Conclusions Insect-associated microbiota can be difficult to classify using existing

databases [15]; The lack of cultured isolates or characterized species from insect environments and also the enormous diversity of hosts for the microbial communities is problematic. For example, when predefined, publically available datasets are used to train the RDP-NBC and classify sequences from the honey bee gut, an environment for which there are no cultured representatives, taxonomic classifications are unstable and inconsistent (Figure 2A). In contrast, the HBDB custom training sets effectively and confidently classify the bacteria in the honey bee gut. Results from our classification are consistent with previous studies of the honey bee gut using 16S rRNA clone libraries [17, 18], suggesting that the inclusion

of environment-specific, high-quality, BIBW2992 cell line full-length sequences in the training set can dramatically affect the classification results produced by the RDP-NBC. In addition, the larger, more diverse training sets (SILVA + bees and GG + bees), provided more stable and precise classifications, echoing results of previous studies and suggesting that breadth and depth in the RDP-NBC training set is crucial for more confident taxonomic classifications [11]. This result echoes those of other groups who have found that representation in training sets markedly affects RDP-NBC ACY-1215 purchase performance [11, 29]. Acknowledgements This work was funded by startup funds provided by Indiana University to ILGN. The manuscript benefited from the

critiques of four anonymous reviewers, to which we are thankful. Electronic supplementary material Additional file 1: Table S1. Total number of operational taxonomic units (97% ID) in either genetically uniform or genetically diverse colonies and classified as one of Mannose-binding protein-associated serine protease the honey bee specific taxonomic groups. (DOCX 48 KB) Additional file 2: Table S2. Top scoring blastn hits between full-length, bee specific sequences and the Greengenes training set. (XLSX 46 KB) Additional file 3: Figure S1. Phylogenetic placement of representative short read classified as Orbus by the RDP + bees training set. (DOCX 271 KB) References 1. Andersson AF, Lindberg M, Jakobsson H, Backhed F, Nyren P, Engstrand L: Comparative Analysis of Human Gut Microbiota by Barcoded Pyrosequencing. PLoS One 2008,3(7):e2836.PubMedCrossRef 2. Bates ST, Berg-Lyons D, Caporaso JG, Walters WA, Knight R, Fierer N: Examining the global Selleck VE822 distribution of dominant archaeal populations in soil. ISME J 2011,5(5):908–917.PubMedCrossRef 3. Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Lozupone CA, Turnbaugh PJ, Fierer N, Knight R: Global patterns of 16S rRNA diversity at a depth of millions of sequences per sample. P Natl Acad Sci USA 2011, 108:4516–4522.CrossRef 4.