Susceptibility of clinical isolates of multiresistant Pseudomonas aeruginosa to a hospital disinfectant and molecular typing

The aim of this study was to evaluate the susceptibility of 35 resistant Pseudomonas aeruginosa clinical isolates to a quaternary ammonium hospital disinfectant. The methodology was the AOAC Use-Dilution Test, with disinfectant at its use-concentration. In addition, the chromosomal DNA profile of the isolates were determined by macro-restriction pulsed field gel electrophoresis (PFGE) method aiming to verify the relatedness among them and the behavior of isolates from the same group regarding the susceptibility to the disinfectant. Seventy one percent of the isolates were multiresistant to antibiotics and 43% showed a reduced susceptibility to the disinfectant. The PFGE methodology detected 18 major clonal groups. We found isolates with reduced susceptibility to the disinfectant and we think that these are worrying data that should be further investigated including different organisms and chemical agents in order to demonstrate that microorganisms can be destroyed by biocide as necessary. We also found strains of the same clonal groups showing different susceptibility to the disinfectant. This is an interesting observation considering that only few works are available about this subject. PFGE profile seems not to be a reliable marker for resistance to disinfectants.

Hand-disinfectant alcoholic vapors in incubators

Objective: To analyze the atmosphere inside incubators regarding alcoholic solvent such as isopropanol or ethanol which are commonly used in hand disinfecting solutions. Design: Observational. Setting: The third level neonatal unit of the Centre Hospitalier Universitaire Vaudois, Lausanne, Switzerland. Patients: Nine neonates with median (range) gestational age of 29 4/7 (25 5/7-39 0/7) weeks and birth weight of 960 (550-3050) grams. All neonates were inside incubators. Interventions: Alcoholic vapors inside incubators were directly and cumulatively measured by photoionisation and gas chromatography respectively after absorption on a charcoal sampling tube. Results: Eleven studies (mean study time: 230 ± 19 minutes) were performed. Highly variable isopropanol/ethanol concentrations profiles were found inside incubators. Peak value for isopropanol was 1982 part per million and for ethanol was 906 part per million. Conclusions: Incubators' inner atmosphere can be highly polluted by alcohol vapors. To reduce them staff should respect long evaporation time between hands disinfection and manipulations inside incubators. The use of an ethanol-based disinfecting solution, because of its short evaporation time, could be favored. As alcohol vapor toxicity for neonate remains largely unknown, further studies could be welcome.

Hand-disinfectant alcoholic vapors in incubators

La désinfection des mains avant d'effectuer des soins est un acte indispensable en médecine clinique pour limiter le risque de transmission de germes. Après utilisation des produits désinfectants mis à disposition dans les services de soins, il se dégage une odeur alcoolique forte et désagréable, liée directement aux alcools antimicrobiens des solutions. Une étude a montré qu'une exposition aiguë et brève aux vapeurs d'éthanol et isopropanol chez des enfants prématurés pouvait être mise en relation avec des changements hémodynamiques au niveau de la zone olfactive orbito-frontale [1]. Aucune norme réglementant les concentrations de vapeurs d'éthanol ou isopropanol auxquelles les nouveau-nés peuvent être exposés n'existe. Cette thèse avait pour but d'étudier l'exposition des nouveaux nés soignés dans des incubateurs à des vapeurs d'alcool (éthanol et isopropanol). Elle était composée de 2 parties qui ont été publiées dans 2 articles différents et qui représentent le travail de doctorat [2-3]. La 1ère partie était une étude observationnelle d'une série de cas [2], Des mesures des concentrations des vapeurs d'alcool ont été effectuées auprès de 9 nouveau-nés soignés dans des incubateurs de même modèle au sein de l'unité de néonatologie du Centre Hospitalier Universitaire Vaudois à Lausanne. Sur 4 heures, les concentrations instantanées et moyennes ont été mesurées par deux techniques (photoionisation et respectivement chromatographic après absorption sur charbon actif). Onze analyses ont été effectuées en 2004-2005. Elles ont révélé des taux très variables d'éthanol et d'isopropanol dans les incubateurs (avec des valeurs maximales de 1982 ppm pour l'isopropanol et 906 ppm pour l'éthanol) correspondant aux introduction de mains fraîchement désinfectées dans les isolettes. Les concentrations moyennes variaient entre 9.8 ppm et plus de 61 ppm pour l'éthanol et < 0.01 ppm et 119 ppm pour l'isopropanol. La 2e partie a été réalisée en collaboration avec le PD Dr D. Vernez de l'Institut Universitaire Romand de Santé au Travail [3], Un modèle théorique prédictif des concentrations alcooliques dans des incubateurs pour nouveau-nés a été développé. Des séries de mesures standardisées des variations des concentrations alcooliques dans un incubateur sans patient ont été effectuées en changeant trois variables: 1) le renouvellement de l'air dans l'incubateur en variant le nombre de portes ouvertes, 2) la quantité de solution alcoolique versée sur les mains avant de les introduire dans l'incubateur 3) le temps de séchage des mains après désinfection et avant de les introduire dans l'incubateur. La modélisation a permis de décrire la cinétique des concentrations d'alcool dans les incubateurs et d'évaluer les pistes potentielles pour diminuer les risques d'exposition des nouveau-nés à ces vapeurs dans leurs incubateurs. En conclusion, la 1 ère partie a mis en évidence, pour la première fois, que des nouveau- nés soignés en incubateurs peuvent être exposés à des vapeurs d'alcool. Comme il η y a aucune norme d'exposition pour cette population et que les seules limites d'exposition existantes sont destinées à des travailleurs adultes, aucune conclusion précise ne peut être avancée sur les risques toxicologiques. L'exposition à des vapeurs polluantes d'un nouveau-né à terme ou prématuré, en plein développement neuro-sensoriel, devrait toutefois, à priori, être évitée. La 2ème partie permet de proposer des pistes pratiques pour diminuer les concentrations des vapeurs d'alcool dans les incubateurs: respecter le temps de séchage des mains après leur désinfection et avant de les introduire dans les isolettes et/ou préférer un désinfectant alcoolique à faible temps d'évaporation.

Fitness and dissemination of disinfectant-selected multiple-antibiotic-resistant (MAR) strains of Salmonella enterica serovar Typhimurium in chickens

Objectives: The aims of this study were to determine whether strains of Salmonella enterica serovar Typhimurium which had acquired low-level multiple antibiotic resistance (MAR) through repeated exposure to farm disinfectants were able to colonize and transmit between chicks as easily as the parent strain and, if such strains were less susceptible to fluoroquinolones, would high-level resistance be selected after fluoroquinolone treatment. Methods: Two mutants were compared with the isogenic parent. In the first experiment, day-old chicks were co-infected with both the parent and a mutant to determine their relative fitness. In the second experiment, parent and mutant strains (in separate groups of chicks) were assessed for their ability to transmit from infected (contact) to non-infected (naive) birds and with respect to their susceptibility to fluoroquinolone treatment. Birds were regularly monitored for the presence of Salmonella in caecal contents. Replica plating was used to monitor for the selection of antibiotic-resistant strains. Results: The parent strain was shown to be significantly fitter than the two mutants and was more rapidly disseminated to naive birds. Antibiotic treatment did not preferentially select for the two mutants or for resistant strains. Conclusions: The disinfectant-exposed strains, although MAR, were less fit, less able to disseminate than the parent strain and were not preferentially selected by therapeutic antibiotic treatment. As such, these strains are unlikely to present a greater problem than other salmonellae in chickens.

Effect of triclosan or a phenolic farm disinfectant on the selection of antibiotic-resistant Salmonella enterica

Objective: To determine the effect of growth of five strains of Salmonella enterica and their isogenic multiply antibiotic-resistant (MAR) derivatives with a phenolic farm disinfectant or triclosan (biocides) upon the frequency of mutation to resistance to antibiotics or cyclohexane. Methods: Strains were grown in broth with or without the biocides and then spread on to agar containing ampicillin, ciprofloxacin or tetracycline each at 4x MIC or agar overlaid with cyclohexane. Incubation was for 24 and 48 h and the frequency of mutation to resistance was calculated for strains with and without prior growth with the biocides. MICs were determined and the presence of mutations in the acrR and marR regions was determined by sequencing and the presence of mutations in gyrA by light-cycler analysis, for a selection of the mutants that arose. Results: The mean frequency of mutation to antibiotic or cyclohexane resistance was increased similar to10- to 100-fold by prior growth with the phenolic disinfectant or triclosan. The increases were statistically significant for all antibiotics and cyclohexane following exposure to the phenolic disinfectant (P less than or equal to 0.013), and for ampicillin and cyclohexane following exposure to triclosan (P less than or equal to 0.009). Mutants inhibited by >1 mg/L ciprofloxacin arose only from strains that were MAR. Reduced susceptibility to ciprofloxacin (at 4x MIC for parent strains) alone was associated with mutations in gyrA. MAR mutants did not contain mutations in the acrR or marR region. Conclusions: These data renew fears that the use of biocides may lead to an increased selective pressure towards antibiotic resistance.

Antimicrobial activity of disinfectant agents incorporated into type IV dental stone

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The effect of disinfectant solutions on the hardness of acrylic resin denture teeth

This investigation studied the effects of disinfectant solutions on the hardness of acrylic resin denture teeth. The occlusal surfaces of 64 resin denture teeth were ground flat with abrasives up to 400-grit silicon carbide paper. Measurements were made after polishing and after the specimens were stored in water at 37 degreesC for 48 h. The specimens were then divided into four groups and immersed in chemical disinfectants (4% chlorhexidine; 1% sodium hypochlorite and sodium perborate) for 10 min. The disinfection methods were performed twice to simulate clinical conditions and hardness measurements were made. Specimens tested as controls were immersed in water during the same disinfection time. Eight specimens were produced for each group. After desinfection procedures, testing of hardness was also performed after the samples were stored at 37 degreesC for 7, 30, 60, 90 and 120 days. Data were analysed using two-way analysis of variance (anova) and Tukey's test at 95% confidence level. According to the results, no significant differences were found between materials and immersion solutions (P > 0.05). However, a continuous decrease in hardness was noticed after ageing (P < 0.05). It was conclude that the surfaces of both acrylic resin denture teeth softened upon immersion in water regardless the disinfecting solution.

Effect of Incorporation of Disinfectant Solutions on Setting Time, Linear Dimensional Stability, and Detail Reproduction in Dental Stone Casts

Purpose: The aim of this paper was to analyze the influence of incorporation of disinfectants during the cast die stone-setting time. Setting time, linear dimensional stability, and reproduction details on casts were measured.Materials and Methods: Die stone type IV specimens with disinfection solutions (sodium hypochlorite 1%, glutaraldehyde 2%, chlorhexidine 2%) were incorporated in two concentrations (50%, 100%). The detail reproduction, dimensional stability, and setting time were tested in accordance with ADA recommendations.Results: Disinfecting solutions promoted an increase in setting time compared to control; sodium hypochlorite was responsible for the highest setting time. The addition of undiluted sodium hypochlorite 1.0% led to contraction during setting, but the groups with 50% diluted sodium hypochlorite 1.0% and undiluted chlorhexidine 2.0% resulted in intermediate values compared to the other groups, thus matching the control. The others did not demonstrate any effect on expansion. For detail reproduction, it was observed that the control group presented results similar to the others, except those where sodium hypochlorite was added.Conclusions The addition of sodium hypochlorite in both dilutions significantly altered, negatively, all the evaluated properties. But the addition of glutaraldehyde and chlorhexidine did not promote any significant alterations in the evaluated properties.

Effectiveness of two disinfectant solutions and microwave irradiation in disinfecting complete dentures contaminated with methicillin-resistant Staphylococcus aureus

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of disinfectant agents on dimensional stability of elastomeric impression materials.

STATEMENT OF PROBLEM: Difficulties in sterilizing impressions by traditional methods have led to chemical disinfection as an alternative, and some studies have shown that disinfectants may adversely affect impressions. PURPOSE: This study investigated the effect of disinfection methods on the dimensional stability of 6 elastomeric materials. MATERIAL AND METHODS: Impression materials were submitted to the following treatments: immersion in 5.25% sodium hypochlorite solution for 10 minutes, immersion in 2% glutaraldehyde solution for 30 minutes, and no immersion (control). After treatments, impressions were poured, and respective stone casts were measured with a Nikon Profile projector and compared with the master model. RESULTS: The elastomeric materials had different reproduction capacities, and the disinfecting treatments did not differ from the control.

A bacteriological study of hospital beds before and after disinfection with phenolic disinfectant

In hospitals, one of the ways to control microbial contamination is by disinfecting the furniture used by patients. This study's main objective was to evaluate the microbiological condition of hospital mattresses before and after such disinfection, in order to identify bacteria that are epidemiologically important in nosocomial infection, such as Staphylococcus aureus and Pseudomonas aeruginosa. RODAC plates with two different culture media were used to collect specimens. Patient beds were selected according to previously established criteria, and surface areas on the mattresses were chosen at random. From the total of 1 040 plate cultures from 52 mattresses, positive results were obtained from 500 of them (48.1%), 263 before disinfection and 237 after disinfection. Considering the selectivity of the culture media, the positivity rate was high. There were high prevalences of S. aureus both before and after mattress disinfection. The study results suggest that the usual disinfection procedures, instead of diminishing the number of microbes, merely displace them from one part of the mattress to another, and the number of microorganisms remains the same.

Smear layer removal capacity of disinfectant solutions used with and without EDTA for the irrigation of canals: a SEM study.

The purpose of this study was to carry out a scanning electron microscopic (SEM) analysis of the cleaning qualities and smear layer removal from root canal walls, instrumented and irrigated with 2.5% NaOCl, 2.0% chlorhexidine and saline solutions. Fifty extracted teeth were used in this study. All teeth were radiographed to determine the existence of a single canal. The crowns were cut at the cervical limit and the root canals were instrumented with K-type files up to size 45. During root canal preparation, irrigations were made with the different solutions being evaluated: Group 1: 2.5% NaOCl (10 roots); Group 2: 2.5% NaOCl and 17% EDTA for 2 minute (10 roots); Group 3: 2.0% chlorhexidine (10 roots); Group 4: 2.0% chlorhexidine and 17% EDTA for 2 minutes (10 roots); Group 5: saline solution (5 roots); Group 6: saline solution and 17% EDTA for 2 minutes (5 roots). After instrumentation, the canals were irrigated with each one of the solutions and the roots were cut in the buccolingual direction for SEM analysis, at the cervical, middle and apical thirds, to ascertain the presence or absence of smear layer and debris. SEM analysis was performed by three calibrated examiners and scores were submitted to Kruskal-Wallis test at the significance level of p = 5%. Results showed that the use of 17% EDTA decreased the smear layer significantly (p < 0.05) for all evaluated solutions in all thirds. When EDTA was not used, a significantly higher quantity of smear layer on the apical third was observed only in the NaOCl groups. The use of 17% EDTA was significant for debris removal except for the chlorhexidine groups. The following conclusion could be drawn: the use of 17% EDTA was necessary to enhance cleanness of the root canals.

Effect of disinfectant solutions on a denture base acrylic resin.

The aim of this study was to evaluate the hardness, roughness and mass loss of an acrylic denture base resin after in vitro exposure to four disinfectant solutions. Forty specimens (Clássico, Brazil) were prepared and randomly assigned to 4 groups n = 10) according to the disinfectant solution: G1: control, stored in distilled water at 37 degrees C; G2: 1% sodium hypochlorite; G3: 2% glutaraldehyde; G4: 4% chlorhexidine. G2 to G4 were immersed for 60 minutes in the disinfectant solution. Measurements were carried out both before and after immersion in the solution. The surface was analyzed with a surface roughness tester (Surfcorder SE 1700 KOZAKALAB), a microdurometer FM-700 (Future Tech) and a scanning electron microscope (DSM 962-ZEISS). Loss of mass was determined with a digital weighing scale. After disinfection procedures, values were analyzed statistically. The acrylic denture base resin may be vulnerable to surface changes after in vitro immersion in the disinfectant solutions studied.

Bond strength of artificial teeth attached to a microwave-polymerized denture base resin after immersion in disinfectant solutions

To evaluate the bond strength between two types of acrylic resin teeth and a microwave denture base resin after immersion in disinfectant solutions for 180 days. Eighty specimens made of acrylic resin teeth (Biotone and Biotone IPN) attached to a microwave polymerized denture base resin (Nature-Cryl MC) were divided into eight groups (n = 10) according to the treatment (distilled water-control, 2% chlorhexidine digluconate, 1% sodium hypochlorite and sodium perborate solution-Corega Tabs). The shear strength tests (MPa) were carried out using a universal testing machine with a 0.5 mm/min speed. Data analysis was performed using ANOVA and multiple comparison Student-Newman-Keuls post hoc test (α = 0.05). Biotone IPN showed similar results among the groups (distilled water, 8.25 ± 1.81 MPa; chlorhexidine, 7.81 ± 3.34 MPa; hypochlorite, 7.75 ± 3.72 MPa; and Corega Tabs, 7.58 ± 2.27 MPa, whereas Biotone showed significantly lower shear bond strength values for the groups immersed in Corega Tabs (5.25 ± 3.27 MPa) and chlorhexidine (6.08 ± 2.35 MPa). Soaking the dentures in 1% sodium hypochlorite could be recommended as a disinfectant solution for dentures fabricated with conventional acrylic resin denture teeth and microwave denture base resin. For dentures fabricated with IPN teeth and microwave denture base resin, all the soaking solutions evaluated in this study could be suggested to denture wearers.

Copper as an algicide and disinfectant in water supplies.

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