Real-time monitoring of the metabolic activity of periodontopathic bacteria

Ishiguro, Kazuko; Washio, Jumpei; Sasaki, Keiichi; Takahashi, Nobuhiro

doi:10.1016/j.mimet.2015.05.015

Cited by 17 publications

(17 citation statements)

References 25 publications

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“…After that, biofilms of strains were carefully washed twice with PBS, and biofilm biomass was determined using the crystal violet (CV) assay described by Sabaeifard [11]. Viability of biofilms formed by strains was evaluated by the Alamar Blue® assay [12]. The method is based on the dye resazurin.…”

Section: Methodsmentioning

confidence: 99%

Genetic polymorphism of scrA gene of Streptococcus mutans isolates is not associated with biofilm formation in severe early childhood caries

Zhou

Tao

et al. 2017

BMC Oral Health

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BackgroundTo explore and analyse the association between biofilm and the genetic polymorphisms of scrA gene of EnzymeIIscr found in clinical isolates of Streptococcus mutans (S. mutans) from severe early childhood caries (S-ECC) in 3 years old children.MethodsClinical strains of S. mutans were conserved from a previous study. Thirty strains of S. mutans from the S-ECC group and 30 strains of S. mutans from the caries free (CF) group were selected. Biomass and viability of biofilm formed by the strains were evaluated by crystal violet and alamar blue assay. Genomic DNA was extracted from the S. mutans isolates. PCR was conducted to amplify scrA gene. After purified and sequenced the PCR products, BioEdit sofeware was used to analyse the sequence results. A chi-square test was used to compare the results.ResultsCompared to the CF group, the biomass of S-ECC group was higher (P = 0.0424). However, the viability of the two groups showed no significant difference. All 60 clinically isolated S. mutans strains had a 1995 base pair (bp) scrA gene. Forty-nine point mutations were identified in scrA from the 60 clinical isolates. There were 17 missense point mutations at the 10, 65, 103, 284, 289, 925, 1444, 1487, 1494, 1508, 1553, 1576, 1786, 1822, 1863, 1886, and 1925 bp positions. The other 32 mutations were silent point mutations. No positions were found at active sites of ScrA. The statistic analyse showed no significant missense mutation rates between the two groups.ConclusionsThere was no association between biofilm and genetic polymorphisms of scrA from S. mutans with S-ECC in 3 years old children.Electronic supplementary materialThe online version of this article (doi:10.1186/s12903-017-0407-0) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Genetic polymorphism of scrA gene of Streptococcus mutans isolates is not associated with biofilm formation in severe early childhood caries

Zhou

Tao

et al. 2017

BMC Oral Health

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“…Resazurin has been used for testing milk (Reinheimer and Demkow ) as an early indicator of the growth of mycobacteria (Bastian et al ) and in assessing the activities of antibacterial compounds (Sarker et al ; Gavrish et al ). Research has demonstrated the use of resazurin for real‐time monitoring of the metabolic activity of oral bacteria (Wei et al ; Tawakoli et al ; Suntar et al ), with a recent study validating its use for periodontopathic and cariogenic bacteria in vitro (Ishiguro et al ).…”

Section: Discussionmentioning

confidence: 99%

A rapid chair‐side method for the estimation of oral bacterial colonization density

et al. 2019

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Aims Caries and periodontal disease are associated with inadequate control of oral bacteria. Since conventional microbiological evaluations are impractical in dental clinics or public engagement activities, a rapid test for the quantification of oral bacteria represents a useful tool. We describe the development of a colour change test to rapidly estimate bacterial colonisation density in the mouth. Methods and Results Volunteers rinsed with milk or milkshake. Viability indicators were added and colour changes quantified during incubation. Using milkshake and the resazurin‐based solution PrestoBlue (9% v/v), the method distinguished between samples before and after brushing within 5 min. Colour changes were quantified and viable counts were obtained using oral rinses. Measured colour changes strongly correlated with total counts of both anaerobes and streptococci (Spearman’s correlation coefficient of 0·782 and 0·769, respectively, P ≤ 0·001) and with perceived changes, as determined by volunteers (n = 10) visually ranking images. Conclusions The resazurin milkshake test can rapidly and visually quantify viable bacteria in oral samples. Significance and Impact of the Study The resazurin milkshake test could serve as a sensitive semi‐quantitative method for measuring oral bacteria in human oral rinse samples.

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“…There was some impairment in the chemical reactions responsible for metabolizing carbohydrate sources after violet-blue light treatment contributing to the inhibition of metabolic activities in S. mutans biofilm cells seen in this study. The pathogenic nature of cariogenic biofilm is determined by metabolic activity (21). Dehydrogenases in bacterial cells play a vital role in metabolic activities.…”

Section: Discussionmentioning

confidence: 99%

Effect of phototherapy on the metabolism of <i>Streptococcus mutans</i> biofilm based on a colorimetric tetrazolium assay

Gomez

Huang

Eckert

et al. 2018

Journal of Oral Science

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The aim of this in vitro study was to determine the effect of violet-blue light on the metabolic activity of early Streptococcus mutans biofilm, reincubated at 0, 2, and 6 h after 5 min of violet-blue light treatment. S. mutans UA159 biofilm cells were cultured for 12 to 16 h in microtiter plates with Tryptic Soy broth (TSB) or TSB with 1% sucrose (TSBS) and irradiated with violet-blue light for 5 min. After irradiation, the plates were reincubated at 37°C for 0, 2, or 6 h in 5% CO 2 . Colorimetric tetrazolium salt reduction assay was used to investigate bacterial metabolic activity. Mixed model ANOVA was used to find the difference between the violet-blue light treated and nontreated groups. Bacterial metabolic activity was significantly lower in the violet-blue light group for TSB than in the nontreated group (P < 0.0001) regardless of recovery time. However, the differences between metabolic activity in the treated groups without sucrose decreased over time. For TSBS, metabolic activity was significantly lower with violet-blue light at 0 and 2 h. Violet-blue light inhibited the metabolic activity of S. mutans biofilm cells in the light-treated group. This finding may present a unique treatment method for patients with active caries.

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Real-time monitoring of the metabolic activity of periodontopathic bacteria

Cited by 17 publications

References 25 publications

Genetic polymorphism of scrA gene of Streptococcus mutans isolates is not associated with biofilm formation in severe early childhood caries

Genetic polymorphism of scrA gene of Streptococcus mutans isolates is not associated with biofilm formation in severe early childhood caries

A rapid chair‐side method for the estimation of oral bacterial colonization density

Effect of phototherapy on the metabolism of <i>Streptococcus mutans</i> biofilm based on a colorimetric tetrazolium assay

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