This report presents the first evidence of autochthonous circulation of the NDM-1 resistance gene harbored by an IncA/C plasmid isolated from a K. pneumoniae ST147 in Ecuador. Efforts should be implemented to monitor and characterize the spatial and temporal distribution of NDM in Ecuador and other countries of South America.
Contexto: Escherichia coli uropatógena (ECUP) se presenta como uno de los principales agentes etiológicos en infecciones del tracto urinario (ITUs) no complicadas (70-95%). El objetivo del tratamiento de ITUs no complicadas es obtener curación clínica y microbiológica. Para ello, es de particular importancia el conocimiento de las tasas de resistencia antibiótica local.Objetivo: identificar los perfiles de resistencia a antibióticos de primera línea para ITUs no complicadas en poblaciones nativas amerindias Kichwas ecuatorianas, en donde el tratamiento empírico se basa en trimetoprim/sulfametoxazol, ampicilina, y ciprofloxacina mayoritariamente.Métodos: se analizaron 335 muestras de orina procedentes de las poblaciones de Zumbahua, Colta y Guamote, en un periodo de 4 meses (febrero-mayo 2016). Las muestras fueron incubadas por 24 y 48 horas en agar Eosin Methylene Blue (EMB), para luego ser identificadas en género y especie por pruebasbioquímicas. Para determinar la susceptibilidad antibiótica, se realizó la técnica de difusión en disco de Kirby-Bauer. Para la Concentración Inhibitoria Mínima (CIM), se utilizó la técnica de microdilución en caldo (Vitek 2). El método de doble disco fue la técnica utilizada para la detección de betalactamasas deespectro extendido (BLEE).Resultados: noventa (26,9%) muestras mostraron un recuento significativo de ≥105 (ufc)/ml, compatibles con ITUs. El microorganismo identificado con mayor frecuencia fue E. coli (n=75; 83,3%). La resistencia antibiótica encontrada para los aislados de E. coli fue de 56,7% a trimetoprim/sulfametoxazol, 52,5% aampicilina, 43.3% a ácido nalidíxico, 32.5% a ciprofloxacina, 28.3% a norfloxacina, 25% a levofloxacina, 15.85% a cefazolina, 17.5% a cefoxitina, 15% a cefuroxima, 15% a ceftazidima, cefotaxima, y ceftriaxona, 15% a cefepima, 7,5% a nitrofurantoina y 1,7% a fosfomicina. Se identificaron 7 aislados productores de betalactamasas de espectro extendido (BLEE).Conclusión: con los resultados obtenidos se recomienda no utilizar ampicilina, trimetoprim/sulfametoxazol, ni quinolonas en la zona estudiada como terapia empírica. Se sugiere instaurar tratamiento empírico con fosfomicina o nitrofurantoina para ITUs no complicadas
23The aim of this study was to investigate the presence of Escherichia coli carrying mcr-1 gene 24 in domestic animals close to a child who suffered a peritoneal infection by a mcr-1 positive 25 E. coli. Rectal or cloacal swabs and fecal samples from domestic animals were plated on 26 selective media to isolate colistin-resistant E. coli and isolates were submitted to detection of 27 mcr-1 gene, pulsed field gel electrophoresis (PFGE), multi-locus sequence typing (MLST), 28 replicon typing and S1-PFGE. Four mcr-1 positive E. coli isolates (from chicken, turkey and 29 dog) were recovered. No shared PFGE pattern or MLST sequence type were observed among 30 isolates. A 60Kb IncI1 mcr-1-carrying plasmid was detected in all isolates. Our results 31 suggest that mcr-1 gene was horizontally disseminated amongst different lineages of E. coli 32 from domestic animals in the child's household. 33 Importance 34Horizontally transferable colistin resistance (mcr-1 gene) is thought to have originated in 35 domestic animals and transferred to humans through meat and dairy products. In the present 36 report we show evidence that the mcr-1 gene could be transferred to different E. coli strains 37 colonizing different hosts (humans and pets) in the same household. 38 42 different bacterial genera colonizing different animal species (2, 3).43 3The first report of a colistin resistance (CR) gene carried by plasmids (mcr-1) came from China in 44 2015 (4). This gene codes for a phosphoethanolamine transferase (MCR-1), which modifies the 45 lipid A moiety in the outer membrane of Gram negative bacteria and confers resistance to 46 polymyxins (4, 5). Among Enterobacteriacea different mcr gen groups (1-5) could be transferred by 47 PCR was performed to detect mcr gene (4); amplicons were sequence and aligned using mcr-1 130 (NG_055582.1), mcr-2 (NG_051171.1), mcr-3 ( NG_056184.1), mcr-4( MG822665.1), mcr-5 131 (MG384740.1) accession numbers with Geneius software. Pulsed field gel electrophoresis (PFGE) 132 (25) and multilocus sequence typing (MLST) was performed on seven housekeeping genes to 133 define clonal relatedness (26). Replicon typing was performed using a commercial kit (PBRT KIT, 134 DIATHE, Fano, Italy) (20, 27, 28). -lactamase genes (blaCTX-M-1, blaTEM, blaSHV) were detected 135 using primers previously described (29). Briefly, 12,5mL of GoTaq ® Green Master Mix 136 (Promega, Madison, USA) were mixed with 1L of upstream primer, 10M, 1L of downstream 137 primer, 10 L of DNA template and 9,5 L of Nuclease-free water to complete a 25L of 138 reaction volume. The reaction mix were amplified using 2-minute of initial denaturation at 94°C 139 followed by 40 cycles of DNA denaturation at 94°C (40 sec), annealing at 60°C (40sec) and 140 extension at 72°C (1min). The final elongation step at 72°C for 5 min. Amplicons were detected by 141 electrophoresis in a 2% agarose gel. For complete amplification and sequencing of the detected 142resistance genes, primers and conditions previously described were used (30). 143 S1-PFG...
Objective. To compare the epidemiology of antimicrobial resistance in bacteria isolated from inpatient and outpatient samples in Ecuador. Methods. A secondary analysis was done of data on bacteria isolated from inpatient and outpatient samples. Data were taken from the 2018 national antimicrobial resistance surveillance database of the National Reference Center for Antimicrobial Resistance. The variables included were: age, sex, inpatient versus outpatient setting, type of specimen, bacterial species identified, pattern of resistance to antibiotics, and geographic area. Results. Data from 57 305 bacterial isolates were included in the study: 48.8% were from hospitalized patients, 55.7% were from women, and 60.1% were from patients older than 45 years. Urine (42.9%) and blood (12.4%) were the most common clinical samples. Overall, 77.1% of bacterial isolates were gram-negative (83% and 71% in outpatients and inpatients, respectively). The most common gram-positive and gram-negative species were Staphylococcus aureus and Escherichia coli, respectively. Antimicrobial resistance levels were high (up to 80% for some antimicrobial drugs), and were higher in hospitalized patients compared with outpatients. A variety of carbapenemases were found to confer resistance to carbapenems (antibiotics of last resort) in gram-negative bacteria. Conclusions. The study findings provide an important baseline on antimicrobial resistance in Ecuador. This will allow the strengthening of guidelines of the surveillance system, the creation of public policies for standardization of laboratory methodologies, the proper handling of information, and the development of empirical therapy guidelines based on local epidemiology.
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