scholarly journals In vitro Antibiofilm Activity of Pomegranate (Punica granatum) Juice on Oral Pathogens

2019 ◽  
Vol 1 (2) ◽  
pp. 49
Author(s):  
Jemima Pramadita ◽  
Armelia Sari Widyarman
Keyword(s):  
Biofilm Formation ◽  
Brain Heart Infusion ◽  
Punica Granatum ◽  
Pomegranate Juice ◽  
Bacterial Suspension ◽  
Antibiofilm Activity ◽  
Microplate Reader ◽  
Level Of Significance ◽  
Negative Controls

Introduction: Pomegranate (Punica granatum) fruit contains valuable ingredients, such as ellagitannins and flavonoids, that have many potential effects, including antibacterial, antifungal, and anti-inflammatory functions. Objectives: The aim of this study was to investigate the effects of pomegranate fruit juice on F. nucleatum and S. sanguinis monospecies and multispecies biofilm formation in vitro. Methods: Pomegranate juice was obtained using a juicer and diluted using a brain heart infusion (BHI) broth into five different concentrations. The biofilm assay was performed as follows: F. nucleatum and S. sanguinis were cultured separately in the BHI broth for 48 hours at 37°C in an anaerobic atmosphere. A 200 mL bacterial suspension (107 CFU/mL) was distributed into a 96-well plate and incubated for 24 hours to form  a biofilm. Subsequently, pomegranate juice was added to the biofilm well and observed after 1 hours, 3 hours, 6 hours, and 24 hours. The biofilm mass was measured using a microplate reader (490 nm) after crystal violet staining. Chlorhexidine (0.2%) and the biofilms without treatment were used as the positive and negative controls, respectively. The data were statistically analyzed using one-way analysis of variance, with p<0.05 as the level of significance. Result: There was a significant biofilm reduction after treatment with pomegranate juice for all the concentrations and incubation times (p<0.05). The effective concentrations to inhibit the biofilm monospecies F. nucleatum and S. sanguinis and the multispecies were 6.25% (OD 0.148±0.019), 50% (OD 0.211±0.026), and 6.25% (OD 0.024±0.209), respectively. Conclusion: Pomegranate juice inhibits F. nucleatum and S. sanguinis biofilm formation as a monospecies and a multispecies. Future studies are needed to observe the mechanism of this active substance.

scholarly journals In vitro Antibiofilm Activity of Pomegranate (Punica granatum) Juice on Oral Pathogens

2019 ◽  
Vol 2 (1) ◽  
pp. 15
Author(s):  
Jemima Pramadita ◽  
Armelia Sari Widyarman
Keyword(s):  
Biofilm Formation ◽  
Brain Heart Infusion ◽  
Punica Granatum ◽  
Pomegranate Juice ◽  
Bacterial Suspension ◽  
Antibiofilm Activity ◽  
Microplate Reader ◽  
Level Of Significance ◽  
Negative Controls

  Introduction: Pomegranate (Punica granatum) fruit contains valuable ingredients, such as ellagitannins and flavonoids, that have many potential effects, including antibacterial, antifungal, and anti-inflammatory functions.   Objectives: The aim of this study was to investigate the effects of pomegranate fruit juice on F. nucleatum and S. sanguinis monospecies and multispecies biofilm formation in vitro.   Methods: Pomegranate juice was obtained using a juicer and diluted using a brain heart infusion (BHI) broth into five different concentrations. The biofilm assay was performed as follows: F. nucleatum and S. sanguinis were cultured separately in the BHI broth for 48 hours at 37°C in an anaerobic atmosphere. A 200 mL bacterial suspension (107 CFU/mL) was distributed into a 96-well plate and incubated for 24 hours to form  a biofilm. Subsequently, pomegranate juice was added to the biofilm well and observed after 1 hours, 3 hours, 6 hours, and 24 hours. The biofilm mass was measured using a microplate reader (490 nm) after crystal violet staining. Chlorhexidine (0.2%) and the biofilms without treatment were used as the positive and negative controls, respectively. The data were statistically analyzed using one-way analysis of variance, with p<0.05 as the level of significance.   Result: There was a significant biofilm reduction after treatment with pomegranate juice for all the concentrations and incubation times (p<0.05). The effective concentrations to inhibit the biofilm monospecies F. nucleatum and S. sanguinis and the multispecies were 6.25% (OD 0.148±0.019), 50% (OD 0.211±0.026), and 6.25% (OD 0.024±0.209), respectively.   Conclusion: Pomegranate juice inhibits F. nucleatum and S. sanguinis biofilm formation as a monospecies and a multispecies. Future studies are needed to observe the mechanism of this active substance.

scholarly journals The Anti-biofilm Activity of Nanometric Zinc doped Bioactive Glass against Putative Periodontal Pathogens: An in vitro Study

2018 ◽  
Vol 4 (1) ◽  
pp. 95-107
Author(s):  
Nasrin Esfahanizadeh ◽  
Mohammad Reza Nourani ◽  
Abbas Bahador ◽  
Nasrin Akhondi ◽  
Mostafa Montazeri
Keyword(s):  
Biofilm Formation ◽  
Sol Gel ◽  
Antimicrobial Properties ◽  
In Vitro Study ◽  
Inhibitory Effect ◽  
Antibiofilm Activity ◽  
Periodontal Pathogens ◽  
Microplate Reader ◽  
45S5 Bioglass

Abstract Colonization of periodontal pathogens on the surgical sites is one of the primary reasons for the failure of regenerative periodontal therapies. Bioactive glasses (BGs) owing to their favorable structural and antimicrobial properties have been proposed as promising materials for the reconstruction of periodontal and peri-implant bone defects. This study aimed to investigate the antibiofilm activity of zinc-doped BG (Zn/BG) compared with 45S5 Bioglass® (BG®) on putative periodontal pathogens. In this in vitro experimental study, the nano BG doped with 5-mol% zinc and BG® were synthesized by sol-gel method. Mono-species biofilms of Aggregatibacter actinomycetemcomitans (A. a), Porphyromonas gingivalis (P. g), and Prevotella intermedia (P. i)were prepared separately in a well-containing microplate. After 48 hours of exposure to generated materials at 37°C, the anti-biofilm potential of the samples was studied by measuring the optical density (OD) at 570nm wavelengths with a microplate reader. Two-way ANOVA then analyzed the results. Both Zn/BG and BG® significantly reduced the biofilm formation ability of all examined strains after 48 hours of incubation (P=0.0001). Moreover, the anti-biofilm activity of Zn/BG was significantly stronger than BG® (P=0.0001), which resulted in the formation of a weak biofilm (OD<1) compared with a moderately adhered biofilm observed with BG® (1<OD<2). Zn/BG showed a significant inhibitory effect on the biofilm formation of all examined periodontal pathogens. Given the enhanced regenerative and anti-biofilm properties of this novel biomaterial, further investigations are required for its implementation in clinical situations.

scholarly journals Potential of Lactobacillus casei shirota’s strain against the biofilm-forming of Salmonella Spp

2019 ◽  
Vol 8 (2) ◽  
pp. 54-63
Author(s):  
Annisa Rizka Fauziah ◽  
Meiska Bahar ◽  
Aprilla Ayu Wulandari
Keyword(s):  
Biofilm Formation ◽  
Lactobacillus Casei ◽  
Microtiter Plate ◽  
Assay Method ◽  
Antibiofilm Activity ◽  
Salmonella Spp ◽  
Microplate Reader ◽  
Competitive Adhesion ◽  
Concentration Series

Biofilm of Salmonella spp. is formed through the expression of biofilm genes associated with proteins (bapA) regulated by curli synthesis genes (csg) which carry out adhesion, colonization, maturation, and dispersion on the surface of the intestinal epithelium. This study aimed to determine the antibiofilm activity of Lactobacillus casei Shirota’S strain (LcS) as an inhibitor of Salmonella spp. biofilm formation in vitro. This research was a true experimental study using Microtiter Plate 96 wells Biofilm Assay method. The sample used was the suspension of Salmonella spp. The treatment was in the form of adding a LcS suspension with a concentration series of 10-1;10-2; 10-3;10-4; and 10-5. Biofilm measurements were carried out using a microplate reader and obtained quantitative data in the form of Optical Density at a wavelength of 595nm. The results of this study showed that LcS suspension has antibiofilm activity ranging from 10-5 concentrations with a percentage of 36.58% (p<0.05). The results of exometabolism LcS can reduce Salmonella growth. Exopolysaccharide (EPS) and sortase-dependent proteins (SrtA) of LcS form barriers as competitive adhesion in inhibiting pathogenic biofilm formation.

scholarly journals In Vitro Inhibition of Biofilm Formation by Staphylococcus Aureus Under the Action of Selected Plant Extracts

2019 ◽  
Vol 63 (1) ◽  
pp. 48-53 ◽  
Author(s):  
A. Piegerová ◽  
J. Koščová ◽  
P. Schusterová ◽  
R. Nemcová ◽  
M. Kryvtsova
Keyword(s):  
Staphylococcus Aureus ◽  
Plant Extracts ◽  
Biofilm Formation ◽  
Culture Media ◽  
Rosemary Extract ◽  
Bacterial Suspension ◽  
Antibiofilm Activity ◽  
Tryptone Soya Broth ◽  
Crystal Violet Dye

Abstract In our study we investigated the ability of selected plant extracts to inhibit the formation of biofilms produced by Staphylococcus aureus. In the first phase, we focused on the optimisation of conditions for the correct method of an approach. For optimisation, we standardized the culture media and the bacterial culture in order to obtain interpretable results. The TSB (Tryptone Soya Broth) medium was used for the preparation of an inoculum from the bacterial suspension. For the in vitro tests of antibiofilm activity against the species Staphylococcus aureus CCM 3953, we used propylene glycol (PG) plant extracts from sage and rosemary, prepared in three different concentrations of 0.01 %, 0.05 % and 0.1 %. The tests were implemented in microtitre plates using crystal violet dye at 0.1 % concentration for visualization of the intensity of a biofilm. The results were obtained, by spectrophotometric measurements at a wavelength of 550 nm. Both rosemary and sage plant extracts had a significant effect on the formation of a biofilm by S. aureus. The antibiofilm activity was concentration-dependent as the formation of biofilm was reduced more effectively with increasing concentration of the extracts. The best antibiofilm activity was observed with 0.1 % rosemary extract resulting in 94 % inhibition of the biofilm formation.

scholarly journals Antimicrobial and Antibiofilm Activity against Staphylococcus aureus of Opuntia ficus-indica (L.) Mill. Cladode Polyphenolic Extracts

Antioxidants ◽  
2019 ◽  
Vol 8 (5) ◽  
pp. 117 ◽  
Author(s):  
Federica Blando ◽  
Rossella Russo ◽  
Carmine Negro ◽  
Luigi De Bellis ◽  
Stefania Frassinetti
Keyword(s):  
Staphylococcus Aureus ◽  
Antioxidant Activity ◽  
Antioxidant Capacity ◽  
Biofilm Formation ◽  
Ex Vivo ◽  
Trolox Equivalent Antioxidant Capacity ◽  
Total Phenolic ◽  
Antibiofilm Activity ◽  
Opuntia Ficus Indica

Plant extracts are a rich source of natural compounds with antimicrobial properties, which are able to prevent, at some extent, the growth of foodborne pathogens. The aim of this study was to investigate the potential of polyphenolic extracts from cladodes of Opuntia ficus-indica (L.) Mill. to inhibit the growth of some enterobacteria and the biofilm formation by Staphylococcus aureus. Opuntia ficus-indica cladodes at two stages of development were analysed for total phenolic content and antioxidant activity by Oxygen Radical Absorbance Capacity (ORAC) and Trolox equivalent antioxidant capacity (TEAC) (in vitro assays) and by cellular antioxidant activity in red blood cells (CAA-RBC) (ex vivo assay). The Liquid Chromatography Time-of-Flight Mass Spectrometry (LC/MS–TOF) analysis of the polyphenolic extracts revealed high levels of piscidic acid, eucomic acid, isorhamnetin derivatives and rutin, particularly in the immature cladode extracts. Opuntia cladodes extracts showed a remarkable antioxidant activity (in vitro and ex vivo), a selective inhibition of the growth of Gram-positive bacteria, and an inhibition of Staphylococcus aureus biofilm formation. Our results suggest and confirm that Opuntia ficus-indica cladode extracts could be employed as functional food, due to the high polyphenolic content and antioxidant capacity, and used as natural additive for food process control and food safety.

scholarly journals In VitroAntibacterial Activity of Pomegranate Juice and Peel Extracts on Cariogenic Bacteria

2017 ◽  
Vol 2017 ◽  
pp. 1-7 ◽  
Author(s):  
Gianmaria Fabrizio Ferrazzano ◽  
Elisa Scioscia ◽  
Daniela Sateriale ◽  
Gabiria Pastore ◽  
Roberta Colicchio ◽  
...  
Keyword(s):  
Dental Caries ◽  
Clinical Isolate ◽  
Inhibitory Activity ◽  
Punica Granatum ◽  
Pomegranate Juice ◽  
Tooth Decay ◽  
Growth And Survival ◽  
Cariogenic Bacteria ◽  
Etiologic Agents

Aim. To evaluate the antimicrobial activity of hydroalcoholic extracts of pomegranate (Punica granatumL.) peel and juice, against the microorganisms considered the main etiologic agents of dental caries.Methods. The values of the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined againstStreptococcus mutansClarke ATCC® 25175™ strain andRothia dentocariosaclinical isolate.Results. Peel extracts inhibit effectively the growth and survival ofS. mutansATCC 25175 strain andR. dentocariosaclinical isolate with MIC and MBC values of 10 μg/μl and 15 μg/μl, respectively. Furthermore, the pomegranate juice extract showed high inhibitory activity againstS. mutansATCC 25175 strain with a MIC value of 25 μg/μl and a MBC value of 40 μg/μl, whereas, againstR. dentocariosa, it has displayed a moderate inhibitory activity, with MIC and MBC values of 20 μg/μl and 140 μg/μl, respectively.Conclusions.In vitromicrobiological tests demonstrate that the hydroalcoholic extracts of pomegranate juice and peel are able to contrast the main cariogenic bacteria involved in tooth decay. Although being preliminary data, our results suggest that pomegranate polyphenolic compounds could represent a good adjuvant for the prevention and treatment of dental caries.

scholarly journals Inhibitory Effects of Lactoferrin on Growth and Biofilm Formation of Porphyromonas gingivalis and Prevotella intermedia

2009 ◽  
Vol 53 (8) ◽  
pp. 3308-3316 ◽  
Author(s):  
Hiroyuki Wakabayashi ◽  
Koji Yamauchi ◽  
Tetsuo Kobayashi ◽  
Tomoko Yaeshima ◽  
Keiji Iwatsuki ◽  
...  
Keyword(s):  
Porphyromonas Gingivalis ◽  
Biofilm Formation ◽  
Periodontal Diseases ◽  
Antimicrobial Protein ◽  
Antibiofilm Activity ◽  
Prevotella Intermedia ◽  
Periodontopathic Bacteria ◽  
Oral Pathogens ◽  
In Vitro Effects

ABSTRACT Lactoferrin (LF) is an iron-binding antimicrobial protein present in saliva and gingival crevicular fluids, and it is possibly associated with host defense against oral pathogens, including periodontopathic bacteria. In the present study, we evaluated the in vitro effects of LF-related agents on the growth and biofilm formation of two periodontopathic bacteria, Porphyromonas gingivalis and Prevotella intermedia, which reside as biofilms in the subgingival plaque. The planktonic growth of P. gingivalis and P. intermedia was suppressed for up to 5 h by incubation with ≥130 μg/ml of human LF (hLF), iron-free and iron-saturated bovine LF (apo-bLF and holo-bLF, respectively), and ≥6 μg/ml of bLF-derived antimicrobial peptide lactoferricin B (LFcin B); but those effects were weak after 8 h. The biofilm formation of P. gingivalis and P. intermedia over 24 h was effectively inhibited by lower concentrations (≥8 μg/ml) of various iron-bound forms (the apo, native, and holo forms) of bLF and hLF but not LFcin B. A preformed biofilm of P. gingivalis and P. intermedia was also reduced by incubation with various iron-bound bLFs, hLF, and LFcin B for 5 h. In an examination of the effectiveness of native bLF when it was used in combination with four antibiotics, it was found that treatment with ciprofloxacin, clarithromycin, and minocycline in combination with native bLF for 24 h reduced the amount of a preformed biofilm of P. gingivalis compared with the level of reduction achieved with each agent alone. These results demonstrate the antibiofilm activity of LF with lower iron dependency against P. gingivalis and P. intermedia and the potential usefulness of LF for the prevention and treatment of periodontal diseases and as adjunct therapy for periodontal diseases.

scholarly journals Mahkota Dewa (God’s Crown) Fruit Extract Inhibits the Formation of Periodontal Pathogen Biofilms in vitro

2019 ◽  
Vol 2 (2) ◽  
pp. 57 ◽  
Author(s):  
Diajeng Celia Radita ◽  
Armelia Sari Widyarman
Keyword(s):  
Brain Heart Infusion ◽  
Positive Control ◽  
Negative Control ◽  
Periodontal Pathogen ◽  
Bacterial Strains ◽  
Fruit Extract ◽  
Fruit Extracts ◽  
Level Of Significance ◽  
Phosphate Buffered Saline

Introduction: Mahkota dewa (Phaleria macrocarpa) is an Indonesian fruit that contains antibacterial compounds, such as flavonoids, saponins, and tannins; it has been used as an alternative treatment for controlling infection. Objectives: This study aimed to examine the effect of mahkota dewa fruit extract on the formation of Porphyromonas gingivalis (P. gingivalis), Aggregatibacter actinomycetemcomitans (A. actinomycetemcomitans), and Treponema denticola (T. denticola) biofilms in vitro. Methods: God’s crown fruit was extracted using the maceration technique, and then diluted into different concentrations (25%, 12.5%, 6.25%, 3.125%, and 1.56%) using phosphate buffered saline (PBS). P. gingivalis ATCC-33277, A. actinomycetemcomitans ATCC-29522, or T. denticola ATCC-35405 were cultured in brain heart infusion (BHI) broth, 24h (anaerobic-condition), and then each type of bacteria (108CFU/mL) was distributed into a 96-well microplate to form a biofilm. Subsequently, the fruit extracts were distributed into the biofilm-containing well plates and incubated for 1h, 6h, and 24h. A biofilm without the fruit extract and chlorhexidine-gluconate (0.2%) was used as the negative and positive control, respectively. Crystal violet (0.5%w/v) was used to determine the density of the remaining biofilm using a microplate spectrophotometer (600 nm). Data were statistically analyzed using one-way ANOVA, and p <0.05 was set as the level of significance. Results: The mahkota dewa fruit extracts significantly inhibited the formation of a biofilm for all three bacterial strains at all concentrations and for each incubation time (p <0.05) based on optical density (OD)±SD.  The best concentration of fruit extract to inhibit biofilm formation was 25% for P. gingivalis (OD=0.19±0.06), 12.5% for A. actinomycetemcomitans (OD=0.14 ± 0.16), and 25% for T. denticola (OD=1.17±0.19) in comparison to the biofilm mass of the negative control, which was 1.67±0.06, 1.17±0.34, 2.66±0.38 for P. gingivalis, A. actinomycetemcomitans, and T. denticola, respectively. Conclusion: Based on these results, mahkota dewa fruit extract can inhibit the formation of biofilm on P. gingivalis, A. actinomycetemcomitans, and T. denticola, and it may potentially be used to prevent the infection associated with periodontal disease.

scholarly journals Indonesian Strain of Lactobacillus reuteri Probiotic Reduces the Initial Biofilm Colonization

2020 ◽  
Vol 14 (1) ◽  
pp. 544-553
Author(s):  
Armelia Sari Widyarman ◽  
Triska Ramajayanti ◽  
Citra Fragrantia Theodorea
Keyword(s):  
Streptococcus Mutans ◽  
Biofilm Formation ◽  
Lactobacillus Reuteri ◽  
Positive Control ◽  
Negative Control ◽  
Microbial Biofilms ◽  
Incubation Periods ◽  
Negative Controls ◽  
Initial Biofilm

Background: The benefits of probiotics for human health have long been proven. Probiotic Lactobacillus reuteri, can produce a beneficial broad-spectrum antibacterial compound called reuterin by metabolizing glycerol. Objective: The aim of the study was to investigate the effect of the Indonesian strain of L. reuteri LC382415 on mono- and dual-species Streptococcus mutans and Streptococcus sanguinis biofilms in vitro. Methods: Streptococcus mutans and S. sanguinis were cultured in BHI broth. Lactobacillus reuteri LC382415 was inoculated on MRS agar. The different concentrations effect of L. reuteri (1×104, 1×106, and 1×108 CFU/mL) with and without glycerol supplementation on microbial biofilms were examined using a biofilm assay after incubation for 1,3,6, and 24-h. The biofilm mass optical density was measured with a microplate spectrophotometer at 490 nm. Chlorhexidine gluconate (0.2%) was used as a positive control, and wells without treatment were used as negative controls. Results: A significant reduction in mono- and dual-species S. mutans and S. sanguinis biofilm formation was observed after treatment with all concentrations of L.reuteri and after all incubation periods (p<0.05) with or without glycerol supplementation. The concentration of 1×104 CFU/mL after 3-h incubation was the most effective in inhibiting biofilm formation, with 87.8% S. mutans, 95.9% S. sanguinis, and 80.4% dual-species biofilm reduction compared to the negative control (p<0.05). Conclusion: The Indonesian strain of L. reuteri effectively reduces mono- and dual-species S.mutans and S. sanguinis biofilms. This suggests that it may be useful in preventing biofilm formation in oral cavities. Future studies on the mechanism of action of this active component are warranted.

scholarly journals Control of Penicillium glabrum by Indigenous Antagonistic Yeast from Vineyards

Foods ◽  
2020 ◽  
Vol 9 (12) ◽  
pp. 1864
Author(s):  
Catalina M. Cabañas ◽  
Alejandro Hernández ◽  
Ana Martínez ◽  
Paula Tejero ◽  
María Vázquez-Hernández ◽  
...  
Keyword(s):  
Biofilm Formation ◽  
Food Preservation ◽  
Lytic Enzymes ◽  
Antagonist Activity ◽  
Pichia Kudriavzevii ◽  
Hanseniaspora Uvarum ◽  
Antagonistic Yeast ◽  
Negative Controls

Biocontrol is one of the most promising alternatives to synthetic fungicides for food preservation. Botrytis cinerea, Alternaria alternata, and Aspergillus section Nigri are the most concerning pathogens for grape development. However, frequently, other species, such as Penicillium glabrum in this study, are predominant in spoiled bunches. In this work, 54 native yeasts from vineyards were screened by direct confrontation in potato dextrose agar plates as antagonists against P. glabrum. Isolates of Pichia terricola, Aureobasidium pullulans, and Zygoascus meyerae were selected for their antagonist activity in vitro, plus isolates of Pichia kudriavzevii, Hormonema viticola, and Hanseniaspora uvarum were used as negative controls. However, in vivo, confrontations in wounded grapes showed disagreement with direct confrontation in vitro. P. terricola, P. kudriavzevii, H. viticola, Z. meyerae, and H. uvarum significantly reduced the incidence of P. glabrum on grapes. Nevertheless, P. terricola, H. viticola, and H. uvarum themselves spoiled the wounded grapes. Inhibitions were associated with different mechanisms such as the production of volatile organic compounds (VOCs), lytic enzymes, biofilm formation, and competition for nutrients. The isolates of P. kudriavzevii L18 (a producer of antifungal VOCs which completely inhibited the incidence of P. glabrum) and Z. meyerae L29 (with pectinase, chitinase and β-glucanase activity and biofilm formation which reduced 70% of the incidence of P. glabrum) are proposed as suitable biocontrol agents against P. glabrum.

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