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Master's Dissertation
DOI
10.11606/D.25.1999.tde-03012005-155153
Document
Author
Full name
Cláudio Maníglia Ferreira
Institute/School/College
Knowledge Area
Date of Defense
Published
Bauru, 1999
Supervisor
Committee
Rosa, Odila Pereira da Silva (President)
Moraes, Ivaldo Gomes de
Silva, Salete Moura Bonifacio da
Title in Portuguese
Avaliação 'In Vitro' da Atividade Antimicrobiana de Substâncias Utilizadas em Endodontia sobre Bactérias Anaeróbias.
Keywords in Portuguese
endodontia
microbiologia bucal
Abstract in Portuguese
Avaliou-se “in vitro” a capacidade antimicrobiana de substâncias utilizadas como curativo de demora em Endodontia (soluções de hidróxido de cálcio a 10%, paramonoclorofenol canforado, digluconato de clorexidina a 2% e detergente de mamona a 10%) sobre bactérias anaeróbias (Fusobacterium nucleatum ATCC 25586, Prevotella intermedia ATCC 33563 e Clostridium perfringens ATCC 13124). Os microrganismos foram recebidos liofilizados, reconstituídos, cultivados e estocados a -20oC até o momento de sua utilização, quando então foram reativados em caldo Reinforced Clostridial Medium (RCM), em caldo Brucella suplementado com hemina e vitamina k1 e também em placas de ágar dos mesmos meios acrescidos de 5% de sangue de carneiro. As substâncias testadas foram diluídas nos caldos RCM e Brucella suplementados, em microplacas e tubos. O inóculo foi preparado a partir de repiques do crescimento bacteriano nos dois caldos, buscando-se a concentração de 5 x 105 UFC/ml, avaliada através de leitura em espectrofotômetro. Quantidades iguais do inóculo foram acrescentadas às diluições das drogas, seguindo-se a incubação em anaerobiose. Em todos os testes foram utilizados controles positivos e negativos. Não foi possível a análise visual do crescimento no micrométodo, para determinar a concentração inibitória mínima (CIM); por essa razão, foi determinada a concentração bactericida mínima (CBM) em ágar sangue. Após a incubação dos caldos RCM (48 horas) e Brucella (96 horas), avaliou-se visualmente e através de espectrofotômetro a turvação do meio, para estabelecer a CIM, e semearam-se todos os tubos sem crescimento, para determinação da CBM. A microdiluição não se mostrou confiável para todas as drogas, na comparação com a macrodiluição. Observou-se que o caldo RCM apresenta melhor rendimento que o caldo Brucella. A clorexidina foi a droga que demonstrou melhor eficiência, com as menores CIMs, seguida pelo detergente de mamona, paramonoclorofenol canforado e hidróxido de cálcio. Observou-se, também, que a eficiência de cada droga varia para diferentes bactérias. O Clostridium perfringens exibiu maior resistência que o Fusobacterium nucleatum e a Prevotella intermedia, cujas CBMs foram, em geral, semelhantes para as diferentes drogas.
Title in English
“In vitro” evaluation of the antimicrobial activity of endodontics medicaments against anaerobic bacteria
Abstract in English
The antimicrobial activity of chemicals used as intracanal dressing in Endodontics (calcium hydroxide 10%, camphorated paramonochlorophenol, chlorhexidine digluconate 2% and mamone detergent 10% solutions) against anaerobic bacteria (Fusobacterium nucleatum ATCC 25586, Prevotella intermedia ATCC 33563 and Clostridium perfringens ATCC 13124) was evaluated, using broth microdilution and macrodilution methods. The microorganisms were reference strains received lyophilized, that were reconstituted, cultivated and maintained at -20o C until use, when they were reactivated using Reinforced Clostridial Medium (RCM) and Brucella broth supplemented with hemin and vitamin k1, and also the same agar media with 5% blood sheep. The chemicals were diluted in both broths in microplates and tubes. The inoculum was prepared in the same broths from subcultures made in each of them, that means a concentration of 5 x 105 bacteria/ml, and equal amounts were added to the drugs dilutions, proceeding after the incubation at 37o C in anaerobic jars. It was not possible the visual analysis of bacterial growth by the micromethod, for the minimal inibition concentration (MIC) determination, thence the determination of minimal bactericidal concentration (MBC) by the subculture on blood agar plates. The media turbidity in macromethod was visually and by spectrophotometer evaluated after 48 hours and 96 hours incubation of RCM and Brucella broths, respectively, to determine the CIM and from that, the CBM, in agar media. The analysis of methods showed that the microdilution method is not trustful for all chemicals in comparing to macrodilution method and also, that the RCM broth works better than Brucella broth. There was more efficient activity of chlorhexidine digluconate, followed by mamone detergent, camphorated paramonochlorophenol and calcium hydroxide. The efficiency of each drug varies for different bacteria. The highest resistance was seen with Clostridium perfringens, with Fusobacterium nucleatum and Prevotella intermedia showing similar CBMs for different drugs.
 
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Publishing Date
2005-01-18
 
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