Microbiological environmental contamination in a Pediatric Intensive Care Unit / Contaminação ambiental microbiológica em Unidade de Terapia Intensiva Pediátrica / Contaminación ambiental microbiológica en una Unidad de Cuidados Intensivos Pediátricos

Background and objectives: inanimate surfaces and equipment in the hospital environment are considered reservoirs of resistant and pathogenic microorganisms. In Pediatric Intensive Care Units, the risk of infection is also related to the severity of pathologies associated with the immaturity of the immune system of this population. This study aimed to investigate microbiological environmental contamination in a Pediatric Intensive Care Unit. Method: this is an exploratory cross-sectional study, carried out in a Pediatric Intensive Care Unit of a highly complex university hospital, located in southern Brazil. To assess environmental contamination, sterile swabs were rubbed on surfaces corresponding to the patient unit and in the common area. Results: twenty-eight surfaces were analyzed, 12 of which were located in units occupied by patients at the time of collection and 16 surfaces in the common use area. In the total number of surfaces analyzed by microbiological cultures, the patient unit showed 66.67% contamination by microorganisms, while surfaces in the common area showed 56.25%. Regarding the microbiological profile, all isolated microorganisms were Gram-positive and showed resistance, namely Staphylococcus aureus and coagulase-negative Staphylococcus. Conclusion: there was evidence of a high frequency of contamination on inanimate surfaces and equipment near and far from patients, essentially by pathogenic and multi-resistant microorganisms to antimicrobials.(AU)

Justificativa e objetivos: superfícies e equipamentos inanimados no ambiente hospitalar são considerados reservatórios de microrganismos resistentes e patogênicos. Nas Unidades de Cuidados Intensivos Pediátricos, o risco de infeção também está relacionado com a gravidade das patologias associadas à imaturidade do sistema imunitário desta população. Este estudo teve como objetivo investigar a contaminação microbiológica ambiental em uma Unidade de Terapia Intensiva Pediátrica. Método: trata-se de um estudo exploratório transversal, realizado em uma Unidade de Terapia Intensiva Pediátrica de um hospital universitário de alta complexidade, localizado no Sul do Brasil. Para avaliar a contaminação ambiental, foram esfregados swabs estéreis nas superfícies correspondentes à unidade do paciente e na área comum. Resultados: foram analisadas vinte e oito superfícies, sendo 12 localizadas em unidades ocupadas por pacientes no momento da coleta e 16 superfícies em área de uso comum. No total de superfícies analisadas por culturas microbiológicas, a unidade paciente apresentou 66,67% de contaminação por microrganismos, enquanto as superfícies da área comum apresentaram 56,25%. Quanto ao perfil microbiológico, todos os microrganismos isolados eram Gram-positivos e apresentavam resistência, nomeadamente Staphylococcus aureus e Staphylococcus coagulase-negativa. Conclusão: houve evidência de elevada frequência de contaminação em superfícies inanimadas e equipamentos próximos e distantes dos pacientes, essencialmente por microrganismos patogênicos e multirresistentes aos antimicrobianos.(AU)

Fundamento y objetivos: las superficies y equipos inanimados del ambiente hospitalario son considerados reservorios de microorganismos resistentes y patógenos. En las Unidades de Cuidados Intensivos Pediátricos el riesgo de infección también se relaciona con la gravedad de patologías asociadas a la inmadurez del sistema inmunológico de esta población. Este estudio tuvo como objetivo investigar la contaminación ambiental microbiológica en una Unidad de Cuidados Intensivos Pediátricos. Método: se trata de un estudio exploratorio transversal, realizado en una Unidad de Cuidados Intensivos Pediátricos de un hospital universitario de alta complejidad, ubicado en el sur de Brasil. Para evaluar la contaminación ambiental se frotaron hisopos estériles en las superficies correspondientes a la unidad de pacientes y en el área común. Resultados: se analizaron veintiocho superficies, 12 de las cuales estaban ubicadas en unidades ocupadas por los pacientes en el momento de la recogida y 16 superficies en el área de uso común. Del total de superficies analizadas por cultivos microbiológicos, la unidad de pacientes presentó un 66,67% de contaminación por microorganismos, mientras que las superficies del área común presentaron un 56,25%. En cuanto al perfil microbiológico, todos los microorganismos aislados fueron Gram positivos y presentaron resistencia, concretamente Staphylococcus aureus y Staphylococcus coagulasa negativo. Conclusión: se evidenció alta frecuencia de contaminación en superficies inanimadas y equipos cercanos y lejanos de los pacientes, esencialmente por microorganismos patógenos y multirresistentes a los antimicrobianos.(AU)

Genotyping Characteristics of Human Fecal Escherichia coli and Their Association with Multidrug Resistance in Miyun District, Beijing / 生物医学与环境科学（英文）

OBJECTIVE@#To explore the genotyping characteristics of human fecal Escherichia coli( E. coli) and the relationships between antibiotic resistance genes (ARGs) and multidrug resistance (MDR) of E. coli in Miyun District, Beijing, an area with high incidence of infectious diarrheal cases but no related data.@*METHODS@#Over a period of 3 years, 94 E. coli strains were isolated from fecal samples collected from Miyun District Hospital, a surveillance hospital of the National Pathogen Identification Network. The antibiotic susceptibility of the isolates was determined by the broth microdilution method. ARGs, multilocus sequence typing (MLST), and polymorphism trees were analyzed using whole-genome sequencing data (WGS).@*RESULTS@#This study revealed that 68.09% of the isolates had MDR, prevalent and distributed in different clades, with a relatively high rate and low pathogenicity. There was no difference in MDR between the diarrheal (49/70) and healthy groups (15/24).@*CONCLUSION@#We developed a random forest (RF) prediction model of TEM.1 + baeR + mphA + mphB + QnrS1 + AAC.3-IId to identify MDR status, highlighting its potential for early resistance identification. The causes of MDR are likely mobile units transmitting the ARGs. In the future, we will continue to strengthen the monitoring of ARGs and MDR, and increase the number of strains to further verify the accuracy of the MDR markers.

The past, present and future of tuberculosis treatment / 浙江大学学报·医学版

Tuberculosis (TB) is an ancient infectious disease. Before the availability of effective drug therapy, it had high morbidity and mortality. In the past 100 years, the discovery of revolutionary anti-TB drugs such as streptomycin, isoniazid, pyrazinamide, ethambutol and rifampicin, along with drug combination treatment, has greatly improved TB control globally. As anti-TB drugs were widely used, multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains of Mycobacterium tuberculosis emerged due to acquired genetic mutations, and this now presents a major problem for effective treatment. Genes associated with drug resistance have been identified, including katG mutations in isoniazid resistance, rpoB mutations in rifampin resistance, pncA mutations in pyrazinamide resistance, and gyrA mutations in quinolone resistance. The major mechanisms of drug resistance include loss of enzyme activity in prodrug activation, drug target alteration, overexpression of drug target, and overexpression of the efflux pump. During the disease process, Mycobacterium tuberculosis may reside in different microenvironments where it is expose to acidic pH, low oxygen, reactive oxygen species and anti-TB drugs, which can facilitate the development of non-replicating persisters and promote bacterial survival. The mechanisms of persister formation may include toxin-antitoxin (TA) modules, DNA protection and repair, protein degradation such as trans-translation, efflux, and altered metabolism. In recent years, the use of new anti-TB drugs, repurposed drugs, and their drug combinations has greatly improved treatment outcomes in patients with both drug-susceptible TB and MDR/XDR-TB. The importance of developing more effective drugs targeting persisters of Mycobacterium tuberculosis is emphasized. In addition, host-directed therapeutics using both conventional drugs and herbal medicines for more effective TB treatment should also be explored. In this article, we review historical aspects of the research on anti-TB drugs and discuss the current understanding and treatments of drug resistant and persistent tuberculosis to inform future therapeutic development.

Antiadhesion and antibiofilm potential of Fagonia indica from Cholistan desert against clinical multidrug resistant bacteria / Potencial de antiadesão e antibiofilme de Fagonia indica do deserto do Cholistão contra bactérias multirresistentes clínicas

High resistance to antimicrobials is associated with biofilm formation responsible for infectious microbes to withstand severe conditions. Therefore, new alternatives are necessary as biofilm inhibitors to control infections. In this study, the antimicrobial and antibiofilm activities of Fagonia indica extracts were evaluated against MDR clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica has antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against multidrug resistant (MDR) clinical isolates. The extract exhibited its antibiofilm effect by altering adherence and disintegration of bacterial cell wall. Fagonia indica had antibacterial effect as minimum inhibitory concentration (MIC) values ranging from 125 to 500 µg mL-1 and minimum bactericidal concentration (MBC) value was 500-3000 µg mL-1 against MDR isolates. The maximum inhibitory effects of Fagonia indica chloroform extract on biofilm formation was observed on Staphylococcus aureus (71.84%) followed by Klebsiella pneumoniae (70.83%) after 48 hrs showing that inhibition is also time dependent. Our results about bacterial cell protein leakage indicated that MDR isolates treated with chloroform extract of Fagonia indica showed maximum protein leakage of K. pneumoniae (59.14 µg mL-1) followed by S. aureus (56.7 µg mL-1). Cell attachment assays indicated that chloroform extract resulted in a 43.5-53.5% inhibition of cell adherence to a polystyrene surface. Our results revealed that extracts of Fagonia indica significantly inhibited biofilm formation among MDR clinical isolates, therefore, could be applied as antimicrobial agents and cost effective biofilm inhibitor against these MDR isolates.

A alta resistência aos antimicrobianos está associada à formação de biofilme responsável por micróbios infecciosos para suportar condições severas. Portanto, novas alternativas são necessárias como inibidores de biofilme para controlar infecções. Neste estudo, as atividades antimicrobiana e antibiofilme dos extratos de Fagonia indica foram avaliadas contra isolados clínicos MDR. O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica tem efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-1, e valor de concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados clínicos multirresistentes (MDR). O extrato exibiu seu efeito antibiofilme ao alterar a aderência e a desintegração da parede celular bacteriana. Fagonia indica teve efeito antibacteriano com valores de concentração inibitória mínima (CIM) variando de 125 a 500 µg mL-1, e concentração bactericida mínima (MBC) de 500-3000 µg mL-1 contra isolados MDR. Os efeitos inibitórios máximos do extrato de clorofórmio Fagonia indica na formação de biofilme foi observada em Staphylococcus aureus (71,84%), seguido por Klebsiella pneumoniae (70,83%) após 48 horas, mostrando que a inibição também é dependente do tempo. Nossos resultados sobre extravasamento de proteínas de células bacterianas indicaram que isolados MDR tratados com extrato clorofórmico de Fagonia indica apresentaram vazamento máximo de proteínas de K. pneumoniae (59,14 µg mL-1), seguido por S. aureus (56,7 µg mL-1). Ensaios de fixação de células indicaram que o extrato de clorofórmio resultou em uma inibição de 43,5-53,5% da aderência das células a uma superfície de poliestireno. Nossos resultados revelaram que extratos de Fagonia indica inibiram significativamente a formação de biofilme entre isolados clínicos MDR, portanto, poderiam ser aplicados como agentes antimicrobianos e inibidores de biofilme de baixo custo contra esses isolados MDR.

Isolation and identification of multidrug-resistant Escherichia coli from cattle, sheep, poultry and human in Cumilla, Bangladesh

Aims@#Antimicrobial resistance (AMR) is a significant public health concern of modern civilization. The potential risk of AMR is significant in terms of both human and animal health. This study aims to assess the antimicrobial resistance pattern of selected antimicrobials against Escherichia coli of animal, poultry and human origin in the Cumilla district of Bangladesh.@*Methodology and results@#A total of 200 samples were collected from different sources. Isolation and identification of commensal E. coli were performed following standard bacteriological and molecular techniques. Antimicrobial susceptibility testing was performed following the Kirby-Bauer disc diffusion technique. Ampicillin, tetracycline and sulfamethoxazole-trimethoprim resistance genes were detected by polymerase chain reactions (PCR). A total of 152 (76%; 95% confidence interval (CI) 70-81%) E. coli were isolated from cattle, sheep, chicken and human, where 37.5% of isolates were found to be multidrug-resistant (MDR). In the cultural sensitivity test, E. coli showed the highest resistance to sulfamethoxazole-trimethoprim (71%), tetracycline (63%), ampicillin (62%), where gentamicin (23%) showed the lowest resistance, followed by ceftriaxone (26%). The prevalence of resistance genes like blaTEM, tetA, tetB, tetC, sul1 and sul2 were 100%, 95%, 11%, 8%, 58% and 52%, respectively.@*Conclusion, significance and impact of study@#The emergence of multidrug-resistant commensal E. coli and resistance genes circulating in animals, poultry and humans limit the treatment options for serious infections.

Actualización en fibrosis quística - fagoterapia: ¿es el futuro para tratar bacterias multirresistentes? / Phage therapy: is the future to treat multiresistant bacteria?

Las infecciones respiratorias representan una morbilidad y mortalidad significativas, con aumento progresivo de la resistencia a los antibióticos. La escasez de nuevos antibióticos disponibles y la pérdida de eficacia de los antiguos, ha impulsado a investigar otras alternativas de tratamiento. La terapia con bacteriófagos (fagos) representa uno de esos enfoques, la que ha demostrado ser eficaz contra una variedad de patógenos bacterianos, incluidas las cepas resistentes a los medicamentos. La administración puede ser tópica, intravenosa o inhalada, esta última requiere preparaciones estables de fagos y sistemas adecuados para proporcionar partículas que accedan al árbol respiratorio. En esta comunicación se revisan diversos aspectos de los bacteriófagos, los que podrían ser de gran utilidad para el tratamiento de las infecciones pulmonares en pacientes con diagnóstico de fibrosis quística.

Respiratory infections represent a significant morbidity and mortality, with a progressive increase in resistance to antibiotics. The scarcity of new antibiotics available and the loss of efficacy of the old ones has prompted investigation of other treatment alternatives. Bacteriophage (phage) therapy represents one such approach that has been shown to be effective against a variety of bacterial pathogens, including resistant strains to medications. Administration can be topical. Intravenous or inhaled, the latter requiring stable preparations of phages and adequate systems to provide particles that will access the respiratory tree. In this communication various aspects of bacteriophages and their clinical utility are reviewed, which could be very useful for the treatment of pulmonary infections in patients diagnosed with cystic fibrosis.

Resistencia a fármacos antituberculosos en Cuba, 2015-2017 / Resistance to tuberculosis drugs in Cuba, 2015-2017

Introducción: La tuberculosis persiste como un importante problema de salud mundial. En el 2016 se estimaron 600 000 casos de resistente a rifampicina, y entre estos 490 000 casos multidrogorresistentes. Objetivo: Describir el comportamiento de la resistencia de los aislados de M. tuberculosis de pacientes con tuberculosis pulmonar notificados en Cuba entre los años 2015-2017. Métodos: Se determinó la susceptibilidad a isoniacida y rifampicina mediante el método de la nitratasa. A los aislados resistentes a rifampicina/multidrogorresistentes se les determinó mediante el método proporcional la susceptibilidad a ofloxacina, kanamicina, amikacina y capreomicina. Resultados: El 93,2 por ciento de los aislados fueron sensibles a isoniacida y rifampicina. En 39 se identificó resistencia a isoniacida y 23 fueron resistente a rifampicina. Se identificaron 10 multidrogorresistentes. No se detectó resistencia a fármacos de segunda línea. Conclusiones: Los resultados alertan sobre la necesidad de investigar las causas que han conllevado al incremento de la tuberculosis resistente en Cuba(AU)

Introduction: Tuberculosis continues to be an important health problem worldwide. In the year 2016, as many as 600 000 cases of rifampicin resistance were estimated, among which 490 000 were multi-drug resistant. Objective: Describe the behavior of resistance to M. tuberculosis isolates in patients with pulmonary tuberculosis reported in Cuba in the period 2015-2017. Methods: Susceptibility to isoniazid and rifampicin was determined by the nitratase method. Susceptibility of rifampicin resistant / multi-drug resistant isolates to ofloxacin, kanamycin, amikacin and capreomycin was determined by the proportional method. Results: Of the isolates analyzed, 93.2 percent were sensitive to isoniazid and rifampicin. Isoniazid resistance was identified in 39 and 23 were rifampicin resistant. Ten multi-drug resistant isolates were identified. Resistance to second line drugs was not detected. Conclusions: Results warn about the need to study the factors leading to the increase in resistant tuberculosis in Cuba(AU)

Caracterización de bacilos gramnegativos multi-resistentes, aislados en pacientes hospitalizados en instituciones de salud de Barranquilla (Colombia) / Characterization of multiresistant gram-negative bacilli, isolated in patients hospitalized in health institutions in Barranquilla (Colombia)

Resumen Introducción: La resistencia a carbapenémicos en bacilos gramnegativos es un problema de salud pública mundial, debido a que se asocia con altas tasas de mortalidad, aumento en los niveles de resistencia a otros antimicrobianos, elevación en el potencial de diseminación e incremento en los costos de atención en salud. Objetivo: Caracterizar bacilos gramnegativos multirresistentes, aislados en pacientes hospitalizados en instituciones de salud de Barranquilla (Colombia). Material y Métodos: Estudio descriptivo acerca de la caracterización fenotípica y genotípica de la resistencia bacteriana en las infecciones asociadas a la atención en salud, mediada por carbapenemasas en aislados bacterianos enviados por los laboratorios pertenecientes a la red de laboratorios del Departamento del Atlántico. Resultados: La KPC fue la carbapenemasa más frecuente en las Enterobacterales (27,6%), predominando en Klebsiella pneumoniae (13,1%) sola y asociada a otras carbapenemasas. En Pseudomonas aeruginosa predominó la carbapenemasa VIM (32,8%) y la OXA en Acinetobacter baumannii (17,1%). Conclusión: Se encontró una amplia distribución de cepas multi-resistentes productoras de carbapenemasas en instituciones de salud de Barranquilla, las cuales expresaron los siguientes mecanismos de resistencia: KPC, VIM, NDM, OXA.

Abstract Background: The emergence of carbapenem resistant gramnegative bacilli has become a problem of public health worldwide, because it is associated with high mortality rates, increased levels of resistance to other antimicrobials, increased potential for dissemination transition and increase in health care costs. Aim: To characterize multiresistant gram-negative bacilli, isolated in patients hospitalized in health institutions of Barranquilla (Colombia). Methods: A descriptive study was conducted on the phenotypic and genotypic characterization of bacterial resistance in infections associated with health care, mediated by carbapenemases in bacterial isolates sent by laboratories belonging to the laboratory network of the Department of Atlántico. Results: KPC was the most frequent carbapenemase in Enterobacterales (27.6%), predominantly in Klebsiella pneumoniae (13.1%) alone and associated with other carbapenemases. In Pseudomonas aeruginosa, VIM carbapenemase (32.8%) predominated and OXA in Acinetobacter baumannii (17.1%). Conclusion: A wide distribution of multi-resistant strains producing carbapenemases in Atlantic health institutions was found, which expressed the following resistance mechanisms: KPC, VIM, NDM, OXA.

Resistencia enzimática en Pseudomonas aeruginosa, aspectos clínicos y de laboratorio / Enzymatic resistance in Pseudomonas aeruginosa, clinical and laboratory aspects

Resumen Pseudomonas aeruginosa es uno de los principales patógenos que causa infecciones asociadas a la atención en salud (IAAS). Su capacidad de adaptación, diseminación, resistencia intrínseca a los antimicrobianos y de adquirir nuevos mecanismos a través de elementos genéticos móviles, hacen que el tratamiento de las infecciones por este microorganismo sea un desafío para el médico clínico. Intrínsecamente, P. aeruginosa, presenta una reducida permeabilidad en la membrana externa, debido a la expresión de bombas de expulsión, y una cefalosporinasa tipo AmpC inducible. Además, P. aeruginosa es capaz de adquirir nuevos determinantes de resistencia por transferencia horizontal en forma de casetes situados en integrones, y a su vez, localizados en transposones o plásmidos. Dentro de la resistencia enzimática que presenta P. aeruginosa destacan las β-lactamasas, incluyendo aquellas de espectro extendido (BLEE) y las carbapenemasas. Pero también enzimas modificadoras de los aminoglucósidos, haciendo que este microorganismo pueda presentar fenotipos de multi-resistencia (MDR), resistencia extrema (XDR) y panresistencia (PDR) a los antimicrobianos denominados antipseudomonas, incluyendo a las nuevas cefalosporinas con inhibidores de beta-lactamasas.

Abstract Pseudomonas aeruginosa is one of the major pathogens causing healthcare-associated infections (HAI). Its capacity of adaptation, dissemination, intrinsic resistance to antimicrobials and of acquiring new mechanisms through mobile genetic elements, make the treatment of infections by this microorganism a challenge for the clinician. Intrinsically, P. aeruginosa, presents a reduced permeability in the external membrane, due to the expression of efflux pumps, and an inducible AmpC-type cephalosporinase. In addition, P. aeruginosa is able to acquire new resistance determinants by horizontal transfer in the form of cassettes located in integrons, and in turn located in transposons or plasmids. Within the enzymatic resistance that P. aeruginosa presents, betalactamases, including extended spectrum (ESBL) and carbapenemases. But also aminoglycoside modifying enzymes, stand out, causing this microorganism to present multi-resistance phenotypes (MDR), extreme resistance (XDR) and pan-resistance (PDR) to the called antipseudomonal antibiotics, including the new cephalosporins with betalactamase inhibitors.

Prevalence of antibiotic resistance and virulent factors in nosocomial clinical isolates of Pseudomonas aeruginosa from Panamá

ABSTRACT Background: Pseudomonas aeruginosa is an important causative agent of nosocomial infections. As pathogen, P. aeruginosa is of increasing clinical importance due to its ability to develop high-level multidrug resistance (MDR). Methods: The aim of the present study was to better understand the intrinsic virulence of circulating strains of Pseudomonas aeruginosa, by surveying and characterizing the antibiotic resistance profiles and prevalence of virulence factors in 51 clinical isolates of P. aeruginosa obtained from children admitted to Hospital del Niño-Panamá during the period of October 2016 until March 2017. Antimicrobial susceptibilities were assessed by determining the minimum inhibitory concentration for 12 antibiotics against P. aeruginosa clinical isolates using the VITEK system (https://www.biomerieux.com). Additionally, all isolates were examined by Polymerase Chain Reaction (PCR) for the presence of components of the MexAB-OprM efflux pump system (mexABR) and pyoverdine receptor genes and betalactamases resistance genes (ESBL) using gene-specific primers. Results: A total of 51 pyoverdine producing clinical isolates were analyzed, all of which expressed resistance genes such as genes of the MexAB-OprM efflux pump system (mexABR) and pyoverdine receptor genes (fpvA). Out of 51 MDR isolates, 22 were ESBL producers. The most common ESBL gene was blaTEM expressed by 43% of the isolates. The isolates tested in this study showed increased resistance to antibiotics in the following categories: (i) penicillins (ampicillin (69%), piperacillin (22%); (ii) pyrimethamines (trimethoprim, 65%); (iii) nitrofurans (nitrofurantoin, 63%), and (iv) third-generation cephalosporin cefotaxime (53%). These results underscore a high prevalence of MDR amongst clinical isolates from Panama. Conclusions: The present study indicates that prevalence of BlaTEM-carrying strains is increasing with subsequent multidrug resistance in Panamá and as well reported worldwide. The virulent factors identified in this study provide valuable information regarding the prevalence of resistance genes and their potential impact on treatments that exploit the unique physiology of the pathogen. To prevent further spread of MDR, the proportions of resistant strains of Pseudomonas aeruginosa should be constantly evaluated on healthcare institutions of Panamá. More importantly, this information can be used to better understand the evolution and dissemination of strains hoping to prevent the development of resistance in Pseudomonas aeruginosa. Future studies quantifying the expression of these virulent genes will emphasize on the acquisition of multidrug resistance.

Clinical outcomes and molecular characterization of drug-resistant tuberculosis in pre- and extensively drug-resistant disease based on line probe assays

ABSTRACT Multidrug-resistant tuberculosis (MDR-TB) represents a significant impact in transmission, outcome, and health costs. The World Health Organization recommends implementation of rapid diagnostic methods for multidrug-resistance detection. This study was performed to evaluate the frequency of pre- and extensively drug resistant tuberculosis (pre-XDR-TB and XDR-TB) among MDR-TB patients, the pattern of resistance mutations for fluoroquinolones and the clinical outcome. Adult patients followed at a Brazilian regional reference center for TB, from January 2013 to June 2019 were included. Stored Mycobacterium tuberculosis (Mtb) cultures were recovered, the DNA was extracted, and the susceptibility test was performed using the line probe assay for second line antimycobacterial drugs, Genotype MTBDRsl version 2.0 (Hain Lifescience, CmbH, Germany). Among 33 MDR-TB included patients, we diagnosed XDR-TB or pre-XDR in five (15%) cases. Of these, mutations related to fluoroquinolones resistance were observed in four Mtb isolates, including one who had no phenotypic resistance profile. In two other patients with phenotypic resistance to ofloxacin, genotypic resistance was not found. Case fatality rate was 60% in pre/XDR-TB group, compared to 3.6% in the remaining of patients. This study observed few cases of pre-XDR and XDR-TB among a MDR-TB cohort. Phenotypic and genotypic assays presented good agreement. Clinical outcome was more favorable for patients with susceptibility to fluoroquinolones and injectable drugs.

A multidrug-resistant Stenotrophomonas maltophilia clinical isolate from Kamuzu Central Hospital, Malawi

Background Stenotrophomonas maltophilia is a significant opportunistic pathogen that is associated with high mortality in immunocompromised individuals. In this study, we describe a multidrug-resistant (MDR) S. maltophilia clinical isolate from Kamuzu Central Hospital (KCH), Lilongwe, Malawi. Methods: A ceftriaxone and meropenem nonsusceptible isolate (Sm-MW08), recovered in December 2017 at KCH, was referred to theNational Microbiology Reference Laboratory for identification. In April 2018, we identified the isolate using MALDI Biotyper mass spectrometry and determined its antimicrobial susceptibility profile using microdilution methods. Sm-MW08 was analysed by S1-PFGE, PCR, and Sanger sequencing, in order to ascertain the genotypes that were responsible for the isolate`s multidrug-resistance (MDR) phenotype. Results Sm-MW08 was identified as S. maltophilia and exhibited resistance to a range of antibiotics, including all ß-lactams, aminoglycosides (except arbekacin), chloramphenicol, minocycline, fosfomycin and fluoroquinolones, but remained susceptible to colistin and trimethoprim-sulfamethoxazole. The isolate did not harbour any plasmid but did carry chromosomally-encoded blaL1 metallo-ßlactamase and blaL2 ß-lactamase genes; this was consistent with the isolate's resistance profile. No other resistance determinants were detected, suggesting that the MDR phenotype exhibited by Sm-MW08 was innate. Conclusion : Herein, we have described an MDR S. maltophilia from KCH in Malawi, that was resistant to almost all locally available antibiotics. We therefore recommend the practice of effective infection prevention measures to curtail spread of this organism

First report of the aac(6)-Ib-cr gene in Providencia stuartii isolates in Brazil

Abstract INTRODUCTION: The aac(6')-Ib-cr and bla KPC genes are spreading among Enterobacteriaceae species, including Providencia stuartii, in some countries of world. METHODS: These genes were investigated in 28 P. stuartii isolates from a public hospital in Recife, Pernambuco, Brazil, by PCR and sequencing. RESULTS: The aac(6')-Ib-cr gene was detected in 16 resistant isolates, and the bla KPC gene was seen in 14. CONCLUSIONS: The presence of these genes in P. stuartii multi- and extensively drug-resistant isolates indicates that the resistance arsenal of this species is increasing, thus limiting the therapeutic options.

First report of a blaNDM-resistant gene in a Klebsiella aerogenes clinical isolate from Brazil

Abstract INTRODUCTION: Carbapenemase-resistant enterobacteria that produce the bla NDM gene are found worldwide. However, this is the first report of blaNDM in Klebsiella aerogenes in Brazil. METHODS: The identification of bacterial species was performed using anautomated system and confirmed by biochemical tests, 16S rRNA gene sequencing, and detection of resistance genes. RESULTS: The clinical isolate showed minimum inhibitory concentration resistance to meropenem and polymyxin B at 8mg/L and 4mg/L, respectively. Only the blaNDM gene was detected. CONCLUSIONS: The current report of the blaNDM gene in isolated MDR enterobacteria indicates that this gene can spread silently in a hospital setting.

Identification of a predominant genotype of Mycobacterium tuberculosis in Brazilian indigenous population

After nearly a century of vaccination and six decades of drug therapy, tuberculosis (TB) kills more people annually than any other infectious disease. Substantial challenges to disease eradication remain among vulnerable and underserved populations. The Guarani-Kaiowá people are an indigenous population in Paraguay and the Brazilian state of Mato Grosso do Sul. This community, marginalized in Brazilian society, experiences severe poverty. Like other South American indigenous populations, their TB prevalence is high, but the disease has remained largely unstudied in their communities. Herein, Mycobacterium tuberculosis isolates from local clinics were whole genome sequenced, and a population genetic framework was generated. Phylogenetics show M. tuberculosis isolates in the Guarani-Kaiowá people cluster away from selected reference strains, suggesting divergence. Most cluster in a single group, further characterized as M. tuberculosis sublineage 4.3.3. Closer analysis of SNPs showed numerous variants across the genome, including in drug resistance-associated genes, and with many unique changes fixed in each group. We report that local M. tuberculosis strains have acquired unique polymorphisms in the Guarani-Kaiowá people, and drug resistance characterization is urgently needed to inform public health to ensure proper care and avoid further evolution and spread of drug-resistant TB

Evaluation of Multidrug Resistant Loop-mediated Isothermal Amplification Assay for Detecting the Drug Resistance of / 生物医学与环境科学（英文）

Objective@#To evaluate multidrug resistant loop-mediated isothermal amplification (MDR-LAMP) assay for the early diagnosis of multidrug-resistant tuberculosis and to compare the mutation patterns associated with the @*Methods@#MDR-LAMP assay was evaluated using 100 @*Results@#The sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of MDR-LAMP were 85.5%, 93.6%, 96.7%, and 74.4% for the detection of resistance to isoniazid and rifampicin, respectively, and 80.5%, 92.3%, 98.6%, and 41.4% for the detection of @*Conclusion@#MDR-LAMP is a rapid and accessible assay for the laboratory identification of rifampicin and isoniazid resistance of

Infecciones del Tracto Urinario: métodos diagnósticos, tratamiento empírico y multirresistencia en una Unidad de Adultos Área de Emergencias / Urinary Tract Infections: diagnostic methods, empirical treatment and multidrug resistance in an Adult Emergency Area Unit

INTRODUCCIÓN. Las infecciones del tracto urinario por variedad de bacterias uropatógenas multiresistentes se deben al uso de tratamiento empírico o automedicación. OBJETIVO. Describir en las infecciones de tracto urinario los métodos diagnósticos, tratamiento empírico y la multirresistencia. MATERIALES Y MÉTODOS. Estudio observacional, descriptivo, retrospectivo. Población y muestra de 73 Historias Clínicas de pacientes atendidos en la Unidad de Adultos Área de Emergencias del Hospital de Especialidades Carlos Andrade Marín en el período enero a diciembre 2018. Se incluyeron pacientes mayores de 18 años, de ambos sexos, con diagnóstico clínico y por laboratorio de infección del tracto urinario superior e inferior. La información se obtuvo mediante la base de datos AS400, y se procesó en Epi-info y Excel. RESULTADOS. El 71,23% (52; 73) de mujeres tuvieron infección del tracto urinario. Escherichia coli fue frecuente en un 48,39% (15; 31), con mayor resistencia al Clotrimoxazol. El tratamiento empírico con Ciprofloxacino fue utilizado en 27,40% (20; 73). DISCUSIÓN: Se observó controversia en los tipos de estudios de imagen solicitados para el diagnóstico acorde a la clase de infección de tracto urinario así como el tratamiento empírico por factores propios de cada localidad que evitaron resistencia. CONCLUSIÓN. Escherichia coli se aisló de manera frecuente y registró mayor resistencia al Clotrimoxazol; el principal antibiótico prescrito como tratamiento empírico fue la Ciprofloxacina; el examen más solicitado fue la Urotomografía.

INTRODUCTION. Urinary tract infections due to a variety of multi-resistant uropathogenic bacteria are due to the use of empirical treatment or self-medication. OBJECTIVE. Describe diagnostic methods, empirical treatment and multidrug resistance in urinary tract infections. MATERIALS AND METHODS. Observational, descriptive, retrospective study. Population and sample of 73 Medical Records of patients treated in the Emergency Area Adult Unit of the Carlos Andrade Marín Specialty Hospital in the period january to december 2018. Patients older than 18 years of age, of both sexes, with clinical diagnosis and due to upper and lower urinary tract infection laboratory. The information was obtained through the AS400 database, and was processed in Epi-info and Excel. RESULTS. 71,23% (52; 73) of women had urinary tract infection. Escherichia coli was frequent in 48,39% (15; 31), with greater resistance to Clotrimoxazole. Empirical treatment with Ciprofloxacin was used in 27,40% (20; 73). DISCUSSION: Controversy was observed in the types of imaging studies requested for diagnosis according to the class of urinary tract infection as well as the empirical treatment due to factors specific to each locality that prevented resistance. CONCLUSION. Escherichia coli was frequently isolated and showed greater resistance to Clotrimoxazole; the main antibiotic prescribed as empirical treatment was Ciprofloxacin; the most requested examination was the Urotomography.

Antimicrobial resistance evaluation of bacteria isolated from infections in small animals in the Umuarama region, Paraná / Avaliação da resistência antimicrobiana de bactérias isoladas de infecções em pequenos animais na região de Umuarama, Paraná

Bacterial resistance is shown to be an inevitable side effect due to the excessive use of antibiotics, becoming a significant concern worldwide. Knowledge of regional bacterial resistance profiles enables the development of site-specific infection control practices, making conscious and moderate use of commercially available antibiotics. The aim of this study was the retrospective evaluation of the antimicrobial resistance profile of bacteria isolated from companion animal infections in the region of Umuarama/PR, from 2013 to 2017. This research was performed by analyzing the database belonging to the "Laboratório de Microbiologia Animal" at the "Universidade Estadual de Maringá" (UEM). Staphylococcus spp. represented 45.53% of the bacteria isolated from clinical infections in small animals in the period and place evaluated, followed by enterobacteria (34.04%), non-fermenting Gram-negative bacilli (NFGNB, 11.06%) and Streptococcus/Enterococcus (9.36%). A high number of antimicrobial resistance to antibiotics used in veterinary medicine was found. The lowest resistances associated with the best impact factor values were found for aminoglycosides, especially amikacin, chloramphenicol, and fluoroquinolones (norfloxacin and ciprofloxacin). Intermediate results were found for sulbactam-associated ampicillin, ceftriaxone, amoxicillin-clavulanic acid, and enrofloxacin. According to the number of resistant antimicrobial drugs, 64.26% (151/235) of the isolates were classified as multidrug-resistant, being 15.32% extensively resistant. Considering the resistance to antimicrobial classes, 68.94% (162/235) of the isolates were classified as multiresistant, being 19.15% extensively resistant. No bacterial strains were characterized as pan-resistant, but ten bacteria were resistant to all classes tested, with isolated susceptibility to certain drugs. Through the evaluation of resistance profiles found in the period and place studied and relevant literature, it is clear that there is a growing increase in the number of multiresistant bacteria among domestic animals which characterizes a serious risk to public health. The therapeutic arsenal is becoming increasingly diminished, and there is more difficulty in empirical drug selection, making antimicrobial susceptibility testing essential for more specific selection in antimicrobial therapy. Educational measures on the conscious use of antibiotics, infection control, and prevention of local specific zoonoses need to be instituted for the knowledge of health professionals and general access of the population.(AU)

A resistência bacteriana, mostra-se como um efeito colateral inevitável pelo excessivo uso de antibióticos, tornando-se alvo de grande preocupação mundial. O conhecimento dos perfis de resistência bacteriana regionais possibilita o desenvolvimento de práticas de controle de infecções específicas para cada localidade, fazendo uso consciente e moderado dos antibióticos disponíveis no mercado. O objetivo deste estudo foi a avaliação retrospectiva do perfil de resistência antimicrobiana de bactérias isoladas de infecções de animais de companhia na região de Umuarama/PR, no período de 2013 a 2017. Esta pesquisa foi realizada por meio da análise do banco de dados pertencente ao Laboratório de Microbiologia Animal da Universidade Estadual de Maringá (UEM). Os Staphylococcus spp. representaram 45,53% das bactérias isoladas de infecções clínicas em pequenos animais no período e local avaliado, seguido por enterobactérias (34,04%), bacilos Gram-negativos não fermentados (BGNNF, 11,06%) e Streptococcus/Enterococcus (9,36%). Um número elevado de resistência antimicrobiana frente aos antibióticos utilizados na medicina veterinária foi encontrado. As menores resistências associadas aos melhores valores do fator de impacto foram encontrados para aminoglicosídeos, em especial amicacina, cloranfenicol, fluoroquinolonas (norfloxacina e ciprofloxacina). Já resultados intermediários foram encontrados para ampicilina associada a sulbactam, ceftriaxona, amoxacilina com ácido clavulônico e enrofloxacina. Conforme o número de drogas antimicrobianas resistentes, foram classificados como multirresistentes 64,26% (151/235) dos isolados, sendo 15.32% extensivamente resistentes. Já considerando a resistência a classes de antimicrobianos, 68,94% (162/235) dos isolados foram classificados como multirresistentes, sendo 19.15% extensivamente resistentes. Nenhum isolado bacteriano foi caracterizado como pan-resistente, porém 10 bactérias foram resistentes a todas as classes testadas, com susceptibilidade isolada a determinadas drogas. Por meio da avaliação dos perfis de resistência encontrados no período e local estudados e de literatura pertinente, percebe-se que há um aumento crescente no número de bactérias multirresistentes entre os animais domésticos o que caracteriza um grave risco à saúde pública. O arsenal terapêutico está se tornando cada vez mais diminuto e há mais dificuldade na seleção empírica de drogas, tornando essencial a realização de testes de susceptibilidade antimicrobiana para uma seleção mais específica na terapêutica antimicrobiana. Medidas educativas sobre o uso consciente dos antibióticos, controle de infecções e prevenção de zoonoses específicas para as localidades precisam ser instituídas para conhecimento dos profissionais do setor da saúde e acesso geral da população.(AU)