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Diarrheagenic Escherichia coli is one of the most prevalent foodborne pathogens and poses significant clinical challenges due to increasing rates of antibiotic resistance. In this study, 9,068 diarrheagenic E. coli isolates from sporadic diarrhea cases across 20 provinces of China from 2016 to 2022 were systematically analyzed to determine the prevalence and distribution of resistance. A total of 14 antibiotics were included in the antimicrobial resistance surveillance. Antimicrobial susceptibility testing indicated that the majority of the isolates (89.3%) were resistant to at least one antimicrobial agent, with a significant proportion (60.3%) demonstrating multidrug resistance. Isolates showed high levels of resistance to ampicillin (68.2%) and high sensitivity to imipenem (97.0%). Over the 7 years, an increasing trend in resistance to cefoxitin and chloramphenicol among the isolates was found. Enteroaggregative E. coli was identified as the predominant pathotype. Furthermore, antimicrobial resistance rates were higher among the isolates from infants under 1 year of age compared to other populations and resistance levels differed among pathotypes. In summary, our findings revealed the detailed resistance characteristics of diarrheagenic E. coli strains isolated from human sporadic diarrhea cases in China, offering valuable insights that can inform clinical antibiotic treatment strategies and priorities. [ABSTRACT FROM AUTHOR]

The diarrheagenic Escherichia coli (DEC) is the major cause of diarrheal diseases in Africa, including Ethiopia. However, the genetic diversity of E. coli pathotypes found in Ethiopia has not been studied well. This study aimed to characterize potential DEC belonging to enteropathogenic (EPEC), Shiga toxin-producing (STEC), enteroaggregative (EAEC), enterotoxigenic (ETEC), and enteroinvasive (EIEC) E. coli pathotypes from stool specimens of patients attending primary healthcare units (n = 260) in Addis Ababa and Hossana using whole-genome sequencing. Real-time PCR assays were used to identify DEC isolates belonging to EPEC, STEC, EAEC, ETEC, and EIEC pathotypes, which were then subjected to whole-genome sequencing on the Illumina platform. Twenty-four whole-genome nucleotide sequences of DEC strains with good enough quality were analyzed for virulence-associated genes (VAGs), antibiotic resistance genes (ARGs), phylogenetic groups, serogroups, and sequence types. The majority (62.5%) of DEC isolates belonged to the phylogenetic group B1. The identified DEC isolates belonged to 21 different serogroups and 17 different sequence types. All tested DEC isolates carried multiple VAGs and ARGs. The findings highlight the high diversity in the population structure of the studied DEC isolates, which is important for designing targeted interventions to reduce the diarrheal burden in Ethiopia. [ABSTRACT FROM AUTHOR]

The list of standard abbreviations for JDS is available at adsa.org/jds-abbreviations-24. Nonstandard abbreviations are available in the Notes. Diarrheagenic Escherichia coli (DEC) is a kind of foodborne pathogen that poses a significant threat to both food safety and human health. To address the current challenges of high prevalence and difficult subtyping of DEC, this study developed a method that combined multiplex PCR with high-resolution melting (HRM) analysis for subtyping 5 kinds of DEC. The target genes are amplified by multiplex PCR in a single well, and HRM curve analysis was applied for distinct amplicons based on different melting temperature (Tm) values. The method enables discrimination of different DEC types based on characteristic peaks and distinct Tm values in the thermal melting curve. The assay exhibited 100% sensitivity and 100% specificity with a detection limit of 0.5 to 1 ng/μL. The results showed that different DNA concentrations did not influence the subtyping results, demonstrating this method owed high reliability and stability. In addition, the method was also used for the detection and subtyping of DEC in milk. This method streamlines operational procedures, shorts the detection time, and offers a novel tool for subtyping DEC. [ABSTRACT FROM AUTHOR]

Background: Diarrheagenic Escherichia coli (DEC) infections constitute the leading causes of morbidity and mortality among children in Sub-Saharan Africa. However, little has so far been done to properly reveal the pathogenic endowments of DEC in these populations. Aims and Objectives: We evaluated 4 DEC strains among children under 5 years. Materials and Methods: A cross-sectional study design was employed among 384 positive cases. Results: There was a significant decline in infections associated with DEC as the children grew older (χ²[12] = 87.366: P = [0.000]. A total of 56 (14.6%) cases were 0-12 months, 168 (43.8%) were 13-24 months, 88 (22.9%) were 25-36 months, 40 (10.4%) were 37-48 months, and 32 (8.3%) were 49-60 months. A total of 248 (64.6%) male subjects exhibited more susceptibility to DEC infections than their female counterparts (n = 136 [35.4%]) (χ² [3] =13.313: P = [0.004]. Subjects from urban areas (n = 248 [64.6%]), significantly bored the brunt of infections than those from rural areas (n = 136 [35.4%]) (χ² [3] = 35.147: P = [0.000]. The prevalence of DEC appeared significantly higher during rainy seasons (n = 269 [70.1%]). Conclusion: Young age, male gender, crowding, and rainy season play a central role in the transmission of DEC pathotypes. [ABSTRACT FROM AUTHOR]

The safety of fecal microbiota transplantation (FMT) has been highlighted by extended-spectrum β-lactamase (ESBL)-producing Escherichia coli bacteremia transmitted from donors and acquisition of diarrheagenic E. coli (Shiga toxin-producing E. coli (STEC) and enteropathogenic E. coli (EPEC)) via FMT. The use of donor screening criteria to lower the risk of pathogen transmission via FMT is essential. This study aimed to demonstrate the outcomes of our strict donor screening program. This study was conducted at our FMT center between January 2019 and June 2022. Our donor screening program included an initial questionnaire and subsequent blood and stool testing. We further used selective culture for third-generation cephalosporin-resistant (3GCR) Enterobacterales and multiplex PCR to detect diarrheagenic E. coli in stools. The resistance mechanisms and sequence type of 3GCR Enterobacterales were determined. A total of 742 individuals were assessed, and 583 participants (78.6%) were excluded after questionnaire. Of the remaining 159 participants undergoing stool and blood tests, 37 participants were finally qualified (5.0%, 37/742). A high fecal carriage rate of ESBL-producing Enterobacterales (35.2%, 56/159), including E. coli (n=53) and Klebsiella pneumoniae (n=5), and diarrheagenic E. coli (31.4%, 50/159), including EPEC (n=41), enteroaggregative E. coli (n=11), enterotoxigenic E. coli (n=4), and STEC (n=1), was noted. CTX-M-79 and CTX-M-15 were dominant in E. coli and K. pneumoniae, respectively. The sequence types of the ESBL-producing strains were diverse. The screening for 3GCR Enterobacterales and diarrheagenic E. coli in stool is necessary. Our findings also support the effectiveness of multiplex PCR panels in FMT donor screening programs. [ABSTRACT FROM AUTHOR]

Introduction: Diarrheal diseases are common with worldwide distribution, and diarrheagenic Escherichia coli (DEC) strains are the main causative agents. The present study aimed to define the association of various pathotypes of E. coli from diarrheal patients in Mongolia. Methodology: A total of 341 E. coli strains were isolated from the stool of diarrheal patients. Bacterial susceptibility to antimicrobial agents was determined by the Kirby Bauer disk diffusion method. DEC isolates were identified by HEp-2 cell adherence assay and multiplex polymerase chain reaction (PCR). Results: DEC pathogens were detected in 53.7% of 341 E. coli isolates. Enteroaggregative E. coli (EAEC) was the most common DEC pathotype identified by HEp-2 adherence assay and multiplex PCR methods in 97 samples (28.4%), followed by atypical enteropathogenic E. coli (EPEC) in 50 samples (14.7%), diffusely adherent E. coli (DAEC) in 25 samples (7.3%), enterohaemorrhagic E. coli (EHEC) in 6 samples (1.8%), enterotoxigenic E. coli (ETEC) in 4 samples (1.2%), and enteroinvasive E. coli (EIEC) in 1 sample (0.3%). DEC strains had > 50% antibiotic resistance against cephalothin, ampicillin, and trimethoprim/sulfamethoxazole. All tested DEC strains were susceptible to imipenem. Among the 183 DEC strains, 27 (14.8%) were extended spectrum beta-lactamase producing isolates, and 125 (68.3%) isolates were multiple drug resistant. Conclusions: We have identified six pathotypes of DEC from the clinical isolates tested and concluded that a high prevalence of antimicrobial resistance was observed in these pathotypes. EAEC was the most common pathotype identified and this is the first report of EHEC identification in Mongolia. [ABSTRACT FROM AUTHOR]

The current study aimed to detect virulence, hetero-pathogenicity, and hybridization genes in Escherichia coli strains, previously isolated from cloacal swabs in commercial breeding psittacines and zoological collections, via multiplex PCR. A total of 68 strains of E. coli , previously isolated from psittacines in zoos and commercial breeding facilities in Ceará, Brazil, were assessed for the presence of the following genes and/or probes: eae , bfpA (EPEC - Enteropathogenic E. coli), CVD432 (EAEC - Enteroaggregative E. coli); LT gene and ST gene (ETEC - Enterotoxigenic E. coli); ipaH (EIEC - Enteroinvasive E. coli); stx1 and stx2 (STEC - Shiga toxin-producing E. coli); iroN , ompT , hlyF , iss , and iutA (APEC - Avian pathogenic E. coli). Of the 68 E. coli strains analyzed, 61 (98.7 %) were positive for the following genes and/or probes: Stx1 (61/98.7 %), ST gene (54/79.4 %), CVD432 (49/72 %), bfpA (44/64.7 %), eae (42/61.8 %), Stx2 (41/60.3 %), ipaH (34/50 %), LT gene (33/48.5 %), iroN (21/30.9 %), hlyF (11/6.2 %), iss (06/8.8 %) and iutA (06/8.8 %). The following diarrheagenic pathotypes were identified: 66 (97 %) from STEC, 49 (72 %) from EAEC, 35 (52 %) from EIEC, 25 (37 %) from ETEC, and one (1.5 %) from EPEC. Regarding hetero-pathogenicity, 50 (74 %) heterogeneous strains were identified. Positivity for APEC was seen in four (6 %) strains, all characterized as pathogenic hybrids. This study describes significant associations of virulence factors in E. coli strains DEC/DEC and DEC/APEC, which were isolated from psittacines and may be potentially harmful to One Health. • Psittacines under human care can carry potentially zoonotic pathogens. • Diarrheagenic and avian pathogenic Escherichia coli can share genes and vice versa. • Virulence factors in E. coli highlight the importance of effective sanitary control. [ABSTRACT FROM AUTHOR]

Introduction Diarrheal illness such as diarrheagenic Escherichia coli (DEC), apart from rotavirus, is a common etiological agent known to cause moderate-to-severe diarrhea in low-income countries where unregulated use of antibiotics is rampant, giving rise to multidrug resistant (MDR) strains. This study is an earnest effort in reflecting the resistance pattern in such isolates. Materials and Methods It is a hospital-based cross-sectional study conducted over a period of 1 year (January to December, 2015). Children aged less than 18 years presenting with (n = 170) and without (n = 47) diarrhea were included as cases and controls, respectively. Fresh stool sample from eligible participants was collected and inoculated on MacConkey agar. Based on the colony morphology and biochemical identification followed by polymerase chain reaction (PCR), different pathotypes of DEC were identified. All such isolates were subjected to antimicrobial susceptibility testing employing VITEK 2 identification system. The result of the tested antibiotics was evaluated as per Clinical and Laboratory Standards Institute 2015 guidelines. Results DEC with specific virulence genes were detected by multiplex real-time PCR in 39 and 3 children with or without diarrhea, respectively. Most common DEC pathotypes found were enteroaggregative E. coli (38%) followed by enteropathogenic E. coli (28.5%). MDR isolates comprised 35 of 42 DEC pathotypes (83.3%). Resistance among DEC pathotypes to ampicillin, amoxicillin–clavulanate, ciprofloxacin, cephalosporin, nalidixic acid, imipenem, and cotrimoxazole was found to be statistically significant in comparison to non-DEC isolates. Conclusion This study has highlighted the increased prevalence of MDR strains among DEC pathotypes. Looking for these isolates will help detect dreadful DEC pathotypes like enterohemorrhagic E. coli where early administration of a sensitive antibiotic will go a long way in preventing complication like hemorrhagic colitis and hemolytic uremic syndrome. [ABSTRACT FROM AUTHOR]

Background and Objectives: Pathogenic diseases resulting from microbial contamination of food have been widely distributed in many parts of the world. Among these, Escherichia coli is one of the most important foodborne pathogenic bacteria. Diarrhea is one of the major causes of children's death in developing countries, with approximately 2 million deaths annually. The current study aimed to determine the frequency of diarrheagenic E. coli pathotypes such as Enteropathogenic E. coli (EPEC), Enterotoxigenic E. coli (ETEC), Enteroaggregative E. coli (EAEC), and Shiga toxin-producing E. coli (STEC) in Brassica oleracea cultivars in order to provide information on the assessment of diarrheagenic E. coli pathogenesis risk. Materials and Methods: 100 samples of vegetables were collected in Tehran, including cabbage, cauliflower, broccoli and Brussels sprouts. After homogenizing samples, enrichment was done in the EC broth medium. Five colonies of pure culture were used for DNA extraction. Pathotypes were identified by PCR using virulence genes. Results: The results showed that the prevalence of diarrheagenic E. coli strains was 7%. The EPEC prevalence was 3%, All EPEC isolates were atypical. The ETEC frequency was 3%, And the EAEC prevalence was 1%. Conclusion: These findingsindicated that Brassica oleracea cultivars could be consideredas a source of contamination with diarrhea-causing E. coli strains. [ABSTRACT FROM AUTHOR]

Pathogenic bacteria, viruses, and parasites can cause waterborne disease outbreaks. The study of coastal water quality contributes to identifying potential risks to human health and to improving water management practices. The Río de la Plata River, a wide estuary in South America, is used for recreational activities, as a water source for consumption and as a site for sewage discharges. In the present study, as the first step of a quantitative microbial risk assessment of the coastal water quality of this river, a descriptive study was performed to identify the microbial pathogens prevalent in its waters and in the sewage discharged into the river. Two sites, representing two different potential risk scenarios, were chosen: a heavily polluted beach and an apparently safe beach. Conductivity and fecal contamination indicators including enterococci, Escherichia coli , F + RNA bacteriophages, and human polyomaviruses showed high levels. Regarding enterococci, differences between sites were significant (p-values <0.001). 93.3% and 56.5% of the apparently safe beach exceeded the recreational water limits for E. coli and enterococci. Regarding pathogens, diarrheagenic E. coli , Salmonella , and noroviruses were detected with different frequencies between sites. The parasites Cryptosporidium spp. and Giardia duodenalis were frequently detected in both sites. The results regarding viral, bacterial, and parasitic pathogens, even without correlation with conventional indicators, showed the importance of monitoring a variety of microorganisms to determine water quality more reliably and accurately, and to facilitate further studies of health risk assessment. The taxonomic description of microbial pathogens in river waters allow identifying the microorganisms that infect the population living on its shores but also pathogens not previously reported by the clinical surveillance system. • Detection of viral, bacterial and parasitic pathogens in recreational river waters. • Bacterial and viral pathogens, but not parasites, decreased with distance from sewage discharges. • The taxonomy of environmental pathogens agreed with that of clinical cases. • Environmental surveillance identified pathogens not yet reported in clinical surveys. • Monitoring of a variety of microorganisms for a reliable and accurate determination of water quality. [ABSTRACT FROM AUTHOR]

Background and Objective: Escherichia coli pathogenic strains are considered as the main cause of intestinal and extra-intestinal diseases. This study aimed to identify and evaluate the dominant pathotypes of diarrheagenic E. coli isolates among children sufering from diarrhea in Khoram Abad city, Iran. Subjects and Methods: In this current study, 578 diarrhea samples of the children less than 10 years old were collected in Khoram Abbad city. After culture and isolation, the pathogenic groups of diarrheagenic E. coli (DEC) isolates including four pathotypes of Enterotoxigenic E. coli, Enteropathogenic E. coli, Enteroinvasive E. coli, and Shiga toxigenic E. coli were investigated by PCR. Disk diffusion method (Kirby Bauer) was used for detection of antimicrobial sensitivity of the isolates. Results: Out of 578 stool samples collected 81 isolates were DEC. The frequencies of the isolates were respectively: Enterotoxigenic E. coli (15.1%), enteropathogenic E. coli (14%), Enteroinvasive E. coli (5%), and Shiga toxigenic E. coli (9.7%). The highest antibiotic resistance levels were respectively reported to: amoxicilin (78%), tetracycline (74.1%), co-trimoxazole (64.2%), nalidixic acid (56.8%), streptomycin (46.9%), minocycline (38.3%), and ciprofloxacin (16.1%). Multiple drug resistance was found in 30.9% of the isolates. Conclusion: Enterotoxigenic E. coli is one of the most common diarrheagenic agent affecting the children less than 10 years old in this region. Therefore, such diseases are preventable by controlling water and food hygiene especially in warm seasons. [ABSTRACT FROM AUTHOR]

Diarrheagenic Escherichia coli (DEC) is a group of important foodborne pathogens that can spread and cause infection in humans. As a significant public health issue, DEC infection may lead to a series of diseases such as abdominal pain, diarrhea, hemorrhagic colitis, and hemolytic uremic syndrome. Therefore, its rapid and accurate detection has become an important concern for food safety issues and healthcare-associated infections. We have developed a multiplex real-time quantitative PCR assay (qPCR), DEC Sensor Test, to simultaneously recognize 5 major DEC pathotypes in a single tube, enterotoxigenic, enterohemorrhagic, enteropathogenic, enteroaggregative and enteroinvasive E. coli. In this assay, 13 DEC virulence genes, stp, sth, lt, stx1, stx2, eae, escV, bfpB, aggR, astA, pic, invE, and ipaH, can be simultaneously detected along with a spiked-in internal amplification control to avoid false negative results. All the constituent PCR for the 13 DEC virulence genes had high analytical sensitivity and specificity. Detection of all virulence genes by DEC Sensor Test can be much easier, cheaper and faster to perform than conventional tests. It also has added advantages of being able to distinguish live or dead bacteria as well as eliminating the possibility of switching partial results between different samples associated with multi-tube tests. Therefore, the application of this assay may significantly enhance our ability to prevent and fight DEC-associated diseases and to improve public health. [ABSTRACT FROM AUTHOR]

Aim: This article aimed to analyze the diarrheagenic potential of E. coli isolated from urinary tract infection (UTI) and to recognize the presence of antibiotic resistance genes. Background: The marked genome plasticity of Escherichia coli has allowed the emergence of resistant pathogenic strains displaying an unusual arrangement of genes. Methods: In this cross-sectional study, 110 E. coli were isolated from patients with the symptoms of UTI in Sanandaj, west of Iran between July and September - 2015. The isolates were examined by the disk diffusion method for antibiotic susceptibility test and by polymerase chain reaction for the presence of genes characteristic of diarrheagenic E. coli (DEC), Uropathogenic E. coli (UPEC) virulence genes, extended-spectrum β-lactamase blaCTX-M and plasmid-mediated quinolone resistance determinants, qnrA, qnrB, and qnrS. Results: The most and the least effective antibiotics were nitrofurantoin and cefotaxime (96.4% and 27.3% sensitivity, respectively). Of the 110 UTI isolates, 57.3% carried diarrheagenic genes. The bundle-forming pilus bfpA was the most prevalent diarrheagenic gene (39.1%). The most commonly detected DEC pathotype was enterotoxigenic E. coli (-ETEC, 12.7%). All the pathotypes carried the blaCTX-M and qnr. The -UPEC hly hemolysin and pap adhesin genes were mainly detected among ETEC isolates Conclusion: Our results indicated the presence of resistant diarrheagenic pathotypes in UTI-associated E. coli. Such isolates may have the capacity of causing both extraintestinal and intestinal infections. Based on our knowledge, this is the first report of the presence of qnr in ETEC from urine. [ABSTRACT FROM AUTHOR]

Diarrheagenic Escherichia coli (DEC) is a leading cause of foodborne illness associated with intestinal disease. While known over the years that contamination of food sources occurs via the oral faecal-route, the mechanisms underlying its persistence within the open environments including the food chain remains virtually unknown. Therefore, in this mini-review we will shed light on bacterial processes such as initial attachment, biofilm formation, horizontal gene transfer and response to environmental stresses. These factors may enable persistence of DEC as well as the emergence of potentially more virulent strains within the agricultural and food production environment. Mechanistic studies in clinical microbiology and immunology have elucidated infection pathways in the human and other animal bodies leading to diagnostic and treatment solutions. Therefore, understanding DEC behaviour in the agricultural and food production environment is crucial for ensuring food safety and public health by reducing the burden of foodborne illnesses. • Diarrheagenic Escherichia coli (DEC) is a leading cause of foodborne outbreaks worldwide. • DEC's ability to survive within the open environment has been scarcely explored. • DEC characteristics may aid survival in open the environment subsequently contaminating food sources. • Bacterial adherence, lateral gene transfer and environmental response may persistence of DEC persistence. • These factors elucidate how foodborne pathogens spread along the food chain. [ABSTRACT FROM AUTHOR]

INTRODUCTION: Infectious diarrhea is leading infectious cause of childhood morbidity, hospitalizations, and mortality particularly in children living in developing countries like India. The etiological agents differ depending on geographical area, and recent data suggest increase in drug resistance to various enteropathogens. AIMS AND OBJECTIVES: The aim of the study was to investigate emerging diarrheal agents and antimicrobial resistance profile of bacterial pathogens from children (<12 years of age) hospitalized with acute diarrhea. MATERIALS AND METHODS: A cross-sectional, hospital-based observational study was conducted over 1 year in which 100 children <12 years who were hospitalized due to diarrhea were recruited. Diarrhea was defined as the passage of three or more liquid stools in a 24-h period using the World Health Organization guidelines. Samples were processed for detection of various bacterial, viral, and parasitic agents by standard microbiological, serological, and molecular tests. Antimicrobial resistance testing was performed with the Kirby-Bauer disk diffusion method. ELISA was performed for Rotavirus and Escherichia coli O157. Multiplex polymerase chain reaction test was performed to detect diarrheagenic E. coli (DEC). RESULTS: Pathogenic diarrheal agents were found in 63% patients. Rotavirus was identified in 52.5%, DEC in 29%, Vibrio cholerae in 4%, Shigella flexneri in 3%, Aeromonas sp. in 1%, Giardia lamblia in 4%, and Entamoeba histolytica in 1% cases. Enteropathogenic E. coli (EPEC) in 19 (65.5%) cases was the most common agent followed by Enteroaggregative E. coli (EAEC) in 5 (17.2%), Enterotoxigenic E. coli (ETEC) in 2 (6%), and Enteroinvasive E. coli (EIEC) in 3 (10.3%) cases. Resistance rates of DEC to first-line therapeutic drugs were high, 97.3% to ampicillin and 95.95% to co-trimoxazole. DEC was susceptible to chloramphenicol in 58.11%, gentamicin in 48.19%, and amikacin in 58.11% cases. Shigella sp. and V. cholerae isolates were 100% sensitive to gentamicin and ofloxacin. CONCLUSION: EPEC is the most common DEC pathotype and EAEC, ETEC, and EIEC are also emerging as dominant diarrheal agents. Rotavirus was the most common causative agents of diarrhea especially in children <5 years. Most of the bacterial isolates showed high level of drug resistance to first-line empirical drugs and were multidrug resistant making them unsuitable for empiric treatment. Laboratory monitoring of drug susceptibility of stool isolates appears necessary to formulate antibiotic policy for treating diarrheal illness at the local level. There is an urgent need to strengthen diarrheal surveillance to monitor susceptibility to commonly prescribed antibiotics. [ABSTRACT FROM AUTHOR]

Human movement may be an important driver of transmission dynamics for enteric pathogens but has largely been underappreciated except for international 'travelers' diarrhea or cholera. Phylodynamic methods, which combine genomic and epidemiological data, are used to examine rates and dynamics of disease matching underlying evolutionary history and biogeographic distributions, but these methods often are not applied to enteric bacterial pathogens. We used phylodynamics to explore the phylogeographic and evolutionary patterns of diarrheagenic E. coli in northern Ecuador to investigate the role of human travel in the geographic distribution of strains across the country. Using whole genome sequences of diarrheagenic E. coli isolates, we built a core genome phylogeny, reconstructed discrete ancestral states across urban and rural sites, and estimated migration rates between E. coli populations. We found minimal structuring based on site locations, urban vs. rural locality, pathotype, or clinical status. Ancestral states of phylogenomic nodes and tips were inferred to have 51% urban ancestry and 49% rural ancestry. Lack of structuring by location or pathotype E. coli isolates imply highly connected communities and extensive sharing of genomic characteristics across isolates. Using an approximate structured coalescent model, we estimated rates of migration among circulating isolates were 6.7 times larger for urban towards rural populations compared to rural towards urban populations. This suggests increased inferred migration rates of diarrheagenic E. coli from urban populations towards rural populations. Our results indicate that investments in water and sanitation prevention in urban areas could limit the spread of enteric bacterial pathogens among rural populations. • We explore impact of human mobility on the spread of diarrheagenic E. coli across urban-rural communities. • Minimal structuring of circulating diarrheagenic E. coli core genomes based on key metadata suggest extensive regional mixing. • Estimated rates of migration among diarrheagenic E. coli were larger for urban towards rural populations. • Our approach contextualizes broader drivers of diarrheal disease transmission that can complement traditional WASH efforts. [ABSTRACT FROM AUTHOR]

Introduction: Diarrheagenic Escherichia coli (DEC) is a major etiologic agent among the pathogens that cause diarrhea in children. Methodology: To investigate the presence and pathotypes of DEC in children under five years of age, living in the province of Khouzestan, Iran. 208 diarrhea stool samples were screened by multiplex-PCR. The isolated DEC isolates were investigated for resistance to various antimicrobials including the production of extended-spectrum beta-lactamases (ESBLs) and phylogenetic groups were determined. Results: DEC isolates were identified in 54 (26%) diarrhea samples, and 4 (7%) cases contained two DEC pathotypes. DEC isolated included 35 (16.8%) enteroaggregative E. coli (EAEC), ten (4.8%) enteropathogenic E. coli (EPEC), six (2.9%) enteroinvasive E. coli (EIEC), six (2.9%) enterotoxigenic E. coli (ETEC) and one (0.48%) LEE-positive EAEC. Shiga-toxin producing E. coli (STEC) was not identified in any diarrheal samples. The most prevalent resistance was observed with ceftazidime (88%), followed by ceftizoxime (83%) and ceftriaxone (71%). The majority of isolates (> 75%) were sensitive to Imipenem, ciprofloxacin, and amikacin. More than 65% of the pathogenic isolates showed a multidrug-resistant phenotype. ESBL-producing strains was observed in 79.3% of all DEC isolates. Phylogenetic group B2 was the most predominant group with a frequency of 44.8%. A significant association was observed between the B2 phylogenetic group and the DEC isolates (P < 0.05). Conclusions: Overall, our findings highlight the importance of the role of DEC isolates in the etiology of diarrhea in children in Iran. The progressive increase in antimicrobial resistance among DEC isolates makes it imperative to implement policies to control the spread of resistant bacteria. [ABSTRACT FROM AUTHOR]

Background: Diarrhea is the leading infectious cause of childhood morbidity and mortality. Among bacterial agents, diarrheagenic Escherichia coli (DEC) is the major causal agent of childhood diarrhea in developing countries, particularly in children under the age of 5 years. Here, we performed a hospital-based prospective study to explore the pathotype distribution, epidemiological characteristics and antibiotic resistance patterns of DEC from < 5-year-old diarrheal children.Methods: Between August 2015 and September 2016, 684 stool samples were collected from children (< 5 years old) with acute diarrhea. All samples were cultured and identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) and biochemical tests. PCR was used for subtyping, and enteropathogenic E. coli (EPEC) isolates were identified simultaneously with serology. Furthermore, antimicrobial sensitivity tests and sequencing of antibiotic resistance-related genes were conducted.Results: DEC strains were identified in 7.9% of the 684 stool samples. Among them, the most commonly detected pathotype was EPEC (50.0% of DEC), of which 77.8% were classified as atypical EPEC (aEPEC). Age and seasonal distribution revealed that DEC tended to infect younger children and to occur in summer/autumn periods. Multidrug-resistant DEC isolates were 66.7%; resistance rates to ampicillin, co-trimoxazole, cefazolin, cefuroxime, cefotaxime, and ciprofloxacin were ≥ 50%. Among 5 carbapenem-resistant DEC, 60.0% were positive for carbapenemase genes (2 blaNDM-1 and 1 blaKPC-2). Among 30 cephalosporin-resistant DEC, 93.3% were positive for extended-spectrum β-lactamase (ESBL) genes, with blaTEM-1 and blaCTX-M-55 being the most common types. However, no gyrA or gyrB genes were detected in 16 quinolone-resistant isolates. Notably, aEPEC, which has not received much attention before, also exhibited high rates of drug resistance (81.0%, 66.7%, and 14.3% for ampicillin, co-trimoxazole , and carbapenem resistance, respectively).Conclusions: EPEC was the most frequent DEC pathotype in acute diarrheal children, with aEPEC emerging as a dominant diarrheal agent in central China. Most DEC strains were multidrug-resistant, making even ciprofloxacin unsuitable for empiric treatment against DEC infection. Among carbapenem-resistant DEC strains, those harboring blaNDM-1 and blaKPC-2 were the main causal agents. blaTEM-1 and blaCTX-M-55 were the major genetic determinants associated with high levels of cephalosporin resistance. [ABSTRACT FROM AUTHOR]

Background The population structure of human diarrheagenic Escherichia coli (DEC) isolates derived from worldwide collections remains undefined. Methods A total of 1196 clinical isolates were obtained from a multilocus sequence typing (MLST) database. Genetic diversity analysis, MLST analysis, and phylogenetic analysis combined with different pathotypes were performed through a variety of calculation software applications. Results All isolates were categorized as one of 579 different sequence types (STs). The eBURST algorithm resolved these 579 STs into 27 clonal complexes (CCs), 37 concatemers, and 210 singletons, revealing a high level of genetic diversity in the population structure of DEC. CC10 was the most prevalent CC, comprising 276 (23.08%, 276/1196) isolates with 85 (14.68%, 85/579) STs widely distributed in 20 countries. The population structure of five common pathotypes was highly diversified, and isolates with the same ST or CC were heterogeneous for different pathotypes. Sequence variations were more abundant in fumC and gyrB than in the other five genes, and these exhibited the highest degree of nucleotide diversity (0.03886 and 0.03075, respectively) and the greatest number of polymorphic nucleotide sites (137 and 139, respectively). The dN/dS ratios of seven analyzed loci varied from 0.0083 ( recA ) to 0.0434 ( purA ), and the ratio for the concatenated sequence was 0.2518, revealing the effects of purifying selection on housekeeping genes during the evolutionary process. Significant allele linkage disequilibrium was detected when the standardized index of association ( I S A ) was calculated both for the entire collection of isolates (0.3174, p < 0.001) and for the 579 STs (0.1475, p < 0.001). Conclusions This study facilitated a comprehensive understanding of the genetic diversity of human DEC distributed across the global population. The results provide genetic evidence that will allow us to uncover the microevolutionary relationships among different pathogenic isolates of DEC. [ABSTRACT FROM AUTHOR]

Diarrheagenic E. coli (DEC) isolates were recovered from local retail markets and the Osaka Municipal Central Wholesale Market in Japan. Retail food samples were collected for analysis in Osaka Japan from 2005 to 2008 and consisted of 32 beef, 28 pork, 20 poultry, 136 fish, 66 fruits and vegetables and 51 ready–to-eat (RTE) food samples. A total of 82 DEC strains were recovered from 64 (19%) food samples with the highest prevalence in poultry (100%, 20/20), followed by pork (54%, 15/28), beef (28%, 9/32), fruits and vegetables (12%, 8/66), fish (6.6%, 9/136) and RTE foods (5.9%, 3/51). Most of the strains belonged to E. coli possessing the enteroaggregative E. coli (EAEC) heat-stable enterotoxin 1 (EAST1) gene (EAST1EC; n = 62, P < 0.0001) and enteropathogenic E. coli (EPEC; n = 16, P < 0.01), whereas only 1 strain belonged to Shiga toxin-producing E. coli (STEC), 1 to EAEC and 2 to enterotoxigenic E. coli (ETEC) strains. Of the 82 DEC isolates, 22 O and 13H serogroups were detected, including some specific serogroups (O91, O103, O115, O119, O126, and O157) which have been associated with human diarrheal infections. Phylogenetic group A and B1 were predominant among the DEC isolates. Antimicrobial resistance to tetracycline was most common (49%), followed by nalidixic acid (28%), ampicillin (24%), sulfamethoxazole/trimethoprim (20%), and cephalothin (18%). All isolates were susceptible to aztreonam. Of the resistant strains, 44% (22/50) demonstrated resistance to > 3 antimicrobial agents. Isolates resistant to > 5 antimicrobials were only found in the meat samples, while isolates from the fruits and vegetables as well as RTE foods showed resistance to only 1 or 2 antimicrobial agents. Sixty one percent of EAST1EC, 56% of EPEC and all of the EAEC and ETEC were resistant to at least 1 antimicrobial agent. Multiple-locus variable-number tandem repeat analysis (MLVA) was used in this study for genotyping of DEC. The 82 isolates collected for this study showed 77 distinct MLVA profiles located among 3 branches. The Simpson's Index of Diversity ( D ) was 99.9% at its highest. The high diversity of these food strains would suggest their originating from a variety of sources and environments. In conclusion, retail food samples in Japan were contaminated with DEC; EAST1EC, a putative DEC, were detected at high rates in poultry, pork and beef. Isolates resistant to > 3 antimicrobials were found only in raw meat and fish. Food animals may act as the reservoir for multi-resistant bacteria. Due to the finding that nearly 1/3 of EAST1EC strains were resistant to > 3 antimicrobials, additional surveillance for EAST1EC should be initiated. [ABSTRACT FROM AUTHOR]

This study aimed to evaluate the profile of antimicrobial susceptibility and the occurrence of diarrheagenic pathotypes (E. coli entoropatogênic/EPEC, E. coli enterotoxigenic/ETEC, E. coli producing shiga toxin/STEC, E. coli enteroinvasive/EIEC and E. coli enteroaggregative/EAEC) in Escherichia coli isolated from different foods intended for human consumption and those involved in foodborne disease outbreaks, in Minas Gerais, Brazil. The antimicrobial susceptibility of strains of E. coli was evaluated by the Minimum Inhibitory Concentration technique and pathotypes of E. coli diarrheagenic by polymerase chain reaction, using the virulence markers eae, bfpA, eltB, st1, stx1, stx2, ipaH and aatA. Among the 220 strains of E. coli analyzed, 60 (27.3%) were resistant or intermediate resistance. Among these, eight (13.3%) were considered multiresistant. The amikacin and gentamicin drugs were the most effective, unlike ampicillin, which had the highest percentage of resistant strains (10.9%). Pathotypes of EPEC, STEC, EIEC and EAEC were not identified. Two strains of E. coli (0.9%) isolated from a salty (cheese bread) and a mixed meal (chicken meal) were classified as ETEC. Both harbored the virulence markers eltB, estA and st1. These were characterized as serotypes O9:H10 and O9:H33. We conclude that among samples of E. coli, isolated from foods for human consumption in Minas Gerais and those involved in outbreaks of DTA, there are samples considered multiresistant and diarrheogenic. [ABSTRACT FROM AUTHOR]

Microbiological surveillance of ready-to-eat (RTE) meat products provides empirical data to inform scientific guidance for improving food safety. In this study we conducted the first nation-wide survey of food-borne pathogens in RTE meat products in China in 2013. We analyzed the prevalence of food-borne pathogen in different product categories, market distribution, packaged forms, seasonal variation and regions. In total, 4047 samples were collected from all seven regions in China representing distinct geographical areas: Northern, Northeastern, Eastern, Central, Southern, Southwestern, and Northwestern Regions. Samples were collected from catering, retail, and wholesale sources in different seasons throughout the course of the year. The presence of Listeria monocytogenes, Salmonella spp. , Staphylococcus aureus and diarrheagenic Escherichia coli in all the samples was examined by China's national food safety standard method. All the surveillance data were analyzed according to meat product category, market distribution, packaged form, seasonal variation and regions, respectively. Microbial prevalence of L. monocytogenes, Salmonella spp., S . aureus and diarrheagenic E. coli were detected in 1.43% (57/3974), 0.64% (26/4035), 0.79% (32/4047) and 1.06% (40/3774), respectively. The prevalence of heat-treated category RTE meat products was high, probably due to cross contamination. Prevalence data for Salmonella spp. and S. aureus were higher in the third quarter of the year. We found packaging method played a consistent role in the prevalence of all four food-borne pathogens in packaged ready-to-eat meat products. Bulk ready-to-eat meat products were significantly more likely to contain contamination than portion packaged products. In all, this study implied a potential public health problem. It is necessary to improve control measures of RTE meat products. [ABSTRACT FROM AUTHOR]

Background and Objectives: Diarrheagenic Escherichia coli (DEC) strains are a major cause of intestinal syndromes in the developing countries. The aim of this study was to determine the prevalence of enterotoxigenic E. coli (ETEC) and enteroinvasive E. coli (EIEC) in relation to phylogenetic background from patients with diarrhea. Materials and Methods: A total of 110 E. coli isolates were obtained from diarrhea patients in Sirjan, southeast of Iran. The E. coli isolates were confirmed using biochemical and bacteriological tests. DNA of E. coli isolates was extracted by boiling method and checked for existence of ETEC (LT and ST genes) and EIEC (ipaH gene) pathotypes and also characterize the phylogenetic groups on the basis of presence or absence of the chuA, yjaA genes and an anonymous DNA fragment, TspE4. C2 by multiplex PCR. Results: Out of 110 E. coli isolates, 32 (29.09%) were positive for ETEC (LT and ST genes) and 6 (5.45%) possessed EIEC (ipaH gene) pathotypes. Isolates fall into four phylogenetic groups: A (39.09%), B1 (20%), B2 (15.45%) and D (25.45%). Phylotyping of isolates of DEC indicated they were distributed in four phylogenetic groups including A (12 isolates), B1 (7), B2 (9) and D (10). Conclusion: In this study, the DEC isolates were segregated into different phylogenetic groups. The majority of isolates belonged to phylo-groups A and D. [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the prevalence of Diarrheagenic Escherichia coli ( DEC) pathotypes, a leading cause of diarrhea worldwide, among diarrheal and healthy children, up to 5 years of age, living in the city of Botucatu, São Paulo, Brazil. DEC, investigated by PCR detection of virulence factor-encoding genes associated with the distinct pathotypes, was isolated from 18.0% of the patients, and 19.0% of the controls, with enteroaggregative E. coli ( EAEC), the most frequent pathotype, being detected in equal proportion between patients and controls (10.0%). Among the enteropathogenic E. coli ( EPEC) isolates, only one isolate was able to produce the localized adherence pattern to He La cells, being thus the only typical EPEC identified. All the remaining EPEC were classified as atypical (a EPEC), and detected in 8.0% and 8.5% of the patients and controls, respectively. Regarding the serotypes, 26.5% of the analyzed EPEC isolates belonged to classical EPEC-serogroups, and the only two STEC found were serotyped as O26:H11 (patient) and O119:H7 (control). Antimicrobial susceptibility tests revealed that 43.6%, 29.5% and 2.6% of the DEC isolates were resistant to ampicillin, cotrimoxazole and gentamicin, respectively. Our data indicate that EAEC remains prevalent among children living in Botucatu, and revealed atypical EPEC as emerging putative diarrheal agents in this geographical region. [ABSTRACT FROM AUTHOR]

Introduction: Diarrheagenic Escherichia coli (DEC) represents one of the leading cause of diarrhoea in developing countries. In this study a molecular approach was applied for the detection of diarrheagenic Escherichia coli (DEC) circulating in Maputo, Mozambique. Methodology: All isolates were PCR tested for species-specific genes and for 11 molecular markers: stx1, stx2, eae, bfpA, lt, st, ipaH, aap, aggR CVD432 and daaE. Results: Of the 80 E. coli isolated, 74% were potential DEC: 21% EIEC, 19% EPEC, 15% EAEC, 13% ETEC, 5% DAEC and 1% hybrids. Conclusion: This study revealed the complexity of the etiology of diarrhea caused by pathogenic E. coli in Mozambique, and the risk of the emergence of new pathogenic variants due to the horizontal transmission of pathogenicity factors. [ABSTRACT FROM AUTHOR]

Escherichia coli colonizes the human intestine shortly after birth, with most strains engaging in a commensal relationship. However, some E. coli strains have evolved toward acquiring genetic traits associated with virulence. Currently, five categories of enteroadherent E. coli strains are well-recognized, and are classified in regard to expressed adhesins and the strategy used during the colonization. The high morbidity associated with diarrhea has motivated investigations focusing on E. coli adhesins, as well on factors that inhibit bacterial adherence. Breastfeeding has proved to be the most effective strategy for preventing diarrhea in children. Aside from the immunoglobulin content, glycocompounds and oligosaccharides in breast milk play a critical role in the innate immunity against diarrheagenic E. coli strains. This review summarizes the colonization factors and virulence strategies exploited by diarrheagenic E. coli strains, addressing the inhibitory effects that oligosaccharides and glycocompounds, such as lactoferrin and free secretory components, exert on the adherence and virulence of these strains. This review thus provides an overview of experimental data indicating that human milk glycocompounds are responsible for the universal protective effect of breastfeeding against diarrheagenic E. coli pathotypes. [ABSTRACT FROM AUTHOR]

Abstract: Escherichia coli that causes diarrhea in humans is referred to as diarrheagenic E. coli (DEC), and has been categorized into the following 5 groups: shigatoxin-producing E. coli (STEC), enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC), enteroaggregative E. coli (EAggEC), and enterotoxigenic E. coli (ETEC). In this study, we developed a novel one-step multiplex polymerase chain reaction (mPCR) for the rapid detection of 10 pathogenic genes (stx1, stx2, eae, bfpA, invE, aggR, esth, estp, elt, and astA) of DEC. Five categorized strains were used as positive controls for DEC harboring each pathogenic gene, and 828 DEC-like strains, isolated from diarrheal stool samples and assumed to be DEC on the basis of serotyping, were used in the mPCR-based detection of the pathogenic genes. To demonstrate the utility of mPCR, the 828 strains were subjected to our optimized protocol, and the results obtained were compared with those obtained by monoplex PCR. The results showed agreement for all strains. Using mPCR, we also detected 65 DEC and 41 astA-positive E. coli, and 7 of these DEC strains were “O antigen untypable” (OUT). This novel mPCR protocol allowed for rapid, convenient, and economical pathogenicity-based identification of the DEC. [Copyright &y& Elsevier]

We analyzed a randomly selected group of 30 diffusely adherent (DAEC), 30 enteropathogenic, 30 enteroaggregative, and five Shiga toxin-producing Escherichia coli strains isolated from children with diarrhea. Enterotoxigenic E. coli (ETEC) colonization factors (CFs) were evaluated by a dot-blot assay using 21 CF-specific monoclonal antibodies. Out of 95 non-ETEC strains, three DAEC were found to express coli surface antigen 20 (CS20). No other E. coli expressed CFs. We confirmed the three CS20-positive strains as ETEC-negative by repeat PCR and as toxin-negative by ganglioside-GM1-enzyme-linked immunosorbent assay. To our knowledge, this is the first study that has identified currently recognized CFs in non-ETEC diarrheagenic E. coli strains identified using molecular methods. CFs may be an unrecognized relevant adherence factor in other E. coli, which may then play a role in pathogenesis and the immune response of the host. [ABSTRACT FROM AUTHOR]

Atypical enteropathogenic Escherichia coli (aEPEC) has been associated with infantile diarrhea in many countries. The clonal structure of aEPEC is the object of active investigation but few works have dealt with its genetic relationship with other diarrheagenic E. coli (DEC). This study aimed to evaluate the genetic relationship of aEPEC with other DEC pathotypes. The phylogenetic relationships of DEC strains were evaluated by multilocus sequence typing. Genetic diversity was assessed by pulsed-field gel electrophoresis (PFGE). The phylogram showed that aEPEC strains were distributed in four major phylogenetic groups (A, B1, B2 and D). Cluster I (group B1) contains the majority of the strains and other pathotypes [enteroaggregative, enterotoxigenic and enterohemorrhagic E. coli (EHEC)]; cluster II (group A) also contains enteroaggregative and diffusely adherent E. coli; cluster III (group B2) has atypical and typical EPEC possessing H6 or H34 antigen; and cluster IV (group D) contains aEPEC O55:H7 strains and EHEC O157:H7 strains. PFGE analysis confirmed that these strains encompass a great genetic diversity. These results indicate that aEPEC clonal groups have a particular genomic background – especially the strains of phylogenetic group B1 – that probably made possible the acquisition and expression of virulence factors derived from non-EPEC pathotypes. [ABSTRACT FROM AUTHOR]

Abstract: To evaluate traditional colony-based examinations of diarrheagenic Escherichia coli (DEC), we analyzed the proportions of 5 categories of DEC among E. coli in stool specimens from patients with gastroenteritis using real-time polymerase chain reaction with novel primers and probes. Among 81 DEC isolates, 48 (59.3%) were present at proportions of ≤10%, whereas only 17 (21.0%) reached >50%. Low concentrations (≤10%) of DEC were found, particularly in most (71.8%) stool specimens collected within 48 h after the onset of illness, although such specimens were conventionally collected as close to the time of diarrhea onset as possible. Because the probability of detecting ≤10% DEC by colony-based examinations is very low, traditional laboratory methods might not detect most DEC infections, especially at the start of gastroenteritis. Thus, a diagnosis of DEC infections requires a molecular method that targets not individual colonies but E. coli clusters. [Copyright &y& Elsevier]

Diarrheagenic Escherichia coli (DEC) is a set of the most common pathogens causing diarrhea. DEC strains are classified into five pathotypes based on the possession of different virulence genes: enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC) or Shiga toxin-producing E. coli (STEC), enteroaggregative E. coli (EAEC), enterotoxigenic E. coli (ETEC), and enteroinvasive E. coli (EIEC). The development of an easy-to-use method to detect the specific virulence genes and distinguish the pathotypes is essential for the diagnosis and surveillance of DEC infections. In this study, a multiplex PCR assay (mPCR) specific to nine virulence genes and an internal control was designed for the identification of five DEC pathotypes. A temperature switch PCR (TSP) strategy was used in the PCR amplification. The PCR products were detected by capillary electrophoresis. The limit of detection (LOD) of the 10-plex reaction was 5 × 103 copies/reaction for stx2 and 5 × 102 copies/reaction for the other targets. The mPCR showed very high specificity, and inclusivity and exclusivity were both 100%. When the mPCR assay was used for the detection of 221 cryopreserved diarrhea specimens, DEC colonies were detected from 49 specimens, and the positive rate was 22.2%. The mPCR assay was sensitive and specific, and the amplified product could be analyzed easily. Thus, this method could be used effectively to identify the suspected colonies of DEC in the primary culture of the specimen. • Nine virulence genes of five DEC pathotypes can be detected in one reaction. • Capillary electrophoresis analysis makes the reading of the results easier. • The assay is sensitive and specific and could identify the suspected colonies of DEC. [ABSTRACT FROM AUTHOR]

Objectives: This study aimed to estimate diarrheagenic Escherichia coli (DEC) prevalence among pediatric patients with diarrhea at the Costa Rican National Children's Hospital-Social Security Service (Hospital Nacional de Niños-Caja Costarricense del Seguro Social; HNN-CCSS). DEC variations with respect to rainfall, presence of coinfections, and DEC antimicrobial resistance were also investigated. Results: A retrospective observational study from January 2008 to December 2016 was conducted. A total of 12 247 gastroenteritis records were analyzed. Annual DEC prevalence ranged from 2.7% (2008) to 9.0% (2013). The most prevalent pathotypes were enteroaggregative E. coli (EAEC) [n = 189 (31%)], enteropathogenic E. coli (EPEC) [n = 145 (24%)] and enteroinvasive E. coli (EIEC) [n = 91 (15%)]. A reduction in the probability of EAEC gastroenteritis was detected as rainfall rose above 200 mm/mo. [(Generalized Additive Model (GAM), p = 0.04)]. Coinfections were observed mainly between EPEC and Campylobacter spp. (10%). Antimicrobial resistance occurred in 0.6%, 29%, and 42% of DEC for ciprofloxacin, trimethoprim/sulfamethoxazole, and ampicillin, respectively. [ABSTRACT FROM AUTHOR]