Staphylococcus aureus growth delay after exposure to low fluencies of blue light (470 nm) / Atraso no crescimento de Staphylococcus aureus após exposição à baixas fluências de luz azul (470 nm)

Antibiotic resistance is one of the greatest challenges to treat bacterial infections worldwide, leading to increase in medical expenses, prolonged hospital stay and increased mortality. The use of blue light has been suggested as an innovative alternative to overcome this problem. In this study we analyzed the antibacterial effect of blue light using low emission parameters on Staphylococcus aureus cultures. In vitro bacterial cultures were used in two experimental approaches. The first approach included single or fractionated blue light application provided by LED emitters (470 nm), with the following fluencies: 16.29, 27.16 and 54.32 J/cm2. For the second approach a power LED (470 nm) was used to deliver 54.32 J/cm2 fractionated in 3 applications. Our results demonstrated that bacterial cultures exposed to fractionated blue light radiation exhibited significantly smaller sizes colonies than the control group after 24 h incubation, however the affected bacteria were able to adapt and continue to proliferate after prolonged incubation time. We could conclude that the hypothetical clinical use of low fluencies of blue light as an antibacterial treatment is risky, since its action is not definitive and proves to be ineffective at least for the strain used in this study.

A resistência a antibióticos é um dos maiores desafios para o tratamento de infecções bacterianas em todo o mundo, levando ao aumento de despesas médicas, prolongamento da internação hospitalar e aumento da mortalidade. O uso da luz azul tem sido sugerido como uma alternativa inovadora para superar esse problema. Neste estudo, analisamos o efeito antibacteriano da luz azul usando parâmetros de baixa emissão em culturas de Staphylococcus aureus. Culturas bacterianas foram usadas em duas abordagens experimentais in vitro. A primeira abordagem incluiu o uso da aplicação única ou fracionada de luz azul fornecida por emissores de LED (470 nm), com as seguintes fluências: 16,29, 27,16 e 54,32 J/cm2. Para a segunda abordagem, um LED de potência (470 nm) foi usado para fornecer 54,32 J/cm2 fracionado em 3 aplicações. Nossos resultados demonstraram que as culturas bacterianas expostas à radiação de luz azul fracionada exibiram colônias de tamanhos significativamente menores do que o grupo controle após 24 h de incubação, no entanto, as bactérias afetadas foram capazes de se adaptar e continuar a proliferar após um tempo prolongado de incubação. Podemos concluir que o uso clínico hipotético de baixas fluências de luz azul como tratamento antibacteriano é arriscado, pois sua ação não é definitiva e mostra-se ineficaz, pelo menos para a cepa utilizada neste estudo.

The effects of photobiomodulation therapy on Staphylococcus aureus infected surgical wounds in diabetic rats. A microbiological, histopathological, and biomechanical study

ABSTRACT PURPOSE: To evaluate the effects of photobiomodulation therapy (PBMT) at 685 nm on diabetic wound healing in rats suffering from bacterial infection induced by Staphylococcus aureus (S. aureus). METHODS: Thirty streptozotocin-induced diabetic rats were allocated into two groups: control and PBMT. A 4-cm full-thickness linear-incision was made on the dorsal midline and was contaminated with S. aureus. The wounds in the PBMT group were irradiated daily for 5 consecutive days, starting 3 days after the induction and always in the mornings. RESULTS: The result revealed that PBMT resulted in a significant decrease in S. aureus CFU in the PBMT group in comparison to the control group (P<0.05). The length of wounds, in the 2nd and 3rd weeks, in the PBMT group were significantly shorter compared to the control group (P<0.05). PBMT caused a significant increase in the histological parameters in comparison to the control group (P<0.05). Moreover, PBMT significantly increased the breaking strength of the surgical scars produced in the skin of the PBMT group when compared to the control group (P<0.05). CONCLUSION: Photobiomodulation therapy may be useful in the management of wound infection through a significant bacterial growth inhibition and an acceleration of wound healing process.

The effects of low level laser therapy on Staphylococcus aureus infected third-degree burns in diabetic rats

PURPOSE: To characterize the effects of low-level laser (LLL) on third-degree burn wounds which were infected with Staphylococcus aureus (S. aureus) in diabetic rats. METHODS: Thirty streptozotocin-induced diabetic rats were divided into two groups: the control and the LLL groups. Third-degree burns were induced using a heated metal rod, and then, were contaminated with S. aureus. The wounds in the LLL group were irradiated with a LLL (685nm) daily for five consecutive days, starting three days after the induction. The wound area was measured at 3, 5, 8, 14 and 21 days after burning. At the end of trial, the skin samples were harvested. RESULTS: Reduction in wound areas in the LLL and control groups were significantly different only on the 21st day (p<0.05). The mean bacterial numbers in the LLL group were significantly lower (p<0.05) than those in the control group. The number of macrophages, new blood vessels, fibroblast, and elevated collagen deposition in the LLL group significantly increased compared to the control group (p<0.05). The mean breaking strength of scars in the control group was significantly lower (p<0.05) than that of the LLL group. CONCLUSION: The low-level laser improved the healing of S. aureus third-degree burn infections in diabetic rats.

Effects of low intensity laser in in vitro bacterial culture and in vivo infected wounds / Efeitos do laser de baixa intensidade em cultura bacteriana in vitro e ferida infectada in vivo

OBJECTIVE: to compare the effects of low intensity laser therapy on in vitro bacterial growth and in vivo in infected wounds, and to analyze the effectiveness of the AsGa Laser technology in in vivo wound infections. METHODS: in vitro: Staphylococcus aureus were incubated on blood agar plates, half of them being irradiated with 904 nm wavelength laser and dose of 3J/cm2 daily for seven days. In vivo: 32 male Wistar rats were divided into control group (uninfected) and Experimental Group (Infected). Half of the animals had their wounds irradiated. RESULTS: in vitro: there was no statistically significant variation between the experimental groups as for the source plates and the derived ones (p>0.05). In vivo: there was a significant increase in the deposition of type I and III collagen in the wounds of the infected and irradiated animals when assessed on the fourth day of the experiment (p=0.034). CONCLUSION: low-intensity Laser Therapy applied with a wavelength of 904nm and dose 3J/cm2 did not alter the in vitro growth of S. aureus in experimental groups; in vivo, however, it showed significant increase in the deposition of type I and III collagen in the wound of infected and irradiated animals on the fourth day of the experiment. .

OBJETIVO: comparar os efeitos da terapia a laser de baixa intensidade no crescimento bacteriano in vitro e em feridas infectadas in vivo, e analisar a efetividade da tecnologia Laser AsGa, em feridas infectadas in vivo. MÉTODOS: in vitro: cepas de Staphylococcus aureus foram incubadas em placas de agar-sangue e irradiadas com laser de 904nm de comprimento de onda e dose de 3J/cm2, diariamente durante sete dias. In vivo: 32 ratos machos Wistar foram distribuídos em Grupo Controle (Não Infectado) e Grupo Experimental (Infectados). Metade dos ratos tiveram suas feridas irradiadas e a outra metade não irradiada, como realizado no estudo in vitro. RESULTADOS: in vitro: não houve variação estatística significativa entre os grupos experimentais, considerando as placas matrizes e derivadas (p>0,05). In vivo: houve aumento significativo na deposição de colágeno tipo I e III na cicatriz do grupo dos animais infectados e irradiados, quando avaliados no quarto dia de experimento (p=0,034). CONCLUSÃO: a Low-Intensity Laser Therapy aplicada com comprimento de onda de 904nm e dose de 3J/cm2, in vitro: não alterou o crescimento de S. aureus nos grupos experimentais. In vivo: mostrou aumento significativo na deposição de colágeno tipo I e III na cicatriz no grupo dos animais infectados e irradiados no quarto dia de experimento. .

Antimicrobial susceptibility of photodynamic therapy (UVA/riboflavin) against Staphylococcus aureus / Suscetibilidade antimicrobiana da terapia fotodinâmica (UVA/riboflavina) contra Staphylococcus aureus

PURPOSE: To assess S. aureus in vitro viability after the exposure to ultraviolet light A (UVA) and riboflavin (B2). METHODS: Samples of S. aureus in 96 well plates (in triplicate) were exposed to riboflavin (B2) and ultraviolet light A (365 nm wavelength) at a power density of 3 mW/cm², 8 mm spot diameter, for 30 minutes. Control groups were prepared as well in triplicate: blank control, ultraviolet light A only, riboflavin only and dead bacteria Control. The bacterial viability was measured using fluorescent microscopy. In order to investigate the occurrence of "viable but non-culturable" microorganisms after treatment, the cell viability was also investigated by plate culture procedure onto a broth medium. Statistical analysis was performed using the triplicate values from each experimental condition. RESULTS: No difference was observed among the treatment group and the control samples (p=1). CONCLUSION: The combination of riboflavin 0.1% and ultraviolet light A at 365 nm did not exhibit antimicrobial activity against oxacillin susceptible S. aureus.

OBJETIVO: Avaliar a viabilidade celular de S. aureus in vitro após a exposição de riboflavina (B2) e luz ultravioleta A (UVA). MÉTODOS: Amostras de S. aureus colocadas em uma placa de 96 poços (em triplicata) foram expostas a riboflavina 0,1% (B2) e luz ultravioleta (comprimento de onda de 365 nm) poder de 3 mW/cm², 8 mm de diâmetro, por 30 minutos. Grupos controles foram também preparados em triplicata: controle branco, somente luz ultravioleta A, somente riboflavina e controle morto. A viabilidade bacteriana foi analisada usando microscópio de fluorescência. Para investigar a ocorrência de micro-organismos "viáveis porem não cultiváveis" a viabilidade celular foi avaliada utilizando-se placas de meio de cultivo bacteriano. Analise estatística foi realizada utilizando-se os valores obtidos em triplicata de cada grupo experimental. RESULTADOS: Nenhuma diferença foi observada entre o grupo tratamento e os grupos controle (p=1). CONCLUSÃO: A combinação riboflavina 0,1% e luz ultravioleta 365 nm de comprimento de onda não demonstrou atividade antimicrobiana contra S. aureus oxacilina sensível.

[Evaluation of ultrasound waves effect on antibiotic resistance Pseudomonas aeruginosa and Staphylococcus aureus isolated from hospital and their comparison with standard species]

Pseudomonas aeruginosa and Staphylococcus aureus are important pathogens that produce wide spread infections. Purpose of this study was to evaluate the antimicrobial effect of ultrasonic irradiation [US] alone and in combination with antibiotic on antibiotic resistance Pseudomonas aeruginosa and Staphylococcus aureus. In this study ultrasonic irradiation [US] in a laboratory-scale batch sonoreactor with low frequency [42 kHz] plate type transducer at 170W of acoustic power was used. The Water samples, were taken from different wards of the 3 teaching hospitals which were affiliated to the Kurdistan University of Medical Sciences to isolate Pseudomonas aeruginosa and Staphylococcus aureus and also to determine their antimicrobial susceptibility pattern. Our results showed that Pseudomonas aeruginosa and Staphylococcus aureus were affected by the ultrasound and the bactericidal effect increased with time. It was found that P. aeruginosa was more susceptible to the ultrasonic treatment than S. aureus. The combination of US with an antibiotic [amoxicillin] enhanced killing of both bacteria over the use of US alone. There were no differences in resistance to ultrasound between isolated strains and standard strains from Persian type culture collection

Evaluation of ultraviolet radiation to control microorganisms adhering to low-density polyethylene films

Efficiency of ultraviolet (UV) radiation in reducing the cell number of Staphylococcus aureus ATCC 25923 and Escherichia coli K-12 adhered to low-density polyethylene (LDPE) films was evaluated. The microorganisms were let to adhere to the surface of LPDE bags for 12h at 18liC, and then submitted to UV radiation at an intensity 196 icW.cm-2, 254nm, for 2 seconds. Staphylococcus aureus was less resistant to UV radiation than E. coli, and the efficiency increased with the increase of the concentration of microbial suspension. After 1500 hours of use the UV radiation intensity of the lamp was reduced from 288 to 78 icW.cm-2, and the higher decrease occurred in the first 100 hours of use. Also, the efficiency of the UV radiation decreased after 1500 hours of use. The number of mesophilic aerobes on the surface of LDPE films was reduced by 90 percent after irradiation with 137 icW.cm-2 for 2 seconds. Atomic force microscopy revealed cracks and crevices and protuberances on the LDPE surface, a topography that can protect the cells from UV radiation, reducing the efficiency of the process. The results showed that UV radiation can be a useful technique for reducing the microbiota adhered to LDPE films.