Effect of Commonly Prescribed Liquid Medications on Streptococcus mutans Biofilm. An in vitro study

Objective: This study addressed the effect of pediatric liquid antibiotic medications on Streptococcus mutans UA159. These suspensions commonly contain sugars such as sucrose to make them more palatable for children. The study was designed to evaluate the effects of oral liquid antibiotics on Streptococcus mutans growth and biofilm formation. Study Design: A 24 hour culture of S. mutans was treated with various concentrations of liquid medications commonly prescribed to children for odontogenic or fungal infections– amoxicillin, penicillin VK, clindamycin, and nystatin. The study was conducted in sterile 96-well flat bottom microtiter plates. The minimum inhibitory and biofilm inhibitory concentrations (MIC/MBIC) of S. mutans were determined for each medication. S. mutans was cultured with and without the test drugs, the amount of total growth measured, the biofilms washed, fixed, and stained with crystal violet. The absorbance was determined to evaluate biofilm formation. Results: Higher concentrations of amoxicillin, penicillin VK and clindamycin had decreased biofilm and overall growth than the control. The MICs were 1:2,560 (1.95 ug/ml), 1:2,560 (1.95 ug/ml) and 1:40 (9.375 ug/ml), while the MBIC were 1:640 (7.8 ug/ml), 1:1,280 (3.9 ug/ml) and 1:20 (18.75 ug/ml), respectively. Lower concentrations provided increased biofilm and overall growth. Nystatin induced significantly more biofilm and overall growth than the control at all concentrations. Conclusion: At high concentrations, approximately at the levels expected to be present in the oral cavity of children, amoxicillin, penicillin, and clindamycin inhibited S. mutans biofilm and overall growth due to their antibiotic activity, while at lower concentrations the three antibiotics demonstrated an increase in biofilm and growth. The increase in S. mutans biofilm and overall growth is most likely attributed to the sugar content in the medications. Nystatin provided an increase in biofilm and growth at each concentration tested.

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Copper Supplementation in Watering Solution Reaches the Xylem But Does Not Protect Tobacco Plants Against Xylella fastidiosa Infection

Plant Disease ◽

10.1094/pdis-08-19-1748-re ◽

2020 ◽

Vol 104 (3) ◽

pp. 724-730 ◽

Cited By ~ 2

Author(s):

Qing Ge ◽

Paul A. Cobine ◽

Leonardo De La Fuente

Keyword(s):

Biofilm Formation ◽

Inductively Coupled Plasma ◽

Tap Water ◽

Xylella Fastidiosa ◽

In Vitro Study ◽

Optical Emission ◽

In Planta ◽

Inductively Coupled ◽

High Concentrations

Xylella fastidiosa is a xylem-limited plant pathogenic bacterium that causes disease in many crops worldwide. Copper (Cu) is an antimicrobial agent widely used on X. fastidiosa hosts to control other diseases. Although the effects of Cu for control of foliar pathogens are well known, it is less studied on xylem-colonizing pathogens. Previous results from our group showed that low concentrations of CuSO4 increased biofilm formation, whereas high concentrations inhibited biofilm formation and growth in vitro. In this study, we conducted in planta experiments to determine the influence of Cu in X. fastidiosa infection using tobacco as a model. X. fastidiosa-infected and noninfected plants were watered with tap water or with water supplemented with 4 mM or 8 mM of CuSO4. Symptom progression was assessed, and sap and leaf ionome analysis was performed by inductively coupled plasma with optical emission spectroscopy. Cu uptake was confirmed by increased concentrations of Cu in the sap of plants treated with CuSO4-amended water. Leaf scorch symptoms in Cu-supplemented plants showed a trend toward more severe at later time points. Quantification of total and viable X. fastidiosa in planta indicated that CuSO4-amended treatments did not inhibit but slightly increased the growth of X. fastidiosa. Cu in sap was in the range of concentrations that promote X. fastidiosa biofilm formation according to our previous in vitro study. Based on these results, we proposed that the plant Cu homeostasis machinery controls the level of Cu in the xylem, preventing it from becoming elevated to a level that would lead to bacterial inhibition.

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Effect of Human Milk and its Components on Streptococcus Mutans Biofilm Formation

Journal of Clinical Pediatric Dentistry ◽

10.17796/1053-4628-39.3.255 ◽

2015 ◽

Vol 39 (3) ◽

pp. 255-261 ◽

Cited By ~ 11

Author(s):

LM Allison ◽

LA Walker ◽

BJ Sanders ◽

Z Yang ◽

G Eckert ◽

...

Keyword(s):

Breast Milk ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

Current Knowledge ◽

Post Partum ◽

Human Breast Milk ◽

Human Breast ◽

Flat Bottom ◽

Biofilm Growth ◽

Microtiter Plates

Objective: This study investigated the effects of human breast milk and its components on the nutritional aspect of the caries process due to Streptococcus mutans UA159 biofilm formation. Study design: Human breast milk was collected from 11 mothers during 3-9 months postpartum. To test for the effect on biofilm formation, a 16-hour culture of S. mutans was treated with dilutions of human breast milk and several major components of human breast milk, lactose, lactoferrin, IgA, and bovine casein in sterile 96-well flat bottom microtiter plates for 24 hours. The biofilms were fixed, washed, stained with crystal violet, and extracted. Absorbance was measured to evaluate biofilm growth mass. Results: Dilutions 1:10-1:2,560 of the human breast milk samples increased biofilm formation by 1.5-3.8 fold compared to the control. Lactoferrin decreased biofilm formation significantly in all dilutions (average milk concentration of 3 mg/ml). Lactose had no effect at average breast milk concentrations (60 mg/ml) except at its lowest concentration (15 mg/ml) where it was increased. IgA significantly decreased biofilm formation at its highest concentration of 2,400 μg/ml (average milk concentration 600 μg/ml). Casein caused significantly increased biofilm formation at all concentrations tested above the average milk content (2.3 mg/ml). Conclusions: The results of this study demonstrate an increase in S. mutans biofilm formation by human breast milk 3-9 months post partum. Among its major components, only casein significantly increased biofilm formation among the concentrations analyzed. Lactose had no effect except at 15 mg/ml. Lactoferrin and IgA significantly decreased S. mutans biofilm formation at their highest concentrations. This information expands the current knowledge regarding the nutritional influence of breastfeeding and validates the necessity to begin an oral hygiene regimen once the first tooth erupts.

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Antibiofilm activity and cytotoxicity of silk sericin against Streptococcus mutans bacteria in biofilm: an in vitro study

Journal of Wound Care ◽

10.12968/jowc.2020.29.sup4.s25 ◽

2020 ◽

Vol 29 (Sup4) ◽

pp. S25-S35

Author(s):

Pornanong Aramwit ◽

Supamas Napavichayanum ◽

Prompong Pienpinijtham ◽

Yousef Rasmi ◽

Nipaporn Bang

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

In Vitro Study ◽

Bacterial Biofilm ◽

Vitro Study ◽

Antibiofilm Activity ◽

Diphenyltetrazolium Bromide ◽

Infrared Spectrometer ◽

Disruption Effect

Objective: To investigate the potential of sericin extracted by different methods to inhibit biofilm formation (prevention) and disrupt already formed biofilm (treatment). Method: In this in vitro study, sericin was extracted by heat, acid, alkali and urea. Streptococcus mutans bacteria were cultivated in the presence of various concentrations of sericin to evaluate antibiofilm formation using cell density assay (inhibition effect before biofilm formed). Conversely, various concentrations of sericin were added to a biofilm already formed by Streptococcus mutans bacteria, and the viability of bacteria assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (disruption effects after biofilm formed). Structures of extracted sericin were evaluated using circular dichroism and Fourier-transform infrared spectrometer. Results: The urea-extracted sericin at all concentrations (12.5mg/ml, 25mg/ml, 50mg/ml and 100mg/ml) showed the highest potential antibiofilm activity in terms of both inhibition and disruption effects, compared with sericin extracted by heat, acid or alkali. The heat-extracted and acid-extracted sericin were found to reduce the biofilm formation dose-dependently, while the alkali-extracted sericin did not show either inhibition or disruption effect on the bacterial biofilm. The urea-extracted sericin also killed the bacteria residing within the biofilm, possibly due to its modified structure which may destabilise the bacterial cell wall, leading to membrane disintegration and, finally, cell death. Conclusion: Our results demostrated the antibiofilm activity of sericin. This could form the basis of further research on the mechanism and application of sericin as a novel antibiofilm agent.

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BIOFILM FORMATION OF CANDIDA ALBICANS EXPOSED TO ETHANOL EXTRACT OF PROPOLIS

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2019.v11s1.18344 ◽

2019 ◽

pp. 210-213

Author(s):

ARIADNA ADISATTYA DJAIS ◽

JEMMY . ◽

NADHIFA PUTRI ◽

ANDIN RAHMANIA PUTRI ◽

RISQA RINA DARWITA ◽

...

Keyword(s):

Candida Albicans ◽

Biofilm Formation ◽

Fungal Infections ◽

Ethanol Extract ◽

Clinical Strain ◽

Propolis Extract ◽

Microtiter Plates ◽

Treatment And Prevention ◽

Student’S T

Objective: Propolis extract showed an excellent in vitro performance against yeast and was additionally found to be fungistatic and fungicidal. Propolisextract is also used for treatment and prevention of fungal infections. However, its effectiveness against Candida albicans biofilm formation requiresinvestigation. The study evaluated the ability of propolis to inhibit C. albicans while the fungus is growing as a biofilm in vitro.Methods: Two reference strains, C. albicans ATCC 25923 and a clinical strain (laboratory stock), were used in this study. For the biofilm experiment,the fungi were cultured in Tryptic Soy Broth medium with 1% sucrose and incubated at 37°C for 24 h, and different concentrations of ethanol extractof propolis were used as the inhibitor agents. Biofilm assays were performed in 96-well microtiter plates, quantification of the total biofilm biomasswas performed using a crystal violet staining method, and the Student’s t-test was chosen for statistical analyses.Results: Our data showed that 3 h incubation with propolis did not affect the biomass in the experimental group compared to the control. When theincubation time was extended to 18 h, the biomass increased significantly compared to the control.Conclusion: This study showed that several concentrations of propolis did not inhibit biofilm. However, in each incubation time, we observed nohyphal morphology in the biofilm mass. Propolis might attenuate the opportunistic virulence of fungus growing as a biofilm in vitro. Further studiesare necessary to confirm this phenomenon.

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THE EFFECT OF CURCUMA XANTHORRHIZA - ETHANOL EXTRACT TO BIOFILM FORMATION OF STREPTOCOCCUS MUTANS AND AGGREGATIBACTER ACTINOMYCETEMCOMITANS (DENTAL BIOFILM RESEARCH: IN VITRO STUDY)

International Journal of Applied Pharmaceutics ◽

10.22159/ijap.2017.v9s1.14_16 ◽

2017 ◽

Vol 9 ◽

pp. 20

Author(s):

Fidhianissa . . ◽

Hedijanti Joenoes ◽

Ariadna A Djais

Keyword(s):

Streptococcus Mutans ◽

Biofilm Formation ◽

In Vitro Study ◽

Ethanol Extract ◽

Single Species ◽

Vitro Study ◽

Bacterial Suspension ◽

Dental Biofilm ◽

Curcuma Xanthorrhiza

Objective: This in vitro study aimed to analyze the mass ratio of single- and dual-species Streptococcus mutans and Aggregatibacteractinomycetemcomitans biofilm after exposure to Curcuma xanthorrhiza ethanol extract (Xan).Methods: A bacterial suspension in brain heart infusion medium, enriched with 0.2% sucrose, was exposed to the Xan, incubated for 18 hrs, andanalyzed using a crystal violet assay.Results: This research concluded that the minimum inhibitory concentration of ethanol-temulawak extract against S. mutans was 5%, while theminimum bactericidal concentration was 15%.Conclusions: Xan prevented biofilm formation of single-species S. mutans and dual-species S. mutans and A. actinomycetemcomitans more effectivelythan it did single-species A. actinomycetemcomitans.

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The effect of photodynamic therapy by gold nanoparticles on Streptococcus mutans and biofilm formation: an in vitro study

Lasers in Medical Science ◽

10.1007/s10103-021-03422-x ◽

2021 ◽

Author(s):

Fatemeh Lavaee ◽

Mohammad Motamedifar ◽

Ghazal Rafiee

Keyword(s):

Gold Nanoparticles ◽

Photodynamic Therapy ◽

Streptococcus Mutans ◽

Biofilm Formation ◽

In Vitro Study ◽

Vitro Study

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Voice Prosthesis Coated with Sustained Release Varnish Containing Clotrimazole Shows Long-Term Protection against Candida albicans: An In Vitro Study

Molecules ◽

10.3390/molecules26175395 ◽

2021 ◽

Vol 26 (17) ◽

pp. 5395

Author(s):

Ronit Vogt Sionov ◽

Irith Gati ◽

David Kirmayer ◽

Michael Friedman ◽

Doron Steinberg ◽

...

Keyword(s):

Candida Albicans ◽

Sustained Release ◽

Biofilm Formation ◽

Fungal Infections ◽

In Vitro Study ◽

Inhibition Zone ◽

Voice Prosthesis ◽

Major Health Problem

Fungal biofilm formation on voice prosthesis (VP) is a major health problem that requires repeated replacement of the prosthesis. Candida albicans is one of the pathogens that frequently inhabits the VP. We proposed that coating VPs with sustained-release varnish (SRV) containing clotrimazole (CTZ) might prevent fungal biofilm formation. The long-term antifungal activities of SRV-CTZ- versus SRV-placebo-coated VPs was tested daily by measuring the inhibition zone of C. albicans seeded on agar plates or by measuring the fungal viability of C. albicans in suspension. The extent of biofilm formation on coated VPs was analyzed by confocal microscopy and scanning electron microscopy. We observed that SRV-CTZ-coated VPs formed a significant bacterial inhibition zone around the VPs and prevented the growth of C. albicans in suspension during the entire testing period of 60 days. Fungal biofilms were formed on placebo-coated VPs, while no significant biofilms were observed on SRV-CTZ-coated VPs. HPLC analysis shows that CTZ is continuously released during the whole test period of 60 days at a concentration above the minimal fungistatic concentration. In conclusion, coating VPs with an SRV-CTZ film is a potential effective method for prevention of fungal infections and biofilm formation on VPs.

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Effect of Honey on Streptococcus mutans Growth and Biofilm Formation

Applied and Environmental Microbiology ◽

10.1128/aem.05538-11 ◽

2011 ◽

Vol 78 (2) ◽

pp. 536-540 ◽

Cited By ~ 36

Author(s):

Hani M. Nassar ◽

Mingyun Li ◽

Richard L. Gregory

Keyword(s):

Streptococcus Mutans ◽

Bacterial Growth ◽

Biofilm Formation ◽

Sugar Content ◽

Antibacterial Properties ◽

Maximum Velocity ◽

Content Type ◽

Microtiter Plates ◽

High Sugar Content ◽

Natural Honey

ABSTRACTBecause of the tradition of using honey as an antimicrobial medicament, we investigated the effect of natural honey (NH) onStreptococcus mutansgrowth, viability, and biofilm formation compared to that of an artificial honey (AH). AH contained the sugars at the concentrations reported for NH. NH and AH concentrations were obtained by serial dilution with tryptic soy broth (TSB). Several concentrations of NH and AH were tested for inhibition of bacterial growth, viability, and biofilm formation after inoculation withS. mutansUA159 in 96-well microtiter plates to obtain absorbance and CFU values. Overall, NH supported significantly less (P< 0.05) bacterial growth than AH at 25 and 12.5% concentrations. At 50 and 25% concentrations, both honey groups provided significantly less bacterial growth and biofilm formation than the TSB control. For bacterial viability, the results for all honey concentrations except 50% NH were not significantly different from those for the TSB control. NH was able to decrease the maximum velocity ofS. mutansgrowth compared to AH. In summary, NH demonstrated more inhibition of bacterial growth, viability, and biofilm formation than AH. This study highlights the potential antibacterial properties of NH and could suggest that the antimicrobial mechanism of NH is not solely due to its high sugar content.

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