scholarly journals Type IV Pili and the CcpA Protein Are Needed for Maximal Biofilm Formation by the Gram-Positive Anaerobic Pathogen Clostridium perfringens

2008 ◽  
Vol 76 (11) ◽  
pp. 4944-4951 ◽  
Author(s):  
John J. Varga ◽  
Blair Therit ◽  
Stephen B. Melville
Keyword(s):  
Environmental Stress ◽  
Clostridium Perfringens ◽  
Biofilm Formation ◽  
Atmospheric Oxygen ◽  
Gliding Motility ◽  
Penicillin G ◽  
Gram Positive ◽  
Planktonic Growth ◽  
Type Iv ◽  
Biofilm Development

ABSTRACT The predominant organizational state of bacteria in nature is biofilms. Biofilms have been shown to increase bacterial resistance to a variety of stresses. We demonstrate for the first time that the anaerobic gram-positive pathogen Clostridium perfringens forms biofilms. At the same concentration of glucose in the medium, optimal biofilm formation depended on a functional CcpA protein. While the ratio of biofilm to planktonic growth was higher in the wild type than in a ccpA mutant strain in middle to late stages of biofilm development, the bacteria shifted from a predominantly biofilm state to planktonic growth as the concentration of glucose in the medium increased in a CcpA-independent manner. As is the case in some gram-negative bacteria, type IV pilus (TFP)-dependent gliding motility was necessary for efficient biofilm formation, as demonstrated by laser confocal and electron microscopy. However, TFP were not associated with the bacteria in the biofilm but with the extracellular matrix. Biofilms afforded C. perfringens protection from environmental stress, including exposure to atmospheric oxygen for 6 h and 24 h and to 10 mM H2O2 for 5 min. Biofilm cells also showed 5- to 15-fold-increased survival over planktonic cells after exposure to 20 μg/ml (27 times the MIC) of penicillin G for 6 h and 24 h, respectively. These results indicate C. perfringens biofilms play an important role in the persistence of the bacteria in response to environmental stress and that they may be a factor in diseases, such as antibiotic-associated diarrhea and gas gangrene, that are caused by C. perfringens.

Tectorigenin reduces type IV pilus-dependent cell adherence in Clostridium perfringens

2019 ◽  
Vol 366 (10) ◽  
Author(s):  
Shui Liu ◽  
Jianfeng Wang ◽  
Jian Zhang ◽  
Tingting Wang ◽  
Yonglin Zhou ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Clostridium Perfringens ◽  
Biofilm Formation ◽  
Chinese Herb ◽  
Inhibitory Effect ◽  
Gliding Motility ◽  
Host Cells ◽  
Type Iv Pilus ◽  
Type Iv ◽  
Dependent Cell

Abstract Clostridium perfringens is an anaerobic, Gram-positive bacterium that causes a range of diseases in humans and animals around the globe. The type IV pilus (TFP) system plays a key role in the colonization and invasion of host cells, biofilm formation and gliding motility, which is vital for C. perfringens infection. Therefore, targeting TFP function may be a promising strategy for the treatment of C. perfringens infection. Here, we investigated the potential inhibitory effects of tectorigenin (TE), an isoflavone extracted from the rhizome of the Chinese herb Belamcanda chinensis (L.) DC, on gliding motility, biofilm formation, adherence to cells and antibacterial activity of C. perfringens. Tectorigenin significantly inhibited gliding motility, biofilm formation and adherence to Caco-2 cells without observable antibacterial activity against C. perfringens. In addition, we also demonstrated that the inhibitory effect of TE on TFP function appears to be partially achieved by the suppression of TFP-associated genes. These findings demonstrate that TE may have the potential to be developed as a new anti-virulence drug for C. perfringens infection, particularly for the targeting of TFP.

Type IV pili‐dependent gliding motility in the Gram‐positive pathogenClostridium perfringensand other Clostridia

2006 ◽  
Vol 62 (3) ◽  
pp. 680-694 ◽  
Author(s):  
John J. Varga ◽  
Van Nguyen ◽  
David K. O'Brien ◽  
Katherine Rodgers ◽  
Richard A. Walker ◽  
...  
Keyword(s):  
Type Iv Pili ◽  
Gliding Motility ◽  
Gram Positive ◽  
Type Iv

scholarly journals Cyclic Di-GMP Binding by an Assembly ATPase (PilB2) and Control of Type IV Pilin Polymerization in the Gram-Positive Pathogen Clostridium perfringens

2017 ◽  
Vol 199 (10) ◽  
Author(s):  
William A. Hendrick ◽  
Mona W. Orr ◽  
Samantha R. Murray ◽  
Vincent T. Lee ◽  
Stephen B. Melville
Keyword(s):  
Clostridium Perfringens ◽  
Biochemical Characterization ◽  
Core Protein ◽  
Type Iv Pili ◽  
Host Cells ◽  
Dependent Manner ◽  
Gram Positive ◽  
Content Type ◽  
Type Iv

ABSTRACT The Gram-positive pathogen Clostridium perfringens possesses type IV pili (TFP), which are extracellular fibers that are polymerized from a pool of pilin monomers in the cytoplasmic membrane. Two proteins that are essential for pilus functions are an assembly ATPase (PilB) and an inner membrane core protein (PilC). Two homologues each of PilB and PilC are present in C. perfringens, called PilB1/PilB2 and PilC1/PilC2, respectively, along with four pilin proteins, PilA1 to PilA4. The gene encoding PilA2, which is considered the major pilin based on previous studies, is immediately downstream of the pilB2 and pilC2 genes. Purified PilB2 had ATPase activity, bound zinc, formed hexamers even in the absence of ATP, and bound the second messenger molecule cyclic di-GMP (c-di-GMP). Circular dichroism spectroscopy of purified PilC2 indicated that it retained its predicted degree of alpha-helical secondary structure. Even though no direct interactions between PilB2 and PilC2 could be detected in vivo or in vitro even in the presence of c-di-GMP, high levels of expression of a diguanylate cyclase from C. perfringens (CPE1788) stimulated polymerization of PilA2 in a PilB2- and PilC2-dependent manner. These results suggest that PilB2 activity is controlled by c-di-GMP levels in vivo but that PilB2-PilC2 interactions are either transitory or of low affinity, in contrast to results reported previously from in vivo studies of the PilB1/PilC1 pair in which PilC1 was needed for polar localization of PilB1. This is the first biochemical characterization of a c-di-GMP-dependent assembly ATPase from a Gram-positive bacterium. IMPORTANCE Type IV pili (TFP) are protein fibers involved in important bacterial functions, including motility, adherence to surfaces and host cells, and natural transformation. All clostridia whose genomes have been sequenced show evidence of the presence of TFP. The genetically tractable species Clostridium perfringens was used to study proteins involved in polymerizing the pilin, PilA2, into a pilus. The assembly ATPase PilB2 and its cognate membrane protein partner, PilC2, were purified. PilB2 bound the intracellular signal molecule c-di-GMP. Increased levels of intracellular c-di-GMP led to increased polymerization of PilA2, indicating that Gram-positive bacteria use this molecule to regulate pilus synthesis. These findings provide valuable information for understanding how pathogenic clostridia regulate TFP to cause human diseases.

scholarly journals Xanthomonas citri subsp. citri Type IV Pilus Is Required for Twitching Motility, Biofilm Development, and Adherence

2014 ◽  
Vol 27 (10) ◽  
pp. 1132-1147 ◽  
Author(s):  
German Dunger ◽  
Cristiane R. Guzzo ◽  
Maxuel O. Andrade ◽  
Jeffrey B. Jones ◽  
Chuck S. Farah
Keyword(s):  
Biofilm Formation ◽  
Defense Mechanisms ◽  
Large Set ◽  
Twitching Motility ◽  
Xanthomonas Citri ◽  
Helical Polymers ◽  
Type Iv ◽  
Bacterial Migration ◽  
In The Wild ◽  
Biofilm Development

Bacterial type IV pili (T4P) are long, flexible surface filaments that consist of helical polymers of mostly pilin subunits. Cycles of polymerization, attachment, and depolymerization mediate several pilus-dependent bacterial behaviors, including twitching motility, surface adhesion, pathogenicity, natural transformation, escape from immune system defense mechanisms, and biofilm formation. The Xanthomonas citri subsp. citri strain 306 genome codes for a large set of genes involved in T4P biogenesis and regulation and includes several pilin homologs. We show that X. citri subsp. citri can exhibit twitching motility in a manner similar to that observed in other bacteria such as Pseudomonas aeruginosa and Xylella fastidiosa and that this motility is abolished in Xanthomonas citri subsp. citri knockout strains in the genes coding for the major pilin subunit PilAXAC3241, the ATPases PilBXAC3239 and PilTXAC2924, and the T4P biogenesis regulators PilZXAC1133 and FimXXAC2398. Microscopy analyses were performed to compare patterns of bacterial migration in the wild-type and knockout strains and we observed that the formation of mushroom-like structures in X. citri subsp. citri biofilm requires a functional T4P. Finally, infection of X. citri subsp. citri cells by the bacteriophage (ΦXacm4-11 is T4P dependent. The results of this study improve our understanding of how T4P influence Xanthomonas motility, biofilm formation, and susceptibility to phage infection.

scholarly journals Tooth mousse containing casein phosphopeptide-amorphous calcium phosphate prevents biofilm formation of Streptococcus mutans

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ronit Vogt Sionov ◽  
Danae Tsavdaridou ◽  
Muna Aqawi ◽  
Batya Zaks ◽  
Doron Steinberg ◽  
...  
Keyword(s):  
Calcium Phosphate ◽  
Streptococcus Mutans ◽  
Amorphous Calcium Phosphate ◽  
Biofilm Formation ◽  
Laser Scanning Microscopy ◽  
Planktonic Growth ◽  
Casein Phosphopeptide ◽  
Cariogenic Bacterium ◽  
Biofilm Development ◽  
Tooth Mousse

Abstract Background Streptococcus mutans is a common cariogenic bacterium in the oral cavity involved in plaque formation. Casein phosphopeptide-amorphous calcium phosphate (CPP-ACP) has been introduced into tooth mousse to encourage remineralization of dental enamel. The aim of this research was to study the effect of tooth mousse containing CPP-ACP (GC Tooth Mousse®) or CPP-ACP with 0.2% fluoride (CPP-ACPF; GC Tooth Mousse Plus®; GCP) on S. mutans planktonic growth and biofilm formation. Methods S. mutans was cultivated in the presence of different dilutions of the tooth mousse containing CPP-ACP or CPP-ACPF, and the planktonic growth was determined by ATP viability assay and counting colony-forming units (CFUs). The resulting biofilms were examined by crystal violet staining, MTT metabolic assay, confocal laser scanning microscopy (CLSM), and scanning electron microscope (SEM). Results The CPP-ACP tooth mousse (GC) at a dilution of 5–50 mg/ml (0.5–5%) did not inhibit planktonic growth, and even increased the ATP content and the number of viable bacteria after a 24 h incubation. The same was observed for the CPP-ACPF tooth mousse (GCP), except for the higher concentrations (25 and 50 mg/ml) that led to a drop in the bacterial count. Importantly, both compounds significantly decreased S. mutans biofilm formation at dilutions as low as 1.5–3 mg/ml. 12.5 mg/ml GC and 6.25 mg/ml GCP inhibited biofilm formation by 90% after 4 h. After 24 h, the MBIC90 was 6.25 mg/ml for both. CLSM images confirmed the strong inhibitory effect GC and GCP had on biofilm formation when using 5 mg/ml tooth mousse. SEM images of those bacteria that managed to form biofilm in the presence of 5 mg/ml tooth mousse, showed alterations in the bacterial morphology, where the streptococci appear 25–30% shorter on the average than the control bacteria. Conclusion Our data show that the tooth mousse containing CPP-ACP reduces biofilm formation of the cariogenic bacterium S. mutans without killing the bacteria. The use of natural substances which inhibit biofilm development without killing the bacteria, has therapeutic benefits, especially in orthodontic pediatric patients.

scholarly journals Loss of Flagellum-Based Motility by Listeria monocytogenes Results in Formation of Hyperbiofilms

2008 ◽  
Vol 190 (17) ◽  
pp. 6030-6034 ◽  
Author(s):  
Tatsaporn Todhanakasem ◽  
Glenn M. Young
Keyword(s):  
Listeria Monocytogenes ◽  
Biofilm Formation ◽  
High Density ◽  
Gram Positive ◽  
Surface Attachment ◽  
Bacterial Surface ◽  
Food Borne ◽  
Biofilm Development

ABSTRACT Biofilm formation by the gram-positive, motile, food-borne pathogen Listeria monocytogenes was demonstrated to occur by an ordered series of stages. Biofilm development involves flagellum-based motility, which when blocked decreases initial bacterial surface attachment but subsequently leads to the formation of hyperbiofilms, surface-attached communities reaching high density.

Efficacy of metal ions and isothiazolones in inhibiting Enterobacter cloacae BF-17 biofilm formation

2014 ◽  
Vol 60 (1) ◽  
pp. 5-14 ◽  
Author(s):  
Gang Zhou ◽  
Long-jie Li ◽  
Qing-shan Shi ◽  
You-sheng Ouyang ◽  
Yi-ben Chen ◽  
...  
Keyword(s):  
Metal Ions ◽  
Biofilm Formation ◽  
High Capacity ◽  
Enterobacter Cloacae ◽  
High Concentration ◽  
Planktonic Growth ◽  
Low Concentrations ◽  
Negative Effect ◽  
Biofilm Development ◽  
Initial Biofilm

Enterobacter cloacae is a nosocomial pathogen. The E. cloacae strain BF-17, with a high capacity for biofilm formation, was screened and identified from industrially contaminated samples, carried out in our laboratory. To develop an efficient strategy to deal with biofilms, we investigated the effects of metal ions, including Na+, K+, Ca2+, Mg2+, Cu2+, and Mn2+, and 3 isothiazolones, on elimination of E. cloacae BF-17 biofilm formation by using a 0.1% crystal violet staining method. The results revealed that higher concentrations of Na+ or K+ significantly inhibited E. cloacae BF-17 biofilm development. Meanwhile, Ca2+ and Mn2+ stimulated biofilm formation at low concentration but exhibited a negative effect at high concentration. Moreover, biofilm formation decreased with increasing concentration of Mg2+ and Cu2+. The isothiazolones Kathon (14%), 1,2-benzisothiazolin-3-one (11%), and 2-methyl-4-isothiazolin-3-one (10%) stimulated initial biofilm formation but not planktonic growth at low concentrations and displayed inhibitory effects on both biofilm formation and planktonic growth at higher concentrations. Unfortunately, the 3 isothiazolones exerted negligible effects on preformed or fully mature biofilms. Our findings suggest that Na+, K+, Mg2+, and isothiazolones could be used to prevent and eliminate E. cloacae BF-17 biofilms.

scholarly journals Initial Phases of Biofilm Formation in Shewanella oneidensis MR-1

2004 ◽  
Vol 186 (23) ◽  
pp. 8096-8104 ◽  
Author(s):  
Kai M. Thormann ◽  
Renée M. Saville ◽  
Soni Shukla ◽  
Dale A. Pelletier ◽  
Alfred M. Spormann
Keyword(s):  
Biofilm Formation ◽  
Three Dimensional ◽  
Shewanella Oneidensis ◽  
Lateral Spreading ◽  
Flow Chamber ◽  
Natural Environments ◽  
Initial Attachment ◽  
Mn Oxide ◽  
Type Iv ◽  
Biofilm Development

ABSTRACT Shewanella oneidensis MR-1 is a facultative Fe(III)- and Mn(IV)-reducing microorganism and serves as a model for studying microbially induced dissolution of Fe or Mn oxide minerals as well as biogeochemical cycles. In soil and sediment environments, S. oneidensis biofilms form on mineral surfaces and are critical for mediating the metabolic interaction between this microbe and insoluble metal oxide phases. In order to develop an understanding of the molecular basis of biofilm formation, we investigated S. oneidensis biofilms developing on glass surfaces in a hydrodynamic flow chamber system. After initial attachment, growth of microcolonies and lateral spreading of biofilm cells on the surface occurred simultaneously within the first 24 h. Once surface coverage was almost complete, biofilm development proceeded with extensive vertical growth, resulting in formation of towering structures giving rise to pronounced three-dimensional architecture. Biofilm development was found to be dependent on the nutrient conditions, suggesting a metabolic control. In global transposon mutagenesis, 173 insertion mutants out of 15,000 mutants screened were identified carrying defects in initial attachment and/or early stages in biofilm formation. Seventy-one of those mutants exhibited a nonswimming phenotype, suggesting a role of swimming motility or motility elements in biofilm formation. Disruption mutations in motility genes (flhB, fliK, and pomA), however, did not affect initial attachment but affected progression of biofilm development into pronounced three-dimensional architecture. In contrast, mutants defective in mannose-sensitive hemagglutinin type IV pilus biosynthesis and in pilus retraction (pilT) showed severe defects in adhesion to abiotic surfaces and biofilm formation, respectively. The results provide a basis for understanding microbe-mineral interactions in natural environments.

scholarly journals Expression of Xylella fastidiosa Fimbrial and Afimbrial Proteins during Biofilm Formation

2010 ◽  
Vol 76 (13) ◽  
pp. 4250-4259 ◽  
Author(s):  
R. Caserta ◽  
M. A. Takita ◽  
M. L. Targon ◽  
L. K. Rosselli-Murai ◽  
A. P. de Souza ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Polyclonal Antibodies ◽  
Xylella Fastidiosa ◽  
Immunogold Electron Microscopy ◽  
Insect Vector ◽  
Plant Host ◽  
Type Iv ◽  
Genes Encoding ◽  
Complete Sequencing ◽  
Biofilm Development

ABSTRACT Complete sequencing of the Xylella fastidiosa genome revealed characteristics that have not been described previously for a phytopathogen. One characteristic of this genome was the abundance of genes encoding proteins with adhesion functions related to biofilm formation, an essential step for colonization of a plant host or an insect vector. We examined four of the proteins belonging to this class encoded by genes in the genome of X. fastidiosa: the PilA2 and PilC fimbrial proteins, which are components of the type IV pili, and XadA1 and XadA2, which are afimbrial adhesins. Polyclonal antibodies were raised against these four proteins, and their behavior during biofilm development was assessed by Western blotting and immunofluorescence assays. In addition, immunogold electron microscopy was used to detect these proteins in bacteria present in xylem vessels of three different hosts (citrus, periwinkle, and hibiscus). We verified that these proteins are present in X. fastidiosa biofilms but have differential regulation since the amounts varied temporally during biofilm formation, as well as spatially within the biofilms. The proteins were also detected in bacteria colonizing the xylem vessels of infected plants.

Comparative and Correlative Analysis of Biofilm Formation and Antimicrobial Resistance Traits of Pathogenic Bacterial Isolates

2021 ◽  
pp. 71-91
Author(s):  
Asif Shahriar ◽  
Kamal Kanta Das ◽  
Tamanna Islam ◽  
Farahnaaz Feroz
Keyword(s):  
Biofilm Formation ◽  
Pathogenic Bacteria ◽  
Opportunistic Infections ◽  
Penicillin G ◽  
P Value ◽  
Drug Resistant ◽  
Bacterial Isolates ◽  
Gram Positive ◽  
Beta Lactamases ◽  
Recent Emergence

Background: Pathogenic bacteria mostly cause emerging infectious diseases, including community-acquired, nosocomial, and opportunistic infections. The recent emergence of biofilms and the production of beta-lactamases strains have led to the widespread number of drug-resistant bacterial pathogens associated with infectious illness and death. Materials and Methods: This study was carried out to detect the ability of biofilm formation by gram-negative and gram-positive pathogenic isolates collected from clinical environments and determine their multi-drug resistant traits along with the detection of Extended-spectrum beta-lactamases (ESBLs) and Metallo-beta-lactamases (MBLs) production. Results: Among the 15 pathogenic isolates, A. baumannii (85%) were ESBL positive and (65%) MBL positive; P. mirabilis was ESBL and MBL positive (65% and 90%), respectively. The uropathogenic bacterial isolates, both K. pneumoniae and E. coli were found to be ESBL positive (60%) and MBL positive (40%); ESBL positive (75%) and MBL positive (60%), respectively. Whereas the pathogenic P. aeruginosa isolates were found to be ESBL and MBL positive (100%). In contrast, among the three gram-positive S. aureus isolates, two isolates were found to be ESBL positive (85%) and one isolate was found to be MBL positive (35%). The association between ESBL and MBL production and biofilm formation was statistically significant (P value=0.001). Besides, most active drugs, penicillin G, kanamycin, ciprofloxacin, ceftazidime, azithromycin and erythromycin, were resistant against all pathogenic isolates, whereas pathogens were significantly sensitive against Cotrimoxazole, Erythromycin, Streptomycin, and Ceftriaxone. Conclusion: This experiment's findings indicate the emergence and rapid spread of such multidrug-resistant pathogens are of great concern. Early detection of ESBL and MBL-producing pathogens is of paramount clinical significance; therefore, strict infection control practices and therapeutic guidance for confirmed infection can be rapidly initiated.

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