Coli Colonization Research Articles

Fecal Carriage of Colibactin-Encoding Escherichia coli Associated With Colorectal Cancer Among a Student Populace.

Fecal carriage of the colibactin (clb) gene in Escherichia coli is described as a source that could promote carcinogenesis, progressing to colorectal cancer. The present study investigated the demographic, dietary, and antibiotic consumption variables as correlates for fecal carriage of clb+/E coli among the student populace. In a randomized cross-sectional survey, E coli (N = 136) from the fecal samples of eligible students were characterized and evaluated for antibiotic resistance, β-lactamase (blm), biofilm, virulence factor production, and strain tryptophan reverse mutagenic activity. The encoded clb+/E coli were analyzed for correlates with principal component analysis. Of all the E coli strains, a low rate of 2 clb+/E coli (1.5%) and higher rates of biofilm (13.2%) and blm producers (11.8%) were recorded among the mutant strains as compared with the nonmutant types. All the clb+/E coli showed complete resistance to amoxicillin, Augmentin (amoxicillin and clavulanate), gentamicin, and trimethoprim/sulfamethoxazole. The fecal clb-encoded E coli (1.5%) were not associated with demographic status, fiber-based food (odds ratio [OR], 1.03; 95% CI, 56.74-138.7; P = .213), alcohol (OR, 1.27; 95% CI, 61.74-147.1; P = .221), antibiotic consumptions (OR, 1.11; 95% CI, 61.29-145.3; P = .222), and handwashing (OR, 1.17; 95% CI, 60.19-145.5; P = .216). The hierarchical cluster of blm+/E coli revealed high-level resistance with a multiantibiotic resistance index ≥0.2 (P < .05). Only 12% of all strains were tryptophan mutant/blm+, and 1.5% of clb+/ECblm+ were observed in fecal samples with a 452-base pair size. Trimethoprim/sulfamethoxazole and biofilm production positively regressed with clb expression (P > .05). Principal component analysis score plot indicated an association of clb+/ECblm+ with dietary pattern, alcohol, blm, and hemolysin production. The combined activity of blm and biofilm production in the gut microbiota could promote clb+/E coli colonization, facilitating genotoxin production and possible colorectal cancer induction.

Metagenome of Gut Microbiota Provides a Novel Insight into the Pathogenicity of Balantioides coli in Weaned Piglets

Balantioides coli plays an important role in the diarrhea of weaned piglets, but its pathogenic potential and interaction with gut microbes remain unclear. To investigate the impact of B. coli colonization on the gut bacterial structure and function of weaned piglets, a metagenomic analysis based on shotgun sequencing was performed on fresh fecal samples collected from ten B. coli-colonized piglets and eight B. coli-free ones in this study. The results showed that decreasing diversity and shifted composition and function of the bacterial community were detected in the weaned piglets infected by B. coli. In contrast to the B. coli-negative group, the relative abundances of some members of the Firmicutes phylum including Clostridium, Ruminococcus species, and Intestinimonas butyriciproducens, which produce short-chain fatty acids, were significantly reduced in the B. coli-positive group. Notably, some species of the Prevotella genus (such as Prevotella sp. CAG:604 and Prevotella stercorea) were significantly increased in abundance in the B. coli-positive piglets. A functional analysis of the gut microbiota demonstrated that the differential gene sets for the metabolism of carbohydrates and amino acids were abundant in both groups, and the more enriched pathways in B. coli-infected piglets were associated with the sugar-specific phosphotransferase system (PTS) and the two-component regulatory system, as well as lipopolysaccharide (LPS) biosynthesis. Furthermore, several species of Prevotella were significantly positively correlated to the synthesis of lipid A, leading to the exporting of endotoxins and, thereby, inducing inflammation in the intestines of weaned piglets. Taken together, these findings revealed that colonization by B. coli was distinctly associated with the dysbiosis of gut bacterial structure and function in weaned piglets. Lower relative abundances of Clostridiaceae and Ruminococcaceae and higher abundances of Prevotella species were biomarkers of B. coli infection in weaned piglets.

The effect of Campylobacter jejuni and Campylobacter coli colonization on the gut morphology, functional integrity, and microbiota composition of female turkeys

BackgroundCampylobacter (C.) species are the most common bacterial cause of foodborne diarrhea in humans. Despite colonization, most animals do not show clinical signs, making recognition of affected flocks and disruption of the infection chain before slaughter challenging. Turkeys are often cocolonized with C. jejuni and C. coli. To understand the pathogen-host-interaction in the context of two different Campylobacter species, we compared the colonization patterns and quantities in mono- and co-colonized female commercial turkeys. In three repeated experiments we investigated the impact on gut morphology, functional integrity, and microbiota composition as parameters of gut health at seven, 14, and 28 days post-inoculation.ResultsDespite successful Campylobacter colonization, clinical signs or pathological lesions were not observed. C. coli persistently colonized the distal intestinal tract and at a higher load compared to C. jejuni. Both strains were isolated from livers and spleens, occurring more frequently in C. jejuni- and co-inoculated turkeys. Especially in C. jejuni-positive animals, translocation was accompanied by local heterophil infiltration, villus blunting, and shallower crypts. Increased permeability and lower electrogenic ion transport of the cecal mucosa were also observed. A lower relative abundance of Clostridia UCG-014, Lachnospiraceae, and Lactobacillaceae was noted in all inoculated groups compared to controls.ConclusionsIn sum, C. jejuni affects gut health and may interfere with productivity in turkeys. Despite a higher cecal load, the impact of C. coli on investigated parameters was less pronounced. Interestingly, gut morphology and functional integrity were also less affected in co-inoculated animals while the C. jejuni load decreased over time, suggesting C. coli may outcompete C. jejuni. Since a microbiota shift was observed in all inoculated groups, future Campylobacter intervention strategies may involve stabilization of the gut microbiota, making it more resilient to Campylobacter colonization in the first place.

70274 TL1 team approach to investigating the adhesin gene fimH in adherent invasive E. coli induced inflammation and colorectal cancer development

ABSTRACT IMPACT: We are developing the 3D perfusion system for use with patient-derived bacteria to further characterize the mechanism behind bacterial-induced inflammation and cancer. OBJECTIVES/GOALS: We previously reported the adherent invasive E. coli NC101 promote colorectal cancer (CRC) in mice. FimH, a mannose-specific adhesin on type 1 fimbriae, is involved in bacterial surface adhesion. Herein, we investigated the role of FimH in E. coli NC101-induced adherence and carcinogenesis in a novel 3D perfusion culture imaging plate. METHODS/STUDY POPULATION: E. coli NC101 gene fimH was deleted byï ŲRed Recombinase System. Biofilm formation was assessed by crystal violet and congo red staining. 5 dpf (wild-type strain) zebrafish embryos were infected in 6x107 cfu/ml wild type (WT) or fimH-deleted (ï ,,fimH) E. coli NC101 for 24hr and gut dissected for bacterial culture. A 2D/3D infection culture system for IEC-6 and HT-29 cells was infected for 4 hr and imaged and then DNA damage examined by comet assay, cell cycle andÎ3H2AX accumulation. Germ-free (GF) Il10-/- (colitis) mice were orally gavaged with 108 cfu WT orï ,,fimH E. coli NC101 for 16 weeks. E. coli colonization were quantified by plate culture and qPCR. Lipocalin2 was quantified by ELISA. PCNA and β-catenin were evaluated by immunohistochemistry (IHC). RESULTS/ANTICIPATED RESULTS: Biofilm formation was reduced by more than 40% (p&lt;0.05) in E. coli NC101ï ,,fimH compared to WT strain. Zebrafish larvae showed a 41% decrease in intestinal colonization ofï ,,fimH compared to WT (p&lt;0.05). E. coli NC101-induced DNA damage was reduced by 67% (p&lt;0.0001) in HT-29 cells infected withï ,,fimH compared to WT strain. Using the 3D infection system, a 46% decrease in yH2AX (p&lt;0.05) and 42% decrease in G2 cell cycle arrest (p&lt;0.05) was observed inï ,,fimH infected IEC-6 cells compared to WT strain. Furthermore, ï ,,fimH infected Il10-/- mice showed decreased colonization (p&lt;0.01), decreased intestinal inflammation (p&lt;0.05), decreased stool lipocalin2 level (p&lt;0.01), and reduction of PCNA positive cells in the intestine (p&lt;0.05) compared to mice infected with WT strain. DISCUSSION/SIGNIFICANCE OF FINDINGS: Adhesin protein FimH is required by E. coli NC101 to colonize and promote colitis and carcinogenesis both in a 3D perfusion culture and in mice and may serve as potential therapeutic target.

Investigation of Campylobacter colonization in three Australian commercial free-range broiler farms

Campylobacter spp. contaminated poultry products are strongly associated with foodborne illnesses worldwide. Development of effective management strategies to reduce contamination by Campylobacter spp. requires an improved understanding of the numerous factors that drive these contamination processes. Currently, chicken farms are using more free-range chicken meat production systems in response to consumer preferences. However, Campylobacter spp. colonization has rarely been investigated on free-range broiler farms. The present study investigated the temporal and environmental factors influencing Campylobacter spp. colonization of free-range broilers as well as potential sources and genetic diversity of Campylobacter jejuni (C. jejuni) and Campylobacter coli (C. coli) in commercial free-range broiler farms. Genetic linkages among the isolates were analyzed using flaA amplicon analysis. Campylobacter coli was first detected in fecal samples of a commercial free-range broiler flock on day 10 of rearing. Multiple genotypes of C. jejuni and C. coli were identified in this study. The farm environment was identified as a potential source of C. jejuni and C. coli colonization of free-range broilers. The dominant Campylobacter genotype varied between free-range broiler farms over time, with C. jejuni being the most frequently isolated species. These findings enhance the understanding of C. jejuni and C. coli colonization in free-range broiler farms and could inform the development of more effective intervention strategies to help control this important foodborne pathogen.

Impact of Bacteriophage-Supplemented Drinking Water on the E. coli Population in the Chicken Gut.

Among intestinal coliform microbes in the broiler gut, there are potentially pathogenic Escherichia (E.) coli that can cause avian colibacillosis. The treatment with antibiotics favors the selection of multidrug-resistant bacteria and an alternative to this treatment is urgently required. A chicken model of intestinal colonization with an apathogenic model strain of E. coli was used to test if oral phage application can prevent or reduce the gut colonization of extraintestinal pathogenic E. coli variants in two individual experiments. The E. coli strain E28 was used as a model strain, which could be differentiated from other E. coli strains colonizing the broiler gut, and was susceptible to all cocktail phages applied. In the first trial, a mixture of six phages was continuously applied via drinking water. No reduction of the model E. coli strain E28 occurred, but phage replication could be demonstrated. In the second trial, the applied mixture was limited to the four phages, which showed highest efficacy in vitro. E. coli colonization was reduced in this trial, but again, no reduction of the E. coli strain E28 was observed. The results of the trials presented here can improve the understanding of the effect of phages on single strains in the multi-strain microbiota of the chicken gut.

The response of nitrogen cycling and bacterial communities to E. coli invasion in aquatic environments with submerged vegetation

The effects of exogenous Escherichia coli on nitrogen cycling (N-cycling) in freshwater remains unclear. Thus, seven ecosystems, six with submerged plants-Potamogeton crispus (PC) and Myriophyllum aquaticum (MA)-and one with no plants were set up. Habitats were assessed before and after E. coli addition (107colony-forming units/mL). E. coli colonization of freshwater ecosystems had significant effects on bacterial community structure in plant surface biofilms and surface sediments (ANOVA, P<0.05). It reduced the relative abundance of nitrosification bacteria (-70.94±26.17%) and nitrifiers (-47.86±23.68%) in biofilms which lead to significant reduction of ammoxidation in water (P<0.05). The N-cycling intensity from PC systems was affected more strongly by E. coli than were MA systems. Furthermore, the coupling coefficient of exogenous E. coli to indigenous N-cycling bacteria in sediments (6.061, average connectivity degree) was significantly weaker than that in biofilms (9.852). Additionally, at the genus level, E. coli were most-closely associated with N-cycling bacteria such as Prosthecobacter, Hydrogenophaga, and Bacillus in sediments and biofilms according to co-occurrence bacterial network (Spearman). E. coli directly changed their abundance, so that the variability of species composition of N-cycling bacterial taxa was triggered, as well. Overall, exogenous E. coli repressed ammoxidation, but promoted ammonification and denitrification. Our results provided new insights into how pathogens influence the nitrogen cycle in freshwater ecosystems.

Enterotoxigenic Escherichia Coli Toxins and Colonization Factors Among Zambian Children Presenting with Moderate to Severe Diarrhea to Selected Health Facilities

Enterotoxigenic Escherichia Coli (ETEC) is an important cause for diarrheal disease in children and travelers globally. Epidemiological data on the distribution of strains of ETEC and associated colonization factors (CFs) is important for evaluation of candidate vaccines. We used conventional PCR and quantitative PCR to screen for toxins and CFs using DNA extracted from stool samples which tested positive for ETEC using the Luminex GPP panel collected from children presenting with moderate to severe diarrhea at selected health facilities in Lusaka. 49/106 (46.2%) were positive for at least one toxin (i.e. LT/STh/STp), ST was 18 (17%) [STh 16(15%) and STp 2 (~2%)], and LT 16(15%). The most frequent CF detected was CS6 with 6/49 (12.2%), followed by CS2, CS3 and CS7 with 2/49 (4.1%) each. CS6 was common across all toxin combinations (LT only, STh only and a combination of LT/STh) while CS2, CS3, CS7 were identified in both LT and LT/STh strains respectively. The mean age of children with detected toxin or CFs was 15.4 months (95% CI: 12.2, 18.7). Our results offer an insight into relevant CFs in ETEC diarrhea in Zambia and that Luminex™ platform is not as specific as ordinary and quantitative PCR for ETEC detection.

Incidence and Risk Factors for colonization of Multidrug-Resistance Organisms (MDROs) among Patients Undergoing Elective Orthopedic Surgery at Thammasat University Hospital

BackgroundThere is limited data on incidence of MDROs colonization and outcome in patients undergoing elective orthopedic surgery.MethodsWe performed MDROs surveillance screening (swabs from nose, throat, groins, and rectum) in patients undergoing elective orthopedic surgery at the Thammasat University Hospital between March and August 2016. MDROs were defined as Gram-negative bacteria (GNB) possessing extended-spectrum β-lactamases (ESBLs), Carbapenem-resistant Enterobacteriaceae (CRE), and non-lactose fermenting GNB resistant to at least 3 antibiotic classes, methicillin-resistant Staphylococcus aureus (MRSA), and vancomycin resistant enterococci (VRE). MDROs were identified by the Vitek®2 automated system. Antimicrobial susceptibility testing (disk diffusion test) was performed using the CLSI Interpretive Guidelines. Incidence of MDROs colonization upon admission was determined. Patient’s clinical characteristics, risk factors for MDROs infection, procedure types and antibiotic prophylaxis were prospectively collected. Surgical sites infections (SSIs) and complications up to 6 months after surgery among the patients with and without MDROs colonization were compared.ResultsOf 384 swabs tested from 96 patients (median age, 58 years), ESBL-producing Escherichia coli (ESBL-E. coli) was identified in 31 rectal swabs (32.3%) and 7 groin swabs (7.3%). Seven patients (7.3%) were diagnosed with SSIs. A higher rate of SSIs was found among patients with ESBL-E. coli colonization (6/31, 19.4%) compared with patient without ESBL-E. coli colonization (1/65, 1.5%; P = 0.004, OR 15.4, 95% CI 1.7–356.3). From the multivariate logistic regression analysis, SSIs were significantly associated with ESBL-E.coli colonization (P = 0.009, adjusted OR 18.3, 95% CI 2.1–162.9). In addition, from the multivariate logistic regression analysis, ESBL-E.coli was a significantly risk factor associated with SSIs (6/7, 85.7%, P = 0.014, adjusted OR 16.5, 95% CI 1.8–153.4).ConclusionWe found a high incidence of ESBL-E. coli colonization and rate of SSIs in patients who had elective orthopedic surgery. These patients did not have any other risk factors for MDROs infection. Active screening for colonization of ESBL-producing pathogens may be necessary to guide appropriate antibiotic prophylaxis to reduce rates of SSIs.Disclosures All authors: No reported disclosures.

Functional Characterization of EscK (Orf4), a Sorting Platform Component of the Enteropathogenic Escherichia coli Injectisome.

The type III secretion system (T3SS) is an essential virulence determinant for enteropathogenic Escherichia coli (EPEC) colonization of intestinal epithelial cells. Multiple EPEC effector proteins are injected via the T3SS into enterocyte cells, leading to diarrheal disease. The T3SS is encoded within a genomic pathogenicity island termed the locus of enterocyte effacement (LEE). Here we unravel the function of EscK, a previously uncharacterized LEE-encoded protein. We show that EscK is central for T3SS biogenesis and function. EscK forms a protein complex with EscQ, the main component of the cytoplasmic sorting platform, serving as a docking site for T3S substrates. Our results provide a comprehensive functional analysis of an understudied component of T3SSs.

Effects of dietary essential oil and enzyme supplementation on growth performance and gut health of broilers challenged by Clostridium perfringens

Abstract An experiment was carried out to evaluate the effects of dietary supplemental essential oil comprising thymol and carvacrol (EO), and enzyme complex containing xylanase, glucanase and mannanase as major components (CSM) on Clostridium perfringens challenged model in broilers. A total of 648 1-d-old female broiler chicks were assigned to eight groups. A 2 × 2 × 2 factorial arrangement of treatments was used in a randomized complete block design to study the effects of essential oil supplementation (with or without EO 60 mg/kg diet), enzymes addition (with or without CSM 500 mg/kg diet), pathogen challenge (with or without C. perfringens challenge from d 14 to 20), and the interactive effects. A pelleted high NSP wheat–soybean meal basal diet was formulated to meet nutrient requirements recommended by NRC (1994) . Body weight, feed intake, and feed to gain ratio were determined on d 14, 21 and 35; ileal digesta viscosity, intestinal lesion score, ileal histomorphologic indices, bacterial population and plasma endotoxin were measured on d 21; while ileal permeability was determined from d 24 to 26. Results indicated that enzyme CSM supplementation increased body weight on d 21 and 35, improved feed to gain ratio from d 0 to 14 and 0 to 21, increased P-glycoprotein dependent efflux permeability ( P app (S–M)), decreased digesta viscosity, and lowered ileal C. perfringens and Lactobacillus colonization. CSM supplementation could also improve feed conversion ratio from d 0 to 35 and VH/CD on d 21 of C. perfringens challenged chicks, while EO supplementation increased villus height and VH/CD. C. perfringens challenge increased intestinal lesion score, C. perfringens and Lactobacillus colonization, but decreased Bifidobacterium colonization and P app (S–M). C. perfringens challenge also resulted in increased plasma endotoxin, passive transcellular permeability ( P app (M–S)), sIgA, and E. coli colonization when chicks were fed with basal diet, while EO or CSM supplementation could alleviate the alteration caused by C. perfringens infection. In conclusion, dietary enzyme CSM supplementation could benefit for growth performance and gut health, and EO could improve the gut health of C. perfringens challenged chicks.

Prevalence of fecal carriage of extended-spectrum- and metallo-β-lactamase-producing gram-negative bacteria among neonates born in a hospital setting in central Saudi Arabia.

BACKGROUND AND OBJECTIVECommensal neonatal fecal flora constitute a reservoir of antibiotic resistance. The aim of this study was to characterize the prevalence of fecal carriage of extended spectrum beta-lactamases (ESBLs) and carbapenemase producing gram-negative bacteria among 150 neonates who were born in two hospitals in central Saudi Arabia.PATIENTS AND METHODSFrom June 2012 to Janauary 2013, 150 healthy neonates ≤7 days of age were screened for intestinal enteric bacteria colonization and the influence of age, mode of delivery, body weight and breast feeding of neonates on colonzation rates. The isolates were identified manually and by VITEK 2 and antibiotic susceptibility was tested. Phenotypic tests were performed to seek ESBL and metallo-β-lactamase (MBL) producers in recovered isolates.RESULTSAmong 150 fecal specimens, 188 gram-negative rods were recovered. E coli (n=130; 69.1%) was the commonest organism followed by Klebsiella pneumoniae (n=23; 12.2%), Pseudomonas aeuroginosa (n=14; 7.5%), Acinetobacter baumannii (n=9; 4.8%), Enterobacter spp. (n=9; 4.8%) and Morganella morganii (n=3; 1.6%). E coli colonization during the first day of life was 95.5% (n=43/45), which was relatively higher than in subsequent days. The colonization rate of E coli among low weight babies (≤3 kg) was significantly higher than fully grown neonates (≥4 kg)(P=.03). The prevalence of ESBL and MBL was 15.0% (n=28/188) and 1.6% (n=3/188). The prevalence rates of ESBL in E coli, K pneumoniae, E cloacae, M morganii and E aerogenes were 16.9% (n=22/130), 4.3 % (n=1/23), 60% (n=3/5), 33.3% (n=1/3), and 25% (n=1/4), respectively. All E cloacae with ESBLs and MBLs were resistant to all tested antibiotics including carbapenems.CONCLUSIONWithin the first week of life, the most important determinants of gut enteric bacteria composition in neonates were age in days, body weight, mode of delivery, and type of neonatal feeding. The findings of ESBLs- and MBLs-producing enteric bacteria as neonate-gut colonizers may have wider implication or linkage to prevention of nosocomial and community-acquired infections by these strains.

Enteropathogenic Escherichia coli (EPEC): Does it have a role in colorectal tumourigenesis? A Prospective Cohort Study

BackgroundDespite the characterization of many aetiologic genetic changes. The specific causative factors in the development of sporadic colorectal cancer remain unclear. This study was performed to detect the possible role of Enteropathogenic Escherichia coli (EPEC) in developing colorectal carcinoma. Patients and methodFresh biopsy specimens have been obtained from the colonic mucosa overlying the colorectal cancer as well as from the colon of the healthy controls. Culture, genotyping and virulence of EPEC were done using (nutrient broth culture, and PCR). Strains biochemically identified as Escherichia coli were selected from the surface of a MacConkey's plate and were serogrouped by slide agglutination tests. ResultsFrom January 2011 to June 2014, 213 colorectal cancer patients (Group 1) and 248 healthy controls (Group 2) were prospectively enrolled in this study. EPEC was positive in 108 (50.7%) in group 1 and 51 (20.6%) in group 2 (P = 0.0001). A significant difference between both groups was observed regarding serotyping, genotyping (eae gene) and virulence category (P = 0.0001). A significant difference between the 2 subgroups of colorectal cancer cases was observed regarding genotyping (eae, bfb genes) and virulence category. ConclusionThe incidence EPEC was higher significantly in patients with colorectal cancer. E. coli in patients with colorectal cancer significantly differed serotypically and genotypically from the E. coli in normal population. E. coli colonization of the colonic mucosa may be a cause colorectal cancer.

Epidermal growth factor improved alcohol-induced inflammation in rats

The purpose of this study was to investigate the effects of an epidermal growth factor (EGF) intervention on improving the inflammatory response of rats fed an ethanol-containing diet. Eight-week-old male Wistar rats were divided into ethanol (E) and control (C) groups. Rats in the E group were fed an ethanol liquid diet, while rats in the C group were pair-fed an isoenergetic diet without ethanol. After a 4-week ethanol-induction period, both the C and E group were respectively subdivided into 2 groups: a normal liquid diet without (C group, n = 8) or with EGF supplementation (C + EGF, n = 8), and the ethanol-containing diet without (E group, n = 8) or with EGF supplementation (E + EGF group, n = 8). The EGF (30 μg/kg body weight/day) intervention period was carried out for the following 8 weeks. At the end of the experiment, activity of aspartate transaminase (AST) and alanine transaminase (ALT) and hepatic levels of tumor necrosis factor (TNF)-α, interleukin (IL)-1β, IL-6, and IL-10 in group E were significantly higher than those in group C. In addition, alterations in the gut microbiota profile were found in group E. In contrast, activity of AST and ALT and levels of TNF-α, IL-1β, and IL-6 in group E + EGF were significantly lower than those in group E. Significantly lower intestinal permeability and lower numbers of Escherichia coli in the fecal microbial culture were also found in group E + EGF. These results suggest that EGF improved the intestinal integrity by decreasing E. coli colonization and lowering intestinal permeability, which then ameliorated the inflammatory response under chronic ethanol exposure.

Differences in the gut microbiota of healthy children and those with type 1 diabetes

Intestinal barriers, intestinal flora, and mucosal immunity are the main factors responsible for the development of various allergic and autoimmune diseases. The aim of this study was to investigate the relationship between the intestinal flora of children and the presence of type 1 diabetes, and to determine if gut microbiota could partly explain the etiology of the disease. Fecal flora analysis was done using quantitative cultures on selective and non-selective media with different thermal and atmospheric conditions for bacterial and fungal growth. The study group consisted of 35 patients (16 female, 19 male; mean age, 10.73 ± 4.16 years), who had been followed by the University of Istanbul, Cerrahpasa Medical Faculty, Department of Pediatrics, and were newly diagnosed with type 1 diabetes. The control group consisted of 35 healthy subjects (15 female, 20 male; mean age, 9.96 ± 4.09 years), who were randomly selected and had similar demographics. Bifidobacterium colonization was lower in patients with type 1 diabetes compared to the control group, whereas Candida albicans and Enterobacteriaceae other than Echerichia coli colonization was increased. A decrease in beneficial anaerobic bacteria levels and a concomitant increase in Enterobacteriaceae other than E. coli and C. albicans colonization may lead to a disturbance in the ecological balance of intestinal flora, which could be a triggering factor in type 1 diabetes etiology.

Risk Factors Associated with the Community-Acquired Colonization of Extended-Spectrum Beta-Lactamase (ESBL) Positive Escherichia Coli. An Exploratory Case-Control Study

BackgroundThe number of extended-spectrum beta-lactamase (ESBL) positive (+) Escherichia coli is increasing worldwide. In contrast with many other multidrug-resistant bacteria, it is suspected that they predominantly spread within the community. The objective of this study was to assess factors associated with community-acquired colonization of ESBL (+) E. coli.MethodsWe performed a matched case-control study at the Charité University Hospital Berlin between May 2011 and January 2012. Cases were defined as patients colonized with community-acquired ESBL (+) E. coli identified <72 h after hospital admission. Controls were patients that carried no ESBL-positive bacteria but an ESBL-negative E.coli identified <72 h after hospital admission. Two controls per case were chosen from potential controls according to admission date. Case and control patients completed a questionnaire assessing nutritional habits, travel habits, household situation and language most commonly spoken at home (mother tongue). An additional rectal swab was obtained together with the questionnaire to verify colonization status. Genotypes of ESBL (+) E. coli strains were determined by PCR and sequencing. Risk factors associated with ESBL (+) E. coli colonization were analyzed by a multivariable conditional logistic regression analysis.ResultsWe analyzed 85 cases and 170 controls, respectively. In the multivariable analysis, speaking an Asian language most commonly at home (OR = 13.4, CI 95% 3.3–53.8; p<0.001) and frequently eating pork (≥3 meals per week) showed to be independently associated with ESBL colonization (OR = 3.5, CI 95% 1.8–6.6; p<0.001). The most common ESBL genotypes were CTX-M-1 with 44% (n = 37), CTX-M-15 with 28% (n = 24) and CTX-M-14 with 13% (n = 11).ConclusionAn Asian mother tongue and frequently consuming certain types of meat like pork can be independently associated with the colonization of ESBL-positive bacteria. We found neither frequent consumption of poultry nor previous use of antibiotics to be associated with ESBL colonization.

Cefpodoxime vs Ciprofloxacin for Short-Course Treatment of Acute Uncomplicated Cystitis

Although fluoroquinolones remain the most reliable urinary antimicrobial, resistance rates have increased and effective fluoroquinolone-sparing antimicrobials are needed. To determine whether cefpodoxime is noninferior to ciprofloxacin for treatment of acute cystitis. Randomized, double-blind trial of 300 women aged 18 to 55 years with acute uncomplicated cystitis comparing ciprofloxacin (n = 150) with cefpodoxime (n = 150); patients were from a student health center in Seattle, Washington, and a referral center in Miami, Florida. The study was conducted from 2005 to 2009 and outcomes were assessed at 5 to 9 days and 28 to 30 days after completion of therapy. Intent-to-treat and per-protocol analyses were performed; 15 women in the ciprofloxacin group and 17 women in the cefpodoxime group were lost to follow-up. Patients were given 250 mg of ciprofloxacin orally twice daily for 3 days or 100 mg of cefpodoxime proxetil orally twice daily for 3 days. Overall clinical cure (defined as not requiring antimicrobial treatment during follow-up) at the 30-day follow-up visit. Secondary outcomes were clinical and microbiological cure at the first follow-up visit and vaginal Escherichia coli colonization at each follow-up visit. The hypothesis that cefpodoxime would be noninferior to ciprofloxacin by a 10% margin (ie, for the difference in the primary outcome for ciprofloxacin minus cefpodoxime, the upper limit of the confidence interval would be <10%) was formulated prior to data collection. The overall clinical cure rate at the 30-day visit with the intent-to-treat approach in which patients lost to follow-up were considered as having clinical cure was 93% (139/150) for ciprofloxacin compared with 82% (123/150) for cefpodoxime (difference of 11%; 95% CI, 3%-18%); and for the intent-to-treat approach in which patients lost to follow-up were considered as having not responded to treatment, the clinical cure rate was 83% (124/150) for ciprofloxacin compared with 71% (106/150) for cefpodoxime (difference of 12%; 95% CI, 3%-21%). The microbiological cure rate was 96% (123/128) for ciprofloxacin compared with 81% (104/129) for cefpodoxime (difference of 15%; 95% CI, 8%-23%). At first follow-up, 16% of women in the ciprofloxacin group compared with 40% of women in the cefpodoxime group had vaginal E coli colonization. Among women with uncomplicated cystitis, a 3-day regimen of cefpodoxime compared with ciprofloxacin did not meet criteria for noninferiority for achieving clinical cure. These findings, along with concerns about possible adverse ecological effects associated with other broad-spectrum β-lactams, do not support the use of cefpodoxime as a first-line fluoroquinolone-sparing antimicrobial for acute uncomplicated cystitis. clinicaltrials.gov Identifier: NCT00194532.

Micropatterned Surfaces for Reducing the Risk of Catheter-Associated Urinary Tract Infection: AnIn VitroStudy on the Effect of Sharklet Micropatterned Surfaces to Inhibit Bacterial Colonization and Migration of UropathogenicEscherichia coli

Catheter-associated urinary tract infection (CAUTI) is the most common device-associated infection and can result in serious medical consequences. We studied the efficacy of a novel microscopic physical surface modification (Sharklet) for preventing bacterial colonization and migration of uropathogenic Escherichia coli on silicone elastomer. In vitro growth assays evaluated E coli colonization using three variations of micropatterned silicone surfaces vs a smooth silicone control. Enumeration techniques included quantification of colonies on surfaces and analysis of bacterial area coverage and colony size. In vitro migration assays involved placement of micropatterned and smooth silicone rod segments between two agar islands to measure incidence of migration. All three variations of the Sharklet micropattern outperformed the control surfaces in inhibiting E coli colonization. On average, 47% reduction in colony-forming units (CFUs) and bacterial area coverage plus 77% reduction in colony size were achieved with the Sharklet surfaces in tryptic soy broth and artificial urine compared with the control nonpatterned surfaces. The incidence of E coli migration over the rod segments was reduced by more than 80% for the Sharklet transverse patterned rods compared with the unpatterned control rods. The Sharklet micropattern is effective at inhibiting colonization and migration of a common uropathogen. This performance is achieved through a physical surface modification without the use of any antimicrobial agents. Because deterrence of bacterial colonization and migration is a critical step to prevent CAUTI, the Sharklet micropattern offers a novel concept in addressing this important problem.

Risk factors for colonization of E. coli in Atlantic Bottlenose Dolphins (Tursiops truncatus) in the Indian River Lagoon, Florida.

Opportunistic pathogens related to degradation in water quality are of concern to both wildlife and public health. The objective of this study was to identify spatial, temporal, and environmental risk factors for E. coli colonization among Atlantic bottlenose dolphins (Tursiops truncatus) inhabiting the Indian River Lagoon (IRL), FL between 2003 and 2007. Age, gender, capture location, coastal human population density, proximity of sewage treatment plants, number of septic tanks, cumulative precipitation 48 hrs and 30 days prior to capture, salinity, and water temperature were analyzed as potential risk factors. Highest E. coli colonization rates occurred in the northern segments of the IRL. The risk of E. coli colonization was the highest among the youngest individuals, in counties with the highest cumulative rainfall 48 hrs and in counties with the highest number of septic systems during the year of capture. The prevalence of colonization was the highest during 2004, a year during which multiple hurricanes hit the coast of Florida. Septic tanks, in combination with weather-related events suggest a possible pathway for introduction of fecal coliforms into estuarine ecosystems. The ability of E. coli and related bacteria to act as primary pathogens or cause opportunistic infections adds importance of these findings.

Inhibition of bacterial adherence on the surface of biliary stent materials modified with chitosan

Bacterial infection plays an important role in the initiation of biliary sludge formation. Bacterial adherence and biofilm formation on the surface of a material have been considered as one of the main factors of stent re-occlusion in clinic. This work reported preventing bacterial adherence and bacterial biofilm formation on the surface of biliary stent material using chitosan film. The chitosan film was deposited on 316 L stainless steel (SS) plate by electrophoresis method and was characterized by X-ray diffraction (XRD), Fourier Transform infrared spectroscopy (FTIR) and atomic force microscopy (AFM). The ability of inhibiting bacterial adherence was investigated by incubating in human fresh bile adding E. coli and Enterobacter at 37±1 °C. Scanning electron microscopy (SEM) and fluorescence staining were used for observing bacterial colonization and biofilm formation. The results show that chitosan film was uniformly deposited on material surface, and the composition of the film did not change through cross-linking, but the crystallinity of chitosan film become well. Comparing to un-modified sample, the E. coli and Enterococcus adhesion amount and colonization on the surface of modified sample were significantly decreased by fluorescence staining and SEM. It is suggested that chitosan could be applied to biliary stent in clinical because of its antimicrobial activities.