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The aim of the study was to determine the frequency of six genes encoding serine protease autotransporter proteins Enterobacteriaceae (SPATE) in diffusely adherent Escherichia coli (DAEC) isolates from children with (WD, n=63) and without diarrhea (WOD, n=41) from Lima, Peru. WOD were considered a control group. For the detection of the genes, 2 multiple PCRs were standardized: triple A (sigA, pet, espP) and triple B (sat, pic, espC). In both groups, the most frequent SPATE gene was Sat (39.7% of WD and 41.5% of WOD), followed by spP (20.6% and 9.7% in WD and WOD respectively). The other genes were detected in proportions lower than 10.0%, in the following order of frequency: pet, sigA, espC and pic, without significant differences between the groups. It was concluded that Sat is the most frequent SPATE in DAEC and that these strains may possess SPATE genes regardless of whether they are isolated in WD or WOD.

The present report is the original description of bla TEM-176. The mechanisms of resistance to antimicrobial agents were determined in an enterotoxigenic Escherichia coli, determining the susceptibility to 22 antimicrobials classified in 15 different groups by agar diffusion and establishing the phylogenetic group, mechanisms of resistance and presence of Class 1 and 2 integrons. Integrons and β-lactam resistance genes were sequenced. The isolate, belonging to phylogenetic group A, showed the presence of resistance or diminished susceptibility to a ampicillin, amoxicillin plus clavulanic acid, nalidíxic acid, ciprofloxacin, streptomycin, kanamycin, tetracycline, trimethoprim, sulfisoxazole, cotrimoxazole, azithromycin and nitrofurantoin, carrying bla TEM, aadA1/2, aphA1, sul3, tet(A) and a Class 2 integron containing a dfrA1 gene. Quinolone resistance was related to the substitution Ser83Ala. The TEM sequencing showed the presence of the new substitution Ala222Val, which led to the description of the new β-lactamase bla TEM-176.

Introduction: We evaluated the presence of sIgA in saliva, versus Escherichia coli secreted proteins (Esp) related to the type III secretion system (T3SS), and its semi-quantitative concentration in children under 2 years-old (no longer breastfed) who were previously colonized or infected with enteropathogenic E. coli (EPEC)., Methods: We analyzed the presence of sIgA in 40 children, who previously had positive cultures for EPEC associated (n=17) or not associated (n=23) with diarrhea, using the Western Blot technique versus E. coli secreted proteins: EspABCD. A semi-quantitative measurement of the reaction for each protein was made by its density peaks (OD)., Results: We found sIgA versus all or some EspABCD proteins in both groups. However, the ill patients had higher concentrations of these antibodies than colonized patients., Discussion: The presence of sIgA in saliva could reflect an intestinal immune response and their levels could be related to a greater exposure and/or bacterial load., (Copyright © 2019 Elsevier España, S.L.U. and Sociedad Española de Enfermedades Infecciosas y Microbiología Clínica. All rights reserved.)

The mechanisms of resistance to nitrofurans from 18 meat samples with Salmonella enterica (chicken: 15; beef: 2; pork: 1) collected in Lima (Peru) were analyzed. The isolates were serotyped and the susceptibility levels to furazolidone and nitrofurantoin [with and without the efflux pump inhibitor Phenyl-Arginine- β-naphthylamide (PAβN)], the presence of mutations in the snrA and cnr genes and the transferability of resistance by conjugation were established. Fifteen samples with S. infantis (13 from chicken samples), 2 with S. enteritidis and 1 with S. anatum were identified. All isolates except the S. anatum were resistant to both nitrofurans showing MICs (minimum inhibitory concentration) of furazolidone and nitrofurantoin of 32-64 μg/mL and 128-256 μg/mL, respectively. The addition of PAßN had no effect on the MIC levels. All nitrofuran-resistant isolates showed amino acid codon alterations at both snrA and cnr (S. infantis: snrA STOP-151; cnr STOP-137; S. enteritidis: snrA STOP-180; cnr STOP-179). No transferable mecha nisms of nitrofuran resistance were detected.

Antibiotic resistance is a major global problem. The objective of this study was to determine antibiotic resistance in commensal strains isolated from healthy children from rural communities of Moyobamba and Urubamba in Peru. This cohort study identified 179 commensal E. coli strains from 93 children, followed for six months. Thirteen antibiotics were analyzed by diffusion disk. The highest rates of resistance were for cotrimoxazole (49.1%), ampicillin (48.0%), and nalidixic acid (31.8%). An 11.6% increase in resistance was found for nalidixic acid and 6.4% for cotrimoxazole in this period; while 34.0% of the isolates were multidrug-resistant. This study supports previous findings of multidrug resistance in commensal strains in rural communities and highlights the increased rates of resistance over time. We recommend studies in larger populations with a longer follow-up.

To describe the frequency and severity of intraventricular hemorrhage and periventricular leukomalacia in low birth-weight neonates in three hospitals in Lima, Peru, 385 newborn babies weighing under 2,000 g at birth were evaluated between May 2012 and July 2014. Brain ultrasounds were obtained at 40 weeks' gestation, 3-5 days of life, and 3-4 weeks of life. Intraventricular hemorrhage occurred in 19.2% of neonates weighing under 1,500 g and was severe (grade III or with periventricular hemorrhagic infarction) in 9.6% of neonates under 1,500 g. Mortality in infants with intraventricular hemorrhage was 47.1%, while periventricular leukomalacia was found in 5.4% of neonates 1,500 g and under; both diagnoses were more frequent in lower-weight babies. The frequency of intraventricular hemorrhage is similar to that reported in other countries; however, severity and mortality are greater.

Objective.: The objective of this study was to describe the presence of Enterobacteriaceae in meat samples collected in traditional markets of Lima and to establish the levels of antimicrobial resistance and the presence of extended spectrum beta-lactamases (BLEE) and AmpC in Escherichia coli., Materials and Methods.: A total of 138 meat samples, 64 (46.4%) chicken, 44 (31.9%) beef and 30 (21.7%) pork were collected. The isolated bacteria belonged to 17 different genera and, specifically, 14 were classified as Enterobacteriaceae. Sensitivity to ten antimicrobial agents was analyzed using the Kirby-Bauer disc diffusion method, BLEE and AmpC were determined by double disc and imipenem-ceftazidime induction tests, respectively., Results.: Antimicrobial resistance levels were high against trimethoprim-sulfamethoxazole, ampicillin, tetracycline, nalidixic acid, ciprofloxacin and chloramphenicol. There are significant differences in antibiotic resistance levels depending on the type of meat (chicken, beef and pork) (p <0.05). Multiple drug resistance (MDR) levels were particularly high in chicken and pork (98.2% and 86.4%, respectively). In addition, the presence of BLEE in Escherichia coli isolated from chicken meat was 59.4%., Conclusions.: Multiple drug resistance levels were high compared to antibiotics frequently used in humans; chicken and beef are highlighted as potential reservoirs of BLEE and pAmpC-producing Escherichia coli, respectively.

Objective.: To determine the acceptability of purified fish protein (FPi) in cereal and pseudocereal-based drinks., Materials and Methods.: The study was carried out at Centro de Desarrollo Integral Familiar "Año Nuevo" belonging to the INABIF Program in the district of Comas in Lima. FPi (0-4 g per serving) was administered daily for 3 months mixed with cereal and pseudocereal-based drinks. The acceptability of FPi was measured by direct weighing in children under 6 years of age and by the hedonic facial scale in school-age children., Results.: One hundred and fifty-eight (158) participants were enrolled and 4,211 breakfasts were served. The caloric intake of the cereal and pseudocereal drinks, after adding the FPi, ranged between 117 and 140 Kcal, and protein intake ranged between 3.4 g and 5.4 g per serving. The acceptability of drinks supplemented with FPi in children older than 6 years of age (the 2 highest scores together) ranged from 55% to 74% for oats, around 47% for quinoa and 40% to 58% for "kiwicha" (amaranth). Whereas 90% of children under 6 consumed more than 70% of the drinks offered., Conclusions.: We found a good acceptability of FPi in cereal and pseudocereal beverages in children under 6 years of age. FPi is an alternative to improve the consumption of animal protein in the nutrition of children, especially in pre-schoolers.

Objectives.: To describe the clinical characteristics, antibiotic resistance, and distribution of serotypes of bacterial strains that cause invasive pneumococcal disease (IPD) in adults., Materials and Methods.: Case series. Pneumococcal strains were isolated from 2009 to 2011 from hospitalized adult patients with IPD in five hospitals and two laboratories located in Lima., Results.: The analysis of data from 43 patients with IPD indicated that 58.2% were older than 60 years. The most common complications were pneumonia (39.5%), meningitis (30.2%), bacteremia (13.9%), peritonitis (11.6%), and septic arthritis (4.8%). The mortality rate was 28.9%, and 72.7% of cases involved patients older than 60 years. The pneumococcal strains were resistant to the following antibiotics: penicillin, 0% and 30.8% in non-meningitis and meningitis strains, respectively; ceftriaxone, 4.5% and 16.7% in non-meningitis and meningitis strains, respectively; trimethoprim/sulfamethoxazole, 69.0%; and erythromycin, 35.7%. The most common serotypes were 19F, 23F, 6B, 14, and 6C. The percentage of vaccine strains was 44.2% for the 7-valent conjugate pneumococcal vaccine (PCV7) and PCV10, 51.2% for PCV13, and 60.4% for the 23-valent polysaccharide vaccine (PPV23)., Conclusions.: Pneumococcus is an important pathogen in adults, particularly in older adults, owing to its high mortality rate.

The empirical treatment of urinary infections is complicated by the presence of multiresistance and resistance to extendedspectrum beta-lactamases (ESBLs). The objective of this study was to describe the patterns of antibiotic resistance of Escherichia coli strains isolated from urine cultures and the clinical-epidemiological factors associated with the presence of ESBLs in a pediatric and an adult group. A total of 353 strains were collected from the Emergency and Hospitalization Sector of the Cayetano Heredia Hospital over 14 months, and 45.9% of the isolated strains were multiresistant. The rate of resistance to ESBLs in the pediatric and adult population was 16.3% and 31.1%, respectively, and 63.6% of the resistant strains were isolated from outpatients. The presence of ESBLs was associated with hospitalization in pediatrics, use of diapers, and the presence of neurogenic bladder in adults. These factors should be considered in selection of an antibiotic treatment. Moreover, epidemiological reporting programs and models should be implemented for reduction of risk factors.

The objective of this review was to provide guidelines regarding the diagnosis, treatment, and prevention of the most common infectious diseases in children during natural disasters, including floods such as those that occurred in the summer of 2017 in Peru. This instrument should be used by physicians and health personnel working in primary health care. The management of diarrheal, respiratory, skin, and eye diseases, vector-transmitted diseases such as dengue, chikungunya, and Zika, and zoonoses such as leptospirosis is discussed. Emphasis is placed on the essential aspects of health education for the population, particularly preventive health, including the use of potable water, breastfeeding, immunization, hand washing, and vector control.

Objectives: To determine the prevalence of purchase of antibiotics without prescription and antibiotic recommendation for its use in children <5 years of age in private pharmacies in peri-urban areas of Lima., Materials and Methods: An anonymous population survey was conducted among adults who bought an antibiotic for a child <5 years of age from a private pharmacy. Fictitious cases were applied in a subgroup of pharmacies to evaluate antibiotic recommendation practices., Results: The prevalence of purchase of antibiotics without a prescription was 13%, including 1.7% that were chosen by consumers and 11.3% that were recommended by pharmacists. A total of 66.7% of the medications purchased to treat colds were antibiotics, 56.9% of which were prescribed. 64.4% of medications purchased to treat bronchospasm, 96.4% of the medications to treat watery diarrhea and 90.9% of the medications to treat pharyngitis were antibiotics. Amoxicillin (51.5%) and trimethoprim-sulfamethoxazole (20.6%) were the most commonly purchased antibiotics. A pharmacist suggested an antibiotic in 60% of the fictitious common cold cases, 76% of the bronchospasm cases, 44% of the watery diarrhea cases, and 76% of the dysentery cases., Conclusions: The purchase of antibiotics for children <5 years of age in private pharmacies primarily occurs with a medical prescription. The pharmacist commonly recommended an antibiotic in cases lacking a prescription. Measures are needed to reduce the inappropriate use of antibiotics at medical facilities and in pharmacies.

The objective of this study was to analyze the use of lumbar punctures (LP) in the evaluation of late-onset neonatal sepsis. It is recommended to perform an LP as part of the evaluation of late-onset sepsis. We used a cohort of 414 newborns with a birth weight <2000g in three hospitals in Lima. A LP was performed in 45/214 (21.0%) of sepsis evaluations and in 13/48 (27.1%) of culture-proven sepsis. Meningitis was diagnosed in 8/214 (3.7%) of the episodes and 8/45 (17.5%) of the evaluations that included an LP. The duration of treatment of the sepsis episodes without a LP and the episodes with a negative LP was similar, and shorter than the episodes with a positive LP. The use of LP in the evaluation of late-onset sepsis is low and can result in undiagnosed and undertreated meningitis. The use of LP in the evaluation of neonatal sepsis must be encouraged in the neonatal units.

Objectives: To determine the pattern of antibiotic susceptibility of isolated Streptococcus pneumoniae strains of healthy nasopharyngeal carriers younger than 2 years in seven regions of Peru., Materials and Methods: Between 2007 and 2009, nasopharyngeal swab samples were collected among 2123 healthy children aged 2-24 months in growth and development medical practices (CRED) and vaccination offices of hospitals and health centers in Lima, Piura, Cusco, Abancay, Arequipa, Huancayo, and Iquitos. The resistance to ten antibiotics through disk diffusion sensitivity testing of isolated pneumococcus strains was determined., Results: 572 strains were isolated. High rates of resistance to co-trimoxazole (58%), penicillin (52.2% non-sensitive); tetracycline (29,1%); azithromycin (28,9%), and erythromycin (26,3%). Resistance to chloramphenicol was low (8.8%). Multiresistance was found at 29.5%. Resistance to azithromycin and penicillin was different in all seven regions (p<0,05), the highest percentage of non-sensitive strains being found in Arequipa (63,6%), whereas the lowest percentage was found in Cusco (23.4%)., Conclusions: High levels of resistance found to penicillin, co-trimoxasole and macrolides in isolated pneumococcus strains of healthy carriers in all studied regions, and their association to a previous use of antibiotics, represent a significant public health problem in our country. This emphasizes the need to implement nationwide strategies to reduce the irrational use of antibiotics, especially among children. It is necessary to complement data of resistance to penicillin with the determination of minimal inhibitory concentration to make proper therapeutic recommendations.

Objectives: To describe physicians' preferences of antibiotic use in children less than 5 years in health centers of primary level in three periurban districts in Lima, Perú., Material and Methods: A structured survey was applied to 218 general practitioners from three health networks of Lima. The survey included six typical clinical cases in children under 5 years with questions about antibiotic use: the cases were common cold, pharyngitis, pneumonia, bronchial obstructive syndrome, watery diarrhea and dysentery., Results: 81.6% of the physicians responded that more than a quarter of the patients they attended were children under five years. 15.6% of the general physicians would use an antibiotic for common cold treatment. For dysentery treatment 90.4% would use antibiotics, the frequently used were Trimethoprim-Sulfamethoxazole (TMP-SMX) and Furazolidone. 86.2% of physicians would recommend an antibiotic for pharyngitis. In a watery diarrhea case 32.7% of the doctors would use. In the case of bronchospasm, 73% of the doctors would recommend an antibiotic. 96.3% would recommend antibiotics for pneumonia. The perception of the degree of mother's satisfaction increased the risk of inappropriate prescription of antibiotics OR: 1.6, p=0.031, 95% CI: 1.1-2.6)., Conclusions: There is tendency to overuse antibiotics for diagnoses such as pharyngitis and bronchospasm, as well as in cases of watery diarrhea and common cold, the reason could be that a large number of children under five years are treated by general practitioners without training in pediatric care.

The main aim of this study was to establish the resistance levels to antimicrobial agents, in 222 non-pathogenic E. coli strains of fecal origin in Peru. The proportion of resistance found to the evaluated antimicrobials was ampicillin (62.6%), cotrimoxazole (48,6%), tetracycline (43,0%) and chloramphenicol (15,8%). We emphasize the high resistance levels found for quinolones: 32% for nalidixic acid (NAL) and 12% for ciprofloxacin (CIP). These high levels of quinoloneresistance in non-pathogenic strains isolated from children in this age group highlight the extensive use and the impact of the intake of this kind of antimicrobials in the community, showing the potential risk of the loss of their utility in the area.

Objectives: To determine the carriage rate and serotype distribution of Streptococcus pneumoniae in the nasopharynx of healthy children younger than 2 years prior to the universal use of the pneumococcal conjugate vaccines in Peru., Materials and Methods: Between 2007 and 2009 we collected nasopharyngeal swab samples from 2,123 healthy children aged 2 to 24 months in the vaccination and healthy children consultation offices of pediatric hospitals and health centers in 7 cities in Peru: on the coast (Lima, Piura), highlands (Cusco, Abancay, Arequipa and Huancayo) and amazon basin (Iquitos). The pneumococcal strains were isolated and identified at the central laboratory of the project in Lima, and serotyped by Quellung reaction in the pneumococcal reference laboratory at the Center for Diseases Control and Prevention (CDC)., Results: We found 27% (573/2123) of pneumococcal nasopharyngeal healthy carrier children. Among the 526 analyzed strains, we found 42 serotypes; the most common were: 19F (18.1%), 6B (14.3%); 23F (8.9%) and 14 (6.5%)., Conclusions: The distribution of vaccine serotypes in the analyzed strains was of 50% for the serotypes present in the seven-valent vaccine, 50.2% for the serotypes present in the ten-valent vaccine and 57.2% for those present in the thirteen-valent vaccine.

Objectives: To determine the frequency and clinical features of central nervous system infections caused by enterovirus in children treated at the Hospital Nacional Cayetano Heredia in Lima, Peru., Materials and Methods: A prospective, descriptive study was performed from April 2008 to March 2010. Patients aged 1 month - 14 years with clinical diagnosis of encephalitis or aseptic meningitis were included. We investigated the presence of enterovirus, herpes simplex virus 1 (HSV-1), herpes simplex virus 2 (HSV-2) and varicella-zoster virus (VZV) by polymerase chain reaction (PCR)., Results: 97 patients were included, out of which 69 % had acute encephalitis and 31 % acute meningitis. Enteroviruses were identified in 52,6% of all acute non-bacterial central nervous system infections; corresponding to 83,3 % of meningitis and 38,8 % of encephalitis. There were no cases of infection due to HSV-1, HSV-2 or VZV. Enterovirus infections reached 82,9 % in the warm months (November-January) and 28,6 % in the colder months (May-July)., Conclusions: Enteroviruses are the principal etiologic agents in acute aseptic meningitis and encephalitis in pediatric patients in Lima, Peru. Enteroviruses have a seasonal epidemiological pattern with a clear increase in the number of cases during the summer months. It is useful to have this rapid diagnostic method available as an aid in the management of acute central nervous system infections.

Antibiotic resistance is an emerging problem worldwide present in many bacteria, specially in Escherichia coli, which has high percentages of resistance to ampicilline, thrimethoprim-sulfamethoxazole, tetracycline, chloramphenicol and nalidixic acid, which implies important complications in antibiotic treatment when required. The increasing antibiotic resistance is due to the acquisition of different molecular mechanisms of resistance through point chromosomal mutations and /or horizontal transfer of genetic material between related or different species facilitated by some genetic elements such as integrons. This review discusses the effects of the most common molecular mechanisms of antibiotic resistance in E. coli: enzymatic inactivation, changes in the target site and permeability disturbances. Getting to know the mechanisms of resistance which are involved, as the World Health Organization recommends, will allow us to improve the surveillance of the antibiotic resistance, the control policies and the antibiotic utilization at a national level.

Unlabelled: INTRODUCTION; Diarrheagenic E. coli (DEC) are a major cause of diarrhea in children in developing countries. However, they are not part of routine diagnosis in clinical laboratories., Objectives: To determine the DEC prevalence in Peruvian children and to describe the genetic variability of these strains., Materials and Methods: A total of 8 003 E. coli strains previously isolated from eight different studies of diarrhea in children, mainly from peri-urban areas of Lima, were analyzed. Diagnosis of DEC was done with Multiplex real-time PCR using genes for each of the 6 DEC groups. Conventional PCR was performed for the detection of additional virulence genes., Results: Globally, the mean prevalence in diarrhea samples (n=4,243) was: enteroaggregative E. coli (EAEC) 9.9%, enteropathogenic E. coli (EPEC) 8.5%, enterotoxigenic E. coli (ETEC) 6.9%, diffusely adherent E. coli (DAEC) 4.8%, Shiga toxin-producing E. coli (STEC) 0.8% and enteroinvasive E. coli (EIEC) 0.6%. The relative frequency of each pathogen varies according to the age and the type of study. The main pathotypes in control samples (n=3,760) were EPEC (10.9%) and EAEC (10.4%). An important variability in the virulence genes frequency and molecular resistance mechanisms for each pathotype was found, without differences between diarrhea and control groups., Conclusions: DEC are a major cause of diarrhea in Peruvian children. These pathogens are highly heterogeneous. Additional studies are required to determine the prevalence in rural areas of Peru and in severe diarrhea cases.

Introduction: Diffusely adherent E. coli (DAEC) is the sixth recognized group of diarrheagenic E. coli. However, its association with diarrhea remains controversial. Variability in the adherence patterns of clinical strains is unknown., Objectives: To compare the adherence patterns between strains isolated from children with and without diarrhea., Materials and Methods: A total of 31 DAEC strains were analyzed, 25 from children with diarrhea and 6 from asymptomatic (control) children, isolated from a cohort study of children under one year of age in the southern districts of Lima. DAEC were identified by PCR (daaD gene). The pattern and adherence score in HEp-2 cell culture were evaluated, Actin polimerization was determined by fluorescence actin staining (FAS) and motility was evaluated by conventional microbiology methods., Results: Diffuse adherence pattern was found in 88% of diarrhea samples and in the total of control strains. The number of bacteria adhered per cell was significantly lower in diarrhea samples (p<0.05). However, actin polymerization was greater in diarrhea samples (60% vs. 17%). Motility test was positive in 60% of the diarrhea samples and in all control samples., Conclusions: Our findings suggest a difference between adherence patterns, actin polymerization and motility between DAEC strains corresponding to diarrhea and control groups. The significance of these results must be confirmed with a bigger number of strains and determining the presence of virulence genes in the strains.