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Urinary Tract Health and Antibacterial Benefits: In-Vitro

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A Comparative Evaluation of the Antimicrobial Effects of Different Mouthrinses against Oral Pathogens: An In Vitro Study

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Authors
Abu-Obaid E; Salama F; Abu-Obaid A; Alanazi F; Salem M; Auda S; Al Khadra T.
Journal
Journal of Contemporary Dental Practice. 21(5):500-508, 2020 May 01.
Abstract

AIM: To assess the antimicrobial effects of natural and semi-natural mouthrinses on isolates of Streptococcus mutans, Lactobacillus fermentum, and Lactobacillus casei obtained from the saliva samples and their reference strains. MATERIALS AND METHODS: Natural and semi-natural mouthrinses included in this study were herbal mix mouthrinse, cranberry mouthrinse, chlorhexidine digluconate mouthrinse, cranberry extract mixed with chlorhexidine digluconate mouthrinse, chlorhexidine digluconate mouthrinse with alcohol (positive control), and distilled water (negative control). The microbiological examination tests were minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC), and zone of inhibition test for the saliva isolates of S. mutans, L. fermentum, and L. casei while zone of inhibition test only for their reference strains. RESULT: Compared with distilled water, herbal mix, cranberry, cranberry mixed with chlorhexidine, chlorhexidine with alcohol (+), and chlorhexidine mouthrinses were associated with a significant increase of the zone of inhibition 34.354, 34.255, 34.219, 10.801, and 9.386, respectively. Both MIC and MBC were significantly higher in the cranberry mixed with chlorhexidine than in chlorhexidine with alcohol. The MIC and MBC of mouthrinses were significantly lower in the S. mutans and L. fermentum than in L. casei. CONCLUSION: Herbal mix and cranberry mouthrinses could be effective natural alternative to chlorhexidine mouthrinse with or without alcohol in improving oral health. CLINICAL SIGNIFICANCE: Different mouthrinses proposed in this study showed antimicrobial effects against the tested oral pathogens, and possibly the tested mouthrinses will lead for future formulation of natural or semi-natural pharmaceutical mouthrinses.

Antibacterial activities of a polyphenolic-rich extract prepared from American cranberry (Vaccinium macrocarpon) fruit pomace against Listeria spp.

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Authors
Diarra, M. S.; Hassan, Y. I.; Block, G. S.; Drover, J. C. G.; Delaquis, P.; Oomah, B. D..
Journal
LWT - Food Science and Technology; 2020. 123
Abstract

Cranberry (Vaccinium macrocarpon) fruits are known for their high polyphenolics content making them a rich source of antioxidants. These polyphenolics have been reported to promote human health and are gaining attention for their antimicrobial activities against foodborne pathogens. We investigated the antimicrobial activity of an ethanolic extract (#FC111-1) prepared from cranberry pomace against Listeria spp. Many polyphenolics were identified in this extract which could be responsible for growth-inhibitory effects against 12 Listeria strains including L. monocytogenes. The minimum inhibitory concentration (MIC) of #FC111-1 determined in cation adjusted Muller Hinton broth (CAMHB) was approximately 2 mg/mL for 11 (91.7%) of these strains. The inclusion of 2-8 mg/mL (1-4 x MIC) of #FC111-1 decreased (>3 log10) viable bacterial cells of all Listeria strains in CAMHB over a 24 h period, while dose-dependently reducing bacterial salt tolerance, bacterial bile-salt hydrolase activity, bacterial biofilm formation capacity, and increasing cell-membrane permeability. The #FC111-1 extract (0.4 and 0.8% concentrations) had no effect on L. monocytogenes survival in a cooked chicken-breast meat model, highlighting the influence of protein-rich matrices on antibacterial activity and the need to consider the role of food composition when using extracts or polyphenolics from cranberry fruits to improve food-safety

New evidences of antibacterial effects of cranberry against periodontal pathogens

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Authors
Sanchez, M. C.; Ribeiro-Vidal, H.; Bartolome, B.; Figuero, E.; Moreno-Arribas, M. V.; Sanz, M.; Herrera, D
Journal
Foods; 2020. 9(2).
Abstract

The worrying rise in antibiotic resistances emphasizes the need to seek new approaches for treating and preventing periodontal diseases. The purpose of this study was to evaluate the antibacterial and anti-biofilm activity of cranberry in a validated in vitro biofilm model. After chemical characterization of a selected phenolic-rich cranberry extract, its values for minimum inhibitory concentration and minimum bactericidal concentration were calculated for the six bacteria forming the biofilm (Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Porphyromonas gingivalis, and Aggregatibacter actinomycetemcomitans). Antibacterial activity of the cranberry extract in the formed biofilm was evaluated by assessing the reduction in bacteria viability, using quantitative polymerase chain reaction (qPCR) combined with propidium monoazide (PMA), and by confocal laser scanning microscopy (CLSM), and anti-biofilm activity by studying the inhibition of the incorporation of different bacteria species in biofilms formed in the presence of the cranberry extract, using qPCR and CLSM. In planktonic state, bacteria viability was significantly reduced by cranberry (p < 0.05). When growing in biofilms, a significant effect was observed against initial and early colonizers (S. oralis (p 0.017), A. naeslundii (p = 0.006) and V. parvula (p = 0.010)) after 30 or 60 s of exposure, while no significant effects were detected against periodontal pathogens (F. nucleatum, P. gingivalis or A. actinomycetemcomitans (p > 0.05)). Conversely, cranberry significantly (p < 0.001 in all cases) interfered with the incorporation of five of the six bacteria species during the development of 6 h-biofilms, including P. gingivalis, A. actinomycetemcomitans, and F. nucleatum. It was concluded that cranberry had a moderate antibacterial effect against periodontal pathogens in biofilms, but relevant anti-biofilm properties, by affecting bacteria adhesion in the first 6 h of development of biofilms

Comparative Susceptibility Study Against Pathogens Using Fermented Cranberry Juice and Antibiotics.

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Authors
Mantzourani I; Bontsidis CA; Plessas S; Alexopoulos A; Theodoridou E; Tsigalou C; Voidarou C; Douganiotis G; Kazakos SL; Stavropoulou E; Bezirtzoglou E.
Journal
Frontiers in Microbiology. 10:1294
Abstract

In the present study, unfermented and fermented cranberry juice in combination with the Antibiotics vancomycin and tigecycline were tested for their antimicrobial activity. Cranberry juice was fermented with a recently isolated potentially probiotic Lactobacillus paracasei K5. The tested strains selected for this purpose were Enterococcus faecalis, E. faecium, Enterobacter cloacae and Staphylococcus aureus. The methods followed were the determination of zones inhibition, Minimum Inhibitory Concentration (MIC) and Fractional Inhibitory Concentration Index (FICI). Tigecycline together with fermented juice exhibited larger Zones of Inhibition (ZOI) in strains of E. faecium (65 +/- 4.8 mm) compared to the respective ZOI with tigecycline and unfermented juice (no zone). The same outcome was also obtained with E. cloacae. Vancomycin together with fermented juice exhibited larger ZOI in strains of E. faecium (28 +/- 2.2 mm) compared to the respective ZOI with vancomycin and unfermented juice (24 +/- 2.3 mm). The lowest MIC values were recorded when tigecycline was combined with fermented cranberry juice against S. aureus strains, followed by the same combination of juice and antibiotic against E. cloacae strains. FICI revealed synergistic effects between fermented juice and tigecycline against a strain of E. faecium (A2020) and a strain of E. faecalis (A1940). Such effects were also observed in the case of fermented juice in combination with vancomycin against a strain of S. aureus (S18), as well as between fermented juice and tigecycline against E. cloacae (E1005 and E1007) strains. The results indicate that the antibacterial activity of juice fermented with the potentially probiotic L. paracasei K5 may be due to synergistic effects between some end fermentation products and the antibiotic agents examined.

Cranberry Extract with Enhanced Bactericidal Activities Against Uropathogenic Escherichia Coli within one Minute of Treatment

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Authors
Kim SA, Kim HW, Rhee MS
Journal
LWT 113:108318, https://doi.org/10.1016/j.lwt.2019.108318
Abstract

Cranberry has been widely utilized as a popular botanical dietary supplement to prevent urinary tract infection. The study aims to evaluate the enhanced bactericidal activities of cranberry against uropathogenic Escherichia coli (UPEC) by adding a small quantity of naturally derived antimicrobials. The antibacterial effect was examined with cranberry extract alone (15 and 20%), three kinds of medium-chain fatty acids alone (caprylic, capric, and lauric acid; 0.05–1.0 mM), essential oils alone (carvacrol and thymol; 0.5–1.0 mM), and cranberry extract containing medium-chain fatty acids or essential oils at 37 °C for 1 min. The survivors were remarkably reduced with cranberry extract containing any of the antimicrobials. For example, cranberry extract (15 and 20%) with 1.0 mM of each caprylic acid, lauric acid, and carvacrol resulted in the complete eradication of UPEC (7.55 log reduction). Flow cytometry analysis of UPEC cells exposed to combined treatment showed clear membrane disruption and cell death (>95% of damage). Adding antimicrobials to cranberry extract did not affect (P > 0.05) the characteristics of the cranberry extract (Color, °Brix, pH). The present method may be more acceptable to consumers, who tend to avoid products containing synthetic chemicals and prefer the use of natural agents.

Effect of Polyphenol-Rich Cranberry Extracts on Cariogenic Biofilm Properties and Microbial Composition of Polymicrobial Biofilms.

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Authors
Philip N; Bandara HMHN; Leishman SJ; Walsh LJ.
Journal
Archives of Oral Biology. 102:1-6
Abstract

OBJECTIVE: To investigate the effect of cranberry extracts on saliva-derived polymicrobial biofilms with regards to biofilm biomass, acidogenicity, exopolysaccharide (EPS)/microbial biovolumes, colony forming unit (CFU) counts, and the relative abundance of specific caries- and health-associated bacteria.METHODS: Saliva-derived polymicrobial biofilms were grown for 96 h in a cariogenic environment and treated for 2 min every 12 h over the entire biofilm growth period with 500 mug/mL cranberry extract or vehicle control. The effect of the cranberry extract on biofilm behaviour was evaluated using different assays and its influence on key cariogenic and health-associated bacterial populations was assessed with a microarray real-time quantitative PCR method.RESULTS: Cranberry-treated biofilms showed significant drops in biomass (38% reduction, P < 0.001), acidogenicity (44% reduction, P < 0.001), EPS/microbial biovolume ratios (P = 0.033), and CFU counts (51% reduction, P = 0.001). Furthermore, the cranberry extracts effected a significantly lower relative abundance of caries-associated Streptococcus sobrinus (fold change 0.004, P = 0.002) and Provotella denticola (0.002, P < 0.001), and a significantly higher relative abundance of the health-associated Streptococcus sanguinis (fold change 90.715, P = 0.001).CONCLUSIONS: The cranberry extract lowered biofilm biomass, acidogenicity, EPS/microbial biovolumes, CFU counts, and modulated a beneficial microbial ecological change in saliva-derived polymicrobial biofilms.

Inhibitory Effects of Lingonberry Extract on Oral Streptococcal Biofilm Formation and Bioactivity.

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Authors
Kokubu E; Kinoshita E; Ishihara K.
Journal
Bulletin of Tokyo Dental College. 60(1):1-9, 2019
Abstract

Phenolic compounds in fruits such as cranberries have been shown to promote a number of biological activities. The purpose of this study was to investigate the effects of polyphenolic compound-containing lingonberry extract on oral streptococci and compare them with the known anti-cariogenic activity of cranberries. Water-soluble and polyphenol-rich fractions (Fractions I and II, respectively) were isolated from cranberries and lingonberries. The effects of those fractions on the biofilm formation ability and bioactivity of Streptococcus mutans MT8148R, Streptococcus sobrinus 6715, and Streptococcus sanguinis ATCC 10556 were then evaluated. Cranberry or lingonberry Fraction II (at 0.5-1 mg/ml) significantly reduced biofilm formation by S. mutans, S. sobrinus, and S. sanguinis. In contrast, cranberry or lingonberry Fraction I (at 0.5-2 mg/ml) increased biofilm formation by S. mutans and S. sobrinus, but not by S. sanguinis. Fractions I and II (at 1-2 mg/ml) also reduced the bioactivity of S. mutans, while Fraction II (at 0.5 mg/ml) enhanced the bioactivity of all tested strains. The results revealed that lingonberries contained a larger amount of polyphenol than cranberries and that they showed almost the same level of activity against the biofilm formation ability and bioactivity of oral streptococci. This indicates that polyphenol-rich lingonberry fraction offers a promising natural food derivative for prevention of dental caries.

Polyphenol-Rich Cranberry Extracts Modulate Virulence of Streptococcus mutans-Candida albicans Biofilms Implicated in the Pathogenesis of Early Childhood Caries.

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Authors
Philip N; Leishman SJ; Bandara H; Walsh LJ.
Journal
Pediatric Dentistry. 41(1):56-62
Abstract

Purpose: The purpose of this study was to investigate the effects of polyphenol-rich cranberry extracts on dual-species Streptococcus mutans-Candida. albicans biofilms implicated in contributing to the severity of early childhood caries. Methods: S. mutans-C. albicans biofilms were grown on saliva-coated hydroxyapatite discs (s-HA) mounted on the high-throughput Amsterdam Active Attachment model. The s-HA discs were treated with the cranberry extracts/vehicle control for five minutes just before biofilm growth and subsequently, for similar exposure times, after 12 hours and 24 hours of biofilm growth. The treated 24-hour-old biofilms were then assessed for acidogenicity, metabolic activity, exopolysaccharide (EPS)/microbial biovolumes, structural organization, and colony forming unit (CFU) counts. Results: Treatment with 500 to 1,000 mug/mL of the cranberry extracts produced significant reductions in acidogenicity and metabolic activity (P<0.0001) compared to the control-treated biofilms. A significant decrease in biovolumes of the EPS (P=0.003) and microbial biofilm components (P=0.007) was also seen. Qualitative assessment of confocal biofilm images revealed that the cranberry extract disrupted biofilm structural architecture. Finally, significantly fewer S. mutans (P=0.006) and C. albicans (P=0.036) CFUs were recovered from the cranberry-treated biofilms than from the control-treated bio-films. Conclusions: Cranberry extracts inhibited cariogenic virulence properties of S. mutans-C. albicans dual-species biofilms in an in vitro model.

Proanthocyanidin Interferes with Intrinsic Antibiotic Resistance Mechanisms of Gram-Negative Bacteria

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Authors
Maisuria VB, Okshevsky M, Déziel E, and Tufenkji N
Journal
Advanced Science 6:1802333, DOI: 10.1002/advs.201802333
Abstract

Antibiotic resistance is spreading at an alarming rate among pathogenic bacteria in both medicine and agriculture. Interfering with the intrinsic resistance mechanisms displayed by pathogenic bacteria has the potential to make antibiotics more effective and decrease the spread of acquired antibiotic resistance. Here, it is demonstrated that cranberry proanthocyanidin (cPAC) prevents the evolution of resistance to tetracycline in Escherichia coli and Pseudomonas aeruginosa, rescues antibiotic efficacy against antibiotic-exposed cells, and represses biofilm formation. It is shown that cPAC has a potentiating effect, both in vitro and in vivo, on a broad range of antibiotic classes against pathogenic E. coli, Proteus mirabilis, and P. aeruginosa. Evidence that cPAC acts by repressing two antibiotic resistance mechanisms, selective membrane permeability and multidrug efflux pumps, is presented. Failure of cPAC to potentiate antibiotics against efflux pump-defective mutants demonstrates that efflux interference is essential for potentiation. The use of cPAC to potentiate antibiotics and mitigate the development of resistance could improve treatment outcomes and help combat the growing threat of antibiotic resistance

Proanthocyanidin-Chitosan Composite Nano Particles Prevent Bacterial Invasion and Colonization of Gut Epithelial Cells by Extra-Intestinal Pathogenic Escherichia Coli.

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Authors
Alfaro-Viquez E; Esquivel-Alvarado D; Madrigal-Carballo S; Krueger CG; Reed JD.
Journal
International Journal of Biological Macromolecules. 135:630-636
Abstract

Cranberry proanthocyanidin-chitosan composite nanoparticles (PAC-CHT NPs) were formulated using 2:1, 5:1, 10:1, 15:1 20:1, 25:1, and 30:1 PAC to CHT weight ratio to form round shaped particles. The PAC-CHT NPs were characterized by size, polydispersity, surface charge, morphology, and PAC content. PAC-CHT NPs bioactivity was measured by agglutination of extra-intestinal pathogenic Escherichia coli (ExPEC) and inhibition of gut epithelial cell invasion by ExPEC. Results indicate that by increasing the PAC to CHT ratio 10:1 to 30:1 formed stable nanoparticles with diameters of 122.8 to 618.7nm, a polydispersity index of approximated 0.4 to 0.5, and a zeta potential of 34.5 to 54.4mV. PAC-CHT NPs ratio 30:1 agglutinated ExPEC and decreased the ability of ExPEC to invade epithelial cells in a dose-dependent manner. PAC-CHT NPs ratio 10:1 to 30:1 form stable, round-shaped, and bioactive nanoparticles for potential applications in the treatment of ExPEC bacterial infections.