Abstract

Simple SummaryLike most insects, those that feed on both prey and plant materials harbor symbiotic bacteria in their body. Yet the involvement of bacteria in the feeding habits of these omnivorous consumers has yet to be investigated. In the present study, we took the first step toward testing the hypothesis that bacterial symbionts are involved in the feeding habits of the omnivorous bug Macrolophus pygmaeus. We (I) characterized the microbiome (the assembly of bacteria and fungi) of M. pygmaeus, and (II) determined the identity and location of the most dominant bacteria species within the host body. We found that M. pygmaeus microbiome is dominated by two Rickettsia species, R. belli and R. limoniae. These bacteria are found in high numbers in the digestive system of the bug, each exhibiting a unique distribution pattern, and for the most part, do not share the same cells in the gut. These results strongly suggest that the host bug may gain some nutritional benefits by hosting the two dominant symbiotic bacteria in its gut.Bacterial symbionts in arthropods are common, vary in their effects, and can dramatically influence the outcome of biological control efforts. Macrolophus pygmaeus (Heteroptera: Miridae), a key component of biological control programs, is mainly predaceous but may also display phytophagy. M. pygmaeus hosts symbiotic Wolbachia, which induce cytoplasmic incompatibility, and two Rickettsia species, R. bellii and R. limoniae, which are found in all individuals tested. To test possible involvement of the two Rickettsia species in the feeding habits of M. pygmaeus, we first showed that the microbiome of the insect is dominated by these three symbionts, and later described the distribution pattern of the two Rickettsia species in its digestive system. Although both Rickettsia species were located in certain gut bacteriocyes, in caeca and in Malpighian tubules of both sexes, each species has a unique cellular occupancy pattern and specific distribution along digestive system compartments. Infrequently, both species were found in a cell. In females, both Rickettsia species were detected in the germarium, the apical end of the ovarioles within the ovaries, but not in oocytes. Although the cause for these Rickettsia distribution patterns is yet unknown, it is likely linked to host nutrition while feeding on prey or plants.

Highlights

  • Colonization by microorganisms is a universal phenomenon among animals including insects, one of the most successful groups in the animal kingdom [1]

  • The bacterial data set originating from five samples, resulted in a total of 94,293 reads mapped to 28 amplicon sequence variantsusing (ASVs) which were further analyzed (Table S1)

  • ASVs were prevalent in all five samples and included different Actinobacteria

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Summary

Introduction

Colonization by microorganisms is a universal phenomenon among animals including insects, one of the most successful groups in the animal kingdom [1]. The omnivorous bug N. tenuis, for example, is known to harbor both Rickettsia and Wolbachia endosymbionts, and both have been found in host ovaries, but in the gut tissue as well [20] Bacteria of these same genera have been reported by Machtelinckx et al [21] in M. pygmaeus, a key natural enemy of various economically important agricultural pests of greenhouse vegetable crops [17]. The overall objective of our ongoing research is to test the possible involvement of bacterial symbionts in the omnivorous feeding habits of M. pygmaeus. To this end, we (I) characterized the microbiome of Insects 2020, 11, x FOR PEER REVIEW becomes scarce [22].

Insect
Microbiota
Screening for Rickettsia bellii and Rickettsia limoniae Prevalance
Morphology of the Digestive System
PCR for Ovaries and Digestive System
Microbiota Characterization
Morphology of Digestive System
Fluorescence in Situ Hybridization
Discussion
Conclusions

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