Abstract

Purpose: To employ next-generation sequencing (NGS) to investigate the diversity of microorganisms and pathogenic bacteria from a water source in Tai Lake, China, in winter. Methods: Water samples from the same source were collected over a period of 3 months (December 2013 to February 2014), and their physicochemical characteristics were determined. The DNA of the samples were extracted and amplified by polymerase chain reaction (PCR). The PCR products were sequenced by Miseq PE300 pyro sequencing platform. The results for 16S rDNA were analysed using visualization software Gephi, and the 16S rDNA gene pool of known pathogenic bacteria was established. Results: A total of 144,292 16S rDNA gene sequences were obtained and ranked by RDP classifier. The average length of the sequences was 395.66 bp. They revealed 580 operational taxonomic units (OTUs) classified into 16 phyla. A full length of 16S rDNA gene database of common pathogenic bacteria was established. After blasting, 17 species of pathogenic bacteria were found. The most abundant potential human pathogenic bacteria were affiliated to B. tribocorum . Most environmental factors had significant impact on pathogenic bacteria. Conclusion: These results indicate that NGS can be used for the simultaneous detection of most recognized water-borne pathogenic bacteria. Variations in microorganisms in water source at different periods in winter can provide insight into the diversity of microorganisms in the water. Keywords: Next-generation sequencing, Pathogenic bacteria, Diversity, 16S rDNA gene

Highlights

  • Water-borne diseases are illnesses caused by intake of water that harbors pathogenic microorganisms [1]

  • After highthroughput pyro sequencing and sequence diversity analysis, a csv file was built, which could be received by Gephi base on the genera, environment and abundance

  • Comprehensive the 16S rDNA sequences of bacteria and pathogens in water samples from Tai lake in winter, through next-generation sequencing (NGS) indicate that the pathogenic bacterial species were relatively stable in the lake in winter, suggesting a potential microbial risk in the water source

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Summary

Introduction

Water-borne diseases are illnesses caused by intake of water that harbors pathogenic microorganisms [1]. Water-borne diseases are caused by water-borne pathogens, which are recognized as the microbial risk in drinking water [3]. In addition to pathogens, drinking water may harbor various animal and plant pathogenic microorganisms [4], which survive even after water processing [5]. These microorganisms persist in drinking water, thereby posing danger to people’s health. Pure culture method has been used to detect pathogenic microorganisms. It is necessary to explore novel methods for detecting pathogenic microorganisms in drinking water

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