16S Next-generation sequencing and quantitative PCR reveal the distribution of potential pathogens in the Liaohe Estuary

Abstract

The existence of potentially pathogenic bacteria seriously threatens aquatic animals and human health. Estuaries are closely related to human activities, and the detection of pathogens is important for aquaculture and public health. However, monitoring only indicator microorganisms and pathogens is not enough to accurately and comprehensively estimate water pollution. Here, the diversity of potentially pathogenic bacteria in water samples from the Liaohe estuary was profiled using 16S next-generation sequencing (16S NGS) and quantitative polymerase chain reaction (qPCR) analysis. The results showed that the dominant genera of environmental pathogens were Pseudomonas, Vibrio, Mycobacterium, Acinetobacter, Exiguobacterium, Sphingomonas, and Legionella, and the abundance of enteric pathogens was significantly less than the environmental pathogens, mainly, Citrobacter, Enterococcus, Escherichia-Shigella, Enterobacter, Bacteroides. The qPCR results showed that the 16S rRNA genes of Vibrio were the most abundant, with concentrations between 7.06 and 9.48 lg copies/L, followed by oaa gene, fliC gene, trh gene, and uidA gene, and the temperature and salinity were the main factors affecting its abundance. Variance partitioning analysis (VPA) analysis of spatial factors on the potential pathogen’s distribution (19.6% vs 5.3%) was greater than environmental factors. In addition, the co-occurrence analysis of potential pathogens in the estuary revealed significant co-occurrence among the opportunistic pathogens Testosteronemonas, Brevimonas vesicularis, and Pseudomonas putida. Our findings provide an essential reference for monitoring and occurrence of potentially pathogenic bacteria in estuaries.