Genetic analysis of Treponema denticola ATCC 35405 biofilm formation

Microbiology ◽  
2004 ◽  
Vol 150 (7) ◽  
pp. 2401-2407 ◽  
Author(s):  
Peter M. Vesey ◽  
Howard K. Kuramitsu
Keyword(s):  
Genetic Analysis ◽  
Periodontal Disease ◽  
Biofilm Formation ◽  
Oral Bacteria ◽  
Treponema Denticola ◽  
Biofilm Model ◽  
Outer Sheath ◽  
Specific Protease ◽  
Cytoplasmic Filament ◽  
Inert Surfaces

Treponema denticola is a major aetiological organism implicated in periodontal disease. The interaction of T. denticola with other oral bacteria, in particular Porphyromonas gingivalis, in biofilm formation is thought to be an important step in the onset of periodontal disease. The interaction between T. denticola and P. gingivalis has been examined using a panel of T. denticola mutants and their effects on mixed biofilm formation tested in a static biofilm model. T. denticola ATCC 35405 did not form detectable biofilms on various inert surfaces. However, the spirochaete was demonstrated to form a biofilm with preattached P. gingivalis 381. T. denticola cfpA, which lacks the cytoplasmic filament, was unable to produce a mixed biofilm with P. gingivalis. A T. denticola flgE mutant which lacks the flagella hook protein and is therefore non-motile displayed a reduced, but readily detectable, ability to form a mixed biofilm as did the T. denticola mutant which does not possess the major outer sheath protein (Msp). The T. denticola lrrA mutant was only moderately defective in forming mixed biofilms with P. gingivalis. However, the T. denticola methyl-accepting chemotaxis protein (DmcA) did not appear to play a major role in mixed biofilm formation. In contrast, T. denticola lacking the PrtP protein for prolyl-phenylalanine-specific protease, showed an increased ability to form mixed biofilms and a prolonged viability in the biofilm.

scholarly journals The Antibacterial Activity of LL-37 against Treponema denticola Is Dentilisin Protease Independent and Facilitated by the Major Outer Sheath Protein Virulence Factor

2011 ◽  
Vol 80 (3) ◽  
pp. 1107-1114 ◽  
Author(s):  
Graciela Rosen ◽  
Michael N. Sela ◽  
Gilad Bachrach
Keyword(s):  
Periodontal Disease ◽  
Host Defense ◽  
Surface Protein ◽  
Human Saliva ◽  
Treponema Denticola ◽  
Host Defense Peptides ◽  
Content Type ◽  
Outer Sheath ◽  
Major Surface

Host defense peptides are innate immune effectors that possess both bactericidal activities and immunomodulatory functions. Deficiency in the human host defense peptide LL-37 has previously been correlated with severe periodontal disease.Treponema denticolais an oral anaerobic spirochete closely associated with the pathogenesis of periodontal disease. TheT. denticolamajor surface protein (MSP), involved in adhesion and cytotoxicity, and the dentilisin serine protease are key virulence factors of this organism. In this study, we examined the interactions between LL-37 andT. denticola. The threeT. denticolastrains tested were susceptible to LL-37. Dentilisin was found to inactivate LL-37 by cleaving it at the Lys, Phe, Gln, and Val residues. However, dentilisin deletion did not increase the susceptibility ofT. denticolato LL-37. Furthermore, dentilisin activity was found to be inhibited by human saliva. In contrast, a deficiency of theT. denticolaMSP increased resistance to LL-37. The MSP-deficient mutant bound less fluorescently labeled LL-37 than the wild-type strain. MSP demonstrated specific, dose-dependent LL-37 binding. In conclusion, though capable of LL-37 inactivation, dentilisin does not protectT. denticolafrom LL-37. Rather, the rapid, MSP-mediated binding of LL-37 to the treponemal outer sheath precedes cleavage by dentilisin. Moreover,in vivo, saliva inhibits dentilisin, thus preventing LL-37 restriction and ensuring its bactericidal and immunoregulatory activities.

scholarly journals The Msp Protein of Treponema denticola Interrupts Activity of Phosphoinositide Processing in Neutrophils

2019 ◽  
Vol 87 (11) ◽  
Author(s):  
Megan M. Jones ◽  
Stephen T. Vanyo ◽  
Michelle B. Visser
Keyword(s):  
Immune Response ◽  
Periodontal Disease ◽  
Alveolar Bone ◽  
Membrane Vesicles ◽  
Neutrophil Function ◽  
Oral Bacteria ◽  
Outer Membrane Vesicles ◽  
Treponema Denticola ◽  
Content Type ◽  
Tensin Homolog

ABSTRACT Periodontal disease is a significant health burden, causing tooth loss and poor oral and overall systemic health. Dysbiosis of the oral biofilm and a dysfunctional immune response drive chronic inflammation, causing destruction of soft tissue and alveolar bone supporting the teeth. Treponema denticola, a spirochete abundant in the plaque biofilm of patients with severe periodontal disease, perturbs neutrophil function by modulating appropriate phosphoinositide (PIP) signaling. Through a series of immunoblotting and quantitative PCR (qPCR) experiments, we show that Msp does not alter the gene transcription or protein content of key enzymes responsible for PIP3 signaling: 3′ phosphatase and tensin homolog (PTEN), phosphatidylinositol 3-kinase (PI3K), or 5′ Src homology 2 domain-containing inositol phosphatase 1 (SHIP1). Instead, using immunoblotting and enzyme-linked immunosorbent assays (ELISAs), we found that Msp activates PTEN through dephosphorylation specifically at the S380 site. Msp in intact organisms or outer membrane vesicles also restricts PIP signaling. SHIP1 phosphatase release was assessed using chemical inhibition and immunoprecipitation to show that Msp moderately decreases SHIP1 activity. Msp also prevents secondary activation of the PTEN/PI3K response. We speculate that this result is due to the redirection of the PIP3 substrate away from SHIP1 to PTEN. Immunofluorescence microscopy revealed a redistribution of PTEN from the cytoplasm to the plasma membrane following exposure to Msp, which may contribute to PTEN activation. Mechanisms of how T. denticola modulates and evades the host immune response are still poorly described, and here we provide further mechanistic evidence of how spirochetes modify PIP signaling to dampen neutrophil function. Understanding how oral bacteria evade the immune response to perpetuate the cycle of inflammation and infection is critical for combating periodontal disease to improve overall health outcomes.

scholarly journals A polymicrobial biofilm model for testing the antimicrobial potential of a nisin-biogel for canine periodontal disease control

2020 ◽  
Vol 16 (1) ◽  
Author(s):  
Eva Cunha ◽  
Sandra Rebelo ◽  
Carla Carneiro ◽  
Luís Tavares ◽  
Luís Miguel Carreira ◽  
...  
Keyword(s):  
Periodontal Disease ◽  
Guar Gum ◽  
Bacterial Species ◽  
Density Measurement ◽  
Oral Bacteria ◽  
Tooth Surface ◽  
Pd Control ◽  
Biofilm Model

Abstract Background Periodontal disease (PD) in dogs is prompted by the establishment of a polymicrobial biofilm at the tooth surface and a subsequent host inflammatory response. Several strategies may be used for PD control, including dental hygiene home care procedures, like toothbrushing, special diet and chew toys that reduce dental plaque accumulation, or professional periodontal treatments. Aiming at PD control, a biogel composed by nisin and guar-gum was previously developed. This work aimed to establish an in vitro model mimicking the PD-associated biofilms and to evaluate the nisin-biogel inhibitory activity against this polymicrobial biofilm by determining its Minimum Biofilm Inhibitory (MBIC) and Eradication Concentrations (MBEC). Bacterial species tested included Neisseria zoodegmatis CCUG 52598T, Corynebacterium canis CCUG 58627T, Porphyromonas cangingivalis DSMZ VPB 4874, Peptostreptococcus canis CCUG 57081 and an Enterococcus faecalis isolate belonging to a collection of oral bacteria obtained from dogs with PD. Before establishing the biofilm, coaggregation between species was determined by optical density measurement after 2 and 24 hours. Nisin-biogel MBIC and MBEC values regarding the polymicrobial biofilm were determined using a modified version of the Calgary biofilm pin lid device, after confirming the presence of the five bacterial species by Fluorescent In Situ Hybridization. Results Only 40% of the bacterial dual suspensions were able to coaggregate at 2 hours, but all species tested exhibited a coaggregation percentage higher than 30% at 24 hours. It was possible to establish a 48 h polymicrobial biofilm model composed by the five bacterial species selected. This model was used to determine nisin-biogel MBIC (26.39 ± 5.89 µg/mL) and MBEC (62.5 ± 27.73 µg/mL) values. Conclusions Our results showed that the nisin-biogel can inhibit and eradicate PD multispecies biofilms. As this in vitro model mimics an in vivo periodontal polymicrobial biofilm, our results reinforce the potential of the application of nisin-biogel for canine PD control.

Comparison of Sampling Techniques For qPCR Quantification of Periodontal Pathogens

2018 ◽  
Vol 68 (12) ◽  
pp. 2853-2856 ◽  
Author(s):  
Igor Jelihovschi ◽  
Cristian Drochioi ◽  
Aida Corina Badescu ◽  
Raoul Vasile Lupusoru ◽  
Alexandra Elena Munteanu ◽  
...  
Keyword(s):  
Periodontal Disease ◽  
Periodontal Diseases ◽  
Pcr Analysis ◽  
Treponema Denticola ◽  
Subgingival Plaque ◽  
Periodontal Pathogens ◽  
Radiographic Evidence ◽  
Gingival Bleeding ◽  
As Species ◽  
Qpcr Quantification

The diagnosis of periodontal disease is mainly based on use of clinical and radiographic evidence. In this study we employed a quantitative PCR analysis of Aggregatibacter actinomycetemcomitans and Treponema denticola as species strongly involved in periodontal diseases, burden in periodontal pockets to detect the main sampling factors that interfere with qPCR results. From 22 patients with advanced periodontal disease, subgingival plaque was comparatively collected by paper points and periodontal Gracey curettes. Samples were collected from the same situs in presence of gingival bleeding and absence of bleeding. The concordance and agreement of results between samples were assessed. The present study demonstrates that subgingival plaque sampling with sterile absorbable paper points is often accompanied by gingival bleeding resulting in quantification biases of periodontal pathogens.

scholarly journals In Vitro Anti-Biofilm and Antibacterial Properties of Streptococcus downii sp. nov.

2021 ◽  
Vol 9 (2) ◽  
pp. 450
Author(s):  
Maigualida Cuenca ◽  
María Carmen Sánchez ◽  
Pedro Diz ◽  
Lucía Martínez-Lamas ◽  
Maximiliano Álvarez ◽  
...  
Keyword(s):  
Antibacterial Activity ◽  
Quantitative Polymerase Chain Reaction ◽  
Bacterial Load ◽  
Antibacterial Properties ◽  
Antibacterial Activities ◽  
Oral Bacteria ◽  
Periodontal Pathogens ◽  
Biofilm Model ◽  
Biofilm Development

The aim of this study was to evaluate the potential anti-biofilm and antibacterial activities of Streptococcus downii sp. nov. To test anti-biofilm properties, Streptococcus mutans, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans were grown in a biofilm model in the presence or not of S. downii sp. nov. for up to 120 h. For the potential antibacterial activity, 24 h-biofilms were exposed to S. downii sp. nov for 24 and 48 h. Biofilms structures and bacterial viability were studied by microscopy, and the effect in bacterial load by quantitative polymerase chain reaction. A generalized linear model was constructed, and results were considered as statistically significant at p < 0.05. The presence of S. downii sp. nov. during biofilm development did not affect the structure of the community, but an anti-biofilm effect against S. mutans was observed (p < 0.001, after 96 and 120 h). For antibacterial activity, after 24 h of exposure to S. downii sp. nov., counts of S. mutans (p = 0.019) and A. actinomycetemcomitans (p = 0.020) were significantly reduced in well-structured biofilms. Although moderate, anti-biofilm and antibacterial activities of S. downii sp. nov. against oral bacteria, including some periodontal pathogens, were demonstrated in an in vitro biofilm model.

scholarly journals Blocking of Bacterial Biofilm Formation by a Fish Protein Coating

2008 ◽  
Vol 74 (11) ◽  
pp. 3551-3558 ◽  
Author(s):  
Rebecca Munk Vejborg ◽  
Per Klemm
Keyword(s):  
Biofilm Formation ◽  
Fish Muscle ◽  
Physicochemical Characteristics ◽  
Economic Problem ◽  
Bacterial Attachment ◽  
Bacterial Biofilm ◽  
Adhesive Properties ◽  
Muscle Extract ◽  
Wide Range ◽  
Inert Surfaces

ABSTRACT Bacterial biofilm formation on inert surfaces is a significant health and economic problem in a wide range of environmental, industrial, and medical areas. Bacterial adhesion is generally a prerequisite for this colonization process and, thus, represents an attractive target for the development of biofilm-preventive measures. We have previously found that the preconditioning of several different inert materials with an aqueous fish muscle extract, composed primarily of fish muscle α-tropomyosin, significantly discourages bacterial attachment and adhesion to these surfaces. Here, this proteinaceous coating is characterized with regards to its biofilm-reducing properties by using a range of urinary tract infectious isolates with various pathogenic and adhesive properties. The antiadhesive coating significantly reduced or delayed biofilm formation by all these isolates under every condition examined. The biofilm-reducing activity did, however, vary depending on the substratum physicochemical characteristics and the environmental conditions studied. These data illustrate the importance of protein conditioning layers with respect to bacterial biofilm formation and suggest that antiadhesive proteins may offer an attractive measure for reducing or delaying biofilm-associated infections.

scholarly journals Effect of S-PRG Eluate on Biofilm Formation and Enzyme Activity of Oral Bacteria

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Masahiro Yoneda ◽  
Nao Suzuki ◽  
Yosuke Masuo ◽  
Akie Fujimoto ◽  
Kosaku Iha ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Composite Resin ◽  
Fusobacterium Nucleatum ◽  
Microtiter Plate ◽  
Oral Bacteria ◽  
Gelatin Film ◽  
Glass Ionomer ◽  
Microtiter Plate Assay ◽  
Substrate Hydrolysis ◽  
Adherence Ability

Recently, the antibacterial activity of a composite resin containing prereacted glass ionomer (S-PRG) filler was revealed. We examined the effect of an S-PRG eluate on various biologic activities ofStreptococcus mutansandPorphyromonas gingivalis. Adherence ability ofS. mutanswas evaluated by microtiter plate assay; protease and gelatinase activities ofP. gingivaliswere examined by synthetic substrate hydrolysis and gelatin film spot assay, respectively. Coaggregation ofP. gingivaliswithFusobacterium nucleatumwas also examined. S-PRG eluate was found to suppress streptococcal adherence. S-PRG eluate inhibited the protease and gelatinase activities ofP. gingivalisand the coaggregation betweenP. gingivalisandF. nucleatum. These results indicate that S-PRG eluate suppresses streptococcal adherence and inhibits the protease and coaggregation activities ofP. gingivalis. These findings may prompt research into novel strategies for preventing caries and periodontitis.

Relationship between halitosis and periodontal disease - associated oral bacteria in tongue coatings

2013 ◽  
Vol 12 (2) ◽  
pp. 145-151 ◽  
Author(s):  
T Amou ◽  
D Hinode ◽  
M Yoshioka ◽  
D Grenier
Keyword(s):  
Periodontal Disease ◽  
Oral Bacteria

scholarly journals The major outer sheath protein forms distinct conformers and multimeric complexes in the outer membrane and periplasm of Treponema denticola

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Robbins Puthenveetil ◽  
Sanjiv Kumar ◽  
Melissa J. Caimano ◽  
Abhishek Dey ◽  
Arvind Anand ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Treponema Denticola ◽  
Outer Sheath

scholarly journals Invasion of Oral and Aortic Tissues by Oral Spirochete Treponema denticola in ApoE−/−Mice Causally Links Periodontal Disease and Atherosclerosis

2014 ◽  
Vol 82 (5) ◽  
pp. 1959-1967 ◽  
Author(s):  
Sasanka S. Chukkapalli ◽  
Mercedes F. Rivera ◽  
Irina M. Velsko ◽  
Ju-Youn Lee ◽  
Hao Chen ◽  
...  
Keyword(s):  
Periodontal Disease ◽  
Alveolar Bone ◽  
Tissue Growth ◽  
Aortic Tissue ◽  
Treponema Denticola ◽  
Rapid Progression ◽  
Causative Role ◽  
Igg Antibody ◽  
Content Type ◽  
Periodontal Infection

ABSTRACTTreponema denticolais a predominantly subgingival oral spirochete closely associated with periodontal disease and has been detected in atherosclerosis. This study was designed to evaluate causative links between periodontal disease induced by chronic oralT. denticolainfection and atherosclerosis in hyperlipidemic ApoE−/−mice. ApoE−/−mice (n= 24) were orally infected withT. denticolaATCC 35404 and were euthanized after 12 and 24 weeks.T. denticolagenomic DNA was detected in oral plaque samples, indicating colonization of the oral cavity. Infection elicited significantly (P= 0.0172) higher IgG antibody levels and enhanced intrabony defects than sham infection.T. denticola-infected mice had higher levels of horizontal alveolar bone resorption than sham-infected mice and an associated significant increase in aortic plaque area (P≤ 0.05). Increased atherosclerotic plaque correlated with reduced serum nitric oxide (NO) levels and increased serum-oxidized low-density lipoprotein (LDL) levels compared to those of sham-infected mice.T. denticolainfection altered the expression of genes known to be involved in atherosclerotic development, including the leukocyte/endothelial cell adhesion gene (Thbs4), the connective tissue growth factor gene (Ctgf), and the selectin-E gene (Sele). Fluorescentin situhybridization (FISH) revealedT. denticolaclusters in both gingival and aortic tissue of infected mice. This is the first study examining the potential causative role of chronicT. denticolaperiodontal infection and vascular atherosclerosisin vivoin hyperlipidemic ApoE−/−mice.T. denticolais closely associated with periodontal disease and the rapid progression of atheroma in ApoE−/−mice. These studies confirm a causal link for active oralT. denticolainfection with both atheroma and periodontal disease.

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