Streptococcus mutans biofilm formation is dependent on extracellular DNA in primary low pH conditions

Kawarai, Taketo; Narisawa, Naoki; Suzuki, Yusuke; Nagasawa, Ryo; Senpuku, Hidenobu

doi:10.1016/j.job.2015.12.004

Cited by 25 publications

(21 citation statements)

References 44 publications

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“…Isolation and quantification of eDNA were performed as described elsewhere (46). Five hundred microliters of overnight culture was mixed with 4.5 ml of TSBs, TSBg, or TSBr in a 6-well culture plate (Iwaki, Tokyo, Japan).…”

Section: Methodsmentioning

confidence: 99%

“…After the biofilm formed in the 6-well plate, the culture supernatant was collected for quantification of soluble polysaccharides, and the biofilm attached to the bottom of the well was collected with a cell scraper for quantification of insoluble polysaccharides. We quantified the soluble and insoluble polysaccharides by the phenol-sulfuric acid method, as described elsewhere (46).…”

Section: Methodsmentioning

confidence: 99%

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Raffinose Induces Biofilm Formation by Streptococcus mutans in Low Concentrations of Sucrose by Increasing Production of Extracellular DNA and Fructan

Nagasawa

Sato

Senpuku

2017

Appl Environ Microbiol

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Streptococcus mutans is the primary etiological agent of dental caries and causes tooth decay by forming a firmly attached biofilm on tooth surfaces. Biofilm formation is induced by the presence of sucrose, which is a substrate for the synthesis of extracellular polysaccharides but not in the presence of oligosaccharides. Nonetheless, in this study, we found that raffinose, which is an oligosaccharide with an intestinal regulatory function and antiallergic effect, induced biofilm formation by S. mutans in a mixed culture with sucrose, which was at concentrations less than those required to induce biofilm formation directly. We analyzed the possible mechanism behind the small requirement for sucrose for biofilm formation in the presence of raffinose. Our results suggested that sucrose contributed to an increase in bacterial cell surface hydrophobicity and biofilm formation. Next, we examined how the effects of raffinose interacted with the effects of sucrose for biofilm formation. We showed that the presence of raffinose induced fructan synthesis by fructosyltransferase and aggregated extracellular DNA (eDNA, which is probably genomic DNA released from dead cells) into the biofilm. eDNA seemed to be important for biofilm formation, because the degradation of DNA by DNase I resulted in a significant reduction in biofilm formation. When assessing the role of fructan in biofilm formation, we found that fructan enhanced eDNA-dependent cell aggregation. Therefore, our results show that raffinose and sucrose have cooperative effects and that this induction of biofilm formation depends on supportive elements that mainly consist of eDNA and fructan.IMPORTANCE The sucrose-dependent mechanism of biofilm formation in Streptococcus mutans has been studied extensively. Nonetheless, the effects of carbohydrates other than sucrose are inadequately understood. Our findings concerning raffinose advance the understanding of the mechanism underlying the joint effects of sucrose and other carbohydrates on biofilm formation. Since raffinose has been reported to have positive effects on enterobacterial flora, research on the effects of raffinose on the oral flora are required prior to its use as a beneficial sugar for human health. Here, we showed that raffinose induced biofilm formation by S. mutans in low concentrations of sucrose. The induction of biofilm formation generally generates negative effects on the oral flora. Therefore, we believe that this finding will aid in the development of more effective oral care techniques to maintain oral flora health.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Raffinose Induces Biofilm Formation by Streptococcus mutans in Low Concentrations of Sucrose by Increasing Production of Extracellular DNA and Fructan

Nagasawa

Sato

Senpuku

2017

Appl Environ Microbiol

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“…1) (Allesen-Holm et al 2006; Böckelmann et al 2006; Flemming et al 2007). As a consequence, exDNA has been described as a structural component of the extracellular matrix, being essential especially during biofilm formation (Conover et al 2011; Kawarai et al 2016; Martins et al 2010; Novotny et al 2013; Nur et al 2013; Seper et al 2011; Whitchurch et al 2002; Zhao et al 2013) (reviewed by Flemming et al 2016; Montanaro et al 2011) and thus being actively secreted by the biofilm-producing microorganisms (Barnes et al 2012; Kilic et al 2017; Liao et al 2014; Rose and Bermudez 2016; Zafra et al 2012). A genome-wide screening for genes involved in exDNA release during biofilm formation by S. aureus was recently done (DeFrancesco et al 2017).…”

Section: Biofilmsmentioning

confidence: 99%

Extracellular DNA in natural environments: features, relevance and applications

Nagler

Insam

Pietramellara

et al. 2018

Appl Microbiol Biotechnol

180

151

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Extracellular DNA (exDNA) is abundant in many habitats, including soil, sediments, oceans and freshwater as well as the intercellular milieu of metazoa. For a long time, its origin has been assumed to be mainly lysed cells. Nowadays, research is collecting evidence that exDNA is often secreted actively and is used to perform a number of tasks, thereby offering an attractive target or tool for biotechnological, medical, environmental and general microbiological applications. The present review gives an overview on the main research areas dealing with exDNA, depicts its inherent origins and functions and deduces the potential of existing and emerging exDNA-based applications. Furthermore, it provides an overview on existing extraction methods and indicates common pitfalls that should be avoided whilst working with exDNA.

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“…On the other hand, DNase based treatments of pre-formed biofilms have not been explored much. There are only a few reports available [ 20 , 21 ], which describe application of DNase in combination with proteinase K [ 22 , 23 ], EDTA [ 24 ] and dextranase [ 25 ] on already formed biofilms. Such a biofilm control/therapeutic effect has been discussed using the term “Post-treatment” in this study.…”

Section: Introductionmentioning

confidence: 99%

Antibiofilm Effect of DNase against Single and Mixed Species Biofilm

Sharma¹,

Singh²

2018

Foods

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Biofilms are aggregates of microorganisms that coexist in socially coordinated micro-niche in a self-produced polymeric matrix on pre-conditioned surfaces. The biofilm matrix reduces the efficacy of antibiofilm strategies. DNase degrades the extracellular DNA (e-DNA) present in the matrix, rendering the matrix weak and susceptible to antimicrobials. In the current study, the effect of DNase I was evaluated during biofilm formation (pre-treatment), on preformed biofilms (post-treatment) and both (dual treatment). The DNase I pre-treatment was optimized for P. aeruginosa PAO1 (model biofilm organism) at 10 µg/mL and post-treatment at 10 µg/mL with 15 min of contact duration. Inclusion of Mg2+ alongside DNase I post-treatment resulted in 90% reduction in biofilm within only 5 min of contact time (irrespective of age of biofilm). On extension of these findings, DNase I was found to be less effective against mixed species biofilm than individual biofilms. DNase I can be used as potent antibiofilm agent and with further optimization can be effectively used for biofilm prevention and reduction in situ.

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Streptococcus mutans biofilm formation is dependent on extracellular DNA in primary low pH conditions

Cited by 25 publications

References 44 publications

Raffinose Induces Biofilm Formation by Streptococcus mutans in Low Concentrations of Sucrose by Increasing Production of Extracellular DNA and Fructan

Raffinose Induces Biofilm Formation by Streptococcus mutans in Low Concentrations of Sucrose by Increasing Production of Extracellular DNA and Fructan

Extracellular DNA in natural environments: features, relevance and applications

Antibiofilm Effect of DNase against Single and Mixed Species Biofilm

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