Harnessing microfluidic streak plate technique to investigate the gut microbiome of Reticulitermes chinensis

Abstract

The termite gut microbiome is a model system to investigate microbial interactions and their associations with host. For decades, extensive research with molecular tools and conventional cultivation method has been carried out to define the microbial diversity in termite gut. Yet, many bacterial groups of the termite gut microbiome have not been successfully cultivated in laboratory. In this study, we adapted the recently developed microfluidic streak plate (MSP) technique for cultivation of termite gut microbial communities at both aerobic and anaerobic conditions. We found that 99 operational taxonomic units (OTUs) were cultivable by MSP approach and 18 OTUs were documented first time for termite gut microbiota. Further analysis of the bacterial diversities derived by culture-dependent MSP approach and culture-independent 16S rRNA gene typing revealed that both methods have bias in recovery of gut microbiota. In total 396 strains were isolated with MSP technique, and potential new taxa at species and/or genus levels were obtained that were phylogenetically related to Burkholderia, Micrococcus, and Dysgonomonas. Results from this study indicate that MSP technique is applicable for cultivating previously unknown and new microbial groups of termite gut microbiota.

Diversity of Bacterial Biota in Capnodis tenebrionis (Coleoptera: Buprestidae) Larvae

“…The Microvirga (formerly Balneimonas ) and Methylobacterium species are ubiquitous in nature, mainly soil and water, but also plants’ phylloplane [24,25,26]. Bacteria of the Microvirga and Methylobacterium genera have recently been found essential for efficient digestion of lignocellulose in the gut of the wood-feeding termite Reticulitermes chinensis (Isoptera: Rhinotermitidae) [27], that digest glucosyl and xylosyl residues from lignocellulose [28]. Methylobacterium spp.…”

Bacteria Cultivated From Sponges and Bacteria Not Yet Cultivated From Sponges—A Review

“…It is likely that fast-growing microbes compete for resources with uncultured slow-growing species. As such, it may be important to separate single microbial cells from complex microbial communities, e.g., through dilution-to-extinction, microfluidics, flow cytometry, micromanipulation, and compartmentalization to grow targeted bacteria physically separated from each other ( Ben-Dov et al, 2009 ; Ishii et al, 2010 ; Ma et al, 2014 ; Pivetal et al, 2014 ; Jiang et al, 2016 ; Zhou et al, 2018 ; Benítez et al, 2021 ), while perhaps retaining possibilities for interactions using the cultivation setups listed earlier in this paragraph. These methods have led to the isolation of previously uncultured bacterial species of clades SAR11 (Alphaproteobacteria), OM43 (Betaproteobacteria), SAR92 (Gammaproteobacteria), and OM60/OM241 (γ subclass) ( Connon and Giovannoni, 2002 ; Simu and Hagström, 2004 ; Stingl et al, 2007 ; Song et al, 2009 ) that are numerically abundant in their environment.…”

One cell at a time: droplet-based microbial cultivation, screening and sequencing

“…3e). Later, Zhou et al (2018) used the MSP platform under anaerobic conditions to separate several potentially new species and genera from the gut of Reticulitermes chinensis The MSP platform was further improved for long-term cultivation to tackle the problem of rare marine microbe isolation (Hu et al 2020). The improved MSP platform increased the recovery of slow-growing microbes and enabled longterm cultivation of up to five months.…”