Thymoquinone Inhibits Biofilm Formation and Attachment-Invasion in Host Cells of Vibrio parahaemolyticus

Abstract Purpose: Determination of the association between ica genes and phenotypic biofilm formation in staphylococcal isolates involved in conjunctivitis, their antibiotic resistance as well as detection of selected virulence characteristics: adhesion to epithelial cells and in vitro cytotoxicity.Methods: The study included 26 Staphylococcus aureus (SA) and 26 Staphylococcus epidermidis (SE) isolates. The presence of icaAD genes and ica operon was determined by the PCR assay. Phenotypic biofilm formation was verified using the microtiter plate assay. Antibiotic resistance was performed using the disc diffusion method. Staphylococcal ability to attach to host cells was assessed by flow cytometry. Cytotoxicity on epithelial cells was evaluated by LDH assay.Results: The ica genes were detected in 26.9% of SE and in 42.3% of SA isolates. Only 15.3% of isolates (SE) were positive for both the icaAD and the ica operon. Phenotypically, 19.2% of SE isolates were strong biofilm producers, among which three were both icaAD- and ica operon-positive. 26.9% of SA isolates were strong biofilm producers. Methicillin resistance (MR) was detected in 34.6% of SE and 26.9% of SA isolates. 75% of MR isolates were multidrug resistant. SA isolates adhered to host cells more extensively than SE. SA isolates released higher level of LDH than SE.Conclusions: Adherence abilities were commonly observed in staphylococci associated with conjunctivitis. However, low prevalence of isolates positive for a complete and functional ica locus and low prevalence of strong biofilm producers was detected. SA adhered to a greater extent to eukaryotic cells than SE and were more cytotoxic.

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Biofilm formation by Vibrio parahaemolyticus on different surfaces and its resistance to sodium hypochlorite

Ciência Rural ◽

10.1590/0103-8478cr20180612 ◽

2018 ◽

Vol 48 (12) ◽

Cited By ~ 3

Author(s):

Janaina Viana da Rosa ◽

Natália Volpato da Conceição ◽

Rita de Cássia dos Santos da Conceição ◽

Cláudio Dias Timm

Keyword(s):

Stainless Steel ◽

Sodium Hypochlorite ◽

Biofilm Formation ◽

Vibrio Parahaemolyticus ◽

High Density Polyethylene ◽

High Density ◽

Bacterial Populations ◽

Farfantepenaeus Paulensis ◽

Materials Used ◽

Fish Industry

ABSTRACT: Vibrio parahaemolyticus is an important pathogen for both fish industry and consumers. It forms biofilm which makes it difficult to eliminate this microorganism using sanitizers. This study aimed to assess biofilm formation on different surfaces and effect of biofilm on resistance to sanitizers. Eight isolates of biofilm-forming V. parahaemolyticus were tested for the ability to form biofilms on a number of surfaces including high density polyethylene, stainless steel, glass, exoskeleton of Farfantepenaeus paulensis (Pink Shrimp), and operculum of Micropogonias furnieri (Whitemouth Croaker). Efficiency of sanitizer sodium hypochlorite against the bacteria was evaluated in the biofilms formed on the surface of the materials used; out the eight strains analyzed four formed biofilm on different surfaces. The present study shows that there are variations between surfaces in terms of biofilm formation, with more than one bacterial strain being able to form biofilm on the surface of the operculum of M. furnieri and on high density polyethylene as well. One isolate formed biofilm on glass, and one isolate formed biofilm on stainless steel. Sanitizers reduced biofilm formation on all surfaces. Based on our findings, we concluded that V. parahaemolyticus isolates have different ability to form biofilm on different surfaces. No isolates formed biofilm on shrimp shells. Results of this study also showed that sodium hypochlorite eat a concentration of 20 parts per million (20ppm) of Cl2, albeit not able to eliminate bacteria reported in biofilms, is still capable of reducing bacterial populations.

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Green Tea Polyphenols and Padma Hepaten Inhibit Candida albicans Biofilm Formation

Evidence-based Complementary and Alternative Medicine ◽

10.1155/2018/1690747 ◽

2018 ◽

Vol 2018 ◽

pp. 1-8 ◽

Cited By ~ 3

Author(s):

Yosi Farkash ◽

Mark Feldman ◽

Isaac Ginsburg ◽

Doron Steinberg ◽

Miriam Shalish

Keyword(s):

Candida Albicans ◽

Green Tea ◽

Crystal Violet ◽

Biofilm Formation ◽

Host Cells ◽

Green Tea Polyphenols ◽

Crystal Violet Staining ◽

Biofilm Inhibition ◽

Therapeutic Concept ◽

Biofilm Development

Candida albicans (C. albicans) is the most prevalent opportunistic human pathogenic fungus and can cause mucosal membrane infections and invade the blood. In the oral cavity, it can ferment dietary sugars, produce organic acids and therefore has a role in caries development. In this study, we examined whether the polyphenol rich extractions Polyphenon from green tea (PPFGT) and Padma Hepaten (PH) can inhibit the caries-inducing properties of C. albicans. Biofilms of C. albicans were grown in the presence of PPFGT and PH. Formation of biofilms was tested spectrophotometrically after crystal violet staining. Exopolysaccharides (EPS) secretion was quantified using confocal scanning laser microscopy (CSLM). Treated C. albicans morphology was demonstrated using scanning electron microscopy (SEM). Expression of virulence-related genes was tested using qRT-PCR. Development of biofilm was also tested on an orthodontic surface (Essix) to assess biofilm inhibition ability on such appliances. Both PPFGT and PH dose-dependently inhibited biofilm formation, with no inhibition on planktonic growth. The strongest inhibition was obtained using the combination of the substances. Crystal violet staining showed a significant reduction of 45% in biofilm formation using a concentration of 2.5mg/ml PPFGT and 0.16mg/ml PH. A concentration of 1.25 mg/ml PPFGT and 0.16 mg/ml PH inhibited candidal growth by 88% and EPS secretion by 74% according to CSLM. A reduction in biofilm formation and in the transition from yeast to hyphal morphotype was observed using SEM. A strong reduction was found in the expression of hwp1, eap1, and als3 virulence associated genes. These results demonstrate the inhibitory effect of natural PPFGT polyphenolic extraction on C. albicans biofilm formation and EPS secretion, alone and together with PH. In an era of increased drug resistance, the use of phytomedicine to constrain biofilm development, without killing host cells, may pave the way to a novel therapeutic concept, especially in children as orthodontic patients.

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Screening of a Library of Oligosaccharides Targeting Lectin LecB of Pseudomonas Aeruginosa and Synthesis of High Affinity Oligoglycoclusters

Molecules ◽

10.3390/molecules23123073 ◽

2018 ◽

Vol 23 (12) ◽

pp. 3073 ◽

Cited By ~ 3

Author(s):

Lucie Dupin ◽

Mathieu Noël ◽

Silvère Bonnet ◽

Albert Meyer ◽

Thomas Géhin ◽

...

Keyword(s):

Pseudomonas Aeruginosa ◽

Binding Sites ◽

Biofilm Formation ◽

Selective Advantage ◽

Host Cells ◽

Host Defenses ◽

Negative Bacterium ◽

High Affinity ◽

Structure Relationship ◽

Relationship Of

The Gram negative bacterium Pseudomonas aeruginosa (PA) is an opportunistic bacterium that causes severe and chronic infection of immune-depressed patients. It has the ability to form a biofilm that gives a selective advantage to the bacteria with respect to antibiotherapy and host defenses. Herein, we have focused on the tetrameric soluble lectin which is involved in bacterium adherence to host cells, biofilm formation, and cytotoxicity. It binds to l-fucose, d-mannose and glycan exposing terminal fucose or mannose. Using a competitive assay on microarray, 156 oligosaccharides and polysaccharides issued from fermentation or from the biomass were screened toward their affinity to LecB. Next, the five best ligands (Lewisa, Lewisb, Lewisx, siayl-Lewisx and 3-fucosyllactose) were derivatized with a propargyl aglycon allowing the synthesis of 25 trivalent, 25 tetravalent and 5 monovalent constructions thanks to copper catalyzed azide alkyne cycloaddition. The 55 clusters were immobilized by DNA Directed immobilization leading to the fabrication of a glycocluster microarray. Their binding to LecB was studied. Multivalency improved the binding to LecB. The binding structure relationship of the clusters is mainly influenced by the carbohydrate residues. Molecular simulations indicated that the simultaneous contact of both binding sites of monomer A and D seems to be energetically possible.

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Biofilm Formation and Methylene Blue-mediated Photodynamic Inactivation of Vibrio Parahaemolyticus in the Sea Food Industry

Nutrition & Food Science International journal ◽

10.19080/nfsij.2020.10.555787 ◽

2020 ◽

Vol 10 (3) ◽

Author(s):

Xiaoting Zhang ◽

Qian Wu ◽

Shuze Tang ◽

William W Riley ◽

Zhenqiang Chen

Keyword(s):

Methylene Blue ◽

Crystal Violet ◽

Biofilm Formation ◽

Vibrio Parahaemolyticus ◽

Food Industry ◽

Membrane Integrity ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Photodynamic Inactivation ◽

Log Reduction

This study was conducted to better understand the mechanism of Vibrio Parahaemolyticus biofilm formation and to assess the inactivation effects of methylene blue-mediated photodynamic inactivation (PDI) technology as a preventative measure. Optical microscopy, following crystal violet staining, was used to observe the kinetics of V. parahaemolyticus biofilm formation. The crystal violet-based assay was performed in microtiter plates, and it was employed to determine which factors were most influential in the formation of the biofilms. Colony counting and confocal laser scanning microscopy (CLSM) were used to test the inactivation effect of methylene blue-mediated photodynamic technology on the biofilms. V. parahaemolyticus has the ability to form biofilms, as evidenced by their immediate adherence to glass surfaces and rapid maturity, within 24 h. High (7%) or low (0.5%) salinity was not conducive to the formation of biofilms, and rotational speed greater than 130 rpm also inhibited the process. A 4.05 log reduction in the concentration of viable biofilm cells was obtained with 100 μg/mL methylene blue and 20 min irradiation (24.996 J/cm2), but planktonic cells were more susceptible to the methylene blue-mediated photodynamic reaction (5.46 log reduction). The results presented here show that the methylene blue-mediated PDI technology is an effective means to inactivate V. parahaemolyticus by disrupting its membrane integrity and to inhibit the pathogen’s formation of protective biofilms. This technology is a valid tool that can be used to enhance food safety in the sea food industry.

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Toxin-Antitoxin Genes of the Gram-Positive Pathogen Streptococcus pneumoniae: So Few and Yet So Many

Microbiology and Molecular Biology Reviews ◽

10.1128/mmbr.00030-12 ◽

2012 ◽

Vol 76 (4) ◽

pp. 773-791 ◽

Cited By ~ 36

Author(s):

Wai Ting Chan ◽

Inma Moreno-Córdoba ◽

Chew Chieng Yeo ◽

Manuel Espinosa

Keyword(s):

Streptococcus Pneumoniae ◽

Biofilm Formation ◽

Bacterial Infections ◽

Host Cells ◽

Molecular Characteristics ◽

Type Ii ◽

Pneumococcal Infections ◽

Interesting Phenomenon ◽

Content Type ◽

Toxin Protein

SUMMARYPneumococcal infections cause up to 2 million deaths annually and raise a large economic burden and thus constitute an important threat to mankind. Because of the increase in the antibiotic resistance ofStreptococcus pneumoniaeclinical isolates, there is an urgent need to find new antimicrobial approaches to triumph over pneumococcal infections. Toxin-antitoxin (TA) systems (TAS), which are present in most living bacteria but not in eukaryotes, have been proposed as an effective strategy to combat bacterial infections. Type II TAS comprise a stable toxin and a labile antitoxin that form an innocuous TA complex under normal conditions. Under stress conditions, TA synthesis will be triggered, resulting in the degradation of the labile antitoxin and the release of the toxin protein, which would poison the host cells. The three functional chromosomal TAS fromS. pneumoniaethat have been studied as well as their molecular characteristics are discussed in detail in this review. Furthermore, a meticulous bioinformatics search has been performed for 48 pneumococcal genomes that are found in public databases, and more putative TAS, homologous to well-characterized ones, have been revealed. Strikingly, several unusual putative TAS, in terms of components and genetic organizations previously not envisaged, have been discovered and are further discussed. Previously, we reported a novel finding in which a unique pneumococcal DNA signature, the BOX element, affected the regulation of the pneumococcalyefM-yoeBTAS. This BOX element has also been found in some of the other pneumococcal TAS. In this review, we also discuss possible relationships between some of the pneumococcal TAS with pathogenicity, competence, biofilm formation, persistence, and an interesting phenomenon called bistability.

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Homologous c-di-GMP-Binding Scr Transcription Factors Orchestrate Biofilm Development in Vibrio parahaemolyticus

Journal of Bacteriology ◽

10.1128/jb.00723-19 ◽

2020 ◽

Vol 202 (6) ◽

Author(s):

John H. Kimbrough ◽

J. Thomas Cribbs ◽

Linda L. McCarter

Keyword(s):

Transcription Factors ◽

Biofilm Formation ◽

Vibrio Parahaemolyticus ◽

Matrix Protein ◽

Second Messenger ◽

Minor Role ◽

Binding Motif ◽

Swarming Motility ◽

Content Type ◽

Biofilm Development

ABSTRACT The marine bacterium and human pathogen Vibrio parahaemolyticus rapidly colonizes surfaces by using swarming motility and forming robust biofilms. Entering one of the two colonization programs, swarming motility or sessility, involves differential regulation of many genes, resulting in a dramatic shift in physiology and behavior. V. parahaemolyticus has evolved complex regulation to control these two processes that have opposing outcomes. One mechanism relies on the balance of the second messenger c-di-GMP, where high c-di-GMP favors biofilm formation. V. parahaemolyticus possesses four homologous regulators, the Scr transcription factors, that belong in a Vibrio-specific family of W[F/L/M][T/S]R motif transcriptional regulators, some members of which have been demonstrated to bind c-di-GMP. In this work, we explore the role of these Scr regulators in biofilm development. We show that each protein binds c-di-GMP, that this binding requires a critical R in the binding motif, and that the biofilm-relevant activities of CpsQ, CpsS, and ScrO but not ScrP are dependent upon second messenger binding. ScrO and CpsQ are the primary drivers of biofilm formation, as biofilms are eliminated when both of these regulators are absent. ScrO is most important for capsule expression. CpsQ is most important for RTX-matrix protein expression, although it contributes to capsule expression when c-di-GMP levels are high. Both regulators contribute to O-antigen ligase expression. ScrP works oppositely in a minor role to repress the ligase gene. CpsS plays a regulatory checkpointing role by negatively modulating expression of these biofilm-pertinent genes under fluctuating c-di-GMP conditions. Our work further elucidates the multifactorial network that contributes to biofilm development in V. parahaemolyticus. IMPORTANCE Vibrio parahaemolyticus can inhabit open ocean, chitinous shells, and the human gut. Such varied habitats and the transitions between them require adaptable regulatory networks controlling energetically expensive behaviors, including swarming motility and biofilm formation, which are promoted by low and high concentrations of the signaling molecule c-di-GMP, respectively. Here, we describe four homologous c-di-GMP-binding Scr transcription factors in V. parahaemolyticus. Members of this family of regulators are present in many vibrios, yet their numbers and the natures of their activities differ across species. Our work highlights the distinctive roles that these transcription factors play in dynamically controlling biofilm formation and architecture in V. parahaemolyticus and serves as a powerful example of regulatory network evolution and diversification.

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vpaH, a Gene Encoding a Novel Histone-Like Nucleoid Structure-Like Protein That Was Possibly Horizontally Acquired, Regulates the Biogenesis of Lateral Flagella in trh-Positive Vibrio parahaemolyticus TH3996

Infection and Immunity ◽

10.1128/iai.73.9.5754-5761.2005 ◽

2005 ◽

Vol 73 (9) ◽

pp. 5754-5761 ◽

Cited By ~ 19

Author(s):

Kwon-Sam Park ◽

Michiko Arita ◽

Tetsuya Iida ◽

Takeshi Honda

Keyword(s):

Biofilm Formation ◽

Vibrio Parahaemolyticus ◽

Type Strain ◽

Wild Type Strain ◽

Enteric Bacteria ◽

Wild Type ◽

Gene Encoding ◽

Lateral Flagellum ◽

Global Gene Regulation ◽

Nucleoid Structure

ABSTRACT A histone-like nucleoid structure (H-NS) is a major component of the bacterial nucleoid and plays a crucial role in the global gene regulation of enteric bacteria. Here, we cloned and characterized the gene for the H-NS-like protein VpaH in Vibrio parahaemolyticus. vpaH encodes a protein of 134 amino acids that shows approximately 55%, 54%, and 41% identities with VicH in Vibrio cholerae, H-NS in V. parahaemolyticus, and H-NS in Escherichia coli, respectively. The vpaH gene was found in only trh-positive V. parahaemolyticus strains and not in Kanagawa-positive or in trh-negative environmental strains. Moreover, the G+C content of the vpaH gene was 38.6%, which is lower than the average G+C content of the whole genome of this bacterium (45.4%). These data suggest that vpaH was transmitted to trh-possessing V. parahaemolyticus strains by lateral transfer. The vpaH gene was located about 2.6 kb downstream of the trh gene, in the convergent direction of the trh transcription. An in-frame deletion mutant of vpaH lacked motility on semisolid motility assay plates. Western blot analysis and electron microscopy observations revealed that the mutant was deficient in lateral flagella biogenesis, whereas there was no defect in the expression of polar flagella. Additionally, the vpaH mutant showed a decreased adherence to HeLa cells and a decrease in biofilm formation compared with the wild-type strain. Introduction of the vpaH gene in the vpaH-negative strain increased the expression of lateral flagella compared with the wild-type strain. In conclusion, our findings suggest that VpaH affects lateral flagellum biogenesis in trh-positive V. parahaemolyticus strain TH3996.

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Adhesion of Campylobacter jejuni Is Increased in Association with Foodborne Bacteria

Microorganisms ◽

10.3390/microorganisms8020201 ◽

2020 ◽

Vol 8 (2) ◽

pp. 201

Author(s):

Anja Klančnik ◽

Ivana Gobin ◽

Barbara Jeršek ◽

Sonja Smole Možina ◽

Darinka Vučković ◽

...

Keyword(s):

Escherichia Coli ◽

Listeria Monocytogenes ◽

Campylobacter Jejuni ◽

Biofilm Formation ◽

Risk Evaluation ◽

Acanthamoeba Castellanii ◽

Host Cells ◽

E Coli ◽

Adverse Conditions ◽

Ultrathin Sections

The aim of this study was to evaluate Campylobacter jejuni NTCT 11168 adhesion to abiotic and biotic surfaces when grown in co-culture with Escherichia coli ATCC 11229 and/or Listeria monocytogenes 4b. Adhesion of C. jejuni to polystyrene and to Caco-2 cells and Acanthamoeba castellanii was lower for at least 3 log CFU/mL compared to E. coli and L. monocytogenes. Electron micrographs of ultrathin sections revealed interactions of C. jejuni with host cells. In co-culture with E. coli and L. monocytogenes, adhesion of C. jejuni to all tested surfaces was significantly increased for more than 1 log CFU/mL. There was 10% higher aggregation for C. jejuni than for other pathogens, and high co-aggregation of co-cultures of C. jejuni with E. coli and L. monocytogenes. These data show that C. jejuni in co-cultures with E. coli and L. monocytogenes present significantly higher risk than C. jejuni as mono-cultures, which need to be taken into account in risk evaluation. C. jejuni adhesion is a prerequisite for their colonization, biofilm formation, and further contamination of the environment. C. jejuni survival under adverse conditions as a factor in their pathogenicity and depends on their adhesion to different surfaces, not only as individual strains, but also in co-cultures with other bacteria like E. coli and L. monocytogenes.

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