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Surveying the microbiome of ants: Comparing 454 pyrosequencing with traditional methods to uncover bacterial diversity.

Field Museum of Natural History, Department of Zoology, 1400 S. Lake Shore Dr., Chicago, IL 60605, USA.
Applied and Environmental Microbiology (Impact Factor: 3.95). 11/2012; DOI: 10.1128/AEM.03107-12
Source: PubMed

ABSTRACT We are only beginning to understand the depth and breadth of microbial associations across the eukaryotic tree of life. Reliably assessing bacterial diversity is a key challenge and next-generation sequencing approaches are facilitating this endeavor. Here, we used 16S rRNA amplicon pyrosequencing to survey microbial diversity in ants. We compared 454 libraries with those from Sanger sequenced clone libraries as well as cultivation of live bacteria. Pyrosequencing yielded 95,656 bacterial 16S rRNA reads from 19 samples derived from four colonies of one ant species. The most dominant bacterial orders in the microbiome of the turtle ant Cephalotes varians were Rhizobiales, Burkholderiales, Opitutales, Xanthomonadales, and Campylobacterales as revealed through both 454 sequencing and cloning. Even after stringent quality filtering, pyrosequencing recovered 445 microbe OTUs not detected with traditional techniques. In comparing bacterial communities associated with specific tissues, we found that gut tissues had significantly higher diversity than non-gut tissues and many of the OTUs identified from these groups clustered within ant specific lineages indicating a deep coevolutionary history of Cephalotes ants and their associated microbes. These lineages likely function as nutritional symbionts. One of four ant colonies investigated was infected with a Spiroplasma sp. (order Entomoplasmatales), a potential ant pathogen. Our work shows that the microbiome associated with Cephalotes varians is dominated by a few dozen bacterial lineages, and that 454 sequencing is a cost efficient tool to screen ant symbiont diversity.

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