Molecular characterization of carbapenemase-producing Enterobacterales in a tertiary hospital in Lima, Peru.

Carbapenemase-producing Enterobacterales (CPE) are a growing threat to global health and the economy. Understanding the interactions between resistance and virulence mechanisms of CPE is crucial for managing difficult-to-treat infections and informing outbreak prevention and control programs. Here, we report the characterization of 21 consecutive, unique clinical isolates of CPE collected in 2018 at a tertiary hospital in Lima, Peru. Isolates were characterized by phenotypic antimicrobial susceptibility testing and whole-genome sequencing to identify resistance determinants and virulence factors. Seven Klebsiella pneumoniae isolates were classified as extensively drug-resistant. The remaining Klebsiella, Enterobacter hormaechei, and Escherichia coli isolates were multidrug-resistant. Eighteen strains carried the metallo-ß-lactamase NDM-1, two the serine-carbapenemase KPC-2, and one isolate had both carbapenemases. The blaNDM-1 gene was located in the truncated ΔISAba125 element, and the blaKPC-2 gene was in the Tn4401a transposon. ST147 was the most frequent sequence type among K. pneumoniae isolates. Our findings highlight the urgent need to address the emergence of CPE and strengthen control measures and antibiotic stewardship programs in low- and middle-income settings.IMPORTANCEGenomic surveillance of antimicrobial resistance contributes to monitoring the spread of resistance and informs treatment and prevention strategies. We characterized 21 carbapenemase-producing Enterobacterales collected at a Peruvian tertiary hospital in 2018, which exhibited very high levels of resistance and carried numerous resistance genes. We detected the coexistence of carbapenemase-encoding genes (blaNDM-1 and blaKPC-2) in a Klebsiella pneumoniae isolate that also had the PmrB(R256G) mutation associated with colistin resistance. The blaKPC-2 genes were located in Tn4401a transposons, while the blaNDM-1 genes were in the genetic structure Tn125 (ΔISAba125). The presence of high-risk clones among Klebsiella pneumoniae (ST11 and ST147) and Escherichia coli (ST410) isolates is also reported. The study reveals the emergence of highly resistant bacteria in a Peruvian hospital, which could compromise the effectiveness of current treatments and control.

First Report of OXA-181-Producing Enterobacterales Isolates in Latin America.

We characterized five carbapenemase-producing Enterobacterales (CPE) isolates from two health care institutions in Lima, Peru. The isolates were identified as Klebsiella pneumoniae (n = 3), Citrobacter portucalensis (n = 1), and Escherichia coli (n = 1). All were identified as blaOXA-48-like gene carriers using conventional PCR. Whole-genome sequencing found the presence of the blaOXA-181 gene as the only carbapenemase gene in all isolates. Genes associated with resistance to aminoglycosides, quinolones, amphenicols, fosfomycins, macrolides, tetracyclines, sulfonamides, and trimethoprim were also found. The plasmid incompatibility group IncX3 was identified in all genomes in a truncated Tn6361 transposon flanked by ΔIS26 insertion sequences. The qnrS1 gene was also found downstream of blaOXA-181, conferring fluoroquinolone resistance to all isolates. CPE isolates harboring blaOXA-like genes are an increasing public health problem in health care settings worldwide. The IncX3 plasmid is involved in the worldwide dissemination of blaOXA-181, and its presence in these CPE isolates suggests the wide dissemination of blaOXA-181 in Peru. IMPORTANCE Reports of carbapenemase-producing Enterobacterales (CPE) isolates are increasing worldwide. Accurate detection of the ß-lactamase OXA-181 (a variant of OXA-48) is important to initiate therapy and preventive measures in the clinic. OXA-181 has been described in CPE isolates in many countries, often associated with nosocomial outbreaks. However, the circulation of this carbapenemase has yet to be reported in Peru. Here, we report the detection of five multidrug-resistant CPE clinical isolates harboring blaOXA-181 in the IncX3-type plasmid, a potential driver of dissemination in Peru.

Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp / In vitro antimicrobial activity of Bixa orellana L. leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp.

Objetivos . Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. Materiales y métodos . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. Resultados . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). Conclusiones . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Objective. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. Materials and methods. Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. Results. All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. Conclusions. In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

In vitro antimicrobial activity of Bixa orellana L . Leaves extract against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. / Actividad antimicrobiana in vitro del extracto de hojas de Bixa orellana L. contra bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp.

OBJECTIVE.: Motivation for the study: bacterial vaginosis is a bacterial infection that frequently affects women of reproductive age. The treatment is based on synthetic antimicrobials. Bixa orellana L. possesses antimicrobial properties and could represent a potential non-synthetic therapeutic alternative. Main findings: in vitro results suggest that, methanolic extract of Bixa orellana L. leaves possesses potential antimicrobial properties against bacteria associated to bacterial vaginosis. Implications: to identify new sources with therapeutic potential, and to promote research, discovery, and characterization of non-synthetic antimicrobials. To describe the in vitro antimicrobial activity of the methanolic extract of Bixa orellana L. leaves against anaerobic bacteria associated to bacterial vaginosis and Lactobacillus spp. MATERIALS AND METHODS.: Eight ATCC reference strains; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula, and Lactobacillus crispatus, and twenty-two clinical isolates; eleven Gardnerella vaginalis and eleven Lactobacillus strains, were included in the study. The antimicrobial susceptibility was determined by the agar diffusion method. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined by using agar dilution and a modified dilution plating method, respectively. RESULTS.: All ATCC reference strains showed high levels of susceptibility to the extract, except P. vibia, V. parvula and L. crispatus. Interestingly, all G. vaginalis clinical isolates and the G. vaginalis ATTC strain were the most susceptible to the extract, given their low MIC (1.0 - 2.0 mg/mL) and MBC (1.0 - 4.0 mg/mL) values, whereas, the Lactobacillus spp. clinical isolates and the L. crispatus ATCC strain were the least susceptible bacteria given their high MIC (32.0 mg/mL) and MBC (≥ 32.0 mg/mL) values. CONCLUSIONS.: In vitro experiments suggest that the extract possesses selective antimicrobial properties given its high activity against bacterial vaginosis-associated anaerobic bacteria and low activity against Lactobacillus species.

OBJETIVOS: Motivación para realizar el estudio: la vaginosis bacteriana es una infección bacteriana que afecta de forma frecuente a las mujeres en edad reproductiva. El tratamiento se basa en antimicrobianos sintéticos. Bixa orellana L. posee propiedades antimicrobianas y podría representar una potencial alternativa terapéutica no sintética. Principales hallazgos: los resultados in vitro sugieren que el extracto metanólico de las hojas de Bixa orellana L. posee potenciales propiedades antimicrobianas contra las bacterias asociadas a vaginosis bacteriana. Implicancias: identificar nuevas fuentes con potencial terapéutico, y promover la investigación, descubrimiento y caracterización de antimicrobianos no sintéticos. Describir la actividad antimicrobiana in vitro del extracto metanólico de las hojas de Bixa orellana L. contra las bacterias anaerobias asociadas a la vaginosis bacteriana y Lactobacillus spp. MATERIALES Y MÉTODOS: . Se incluyeron en el estudio ocho cepas de referencia ATCC; Gardnerella vaginalis, Prevotella bivia, Peptococcus niger, Peptostreptococcus anaerobius, Mobiluncus curtisii, Atopobium vaginae, Veillonella parvula y Lactobacillus crispatus, y 22 aislamientos clínicos; once aislados de Gardnerella vaginalis y once aislados de Lactobacillus. La susceptibilidad antimicrobiana se determinó mediante el método de difusión en agar. La concentración mínima inhibitoria (CMI) y la concentración bactericida mínima (CBM) fueron determinadas utilizando el método de dilución en agar y un método de dilución modificado, respectivamente. RESULTADOS: . Todas las cepas de referencia ATCC tuvieron un alto nivel de susceptibilidad al extracto, con excepción de P. vibia, V. parvula y L. crispatus. Interesantemente, los aislamientos clínicos de G. vaginalis y la cepa ATCC de G. vaginalis fueron los más susceptibles al extracto dados los bajos valores de CMI (1,0 - 2,0 mg/mL) y CBM (1,0 - 4,0 mg/mL), mientras que, los aislamientos clínicos de Lactobacillus spp. y la cepa ATCC de L. crispatus fueron los menos susceptibles debido a los altos valores de CMI (32,0 mg/mL) y CBM (≥ 32,0 mg/mL). CONCLUSIONES: . Los experimentos in vitro sugieren que el extracto posee propiedades antibacterianas selectivas dada su alta actividad contra bacterias anaerobias asociadas a vaginosis bacteriana y baja actividad contra especies de Lactobacillus.

Frequency of colistin resistance in Pseudomonas aeruginosa: first report from Peru. / Frecuencia de resistencia a la colistina en Pseudomonas aeruginosa: primer reporte en el Perú.

This study aimed to determine the frequency of colistin resistance in Pseudomonas aeruginosa isolates obtained from three healthcare facilities in Lima and cryopreserved at the Laboratorio de Resistencia Antimicrobianos e Inmunopatología of the Universidad Peruana Cayetano Heredia (UPCH). The colistin broth disk elution method was used for the phenotypic identification of colistin resistance. We detected the expression of the mcr-1 gene by using the phenotypic diffusion method with combined colistin and ethylenediaminetetraacetic acid (EDTA) disks; and polymerase chain reaction (PCR) was used for molecular identification of the gene. Of the 97 isolates, 7 (7.2%) were resistant to colistin; however, none carried the mcr-1 gene. This is the first report from Peru on clinical isolates of colistin-resistant Pseudomonas aeruginosa, which suggests the need for implementation of appropriate methodologies for the epidemiological surveillance of colistin-resistant pathogens.

El objetivo del estudio fue determinar la frecuencia de resistencia a la colistina en Pseudomonas aeruginosa provenientes de tres establecimientos de salud de Lima, criopreservados en el banco de cepas del Laboratorio de Resistencia a Antimicrobianos e Inmunopatología de la Universidad Peruana Cayetano Heredia (UPCH). El método de elución de discos de colistina en caldo fue empleado para la identificación fenotípica de la resistencia a la colistina; la detección de la expresión del gen mcr-1 se realizó mediante el método fenotípico de difusión de discos combinados de colistina y ácido etilendiaminotetraacético (EDTA) y la reacción en cadena de la polimerasa (PCR) para la identificación molecular del gen. De los 97 aislados estudiados, 7 (7,2%) fueron resistentes a la colistina y ninguno fue portador del gen mcr-1. Este estudio constituye el primer reporte en el Perú de aislados clínicos de Pseudomonas aeruginosa resistentes a la colistina, lo que implica la necesidad de implementar metodologías apropiadas para la vigilancia epidemiológica de patógenos resistentes a la colistina.

Antimicrobial Resistance Patterns and Dynamics of Extended-Spectrum ß-Lactamase-Producing Uropathogenic Escherichia coli in Cusco, Peru.

Urinary tract infections (UTIs) are a common human infection. Antibiotic resistance in extended-spectrum ß-lactamase (ESBL)-producing uropathogenic E. coli (UPEC) is a major therapeutic challenge due to limited treatment alternatives. The aim was to characterize the antimicrobial resistance (AMR) and dynamics of ESBL-producing UPEC isolates from UTI cases seen at a local hospital in Cusco, Peru. Ninety-nine isolates from respective patients were characterized against 18 different antibiotics. Latent class analysis (LCA) was used to evaluate the dynamics across the study time according to resistance patterns. The median age of patients was 51 years old, and nearly half were women. ESBL-producing UPEC isolates were slightly more frequent in outpatient services than emergency rooms, and there were higher resistance rates in males compared to females. Half of the ESBL producers were resistant to aminoglycosides and nitrofurantoin. Cefoxitin and fosfomycin resistance was 29.3% and 14.1%, respectively. Resistance to carbapenems was not observed. All isolates were multidrug-resistant bacteria, and 16.2% (16/99) were also classified as extensively drug-resistant bacteria. The resistance patterns varied across the study time and differed regarding sex and healthcare service. The study revealed high levels of AMR to commonly used antimicrobials and a dynamic circulation of ESBL-producing UPEC isolates with varying resistance patterns.

Frecuencia de resistencia a la colistina en Pseudomonas aeruginosa: primer reporte en el Perú

El objetivo del estudio fue determinar la frecuencia de resistencia a la colistina en Pseudomonas aeruginosa provenientes de tres establecimientos de salud de Lima, criopreservados en el banco de cepas del Laboratorio de Resistencia a Antimicrobianos e Inmunopatología de la Universidad Peruana Cayetano Heredia (UPCH). El método de elución de discos de colistina en caldo fue empleado para la identificación fenotípica de la resistencia a la colistina; la detección de la expresión del gen mcr-1 se realizó mediante el método fenotípico de difusión de discos combinados de colistina y ácido etilendiaminotetraacético (EDTA) y la reacción en cadena de la polimerasa (PCR) para la identificación molecular del gen. De los 97 aislados estudiados, 7 (7,2%) fueron resistentes a la colistina y ninguno fue portador del gen mcr-1. Este estudio constituye el primer reporte en el Perú de aislados clínicos de Pseudomonas aeruginosa resistentes a la colistina, lo que implica la necesidad de implementar metodologías apropiadas para la vigilancia epidemiológica de patógenos resistentes a la colistina.

This study aimed to determine the frequency of colistin resistance in Pseudomonas aeruginosa isolates obtained from three healthcare facilities in Lima and cryopreserved at the Laboratorio de Resistencia Antimicrobianos e Inmunopatología of the Universidad Peruana Cayetano Heredia (UPCH). The colistin broth disk elution method was used for the phenotypic identification of colistin resistance. We detected the expression of the mcr-1 gene by using the phenotypic diffusion method with combined colistin and ethylenediaminetetraacetic acid (EDTA) disks; and polymerase chain reaction (PCR) was used for molecular identification of the gene. Of the 97 isolates, 7 (7.2%) were resistant to colistin; however, none carried the mcr-1 gene. This is the first report from Peru on clinical isolates of colistin-resistant Pseudomonas aeruginosa, which suggests the need for implementation of appropriate methodologies for the epidemiological surveillance of colistin-resistant pathogens.

Draft Genome Sequence of a New Delhi Metallo-ß-Lactamase (NDM-1)-Producing Providencia stuartii Strain Isolated in Lima, Peru.

Providencia stuartii is an opportunistic pathogen of the Enterobacteriales order. Here, we report the 4,594,658-bp draft genome sequence of a New Delhi metallo-ß-lactamase (NDM-1)-producing Providencia stuartii strain that was isolated from an emergency patient in a private clinic in Lima, Peru.

Multidrug-resistant bacteria isolated from cell phones in five intensive care units: Exploratory dispersion analysis.

INTRODUCTION: Cell phones are susceptible to bacterial contamination. The aim of this study was to characterize the bacterial isolates and to explore their dispersion in five Intensive Care Units (ICUs) over the time. METHODS: We performed a secondary analysis of non-fermenting Gram-negative bacteria and Gram-positive cocci isolated from a 5-month observational cohort study developed among health care workers' cell phones in five ICUs. Cell phones were sampled using a swab every 15 days. Antimicrobial resistance was determined by the minimum inhibitory concentration method. We constructed resistance phenotypes to group the isolates according to species and antimicrobial resistance pattern to explore dispersion through time. RESULTS: A total of 35 P. aeruginosa, 16 Acinetobacter spp., 30 S. aureus and 26 Enterococcus spp. were isolated from 491 phone samples. Multidrug resistance was 2.9% for P. aeruginosa, 31.3% for Acinetobacter spp., 46.7% for S. aureus and 80.8% for Enterococcus spp. The resistance to methicillin in S. aureus and to vancomycin in Enterococcus spp. was 26.7% and 42.3%, respectively. We did not observe distribution patterns or clusters over the time for P. aeruginosa, Acinetobacter spp. and Enterococcus spp. isolates. All the S. aureus isolates grouped into eight phenotypes. Interestingly, we observed S. aureus isolates with the same phenotype in consecutive and separate sampling dates in the same cell phone. CONCLUSION: Cell phones are contaminated with highly harmful bacteria and potentially can maintain them for prolonged periods of time. These devices could be considered as a potential source of nosocomial infections in ICUs.

Draft Genome Sequence of the First New Delhi Metallo-ß-Lactamase (NDM-1)-Producing Escherichia coli Strain Isolated in Peru.

We present here the draft genome sequence of the first New Delhi metallo-ß-lactamase (NDM-1)-producing Escherichia coli strain, belonging to sequence type 155 (ST155), isolated in Peru. Assembly of this draft genome resulted in 5,061,184 bp, revealing a clinically significant resistome for ß-lactams, aminoglycosides, tetracyclines, phenicols, sulfonamides, trimethoprim, and fluoroquinolones.

Rapid detection of salmonellosis due to Salmonella enterica serovar Typhimurium in Peruvian commercially bred cavies, using indigenous wild bacteriophages.

INTRODUCTION: The salmonelloses are among the commonest, most widespread human zoonotic infections. They have generated international networks to attempt their control, since they cause a spectrum of ailments, ranging from inapparent carrier states to full-blown, severe, sometimes deadly diarrheal and systemic disease. Rapid diagnosis is needed for a number of reasons. The aim of this study was to standardize and validate a phage amplification test for the identification of salmonellosis to be applied to infections of Cavia porcellus. METHODS: Native bacteriophages were isolated from infected cavies and environmental residues from commercial cavy-breeding facilities. Salmonella enterica serovar Typhimurium ATCC 14028 was used to detect, isolate and propagate the bacteriophages, and to standardize a phage amplification assay to detect S. Typhimurium from rectal swabs of cavies. The phage amplification assay was tested using 2 antiviral agents, MgSO4·7H2O (MAS) and pomegranate rind extract (PRE) plus ferrous sulfate (PRE-FeSO4). RESULTS: The final assay format chosen used PRE-FeSO4 and allowed detection of S. Typhimurium in 90 min from culture, 5 h from clinical samples, with a limit of detection at 103 pfu; sensitivity was 98.2%, specificity 98%, negative predictive value (NPV) 96.1%, and positive predictive value (PPV) 99.1%. CONCLUSION: Bacteriophage amplification is therefore an appropriate, fast procedure for detection of this pathogen in clinical samples.

Characterisation of the first KPC-2-producing Klebsiella pneumoniae ST340 from Peru.

OBJECTIVES: The aim of this study was to characterise a KPC-carrying Klebsiella pneumoniae isolate from a Peruvian hospital setting. METHODS: The identity of the isolate was confirmed by amplification and sequencing of the 16S rRNA gene, and the antibiotic resistance profile was determined by disk diffusion and automated methods The sequence type (ST) and phylogenetic group were established by PCR. The presence of different ß-lactamase genes was determined, including blaMBL, blaKPC, blaCTX-M, blaSHV, blaOXA-1-like, blaOXA-2-like, blaOXA-5-like, blaOXA-48-like and blaTEM and up to six different plasmid-encoded AmpC genes as well as class 1 integrons. The conjugability of ß-lactam resistance was assessed by conjugation. RESULTS: The isolate was confirmed to be K. pneumoniae classified as belonging to the KpI phylogenetic group within ST340, which belongs to the high-risk clonal complex 258 (CC258). The isolate was resistant to all ß-lactam agents tested, with only the presence of a non-conjugative blaKPC-2 gene being detected and carried in a non-classical genetic structure. CONCLUSIONS: This is the first description of a member of CC258 and of a blaKPC-2 gene in Peru. Intensive surveillance is needed to determine the relevance of both in this area.

Caracterización fenotípica y molecular de Escherichia coli productoras de Beta-Lactamasas de espectro extendido en pacientes ambulatorios de Lima, Perú / Phenotypic and molecular characterization of extended spectrum beta-lactamase producing strains of Escherichia coli in ambulatory patients in Lima, Peru

Objetivos: Determinar las características fenotípicas y genotípicas de las β-lactamasas de espectro extendido (BLEE) en E. coli aislados de cultivos de orina de pacientes de la comunidad en un laboratorio privado de la ciudad de Lima, Perú. Material y métodos: Se evaluaron 53 aislamientos de E. coli por dos métodos fenotípicos: Jarlier y CLSI, el perfil de susceptibilidad se realizó mediante disco difusión y la caracterización genotípica mediante PCR para los genes blaCTX-M, blaTEM y blaSHV. Resultados: Los 53 aislamientos productores de BLEE representaron el 16,30% del total de aislados de E. coli, afectando principalmente a mujeres mayores de 65 años. El perfil de susceptibilidad evidenció alta resistencia a AMP,CEF,CRO(100%), LEV(87%), NOR(92%), CIP y NAL(94%), CXM y CTX(96%),SXT(70%), ATM(75%) y TOB (85%); asimismo elevada sensibilidad a NIT e IPM(100%), AMK(91%) y FOF(73,6%). El tipo de gen bla más frecuente fue blaCTX-M (55%), seguido por la coexistencia blaCTX-M+TEM (24%), blaTEM (13%) y blaSHV (6%). Conclusiones: La frecuencia de E. coli productores de BLEE fue de 16,3%; siendo el gen tipo blaCTX-M el más frecuente, información valiosa para orientar la terapia antimicrobiana empírica.

Objective: To determine the phenotypic and genotypic features of extended spectrum beta-lactamase (ESBL) producing strains of Escherichia coli isolated from urine samples of patients attending outpatient services in a private laboratory in Lima, Peru. Methods: 53 E. coli isolates were evaluated using two phenotypic methods: Jarlier and CLSI, the susceptibility profile was performed using the disk diffusion method and the genotypic features were analyzed using PCR for detecting blaCTX-M, blaTEM y blaSHV genes. Results: The 53 ESBL producing strains of E. coli accounted for 16,30% of all E. coli isolates affecting mostly women older than 65 years. High resistant profile to AMP, CEF, CRO (100%), LEV (87%), NOR (92%), CIP, NAL (94%), CXM, CTX (96%), SXT (70%), ATM (75%) and TOB (85%) was observed. High susceptibility to NIT, IPM (100%), AMK (91%) and FOF (73.6%) was observed. The most frequent bla gen was blaCTX-M (55%), followed by blaCTX-M+TEM (24%), blaTEM (13%) and blaSHV (6%). Conclusions: The rate of ESBL producing strains of E. coli was 16.3% and the blaCTX-M gen was the most common gene type. These results provide valuable information for starting empiric antibiotic therapy in this setting.

Extended-spectrum ß-lactamase-producing Enterobacteriaceae in cell phones of health care workers from Peruvian pediatric and neonatal intensive care units.

BACKGROUND: Health care workers (HCWs) use their mobile phones during working hours or medical care. There is evidence that the instruments are colonized with pathogenic microorganisms. Here, we describe levels of Enterobacteriaceae contamination (EC) in cell phones and the risk factors associated with EC in Peruvian intensive care units (ICUs). METHODS: This was a 5-month cohort study among 114 HCWs of 3 pediatric and 2 neonatology ICUs from 3 Peruvian hospitals. A baseline survey collected data on risk factors associated with EC. Swabs were collected from HCWs' phones every other week. RESULTS: Three-quarters of HCWs never decontaminated their phones, and 47% reported using the phones in the ICU >5 times while working. EC was frequent across samplings and sites and was substantially higher in subjects with longer follow-up. Potential risk factors identified did not have strong associations with positive samples (relative risk, 0.7-1.5), regardless of significance. Half of the phones were colonized with an Enterobacteriaceae at least once during the 4 samplings attained on average during the study period. Half of the isolates were multidrug resistant (MDR), and 33% were extended-spectrum ß-lactamase producers. CONCLUSIONS: EC on HCWs' phones was frequent and apparently randomly distributed through the hospitals without clear clustering or strongly associated risk factors for having a positive sample. Based on the level of EC, phones may be considered as potential bacterial reservoirs of MDR and ESBL bacteria.

[Phagotherapy faced with Staphylococcus aureus methicilin resistant infections in mice]. / Fagoterapia frente a infecciones por Staphylococcus aureus meticilino resistente en ratones.

OBJECTIVES: To assess the bacteriophage activity in localized and systemic infections caused by Staphylococcus aureus resistant to methicilin (MRSA). MATERIALS AND METHODS: An experimental study was performed in 45 mice of the Balb/c strain divided in nine groups of five individuals. Ten naive bacteriophages were isolated through clinical samples and hospital effluents. Lytic capacity and spectrum activity was evaluated on the basis of which six phages were selected for phagotherapy trials. Additionally, a commercial bacteriophage was used. The phagotherapy was evaluated through prophylaxis, and therapy of localized and systemic infections caused by MRSA by subcutaneous and intravenous inoculation, respectively. The effectiveness of three therapeutic schemes was tested: monotherapy, phage cocktail in multiple doses and phage cocktail in a single dose. The therapeutic activity of the phages was also compared with vancomycin and clindamycin. RESULTS: The phage cocktail and the diverse dose therapy were effective in preventing and controlling MRSA localized infections; its activity was similar to the vancomycin and clindamycin activity. The single dose phage cocktail failed to control localized infection and phagotherapy was not effective in systemic infections. CONCLUSIONS: Phagotherapy could be a viable alternative for infections caused by MRSA. Further studies that assess related aspects to phages and patient safety are required.

Serotipos y susceptibilidad antimicrobiana de Shigella aisladas en un instituto de salud pediátrico de Lima, Perú entre enero y julio 2013 / Serotypes and antimicrobial susceptibility of Shigella isolated in Pediatric Health Institute in Lima, Peru from January to July 2013

Objetivos: Determinar la frecuencia de serogrupos y serotipos y el perfil de susceptibilidad antimicrobiana de Shigella sp., aisladas en un instituto pediátrico de Lima, Perú. Material y métodos: Se evaluaron 85 aislamientos de Shigella sp., identificados bioquímicamente y serológicamente a nivel de serogrupo y serotipo por el método de aglutinación en lámina. Los patrones de resistencia antibiótica se determinaron mediante el método de disco difusión en agar. Resultados: De los 85 aislamientos de Shigella sp., 53 (62,3%) correspondieron al serogrupo B (Shigella flexneri), 28 (32,9%) al grupo D (Shigella sonnei) y 4 (4,8%) al grupo C (Shigella boydii), ningún aislamiento correspondió al grupo A (Shigella dysenteriae). Respecto a los serotipos, en el grupo B, fueron 46% 1b, 36% 2a y 18% variante Y; en el grupo C fue C4 y en el grupo D todos fueron Fase I. La evaluación del perfil de susceptibilidad mostró que el 100% de las cepas fueron sensibles a aztreonam, ácido nalidíxico y ciprofloxacina; entre 80 y 90% fueron resistentes a trimetoprim-sulfametoxazol, ampicilina y tetraciclina. Conclusiones: El serogrupo más frecuente fue Shigella flexneri, no se reportó Shigella dysenteriae. Existe elevado nivel de resistencia a Sulfametoxazole/trimetoprim, ampicilina y tetraciclina. (AU)

Objectives: To determine the frequency of serogroups and serotypes and antimicrobial susceptibility profile of Shigella sp., isolated in a pediatric institute from Lima, Peru. Methods: Observational study conducted in 85 isolates of Shigella sp., serologically identified to serogroup and serotype by slide agglutination method. The antibiotic resistance patterns were determined by disk diffusion in agar. Results: 53 (62.3%) were serogroup B (Shigella flexneri), 28 (32.9%) group D (Shigella sonnei) and 4 (4.8%) group C (Shigella boydii) , no bacteria group A (Shigella dysenteriae) was isolated. Respect to serotypes, in group B were 46% 1b, 36% 2a y 18% Y variant; in group C was C4 and in group D all were phase I. Evaluation of susceptibility profile showed that 100% of the strains were susceptible to aztreonam, nalidixic acid and ciprofloxacin; between 80 and 90% were resistant to trimethoprim-sulfamethoxazole, ampicillin and tetracycline. Conclusions: The most common serogroup was Shigella flexneri. Shigella dysenteriae was not reported. There is a high level of resistance sulfamethoxazole / trimethoprim, ampicillin and tetracycline. (AU)

Fagoterapia frente a infecciones por Staphylococcus aureus meticilino resistente en ratones / Phagotherapy faced with Staphylococcus aureus methicilin resistant infections in mice

Objetivos: Evaluar la actividad de los bacteriófagos frente a infecciones localizadas y sistémicas producidas por Staphylococcus aureus resistente a meticilina (MRSA) Materiales y métodos. Se realizó un estudio de tipo experimental en 45 ratones de la cepa Balb/c divididos en nueve grupos de cinco individuos. Se aislaron diez bacteriófagos nativos a partir de muestras clínicas y efluentes hospitalarios, se evaluó su capacidad lítica y su espectro de actividad, en base a lo cual se seleccionaron seis fagos para los ensayos de fagoterapia. Adicionalmente, se empleó un bacteriófago de origen comercial. La fagoterapia fue evaluada mediante profilaxis y terapia de infecciones localizadas y sistémicas causadas por la inoculación de MRSA por vía subcutánea y endovenosa respectivamente. Se probó la efectividad de tres esquemas terapéuticos: monoterapia, cóctel de fagos en múltiples dosis y de cóctel de fagos en una sola dosis. También se comparó la actividad terapéutica de los fagos frente a vancomicina y clindamicina. Resultados. El cóctel de fagos y la terapia a diversas dosis fueron efectivos para prevenir y controlar infecciones localizadas por MRSA, su actividad fue similar a la de vancomicina y clindamicina. La dosis única del cóctel de fagos no logró controlar la infección localizada; asimismo, la fagoterapia no resultó efectiva en infecciones sistémicas. Conclusiones. La fagoterapia se proyecta como una alternativa viable frente a infecciones causadas por MRSA. Se requieren estudios que evalúen aspectos relacionados con la inocuidad de los fagos frente al paciente.

Objectives: To assess the bacteriophage activity in localized and systemic infections caused by Staphylococcus aureus resistant to methicilin (MRSA). Materials and methods. An experimental study was performed in 45 mice of the Balb/c strain divided in nine groups of five individuals. Ten naive bacteriophages were isolated through clinical samples and hospital effluents. Lytic capacity and spectrum activity was evaluated on the basis of which six phages were selected for phagotherapy trials. Additionally, a commercial bacteriophage was used. The phagotherapy was evaluated through prophylaxis, and therapy of localized and systemic infections caused by MRSA by subcutaneous and intravenous inoculation, respectively. The effectiveness of three therapeutic schemes was tested: monotherapy, phage cocktail in multiple doses and phage cocktail in a single dose. The therapeutic activity of the phages was also compared with vancomycin and clindamycin. Results. The phage cocktail and the diverse dose therapy were effective in preventing and controlling MRSA localized infections; its activity was similar to the vancomycin and clindamycin activity. The single dose phage cocktail failed to control localized infection and phagotherapy was not effective in systemic infections. Conclusions. Phagotherapy could be a viable alternative for infections caused by MRSA. Further studies that assess related aspects to phages and patient safety are required.

[Comparison of four phenotypic methods to detect extended-spectrum betalactamases]. / Comparación de cuatro métodos fenotípicos para la detección de beta-lactamasas de espectro extendido.

OBJECTIVE: To compare the efficacy of four phenotypic methods for the identification of strains producing extended-spectrum ß-lactamases isolated from urine cultures. MATERIALS AND METHODS: Comparative cross-sectional study. 147 strains isolated from positive urine cultures for Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis between January and February 2009 in the National Institute of Health of Children underwent a screening test, those which resulted positive were processed for confirmatory testing through the four phenotypic methods evaluated. RESULTS: Out of the 147 strains, 43 (29.3%) were suspicious in the screening tests. Using the method described by the Clinical and Laboratory Standards Institute (CLSI) as a reference standard for this study, 27 strains (62.8%) were positive, with similar results using Jarlier's method. On the other hand, using Hodge's and tridimensional methods 23 (53.5%) of samples were positive. The evaluation of the confirmation methods in comparison to the one described by CLSI, showed a sensitivity and specificity of 100% for Jarlier's method, on the other hand, for Hodge's and tridimensional methods we found a sensitivity of 85.2% and 100%, respectively. CONCLUSIONS: All the evaluated methods showed a high efficacy, without significant differences, so they could all be used according to the available facilities of each clinical laboratory. Nevertheless, due to its advantages besides the technical aspect, like costs, easiness and feasibility of its application, we recommend the use of Jarlier's method.

Comparación de cuatro métodos fenotípicos para la detección de beta-lactamasas de espectro extendido / Comparison of four phenotypic methods to detect extended-spectrum betalactamases

Objetivo. Comparar la eficacia de cuatro métodos fenotípicos para la identificación de cepas productoras de lactamasas de espectro extendido aisladas de urocultivos. Materiales y métodos. Estudio comparativo de corte transversal. Se analizó 147 cepas aisladas de urocultivos positivos para Escherichia coli, Klebsiella pneumoniae y Proteus mirabilis entre los meses de enero y febrero de 2009 en el Instituto Nacional de Salud del Nino, las cuales fueron sometidas a una prueba de tamizaje, en aquellas que resultaron positivas se realizo pruebas confirmatorias mediante los cuatro métodos fenotípicos evaluados. Resultados. De las 147 cepas, 43 (29,3 por ciento) resultaron sospechosas en las pruebas de tamizaje. Con el método descrito por Clinical and Laboratory Standards Institute (CLSI) usado como patron de oro para el estudio, resultaron 27 positivas (62,8 por ciento), resultado similar a lo encontrado con el método de Jarlier. Por otro lado, con los métodos de Hodge y el tridimensional, 23 (53,5 por ciento) resultaron positivas. La evaluación de los métodos confirmatorios frente al descrito por CLSI, mostro una sensibilidad y especificidad de 100 por ciento para el método de Jarlier; en el caso de los métodos de Hodge y método tridimensional se encontró una sensibilidad y especificidad de 85,2 por ciento y 100 por ciento, respectivamente. Conclusiones. Los métodos evaluados mostraron en todos los casos una alta eficacia, sin presentar diferencias significativas, por lo que podrían utilizarse según las facilidades del laboratorio clínico involucrado; sin embargo dada las ventajas no técnicas, como costos, facilidad y factibilidad de su aplicación, se recomienda el empleo del método de Jarlier.

Objective. To compare the efficacy of four phenotypic methods for the identification of strains producing extended-spectrum Beta-lactamases isolated from urine cultures. Materials and methods. Comparative cross-sectional study. 147 strains isolated from positive urine cultures for Escherichia coli, Klebsiella pneumoniae and Proteus mirabilis between January and February 2009 in the National Institute of Health of Children underwent a screening test, those which resulted positive were processed for confirmatory testing through the four phenotypic methods evaluated. Results. Out of the 147 strains, 43 (29.3 percent) were suspicious in the screening tests. Using the method described by the Clinical and Laboratory Standards Institute (CLSI) as a reference standard for this study, 27 strains (62.8 percent) were positive, with similar results using Jarlier's method. On the other hand, using Hodge's and tridimensional methods 23 (53.5 percent) of samples were positive. The evaluation of the confirmation methods in comparison to the one described by CLSI, showed a sensitivity and specificity of 100 percent for Jarlier's method, on the other hand, for Hodge's and tridimensional methods we found a sensitivity of 85.2 percent and 100 percent, respectively. Conclusions. All the evaluated methods showed a high efficacy, without significant differences, so they could all be used according to the available facilities of each clinical laboratory. Nevertheless, due to its advantages besides the technical aspect, like costs, easiness and feasibility of its application, we recommend the use of Jarlier's method.

Staphylococcus aureus resistente a meticilina adquirido en la comunidad aislados en tres hospitales de Lima-Perú / Methicillin resistant Staphylococcus aureus isolates acquired in the community in three hospitals in Lima, Peru

Staphylococcus aureus es un importante patógeno involucrado en una serie de infecciones e intoxicaciones, presenta múltiples factores de virulencia y su impacto se incrementa por su notable resistencia a los antimicrobianos. Objetivo: Determinar la frecuencia de Staphylococcus aureus meticilino resistente adquiridos en la comunidad, en hospitales de Lima- Perú. Material y métodos: Se realizó un estudio descriptivo multicéntrico. La resistencia a meticilina se determinó por el método Oxacillin Agar Screen. El origen de la cepa fue determinado mediante los criterios de los CDC; la Leucocidina de Panton Valentine fue identificada por métodos moleculares. Resultados: Se aislaron 276 cepas de Staphylococcus aureus, 160 fueron resistentes a meticilina (58 por ciento), 9 de ellas fueron identificadas como adquiridas en la comunidad (5,6 por ciento). La PVL fue identificada en 25 cepas (9,1 por ciento), 14 fueron MSSA y 11 MRSA, de éstas últimas solo 4 fueron MRSAcom, 7 fueron MRSAhosp (p menor que 0,001). Conclusiones: El estudio revela niveles elevados de resistencia a meticilina, pero niveles bajos de MRSAcom. En nuestro medio la presencia de PVL no constituiría un marcador para la identificación de los MRSAcom.

Background: Staphylococcus aureus is an important pathogen involved in a series of infections and toxin mediated syndromes, has many virulence factors and its impact increases its resistance to antimicrobial agents. Objectives: To determine the frequency of community acquired methicillin resistant Staphylococcus aureus among hospitals in Lima -Peru. Material and Methods: We performed a prospective multicenter study. The resistance to methicillin was determined by the Oxacillin Agar Screen method. The origin of the strain was determined using CDC criteria, the Panton Valentine Leucocidin was identified by molecular methods. Results: We isolated 276 strains of Staphylococcus aureus, 160 were resistant to methicillin (58 per cent). 9 strains were identified as community acquired MRSA (5.6 per cent). The Panton Valentine Leucocidin was identified in 25 strains (9.1 per cent), 14 were MSSA and 11 MRSA, only 4 of the latter were CA MRSA, 7 were HA MRSA (p less than 0.001). Conclusions: The study showed frequencys of methicillin resistance, but low CA MRSA. In our environment the presence of PVL would not be a marker for the identification of CA MRSA.