Caries Pathogen Research Articles

Effects of Ag/ZnO nanocomposite at sub-minimum inhibitory concentrations on virulence factors of Streptococcus mutans

ObjectiveStreptococcus mutans (S. mutans), the main pathogen of dental caries, could be well killed by Ag/ZnO nanocomposite. However, effects on virulence factors remain to be elucidated. This study investigated effects of Ag/ZnO at sub-minimum inhibitory concentrations (sub-MICs) on virulence factors of S. mutans and related genes expressions. DesignEffects of Ag/ZnO on the growth of S. mutans was investigated by growth curves and MTT staining method. The influence of Ag/ZnO at sub-MICs on biofilm formation was measured by the crystal violet staining method and observed by a scanning electron microscopy. Adherence, cell-surface hydrophobicity, acidogenicity and extracellular polysaccharides (EPS) of S. mutans after treatment by Ag/ZnO at sub-MICs were also investigated. Virulence factors related genes expressions after treated by Ag/ZnO at 1/2 MIC was conducted by the quantitative real-time PCR (qRT-PCR) method. ResultsSub-MICs of Ag/ZnO exhibited a dose-dependent inhibition on the virulence factors of S. mutans. Specially, Ag/ZnO at 1/2 MIC decreased 69.00 % biofilm formation, 31.78 % sucrose-independent and 48.08 % sucrose-dependent adherence, 69.44 % cell-surface hydrophobicity and 72.45 % water-soluble and 90.60 % water-insoluble EPS. Furthermore, the expression of virulence factors related genes was significantly suppressed by Ag/ZnO at 1/2 MIC. ConclusionsAg/ZnO at sub-MICs inhibited multiple virulence factors of S. mutans through downregulating the related genes. Ag/ZnO nanocomposite could be used for prevention of dental caries at low dosage.

The defeat of wheat varieties by the pathogen Tilletia caries (DC) Tul.

Goal. To determine the degree of resistance of collection samples of winter wheat to the pathogen of the common bunt Tilletia caries in the conditions of the Right forest-steppe of Ukraine.
 Research Methods. Field. The studies were conducted on the sites of the experimental farm «Glevakha» Vasilkovsky district of Kiev region in 2015—2017. The infectious background of the pathogen of solid bunt was created according to the method of Krivchenko V.I. Sustainability was assessed by counting the number of healthy and diseased ears. The results were differentiated in points on a 9-point scale.
 Results. Against an artificial infectious background of the pathogen of hard bunt, the stability of 115 winter wheat cultivars was evaluated. The collection was obtained from the National Center for Plant Genetic Resources of the Institute of Plant Production and included varietal samples from 15 countries: Ukraine, Russia, Moldova, Romania, Austria, Germany, Slovakia, Lithuania, Kazakhstan, Iran, Georgia, Hungary, Bulgaria and the USA. Among the varieties of winter wheat of domestic selection, highly resistant (score 9—8) to the causative agent of bunt was not found, resistance (score 7—6) was shown by the varieties Niva and Syaivo (Ukraine). Varieties of foreign selection that showed high resistance (score 9—8) — Galina, Nemchinovskaya 57 (Russia), F 02065G5-21, F 94578G3-1 / BUCUR // DELABRAD and Miranda (Romania), resistant (score 7—6) were — Course (Russia), Noroc (Romania) and MV-Toldi (Hungary).
 Conclusions. Varieties of winter wheat Niva, Syaivo, Hvulya, Veteran, L 59, Schedrost and Pobeda of Ukrainian selection, Galina, Nemchinovskaya 57, Course (Russia), F 02065G5-21, F 94578G3-1 / BUCUR // DELABRAD, Miranda, Noroc (Romania ), MV-Toldi (Hungary) in the years of research showed a high resistance to the causative agent of bunt T. caries on an artificial infectious background. All of them can be used in the selection of new productive, disease-resistant varieties.

Specific strains of Streptococcus mutans, a pathogen of dental caries, in the tonsils, are associated with IgA nephropathy

Streptococcus mutans is known to be a major causative agent of dental caries, and strains expressing the cell surface collagen-binding Cnm protein contribute to the development of several systemic diseases. A relationship between tonsillar immunity and glomerulonephritis has been recognized in IgA nephropathy (IgAN), and specific pathogens may have effects on tonsillar immunity (mucosal immunity). Here, we present findings showing a relationship between the presence of Cnm-positive S. mutans strains in the tonsils of IgAN patients and IgAN condition/pathogenesis. Analyses of tonsillar specimens obtained from patients with IgAN (n = 61) and chronic tonsillitis (controls; n = 40) showed that the Cnm protein-positive rate was significantly higher in IgAN patients. Among IgAN patients, the tonsillar Cnm-positive group (n = 15) had a significantly higher proportion of patients with high urinary protein (>1.5 g/gCr) and lower serum albumin level than the Cnm-negative group (n = 46). Additionally, Cnm protein and CD68, a common human macrophage marker, were shown to be merged in the tonsils of IgAN patients. These findings suggest that Cnm-positive S. mutans strains in the tonsils may be associated with severe IgAN.

Effects of the Ethanolic Extracts of Guava Leaves, Licorice Roots and Cloves on the Cariogenic Properties of Streptococcus mutans

Introduction: Streptococcus mutans is a major causative pathogen of dental caries. This study aimed to investigate the effects of the ethanolic extracts of Psidium guajava, Glycyrrhiza glabra and Syzygium aromaticum against cariogenic properties of Streptococcus mutans. These medicinal plants have been used traditionally as the ingredients in the herbal mouthwash for the management of dental caries. Methods: The 95% ethanolic extracts of these medicinal plants were prepared and their phytochemical contents were analyzed by using thin layer chromatography-densitometry. Antibacterial activities of the extracts against S. mutans were evaluated according to broth dilution method. The effects of the extracts against cariogenic properties of S. mutans, which are surface adherence, biofilm formation and acid production, were also investigated. Results: The biological active constituents, quercetin, glycyrrhizic acid and eugenol, were found in P. guajava, G. glabra and S. aromaticum, respectively. The three extracts possessed anti-bacterial activity against S. mutans with the MICs of 1.562, 0.195 and 1.562 mg/mL, respectively. The extracts at every concentration tested (at lower than their MICs) significantly inhibited cariogenic properties of S. mutans, which included inhibitions against surface adherence, biofilm formation and acid production. Conclusion: The ethanolic extracts of guava leaves, licorice roots and cloves produced anti-bacterial and anti-cariogenic activities against S. mutans. These plant extracts thus can potentially be developed as a product for the prevention of dental caries in the near future.

Antibacterial Effects of Fluoride in Streptococcus mutans Growth in Vitro

Introduction: Streptococcus mutans is a gram-positive coccus commonly found in the human oral cavity and is a pathogen of dental caries. S. mutans known to form biofilm in infective endocarditis heart-valve. Fluoride usage known to reduce the risk of dental caries. This study aims to analyze how usage of fluoride in inhibiting S. mutans growth.Methods: This study was an experimental study. Antibacterial activity test was performed to evaluate the minimum inhibitory concentration (MIC) using dilution method of sodium fluoride. The minimum bactericidal concentration (MBC) was determined by culturing from the previous dilution test into Chocolate Agar Plate.Results: MIC for sodium fluoride is 4,8 mg/ml and the MBC for sodium fluoride to S. mutans is 4,8 mg/ml. We found S. mutans growth in higher concentration than 19,2 mg/ml. Conclusion: S. mutans was inhibited in dilution test. Growth of the bacteria in higher concentration of sodium fluoride is explained with quasi-irreversible inhibtior effects of fluorida.

Chemical Composition of East Asian Invasive Knotweeds, their Cytotoxicity and Antimicrobial Efficacy Against Cariogenic Pathogens: An In-Vitro Study.

BackgroundGiant knotweeds originating from East Asia, such as Reynoutria japonica, and Reynoutria sachalinensis, and their hybrid such as Reynoutria x bohemica, are invasive plants in Europe and North America. However, R. japonica is also a traditional East Asian drug (Polygoni cuspidati rhizoma) used in Korean folk medicine to improve oral hygiene. The aim of this study was to evaluate the antibacterial activity of acetone extracts of Reynoutria species against dominant caries pathogen such as Streptococcus mutans and alternative pathogens, as well as characterize the phytochemical composition of extracts and examine their cytotoxicity.Material/MethodsUltrasonic extraction was used to obtain polyphenol-rich extracts. The extracts were characterized by HPLC-DAD-ESI-MS. To test bacterial viability, the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) against S. mutans, S. salivarius, S. sanguinis, and S. pyogenes were determined. The cytotoxicity of the extracts to human fibroblasts derived from gingiva was evaluated using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay.ResultsThe R. japonica extract had the highest bacteriostatic and bactericidal activity against pathogens causing caries, mainly dominant caries pathogen S. mutans (mean MIC 1000 μg/mL and MBC 2000 μg/mL), which was most likely associated with a higher content of stilbene aglycons and anthraquinone aglycons in the extract. Moreover, the R. japonica extract demonstrated the lowest cytotoxic effect on human fibroblasts and exhibited cytotoxic activity only at the concentration causing the death of all S. mutans.ConclusionsThe results indicate that the R. japonica acetone extract can be considered as a natural, antimicrobial agent for caries control.

Amino Sugars Modify Antagonistic Interactions between Commensal Oral Streptococci and Streptococcus mutans.

N-Acetylglucosamine (GlcNAc) and glucosamine (GlcN) enhance the competitiveness of the laboratory strain DL1 of Streptococcus gordonii against the caries pathogen Streptococcus mutans Here, we examine how amino sugars affect the interaction of five low-passage-number clinical isolates of abundant commensal streptococci with S. mutans by utilizing a dual-species biofilm model. Compared to that for glucose, growth on GlcN or GlcNAc significantly reduced the viability of S. mutans in cocultures with most commensals, shifting the proportions of species. Consistent with these results, production of H2O2 was increased in most commensals when growing on amino sugars, and inhibition of S. mutans by Streptococcus cristatus, Streptococcus oralis, or S. gordonii was enhanced by amino sugars on agar plates. All commensals except S. oralis had higher arginine deiminase activities when grown on GlcN and, in some cases, GlcNAc. In ex vivo biofilms formed using pooled cell-containing saliva (CCS), the proportions of S. mutans were drastically diminished when GlcNAc was the primary carbohydrate. Increased production of H2O2 could account in large part for the inhibitory effects of CCS biofilms. Surprisingly, amino sugars appeared to improve mutacin production by S. mutans on agar plates, suggesting that the commensals have mechanisms to actively subvert antagonism by S. mutans in cocultures. Collectively, these findings demonstrate that amino sugars can enhance the beneficial properties of low-passage-number commensal oral streptococci and highlight their potential for moderating the cariogenicity of oral biofilms.IMPORTANCE Dental caries is driven by dysbiosis of oral biofilms in which dominance by acid-producing and acid-tolerant bacteria results in loss of tooth mineral. Our previous work demonstrated the beneficial effects of amino sugars GlcNAc and GlcN in promoting the antagonistic properties of a health-associated oral bacterium, Streptococcus gordonii, in competition with the major caries pathogen Streptococcus mutans Here, we investigated 5 low-passage-number clinical isolates of the most common streptococcal species to establish how amino sugars may influence the ecology and virulence of oral biofilms. Using multiple in vitro models, including a human saliva-derived microcosm biofilm, experiments showed significant enhancement by at least one amino sugar in the ability of most of these bacteria to suppress the caries pathogen. Therefore, our findings demonstrated the mechanism of action by which amino sugars may affect human oral biofilms to promote health.

In vitro evaluation of the cariogenic potential of rebaudioside A compared to sucrose and xylitol.

The aim of the present study was to evaluate the growth and acid synthesis of cariogenic pathogens in vitro when rebaudioside A was the sole source of carbon, compared to sucrose, xylitol, and a commercial sweetener containing rebaudioside A. First, sucrose or sugar substitutes were added to human saliva and the pH was measured over 10h. Subsequently, the growth and acid synthesis efficiency of Streptococcus mutans, Streptococcus sobrinus, Streptococcus oralis, Lactobacillus rhamnosus, Lactobacillus paracasei, and Candida albicans were investigated: after adding them to minimal culture media containing only one of the carbon sources, the optical density and pH were determined over 10h. A significant pH drop only occurred in the sucrose group of human saliva whereas no differences were found between the sugar substitute groups. The individual strains of caries pathogens showed significantly lower growth rates and less acid synthesis in the sugar replacement groups, in comparison to sucrose. However, these groups also showed bacterial growth and a decline in pH. Xylitol appeared to be superior to rebaudioside A regarding the inhibition of mutans streptococci. The results for the remaining microorganisms indicate that rebaudioside A causes a similar or lower acid synthesis than xylitol. As saliva contains buffer systems, the metabolism of rebaudioside A did not seem to shift the equilibrium to demineralization. Rebaudioside A may not be suitable as a targeted Streptococcus mutans treatment, but it could contribute to a reduction of caries risk as a replacement for sucrose.

Proton-pumping F-ATPase plays an important role in Streptococcus mutans under acidic conditions

Streptococcus mutans, a bacterium mainly inhabiting the tooth surface, is a major pathogen of dental caries. The bacterium metabolizes sugars to produce acids, resulting in an acidic microenvironment in the dental plaque. Hence, S. mutans should possess a mechanism for surviving under acidic conditions. In the current study, we report the effects of inhibitors of Escherichia coli proton-pumping F-type ATPase (F-ATPase) on the activity of S. mutans enzyme, and the growth and survival of S. mutans under acidic conditions. Piceatannol, curcumin, and demethoxycurcumin strongly reduced the ATPase activity of S. mutans F-ATPase. Interestingly, these compounds inhibited the growth of S. mutans at pH 5.3 but not at pH 7.3. They also significantly reduced the colony-forming ability of S. mutans after incubation at pH 4.3, while showing essentially no effect at pH 7.3. These observations indicate that S. mutans is highly sensitive to F-ATPase inhibitors under acidic conditions and that F-ATPase plays an important role in acid tolerance of this bacterium.

A Streptococcus aquaporin acts as peroxiporin for efflux of cellular hydrogen peroxide and alleviation of oxidative stress

Aquaporins (AQPs) are transmembrane proteins widely distributed in various organisms, and they facilitate bidirectional diffusion of water and uncharged solutes. The catalase-negative bacterium Streptococcus oligofermentans produces the highest H2O2 levels reported to date, which has to be exported to avoid oxidative stress. Here, we report that a S. oligofermentans aquaporin functions as a peroxiporin facilitating bidirectional transmembrane H2O2 transport. Knockout of this aquaporin homolog, So-AqpA, reduced H2O2 export by ∼50% and increased endogenous H2O2 retention, as indicated by the cellular H2O2 reporter HyPer. Heterologous expression of So-aqpA accelerated exogenous H2O2 influx into Saccharomyces cerevisiae and Escherichia coli cells, indicating that So-AqpA acts as an H2O2-transferring aquaporin. Alanine substitution revealed Phe-40 as a key residue for So-AqpA-mediated H2O2 transport. Northern blotting, qPCR, and luciferase reporter assays disclosed that H2O2 induces a >10-fold expression of So-aqpA Super-resolution imaging showed that H2O2 treatment increases So-AqpA protein molecules per cell by 1.6- to 3-fold. Inactivation of two redox-regulatory transcriptional repressors, PerR and MntR, reduced H2O2-induced So-aqpA expression to 1.8- and 4-fold, respectively. Electrophoretic mobility shift assays determined that MntR, but not PerR, binds to the So-aqpA promoter, indicating that MntR directly regulates H2O2-induced So-aqpA expression. Importantly, So-aqpA deletion decreased oxic growth and intraspecies competition and diminished the competitive advantages of S. oligofermentans over the caries pathogen Streptococcus mutans Of note, So-aqpA orthologs with the functionally important Phe-40 are present in all streptococci. Our work has uncovered an intrinsic, H2O2-inducible bacterial peroxiporin that has a key physiological role in H2O2 detoxification in S. oligofermentans.

Voriconazole inhibits cross-kingdom interactions between Candida albicans and Actinomyces viscosus through the ergosterol pathway

Candida albicans and Actinomyces viscosus are prominent microbes associated with dental root caries. The aim of this study was to investigate the effect of C. albicans on A. viscosus biofilms and to identify the mechanisms associated with this interaction. A. viscosus and C. albicans strains (wide-type and mutants) were used to form biofilms in vitro and in vivo, which were subsequently analysed by crystal violet assay and scanning electron microscopy (SEM) to investigate the effect of C. albicans on A. viscosus growth. A viable plate count and survival curve for C. albicans mutants and A. viscosus combinations were used to identify which C. albicans pathway was crucial for cross-kingdom interactions. Voriconazole was used to block their interactions both in vitro and in vivo. SEM, fluorescence in situ hybridisation (FISH), quantitative PCR and survival curve analyses were performed to evaluate the activity of voriconazole on C. albicans and A. viscosus interactions. The biomass and virulence of mixed-species biofilms were significantly enhanced compared with the A. viscosus biofilm alone. However, this was not observed in the mixed-species biofilms with the C. albicans mutant erg11Δ/Δ in vitro and in vivo, indicating that azoles may work on the mixed-species biofilms. As expected, voriconazole can effectively reduce the biomass of mixed-species biofilms. A high concentration of voriconazole (1 µg/mL) reduced the abundance of C. albicans, whilst a low voriconazole concentration (0.25 µg/mL) blocked their interactions similar to the effect of the erg11Δ/Δ mutant. Voriconazole may be a candidate strategy to combat root caries pathogens.

Effect of Probiotics on Dental Caries and Periodontal Pathogens: An In Vitro Study

Introduction: Probiotics are microorganisms that are administered in live forms and can improve health and well being of human beings. Literature about their antimicrobial activity against oral pathogens is limited. The present study was conceptualized to ascertain the antimicrobial capabilities of common probiotics against microorganisms involved in dental caries and periodontal diseases. The aim of this article is to ascertain the action of probiotics on dental caries pathogen, Streptococcus mutans, and periodontal microbes that included Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, and Prevotella intermedia. Materials and Methods: The present study was an in vitro type of study. A. actinomycetemcomitans was cultivated in tryptic soy-serum-bacitracin-vancomycin-fluoride agar, Po. gingivalis in modified Wilkins–Chalgren medium, and Pr. intermedia in brain heart infusion agar. The samples were cultivated for 48 h in Brewer-like anaerobic jars. The first plate was impregnated with strains of the probiotic Lactobacillus acidophilus, the second plate with strain of the probiotic S. salivarius, and the third plate was kept as the control. Zones of inhibition were calculated by unaided eye employing a Vernier caliper to the nearest millimeter, including the diameter of the disk in the measurement. Results: Analysis of variance test exhibited significant findings at 24 and 48 h. Post hoc Tukey test revealed results that were statistically significant for all microorganisms but for A. actinomycetemcomitans and Pr. intermedia. Conclusion: The findings showed significant effect of probiotics on common microbes implicated in dental caries and periodontal diseases. The present study has crucial public health implications for the prevention and control of common oral diseases in populations.

Essential Roles of the sppRA Fructose-Phosphate Phosphohydrolase Operon in Carbohydrate Metabolism and Virulence Expression by Streptococcus mutans.

The dental caries pathogen Streptococcus mutans can ferment a variety of sugars to produce organic acids. Exposure of S. mutans to certain nonmetabolizable carbohydrates, such as xylitol, impairs growth and can cause cell death. Recently, the presence of a sugar-phosphate stress in S. mutans was demonstrated using a mutant lacking 1-phosphofructokinase (FruK) that accumulates fructose-1-phosphate (F-1-P). Here, we studied an operon in S. mutans, sppRA, which was highly expressed in the fruK mutant. Biochemical characterization of a recombinant SppA protein indicated that it possessed hexose-phosphate phosphohydrolase activity, with preferences for F-1-P and, to a lesser degree, fructose-6-phosphate (F-6-P). SppA activity was stimulated by Mg2+ and Mn2+ but inhibited by NaF. SppR, a DeoR family regulator, repressed the expression of the sppRA operon to minimum levels in the absence of the fructose-derived metabolite F-1-P and likely also F-6-P. The accumulation of F-1-P, as a result of growth on fructose, not only induced sppA expression, but it significantly altered biofilm maturation through increased cell lysis and enhanced extracellular DNA release. Constitutive expression of sppA, via a plasmid or by deleting sppR, greatly alleviated fructose-induced stress in a fruK mutant, enhanced resistance to xylitol, and reversed the effects of fructose on biofilm formation. Finally, by identifying three additional putative phosphatases that are capable of promoting sugar-phosphate tolerance, we show that S. mutans is capable of mounting a sugar-phosphate stress response by modulating the levels of certain glycolytic intermediates, functions that are interconnected with the ability of the organism to manifest key virulence behaviors.IMPORTANCEStreptococcus mutans is a major etiologic agent for dental caries, primarily due to its ability to form biofilms on the tooth surface and to convert carbohydrates into organic acids. We have discovered a two-gene operon in S. mutans that regulates fructose metabolism by controlling the levels of fructose-1-phosphate, a potential signaling compound that affects bacterial behaviors. With fructose becoming increasingly common and abundant in the human diet, we reveal the ways that fructose may alter bacterial development, stress tolerance, and microbial ecology in the oral cavity to promote oral diseases.

Effects of Antimicrobial Peptide GH12 on the Cariogenic Properties and Composition of a Cariogenic Multispecies Biofilm

Dental caries is a biofilm-mediated disease that occurs when acidogenic/aciduric bacteria obtain an ecological advantage over commensal species. In previous studies, the effects of the antimicrobial peptide GH12 on planktonic bacteria and monospecies biofilms were confirmed. The objectives of this study were to investigate the effects of GH12 on a cariogenic multispecies biofilm and to preliminarily explain the mechanism. In this biofilm model, Streptococcus mutans ATCC 70061 was the representative of cariogenic bacteria, while Streptococcus gordonii ATCC 35105 and Streptococcus sanguinis JCM 5708 were selected as healthy microbiota. The results showed that GH12 was more effective in suppressing S. mutans than the other two species, with lower MIC and minimal bactericidal concentration (MBC) values among diverse type strains and clinical isolated strains. Therefore, GH12, at no more than 8 mg/liter, was used to selectively suppress S. mutans in the multispecies biofilm. GH12 at 4 mg/liter and 8 mg/liter reduced the cariogenic properties of the multispecies biofilm in biofilm formation, glucan synthesis, and lactic acid production. In addition, GH12 suppressed S. mutans within the multispecies biofilm and changed the bacterial composition. Furthermore, 8 mg/liter GH12 showed a selective bactericidal impact on S. mutans, and GH12 promoted hydrogen peroxide production in S. sanguinis and S. gordonii, which improved their ecological advantages. In conclusion, GH12 inhibited the cariogenic properties and changed the composition of the multispecies biofilm through a two-part mechanism by which GH12 directly suppressed the growth of S. mutans as well as enhanced the ecological competitiveness of S. sanguinis and S. gordonii IMPORTANCE Dental caries is one of the most prevalent chronic infectious diseases worldwide, with substantial economic and quality-of-life impacts. Streptococcus mutans has been considered the principal pathogen of dental caries. To combat dental caries, an antimicrobial peptide, GH12, was designed, and its antibacterial effects on planktonic S. mutans and the monospecies biofilm were confirmed. As etiological concepts of dental caries evolved to include microecosystems, the homeostasis between pathogenic and commensal bacteria and a selective action on cariogenic virulence have increasingly become the focus. The novelty of this research was to study the effects of the antimicrobial peptides on a controlled cariogenic multispecies biofilm model. Notably, the role of an antimicrobial agent in regulating interspecific competition and composition shifts within this multispecies biofilm was investigated. With promising antibacterial and antibiofilm properties, the use of GH12 might be of importance in preventing and controlling caries and other dental infections.

Anticariogenic Activities of Derris reticulata Ethanolic Stem Extract Against Streptococcus mutans.

Streptococcus mutans is a dominant causative pathogen of dental caries, which is a major oral health problem affecting million people worldwide. Derris reticulata is a medicinal plant possessing antimicrobial activity against several Gram-positive pathogenic bacteria. None the less, its effects on growth and cariogenic properties of S. mutans has not been clearly established. This study aimed to investigate the antibacterial and anti cariogenic activities of the D. reticulata ethanolic stem extract. The TLC analysis was performed to authenticate the D. reticulata sample. Minimum inhibition concentration and minimum bactericidal concentration were determined by using broth dilution and drop plate methods, respectively. Sucrose dependent and sucrose independent-adherences, biofilm formation and glycolytic pH drop assays were performed to evaluate the anticariogenic activity. The ethanolic stem extract of D. reticulata possessed the antibacterial activity against S. mutans with the MIC and MBC of 0.875±0.250 and 1.750±0.500 mg mL-1, respectively. The extract at the lower concentrations of sub-MIC also had significant inhibitory actions against the cariogenic properties of S. mutans, including surface adherence, biofilm formation and glycolytic acid production. The D. reticulata stem extract had a substantial anticariogenic activities and thus potentially be developed as an oral health care product for dental caries prevention in the near future.

PERBANDINGAN EFEKTIFITAS JUS LIDAH BUAYA DAN KLORHEXIDIN 0.06% TERHADAP JUMLAH KOLONI STREPTOCOCCUS MUTANS SALIVA ANAK DENGAN KARIES

Background: Aloe vera contains active components such as antrhaquinones,saponin, glucomannans, and acemannans that have been known for its antimicrobial properties. In vitro study showed that Aloe vera extract can inhibit the growth of Streptococcus mutans. Studies reported that children with caries have high level of salivary S. mutans which is the main pathogen in dental caries. The gold standard mouthrinse for dental caries is chlorhexidine gluconate, but it has several side effects. The objective of this study was to compare the effect between Aloe vera juice and 0.06% chlorhexidine mouth rinse on salivary S. mutans in children with caries.Methods: This experimental research with pretest and post-test control group design conducted on 30 children (6-12 years old) with number of carious teeth >4. Subjects divided into 2 groups: Aloe vera juice group and 0.06% chlorhexidine group, which instructed to rinse their mouth with 10 ml stated mouthwash, twice a day, during 7-day period. Unstimulated saliva was collected at baseline and after 7 days then inoculated on mitis salivarius bacitracin agar. S. mutans were counted and expressed in CFU/ml. Data were analyzed using paired t-test and independent t-test.Result: The result showed that a significant reduction in salivary S.mutanscounts was observed with Aloe vera juice mouth rinse and 0.06% chlorhexidine (p<0.05). The mean reduction of S. mutans counts was not statistically significantly between grup (p>0.05).Conclusion: These findings suggest that Aloe vera juice can reduce salivary S. mutans in children with caries so it can be considered as alternative mouthwash to prevent caries.

In vitro investigation of antimicrobial activities of ethnomedicinal plants against dental caries pathogens.

The study aimed to evaluate the antimicrobial activity of medicinal plant extracts against the bacterial pathogens prominent in dental caries. A total of 20 plant species (herbs, shrubs and trees) belonging to 18 genera and 15 families were documented for dental caries. Antimicrobial activity of solvent extracts and essential oil from plants were determined by zone of inhibition on the growth of Streptococcus mutans (MTCC 497) and Lactobacillus acidophilus (MTCC 10307) using the agar well diffusion method. The results of in vitro antimicrobial assay prove that methanol is more successful in the extraction of phytochemicals from plant samples than aqueous solvent, as methanol extracts show higher antimicrobial activity than aqueous extracts against both the test pathogens. Methanol extracts of Nigella sativa, Psidium guajava and Syzygium aromaticum were the most effective among all 20 plant samples and have potent inhibitory activity against both dental caries pathogens with minimum inhibitory concentration of 0.2mg mL- 1. N. sativa seed methanol extract was more effective with 22.3mm zone of inhibition at 0.2mg mL- 1 against S. mutans (MTCC 497), while L. acidophilus (MTCC 10307) was more sensitive to S. aromaticum bud methanol extract at 11.3mm zone of inhibition at concentration 0.1mg mL- 1. Essential oil extracted from plants also possesses strong antimicrobial activity for both test pathogens, with a minimum inhibitory concentration range of 0.05-0.16mg mL- 1. Syzygium aromaticum bud essential oil at 0.05mg mL- 1 was most active against S. mutans (MTCC 497). Plant extracts viewing antimicrobial activity with minimum inhibitory concentration show the efficacy of the plant products that could be considered as a good indicator of prospective plants for discovering new antimicrobial agents against dental caries pathogens. The findings of this study provide a lead to further polyherbal formulations for the treatment of dental caries malaise.

Inhibitory effect of surface pre-reacted glass-ionomer (S-PRG) eluate against adhesion and colonization by Streptococcus mutans

Surface Pre-reacted Glass-ionomer (S-PRG) filler is a bioactive filler produced by PRG technology, which has been applied to various dental materials. A S-PRG filler can release multiple ions from a glass-ionomer phase formed in the filler. In the present study, detailed inhibitory effects induced by S-PRG eluate (prepared with S-PRG filler) against Streptococcus mutans, a major pathogen of dental caries, were investigated. S-PRG eluate effectively inhibited S. mutans growth especially in the bacterium before the logarithmic growth phase. Microarray analysis was performed to identify changes in S. mutans gene expression in the presence of the S-PRG eluate. The S-PRG eluate prominently downregulated operons related to S. mutans sugar metabolism, such as the pdh operon encoding the pyruvate dehydrogenase complex and the glg operon encoding a putative glycogen synthase. The S-PRG eluate inhibited several in vitro properties of S. mutans relative to the development of dental caries especially prior to active growth. These results suggest that the S-PRG eluate may effectively inhibit the bacterial growth of S. mutans following downregulation of operons involved in sugar metabolism resulting in attenuation of the cariogenicity of S. mutans, especially before the active growth phase.

Campylobacter rectus in the Oral Cavity Correlates with Proteinuria in Immunoglobulin A Nephropathy Patients

Background: Periodontitis-related pathogens, such as Campylobacter or Treponema species, have recently been shown to be associated with immunoglobulin A nephropathy (IgAN). Some strains of Streptococcus mutans, a major pathogen of dental caries, harbour the cnm gene that encodes a collagen-binding protein (Cnm). This has also been demonstrated to be associated with urinary protein levels in IgAN patients. Objectives: The purpose of the present study was to analyse the association of IgAN with C. rectus, Treponema denticola and cnm-positive S. mutans in the oral cavity of humans. Methods: The presence of C. rectus, T. denticola and cnm-positive S. mutans strains in saliva samples of 117 IgAN patients and 56 healthy controls was evaluated by PCR, and the subjects’ clinical parameters were analysed. Results: C. rectus was significantly more prevalent in the IgAN group than in the control group (p < 0.05). The C. rectus-positive group was significantly associated with proteinuria in the IgAN group (p < 0.05). In addition, the C. rectus-positive and cnm-positive S. mutans group was shown to be more closely associated with urinary protein levels than the other groups (p < 0.0083). Conclusion: Our results suggest that harbouring C. rectus in the oral cavity could be associated with proteinuria in IgAN patients.

Purification and Characterization of a Biofilm-Degradable Dextranase from a Marine Bacterium.

This study evaluated the ability of a dextranase from a marine bacterium Catenovulum sp. (Cadex) to impede formation of Streptococcus mutans biofilms, a primary pathogen of dental caries, one of the most common human infectious diseases. Cadex was purified 29.6-fold and had a specific activity of 2309 U/mg protein and molecular weight of 75 kDa. Cadex showed maximum activity at pH 8.0 and 40 °C and was stable at temperatures under 30 °C and at pH ranging from 5.0 to 11.0. A metal ion and chemical dependency study showed that Mn2+ and Sr2+ exerted positive effects on Cadex, whereas Cu2+, Fe3+, Zn2+, Cd2+, Ni2+, and Co2+ functioned as inhibitors. Several teeth rinsing product reagents, including carboxybenzene, ethanol, sodium fluoride, and xylitol were found to have no effects on Cadex activity. A substrate specificity study showed that Cadex specifically cleaved the α-1,6 glycosidic bond. Thin layer chromatogram and high-performance liquid chromatography indicated that the main hydrolysis products were isomaltoogligosaccharides. Crystal violet staining and scanning electron microscopy showed that Cadex impeded the formation of S. mutans biofilm to some extent. In conclusion, Cadex from a marine bacterium was shown to be an alkaline and cold-adapted endo-type dextranase suitable for development of a novel marine agent for the treatment of dental caries.