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.2020 Oct 13;10(10):406.
doi: 10.3390/metabo10100406.

Metabolites from Microbes Isolated from the Skin of the Panamanian Rocket FrogColostethus panamansis (Anura: Dendrobatidae)

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Metabolites from Microbes Isolated from the Skin of the Panamanian Rocket FrogColostethus panamansis (Anura: Dendrobatidae)

Christian Martin H et al. Metabolites..

Abstract

The Panamanian rocket frogColostethus panamansis (family Dendrobatidae) has been affected by chytridiomycosis, a deadly disease caused by the fungusBatrachochytrium dendrobatidis (Bd). While there are still uninfected frogs, we set out to isolate microbes from anatomically distinct regions in an effort to create a cultivable resource within Panama for potential drug/agricultural/ecological applications that perhaps could also be used as part of a strategy to protect frogs from infections. To understand if there are specific anatomies that should be explored in future applications of this resource, we mapped skin-associated bacteria ofC. panamansis and their metabolite production potential by mass spectrometry on a 3D model. Our results indicate that five bacterial families (Enterobacteriaceae, Comamonadaceae, Aeromonadaceae, Staphylococcaceae and Pseudomonadaceae) dominate the cultivable microbes from the skin ofC. panamansis. The combination of microbial classification and molecular analysis in relation to the anti-Bd inhibitory databases reveals the resource has future potential for amphibian conservation.

Keywords: 16S rRNA sequencing; 3D molecular cartography; Colostethus panamansis; Dendrobatidae; feature-based molecular networking; skin-associated bacteria.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Frequency of isolated bacteria from the skin surface ofC. panamansis. (A) Relative abundances of bacterial families from the skin ofC. panamansis based on 16S rRNA amplicon sequencing. (B) The proportional abundance (as a percent of total sequences) of bacterial families based on body parts. (C) 3D topographic map ofC. panamansis based on the five most abundant bacterial families isolated at each body location. Red represents the highest percentage of each family that was isolated and blue the lowest percentage.
Figure 2
Figure 2
Molecular network of specialized metabolites (<300 Daltons) annotated in crude extracts of bacterial isolates from skin ofC. panamansis by LC-MS/MS analysis. Highlighted nodes correspond to annotated metabolites by GNPS (yellow) and SIRIUS/CSIFingerID (purple). Pie charts inside nodes denotes abundance while colors correspond to the bacterial family where the feature was found. Edge width between nodes displays cosine score values, which are related to the spectral similarities between nodes.
Figure 3
Figure 3
Molecular network of annotated peptides identified in crude extracts of bacterial isolates from skin ofC. panamansis by LC-MS/MS analysis. Highlighted nodes correspond to annotated metabolites by GNPS (yellow) and SIRIUS CSIFingerID (purple). Pie charts inside nodes denotes abundance while colors correspond to the bacterial family where the feature was found. Edge width between nodes displays cosine score values, which are related to the spectral similarities between nodes.
Figure 4
Figure 4
Molecular cartography of annotated metabolites identified from bacterial isolates sampled from the skin surface ofC. panamansis. These metabolites were annotated as tryptophan derivatives (1,2), phenylalanine (3), 3-indoleacetic acid (4), diketopiperazines (5–10) and peptides (11–18). Intensities are related to the abundance of the specialized metabolites in the samples at each body part. Red represents the highest abundance while blue represents the lowest abundance.
Figure 5
Figure 5
Phylogenetic tree and structures of annotated metabolites from 170 cultivable bacteria isolated from the skin ofC. panamansis and a heatmap that displays the abundance of specialized metabolites annotated as molecular features by feature based molecular networking-analysis.
Figure 6
Figure 6
Relative abundances of 791 bacterial sequences fromC. panamansis and anti-Bd published sequences. (A) Relative abundance of 16S rRNA sequences fromC. panamansis and anti-Bd sequences from published databases based on their bacterial family. (B) Comparison between the five most abundant bacterial families inC. panamansis andBd-inhibitory sequences (light green) and the distribution of isolates based on the coronal plane inC. panamansis is also displayed.
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