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Photodynamic therapy in spores of Bacillus atrophaeus and Bacillus subtilis: studies with laser, led, methylene blue, Rose Bengal and malachite green

Grant number: 10/14434-4
Support type:Scholarships in Brazil - Master
Effective date (Start): March 01, 2011
Effective date (End): July 31, 2012
Field of knowledge:Biological Sciences - Microbiology
Principal Investigator:Antonio Olavo Cardoso Jorge
Grantee:Michelle Peneluppi Silva
Home Institution: Faculdade de Odontologia (FOSJC). Universidade Estadual Paulista (UNESP). Campus de São José dos Campos. São José dos Campos , SP, Brazil


This study will be to compare and evaluate the action exerted by the laser (L) and the light emitting diode (LED) in spores of Bacillus subtilis and Bacillus atrophaeus in photodynamic therapy, the use of photosensitizers methylene blue (MB), rose bengal (RB) and malachite green (VM). Be used a standard strain of Bacillus atrophaeus (ATCC 9372) and Bacillus subtilis (ATCC 19659). The colonies of microorganisms are cultured for 7 days on Nutrient Agar added to 0.003% manganese and analyzed for the formation of spores (Wirtz-Conklin staining). The isolates will be suspended in sterile distilled water and centrifuged for 10 minutes at 653 Xg. The supernatants are discarded and deposits resuspended in 3 ml of sterile distilled water. The suspension will heat shock at 70°C for 30 minutes, and the presence of spores is checked. The suspensions will be standardized to 10^6 cells / mL in a spectrophotometer. Will be used photosensitizers methylene blue (37.5 ¼M), rose bengal (12.5 ¼M) and malachite green (300 ¼M). As light sources are used laser gallium arsenate (visible red) and light emitting diode (LED). In 96-well plate is added 0.1 ml of Bacillus subtilis or atrophaeus and 0.1 ml of the photosensitizer or NaCl 0.9%. The plate with the sample is stirred for 5, 10 and 30 minutes on an orbital shaker and irradiated. Serial dilutions are obtained from each sample. Aliquots of 0.1 mL of the dilutions are plated on agar plate with brain heart infusion and incubated at 37ºC for 48 hours. The Colony Forming Units per milliliter (CFU / mL) will be counted and log-transformed. The results will be statistically analyzed by analysis of variance (ANOVA) and Tukey test, with significance level of 5%.

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
SILVA, MICHELLE PENELUPPI; CHIBEBE JUNIOR, JOSE; JORJAO, ADELINE LACERDA; DA SILVA MACHADO, ANA KARINA; DE OLIVEIRA, LUCIANE DIAS; JUNQUEIRA, JULIANA CAMPOS; CARDOSO JORGE, ANTONIO OLAVO. Influence of artificial saliva in biofilm formation of Candida albicans in vitro. Brazilian Oral Research, v. 26, n. 1, p. 24-28, JAN-FEB 2012. Web of Science Citations: 8.
Academic Publications
(References retrieved automatically from State of São Paulo Research Institutions)
SILVA, Michelle Peneluppi. Terapia fotodinâmica em esporos de bacillus atrophaeus e bacillus subtilis : estudos com LASER, LED, azul de metileno, rosa bengala e verde malaquita. 2012. 104 f. Master's Dissertation - Universidade Estadual Paulista "Júlio de Mesquita Filho" Faculdade de Odontologia (Campus de São José dos Campos).

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