Skin microbial communities of <i>Colostethus panamansis</i>.

<p>Communities before (circles, field samples) and after (crosses, day 48) treatments visualized by non-metric multidimensional scaling (nMDS) of T-RFLP analysis using enzymes HaeIII and MspI. Treatments are numbered to indicate probiotic bacterium <i>Lysinibacillus fusiformis</i> (1,2), control (3,4), and skin-wash transplant from the disease-resistant glass frog <i>Espadarana prosoblepon</i> (5,6). Microbial communities were not significantly different among treatments within each time-point represented by convex hulls. Distance between objects on the plot represents relative dissimilarity (axes are in arbitrary units). Stress <0.1 indicates strong community differences and stress >0.2 indicates that differences should be interpreted with caution. Statistical analyses are presented in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087101#pone-0087101-t001" target="_blank">Table 1</a>.</p

Representative chromatograms of skin defense peptides examined by HPLC-MS.

<p>(a) <i>Colostethus panamansis.</i> (b) <i>Espadarana prosoblepon</i>. Values of molecular weight and mean area for the detected peptides are reported in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0087101#pone-0087101-t002" target="_blank">Table 2</a>.</p

Peptide dry mass predicts a significant proportion of variation in peptide intensity determined by LC-MS.

<p>Overall, there was a significant correlation between peptide dry mass and rank abundance of peptide mass 1064. This relationship differed among treatments indicating a change in the relative abundance of the peptide components of skin secretions depending on microbial treatment. Transplant treated frogs had the highest relative abundance of peptide mass 1064 (lowest rank), and probiotic treated frogs the lowest relative abundance with controls intermediate. Probiotic treated frogs had significantly higher total quantity of peptides than transplant treated frogs (see text).</p

Statistical analysis of treatment differences in microbial communities described by T-RFLP using either Hae3 or Msp1 enzymes.

<p>Analysis of similarity (ANOSIM) and non-parametric multivariate analysis of variance (NPMANOVA) results are shown of the three <i>C. panamansis</i> treatments: P = probiotic treatment, C = control treatment, T = transplant treatment. Significant values identified by ANOSIM and NPMANOVA are indicated in bold.</p

Retention time (Rt), molecular weight (MW), prevalence, and mean relative area of each peptide based on HPLC-MS chromatograms.

<p>Area is relative to a consistently observed peak, MW 1064.0 for <i>C. panamansis</i> and MW 2681.2 for <i>E. prosoblepon</i>.</p