Effectiveness of pre-procedural rinsing with essential oils-based mouthrinse to reduce aerosol contamination of periodontitis patients

This research aims to evaluate the effectiveness of pre-procedural rinsing using essential oils-mouthwash (Listerine®) in reducing bioaerosol contamination in a dental clinic. Thirty (30) subjects who consisted of those receiving treatment for periodontitis problems were randomly assigned to rinse with either 20 mL of Listerine® or 20 mL of placebo as control rinse. Every subject was instructed to gargle using the rinse for 1 min. Microbial samples of environmental air and saliva were collected before and after the rinse. All samples were further analyzed for total plate counts to measure the microbial level. Rinsing using Listerine® showed significant reduction in the level of microbial load in saliva compared to the control mouthrinses. Analysis done at three defined distance intervals from the operating site showed the level of bioaerosol contamination was highest at distance nearest to the treatment point of 1 ft. Based on counts of cfu, there was higher presence of microbial contaminant in bioaerosols of the control-rinsed group compared to the test-rinsed group using Listerine®. Therefore, it can be concluded that rinsing using Listerine® was effective towards reducing the microorganisms in saliva and oral cavity in general

Distribution of Candida in the oral cavity and its differentiation based on the internally transcribed spacer (ITS) regions of rDNA

This study aimed to determine the distribution of Candida species in the oral cavity and differentiate the species based on PCR amplification, including HinfI and MspI digestion, in order to assess the effectiveness of using the rDNA region for species identification. Samples from saliva as well as palate, tongue and cheek mucosa surfaces were collected from 45 individuals, consisting of three groups: periodontal disease patients; denture-wearers; and the control group. The samples were serially diluted, spread on BHI and YPD agar plates and scored for colony-forming units (CFUs). Fifteen random candidal colonies were isolated and subjected to genomic DNA extraction, based on glass beads disruption. Four primers were used to amplify regions in the rDNA, and the ITSI-5.8S-ITSII PCR product was digested by HinfI and MspI restriction enzymes. The microbial loads on all sites of the denture-wearers were found to be significantly higher than control, while in the periodontal disease group only the microbial loads on the tongue were significantly higher than control. Meanwhile, there was no significant difference at other sites. The restriction fragment lengths of the clinical samples were compared to those of seven control species, allowing the differentiation of all seven species and the identification of 14 species from the clinical samples. The MspI restriction digest was not able to distinguish between C. albicans and C. dubliniensis, whereas the HinfI digest could not distinguish between C. tropicalis and C. parapsilosis. It was concluded that PCRRFLP of the candidal rDNA region has potential for species identification. This study demonstrates the potential use of candidal rDNA as a means for identifying Candida species, based on genotype. The results also indicate the possibility of constructing genetic probes that target specific restriction fragments in the ITSI-5.8S-ITSII region, enabling swift and precise identification of Candida species. Copyright (c) 2012 John Wiley & Sons, Ltd

Pathogenicity of Porphyromonas asaccharolytica in Adult Periodontitis of Malaysian Samples

Bacterial toxin of Porphyromonas asaccharolytica strains obtained from adult periodontitis were analysed using mouse model. Results from subcutaneous injection with whole bacterial cells of 108 and 1010 cells mL-1 failed to induce any infection. However when bacterial concentration of 1012 cells mL-1 was used, all P. asaccharolytica strains were able to induced the development of localized lesion on balb/c mice. In addition, infected mice appeared cachectic with ruffled hair. Thus, infection with the latter concentration was pathogenic and was dose dependent. The toxin was also found to be heat labile for all strains. Therefore, we conclude that the extracellularly toxin produced by P. asaccharolytica of our local isolates which were obtained from adult periodontitis, is virulent and heat labile

Influence of nicotine on the adherence of Candida albicans ATCC 14053 and Candida parapsilosis ATCC 22019 and expression of selected binding-related genes

Smoking is considered one of the main factors for development of dental plaque. Yeasts have been described to contribute to the plaque community. The aim of this study was to investigate the influence of nicotine on the growth of planktonic cells and biofilm, cell aggregation, surface hydrophobicity, cell adherence and binding-related genes expressed by Candida albicans and Candida parapsilosis in vitro. The relative number of viable fungal cells was determined based on viable plate counts. The biofilm growth was quantified using the crystal violet assay. Reverse transcription-polymerase chain reaction was used to evaluate the regulation of selected genes associated with adherence. The results indicated that nicotine enhanced the growth of both planktonic and biofilm oral fungal cells. Cell surface hydrophobicity and the expression of Hyphal Wall Protein 1 (HWP1) and agglutinin-like sequences 3 (ALS3) of C. albicans and C. parapsilosis were found to increase relative to the nicotine concentrations. The results suggest that nicotine can enhance the growth of C. albicans and C. parapsilosis in vitro and influence their adherence to the surface of microplate wells that mimic as the tooth surface

Characterization of Intracellular Toxin from Periodontopathic Prevotella intermedia Isolates in Malaysia

Periodontal disease is a global public health problem, including in Malaysia. The destruction of the periodontium has been associated with subgingival plaque microflora. Prevotella intermedia has been categorized as a periodontopathic bacteria as it contributes to the development and progression of periodontal disease. Therefore, a study on P.intermedia�s virulent properties is important for the understanding of periodontal disease. Thus, the aim of the study is to characterize the virulence effect of the intracellular toxin. In the study, intracellular toxin of P.intermedia from ninety isolates were obtained and injected subcutaneously in male balb/c mice, aged between 8-12 weeks old. The results showed that the intracellular toxin from all P.intermedia isolates were able to cause the development of localized lesions on the skin of balb/c mice when 0.1, 0.2 and 0.3 mL filtrates were used. In addition, it was found that the degree of skin lesions on the balb/c mice was dose dependent as the infective dose was found to correspond to the development of larger lesion accompanied by skin coagulative necrosis. In the second part of the study, the intracellular toxin was heated for 1, 5, 15, 30, 45 and 60 min at different temperatures of 37 and 60 C prior to the infection in mice. It was discovered that the intracellular toxin was not stable and sensitive towards high temperature and is thermolabile when the activity was terminated upon heating to 60 C as none of the injected mice developed skin lesion

Inhibitory Effect of Chlorhexidine and Clove Extract Oral Rinses in Reducing Dental Plaque Microbes

Dental plaque is a thin film of microorganisms on the tooth surfaces and contributes to the development of dental caries and periodontal disease. It has been widely known that the usage of antimicrobial oral rinses plays an important role in maintaining oral hygiene mainly by reducing the numbers of dental plaque microbes. This study has been carried out with the aim of comparing the clinical effectiveness of Chlorhexidine (CHX) (Oradex®) and commercialized clove extract (Mustika Ratu®) oral rinses in reducing dental plaque microorganisms. In addition, the aim was also to determine the duration of effects and magnitude of suppression of plaque microbes with the usage of the oral rinses. The antibacterial effects of CHX and clove extract oral rinses of dental plaque microbes were evaluated in healthy adults, which were not on any antibiotic treatment for the past 6 months. The oral rinses were tested on volunteers for a period of 30 min interval for up to 120 min. Volunteers were required to suspend normal oral hygiene habits and on the sampling day, the tooth surfaces of each volunteer were swabbed, followed by rinsing with the specified oral rinse. The tooth surfaces were swabbed again after 30, 60 and 120 min. Swab samples were serially diluted and plated on BHI agar. Phenotypic appearance and the colony forming units (CFU mL 1) were obtained. Results indicated that bacterial colony formation was significantly inhibited by both oral rinses and exhibited similar levels of antibacterial activity. However, the oral rinse containing CHX exhibited a longer duration of activity in controlling oral microbes compared to clove extract oral rinse. In conclusion, CHX has a better antimicrobial action to reduce and exhibit longer magnitude of suppression of dental plaque microbes than clove extract oral rinse

Anti-plaque effect of a synergistic combination of green tea and Salvadora persica L. against primary colonizers of dental plaque

Objective Green tea (Gt), leafs of Camellia sinensis var. assamica, is widely consumed as healthy beverage since thousands of years in Asian countries. Chewing sticks (miswak) of Salvadora persica L. (Sp) are traditionally used as natural brush to ensure oral health in developing countries. Both Gt and Sp extracts were reported to have anti-bacterial activity against many dental plaque bacteria. However, their combination has never been tested to have anti-bacterial and anti-adherence effect against primary dental plaque colonizers, playing an initial role in the dental plaque development, which was investigated in this study. Methods Two-fold serial micro-dilution method was used to measure minimal inhibitory concentration (MIC) of aqueous extracts of Gt, Sp and their combinations. Adsorption to hexadecane was used to determine the cell surface hydrophobicity (CSH) of bacterial cells. Glass beads were used to mimic the hard tissue surfaces, and were coated with saliva to develop experimental pellicles for the adhesion of the primary colonizing bacteria. Results Gt aqueous extracts exhibited better anti-plaque effect than Sp aqueous extracts. Their combination, equivalent to 1/4 and 1/2 of MIC values of Gt and Sp extracts respectively, showed synergistic anti-plaque properties with fractional inhibitory concentration (FIC) equal to 0.75. This combination was found to significantly reduce CSH (p < 0.05) and lower the adherence ability (p < 0.003) towards experimental pellicles. Conclusion Combination between Gt and Sp aqueous extracts exhibited synergistic anti-plaque activity, and could be used as a useful active agent to produce oral health care products

Evaluation of Salvadora persica L. and green tea anti-plaque effect: a randomized controlled crossover clinical trial

Background: In the author's earlier in vitro investigation, a combination of 0.25 mg/ml green tea and 7.82 mg/ml Salvadora persica L. aqueous extracts was found to exhibit significant synergistic anti-bacterial and anti-adherence effects against primary plaque colonizers biofilm. A clinical trial was needed to support these preliminary in vitro results and to investigate its efficacy as a mouthwash in the control of dental plaque. Methods: A 24 h plaque re-growth, double-blinded, randomized crossover trial was carried out. Participants (n = 14) randomly rinsed with test formulation, 0.12% chlorhexidine (control) and placebo mouthwashes for 24 h. A week before the trial, all participants received scaling, polishing and oral hygiene education. On the trial day, the participants received polishing at baseline and rinsed with 15 ml of randomly allocated mouthwash twice daily without oral hygiene measures. After 24 h, plaque index was scored and then the participants entered a 6-days washout period with regular oral hygiene measures. The same protocol was repeated for the next 2 mouthwashes. Results: The results were expressed as mean (±SD) plaque index. The test mouthwash (0.931 ± 0.372) significantly reduced plaque accumulation when compared with placebo (1.440 ± 0.498, p 0.0167). Conclusions: The test mouthwash has an anti-plaque effect for a 24 h period. Longer-term clinical studies are highly encouraged to investigate its anti-plaque effect for longer periods. Trial registration: This study was registered in ClinicalTrials.gov as NCT02624336in December 3, 2015

Isolation and Antibiotic Resistance of Microorganisms from Toothbrush

This study was carried out to investigate the presence of contaminating microorganisms on regularly used toothbrush and to determine the antibiotics resistance of the isolated microbes towards selective antimicrobial agents. A few types of different species were recovered from the toothbrush, which include Pseudomonas sp., Lactobacillus sp., Leuconostoc sp., Aerococcus sp. and Staphylococcus sp. These microbes adhered to the toothbrush bristles and could be acquired from dust, skin, water or even the water pipeline system. All isolated species were tested against ampicillin, kanamycin, sulfonamide and polymyxin-B. As high as 67.5% of isolates were resistant towards ampicillin and 47.5% towards kanamycin. Besides these two antibiotics, 35.0 and 17.5% isolates were found to be resistant towards sulfonamide and polymyxin-B, respectively. In addition, all species were shown to have multiple resistance towards various antibiotics tested and all isolated species were resistant to at least two different antibiotics. Thus, these multiple antimicrobial resistance ability could be transferred from the microbes that contaminate the toothbrush to human through the prolonged usage of the same toothbrush