Evaluation of in vitro activity of ceftolozane-tazobactam against recent clinical bacterial isolates from Brazil - the EM200 study

ABSTRACT Background: The emergence of antibiotic resistance is increasing and there are few effective antibiotics to treat infections caused by resistant and multidrug resistant bacterial pathogens. This study aimed to evaluate the in vitro activity of ceftolozane-tazobactam against clinical bacterial isolates from Brazil. Methods: A total of 673 Gram-negative bacterial isolates including Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa and other Enterobacterales collected from 2016 to 2017 were tested, most of them isolated from patients in intensive care units. Minimum inhibitory concentrations (MIC50/90) were determined by broth microdilution for amikacin, aztreonam, cefepime, cefotaxime, cefoxitin, ceftolozane-tazobactam, ceftazidime, ceftriaxone, ciprofloxacin, colistin, ertapenem, imipenem, levofloxacin, meropenem, and piperacillin-tazobactam using dried panels. Antimicrobial susceptibility results were interpreted according to Clinical and Laboratory Standards Institute criteria. Results: Susceptibility rates to ceftolozane-tazobactam ranged from 40.4% to 94.9%. P. aeruginosa susceptibility rate to ceftolozane-tazobactam was 84.9% (MIC50/90, 1/16 µg/mL) and 99.2% to colistin. For E. coli, ceftolozane-tazobactam inhibited 94.9% (MIC50/90, 0.25/1 µg/mL) of the microorganisms. The susceptibility rate of K. pneumoniae to ceftolozane-tazobactam was 40.4% (MIC50/90, 16/>32 µg/mL). Other Enterobacterales have shown susceptibility rates of 81.1% (MIC50/90, 0.5/16 µg/mL) to ceftolozane-tazobactam, 93.9% to meropenem, 90.9% to amikacin (90.9%), and 88.6% to ertapenem. In non-carbapenemase producing isolates, AmpC mutations were found three isolates. Conclusions: Ceftolozane-tazobactam has shown relevant activity against a large variety of the analyzed microorganisms collected from multiple centers in Brazil, showing promising results even in multidrug resistant strains.

Infections and antimicrobial resistance in an adult intensive care unit in a Brazilian hospital and the influence of drug resistance on the thirty-day mortality among patients with bloodstream infections

Abstract INTRODUCTION: The present study aimed to determine the incidence of health care-associated infections (HCAIs) and identify the main resistant microorganisms in intensive care unit (ICU) patients in a Brazilian university hospital. METHODS: A retrospective cohort study was conducted in a Brazilian teaching hospital between 2012 and 2014. RESULTS: Overall, 81.2% of the infections were acquired in the ICU. The most common resistant pathogenic phenotypes in all-site and bloodstream infections were oxacillin-resistant coagulase-negative staphylococci and carbapenem-resistant Acinetobacter spp. (89.9% and 87.4%; 80.6% and 70.0%), respectively. CONCLUSIONS: There is an urgent need to focus on HCAIs in ICUs in Brazil.

Infecciones en pie diabético: Elección del tratamiento antibiótico empírico / Infections in diabetic foot. Choice of empirical antibiotic regimen

Las infecciones del pie diabético se asocian a complicaciones graves y constituyen la principal causa de hospitalización relacionada con diabetes y amputación de miembros inferiores. Para evitar su progresión, se requiere una conducta inicial rápida y adecuada que incluye toma de muestras para cultivos e inicio inmediato de tratamiento antibiótico empírico, según las características de las lesiones y la prevalencia local de microorganismos. Por ello, es necesario conocer y vigilar la microbiología local y la resistencia a los antimicrobianos. El objetivo de este trabajo fue describir la frecuencia de gérmenes en infecciones de pie diabético en pacientes ambulatorios asistidos en nuestro hospital en 2018 e identificar el esquema antibiótico con mayor cobertura, en comparación con los resultados de un estudio similar realizado en 2015. Fueron analizadas 72 muestras tomadas mediante punción por piel sana de partes blandas. Entre los 68 gérmenes aislados, los Gram negativos fueron los más frecuentes (47.1%), lo que representa un aumento significativo en relación a la frecuencia observada en 2015 (24.6%) p = 0.01 y un aumento de la sensibilidad a ciprofloxacina de 25% a 62.5% (p=0.03). El esquema con mayor cobertura fue amoxicilina-clavulánico con ciprofloxacina (77.9%) mientras que en 2015 fue amoxicilina-clavulánico con trimetoprima sulfametoxazol. La vigilancia de la microbiología local es fundamental para la elección del antibiótico empírico en las infecciones de pie diabético. En nuestro hospital, cuando la infección es de partes blandas, se recomienda la combinación amoxicilina-clavulánico más ciprofloxacina como esquema antibiótico empírico según los hallazgos de este estudio.

Diabetic foot infections are related to severe complications and constitute the main reason for diabetes-related hospitalization and lower limb amputations. A diabetic foot infection requires prompt actions to avoid progression of the infected wound; a soft tissue sample has to be taken for microbiological culture and empiric antibiotic therapy must be started immediately. Empiric antibiotic schemes should be chosen based on the severity of the infection and the local prevalence of microbial causal agents. Therefore, it is important to monitor these indicators. The aim of this study was to determine which microorganisms were more prevalent in cultures of diabetic foot infections during 2018 and what antibiotic combination was better to cover local microbiology, compared with data available from 2015 for a similar cohort. A total of 68 positive cultures were obtained of 72 soft tissue specimens analyzed. The most frequent microorganisms were Gram negative (47.1%), and resulted significantly more frequent than in 2015 (24.6%) p = 0.01. These Gram negative germs also resulted more sensitive to ciprofloxacin than in 2015 (62.5% vs. 25.0%) p = 0.03. Amoxicillin-clavulanate plus ciprofloxacin was the optimal combination therapy in 2018, while in 2015 it was amoxicillin-clavulanate plus trimethoprim sulfamethoxazole. In agreement with these results, we recommend amoxicillin-clavulanate plus ciprofloxacin as the empiric antibiotic regimen of choice for soft tissue infections in diabetic foot. We consider surveillance of local microbiology to be an important tool in the management of diabetic foot infections.

Direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry and real-time PCR in a combined protocol for diagnosis of bloodstream infections: a turnaround time approach

ABSTRACT Bloodstream infections (BSIs) are serious infections associated with high rates of morbidity and mortality. Every hour delay in initiation of an effective antibiotic increases mortality due to sepsis by 7%. Turnaround time (TAT) for conventional blood cultures takes 48 h, forcing physicians to streamline therapy by exposing patients to broad-spectrum antimicrobials. Our objective was (1) to evaluate the accuracy and TAT of an optimized workflow combining direct matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and in-house real-time polymerase chain reaction (PCR) for bacterial identification and antimicrobial resistance profiling directly from positive blood bottles for diagnosing bloodstream infections and (2) to verify the effect of reporting results to medical staff. A total of 103 BSI episodes from 91 patients admitted to three hospitals in São Paulo, Brazil were included. TAT from molecular versus conventional methods was measured and compared. Our protocol showed an overall agreement of 93.5% for genus and 78.5% for species identification; 74.2% for methicillin resistance detection, 89.2% for extended-spectrum β-lactamase profiling, 77.8% for metallo-β-lactamase profiling, and 100% for carbapenemase profile and vancomycin-resistance detection when compared with conventional testing. TAT of molecular sample processing according to our protocol was 38 h shorter than conventional methods. Antimicrobial interventions were possible in 27 BSI episodes. Antimicrobial discontinuation was achieved in 12 BSI episodes while escalation of therapy occurred in 15 episodes. Antimicrobial therapy was inadequate in three (12%) BSI episodes diagnosed using results of molecular testing. Our in-house rapid protocol for identifying both bacteria and antimicrobial resistance provided rapid and accurate results, having good agreement with conventional testing results. These results could contribute to faster antimicrobial therapy interventions in BSI episodes.

Microbiological analysis of contact lens cases: impact of the hospital environment / Análise microbiológica de estojos de lentes de contato: impacto do ambiente nosocomial

ABSTRACT Purpose: The aim of this study was to analyze the bacterial and fungal microbiota found in contact lens cases among two groups of patients to correlate the data on the habits of contact lens users and to evaluate whether there is a difference in the culture results between users of ridged and nonridged contact lens cases. Methods: Two groups (35 patients per group) were included, consisting of hospital workers and those who had not visited a hospital in the past 30 days, and a questionnaire regarding epidemiological data and habits related to contact lens and lens case use was administered. In addition, 140 samples collected from the right and left compartments of each lens case by swabbing the bottom of the wells were tested using bacterioscopy as well as fungal and bacterial cultures via computerized identification of the species. Results: No fungal growth was identified in any of the 70 contact lens cases; however, bacteria were found in 39 cases, and there was no statistical difference between the groups. Most bacteria (>85%) were gram-negative bacilli. Contamination inone compartment of the contact lens case elevated the risk of contamination of the other side (>80%). Moreover, contamination was statistically higher in the ridged cases than in nonridged cases (p=0.0149). Conclusion: The types of bacteria contaminating the cases are generally not seen in eye diseases associated with contact lens use, suggesting that other decisive variables are involved in eye infection from a contaminated lens or case. Fungal contamination of contact lens cases appears to be an exception. Ridged cases are commonly used by contact lens wearers and present a potential risk to eye health. In addition, the results of bacterial tests between hospital workers and those who did not visit a hospital were not significantly different.

RESUMO Objetivo: O objetivo deste trabalho foi analisar a microbiota bacteriana e fúngica encontrada em estojos de lentes de contato em dois grupos, correlacionar os dados sobre os hábitos de uso de lentes de contato e avaliar se há diferença na positividade das culturas entre os usuários estojos de lentes de contato com ranhuras e sem ranhuras. Métodos: Dois grupos foram formados, trabalhadores do hospital e pessoas que não visitaram o hospital (35 indivíduos por grupo), e um questionário foi aplicado sobre dados epidemiológicos e hábitos relacionados ao uso de lentes de contato e estojos de lentes. Além disso, 140 amostras, coletadas do compartimento direito e esquerdo de cada estojo de lente, esfregando o fundo dos mesmos, foram testadas por bacterioscopia e por culturas de fungos e bactérias, com identificação computadorizada da espécie. Resultados: Não houve crescimento fúngico em nenhum dos 70 estojos de lentes de contato, porém bactérias foram encontradas em 39; não houve diferença estatística entre os grupos. A maioria das bactérias (>85%) eram bacilos gram-negativos. Quando um compartimento estava contaminado, o risco de contaminação do outro compartimento era elevado (>80%). A contaminação foi estatisticamente maior nos estojos com ranhuras (p=0,0149). Conclusão: A contaminação dos estojos parece ocorrer por bactérias que, em geral, não são encontradas em doenças oculares associadas ao uso de lentes de contato, sugerindo que existem outras variáveis decisivas nas infecções oculares de uma lente ou estojo contaminado. Contaminação de estojos de lentes de contato com fungos parece ser uma exceção. O uso de estojos com ranhuras é uma prática comum e apresenta um risco potencial à saúde ocular. Não foram encontradas diferenças significativas nos resultados dos testes bacterianos entre trabalhadores hospitalares e pessoas que não visitaram o hospital.

Epidemiology of diabetic foot infections in a reference tertiary hospital in India

Abstract Introduction The present study attempts to examine the microbial profile and antibiotic susceptibility of diabetic foot infections in the intensive care unit of a tertiary referral centre for diabetic foot. As part of the study, we also attempted to find the prevalence of blaNDM-like gene among carbapenem-resistant gram negative infections. Methodology A prospective study of 261 patients with diabetic foot infections was performed during the period between January 2014 and June 2014. Results A total of 289 isolates were obtained from 178 tissue samples from 261 patients, 156 (59.7%) males and 105 (40.2%) females, with a mean age of 58 years (-15 years), having diabetic foot infection. No growth was seen in thirty eight (17.6%) tissue samples. Out of the total samples, 44.3% were monomicrobial and 55.7% were polymicrobial. Gram negative pathogens were predominant (58.5%). Seven of the total isolates were fungal; 0.7% showed pure fungal growth and 1.7% were mixed, grown along with some bacteria. The most frequently isolated bacteria were Staphylococcus aureus (26.9%), followed by Pseudomonas aeruginosa (20.9%). Of the 58.5% gram negative pathogens, 16.5% were Enterobacteriaceae resistant to carbapenems. Among these isolates, 4 (25%) were positive for blaNDM-like gene. Among the rest, 18.6% were carbapenem-resistant Pseudomonas, among which 4 (36.3%) were blaNDM. Among the Staphylococci, 23.7% were methicillin-resistant Staphylococcus aureus. Conclusions Our results support the recent view that gram negative organisms, depending on the geographical location, may be predominant in DFIs. There is an increase in multidrug-resistant pathogens, especially carbapenem resistance and this is creeping rapidly. We need to be more judicious while using empiric antibiotics.

Microorganismos aislados de hemocultivos en niños con cáncer y neutropenia febril de alto riesgo en cinco hospitales de Santiago, Chile, período 2012-2015 / Microorganisms isolated from blood cultures in children with cancer and high-risk febrile neutropenia from five hospitals in Santiago, Chile, 2012-2015

Resumen Antecedentes: Los microorganismos aislados de hemocultivos (HC) en pacientes con neutropenia febril (NF) varían en el tiempo, siendo necesaria su vigilancia para orientar una terapia empírica adecuada. Objetivo: Identificar microorganismos aislados de HC y su perfil de resistencia (R) a antimicrobianos en niños con cáncer y NF de alto riesgo. Método: Estudio prospectivo, multicéntrico de episodios de NF de alto riesgo en pacientes bajo 18 años de edad, de cinco hospitales en Santiago de Chile, 2012-2015. Análisis de HC positivos. Resultados: Se analizaron 206 microorganismos en 185 episodios de NF de alto riesgo con HC positivos. Los aislados principales fueron bacilos gramnegativos (BGN) (46,6%) y cocáceas grampositivas (CGP) (45,1%) y los microorganismos más frecuentes Escherichia coli (22,8%), Staphylococcus coagulasa negativa (18,0%) y Klebsiella spp (16,5%). En resistencia (R) a antimicrobianos destaca: E. coli y Klebsiella spp 4,2 y 67,6% R a cefalosporinas de tercera generación (cefotaxima/ceftriaxona) respectivamente, 10,6 y 40,6% R a ciprofloxacina y 2,1 y 26,5% a amikacina, respectivamente. S. coagulasa negativa y S. aureus 86,4% y 22,2% R a oxacilina, Streptococcus grupo viridans 71% R a penicilina. Discusión: Este estudio actualiza la etiología y el perfil de R de microorganismos aislados en HC de niños con cáncer y NF de alto riesgo, herramienta esencial para el adecuado manejo de estos pacientes.

Background: Microorganisms isolated from blood cultures (BC) in patients with febrile neutropenia (NF) vary over time, requiring systematic monitoring to guide appropriate empirical therapy. Aim: To identify microorganisms isolated from BC and their antimicrobial resistance profile in children with cancer and high risk NF. Method: Prospective, multicenter study. The analysis included episodes of high-risk FN with positive BC in children under 18 years of age treated in five hospitals in Santiago, Chile, 2012-2015. Results: A total of 206 microorganisms were analyzed in 185 episodes of high-risk FN. The main isolates were Gram negative bacilli (46.6%) and Gram positive cocci (45.1%) and the most frequent microorganisms were Escherichia coli (22.8%), coagulase negative Staphylococcus (18.0%) and Klebsiella spp. (16.5%). Escherichia coli and Klebsiella spp showed 4.2% and 67.6% resistance to third generation cephalosporins (cefotaxime/ceftriaxone), 10.6% and 40.6% resistance to fluoroquinolones (ciprofloxacin) and 2.1% and 26.5% to amikacin, respectively. Coagulase negative Staphylococcus and Staphylococcus aureus had 86.4% and 22.2% resistance to oxacillin, Streptococcus viridans group had 71% resistance to penicillin. Discussion: This study updates the etiology and resistance profile of microorganisms isolated in BC from children with cancer and high risk FN, an essential tool for the adequate management of these patients.

Genes qnr en Enterobacteriaceae aisladas en un hospital de Venezuela / qnr genes in Enterobacteriaceae isolated from at a hospital in Venezuela

Resumen Introducción: La resistencia de enterobacterias a quinolonas se ha difundido por el mundo, fenómeno presente también en Venezuela. El mecanismo de esta resistencia pudiera estar mediado por genes incluidos en el cromosoma bacteriano o transmitirse en el interior de plásmidos. Objetivo: Evaluar la resistencia a quino-lonas, codificada por genes qnr, presentes en cepas de enterobacterias, aisladas en el Hospital Universitario de Cumaná, Venezuela. Métodos: A las cepas obtenidas se les realizaron pruebas de susceptibilidad antimicrobiana a quinolonas, β-lactámicos y aminoglucósidos. La presencia del gen qnr se determinó por RPC. Las enterobacterias portadoras del gen qnr fueron sometidas al proceso de conjugación bacteriana para comprobar su capacidad de transferencia. A las transconjugantes obtenidas se les realizó pruebas de susceptibilidad antimicrobiana y RPC para comprobar la transferencia de los genes. Resultados: Se encontraron elevados porcentajes de resistencia antimicrobiana a quinolonas y betalactámicos. El 33,6% de las cepas eran portadoras del gen qnrB, y 0,9% del gen qnrA. Se obtuvieron 23 cepas transconjugantes; de éstas, 20 portaban el gen qnrB, no se observó la presencia de qnrA. Discusión: En conclusión, el elevado porcentaje de genes qnr encontrado en las enterobacterias aisladas, y comprobada la presencia de éstos en plásmidos transferibles, complica la aplicación de tratamientos basados en quinolonas y fluoroquinolonas, por lo que es recomendable el uso racional de estos antimicrobianos, y proponer la rotación de la terapia antimicrobiana, a fin de evitar la selección de cepas resistentes.

Background: Enterobacteria resistant to quinolones is increasing worldwide, including Venezuela. The mechanism for this resistance could be due to genes included in the chromosome or in transmissible plasmids. Aim: To evaluate the resistance to quinolones, coded by qnr genes present in enterobacteria species, isolated in the University Hospital of Cumana, Venezuela. Methods: Antimicrobial susceptibility tests to quinolones, beta-lactams and aminoglycosides were carried out to all the isolates. The presence of qnr genes were determined by PCR. The isolates carrying the qnr genes were used for bacterial conjugation tests to determine the presence of transferable plasmids. Antimicrobial susceptibility tests and PCR were carried out in the transconjugants to verify the transfer of the genes. Results: High levels of antimicrobial resistance to quinolones and beta-lactams were found among the isolates. We found that 33.6% of the isolates carry the qnrB gene and 0.9% qnr A gene. Of the 23 transconjugants, 20 showed to have qnrB gene, but none qnrA. Discussion: We concluded that the high frequency of qnr genes found in the enterobacteria isolates and their presence on transferable plasmids, complicate the use of quinolones for the treatment of bacterial infections, thus, a treatment plan should be designed with the rational use and the rotation of different types of antimicrobials, in order to avoid the selection of increasingly resistant strains.

Prevalencia y perfil de susceptibilidad antimicrobiana en bacterias aisladas de úlceras crónicas infectadas en adultos / Prevalence and susceptibility pattern of bacteria isolated from infected chronic wounds in adult patients

Resumen Introducción: Las úlceras crónicas son un problema de salud pública, agravándose por infecciones bacterianas causadas principalmente por agentes resistentes. Objetivo: Estudiar prevalencia y perfil de susceptibilidad en bacterias aisladas de úlceras crónicas en pacientes adultos. Pacientes y Métodos: Pacientes atendidos en la Fundación Instituto Nacional de Heridas entre mayo y julio de 2014, con úlceras crónicas en extremidades inferiores con signos inflamatorios clínicos. Las muestras fueron cultivadas en aerobiosis y anaerobiosis y para la identificación bacteriana se empleó el sistema de galerías API (Biomerieux). La susceptibilidad in vitro se evaluó según el método de Kirby Bauer. Resultados: Se reclutaron 73 pacientes, entre quienes 46 presentaron úlceras infectadas, diagnosticándose 33 úlceras venosas con predominio de infección polimicrobiana y 10 úlceras de pie diabético con predominio de infección monomicrobiana (p ≤ 0,05). Se aislaron 68 cepas de los 46 pacientes con úlcera infectada. Las enterobacterias predominaron en infección monomicrobiana (p ≤ 0,05) y los demás grupos bacterianos fueron levemente más frecuentes en infección polimicrobiana. La especie prevalente fue Staphylococcus aureus (24%) seguida de Pseudomonas aeruginosa (18%). Cincuenta cepas (77%) presentaron resistencia a uno o más antibacterianos. Destacamos resistencia de S. aureus a ciprofloxacina (50%) y cefoxitina (37,5%) identificándose así resistencia a meticilina en la comunidad (SARM-AC), siendo todas sensibles a cotrimoxazol. Las enterobacterias presentaron resistencia a sensibilidad a amikacina (95,5%), P. aeruginosa evidenció resistencia a ciprofloxacina (33,3%) con alta sensibilidad a gentamicina (91,7%) y amikacina (83,3%), mientras Acinetobacter spp presentó resistencia a ciprofloxacina y ceftazidima en 60%, con 100% de sensibilidad a imipenem. Streptococcus β hemolítico presentó 50% de resistencia a clindamicina y penicilina. Conclusión: Estos datos entregan información epidemiológica de infecciones de úlceras crónicas, representando un apoyo al diagnóstico, tratamiento y manejo de esta patología.

Background: Chronic wounds are considered a public health problem that may be complicated by bacterial infections, mainly caused by resistant strains. Aim: To study the bacteria prevalence and antimicrobial susceptibility in samples from adult patients with chronic wounds. Methods: Patients treated at National Institute of Wounds Foundation between May and July 2014, with chronic ulcers in lower extremities with clinical inflammatory signs were recluted. Samples were cultured in aerobic and anaerobic atmosphere and species identification was performed by API (Biomerieux) galleries. The in vitro susceptibility was evaluated according to the Kirby Bauer method. Results: From 73 patients, 46 had infected wounds most of them were venous ulcers (33) with prevalence in polymicrobial infections and 10 with foot-diabetes ulcers with prevalence in monomicrobial infections (p ≤ 0.05). Sixty-eight strains were isolated and Enterobacteriaceae were predominant in monomicrobial infection (p ≤ 0.05) and the other groups were slightly higher in polymicrobial infection. The main species were Staphylococcus aureus (24%) followed by P. aeruginosa (18%). Fifty strains (77%) were resistant or multi-resistance. We emphasize resistance of S. aureus to ciprofloxacin (50%) and cefoxitin (37.5%), thus identifying resistance to methicillin in the community (CA-SAMR), all of which are sensitive to cotrimoxazole. Enterobacteria showed sensitivity to amikacin (95.5%), P. aeruginosa showed resistance to ciprofloxacin (33.3%) with high sensitivity to gentamicin (91.7%) and amikacin (83.3%), while Acinetobacter spp showed resistance to ciprofloxacin and ceftazidime in 60%, with 100% sensitivity to imipenem. 50% Streptococcus β hemolytic showed resistance to clindamycin and penicillin. Conclusion: These data provide epidemiological information on chronic wound infections, representing support for diagnosis, treatment and management of this pathology.

Is it possible to perform bacterial identification and antimicrobial susceptibility testing with a positive blood culture bottle for quick diagnosis of bloodstream infections?

Abstract INTRODUCTION: Bloodstream infections can be fatal, and timely identification of the etiologic agent is important for treatment. METHODOLOGY: An alternative method, consisting of direct identification and susceptibility testing of blood culture bottles using the automated VITEK 2® system, was assessed. RESULTS: All 37 of the Gram-negative bacilli (GNB) identifications and 57.1% of the 28 Gram-positive cocci (GPC) identifications matched those obtained with standard methods. In susceptibility testing, the agreement was greater than 90%. CONCLUSIONS: This alternative methodology may assist in the early identification and susceptibility testing of GNB. Further research is necessary to develop appropriate methods for GPC.

Chemical characterization, antioxidant and antimicrobial activity of propolis obtained from Melipona quadrifasciata quadrifasciata and Tetragonisca angustula stingless bees

In this study, we investigated the chemical composition, and antioxidant and antibacterial properties of ethanolic extracts of propolis (EEP) from Melipona quadrifasciata quadrifasciata and Tetragonisca angustula. Chemical composition of EEP was determined by colorimetry and chromatographic (HPLC-DAD and UPLC-Q/TOF-MS/MS) analysis. Antimicrobial activity of EEP was evaluated against gram-positive (S. aureus, methicillin-resistant S. aureus, E. faecalis) and gram-negative (E. coli and K. pneumoniae) bacteria by the minimal inhibitory concentration (MIC) test using the microdilution method. Furthermore, the growth curve and integrity of cell membrane of S. aureus and E. coli were investigated using standard microbiological methods. HPLC-DAD analysis showed that the EEP of M. quadrifasciata quadrifasciata has a more complex chemical composition than the EEP of T. angustula. Moreover, UPLC-MS analyses of M. quadrifasciata quadrifascita indicated flavonoids and terpenes as major constituents. The bactericidal activity of both EEPs was higher against gram-positive bacteria than for gram-negative bacteria. The EEP from M. quadrifasciata quadrifasciata presented MIC values lower than the EEP from T. angustula for all tested bacteria. The EEP from M. quadrifasciata quadrifasciata caused lysis of the bacterial wall and release of intracellular components from both E. coli and S. aureus. Our findings indicate that the chemical composition of propolis from stingless bees is complex and depends on the species. The extract from M. quadrifasciata quadrifascita was more effective against gram-positive than gram-negative strains, especially against S. aureus and methicillin-resistant S. aureus compared to T. angustula extract, by a mechanism that involves disturbance of the bacterial cell membrane integrity.

Tissue reaction to Aroeira (Myracrodruon urundeuva) extracts associated with microorganisms: an in vivo study

Abstract: Based on aroeira's (Myracrodruon urundeuva) antimicrobial activity and a future trend to compose intracanal medication, the aim of this study was to assess in vivo inflamatory tissue response to the extracts by edemogenic and histological analysis containing inactivated facultative and anaerobic microorganisms. For edema quantification, eighteen animals were divided into three groups (n = 3, periods: 3 and 6 hours) and 0.2 mL of 1% Evans blue per 100 g of body weight was injected into the penile vein under general anesthesia. After 30 min the animals received a subcutaneous injection in the dorsal region of aqueous or ethanolic extract of aroeira or saline (control) containing inactivated bacteria. Samples were collected, immersed in formamide for 72h, and evaluated by spectrophotometry (630 m). For histological analysis, polyethylene tubes with the extracts were implanted in the dorsal of 30 male rats. Analysis of the fibrous capsule and inflammatory infiltrate were performed after 7 and 30 days. The aqueous extract group induced less edema in both postoperative periods compared to the other groups, but the differences were not significant (p > 0.05). Tissue repair was significantly better after 30 days than after 7 days (p < 0.01). The aqueous solution showed less inflammatory response than the ethanolic solution (p < 0.05), with tendency for better results than control after 7 days. After 30 days, the response to both extracts was similar to control. The aqueous and ethanolic aroeira extracts containing inactivated microorganisms showed a trend for better results than saline, even when associated with microorganisms, and facilitated the tissue repair process.

Etiología y resistencia a antimicrobianos de la infección no complicada del tracto urinario / Etiology and antimicrobial resistance of uncomplicated urinary tract infections

La infección urinaria no complicada en mujeres es un motivo frecuente de consulta e indicación de antimicrobianos. El objetivo de este estudio fue definir etiología y resistencia a antimicrobianos en episodios de infección urinaria no complicada. Este estudio prospectivo incluyó mujeres premenopáusicas no embarazadas, con infección urinaria no complicada, que consultaron en un hospital público y tres centros privados de las ciudades de Buenos Aires y La Plata (2011-2013). La edad media de 138 pacientes con infección confirmada por urocultivo fue 28 años. El diagnóstico fue cistitis en 97 (70%) y pielonefritis en 41 (30%). Las frecuencias de los microorganismos aislados fueron: Escherichia coli 97 (70%), Staphylococcus saprophyticus 24 (17%), Proteus spp. 10 (7%), Klebsiella spp. 5 (4%), Enterococcus spp. 1 (0.7%) y Pseudomonas aeruginosa 1 (0.7%). Las frecuencias de resistencia a antimicrobianos fueron: ampicilina-sulbactam 51 (37%), cefalexina 39 (28%), trimetoprima/sulfametoxazol 31 (22%), nitrofurantoína 17 (12%), gentamicina 10 (7%) y ciprofloxacina 7 (5%). La frecuencia de resistencia a ampicilina-sulbactam, trimetoprima/sulfametoxazol y cefalexina es mayor que las previamente publicadas en Argentina, lo que limita su recomendación para el tratamiento empírico. Una mejor comprensión de la etiología y la susceptibilidad antimicrobiana local permite el diseño de pautas más adecuadas para el tratamiento empírico.

Uncomplicated urinary tract infections rank among the most frequent bacterial infections in women in the outpatient setting and represent a major cause of antimicrobial prescription. The aims of this study were to assess frequencies and antimicrobial resistance of current uropathogens causing uncomplicated urinary tract infection. In a prospective multicenter study, patients were recruited in ambulatory settings of four participating hospitals between June 2011 and December 2013. We analyzed 138 patients that met clinical and bacteriological diagnostic criteria. The mean age was 28 years. Cystitis was defined in 70% (n: 97) and pyelonephritis in 30% (n: 41). Frequencies of isolated microorganisms were: Escherichia coli 70% (n: 97), Staphylococcus saprophyticus 17% (n: 24), Proteus spp. 7% (n: 10), Klebsiella spp. 4% (n: 5), Enterococcus spp. and Pseudomonas aeruginosa 1 (0.7%) each. The antimicrobial resistance was: ampicillin-sulbactam 37% (n: 51) cephalexin 28% (n: 39), trimethoprim/sulfamethoxazole 22% (n: 31), nitrofurantoin 12% (n: 17), gentamicin 7% (n: 10) and ciprofloxacin 5% (n: 7). The levels of resistance found for ampicillin-sulbactam, trimethoprim/sulfamethoxazole and cephalexin were higher than those previously reported in Argentina. A better knowledge of the etiology and local antimicrobial susceptibility allows the design of more adequate guidelines for empirical treatment.

Safety and efficacy of a novel drug elores (ceftriaxone + sulbactam + disodium edetate) in the management of multi-drug resistant bacterial infections in tertiary care centers: a post-marketing surveillance study

Abstract Objective: In India, Elores (CSE-1034: ceftriaxone + sulbactam + disodium edetate) was approved as a broad spectrum antibiotic in year 2011 and is used for management of Extended Spectrum Beta Lactamases/Metallo Beta lactamases infections in tertiary care centers. The objective of this study was to investigate the efficacy of this drug in patients with Extended Spectrum Beta Lactamases/Metallo Beta lactamases infections and identify the incidence of adverse events in real clinical settings. Methods: This Post Marketing Surveillance study was conducted at 17 centers across India and included 2500 patients of all age groups suffering from various bacterial infections and treated with Elores (CSE1034). Information regarding demographic, clinical and microbiological parameters, dosage and treatment duration, efficacy and adverse events (AEs) associated with the treatment were recorded. Results: A total of 2500 patients were included in the study and efficacy was evaluated in 2487 patients. In total, 409 AEs were reported in 211 (8.4%) patients. The major AEs reported were vomiting (3.0%), pain at injection site (2.5%), nausea (2.3%), redness at site (1.96%), thrombophlebitis (1.4%). Of total reported AEs, 40 (5.3%) AEs were reported in pediatric, 310 (20.6%) in adult, and 59 (23.6%) in geriatric group. No AE belonging to grade IV or V was reported in any patient. In terms of efficacy, 1977 (79.4%) patients were cured, 501 (20.1%) patients showed clinical improvement and 5 (0.2%) patients were complete failure. The treatment duration varied from 5 to 7 days in different patients depending on the infection type. Conclusion: In this post-marketing surveillance study, CSE-1034 was found to be an effective and safe option against Pip tazo and meropenem in management of patients with multi-drug resistant (MDR) bacterial infections under routine ward settings.

Direct identification from positive blood broth culture by matrix-assisted laser desorption-ionization time-of-flight mass spectrometry (MALDI-TOF MS)

ABSTRACT Bloodstream infections (BSIs) are among the most concerning bacterial infections. They are one of the leading causes of morbidity and mortality, and occur in 30-70% of critical care patients. The prompt identification of the causative microorganism can help choosing the appropriate antimicrobial therapy that will lead to better clinical outcomes. Blood culture is one of the most relevant tests for microbiological diagnosis of bacterial infections. The introduction of the MALDI-TOF microbiological diagnosis significantly decreased the time of identifying microorganisms. However, it depends on the growth on solid culture medium. In this study, 538 bottles of positive blood cultures were evaluated to test the accuracy of an in house modified protocol. The study sample consisted of 198 Gram-negative and 350 Gram-positive bacteria. In all, 460 (83.94%) species were identified based on the direct plate findings. The protocol allowed the identification of 185/198 (93.43%) of the Gram-negative bacteria, including aerobes, anaerobes, and non-fermenters, and 275/350 (78.85%) of the Gram-positive bacteria. The proposed method has the potential to provide accurate results in comparison to the traditional method with the potential to reduce the turnaround time for the results and optimize antimicrobial therapy in BSI.

Resistance among Gram-negative ESKAPE pathogens isolated from hospitalized patients with intra-abdominal and urinary tract infections in Latin American countries: SMART 2013-2015

ABSTRACT Gram-negative ESKAPE pathogens (Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, and Enterobacter species) are important etiologic agents of nosocomial infection that are frequently resistant to broad-spectrum antimicrobial agents. Gram-negative ESKAPE pathogens were collected from hospitalized patients in 11 Latin American countries from 2013 to 2015 as part of the Study for Monitoring Antimicrobial Resistance Trends (SMART) global surveillance program. In total, 2113 isolates from intra-abdominal infections (IAI) and 970 isolates from urinary tract infections (UTI) were tested against antimicrobial agents using standardized CLSI broth microdilution methodology. Of the agents tested, amikacin demonstrated the highest rates of susceptibility (%) for K. pneumoniae (92.2, 92.3), Enterobacter spp. (97.5, 92.1), and P. aeruginosa (85.3, 75.2) isolates from both IAI and UTI, respectively. Ertapenem (68.5, 62.6) and imipenem (79.2, 75.9) showed substantially higher rates of susceptibility (%) than other β-lactams, including piperacillin-tazobactam (35.9, 37.4) against ESBL-positive isolates of K. pneumoniae from IAI and UTI, respectively. Rates of susceptibility to all agents tested against A. baumannii were ≤30.9%. Gram-negative ESKAPE pathogens isolated from Latin America demonstrated compromised in vitro susceptibility to commonly prescribed broad-spectrum, parenteral antimicrobial agents. Continued surveillance is warranted. New antimicrobial agents with potent activity against Gram-negative ESKAPE pathogens are urgently needed.

Phenol degradation and genotypic analysis of dioxygenase genes in bacteria isolated from sediments

Abstract The aerobic degradation of aromatic compounds by bacteria is performed by dioxygenases. To show some characteristic patterns of the dioxygenase genotype and its degradation specificities, twenty-nine gram-negative bacterial cultures were obtained from sediment contaminated with phenolic compounds in Wuhan, China. The isolates were phylogenetically diverse and belonged to 10 genera. All 29 gram-negative bacteria were able to utilize phenol, m-dihydroxybenzene and 2-hydroxybenzoic acid as the sole carbon sources, and members of the three primary genera Pseudomonas, Acinetobacter and Alcaligenes were able to grow in the presence of multiple monoaromatic compounds. PCR and DNA sequence analysis were used to detect dioxygenase genes coding for catechol 1,2-dioxygenase, catechol 2,3-dioxygenase and protocatechuate 3,4-dioxygenase. The results showed that there are 4 genotypes; most strains are either PNP (catechol 1,2-dioxygenase gene is positive, catechol 2,3-dioxygenase gene is negative, protocatechuate 3,4-dioxygenase gene is positive) or PNN (catechol 1,2-dioxygenase gene is positive, catechol 2,3-dioxygenase gene is negative, protocatechuate 3,4-dioxygenase gene is negative). The strains with two dioxygenase genes can usually grow on many more aromatic compounds than strains with one dioxygenase gene. Degradation experiments using a mixed culture representing four bacterial genotypes resulted in the rapid degradation of phenol. Determinations of substrate utilization and phenol degradation revealed their affiliations through dioxygenase genotype data.

Etiología aerobia de apendicitis aguda en adultos: Estudio multicéntrico de la sepsis abdominal en Argentina / Aerobic etiology of acute appendicitis in adults. Multicenter study of abdominal sepsis in Argentina

El tratamiento antibiótico de las apendicitis agudas se decide empíricamente basándose en la información epidemiológica. Las resistencias son variables entre regiones y los datos de Argentina son escasos. En el contexto de un estudio multicéntrico, observacional, de infecciones abdominales, se efectuó el análisis de los pacientes adultos con diagnóstico de apendicitis, incorporados al estudio entre enero 2014 y junio 2015, en 16 centros de 5 provincias argentinas. El objetivo fue analizar los gérmenes aeróbicos prevalentes, su resistencia a antibióticos y el patrón de prescripción antimicrobiana. Se estudiaron 131 apendicitis. Se aislaron 184 bacterias aerobias (1.4 bacterias/episodio): Escherichia coli 106 (57.6%), Klebsiella spp 16 (8.7%), Pseudomonas aeruginosa 19 (10.3%), Enterobacter spp. 2 (1%), otros bacilos Gram negativos 5 (2.7%). Enterococcus spp. 16 (8.7%) y otros cocos Gram positivos 20 (10.9%). La resistencia de E. coli y enterobacterias a ampicilina/sulbactam fue mayor a 34% y a ciprofloxacina mayor a 31%. En cambio, la resistencia de enterobacterias a piperacilina/tazobactam fue 4.8%, a ceftriaxona 9.5% y no se halló resistencia a carbapenemes. Respecto a amikacina fue 3.6% y a gentamicina 8.2%. En función de los resultados, el uso de quinolonas o de ampicilina/sulbactam para el tratamiento de las apendicitis debiera ser desaconsejado. Los esquemas basados en aminoglucósidos debieran ser jerarquizados en función de la sensibilidad hallada y su bajo impacto en la inducción de resistencias.

Antibiotic treatment for acute appendicitis is empirically chosen, based on epidemiological information. Resistance rates are different between regions and there are limited data on the situation in Argentina. As a part of a multicenter, observational study of abdominal infections, we performed the analysis of adult patients diagnosed with appendicitis, enrolled in 16 centers of 5 provinces, between Jan/01/2014 and Jun/30/2015. The aim was to analyze the prevalent aerobic pathogens, their resistance rates and the antimicrobial prescription pattern. On a total of 131 appendicitis cases analyzed, we found 184 aerobic pathogens (1.4 bacteria/episode): Escherichia coli 106 (57.6%), Klebsiella spp 16 (8.7%), Pseudomonas aeruginosa 19 (10.3%), Enterobacter spp. 2 (1%), other Gram negative bacilli 5 (2.7%); Enterococcus spp. 16 (8.7%) and other Gram positive cocci 20 (10.9%). The resistance rate of E. coli and enterobacteria to ampicillin/sulbactam was greater than 34% and greater than 31% to ciprofloxacin. However, the resistance of enterobacteria to piperacillin/tazobactam was 4.8%, to ceftriaxone 9.5%, to amikacin 3.6% and 8.2% to gentamicin. No resistance to carbapenems was found. The choice of quinolones or ampicillin/sulbactam for the treatment of appendicitis should be discouraged in our context, due to the high rates of resistance found in this prevalent etiology. Aminoglycoside-based treatments should be considered, given the findings of high antibiotic susceptibility and their low impact on the induction of resistance.

Evaluación de la susceptibilidad directa desde hemocultivos positivos utilizando el sistema Vitek 2: comparación de dos protocolos rápidos / Evaluation of antimicrobial susceptibility testing directly from positive blood cultures using the Vitek 2 system: comparison of two rapid protocols

Antimicrobial susceptibility testing using Vitek® 2 (bioMérieux) cards inoculated directly from positive blood cultures was compared with the standard method for gramnegative rods. Two different protocols for the inoculum preparation were evaluated. Protocol 2 was faster and easier to perform. Rapid susceptibility testing was accurate and reduced time to results in 24 h.

Se comparó el ensayo de susceptibilidad a los antimicrobianos utilizando las tarjetas Vitek® 2 (bioMérieux) inoculadas directamente de hemocultivos positivos con el método estándar para bacilos gramnegativos. Se evaluaron dos protocolos diferentes para la preparación del inóculo. El segundo protocolo fue más rápido y fácil de realizar. Las pruebas de susceptibilidad rápida fueron precisas y redujeron el tiempo de los resultados en 24 h.

Changes in the conjunctival bacterial flora of patients hospitalized in an intensive care unit / Mudanças na flora bacteriana conjuntival de pacientes internados em unidade de terapia intensiva

ABSTRACT Purpose: To identify the changes in aerobic conjunctival bacterial flora and to correlate culture results with physical health and the duration of patients' hospitalization in an intensive care unit (ICU). Methods: Patients hospitalized in the ICU were included in this study. Conjunctival cultures from all patients were obtained using a standard technique on days 1, 3, 7, and 14. Swabs were plated on nonselective (blood agar) and enriched (chocolate agar) media within one hour. Visible colonies were isolated, and standard microbiological techniques were used to identify the bacteria. The frequency, identity, and correlation of culture results with patients' physical findings and the duration of hospitalization were determined. Results: We obtained 478 cultures (day 1, 270; day 3, 156; day 7, 36; and day 14, 16) from 135 patients; 288 (60.2%) cultures were positive, and 331 microorganisms were isolated. The most frequently isolated microorganism from the cultures was coagulase-negative Staphylococcus species (n=210/331, 63.5%), and the others were Corynebacterium diphtheriae (n=52/331, 15.7%), S. aureus (n=26/331, 7.9%), gram-negative bacilli other than Pseudomonas (n=14/331, 4.2%), Neisseria species (n=8/331, 2.4%), Pseudomonas aeruginosa (n=6/331, 1.8%), Haemophilus influenzae (n=7/331, 2.1%), Acinetobacter species (n=6/331, 1.8%), and Streptococcus species (n=2/331, 0.6%). The frequency of positive cultures significantly increased (p<0.03) with time. Conclusions: Prolonged hospitalization significantly predisposes to bacterial colonization. The colonization rate of S. aureus and Neisseria spp. increased significantly after one week.

RESUMO Objetivo: Identificar as mudanças na flora bacteriana aeróbia da conjuntiva e correlacionar os resultados da cultura com o estado de saúde física e a duração da hospitalização em pacientes em uma unidade de terapia intensiva (UTI). Método: Pacientes que estavam na UTI foram incluídos neste estudo. Culturas conjuntivais foram obtidas nos dias 1, 3, 7 e 14 de todos os pacientes com uma técnica normalizada. Zaragatoas foram semeadas em placas não seletivas (ágar sangue) e enriquecidas (ágar chocolate) dentro de uma hora. Colônias visíveis foram separadas, isoladas, e identificadas utilizando técnicas microbiológicas convencionais. A frequência, identificação e correlação da cultura resulta com achados físicos e a duração da hospitalização foram determinados. Resultados: Um total de 478 culturas (no primeiro dia 270, terceiro dia 156, sétimo dia 36 e dia catorze 16 culturas) foram obtidas de 135 pacientes hospitalizados durante o estudo. Duzentos e oitenta e oito (60,2% de todas as culturas obtidas) culturas foram positivas. Trezentos e trinta e um microrganismos foram isolados a partir dessas culturas. Em todos os grupos, o microrganismo mais frequentemente isolado foi o Staphylococcus species coagulase negativo (n=210/331, 63,5% de todos os microrganismos isolados). Outras bactérias isoladas foram Corynebacterium diphteriae (n=52/331, 15,7%), Staphylococcus aureus (n=26/331, 7,9%), bacilos Gram-negativos que não sejam Pseudomonas (n=14/331, 4,2%), Neisseria species (n=8/331, 2,4%), Pseudomonas aeruginosa (n=6/331, 1,8%), Haemophilus influenzae (n=7/331, 2,1%), Acinetobacter species (n=6/331, 1,8%), e Streptococcus species (n=2/331, 0,6%). Como o tempo de hospitalização prolongada, a positividade em culturas aumentou significativamente (p<0,03). Conclusões: hospitalização prolongada predispõe significativamente a frequência de colonização bacteriana. A taxa de colonização de S. aureus e Neisseria spp. aumentou significativamente depois de uma semana.