Proteomics and 1H NMR-based metabolomics analysis of pathogenic Vibrio vulnificus aquacultures isolated from sewage drains
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Vibrio bacteria live in both marine and freshwater habitats and are associated with aquatic animals. Vibrio vulnificus is a pathogenic bacterium that infects people and livestock. It is usually found in offshore waters or within fish and shellfish. This study presents a comparative proteomic analysis of the outer membrane protein (OMP) changes in V. vulnificus proteins after stimulation with sewage from sewage drains. Using two-dimensional electrophoresis followed by MALDI-TOF MS/MS, 32 protein spots with significant differences in abundance were identified and characterized. These identified proteins were found to be involved in various functional categories, including catalysis, transport, membrane proteins progresses, receptor activity, energy metabolism, cytokine activity, and protein metabolism. The mRNA expression levels of 12 differential proteins were further assessed by qRT-PCR. Seven genes including carboxypeptidase, hemoglobin receptor, succinate dehydrogenase iron-sulfur subunit, ATP synthase subunit alpha, thioredoxin, succinyl-CoA synthetase subunit, and alanine dehydrogenase were downregulated upon stimulation, whereas the protein expression levels HupA receptor, type I secretion outer membrane protein, glutamine synthetase, superoxide dismutase, OmpU, and VuuA were upregulated. 1H NMR spectra showed 18 dysregulated metabolites from V. vulnificus after the sewage stimulation and the pathogenicity was enhanced after that.
KeywordsVibrio vulnificus Sewage Proteomics Metabolomics 1H NMR qRT-PCR
This work was supported by the Public Science and Technology Research Funds ocean projects (201105007, 201005016), the Regional Demonstration Project of Marine Economic Innovation and Development (2014, 2016), the Scientific Research Foundation of the Graduate School of Ningbo University, and K.C. Wong Magna Fund in Ningbo University. We thank the American Journal Experts (AJE) for English language editing. This manuscript was edited for English language by the American Journal Experts (AJE).
Compliance with ethical standards
This article does not contain any studies with human participants or animals performed by any of the authors.
Conflict of interest
The authors declare that they have no conflicts of interest.
- 3Rd GJ, Park Y, Lamers Y, Bandyopadhyay N, Chi YY, Lee K, Kim S, Da SV, Hove N, Ranka S (2013): Metabolomic analysis reveals extended metabolic consequences of marginal vitamin B-6 deficiency in healthy human subjects. PLoS One 8, e63544Google Scholar
- Bisharat N, Agmon V, Finkelstein R, Raz R, Ben-Dror G, Lerner L, Soboh S, Colodner R, Cameron DN, Wykstra DL (1999) Clinical, epidemiological, and microbiological features of Vibrio vulnificus biogroup 3 causing outbreaks of wound infection and bacteraemia in Israel. Lancet 354:1421–1424CrossRefGoogle Scholar
- Bollard ME, Holmes E, Lindon JC, Mitchell SC, Branstetter D, Zhang W, Nicholson JK (2001) Investigations into biochemical changes due to diurnal variation and estrus cycle in female rats using high-resolution (1)H NMR spectroscopy of urine and pattern recognition. Anal Biochem 295:194–202CrossRefGoogle Scholar
- Davey ML, Hancock RE, Mutharia LM (1998) Influence of culture conditions on expression of the 40-kilodalton porin protein of Vibrio anguillarum serotype O2. Applied & Environmental Microbiology 64:138–146Google Scholar
- Davis SR, Quinlivan EP, Stacpoole PW (2006) Plasma glutathione and cystathionine concentrations are elevated but cysteine flux is unchanged by dietary vitamin B-6 restriction in young men and women. J Nutr 136:373–378Google Scholar
- Davis W, Ronai ZE, Tew KD (2001) Cellular thiols and reactive oxygen species in drug-induced apoptosis. Journal of Pharmacology & Experimental Therapeutics 296:1–6Google Scholar
- Depaola A, Capers GM, Alexander D (1994) Densities of Vibrio vulnificus in the intestines of fish from the U.S. Gulf Coast. Applied & Environmental Microbiology 60:984–988Google Scholar
- Horseman MA, Surani S (2010) A comprehensive review of Vibrio vulnificus: an important cause of severe sepsis and skin and soft-tissue infection. International Journal of Infectious Diseases Ijid Official Publication of the International Society for Infectious Diseases 15:e157–e166CrossRefGoogle Scholar
- Jiang N, Chun-Li LI, Zhou J, Wang ZH, Tai-Wu LI, Xiu-Rong SU (2013) Diversity and distribution of dominant bacteria nearby Ningbo land-based drainage discharge outlets. Journal of Ningbo UniversityGoogle Scholar
- Kim YR, Kim BU, Kim SYKim CM, Na HS, Koh JT, Choy HE, Rhee JH, Lee SE (2010): Outer membrane vesicles of Vibrio vulnificus deliver cytolysin-hemolysin VvhA into epithelial cells to induce cytotoxicity. Biochemical & Biophysical Research Communications 399, 607–612Google Scholar
- Long S, Mothibeli MA, Robb FT, Woods DR (1981) Regulation of extracellular alkaline protease activity by histidine in a collagenolytic Vibrio alginolyticus strain. J Gen Microbiol 127:193–199Google Scholar
- Okujo N, Akiyama T, Miyoshi S, Shinoda S, Yamamoto S (1996): Involvement of vulnibactin and exocellular protease in utilization of transferrin- and lactoferrin-bound iron by Vibrio vulnificus 40, 595–598Google Scholar
- Pei Z, Burucoa C, Grignon B, Baqar S, Huang XZ, Kopecko DJ, Bourgeois AL, Fauchere JL, Blaser MJ (1998) Mutation in the peb1A locus of Campylobacter jejuni reduces interactions with epithelial cells and intestinal colonization of mice. Infection & Immunity 66:938Google Scholar
- Porcheron G, Garénaux A, Proulx J, Sabri M, Dozois CM (2012) Iron, copper, zinc, and manganese transport and regulation in pathogenic Enterobacteria: correlations between strains, site of infection and the relative importance of the different metal transport systems for virulence. Frontiers in Cellular & Infection Microbiology 3:90Google Scholar
- Søorum H, Poppe TT, Olsvik O (1988) Plasmids in Vibrio salmonicida isolated from salmonids with hemorrhagic syndrome (Hitra disease). J Clin Microbiol 26:1679Google Scholar
- Surasilp T, Longyant S, Rukpratanporn S, Sridulyakul P, Sithigorngul P, Chaivisuthangkura P (2011) Rapid and sensitive detection of Vibrio vulnificus by loop-mediated isothermal amplification combined with lateral flow dipstick targeted to rpoS gene. Mol Cell Probes 46:1122–1131Google Scholar
- Volker Behrends TJB, Manuel Liebeke, Anne Cordes-Blauert, Syedah N. Ashraf, Chandrika Nair, James E. A. Zlosnik, Huw D. Williams, Jacob G. Bundy (2013): Metabolite profiling to characterize disease-related bacteria, gluconate excretion by Pseudomonas aeruginosa mutants and clinical isolates from cystic fibrosis patients. J Biol Chem 288, 15098–15109Google Scholar
- Wright AC, Simpson LM, Oliver JD (1981) Role of iron in the pathogenesis of Vibrio vulnificus infections. Infection & Immunity 34:503–507Google Scholar
- Yeom J, Shin JH, Yang JY, Kim J, Hwang GS (2013): (1)H NMR-based metabolite profiling of planktonic and biofilm cells in Acinetobacter baumannii 1656-2. Plos One 8, e57730Google Scholar
- Zhang D, Shan H, Ming T, Lu C, Zhou J, Su X (2017): A metabonomic analysis on the response of Enterobacter cloacae from coastal outfall for land-based pollutant under phoxim stress. Archives of microbiology, 1-9Google Scholar