Open AccessAn investigation of conventional microbial culture for the Naja atra bite wound, and the comparison between culture-based 16S Sanger sequencing and 16S metagenomics of the snake oropharyngeal bacterial microbiota
Author(s) -
Yan–Chiao Mao,
HanNi Chuang,
Chien-Hung Shih,
Han-Hsueh Hsieh,
Yuhan Jiang,
Liao-Chun Chiang,
WenLoung Lin,
TzuHung Hsiao,
PoYu Liu
Publication year - 2021
Publication title -
plos neglected tropical diseases
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.99
H-Index - 135
eISSN - 1935-2735
pISSN - 1935-2727
DOI - 10.1371/journal.pntd.0009331
Subject(s) - metagenomics , biology , microbiology and biotechnology , proteus mirabilis , stenotrophomonas maltophilia , sanger sequencing , microbiological culture , pseudomonas aeruginosa , bacteria , dna sequencing , genetics , gene
Naja atra is a major venomous snake found in Taiwan. The bite of this snake causes extensive wound necrosis or necrotizing soft tissue infection. Conventional microbial culture-based techniques may fail to identify potential human pathogens and render antibiotics ineffective in the management of wound infection. Therefore, we evaluated 16S Sanger sequencing and next-generation sequencing (NGS) to identify bacterial species in the oropharynx of N . atra . Using conventional microbial culture methods and the VITEK 2 system, we isolated nine species from snakebite wounds. On the basis of the 16S Sanger sequencing of bacterial clones from agar plates, we identified 18 bacterial species in the oropharynx of N . atra , including Morganella morganii , Proteus vulgaris , and Proteus mirabilis , which were also present in the infected bite wound. Using NGS of 16S metagenomics, we uncovered more than 286 bacterial species in the oropharynx of N . atra . In addition, the bacterial species identified using 16S Sanger sequencing accounted for only 2% of those identified through NGS of 16S metagenomics. The bacterial microbiota of the oropharynx of N . atra were modeled better using NGS of 16S metagenomics compared to microbial culture-based techniques. Stenotrophomonas maltophilia , Acinetobacter baumannii , and Proteus penneri were also identified in the NGS of 16S metagenomics. Understanding the bacterial microbiota that are native to the oropharynx of N . atra , in addition to the bite wound, may have additional therapeutic implications regarding empiric antibiotic selection for managing N . atra bites.