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Bacterial Diversity in Solenopsis invicta and Solenopsis geminata Ant Colonies Characterized by 16S amplicon 454 Pyrosequencing

  • Environmental Microbiology
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Abstract

Social insects harbor diverse assemblages of bacterial microbes, which may play a crucial role in the success or failure of biological invasions. The invasive fire ant Solenopsis invicta (Formicidae, Hymenoptera) is a model system for understanding the dynamics of invasive social insects and their biological control. However, little is known about microbes as biotic factors influencing the success or failure of ant invasions. This pilot study is the first attempt to characterize and compare microbial communities associated with the introduced S. invicta and the native Solenopsis geminata in the USA. Using 16S amplicon 454 pyrosequencing, bacterial communities of workers, brood, and soil from nest walls were compared between neighboring S. invicta and S. geminata colonies at Brackenridge Field Laboratory, Austin, Texas, with the aim of identifying potential pathogenic, commensal, or mutualistic microbial associates. Two samples of S. geminata workers showed high counts of Spiroplasma bacteria, a known pathogen or mutualist of other insects. A subsequent analysis using PCR and sequencing confirmed the presence of Spiroplasma in additional colonies of both Solenopsis species. Wolbachia was found in one alate sample of S. geminata, while one brood sample of S. invicta had a high count of Lactococcus. As expected, ant samples from both species showed much lower microbial diversity than the surrounding soil. Both ant species had similar overall bacterial diversities, although little overlap in specific microbes. To properly characterize a single bacterial community associated with a Solenopsis ant sample, rarefaction analyses indicate that it is necessary to obtain 5,000–10,000 sequences. Overall, 16S amplicon 454 pyrosequencing appears to be a cost-effective approach to screen whole microbial diversity associated with invasive ant species.

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Acknowledgments

We thank Edward LeBrun for assisting in the collection of samples. Larry Gilbert, Edward LeBrun, and Jon Seal provided constructive comments on the manuscript. The work was funded by the Lee and Ramona Bass Foundation, the Helen C. Kleberg & Robert J. Kleberg Foundation, NSF Grant DEB-0639879, and by the W.M. Wheeler Lost-Pines endowment from the University of Texas at Austin.

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Authors and Affiliations

  1. Section of Integrative Biology, University of Texas at Austin, 2401 Speedway Drive C0930, Austin, TX, 78712, USA

    Heather D. Ishak, Ruchira Sen, Katrin Kellner, Eli Meyer & Ulrich G. Mueller

  2. Brackenridge Field Laboratory, University of Texas at Austin, 1 University Station, Austin, TX, 78712, USA

    Rob Plowes & Dora A. Estrada

  3. Medical Biofilm Research Institute, 4321 Marsha Sharp Freeway, Lubbock, TX, 79407, USA

    Scot E. Dowd

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  1. Heather D. Ishak
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Correspondence to Heather D. Ishak.

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Supplementary Figure 1

Rarefaction curves for each nest for using 1% (species level) and 3% (generic level) OTU designations (DOC 1312 kb)

Supplementary Figure 2

UniFrac weighted principal coordinate analyses (PCoA) for all S. geminata and S. invicta ant samples. The arrow indicates two S. geminata workers (A4 and A9) which have overlapping symbols due to their high numbers of Spiroplasma sequences. The bacterial communities between S. geminata and S. invicta have some overlap and are not distinctly separate (DOC 308 kb)

Supplementary Table 1

BLAST results according to its nearest genus (matched at 100–75% identity) for all 454-generated sequences and analyzed separately for each sample providing relative abundance percentages (XLS 222 kb)

Supplementary Table 2

Conservative BLAST results for all 454 generated sequence data (XLS 416 kb)

Supplementary Table 3

Unique sequence ID’s, BLAST hit, and identity scores for all 454-generated sequences (XLS 9146 kb)

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Ishak, H.D., Plowes, R., Sen, R. et al. Bacterial Diversity in Solenopsis invicta and Solenopsis geminata Ant Colonies Characterized by 16S amplicon 454 Pyrosequencing. Microb Ecol 61, 821–831 (2011). https://doi.org/10.1007/s00248-010-9793-4

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