Diffusely Adherent E. Coli Research Articles

Aksaray İli Buzağı İshallerinde Escherichia coli ve Escherichia coli Bakterisinin Son Üç Aylık Antibiyogram Sonuçları

Escherichia coli is a gram-negative, facultative anaerobic, motile, non-spore-forming rod-shaped bacterium belonging to the Enterobacteriaceae family. Pathogenic E. coli are divided into two groups: extraintestinal and intestinal. Intestinal Escherichia coli pathotypes: enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), Vero- or Shiga-toxin-producing E. coli (VTEC or STEC), enterohemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAEC), enteroinvasive E. coli (EIEC) and diffusely adherent E. coli (DAEC). Extraintestinal pathogenic E. coli: These can be listed as septicemic pathogenic E. coli, uropathogenic E. coli, avian pathogenic E. coli, breast pathogenic E. coli and those that cause uterine infections, endometrial pathogenic E. coli. Enterotoxigenic Escherichia coli (ETEC) is the most common cause of neonatal diarrhea in calves and is a bacterial disease that colonizes the small intestine, produces enterotoxin, and occurs among calves during the neonatal period. The aim of this study is to determine the E.coli isolates that cause neonatal calf diarrhea in the Aksaray region between January, February and March 2021. To be used in the study, internal organ samples (liver, heart, lung and mesenteric lymph node) of neonatal calves that died due to diarrhea were taken from 20 different cattle farms in the Aksaray region. One calf that died in each farm was included in the sampling, and a total of 20 calves were sampled. E.coli was isolated in 12 (60%) of the internal organ samples covering these three months. According to the antibiogram results of the samples, 50% of the isolates were converted to Amoxicillin and Erythromycin, 33.3% to Tetracycline, 58.3% to Trimethoprim-sulfamethoxazole, 66.6% to Streptomycin, 75% to Flofenicol, Gentamicin and Enrofloxacin, % 83.3 of them were found to be sensitive to Cefloxacin and Cefloxacinium.

Detection and antibiotic resistance of diarrheagenic Escherichia coli from patients with diarrhea in Ulaanbaatar, Mongolia.

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. 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). 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. 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.

Phylogrouping and characterization of Escherichia coli isolated from colonic biopsies and fecal samples of patients with flare of inflammatory bowel disease in Iran.

BackgroundAlthough the etiopathogenesis of inflammatory bowel disease (IBD) is still poorly understood, Escherichia coli has been described as a potential causative microorganism in IBD pathogenesis and also disease progression, offering a potential therapeutic target for disease management. Therefore, we conducted this study to investigate the pathotypes, phylogenetic groups, and antimicrobial resistance of E. coli isolates from patients with IBD in Iran.MethodsFecal and biopsy colonic samples were collected from IBD patients experiencing flare-up episodes referred to Taleghani hospital in Tehran, Iran, between August 2020 and January 2021. Identification of E. coli strains was performed based on biochemical and molecular methods. Antibiotic susceptibility testing was performed as recommended by the Clinical and Laboratory Standards Institute. Phylogrouping and pathotyping of each isolate were carried out using polymerase chain reaction (PCR) and multilocus sequence typing (MLST) assays.ResultsA total of 132 non-duplicate E. coli strains were isolated from 113 IBD patients, including 96 ulcerative colitis (UC), and 17 Crohn’s disease (CD) patients. In our study, 55% of CD-related E. coli and 70.5% of UC-related isolates were non-susceptible to at least three or more unique antimicrobial classes, and were considered as multidrug-resistant (MDR) strains. E. coli strains exhibited a high level of resistance to cefazolin, ampicillin, tetracycline, ceftazidime, ciprofloxacin, and cefotaxime. Enterotoxigenic E. coli (ETEC) and diffusely adherent E. coli (DAEC) were the most prevalent pathotypes, and groups B2 and D were the predominant phylogroups.ConclusionIn the present study, we found that E. coli strains that colonize the gut of Iranian patients with IBD most frequently belonged to phylogenetic groups B2 and D. We also conclude that E. coli isolates from IBD patients have been revealed to be resistant to commonly used antibiotics, in which most of them harbored strains that would be categorized as MDR.

Bacterial diarrhea among infants in developing countries: An overview of diarrheagenic Escherichia coli (DEC)

Many cases of diarrhea are due to an infection in the gastrointestinal tract caused by microorganisms such as bacteria, viruses and parasites. This may be present alone or can be together with different symptoms, such as vomiting, abdominal pain, nausea and weight loss. There is an estimate of 2 billion cases of diarrhea disease globally that happens yearly. Also, approximately 1.9 million children globally below the age of 5 years die from diarrhea every year mostly in the developing countries making it the second leading cause of mortality in children of this age group. Recurring and regular diarrhea can affect growth of children and cognitive development and increase their vulnerability to different communicable diseases. About 89% of diarrhea related deaths are caused by insufficient hygiene, bad water, and inadequate sanitation. Major cause of diarrhea in children is Diarrheagenic Escherichia coli (DEC) which is acquired through water and food that is contaminated. There are 6 main pathotypes:, diffusely adherent E. coli (DAEC), enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), shiga toxin–producing E. coli (STEC), enteroinvasive E. coli (EIEC), and enteroaggregative E. coli (EAEC). Adherent invasive E. coli (AIEC) is is the new pathotype that has been recently reported. Biochemical tests, culture and modern methods like Polymerase Chain Reaction are the conventional techniques carried out in the identification of E. coli species. Preventive measures and treatment includes proper sanitation, improved hygeine, access to safe drinking-water, exclusive breastfeeding of the babies for the first 6 months of life, proper rehydration with oral rehydration salts (ORS) or intravenous fluids in case of shock, early hospitalization for children who are extremely malnourished, zinc supplementation, and in some circumstances, the use of antibiotics which should be implemented to reduce the burden of diarrhea among children.

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

The aim of the study was to determine the frequency of six genes encoding serine protease autotransporter proteins Enterobacteriaceae (SPATE) in diffusely adherent Escherichia coli (DAEC) isolates from children with (WD, n=63) and without diarrhea (WOD, n=41) from Lima, Peru. WOD were considered a control group. For the detection of the genes, 2 multiple PCRs were standardized: triple A (sigA, pet, espP) and triple B (sat, pic, espC). In both groups, the most frequent SPATE gene was Sat (39.7% of WD and 41.5% of WOD), followed by spP (20.6% and 9.7% in WD and WOD respectively). The other genes were detected in proportions lower than 10.0%, in the following order of frequency: pet, sigA, espC and pic, without significant differences between the groups. It was concluded that Sat is the most frequent SPATE in DAEC and that these strains may possess SPATE genes regardless of whether they are isolated in WD or WOD.

Pathotyping and Antibiotic Resistance Profiling of Escherichia coli Isolates from Children with Acute Diarrhea in Amatole District Municipality of Eastern Cape, South Africa.

Background Diarrhea has been reported as the leading cause of childhood mortality and morbidity globally but with disproportionate impacts in developing nations. Among bacterial etiologic agents of diarrhea, diarrheagenic Escherichia coli is the main cause of the disease among children under the age of 5 years. This study is aimed at determining the prevalence and antibiogram pattern of diarrheagenic Escherichia coli (DEC) pathotypes associated with diarrhea cases in the study area. Methods A total of 120 presumptive isolates of E. coli were obtained from diarrheal stool samples from male and female patients below 12 years of age using chromogenic agar. Confirmation of the isolates and screening for virulence genes were determined by polymerase chain reaction (PCR) while antimicrobial susceptibility testing was performed using the disk diffusion method. The presence of antibiotic resistance genes to chloramphenicol and tetracycline among the confirmed isolates was also profiled by PCR based on the observed phenotypic resistance pattern. Results Of the 120 presumptive isolates, 88.3% (106/120) were confirmed as E. coli through PCR. The molecular pathotyping of the confirmed isolates showed their distribution as 41% (43/106) of diffusely adhering E. coli (DAEC), 17% (18/106) of enterohemorrhagic E. coli (EHEC), 17% (18/106) of enteropathogenic E. coli (EPEC), and 10% (11/106) of enteroinvasive E. coli (EIEC), while enteroaggregative E. coli (EAEC) and enterotoxigenic E. coli (ETEC) were not detected, and the remaining 15% did not belong to any pathotype. Notably, high resistance of the isolates to commonly used antimicrobials was observed as follows: ampicillin (98%), chloramphenicol (94%), trimethoprim-sulfamethoxazole (96%), and tetracycline (90.6%), while a relatively low number of the confirmed isolates were resistant to ciprofloxacin (45%) and imipenem (36%). In addition, 94% of the isolates that exhibited phenotypic resistance against chloramphenicol harbored the catA1 resistance gene while 89% that showed resistance to tetracycline had tetA genes. Conclusions These findings showed that DEC could be considered as the leading etiologic bacterial agent responsible for diarrhea in the study community, and the observable high degree of resistance of the isolates to antimicrobial agents is of huge significance, calling for stakeholders to adopt and consolidate the existing antimicrobial stewardship scheme of the government, in order to ensure an uncompromised public health.

New Insights Into DAEC and EAEC Pathogenesis and Phylogeny.

Diarrheagenic E. coli can be separated into six distinct pathotypes, with enteroaggregative (EAEC) and diffusely-adherent E. coli (DAEC) among the least characterized. To gain additional insights into these two pathotypes we performed whole genome sequencing of ten DAEC, nine EAEC strains, isolated from Mexican children with diarrhea, and one EAEC plus one commensal E. coli strains isolated from an adult with diarrhea and a healthy child, respectively. These genome sequences were compared to 85 E. coli genomes available in public databases. The EAEC and DAEC strains segregated into multiple different clades; however, six clades were heavily or exclusively comprised of EAEC and DAEC strains, suggesting a phylogenetic relationship between these two pathotypes. EAEC strains harbored the typical virulence factors under control of the activator AggR, but also several toxins, bacteriocins, and other virulence factors. DAEC strains harbored several iron-scavenging systems, toxins, adhesins, and complement resistance or Immune system evasion factors that suggest a pathogenic paradigm for this poorly understood pathotype. Several virulence factors for both EAEC and DAEC were associated with clinical presentations, not only suggesting the importance of these factors, but also potentially indicating opportunities for intervention. Our studies provide new insights into two distinct but related diarrheagenic organisms.

Commonality of adherent-invasive Escherichia coli isolated from patients with extraintestinal infections, healthy individuals and the environment.

Adherent-invasive Escherichia coli (AIEC) has been implicated as a microbiological factor in inflammatory bowel disease (IBD) pathogenesis. These strains are defined by their ability to adhere to and invade intestinal epithelial cells, and to survive and replicate in macrophages. We postulated that AIEC strains may commonly inhabit the gut of healthy individuals (HI), cause extraintestinal infections, and be found in sewage treatment plants (STP) and surface waters (SW). A total of 808 E. coli strains isolated from HI; patients with community-acquired urinary tract infection (CA-UTI), septicaemia and urosepsis; STP; and SW, showing a diffuse adhesion pattern to Caco-2 cells were included in this study. Typing of the strains using a combination of RAPD-PCR and PhPlate fingerprinting grouped them into 48 common clones (CCs). Representatives of each CC were tested for the ability to invade Caco-2 cells, survive and replicate in macrophages, and for the presence of six virulence genes commonly found among AIEC strains. Twenty CCs were deemed AIEC based on their ability to survive and replicate in macrophages, while encoding htrA, dsbA and clbA genes. These CCs primarily originated from HI and CA-UTI patients but were also detected in secondary locations including STP and SW. Strains lacking intramacrophagic survival and replication abilities were regarded as diffusely adhering E. coli (DAEC). Certain clones of AIEC are common in the gut of HI whilst promoting CA-UTI. The survival and persistence of AIEC in STP and SW may have serious public health ramifications for individuals predisposed to IBD.

Characterization and antibiotic resistance pattern of diffusely adherent Escherichia coli (DAEC), isolated from paediatric diarrhoea in Shiraz, southern Iran.

Diarrhoea is a major health concern, especially in developing countries. Research has implicated diffusely adherent Escherichia coli (DAEC) strains as a cause of diarrhoea. In this study, we investigated the prevalence, adherence assay, virulence gene profiles and antimicrobial resistance of DAEC at a hospital in southern Iran. In this cross-sectional study, 309 infants and children under the age of 13 years with diarrhoea who had been referred to Shahid Dastgheib Hospital, Shiraz between October 2018 and May 2019 were recruited. Microbiological methods, PCR, HEp-2 adherence assay and antimicrobial susceptibility test were used. Of the 309 stool samples, 207 (66.9%) were found to contain E. coli by biochemical tests and culture. Molecular analysis of Afa/Dr and AIDA-I adhesin-encoding genes showed that 14 (6.7%) out of 207 E. coli isolates were DAEC. All DAEC isolates in HEp-2 cells showed a diffusely adherent pattern. The virulence genes sat, pet, sigA, pic, astA and fimH were found in 50%, 0%, 14.2%, 14.2%, 21.4% and 100% of DAEC isolates, respectively. The most effective antibiotic against the DAEC isolates was imipenem (92.8%) and the least effective was ampicillin (0%). Our findings expand the knowledge on DAEC prevalence and its characteristics in Iran. It also explains the role of virulence genes in DAEC pathogenesis. The results showed that although the prevalence of DAEC is low, these strains exhibit a high rate of antimicrobial resistance as well as high frequency for carrying virulence genes.

Occurrence, pathotypes, and antimicrobial resistance profiles of diarrheagenic Escherichia coli strains in animal source food products from public markets in Mashhad, Iran

Diarrheagenic Escherichia coli (DEC) strains are common pathogens that transmitted through the consumption of contaminated foods, and cause acute intestinal diseases in human. The present study was conducted on 300 samples including raw milk, meat and some of their products from November 2016 to October 2017. Microbiological and molecular diagnostic techniques were used to identification of DEC. A total of 69% (207/300) E. coli was isolated and the frequency of DEC was 25.6% (53/207). The pathogenic groups of DEC recovered from the isolates had the following profile: Shiga-like toxin producing E. coli (STEC): 62.3%, enteropathogenic E. coli (EPEC): 24.5%, enteroaggregative E. coli (EAEC): 9.4%, and enterotoxigenic E. coli (ETEC): 3.8%. Neither enteroinvasive E. coli (EIEC) nor diffusely adherent E. coli (DAEC) were found. The highest incidence values were found in hamburgers (46.7%), and the highest rate of DEC positive prevalence was in March. Of the DEC strains, 90.6% (48/53) displayed resistance to at least one antibiotic and the highest level of resistance was found for tetracycline (69%). The obtained results revealed that the studied animal source food products may easily act as a reservoir of DEC with a potential ability to transfer antibiotic resistance and virulence genes to the gastrointestinal microbiota. Therefore, it is of paramount importance to develop effective strategies for improving food safety and updated guidelines for the prudent use of antimicrobial agents in Iran.

Low frequency of enterohemorrhagic, enteroinvasive and diffusely adherent Escherichia coli in children under 5\u2009years in rural Mozambique: a case-control study

BackgroundDiarrheagenic Escherichia coli (DEC) are among the leading pathogens associated with endemic diarrhea in low income countries. Yet, few epidemiological studies have focused the contribution of enterohemorrhagic E. coli (EHEC), enteroinvasive E. coli (EIEC) and diffusely adherent E. coli (DAEC).MethodsWe assessed the contribution of EHEC, EIEC and DAEC isolated from stool samples from a case-control study conducted in children aged < 5 years in Southern Mozambique between December 2007 and November 2012. The isolates were screened by conventional PCR targeting stx1 and stx2 (EHEC), ial and ipaH (EIEC), and daaE (DAEC) genes.ResultsWe analyzed 297 samples from cases with less-severe diarrhea (LSD) matched to 297 controls, and 89 samples from cases with moderate-to-severe diarrhea (MSD) matched to 222 controls, collected between November 3, 2011 and November 2, 2012. DEC were more common among LSD cases (2.7%, [8/297] of cases vs. 1.3% [4/297] of controls; p = 0.243]) than in MSD cases (0%, [0/89] of cases vs. 0.4%, [1/222] of controls; p = 1.000). Detailed analysis revealed low frequency of EHEC, DAEC or EIEC and no association with diarrhea in all age strata. Although the low frequency, EIEC was predominant in LSD cases aged 24–59 months (4.1% for cases vs. 0% for controls), followed by DAEC in similar frequency for cases and controls in infants (1.9%) and lastly EHEC from one control. Analysis of a subset of samples from previous period (December 10, 2007 and October 31, 2011) showed high frequency of DEC in controls compared to MSD cases (16.2%, [25/154] vs. 11.9%, [14/118], p = 0.383, respectively). Among these, DAEC predominated, being detected in 7.7% of cases vs. 17.6% of controls aged 24–59 months, followed by EIEC in 7.7% of cases vs. 5.9% of controls for the same age category, although no association was observed. EHEC was detected in one sample from cases and two from controls.ConclusionsOur data suggests that although EHEC, DAEC and EIEC are less frequent in endemic diarrhea in rural Mozambique, attention should be given to their transmission dynamics (e.g. the role on sporadic or epidemic diarrhea) considering that the role of asymptomatic individuals as source of dissemination remains unknown.

The prevalence of mucosa-associated diffusely adherent Escherichia coli in children with inflammatory bowel disease.

The relationship of diffusely adherent Escherichia coli (DAEC) with pediatric inflammatory bowel disease (IBD) has not been previously studied. Diffusely adherent E. coli are a common cause of long-lasting childhood diarrhea and we postulated that they may induce inflammation of the intestinal mucosa, contributing to the development of IBD in susceptible children. The aim of the study was to investigate the relationship between DAEC and pediatric IBD, including Crohn's disease (CD) and ulcerative colitis (UC). Diffusely adherent E. coli isolates were also assessed regarding their pathogenicity. Diffusely adherent E. coli were screened among 130 E. coli strains isolated from intestinal biopsy specimens from 26 children with IBD using polymerase chain reaction (PCR) with primers specific to the pathotype and adherence assays to HEp-2 cells. Diffusely adherent E. coli were further analyzed for their ability to adhere to and invade polarized Caco-2 cells. The immunomodulatory effect of DAEC on the secretion of tumor necrosis factor α (TNF-α) by human monocyte-derived macrophages (MDM) was assessed using an immunoenzymatic assay. Diffusely adherent E. coli were recovered from 18 (69.2%) of the 26 intestinal biopsy specimens from both CD and UC patients. Most DAEC isolates carried AfaE3 adhesin, adhered to and were internalized by Caco-2 cells, and induced secretion of elevated levels of TNF-α. The study demonstrated the internalization of DAEC by intestinal epithelial cells and their ability to induce secretion of increased level of TNF-α in a Caco-2/macrophage compartmentalized culture. This indicated that the pathovar should be considered a pathobiont inducing inflammation of the intestinal mucosa in pediatric patients with IBD.

Escherichia coli Pathobionts Associated with Inflammatory Bowel Disease.

Gut bacteria play a key role in initiating and maintaining the inflammatory process in the gut tissues of inflammatory bowel disease (IBD) patients, by supplying antigens or other stimulatory factors that trigger immune cell activation. Changes in the composition of the intestinal microbiota in IBD patients compared to that in healthy controls and a reduced diversity of intestinal microbial species are linked to the pathogenesis of IBD. Adherent invasive Escherichia coli (AIEC) has been linked to Crohn's disease (CD) patients, while diffusely adherent E. coli (DAEC) has been associated with ulcerative colitis (UC). Bacteriological analysis of intestinal biopsy specimens and fecal samples from IBD patients shows an increased number of E. coli strains belonging to the B2 phylogenetic group, which are typically known as extraintestinal pathogenic E. coli (ExPEC). Results from studies of both cell cultures and animal models reveal pathogenic features of these E. coli pathobionts, which may link them to IBD pathogenesis. This suggests that IBD-associated E. coli strains play a facilitative role during IBD flares. In this review, we explain IBD-associated E. coli and its role in IBD pathogenesis.

Molecular Pathotyping of Escherichia Coli Isolates and Detection of Residual Antibiotics in Raw Cow Milk in Iran

Escherichia coli is an important species within the Enterobacteriaceae including at least six pathogenic classes of enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAEC), enteroinvasive E. coli (EIEC) and diffusely adherent E. coli (DAEC). Therefore, the aims of the current study were molecular characterization of E. coli pathotypes in raw cow milk samples in Borujerd, Iran, and detection of antibiotic residues in those samples due to the importance for the public health. Totally, 150 raw cow milk samples were collected from Borujerd, Iran, 2015. Eleven genes of bfpA, eaeA, east1, flicH7, hlyA, ial, lt, st, stx1, stx2 and O157 were detected in isolates using PCR to molecularly typing the E. coli isolates. Of 150 samples, 31 E. coli strains (totally 20.6%) were isolated, including 16 ETEC (51.6%), nine EPEC (29%) and six EHEC (19.4%). No EIEC (0%) or EAEC (0%) was isolated. In PCR, 13 eaeA (29.5%), eight lt (18.18%), eight st (18.18%), four stx1 (9.09%), three stx2 (6.81%), three hlyA (6.81%), two bfpA (4.54%), two O157 (4.54%) and one flicH7 (2.27%) encoding genes were found in E. coli isolates. Furthermore, 30 samples (20%) included antibiotic residues. In General, pathotyping of the bacterial isolates can better reveal possible correlations between these pathotypes and foodborne outbreaks.

Diffusely Adherent Escherichia coli Strains Isolated from Healthy Carriers Suppress Cytokine Secretions of Epithelial Cells Stimulated by Inflammatory Substances.

Diarrheagenicity of diffusely adherent Escherichia coli (DAEC) remains controversial. Previously, we found that motile DAEC strains isolated from diarrheal patients induced high levels of interleukin 8 (IL-8) secretion via Toll-like receptor 5 (TLR5). However, DAEC strains from healthy carriers hardly induced IL-8 secretion, irrespective of their possessing flagella. In this study, we demonstrated that SK1144, a DAEC strain from a healthy carrier, suppressed IL-8 and IL-6 secretion from human epithelial cell lines. Suppression of IL-8 in human embryonic kidney (HEK293) cells that were transformed to express TLR5 was observed not only upon inflammatory stimulation by flagellin but also in response to tumor necrosis factor alpha (TNF-α) and phorbol myristate acetate (PMA), despite the fact that the TNF-α- and PMA-induced inflammatory pathways reportedly are not TLR5 mediated. SK1144 neither decreased IL-8 transcript accumulation nor increased intracellular retention of IL-8. No suppression was observed when the bacteria were cultured in Transwell cups above the epithelial cells; however, a nonadherent bacterial mutant (lacking the afimbrial adhesin gene) still inhibited IL-8 secretion. Direct contact between the bacteria and epithelial cells was necessary, but diffuse adhesion was dispensable for the inhibitory effects. Infection in the presence of chloramphenicol did not suppress cytokine release by the epithelial cells, suggesting that suppression depended on effectors synthesized de novo Inflammatory suppression was attenuated with infection by a bacterial mutant deleted for hcp (encoding a component of a type VI secretion system). In conclusion, DAEC strains from healthy carriers impede epithelial cell cytokine secretion, possibly by interfering with translation via the type VI secretion system.

Virulence gene profiles and molecular genetic characteristics of diarrheagenic Escherichia coli from a hospital in western China

BackgroundDiarrheagenic Escherichia coli (DEC) is one of the most important etiological agents of diarrheal diseases. In this study we investigated the prevalence, virulence gene profiles, antimicrobial resistance, and molecular genetic characteristics of DEC at a hospital in western China.MethodsA total of 110 Escherichia coli clinical isolates were collected from the First Affiliated Hospital of Chengdu Medical College from 2015 to 2016. Microbiological methods, PCR, antimicrobial susceptibility test, pulsed-field gel electrophoresis and multilocus sequence typing were used in this study.ResultsMolecular analysis of six DEC pathotype marker genes showed that 13 of the 110 E. coli isolates (11.82%) were DEC including nine (8.18%) diffusely adherent Escherichia coli (DAEC) and four (3.64%) enteroaggregative Escherichia coli (EAEC). The adherence genes fimC and fimH were present in all DAEC and EAEC isolates. All nine DAEC isolates harbored the virulence genes fyuA and irp2 and four (44.44%) also carried the hlyA and sat genes. The virulence genes fyuA, irp2, cnf1, hlyA, and sat were found in 100%, 100%, 75%, 50%, and 50% of EAEC isolates, respectively. In addition, all DEC isolates were multidrug resistant and had high frequencies of antimicrobial resistance. Molecular genetic characterization showed that the 13 DEC isolates were divided into 11 pulsed-field gel electrophoresis patterns and 10 sequence types.ConclusionsTo the best of our knowledge, this study provides the first report of DEC, including DAEC and EAEC, in western China. Our analyses identified the virulence genes present in E. coli from a hospital indicating their role in the isolated DEC strains’ pathogenesis. At the same time, the analyses revealed, the antimicrobial resistance pattern of the DEC isolates. Thus, DAEC and EAEC among the DEC strains should be considered a significant risk to humans in western China due to their evolved pathogenicity and antimicrobial resistance pattern.

Enteroaggregative Escherichia coli is the predominant diarrheagenic E. coli pathotype among irrigation water and food sources in South Africa

Diarrheagenic E. coli (DEC) has been implicated in foodborne outbreaks worldwide and have been associated with childhood stunting in the absence of diarrhoea. Infection is extraordinarily common, but the routes of transmission have not been determined. Therefore, determining the most prevalent pathotypes in food and environmental sources may help provide better guidance to various stakeholders in ensuring food safety and public health and advancing understanding of the epidemiology of enteric disease. We characterized 205 E. coli strains previously isolated from producer distributor bulk milk (PDBM)(118), irrigation water (48), irrigated lettuce (29) and street vendor coleslaw (10) in South Africa. Enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroaggregative E. coli (EAEC) and diffusely adherent E. coli (DAEC) were sought. We used PCR and partial gene sequencing for all 205 strains while 46 out of 205 that showed poor resolution were subsequently characterized using cell adherence (HeLa cells).PCR and partial gene sequencing of aatA and/or aaiC genes confirmed EAEC (2%, 5 out of 205) as the only pathotype. Phylogenetic analysis of sequenced EAEC strains with E. coli strains in GenBank showing ≥80% nucleotide sequence similarity based on possession of aaiC and aatA generated distinct clusters of strains separated predominantly based on their source of isolation (food source or human stool) suggesting a potential role of virulence genes in source tracking. EAEC 24%, 11 out of 46 strains (PDBM = 15%, irrigation water = 7%, irrigated lettuce = 2%) was similarly the predominant pathotype followed by strains showing invasiveness to HeLa cells, 4%, 2 out of 46 (PDBM = 2%, irrigated lettuce = 2%), among stains characterized using cell adherence.Therefore, EAEC may be the leading cause of DEC associated food and water-borne enteric infection in South Africa. Additionally, solely using molecular based methods targeting virulence gene determinants may underestimate prevalence, especially among heterogeneous pathogens such as EAEC.

High prevalence of diarrheagenic Escherichia coli carrying toxin-encoding genes isolated from children and adults in southeastern Brazil

BackgroundDiarrheagenic Escherichia coli (DEC) are important bacterial causes of childhood diarrhea in Brazil, but its impact in adults is unknown. This study aimed at investigating DEC among children and adults living in endemic areas.MethodsA total of 327 stools specimens were collected from children (n = 141) and adults (n = 186) with diarrhea attending health centers. Diarrheagenic E. coli (DEC) were identified by their virulence genes (multiplex polymerase chain reaction) and HEp-2 cell adherence patterns.ResultsDEC were detected in 56 (40%) children and 74 (39%) adults; enteroaggregative E. coli (EAEC) (23%) was the most prevalent pathotype, followed by diffusely adherent E. coli (DAEC) (13%), and occurred at similar frequencies in both diarrheal groups. Atypical enteropathogenic E. coli (aEPEC) strains were recovered more frequently from children (6%) than from adults (1%). Twenty-six percent of the EAEC were classified as typical EAEC possessing aggR gene, and carried the aap gene. EAEC strains carrying aggR-aap-aatA genes were significantly more frequent among children than adults (p < 0.05). DAEC strains possessing Afa/Dr. genes were detected from children (10%) and adults (6%). EAEC and DAEC strains harboring genes for the EAST1 (astA), Pet, Pic, and Sat toxins were common in both diarrheal groups. The astA and the porcine AE/associated adhesin (paa) genes were found in most of aEPEC strains. High levels of resistance to antimicrobial drugs were found among DAEC and aEPEC isolates.ConclusionThe results show a high proportion of EAEC and DAEC carrying toxin-encoding genes among adults with diarrhea.

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

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

Bacterial pathogens of pigs with particular reference to Escherichia coli: A systematic review and meta-analysis

Pigs are ungulate animals of the genus Suis and family Suidae. They are globally spread but restricted in certain countries due to religious and cultural beliefs. Pork serves as an important source of protein (38% of meat consumed in the world). While pig production remains a profitable enterprise, commercial and particularly the small-scale farmers face huge constraint in this husbandry practice, one of the most important being bacterial infections and its associated with morbidity and mortality. In this work, we reviewed the prevalence of bacterial infections in pigs with particular reference to Escherichia coli, a bacterium that is regularly isolated and can lead to multiple infections in pigs. Literatures were searched on selected veterinary and biological data bases in 2016 with focus on natural infections and isolates from natural infections with epidemiological details. Pathotypes, serotypes and serogroups of E. coli, the country of origin, source, growth stage, age of pigs infected, disease outbreak, the number of samples and type of samples, numbers and percentage of positive samples and isolates were used as filters. Pathotypes reported include enterotoxigenic E. coli (ETEC) 66.7%, enterotoxigenic E. coli and shiga toxigenic E. coli (ETEC and STEC) 14.3%, STEC only (7.9%), enterotoxigenic E. coli/enteropathogenic E. coli/enteroaggregative E. coli (ETEC/EPEC/EAE) 31.7%. Others were enterohaemorrhagic E. coli (EHEC), diffusely adherent E. coli (DAEC) (ETEC, EPEC, STEC) and extra-intestinal pathogenic E. coli (ExPEC). Twenty-nine countries with documented records of cases of E. coli were included with the USA reporting, the highest number followed by China. About 74% of the samples were taken from farms and others were from samples submitted to research laboratories and veterinary faculties for necropsy. Serogroups O141, O149, O139, O138, O8 and O9 were most common. Piglets were most affected (52.3%) followed by weaners (39.6%) and porkers (7.9%) with age ranging from 1 to 392 days old. A total of 24,854 isolates were considered, 10477 (42.2%) were positives and the following genes were carried: STa, STb, LT, stx1, stx2, Stx-2e, F4, F5, F6, F18, F41, AIDA, EAST1, eae, paa and hlyA. The diseases produced by E. coli were neonatal diarrhoea, colibacillosis, post-weaning diarrhoea and edema disease. The associated risk factors were poor housing, management and feed changes, extensive use of antibiotics as prophylaxis, overcrowding, and high humidity and temperature changes. India, USA, Japan, Slovakia, Denmark Sweden and Poland were countries with significant reports and high detection of virulence factors (72 to 100%). Key words: Escherichia coli, diarrhea, serogroups, enterotoxigenic, colibacillosis.