Microbiome analysis reveals the abundance of bacterial pathogens in Rousettus leschenaultii guano - PubMed (original) (raw)

Microbiome analysis reveals the abundance of bacterial pathogens in Rousettus leschenaultii guano

Sunil Banskar et al. Sci Rep. 2016.

Abstract

Bats are crucial for proper functioning of an ecosystem. They provide various important services to ecosystem and environment. While, bats are well-known carrier of pathogenic viruses, their possible role as a potential carrier of pathogenic bacteria is under-explored. Here, using culture-based approach, employing multiple bacteriological media, over thousand bacteria were cultivated and identified from Rousettus leschenaultii (a frugivorous bat species), the majority of which were from the family Enterobacteriaceae and putative pathogens. Next, pathogenic potential of most frequently cultivated component of microbiome i.e. Escherichia coli was assessed to identify its known pathotypes which revealed the presence of virulent factors in many cultivated E. coli isolates. Applying in-depth bacterial community analysis using high-throughput 16 S rRNA gene sequencing, a high inter-individual variation was observed among the studied guano samples. Interestingly, a higher diversity of bacterial communities was observed in decaying guano representative. The search against human pathogenic bacteria database at 97% identity, a small proportion of sequences were found associated to well-known human pathogens. The present study thus indicates that this bat species may carry potential bacterial pathogens and advice to study the effect of these pathogens on bats itself and the probable mode of transmission to humans and other animals.

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Figures

Figure 1

Figure 1. Venn diagram showing the distribution of bacterial species obtained from different bacteriological media.

LA; Luria Hiveg Agar, STR Streptomycetes Isolation Agar, ZMA; Zobells Marine Agar, AK; Arret & Kirshbaum medium.

Figure 2

Figure 2. Genus level distribution of bacterial isolates obtained from fresh bat guano.

Figure 3

Figure 3. Escherichia coli isolates of R. leschenaultii guano carrying different virulent genes belonging to different pathotypes.

None of the isolates were positive for ipaH, sta and stx1 gene. E. coli isolates G8ZM15 and G28BLA40 carried stb and stx2 genes. (hlyA = gene for alpha hemolysin, ibeA = Invasion of brain endothelium (extraintestinal E. coli (ExPEC)); east1 = heat-stable enterotoxin (entero-aggregative E. coli (EaggEC)); stx1 = Shiga toxin-1, stx2 = Shiga toxin-2 for enterohemorrhagic E. coli (EHEC) identification; ipah = Invasion Plasmid antigen enteroinvasive E. coli (EIEC); stb = Heat stable toxin-b, sta = Heat stable toxin-a enterotoxigenic E. coli (ETEC)).

Figure 4

Figure 4. Phylum level distribution of Ion Torrent sequences of different guano samples.

CGS = Cave Ground Surface Soil; G = Guano.

Figure 5

Figure 5. Inter-sample variation in gut microbiome (calculated from bacterial phyla constituting core microbiome).

Different color represents the different samples. Different shapes indicate the different phylum of bacteria. The error bars indicate the standard deviation of mean of relative proportion of Ion Torrent sequences.

Figure 6

Figure 6. Phylum level distribution of different microbiome studies compared.

Figure 7

Figure 7. Comparison of composite guano and other samples used in this study.

Both representatives of decaying guano (Red and Blue) are far apart from the rest of fresh bat guano samples (green) in PCoA plot generated using the Jaccard distance.

Figure 8

Figure 8. Heat map showing the relative abundance of Ion Torrent sequences identified as bacterial pathogen at 97% sequence identity cutoff.

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