The effect of sodium hypochlorite on the growth of <i>Batrachochytrium dendrobatidis</i> (Bd).

<p>Change in optical density (OD) of Bd four days after exposure to various concentrations of sodium hypochlorite for (A) 0.5 min, (B) 1 min, (C) 5 min, and (D) 15 min. Error bars represent standard error.</p

Minimum lethal concentration of sodium hypochlorite for the amphibian pathogen <i>Batrachochytrium dendrobatidis</i>

<div><p>Sodium hypochlorite (NaOCl) is the active ingredient in household bleach and is commonly used as a disinfectant to clean equipment contaminated by the amphibian pathogen <i>Batrachochytrium dendrobatidis</i> (Bd) in lab husbandry and field studies. We conducted a series of replicated exposure trials using a single Global Pandemic Lineage Bd isolate from Panama (JEL 310) and concentrations of NaOCl ranging from 0.006% to 0.6% for exposure times ranging from 30 seconds to 15 minutes to determine the minimum lethal concentration of NaOCl for this isolate of Bd. Sodium hypochlorite completely killed Bd at a concentration of 0.03% during a 15-minute exposure time, while 0.12% NaOCl was effective at all exposure times (30s-15min).</p></div

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

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

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

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

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

Hellbender skin Bd transcriptome metadata

In this study, we explored the efficacy of Bd vaccination on hellbender salamanders and functionally characterized host, host-associated microbiome, and pathogen throughout immunization and infection. We compared the function of hellbender skin, the skin microbiome, and Bd with a metatranscriptome approach at four experimental time points, (i) pre-vaccination and pre-infection, (ii) post-vaccination and pre-infection, (iii) early infection, and (iv) late infection.</p

Hellbender skin prokaryote transcriptome BLAST xml

In this study, we explored the efficacy of Bd vaccination on hellbender salamanders and functionally characterized host, host-associated microbiome, and pathogen throughout immunization and infection. We compared the function of hellbender skin, the skin microbiome, and Bd with a metatranscriptome approach at four experimental time points, (i) pre-vaccination and pre-infection, (ii) post-vaccination and pre-infection, (iii) early infection, and (iv) late infection.</p

Hellbender skin transcriptome

In this study, we explored the efficacy of Bd vaccination on hellbender salamanders and functionally characterized host, host-associated microbiome, and pathogen throughout immunization and infection. We compared the function of hellbender skin, the skin microbiome, and Bd with a metatranscriptome approach at four experimental time points, (i) pre-vaccination and pre-infection, (ii) post-vaccination and pre-infection, (iii) early infection, and (iv) late infection.</p