Lactobacillus plantarum Lipoteichoic Acids Possess Strain-Specific Regulatory Effects on the Biofilm Formation of Dental Pathogenic Bacteria

Bacterial biofilm residing in the oral cavity is closely related to the initiation and persistence of various dental diseases. Previously, we reported the anti-biofilm activity of Lactobacillus plantarum lipoteichoic acid (Lp.LTA) on a representative dental cariogenic pathogen, Streptococcus mutans. Since LTA structure varies in a bacterial strain-specific manner, LTAs from various L. plantarum strains may have differential anti-biofilm activity due to their distinct molecular structures. In the present study, we isolated Lp.LTAs from four different strains of L. plantarum (LRCC 5193, 5194, 5195, and 5310) and compared their anti-biofilm effects on the dental pathogens, including S. mutans, Enterococcus faecalis, and Streptococcus gordonii. All Lp.LTAs similarly inhibited E. faecalis biofilm formation in a dose-dependent manner. However, their effects on S. gordonii and S. mutans biofilm formation were different: LRCC 5310 Lp.LTA most effectively suppressed the biofilm formation of all strains of dental pathogens, while Lp.LTAs from LRCC 5193 and 5194 hardly inhibited or even enhanced the biofilm formation. Furthermore, LRCC 5310 Lp.LTA dramatically reduced the biofilm formation of the dental pathogens on the human dentin slice infection model. Collectively, these results suggest that Lp.LTAs have strain-specific regulatory effects on biofilm formation of dental pathogens and LRCC 5310 Lp.LTA can be used as an effective anti-biofilm agent for the prevention of dental infectious diseases.

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Antibiotics Application Strategies to Control Biofilm Formation in Pathogenic Bacteria

Current Pharmaceutical Biotechnology ◽

10.2174/1389201020666191112155905 ◽

2020 ◽

Vol 21 (4) ◽

pp. 270-286 ◽

Cited By ~ 2

Author(s):

Fazlurrahman Khan ◽

Dung T.N. Pham ◽

Sandra F. Oloketuyi ◽

Young-Mog Kim

Keyword(s):

Biofilm Formation ◽

Pathogenic Bacteria ◽

Extracellular Polymeric Substances ◽

Quorum Quenching ◽

Resistance Mechanisms ◽

Bacterial Biofilm ◽

Bacterial Cells ◽

High Concentration ◽

Biofilm Inhibition ◽

Abiotic Surfaces

Background: The establishment of a biofilm by most pathogenic bacteria has been known as one of the resistance mechanisms against antibiotics. A biofilm is a structural component where the bacterial community adheres to the biotic or abiotic surfaces by the help of Extracellular Polymeric Substances (EPS) produced by bacterial cells. The biofilm matrix possesses the ability to resist several adverse environmental factors, including the effect of antibiotics. Therefore, the resistance of bacterial biofilm-forming cells could be increased up to 1000 times than the planktonic cells, hence requiring a significantly high concentration of antibiotics for treatment. Methods: Up to the present, several methodologies employing antibiotics as an anti-biofilm, antivirulence or quorum quenching agent have been developed for biofilm inhibition and eradication of a pre-formed mature biofilm. Results: Among the anti-biofilm strategies being tested, the sub-minimal inhibitory concentration of several antibiotics either alone or in combination has been shown to inhibit biofilm formation and down-regulate the production of virulence factors. The combinatorial strategies include (1) combination of multiple antibiotics, (2) combination of antibiotics with non-antibiotic agents and (3) loading of antibiotics onto a carrier. Conclusion: The present review paper describes the role of several antibiotics as biofilm inhibitors and also the alternative strategies adopted for applications in eradicating and inhibiting the formation of biofilm by pathogenic bacteria.

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Application of Bacteriophages on Shiga Toxin-Producing Escherichia coli (STEC) Biofilm

Antibiotics ◽

10.3390/antibiotics10111423 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1423

Author(s):

Nicola Mangieri ◽

Roberto Foschino ◽

Claudia Picozzi

Keyword(s):

Escherichia Coli ◽

Shiga Toxin ◽

Biofilm Formation ◽

Pathogenic Bacteria ◽

Antimicrobial Agents ◽

Bacterial Cells ◽

Abiotic Surfaces ◽

Continuous Presence ◽

Different Strains ◽

Better Than

Shiga toxin-producing Escherichia coli are pathogenic bacteria able to form biofilms both on abiotic surfaces and on food, thus increasing risks for food consumers. Moreover, biofilms are difficult to remove and more resistant to antimicrobial agents compared to planktonic cells. Bacteriophages, natural predators of bacteria, can be used as an alternative to prevent biofilm formation or to remove pre-formed biofilm. In this work, four STEC able to produce biofilm were selected among 31 different strains and tested against single bacteriophages and two-phage cocktails. Results showed that our phages were able to reduce biofilm formation by 43.46% both when used as single phage preparation and as a cocktail formulation. Since one of the two cocktails had a slightly better performance, it was used to remove pre-existing biofilms. In this case, the phages were unable to destroy the biofilms and reduce the number of bacterial cells. Our data confirm that preventing biofilm formation in a food plant is better than trying to remove a preformed biofilm and the continuous presence of bacteriophages in the process environment could reduce the number of bacteria able to form biofilms and therefore improve the food safety.

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Effect of Lonicera caerulea var. emphyllocalyx Fruit on Biofilm Formed by Porphyromonas gingivalis

BioMed Research International ◽

10.1155/2019/3547858 ◽

2019 ◽

Vol 2019 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Masaaki Minami ◽

Hiroshi Takase ◽

Mineo Nakamura ◽

Toshiaki Makino

Keyword(s):

Oral Cavity ◽

Porphyromonas Gingivalis ◽

Aspiration Pneumonia ◽

Biofilm Formation ◽

Bacterial Colonization ◽

Bacterial Biofilm ◽

Dependent Manner ◽

Electron Microscopic ◽

Concentration Dependent Manner ◽

Lonicera Caerulea

Porphyromonas gingivalis is an important pathogenic anaerobic bacterium that causes aspiration pneumonia. This bacterium frequently forms biofilms in the oral cavity and in respiratory tract-associated medical devices. Bacterial colonization that occurs in association with this biofilm formation is the main reason for incurable aspiration pneumonia. The Lonicera caerulea var. emphyllocalyx (LCE) fruit has been used in folk medicine in Hokkaido, the northern part of Japan. The aim of this study was to elucidate one of the antimicrobial mechanisms of LCE methanol extract (LCEE)—the inhibitory effect of LCEE on biofilm formation by P. gingivalis. Our results show that LCEE significantly reduced biofilm formation by three different P. gingivalis isolates in a concentration- and time-dependent manner that were quantified by the adsorption of safranin red. When LCEE was added to biofilms already formed by P. gingivalis, LCEE did not degrade the biofilm. However, treatment with LCEE significantly promoted the removal of existing biofilm by vibration compared to that of control. We also confirmed biofilm formation in LCEE-treated P. gingivalis in tracheal tubes using scanning electron microscopic (SEM) analysis. Cyanidin 3-O-glucoside (C3G), one of the components of LCE, also inhibited the formation of biofilm by P. gingivalis in a concentration-dependent manner. Our results reveal that LCEE may be an effective antibacterial substance for P. gingivalis-induced aspiration pneumonia because of its role in the suppression of bacterial biofilm formation in the oral cavity.

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Gentamicin Combined With Hypoionic Shock Rapidly Eradicates Aquaculture Bacteria in vitro and in vivo

Frontiers in Microbiology ◽

10.3389/fmicb.2021.641846 ◽

2021 ◽

Vol 12 ◽

Author(s):

Yuanyuan Gao ◽

Zhongyu Chen ◽

Wei Yao ◽

Daliang Li ◽

Xinmiao Fu

Keyword(s):

Pathogenic Bacteria ◽

Bacterial Pathogens ◽

Combined Treatment ◽

Fish Farming ◽

Edwardsiella Tarda ◽

Small Scale ◽

Infection Model ◽

Dependent Manner ◽

Aquatic Animals

Bacterial pathogens are a major cause of infectious diseases in aquatic animals. The abuse of antibiotics in the aquatic industry has led to the proliferation of antibiotic resistance. It is therefore essential to develop more effective and safer strategies to increase the efficacy and extend the life span of the antibiotics used in aquaculture. In this study, we show that six aquaculture bacterial pathogens (i.e., Aeromonas hydrophila, Vibrio alginolyticus, Edwardsiella tarda, Streptococcus iniae, Vibrio harveyi, and Vibrio fluvialis) in the stationary phase can be rapidly killed after immersion in gentamicin- or neomycin-containing, ion-free solutions for a few minutes. Such hypoionic shock treatment enhances the bacterial uptake of gentamicin in an ATP-dependent manner. Importantly, we demonstrate, as a proof of concept, that gentamicin under hypoionic shock conditions can effectively kill A. hydrophila in vivo in a skin infection model of zebrafish (Danio rerio), completely curing the infected fish. Given that pathogenic bacteria generally adhere to the skin surface and gills of aquatic animals, our strategy is of potential significance for bacterial infection control, especially for small-scale economic fish farming and ornamental fish farming. Further, the combined treatment can be completed within 5 min with a relatively small volume of solution, thus minimizing the amount of residual antibiotics in both animals and the environment.

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Abstract 90: Serum Kawasaki Disease-specific Microbe-associated Molecular Patterns are Derived from in Vitro and in Vivo Biofilms

Circulation ◽

10.1161/circ.131.suppl_2.90 ◽

2015 ◽

Vol 131 (suppl_2) ◽

Author(s):

Takeshi Kusuda ◽

Yasutaka Nakashima ◽

Murata Kenji ◽

Shunsuke Kanno ◽

Mitsumasa Saito ◽

...

Keyword(s):

Biofilm Formation ◽

Pathogenic Bacteria ◽

Yersinia Pseudotuberculosis ◽

Molecular Structures ◽

Human Coronary Artery ◽

Medium Temperature ◽

Molecular Patterns ◽

Culture Supernatants

Background: We have found that serum KD-specific molecules possess molecular structures common to microbe-associated molecular patterns (MAMPs) from several bacteria by liquid chromatography-mass spectrometry (LC-MS) analysis. In the present study, we have extensively searched for the culture conditions that reproducibly produce MAMPs common to serum KD specific molecules. Methods: Yersinia pseudotuberculosis, spore-forming bacteria and pathogenic bacteria were obtained from KD patients. Others were purchased from American Type Culture Collection. Microbes were cultured in a variety of conditions (medium, temperature, duration, shaking and biofilm formation). For biofilm formation, a various kind of oil/butter was added to the culture medium. Extraction from in vitro samples (culture supernatants, bacteria cells, and biofilms) as well as in vivo biofilm samples (teeth, tongue, nasal cavity, or rectum) was performed with ethyl acetate. Samples were analyzed by HPLC and MS (Esquire 6000 electrospray ionization). Human coronary artery endothelial cells (HCAECs) were employed for cytokine assay. The concentrations of cytokines were measured by EC800 cell analyzer with a BD™ Cytometric Bead Array. Results: Serum KD-specific molecules showed m/z and MS/MS fragmentation patterns almost identical to those of MAMPs obtained from the biofilms formed in vitro (Bacillus cereus, Bacillus subtilis, Yersinia pseudotuberculosis, Staphylococcus aureus) or in vivo. Not culture supernatants but biofilm extracts from these bacteria, especially cultured in the presence of butter, induced proinflammatory cytokines by HCAECs. Conclusions: Extensive analysis by LC-MS/MS revealed that serum KD-specific molecules common to MAMPs were mostly derived from biofilms in vitro and in vivo. This report offers novel insight into the pathogenesis of KD.

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The phycobiota-derived proteins Pµ84 and Pµ19 suppress bacterial biofilm formation in gram-negative pathogens

10.5194/biofilms9-63 ◽

2020 ◽

Author(s):

Yuchen Han ◽

Wolfgang R. Streit ◽

Ines Krohn

Keyword(s):

Biofilm Formation ◽

Pathogenic Bacteria ◽

Bacterial Biofilm ◽

Klebsiella Pneumonia ◽

Biofilm Inhibition ◽

Marine Systems ◽

Microtiter Plates ◽

Small Proteins ◽

Ongoing Work ◽

Growth Promoting

Microalgae are typically found in freshwater and marine systems and they harbor a mostly a beneficial growth promoting microbiota. We have recently isolated several small proteins from the microbiomes of microalga (Scenedesmus quadricauda, Microasterias crux-melintensis, Chlorella saccherophilia) and have tested them for their role in either inhibition of biofilm formation and/or biofilm degradation. Thereby we have identified two candidate proteins that showed promising activities on biofilm inhibition and degradation. These proteins were designated P&#181;84 and P&#181;19 and strongly affected biofilm formation in several human- and plant-pathogenic bacteria. Recombinant and purified P&#181;84 and P&#181;19 were applied in biofilm assays in microtiter plates and reduced biofilms formed by Stenotrophomonas maltophilia, Pseudomonas aeruginosa and Klebsiella pneumonia. If expressed in the different hosts, biofilms were reduced by a factor of 40% and if applied as exogenous proteins, biofilms were reduced up to 20%. P&#181;84 application also resulted in a delayed biofilm formation and biofilm formation was affected by a factor of 17%. The microprotein P&#181;19 consist of 57 aa and P&#181;84 consists of 49 aa. Ongoing work elucidates the mechanism of P&#181;84 and P&#181;19 on the reduction of biofilm in order to achieve the optimal activity.

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In SilicoDiscovery andIn VitroValidation of Catechol-Containing Sulfonohydrazide Compounds as Potent Inhibitors of the Diguanylate Cyclase PleD

Journal of Bacteriology ◽

10.1128/jb.00742-15 ◽

2015 ◽

Vol 198 (1) ◽

pp. 147-156 ◽

Cited By ~ 14

Author(s):

Silvia Fernicola ◽

Alessandro Paiardini ◽

Giorgio Giardina ◽

Giordano Rampioni ◽

Livia Leoni ◽

...

Keyword(s):

Biofilm Formation ◽

Pathogenic Bacteria ◽

Caulobacter Crescentus ◽

Pharmacophore Model ◽

Second Messenger ◽

Binding Mode ◽

Structural Features ◽

Bacterial Biofilm ◽

Content Type

ABSTRACTBiofilm formation is responsible for increased antibiotic tolerance in pathogenic bacteria. Cyclic di-GMP (c-di-GMP) is a widely used second-messenger signal that plays a key role in bacterial biofilm formation. c-di-GMP is synthesized by diguanylate cyclases (DGCs), a conserved class of enzymes absent in mammals and hence considered attractive molecular targets for the development of antibiofilm agents. Here, the results of a virtual screening approach aimed at identifying small-molecule inhibitors of the DGC PleD fromCaulobacter crescentusare described. A three-dimensional (3D) pharmacophore model, derived from the mode of binding of GTP to the active site of PleD, was exploited to screen the ZINC database of compounds. Seven virtual hits were testedin vitrofor their ability to inhibit the activity of purified PleD by using circular dichroism spectroscopy. Two drug-like molecules with a catechol moiety and a sulfonohydrazide scaffold were shown to competitively inhibit PleD at the low-micromolar range (50% inhibitory concentration [IC50] of ∼11 μM). Their predicted binding mode highlighted key structural features presumably responsible for the efficient inhibition of PleD by both hits. These molecules represent the most potentin vitroinhibitors of PleD identified so far and could therefore result in useful leads for the development of novel classes of antimicrobials able to hamper biofilm formation.IMPORTANCEBiofilm-mediated infections are difficult to eradicate, posing a threatening health issue worldwide. The capability of bacteria to form biofilms is almost universally stimulated by the second messenger c-di-GMP. This evidence has boosted research in the last decade for the development of new antibiofilm strategies interfering with c-di-GMP metabolism. Here, two potent inhibitors of c-di-GMP synthesis have been identifiedin silicoand characterizedin vitroby using the well-characterized DGC enzyme PleD fromC. crescentusas a structural template and molecular target. Given that the protein residues implied as crucial for enzyme inhibition are found to be highly conserved among DGCs, the outcome of this study could pave the way for the future development of broad-spectrum antibiofilm compounds.

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Biosynthesis of zinc oxide nanoparticles by cell-biomass and supernatant of Lactobacillus plantarum TA4 and its antibacterial and biocompatibility properties

Scientific Reports ◽

10.1038/s41598-020-76402-w ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Hidayat Mohd Yusof ◽

Nor’Aini Abdul Rahman ◽

Rosfarizan Mohamad ◽

Uswatun Hasanah Zaidan ◽

Anjas Asmara Samsudin

Keyword(s):

Lactobacillus Plantarum ◽

Pathogenic Bacteria ◽

Biological Synthesis ◽

Hydroxyl Groups ◽

Dependent Manner ◽

Zno Nps ◽

Concentration Dependent Manner ◽

Stabilizing Agents ◽

Cell Biomass ◽

Vero Cell Line

AbstractThis study aims to utilize the cell-biomass (CB) and supernatant (CFS) of zinc-tolerant Lactobacillus plantarum TA4 as a prospective nanofactory to synthesize ZnO NPs. The surface plasmon resonance for the biosynthesized ZnO NPs-CFS and ZnO NPs-CB was 349 nm and 351 nm, respectively, thereby confirming the formation of ZnO NPs. The FTIR analysis revealed the presence of proteins, carboxyl, and hydroxyl groups on the surfaces of both the biosynthesized ZnO NPs that act as reducing and stabilizing agents. The DLS analysis revealed that the poly-dispersity indexes was less than 0.4 for both ZnO NPs. In addition, the HR-TEM micrographs of the biosynthesized ZnO NPs revealed a flower-like pattern for ZnO NPs-CFS and an irregular shape for ZnO NPs-CB with particles size of 291.1 and 191.8 nm, respectively. In this study, the biosynthesized ZnO NPs exhibited antibacterial activity against pathogenic bacteria in a concentration-dependent manner and showed biocompatibility with the Vero cell line at specific concentrations. Overall, CFS and CB of L. plantarum TA4 can potentially be used as a nanofactory for the biological synthesis of ZnO NPs.

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BhuR, a Virulence-Associated Outer Membrane Protein of Bordetella avium, Is Required for the Acquisition of Iron from Heme and Hemoproteins

Infection and Immunity ◽

10.1128/iai.70.10.5390-5403.2002 ◽

2002 ◽

Vol 70 (10) ◽

pp. 5390-5403 ◽

Cited By ~ 30

Author(s):

Erin R. Murphy ◽

Randy E. Sacco ◽

Amy Dickenson ◽

Daniel J. Metzger ◽

Yan Hu ◽

...

Keyword(s):

Outer Membrane ◽

Pathogenic Bacteria ◽

Whooping Cough ◽

Essential Element ◽

Local Environment ◽

Striking Similarity ◽

Infection Model ◽

Dependent Manner ◽

Upper Respiratory Tract ◽

Bordetella Avium

ABSTRACT Iron (Fe) is an essential element for most organisms which must be obtained from the local environment. In the case of pathogenic bacteria, this fundamental element must be acquired from the fluids and tissues of the infected host. A variety of systems have evolved in bacteria for efficient acquisition of host-bound Fe. The gram-negative bacterium Bordetella avium, upon colonization of the avian upper respiratory tract, produces a disease in birds that has striking similarity to whooping cough, a disease caused by the obligate human pathogen Bordetella pertussis. We describe a B. avium Fe utilization locus comprised of bhuR and six accessory genes (rhuIR and bhuSTUV). Genetic manipulations of B. avium confirmed that bhuR, which encodes a putative outer membrane heme receptor, mediates efficient acquisition of Fe from hemin and hemoproteins (hemoglobin, myoglobin, and catalase). BhuR contains motifs which are common to bacterial heme receptors, including a consensus FRAP domain, an NPNL domain, and two TonB boxes. An N-terminal 32-amino-acid segment, putatively required for rhuIR-dependent regulated expression of bhuR, is present in BhuR but not in other bacterial heme receptors. Two forms of BhuR were observed in the outer membrane of B. avium: a 91-kDa polypeptide consistent in size with the predicted mature protein and a smaller 82-kDa polypeptide which lacks the 104 amino acids found at the N terminus of the 91-kDa form. A mutation in hemA was engineered in B. avium to demonstrate that the bacterium transports heme into the cytoplasm in a BhuR-dependent manner. The role of BhuR in virulence was established in turkey poults by use of a competitive-infection model.

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