the DPL microbiome, but its abundance was low in the DPT, DFL, and DFT microbiomes. With respect to the mite culture age, we made three important observations: the mite population growth from young cultures was 5-8-fold higher than that from old cultures; specimens from old cultures had greater abundances of fungi and bacteria in their bodies; and yeasts predominated in the gut contents of specimens from young cultures, whereas filamentous mycelium prevailed in specimens from old cultures. Our results are consistent with the hypothesis that mites derive nutrients through associations with microorganisms.

Detection of tau-fluvalinate resistance in the mite Varroa destructor based on the comparison of vial test and PCR-RFLP of kdr mutation in sodium channel gene.

Varroa destructor is the major cause of honey bee (Apis mellifera) colony losses. Mite control is limited to several miticides. The overuse of tau-fluvalinate has resulted in resistance via a knockdown resistance (kdr) mutation in the sodium channel gene NaVChs (L925V/I/M). In this study, we used the discriminating concentration of tau-fluvalinate (0.25 µg/mL) to detect the resistance of mites in a bioassay. Further, we verified the presence of the kdr mutation in mites from the bioassay via PCR amplification of a fragment of the voltage-gated sodium channel gene (NaVCh), restriction fragment length polymorphisms (RFLPs), and densitometry analyses in pools of surviving or dead mites. Resistance values corresponding to the densitometry of the resistant allele were related to mite survival. In the vial test, the survival of the control group was significantly higher (70.4%) than that of the tau-fluvalinate-treated group (34.3%). Mite survival in the vial test was significantly correlated with the mean proportion of resistance values. Individuals that died after tau-fluvalinate application exhibited an average resistance value of 0.0783, whereas individuals that survived exhibited an average resistance of 0.400. The concentration of tau-fluvalinate in the vials was checked using high performance liquid chromatography under different temperatures and exposure times, and indicates that the stability of tau-fluvalinate stored in the refrigerator (4 ± 1 °C) is at least 14 days. PCR-RFLP of the NaVCh gene fragment verified that the vial test is a suitable, rapid, and cost-effective method for the identification of tau-fluvalinate resistance based on kdr mutation in V. destructor in apiaries.

Experimental Manipulation Shows a Greater Influence of Population than Dietary Perturbation on the Microbiome of Tyrophagus putrescentiae.

Tyrophagus putrescentiae is inhabited by bacteria that differ among mite populations (strains) and diets. Here, we investigated how the microbiome and fitness of Tputrescentiae are altered by dietary perturbations and mite populations. Four T. putrescentiae populations, referred to as dog, Koppert, laboratory, and Phillips, underwent a perturbation, i.e., a dietary switch from a rearing diet to two experimental diets. The microbiome was investigated by sequencing the V1-V3 portion of the 16S rRNA gene, and selected bacterial taxa were quantified by quantitative PCR (qPCR) using group/taxon-specific primers. The parameters observed were the changes in mite population growth and nutritional status, i.e., the total glycogen, lipid, saccharide, and protein contents in mites. The effect of diet perturbation on the variability of the microbiome composition and population growth was lower than the effect induced by mite population. In contrast, the diet perturbation showed a greater effect on nutritional status of mites than the mite population. The endosymbionts exhibited high variations among T. putrescentiae populations, including Cardinium in the laboratory population, Blattabacterium-like bacteria in the dog population, and Wolbachia in the dog and Phillips populations. Solitalea-like and Bartonella-like bacteria were present in the dog, Koppert, and Phillips populations in different proportions. The T. putrescentiae microbiome is dynamic and varies based on both the mite population and perturbation; however, the mites remain characterized by robust bacterial communities. Bacterial endosymbionts were found in all populations but represented a dominant portion of the microbiome in only some populations.IMPORTANCE We addressed the question of whether population origin or perturbation exerts a more significant influence on the bacterial community of the stored product mite Tyrophagus putrescentiae The microbiomes of four populations of T. putrescentiae insects subjected to diet perturbation were compared. Based on our results, the bacterial community was more affected by the mite population than by diet perturbation. This result can be interpreted as indicating high stability of the putative intracellular symbionts in response to dietary perturbation. The changes in the absolute and relative numbers of Wolbachia, Blattabacterium-like, Solitalea-like, and Cardinium bacteria in the T. putrescentiae populations can also be caused by neutral processes other than perturbation. When nutritional status is considered, the effect of population appeared less important than the perturbation. We hypothesize that differences in the proportions of the endosymbiotic bacteria result in changes in mite population growth.

Changes in the Bacteriome of Honey Bees Associated with the Parasite Varroa destructor, and Pathogens Nosema and Lotmaria passim.

The honey bee, Apis mellifera, is a globally important species that suffers from a variety of pathogens and parasites. These parasites and pathogens may have sublethal effects on their bee hosts via an array of mechanisms, including through a change in symbiotic bacterial taxa. Our aim was to assess the influence of four globally widespread parasites and pathogens on the honey bee bacteriome. We examined the effects of the ectoparasitic mite Varroa destructor, the fungal pathogens Nosema apis and Nosema ceranae, and the trypanosome Lotmaria passim. Varroa was detected by acaricidal treatment, Nosema and L. passim by PCR, and the bacteriome using MiSeq 16S rRNA gene sequencing. Overall, the 1,858,850 obtained sequences formed 86 operational taxonomic units (OTUs) at 3 % dissimilarity. Location, time of year, and degree of infestation by Varroa had significant effects on the composition of the bacteriome of honey bee workers. Based on statistical correlations, we found varroosis more important factor than N. ceranae, N. apis, and L. passim infestation influencing the honey bee bacteriome and contributing to the changes in the composition of the bacterial community in adult bees. At the population level, Varroa appeared to modify 20 OTUs. In the colonies with high Varroa infestation levels (varroosis), the relative abundance of the bacteria Bartonella apis and Lactobacillus apis decreased. In contrast, an increase in relative abundance was observed for several taxa including Lactobacillus helsingborgensis, Lactobacillus mellis, Commensalibacter intestini, and Snodgrassella alvi. The results showed that the "normal" bacterial community is altered by eukaryotic parasites as well as displaying temporal changes and changes associated with the geographical origin of the beehive.

Comparison of Microbiomes between Red Poultry Mite Populations (Dermanyssus gallinae): Predominance of Bartonella-like Bacteria.

Blood feeding red poultry mites (RPM) serve as vectors of pathogenic bacteria and viruses among vertebrate hosts including wild birds, poultry hens, mammals, and humans. The microbiome of RPM has not yet been studied by high-throughput sequencing. RPM eggs, larvae, and engorged adult/nymph samples obtained in four poultry houses in Czechia were used for microbiome analyses by Illumina amplicon sequencing of the 16S ribosomal RNA (rRNA) gene V4 region. A laboratory RPM population was used as positive control for transcriptome analysis by pyrosequencing with identification of sequences originating from bacteria. The samples of engorged adult/nymph stages had 100-fold more copies of 16S rRNA gene copies than the samples of eggs and larvae. The microbiome composition showed differences among the four poultry houses and among observed developmental stadia. In the adults' microbiome 10 OTUs comprised 90 to 99% of all sequences. Bartonella-like bacteria covered between 30 and 70% of sequences in RPM microbiome and 25% bacterial sequences in transcriptome. The phylogenetic analyses of 16S rRNA gene sequences revealed two distinct groups of Bartonella-like bacteria forming sister groups: (i) symbionts of ants; (ii) Bartonella genus. Cardinium, Wolbachia, and Rickettsiella sp. were found in the microbiomes of all tested stadia, while Spiroplasma eriocheiris and Wolbachia were identified in the laboratory RPM transcriptome. The microbiomes from eggs, larvae, and engorged adults/nymphs differed. Bartonella-like symbionts were found in all stadia and sampling sites. Bartonella-like bacteria was the most diversified group within the RPM microbiome. The presence of identified putative pathogenic bacteria is relevant with respect to human and animal health issues while the identification of symbiontic bacteria can lead to new control methods targeting them to destabilize the arthropod host.

Comparison of Varroa destructor and Worker Honeybee Microbiota Within Hives Indicates Shared Bacteria.

The ectoparasitic mite Varroa destructor is a major pest of the honeybee Apis mellifera. In a previous study, bacteria were found in the guts of mites collected from winter beehive debris and were identified using Sanger sequencing of their 16S rRNA genes. In this study, community comparison and diversity analyses were performed to examine the microbiota of honeybees and mites at the population level. The microbiota of the mites and honeybees in 26 colonies in seven apiaries in Czechia was studied. Between 10 and 50 Varroa females were collected from the bottom board, and 10 worker bees were removed from the peripheral comb of the same beehive. Both bees and mites were surface sterilized. Analysis of the 16S rRNA gene libraries revealed significant differences in the Varroa and honeybee microbiota. The Varroa microbiota was less diverse than was the honeybee microbiota, and the relative abundances of bacterial taxa in the mite and bee microbiota differed. The Varroa mites, but not the honeybees, were found to be inhabited by Diplorickettsia. The relative abundance of Arsenophonus, Morganella, Spiroplasma, Enterococcus, and Pseudomonas was higher in Varroa than in honeybees, and the Diplorickettsia symbiont detected in this study is specific to Varroa mites. The results demonstrated that there are shared bacteria between Varroa and honeybee populations but that these bacteria occur in different relative proportions in the honeybee and mite bacteriomes. These results support the suggestion of bacterial transfer via mites, although only some of the transferred bacteria may be harmful.

Comparison of tau-fluvalinate, acrinathrin, and amitraz effects on susceptible and resistant populations of Varroa destructor in a vial test.

The parasitic mite Varroa destructor is a major pest of the western honeybee, Apis mellifera. The development of acaricide resistance in Varroa populations is a global issue. Discriminating concentrations of acaricides are widely used to detect pest resistance. Two methods, using either glass vials or paraffin capsules, are used to screen for Varroa resistance to various acaricides. We found the glass vial method to be useless for testing Varroa resistance to acaridices, so we developed a polypropylene vial bioassay. This method was tested on tau-fluvalinate-, acrinathrin-, and amitraz-resistant mite populations from three apiaries in Czechia. Acetone was used as a control and technical grade acaricide compounds diluted in acetone were applied to the polypropylene vials. The solutions were spread on the vial surface by rolling the vial, and were then evaporated. Freshly collected Varroa females were placed in the vials and the mortality of the exposed mites was measured after 24 h. The Varroa populations differed in mortality between the apiaries and the tested compounds. Mites from the Kyvalka site were resistant to acrinathrin, tau-fluvalinate, and amitraz, while mites from the Postrizin site were susceptible to all three acaricides. In Prelovice apiary, the mites were susceptible to acrinathrin and amitraz, but not to tau-fluvalinate. The calculated discriminating concentrations for tau-fluvalinate, acrinathrin, and amitraz were 0.66, 0.26 and 0.19 µg/mL, respectively. These results indicate that polyproplyne vial tests can be used to determine discriminating concentrations for the early detection of acaricide resistant Varroa. Finally, multiple-resistance in Kyvalka may indicate metabolic resistance.

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. 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.

Prevalence of pathogenic bacteria in Ixodes ricinus ticks in Central Bohemia.

Bacteria associated with the tick Ixodes ricinus were assessed in specimens unattached or attached to the skin of cats, dogs and humans, collected in the Czech Republic. The bacteria were detected by PCR in 97 of 142 pooled samples including 204 ticks, i.e. 1-7 ticks per sample, collected at the same time from one host. A fragment of the bacterial 16S rRNA gene was amplified, cloned and sequenced from 32 randomly selected samples. The most frequent sequences were those related to Candidatus Midichloria midichlori (71% of cloned sequences), followed by Diplorickettsia (13%), Spiroplasma (3%), Rickettsia (3%), Pasteurella (3%), Morganella (3%), Pseudomonas (2%), Bacillus (1%), Methylobacterium (1%) and Phyllobacterium (1%). The phylogenetic analysis of Spiroplasma 16S rRNA gene sequences showed two groups related to Spiroplasma eriocheiris and Spiroplasma melliferum, respectively. Using group-specific primers, the following potentially pathogenic bacteria were detected: Borellia (in 20% of the 142 samples), Rickettsia (12%), Spiroplasma (5%), Diplorickettsia (5%) and Anaplasma (2%). In total, 68% of I. ricinus samples (97/142) contained detectable bacteria and 13% contained two or more putative pathogenic groups. The prevalence of tick-borne bacteria was similar to the observations in other European countries.

Stored product mites (Acari: Astigmata) infesting food in various types of packaging.

From 2008 to 2014, stored product mites have been reported from prepackaged dried food on the market in the Czech Republic. The infestation was by Carpoglyphus lactis (L.) in dried fruits and Tyrophagus putrescentiae (Schrank) in dog feed. The infestation is presumably caused by poor protection of the packages. We compared various packaging methods for their resistance to mites using dried apricots and dog feed in laboratory experiments. The trial packages included nine different plastic films, monofilm, duplex and triplex, and one type of plastic cup (ten replicates per packaging type). All packaging materials are available on the Czech market for dried food products. The samples of dried food were professionally packed in a factory and packaged dried apricots were exposed to C. lactis and dog food to T. putrescentiae. After 3 months of exposure, the infestation and mite density of the prepackaged food was assessed. Mites were found to infest six types of packages. Of the packaging types with mites, 1-5 samples were infested and the maximum abundance was 1,900 mites g(-1) of dried food. Mites entered the prepackaged food by faulty sealing. Inadequate sealing is suggested to be the major cause of the emerged infestation of dried food.

Growth-suppressive effect of the α-amylase inhibitor of Triticum aestivum on stored-product mites varies by the species and type of diet.

A naturally occurring α-amylase inhibitor (α-AI) of Triticum aestivum protects wheat grain from gramnivorous arthropod pests. The α-AI (Type-I) was incorporated into carbohydrate and protein diets to test its inhibitory activity on the stored-product mites Acarus siro, Lepidoglyphus destructor and Tyrophagus putrescentiae (Acari: Astigmata). Growth tests of mites fed the various diets were used to compare the suppressive effects. The final population size of mites attained from an initial population of 50 specimens maintained under controlled conditions (85 % relative humidity and 25 °C) was compared after 21 days of cultivation. The results showed that α-AI in the concentration in the range of 0.01-1 mg g(-1) did not suppress the growth of the tested stored-product mites. α-AI at a concentration of 10 mg g(-1) exerted a growth-suppressive effect that depended on the diet and species of the mites. The growth rate of A. siro was affected by the type of diet and was higher on carbohydrate diet than on the protein diet, the suppressive effect of α-AI was on the both diets. The growth-suppressive effect of α-AI on L. destructor and T. putrescentiae was significant when they were fed the protein diet but not when they were fed the carbohydrate diet. The higher resistance of tested mites to α-AI (proteinaceous) compared to non-proteinaceous acarbose corresponds to a powerful proteotolytic system in the mite gut. The results are discussed in terms of the adaptability of mites to utilize the starch from food sources.

Bartonella-like bacteria carried by domestic mite species.

Bacteria of the genus Bartonella are carried by haematophagous mites, ticks, fleas and flies, and attack the erythrocytes of mammals. Here we describe a Bartonella-like clade, a distinct group related to Bartonellaceae, in stored-product mites (Acari: Astigmata) and a predatory mite Cheyletus eruditus (Acari: Prostigmata) based on the analysis of cloned 16S rRNA gene sequences. By using the clade-specific primers, closely related Bartonella-like 16S rRNA sequences were amplified from both laboratory colonies and field strains of three synanthropic mite species (Acarus siro, Lepidoglyphus destructor and Tyrophagus putrescentiae) and a predatory mite. Altogether, sequences of Bartonella-like bacteria were found in 11 strains, but were not detected in Dermatophagoides farinae and D. pteronyssinus and two strains of L. destructor. All obtained sequences formed a separate cluster branching as a sister group to Bartonellaceae and related to other separate clusters comprising uncultured bacterial clones from human skin and hemipteran insects (Nysius plebeius and Nysius sp.). The classification of sequences into operational taxonomic units (OTUs) showed a difference between A. siro and T. putrescentiae suggesting that the Bartonella-like bacteria are different in these two mite species. However, species specific sequences in separate OTUs were observed also for C. eruditus. Possible symbiotic interactions between Bartonella-like bacteria and their mite hosts are discussed.

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 effect of stored barley cultivars, temperature and humidity on population increase of Acarus siro, Lepidoglyphus destructor and Tyrophagus putrescentiae.

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.

Diet modulation of the microbiome of the pest storage mite Tyrophagus putrescentiae.

Storage mites colonize a wide spectrum of food commodities and adaptations to diets have been suggested as mechanisms enabling successful colonization. We characterized the response of seven unique Tyrophagus putrescentiae cultures (5K, 5L, 5N, 5P, 5Pi, 5S, and 5Tk) with different baseline microbiomes to different diets. The offered diets included a rearing diet, protein-enriched diet, oat flakes, and sunflower seeds. Microbiome characterization was performed using 16S ribosomal RNA (rRNA) gene amplicon sequencing and 16S rRNA gene quantitative PCR. The mite culture microbiomes were classified into four groups: (i) Sodalis-dominated (5Pi), (ii) Wolbachia-dominated (5N and 5P), (iii) Cardinium-dominated (5L and 5S), and (iv) asymbiontic (5K and 5Tk) mites dominated by Bacillus and Bartonella. Mite growth rates were most strongly affected by nutrients in the diet, while respiration and microbial community profiles were largely influenced by mite culture. While growth rate was not directly explained by microbiome composition, microbiomes strongly influenced mite fitness as measured by respiration. While diet significantly influenced microbial profiles in all cultures, the effect of diet differed in impact between cultures (5Pi > 5S > 5N > 5K > 5Tk > 5L > 5P). Furthermore, no new bacterial taxa were acquired by mites after dietary changes. Bacteria from the taxa Bacillus, Bartonella-like, Solitalea-like, Kocuria, and Sodalis-like contributed most strongly to differentiating mite-associated microbiomes.