Molecular epidemiology of carbapenem-resistant gram-negative bacilli in Ecuador.

INTRODUCTION: Carbapenem-resistant gram-negative bacilli are a worldwide concern because of high morbidity and mortality rates. Additionally, the increasing prevalence of these bacteria is dangerous. To investigate the extent of antimicrobial resistance and prioritize the utility of novel drugs, we evaluated the molecular characteristics and antimicrobial susceptibility profiles of carbapenem-resistant Enterobacterales, Pseudomonas aeruginosa and Acinetobacter baumannii in Ecuador in 2022. METHODS: Ninety-five clinical isolates of carbapenem non-susceptible gram-negative bacilli were collected from six hospitals in Ecuador. Carbapenem resistance was confirmed with meropenem disk diffusion assays following Clinical Laboratory Standard Institute guidelines. Carbapenemase production was tested using a modified carbapenemase inactivation method. Antimicrobial susceptibility was tested with a disk diffusion assay, the Vitek 2 System, and gradient diffusion strips. Broth microdilution assays were used to assess colistin susceptibility. All the isolates were screened for the blaKPC, blaNDM, blaOXA-48, blaVIM and blaIMP genes. In addition, A. baumannii isolates were screened for the blaOXA-23, blaOXA-58 and blaOXA-24/40 genes. RESULTS: Carbapenemase production was observed in 96.84% of the isolates. The blaKPC, blaNDM and blaOXA-48 genes were detected in Enterobacterales, with blaKPC being predominant. The blaVIM gene was detected in P. aeruginosa, and blaOXA-24/40 predominated in A. baumannii. Most of the isolates showed co-resistance to aminoglycosides, fluoroquinolones, and trimethoprim/sulfamethoxazole. Both ceftazidime/avibactam and meropenem/vaborbactam were active against carbapenem-resistant gram-negative bacilli that produce serin-carbapenemases. CONCLUSION: The epidemiology of carbapenem resistance in Ecuador is dominated by carbapenemase-producing K. pneumoniae harbouring blaKPC. Extensively drug resistant (XDR) P. aeruginosa and A. baumannii were identified, and their identification revealed the urgent need to implement strategies to reduce the dissemination of these strains.

Emergence of Raoultella ornithinolytica in human infections from different hospitals in Ecuador with OXA-48-producing resistance.

Purpose: The purpose of this study was to highlight the clinical and molecular features of 13 Raoultella ornithinolytica strains isolated from clinical environments in Ecuador, and to perform comparative genomics with previously published genomes of Raoultella spp. As Raoultella is primarily found in environmental, clinical settings, we focused our work on identifying mechanisms of resistance that can provide this bacterium an advantage to establish and persist in hospital environments. Methods: We analyzed 13 strains of Raoultella ornithinolytica isolated from patients with healthcare associated infections (HAI) in three hospitals in Quito and one in Santo Domingo de Los Tsáchilas, Ecuador, between November 2017 and April 2018. These isolates were subjected to phenotypic antimicrobial susceptibility testing, end-point polymerase chain reaction (PCR) to detect the presence of carbapenemases and whole-genome sequencing. Results: Polymerase chain reaction revealed that seven isolates were positive isolates for blaOXA-48 and one for blaKPC-2 gene. Of the seven strains that presented the blaOXA-48 gene, six harbored it on an IncFII plasmid, one was inserted into the bacterial chromosome. The blaKPC gene was detected in an IncM2/IncR plasmid. From the bioinformatics analysis, nine genomes had the gene blaOXA-48, originating from Ecuador. Moreover, all R. ornithinolytica strains contained the ORN-1 gene, which confers resistance for ß-lactams, such as penicillins and cephalosporins. Comparative genome analysis of the strains showed that the pangenome of R. ornithinolytica is considered an open pangenome, with 27.77% of core genes, which could be explained by the fact that the antibiotic resistance genes in the ancestral reconstruction are relatively new, suggesting that this genome is constantly incorporating new genes. Conclusion: These results reveal the genome plasticity of R. ornithinolytica, particularly in acquiring antibiotic-resistance genes. The genomic surveillance and infectious control of these uncommon species are important since they may contribute to the burden of antimicrobial resistance and human health.

Detection and genotyping of Listeria monocytogenes in artisanal soft cheeses from Ecuador

Abstract Listeria monocytogenes is one of the most important bacteria associated with food-borne diseases, soft cheese being an important L. monocytogenes vehicle. In Ecuador, softcheese is consumed in 84.3% of urban households. We determined the prevalence of L. mono-cytogenes and serogroups in 260 fresh artisanal soft cheese samples collected in 18 of 24Ecuadorian provinces. Listeria spp. detection was carried out by culture-dependent and inde-pendent methods; 14.23% of samples were positive for L. monocytogenes. Serogroup IVb wasfound in 83.78% of the food isolates. Serogroups IIb and IIa were present in 8.11% of our isolates.To our knowledge, this is the first report of L. monocytogenes serogroups associated with foodin Ecuador; we also found serogroup similarities between cheese isolates and clinical isolates.

Resumen Listeria monocytogenes es una de las bacterias más importantes asociadas conenfermedades transmitidas por alimentos, y el queso fresco es un importante vehículo de trans-misión de L. monocytogenes. En Ecuador, el consumo de quesos frescos se produce en el 84,3%de los hogares urbanos. Determinamos la prevalencia de L. monocytogenes y sus serogrupos en260 muestras de queso fresco artesanal recolectadas en 18 de las 24 provincias ecuatorianas.La detección de Listeria spp. se llevó a cabo mediante métodos independientes y dependientesde cultivo; el 14,23% de las muestras fueron positivas para L. monocytogenes.

Detection and genotyping of Listeria monocytogenes in artisanal soft cheeses from Ecuador.

Listeria monocytogenes is one of the most important bacteria associated with foodborne diseases, soft cheese being an important L. monocytogenes vehicle. In Ecuador, soft cheese is consumed in 84.3% of urban households. We determined the prevalence of L. monocytogenes and serogroups in 260 fresh artisanal soft cheese samples collected in 18 of 24 Ecuadorian provinces. Listeria spp. detection was carried out by culture-dependent and independent methods; 14.23% of samples were positive for L. monocytogenes. Serogroup IVb was found in 83.78% of the food isolates. Serogroups IIb and IIa were present in 8.11% of our isolates. To our knowledge, this is the first report of L. monocytogenes serogroups associated with food in Ecuador; we also found serogroup similarities between cheese isolates and clinical isolates.

Characterization of the genetic structure of mcr-1 gene among Escherichia coli isolates recovered from surface waters and sediments from Ecuador.

Although anthropogenic activities contribute to the selection and spread of antibiotic resistance in aquatic environments, limited information is available from countries with absent or incomplete sewage treatment systems and the impact of their discharges onto water bodies. This study therefore aimed to characterize the genetic structure of colistin resistance (mcr) genes among Escherichia coli isolates recovered from surface waters and sediments in Ecuador. Out of 459 isolates, four Escherichia coli showed multidrug-resistant phenotypes, which harbored the mcr-1 gene and ß-lactamases, such as blaTEM, blaCTX-M-15, blaCTX-M-55, or blaCTX-M-65 genes. Three E. coli isolates (U20, U30 and U144) shared a similar genetic environment surrounding the mcr-1 gene, which was located on plasmids. Only one E. coli isolate (U175) showed that the mcr-1 gene was chromosomally located. Moreover, the core genome multilocus sequence typing (cgMLST) analysis revealed that these isolates belong to different lineages. This study represents the first detection of the mcr-1 gene in multidrug-resistant E. coli isolates from environmental samples in Ecuador.

Bordetella pertussis, a reemerging pathogen in pediatric respiratory infections. A study in Quito, Ecuador.

Bordetella pertussis is the causative agent of pertussis, which mainly affects unvaccinated children, while Bordetella parapertussis causes a disease presenting clinical characteristics that are indistinguishable from whooping cough. Despite high vaccination coverage, pertussis remains a public health concern worldwide, with approximately 140000 cases reported annually. Here we determined the prevalence of B. pertussis and B. parapertussis infection among infants under one year of age by polymerase chain reaction (PCR); our aim being to identify whether the data obtained relates to the relevant sociodemographic and clinical data. The study included 86 samples of nasopharyngeal swabs from infants aged between 0-12 months, who were reported as probable cases of whooping cough by the health centers around the Ecuadorian highlands, from August 2016 to July 2017. The nasopharyngeal swabs were cultured and microbiological and molecular analyses were performed. B. pertussis was identified by PCR in 41% of the samples (30/86), more than half of which corresponded to infants aged between 0-3 months. Moreover, a statistically significant correlation (p<0.05) between the identification of bacteria in culture and the catarrhal stage of the disease was observed. The results obtained from the study highlighted the need for an active national surveillance of pertussis, in particular for laboratory testing, to provide a highly sensitive and more specific diagnosis of Bordetella infection.

Broiler Farms and Carcasses Are an Important Reservoir of Multi-Drug Resistant Escherichia coli in Ecuador.

Antimicrobial resistance (AMR) is a major health threat for public and animal health in the twenty-first century. In Ecuador, antibiotics have been used by the poultry industry for decades resulting in the presence of multi-drug resistant (MDR) bacteria in the poultry meat production chain, with the consequent risk for public health. This study evaluated the prevalence of ESBL/AmpC and mcr genes in third-generation cephalosporin-resistant Escherichia coli (3GC-R E. coli) isolated from broiler farms (animal component), broiler carcasses (food component), and human enteritis (human component) in Quito-Ecuador. Samples were collected weekly from November 2017 to November 2018. For the animal, food, and human components, 133, 335, and 302 samples were analyzed, respectively. Profiles of antimicrobial resistance were analyzed by an automated microdilution system. Resistance genes were studied by PCR and Sanger sequencing. From all samples, 122 (91.7%), 258 (77%), and 146 (48.3%) samples were positive for 3GC-R E. coli in the animal, food, and human components, respectively. Most of the isolates (472/526, 89.7%) presented MDR phenotypes. The ESBL bla CTX-M-55, bla CTX-M-3, bla CTX-M-15, bla CTX-M-65, bla CTX-M-27, and bla CTX-M-14 were the most prevalent ESBL genes while bla CMY-2 was the only AmpC detected gene. The mcr-1 gene was found in 20 (16.4%), 26 (10.1%), and 3 (2.1%) of isolates from animal, food, and human components, respectively. The implication of poultry products in the prevalence of ESBL/AmpC and mcr genes in 3GC-R must be considered in the surveillance of antimicrobial resistance.

Comparación de dos métodos comerciales para la determinación de la concentración mínima inhibitoria de meropenem en Klebsiella pneumoniae productora de carbapenemasa tipo KPC / Comparison of two commercial methods for determination of the minimum inhibitory meropenem concentration in KPC carbapenemase-producing Klebsiella pneumoniae

Introducción: El tratamiento de las infecciones por Klebsiella pneumoniae productora de carbapenemasa tipo KPC es complicado debido a las escasas opciones terapéuticas existentes, lo cual obliga a optimizar los esquemas terapéuticos disponibles. Objetivo: Determinar la concordancia de la tarjeta AST-N272 del Sistema Vitek 2 Compact y las tiras M.I.C.ETM Evaluator con la dilución en agar para la determinación de la concentración mínima inhibitoria del meropenem en Klebsiella pneumoniae productora de carbapenemasa tipo KPC. Métodos: Se estudiaron 53 aislados de K. pneumoniae bla KPC positivas no clonales, provenientes de hisopados rectales recolectados en diferentes unidades hospitalarias de Guayaquil, Ecuador, entre enero a junio de 2016. Se determinó la concentración mínima inhibitoria de meropenem por dilución en agar (método de referencia), así como por el sistema Vitek 2 Compact (AST-N272) y las tiras M.I.C.ETM. Se determinó la CMI 50, CMI 90 y la concordancia esencial. Resultados: El rango de la CMI de meropenem de los aislados estudiados fue de 1 a ≥ 32 µg/mL, con una CMI50= 4 µg/mL y una CMI90= ≥ 32 µg/mL. El 86,79 por ciento (n= 46) de los aislados tuvo una CMI≤ 8 µg/mL. Se observó un 94,33 por ciento de concordancia esencial con las tiras M.I.C.ETM, mientras que la tarjeta AST-N272 mostró una concordancia esencial inferior al 50 por ciento. Conclusiones: Los resultados sugieren posibles implicaciones en el tratataminto del paciente, pues reduce opciones terapéuticas en contextos de difícil manejo. Además, resaltan la necesidad de la confirmación de la resistencia a carbapenémicos mediante el método de Kirby Bawer en aquellos laboratorios que tienen métodos automatizados para estudios de susceptibilidad(AU)

Introduction: The treatment for KPC carbapenemase-producing Klebsiella pneumoniae infections is complicated, due to the scant therapeutic options available, which forces us to optimize the therapies at hand. Objective: Determine the agreement between the AST-N272 card of the Vitek 2 Compact system and the M.I.C.E.TM Evaluator strips, and the agar dilution method for determination of the minimum inhibitory meropenem concentration in KPC carbapenemase-producing Klebsiella pneumoniae. Methods: A study was conducted of 53 positive non-clonal K. pneumoniae bla KPC isolates from rectal swabs collected at several hospitals in Guayaquil, Ecuador, from January to June 2016. Minimum inhibitory meropenem concentration was determined by agar dilution (reference method), the Vitek 2 Compact system (AST-N272) and M.I.C.E.TM strips. Determination was made of MIC 50, MIC 90 and essential agreement. Results: The meropenem MIC range for the isolates studied was 1 to ≥ 32 µg/ml, with MIC50= 4 µg/ml and MIC90= ≥ 32 µg/ml. In 86.79 percent (n= 46) of the isolates MIC was ≤ 8 µg/ml. Essential agreement was 94.33 percent with the M.I.C.E.TM strips and under 50 percent with the AST-N272 card. Conclusions: The results obtained suggest potential implications for the treatment of patients, since therapeutic options are reduced in difficult management contexts. They also highlight the need for confirmation of carbapenem resistance by the Kirby-Bauer procedure in laboratories equipped with automated methods for susceptibility studies(AU)

Fosfomycin, Applying Known Methods and Remedies to A New Era.

The exponential increase in the numbers of isolates of Carbapenem-Resistant Enterobacteriaceae (CRE) creates the need for using novel therapeutic approaches to save the lives of patients. Fosfomycin has long been considered a rational option for the treatment of CRE to be used as part of a combined therapy scheme. However, the assessment of fosfomycin susceptibility in the laboratory presents a great challenge due to the discrepancies found between different methodologies. Thus, our goal was to evaluate fosfomycin susceptibility in a group of 150 Enterobacteriaceae bacterial isolates using agar dilution as the gold standard technique to compare the results with those obtained by disk diffusion. We found a fosfomycin susceptibility of 79.3% in general terms. By comparing both methodologies, we reported a categorical agreement of 96% without Very Major Errors (VMEs) or Major Errors (MEs) and 4% of minor Errors (mEs). Our results suggest that fosfomycin could provide a rational alternative treatment for those patients that are infected by a Multidrug-Resistant (MDR) microorganism that is currently untreatable and that the disk diffusion and classical agar dilution techniques are adequate to assess the resistance profile of CRE to fosfomycin.

OXA-48 Carbapenemase in Klebsiella pneumoniae Sequence Type 307 in Ecuador.

Antibiotic resistance is on the rise, leading to an increase in morbidity and mortality due to infectious diseases. Klebsiella pneumoniae is a Gram-negative bacterium that causes bronchopneumonia, abscesses, urinary tract infection, osteomyelitis, and a wide variety of infections. The ubiquity of this microorganism confounds with the great increase in antibiotic resistance and have bred great concern worldwide. K. pneumoniae sequence type (ST) 307 is a widespread emerging clone associated with hospital-acquired infections, although sporadic community infections have also been reported. The aim of our study is to describe the first case of Klebsiella pneumoniae (ST) 307 harboring the blaOXA-48-like gene in Ecuador. We characterized a new plasmid that carry OXA-48 and could be the source of future outbreaks. The strain was recovered from a patient with cancer previously admitted in a Ukrainian hospital, suggesting that this mechanism of resistance could be imported. These findings highlight the importance of programs based on active molecular surveillance for the intercontinental spread of multidrug-resistant microorganisms with emergent carbapenemases.

Efficacy of pulsed-xenon ultraviolet light for disinfection of high-touch surfaces in an Ecuadorian hospital.

BACKGROUND: Hospital environment in patient care has been linked on healthcare-associated infections (HAI). No touch disinfection technologies that utilize pulsed xenon ultraviolet light has been recognized to prevent infection in contaminated environments. The purpose of this study was: 1) to evaluate the effectiveness of pulsed-xenon ultraviolet light (PX-UV) disinfection for the reduction of bacteria on environmental surfaces of Hospital General Enrique Garcés, and 2) to evaluate the in-vitro efficacy against multi-drug resistance microorganisms. METHODS: This was a quality-improvement study looking at cleaning and disinfection of patient areas. During the study, a total of 146 surfaces from 17 rooms were sampled in a secondary 329-bed public medical center. Microbiological samples of high-touch surfaces were taken after terminal manual cleaning and after pulsed xenon ultraviolet disinfection. Cleaning staff were blinded to the study purpose and told clean following their usual protocols. For positive cultures PCR identification for carbapenemase-resistance genes (blaKPC, blaIMP, blaVIM, and blaNDM) were analyzed and confirmed by sequencing. The total number of colony forming units (CFU) were obtained and statistical analyses were conducted using Wilcoxon Rank Sum tests to evaluate the difference in CFU between terminal manual cleaning and after pulsed xenon ultraviolet disinfection. RESULTS: After manual disinfection of 124 surfaces showed a total of 3569 CFU which dropped to 889 CFU in 80 surfaces after pulsed xenon disinfection (p < 0.001). Overall, the surface and environmental contamination was reduced by 75% after PX-UV compared to manual cleaning and disinfection. There were statistically significant decreases in CFU counts of high touch surfaces in OR 87% (p < 0.001) and patient rooms 76% (p < 0.001). Four rooms presented serine carbapenemases blaKPC, and metallo beta-lactamases blaNDM, blaVIM, blaIMP. confirmed by PCR and sequencing. The in-vitro testing with endemic strains found that after five minutes of pulsed xenon ultraviolet exposure an 8-log reduction was achieved in all cases. CONCLUSION: This study is one of the first of its kind in an Ecuador Hospital. We found that pulsed-xenon ultraviolet disinfection technology is an efficacious complement to the established manual cleaning protocols and guidelines in the significant reduction of MDRO.

First case of New Delhi metallo-ß-lactamase in Klebsiella pneumoniae from Ecuador: An update for South America.

OBJECTIVES: To describe a clinical case of Klebsiella pneumoniae harboring a New Delhi metallo-ß-lactamase (NDM) plasmid in Ecuador and to present a map of reports of NDM isolates in South America. METHODS: The modified Hodge test, carbapenem inactivation method, imipenem-EDTA disk method (synergy), and Rapidec Carba NP test were used to identify antibiotic resistance mechanisms. The presence of resistance genes was explored with a conjugation assay, and molecular confirmation of NDM was performed by PCR and DNA sequencing. Plasmid characterization was conducted by PCR-based replicon typing. A literature review was performed in Google Scholar and PubMed to identify reports from South America. RESULTS: An HIV-infected patient, who had never traveled abroad, developed a bloodstream infection caused by K. pneumoniae ST147 harboring the NDM-1 resistance gene in a plasmid from the IncA/C group. Local circulation of NDM has also been described in other South American countries, in particular in Colombia and Brazil, although published scientific records were not found for other countries. CONCLUSIONS: 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.

Resistencia bacteriana de Escherichia coli uropatogénica en población nativa amerindia Kichwa de Ecuador / Bacterial resistance of uropathogenic Escherichia coli in native Kichwa Amerindian population of Ecuador

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 pruebas bioquímicas. 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 de espectro 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% a ampicilina, 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. (AU)

Brote por Serratia marcescens en una Unidad de Cuidados Intensivos Neonatales: Guayaquil-Ecuador / Serratia marcescens outbreak in Neonatal Intensive Care Unit: Guayaquil, Ecuador

We report a Serratia marcescens outbreak occurred in the NICU of a pediatric hospital in Guayaquil, Ecuador. Nine cases of infection were detected, from which septicemia was developed in 55.5%. The index case was a newborn derived from another institution with septic arthritis caused by the outbreak strain. The infection rate was 17.6% and mortality rate was 33.3%. All isolates were resistant to aminoglycosides and susceptible to third generation cephalosporins and carbapenems. Clonality analysis by pulsed-field gel electrophoresis (PFGE) revealed the presence of two closely related clones confirming the horizontal spread. Measures were taken by the committee such as: strengthening the hand hygiene, patient hygiene and cohort studies of gastrointestinal colonization, which allowed the control of the outbreak.

Comunicamos un brote nosocomial por Serratia marcescens en una Unidad de Cuidados Intensivos en un hospital pediátrico de Guayaquil, Ecuador. Se detectaron nueve casos de infección, manifestándose en 55,5% de los casos como sepsis. El caso índice correspondió a un neonato derivado de otra institución con artritis séptica. La tasa de ataque fue 17,6% (n: 51) y la mortalidad 33,3%. Todos los aislados presentaron resistencia a las cefalosporinas y aminoglucósidos y sensibilidad a carbapenémicos. El análisis de clonalidad reveló la presencia de dos clones estrechamente relacionados, confirmando la diseminación horizontal. Las medidas de control de brote fueron: reforzamiento de higiene de manos, cohorte de los pacientes y búsqueda de colonización gastrointestinal.

[Serratia marcescens outbreak in Neonatal Intensive Care Unit: Guayaquil, Ecuador]. / Brote por Serratia marcescens en una Unidad de Cuidados Intensivos Neonatales: Guayaquil-Ecuador.

We report a Serratia marcescens outbreak occurred in the NICU of a pediatric hospital in Guayaquil, Ecuador. Nine cases of infection were detected, from which septicemia was developed in 55.5%. The index case was a newborn derived from another institution with septic arthritis caused by the outbreak strain. The infection rate was 17.6% and mortality rate was 33.3%. All isolates were resistant to aminoglycosides and susceptible to third generation cephalosporins and carbapenems. Clonality analysis by pulsed-field gel electrophoresis (PFGE) revealed the presence of two closely related clones confirming the horizontal spread. Measures were taken by the committee such as: strengthening the hand hygiene, patient hygiene and cohort studies of gastrointestinal colonization, which allowed the control of the outbreak.