Differences in the number of OTUs among animal diets were evaluated using an ANOVA (see Tables in manuscript and supplementary information). Here, each dietary treatment was analyzed separately. For multivariate analysis, the 16S OTUs distances among samples first were calculated using the unweighted (bacterial counts as 0 and 1 observations) UniFrac

distance measure ([20], which measures the phylogenetic distances among samples. The weighted (actual abundance) UniFrac distance measure was used because it also considers the relative abundance of each OTU (16S rRNA read) when calculating phylogenetic distances. Principle coordinates analysis (PCoA) was used Erismodegib datasheet to display these differences in 2 dimensions, thereby facilitating an overall assessment of variability in the entire microbiome among samples. To test for multivariate differences among treatment groups, distance based redundancy analysis (dbRDA) [21] was used. find more In addition, the relative abundances of all genera were evaluated using an ANOVA. Here, relative abundances were transformed (p’ = arcsine (√p)) before analysis, and analyses

were conducted separately for each of the diets. As an initial screening evaluation, uncontrolled p-values were used to screen taxa. Data are illustrated in figures in the manuscript and supplementary information. Rarefaction curves and UniFrac distances were calculated using QIIME [22], and all other analyses were conducted in R [23], using the vegan [24] and labdsv [25] packages. Double hierarchal cluster analysis was conducted using NCSS 2007 Reverse transcriptase software (NCSS, Kaysville, UT) and one-way ANOVA was also conducted using JMP9 software (JMP, SAS, Cary, NC). Acknowledgements The authors recognize

Lana Castleberry for the preparation of community DNA samples for analysis. Electronic supplementary material Additional file 1: Figure S1. Evaluation of Bacteroidetes and Firmicutes relative abundance to the influence of dietary treatments, (A) One-way Analysis of Firmicutes by Treatment, (B) One-way Analysis of Bacteroidetes by Treatment, and (C) Matched pair comparisons testing the response of the ratio of abundances observed between Bacteroidetes and Firmicutes revealing no significant difference between and amongst treatments. (PPT 692 KB) Additional file 2: Figure S2. Evaluation of Phyla showing a response (significant < 0.05, or influenced < 0.1) to dietary treatments (A) Oneway analysis of Synergistetes by treatment, (B) Oneway analysis of WS3 by treatment, (C) Oneway analysis of Actinobacteria by treatment, (D) Oneway analysis of Spirochaetes by treatment. (PPT 110 KB) Additional file 3: Figure S3. Effect of wet DG’s on Beef Cattle Fecal Microbiota.