Assessment of three antibiotic combination regimens against Gram-negative bacteria causing neonatal sepsis in low- and middle-income countries.

Gram-negative bacteria (GNB) are a major cause of neonatal sepsis in low- and middle-income countries (LMICs). Although the World Health Organization (WHO) reports that over 80% of these sepsis deaths could be prevented through improved treatment, the efficacy of the currently recommended first- and second-line treatment regimens for this condition is increasingly affected by high rates of drug resistance. Here we assess three well known antibiotics, fosfomycin, flomoxef and amikacin, in combination as potential antibiotic treatment regimens by investigating the drug resistance and genetic profiles of commonly isolated GNB causing neonatal sepsis in LMICs. The five most prevalent bacterial isolates in the NeoOBS study (NCT03721302) are Klebsiella pneumoniae, Acinetobacter baumannii, E. coli, Serratia marcescens and Enterobacter cloacae complex. Among these isolates, high levels of ESBL and carbapenemase encoding genes are detected along with resistance to ampicillin, gentamicin and cefotaxime, the current WHO recommended empiric regimens. The three new combinations show excellent in vitro activity against ESBL-producing K. pneumoniae and E. coli isolates. Our data should further inform and support the clinical evaluation of these three antibiotic combinations for the treatment of neonatal sepsis in areas with high rates of multidrug-resistant Gram-negative bacteria.

Genotyping of paired KPC-producing Klebsiella pneumoniae isolates with and without divergent polymyxin B susceptibility profiles.

Polymyxins are still used mainly in treating infections caused by carbapenem-resistant Klebsiella pneumoniae worldwide. The most frequent mechanism of acquired resistance to polymyxins in Gram-negative bacilli is the occurrence of mutations in chromosomal genes regulating operons responsible for lipopolysaccharide modification. As we observed at Santa Casa de São Paulo hospital the occurrence of infections caused by isolates resistant to polymyxins in patients previously treated with this antimicrobial, and new infections caused by the same polymyxin-susceptible species, in this study, we aimed to determine the clonality of consecutive K. pneumoniae isolates from the same patients and characterize the molecular determinants of polymyxin resistance in paired or clonal isolates. A total of 24 pairs and one trio of K. pneumoniae isolates were included in this study. Species identification was achieved by mass spectrometry and multiplex PCR. Polymyxin B minimal inhibitory concentrations were determined by broth microdilution. Clonality was evaluated using pulsed-field gel electrophoresis. The presence of insertions in mgrB gene was tested by PCR, and mutations on pmrA, pmrB, phoP, and phoQ were evaluated by PCR and complete nucleotide sequencing. A fraction of 23.8% of strains resistant to polymyxin B had an insertion in mgrB. Amino acid substitution F204L in PmrB may be implicated in polymyxin resistance. Substitutions T246A and R256G in PmrB were not implicated in polymyxin resistance. In this study, polymyxin resistance after a first susceptible isolate was detected was most frequently due to an infection caused by a distinct clone.

The impact of sonication cultures when the diagnosis of prosthetic joint infection is inconclusive.

BACKGROUND: In the absence of a gold standard criterion for diagnosing prosthetic joint infections (PJI), sonication of the removed implant may provide superior microbiological identification to synovial fluid and peri-implant tissue cultures. The aim of this retrospective study was to assess the role of sonication culture compared to tissue cultures for diagnosing PJI, using different consensus and international guidelines for PJI definition. METHODS: Data of 146 patients undergoing removal of hip or knee arthroplasties between 2010 and 2018 were retrospectively reviewed. The International Consensus Meeting (ICM-2018), Musculoskeletal Infection Society (MSIS), Infectious Diseases Society of America (IDSA), the European Bone and Joint Infection Society (EBJIS), and a modified clinical criterion, were used to compare the performance of microbiological tests. McNemar´s test and proportion comparison were employed to calculate p-value. RESULTS: Overall, 56% (82/146) were diagnosed with PJI using the clinical criteria. Out of these cases, 57% (47/82) tested positive on tissue culture and 93% (76/82) on sonication culture. Applying this clinical criterion, the sensitivity of sonication fluid and tissue cultures was 92.7% (95% CI: 87.1%- 98.3%) and 57.3% (95% CI: 46.6%-68.0%) (p<0.001), respectively. When both methods were combined for diagnosis (sonication and tissue cultures) sensitivity reached 96.3% (95% CI: 91.5%-100%). Sonication culture and the combination of sonication with tissue cultures, showed higher sensitivity rates than tissue cultures alone for all diagnostic criteria (ICM-18, MSIS, IDSA and EBJIS) applied. Conversely, tissue culture provided greater specificity than sonication culture for all the criteria assessed, except for the EBJIS criteria, in which sonication and tissue cultures specificity was 100% and 95.3% (95% CI: 87.8-100%), respectively (p = 0.024). CONCLUSIONS: In a context where diagnostic criteria available have shortcomings and tissue cultures remain the gold standard, sonication cultures can aid PJI diagnosis, especially when diagnostic criteria are inconclusive due to some important missing data (joint puncture, histology).

Short-chain cyanoacrylates and long-chain cyanoacrylates (Dermabond) have different antimicrobial effects.

OBJECTIVE: To compare the antimicrobial effect in vitro of a short-chain cyanoacrylate with a long-chain cyanoacrylate (Dermabond, Ethicon, Johnson and Johnson, USA) against bacterial strains. METHODS AND ANALYSIS: The following bacterial strains were analysed: Staphylococcus aureus, Escherichia coli, Klebsiella pneumonia and Pseudomonas aeruginosa. For each microorganism, standardised sterile discs (6 mm) containing 10 µL of ethyl-cyanoacrylate and 2-octyl cyanoacrylate were applied to the plate. All plates received a blank filter-paper disc with no adhesive (control). All plates were incubated for 24 hours, after which the bacterial inhibitory halos, if present, were measured in millimetres in its greater length. RESULTS: Inhibitory halos were observed for both adhesives for S. aureus. Inhibition halos were observed only for ethyl-cyanoacrylate for K. pneumoniae and E. coli. No inhibition halo was observed for P. aeruginosa in any sample. The relationship between the total size of the inhibition halos and the diameter of the paper filter for S. aureus was statistically significant compared with 2-octyl cyanoacrylate. CONCLUSION: Data shown conclude that ethyl-cyanoacrylate showed in vitro bacteriostatic activity for S. aureus, E. coli and K. pneumoniae. 2-Octyl cyanoacrylate showed in vitro lower bacteriostatic activity only against S. aureus when compared with ethyl-cyanoacrylate. No in vitro bactericidal activity of ethyl-cyanoacrylate or 2-octyl cyanoacrylate was observed.

Microbiological diagnosis of intramedullary nailing infection: comparison of bacterial growth between tissue sampling and sonication fluid cultures.

BACKGROUND: Intramedullary nailing (IMN) has been frequently indicated to treat long bone open and closed fractures, but IMN infection (IMNI) may have devastating consequences. Sonication has been regarded as an important add-on for microbial identification on a variety of orthopaedic implant-associated infections, but its role in the IMNI is poorly studied. We aim at evaluating the accuracy obtained by conventional peri-implant tissue culture (TC) samples with sonication fluid cultures (SCs) of IMNI. METHODS: Longitudinal prospective cohort study ongoing since June 2014, which included patients with indication for IMN removal due to any reason. Clinical diagnosis of INMI was defined according to publication addressing fracture-related infections. Minimal of two samples from TC were cultured. SCs followed the protocol previously published. Statistical analysis was performed using McNemar's test for related proportions. RESULTS: We included 54 patients submitted to IMN retrieval, of whom 47 presenting clinical signs of IMNI. Sensitivity for detecting microorganisms using TC and SC was 89.4% (42/47) and 97.6% (40/41), and specificity was 71.4% (5/7) for both TC and SC (p = 1.00). Positive and negative predictive values for TC and SC were 95.5% (42/44), 95.2% (40/42), 50% (5/10), and 83.3% (5/6), respectively. The most frequent organisms isolated in both TC and SC were Staphylococcus aureus, S. epidermidis, and Enterococcus sp. Polymicrobial infection was diagnosed in 14.8% (8/54) and 25% (12/48) by TC and SC, respectively (p = 0.19). CONCLUSION: Sonication fluid and tissue samples presented similar accuracy on the diagnosis of IMNI, but SC was advantageous of detecting polymicrobial infection.

Panton-Valentine Positive Staphylococcus aureus in Community-Acquired and Hospital-Acquired Pediatric Infections.

From July 2009 to July 2015, Staphylococcus aureus isolated from pediatric sterile sites were selected. Polymerase chain reaction was used to detect mecA and lukS-PV/lukF-PV genes. The rate of methicillin-resistant Staphylococcus aureus was 37.7%. Ten isolates had the lukS-PV/lukF-PV genes, 2 of which were methicillin-resistant Staphylococcus aureus. Skin and soft tissues infections were significantly associated with lukS-PV/lukF-PV positive isolates, P = 0.008.

Hospital-Acquired Infections in Children: A Latin American Tertiary Teaching Hospital 5-Year Experience.

Hospital-acquired infections account for high mortality rates and hospital costs. We analyzed pediatric data from a tertiary teaching hospital and found that most of the cases occurred in the intensive care unit and had significant association with invasive devices. Bloodstream infections were the main site of infection, and Gram-negative bacteria were the predominant etiology.

Evaluation of aflibercept and ziv-aflibercept binding affinity to vascular endothelial growth factor, stability and sterility after compounding.

PURPOSE: To investigate the binding affinity, stability, and sterility of aflibercept and ziv-aflibercept to vascular endothelial growth factor (Holash et al. in Proc Natl Acad Sci USA 99(17):11393-11398, 2002. 10.1073/pnas.172398299) after compounding and storage for up to 28 days at 4 °C and - 8 °C. METHODS: Tuberculin-type 1-mL syringes were prepared containing aflibercept (40 mg/mL) and ziv-aflibercept (25 mg/mL). Samples were stored at 4 °C and - 8 °C for 0, 14, and 28 days and evaluated for the binding affinity of anti-VEGF to VEGF and stability using enzyme-linked immunosorbent assays. The evaluation of sample sterility was performed. RESULTS: Laboratory trials with aflibercept and ziv-aflibercept showed preservation of the drug-binding capability to recombinant VEGF when stored in plastic syringes for up to 28 days at 4 °C and - 8 °C. No significant decrease in mass or concentration were observed. Microbiologic evaluations did not detect contamination in the syringes. CONCLUSIONS: The current study corroborates that compounded anti-VEGF drugs aflibercept and ziv-aflibercept do not loose stability or binding affinity and do not become contaminated if prepared under sterile conditions and stored at 4 °C or - 8 °C for 14 or 28 days.

Avaliação da segurança do processamento de fresas intramedulares flexíveis para cirurgia ortopédica / Safety assessment of reprocessing of flexible intramedullary bone reamers for orthopedic surgery / Evaluación de la seguridad del procesamiento de fresas intramedulares flexibles para cirugía ortopédica

Objetivos: Avaliar a eficácia de um procedimento operacional padrão para limpeza de fresas intramedulares flexíveis, bem como o alcance da esterilidade, e evi-denciar a citotoxicidade da sujidade residual de uma fresa flexível utilizada na prática assistencial. Métodos: Fresas intramedulares flexíveis foram pesadas antes do processamento, após contaminação desafio e depois da limpeza. Elas foram contaminadas com Soil Test™, suspensão de Geobacillus stearothermophilus, na concentração de 106 UFC/mL, e farinha de osso bovino. Após processamento, as amostras foram incubadas em meio de cultura por 21 dias. A sujidade residual de uma fresa utilizada na prática foi submetida ao teste de citotoxicidade in vitro. Resultados: As amostras, embora esterilizadas, apontaram acúmulo de sujidade e o processamento foi ineficaz. A sujidade residual apresentou efeito citotóxico. Conclusão: Recomenda-se que o design flexível das fresas seja descontinuado pela insegurança no processamento.

Objectives: To assess the efficacy of a standard operational procedure to clean flexible intramedullary bone reamers, as well as the sterilization level, and to show the cytotoxicity of the residual dirtiness of a flexible reamer used in care practice. Methods: Flexible intramedullary bone reamers were weighed before processing, after challenge contamination and after cleaning. They were contaminated with the Soil Test™, Geobacillus stearothermo-philus suspension, in the concentration of 106 cfu/ml, and bovine bone flour. After processing, the samples were inoculated into a culture medium and incubated for 21 days. Residual dirtiness of a flexible intramedullary bone reamer used in practice was submitted to in vitro cytotoxicity test. Results: Despite being sterilized, the samples indicate to accumulated dirtiness and the processing was inefficient. Residual dirtiness presented a cytotoxic effect. Conclusion: It is recommended that the flexible design of reamers is discontinued by the lack of safety of reprocessing.

Objetivos: Evaluar la eficacia de un procedimiento operacional estándar para limpieza de fresas intramedulares flexibles, así como el alcance de la esterilidad, y evidenciar la citotoxicidad de la suciedad residual de una fresa flexible utilizada en la práctica asistencial. Métodos: Fresas intramedulares flexibles fueron pesadas antes del procesamiento, tras contaminación desafío y después de la limpieza. Fueron contaminadas con Soil Test™, suspensión de Geobacillus stearothermophilus, en la concentración de 106 UFC/mL, y harina de hueso bovino. Tras el procesamiento, las muestras fueron incuba-das en medio de cultura por 21 días. La suciedad residual de una fresa utilizada en la práctica fue sometida al test de citotoxicidad in vitro. Resultados: Las muestras, aunque esterilizadas, apuntaron acumulación de suciedad y el procesamiento fue ineficaz. La suciedad residual presentó efecto citotóxico. Conclusión: Se recomienda que el design flexible de las fresas sea descontinuado por la inseguridad en el procesamiento.

Microbiological evaluation of the steam sterilization of assembled laparoscopic instruments.

Objective: assess the safety of steam sterilization of assembled laparoscopic instruments with challenge contamination. Method: a laboratory experimental study, using as test samples trocars and laparoscopic graspers. Geobacillus stearothermophillus ATCC-7953 was used, with a microbial population of 106UFC/Filter paper substrate, removed from the biological indicator. Three of them were introduced into each instrument at the time of assembly, and sterilized at pressurized saturated steam, 134oC for 5 minutes. After sterilization, the instrument was disassembled and each filter paper substrate was inoculated in soybean casein culture and incubated at 56oC for 21 days. In case of absence of growth, they were subjected to heat shock of 80oC, for 20 minutes and re-incubated for 72 hours. Sample size: 185 graspers and 185 trocars, with 95% power. We paired the experiments with comparative negative control groups (5 graspers and 5 trocars with challenge contamination, sterilized disassembled) and positive control (30 filter paper supports, unsterilized), subject to the same incubation procedures. Results: there was no microbial growth in experimental and negative control. The results of the positive control were satisfactory. Conclusion: this study provided strong scientific evidence to support the safety of steam sterilizing of the assembled laparoscopic instrument.

Microbial investigation of biofilms recovered from endotracheal tubes using sonication in intensive care unit pediatric patients

Abstract Objectives To compare cultured microorganisms identified on endotracheal tubes biofilms through sonication technique with traditional tracheal aspirate collected at extubation of pediatric intensive care unit patients. Methods Demographic and epidemiological data were analyzed to identify factors possibly related with the microbiological profile of the two collection methods. Associations between categorical and continuous variables were analyzed using the chi-square or Fisher's exact test, or Student's t test. p-Value <0.05 were considered significant. Results Thirty endotracheal tubes and tracheal aspirates samples from 27 subjects were analyzed. Only one patient presented the clinical diagnosis of ventilator-associated pneumonia. Overall, 50% of bacteria were Gram-negative bacilli, followed by Gram-positive bacteria in 37%, and fungi in 10%. No statistically significant difference on the distribution of Gram-positive or Gram-negative bacteria (p = 0.996), and fungi (p = 0.985) were observed between the collection methods. Pseudomonas spp. was the most frequent microorganism identified (23.8%), followed by Streptococcus spp. (18.5%), Acinetobacter spp. (15.9%), coagulase-negative staphylococci (11.2%), and Klebsiella spp. (8.6%). Concordant results between methods amounted to 83.3%. Pseudomonas aeruginosa and Acinetobacter baumannii showed carbapenem resistance in 50% and 43.7% of the isolates, respectively. In general, cultures after endotracheal tubes sonication (non-centrifuged sonication fluid and centrifuged sonication fluid) yielded bacteria with higher rates of antimicrobial resistance compared to tracheal aspirates cultures. Additionally, in 12 subjects (40%), we observed discrepancies regarding microbiologic profiles of cultures performed using the collection methods. Conclusions Our study demonstrated that sonication technique can be applied to ET biofilms to identify microorganisms attached to their surface with a great variety of species identified. However, we did not find significant differences in comparison with the traditional tracheal aspirate culture approach.

Microbial investigation of biofilms recovered from endotracheal tubes using sonication in intensive care unit pediatric patients.

OBJECTIVES: To compare cultured microorganisms identified on endotracheal tubes biofilms through sonication technique with traditional tracheal aspirate collected at extubation of pediatric intensive care unit patients. METHODS: Demographic and epidemiological data were analyzed to identify factors possibly related with the microbiological profile of the two collection methods. Associations between categorical and continuous variables were analyzed using the chi-square or Fisher's exact test, or Student's t test. p-Value <0.05 were considered significant. RESULTS: Thirty endotracheal tubes and tracheal aspirates samples from 27 subjects were analyzed. Only one patient presented the clinical diagnosis of ventilator-associated pneumonia. Overall, 50% of bacteria were Gram-negative bacilli, followed by Gram-positive bacteria in 37%, and fungi in 10%. No statistically significant difference on the distribution of Gram-positive or Gram-negative bacteria (p=0.996), and fungi (p=0.985) were observed between the collection methods. Pseudomonas spp. was the most frequent microorganism identified (23.8%), followed by Streptococcus spp. (18.5%), Acinetobacter spp. (15.9%), coagulase-negative staphylococci (11.2%), and Klebsiella spp. (8.6%). Concordant results between methods amounted to 83.3%. Pseudomonas aeruginosa and Acinetobacter baumannii showed carbapenem resistance in 50% and 43.7% of the isolates, respectively. In general, cultures after endotracheal tubes sonication (non-centrifuged sonication fluid and centrifuged sonication fluid) yielded bacteria with higher rates of antimicrobial resistance compared to tracheal aspirates cultures. Additionally, in 12 subjects (40%), we observed discrepancies regarding microbiologic profiles of cultures performed using the collection methods. CONCLUSIONS: Our study demonstrated that sonication technique can be applied to ET biofilms to identify microorganisms attached to their surface with a great variety of species identified. However, we did not find significant differences in comparison with the traditional tracheal aspirate culture approach.

Evaluation of the effectiveness of manual and automated dialyzers reprocessing after multiple reuses.

A cross-sectional study was conducted to evaluate the effectiveness of manual and automated dialyzer reprocessing. Dialyzers were filled with fluid thioglycollate medium from blood and dialysate chambers after being reprocessed and chemically sterilized with 0.2% peracetic acid. They were incubated for 14 days at 35°C ± 2°C, and microbiologic analysis was performed. Microorganisms were identified in 3 of the 11 samples (27.3%) from the blood chambers: Sphingomonas paucimobilis (2/3) and Penicillium spp (1/3) and in 11 of the 11 samples (100%) from the dialysate chambers: S paucimobilis (7/11), Stenotrophomonas maltophilia (4/11), Pseudomonas aeruginosa (3/11), Candida spp (1/11), and Acinetobacter baumannii (1/11). Of the 4 manually reprocessed dialyzers, gram-positive bacillus were identified in 1 sample (25%) from the blood chamber, and Bacillus spp and Burkholderia spp were identified in 1 sample (25%) from the dialysate chamber. The dialyzers reprocessing can pose risks safety because of exposure patient to microorganisms.

Concentration of Sonication Fluid through Centrifugation Is Superior to Membrane Filtration for Microbial Diagnosis of Orthopedic Implant-Associated Infection.

Microbial identification of orthopedic implant-associated infections using sonication fluid (SF) submitted to a concentration step by membrane filtration (SMF) was compared with the standard centrifugation (SC) method. Among 33 retrieved infected implants, sonication identified microorganisms in 26 (78.8%). The sensitivity of SC was higher than that of SMF (78.8% versus 30.3%; P < 0.001).

Microbiological evaluation of the steam sterilization of assembled laparoscopic instruments / Avaliação microbiológica da esterilização a vapor do instrumental laparoscópico montado / Evaluación microbiológica de la esterilización a vapor de instrumental laparoscópico montado

ABSTRACT Objective: assess the safety of steam sterilization of assembled laparoscopic instruments with challenge contamination. Method: a laboratory experimental study, using as test samples trocars and laparoscopic graspers. Geobacillus stearothermophillus ATCC-7953 was used, with a microbial population of 106UFC/Filter paper substrate, removed from the biological indicator. Three of them were introduced into each instrument at the time of assembly, and sterilized at pressurized saturated steam, 134oC for 5 minutes. After sterilization, the instrument was disassembled and each filter paper substrate was inoculated in soybean casein culture and incubated at 56oC for 21 days. In case of absence of growth, they were subjected to heat shock of 80oC, for 20 minutes and re-incubated for 72 hours. Sample size: 185 graspers and 185 trocars, with 95% power. We paired the experiments with comparative negative control groups (5 graspers and 5 trocars with challenge contamination, sterilized disassembled) and positive control (30 filter paper supports, unsterilized), subject to the same incubation procedures. Results: there was no microbial growth in experimental and negative control. The results of the positive control were satisfactory. Conclusion: this study provided strong scientific evidence to support the safety of steam sterilizing of the assembled laparoscopic instrument.

RESUMO Objetivo: avaliar a segurança da esterilização a vapor, do instrumental laparoscópico montado com desafio da contaminação. Método: estudo experimental laboratorial, cujo corpo de prova foram trocarte e pinça laparoscópica. Utilizou-se esporos Geobacillus stearothermophillus ATCC-7953, com população microbiana de 106UFC/suporte de papel filtro, removidos do indicador biológico. Três deles foram introduzidos no interior de cada instrumento, no momento da montagem, sendo esterilizados a vapor saturado sob pressão, 134oC por 5 minutos. Depois da esterilização, o instrumental foi desmontado, e cada suporte de papel filtro foi inoculado em meio de cultura de caseína soja, incubado a 56oC por 21 dias. Não havendo crescimento, foram submetidos a um choque térmico de 80oC, por 20 minutos e reincubados por 72 horas. Tamanho da amostra, 185 pinças e 185 trocartes, com poder de 95%. Os experimentos foram acompanhados dos grupos controle negativo comparativo (5 pinças e 5 trocartes com contaminação desafio, esterilizados desmontados) e positivo (30 suportes de papel filtro, não esterilizados), submetidos aos mesmos procedimentos de incubação. Resultados: não houve nenhum crescimento microbiano nos grupos experimental e controle negativo. Os resultados do controle positivo foram satisfatórios. Conclusão: este estudo forneceu fortes evidências científicas para sustentar a segurança da prática de esterilização a vapor do instrumental laparoscópico montado.

RESUMEN Objetivo: evaluar la seguridad de la esterilización a través de vapor, de instrumental laparoscópico previamente montado con desafío de contaminación. Método: estudio experimental en laboratorio, cuyo cuerpo de prueba fueron trócarte y pinza laparoscópica. Se utilizó esporas Geobacillus stearothermophilus ATCC-7953, con población microbiana de 106UFC/soporte de papel filtro, removidos del indicador biológico. Tres de ellos fueron introducidos en el interior de cada instrumento, en el momento del montaje, los que fueron esterilizados a vapor saturado bajo presión, 134oC por 5 minutos. Después de la esterilización, el instrumental fue desmontado y cada soporte de papel filtro fue inoculado en medio de una cultura de caseína y soya, incubado a 56oC por 21 días. No habiendo crecimiento, fueron sometidos a un choque térmico de 80oC, por 20 minutos y nuevamente incubados por 72 horas. La muestra estuvo constituida por 185 pinzas y 185 trócartes, con poder de 95%. Los experimentos fueron acompañados en los grupos: control negativo comparativo (5 pinzas y 5 trócartes con contaminación desafío, esterilizados desmontados) y positivo (30 soportes de papel filtro, no esterilizados), sometidos a los mismos procedimientos de incubación. Resultados: no se encontró crecimiento microbiano en los grupos experimental y control negativo. Los resultados del control positivo fueron satisfactorios. Conclusión: este estudio suministra fuertes evidencias científicas para sustentar que la práctica, de esterilización a vapor del instrumental laparoscópico montado, es segura.

Microbial diagnosis of infection and colonization of cardiac implantable electronic devices by use of sonication.

OBJECTIVES: The clinical utility of sonication as an adjunctive diagnostic tool for the microbial diagnosis of cardiac implantable device-associated infections (CIDAIs) was investigated. METHODS: The implants of 83 subjects were investigated, 15 with a CIDAI and 68 without a clinical infection. Clinical data were analyzed prospectively and sonication fluid cultures (83 patients, 100%) and traditional cultures (31 patients, 37.4%) were performed RESULTS: Generator pocket infection and device-related endocarditis were found in 13 (86.7%) and four (26.7%) subjects, respectively. The mean numbers of previous technical complications and infections were higher in the infected patients compared to the non-infected patients (8 vs. 1, p<0.001; 2 vs. 0, p<0.031, respectively). The sensitivity and specificity for detecting CIDAI was 73.3% (11/15) and 48.5% (33/68) for sonication fluid culture, and 26.7% (4/15) and 100% (16/16) for traditional culture (p<0.001), respectively. A higher number of organisms were identified by sonication fluid than by tissue culture (58 vs. 4 specimens; p<0.001). The most frequent organisms cultured were Gram-positive cocci (66.1%), mainly coagulase-negative staphylococci (35.5%). Thirty-five (51.5%) non-infected subjects were considered colonized due to the positive identification of organisms exclusively through sonication fluid culture. CONCLUSIONS: Sonication fluid culture from the removed cardiac implants has the potential to improve the microbiological diagnosis of CIDAIs.

Evaluation of a new rapid test for carbapenemase detection in carbapenem resistant Enterobacteriaceae.

We evaluated a new phenotypic test for carbapenemase detection. A total of 100 Enterobacteriaceae isolates were selected. The test was compared with conventional PCR for bla(KPC) and bla(NDM) detection. We found 100% sensitivity and specificity, suggesting that this test may be a feasible alternative for rapid carbapenemase detection.

Post-photorefractive keratectomy contact lens microbiological findings of individuals who work in a hospital environment.

OBJECTIVE: To describe the microbiological findings from bandage contact lenses in patients who work in a hospital environment submitted to photorefractive keratectomy (PRK). METHODS: This prospective comparative case series enrolled 43 eyes of 22 volunteers (28.05 ± 3.50 years). Fourteen individuals (n = 27) were health care professionals who work in health care facilities or community physician's offices. Eight individuals (n = 16) were patients who do not work in hospital environment. Photorefractive keratectomy was performed using standard technique, and a silicone hydrogel bandage contact lens was placed on the cornea and evaluated for adequate fit. Seven days after surgery, the bandage lenses were removed and imprinted in the following culture media: blood agar, chocolate agar, anaerobic-selective agar, and Sabouraud agar. When microbial growth was detected, the microorganism was identified, colony-forming units were quantified, and morphology and Gram-staining properties were analyzed. All isolates were tested for susceptibility to various antibiotics. Significance was assessed by Fisher exact test. RESULTS: Microbial growth was detected in 16.27% of all contact lenses samples. No fungi or anaerobes were found. Microbial growth was only observed in bandage lenses removed from patients who work in hospital environments. Most microorganisms found were sensitive to all antibiotics tested. CONCLUSION: These results suggest that working in hospital environments increase contamination of the contact lenses after PRK.

Improved diagnosis of infection associated with osteosynthesis by use of sonication of fracture fixation implants.

Previous studies have shown that sonication fluid cultures from removed orthopedic devices improved the microbiological diagnosis of orthopedic implant-associated infections; however, few of these investigations have applied sonication to the removed fracture fixation devices to evaluate its utility for the diagnosis of osteosynthesis-associated infection (OAI). We compared sonication fluid to conventional tissue cultures from 180 subjects with different sizes of plates and screws (n = 156), spinal implants (n = 26), and intramedullary nails (n = 3), of whom 125 and 55 subjects had OAI and noninfected osteosynthesis (NIO), respectively. The sensitivity for detecting OAI was 90.4% for sonication fluid culture and 56.8% for periprosthetic tissue cultures (P < 0.05), and the specificities were 90.9% and 96.4%, respectively. Sonication fluid culture detected more pathogens than peri-implant tissue culture (113 versus 71; P < 0.001), while polymicrobial infections were diagnosed by sonication fluid cultures and tissue cultures in 20.8% and 8% (P < 0.001), respectively. Microbiological diagnosis was achieved exclusively by sonication fluid cultures for 47 (90.4%) subjects, and among them, 18 (38.3%) had previously received antibiotics, whereas in five (9.6%) infected subjects, tissue culture was positive and the sonication fluid culture was negative. Among 39 (31.2%) OAI cases receiving antibiotics, the identification of the organisms occurred in 38.5% and 82.1% of the tissue and sonication fluid cultures, respectively (P < 0.049). We demonstrated that sonication fluid culture from removed osteosyntheses has the potential for improving the microbiological diagnosis of OAI.

Evaluation of silicon oil on bacterial growth.

PURPOSE: To analyze the antimicrobial properties of silicon oil (Óleo de Silicone®, Ophthalmos, Brazil) on in vitro bacterial growth of different microorganisms related to endophthalmitis. METHODS: The following microorganisms were analyzed: (1) Pseudomonas aeruginosa (ATCC 27583); (2) Escherichia coli (ATCC 25922); (3) Staphylococcus aureus (ATCC 25923); (4) Staphylococcus epidermidis (ATCC 12228); (5) Candida albicans (ATCC 10231); (6) Klebsiella pneumoniae (ATCC 13883); and (7) Streptococcus pneumoniae (ATCC 49619). The plates were incubated at 35 ± 2ºC and its growth examined after 24 hours. An empty disk was placed in the center of each plate as a control. RESULTS: No inhibition halos were verified in any of the plates containing the four different concentrations of the bacterial inocula. CONCLUSIONS: The silicon oil 1000 cps does not have any effect on bacterial growth of any of the studied microorganisms.