Public health aspects of Vibrio spp. related to the consumption of seafood in the EU

EFSA Journal, Journal Year: 2024, Volume and Issue: 22(7)

Published: July 1, 2024

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Deployment of a Vibrio cholerae ordered transposon mutant library in a quorum-competent genetic background

mBio, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

ABSTRACT Vibrio cholerae , the causative agent of cholera, has sparked seven pandemics in recent centuries, with current one being most prolonged. V. cholerae’s pathogenesis hinges on its ability to switch between low- and high-cell-density gene regulatory states, enabling transmission host environment. Previously, a transposon mutant library for was created support investigations aimed toward uncovering genetic determinants pathogenesis. However, subsequent sequencing uncovered mutation luxO parent strain, rendering mutants unable exhibit behaviors. In this study, we used chitin-independent natural transformation move insertions from these low-cell-density into wild-type genomic background. Library transfer aided by novel gDNA extraction method developed using thymol, which also showed high lysis specificity . The resulting Grant comprises 3,102 unique mutants, covering 79.8% open reading frames. Whole-genome randomly selected demonstrates 100% precision cognate positions recipient strain every analyzed. Notably, no instance did Our research density-dependent epistasis growth inosine, an immunomodulatory metabolite secreted gut bacteria that is implicated enhancing barrier functions. Additionally, retain plasmid enables rapid, scarless editing. summary, reintroduces organismal-relevant contexts absent low-cell-density-locked equivalent. Ordered libraries are essential tools catalyzing providing access null all non-essential genes. Such previously generated but whole-genome revealed made cell-cell communication known as quorum sensing. Here, utilize competence combined novel, high-throughput DNA regenerate signaling incompetent ordered quorum-sensing-competent strain. provides researchers powerful tool understand biology within context influences how it transitions environmentally benign organism disease-causing pathogen.

Language: Английский

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Punicalagin inhibits biofilm formation and virulence gene expression of Vibrio parahaemolyticus

Food Control, Journal Year: 2022, Volume and Issue: 139, P. 109045 - 109045

Published: April 26, 2022

Language: Английский

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Transcriptomic analysis reveals the antibiofilm mechanism of Lacticaseibacillus rhamnosus MS1 against Vibrio parahaemolyticus

LWT, Journal Year: 2023, Volume and Issue: 176, P. 114529 - 114529

Published: Feb. 1, 2023

This study aimed to explore novel anti-biofilm agents from Lacticaseibacillus rhamnosus. We found that the ethyl acetate extract of L. rhamnosus MS1 effectively reduced motility, extracellular polysaccharide (EPS) content and biofilm production Vibrio parahaemolyticus confirmed effect by light microscopy scanning electron microscopy. Transcriptomic analysis revealed there were 284 significantly up-regulated genes 443 down-regulated in V. treated with subinhibitory concentration MS1. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted verify expression changes biofilm-related (cpxA, cysD, D0853_RS09350, D0853_RS15790, emrD, flgM, luxS, ompW, pyk tdh), results indicated regulated quorum sensing (QS), formation, virulence factors related metabolic pathways. efficacy as an agent.

Language: Английский

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Characterization of GefA, a GGEEF Domain-Containing Protein That Modulates Vibrio parahaemolyticus Motility, Biofilm Formation, and Virulence

Applied and Environmental Microbiology, Journal Year: 2022, Volume and Issue: 88(6)

Published: Feb. 2, 2022

Vibrio parahaemolyticus is a significant foodborne pathogen that causes economic and public health problems worldwide has high capacity to adapt diverse environments hosts. The second messenger cyclic diguanylate monophosphate (c-di-GMP) allows bacteria shift from planktonic form communal multicellular lifestyle plays an important role in bacterial survival transmission. Here, we characterized single-domain c-di-GMP synthetases V. identified novel GGEEF domain-containing protein designated GefA modulates swarming motility, biofilm formation, virulence. inhibits motility by regulating the expression of lateral flagella, while it enhances formation controlling exopolysaccharide biosynthesis. Under high-c-di-GMP conditions caused scrABC knockout, found bifunctional, as no effect on but retains ability regulate formation. Subsequent studies suggested regulates type III secretion system 1 (T3SS1), which virulence factor parahaemolyticus. also revealed flagella participate infection We both T3SS1 contribute GefA-mediated zebrafish model. Our results expand knowledge their roles social behaviors pathogenicity. IMPORTANCE metabolic enzymes constitute one largest clusters potential orthologues However, specific these individual play are largely unknown. demonstrated this bacterium, through To our knowledge, have never been revealed. findings better understanding function its

Language: Английский

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Early stage of biofilm assembly on microplastics is structured by substrate size and bacterial motility

iMeta, Journal Year: 2023, Volume and Issue: 2(3)

Published: June 7, 2023

The taxonomic structure of biofilms on 0.3-mm microplastics differed significantly from that 3-mm or glass particles. Compared with the microplastics, were enriched for genes involved in flagellar-based motility and chemotaxis, pointing to a more 'mobile' community. association between bacterial colonization was observed through laboratory experiments using isolated strains. Microplastic particles (MPs), defined as plastic debris diameters less than 5 mm, are dominant form plastics oceans earth [1]. These have become one most severe threats entire marine ecosystem, largely due poor biodegradation harmful chemical compounds, many which persistent organic pollutants [2-4]. abundant type MPs is polyethylene, followed by polypropylene, polyvinyl chloride, polystyrene [5]. hydrophobic surfaces allow biofilm formation. Such MP surface 'plastisphere', first described during study North Atlantic [6]. Thus, 'plastisphere' represents specific community, can be found nearly all substrates immersed seawater, including animal body surfaces, rock particles, sediment–water interfaces [7]. Biofilm formation can, turn, promote spread [8]. First, lead increased density and, thus reduced buoyancy Second, matrix sticky heteroaggregates also impact element cycling example, presence has been shown alter nitrogen processes [9]. reports demonstrate significance MP-associated microbiota environmental effects MPs, indicating mechanisms governing assembly great importance. Various parameters known determine structures [10-16]. For pointed out microbial diversity uniformity higher those they co-influenced polymer types exposure duration [11]. A factors influencing suggested seawater salinity main factor [12]. Interestingly, color affect both functional composition [13]. Moreover, relationship size (diameter) properties investigated few studies [14-16]. wastewater treatment systems, influences behavior antibiotic-resistance [14]. However, influence situ environments remain elusive. In present study, we conducted experiment coastal area, employing (GPs) different sizes immersing these materials subtidal zone 10 20 days before recovery. Subsequently, metagenomics, 16S rRNA gene amplicon analyses, metatranscriptomics, quantitative real-time polymerase chain reaction (qRT-PCR) performed elucidate uniqueness formed MPs. Furthermore, based culturable strains support our conclusion. experimental design Figure 1A. Before deploy into characters scanning electron microscope (SEM), no apparent difference could identified (Figure S1). total, 27 samples collected metagenomic sequencing generated total 554.09 Gb data sets 20.52 ± 1.88 per sample (Table 10-day 20-day termed 3-MP-10 3-MP-20, respectively. 0.3-MP-10 0.3-MP-20, GPs 3-GP-10, 3-GP-20, 0.3-GP-10, 0.3-GP-20, following same naming rule. Operational unit (OTU) classification miTags extracted metagenomes at 97% similarity revealed 18,356 OTUs. Taxonomic assignments representative reads OTUs 78 phyla (Proteobacteria further classified class level), phylum-level community displayed 1B. profiles biofilms, Alphaproteobacteria emerging taxa, while Betaproteobacteria, Actinobacteria, Crenarchaeota made up relatively larger proportion comparison their counterparts 1B). Gammaproteobacteria Bacteroidetes prevalent taxa across biofilms. Among assayed, unique, accounting 61.57%, fewer detected other addition, there appeared structural transition 0.3-MP-20 indicated 3.0-fold decline 3.6-fold rise relative abundances Bacteroidetes, respectively At genus level, 0.3-MP clearly 1C). genera, Oleiphilus Oleibacter, accounted averages 14.71% 14.35%, respectively, showing abundance Alteromonas microbiota. seen elimination development results unique earlier-stage 0.3-MPs. dissimilarities then principal coordinate analysis (PCoA). 20-day-immersed analyzed separately Bray-Curtis Jaccard distances. an boundary S2). PCoA samples, located Similarly, Overall, plots especially consistent profiling results. As above analyses miTags, used (spanning V3V4 regions) verify Data information given Table S2. Each contained 50,000 sequences, rarefaction curves Chao1 Shannon indexes depth sufficient richness S3). Consistent miTag 1B), sequences dominance increases S4). Oleiphilus, Nautella, S5), Although not separate distinguish S6), suggesting (identity abundance). Together, derived verified led us explore specification. assembled individually, open reading frames (ORFs) predicted annotated Kyoto Encyclopedia Genes Genomes (KEGG) database. Of 195,721 47,617 ORFs, 5493 218 ORFs After read mapping summary, each KEGG revealed. Considering subsequent one-way variance (ANOVA) identify comparative four groups. We 3758 6518 KEGG-annotated changed (adjusted p-value < 0.001 ANOVA), 749 Classification categories, among categories highest numbers S7). significantly-changed genes, top 40 terms F-value ANOVA, S8. Notably, seven related flagellar biosynthesis regulation, such flgK: encoding hook-associated protein 1 (K02396) fliK: hook-length control (K02414), 2A). "mobile" early stages To confirm result compared DESeq2 analysis, number queries. 1460 0.001). Nearly flagellar-related showed significant (fold change > 2) S9). fliH largest fold Other fliC, flgE, motA, two-fold increase explored affiliations fliC flgE metagenomes, after assembly, ORF prediction, annotation. common bacteria belonging Alteromonas, Cognatishimia, certain Roseobacter group members (e.g., Sulfitobacter, Roseovarius, Ruereria) S10). motility-related metatranscriptomics examine expression metatranscriptome created each. basic information, length, provided S3. levels metatranscriptomic metagenome-derived orthologs. overall expression-level ranks 2B,C, Distinct distribution patterns observed, tending up-left area close y-axis (representing 0.3-MP-10) tended bottom-right x-axis 3-MP-10) 2B). Consistently, rank 2C). Then took closer look two metatranscriptomes individually 2D). documented biofilm, evident fliK flgF (K02391), (K02406), mcp (K03406) connection understand active mobile microbes S11). subjected prediction annotation function taxonomy. Bacteria Pseudoalteronomas, Thalassolituus, major hosts results, qRT-PCR 3-MP Its various (Alteromonas, Leisingera, Phaeobacter, Vibrio) (5, 10, 15 days) examined. selected metatranscriptome-derived (respective primers It expressed five genera but only (Alteromonas S12). stage-specific pattern hypothesize high We, therefore, single strain test this hypothesis. month identification selection suitable experiments. 350 stains (genus-level S13) obtained firstly candidate strain. Members Oleibacter considered difficulties culturing hindered complete genome sequenced. possess rRNAs, 67 tRNAs, 3743 1805 database S5). Whole-genome searching against GTDB-Tk bacterium sp009811495, it named A. sp009811495 PMMA93 (PMMA93 identity assigned culture). Analysis its metabolic pathways focus energy metabolism pathway complete, comprising about 34 proteins associated motor, stator, basal body, proximal rod, rings, hook, filament, cap S14). possessed Embden-Meyerhof-Parnas tricarboxylic acid cycle S14), heterotrophic organism. nitrate nitrite respiration potential living without oxygen. responsible well relevant regulatory cell membrane-supported respiratory provide required swimming swarming. ability PMMA93, assay broth 2216E agar plates. Another strain, Polaribacter dokdonensis PC73 negative (the metagenomes). On plates 0.3% (w/v) agar, occupied almost whole plate 25°C, whereas sign S15). 0.5% swarm days, loose colony diameter 2.7 cm assess contribution cultured cells 25°C log phase, treated them concentrations carbonyl cyanide 3-chlorophenylhydrazone (CCCP), compound eliminate proton motive force (PMF), incubated 3-MPs 0.3-MPs study. 12 h, colonized washed SEM observation 3A), optical measurement 600 nm (OD600) 3B), counting colonies plated media 3C). CCCP exerted dose-dependent 12.5 µΜ 25 abrogating over 50% 75% cells, 3A–C). contrast, investigate coculture motile (A. PMMA93) nonmotile (Polaribacter sp. PC73). Cells mixed 1:1 ratio h. This biofilm-associated (cells MPs) free-living (in media) phases. Information S6. Subsequent approximately phases, regardless S16A). percentage decreased 29%–37% 10%–15% Statistical (t-test, 0.01) S16B). S16A,B). colonize effectively. finding, examined Three Vibrio alginolyticus 2-8, Stentrophomonas pavanii P5G5, Tritonibacter mobilis 2-12, experiment. Similar three abolished CCCP, slight, changes S17). Finally, correlation colonization, knocked motAB (primer S7) within lateral cluster V. 2–8 colonizing mutant wild-type defect 0.3-MPs, S18). microplastic zone. Based earlier stage (10-day development), had structures. enrichment motivated perform successful plastisphere "just" normal special stone surfaces) [17]. view supported finding differences communities usually driven rare groups unchanged [18, 19], variables often much [20, 21]. here demonstrated stage, represented Oleibacter. oil droplets [22] contains giant flagellins thick filaments, speculated facilitate adaptation niches [23]. Therefore, highlighted structurally explained increasing intra-community forces development. observations previous [24-26]. zone, age stronger effect shaping substrate [25]. imply might involve species-species interactions reduce replacement pioneer species Oleiphilus). consistent, approaches address characteristics plastisphere. Both metagenomics metatranscriptomica days. Mechanistically, physical features attachment encounters hydrodynamic when water. Hydrodynamic arising water velocity Brownian movement suspended small partially arrested swimming, provides "kinetic force" counteract force, preventing nonmobile [27]. explains well-known capability degrade consume require overcome hydrostatic oil-seawater interfaces. mechanism recent enhanced adhesion Halomonas surfactant-coated dispersed artificial [28]. unlike electrostatically charged [29], may difficulty cause GPs, several 0.3-GP-10 3-GP-10 areas contribute greater chance contacting confined space, bag containing experimentation Besides mechanisms, prevalence chemotaxis-related implies possibility compounds released play roles recruiting particular species. additives oligopolymers [30] likely consumed taxa. notion line conclusion [31], authors chemically particle consumption contributed assembly. underwent rapid turnover, developing capable degrading carbon 140 h case current MP-released chemicals need studied. corroborate field experiments, microbiological importance colonization. Because PMFs [32], suggest probably energy-consuming process, complex cellular process requires investment host [33]. competitiveness understanding leads competitive advantage nutrient patches [34]. cultivation faster propagation rate mutation cluster, specifically swarming rather liquid medium [35], confirmed role findings There limitations location fluctuations considered. [10, 12], underlying basis understood, would future directions. analyses. complement limitation, conducted, grown shortcoming development, currently attempting construction synthetic hundreds strains, model decipher strain- single-gene level Combining meta-analyses propose structured motility, creates communities. refreshed microbe-surface possibly underpin ecological transport environments. Weipeng Zhang Wei Ding conceived project. Peng Qin Han Cui part Jinshui Lin, Panxin Li, Shuaitao Wang Shen Fan, Jie Lu, Meng Sun, Heng Zhang, Shougang Wang, Xiaoyan Su Yu-Zhong Xiaoli Hu, Hui-Hui Fu technical comments. wrote manuscript. work overseas top-level innovation talents funding (862105020028 862205033005) Zhang. grateful Jinshan Tan Qingdao University observation. declare conflict interest. New article. metagenomic, amplicon, uploaded NCBI under accession PRJNA902427 (https://www.ncbi.nlm.nih.gov/sra/PRJNA902427). SAMN32034847 (https://www.ncbi.nlm.nih.gov/nuccore/CP113972). Supplementary (methods, figures, tables, graphical abstract, Chinese translated version update materials) online DOI iMeta Science http://www.imeta.science/. Please note: publisher content functionality any supporting supplied authors. Any queries (other missing content) should directed corresponding author

Language: Английский

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Antibacterial and antibiofilm activities of iodinated hydrocarbons against Vibrio parahaemolyticus and Staphylococcus aureus

Scientific Reports, Journal Year: 2024, Volume and Issue: 14(1)

Published: April 22, 2024

Abstract Food-related illnesses have become a growing public concern due to their considerable socioeconomic and medical impacts. Vibrio parahaemolyticus Staphylococcus aureus been implicated as causative organisms of food-related infections poisoning, both can form biofilms which confer antibiotic resistance. Hence, the need for continuous search compounds with antibiofilm antivirulence properties. In this study, 22 iodinated hydrocarbons were screened activity, these, iodopropynyl butylcarbamate (IPBC) was found effectively control biofilm formation pathogens MIC 50 µg/mL bactericidal V. S. . Microscopic studies confirmed IPBC inhibits bacteria also disrupted mixed formation. Furthermore, suppressed virulence activities such motility hemolytic activity cell surface hydrophobicity It exhibited preservative potential against in shrimp model. membrane differentially affected gene expressions related virulence. Additionally, it displayed broad-spectrum other clinically relevant pathogens. These findings indicate offers means controlling mediated by biofilms.

Language: Английский

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Author response: Polysaccharide breakdown products drive degradation-dispersal cycles of foraging bacteria through changes in metabolism and motility

Published: July 24, 2024

Most of Earth's biomass is composed polysaccharides. During decomposition, polysaccharides are degraded by heterotrophic bacteria as a nutrient and energy source thereby partly remineralized into CO2. As heterogeneously distributed in nature, following the colonization degradation polysaccharide hotspot cells need to reach new hotspots. Even though many studies indicate that these degradation-dispersal cycles contribute carbon flow marine systems, we know little about how alternate between motility, which environmental factors trigger this behavioral switch. Here, studied growth bacterium Vibrio cyclitrophicus ZF270 on abundant alginate, both its soluble polymeric form well breakdown products. We used microfluidics coupled time-lapse microscopy analyze motility individual cells, RNA sequencing study associated changes gene expression. found single grow at reduced rate alginate until they large groups cooperatively break down polymer. Exposing cell digested accelerates expression genes involved catabolism, central metabolism, ribosomal biosynthesis, transport. However, exposure also triggers become motile disperse from groups, proportionally increasing with group size before switch, accompanied high flagellar assembly, chemotaxis, quorum sensing. chemotax toward but not likely enabling them find Overall, our findings reveal cellular mechanisms might underlie bacterial cycles, influence remineralization environments.Polysaccharides, known glycans, most Earth understanding microorganisms essential for global cycle storage release CO2 natural systems. Although formation common strategy degrade ubiquitous substrates where hotspots distributed, prepare dispersal an exhausted re-initiate patches. By quantifying growth, chemotaxis comparing when populations were exposed either or their products polysaccharides, show alter behavior experience shift polysaccharides: After during polymers, triggered motile, sessile groups. The chemotactic attraction allows move towards Our sheds light processes drive remineralization, important process resulting

Language: Английский

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H-NS represses fliD transcription for lateral flagellar filament cap in Vibrio parahaemolyticus

Archives of Microbiology, Journal Year: 2025, Volume and Issue: 207(2)

Published: Jan. 9, 2025

Language: Английский

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Antimicrobial Efficacy of Trifluoro-Anilines Against Vibrio Species

International Journal of Molecular Sciences, Journal Year: 2025, Volume and Issue: 26(2), P. 623 - 623

Published: Jan. 13, 2025

Vibrios are naturally present in marine ecosystems and commonly allied with live seafood. Vibrio species frequently cause foodborne infections, parahaemolyticus recently becoming a significant contributor to illness outbreaks. In response, aniline 68 of its derivatives were studied due their antibacterial effects targeting V. harveyi. Among these, 4-amino-3-chloro-5-nitrobenzotrifluoride (ACNBF) 2-iodo-4-trifluoromethylaniline (ITFMA) demonstrated both antibiofilm properties. The minimum inhibitory concentrations (MIC) for ACNBF ITFMA 100 µg/mL 50 µg/mL, respectively, against planktonic cells. active compounds effectively suppressed biofilm formation manner dependent on the dosage. Additionally, these trifluoro-anilines significantly reduced virulence factors such as motility, protease activity, hemolysis, indole production. Both caused noticeable destruction membrane bacterial cells and, at exhibited bactericidal activity within 30 min. Toxicity assays using Caenorhabditis elegans seed germination models showed that displayed mild toxicity. As result, inhibited growth formation. Furthermore, prevented surfaces shrimp squid models, highlighting potential use controlling seafood contamination.

Language: Английский

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