Diverse Aquatic Animal Matrices Play a Key Role in Survival and Potential Virulence of Non-O1/O139 Vibrio cholerae Isolates

Abstract

Vibrio cholerae can cause pandemic cholera in humans. The waterborne bacterium is frequently isolated from aquatic products worldwide. However, current literature on the impact of aquatic product matrices on the survival and pathogenicity of cholerae is rare. In this study, the growth of eleven non-O1/0O139 V. cholerae isolates recovered from eight species of commonly consumed fish and shellfish was for the first time determined in the eight aquatic animal matrices, most of which highly increased the bacterial biomass when compared with routine trypsin soybean broth (TSB) medium. Secretomes of the V. cholerae isolates (draft genome size: 3,852,021–4,144,013 bp) were determined using two-dimensional gel electrophoresis (2DE-GE) and liquid chromatography-tandem mass spectrometry (LC-MS/MS) techniques. Comparative secretomic analyses revealed 74 differential extracellular proteins, including several virulence- and resistance-associated proteins secreted by the V. cholerae isolates when grown in the eight matrices. Meanwhile, a total of 8,119 intracellular proteins were identified, including 83 virulence- and 8 resistance-associated proteins, of which 61 virulence-associated proteins were absent from proteomes of these isolates when grown in the TSB medium. Additionally, comparative genomic and proteomic analyses also revealed several strain-specific proteins with unknown functions in the V. cholerae isolates. Taken, the results in this study demonstrate that distinct secretomes and proteomes induced by the aquatic animal matrices facilitate V. cholerae resistance in the edible aquatic animals and enhance the pathogenicity of the leading waterborne pathogen worldwide.

Advances in cholera research: from molecular biology to public health initiatives

“…Zebrafish ( Danio rerio ) have been shown to be naturally susceptible to infection and colonization by V. cholerae and have been developed as a V. cholerae model ( Runft et al, 2014 ; Mitchell et al, 2017 ; Mitchell and Withey, 2018 ; Nag et al, 2018b , 2020 ). Aquatic reservoirs harboring both environmental and pandemic V. cholerae strains provide rich conditions for genetic evolution, and many non-O1/non-O139 have been found to contain partial pathogenicity islands and fully intact virulence genes, including the Vibrio seventh pandemic islands 1 and 2 (VSP-1, VSP-2), toxin co-regulated pilin ( tcpA ), hemolysin A ( hlyA ), and the Type 6 secretion system (T6SS) ( Li et al, 2014 , 2019a ; Yan et al, 2022 ; Santoriello et al, 2023 ).…”

Comparative secretomic and proteomic analysis reveal multiple defensive strategies developed by Vibrio cholerae against the heavy metal (Cd2+, Ni2+, Pb2+, and Zn2+) stresses

“…The V. cholerae isolates were treated with the heavy metals for 2 h as described in the Scanning Electron Microscopy (SEM) Analysis section, but incubated without shaking. Extracellular proteins of the V. cholerae isolates were extracted as described in our recent reports (Zhu et al, 2020;Shan et al, 2022;Yan et al, 2022). Isoelectric focusing (IEF) was performed using immobilized pH gradient (IPG) gels (pH 4-7, 7 cm; Bio-Rad, Hercules, USA).…”

“…In our previous studies, the 2D-GE combined with LC-MS/MS techniques were also applied in global identification of DEPs in Vibrio species (Zhu et al, 2020;Shan et al, 2022;Yan et al, 2022). For example, Zhu et al (2020) compared secretomes and proteomes of Vibrio parahaemolyticus strains isolated from 12 species of aquatic animals and identified 28 differential extracellular proteins.…”

Bacterial nucleoid-associated protein HU as an extracellular player in host-pathogen interaction

“…Escherichia coli eDNA; LPS; biofilm (Devaraj et al, 2015;Thakur et al, 2021) Francisella tularensis culture supernatant; vesicles (Konecna et al, 2010;Klimentova et al, 2019) Mycobacterium tuberculosis extracellular; binds host polysaccharide (Aoki et al, 2004) Aggregatibacter actinomycetemcomitans biofilm; interacts with host cytokine IL-1b (Paino et al, 2012;Freire et al, 2017) Wolbachia host cell (Beckmann et al, 2013) Helicobacter pylori culture supernatant (Kim et al, 2002) Vibrio cholera culture supernatant (Yan et al, 2022) Streptococcus intermedius culture supernatant (Liu et al, 2008) group A streptococci interacts with heparin and lipoteichoic acid (Choi and Stinson, 1991;Winters et al, 1993;Stinson et al, 1998) Streptococcus pyogenes surface-exposed; culture supernatant (Lei et al, 2000;Severin et al, 2007) Haemophilus influenzae biofilm (Brockson et al, 2014;Jurcisek et al, 2017) Pseudomonas aeruginosa biofilm (Gustave et al, 2013;Novotny et al, 2016) Burkholderia cenocepacia biofilm (Novotny et al, 2013) Porphyromonas gingivalis biofilm (Rocco et al, 2017) Streptococcus gordonii biofilm (Rocco et al, 2017) together with DNA in the early stages of attachment to the surface and becomes part of bacterial biofilm (Jurcisek et al, 2017). HU protein can be released also by other mechanism, including explosive lysis of the cells (Turnbull et al, 2016) or unknown secretion mechanisms.…”

“…HU protein was detected in culture filtrate ( Konecna et al., 2010 ) and extracellular vesicles ( Klimentova et al., 2019 ) produced by F. tularensis, with an undescribed function yet. HU protein could also contribute to the virulence of Vibrio cholerae because HU protein is secreted into the medium and thus it could contribute to the cholera disease ( Yan et al., 2022 ). Moreover, HU protein of V. cholerae is essential for CTXϕ phage replication that is responsible for virulence of V. cholerae ( Martínez et al., 2015 ; Martínez et al., 2016 ).…”