The Crisis of Macrolide Resistance in Pneumococci in Latin America.

Macrolide antibiotics are recommended for the treatment of pneumococcal pneumonia and invasive pneumococcal disease (IPD). Prior to 2000, ∼10% of Streptococcus pneumoniae strains isolated from IPD cases in Latin American countries were resistant to macrolides. The mechanism of resistance to macrolides was associated mainly with the efflux pump known as the macrolide efflux genetic assembly, since most pneumococcal strains carried the mef(A/E) gene, whereas <6% strains carried both the methylase gene ermB and mef(A/E). In the first decade of this century, a significant increase in the prevalence of macrolide resistance was observed in pneumococcal strains in both Mexico and Peru. Approximately 30% of S. pneumoniae strains in these countries were already resistant to erythromycin, while the prevalence in Colombia, Argentina, and Brazil remained below 10%. During the last decade, we have been experiencing a worrisome increase in pneumococcal strains carrying resistance to macrolides, with a prevalence of up to 80% for resistance to erythromycin. The mechanism for disseminating macrolide resistance has evolved. Currently, more than 55% of invasive S. pneumoniae macrolide-resistant strains carry both the ermB and the mef(A/E)/mel genes. Lessons learned from the current macrolide resistance crisis in Latin America can inform interventions in other regions.

Association between nasopharyngeal colonization with multiple pneumococcal serotypes and total pneumococcal colonization density in young Peruvian children.

OBJECTIVES: We examined the association of nasopharyngeal (NP) pneumococcal co-colonization (>1 pneumococcal serotype) and pneumococcal density in young Peruvian children enrolled in a prospective cohort study. METHODS: NP swabs collected monthly from children aged <3 years during both asymptomatic and acute respiratory illness (ARI) periods underwent culture-enriched microarray for pneumococcal detection and serotyping and lytA polymerase chain reaction for density assessment. We examined the serotypes commonly associated with co-colonization and the distribution of densities by co-colonization, age, current ARI, and other covariates. The association of co-colonization and pneumococcal density was assessed using a multivariable mixed-effects linear regression model, accounting for repeated measures and relevant covariates. RESULTS: A total of 27 children contributed 575 monthly NP samples. Pneumococcus was detected in 302 of 575 (53%) samples, and co-colonization was detected in 61 of these 302 (20%). The total densities were higher during ARI than non-ARI periods and lowest among the youngest children, increasing with age. In the multivariable analysis, there was no significant association between pneumococcal density and co-colonization (coefficient estimate 0.22, 95% confidence interval 0.11-0.55; reference: single-serotype detections). Serotypes 23B and 19F were detected significantly more frequently as single isolates. CONCLUSION: Pneumococcal co-colonization was common and not associated with increased pneumococcal density. Differential propensity for co-colonization was observed among individual serotypes.

Pneumococcal Carriage Among Indigenous Kichwa Children From the Ecuadorian Andes After the 10-Valent Pneumococcal Vaccine Introduction.

BACKGROUND: We assessed nasopharyngeal pneumococcal carriage in Andean Kichwa children, the largest Amerindian indigenous population in the Ecuadorian Andes. All children in our study had been vaccinated with the 10-valent pneumococcal vaccine (PCV10). METHODS: Nasopharyngeal swabs from 63 families, 100 children <10 years old including 38 children under 5 years and 63 adult caregivers, from 5 different communities, were cultivated for Streptococcus pneumoniae and isolates were serotyped and antibiotic susceptibility testing was performed. RESULTS: Respectively, 67% of the 38 children under 5 years old, 49% of the 62 children between 6 and 10 years old and 16% of the 100 adults were colonized with S. pneumoniae. Of these, 30.9% carried a vaccine serotype, 5.4% a serotype shared by the PCV10/13-valent pneumococcal vaccine (PCV13) vaccine and 25.5% a PCV13 serotype or PCV13 vaccine-related serotype, with 19A (10.9%) and 6C (10.9%) as the most prominent. Drug susceptibility testing revealed that 46% of the S. pneumoniae strains were susceptible to 6 tested antibiotics. However, 20.3% of the strains were multidrug-resistant or extensively drug-resistant strains, including 82% of the vaccine (-related) serotype 19A and 6C strains. CONCLUSIONS: Kichwa children, vaccinated with PCV10, were highly colonized with pneumococci and should be considered a high-risk group for pneumococcal disease. Twenty-five percent of the colonizing S. pneumoniae strains were PCV13-only vaccine-targeted serotypes, and in addition to that, most were multidrug-resistant or extensively drug-resistant strains. The vaccine benefits for this population possibly will significantly increase with the introduction of PCV13.

Non-diarrheagenic and diarrheagenic E. coli carrying supplementary virulence genes (SVG) are associated with diarrhea in children from Mexico.

Escherichia coli strains, including diarrheagenic E. coli (DEC), are among the most important causes of childhood diarrhea in developing countries. Since these strains also colonize healthy children, additional factors leading to diarrhea remains to be discovered. We therefore conducted a comprehensive study to investigate if supplementary virulence genes (SVG) carried by DEC strains and non-DEC strains, contribute to diarrhea in Mexican children. E. coli strains were isolated from n = 317 children between 6 and 12 years, n = 114 with diarrhea and n = 203 asymptomatic children from Northwestern Mexico, PCR was used to identify SVG, then virulence score and cytotoxic assay in HT-29 cells were performed to evaluate virulence of E. coli strains. DEC prevalence was 18.6% and its presence was significantly associated with diarrhea cases. aEPEC, tEAEC, ETEC, DAEC, aEAEC, tEPEC, and EIEC pathotypes were identified. aEPEC strains were significantly associated with asymptomatic children, whereas ETEC was only identified in children with diarrhea. E. coli strains carrying colonization-related SVG and/or proteolysis-related SVG were significantly associated with diarrhea. DEC strains were associated to diarrhea if strains carried SVG ehaC, kps, nleB, and/or espC. Virulence score was significantly higher in E. coli from diarrhea cases than asymptomatic. In addition, DEC strains carrying SVG+ were more virulent, followed by non-DEC SVG+ strains, and correlated with the cytotoxicity assay. Nearly 50% of DEC strains were MDR, and ~10% were XDR. In conclusion the findings of this work provide evidence that the presence of E. coli strains (regardless if strains are DEC or non-DEC) with SVG were associated with diarrhea in Mexican children.

Pseudomonas aeruginosa Isolates From a Cohort of Mexican Children With Cystic Fibrosis Show Adaptation to a Chronic Phenotype.

BACKGROUND: Long-term persistence of Pseudomonas aeruginosa in the lung of individuals with cystic fibrosis (CF) is associated with progressive selection of diverse genotypes and phenotypes. This bacterial adaptation leads to chronic infection and increased morbidity and mortality. The aim of this study was to establish the prevalence, clonal relatedness, antimicrobial susceptibility and virulence-associated phenotypes of P. aeruginosa isolates in a cohort of 50 Mexican children with CF-associated chronic lung infection. METHODS: Clonal relatedness of P. aeruginosa isolates was verified by pulsed-field gel electrophoresis. The antimicrobial susceptibility was determined by an automated system that performs bacterial identificación and antibiotic susceptibility testing (VITEK 2) and/or broth microdilution method. Biofilm formation was quantified with the crystal violet method; swarming motility was measured on soft agar, and susceptibility to normal human serum determined by reduction of colony formed units (CFUs). RESULTS: High prevalence of P. aeruginosa colonization among Mexican children with CF was confirmed; 20% (10/49) of clones identified showed a multidrug-resistant phenotype and 8.2% (4/49) an extensive drug resistance phenotype; 26.5% (13/49) of the isolates were resistant to colistin, 42.9% (21/49) presented a phenotype of adaptation associated with chronic infection and 79.6% (39/49) showed increased ability to survive in normal human serum. CONCLUSIONS: This cohort of children with CF reveals that colonizing P. aeruginosa strains predominantly display resistance to several first-line antibiotics, although most isolates were susceptible to meropenem and tobramycin; 42.9% of isolates showed a phenotype consistent with adaptation to chronic lung infection.

Captive Green Iguana Carries Diarrheagenic Escherichia coli Pathotypes.

The green iguana appears to be a carrier for bacteria causing gastrointestinal infections in humans. The presence of diarrheagenic E. coli (DEC) pathotypes, however, has not been studied in this reptile. The aim of the current work was to investigate the prevalence of DEC in the intestines of 240 captive green iguanas, their phylogenetic groups, and the antibiotic susceptibility profile. E. coli strains were isolated from 41.7% (N = 100/240) of the intestinal content of green iguanas. DEC strains was identified in 25.9% of the screened population and were detected in the majority (62%, p = 0.009) of those reptiles carrying E. coli strains. Among DEC strains, STEC strains carrying the stx1 gene were the most prevalent pathotype isolated (38.7%), followed by EAEC and ETEC (27.4% each). Genetic markers of DEC strains belonging to the EHEC pathotype were not detected. More than a half of DEC strains were classified into the Clade I-II phylogroup (64.5%), followed by the phylogroup A (14.5%). The antibiotic susceptibility method demonstrated that a high proportion of DEC strains were resistance, or non-susceptible, to carbenicillin, amikacin, and ampicillin. We conclude that the green iguana kept in captivity is a carrier of DEC strains bearing resistance to first-line antibiotics, including penicillins. Given the increase presence of the green iguana in Latin American households, these reptiles represent a potential source of transmission to susceptible humans and therefore a potential source of gastrointestinal disease.

Lactoferrin Disaggregates Pneumococcal Biofilms and Inhibits Acquisition of Resistance Through Its DNase Activity.

Streptococcus pneumoniae colonizes the upper airways of children and the elderly. Colonization progresses to persistent carriage when S. pneumoniae forms biofilms, a feature required for the development of pneumococcal disease. Nasopharyngeal biofilms are structured with a matrix that includes extracellular DNA (eDNA), which is sourced from the same pneumococci and other bacteria. This eDNA also allows pneumococci to acquire new traits, including antibiotic resistance genes. In this study, we investigated the efficacy of lactoferrin (LF), at physiological concentrations found in secretions with bactericidal activity [i.e., colostrum (100 µM), tears (25 µM)], in eradicating pneumococcal biofilms from human respiratory cells. The efficacy of synthetic LF-derived peptides was also assessed. We first demonstrated that LF inhibited colonization of S. pneumoniae on human respiratory cells without affecting the viability of planktonic bacteria. LF-derived peptides were, however, bactericidal for planktonic pneumococci but they did not affect viability of pre-formed biofilms. In contrast, LF (40 and 80 µM) eradicated pneumococcal biofilms that had been pre-formed on abiotic surfaces (i.e., polystyrene) and on human pharyngeal cells, as investigated by viable counts and confocal microscopy. LF also eradicated biofilms formed by S. pneumoniae strains with resistance to multiple antibiotics. We investigated whether treatment with LF would affect the biofilm structure by analyzing eDNA. Surprisingly, in pneumococcal biofilms treated with LF, the eDNA was absent in comparison to the untreated control (∼10 µg/ml) or those treated with LF-derived peptides. EMSA assays showed that LF binds S. pneumoniae DNA and a time-course study of DNA decay demonstrated that the DNA is degraded when bound by LF. This LF-associated DNase activity inhibited acquisition of antibiotic resistance genes in both in vitro transformation assays and in a life-like bioreactor system. In conclusion, we demonstrated that LF eradicates pneumococcal-colonizing biofilms at a concentration safe for humans and identified a LF-associated DNAse activity that inhibited the acquisition of resistance.

Detection of antimicrobial-resistance diarrheagenic Escherichia coli strains in surface water used to irrigate food products in the northwest of Mexico.

Water contamination by pathogenic bacteria is a global public health problem. Contamination of surface water utilized to irrigate food products, or for human consumption, causes outbreaks of foodborne and waterborne disease. Of these, those caused by diarrheagenic Escherichia coli (DEC) strains present substantial morbidity and mortality. The aim of this study was, therefore, to investigate the microbiological quality of surface water and the presence of DEC strains in different water bodies. A total of 472 water samples were collected from irrigation canal, dam, river, and dike water bodies from January through December 2015 in Sinaloa, a State located in Northwestern Mexico. Our studies demonstrated that 47.0% (222/472) of samples contained thermotolerant coliforms above permissive levels whereas E. coli strains were isolated from 43.6% (206/472). Among these E. coli isolates, DEC strains were identified in 14% (29/206) of samples including in irrigation canal (26/29) and river water (3/29) collected from the northern (83%) and central area (17%). Isolated DEC strains were classified as enteroaggregative E. coli (EAEC) 34.4% (10/29), enteropathogenic E. coli (EPEC) 31.0% (9/29), diffuse adherent E. coli (DAEC) 27.5% (8/29), and enterotoxigenic E. coli (ETEC) 6.8% (2/29). Moreover, 90% of isolated DEC strains exhibited resistance to at least one commonly prescribed antibiotic in Mexico whereas 17% were multi-drug resistant. In conclusion, the presence of DEC strains in surface water represents a potential source for human infection, and thus routine monitoring of DEC in surface water and other indirect affected areas should be considered at northwestern Mexico.

Children from a rural region in The Chiapas Highlands, Mexico, show an increased risk of stunting and intestinal parasitoses when compared with urban children.

Background: The state of Chiapas has held the first place of extreme poverty in Mexico. The majority of Chiapas' municipalities are inhabited by marginalized, indigenous populations, who usually present diarrhea of unknown etiology. We evaluated the nutritional status, intestinal parasites, and common bacterial pathogens, including DEC (diarrheagenic Escherichia coli) strains, in 178 children under five years of age with a high (rural) and a moderate (urban) degree of marginalization. Methods: Z-scores for anthropometric indexes from the children were obtained, whereas intestinal parasites were investigated by using a direct coproparasitoscopic analysis and a concentration method. DEC strains were detected by polymerase chain reaction (PCR). Results: The stunting prevalence in children from the rural and urban regions was 79.8 and 7.5%, respectively. Only children from rural municipalities were parasitized (72.6%), being Ascaris lumbricoides and Entamoeba histolytica/Entamoeba dispar the most prevalent parasites (57.1 and 38.1%, respectively). More than half of the children presented moderated ascariasis. Besides Giardia intestinalis, these parasites were associated with stunting. The prevalence of DEC strains was similar in both regions. Conclusions: Only children from the Chiapas Highlands (rural zone) exhibited high prevalences of stunting and intestinal parasites. A reevaluation of social development programs should be in place to address stunting and intestinal parasitoses, mainly in rural regions of Chiapas, to avoid adverse functional consequences on these children.

Introducción: El estado de Chiapas ha ostentado el primer lugar de pobreza extrema en México. La mayor parte de la población de los municipios de Chiapas es indígena, vive en condiciones de marginación y padece de diarrea de etiología desconocida. Este trabajo evaluó el estado nutricional, la presencia de parásitos intestinales y patógenos bacterianos comunes, además de cepas DEC (Escherichia coli diarreogénica) en 178 niños menores de cinco años, provenientes de una localidad con alto grado de marginación (rural) y de una con moderada marginación (urbana). Métodos: Se obtuvieron los puntajes Z de los índices antropométricos de los niños. Los parásitos intestinales se investigaron con el método coproparasitoscópico directo y un método de concentración. Las cepas DEC se detectaron mediante reacción en cadena de la polimerasa. Resultados: La prevalencia de desmedro en niños de la zona rural y urbana fue de 79.8 y 7.5%, respectivamente. Únicamente los niños de la zona rural estuvieron parasitados (72.6%), y los más prevalentes fueron Ascaris lumbricoides y Entamoeba histolytica/Entamoeba dispar (57.1 y 38.1%, respectivamente). Más de la mitad de los infantes exhibieron ascariasis moderada. Estos parásitos, además de Giardia intestinalis, se asociaron con el desmedro. En ambas regiones, la prevalencia de DEC fue similar. Conclusiones: Solo los niños de los Altos de Chiapas (zona rural) exhibieron alta prevalencia de desmedro y parásitos intestinales. Para evitar las consecuencias adversas entre los infantes, es necesario reevaluar los programas de desarrollo social para combatir el desmedro y la parasitosis intestinal, principalmente en las regiones rurales de Chiapas.

Stunting and intestinal parasites in school children from high marginalized localities at the Mexican southeast.

INTRODUCTION: Children under five years of age from developing countries are in risk of not achieving an adequate human development due to stunting and extreme poverty. They were also affected by intestinal helminths. Inhabitants of the state of Chiapas, the poorest population in Mexico, register the highest prevalence of child malnutrition as well as intestinal parasitic infections. With the purpose of fight against poverty and hunger, the Mexican government launched a social program called "Prospera". The aim of this work was to determine the prevalence of stunting and intestinal parasites in school children beneficiaries of that social program, from two marginalized municipalities of Chiapas, Mexico. METHODOLOGY: A total of 106 school-age children were recruited for nutritional assessment as well parasitic load measures. RESULTS: Most children exhibited stunting (88.7%). In these children the prevalence of intestinal parasites was 32.1%, being A. lumbricoides the species with the highest prevalence (25.5%) with moderate parasitic load (15.1%). Positive associations were observed between the presence of intestinal parasites and the municipality where children lived, the type of footwear, or the educational level of the mother. CONCLUSIONS: Extreme poverty conditions in these localities of Mexico are far from reaching the sustainable development goals.

Nosocomial, Multidrug-Resistant Klebsiella pneumoniae Strains Isolated from Mexico City Produce Robust Biofilms on Abiotic Surfaces but Not on Human Lung Cells.

BACKGROUND: Klebsiella pneumoniae (Kpn) strains are a leading cause of hospital-acquired infections, including ventilator-associated pneumonia. Resistance to antibiotics, biofilm formation, and the production of certain fimbriae play an important role in the pathogenesis. AIM: We investigated the genetic relatedness, antibiotic resistance, virulence potential, and ability to form biofilms of Kpn strains isolated from hospital-acquired infections (n = 76). Strains were isolated at three major hospitals serving the largest metropolitan urban area in Mexico City, Mexico. RESULTS: Enterobacterial repetitive intergenic consensus (ERIC)-PCR demonstrated that clonal groups predominate in each hospital. Selected strains chosen from clonal groups (n = 47) were multidrug resistant (MDR, 83%), although the majority (∼70%) were susceptible to carbapenems. All strains produced robust biofilms on abiotic surfaces, and ∼90% harbored adhesin genes fimH, mrkA, and ecpA. The ultrastructure of biofilms was further studied by high-resolution confocal microscopy. The average height of Kpn biofilms on abiotic surfaces was ∼40 µm. We then assessed formation of biofilms on human lung cells, as a surrogate of lung infection. While Kpn strains formed robust biofilms on abiotic surfaces, studies on lung cells revealed attachment to human cells but scarce formation of biofilms. Gene expression studies revealed a differential temporal expression of an adhesin (ecpA) and a capsule (galF) gene when biofilms were formed on different substrates. CONCLUSIONS: Kpn strains isolated from nosocomial infections in Mexico City are MDR, although the majority are still susceptible to carbapenems and form more robust biofilms on polystyrene in comparison to those formed on human cells.

Surveillance of Diarrheagenic Escherichia coli Strains Isolated from Diarrhea Cases from Children, Adults and Elderly at Northwest of Mexico.

Diarrheagenic Escherichia coli (DEC) strains are a main cause of gastrointestinal disease in developing countries. In this study we report the epidemiologic surveillance in a 4-year period (January 2011 to December 2014) of DEC strains causing acute diarrhea throughout the Sinaloa State, Mexico. DEC strains were isolated from outpatients of all ages with acute diarrhea (N = 1,037). Specific DEC pathotypes were identified by PCR-amplification of genes encoding virulence factors. The adhesion phenotype and antibiotic resistance were also investigated. DEC strains were detected in 23.3% (242/1037) of cases. The most frequently DEC strain isolated was EAEC [(12.2%), 126/242] followed by EPEC [(5.1%), 53/242], ETEC [(4.3%), 43/242] DAEC [(1.4%), 15/242], STEC [(0.3%), 3/242], and EIEC [(0.2%), 2/242]. EHEC strains were not detected. Overall DEC strains were more prevalent in children ≤2 years of age with EPEC strains the most common of DEC pathotypes. While ∼65% of EAEC strains were classified as typical variant based on the aggregative adherence to in vitro cultures of HEp-2 cells, a high proportion of EPEC strains was classified as atypical strains. EAEC, EPEC, ETEC, and DAEC strains were distributed in the north, central and south regions of Sinaloa state. Among all DEC strains, >90% were resistant to at least one commonly prescribed antibiotic. Strains were commonly resistant to first-line antibiotics such as tetracycline, ampicillin, and sulfamethoxazole-trimethoprim. Furthermore, more than 80% of DEC isolates were multi-drug resistant and EPEC and DAEC were the categories with major proportion of this feature. In conclusion, in nearly one out of four cases of acute diarrhea in Northwestern Mexico a multi-drug resistant DEC strain was isolated, in these cases EAEC was the most prevalent (52%) pathotype.

Nasopharyngeal Pneumococcal Density and Evolution of Acute Respiratory Illnesses in Young Children, Peru, 2009-2011.

We examined nasopharyngeal pneumococcal colonization density patterns surrounding acute respiratory illnesses (ARI) in young children in Peru. Pneumococcal densities were dynamic, gradually increasing leading up to an ARI, peaking during the ARI, and decreasing after the ARI. Rhinovirus co-infection was associated with higher pneumococcal densities.

The CpAL system regulates changes of the trans-epithelial resistance of human enterocytes during Clostridium perfringens type C infection.

Clostridium perfringens type C strains produce severe disease in humans and animals including enterotoxaemia and hemorrhagic diarrhea. Type C disease is mediated by production of toxins that damage the site of infection inducing loss of bloody fluids. Production of type C toxins, such as CPA, PFO, and, CPB is regulated by the C. perfringens Agr-like (CpAL) quorum sensing (QS) system. The CpAL system is also required to recapitulate, in vivo, intestinal signs of C. perfringens type C-induced disease, including hemorrhagic diarrhea and accumulation of fluids. The intestinal epithelium forms a physical barrier, made up of a series of intercellular junctions including tight junctions (TJs), adherens junctions (AJs) and desmosomes (DMs). This selective barrier regulates important physiological processes, including paracellular movement of ions and solutes, which, if altered, results in loss of fluids into the intestinal lumen. In this work, the effects of C. perfringens infection on the barrier function of intestinal epithelial cells was evaluated by measuring trans-epithelial resistance (TEER). Our studies demonstrate that infection of human enterocytes with C. perfringens type C strain CN3685 induced a significant drop on TEER. Changes in TEER were mediated by the CpAL system as a CN3685ΔagrB mutant did not induce such a drop. Physical contact between bacteria and enterocytes produced more pronounced changes in TEER and this phenomenon appeared also to be mediated by the CpAL system. Finally, immunofluorescence studies demonstrate that C. perfringens type C infection redistribute TJs protein occludin, and Claudin-3, and DMs protein desmoglein-2, but did not affect the AJs protein E-cadherin.

Bacterial Density, Serotype Distribution and Antibiotic Resistance of Pneumococcal Strains from the Nasopharynx of Peruvian Children Before and After Pneumococcal Conjugate Vaccine 7.

BACKGROUND: Pneumococcal conjugate vaccines (PCV) have decreased nasopharyngeal carriage of vaccine types but little data exist from rural areas. We investigated bacterial density, serotype distribution and antibiotic resistance of pneumococcal strains within the nasopharynx of young children in the Peruvian Andes, 2 years after PCV7 was introduced. METHODS: Pneumococcal strains were isolated from a subset of 125 children from our Peruvian cohort, who entered the study in 2009 and had pneumococcus detected in the nasopharynx in both 2009 and during follow-up in 2011. Strains were Quellung serotyped and tested for susceptibility to antibiotics. Bacterial density was determined by quantitative polymerase chain reaction. RESULTS: The prevalence of PCV7 strains decreased from 48% in 2009 to 28.8% in 2011, whereas non-PCV7 types increased from 52% to 71.2% (P = 0.002). There was a 3.5-fold increase in carriage of serotype 6C in 2011 (P = 0.026). Vaccination with PCV7 did not affect pneumococcal density in children colonized by a PCV7 type but did increase density in those colonized with a non-PCV7 type. Antibiotic resistance did not change after vaccine introduction; strains were nonsusceptible to tetracycline (97.2%), trimethoprim-sulfamethoxazole (56.4%), penicillin (34%), erythromycin (22.4%), chloramphenicol (18.8%) and clindamycin (12.4%). CONCLUSIONS: Serotype replacement was observed post-PCV7 vaccination with a concomitant, not previously recognized, increased nasopharyngeal density.

A pandemic Vibrio parahaemolyticus O3:K6 clone causing most associated diarrhea cases in the Pacific Northwest coast of Mexico.

Between September and October of 2004, more than 1230 cases of gastroenteritis due to pandemic O3:K6 strains of Vibrio parahaemolyticus (V. parahaemolyticus) were reported in the relatively small geographical area of Southern Sinaloa, a state located in Northwest Mexico. Since then, V. parahaemolyticus-associated gastroenteritis cases have gradually increased in prevalence spreading from south to north. The present study conducted an epidemiological surveillance of V. parahaemolyticus strains in both environmental and clinical samples along the Pacific coast of Sinaloa from 2011 to 2013. The genetic relatedness, serotype dominance and antibiotic resistance of isolates were investigated. A total of 46 strains were isolated from environmental samples (e.g., sediment, seawater and shrimp), whereas 249 strains were obtained from stools of patients with gastroenteritis. Nine different O serogroups and 16 serovars were identified. Serovars O3:K6 and O6:K46 were identified in both environmental and clinical strains. Whereas most environmental isolates carried the tdh gene (71.74%, 33/46), only three (6.52%) belonged to pandemic clones (O3:K6, O3:KUT and OUT:KUT). In contrast, 81.1% (202/249) of clinical isolates belonged to pandemic serotypes, with O3:K6 (tdh, toxRS/new, and/or orf8) representing the predominant serovar (97%, 196/202). This prevalence of pathogenic (tdh and/or trh positive) and O3:K6 pandemic V. parahaemolyticus isolates in this study were similar to those found from 2004 to 2010. As investigated by REP-PCR, genetic lineages of selected O3:K6 strains isolated in this study and some isolated earlier were nearly identical. Antimicrobial susceptibility testing showed that most strains (93.8%) were resistant to ampicillin but sensitive to chloramphenicol (98.8%). Multidrug resistance significantly increased from 8.6% (2004-2010) to 22.93% (2011-2013; p < 0.05). Our data indicate that pandemic O3:K6 clone has endemically established in the Pacific Coast of Mexico.

The role of influenza and parainfluenza infections in nasopharyngeal pneumococcal acquisition among young children.

BACKGROUND: Animal models suggest that influenza infection favors nasopharyngeal acquisition of pneumococci. We assessed this relationship with influenza and other respiratory viruses in young children. METHODS: A case-control study was nested within a prospective cohort study of acute respiratory illness (ARI) in Andean children <3 years of age (RESPIRA-PERU study). Weekly household visits were made to identify ARI and obtain nasal swabs for viral detection using real-time reverse-transcription polymerase chain reaction. Monthly nasopharyngeal (NP) samples were obtained to assess pneumococcal colonization. We determined whether specific respiratory viral ARI episodes occurring within the interval between NP samples increased the risk of NP acquisition of new pneumococcal serotypes. RESULTS: A total of 729 children contributed 2128 episodes of observation, including 681 pneumococcal acquisition episodes (new serotype, not detected in prior sample), 1029 nonacquisition episodes (no colonization or persistent colonization with the same serotype as the prior sample), and 418 indeterminate episodes. The risk of pneumococcal acquisition increased following influenza-ARI (adjusted odds ratio [AOR], 2.19; 95% confidence interval [CI], 1.02-4.69) and parainfluenza-ARI (AOR, 1.86; 95% CI, 1.15-3.01), when compared with episodes without ARI. Other viral infections (respiratory syncytial virus, human metapneumovirus, human rhinovirus, and adenovirus) were not associated with acquisition. CONCLUSIONS: Influenza and parainfluenza ARIs appeared to facilitate pneumococcal acquisition among young children. As acquisition increases the risk of pneumococcal diseases, these observations are pivotal in our attempts to prevent pneumococcal disease.

EspC promotes epithelial cell detachment by enteropathogenic Escherichia coli via sequential cleavages of a cytoskeletal protein and then focal adhesion proteins.

EspC is a non-locus of enterocyte effacement (LEE)-encoded autotransporter produced by enteropathogenic Escherichia coli (EPEC) that is secreted to the extracellular milieu by a type V secretion system and then translocated into epithelial cells by the type III secretion system. Here, we show that this efficient EspC delivery into the cell leads to a cytopathic effect characterized by cell rounding and cell detachment. Thus, EspC is the main protein involved in epithelial cell cytotoxicity detected during EPEC adhesion and pedestal formation assays. The cell detachment phenotype is triggered by cytoskeletal and focal adhesion disruption. EspC-producing EPEC is able to cleave fodrin, paxillin, and focal adhesion kinase (FAK), but these effects are not observed when cells are infected with an espC isogenic mutant. Recovery of these phenotypes by complementing the mutant with the espC gene but not with the espC gene mutated in the serine protease motif highlights the role of the protease activity of EspC in the cell detachment phenotype. In vitro assays using purified proteins showed that EspC, but not EspC with an S256I substitution [EspCS256I], directly cleaves these cytoskeletal and focal adhesion proteins. Kinetics of protein degradation indicated that EspC-producing EPEC first cleaves fodrin (within the 11th and 9th repetitive units at the Q1219 and D938 residues, respectively), and this event sequentially triggers paxillin degradation, FAK dephosphorylation, and FAK degradation. Thus, cytoskeletal and focal adhesion protein cleavage leads to the cell rounding and cell detachment promoted by EspC-producing EPEC.

Cohort profile: The study of respiratory pathogens in Andean children.

We investigated respiratory pathogens in a prospective cohort study of young children living in the Peruvian Andes. In the study we assessed viral respiratory infections among young children, and explored interactions of viruses with common respiratory bacteria, especially Streptococcus pneumoniae. Through weekly household visits, data were collected on the signs and symptoms of acute respiratory illness (ARI), nasal samples were collected to test for viruses during episodes of ARI, and nasopharyngeal samples were collected on a monthly basis to monitor bacterial colonisation. We also collected data on vaccination coverage, patterns of social mixing, geographic information, and environmental and socio-demographic variables. Understanding the interaction of respiratory viruses with bacteria and its impact on the burden and severity of ARIs in rural areas of developing countries is critical to designing strategies for preventing such infections.