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      Localization of Bacterial Communities within Gut Compartments across Cephalotes Turtle Ants

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          Abstract

          Gut compartments play an important role in structuring the microbial community within individual ants. The gut chambers of the turtle ant digestive tract differ remarkably in symbiont abundance and diversity.

          ABSTRACT

          Microbial communities within the animal digestive tract often provide important functions for their hosts. The composition of eukaryotes’ gut bacteria can be shaped by host diet, vertical bacterial transmission, and physiological variation within the digestive tract. In several ant taxa, recent findings have demonstrated that nitrogen provisioning by symbiotic bacteria makes up for deficiencies in herbivorous diets. Using 16S rRNA amplicon sequencing and quantitative PCR, this study examined bacterial communities at a fine scale across one such animal group, the turtle ant genus Cephalotes. We analyzed the composition and colonization density across four portions of the digestive tract to understand how bacterial diversity is structured across gut compartments, potentially allowing for specific metabolic functions of benefit to the host. In addition, we aimed to understand if caste differentiation or host relatedness influences the gut bacterial communities of Cephalotes ants. Microbial communities were found to vary strongly across Cephalotes gut compartments in ways that transcend both caste and host phylogeny. Despite this, caste and host phylogeny still have detectable effects. We demonstrated microbial community divergence across gut compartments, possibly due to the various function of each gut compartment for digestion.

          IMPORTANCE Gut compartments play an important role in structuring the microbial community within individual ants. The gut chambers of the turtle ant digestive tract differ remarkably in symbiont abundance and diversity. Furthermore, caste type explains some variation in the microbiome composition. Finally, the evolutionary history of the Cephalotes species structures the microbiome in our study, which elucidates a trend in which related ants maintain related microbiomes, conceivably owing to cospeciation. Amazingly, gut compartment-specific signatures of microbial diversity, relative abundance, composition, and abundance have been conserved over Cephalotes evolutionary history, signifying that this symbiosis has been largely stable for over 50 million years.

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          DADA2: High resolution sample inference from Illumina amplicon data

          We present DADA2, a software package that models and corrects Illumina-sequenced amplicon errors. DADA2 infers sample sequences exactly, without coarse-graining into OTUs, and resolves differences of as little as one nucleotide. In several mock communities DADA2 identified more real variants and output fewer spurious sequences than other methods. We applied DADA2 to vaginal samples from a cohort of pregnant women, revealing a diversity of previously undetected Lactobacillus crispatus variants.
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            RAxML version 8: a tool for phylogenetic analysis and post-analysis of large phylogenies

            Motivation: Phylogenies are increasingly used in all fields of medical and biological research. Moreover, because of the next-generation sequencing revolution, datasets used for conducting phylogenetic analyses grow at an unprecedented pace. RAxML (Randomized Axelerated Maximum Likelihood) is a popular program for phylogenetic analyses of large datasets under maximum likelihood. Since the last RAxML paper in 2006, it has been continuously maintained and extended to accommodate the increasingly growing input datasets and to serve the needs of the user community. Results: I present some of the most notable new features and extensions of RAxML, such as a substantial extension of substitution models and supported data types, the introduction of SSE3, AVX and AVX2 vector intrinsics, techniques for reducing the memory requirements of the code and a plethora of operations for conducting post-analyses on sets of trees. In addition, an up-to-date 50-page user manual covering all new RAxML options is available. Availability and implementation: The code is available under GNU GPL at https://github.com/stamatak/standard-RAxML. Contact: alexandros.stamatakis@h-its.org Supplementary information: Supplementary data are available at Bioinformatics online.
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              The SILVA ribosomal RNA gene database project: improved data processing and web-based tools

              SILVA (from Latin silva, forest, http://www.arb-silva.de) is a comprehensive web resource for up to date, quality-controlled databases of aligned ribosomal RNA (rRNA) gene sequences from the Bacteria, Archaea and Eukaryota domains and supplementary online services. The referred database release 111 (July 2012) contains 3 194 778 small subunit and 288 717 large subunit rRNA gene sequences. Since the initial description of the project, substantial new features have been introduced, including advanced quality control procedures, an improved rRNA gene aligner, online tools for probe and primer evaluation and optimized browsing, searching and downloading on the website. Furthermore, the extensively curated SILVA taxonomy and the new non-redundant SILVA datasets provide an ideal reference for high-throughput classification of data from next-generation sequencing approaches.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                Appl Environ Microbiol
                Appl Environ Microbiol
                aem
                aem
                AEM
                Applied and Environmental Microbiology
                American Society for Microbiology (1752 N St., N.W., Washington, DC )
                0099-2240
                1098-5336
                12 February 2021
                26 March 2021
                April 2021
                : 87
                : 8
                : e02803-20
                Affiliations
                [a ]University of Chicago, Committee on Evolutionary Biology, Chicago, Illinois, USA
                [b ]Drexel University, Department of Biodiversity, Earth and Environmental Science, Philadelphia, Pennsylvania, USA
                [c ]Cornell University, Department of Ecology and Evolutionary Biology, Ithaca, New York, USA
                [d ]Cornell University, Department of Entomology, Ithaca, New York, USA
                Chinese Academy of Sciences
                Author notes
                Address correspondence to Peter J. Flynn, pflynn@ 123456uchicago.edu .

                Citation Flynn PJ, D’Amelio CL, Sanders JG, Russell JA, Moreau CS. 2021. Localization of bacterial communities within gut compartments across Cephalotes turtle ants. Appl Environ Microbiol 87:e02803-20. https://doi.org/10.1128/AEM.02803-20.

                Author information
                https://orcid.org/0000-0002-3182-7671
                https://orcid.org/0000-0003-1139-5792
                Article
                02803-20
                10.1128/AEM.02803-20
                8091110
                33579688
                e03aa786-73d4-4a0f-aa4a-e80e9904283f
                Copyright © 2021 Flynn et al.

                This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

                History
                : 18 November 2020
                : 5 February 2021
                Page count
                Figures: 7, Tables: 3, Equations: 0, References: 89, Pages: 18, Words: 11449
                Funding
                Funded by: National Science Foundation (NSF), https://doi.org/10.13039/100000001;
                Award ID: 1900357
                Award Recipient : Award Recipient : Award Recipient : Award Recipient : Award Recipient :
                Categories
                Invertebrate Microbiology
                Custom metadata
                April 2021

                Microbiology & Virology
                gut microbes,turtle ants,localization,host-microbe,symbionts
                Microbiology & Virology
                gut microbes, turtle ants, localization, host-microbe, symbionts

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