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Microbiota Analysis Using Two-step PCR and Next-generation 16S rRNA Gene Sequencing
Published on: October 15, 2019
Microbial Communities of Stored Product Mites: Variation by Species and Population
Jan Hubert1,2, Marta Nesvorna3, Stefan J Green4
1Crop Research Institute, Drnovska 507/73, CZ-161 06, Prague 6-Ruzyne, Czechia. hubert@vurv.cz.
Abstract:
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.
Insights
Stored product mites harbor complex microbiomes, with variations within species exceeding those between species. These microbial communities significantly impact mite fitness and population growth.
Area of Science:
- Microbiology
- Ecology
- Entomology
Background:
- Arthropod-associated microbes influence host fitness, pathogen resistance, and disease transmission.
- Stored product mites contaminate food with allergens and microorganisms, impacting food safety and quality.
Purpose of the Study:
- To investigate the microbial structure and abundance in stored product mites and their environment.
- To determine the impact of mite-associated microbes on mite fitness and population dynamics.
Main Methods:
- Quantitative PCR and 16S ribosomal RNA (rRNA) gene amplicon sequencing were used.
- Microbial communities were analyzed in mite diets, eggs, feces, and bodies from 16 populations of three mite species.
- Bacterial 16S rRNA gene copy numbers were quantified to assess absolute bacterial abundance.
Main Results:
- Mite microbiomes were complex, dominated by intracellular symbionts (Cardinium, Wolbachia), gut symbionts (Solitalea, Bartonella, Sodalis), and environmental bacteria (Bacillus, Staphylococcus, Kocuria).
- Microbial community differences were greater between conspecific mite populations than between different mite species.
- Mite fitness correlated positively with bacteria in the spent growth medium (SPGM) and negatively with bacteria in mite bodies.
Conclusions:
- Host-microbe interactions are critical throughout mite development, influencing their environmental microbiome.
- Mite-associated microorganisms play a key role in modulating mite pest properties, including population growth.
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