Avaliação da atividade antimicrobiana do extrato etanólico bruto da casca da sucupira branca (Pterodon emarginatus Vogel) - Fabaceae / Evaluation of antimicrobial activity of crude ethanol extract from the bark of "sucupira branca" (Pterodon emarginatus Vogel) - Fabaceae

Pterodon emarginatus Vogel (Fabaceae) é uma árvore do Cerrado conhecida popularmente como "sucupira branca, faveiro, fava de sucupira e sucupira lisa" e utilizada na medicina popular em preparações anti-reumáticas, antiinflamatórias, analgésicas e antiinfecciosas. Esse trabalho teve por objetivo fazer a triagem fitoquímica do pó e avaliar a atividade antimicrobiana do extrato etanólico bruto das cascas da P. emarginatus contra bactérias Gram-positivas, Gram-negativas e o fungo Candida albicans. O extrato etanólico bruto foi obtido a partir das cascas dessecadas e pulverizadas. A concentração inibitória mínima (CIM) do extrato bruto foi determinada utilizando-se o inoculador de Steers. Os testes fitoquímicos detectaram a presença de flavonóides, heterosídeos saponínicos, resinas e traços de esteróides e triterpenóides. As CIM do extrato etanólico foram de 0,18 mg mL-1 para as bactérias Gram-positivas Rhodococcus equi ATCC 25923, Micrococcus luteus ATCC 9341, Micrococcus roseus IPTSP/UFG e para as bactérias Gram-negativas Serratia marcescens ATCC 14756 e Pseudomonas aeruginosa ATCC 9027; de 0,37 mg mL-1 para a Enterobacter cloacae FT 505 LEMC/EPM/UFG e de 0,74 mg mL-1 para as demais bactérias testadas e para o fungo C. albicans. O presente estudo abre perspectivas para o uso da cascas da P. emarginatus como antimicrobiano.

Pterodon emarginatus Vogel (Fabaceae) is a Cerrado tree popularly known as "sucupira branca", "faveiro", "fava de sucupira" and "sucupira lisa" and has been used in folk medicine as antirheumatic, anti-inflammatory, analgesic and anti-infective. The aim of this work was to perform the phytochemical screening of the powder and to evaluate the antimicrobial activity of crude ethanol extract from P. emarginatus barks against Gram-positive bacteria, Gram-negative bacteria and the fungus Candida albicans. The crude ethanol extract was obtained from desiccated and pulverized barks. Its minimum inhibitory concentration (MIC) was determined by using Steers inoculator. The phytochemical tests detected the presence of flavonoids, saponin heterosides, resins and traces of steroids and triterpenoids. Ethanol extract MICs were 0.18 mg mL-1 for the Gram-positive bacteria Rhodococcus equi ATCC 25923, Micrococcus luteus ATCC 9341 and Micrococcus roseus IPTSP/UFG, and for the Gram-negative bacteria Serratia marcescens ATCC 14756 and Pseudomonas aeruginosa ATCC 9027; 0.37 mg mL-1 for Enterobacter cloacae FT 505 LEMC/EPM/UFG; and 0.74 mg mL-1 for the remaining tested bacteria and for the fungus C. albicans. This study opens perspectives for the use of P. emarginatus barks as an antimicrobial drug.

Genetic heterogeneity of Escherichia coli strains isolated from raw milk, Minas Frescal cheese, and food handlers / Heterogeneidade genética de cepas de Escherichia coli isoladas de leite cru, queijo Minas Frescal e manipuladores

From February 2004 to March 2005, 140 samples of food handlers - hands and nostrils - (92), raw milk (24), and minas frescal cheese (24) were analyzed for the presence of Escherichia coli in a dairy processing plant of Goiás State. Forty-seven E. coli strains were obtained and compared by DNA macrorestriction patterns obtained from pulsed-field gel electrophoresis following XbaI restriction in order to investigate the possible sources of cheese contaminations. Based on PFGE genotyping, one strain isolated from food the hands of a handler and five strains isolated from raw milk were identical or closely related to six strains from cheese suggesting, in these cases, the probable source of E. coli contamination in cheeses. No strain isolated from the nostrils was related to those found in cheeses or milk strains. The results showed high diversity among the strains, demonstrating a lack of predominance of an endemic clone in the dairy plant. This paper highlights the usefulness of PFGE as an epidemiological tool for determining the source of E. coli contamination in the food industry.(AU)

Durante um ano, de fevereiro de 2004 a março de 2005, 140 amostras retiradas das mãos e das narinas de manipuladores de alimentos (92), do leite cru (24) e do queijo-de-minas frescal (24) foram analisadas para a presença de Escherichia coli, em um laticínio do Estado de Goiás. As 47 cepas obtidas foram comparadas por macrorrestrição do DNA com enzima XbaI, seguida de eletroforese em gel em campo pulsado (PFGE), a fim de investigar as possíveis fontes de contaminação do queijo. Baseado na genotipagem pelo PFGE, uma cepa obtida do leite cru e cinco cepas obtidas dos manipuladores mostraram similaridade maior que 80 por cento com seis cepas isoladas do queijo, denotando forte correlação genética entre elas e sugerindo, nestes casos, a fonte provável de contaminação do produto final. Nenhuma cepa isolada do nariz foi relacionada às isoladas do queijo ou do leite. Os resultados mostraram grande diversidade entre as cepas, demonstrando ausência de um clone endêmico no laticínio avaliado. Este estudo destaca a utilidade do PFGE como uma ferramenta importante em investigações epidemiológicas e na determinação de possíveis fontes de contaminação por E. coli na indústria de alimentos.(AU)

Genetic heterogeneity of Escherichia coli strains isolated from raw milk, Minas Frescal cheese, and food handlers / Heterogeneidade genética de cepas de Escherichia coli isoladas de leite cru, queijo Minas Frescal e manipuladores

From February 2004 to March 2005, 140 samples of food handlers - hands and nostrils - (92), raw milk (24), and minas frescal cheese (24) were analyzed for the presence of Escherichia coli in a dairy processing plant of Goiás State. Forty-seven E. coli strains were obtained and compared by DNA macrorestriction patterns obtained from pulsed-field gel electrophoresis following XbaI restriction in order to investigate the possible sources of cheese contaminations. Based on PFGE genotyping, one strain isolated from food the hands of a handler and five strains isolated from raw milk were identical or closely related to six strains from cheese suggesting, in these cases, the probable source of E. coli contamination in cheeses. No strain isolated from the nostrils was related to those found in cheeses or milk strains. The results showed high diversity among the strains, demonstrating a lack of predominance of an endemic clone in the dairy plant. This paper highlights the usefulness of PFGE as an epidemiological tool for determining the source of E. coli contamination in the food industry.

Durante um ano, de fevereiro de 2004 a março de 2005, 140 amostras retiradas das mãos e das narinas de manipuladores de alimentos (92), do leite cru (24) e do queijo-de-minas frescal (24) foram analisadas para a presença de Escherichia coli, em um laticínio do Estado de Goiás. As 47 cepas obtidas foram comparadas por macrorrestrição do DNA com enzima XbaI, seguida de eletroforese em gel em campo pulsado (PFGE), a fim de investigar as possíveis fontes de contaminação do queijo. Baseado na genotipagem pelo PFGE, uma cepa obtida do leite cru e cinco cepas obtidas dos manipuladores mostraram similaridade maior que 80 por cento com seis cepas isoladas do queijo, denotando forte correlação genética entre elas e sugerindo, nestes casos, a fonte provável de contaminação do produto final. Nenhuma cepa isolada do nariz foi relacionada às isoladas do queijo ou do leite. Os resultados mostraram grande diversidade entre as cepas, demonstrando ausência de um clone endêmico no laticínio avaliado. Este estudo destaca a utilidade do PFGE como uma ferramenta importante em investigações epidemiológicas e na determinação de possíveis fontes de contaminação por E. coli na indústria de alimentos.

Análise da atividade antimicrobiana das folhas de Davilla elliptica St.-Hil. (Dilleniaceae) / Investigation of antimicrobial activity of Davilla elliptica St.-Hil. leaves (Dilleniaceae)

Davilla elliptica St.-Hil. (Dilleniaceae) é um arbusto,usado popularmente como adstringente, tônico, sedativo, diurético, no tratamento de hemorróidas, hérnia e em aplicações tópicas como anti-séptico na limpeza de ferimentos. O presente estudo teve por objetivo avaliara atividade antimicrobiana dos extratos etanólicos brutos das folhas de D. elliptica coletadas em Alexânia e Goiânia, Goiás, Brasil. Os extratos etanólicos brutos foram obtidos, por maceração a frio, a partir das folhas dessecadas e pulverizadas. A atividade antimicrobiana foi avaliada utilizando 25 cepas de microrganismos (Gram-negativos, Gram-positivos e levedura) através da determinação da Concentração Inibitória Mínima (CIM) utilizando-se o inoculador de Steers. Os extratos obtidos de plantas coletados nas duas localidades inibiram o crescimento de todas as cepas avaliadas. O extrato da amostra coletada em Alexânia apresentou atividade antimicrobiana com CIM variando de 0,371 mg/mL a 0,743 mg/mL. O extrato da amostra coletada em Goiânia apresentou CIM variando de < 0,023 mg/mL a 11,9 mg/mL. Os extratos etanólicos brutos das folhas de D. elliptica foram mais ativos contra as bactérias Gram-positivas do que para as bactérias Gram-negativas.

Davilla elliptica St.-Hil.(Dilleniaceae) is a shrub, whose leaves are traditionally used in Brazilian popular medicine as an astringent, tonic, sedative and diuretic and to treat hemorrhoids and hernia and it is applied a topical antiseptic to clean wounds. The aim in this study was to assess the antimicrobial activity of crude ethanol extracts of D. elliptica leaves collected in the towns of Alexânia and Goiânia, state of Goiás, Brazil. The crude extracts were obtained by cold maceration, of the dried and pulverized leaves in ethanol. Antimicrobial activities against 25 strains of microorganisms (Gram-negative bacteria, Gram-positive bacteria and a yeast) were tested determining by the Minimal Inhibitory Concentration (MIC) in agar plates inoculated with using a Steers replicator. Extracts of from inhibited the growth of all strains evaluated. The extract of the sample collected in Alexânia showed antimicrobial activity with MICs ranging from 0.371 mg/mL to 0.743 mg/mL, while that collected in Goiânia showed MICs ranging from <0.023 mg/mL to 11.9 mg/mL. The D. elliptica crude leaf ethanol extracts evaluated were more active against Gram-positive than Gram-negative bacteria.

Avaliação da atividade antimicrobiana das partes aéreas (folhas e caules) e raízes de Richardia brasiliensis Gomez (Rubiaceae) / Assessment of antimicrobial activity of aerial parts (leaves and stems) and roots of Richardia brasiliensis Gomez (Rubiaceae)

Richardia brasiliensis é uma planta utilizada popularmente como expectorante, emética, diaforética, vermífuga e para o tratamento de hemorróidas. O presente estudo objetivou realizar a prospecção fitoquímica e avaliar a ação antimicrobiana do extrato bruto das partes aéreas e raízes de R. brasiliensis. O extrato etanólico bruto foi obtido a partir do material botânico dessecado e pulverizado. O pó das partes aéreas e raízes foram submetidos à triagem fitoquímica. A atividade antimicrobiana foi realizada contra bactérias Gram-positivas esporuladas e não esporuladas, Gram negativas e a levedura Candida albicans através da determinação da concentração inibitória mínima por diluição em ágar empregando-se o inoculador de Steers. A triagem fitoquímica evidenciou a presença de esteróides, triterpenóides, cumarinas, resinas, alcaloides e flavonóides. A CIM do extrato das partes aéreas variou de 0,37 a 0,74 mg/mL e das raízes de 0,74 a 11,9 mg/mL. Concluiu-se que tanto o extrato etanólico bruto das partes aéreas quanto das raízes apresentaram atividade antimicrobiana.

Richardia brasiliensis is a neotropical perennial her bused in Brazilian popular medicine as an expectorant, emetic, diaphoretic, vermifuge and to treat hemorrhoids. This paper reports the phytochemical screening and the antimicrobial activity of crude ethanol extracts of the aerial parts and roots of this plant. The crude ethanol extracts were prepared from dried, pulverized botanical material. The aerial parts and roots powder was screened for classes of phytochemicals. The antimicrobial activity was tested against spore-forming and non-sporing Gram-positive bacteria, Gramnegative bacteria and Candida albicans by the agardilution method, inoculating the series of plates at multiple points with a Steers replicator to determine the minimum inhibitory concentration (MIC) of each organism. The phytochemical screening showed the presence of steroids, triterpenes, coumarins, resins, alkaloids and flavonoids. The MIC of the extract of aerial parts varied from 0.37 to 0.74 mg/mL and that of the roots from 0.74 to 11.9 mg/mL. It was concluded that the crude ethanolic extracts of both the aerial parts and the roots showed antimicrobial activity.

Evaluation of antimicrobial activity of the crude ethanol extract of Eugenia uniflora L. leaves

Eugenia unifl ora L. is a tree whose leaves are used in popular medicine as an antihypertensive, antimicrobial drug, in the treatment of bronchitis, influenza and as an antipyretic. This paper reports the antimicrobial activity of a crude ethanol extract of E. unifl ora L. leaves. The crude extract was prepared from material collected in Goiânia, Goiás (Brazil), dried, pulverized and subjected to phytochemical screening. The antimicrobial activity was tested against spore-forming and non-sporing Gram-positive and Gram-negative bacteria as well as Candida albicans, using the well-diffusion test and the agar dilution method to determine the minimum inhibitory concentration (MIC). The phytochemical screening showed the presence of tannins, steroids, triterpenes, heterosides, anthraquinones, flavonoids and saponins. Antimicrobial activity testing showed that the crude E.uniflora L. leaf extract inhibited all the Gram-positive bacteria (MICs from 0.273 to 8.75 mg/mL), among which the spore-formers exhibited MICs from 1.094 to 2.187 mg/mL. The MIC for most Gram-negative bacteria varied from 4.375 to 17.5 mg/mL. C. albicans NTC 2010 (MIC of 0.547 mg/mL) inhibition was also noted. The antimicrobial activity found in this in vitro study of E. uniflora L. may justify its popular use as a medicine.

Phenotypic aspects of oral strains of Candida albicans in children with Down's syndrome.

The aim of this article is to characterize the biological aspects of oral strains of C. albicans in children with Down's syndrome. These yeasts were analyzed as to their macromorphological and enzymatic aspects and were tested as to their in vitro susceptibility to antifungal drugs using broth microdilution to determine the minimum inhibitory concentration (MIC). The morphotyping revealed that all oral C. albicans isolates from children with Down's syndrome promoted the formation of fringes regardless of size, while the control group presented smaller fringes. All oral C. albicans strains produced proteinase, but those with phospholipolytic activity showed greater enzyme capacity in the test group. In vitro susceptibility showed that all oral C. albicans isolates were sensitive to the drugs used.

Phenotypic aspects of oral strains of Candida albicans in children with down's syndrome / Aspectos fenotípicos de cepas de Candida albicans orais em crianças com síndrome de down

The aim of this article is to characterize the biological aspects of oral strains of C. albicans in children with Down's syndrome. These yeasts were analyzed as to their macromorphological and enzymatic aspects and were tested as to their in vitro susceptibility to antifungal drugs using broth microdilution to determine the minimum inhibitory concentration (MIC). The morphotyping revealed that all oral C. albicans isolates from children with Down's syndrome promoted the formation of fringes regardless of size, while the control group presented smaller fringes. All oral C. albicans strains produced proteinase, but those with phospholipolytic activity showed greater enzyme capacity in the test group. In vitro susceptibility showed that all oral C. albicans isolates were sensitive to the drugs used.

O objetivo deste artigo foi caracterizar os aspectos biológicos de cepas de C. albicans orais em crianças com síndrome de Down. Estas leveduras foram analisadas quanto aos seus aspectos macromorfológicos e enzimáticos e foram testadas quanto a sua suscetibilidade in vitro a drogas antifúngicas, usando a microdiluição em caldo para a determinação da concentração inibitória mínima (CIM). A morfotipagem revelou que todos os isolados de C. albicans orais de crianças com síndrome de Down induziram à formação de franjas independente do tamanho, enquanto o grupo controle teve franjas menores. Todas as cepas de C. albicans orais produziram proteinase, mas aquelas com atividade fosfolipidolítica mostraram maior capacidade enzimática no grupo teste. A suscetibilidade in vitro mostrou que todos os isolados de C. albicans orais foram sensíveis a drogas empregadas.

Intranasal immunization with the cholera toxin B subunit-pneumococcal surface antigen A fusion protein induces protection against colonization with Streptococcus pneumoniae and has negligible impact on the nasopharyngeal and oral microbiota of mice.

One of the candidate proteins for a mucosal vaccine antigen against Streptococcus pneumoniae is PsaA (pneumococcal surface antigen A). Vaccines targeting mucosal immunity may raise concerns as to possible alterations in the normal microbiota, especially in the case of PsaA, which was shown to have homologs with elevated sequence identity in other viridans group streptococci. In this work, we demonstrate that intranasal immunization with a cholera toxin B subunit-PsaA fusion protein is able to protect mice against colonization with S. pneumoniae but does not significantly alter the natural oral or nasopharyngeal microbiota of mice.

Intranasal immunization with the cholera toxin B subunit-pneumococcal surface antigen A fusion protein induces protection against colonization with Streptococcus pneumoniae and has negligible impact on the nasopharyngeal and oral microbiota of mice

One of the candidate proteins for a mucosal vaccine antigen against Streptococcus pneumoniae is PsaA (pneumococcal surface antigen A). Vaccines targeting mucosal immunity may raise concerns as to possible alterations in the normal microbiota, especially in the case of PsaA, which was shown to have homologs with elevated sequence identify in other viridans group streptococci. In this work, we demonstrate that intranasal immunization with a cholera toxin B subunit-PsaA fusion protein is able to protect mice against colonization with S. pneumoniae but does not significantly alter the natural oral or nasopharyngeal microbiota of mice.

Serotypes of carriage and invasive isolates of Streptococcus pneumoniae in Brazilian children in the era of pneumococcal vaccines.

Nasopharyngeal carriage of Streptococcus pneumoniae is a key factor in the development of invasive disease and the spread of resistant strains within the community. A single nasopharyngeal swab was obtained from 648 unvaccinated children aged <5 years, either healthy or with acute respiratory tract infection or meningitis, during the winters of 2000 and 2001. The overall pneumococcal carriage rate was 35.8% (95% CI 32.1-39.6). The pneumococcal serotypes found most frequently in the nasopharynx were 14, 6B, 6A, 19F, 10A, 23F and 18C, which included five of the seven serotypes in the currently licensed seven-valent conjugate vaccine (PCV7); serotypes 4 and 9V were less common. Serotypes 1 and 5 were isolated rarely from the nasopharynx. A comparison of 222 nasopharyngeal isolates with 125 invasive isolates, matched for age and time to the carrier isolates, showed a similar prevalence of penicillin non-susceptible pneumococci (PNSp) (19.8% and 19.2%, respectively). PNSp serotypes were similar (6B, 14, 19F, 19 A, 23B and 23F) for carriage and invasive disease isolates. The coverage of PCV7 for carriage isolates (52.2%) and invasive isolates (62.4%) did not differ significantly (p 0.06); similarly, there was no significant difference in PCV7 coverage for carriage isolates (34.5%) and invasive isolates (28.2%) of PNSp. These data suggest that PCV7 has the potential to reduce pneumococcal carriage and the number of carriers of PNSp belonging to vaccine serotypes.

Cockroaches as carriers of fungi of medical importance.

The fungal pathogenic flora of the external surface of 103 cockroaches (Periplaneta americana) collected from the intensive care unit of a hospital were investigated. In this study, a high percentage of test cockroaches (93.2%) were found to carry fungi of medical importance. The main fungi isolated were species of Candida, Aspergillus and Penicillium. Information about the carriage of pathogenic fungi by cockroaches in hospital environment is scanty. The results suggest that cockroaches can play a role in dissemination of fungi, which they can carry on their external surface.

Prevalence of bacteria in the nipple discharge of patients with duct ectasia.

The aim of this study was to identify the aerobic and the anaerobic microorganisms which can be related to duct ectasia. The patients were divided into two groups. Group 1 comprised 100 patients with coloured nipple discharge (duct ectasia group), and Group 2 (the control group) was composed of 50 patients without nipple discharge. The culture media used were BHI-PRAS, blood agar, mannitol agar and MacConkey agar. There was a high frequency of bacterial growth in the two groups: 85% in Group 1 and 88% in Group 2. The most prevalent bacteria were Staphylococcus aureus and Staphylococcus epidermidis. There was a statistically significant higher rate of smokers in the duct ectasia group compared with the control group, 25 (25%) patients vs. 5 (10%), respectively (p = 0.03). These findings allow us to put forth the hypothesis that the genesis of duct ectasia may be a non-infectious inflammatory process.

Efficacy of sodium hypochlorite and coconut soap used as disinfecting agents in the reduction of denture stomatitis, Streptococcus mutans and Candida albicans.

This study evaluated the reduction of denture stomatitis and the antimicrobial activity of 0.05% sodium hypochlorite opposed to Candida albicans and Streptococcus mutans (SGM) when associated with brushing complete dentures with coconut soap. The mucosal characteristics were evaluated according to Newton's classification at baseline, after cleansing the dentures with coconut soap for 15 days in group 1 (nine patients). In the other group (19 patients) the analysis were made before and after cleansing the dentures with coconut soap and with disinfection in a soak solution of 0.05% sodium hypochlorite for 10 min during 15 days. Microbiological tests were used to isolate C. albicans and SGM. Mann-Whitney and Wilcoxon tests were used to compare the mucosal characteristics and Fisher test and McNemar test to compare C. albicans and SGM levels. Statistical analysis at the 95% confidence level (P < 0.05) showed that: (i) the association of coconut soap and 0.05% sodium hypochlorite significantly reduced clinical signs of denture stomatitis, (ii) C. albicans did not reduce in counts, (iii) SGM were reduced but not significantly and (iv) the association of coconut soap and 0.5% sodium hypochlorite was effective in controlling denture biofilm.

Prevalence of mutans streptococci in 93 members from six Brazilian families.

Several studies report that mutans streptococci (MS) are closely associated with caries in humans and that there is a correlation between the number of carious lesions and the levels of MS in the saliva of children and adults. The presence of MS in the saliva of 93 members of six Brazilian families with at least 3 generations was investigated. Samples of whole unstimulated saliva were collected and diluted. Aliquots of 50 microliters of each suspension were dropped onto SB20 agar and incubated in a candle jar at 37 degrees C for 72 h. Colonies resembling MS were counted, collected, seeded in thioglycoilate medium and subjected to biochemical typing. Mutans streptococci were isolated from 80 subjects (86.0%) and the counts ranged from 3.0 x 10(2) (log 2.477) to 1.6 x 10(8) (log 8.204) CFU/ml of saliva. All of the 73 adults were colonized by MS, but the bacteria were detected in only 7 (35.0%) of the 20 children evaluated. Streptococcus mutans occurred in 78 subjects (97.5%), and 51 (63.7%) were monocolonized. S. sobrinus occurred in 29 individuals (36.3%) and 2 (2.5%) were monocolonized. Twenty-seven (33.8%) subjects were multicolonized with S. mutans and S. sobrinus. This study showed a high prevalence (86.0%) of mutans streptococci in the saliva of members of the studied families, which suggests the risk of intrafamilial transmission.

Antimicrobial evaluation of calcium hydroxide in infected dentinal tubules.

The objective of this study was to evaluate the antimicrobial activity of calcium hydroxide in infected dentinal tubules. Four microorganisms, strains of ATCC (Streptococcus faecalis (ATCC-29212), Staphylococcus aureus (ATCC-6538), Bacillus subtilis (ATCC-6633), and Pseudomonas aeruginosa (ATCC-27853)) and one mixture of these were used. These strains were inoculated in brain heart infusion (BHI) and incubated at 37 degrees C for 24 h. Sixty-three human maxillary central incisors were prepared and sterilized by autoclaving. Five groups of 12 teeth each were contaminated for 28 days using new 24-h cultures every 72 h, prepared and adjusted to tube 2 of the MacFarland scale (6 x 10(8) cells/ml). Root canals were then irrigated with 5 ml of saline, dried, and completely filled with calcium hydroxide paste. At intervals of 0, 48, and 72 h, and 7 days, dressings were removed and teeth were immersed in 5 ml of BHI and incubated at 37 degrees C for 48 h to observe the growth and multiplication of the microorganisms. Three uninoculated teeth were maintained in a humid environment as an aseptic control. These teeth were immersed in BHI and maintained at 37 degrees C for 7 days to determine microbial growth. Bacterial growth was shown by turbidity of the culture medium and confirmed by seeding these broths on BHI agar at 37 degrees C for 24 h. The positive BHI tubes were selected, and inoculum was spread on the surface of BHI agar, followed by the same incubation conditions. Gram stain was conducted from BHI growth and from colonies growing on solid medium. Calcium hydroxide in infected dentinal tubules showed no antimicrobial effect on S. faecalis, S. aureus, B. subtilis, P. aeruginosa, or on the bacterial mixture used throughout the experiment.

In vitro determination of direct antimicrobial effect of calcium hydroxide.

The objective of this study was to determine in vitro the time required for calcium hydroxide in direct contact with microorganisms to express its antimicrobial effect. The microorganisms used were: Micrococcus luteus (ATCC-9341), Staphylococcus aureus (ATCC-6538), Fusobacterium nucleatum (ATCC-25586), Pseudomonas aeruginosa (ATCC-27853), Escherichia coli, and Streptococcus sp. The strains were cultivated in Brain Heart Infusion (BHI), with the exception of F. nucleatum (BHI-PRAS). Pure and mixed suspensions of the microorganisms were prepared. Paper cones immersed in these substances were covered with calcium hydroxide paste, and after 0, 1, 2, 6, 12, 24, 48, and 72 h and 7 days they were transferred to an appropriate medium to observe the growth and multiplication of the microorganisms. Incubation was conducted at 37 degrees C for 48 h, according to the requirements of oxygen of each microorganism. The antimicrobial effect of calcium hydroxide was shown to occur after 12 h on M. luteus and F. nucleatum, 24 h on Streptococcus sp, 48 h on E. coli, and 72 h on S. aureus and P. aeruginosa. Mixture II (M. luteus + Streptococcus sp + S. aureus) was sensitive to calcium hydroxide antimicrobial potential after 48 h, whereas mixture I (M. luteus + E. coli + P. aeruginosa), mixture III (E. coli + P. aeruginosa), and mixture IV (S. aureus + P. aeruginosa) were inactivated after 72 h of exposure.