Abstract

Chironomids are aquatic insects that undergo a complete metamorphosis of four life stages. Here we studied, for the first time, the microbiota composition of Chironomus circumdatus, a tropical midge species, both from the Mula and Mutha Rivers in Pune, India and as a laboratory-reared culture. We generated a comparative microbial profile of the eggs, larvae and pupae, the three aquatic life stages of C. circumdatus. Non-metric multidimensional scaling analysis (NMDS) demonstrated that the developmental stage had a more prominent effect on the microbiota composition compared to the sampling location. Notably, the microbiota composition of the egg masses from the different sampling points clustered together and differed from laboratory culture larvae. Proteobacteria was the dominant phylum in all the environmental and laboratory-reared egg masses and pupal samples, and in the laboratory-reared larvae, while Fusobacteria was the dominant phylum in the larvae collected from the field environment. The most abundant genera were Cetobacterium, Aeromonas, Dysgonomonas, Vibrio, and Flavobacterium. The ten amplicon sequence variants (ASVs) that most significantly contributed to differences in microbiota composition between the three sampled locations were: Burkholderiaceae (ASVs 04 and 37), C39 (Rhodocyclaceae, ASV 14), Vibrio (ASV 07), Arcobacter (ASV 21), Sphaerotilus (ASV 22), Bacteroidia (ASVs 12 and 28), Flavobacterium (ASV 29), and Gottschalkia (ASV 10). No significant differences were found in the microbial richness (Chao1) or diversity (Shannon H’) of the three sampled locations. In contrast, significant differences were found between the microbial richness of the three life stages. Studying the microbiota of this Chironomus species may contribute to a better understanding of the association of C. circumdatus and its microbial inhabitants.

Highlights

  • Chironomids (Diptera: Chironomidae), known as nonbiting midges, are aquatic insects that undergo a complete metamorphosis of four life stages

  • The microbiota compositions of the three life stages of C. circumdatus that were sampled from three environments were studied using Illumina sequencing of the 16S rRNA gene

  • To the best of our knowledge, this is the first study that investigates the changes in the bacterial composition at the different metamorphic life stages of C. circumdatus and compares them across different sampling locations

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Summary

Introduction

Chironomids (Diptera: Chironomidae), known as nonbiting midges, are aquatic insects that undergo a complete metamorphosis of four life stages (egg, larva, pupa, and adult). Females of the genus Chironomus deposit egg masses on biotic or abiotic objects that float on the surface of water bodies. The first larval instars feed on the egg mass remains and swim toward the bottom of the water body where they build a silken tube (Thorat and Nath, 2018). The larvae feed by extending their head and anterior part of their body outside the tube. These housing-cum-feeding tubes protect the larvae from their fish and invertebrate predators and other environmental stressors (Macchiusi and Baker, 1992; Thorat et al, 2017, 2020; Laviad-Shitrit et al, 2021).

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