Effect of commercial herbal toothpastes and mouth rinses on the prevention of enamel demineralization using a microcosm biofilm model.

This work evaluated the effects of commercial toothpastes and mouth rinses containing natural/herbal agents on biofilm viability, extracellular polysaccharide (EPS) production and on enamel demineralization in vitro. Microcosm biofilm was produced on bovine enamel for 5 days and treated daily with: Orgânico natural® (toothpaste/mouth rinse), Boni Natural Menta & Malaleuca® (toothpaste/mouth rinse), Propolis & Myrrh® (toothpaste), Colgate Total 12 Clean Mint® (toothpaste, positive control), Malvatricin® Plus (mouth rinse), PerioGard® (mouth rinse, positive control) or PBS (negative control). Tom's Propolis & Myrrh® and Colgate Total 12® toothpastes and Malvatricin® Plus and PerioGard® mouth rinses significantly reduced biofilm viability (p < 0.05). Only PerioGard® had significant effects on biofilm thickness and EPS. Despite the indication that Tom's Propolis & Myrrh® significantly reduced lesion depth, only Colgate Total 12® significantly reduced mineral loss. Malvatricin® Plus significantly reduced mineral loss and lesion depth, as did PerioGard®. Some herbal products, Malvatricin® Plus and Tom's Propolis & Myrrh®, showed anticaries effects.

Protective Effect of 4% Titanium Tetrafluoride Varnish on Dentin Demineralization Using a Microcosm Biofilm Model.

This study evaluated the effect of titanium tetrafluoride (TiF4) varnish on the development of dentin carious lesions. Bovine root dentin samples were treated for 6 h with: (A) 4% TiF4 varnish (2.45% F); (B) 5.42% sodium fluoride (NaF) varnish (2.45% F); (C) 2% chlorhexidine (CHX) gel - positive control; (D) placebo varnish; or (E) untreated - negative control (n = 4 × biological triplicate, n = 12). Treated dentin samples were exposed to human saliva mixed with McBain saliva (1:50) for the first 8 h in 24-well plates. Thereafter, the medium was removed, and McBain saliva containing 0.2% sucrose was applied for 16 h. From days 2 to 5, McBain saliva with sucrose was replaced daily (37°C, 5% CO2). The demineralization was measured using transverse microradiography, while the effect on biofilm was analyzed using viability, extracellular polysaccharide (EPS), and lactic acid production assays. The data were statistically analyzed (p < 0.05). All treatments (fluorides and CHX) significantly reduced the biofilm viability compared to placebo varnish and negative control. However, none of them was able to reduce the colony-forming unit counting for total microorganism, total streptococci, and Streptococcus mutans. NaF significantly reduced the number of Lactobacillus sp. compared to negative control. No effect was seen on lactic acid production neither on EPS synthesis, except that CHX significantly reduced the amount of insoluble EPS. Both fluorides were able to reduce dentin demineralization compared to placebo varnish and negative control; TiF4 had a better effect in reducing mineral loss and lesion depth than NaF. Therefore, TiF4 varnish has the best protective effect on dentin carious lesion formation using this model.

Effect of a mouthrinse containing Malva sylvestris on the viability and activity of microcosm biofilm and on enamel demineralization compared to known antimicrobials mouthrinses.

The purpose of this study was to evaluate the antimicrobial (anti-biofilm) and anti-caries (enamel demineralization prevention) effects of Malva sylvestris (Malvatricin® Plus) compared with known antimicrobial mouthrinses. Microcosm biofilm was produced on enamel, using inoculum from pooled human saliva mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated with mouthrinses for 1 min day-1. Oral-B® Complete, Listerine® Zero and Malvatricin® Plus had the greatest effect on the reduction of biofilm viability (p < 0.0001). On the other hand, lactic acid production was reduced significantly with PerioGard®, Noplak® Max and Listerine® Zero compared with the control (p < 0.0001). No significant differences were found among the mouthrinses with respect to the colony-forming unit counting (total microorganisms, total streptococci, mutans streptococci and lactobacilli) and extracellular polysaccharide production. Enamel demineralization was reduced significantly with PerioGard®, Noplak® Max and Malvatricin® Plus compared with the control (p < 0.0001). Malva sylvestris has a comparable anti-caries effect to chlorhexidine mouthrinses.

Effect of oral antimicrobial mouthrinses containing alcohol on viability of Streptococcus mutans and microcosm biofilm and on the prevention of enamel caries lesions.

PURPOSE: To evaluate the effect of PerioGard, Listerine, Noplak, Malvatricin and Cepacol commercial mouthrinses containing alcohol on the viability of Streptococcus mutans strain and of a microcosm biofilm and on the prevention of enamel demineralization. METHODS: The minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were determined against S. mutans (ATCC 25175). Microcosm biofilm formed from human saliva mixed with McBain saliva was created on bovine enamel for 5 days. From the 2nd to the 5th day, the enamel samples were exposed to McBain with 0.2% sucrose and to the mouthrinses (1 x 60 seconds). The biofilm viability was determined by fluorescence and the enamel demineralization by TMR. RESULTS: The lowest MIC and MBC values were observed for Cepacol, while the highest values were found for Listerine. The mouthrinses significantly increased the number of dead bacteria in biofilm, varying from 38.0± 11.2% (Noplak) to 58.5± 13.9% (Listerine), compared to control (12.7± 10.6%), except Periogard (30.1± 12.4%). All mouthrinses reduced mineral loss (P< 0.0001), but only Noplak and Cepacol were able to significantly reduce lesion depth. Cepacol and Noplak presented the best anti-caries effect under this experimental model. CLINICAL SIGNIFICANCE: This study shows that the anti-caries potential may vary between the commercial mouthrinses, which should be taken into account for their prescription.

Antimicrobial and anti-caries effects of a novel cystatin from sugarcane on saliva-derived multi-species biofilms.

This study evaluated the antimicrobial (anti-biofilm) and anti-caries (enamel demineralization prevention) effects of a new cystatin derived from sugarcane (CaneCPI-5). Microcosm biofilm was produced on bovine enamel specimens (4 x 4 mm; n=48) from a mixture of human saliva and McBain saliva at the first 8 h. From this moment until the end of the experiment, the enamel specimens were exposed to lsaMcBain saliva containing 0.2% sucrose and, once a day, they were treated with the test solutions for 1 min. This treatment was performed for 5 days. The solutions evaluated were: PBS (negative control), 0.12% chlorhexidine (positive control), 0.1 mg/ml CaneCPI-5 and 1.0 mg/ml CaneCPI-5. The biofilm viability was determined by fluorescence using confocal microscopy and the enamel demineralization was quantified using transverse microradiography (TMR). The data were analyzed by ANOVA/Tukey or Kruskal-Wallis/Dunn tests for biofilm and enamel, respectively (p<0.05). With respect to the antimicrobial effect, all treatment solutions significantly reduced the biofilm viability compared with PBS. The best antimicrobial effect was found for 1.0 mg/ml CaneCPI-5 (82.37±10.01% dead bacteria) that significantly differed from 0.12% chlorhexidine (73.13±15.07% dead bacteria). For the anti-caries effect, only 0.12% chlorhexidine (ΔZ: 2610, 1683-4343) performed significantly better than PBS (ΔZ: 8030, 7213-9115), but 0.12% chlorhexidine did not significantly differ from 0.1 mg/ml Cane-CPI-5. Under this experimental model, CaneCPI-5 significantly reduced the biofilm viability, but this effect was not reflected on its anti-caries potential.

Antibiofilm and anti-caries effects of an experimental mouth rinse containing Matricaria chamomilla L. extract under microcosm biofilm on enamel.

OBJECTIVE: This study evaluated the antibiofilm and anti-caries effects of an experimental mouth rinse containing aqueous extract of Matricaria chamomilla L. METHODS: Microcosm biofilm was produced on bovine enamel, from pooled human saliva mixed with McBain saliva, under 0.2 % sucrose exposure, for 5 days. The biofilm was daily treated using (1 mL/1 min): Vochysia tucanorum Mart. (2.5 mg/mL); Myrcia bella Cambess. (1.25 mg/mL); Matricaria chamomilla L. (20 mg/mL); Malva sylvestris (Malvatricin® Plus-Daudt); 0.12 % Chlorhexidine (PerioGard®-Palmolive, Positive control) and PBS (Negative control). The % dead bacteria, biofilm thickness, EPS biovolume, lactic acid concentration, the CFU counting (total microorganisms, Lactobacillus sp., total streptococci and Streptococcus mutans/S. sobrinus) were determined. Enamel demineralization was measured by TMR. RESULTS: All mouth rinses induced bacterial death compared to PBS (p < 0.0001). The biofilm thickness varied from 12 ±â€¯2 µm (chlorhexidine) to 18 ±â€¯2 µm (V. tucanorum) (ANOVA/Tukey, p < 0.0001). The EPS biovolume varied from 7(4)% (chlorhexidine) to 30(20)% (PBS) (Kruskal-Wallis/Dunn, p < 0.0001). The lactic acid production was reduced by M. sylvestris (1.1 ±â€¯0.2 g/L) and chlorhexidine (0.6 ±â€¯0.2 g/L) compared to PBS (2.6 ±â€¯1.3 g/L) (ANOVA, p < 0.0001). Malva sylvestris and chlorhexidine showed significant low CFU for total microorganisms, Lactobacillus sp. and total streptococci. Only chlorhexidine significantly reduced S. mutans/S. sobrinus. CFUs for total streptococci and Lactobacillus sp, were also significantly reduced by M. chamomilla L. Malva sylvestris (63.4 % of mineral loss reduction), chlorhexidine (47.4 %) and M. chamomilla L. (39.4 %) significantly reduced enamel demineralization compared to PBS (ANOVA/Tukey, p < 0.0001). CONCLUSION: M. chamomilla L. has lower antibiofilm action, but comparable anti-caries effect to those found for chlorhexidine, under this model. CLINICAL RELEVANCE: This study shows that the antibiofilm and anti-caries potential may vary between the commercial and experimental mouth rinses containing natural agents, with promising results for those containing Matricaria chamomilla L. and Malva Sylvestris.

Comparison between static and semi-dynamic models for microcosm biofilm formation on dentin.

OBJECTIVE: Microcosm biofilm has been applied to induce carious lesions in dentin. However, no study has been done to compare the impact of the type of model for providing nutrients to microcosm biofilm formation on dentin. This study compared the performance of two kinds of models (static and semi-dynamic) on the biofilm formation and the development of dentin carious lesions. MATERIAL AND METHODS: In both models, biofilm was produced using inoculum from pooled human saliva mixed with McBain saliva for the first 8 h (5% CO2 and 37°C). Afterwards, for the static model, the samples were placed in 24-wells microplate containing McBain saliva with 0.2% sucrose, which was replaced at 24 h. In the semi-dynamic model, the samples were submitted to artificial mouth system with continuous flow of McBain saliva with 0.2% sucrose (0.15 ml/min, 37°C) for 10 h a day (for the other 14 h, no flow was applied, similarly to the static model). After 5 days, biofilm viability was measured by fluorescence and dentin demineralization by transverse microradiography. RESULTS: Biofilm viability was significantly lower for the static compared with semi-dynamic model, while dentin demineralization was significantly higher for the first one (p<0.05). The static model was able to produce a higher number of typical subsurface lesions compared with the semi-dynamic model (p<0.05). CONCLUSIONS: The type of model (static and semi-dynamic) applied in the microcosm biofilm may have influence on it's viability and the severity/profile of dentin carious lesions.

Effect of hydroalcoholic extract of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability and activity of microcosm biofilm and on enamel demineralization.

OBJECTIVES: The aim of this study was to assess the effect of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves hydroalcoholic extracts on viability and metabolism of a microcosm biofilm and on enamel demineralization prevention. METHODOLOGY: Microcosm biofilm was produced on bovine enamel using inoculum from pooled human saliva mixed with McBain saliva, under 0.2% sucrose exposure, for 14 days. The biofilm was daily-treated with the extracts for 1 min. At the end, it was analyzed with respect to viability by fluorescence, CFU counting and extracellular polysaccharides (phenol-sulphuric acid colorimetric assay) and lactic acid (enzymatic assay) production. The demineralization was measured by TMR. The data were compared using ANOVA or Kruskal-Wallis (p<0.05). RESULTS: M. urundeuva All. at 100, 10 and 0.1 µg/mL and Q. grandiflora Mart. at 100 and 0.1 µg/mL reduced biofilm viability similarly to positive control (chlorhexidine) and significantly more than the negative-vehicle control (35% ethanol). M. urundeuva at 1000, 100 and 0.1 µg/mL were able to reduce both lactobacilli and mutans streptococci CFU counting, while Q. grandiflora (1000 and 1.0 µg/mL) significantly reduced mutans streptococci CFU counting. On the other hand, the natural extracts were unable to significantly reduce extracellular polysaccharides and lactic acid productions neither the development of enamel carious lesions. CONCLUSIONS: The extracts showed antimicrobial properties on microcosm biofilm, however, they had no effect on biofilm metabolism and caries protection.

Hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on Streptococcus mutans biofilm and tooth demineralization.

OBJECTIVES: This study evaluated the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (alone or combined) on the viability of Streptococcus mutans biofilm and on the prevention of enamel demineralization. METHODS: Strain of S. mutans (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum inhibition biofilm concentration and minimum eradication biofilm concentration were determined in order to choose the concentrations to be tested under biofilm model. S. mutans biofilm (5 × 105 CFU/ml) was produced on bovine enamel, using McBain saliva under 0.2% sucrose exposure, for 3 days. The biofilm was daily treated with the extracts for 1 min. The biofilm viability was tested by fluorescence and the enamel demineralization was measured using TMR. RESULTS: Myracrodruon urundeuva All. (Isolated or combined) at the concentrationsc ≥0.625 mg/ml was able to reduce bacteria viability, while Qualea Grandflora Mart. alone had antimicrobial effect at 5 mg/ml only (p < 0.05). On the other hand, none of the extracts were able to reduce enamel demineralization. CONCLUSIONS: The hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves (isolated or combined) have antimicrobial action; however, they do not prevent enamel caries under S. mutans biofilm model.

Commercial antimicrobials mouthrinses on caries and periodontitis-related biofilm control: a review of literature / Enxaguatórios comerciais antimicrobianos sobre o controle do biofilme relacionado à cárie dentária e à doença periodontal­ uma revisão de literatura

Esta revisão tem como objetivo discutir o potencial antimicrobiano de diferentes enxaguatórios bucais em relação ao controle da cárie dentária e doença periodontal. A pesquisa foi realizada usando PubMed e as seguintes palavras-chave: "agente antimicrobiano" ou "agente antiplaca", "biofilme dental" e "cárie dentária" ou "doença periodontal" ou "gengivite". Foram selecionados os estudos publicados em inglês, de 2011 a 2015, em revistas com fator de impacto maior que 0,8. Foram encontrados no total 22 artigos, 13 relacionados à cárie dentária e 9 relacionados à doença periodontal. Entre os 13 estudos envolvendo bactérias e/ou biofilme cariogênicos, 6 foram realizados in vitro, 3 in situ e 4 in vivo. Entre os 9 estudos envolvendo doença periodontal, 2 foram in vitro e 7 in vivo. Os principais agentes ativos testados foram: CHX-Clorexidina, CPC-cloreto de cetilpiridínio e OE-óleos essenciais (com álcool ou sem álcool). A CHX foi comparada ao OE em 6 estudos, mostrando superioridade em 3 estudos, similaridade em 1 estudo e inferioridade em 2 estudos. CPC mostrou menor efeito na redução da placa em comparação à CHX e ao OE. Ainda há controvérsias sobre o efeito do álcool, mas alguns estudos têm mostrado superioridade no caso de OE e CHX com álcool sobre biofilmes cariogênicos e periodontopatogênicos, respectivamente, quando comparados à versão sem álcool; para o CPC, não foi encontrada diferença. Mais estudos clínicos são necessários para melhor compreensão sobre mecanismo de ação e as diferenças de desempenho entre os agentes antiplaca.(AU)

This review aims to discuss the antimicrobial potential of different mouthrinses in respect to the control of dental caries and periodontal disease. The survey was conducted using PubMed and the following keywords: "antimicrobial agent" or "antiplaque agent", "dental biofilm" and "dental caries" or "periodontal disease" or "gingivitis". Only studies published in English, from 2011 to 2015, in journals with impact factor greater than 0.8, were selected. We found a total of 22 papers, 13 related to dental caries and 9 related to periodontal disease. Among the 13 studies involving cariogenic bacteria and/or biofilm, 6 were conducted in vitro, 3 in situ and 4 in vivo. Among 9 studies involving periodontal disease, 2 were in vitro and 7 in vivo. The main active agents tested were: CHX-Chlorhexidine, CPC-cetylpyridinium chloride and EO-Essential oils (alcohol/or alcohol-free). CHX was compared to EO in 6 studies, showing superiority in 3 studies, similarity in 1 study and inferiority in 2 studies. CPC has shown lower effect in plaque reduction compared to CHX and EO. There is still controversy about the effect of alcohol, but some studies have shown superiority for EO and CHX with alcohol on cariogenic and periodontopathogenic biofilms, respectively, when compared to alcohol-free version; for CPC, no difference was found. More clinical studies are needed for better understanding the mechanism of action and the differences in performance among the antiplaque agents.(AU)

Effect of Myracrodruon urundeuva and Qualea grandiflora extracts on viability and activity of microcosm biofilm and prevention of enamel demineralization in vitro / Efeito de extratos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade e atividade de biofilme microcosmo e na prevenção da desmineralização do esmalte in vitro

The objective of this study was to evaluate the antimicrobial and anti-caries effects of two plant extracts. The first chapter dealt with a review of the literature whose objective was to discuss the antimicrobial potential of Brazilian natural agents on the biofilm related to dental caries and gingivitis/periodontal disease. The research of the articles was carried out using PubMed. We found a total of 23 papers. Most of the studies were performed using planktonic microorganisms or under clinical trials. Nineteen articles were focused on cariogenic bacteria. From these nineteen articles, eleven were also about periodontopathogenic bacteria. Four studies addressed only periodontopathogenic bacteria. The most tested Brazilian natural agents were green propolis, essential oils of Lippia sidoides and Copaifera sp. Most of the tested agents showed similar results when compared to positive control (essential oils and extracts) or better effect than negative control (green propolis). More studies involving protocols closer to the clinical condition and the use of response variables that allows understanding the mechanism of action of natural agents are necessary before the incorporation of these natural agents into dental products. The second chapter aimed to test the effect of the hydroalcoholic extracts of Myracrodruon urundeuva All. and Qualea grandiflora Mart. leaves on the viability of the microcosm biofilm and on the prevention of enamel demineralization. The microcosm biofilm was produced on bovine enamel, using human saliva pool mixed with McBain saliva (0.2% sucrose) for 14 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva at 100, 10 and 0.1 µg/ml and Q. grandiflora at 100 and 0.1 µg/ml reduced cell viability similarly to the positive control and significantly more than negative control. M. urundeuva at 1000, 100 and 0.1 µg/ml were able to reduce the counting formation unit-CFU counting of lactobacilli sp. and Streptococcus mutans, while Q. grandiflora at 1000 and 1.0 µg/ml significantly reduced the S. mutans CFU counting. On the other hand, the natural extracts did not reduce the production of extracellular polyssacharides, lactic acid and the development of enamel caries lesions. The third chapter aimed to evaluate the effect of hydroalcoholic extracts of M. urundeuva and Q. grandiflora (alone or combined) on the viability of S. mutans biofilm and the prevention of enamel demineralization. S. mutans strain (ATCC 21175) was reactivated in BHI broth. Minimum inhibitory concentration, minimum bactericidal concentration, minimum biofilm inhibitory concentration and minimum biofilm eradication concentration were determined to choose the concentrations to be tested under the biofilm model. S. mutans biofilm (5x105 CFU/ml) was produced on bovine enamel using McBain saliva with 0.2% sucrose for 3 days. The biofilm was treated daily with the extracts for 1 min. M. urundeuva (isolated or combined) at concentrations equal or higher than 0.625 mg/ml was able to reduce the bacteria viability, whereas Q. grandiflora extract alone showed antimicrobial effect at 5 mg/ml only (p<0.05). On the other hand, none of the extracts was able to reduce the development of enamel caries lesions. Despite the tested natural extracts have antimicrobial effect; they are unable to prevent caries in enamel.(AU)

O objetivo foi avaliar os efeitos antimicrobiano e anti-cárie de dois extratos de plantas. O primeiro capítulo se referiu a uma revisão da literatura cujo objetivo foi discutir o potencial antimicrobiano dos agentes naturais brasileiros sobre o biofilme relacionado à cárie dentária e à gengivite/doença periodontal. A pesquisa dos artigos foi realizada usando o PubMed. Foram encontrados 23 trabalhos. A maioria dos estudos foi realizada utilizando microorganismos na fase planctônica ou ensaios clínicos. Dezenove artigos foram focados em bactérias cariogênicas. Dos dezenove artigos, onze também eram sobre bactérias periodontopatogênicas. Quatro estudos abordaram apenas bactérias periodontopatogênicas. Os agentes naturais brasileiros mais testados foram própolis verde, óleos essenciais de Lippia sidoides e Copaifera sp. Os agentes testados apresentaram resultados similares quando comparados ao controle positivo (óleos essenciais e extratos) ou melhor efeito que o controle negativo (própolis verde). Mais estudos próximos da condição clínica e o uso de variáveis de resposta que permitam entender o mecanismo de ação são necessários, para permitir a incorporação desses agentes naturais em produtos odontológicos. O segundo capítulo teve como objetivo testar o efeito dos extratos hidroalcoólicos de Myracrodruon urundeuva All. e Qualea grandiflora Mart. sobre a viabilidade do biofilme microcosmo e na prevenção da desmineralização do esmalte. O biofilme microcosmo foi produzido em esmalte bovino, utilizando pool de saliva humana misturada à saliva de McBain (0,2% de sacarose) durante 14 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva a 100, 10 e 0,1 µg/ml e Q. grandiflora a 100 e 0,1 µg/ml reduziram a viabilidade dos microrganismos de forma semelhante ao controle positivo e significativamente maior do que o controle negativo. M. urundeuva a 1000, 100 e 0,1 µg/ml foi capaz de reduzir a contagem de Unidade formadora de colônia-UFC para Lactobacilos totais e Streptococcus mutans, enquanto a Q. grandiflora a 1000 e 1,0 µg/ml reduziu significativamente a contagem de UFC para S. mutans. Os extratos naturais não conseguiram reduzir a produção de polissacarídeos extracelulares-PEC, ácido lático e o desenvolvimento da lesão cariosa em esmalte. O terceiro capítulo teve como objetivo avaliar o efeito dos extratos hidroalcoólicos de M. urundeuva. e Q. grandiflora (sozinhos ou combinados) sobre a viabilidade do biofilme de S. mutans e na prevenção da desmineralização do esmalte. Cepa de S. mutans (ATCC 21175) foi reativada em caldo BHI. Concentração inibitória mínima, concentração bactericida mínima, concentração inibitória mínima de biofilme e concentração de erradicação mínima de biofilme foram determinadas para escolher as concentrações a serem testadas sob o modelo de biofilme. O biofilme de S. mutans (5x105 CFU/ml) foi produzido em esmalte bovino, utilizando saliva de McBain com 0,2% de sacarose durante 3 dias. O biofilme foi tratado diariamente com os extratos durante 1 min. M. urundeuva (isolada ou combinada) nas concentrações iguais ou superiores a 0,625 mg/ml foi capaz de reduzir a viabilidade das bactérias, enquanto que o extrato da Q. grandflora apresentou efeito antimicrobiano somente a 5 mg/ml (p<0,05). Nenhum dos extratos reduziu o desenvolvimento da lesão da cárie. Apesar dos extratos naturais terem efeito antimicrobiano, são incapazes de prevenir o desenvolvimento da lesão cariosa em esmalte.(AU)

Comparação dos meios de cultura e das técnicas de quantificação de bactérias e fungos em reservatórios e tubulações de água de equipos odontológicos / Comparison of media and techniques for bacteria and fungi quantification in reservoirs and dental unit waterlines

Foram selecionados, aleatoriamente, 12 equipos em 7 clínicas da FOB/USP, dois foram preenchidos com água destilada (Ortodontia e Urgência), um com água de torneira (UBAs), em um o reservatório ficou seco por 30 dias (Odontopediatria) em outro por 60 dias (Pós-graduação), um equipo com a tecnologia B-Safe® (Multidisciplinar) e, em três, a limpeza com o detergente enzimático foi avaliado (Dentística). Amostras de 10 mL de água dos reservatórios e tubulações das canetas de alta rotação foram obtidas e diluições feitas até 10-4, semeadas nos meios de cultura R2A, Peptona Diluída - PD, Plate Count Agar - PCA e Sabouraund Dextrose Agar SDA. Alíquotas de 100 L das amostras foram semeadas nos meios de cultura R2A, PD, PCA e SDA pela técnica de esgotamento, alíquotas de 25 L foram semeadas (R2A, PD, PCA e SDA) pela técnica da gota e alíquotas de 100 L das amostras foram semeadas no meio PCA pela técnica de pour plate. As placas de R2A, PD, PCA foram incubadas por 72 horas a 24°C e as placas de SDA por 4 a 7 dias a 24°C. Foi feita a identificação bacteriana através do kit Bactray I, II ou III e a fúngica através do microcultivo. A média bacteriana obtida foi de 128.151 UFC/mL na Odontopediatria, 1.834.807 UFC/mL na Ortodontia, 60.422 UFC/mL na Pós-Graduação, 615,68 UFC/mL na UBAs, 899,64 UFC/mL na Urgência, 97.632 UFC/mL na Multidisciplinar e 417.619 UFC/mL na Dentística sem limpeza e 135.924 UFC/mL após a limpeza. A média dos fungos foram 205 UFC/mL na Odontopediatria, 702,50 UFC/mL na Ortodontia, 12,50 UFC/mL na Pós-Graduação, 41.475 UFC/mL UBAs, 117.500 UFC/mL Urgência, 4.469 UFC/mL na Multidisciplinar e 64.642 UFC/mL e de 23.627 UFC/mL, antes e após a limpeza na Dentística. A presença de micro-organismos foi detectada nos reservatórios e tubulações de água em todas as 7 clínicas; para quantificar as bactérias, o meio R2A seguido do PD foram melhores que o PCA e, para detectar diferentes espécies o meio PCA foi superior ao R2A e PD; a técnica da gota...

Twelve dental units of 7 clinics of FOB/USP were randomly selected, two were filled with distilled water (Orthodontics and Urgency), one with tap water (UBAs), one reservoir was dry for 30 days (Odontopediatry) and another for 60 days (Postgraduate Clinic), one dental unit was filled with the B - Safe ® technology (Multidisciplinary Clinic). Three dental clinics were evaluated concerning the cleaning procedure with enzymatic detergent. Samples of 10 mL of water reservoirs and high-speed handpieces were collected and made up to 10-4 dilutions, plated on R2A media, Peptone Diluted culture - PD, Plate Count Agar - PCA and Sabouraund Dextrose Agar - SDA. Aliquots of 100 L of the samples were plated on R2A media, PD, PCA and SDA culture technique for spreading. Aliquots of 25 L were seeded (R2A, PD, PCA and SDA) in drops and aliquots of 100 L samples were seeded in PCA by the pour plate technique. R2A plates, PD, PCA were incubated for 72 hours at 24°C and SDA plates for 4 to 7 days at 24°C. Bacterial identification was performed using Bactray I, II or III kit and fungal identification by microculture. Bacterial average was 128.151 CFU/mL in Odontopediatry, 1.834.807 CFU/mL in Orthodontics, 60.422 CFU/mL in Postgraduate Clinic, 615.68 CFU/mL in UBAs, 899.64 CFU/mL in Urgency, 97.632 CFU/mL in Multidisciplinary Clinic and 417.619 CFU/mL in Dentistry without the cleaning procedure and 135.924 CFU/mL after it. The average of fungi was 205 CFU/mL in Odontopediatry, 702.50 CFU/mL in Orthodontics, 12.50 CFU/mL in Postgraduate Clinic, 41.475 CFU/mL in UBAs, 117.500 CFU/mL in Urgency, 4.469 CFU/mL in Multidisciplinary and 64.642 CFU/mL and 23.627 CFU/mL before and after cleaning procedure in Dentistry. The presence of micro - organisms occurred in reservoirs and waterlines in all of the 7 clinics evaluated. To quantify bacteria, R2A and PD medium provided more accurate results than PCA. To detect different species, PCA medium was better than R2A and PD. The drop...