Pathogenicity Island O-122 in enteropathogenic Escherichia coli strains is associated with diarrhea severity in children from Lima Peru.

EPEC is an attaching and effacing diarrheal pathogen that carries a large pathogenicity island, locus for enterocyte effacement (LEE). Recently, the pathogenicity island PAI O-122 was described among non-LEE effectors and found to be associated with diarrhea among atypical EPEC strains. It is unknown if incomplete PAI O-122 could be associated with diarrhea duration and severity. To identify these virulence determinants we analyzed 379 EPEC strains isolated from Peruvian children. EPEC was diagnosed by PCR(eae+, stx-) and classified as typical(t-EPEC) or atypical(a-EPEC). To characterize PAI O-122 we amplified three modules by PCR: Module 1(pagC), Module 2(senA, nleB and nleE) and Module 3(lifA/efa-1). To characterize the large ORF lifA/efa-1 we amplified the regions known as efa-N, efa-M and efa-C. Clinical information was obtained from the cohort study. A total of 379 EPEC strains were able to analyze PAI O-122 genes, 128 (10.4%) EPEC strains were isolated from 1235 diarrhea episodes and 251(9.2%) from 2734 healthy controls. t-EPEC strains were isolated from 14.8% (19/128) of children with diarrhea and 25/251(10.0%) from healthy controls. The most frequent PAI O-122 genes were nleE(37.7%), senA(34.6%) and nleB(37.5%), with similar prevalence among diarrhea and control samples. However, lifA/efa-1 was more common among diarrhea cases than healthy control cases (30.5% vs. 21.1%, p<0.05). The presence of complete PAI O-122 was associated with diarrhea episodes of higher severity among single pathogen infection (33.3% vs. 1.8%, p<0.05) mainly due to the presence of a complete lifA/efa-1 gene. In summary, the gene lifA/efa-1 is significantly associated with diarrheal episodes of higher severity, suggesting to be an important virulent factor.

Virulence factors and mechanisms of antimicrobial resistance in Shigella strains from periurban areas of Lima (Peru).

The study was aimed to describe the serotype, mechanisms of antimicrobial resistance, and virulence determinants in Shigella spp. isolated from Peruvian children. Eighty three Shigella spp. were serogrouped and serotyped being established the antibiotic susceptibility. The presence of 12 virulence factors (VF) and integrase 1 and 2, along with commonly found antibiotic resistance genes was established by PCR. S. flexneri was the most relevant serogroup (55 isolates, 66%), with serotype 2a most frequently detected (27 of 55, 49%), followed by S. boydii and S. sonnei at 12 isolates each (14%) and S. dysenteriae (four isolates, 5%). Fifty isolates (60%) were multi-drug resistant (MDR) including 100% of S. sonnei and 64% of S. flexneri. Resistance levels were high to trimethoprim-sulfamethoxazole (86%), tetracycline (74%), ampicillin (67%), and chloramphenicol (65%). Six isolates showed decreased azithromycin susceptibility. No isolate was resistant to nalidixic acid, ciprofloxacin, nitrofurantoin, or ceftriaxone. The most frequent resistance genes were sul2 (95%), tet(B) (92%), cat (80%), dfrA1 (47%), blaOXA-1like (40%), with intl1 and intl2 detected in 51 and 52% of the isolates, respectively. Thirty-one different VF profiles were observed, being the ipaH (100%), sen (77%), virA and icsA (75%) genes the most frequently found. Differences in the prevalence of VF were observed between species with S. flexneri isolates, particularly serotype 2a, possessing high numbers of VF. In conclusion, this study highlights the high heterogeneity of Shigella VF and resistance genes, and prevalence of MDR organisms within this geographic region.

Diarrheagenic Escherichia coli phylogroups are associated with antibiotic resistance and duration of diarrheal episode.

Conventionally, in Escherichia coli, phylogenetic groups A and B1 are associated with commensal strains while B2 and D are associated with extraintestinal strains. The aim of this study was to evaluate diarrheagenic (DEC) and commensal E. coli phylogeny and its association with antibiotic resistance and clinical characteristics of the diarrheal episode. Phylogenetic groups and antibiotic resistance of 369 E. coli strains (commensal strains and DEC from children with or without diarrhea) isolated from Peruvian children <1 year of age were determined by a Clermont triplex PCR and Kirby-Bauer method, respectively. The distribution of the 369 E. coli strains among the 4 phylogenetic groups was A (40%), D (31%), B1 (21%), and B2 (8%). DEC-control strains were more associated with group A while DEC-diarrhea strains were more associated with group D (P < 0.05). There was a tendency (P = 0.06) for higher proportion of persistent diarrhea (≥ 14 days) among severe groups (B2 and D) in comparison with nonsevere groups (A and B1). Strains belonging to group D presented significantly higher percentages of multidrug resistance than the rest of the groups (P > 0.01). In summary, DEC-diarrhea strains were more associated with group D than strains from healthy controls.

Class 1 and 2 Integrons in Escherichia coli Strains Isolated from Diarrhea and Bacteremia in Children Less Than 2 Years of Age from Peru.

Class 1 and Class 2 integrons are mobilizable elements able to carry a variety of antibiotic resistance determinants. In the present study, Class 1 and 2 integrons present in 355 pathogenic Escherichia coli (285 diarrheagenic, of these 129 were enteropathogenic, 90 enteroaggregative, 66 enterotoxigenic, and 70 bacteremic) isolated from healthy and ill children under age 5 from periurban areas of Lima, Peru, were characterized. The presence of integrase 1 and 2 was established by polymerase chain reaction (PCR), and variable regions were grouped by PCR-restriction fragment length polymorphism and subsequent sequencing. Antimicrobial resistance was established by disk diffusion. Ninety-seven isolates (27.3%) presented integrase 1, and 16 (4.5%) presented integrase 2 (P < 0.0001); in addition, seven (2.0%) isolates, six diarrheagenic and one bacteremic, presented both integrase genes. The presence of integrase 1 was more frequent among bacteremic isolates (P = 0.0004). Variable regions were amplified in 76/120 (63.3%) isolates with up to 14 gene arrangements. The most prevalent gene cassettes were those encoding dihydrofolate reductases as well as aminoglycoside modifying enzymes. Of note, Class 1 integrons tended to be associated with the presence of extended-spectrum ß-lactamases (ESBLs). A variety of Class 1 and 2 integrons were detected in diarrheagenic and bacteremic E. coli, demonstrating the heterogeneity of variable regions circulating in the area. The association of integrons with ESBLs is worrisome and has an impact on the development of multidrug resistance.

Colonizing and Virulence Factors in Enterotoxigenic Escherichia coli from Peru.

Enterotoxigenic Escherichia coli (ETEC) ranks among the most relevant diarrheagenic pathogens. Efforts to design vaccines to fight ETEC have been focused on colonizing factors (CFs) and atypical virulence factors (AVF). An effective vaccine must account for differences in the regional prevalence of these CFs and AVFs to be truly effective in a given area. In the present study, the presence of 16 CFs and 9 AVFs, as well as the heat-stable (ST) variants (STh or STp), was established by polymerase chain reaction in 205 Peruvian ETEC isolates (120 from diarrhea cases and 85 from healthy controls). Ninety-nine (48.3%) isolates were heat-labile, 63 (30.7%) ST, and 43 (21.0%) presented both toxins. Of ST isolates, 59 (28.8%) possessed STh, 30 (14.6%) STp, five (2.4%) both STh and STp, and 12 (5.8%) were not amplified for any variant tested. The presence of CFs was associated with diarrhea (P < 0.0001). The presence of eatA as well as concomitant presence of CSI, CS3, and CS21 and of C5 and C6 was statistically related to diarrhea cases. The present results suggests that, if effective, a vaccine considering CS6, CS20, and CS21, together with EtpA, would provide protection against 64.4% of the isolates analyzed, whereas the addition of CS12 and EAST1 would lead to 83.9% coverage. Large studies are needed to establish both the ideal candidates to be considered to develop a vaccine effective in the area, and continuous surveillance is needed to detect displacement of circulating isolates that may compromise future vaccines.

First report of CrpP prevalence in a South American country.

The presence of crpP was established in 201 Pseudomonas aeruginosa isolates from 9 Peruvian hospitals. The 76.6% (154/201) of the isolates presented the crpP gene. Overall, 123/201 (61.2%) isolates were non-susceptible to ciprofloxacin. The prevalence of crpP-possessing P. aeruginosa in Peru is higher than in other geographical areas.

High Prevalence of blaCTX-M in Fecal Commensal Escherichia coli from Healthy Children.

BACKGROUND: Antibiotic-resistant Escherichia coli can colonize the intestinal tract of healthy children, causing concern when antibiotic resistance is related to the presence of transferable mechanisms, such as extended-spectrum ß-lactamases (ESBLs). MATERIALS AND METHODS: Fecal samples from 41 healthy children from two villages of rural Peru were cultured on ceftriaxone-disks. ESBL production was confirmed with double disk synergy. In all ESBL-produced isolates, antibiotic susceptibility to 12 antibacterial agents was established by disk diffusion, while clonal relationships were determined by repetitive extragenic palindromic-polymerase chain reaction (REP-PCR). Presence of ST131 was determined using PCR. RESULTS: Ceftriaxone-resistant microorganisms were recovered from 39 samples belonging to 22 out of 41 children (53.7%). Of these, 80 ceftriaxone-resistant and two ceftriaxone-intermediate E. coli from inside ceftriaxone-halos were confirmed as ESBL-producers. All isolates were multidrug-resistant. In 79/80 (98.8%) ceftriaxone-resistant isolates, the presence of blaCTX-M was detected alone (58 isolates, or together with other ß-lactamase (blaTEM, 17 isolates; blaOXA-1-like, 3 isolates; blaTEM + blaOXA-1-like, 1 isolate), while in one isolate no such ESBL was identified. The two ceftriaxone-intermediate isolates recovered from the same sample, carried a blaTEM and blaSHV respectively. Thirty-four different clones were identified, with 4 clones being recovered from different samples from the same child. Twelve clones were disseminated among different children, including 5 clones disseminated between both villages. Two clones, accounting for 3 isolates and both recovered from the same children, belonged to E. coli ST131. CONCLUSION: This study demonstrates high prevalence of ESBL-carriers among healthy children living in a rural area of Peru, stressing the need for continuous surveillance and search for public health control measures.

Whole-Genome Characterisation of ESBL-Producing E. coli Isolated from Drinking Water and Dog Faeces from Rural Andean Households in Peru.

E. coli that produce extended-spectrum ß-lactamases (ESBLs) are major multidrug-resistant bacteria. In Peru, only a few reports have characterised the whole genome of ESBL enterobacteria. We aimed to confirm the identity and antimicrobial resistance (AMR) profile of two ESBL isolates from dog faeces and drinking water of rural Andean households and determine serotype, phylogroup, sequence type (ST)/clonal complex (CC), pathogenicity, virulence genes, ESBL genes, and their plasmids. To confirm the identity and AMR profiles, we used the VITEK®2 system. Whole-genome sequencing (WGS) and bioinformatics analysis were performed subsequently. Both isolates were identified as E. coli, with serotypes -:H46 and O9:H10, phylogroups E and A, and ST/CC 5259/- and 227/10, respectively. The isolates were ESBL-producing, carbapenem-resistant, and not harbouring carbapenemase-encoding genes. Isolate 1143 ST5259 harboured the astA gene, encoding the EAST1 heat-stable toxin. Both genomes carried ESBL genes (blaEC-15, blaCTX-M-8, and blaCTX-M-55). Nine plasmids were detected, namely IncR, IncFIC(FII), IncI, IncFIB(AP001918), Col(pHAD28), IncFII, IncFII(pHN7A8), IncI1, and IncFIB(AP001918). Finding these potentially pathogenic bacteria is worrisome given their sources and highlights the importance of One-Health research efforts in remote Andean communities.

Frequency of serine protease autotransporters of enterobacteriaceae (SPATE) encoding genes in diffusely adherent escherichia coli (DAEC) isolates from children with and without diarrhea. / Frecuencia de genes que codifican proteínas autotransportadoras serin-proteasa de Enterobacteriaceae (SPATE) en cepas de Escherichia coli difusamente adherente (DAEC) provenientes de niños con y sin diarrea.

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.

El objetivo del estudio fue determinar la frecuencia de seis genes que codifican proteínas autotransportadoras serin-proteasa de Enterobacteriaceae (SPATE) en aislamientos de Escherichia coli difusamente adherente (DAEC) provenientes de niños con diarrea (NCD, n=63) y sin diarrea (NSD, n=41) de Lima, Perú. Los NSD se consideraron como grupo control. Para la detección de los genes se estandarizaron 2 PCRs múltiples: triple A (sigA, pet, espP) y triple B (sat, pic, espC). En ambos grupos el gen SPATE más frecuente fue sat (39,7% de NCD y 41,5% de NSD), seguido de espP (20,6% y 9,7% en NCD y NSD respectivamente). Los otros genes se detectaron en proporciones inferiores al 10,0%, en el siguiente orden de frecuencia: pet, sigA, espC y pic, sin diferencias significativas entre los grupos. Se concluye que Sat es la SPATE más frecuente en cepas DAEC, y que estas cepas pueden poseer genes SPATE independientemente de si se aíslan en NCD o NSD.

Antimicrobial Resistance in Rural Settings in Latin America: A Scoping Review with a One Health Lens.

Antimicrobial resistance (AMR) in rural Latin America is not fully understood. The transmission pathways are partially known since research predominantly focuses on the urban hospital setting. The contribution to AMR from environmental factors is usually only mentioned in large-scale animal production. To understand the state of the literature on AMR in rural LA, we carried out a scoping review using the One Health (OH) perspective. OH recognises the concomitant contributions and interconnectedness of humans, animal, and the environment, thus, we used the OH perspective to select those articles adopting a holistic view of the problem. We searched original articles in English, Spanish, and Portuguese in four peer-reviewed databases and included 21 publications in the analysis. We charted data on bibliometrics, design, data collection sources, and instruments. We identified the human, animal, and environmental contributions to AMR in rural locations, and information gaps on AMR transmission routes and AMR drivers. Intensive and non-intensive animal production systems and agricultural practices were the most frequently found human contributions to AMR. Poultry, swine, cattle, and fish were the most frequent livestock mentioned as sources of AMR bacteria. Animal carriage and/or transfer of AMR determinants or bacteria was recognised as the primary contribution of livestock to the problem, while water, soil, and farming were predominant environmental contributions. We found that only 1 article out of 21 considered the OH approach as a framework for their sampling scheme, whereas 5 out 21 discussed all the three OH components. There were hardly any descriptions of humans or human waste as reservoirs for AMR in rural locations, and rural health centres or hospitals and wildlife were not represented. No studies identified mining as an anthropogenic activity driving AMR. More OH-oriented studies, with emphasis on molecular approaches-for identification and comparison of AMR genes-are sorely needed to understand better the existence of a network of interconnected transmission routes in rural Latin America and provide efficient strategies to prevent further AMR emergence.

Dissemination of a multidrug resistant CTX-M-65 producer Salmonella enterica serovar Infantis clone between marketed chicken meat and children.

The objective of the present study was to characterize Salmonella enterica serovar Infantis isolated from chicken meat determining their clonal relationships with S. Infantis isolated from children with diarrhea. Fifteen meat-recovered S. Infantis were analyzed. Susceptibility levels to 14 antibacterial agents, the presence of ESBL and that of inducible plasmid-mediated AmpC (i-pAmpC) were determined by phenotypical methods. The presence of ESBL and pAmpC was confirmed by PCR, and detected ESBL-encoding genes were sequenced and their transferability tested by conjugation. The presence of gyrA mutations as well as Class 1 integrons was determined by PCR. Clonal relationships were established by REP-PCR and RAPD. In addition, 25 clinical isolates of S. Infantis were included in clonality studies. All meat-recovered S. Infantis were MDR, showing resistance to ampicillin, nitrofurans and quinolones, while none was resistant to azithromycin, ceftazidime or imipenem. ESBL (blaCTX-M-65) and i-pAmpC (blaDHA) were detected in 2 and 5 isolates respectively (in one case concomitantly), with blaCTX-M-65 being transferable through conjugation. In addition, 1 isolate presented a blaSHV gene. All isolates presented D87Y at GyrA, nalidixic acid active efflux pump and a Class 1 integron of ~1000 bp (aadA1). Clonal analysis showed that all isolates were related. Further they were identical to MDR blaCTX-M-65-producing S. Infantis isolates causing children diarrhea in Lima. The dissemination of MDR blaCTX-M-65-producing S. Infantis between marketed meat and children highlights a public health problem which needs be controlled at livestock level.

[Comparison of enteropathogenic Escherichia coli (EPEC) diagnosis by serology and by polymerase chain reaction (PCR)]. / Comparación entre el diagnóstico serológico y el diagnóstico por reación en cadena de la polimerasa (PCR) para Escherichia coli enteropatogénica (EPEC).

INTRODUCTION: The identification of EPEC in clinical laboratories is based on the determination of the serotypes by agglutination with O and H antiserum. Currently the proper diagnosis of EPEC should be done by the identification of the intimin gen (eaeA) by PCR. OBJECTIVES: To compare the diagnosis of EPEC by serotyping and by PCR. MATERIALS AND METHODS: We collected EPEC strains, identify by their O antigen, from 4 clinical laboratories in Lima from diarrheal samples in children less than 5 years of age. In those strains we have searched for virulence genes by a real time multiplex PCR for the diarrheagenic E. coli. RESULTS: We collected 113 strains; 82% from children less than 2 years of age. Only 15 strains (13.3%) had the intimin gene and therefore a confirmatory diagnosis of EPEC. In addition we found 3 enterotoxigenic (ETEC), 3 shiga toxin-producing (STEC), 1 enteroagreggative (EAEC) and 1 enteroinvasive (EIEC) strains. CONCLUSIONS: PCR should be use for the proper identification of EPEC. However, molecular methods are still not easily available in clinical laboratories worldwide.

[Antimicrobial resistance of commensal Escherichia coli strains in children of two rural communities in Peru]. / Resistencia antimicrobiana de cepas comensales de Escherichia coli en niños de dos comunidades rurales peruanas.

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.

La resistencia a antibióticos es un problema mundial importante. El objetivo de este estudio fue determinar la resistencia a los antibióticos en cepas comensales aisladas de niños sanos de comunidades rurales de Moyobamba y Urubamba en Perú. Este estudio de cohorte identificó 179 cepas de Escherichia coli comensales provenientes de 93 niños, seguidos durante seis meses. Trece antibióticos fueron analizados mediante disco difusión. Los mayores índices de resistencia fueron para cotrimoxazol (49,1%), ampicilina (48,0%) y ácido nalidíxico (31,8%). Se encontró un aumento en la resistencia del 11,6% para el ácido nalidíxico y del 6,4% para el cotrimoxazol en este periodo. El 34,0% de los aislados fueron multidrogoresistentes. Este estudio respalda hallazgos previos de resistencia a múltiples fármacos en cepas comensales en comunidades rurales, y destaca el aumento en las tasas de resistencia en el tiempo. Recomendamos estudios en poblaciones más grandes, con un seguimiento más prolongado.

Parental Antibiotic Use in Urban and Peri-Urban Health Care Centers in Lima: A Cross-Sectional Study of Knowledge, Attitudes, and Practices.

In pediatric patients, the antibiotic use is affected by parental beliefs and practices; especially in countries where it is possible to acquire them without prescription. This study aims to describe the knowledge, attitudes, and practices on antibiotic use among parents of children from urban and peri-urban health care centers in Lima. A cross-sectional study was performed at 1 urban and 2 peri-urban health care centers selected in Lima, Perú. Parents of children below the age of 3 years answered a knowledge-attitudes-practices-validated questionnaire about antibiotic use and were categorized as high, moderate, and low knowledge regarding antibiotics. We analyzed potential determinants for low knowledge and having medicated their children with unprescribed antibiotics using bivariate and multivariate analyses. A total of 224 parents were enrolled, and 8% were categorized as low knowledge. Half of the parents could not recognize that antibiotics cannot cure viral infections, 59.4% disagreed with "antibiotics speed up recovery from a cold," and 53.2% stored antibiotics at home. Remarkably 23.5% of parents reported having medicated their children with antibiotics without prescription, which was associated with belonging to the peri-urban health care center, use of antibiotics by their children in the last 12 months, and having purchased antibiotics without physicians' prescription. An alarming overuse of antibiotics without prescription was described among children below the age of 3 years. Educational interventions, addressing parental attitudes and practices, and health policies should be developed to limit inappropriate antibiotic use especially in peri-urban communities.

Antibiotic-Resistant Escherichia coli in Drinking Water Samples from Rural Andean Households in Cajamarca, Peru.

Antibiotic resistance in pathogenic bacteria is a serious public health issue. The growing threat is a cause for concern and action to prevent the emergence of new resistant strains and the spread of existing ones to humans via the environment. This study aimed at identifying fecal pathogens in drinking water obtained from rural Andean households from Cajamarca, Peru, and measuring the antibiotic resistance profile of Escherichia coli. The study was embedded within a community-randomized controlled trial among 102 communities in the northern highlands of the Cajamarca region, Peru. Of 314 samples, 55.4% (95% CI [49.7, 61.0], n = 174) were identified as thermotolerant coliforms. Among the samples positive for thermotolerant coliform, E. coli was isolated in 37.3% (n = 117), Klebsiella spp. in 8.0% (n = 25), Enterobacter spp. in 5.1% (n = 16), and Citrobacter spp. in 2.5% (n = 8). Of the 117 E. coli samples, 48.7% (95% CI [39.4, 58.1], n = 57) showed resistance to any antibiotic. The E. coli antibiotic resistance profile showed highest resistance against tetracycline (37.6%), ampicillin (34.2%), sulfamethoxazole-trimethoprim (21.4%), and nalidixic acid (13%). Some 19.7% (95% CI [12.9, 28.0], n = 23) of the E. coli isolates displayed multidrug resistance, defined as resistance to at least three classes of antibiotics. The CTX-M-3 gene, which encodes extended-spectrum resistance to beta-lactamase antibiotics, was found in one isolate. The high prevalence of fecal contamination in drinking water highlights the importance of household water treatment methods. Likewise, the high levels of antibiotic resistance found indicate a need for further research to identify the origins of potential environmental contamination, misuse, or inadequate disposal of antibiotics.

Stunting Is Preceded by Intestinal Mucosal Damage and Microbiome Changes and Is Associated with Systemic Inflammation in a Cohort of Peruvian Infants.

Stunting, defined as height-for-age Z score equal to or lower than -2, is associated with increased childhood mortality, cognitive impairment, and chronic diseases. The aim of the study was to investigate the relationship between linear growth, intestinal damage, and systemic inflammation in infants at risk of stunting. We followed up 78 infants aged 5-12 months living in rural areas of Peru for 6 months. Blood samples for biomarkers of intestinal damage (intestinal fatty-acid-binding protein [I-FABP] and zonulin) and systemic inflammation (interleukin-1ß, interleukin-6, tumor necrosis factor α [TNF-α], soluble CD14, and lipopolysaccharide-binding protein [LBP]) and fecal samples for microbiome analysis were collected at baseline and closure of the study. The children's growth and health status were monitored through biweekly home visits by trained staff. Twenty-one percent of the children became stunted: compared with non-stunted children, they had worse nutritional parameters and higher levels of serum I-FABP at baseline. The likelihood of becoming stunted was strongly associated with an increase in sCD14 over time; LBP and TNF-α showed a trend toward increase in stunted children but not in controls. The fecal microbiota composition of stunted children had an increased beta diversity compared with that of healthy controls throughout the study. The relative abundance of Ruminococcus 1 and 2, Clostridium sensu stricto, and Collinsella increased in children becoming stunted but not in controls, whereas Providencia abundance decreased. In conclusion, stunting in our population was preceded by an increase in markers of enterocyte turnover and differences in the fecal microbiota and was associated with increasing levels of systemic inflammation markers.

[Presence of multidrug resistant Enterobacteriaceae and Escherichia coli in meat purchased in traditional markets of Lima]. / Presencia de Enterobacteriaceae y Escherichia coli multirresistente a antimicrobianos en carne adquirida en mercados tradicionales en Lima.

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.

OBJETIVOS.: El objetivo del presente estudio fue describir la presencia de Enterobacteriaceae en muestras de carne recolectadas en mercados tradicionales de Lima y establecer los niveles de resistencia a antimicrobianos y la presencia de betalactamasas de espectro extendido (BLEE) y AmpC en Escherichia coli. MATERIALES Y MÉTODOS.: Se recolectaron un total de 138 muestras de carne, 64 (46,4 %) de pollo, 44 (31,9 %) de carne de res y 30 (21,7%) de carne de cerdo. Las bacterias aisladas pertenecieron a 17 géneros diferentes, y específicamente 14 fueron clasificados como Enterobacteriaceae. Se analizó la sensibilidad frente a diez agentes antimicrobianos mediante el método de difusión de disco Kirby-Bauer, se determinó la presencia de BLEE y AmpC mediante las pruebas de doble disco y de inducción de imipenem-ceftazidima, respectivamente. RESULTADOS.: Los niveles de resistencia a los antimicrobianos fueron altos frente a trimetoprima-sulfametoxazol, ampicilina, tetraciclina, ácido nalidíxico, ciprofloxacino y cloranfenicol. Existen diferencias significativas en los niveles de resistencia a los antibióticos según el tipo de carne (pollo, carne de res y cerdo) (p <0,05). Los niveles de resistencia a múltiples antimicrobianos (MDR) fueron particularmente altos en pollo y cerdo (98,2 % y 86,4 %, respectivamente). Además, la presencia de BLEE en Escherichia coli aisladas de carne de pollo fue del 59,4 %. CONCLUSIONES.: Los niveles de resistencia a los antimicrobianos fueron altos frente a los antibióticos usados frecuentemente en humanos, se destaca el pollo y la res como potenciales reservorios de Escherichia coli productoras de BLEE y pAmpC, respectivamente.

Virulence and Antimicrobial Resistance in Campylobacter spp. from a Peruvian Pediatric Cohort.

The presence of virulence factors (VFs) and mechanisms of quinolones and macrolide resistance was analyzed in Campylobacter spp. from a pediatric cohort study in Lima. In 149 isolates (39 Campylobacter jejuni and 24 Campylobacter coli from diarrheic cases; 57 C. jejuni and 29 C. coli from controls), the presence of the cdtABC and cadF genes and iam marker was established. Nalidixic acid, ciprofloxacin, erythromycin, and azithromycin susceptibilities were established in 115 isolates and tetracycline-susceptibility was established in 100 isolates. The presence of mutations in the gyrA, parC, and 23S rRNA genes was determined. The cadF gene and all genes from the cdtABC operon were significantly more frequent among C. jejuni (P < 0.0001); the iam marker was more frequent in C. coli (P < 0.0001). No differences were observed in VFs between cases and controls. Almost all isolates were tetracycline-resistant; nalidixic acid and ciprofloxacin resistance reached levels of 90.4% and 88.7%, respectively. Resistance to macrolides was 13% (C. jejuni 4.3%; C. coli 26.1%). Resistance to ciprofloxacin was related to GyrA Thr86 substitutions, while 13 of 15 macrolide-resistant isolates possessed a 23S rRNA mutation (A2075G). Differences in the presence of VFs and alarming levels of resistance to tested antimicrobial agents were observed among C. jejuni and C. coli.

Molecular and Phenotypic Characterization of Diarrheagenic Escherichia coli Strains Isolated from Bacteremic Children.

Escherichia coli is an important cause of Gram-negative bacteremia. The aim of this study was to characterize at the molecular and phenotypic levels E. coli strains belonging to different diarrheagenic pathotypes [diarrheagenic E. coli (DEC)] isolated from bacteremia in children younger than 5 years of age. Seventy bacteremia E. coli strains were collected in a prospective study in 12 hospitals in Lima, Peru. The presence of virulence genes associated with DEC [enterotoxigenic (lt and st), enteropathogenic (eaeA), shiga toxin-producing (stx1and stx2), enteroinvasive (ipaH), enteroaggregative (aggR), and diffusely adherent (daaD)] was determined by multiplex real-time polymerase chain reaction (PCR). Those positive E. coli strains were further analyzed for 18 additional virulence factors encoding genes and others phenotypic features. Virulence genes associated with DEC were identified in seven bacteremic children (10%), including: one aggR-positive [enteroaggregative E. coli (EAEC)], one eaeA-positive [enteropathogenic E. coli (EPEC)], one st-positive [enterotoxigenic E. coli (ETEC)], one daaD-positive [diffusely adherent E. coli (DAEC)], and three strain positive for aggR and daaD (EAEC/DAEC) at the same time. All strains, except EPEC, had the Ag43 adhesin, and all, except ETEC had the siderophore gene fyuA. The phylogenetic profile of these strains was variable, two (B2), two (D), two (A), and one (B1) strain. These isolates were susceptible to all tested antibacterial agents except to ampicillin and gentamicin. The three EAEC/DAEC strains showed biofilm formation and aggregative adhesion and had the same repetitive extragenic palindromic-PCR patterns. These findings suggest that some DEC strains, especially agg-R and daa-D positive, might cause bacteremia in children.

[Multiresistance and factors associated with the presence of extended-spectrum beta-lactamases in Escherichia coli strains isolated from urine culture]. / Multirresistencia y factores asociados a la presencia de betalactamasas de espectro extendido en cepas de Escherichia coli provenientes de urocultivos.

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.

El tratamiento empírico para la infección urinaria se ve complicado frente a la presencia de multirresistencia y de betalactamasas de espectro extendido (BLEE). El objetivo del estudio fue describir los patrones de resistencia antibiótica de cepas de Escherichia coli aisladas en urocultivos y los factores clínico-epidemiológicos asociados a la presencia de BLEE en un grupo pediátrico y adulto. Se recolectaron durante 14 meses, 353 cepas provenientes de Emergencia y Hospitalización del Hospital Cayetano Heredia, 45,9% fueron cepas multirresistentes. La incidencia de BLEE en población pediátrica fue 16,3% vs. 31,1% en la adulta, el 63,6% provenía de pacientes ambulatorios. La presencia de BLEE se asoció con encontrarse hospitalizado en pediatría, así cómo al uso de pañal y vejiga neurogénica en adultos. Estos factores deben considerarse al momento de elegir un tratamiento antibiótico. Asimismo, es necesario implementar programas de reporte epidemiológico y modelos de prevención de factores de riesgo.