Search

Foodborne diseases (FBDs) have a large disease burden among children. The major type of FBD in children is diarrhea, caused mainly by contaminated food. One of the diarrhea pathogens is Diarrheagenic Escherichia coli (DEC). The aim of this study was to establish a model of microbial prediction (DEC) in stool, caused by the transmission of FBDs in elementary schoolchildren. An observational analytic study was conducted, with a nested case-control study design. In Stage I, the study population was children in a selected elementary school at Surabaya. The sample size for Stage I was 218 children. In Stage II, the case sample was all children with a positive test for DEC (15 children), and the control sample was all children who had tested negative for DEC (60 children). The result of the laboratory tests showed that the proportion of DEC in children was 6.88% (15 of 218 children) and the proportion of Escherichia coli O157:H7 in children was only 0.46%. The most significant mode of transmission included in the model was the snacking frequency at school and the risk classification of food that was often purchased at school. The formulation of the predicting model of DEC in stool can be used as an early warning against the incidence of FBDs in elementary schoolchildren.

Background: Diarrheagenic Escherichia coli (DEC) strains are a main cause of diarrhea worldwide in children under 5 years old. DEC virulence is strongly regulated by environmental conditions and metabolites produced by the gut microbiota in the intestinal tract. In this study, we evaluated changes in gut microbiota-metabolome in children with or without diarrhea produced by DEC pathotypes. Goal: To determine gut microbiota composition and metabolome in stool samples obtained from healthy children and children with diarrhea positive for DEC pathotypes. Methods: We analyzed a total of 16 age-paired stool samples: 8 diarrheal samples positive for one DEC pathotype and 8 stool samples from healthy children. To identify the microbiota composition, we sequenced the V3-V4 region of the 16S rRNA and determined operational phylogenetic units (OPU). OPU were then used to predict metabolic pathways using the PICRUSt2 software. The presence of metabolites in stool samples was determined by LC-MS. A correlation analysis was performed with the main genera from each group and main metabolites. Bacteria associated with variance of main metabolites were identified using the MIMOSA2 software. Results: DEC and healthy groups showed a statistically different microbiota composition. A decrease in Firmicutes together with an increase in Bacteroidetes and Proteobacteria was found in the DEC group compared to the healthy group. Metabolic pathway predictions based on microbiota diversity showed that pathways involved in histidine and L-ornithine metabolism were significantly different between groups. A total of 88 metabolites detected by LC-MS were included in the metabolome analysis. We found higher levels of histamine and lower levels of ornithine in DEC samples than in the healthy group. Histamine and L-ornithine were associated with a specific microbiota species and the corresponding metabolic pathways. Conclusion: Stool samples from healthy children and children positive for DEC displayed a differential metabolome and microbiota composition. A strong correlation between a gut microbiota species and certain metabolites, such as histamine and L-ornithine, was found in the DEC group. This information might be useful to identify mechanisms and signaling molecules involved in the crosstalk between microbiota and DEC pathotypes.

The Journal of Water and Health issues a formal retraction relation to the above article by Ehssan H. Moglad et al. This decision has been taken as not all authors agree to the publication of the manuscript.

Diarrheagenic Escherichia coli as an enteropathogen has caused serious public safety problems, especially in children. A fast, easy-to-use, high-throughput method is urgently needed for diarrheagenic Escherichia coli strain identification and monitoring. In this study, we developed a portable microfluidic device based on circular fluorescent probe-mediated isothermal nucleic acid amplification for accurate and rapid detection of twelve virulence factor genes of five diarrheagenic Escherichia coli strains for point-of-care testing. This microfluidic system showed excellent performance in identifying five diarrheagenic Escherichia coli strains within 60 min, when applied in 67 clinical samples collected from hospitals for identifying diarrheagenic Escherichia coli, with a good clinical sensitivity (96.9%), specificity (97.1%) and stability (CV < 5.0%). This integrated microfluidic system is a promising diagnostic tool for applications in the identification and monitoring of diarrheagenic Escherichia coli epidemics worldwide, particularly in developing countries.

Studies evaluating the occurrence of enteropathogenic bacteria in urban rats (Rattus spp.) are scarce worldwide, specifically in the urban environments of tropical countries. This study aims to estimate the prevalence of diarrhoeagenic Escherichia coli (DEC) and Salmonella spp. with zoonotic potential in urban slum environments. We trapped rats between April and June 2018 in Salvador, Brazil. We collected rectal swabs from Rattus spp., and cultured for E. coli and Salmonella spp., and screened E. coli isolates by polymerase chain reaction to identify pathotypes. E. coli were found in 70% of Rattus norvegicus and were found in four Rattus rattus. DEC were isolated in 31.3% of the 67 brown rats (R. norvegicus). The pathotypes detected more frequently were shiga toxin E. coli in 11.9%, followed by atypical enteropathogenic E. coli in 10.4% and enteroinvasive E. coli in 4.5%. From the five black rats (R. rattus), two presented DEC. Salmonella enterica was found in only one (1.4%) of 67 R. norvegicus. Our findings indicate that both R. norvegicus and R. rattus are host of DEC and, at lower prevalence, S. enterica, highlighting the importance of rodents as potential sources of pathogenic agents for humans.

The inappropriate use of antibiotics in the poultry industry for prophylaxis and growth promotion has contributed to the development and spread of antibiotic resistance in zoonotic pathogens. This study aimed to understand the prevalence of pathogenic Escherichia coli (E. coli) in chicken faeces and their antibiotic susceptibility profiles. Forty-five E. coli isolated from the chicken faeces were further tested for virulence genes associated with diarrheagenic E. coli and extended-spectrum β-lactamase (ESBL) genes blaCTX, blaTEM, blaSHV and Metallo-beta-lactamase gene blaNDM. Results showed that eaeA and astA genes were detected in 60% of the isolates, followed by bfpA in 57.8 %, lt in 26.7% and st gene in 2.2% of the isolates. Further, 15.5% of the isolates produced ESBL phenotypically whereas genotypically, one isolate was positive for blaCTX, one for blaTEM, and one for the blaSHV gene. Twelve isolates (26.7%) harboured gene blaNDM. High levels of resistance were observed against cefepime (93.3%), ertapenem (78.9%), meropenem (73.6%), tetracycline (88.9%), and cotrimoxazole (75.6%). The study highlights the importance of poultry as a source of human pathogenic multidrug-resistant (MDR) E. coli and their environmental dissemination. [ J Adv Biotechnol Exp Ther 2022; 5(1.000): 148-162]

Introduction: Although diarrheagenic Escherichia coli (DEC) strains are important bacterial causative agents of diarrhoea, they are not routinely sought as stool pathogens in clinical laboratories as conventional microbiological testing are unable to distinguish between normal flora and pathogenic strains of E. coli. This study was undertaken to determine the prevalence of DEC pathotypes amongst children with and without diarrhoea and to detect specific virulent genes present in different DEC pathotypes, using real-time multiplex polymerase chain reaction (PCR) with high-resolution melting (HRM) technology. Materials and Methods: Stool samples were obtained from cases and controls. Using a set of conventional biochemical tests, E. coli strains were identified. Further, these isolates were subjected to multiplex PCR system for the detection of virulence genes of different pathotypes of DEC. Real-time multiplex PCR was performed for the detection of specific virulent genes of DEC pathotypes, using Rotor-Gene Q instrument (Qiagen) having High-resolution Melt analyser using Type-it HRM PCR kit (Qiagen) containing EvaGreen fluorescent intercalating dye. Results: In this study, we had successfully standardised two multiplex PCR assays which were found to be effective for direct detection of enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enterotoxigenic E. coli (ETEC) and enteroinvasive E. coli (EIEC). A total of 42 DEC strains were detected at an overall rate of 19.3% (n = 42), from the total 217 E. coli isolates recovered from the cases (n = 39, 17.9%) and control (n = 3, 3.8%) groups. Amongst the 42 DEC pathotypes (39 from cases and 3 from controls), EPEC (10%), EAEC (8.82%), ETEC (2.94%) and EIEC (1.18%) were found in children with diarrhoea (cases) and in children without diarrhoea (control) only EAEC (2.13%) and EPEC (4.26%) were detected. Age distribution, gender variation, seasonal variation and clinical features were also analysed Conclusion: This study helped evaluate the prevalence of DEC amongst children (

BACKGROUND: Diarrheagenic Escherichia coli (DEC) are an important cause of acute gastroenteritis in children; however, there is limited information available on the epidemiology, phylogenetics, serotyping, and antibiotic susceptibility of DEC in children in the United States. The aim of this study was to determine the molecular epidemiology of DEC among children with and without acute gastroenteritis in Davidson County, Tennessee. METHODS: This prospective, frequency matched, case-control study recruited subjects 15 days to 17 years of age and detected DEC with polymerase chain reaction from stool samples. Additional testing was done to define phylogenetics and antibiotics resistance. RESULTS: Among 1267 participants, 857 cases and 410 controls, 5.5 % were positive for at least one subtype of DEC. Enteroaggregative E. coli (EAEC) [n=32 (45%)] was the most common subtype followed by enteropathogenic E. coli (EPEC) [n=30 (43%)], Shiga toxin-producing E. coli [n=4 (6%)], and diffusely adherent E. coli [n=4 (6%)]. No significant difference in prevalence of DEC was found between cases (5%) and controls (7%) [Odds ratio (OR): 0.66 (95 % confidence interval (CI) 0.4–1.07)] and results were similar when data were stratified by subtypes and adjusted for age, sex, race, and ethnicity. Substantial diversity was found among DEC isolates in terms of phylotypes and serotypes and a large proportion was resistant to, at least, one antibiotic. CONCLUSIONS: EAEC and EPEC were frequently found in both cases and controls in this study population. DNA-based methods for detection of these subtypes need further investigation to help differentiate between pathogenic and colonizing strains.

Background and Objectives: Diarrheagenic Escherichia coli (DEC) pathotypes are important causes of diarrhea among children in developing countries. The objective of this study was to determine the frequency and antibiotic resistance pattern of DEC pathotypes in children aged less than 10 years. Methods: This cross-sectional study was done on 300 E. coli strains isolated from diarrheic stool samples of children aged less than 10 years who were admitted to hospitals and central laboratory in Zahedan, between July and October 2016. DEC pathotypes were identified by standard biochemical testing and phenotypic testing using polyvalent antiserums. Antibiotic resistant pattern of these strains was evaluated against 11 different antibiotics by the agar disk diffusion method according to the Clinical and Laboratory Standards Institute guidelines. Results: Of the 300 E. coli isolates, 89 (29.6%) were found positive for DEC using polyvalent antiserums. In this study, 35 cases (39.3%) reacted with antiserum 1, 21 cases (25.8%) reacted with the antiserum 2, and 31 cases (34.8%) reacted with antiserum 3. The highest rate of resistance was observed against ampicillin (94.8%), tetracycline (87.2%), and co-trimoxazole (70.5%). In addition, the lowest rate of resistance was related to imipenem (1%) and ciprofloxacin (8.9%). Conclusion: DEC pathotypes are the important causes of diarrhea among children admitted to hospitals of Zahedan. Considering the high rate of antibiotic resistance among these pathotypes in this region, prescription of antibiotics should be based on accurate detection of these strains. Keywords: Escherichia coli, Child, Antibiotic Resistance.

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.

Introduction: Diarrheagenic Escherichia coli (DEC) is a relevant cause of diarrhea, particularly among infants and young children in developing countries. Methodology: We compared the frequency, antimicrobial resistance, adherence, enterovirulence and genetic diversity of DEC isolates from Guarani indigenous population under five living in distinct villages in Brazil. Results: Of the 314 E. coli isolates from 57 children, with and without diarrhea, 15% (48/314) were classified in DEC categories: aEPEC (56%, 27/48), EAEC (35%, 17/48) and ETEC (8%, 4/48). ETEC belonged to plylogroup A, EAEC to groups A, B1, B2 and D, and aEPEC to phylogroups A, B1, and B2. EAEC exhibited the aggregative adherence phenotype while ETEC and aEPEC the aggregative and undefined patterns. Multidrug-resistance was detected in aEPEC, ETEC and EAEC while extensive drug-resistance was found in EAEC and aEPEC. RAPD typing revealed a genetically diverse bacterial population. Conclusion: This is the first report regarding aspects of DEC in an indigenous Brazilian population, showing that Guarani children are DEC carriers and that antimicrobial resistance at high levels is widely disseminated among these enteropathogens.

Introduction This study was aimed to evaluate the prevalence of diarrheagenic Escherichia coli (DEC) and ESBL genes in the children with gastroenteritis and to explore the predominant pathotypes of DEC with the multiplex PCR (MPCR) type. Methods To investigate the prevalence of DEC in the young children, 303 samples were obtained from children with gastroenteritis and evaluated by the MPCR test to characterize the five pathotypes of DEC. Results DEC infections were found in 68 cases (22.44%). Enteroaggregative E. coli (EAEC) was the most prevalent pathotype (80.9%, 55/68) determined by three primers sets. Detection rate for Enteropathogenic E. coli (EPEC), Enterotoxigenic E. coli (ETEC), Enteroinvasive E. coli (EIEC) and Shiga toxin producing E. coli (STEC) was 8.8% (6/68), 8.8% (6/68), 4.4% (3/68) and 4.4% (3/68), respectively. The frequency DEC was not statistically significant with age and the gender of the patients (P ˃0.05). The blaSHV gene was not found in the isolates and the blaTEM and blaCTX-M genes were identified in 29.4% and 25% of the isolates, respectively. Conclusions DEC could be regarded as one of the major causative agents of children diarrhea, thus emphasizing the role of EAEC as the main causative pathotype of DECs. Detection of a high number of ESBL genes might be associated to the limited management of antimicrobial resistance among these isolates.

Enterotoxigenic Escherichia coli (ETEC) and Shigella ssp. infections are associated with high rates of mortality, especially in infants in developing countries. Due to increasing levels of global antibiotic resistance exhibited by many pathogenic organisms, alternative strategies to combat such infections are urgently required. In this study, we evaluated the stability of five coliphages (four Myoviridae and one Siphoviridae phage) over a range of pH conditions and in simulated gastric conditions. The Myoviridae phages were stable across the range of pH 2 to 7, while the Siphoviridae phage, JK16, exhibited higher sensitivity to low pH. A composite mixture of these five phages was tested in vivo in a Galleria mellonella model. The obtained data clearly shows potential in treating E. coli infections prophylactically.

The emergence and persistence of multidrug-resistant (MDR) diarrheagenic Escherichia coli (DEC) causing acute diarrhea is a major public health challenge in developing countries. The aim of this study was to evaluate the resistance phenotypes of DEC isolated from stool samples collected from children less than 5 years of age with acute diarrhea living in Ouagadougou/Burkina Faso. From August 2013 to October 2015, this study was carried out on 31 DEC strains of our study conducted in “Centre Médical avec Antenne Chirurgicale (CMA)” Paul VI and CMA of Schiphra. DEC were isolated and identified by standard microbiological methods and polymerase chain reaction (PCR) method was used to further characterize them. Antimicrobial susceptibility testing was done based on the disk diffusion method. DEC isolates were high resistant to tetracycline (83.9%), amoxicillin (77.4%), amoxicillin clavulanic acid (77.4%), piperacillin (64.5%), and colistin sulfate (61.3%). The most resistant phenotype represented was the extended spectrum β-lactamase (ESBL) phenotype (67.7%). Aminoglycosides were 100% active on enteroinvasive E. coli (EIEC) and enterohemorrhagic E. coli (EHEC). All the DEC isolates exhibited absolute (100%) sensitivity to ciprofloxacin. Monitoring and studying the resistance profile of DEC to antibiotics are necessary to guide probabilistic antibiotic therapy, especially in pediatric patients.

Diarrheagenic Escherichia coli (DEC) is important bacteria of children's endemic and epidemic diarrhea worldwide. The aim of this study was to determine the prevalence of DEC isolated from stool samples collected from children with acute diarrhea living in Ouagadougou, Burkina Faso. From August 2013 to October 2015, stool samples were collected from 315 children under 5 years of age suffering from diarrhea in the "Centre Medical avec Antenne Chirurgicale (CMA)" Paul VI and the CMA of Schiphra. E. coli were isolated and identified by standard microbiological methods, and the 16-plex PCR method was used to further characterize them. Four hundred and nineteen (419) E. coli strains were characterized, of which 31 (7.4%) DEC pathotypes were identified and classified in five E. coli pathotypes: 15 enteroaggregative E. coli (EAEC) (48.4%), 8 enteropathogenic E. coli (EPEC) (25.8%) with 4 typical EPEC and 4 atypical EPEC, 4 enteroinvasive E. coli (EIEC) (12.9%), 3 enterohemorrhagic E. coli (EHEC) 9.67%, and 1 enterotoxigenic E. coli (ETEC) 3.2%. The use of multiplex PCR as a routine in clinical laboratory for the detection of DEC would be a useful mean for a rapid management of an acute diarrhea in children.

The source and routes of diarrheagenic Escherichia coli (DEC) remain poorly understood. To investigate the involvement of domestic animals in the dissemination of DEC, the prevalence of DEC in foods and fecal specimens from cattle, pigs, chickens, healthy carriers, and patients in Osaka and Hyogo, Japan was investigated using a multiplex real-time Polymerase Chain Reaction assay. The most abundant virulence genes were astA and eae, which had a prevalence 46.8% and 27.4%, respectively. Additionally, stx1 (26.6%) and stx2 (45.9%) were prevalent in cattle feces, while est (8.5%) and elt (7.6%) were prevalent in pig feces. afaB was the second-most prevalent gene in patients and healthy carriers, and it had detection rates of 5.1% and 8.1%, respectively. In contrast, afaB was not detected in animal feces or foods, except for three porcine fecal samples. The aggR gene was more prevalent in humans than in foods or animal feces. Both Shiga toxin-producing E. coli and atypical enteropathogenic E. coli carried by cattle may be sources for diarrheal diseases in humans. Pigs may be a source for human enterotoxigenic E. coli infections, whereas humans are expected to be the reservoir for diffusely adhering E. coli, enteroaggregative E. coli, and enteroinvasive E. coli.

Diarrheagenic E. coli (DEC) are one of the most common causes for diarrhea worldwide, especially in children. We evaluated the rapid RIDA ® GENE (RG) real-time multiplex PCR assays (R-Biopharm, Darmstadt, Germany) for the detection of the most important diarrheagenic E. coli. Three hundred fifteen liquid or non-formed stool specimens were examined. The results of the RG multiplex assays were compared to specific PCR methods. The sensitivity and specificity of the RG PCRs were as follows, 100%/100% for the detection of EHEC, 96.3% and 99% for EPEC, 100% and 100% for the detection of EAEC, ETEC and EIEC, respectively. Overall, the RG real-time PCR system for the detection of DEC tested in this study provided reliable and rapid results and shows the ability as a useful addendum for the detection of diarrheagenic E. coli in the medical laboratory.

Diarrheagenic Escherichia coli (DEC) is an important agent of endemic and epidemic diarrhea worldwide, particularly in developing countries. DEC cannot be differentiated from commensal E. coli on selective media, although there are a few exceptions. Most studies use the colony isolation method, which cannot detect low numbers of DEC, and therefore, these studies might underestimate the incidence of DEC. In the present study, we employed a colony sweep method with real-time PCR targeting virulence genes of 5 categories of DEC; this technique can detect very low numbers of DEC among hundreds of commensal E. coli. DEC was detected in 171 (55.9%) of 306 children with diarrhea in Kenya. The prevalence of DEC in Kenya was notably higher than that (30 in 143, 21.0%) in Indonesia. Occurrences of multiple DEC infection in Kenya were frequent (69 in 306, 23.2%), suggesting that the source of DEC infection may be related to grossly contaminated food and water. In contrast, only 9 (6.0%) of 150 healthy adults in Kenya carried DEC. Considering that healthy adults naturally harbor non-DEC, it is interesting how children exclude DEC but not non-DEC as they grow up. Several mechanisms, such as mucosal immunity and intestinal microbiota, might be involved in the exclusion of DEC.

Diarrheagenic Escherichia coli (DEC) is one of the leading causes of gastrointestinal disorders worldwide and an important public health challenge. DEC infection is often underdiagnosed during routine microbiological analysis, especially in resource constrained settings; the use of molecular tests could however help to determine the distribution of DEC and its clinical significance. Here, a study to assess the prevalence of DEC in clinical samples from patients

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.

A multiplex PCR assay was evaluated for diagnosis of diarrheagenic Escherichia coli in stool samples of patients with diarrhoea submitted to a diagnostic microbiology laboratory. Two procedures of DNA template preparationproteinase K buffer method and the boiling method were evaluated to examine isolates of E. coli from 150 selected diarrhoeal cases. By proteinase K buffer method, 119 strains (79.3%) of E. coli were characterized to various categories by their genes that included 55.5% enteroaggregative E. coli (EAEC), 18.5% enterotoxigenic E. coli (ETEC), 1.7% enteropathogenic E. coli (EPEC), and 0.8% Shiga toxin-producing E. coli (STEC). Although boiling method was less time consuming (

Diarrheagenic Escherichia coli (DEC) are the most common agents of diarrhea. Waterborne DEC could pose a potential health risk to human through agricultural, household, recreational, and industrial use. There are few published reports on the detection of DEC and its seasonal distribution in aquatic environments. The presence of DEC in different types of aquatic environments was investigated in this study. Water samples were collected from major rivers, water reservoirs, and recreational hot springs throughout Taiwan. Moreover, an intensive water sampling plan was carried out along Puzih River. The detection of DEC target genes was used to determine the presence of enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), and Shiga toxin-producing E. coli (STEC). Among the 383 water samples analyzed, DEC was found in 122 (31.8%) samples. The detection rate varied by genotype, raging from 3.6% for STEC to 17.2% for EPEC. The DEC detection rate was higher from river waters than reservoirs and hot springs. In addition, DEC was detected at a higher rate in spring and summer. The presence of EPEC was significantly associated with total coliform levels among hot spring samples. Moreover, the presence of ETEC in river water samples was associated with heterotrophic plate counts. Water with EPEC differed significantly in pH from Puzih River samples. These results suggest that seasonal characteristics may affect the presence of DEC in different aquatic environments, and water quality indicators may be indicative of the presence of DEC.

Purpose: Seasonal rains in Pakistan result in heavy floods across the country, whereby faecal contaminants will be added to the water bodies and cause numerous food-borne outbreaks. The present study was aimed to determine the prevalence of diarrheagenic Escherichia coli (DEC) strains in the water sources. Materials and Methods: Two hundred water samples collected during (2011–2012) were processed for the isolation of E. coli (EC) strains. EC strains were further analysed for antibiotic susceptibility patterns, and pathogroups-specific virulence factors stx1, stx2, stx2c, eae, tir, hlyA, bfpA, estA and eltA were detected using multiplex polymerase chain reaction. Results: Thirty-three percent of the water samples were contaminated with EC pathotypes. Fifty percent (33/66) of the DEC pathotypes were identified as enterotoxigenic EC (ETEC). Seventy-two percent (13/18) of the enteropathogenic EC (EPEC) strains were identified as typical EPEC and 28% (5/18) as atypical EPEC. Eleven percent (7/66) of the Shiga toxin EC (STEC) isolates carried a combination of stx1 and stx2 genes. Summer was found as a peak season with 47% (31/66) for EC pathogroups' activities. Eighty-nine percent of the strains showed resistance against tetracycline. Conclusion: ETEC and EPEC are the primary causes of water contamination in southern regions of Khyber Pakhtunkhwa province, Pakistan. Firm adherence to the prescribed drugs can decrease trends in antibiotic resistance.

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.