Increase of Macrolide-Resistance in Streptococcus pneumoniae Strains After the Introduction of the 13-Valent Pneumococcal Conjugate Vaccine in Lima, Peru.

Streptococcus pneumoniae upper respiratory infections and pneumonia are often treated with macrolides, but recently macrolide resistance is becoming an increasingly important problem. The 13-valent pneumococcal conjugate vaccine (PCV13) was introduced in the National Immunization Program of Peru in 2015. This study aimed to evaluate the temporal evolution of macrolide resistance in S. pneumoniae isolates collected in five cross-sectional studies conducted before and after this vaccine introduction, from 2006 to 2019 in Lima, Peru. A total of 521 and 242 S. pneumoniae isolates recovered from nasopharyngeal swabs from healthy carrier children < 2 years old (2 carriage studies) and samples from normally sterile body areas from pediatric patients with invasive pneumococcal disease (IPD) (3 IPD studies), respectively, were included in this study. Phenotypic macrolide resistance was detected using the Kirby-Bauer method and/or MIC test. We found a significant increase in macrolide resistance over time, from 33.5% to 50.0% in carriage studies, and from 24.8% to 37.5% and 70.8% in IPD studies. Macrolide resistance genes [erm(B) and mef(A/E)] were screened using PCR. In carriage studies, we detected a significant decrease in the frequency of mef(A/E) genes among macrolide-resistant S. pneumoniae strains (from 66.7% to 50.0%) after introduction of PCV13. The most common mechanism of macrolide-resistant among IPD strains was the presence of erm(B) (96.0%, 95.2% and 85.1% in the 3 IPD studies respectively). Macrolide resistance was more common in serotype 19A strains (80% and 90% among carriage and IPD strains, respectively) vs. non-serotype 19A (35.5% and 34.4% among carriage and IPD strains, respectively). In conclusion, S. pneumoniae macrolide resistance rates are very high among Peruvian children. Future studies are needed in order to evaluate macrolide resistance trends among pneumococcal strains, especially now after the COVID-19 pandemic, since azithromycin was vastly used as empiric treatment of COVID-19 in Peru.

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 Humans, Animals, Water and Household Environs in Rural Andean Peru: Exploring Dissemination Pathways through the One Health Lens.

Antimicrobial resistance (AMR) is a global public health threat, especially for low and middle-income countries (LMIC) where the threat has not been fully identified. Our study aims to describe E. coli AMR in rural communities to expand our knowledge on AMR bacterial contamination. Specifically, we aim to identify and describe potential dissemination routes of AMR-carrying bacteria in humans (children's stools), community water sources (reservoirs and household sources), household environments (yard soil) and domestic animals of subsistence farmers in rural Andean areas. Our cross-sectional study was conducted in rural households in the region of Cajamarca, Peru. A total of 266 samples were collected. Thirty-four point six percent of reservoir water and 45% of household water source samples were positive for thermotolerant coliforms. Of the reservoir water samples, 92.8% were positive for E. coli, and 30.8% displayed resistance to at least one antibiotic, with the highest resistance to tetracycline. E. coli was found in 57.1% of the household water sources, 18.6% of these isolates were multidrug-resistant, and displayed the highest resistance to tetracycline (31.3%). Among samples from the children's drinking water source, 32.5% were positive for thermotolerant coliforms, and 57.1% of them were E. coli. One third of E. coli isolates were multidrug-resistant and displayed the highest AMR to tetracycline (41.6%) and ampicillin (25%). Thermotolerant coliforms were found in all the soil samples, 43.3% of the isolates were positive for E. coli, 34.3% of the E. coli isolates displayed AMR to at least one antibiotic, and displayed the highest AMR to tetracycline (25.7%). We determined thermotolerant coliforms in 97.5% of the child feces samples; 45.3% of them were E. coli, 15.9% displayed multidrug resistance, and displayed the highest resistance to ampicillin (34.1%). We identified thermotolerant coliforms in 67.5% of the animal feces samples. Of those, 38.7% were E. coli, and 37.7% were resistant to at least one antibiotic. For all the samples, the prevalence of resistance to at least one antibiotic in the E. coli and Klebsiella spp. isolates was almost 43% and the prevalence of MDR in the same isolates was nearly 9%, yet the latter nearly doubled (15.9%) in children's stools. Our results provide preliminary evidence for critical pathways and the interconnectedness of animal, human and environmental transmission but molecular analysis is needed to track dissemination routes properly.

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 / Frequency of serine protease autotransporters of enterobacteriaceae (SPATE) encoding genes in diffusely adherent escherichia coli (DAEC) isolates from children with and without diarrhea

RESUMEN 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.

ABSTRACT 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.

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 / Frequency of serine protease autotransporters of enterobacteriaceae (SPATE) encoding genes in diffusely adherent escherichia coli (DAEC) isolates from children with and without diarrhea

RESUMEN 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.

ABSTRACT 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.