Biofilm formation by Staphylococcus aureus and Salmonella spp. under mono and dual-species conditions and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite

Abstract The aim of this study was evaluated the biofilm formation by Staphylococcus aureus 4E and Salmonella spp. under mono and dual-species biofilms, onto stainless steel 316 (SS) and polypropylene B (PP), and their sensitivity to cetrimonium bromide, peracetic acid and sodium hypochlorite. The biofilms were developed by immersion of the surfaces in TSB by 10 d at 37 °C. The results showed that in monospecies biofilms the type of surface not affected the cellular density (p > 0.05). However, in dual-species biofilms on PP the adhesion of Salmonella spp. was favored, 7.61 ± 0.13 Log10 CFU/cm2, compared with monospecies biofilms onto the same surface, 5.91 ± 0.44 Log10 CFU/cm2 (p < 0.05). The mono and dual-species biofilms were subjected to disinfection treatments; and the most effective disinfectant was peracetic acid (3500 ppm), reducing by more than 5 Log10 CFU/cm2, while the least effective was cetrimonium bromide. In addition, S. aureus 4E and Salmonella spp. were more resistant to the disinfectants in mono than in dual-species biofilms (p < 0.05). Therefore, the interspecies interactions between S. aureus 4E and Salmonella spp. had a negative effect on the antimicrobial resistance of each microorganism, compared with the monospecies biofilms.

Effect of final irrigation protocols on microhardness reduction and erosion of root canal dentin

Abstract This study aimed to evaluate the effect of final irrigation protocols on microhardness reduction and erosion of root canal dentin. Sixty root canals from mandibular incisors were instrumented and randomly divided into six groups (n = 10) according to the irrigant used: QMiX, 17% EDTA, 10% citric acid (CA), 1% peracetic acid (PA), 2.5% NaOCl (solution control), and distilled water (negative control). The chelating solutions were used to irrigate the canal followed by 2.5% NaOCl as a final flush. After the irrigation protocols, all specimens were rinsed with 10 mL of distilled water to remove any residue of the chemical solutions. Before and after the final irrigation protocols, dentin microhardness was measured with a Knoop indenter. Three indentations were made at 100 µm and 500 µm from the root canal lumen. Afterwards, the specimens were prepared for scanning electron microscopic analysis and the amount of dentin erosion was examined. Wilcoxon and Kruskal-Wallis tests were used to analyze the results with a significance level set at 5%. At 100 µm, all protocols significantly reduced dentin microhardness (p < .05), while at 500 µm, this effect was detected only in the EDTA and QMiX groups (p < .05). CA was the irrigant that caused more extensive erosion in dentinal tubules, followed by PA and EDTA. QMiX opened dentinal tubules, but did not cause dentin erosion. Results suggest that QMiX and 17% EDTA reduced dentin microhardness at a greater depth. Additionally, QMiX did not cause dentin erosion.

Stability of antimicrobial activity of peracetic acid solutions used in the final disinfection process

The instruments and materials used in health establishments are frequently exposed to microorganism contamination, and chemical products are used before sterilization to reduce occupational infection. We evaluated the antimicrobial effectiveness, physical stability, and corrosiveness of two commercial formulations of peracetic acid on experimentally contaminated specimens. Stainless steel specimens were contaminated with Staphylococcus aureus, Escherichia coli, Candida albicans, blood, and saliva and then immersed in a ready peracetic acid solution: 2% Sekusept Aktiv (SA) or 0.25% Proxitane Alpha (PA), for different times. Then, washes of these instruments were plated in culture medium and colony-forming units counted. This procedure was repeated six times per day over 24 non-consecutive days. The corrosion capacity was assessed with the mass loss test, and the concentration of peracetic acid and pH of the solutions were measured with indicator tapes. Both SA and PA significantly eliminated microorganisms; however, the SA solution was stable for only 4 days, whereas PA remained stable throughout the experiment. The concentration of peracetic acid in the SA solutions decreased over time until the chemical was undetectable, although the pH remained at 5. The PA solution had a concentration of 500-400 mg/L and a pH of 2-3. Neither formulation induced corrosion and both reduced the number of microorganisms (p = 0.0001). However, the differences observed in the performance of each product highlight the necessity of establishing a protocol for optimizing the use of each one.

Biofilm formation by Staphylococcus aureus from food contact surfaces in a meat-based broth and sensitivity to sanitizers

This study assessed the capacity of adhesion, the detachment kinetic and the biofilm formation by Staphylococcus aureus isolated from food services on stainless steel and polypropylene surfaces (2 x 2 cm) when cultivated in a meat-based broth at 28 and 7 ºC. It was also to study the efficacy of the sanitizers sodium hypochlorite (250 mg/L) and peracetic acid (30 mg/L) in inactivating the bacterial cells in the preformed biofilm. S. aureus strains adhered in high numbers regardless the assayed surface kind and incubation temperature over 72 h. Cells detachment of surfaces revealed high persistence over the incubation period. Number of cells needed for biofilm formation was noted at all experimental systems already after 3 days. Peracetic acid and sodium hypochlorite were not efficient in completely removing the cells of S. aureus adhered on polypropylene and stainless steel surfaces. From these results, the assayed strains revealed high capacity to adhere and form biofilm on polypropylene and stainless steel surfaces under different growth conditions. Moreover, the cells in biofilm matrix were resistant for total removal when submitted to the exposure to sanitizers.

Alternative sanitization methods for minimally processed lettuce in comparison to sodium hypochlorite

Lettuce is a leafy vegetable widely used in industry for minimally processed products, in which the step of sanitization is the crucial moment for ensuring a safe food for consumption. Chlorinated compounds, mainly sodium hypochlorite, are the most used in Brazil, but the formation of trihalomethanes from this sanitizer is a drawback. Then, the search for alternative methods to sodium hypochlorite has been emerging as a matter of great interest. The suitability of chlorine dioxide (60 mg L-1/10 min), peracetic acid (100 mg L-1/15 min) and ozonated water (1.2 mg L-1 /1 min) as alternative sanitizers to sodium hypochlorite (150 mg L-1 free chlorine/15 min) were evaluated. Minimally processed lettuce washed with tap water for 1 min was used as a control. Microbiological analyses were performed in triplicate, before and after sanitization, and at 3, 6, 9 and 12 days of storage at 2 ± 1 ºC with the product packaged on LDPE bags of 60 µm. It was evaluated total coliforms, Escherichia coli, Salmonella spp., psicrotrophic and mesophilic bacteria, yeasts and molds. All samples of minimally processed lettuce showed absence of E. coli and Salmonella spp. The treatments of chlorine dioxide, peracetic acid and ozonated water promoted reduction of 2.5, 1.1 and 0.7 log cycle, respectively, on count of microbial load of minimally processed product and can be used as substitutes for sodium hypochlorite. These alternative compounds promoted a shelf-life of six days to minimally processed lettuce, while the shelf-life with sodium hypochlorite was 12 days.

Effectiveness of 2 percent peracetic acid for the disinfection of gutta-percha cones

The aim of this study was to evaluate the effectiveness of 2 percent peracetic acid for the disinfection of gutta-percha cones contaminated in vitro with Escherichia coli, Staphylococcus aureus, Streptococcus mutans, Candida albicans and Bacillus subtilus (in spore form). Two hundred and twenty-five gutta-percha cones were contaminated with standardized suspensions of each microorganism and incubated at 37°C for 24 h. The cones were divided into 10 experimental groups (n = 15), according to the microorganism tested and disinfection testing times. The disinfection procedure consisted of immersing each cone in a plastic tube containing the substance. The specimens remained in contact with the substance for 1 or 2.5 minutes. Afterwards, each cone was transferred to a 10 percent sodium thiosulphate solution (Na2S2O3) to neutralize the disinfectant. Microbial biofilms adhering to the cones were dispersed by agitation. Aliquots of 0.1 ml of the suspensions obtained were plated on Sabouraud dextrose agar, or brain and heart infusion agar, and incubated at 37°C for 24 h. The results were expressed in colony forming units (CFU/ml) and the data were submitted to the Wilcoxon Signed Rank Test (level of significance at 0.05). A significant reduction was observed, after 1 minute of exposure, in the test solution for C. albicans (p = 0.0190), S. aureus (p = 0.0001), S. mutans (p = 0.0001), B. subtilis (p = 0.0001), and E. coli (p = 0.0001). After 2.5 minutes of exposure, 100 percent of the microbial inocula were eliminated. It was concluded that the 2 percent peracetic acid solution was effective against the biofilms of the tested microorganisms on gutta-percha cones at 1 minute of exposure.

Antibacterial effectiveness of peracetic acid and conventional endodontic irrigants

This study evaluated the in vitro antibacterial activity of conventional and experimental endodontic irrigants against Enterococcus faecalis. The following substances were evaluated by direct contact test: 2.5 percent sodium hypochlorite (NaOCl); 2 percent chlorhexidine (CHX); 1 percent peracetic acid. After different contact periods (30 s, 1, 3, and 10 min), a neutralizing agent was applied. Serial 10-fold dilutions were prepared and plated onto tryptic soy agar (TSA) and the number of colony-forming units per milliliter (CFU/mL) was determined. Sterile saline was used as a negative control. Both 2.5 percent NaOCl and 2 percent CHX eliminated E. faecalis after 30 s of contact. Peracetic acid reduced the bacterial counts by 86 percent after 3 min and completely eliminated E. faecalis after 10 min. These results allow us to conclude that 1 percent peracetic acid is effective against E. faecalis, despite its slower action compared with 2.5 percent NaOCl and 2 percent CHX.

Este estudo avaliou, in vitro, a atividade antibacteriana de soluções irrigadoras convencionais e experimentais sobre Enterecoccus faecalis. As seguintes substâncias foram avaliadas por teste de contato direto: hipoclorito de sódio (NaOCl) a 2,5 por cento, clorexidina (CHX) a 2 por cento, ácido peracético a 1 por cento. Após diferentes períodos de contato (30 s, 1, 3 e 10 min), um agente neutralizante foi empregado. Diluições decimais seriadas foram realizadas e semeadas em placas de tryptic soy agar (TSA). O número de unidades formadoras de colônia por mililitro (UFC/mL) foi determinado. Solução salina foi utilizada como controle negativo. Ambos, NaOCl a 2,5 por cento e CHX a 2 por cento, eliminaram E. faecalis após 30 s de contato. O ácido peracético reduziu a contagem bacteriana em 86 por cento após 3 min e eliminou completamente E. faecalis após 10 min. Estes resultados permitem concluir que o ácido peracético a 1 por cento é efetivo sobre E. faecalis, apesar de sua ação mais lenta quando comparado ao NaOCl a 2,5 por cento e CHX a 2 por cento.

Microbiological evaluation of peracetic acid for disinfection of acrylic resins / Avaliação microbiológica de ácido peracético na desinfecção de resinas acrílicas

PURPOSE: The aim of this study was to assess the efficacy of peracetic acid (PAA) for the disinfection of dental acrylic resins experimentally contaminated with Candida albicans, Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa. METHODS: Fifteen materials were used for each type of resin (thermosetting, self-curing and microwave-curing). Each material was placed in a test tube containing culture medium with a suspension of each microorganism and then incubated. The materials were rinsed and transferred to other tubes containing 50 mL of water for 5 min, 0.2 percent peracetic acid for 5 min or glutaraldehyde for 30 min. The materials were placed in the culture agar and incubated. Microbial growth was determined by colony counting after plating. RESULTS: Candida albicans growth was inhibited by peracetic acid and glutaraldehyde treatments. The number of colonies on resins treated with saline was greater than 10(5) CFU/mL. In resins infected with E. coli, S. aureus and P. aeruginosa the colony growth was not inhibited by saline and peracetic acid, but it was totally inhibited by glutaraldehyde. CONCLUSION: Surface disinfection using peracetic acid effectively inhibited C. albicans growth on all acrylic resins.

OBJETIVO: O objetivo deste estudo foi avaliar a eficácia do ácido peracético (PAA) na desinfecção de resinas acrílicas dentais experimentalmente contaminadas com Candida albicans, Escherichia coli, Staphylococcus aureus e Pseudomonas aeruginosa. METODOLOGIA: Quinze corpos de prova (CP) foram utilizados para cada tipo de resina (termopolimerizáveis, autopolimerizáveis e ativados por energia de micro-ondas). Cada CP foi colocado em um tubo teste com meio de cultura contendo uma suspensão de cada microrganismo e incubado. Posteriormente, os CP foram lavados e transferidos para outros tubos contendo 50 mL de água por 5 minutos, em 0,2 por cento de ácido peracético por 5 min ou em glutaraldeído por 30 minutos, plaqueados em ágar de cultura e incubados. O crescimento microbiano foi determinado por contagem de colônias após o plaqueamento. RESULTADO: O crescimento de Candida albicans foi inibido nos tratamentos com ácido peracético e glutaraldeído. O número de colônias nas resinas tratadas com solução salina foi superior a 10(5) UFC/mL. Nas resinas infectadas com E. coli, S. aureus e P. aeruginosa, o crescimento das colônias não foi inibido nas resinas tratadas com salina e ácido peracético, mas foi totalmente inibida pelo glutaraldeído. CONCLUSÃO: A desinfecção com ácido peracético inibiu efetivamente o crescimento de C. albicans em todas as resinas acrílicas.

Mycobacterium massiliense BRA100 strain recovered from postsurgical infections: resistance to high concentrations of glutaraldehyde and alternative solutions for high level disinfection / Mycobacterium massiliense clone BRA100 associado a infecções pós-cirúrgicas: resistência a altas concentrações de glutaraldeído e produtos alternativos para desinfecção de alto nível

PURPOSE: To evaluate the minimum inhibitory concentration (MIC) of GTA against these microorganisms and alternative disinfectants for high-level disinfection (HLD). METHODS: Reference mycobacteria and clinical M. massiliense strains were included in this study. Active cultures were submitted to susceptibility qualitative tests with GTA dilutions (ranging from 1.5 percent to 8 percent), and commercial orthophthaldehyde (OPA) and peracetic acid (PA) - based solutions, during the period of exposure as recommended by National Agency of Sanitary Surveillance for HLD. RESULTS: All reference and M. massiliense non-BRA100 strains, recovered from sputum, were susceptible to any GTA concentration, OPA and PA solutions. M. massiliense BRA100 strains presented MIC of 8 percent GTA and were susceptible to OPA and PA. CONCLUSION: M. massiliense BRA100 strain is resistant to high GTA concentrations (up to 7 percent), which proves that this product is non-effective against specific rapidly growing mycobacteria and should be substituted by OPA or PA - based solutions for HLD.

OBJETIVO: Avaliar a concentração mínima inibitória (CMI) de GTA frente a M. massiliense e a susceptibilidade a produtos alternativos para desinfecção de alto nível (DAN). MÉTODOS: Cepas de M. massiliense de origem clínica e de referência foram incluídas no estudo. As culturas ativadas foram submetidas a testes qualitativos com diluições de GTA (de 1,5 por cento a 8 por cento) e com soluções comerciais de ortoftaldeído (OPA) ou ácido peracético (PA), utilizando os tempos de exposição recomendados pela Agência Nacional de Vigilância Sanitária para DAN. RESULTADOS: Todas as cepas de referência e M. massiliense não-BRA100, obtida de escarro, foram susceptíveis às concentrações de GTA, e soluções de OPA e PA. As cepas de M. massiliense BRA100 apresentaram CMI de 8 por cento para GTA e foram susceptíveis a OPA e PA. CONCLUSÃO: M. massiliense BRA100 é resistente a altas concentrações de GTA (até 7 por cento), o que demonstra que esse composto não é eficaz, e deve ser substituído por OPA ou PA nos processos de DAN.

Evaluation of the effectiveness of peracetic acid in the sterilization of dental equipment.

PURPOSE: To evaluate the effectiveness of peracetic acid in the microbiological sterilisation of dental materials. METHODS: Peracetic acid solution was evaluated at concentrations of 800, 1500 and 2500 ppm. At these concentrations, it was determined whether peracetic acid caused corrosion to dental instruments and induced cellular mutagenicity and cytotoxicity. In addition, the minimum inhibitory concentration (MIC), the minimum bactericidal concentration (MBC), agar diffusion and diffusion by well method, were also verified. RESULTS: The corrosion rate, calculated from potentiodynamic assays was 10(-6) cm/year, indicating that the product does not damage equipment. The sterilisation capacity of peracetic acid at 2500 ppm was the best. The comet assay indicated genotoxic activity at 2500 ppm. CONCLUSIONS: This study demonstrated the effectiveness of peracetic acid for sterilizing dental equipment, providing another alternative for the prevention of infections in clinics.

In Vivo and In Vitro evaluation of the efficacy of a peracetic acid-based disinfectant for decontamination of acrylic resins

O objetivo deste trabalho foi avaliar a eficácia antimicrobiológica de um desinfetante à base de ácido peracético na descontaminação de resinas acrílicas termicamente ativada, quimicamente ativada e polimerizada em forno de microondas. Placas de resina foram contaminadas in vivo por meio do uso intraoral por 10 voluntários durante 7 noites e corpos-de-prova de resina foram contaminados in vitro por meio do contato com microrganismos conhecidos: Bacillus subtilis e Bacillus stearothermophilus. Os espécimes contaminados foram imersos em desinfetante à base de ácido peracético a 0,2% (Sterilife®; Lifemed) durante 5 ou 10 min e então colocados no meio de cultura BHI. Após incubação a 37°C for 48 h, o crescimento bacteriano foi avaliado por meio análise da turvação do meio de cultura. Todos os espécimes imersos em ácido peracético por 5 ou 10 min não apresentaram turvação do meio de cultura, enquanto os espécimes contaminados e colocados diretamente no meio de cultura (grupo controle) apresentaram turvação. Concluiu-se que a imersão em ácido peracético por pelo menos 5 min foi eficaz na desinfecção de resinas acrílicas termicamente ativada, quimicamente ativada e polimerizada em forno de microondas contaminadas tanto com saliva humana quanto com Bacillus subtilis ou Bacillus stearothermophilus.

Tratamientos de desinfección de lechugas (Lactuca sativa) y frutillas (Fragaria chiloensis) / Disinfection treatment for lettuces (Lactuca sativa) and strawberries (Fragatia Chiloensis)

The disinfection of vegetables and fruits is a treatment applied in order to reduce their natural contamination or processes to the product along the different steps of the food chain until its consumption. In the present work the effect of two disinfectants products was studied: grapefruit seed extract (400 ppm) for 10 min and peracetic acid (2000 ppm) for 1 min (action times assayed according to the manufacturer recommendations) and other additional times. The germicidal action was carried out against the natural contaminants of lettuces and strawberries, through the determination of the germicidal efficiency (). Treated and untreated strawberries were also evaluated for flavor changes through a sensorial difference test, triangular test. None of the assayed products reached the 99.999 destruction of the natural contaminants according to the Chambers test. Peracetic acid was the most effective disinfectant, reaching the highest destruction percentages at a time lower than that for grapefruit seed extract. Sensory analysis showed no significant differences (p = .05) between strawberries with and without disinfection treatments, at the conditions suggested by the manufacturer.

Microbial killing activity of peracetic acid.

In vitro killing activity of peracetic acid (Perasafe) at a concentration of 0.26 per cent w/v was tested against Escherichia coli, Enterobacter cloacae, Klebsiella pneumoniae, Pseudomonas aeruginosa, Salmonella typhi, Salmonella paratyphi A, Acinetobacter baumannii, Sternotrophomonas maltophilia, Enterococcus faecium, Enterococcus faecalis, methicillin-resistant Staphylococcus aureus (MRSA), Bacillus subtilis spore, Mycobacterium tuberculosis and human immuno-deficiency virus type I. Exposure to Peracetic acid (0.26% w/v) for 10 minutes resulted in massive killing of all the aforementioned organisms and spore.