Abstract

BackgroundVibrio parahaemolyticus is a main causative agent of serious human seafood-borne gastroenteritis disease. Many researchers have investigated its pathogenesis by observing the alteration of its virulence factors in different conditions. It was previously known that culture conditions will influence the gene expression and the metabolic profile of V. parahaemolyticus, but little attention has been paid on the relationship between them. In this study, for the first time, the metabolomics response in relation to the expression of two major virulence genes, tdh and trh, induced at three temperatures (4, 25 and 37 °C) was examined in two genotypes of pathogenic Vibrio parahaemolyticus (ATCC33846 (tdh+/trh−/tlh+) and ATCC17802 (tdh−/trh+/tlh+)).ResultsReverse transcription real-time PCR (RT-qPCR) analysis illustrated that the expression levels of tdh and trh induced at 25 °C in V. parahaemolyticus were significantly higher than those induced at 4 and 37 °C. Principal components analysis (PCA) based on the UPLC & Q-TOF MS data presented clearly distinct groups among the samples treated by different temperatures. Metabolic profiling demonstrated that 179 of 1,033 kinds of identified metabolites in ATCC33846 changed significantly (p <0.01) upon culturing at different temperatures, meanwhile 101 of 930 kinds of metabolites changed (p <0.01) in ATCC17802. Pearson’s correlation analysis highlighted the correlation between metabolites and virulence gene expression levels. At the threshold of | r | = 1, p <0.01, 12 kinds of metabolites showed extremely significant correlations with tdh expression, and 4 kinds of metabolites significantly correlated with trh expression. It is interesting that 3D, 7D, 11D-Phytanic acid showed the same trend with pyrophosphate, whose derivative could activate the degradation of phytanic acid. Several metabolites could be sorted into the same class by the method of chemical taxonomy, by assuming that they are involved in the same metabolic pathways.ConclusionsThis research can help to find biomarkers to monitor virulence gene expression, and can further help laboratory and clinical research of V. parahaemolyticus from the perspective of metabolomics.Electronic supplementary materialThe online version of this article (doi:10.1186/s12866-016-0688-5) contains supplementary material, which is available to authorized users.

Highlights

  • Vibrio parahaemolyticus is a main causative agent of serious human seafood-borne gastroenteritis disease

  • Virulence gene expression of V. parahaemolyticus The virulence gene expression of two genotypes of V. parahaemolyticus were investigated by retention time (RT)-qPCR under different culture conditions (4, 25 and 37 °C)

  • Both pvuA and pvsA were used as the reference genes due to the bias which may be caused by the fluctuation in expression level of a single reference gene [23]

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Summary

Introduction

Vibrio parahaemolyticus is a main causative agent of serious human seafood-borne gastroenteritis disease. For the first time, the metabolomics response in relation to the expression of two major virulence genes, tdh and trh, induced at three temperatures (4, 25 and 37 °C) was examined in two genotypes of pathogenic Vibrio parahaemolyticus (ATCC33846 (tdh+/trh−/tlh+) and ATCC17802 (tdh−/trh+/tlh+)). V. parahaemolyticus strains contain a number of different virulence factors including adhesins, thermostable direct hemolysin (TDH), TDH-related hemoysin (TRH), two type III secretion systems, T3SS1 and T3SS2 [8, 9]. Previous studies have found that pathogenic V. parahaemolyticus often carries thermostable direct hemolysin (tdh) and/or thermostable-related hemolysin (trh) genes [12,13,14]. Studies of virulence factors have made remarkable progress, while the synergy effect and pathogenicity of them are still under investigation [9]

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