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1

Uropathogenic Escherichia coli Induces Chronic Pelvic Pain ?  

PubMed Central

Chronic prostatitis/chronic pelvic pain syndrome (CP/CPPS) is a debilitating syndrome of unknown etiology often postulated, but not proven, to be associated with microbial infection of the prostate gland. We hypothesized that infection of the prostate by clinically relevant uropathogenic Escherichia coli (UPEC) can initiate and establish chronic pain. We utilized an E. coli strain newly isolated from a patient with CP/CPPS (strain CP1) and examined its molecular pathogenesis in cell culture and in a murine model of bacterial prostatitis. We found that CP1 is an atypical isolate distinct from most UPEC in its phylotype and virulence factor profile. CP1 adhered to, invaded, and proliferated within prostate epithelia and colonized the prostate and bladder of NOD and C57BL/6J mice. Using behavioral measures of pelvic pain, we showed that CP1 induced and sustained chronic pelvic pain in NOD mice, an attribute not exhibited by a clinical cystitis strain. Furthermore, pain was observed to persist even after bacterial clearance from genitourinary tissues. CP1 induced pelvic pain behavior exclusively in NOD mice and not in C57BL/6J mice, despite comparable levels of colonization and inflammation. Microbial infections can thus serve as initiating agents for chronic pelvic pain through mechanisms that are dependent on both the virulence of the bacterial strain and the genetic background of the host. PMID:21078846

Rudick, Charles N.; Berry, Ruth E.; Johnson, James R.; Johnston, Brian; Klumpp, David J.; Schaeffer, Anthony J.; Thumbikat, Praveen

2011-01-01

2

Virulence characteristics and antimicrobial susceptibility of uropathogenic Escherichia coli strains.  

PubMed

Eight virulence factors associated with uropathogenic Escherichia coli (UPEC) were investigated in 204 clinical isolates of E. coli recovered from urine cultures at counts ?10(5). The bacteria were classified into two groups according to the number of leukocytes in urine samples from which they were isolated: group I ?8 leukocytes/hpf, 104 strains; group II >8 leukocytes/hpf, 100 strains. Two multiplex PCR systems were used to detect genes encoding adhesin P (pap), adhesin S (sfa), afimbrial adhesin I (afa), siderophore aerobactin (aer), alpha-hemolysin (hly), cytotoxic necrotizing factor type 1 (cnf1), and traT associated with serum resistance. The PAI marker for the virulence island identified in strains CFT072 and CVD432, a marker of enteroaggregative E. coli, was also investigated using PCR. The susceptibility profile of E. coli strains was determined by disk diffusion method. Ninety percent UPEC showed at least one of the virulence genes, the prevalence being traT (76%), aer (41%), PAI (32%), sfa (26%), pap (25%), cnf1 (18%), afa (6%), and hly (5%). There was no significant difference in the distribution of virulence genes between groups I and II. A significantly higher degree of virulence was detected in UPEC group II. The CVD432 gene was not detected in any of the UPECs. Fifty-nine percent of the strains were resistant to at least one of the antimicrobials that we tested; the most common being resistance to ampicillin (51%) and trimethoprim-sulfamethoxazole (44%). PMID:22057993

Oliveira, F A; Paludo, K S; Arend, L N V S; Farah, S M S S; Pedrosa, F O; Souza, E M; Surek, M; Picheth, G; Fadel-Picheth, C M T

2011-01-01

3

Biofilm and fluoroquinolone resistance of canine Escherichia coli uropathogenic isolates  

PubMed Central

Background Escherichia coli is the most common uropathogen involved in urinary tract infection (UTI). Virulence of strains may differ, and may be enhanced by antimicrobial resistance and biofilm formation, resulting in increased morbidity and recurrent infections. The aim of this study was to evaluate the in vitro biofilm forming capacity of E. coli isolates from dogs with UTI, by using fluorescent in situ hybridization, and its association with virulence genes and antimicrobial resistance. Findings The proportion of biofilm-producing isolates significantly increased with the length of incubation time (P??0.05), but was significantly associated with afa, aer and the ?-lactamase genes (P?coli isolates from dogs with UTI. Biofilm formation may contribute to UTI treatment failure in dogs, through the development of bacterial reservoirs inside bladder cells, allowing them to overcome host immune defenses and to establish recurrent infections. PMID:25099929

2014-01-01

4

Functional analysis of the uropathogenic Escherichia coli R049 gene.  

PubMed

The objective of this study was to determine the function of the novel uropathogenic Escherichia coli (UPEC) gene R049 during host infection. We infected the urinary tracts of mice with E. coli UPEC132 or the R049 deletion mutant UPEC132?R049.The mouse kidneys were harvested at 4 and 8h post-infection and screened for differentially expressed genes by microarray analysis. We identified 379 and 515 differentially expressed genes at 4 and 8h post-infection, respectively. Thirty-four of these genes were associated with inflammatory and immune signaling pathways, including those related to mitogen-activated protein kinase signaling, leukocyte transendothelial migration, cytokine-cytokine receptor interaction, Toll-like receptor signaling, and apoptosis. Protein binding (GO 0005515) was the most prevalent molecular function in the Gene Ontology terms related to differentially expressed genes. In conclusion, R049 expression in UPEC132 is related to the early innate immune and inflammatory responses in UPEC-infected hosts. This work lays the foundation for further research on anti-infective immunity against UPEC. PMID:25644951

Yang, Dongjing; Dong, Jie; Su, Xu; Zhang, Wei; Zhang, Li; Li, Li; Lv, Likun; Guo, Liru

2015-02-01

5

Genomic islands of uropathogenic Escherichia coli contribute to virulence.  

PubMed

Uropathogenic Escherichia coli (UPEC) strain CFT073 contains 13 large genomic islands ranging in size from 32 kb to 123 kb. Eleven of these genomic islands were individually deleted from the genome, and nine isogenic mutants were tested for their ability to colonize the CBA/J mouse model of ascending urinary tract infection. Three genomic island mutants (Delta PAI-aspV, Delta PAI-metV, and Delta PAI-asnT) were significantly outcompeted by wild-type CFT073 in the bladders and/or kidneys following transurethral cochallenge (P coli strains but absent from E. coli K-12. We have shown that, in addition to encoding virulence genes, genomic islands contribute to the overall fitness of UPEC strain CFT073 in vivo. PMID:19329634

Lloyd, Amanda L; Henderson, Tiffany A; Vigil, Patrick D; Mobley, Harry L T

2009-06-01

6

Transposon Mutagenesis Identifies Uropathogenic Escherichia coli Biofilm Factors  

PubMed Central

Uropathogenic Escherichia coli (UPEC), which accounts for 85% of urinary tract infections (UTI), assembles biofilms in diverse environments, including the host. Besides forming biofilms on biotic surfaces and catheters, UPEC has evolved an intracellular pathogenic cascade that culminates in the formation of biofilm-like intracellular bacterial communities (IBCs) within bladder epithelial cells. Rapid bacterial replication during IBC formation augments a build-up in bacterial numbers and persistence within the host. Relatively little is known about factors mediating UPEC biofilm formation and how these overlap with IBC formation. To address this gap, we screened a UPEC transposon mutant library in three in vitro biofilm conditions: Luria broth (LB)-polyvinyl chloride (PVC), YESCA (yeast extract-Casamino Acids)-PVC, and YESCA-pellicle that are dependent on type 1 pili (LB) and curli (YESCA), respectively. Flagella are important in all three conditions. Mutants were identified that had biofilm defects in all three conditions but had no significant effects on the expression of type 1 pili, curli, or flagella. Thus, this approach uncovered a comprehensive inventory of novel effectors and regulators that are involved in UPEC biofilm formation under multiple conditions. A subset of these mutants was found to be dramatically attenuated and unable to form IBCs in a murine model of UTI. Collectively, this study expands our insights into UPEC multicellular behavior that may provide insights into IBC formation and virulence. PMID:22984258

Hadjifrangiskou, Maria; Gu, Alice P.; Pinkner, Jerome S.; Kostakioti, Maria; Zhang, Ellisa W.; Greene, Sarah E.

2012-01-01

7

Biofilm exclusion of uropathogenic bacteria by selected asymptomatic bacteriuria Escherichia coli strains  

Microsoft Academic Search

Many bacterial infections are associated with biofilm formation. In the urinary tract bacterial biofilms develop on both living surfaces and artificial implants, producing chronic and often intractable infections. Escherichia coli is the most common organism associated with urinary tract infections. In contrast to uropathogenic E. coli (UPEC), which cause symptomatic urinary tract infection, asymptomatic bacteriuria (ABU) strains are associated with

Lionel Ferrieres; Viktoria Hancock; Per Klemm

2007-01-01

8

Distribution of uropathogenic virulence factors among Escherichia coli strains isolated from dogs and cats.  

PubMed

A variety of virulence factors (VFs) such as type 1 fimbriae, pilus associated with pyelonephritis, S fimbriae, afimbrial adhesin, alpha-hemolysin, aerobactin and cytotoxic necrotizing factor 1 are associated with uropathogenic Escherichia coli. In this study, 80 uropathogenic E. coli strains in 50 dogs and 30 cats suffering from UTI. In addition, 60 E. coli strains were isolated from fecal samples from 30 each of healthy dogs and cats. The distribution of VFs of uropathogenic E. coli strains isolated from dogs and cats suffering from urinary tract infections (UTI) were examined by the colony hybridization test with seven DNA probes specific for VFs, and the results were compared with those obtained in the studies on strains from humans with UTI. In uropathogenic E. coli strains isolated from dogs and cats suffering from UTI, VFs were detected as frequently as in the strains isolated from humans with UTI. Although less frequently, genes encoding these VFs especially pap, sfa, hly, and cnf 1 genes were also associated with E. coli strains isolated from feces of healthy cats, in contrast to the distribution pattern of uropathogenic E. coli observed in humans. Furthermore, all VFs except pil were significantly more frequently detected in strains isolated from urine of animals with cystitis than in those isolated from feces of healthy humans. These results indicate that VFs of E. coli contribute to the pathogenesis of UTI in dogs and cats. PMID:9560773

Yuri, K; Nakata, K; Katae, H; Yamamoto, S; Hasegawa, A

1998-03-01

9

Multiple-Antibiotic Resistance Mediated by Plasmids and Integrons in Uropathogenic Escherichia coli and Klebsiella pneumoniae  

Microsoft Academic Search

Antibiotic resistance in urinary tract infection (UTI) is a growing public health problem in the world. In this study, a total of 182 uropathogens were isolated from patients with symptoms of urinary tract infection (UTI). Escherichia coli (88%) was the most prevalent isolate, while Klebsiella pneumoniae was recovered from 12% cases. The male\\/female ratio was 1:3. About 56% female and

Taslima Taher Lina; Sabita Rezwana Rahman; Donald James Gomes

2007-01-01

10

Integrin-Mediated Host Cell Invasion by Type 1–Piliated Uropathogenic Escherichia coli  

Microsoft Academic Search

Uropathogenic Escherichia coli (UPEC), the primary causative agent of urinary tract infections, typically express filamentous adhesive organelles called type 1 pili that mediate both bacterial attachment to and invasion of bladder urothelial cells. Several host proteins have previously been identified as receptors for type 1 pili, but none have been conclusively shown to promote UPEC entry into host bladder cells.

Danelle S. Eto; Tiffani A. Jones; Jamie L. Sundsbak; Matthew A. Mulvey

2007-01-01

11

Origins and Virulence Mechanisms of Uropathogenic Escherichia coli  

PubMed Central

Strains of uropathogenic E. coli (UPEC) are the primary cause of urinary tract infections, including both cystitis and pyelonephritis. These bacteria have evolved a multitude of virulence factors and strategies that facilitate bacterial growth and persistence within the adverse settings of the host urinary tract. Expression of adhesive organelles like type 1 and P pili allow UPEC to bind and invade host cells and tissues within the urinary tract while expression of iron chelating factors (siderophores) enable UPEC to pilfer host iron stores. Deployment of an array of toxins, including hemolysin and cytotoxic necrotizing factor 1, provide UPEC with the means to inflict extensive tissue damage, facilitating bacterial dissemination as well as releasing host nutrients and disabling immune effector cells. These toxins also have the capacity to modulate, in more subtle ways, host signaling pathways affecting myriad processes, including inflammatory responses, host cell survival, and cytoskeletal dynamics. Here, we discuss the mechanisms by which these and other virulence factors promote UPEC survival and growth within the urinary tract. Comparisons are also made between UPEC and other strains of extraintestinal pathogenic E. coli that, although closely related to UPEC, are distinct in their abilities to colonize the host and cause disease. PMID:18482721

Wiles, Travis J.; Kulesus, Richard R.; Mulvey, Matthew A.

2008-01-01

12

Escherichia coli–Mediated Impairment of Ureteric Contractility Is Uropathogenic E. coli Specific  

PubMed Central

Background.?Ureters are fundamental for keeping kidneys free from uropathogenic Escherichia coli (UPEC), but we have shown that 2 strains (J96 and 536) can subvert this role and reduce ureteric contractility. To determine whether this is (1) a widespread feature of UPEC, (2) exhibited only by UPEC, and (3) dependent upon type 1 fimbriae, we analyzed strains representing epidemiologically important multilocus sequence types ST131, ST73, and ST95 and non-UPEC E. coli. Methods.?Contractility and calcium transients in intact rat ureters were compared between strains. Mannose and fim mutants were used to investigate the role of type 1 fimbriae. Results.?Non-UPEC had no significant effect on contractility, with a mean decrease after 8 hours of 8.8%, compared with 8.8% in controls. UPEC effects on contractility were strain specific, with decreases from 9.47% to 96.7%. Mannose inhibited the effects of the most potent strains (CFT073 and UTI89) but had variable effects among other UPEC strains. Mutation and complementation studies showed that the effects of the UTI89 cystitis isolate were fimH dependent. Conclusions.?We find that (1) non-UPEC do not affect ureteric contractility, (2) impairment of contractility is a common feature of UPEC, and (3) the mechanism varies between strains, but for the most potent UPEC type 1 fimbriae are involved. PMID:23002447

Floyd, Rachel V.; Upton, Mathew; Hultgren, Scott J.; Wray, Susan; Burdyga, Theodor V.; Winstanley, Craig

2012-01-01

13

Prevalence and Persistence of Escherichia coli Strains with Uropathogenic Virulence Characteristics in Sewage Treatment Plants?  

PubMed Central

We investigated the prevalence and persistence of Escherichia coli strains in four sewage treatment plants (STPs) in a subtropical region of Queensland, Australia. In all, 264 E. coli strains were typed using a high-resolution biochemical fingerprinting method and grouped into either a single or a common biochemical phenotype (S-BPT and C-BPT, respectively). These strains were also tested for their phylogenetic groups and 12 virulence genes associated with intestinal and extraintestinal E. coli strains. Comparison of BPTs at various treatment stages indicated that certain BPTs were found in two or all treatment stages. These BPTs constituted the highest proportion of E. coli strains in each STP and belonged mainly to phylogenetic group B2 and, to a lesser extent, group D. No virulence genes associated with intestinal E. coli were found among the strains, but 157 (59.5%) strains belonging to 14 C-BPTs carried one or more virulence genes associated with uropathogenic strains. Of these, 120 (76.4%) strains belonged to seven persistent C-BPTs and were found in all four STPs. Our results indicate that certain clonal groups of E. coli with virulence characteristics of uropathogenic strains can survive the treatment processes of STPs. These strains were common to all STPs and constituted the highest proportion of the strains in different treatment tanks of each STP. PMID:20622128

Anastasi, E. M.; Matthews, B.; Gundogdu, A.; Vollmerhausen, T. L.; Ramos, N. L.; Stratton, H.; Ahmed, W.; Katouli, M.

2010-01-01

14

Oral Consumption of Cranberry Juice Cocktail Inhibits Molecular-Scale Adhesion of Clinical Uropathogenic Escherichia coli  

PubMed Central

Abstract Cranberry juice cocktail (CJC) has been shown to inhibit the formation of biofilm by uropathogenic Escherichia coli. In order to investigate whether the anti-adhesive components could reach the urinary tract after oral consumption of CJC, a volunteer was given 16?oz of either water or CJC. Urine samples were collected at 0, 2, 4, 6, and 8 hours after consumption of a single dose. The ability of compounds in the urine to influence bacterial adhesion was tested for six clinical uropathogenic E. coli strains, including four P-fimbriated strains (B37, CFT073, BF1023, and J96) and two strains not expressing P-fimbriae but exhibiting mannose-resistant hemagglutination (B73 and B78). A non-fimbriated strain, HB101, was used as a control. Atomic force microscopy (AFM) was used to measure the adhesion force between a silicon nitride probe and bacteria treated with urine samples. Within 2 hours after CJC consumption, bacteria of the clinical strains treated with the corresponding urine sample demonstrated lower adhesion forces than those treated with urine collected before CJC consumption. The adhesion forces continued decreasing with time after CJC consumption over the 8-hour measurement period. The adhesion forces of bacteria after exposure to urine collected following water consumption did not change. HB101 showed low adhesion forces following both water and CJC consumption, and these did not change over time. The AFM adhesion force measurements were consistent with the results of a hemagglutination assay, confirming that oral consumption of CJC could act against adhesion of uropathogenic E. coli. PMID:21480803

Tao, Yuanyuan; Pinzón-Arango, Paola A.; Howell, Amy B.

2011-01-01

15

Genome of multidrug-resistant uropathogenic Escherichia coli strain NA114 from India.  

PubMed

Uropathogenic Escherichia coli (UPEC) causes serious infections in people at risk and has a significant environmental prevalence due to contamination by human and animal excreta. In developing countries, UPEC assumes importance in certain dwellings because of poor community/personal hygiene and exposure to contaminated water or soil. We report the complete genome sequence of E. coli strain NA114 from India, a UPEC strain with a multidrug resistance phenotype and the capacity to produce extended-spectrum beta-lactamase. The genome sequence and comparative genomics emanating from it will be significant in under-standing the genetic makeup of diverse UPEC strains and in boosting the development of new diagnostics/vaccines. PMID:21685291

Avasthi, Tiruvayipati Suma; Kumar, Narender; Baddam, Ramani; Hussain, Arif; Nandanwar, Nishant; Jadhav, Savita; Ahmed, Niyaz

2011-08-01

16

Genome of Multidrug-Resistant Uropathogenic Escherichia coli Strain NA114 from India ?  

PubMed Central

Uropathogenic Escherichia coli (UPEC) causes serious infections in people at risk and has a significant environmental prevalence due to contamination by human and animal excreta. In developing countries, UPEC assumes importance in certain dwellings because of poor community/personal hygiene and exposure to contaminated water or soil. We report the complete genome sequence of E. coli strain NA114 from India, a UPEC strain with a multidrug resistance phenotype and the capacity to produce extended-spectrum beta-lactamase. The genome sequence and comparative genomics emanating from it will be significant in under-standing the genetic makeup of diverse UPEC strains and in boosting the development of new diagnostics/vaccines. PMID:21685291

Avasthi, Tiruvayipati Suma; Kumar, Narender; Baddam, Ramani; Hussain, Arif; Nandanwar, Nishant; Jadhav, Savita; Ahmed, Niyaz

2011-01-01

17

Draft genome sequences of five recent human uropathogenic Escherichia coli isolates  

PubMed Central

This study reports the release of draft genome sequences of five isolates of uropathogenic Escherichia coli (UPEC), isolated from patients suffering from uncomplicated cystitis in 2012 in Ann Arbor, Michigan. Phylogenetic analyses revealed that these strains belonged to E. coli phylogroups B2 and D, and are closely related to known UPEC strains. Comparative genomic analysis revealed that more conserved proteins were shared between these recent isolates and UPEC strains causing cystitis than those causing pyelonephritis. Additional genomic comparisons identified that three isolates encode a type III secretion system (T3SS) and a putative T3SS effector gene cluster along with an invasin-like outer membrane protein. Presence of T3SS genes is a rare occurrence among UPEC strains. These genomes further substantiate the heterogeneity of the gene pool of UPEC and provide a foundation for comparative genomic studies using recent clinical isolates. PMID:23821517

Subashchandrabose, Sargurunathan; Hazen, Tracy H.; Rasko, David A.; Mobley, Harry L. T.

2013-01-01

18

Urine post equivalent daily cranberry juice consumption may opsonize uropathogenicity of Escherichia coli.  

PubMed

Basic studies have proven that cranberries may prevent urinary tract infections through changing the adhesiveness of Escherichia coli (E. coli) to urothelial cells. Various cranberry preparations, including extract powder, capsules, and juice, have been shown to be effective in clinical and epidemiological research. Because cranberries are most commonly consumed as juice in a diluted concentration, the aim of this study was to investigate whether the equivalent daily dose of cranberry juice is sufficient to modify host urine to change the uropathogenicity of E. coli. Urine from rats taking an equivalent daily dose of cranberry juice has been shown to decrease the capability of E. coli in hemagglutination, urothelium adhesion, nematode killing, and biofilm formation. All these changes occurred after E. coli was incubated in cranberry metabolite-containing urine, defined as urine opsonization. Urine opsonization of E. coli resulted in 40.9% (p = 0.0038) decrease in hemagglutination ability, 66.7% (p = 0.0181) decrease in urothelium adhesiveness, 16.7% (p = 0.0004) increase in the 50% lethal time in killing nematodes, and 53.9% (p = 5.9 × 10(-4)) decrease in biofilm formation. Thus, an equivalent daily dose of cranberry juice should be considered sufficiently potent to demonstrate urine opsonization in E. coli. PMID:23440506

Chen, Chih-Shou; Ho, Dong-Ru; Chang, Pey-Jium; Lin, Wei-Yu; Huang, Yun-Ching

2013-10-01

19

Uropathogenic Escherichia coli Toxins Induce Caspase-Independent Apoptosis in Renal Proximal Tubular Cells via ERK Signaling  

Microsoft Academic Search

Background: Pyelonephritis is a risk factor for renal tubular epithelial cell damage. Recent studies have shown that Escherichia coli and\\/or its toxins may stimulate apoptotic cell death in renal tubular cells, but the underlying molecular mechanisms remain to be elucidated. Methods: Confluent LLC-PK1 cells were exposed to E. coli toxins from overnight cultures of the uropathogenic O6K13H1 (O6) and the

Ming Chen; Timo Jahnukainen; Wenjie Bao; Elisabetta Daré; Sandra Ceccatelli; Gianni Celsi

2003-01-01

20

Transcriptional Responses of Uropathogenic Escherichia coli to Increased Environmental Osmolality Caused by Salt or Urea  

PubMed Central

Uropathogenic Escherichia coli (UPEC) is the most common causative agent of urinary tract infections in humans. The majority of urinary infections develop via ascending route through the urethra, where bacterial cells come in contact with human urine prior to reaching the bladder or kidneys. Since urine contains significant amounts of inorganic ions and urea, it imposes osmotic and denaturing stresses on bacterial cells. In this study, we determined the transcriptional adaptive responses of UPEC strain CFT073 to the presence of 0.3 M NaCl or 0.6 M urea in the growth medium. The cell responses to these two osmolytes were drastically different. Although most of the genes of the osmotically inducible regulon were overexpressed in medium with salt, urea failed to stimulate osmotic stress response. At the same time, UPEC colonization genes encoding type 1 and F1C fimbriae and capsule biosynthesis were transcriptionally induced in the presence of urea but did not respond to increased salt concentration. We speculate that urea can potentially be sensed by uropathogenic bacteria to initiate infection program. In addition, several molecular chaperone genes were overexpressed in the presence of urea, whereas adding NaCl to the medium led to an upregulation of a number of anaerobic metabolism pathways. PMID:23090957

Withman, Benjamin; Gunasekera, Thusitha S.; Beesetty, Pavani; Agans, Richard

2013-01-01

21

Hemagglutination ability and adherence to the Buffalo green monkey kidney cell line of uropathogenic Escherichia coli.  

PubMed

The hemagglutination ability and adherence capacity to the Buffalo green monkey (BGM) kidney cell line of 160 wild-type strains of Escherichia coli isolated from bacteriuric patients were investigated. It was found that P-fimbriated E. coli strains adhered significantly better to BGM cells than did strains in which P-fimbriae were not detected, which is in accordance with the capacity of P-fimbriated strains to cause unobstructive pyelonephritis and with receptor distribution for P-fimbriae in the urinary tract. The strains which exhibited other adhesions, alone or simultaneously, showed reduced adherence to BGM cells, while non-agglutinating strains, mostly isolated from urine of patients with asymptomatic bacteriuria, did not adhere at all or adhered poorly to the utilized cell line. The BGM cells served as a good experimental model for investigation of uropathogenic E. coli adherence; because these cells originate from the upper urinary tract, they are viable and not coated with Tamm-Horsfall protein. PMID:9393553

Vranes, J

1997-11-01

22

Medicinal plants extracts affect virulence factors expression and biofilm formation by the uropathogenic Escherichia coli.  

PubMed

Medicinal plants are an important source for the therapeutic remedies of various diseases including urinary tract infections. This prompted us to perform research in this area. We decided to focus on medicinal plants species used in urinary tract infections prevention. The aim of our study was to determine the influence of Betula pendula, Equisetum arvense, Herniaria glabra, Galium odoratum, Urtica dioica, and Vaccinium vitis-idaea extracts on bacterial survival and virulence factors involved in tissue colonization and biofilm formation of the uropathogenic Escherichia coli rods. Qualitative and quantitative analysis of plant extracts were performed. Antimicrobial assay relied on the estimation of the colony forming unit number. Hydrophobicity of cells was established by salt aggregation test. Using motility agar, the ability of bacteria to move was examined. The erythrocyte hemagglutination test was used for fimbriae P screening. Curli expression was determined using YESCA agar supplemented with congo red. Quantification of biofilm formation was carried out using a microtiter plate assay and a spectrophotometric method. The results of the study indicate significant differences between investigated extracts in their antimicrobial activities. The extracts of H. glabra and V. vitis-idaea showed the highest growth-inhibitory effects (p < 0.05). Surface hydrophobicity of autoaggregating E. coli strain changed after exposure to all plant extracts, except V. vitis-idaea (p > 0.05). The B. pendula and U. dioica extracts significantly reduced the motility of the E. coli rods (p < 0.05). All the extracts exhibited the anti-biofilm activity. PMID:22915095

Wojnicz, Dorota; Kucharska, Alicja Z; Sokó?-??towska, Anna; Kicia, Marta; Tichaczek-Goska, Dorota

2012-12-01

23

Tetracycline rapidly reaches all the constituent cells of uropathogenic Escherichia coli biofilms.  

PubMed

We have developed a method for visualizing Escherichia coli cells that are exposed to tetracycline in a biofilm, based on a previous report that liposomes containing the E. coli TetR(B) protein fluoresce when exposed to this antibiotic. By our method, cells devoid of TetR(B) also exhibited tetracycline-dependent fluorescence. At 50 microg of tetracycline ml(-1), planktonic cells of a uropathogenic E. coli (UPEC) strain developed maximal fluorescence after 7.5 to 10 min of exposure. A similar behavior was exhibited by cells in a 24- or 48-h UPEC biofilm, as examined by confocal laser microscopy, regardless of whether they lined empty spaces or occupied densely packed regions. Further, a comparison of phase-contrast and fluorescent images of corresponding biofilm zones showed that all the cells fluoresced. Thus, all the biofilm cells were exposed to tetracycline and there were no pockets within the biofilm where the antibiotic failed to reach. It also appeared unlikely that niches of reduced exposure to the antibiotic existed within the biofilms. PMID:12121918

Stone, G; Wood, P; Dixon, L; Keyhan, M; Matin, A

2002-08-01

24

Virulence factors and genetic variability of uropathogenic Escherichia coli isolated from dogs and cats in Italy.  

PubMed

In this study, the association between virulence genotypes and phylogenetic groups among Escherichia (E.) coli isolates obtained from pet dogs and cats with cystitis was detected, and fingerprinting methods were used to explore the relationship among strains. Forty uropathogenic E. coli (UPEC) isolated from dogs (n = 30) and cats (n = 10) in Italy were analysed by polymerase chain reaction (PCR) for the presence of virulence factors and their classification into phylogenetic groups. The same strains were characterized by repetitive extragenic palindromic (REP)- and enterobacterial repetitive intergenic consensus (ERIC)-PCR techniques. We found a high number of virulence factors such as fimbriae A, S fimbriae (sfa) and cytotoxic necrotizing factor 1 (cnf1) significantly associated with phylogenetic group B2. We demonstrated a high correlation between ?-hemolysin A and pyelonephritis C, sfa, and cnf1 operons, confirming the presence of pathogenicity islands in these strains. In addition, UPEC belonging to group B2 harboured a greater number of virulence factors than strains from phylogenetic groups A, B1, and D. REP- and ERIC-PCR grouped the UPEC isolates into two major clusters, the former grouping E. coli strains belonging to phylogenetic group B2 and D, the latter grouping those belonging to groups A and B1. Given the significant genetic variability among the UPEC strains found in our study, it can be hypothesized that no specific genotype is responsible for cystitis in cats or dogs. PMID:21368563

Tramuta, Clara; Nucera, Daniele; Robino, Patrizia; Salvarani, Sara; Nebbia, Patrizia

2011-03-01

25

Virulence factors and genetic variability of uropathogenic Escherichia coli isolated from dogs and cats in Italy  

PubMed Central

In this study, the association between virulence genotypes and phylogenetic groups among Escherichia (E.) coli isolates obtained from pet dogs and cats with cystitis was detected, and fingerprinting methods were used to explore the relationship among strains. Forty uropathogenic E. coli (UPEC) isolated from dogs (n = 30) and cats (n = 10) in Italy were analysed by polymerase chain reaction (PCR) for the presence of virulence factors and their classification into phylogenetic groups. The same strains were characterized by repetitive extragenic palindromic (REP)- and enterobacterial repetitive intergenic consensus (ERIC)-PCR techniques. We found a high number of virulence factors such as fimbriae A, S fimbriae (sfa) and cytotoxic necrotizing factor 1 (cnf1) significantly associated with phylogenetic group B2. We demonstrated a high correlation between ?-hemolysin A and pyelonephritis C, sfa, and cnf1 operons, confirming the presence of pathogenicity islands in these strains. In addition, UPEC belonging to group B2 harboured a greater number of virulence factors than strains from phylogenetic groups A, B1, and D. REP- and ERIC-PCR grouped the UPEC isolates into two major clusters, the former grouping E. coli strains belonging to phylogenetic group B2 and D, the latter grouping those belonging to groups A and B1. Given the significant genetic variability among the UPEC strains found in our study, it can be hypothesized that no specific genotype is responsible for cystitis in cats or dogs. PMID:21368563

Tramuta, Clara; Nucera, Daniele; Robino, Patrizia; Salvarani, Sara

2011-01-01

26

The structure of cranberry proanthocyanidins which inhibit adherence of uropathogenic P-fimbriated Escherichia coli in vitro  

Microsoft Academic Search

Ethyl acetate extracts of Sephadex LH20-purified proanthocyanidins of American cranberry (Vaccinium macrocarpon Ait.) exhibited potent biological activity by inhibiting adherence of uropathogenic isolates of P-fimbriated Escherichia coli bacteria to cellular surfaces containing ?-Gal(1 ? 4)?-Gal receptor sequences similar to those on epithelial cells in the urinary tract. The chemical structures of the proanthocyanidins were determined by 13C NMR, electrospray mass

Lai Yeap Foo; Yinrong Lu; Amy B Howell; Nicholi Vorsa

2000-01-01

27

Biofilm formation and virulence of uropathogenic Escherichia coli in urine after consumption of cranberry-lingonberry juice  

Microsoft Academic Search

Cranberry-lingonberry juice (CLJ) was effective in preventing urinary tract infections (UTIs) in our earlier randomized clinical\\u000a trial. We aimed to test whether consumption of CLJ at a similar dose to earlier reduces the biofilm formation and virulence\\u000a of uropathogenic Escherichia coli in urine. Twenty healthy women drank 100 ml of CLJ daily for two weeks. Urine samples were obtained 2–4 hours

T. Tapiainen; H. Jauhiainen; L. Jaakola; J. Salo; J. Sevander; I. Ikäheimo; A. M. Pirttilä; A. Hohtola; M. Uhari

28

Drug resistance and virulence of uropathogenic Escherichia coli from Shanghai, China.  

PubMed

Uropathogenic Escherichia coli (UPEC) is the major cause of urinary tract infections (UTIs). In the present study, 198 E. coli isolates from patients with UTIs in Shanghai in 2008 were examined by susceptibility testing, with an extremely high number (153/198) showing multidrug resistance (MDR). And, the expression of extended-spectrum ?-lactamases (ESBLs) reached 48.5% (96/198). The resistance rates to penicillins, fluoroquinolone, folate pathway inhibitors and first- and second-generation cephalosporins were high. Molecular analyses showed that the CTX-M-9 group (70/96) was the most common CTX-M group among UPEC, followed by the CTX-M-1 group (27/96). Phylogenetic group D accounted for 42.4% (84/198) of the isolates, exhibiting the highest ESBLs (50/84) and MDR (75/84) rates. Virulence genes were present in a significantly high proportion in the phylogenetic group B2 isolates, except for the afaBC gene. The ESBL-producing strains analyzed by pulsed-field gel electrophoresis (PFGE) were clustered into six groups at a cutoff of 67%. Notably, the findings that afaBC was specific to phylogenetic group D and PFGE group I and was correlated with the CTX-M-9 group were different from a previous report. In conclusion, knowledge of antimicrobial resistance data and virulence factors may enable clinicians to tailor empirical antibiotic treatments for UTIs. PMID:24984795

Wang, Yanchun; Zhao, Shengyuan; Han, Lizhong; Guo, Xiaokui; Chen, Min; Ni, Yuxing; Zhang, Yan; Cui, Zelin; He, Ping

2014-12-01

29

Alkaloids modulate motility, biofilm formation and antibiotic susceptibility of uropathogenic Escherichia coli.  

PubMed

Alkaloid-containing natural compounds have shown promise in the treatment of microbial infections. However, practical application of many of these compounds is pending a mechanistic understanding of their mode of action. We investigated the effect of two alkaloids, piperine (found in black pepper) and reserpine (found in Indian snakeroot), on the ability of the uropathogenic bacterium Escherichia coli CFT073 to colonize abiotic surfaces. Sub-inhibitory concentrations of both compounds (0.5 to 10 µg/mL) decreased bacterial swarming and swimming motilities and increased biofilm formation. qRT-PCR revealed a decrease in the expression of the flagellar gene (fliC) and motility genes (motA and motB) along with an increased expression of adhesin genes (fimA, papA, uvrY). Interestingly, piperine increased penetration of the antibiotics ciprofloxacin and azithromycin into E. coli CFT073 biofilms and consequently enhanced the ability of these antibiotics to disperse pre-established biofilms. The findings suggest that these alkaloids can potentially affect bacterial colonization by hampering bacterial motility and may aid in the treatment of infection by increasing antibiotic penetration in biofilms. PMID:25391152

Dusane, Devendra H; Hosseinidoust, Zeinab; Asadishad, Bahareh; Tufenkji, Nathalie

2014-01-01

30

Alkaloids Modulate Motility, Biofilm Formation and Antibiotic Susceptibility of Uropathogenic Escherichia coli  

PubMed Central

Alkaloid-containing natural compounds have shown promise in the treatment of microbial infections. However, practical application of many of these compounds is pending a mechanistic understanding of their mode of action. We investigated the effect of two alkaloids, piperine (found in black pepper) and reserpine (found in Indian snakeroot), on the ability of the uropathogenic bacterium Escherichia coli CFT073 to colonize abiotic surfaces. Sub-inhibitory concentrations of both compounds (0.5 to 10 µg/mL) decreased bacterial swarming and swimming motilities and increased biofilm formation. qRT-PCR revealed a decrease in the expression of the flagellar gene (fliC) and motility genes (motA and motB) along with an increased expression of adhesin genes (fimA, papA, uvrY). Interestingly, piperine increased penetration of the antibiotics ciprofloxacin and azithromycin into E. coli CFT073 biofilms and consequently enhanced the ability of these antibiotics to disperse pre-established biofilms. The findings suggest that these alkaloids can potentially affect bacterial colonization by hampering bacterial motility and may aid in the treatment of infection by increasing antibiotic penetration in biofilms. PMID:25391152

Dusane, Devendra H.; Hosseinidoust, Zeinab; Asadishad, Bahareh; Tufenkji, Nathalie

2014-01-01

31

How to become a uropathogen: Comparative genomic analysis of extraintestinal pathogenic Escherichia coli strains  

PubMed Central

Uropathogenic Escherichia coli (UPEC) strain 536 (O6:K15:H31) is one of the model organisms of extraintestinal pathogenic E. coli (ExPEC). To analyze this strain's genetic basis of urovirulence, we sequenced the entire genome and compared the data with the genome sequence of UPEC strain CFT073 (O6:K2:H1) and to the available genomes of nonpathogenic E. coli strain MG1655 (K-12) and enterohemorrhagic E. coli. The genome of strain 536 is ?292 kb smaller than that of strain CFT073. Genomic differences between both UPEC are mainly restricted to large pathogenicity islands, parts of which are unique to strain 536 or CFT073. Genome comparison underlines that repeated insertions and deletions in certain parts of the genome contribute to genome evolution. Furthermore, 427 and 432 genes are only present in strain 536 or in both UPEC, respectively. The majority of the latter genes is encoded within smaller horizontally acquired DNA regions scattered all over the genome. Several of these genes are involved in increasing the pathogens' fitness and adaptability. Analysis of virulence-associated traits expressed in the two UPEC O6 strains, together with genome comparison, demonstrate the marked genetic and phenotypic variability among UPEC. The ability to accumulate and express a variety of virulence-associated genes distinguishes ExPEC from many commensals and forms the basis for the individual virulence potential of ExPEC. Accordingly, instead of a common virulence mechanism, different ways exist among ExPEC to cause disease. PMID:16912116

Brzuszkiewicz, Elzbieta; Brüggemann, Holger; Liesegang, Heiko; Emmerth, Melanie; Ölschläger, Tobias; Nagy, Gábor; Albermann, Kaj; Wagner, Christian; Buchrieser, Carmen; Em?dy, Levente; Gottschalk, Gerhard; Hacker, Jörg; Dobrindt, Ulrich

2006-01-01

32

Virulence Factors and O-Serogroups Profiles of Uropathogenic Escherichia Coli Isolated from Iranian Pediatric Patients  

PubMed Central

Background: Uropathogenic Escherichia coli O- Serogroups with their virulence factors are the most prevalent causes of UTIs. Objectives: The present investigation was performed to study the virulence factors and O-Serogroups profiles of UPEC isolated from Iranian pediatric patients. Patients and Methods: This cross sectional investigation was performed on 100 urine samples collected from hospitalized pediatrics of Baqiyatallah Hospital, Tehran, Iran. Midstream urine was collected to decrease potential bacterial, cellular and artifactual contamination. All samples were cultured and those with positive results were subjected to polymerase chain reactions to detect pap, cnf1, afa, sfa and hlyA genes and various O- Serogroups. Results: We found that 37.5% of boys and 75% of girls had positive results for Escherichia coli. We also found that O1 (19.33%), O2 (13.33%), O6 (13.33%), O4 (11.66%), and O18 (11.66 %) were the most commonly detected Serogroups. Totally, the serogroup of 5% of all strains were not detected. In addition, all of these O- Serogroups were pap+, cnf1+, hlyA+, and afa+. Totally, pap (70 %), cnf1 (56.66 %), and hlyA (43.33 %) were the most commonly detected virulence genes in the both studied groups of children. The sfa (30 %) and afa (26.66 %) genes had the lowest incidence rates. Conclusions: Special health care should be performed on UTIs management in Iranian pediatric patients. Extended researches should be performed to evaluate relation between other O-Serogroups and virulent genes. PMID:24719745

Dormanesh, Banafshe; Safarpoor Dehkordi, Farhad; Hosseini, Sahar; Momtaz, Hassan; Mirnejad, Reza; Hoseini, Mohammad Javad; Yahaghi, Emad; Tarhriz, Vahideh; Khodaverdi Darian, Ebrahim

2014-01-01

33

Uropathogenic Escherichia coli in Iran: Serogroup distributions, virulence factors and antimicrobial resistance properties  

PubMed Central

Background Urinary tract infections (UTIs) are one of the most common bacterial infections with global expansion. These infections are predominantly caused by uropathogenic Escherichia coli (UPEC). Methods Totally, 123 strains of Escherichia coli isolated from UTIs patients, using bacterial culture method were subjected to polymerase chain reactions for detection of various O- serogroups, some urovirulence factors, antibiotic resistance genes and resistance to 13 different antibiotics. Results According to data, the distribution of O1, O2, O6, O7 and O16 serogroups were 2.43%, besides O22, O75 and O83 serogroups were 1.62%. Furthermore, the distribution of O4, O8, O15, O21 and O25 serogroups were 5.69%, 3.25%, 21.13%, 4.06% and 26.01%, respectively. Overall, the fim virulence gene had the highest (86.17%) while the usp virulence gene had the lowest distributions of virulence genes in UPEC strains isolated from UTIs patients. The vat and sen virulence genes were not detected in any UPEC strains. Totally, aadA1 (52.84%), and qnr (46.34%) were the most prevalent antibiotic resistance genes while the distribution of cat1 (15.44%), cmlA (15.44%) and dfrA1 (21.95%) were the least. Resistance to penicillin (100%) and tetracycline (73.98%) had the highest while resistance to nitrofurantoin (5.69%) and trimethoprim (16.26%) had the lowest frequencies. Conclusions This study indicated that the UPEC strains which harbored the high numbers of virulence and antibiotic resistance genes had the high ability to cause diseases that are resistant to most antibiotics. In the current situation, it seems that the administration of penicillin and tetracycline for the treatment of UTIs is vain. PMID:23627669

2013-01-01

34

Pleiotropic Roles of uvrY on Biofilm Formation, Motility and Virulence in Uropathogenic Escherichia coli CFT073  

PubMed Central

Urinary tract infections primarily caused by uropathogenic strains of Escherichia coli (E. coli) remain a significant public health problem in both developed and developing countries. An important virulence determinant in uropathogenesis is biofilm formation which requires expression of fimbriae, flagella, and other surface components such as lipopolysaccharides. In this study, we explored the regulation of uvrY and csrA genes in biofilm formation, motility and virulence determinants in uropathogenic E. coli. We found that mutation in uvrY suppressed biofilm formation on abiotic surfaces such as polyvinyl chloride, polystyrene and glass, and complementation of uvrY in the mutant restored the biofilm phenotype. We further evaluated the role of uvrY gene in expression of type 1 fimbriae, an important adhesin that facilitates adhesion to various abiotic surfaces. We found that phase variation of type 1 fimbriae between fimbriated and afimbriated mode was modulated by uvrY at its transcriptional level. Deletion mutant of uvrY lowered expression of fimbrial recombinase genes, such as fimB, fimE, and fimA, a gene encoding major fimbrial subunit. Furthermore, transcription of virulence specific genes such as papA, hlyB and galU was also reduced in the deletion mutant. Swarming motility and expression of flhD and flhC was also diminished in the mutant. Taken together, our findings unravel a possible mechanism in which uvrY facilitates biofilm formation, persistence and virulence of uropathogenic E. coli. PMID:23383333

Mitra, Arindam; Palaniyandi, Senthilkumar; Herren, Christopher D.; Zhu, Xiaoping; Mukhopadhyay, Suman

2013-01-01

35

Comparison of adhesin genes and antimicrobial susceptibilities between uropathogenic and intestinal commensal Escherichia coli strains.  

PubMed

The presence of adhesins is arguably an important determinant of pathogenicity for Uropathogenic Escherichia coli (UPEC). Antimicrobial susceptibilities were tested by agar dilution method, fifteen adhesin genes were detected by polymerase chain reaction, and multilocus sequence typing (MLST) was analyzed in 70 UPEC isolates and 41 commensal E. coli strains. Extended-spectrum ?-lactamase (ESBL) was determined with confirmatory test. The prevalence of ESBL-producers in UPEC (53%, 37/70) was higher than the commensal intestinal isolates (7%, 3/41), and 97% (36/37) of the ESBL-producing UPEC harbored bla CTX-M genes. afa was present in 36% (10/28) UPEC isolates from recurrent lower urinary tract infection (UTI), and none in the acute pyelonephritis, acute uncomplicated cystitis or commensal strains (P<0.0001). papG was detected in 28% (20/70) of UPEC isolates, while 5% (2/41) of the commensal strains were papG positive (P?=?0.0025), and the prevalence of papG was significantly higher in acute pyelonephritis group (71%) than the other two UTI groups (P<0.0001). The prevalence of flu, yqi, yadN and ygiL was significantly higher in UPEC isolates than in the commensal strains. ESBL-producing UPEC showed a lower prevalence of adhesin genes compared with non-ESBL-producing strains. The MLST profiles were different between UPEC and commensal strains, with ST131 (19%, 13/70) and ST10 (20%, 8/41) being the most common MLSTs, respectively. This study demonstrated that several adhesin genes were more prevalent in UPEC isolates than in commensal E. coli, and afa may be associated with recurrent lower UTI whereas papG is more frequently associated with acute pyelonephritis. PMID:23593422

Qin, Xiaohua; Hu, Fupin; Wu, Shi; Ye, Xinyu; Zhu, Demei; Zhang, Ying; Wang, Minggui

2013-01-01

36

Waging War against Uropathogenic Escherichia coli: Winning Back the Urinary Tract?  

PubMed Central

Urinary tract infection (UTI) caused by uropathogenic Escherichia coli (UPEC) is a substantial economic and societal burden—a formidable public health issue. Symptomatic UTI causes significant discomfort in infected patients, results in lost productivity, predisposes individuals to more serious infections, and usually necessitates antibiotic therapy. There is no licensed vaccine available for prevention of UTI in humans in the United States, likely due to the challenge of targeting a relatively heterogeneous group of pathogenic strains in a unique physiological niche. Despite significant advances in the understanding of UPEC biology, mechanistic details regarding the host response to UTI and full comprehension of genetic loci that influence susceptibility require additional work. Currently, there is an appreciation for the role of classic innate immune responses—from pattern receptor recognition to recruitment of phagocytic cells—that occur during UPEC-mediated UTI. There is, however, a clear disconnect regarding how factors involved in the innate immune response to UPEC stimulate acquired immunity that facilitates enhanced clearance upon reinfection. Unraveling the molecular details of this process is vital in the development of a successful vaccine for prevention of human UTI. Here, we survey the current understanding of host responses to UPEC-mediated UTI with an eye on molecular and cellular factors whose activity may be harnessed by a vaccine that stimulates lasting and sterilizing immunity. PMID:19917708

Sivick, Kelsey E.; Mobley, Harry L. T.

2010-01-01

37

Role of Capsule and O Antigen in the Virulence of Uropathogenic Escherichia coli  

PubMed Central

Urinary tract infection (UTI) is one of the most common bacterial infections in humans, with uropathogenic Escherichia coli (UPEC) the leading causative organism. UPEC has a number of virulence factors that enable it to overcome host defenses within the urinary tract and establish infection. The O antigen and the capsular polysaccharide are two such factors that provide a survival advantage to UPEC. Here we describe the application of the rpsL counter selection system to construct capsule (kpsD) and O antigen (waaL) mutants and complemented derivatives of three reference UPEC strains: CFT073 (O6:K2:H1), RS218 (O18:K1:H7) and 1177 (O1:K1:H7). We observed that while the O1, O6 and O18 antigens were required for survival in human serum, the role of the capsule was less clear and linked to O antigen type. In contrast, both the K1 and K2 capsular antigens provided a survival advantage to UPEC in whole blood. In the mouse urinary tract, mutation of the O6 antigen significantly attenuated CFT073 bladder colonization. Overall, this study contrasts the role of capsule and O antigen in three common UPEC serotypes using defined mutant and complemented strains. The combined mutagenesis-complementation strategy can be applied to study other virulence factors with complex functions both in vitro and in vivo. PMID:24722484

Sarkar, Sohinee; Ulett, Glen C.; Totsika, Makrina; Phan, Minh-Duy; Schembri, Mark A.

2014-01-01

38

Transcriptional activation of a pap pilus virulence operon from uropathogenic Escherichia coli.  

PubMed Central

A gene cluster mediating production of pili in uropathogenic Escherichia coli was analysed with respect to regulation of pili synthesis. Two cistrons, papB and papI, were localized upstream of the major pilus subunit gene, papA. The papI-papB-papA region was characterized by nucleotide sequencing and by transcriptional analysis. The papA gene was primarily represented by an 800 nucleotide long transcript but was also co-transcribed with papB as a less abundant 1300 nucleotide long mRNA. Both transcripts presumably terminated at the same site downstream of the papA coding sequence. The weakly expressed papI gene was transcribed in the opposite direction to that of papB and papA. Studies with lacZ operon fusions showed that the papB gene encoded a trans-active effector required for papA transcription. Similarly, the papI gene stimulated papB transcription in trans. Furthermore, full expression of papA was cis dependent upon the papI-papB region. Transcription of the papB gene was shown to be dependent upon cAMP and its receptor protein. A binding site for the cAMP-CRP complex was postulated in the DNA sequence upstream of the papB promoter. Images Fig. 3. PMID:2868893

Båga, M; Göransson, M; Normark, S; Uhlin, B E

1985-01-01

39

Phenotypic Heterogeneity Enables Uropathogenic Escherichia coli To Evade Killing by Antibiotics and Serum Complement.  

PubMed

Uropathogenic strains of Escherichia coli (UPEC) are the major cause of bacteremic urinary tract infections. Survival in the bloodstream is associated with different mechanisms that help to resist serum complement-mediated killing. While the phenotypic heterogeneity of bacteria has been shown to influence antibiotic tolerance, the possibility that it makes cells refractory to killing by the immune system has not been experimentally tested. In the present study we sought to determine whether the heterogeneity of bacterial cultures is relevant to bacterial targeting by the serum complement system. We monitored cell divisions in the UPEC strain CFT073 with fluorescent reporter protein. Stationary-phase cells were incubated in active or heat-inactivated human serum in the presence or absence of different antibiotics (ampicillin, norfloxacin, and amikacin), and cell division and complement protein C3 binding were measured by flow cytometry and immunofluorescence microscopy. Heterogeneity in the doubling times of CFT073 cells in serum enabled three phenotypically different subpopulations to be distinguished, all of them being recognized by the C3 component of the complement system. The population of rapidly growing cells resists serum complement-mediated lysis. The dominant subpopulation of cells with intermediate growth rate is susceptible to serum. The third population, which does not resume growth upon dilution from stationary phase, is simultaneously protected from serum complement and antibiotics. PMID:25561706

Putrinš, Marta; Kogermann, Karin; Lukk, Eliisa; Lippus, Markus; Varik, Vallo; Tenson, Tanel

2015-03-01

40

Cytotoxic Necrotizing Factor Type 1 Production by Uropathogenic Escherichia coli Modulates Polymorphonuclear Leukocyte Function  

PubMed Central

Many strains of uropathogenic Escherichia coli (UPEC) produce cytotoxic necrotizing factor type 1 (CNF1), a toxin that constitutively activates the Rho GTPases RhoA, Rac1, and Cdc42. We previously showed that CNF1 contributes to the virulence of UPEC in a mouse model of ascending urinary tract infection and a rat model of acute prostatitis and that a striking feature of the histopathology of the mouse bladders and rat prostates infected with CNF1-positive strains is an elevation in levels of polymorphonuclear leukocytes (PMNs). We also found that CNF1 synthesis leads to prolonged survival of UPEC in association with human neutrophils. Here, we tested the hypothesis that CNF1 production by UPEC diminishes the antimicrobial capacity of mouse PMNs by affecting phagocyte function through targeting Rho family GTPases that are critical to phagocytosis and the generation of reactive oxygen species. We found that, as with human neutrophils, CNF1 synthesis provided a survival advantage to UPEC incubated with mouse PMNs. We also observed that CNF1-positive UPEC down-regulated phagocytosis, altered the distribution of the complement receptor CR3 (CD11b/CD18), enhanced the intracellular respiratory burst, and increased levels of Rac2 activation in PMNs. From these results, we conclude that modulation of PMN function by CNF1 facilitates UPEC survival during the acute inflammatory response. PMID:16113245

Davis, Jon M.; Rasmussen, Susan B.; O'Brien, Alison D.

2005-01-01

41

The Cpx Stress Response System Potentiates the Fitness and Virulence of Uropathogenic Escherichia coli  

PubMed Central

Strains of uropathogenic Escherichia coli (UPEC) are the primary cause of urinary tract infections, representing one of the most widespread and successful groups of pathogens on the planet. To colonize and persist within the urinary tract, UPEC must be able to sense and respond appropriately to environmental stresses, many of which can compromise the bacterial envelope. The Cpx two-component envelope stress response system is comprised of the inner membrane histidine kinase CpxA, the cytosolic response regulator CpxR, and the periplasmic auxiliary factor CpxP. Here, by using deletion mutants along with mouse and zebrafish infection models, we show that the Cpx system is critical to the fitness and virulence of two reference UPEC strains, the cystitis isolate UTI89 and the urosepsis isolate CFT073. Specifically, deletion of the cpxRA operon impaired the ability of UTI89 to colonize the murine bladder and greatly reduced the virulence of CFT073 during both systemic and localized infections within zebrafish embryos. These defects coincided with diminished host cell invasion by UTI89 and increased sensitivity of both strains to complement-mediated killing and the aminoglycoside antibiotic amikacin. Results obtained with the cpxP deletion mutants were more complicated, indicating variable strain-dependent and niche-specific requirements for this well-conserved auxiliary factor. PMID:23429541

Debnath, Irina; Norton, J. Paul; Barber, Amelia E.; Ott, Elizabeth M.; Dhakal, Bijaya K.; Kulesus, Richard R.

2013-01-01

42

High metabolic potential may contribute to the success of ST131 uropathogenic Escherichia coli.  

PubMed

Uropathogenic Escherichia coli (UPEC) is the predominant cause of urinary tract infection in both hospital and community settings. The recent emergence of multidrug-resistant clones like the O25b:H4-ST131 lineage represents a significant threat to health, and numerous studies have explored the virulence potential of these organisms. Members of the ST131 clone have been described as having variable carriage of key virulence factors, and it has been suggested that additional unidentified factors contribute to virulence. Here we demonstrated that ST131 isolates have high metabolic potential and biochemical profiles that distinguish them from isolates of many other sequence types (STs). A collection of 300 UPEC isolates recovered in 2007 and 2009 in the Northwest region of England were subjected to metabolic profiling using the Vitek2 Advanced Expert System (AES). Of the 47 tests carried out, 30 gave a positive result with at least one of the 300 isolates examined. ST131 isolates demonstrated significant association with eight tests, including those for peptidase, decarboxylase, and alkalinization activity. Metabolic activity also correlated with antibiotic susceptibility profiles, with resistant organisms displaying the highest metabolic potential. This is the first comprehensive study of metabolic potential in the ST131 lineage, and we suggest that high metabolic potential may have contributed to the fitness of members of the ST131 clone, which are able to exploit the available nutrients in both the intestinal and urinary tract environments. PMID:22814460

Gibreel, Tarek M; Dodgson, Andrew R; Cheesbrough, John; Bolton, Frederick J; Fox, Andrew J; Upton, Mathew

2012-10-01

43

Persistence of Uropathogenic Escherichia coli in the Face of Multiple Antibiotics ?  

PubMed Central

Numerous antibiotics have proven to be effective at ameliorating the clinical symptoms of urinary tract infections (UTIs), but recurrent and chronic infections continue to plague many individuals. Most UTIs are caused by strains of uropathogenic Escherichia coli (UPEC), which can form both extra- and intracellular biofilm-like communities within the bladder. UPEC also persist inside host urothelial cells in a more quiescent state, sequestered within late endosomal compartments. Here, we tested a panel of 17 different antibiotics, representing seven distinct functional classes, for their effects on the survival of the reference UPEC isolate UTI89 within both biofilms and host bladder urothelial cells. All but one of the tested antibiotics prevented UTI89 growth in broth culture, and most were at least modestly effective against bacteria present within in vitro-grown biofilms. In contrast, only a few of the antibiotics, including nitrofurantoin and the fluoroquinolones ciprofloxacin and sparfloxacin, were able to eliminate intracellular bacteria in bladder cell culture-based assays. However, in a mouse UTI model system in which these antibiotics reached concentrations in the urine specimens that far exceeded minimal inhibitory doses, UPEC reservoirs in bladder tissues were not effectively eradicated. We conclude that the persistence of UPEC within the bladder, regardless of antibiotic treatments, is likely facilitated by a combination of biofilm formation, entry of UPEC into a quiescent or semiquiescent state within host cells, and the stalwart permeability barrier function associated with the bladder urothelium. PMID:20231390

Blango, Matthew G.; Mulvey, Matthew A.

2010-01-01

44

Characterization of a Dipartite Iron Uptake System from Uropathogenic Escherichia coli Strain F11*  

PubMed Central

In the uropathogenic Escherichia coli strain F11, in silico genome analysis revealed the dicistronic iron uptake operon fetMP, which is under iron-regulated control mediated by the Fur regulator. The expression of fetMP in a mutant strain lacking known iron uptake systems improved growth under iron depletion and increased cellular iron accumulation. FetM is a member of the iron/lead transporter superfamily and is essential for iron uptake by the Fet system. FetP is a periplasmic protein that enhanced iron uptake by FetM. Recombinant FetP bound Cu(II) and the iron analog Mn(II) at distinct sites. The crystal structure of the FetP dimer reveals a copper site in each FetP subunit that adopts two conformations: CuA with a tetrahedral geometry composed of His44, Met90, His97, and His127, and CuB, a second degenerate octahedral geometry with the addition of Glu46. The copper ions of each site occupy distinct positions and are separated by ?1.3 ?. Nearby, a putative additional Cu(I) binding site is proposed as an electron source that may function with CuA/CuB displacement to reduce Fe(III) for transport by FetM. Together, these data indicate that FetMP is an additional iron uptake system composed of a putative iron permease and an iron-scavenging and potentially iron-reducing periplasmic protein. PMID:21596746

Koch, Doreen; Chan, Anson C. K.; Murphy, Michael E. P.; Lilie, Hauke; Grass, Gregor; Nies, Dietrich H.

2011-01-01

45

Integrons in uropathogenic Escherichia coli and their relationship with phylogeny and virulence.  

PubMed

Uropathogenic Escherichia coli (UPEC) comprise a heterogeneous group of strains. In a previous epidemiological survey performed on 230 UPEC isolates, five virulence profiles were described, each one defined by the presence of some virulence determinants and by the absence of others. Phylogenetic groups and antibiotic resistances distributed non-randomly among the isolates with different profiles. Based on these results, the presence of class 1 and 2 integrons was now investigated in these UPEC isolates in order to analyze the distribution of integrons among the phylogenetic groups and virulence profiles. As detected by PCR reactions targeted to the corresponding integrase genes, the class 1 integrons prevailed (22%) followed by those of class 2 (8%). Integrons distributed unevenly among the four main E. coli phylogenetic groups: class 1 integrons predominated in the isolates belonging to group D while class 2 were almost absent in this group. In relation to virulence, integrons frequently appeared in some virulence profiles and were particularly scarce in others. Concerning the class 1 integrons, the most notable findings were that they highly concentrated in isolates presenting one of the virulence profiles (profile V) and were absent in isolates bearing the K1 capsule. The analysis of the Pc promoter variants of the class 1 integrons revealed that all isolates with virulence profile V contained the same Pc version; PcH1. Findings in this work support the idea that, among UPEC strains, integrons would encounter constraints for their installation in some genetic backgrounds while other backgrounds would be propitious for their permanence. PMID:25448130

Poey, María Eloisa; Laviña, Magela

2014-12-01

46

Pilicide ec240 Disrupts Virulence Circuits in Uropathogenic Escherichia coli  

PubMed Central

ABSTRACT Chaperone-usher pathway (CUP) pili are extracellular organelles produced by Gram-negative bacteria that mediate bacterial pathogenesis. Small-molecule inhibitors of CUP pili, termed pilicides, were rationally designed and shown to inhibit type 1 or P piliation. Here, we show that pilicide ec240 decreased the levels of type 1, P, and S piliation. Transcriptomic and proteomic analyses using the cystitis isolate UTI89 revealed that ec240 dysregulated CUP pili and decreased motility. Paradoxically, the transcript levels of P and S pilus genes were increased during growth in ec240, even though the level of P and S piliation decreased. In contrast, the most downregulated transcripts after growth in ec240 were from the type 1 pilus genes. Type 1 pilus expression is controlled by inversion of the fimS promoter element, which can oscillate between phase on and phase off orientations. ec240 induced the fimS phase off orientation, and this effect was necessary for the majority of ec240’s inhibition of type 1 piliation. ec240 increased levels of the transcriptional regulators SfaB and PapB, which were shown to induce the fimS promoter phase off orientation. Furthermore, the effect of ec240 on motility was abolished in the absence of the SfaB, PapB, SfaX, and PapX regulators. In contrast to the effects of ec240, deletion of the type 1 pilus operon led to increased S and P piliation and motility. Thus, ec240 dysregulated several uropathogenic Escherichia coli (UPEC) virulence factors through different mechanisms and independent of its effects on type 1 pilus biogenesis and may have potential as an antivirulence compound. PMID:25352623

Greene, Sarah E.; Pinkner, Jerome S.; Chorell, Erik; Dodson, Karen W.; Shaffer, Carrie L.; Conover, Matt S.; Livny, Jonathan; Hadjifrangiskou, Maria; Almqvist, Fredrik

2014-01-01

47

Molecular regulation of urothelial renewal and host defenses during infection with uropathogenic Escherichia coli.  

PubMed

Uropathogenic Escherichia coli (UPEC), the principal cause of urinary tract infection in women, attaches to the superficial facet cell layer of the bladder epithelium (urothelium) via its FimH adhesin. Attachment triggers exfoliation of bacteria-laden superficial facet cells, followed by rapid reconstitution of the urothelium through differentiation of underlying basal and intermediate cells. We have used DNA microarrays to define the molecular regulators of urothelial renewal and host defense expressed in adult C57Bl/6 female mice during the early phases of infection with isogenic virulent (FimH+) or avirulent (FimH-) UPEC strains. The temporal evolution and cellular origins of selected responses were then characterized by real time quantitative reverse transcriptase-PCR, in situ hybridization, and immunohistochemical analyses. Well before exfoliation is evident, FimH-mediated attachment suppresses transforming growth factor-beta (Bmp4) and Wnt5a/Ca(2+) signaling to promote subsequent differentiation of basal/intermediate cells. The early transcriptional responses to attachment also include induction of regulators of proliferation (e.g. epidermal growth factor family members), induction of the ETS transcription factor Elf3, which transactivates genes involved in epithelial differentiation and host defense (inducible nitric-oxide synthase), induction of modulators, and mediators of pro-inflammatory responses (e.g. Socs3, Cebp/delta, Bcl3, and CC/CXC chemokines), induction of modulators of apoptotic responses (A20), and induction of intermediate cell tight junction components (claudin-4). Both early and late phases of the host response exhibit remarkable specificity for the FimH+ strain and provide new insights about the molecular cascade mobilized to combat UPEC-associated urinary tract infection. PMID:11744708

Mysorekar, Indira U; Mulvey, Matthew A; Hultgren, Scott J; Gordon, Jeffrey I

2002-03-01

48

Suppression of type 1 pilus assembly in uropathogenic Escherichia coli by chemical inhibition of subunit polymerization  

PubMed Central

Objectives To identify and to characterize small-molecule inhibitors that target the subunit polymerization of the type 1 pilus assembly in uropathogenic Escherichia coli (UPEC). Methods Using an SDS–PAGE-based assay, in silico pre-filtered small-molecule compounds were screened for specific inhibitory activity against the critical subunit polymerization step of the chaperone–usher pathway during pilus biogenesis. The biological activity of one of the compounds was validated in assays monitoring UPEC type 1 pilus biogenesis, type 1 pilus-dependent biofilm formation and adherence to human bladder epithelial cells. The time dependence of the in vivo inhibitory activity and the overall effect of the compound on UPEC growth were determined. Results N-(4-chloro-phenyl)-2-{5-[4-(pyrrolidine-1-sulfonyl)-phenyl]-[1,3,4]oxadiazol-2-yl sulfanyl}-acetamide (AL1) inhibited in vitro pilus subunit polymerization. In bacterial cultures, AL1 disrupted UPEC type 1 pilus biogenesis and pilus-dependent biofilm formation, and resulted in the reduction of bacterial adherence to human bladder epithelial cells, without affecting bacterial cell growth. Bacterial exposure to the inhibitor led to an almost instantaneous loss of type 1 pili. Conclusions We have identified and characterized a small molecule that interferes with the assembly of type 1 pili. The molecule targets the polymerization step during the subunit incorporation cycle of the chaperone–usher pathway. Our discovery provides new insight into the design and development of novel anti-virulence therapies targeting key virulence factors of bacterial pathogens. PMID:24324225

Lo, Alvin W. H.; Van de Water, Karen; Gane, Paul J.; Chan, A.W. Edith; Steadman, David; Stevens, Kiri; Selwood, David L.; Waksman, Gabriel; Remaut, Han

2014-01-01

49

Genome-Wide Detection of Fitness Genes in Uropathogenic Escherichia coli during Systemic Infection  

PubMed Central

Uropathogenic Escherichia coli (UPEC) is a leading etiological agent of bacteremia in humans. Virulence mechanisms of UPEC in the context of urinary tract infections have been subjected to extensive research. However, understanding of the fitness mechanisms used by UPEC during bacteremia and systemic infection is limited. A forward genetic screen was utilized to detect transposon insertion mutants with fitness defects during colonization of mouse spleens. An inoculum comprised of 360,000 transposon mutants in the UPEC strain CFT073, cultured from the blood of a patient with pyelonephritis, was used to inoculate mice intravenously. Transposon insertion sites in the inoculum (input) and bacteria colonizing the spleen (output) were identified using high-throughput sequencing of transposon-chromosome junctions. Using frequencies of representation of each insertion mutant in the input and output samples, 242 candidate fitness genes were identified. Co-infection experiments with each of 11 defined mutants and the wild-type strain demonstrated that 82% (9 of 11) of the tested candidate fitness genes were required for optimal fitness in a mouse model of systemic infection. Genes involved in biosynthesis of poly-N-acetyl glucosamine (pgaABCD), major and minor pilin of a type IV pilus (c2394 and c2395), oligopeptide uptake periplasmic-binding protein (oppA), sensitive to antimicrobial peptides (sapABCDF), putative outer membrane receptor (yddB), zinc metallopeptidase (pqqL), a shikimate pathway gene (c1220) and autotransporter serine proteases (pic and vat) were further characterized. Here, we report the first genome-wide identification of genes that contribute to fitness in UPEC during systemic infection in a mammalian host. These fitness factors may represent targets for developing novel therapeutics against UPEC. PMID:24339777

Subashchandrabose, Sargurunathan; Smith, Sara N.; Spurbeck, Rachel R.; Kole, Monica M.; Mobley, Harry L. T.

2013-01-01

50

PafR, a novel transcription regulator, is important for pathogenesis in uropathogenic Escherichia coli.  

PubMed

The metV genomic island in the chromosome of uropathogenic Escherichia coli (UPEC) encodes a putative transcription factor and a sugar permease of the phosphotransferase system (PTS), which are predicted to compose a Bgl-like sensory system. The presence of these two genes, hereby termed pafR and pafP, respectively, has been previously shown to correlate with isolates causing clinical syndromes. We show here that deletion of both genes impairs the ability of the resulting mutant to infect the CBA/J mouse model of ascending urinary tract infection compared to that of the parent strain, CFT073. Expressing the two genes in trans in the two-gene knockout mutant complemented full virulence. Deletion of either gene individually generated the same phenotype as the double knockout, indicating that both pafR and pafP are important to pathogenesis. We screened numerous environmental conditions but failed to detect expression from the promoter that precedes the paf genes in vitro, suggesting that they are in vivo induced (ivi). Although PafR is shown here to be capable of functioning as a transcriptional antiterminator, its targets in the UPEC genome are not known. Using microarray analysis, we have shown that expression of PafR from a heterologous promoter in CFT073 affects expression of genes related to bacterial virulence, biofilm formation, and metabolism. Expression of PafR also inhibits biofilm formation and motility. Taken together, our results suggest that the paf genes are implicated in pathogenesis and that PafR controls virulence genes, in particular biofilm formation genes. PMID:25069986

Baum, Mordechai; Watad, Mobarak; Smith, Sara N; Alteri, Christopher J; Gordon, Noa; Rosenshine, Ilan; Mobley, Harry L; Amster-Choder, Orna

2014-10-01

51

Virulence Plasmid Harbored by Uropathogenic Escherichia coli Functions in Acute Stages of Pathogenesis?  

PubMed Central

Urinary tract infections (UTIs), the majority of which are caused by uropathogenic Escherichia coli (UPEC), afflict nearly 60% of women within their lifetimes. Studies in mice and humans have revealed that UPEC strains undergo a complex pathogenesis cycle that involves both the formation of intracellular bacterial communities (IBC) and the colonization of extracellular niches. Despite the commonality of the UPEC pathogenesis cycle, no specific urovirulence genetic profile has been determined; this is likely due to the fluid nature of the UPEC genome as the result of horizontal gene transfer and numerous genes of unknown function. UTI89 has a large extrachromosomal element termed pUTI89 with many characteristics of UPEC pathogenicity islands and that likely arose due to horizontal gene transfer. The pUTI89 plasmid has characteristics of both F plasmids and other known virulence plasmids. We sought to determine whether pUTI89 is important for virulence. Both in vitro and in vivo assays were used to examine the function of pUTI89 using plasmid-cured UTI89. No differences were observed between UTI89 and plasmid-cured UTI89 based on growth, type 1 pilus expression, or biofilm formation. However, in a mouse model of UTI, a significant decrease in bacterial invasion, CFU and IBC formation of the pUTI89-cured strain was observed at early time points postinfection compared to the wild type. Through directed deletions of specific operons on pUTI89, the cjr operon was partially implicated in this observed defect. Our findings implicate pUTI89 in the early aspects of infection. PMID:20123719

Cusumano, Corinne K.; Hung, Chia S.; Chen, Swaine L.; Hultgren, Scott J.

2010-01-01

52

HlyB-dependent secretion of hemolysin by uropathogenic Escherichia coli requires conserved sequences flanking the chromosomal hly determinant.  

PubMed Central

The synthesis and secretion of hemolysin (HlyA) by Escherichia coli are governed by four contiguous genes (hlyCABD) that are closely conserved on plasmids and, among human pathogenic strains, on the chromosome. We have previously shown that in plasmid pHly152 the coexpressed synthesis and export functions are uncoupled by intraoperon transcription termination, which is in turn alleviated by antitermination dictated in cis by a region upstream of the hly operon. In this study we describe an analogous region of ca. 1,100 base pairs flanking the chromosomal hly determinant of the uropathogenic strain E. coli 2001. This region had no significant effect on intracellular levels of hemolysin but activated strongly, both in cis and in trans, the specific hlyB-hlyD-dependent hemolysin secretion function. The secretion-activating region increased the transcription of the secretion gene hlyB, but the transcription effect was not as pronounced as that seen in the pHly152 determinant and was not evident when the region was present in trans to the hemolysin genes, suggesting that, in addition to transcriptional activation, the region may possibly exert a secondary posttranscriptional influence. Southern hybridizations with the 1,100-base pairs secretion-activating sequence showed low identity to plasmid pHly152 and no identity with total DNA from nonhemolytic uropathogenic E. coli or hemolytic isolates of Proteus vulgaris, Proteus mirabilis, and Morganella morganii. In contrast, hybridization to total DNA from hemolytic E. coli isolates belonging to different serotypes showed strong conservation of the activating sequence, indicating that it is an integral component of the chromosomal hly determinant that is widespread among uropathogenic E. coli. Images FIG. 2 FIG. 3 FIG. 4 FIG. 5 FIG. 6 FIG. 7 PMID:1689714

Cross, M A; Koronakis, V; Stanley, P L; Hughes, C

1990-01-01

53

Molecular Characterization of Uropathogenic Escherichia coli: Nalidixic Acid and Ciprofloxacin Resistance, Virulent Factors and Phylogenetic Background  

PubMed Central

Background and Objective: A proficient pathogen should be virulent, resistant to antibiotics, and epidemic. However, the interplay between resistance and virulence is poorly understood. Perhaps, the most commonly accepted view is that resistance to quinolones is linked to a loss of virulence factors. However, the low virulent phylogenetic groups may be more prone to acquire resistance to quinolones. The aim of this study was to identify and characterise the Nalidixic Acid (NA) and ciprofloxacin (CIP) resistant uropathogenic Escherichia coli (UPEC) isolates with respect to virulence and phylogenetic background, from hospital settings in Kolkata, an eastern region in India. Research based on these bacterial populations will help in understanding the molecular mechanisms underlying the association between resistance and virulence, that in turn, may help in managing the future disseminations of UTIs in their entirety. Material and Methods: One hundred and ten E. coli isolates were screened against NA and CIP using Kirby-Bauer disk diffusion technique, following CLSI guidelines. Prevalence of virulent factor genes and distribution of phylogenetic groups amongst the isolates was determined by PCR, using gene specific primers against the different virulent factors and DNA markers (chuA, yjaA and DNA fragment, TSPE4.C2) respectively. Statistical analysis of the data was performed using SPSS software. Results: Resistance to both NA and CIP was reported in 75.5 % of the isolates which were analysed. The virulent determinants, papC, pap GII, papEF, afa, cnf1, hlyA and iroN were significantly predominant in the drug susceptible than the resistant isolates. A significant reduction of phylogroup B2 in NA (85.7% versus 64.6%, ?2P<0.001) and CIP (85.2 % versus 61.4%, ?2P<0.001) resistant UPEC isolates, followed by increase in predominance of non-B2 phylotypes (group D and group B1), were observed. Conclusion: This is the first report from India that has indicated possible evidence on horizontal gene transfer from pathogenic to commensal strains and selection of the latter, on extensive usage of this group of antimicrobials in hospital settings, where these drugs were routinely prescribed for treating urinary tract infection. Therefore, this information necessitates surveillance programs and administration of effective strategies, to put an end to random prescription policies involving this group of antimicrobials. PMID:24551624

Basu, Shreya; Mukherjee, Sandip Kumar; Hazra, Avijit; Mukherjee, Mandira

2013-01-01

54

Cloning of fimH and fliC and expression of the fusion protein FimH/FliC from Uropathogenic Escherichia coli (UPEC) isolated in Iran  

E-print Network

is isolated in around 80% of UTI cases. Antibiotic therapy resulted in the emergence of antibiotic resistance. ABSTRACT Background and Objectives: Urinary tract infection (UTI) is one of the most common infections in the world. The majority of UTIs are caused by Uropathogenic Escherichia coli (UPEC) strains. FimH and Fli

Paris-Sud XI, Université de

55

Identification of genes subject to positive selection in uropathogenic strains of Escherichia coli: A comparative genomics approach  

PubMed Central

Escherichia coli is a model laboratory bacterium, a species that is widely distributed in the environment, as well as a mutualist and pathogen in its human hosts. As such, E. coli represents an attractive organism to study how environment impacts microbial genome structure and function. Uropathogenic E. coli (UPEC) must adapt to life in several microbial communities in the human body, and has a complex life cycle in the bladder when it causes acute or recurrent urinary tract infection (UTI). Several studies designed to identify virulence factors have focused on genes that are uniquely represented in UPEC strains, whereas the role of genes that are common to all E. coli has received much less attention. Here we describe the complete 5,065,741-bp genome sequence of a UPEC strain recovered from a patient with an acute bladder infection and compare it with six other finished E. coli genome sequences. We searched 3,470 ortholog sets for genes that are under positive selection only in UPEC strains. Our maximum likelihood-based analysis yielded 29 genes involved in various aspects of cell surface structure, DNA metabolism, nutrient acquisition, and UTI. These results were validated by resequencing a subset of the 29 genes in a panel of 50 urinary, periurethral, and rectal E. coli isolates from patients with UTI. These studies outline a computational approach that may be broadly applicable for studying strain-specific adaptation and pathogenesis in other bacteria. PMID:16585510

Chen, Swaine L.; Hung, Chia-Seui; Xu, Jian; Reigstad, Christopher S.; Magrini, Vincent; Sabo, Aniko; Blasiar, Darin; Bieri, Tamberlyn; Meyer, Rekha R.; Ozersky, Philip; Armstrong, Jon R.; Fulton, Robert S.; Latreille, J. Phillip; Spieth, John; Hooton, Thomas M.; Mardis, Elaine R.; Hultgren, Scott J.; Gordon, Jeffrey I.

2006-01-01

56

Occurrence of Antibiotic-Resistant Uropathogenic Escherichia coli Clonal Group A in Wastewater Effluents  

Microsoft Academic Search

Received 21 September 2006\\/Accepted 27 April 2007 Isolates of Escherichia coli belonging to clonal group A (CGA), a recently described disseminated cause of drug-resistant urinary tract infections in humans, were present in four of seven sewage effluents collected from geographically dispersed areas of the United States. All 15 CGA isolates (1% of the 1,484 isolates analyzed) exhibited resistance to trimethoprim-sulfamethoxazole

Laura A. Boczek; Eugene W. Rice; Brian Johnston; James R. Johnson

2007-01-01

57

The Association of Virulence Determinants of Uropathogenic Escherichia coli With Antibiotic Resistance  

PubMed Central

Background: The emergence of antimicrobial resistant strains of Escherichia coli has raised considerable interest in understanding the diversity and epidemiology of E. coli infections in humans. Virulence factors of E. coli determine the specific infections caused by this microorganism. Objectives: This study aimed to determine the prevalence of eight E. coli virulence factors and their association with antimicrobial resistance in bacteria isolated from patients with urinary tract infections (UTI). Patients and Methods: One thousand patients with UTI were enrolled in this cross-sectional study. Antimicrobial susceptibility was examined by disc diffusion method according to CLSI guidelines. After DNA extraction, the materials were probed by PCR for eight virulence factors genes, namely fimH, hly, iucC, ibeA, sfa/foc, neuC, papC, and afa genes. Results: The frequency of virulence factors papC, afa, sfa/foc, fimH, hly, neuC, ibeA, and iucC were 53.3%, 51.7%, 53.3%, 56.7%, 23.3%, 31.7%, 20%, and 73.3%, respectively. In addition, there was a high degree resistance to cotrimoxazole and nalidixic acid while a high degree of susceptibility to nitrofurantoin was detected. There was a statistically significant association between fimH gene and resistance to ciprofloxacin (P = 0.006), nalidixic acid (P = 0.025), and cotrimoxazole (P = 0.02). Such associations were found between ibeA gene and amikacin (P = 0.02) and cotrimoxazole (P = 0.02) as well as between afa gene and gentamycin (P = 0.05). Conclusions: The results showed that E. coli isolated from patients with UTI had eight virulence factors with high frequencies. Moreover, these results alleged a direct connection between virulence factors and antimicrobial resistance in E. coli. PMID:25147722

Asadi, Sara; Kargar, Mohammad; Solhjoo, Kavous; Najafi, Akram; Ghorbani-Dalini, Sadegh

2014-01-01

58

Antibiotic resistance profile and virulence genes of uropathogenic Escherichia coli isolates in relation to phylogeny.  

PubMed

Escherichia coli (E. coli) strains are the major cause of urinary tract infections (UTI) and belong to the large group of extra-intestinal pathogenic E. coli. The purposes of this study were to determine the antibiotic resistance profile, virulence genes and phylogenetic background of E. coli isolates from UTI cases. A total of 137 E. coli isolates were obtained from UTI samples. The antimicrobial susceptibility of confirmed isolates was determined by disk diffusion method against eight antibiotics. The isolates were examined to determine the presence and prevalence of selected virulence genes including iucD, sfa/focDE, papEF and hly. ECOR phylo-groups of isolates were determined by detection of yjaA and chuA genes and fragment TspE4.C2. The antibiogram results showed that 71% of the isolates were resistant to cefazolin, 60.42% to co-trimoxazole, 54.16% to nalidixic acid, 36.45% to gentamicin, 29.18% to ciprofloxacin, 14.58% to cefepime, 6.25% to nitrofurantoin and 0.00% to imipenem. Twenty-two antibiotic resistance patterns were observed among the isolates. Virulence genotyping of isolates revealed that 58.39% isolates had at least one of the four virulence genes. The iucD gene was the most prevalent gene (43.06%). The other genes including sfa/focDE, papEF and hly genes were detected in 35.76%, 18.97% and 2.18% isolates, respectively. Nine combination patterns of the virulence genes were detected in isolates. Phylotyping of 137 isolates revealed that the isolates fell into A (45.99%), B1 (13.14%), B2 (19.71%) and D (21.16%) groups. Phylotyping of multidrug resistant isolates indicated that these isolates are mostly in A (60.34%) and D (20.38%) groups. In conclusion, the isolates that possessed the iucD, sfa/focDE, papEF and hly virulence genes mostly belonged to A and B2 groups, whereas antibiotic resistant isolates were in groups A and D. Escherichia coli strains carrying virulence factors and antibiotic resistance are distributed in specific phylogenetic background. PMID:24862040

Adib, N; Ghanbarpour, R; Solatzadeh, H; Alizade, H

2014-03-01

59

Virulence Characteristics and Genetic Affinities of Multiple Drug Resistant Uropathogenic Escherichia coli from a Semi Urban Locality in India  

PubMed Central

Extraintestinal pathogenic Escherichia coli (ExPEC) are of significant health concern. The emergence of drug resistant E. coli with high virulence potential is alarming. Lack of sufficient data on transmission dynamics, virulence spectrum and antimicrobial resistance of certain pathogens such as the uropathogenic E. coli (UPEC) from countries with high infection burden, such as India, hinders the infection control and management efforts. In this study, we extensively genotyped and phenotyped a collection of 150 UPEC obtained from patients belonging to a semi-urban, industrialized setting near Pune, India. The isolates representing different clinical categories were analyzed in comparison with 50 commensal E. coli isolates from India as well as 50 ExPEC strains from Germany. Virulent strains were identified based on hemolysis, haemagglutination, cell surface hydrophobicity, serum bactericidal activity as well as with the help of O serotyping. We generated antimicrobial resistance profiles for all the clinical isolates and carried out phylogenetic analysis based on repetitive extragenic palindromic (rep)-PCR. E. coli from urinary tract infection cases expressed higher percentages of type I (45%) and P fimbriae (40%) when compared to fecal isolates (25% and 8% respectively). Hemolytic group comprised of 60% of UPEC and only 2% of E. coli from feces. Additionally, we found that serum resistance and cell surface hydrophobicity were not significantly (p?=?0.16/p?=?0.51) associated with UPEC from clinical cases. Moreover, clinical isolates exhibited highest resistance against amoxicillin (67.3%) and least against nitrofurantoin (57.3%). We also observed that 31.3% of UPEC were extended-spectrum beta-lactamase (ESBL) producers belonging to serotype O25, of which four were also positive for O25b subgroup that is linked to B2-O25b-ST131-CTX-M-15 virulent/multiresistant type. Furthermore, isolates from India and Germany (as well as global sources) were found to be genetically distinct with no evidence to espouse expansion of E. coli from India to the west or vice-versa. PMID:21464963

Kumar, Ashutosh; Parveen, Sana; Gandham, Nageshwari; Wieler, Lothar H.; Ewers, Christa; Ahmed, Niyaz

2011-01-01

60

The Co-Transcriptome of Uropathogenic Escherichia coli-Infected Mouse Macrophages Reveals New Insights into Host-Pathogen Interactions.  

PubMed

Urinary tract infections (UTI) are among the most common infections in humans. Uropathogenic Escherichia coli (UPEC) can invade and replicate within bladder epithelial cells, and some UPEC strains can also survive within macrophages. To understand the UPEC transcriptional program associated with intramacrophage survival, we performed host-pathogen co-transcriptome analyses using RNA sequencing. Mouse bone marrow-derived macrophages (BMMs) were challenged over a 24 h time course with two UPEC reference strains that possess contrasting intramacrophage phenotypes: UTI89, which survives in BMMs, and 83972, which is killed by BMMs. Neither of these strains caused significant BMM cell death at the low multiplicity of infection that was used in this study. We developed an effective computational framework that simultaneously separated, annotated, and quantified the mammalian and bacterial transcriptomes. BMMs responded to the two UPEC strains with a broadly similar gene expression program. In contrast, the transcriptional responses of the UPEC strains diverged markedly from each other. We identified UTI89 genes upregulated at 24 h post-infection, and hypothesized that some may contribute to intramacrophage survival. Indeed, we showed that deletion of one such gene (pspA) significantly reduced UTI89 survival within BMMs. Our study provides a technological framework for simultaneously capturing global changes at the transcriptional level in co-cultures, and has generated new insights into the mechanisms that UPEC use to persist within the intramacrophage environment. PMID:25410299

Mavromatis, Charalampos Harris; Bokil, Nilesh J; Totsika, Makrina; Kakkanat, Asha; Schaale, Kolja; Cannistraci1, Carlo V; Ryu, Taewoo; Beatson, Scott A; Ulett, Glen C; Schembri, Mark A; Sweet, Matthew J; Ravasi, Timothy

2014-11-20

61

Flagellin/TLR5 signalling activates renal collecting duct cells and facilitates invasion and cellular translocation of uropathogenic Escherichia coli.  

PubMed

Uropathogenic Escherichia coli (UPEC) colonizing kidneys is the main cause of acute pyelonephritis. TLR5 that senses flagellin was shown to be highly expressed in the bladder and to participate in host defence against flagellated UPEC, although its role in kidneys still remains elusive. Here we show that TLR5 is expressed in renal medullary collecting duct (MCD) cells, which represent a preferential site of UPEC adhesion. Flagellin, like lipopolysaccharide, stimulated the production of the chemoattractant chemokines CXCL1 and CXCL2, and subsequent migration capacity of neutrophils in cultured wild-type (WT) and Tlr4(-/-) MCDs, but not in Tlr5(-/-) MCDs. UPEC can translocate across intact MCD layers without altering tight junctions. Strikingly, the invasion capacity and transcellular translocation of the UPEC strain HT7 were significantly lower in Tlr5(-/-) than in WT MCDs. The non-motile HT7?fliC mutant lacking flagellin also exhibited much lower translocation capacities than the HT7 isolates. Finally, Tlr5(-/-) kidneys exhibited less infiltrating neutrophils than WT kidneys one day after the transurethral inoculation of HT7, and greater delayed renal bacterial loads in the day 4 post-infected Tlr5(-/-) kidneys. Overall, these findings indicate that the epithelial TLR5 participates to renal antibacterial defence, but paradoxically favours the translocation of UPEC across intact MCD cell layers. PMID:24779433

Bens, Marcelle; Vimont, Sophie; Ben Mkaddem, Sanae; Chassin, Cécilia; Goujon, Jean-Michel; Balloy, Viviane; Chignard, Michel; Werts, Catherine; Vandewalle, Alain

2014-10-01

62

G-CSF induction early in uropathogenic Escherichia coli infection of the urinary tract modulates host immunity  

PubMed Central

Summary Uropathogenic Escherichia coli (UPEC), the causative agent of approximately 85% of urinary tract infections (UTI), is a major health concern primarily affecting women. During infection, neutrophils infiltrate the bladder, but the mechanism of recruitment is not well understood. Here, we investigated the role of UPEC-induced cytokine production in neutrophil recruitment and UTI progression. We first examined the kinetics of cytokine expression during UPEC infection of the bladder, and their contribution to neutrophil recruitment. We found that UPEC infection induces expression of several pro-inflammatory cytokines including granulocyte colony-stimulating factor (G-CSF, CSF-3), not previously known to be involved in the host response to UTI. G-CSF induces neutrophil emigration from the bone marrow; these cells are thought to be critical for bacterial clearance during infection. Upon neutralization of G-CSF during UPEC infection, we found fewer circulating neutrophils, decreased neutrophil infiltration into the bladder and, paradoxically, a decreased bacterial burden in the bladder. However, depletion of G-CSF resulted in a corresponding increase in macrophage-activating cytokines, such as monocyte chemotactic protein-1 (MCP-1, CCL-2) and Il-1?, which may be key in host response to UPEC infection, potentially resolving the paradoxical decreased bacterial burden. Thus, G-CSF acts in a previously unrecognized role to modulate the host inflammatory response during UPEC infection. PMID:18754853

Ingersoll, Molly A.; Kline, Kimberly A.; Nielsen, Hailyn V.; Hultgren, Scott J.

2010-01-01

63

Virulence genes, quinolone and fluoroquinolone resistance, and phylogenetic background of uropathogenic Escherichia coli strains isolated in Japan.  

PubMed

A total of 312 uropathogenic Escherichia coli strains were isolated from clinical specimens in 7 hospitals in Aichi Prefecture, Japan. Among them, 113 strains were resistant to quinolone, and 49 strains were resistant to fluoroquinolone. Phylogenetic group B2 was most prevalent in both susceptible strains (148 of 199 strains, 74.4%) and resistant strains (quinolone-resistant strains, 73 of 113 strains, 64.6%; fluoroquinolone-resistant strains, 40 of 49 strains, 81.6%). The resistant strains showed a significantly lower prevalence of virulence genes papA, hlyA, and cnf1 than did the susceptible strains, and this observation was further obvious when compared within B2 group strains. Among the 40 fluoroquinolone-resistant strains belonging to group B2, 37 (92.5%) strains carried PAIusp subtype IIa, 36 strains of which carried E84V mutation in parC, whereas none of the 9 strains belonging to group D carried PAIusp subtype IIa, and only one strain carried the mutation. These observations indicate that the differences of phenotypes including resistance of quinolone and carriage of virulence genes are associated with the complex context of genetic background, including the phylogenetic group and PAIusp subtype. PMID:20332573

Kawamura-Sato, Kumiko; Yoshida, Risa; Shibayama, Keigo; Ohta, Michio

2010-03-01

64

Pilicides inhibit the FGL chaperone/usher assisted biogenesis of the Dr fimbrial polyadhesin from uropathogenic Escherichia coli  

PubMed Central

Background The global spread of bacterial resistance has given rise to a growing interest in new anti-bacterial agents with a new strategy of action. Pilicides are derivatives of ring-fused 2-pyridones which block the formation of the pili/fimbriae crucial to bacterial pathogenesis. They impair by means of a chaperone-usher pathway conserved in the Gram-negative bacteria of adhesive structures biogenesis. Pili/fimbriae of this type belong to two subfamilies, FGS and FGL, which differ in the details of their assembly mechanism. The data published to date have shown that pilicides inhibit biogenesis of type 1 and P pili of the FGS type which are encoded by uropathogenic E. coli strains. Results We evaluated the anti-bacterial activity of literature pilicides as blockers of the assembly of a model example of FGL-type adhesive structures, – the Dr fimbriae encoded by a dra gene cluster of uropathogenic Escherichia coli strains. In comparison to the strain grown without pilicide, the Dr+ bacteria cultivated in the presence of the 3.5 mM concentration of pilicides resulted in a reduction of 75 to 87% in the adherence properties to CHO cells expressing Dr fimbrial DAF receptor protein. Using quantitative assays, we determined the amount of Dr fimbriae in the bacteria cultivated in the presence of 3.5 mM of pilicides to be reduced by 75 to 81%. The inhibition effect of pilicides is concentration dependent, which is a crucial property for their use as potential anti-bacterial agents. The data presented in this article indicate that pilicides in mM concentration effectively inhibit the adherence of Dr+ bacteria to the host cells, – the crucial, initial step in bacterial pathogenesis. Conclusions Structural analysis of the DraB chaperone clearly showed it to be a model of the FGL subfamily of chaperones. This permits us to conclude that analyzed pilicides in mM concentration are effective inhibitors of the assembly of adhesins belonging to the Dr family, and more speculatively, of other FGL-type adhesive organelles. The presented data and those published so far permit to speculate that based on the conservation of chaperone-usher pathway in Gram-negative bacteria , the pilicides are potential anti-bacterial agents with activity against numerous pathogens, the virulence of which is dependent on the adhesive structures of the chaperone-usher type. PMID:23758700

2013-01-01

65

Haemolytic Escherichia coli isolated from dogs with diarrhea have characteristics of both uropathogenic and necrotoxigenic strains.  

PubMed

Twenty-four haemolytic Escherichia coli strains were isolated from dogs with diarrhea. The strains were serotyped and analysed by polymerase chain reaction (PCR) for genes encoding virulence factors associated with E. coli that cause diarrhea in animals. Adhesion antigen production was deduced from haemagglutination experiments. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) of heat extracts was also used as an indication for the production of adhesive structures. The majority of the strains was shown to produce this type of virulence factor. Adhesion and invasion tests of the strains and Caco-2 cells showed that all strains adhered and that two were invasive. The two invasive strains were positive in the intimin PCR and one of them also contained genes encoding CS31A. The PCR for heat stable toxin (ST) was positive in only four strains, as was the presence of F17 fimbrial genes. Surprisingly, 19 strains had intact P fimbrial operons, coding for an adhesin involved in urinary tract infection (UTI). The cytotoxic necrotising factor 1 (CNF1) gene, also mainly found in UTI was likewise detected in these 19 strains. Cytolethal distending toxin (Cdt) genes were found in five strains. The high number of strains positive for CNF1 and P fimbriae prompted us to test the strains in a multiplex PCR used to test E. coli isolated from UTI in various species for 30 virulence associated genes. The data showed that the majority of the diarrhea isolates have virulence factor profiles highly similar to UTI E. coli isolates from dogs. This raises the question whether these isolates are real intestinal pathogens or "innocent bystanders". However, since CNF1 producing necrotoxic E. coli (NTEC) strains isolated from humans, pigs and calves with diarrhea appear to be highly related to our strains, it might be that in dogs this type of isolate is capable of causing not only UTI, but also diarrhea. If this is the case and this type of isolate is "bifunctional", domestic animals likely constitute a reservoir of NTEC strains which can be also pathogenic for humans. PMID:11856586

Starcic, Marjanca; Johnson, James R; Stell, Adam L; van der Goot, Jeanet; Hendriks, Henno G C J M; van Vorstenbosch, Camillo; van Dijk, Linda; Gaastra, Wim

2002-04-01

66

Quantification of filamentation by uropathogenic Escherichia coli during experimental bladder cell infection by using semi-automated image analysis.  

PubMed

Several rod-shaped pathogens including Escherichia coli, Salmonella spp. and Klebsiella pneumonia are capable of adopting highly filamentous cell shapes under certain circumstances. This phenomenon occurs as a result of continued cell elongation during growth without the usual septation into single rod-shaped cells. Evidence has emerged over the past decade suggesting that this morphological transformation is controlled and reversible and provides selective advantages under certain growth conditions, such as during infection in humans. In order to identify the factors which induce filamentation of bacterial pathogens and study the advantages of bacterial morphological plasticity, methods are needed to accurately quantify changes in bacterial cell shape. In this study, we present a method for quantification of bacterial filamentation based on automatic detection and measurement of bacterial units in focus-stacked microscopy images. Used in combination with a flow-chamber based in vitro cystitis model, we study the factors involved in filament formation by uropathogenic E. coli (UPEC) during infection. The influence of substratum surface, intracellular proliferation and flow media on UPEC filamentation is evaluated. We show that reversible UPEC filamentation during cystitis is not dependent on intracellular infection, which previous studies have suggested. Instead, we find that filamentation can be induced by contact with surfaces, both biological and artificial. Lastly our data indicate that UPEC filamentation is induced by trace-amounts of specific components in urine, rather than being a generic stress-response to high urine salt concentrations. The study shows that the combined methodology is generally useful for investigation of bacterial morphological transitions during cell infection. PMID:25546841

Klein, Kasper; Palarasah, Yaseelan; Kolmos, Hans Jørn; Møller-Jensen, Jakob; Andersen, Thomas Emil

2015-02-01

67

Toxin-Antitoxin Systems Are Important for Niche-Specific Colonization and Stress Resistance of Uropathogenic Escherichia coli  

PubMed Central

Toxin-antitoxin (TA) systems are prevalent in many bacterial genomes and have been implicated in biofilm and persister cell formation, but the contribution of individual chromosomally encoded TA systems during bacterial pathogenesis is not well understood. Of the known TA systems encoded by Escherichia coli, only a subset is associated with strains of extraintestinal pathogenic E. coli (ExPEC). These pathogens colonize diverse niches and are a major cause of sepsis, meningitis, and urinary tract infections. Using a murine infection model, we show that two TA systems (YefM-YoeB and YbaJ-Hha) independently promote colonization of the bladder by the reference uropathogenic ExPEC isolate CFT073, while a third TA system comprised of the toxin PasT and the antitoxin PasI is critical to ExPEC survival within the kidneys. The PasTI TA system also enhances ExPEC persister cell formation in the presence of antibiotics and markedly increases pathogen resistance to nutrient limitation as well as oxidative and nitrosative stresses. On its own, low-level expression of PasT protects ExPEC from these stresses, whereas overexpression of PasT is toxic and causes bacterial stasis. PasT-induced stasis can be rescued by overexpression of PasI, indicating that PasTI is a bona fide TA system. By mutagenesis, we find that the stress resistance and toxic effects of PasT can be uncoupled and mapped to distinct domains. Toxicity was specifically linked to sequences within the N-terminus of PasT, a region that also promotes the development of persister cells. These results indicate discrete, multipurpose functions for a TA-associated toxin and demonstrate that individual TA systems can provide bacteria with pronounced fitness advantages dependent on toxin expression levels and the specific environmental niche occupied. PMID:23055930

Norton, J. Paul; Mulvey, Matthew A.

2012-01-01

68

[Prevalence and characterization of uropathogenic Escherichia coli producing cytotoxic necrotizing factor type 1 (CNF1) in the center of Tunisia].  

PubMed

During a three-year survey of the prevalence in central Tunisia of Escherichia coli producing CNF1 toxin (NTEC), 716 samples have been investigated by PCR for cnf1 gene. All samples were isolated from urine of adult and children patients presenting significant bacteriuria (> 10(5) colony-forming units/mL), independently of the severity of the clinical presentation; 328 strains were found harboring cnf1 gene, they were distributed into three clinical categories: 219 (66.76%) from patients with symptomatic bacteriuria, 76 (23.17%) from patients with uncomplicated cystitis and 33 (10.06%) from patients with acute pyelonephritis and complicated urinary tract infections. 98.78% (324) of CNF1 strains presented hemolytic activity. All 328 CNF1 strains harbored both sfa and pap genes and expressed MRHA activity. They belonged to 16 different serotypes. The most common serotypes, in order of frequency, were O6 (25.91%), O4 (17.98%), O2 (12.5%), O75 (9.14%), O78 (8.35%), and O83 (3.65%). Two strains (0.6%) were O168; a serotype shown to be associated to CNF2 producing bovine strains. The frequency of uropathogenic CNF1 strains in center Tunisia was about 45.81% and increased from 26.08% in 1998 to 58.16% in 2000. We showed that E. coli producing cytotoxic necrotizing factor (CNF1) was implicated in urinary tract infections in center Tunisia but no difference was shown between strains isolated from patients with complicated or uncomplicated urinary tract infections. The presence of CNF1 toxin with various associated virulence factors seemed to increase the risk for severe forms of urinary tract infections. PMID:15217764

Denden, I; Mahdouani, K; Bakhrouf, A

2004-01-01

69

Virulence genes and P fimbriae PapA subunit diversity in canine and feline uropathogenic Escherichia coli.  

PubMed

In this study, a total of 118 Escherichia coli strains isolated from dogs (93) and cats (25) with urinary tract infection (UTI) were tested in a multiplex polymerase chain reaction for the presence of adhesin-encoding genes (pap, sfa, and afa), hemolysin encoding genes (hly), cytotoxic necrotizing factor 1 (cnf1) and aerobactin (aer) genes. Virulence gene frequencies detected in those isolates which had been randomly collected (68 canine strains) were: 43% pap, 57% sfa, 1% afa, 44% hly, 41% cnf1 and 34% aer. These frequencies were much higher in the remaining 50 hemolytic strains of either cat or dog origin. Virulence factor associations in the 80 hemolytic strains studied revealed that 50/80 simultaneously had two adhesin genes (pap and sfa) and two cytotoxin genes (hly and cnf1), and 15/80 in addition had the aer gene. The major structural subunit and antigenic determinant of P fimbriae of uropathogenic E. coli is PapA. Polymorphism in this subunit was studied by an F antigen-specific papA allele polymerase chain reaction in 51 canine and 22 feline pap positive E. coli strains. The most prevalent canine papA alleles were F10 (39%), F15 (37%) and F12 (35%). In feline strains F15 (50%) was more frequent, other allele frequencies were F12 (45%), F14 and F10 (27%) and F16 (23%). Only nine canine and two feline strains were negative for one of the 11 serologically defined F types of P fimbriae. Three copies of the pap operon were found in 16/51 canine and 9/22 feline UTI E. coli pap positive strains. In this study, we show that a particular combination of virulence genes appears with high frequency in dog and cat urinary tract E. coli strains (pap, sfa, hly, and cnf1). In spite of the more frequent presence of F10, F12 and F15 papA alleles in this virulence gene combination, the occurrence of different papA alleles in strains where up to three copies of the pap operon are present accounts for the observed P fimbriae diversity. PMID:11423198

Féria, C; Machado, J; Correia, J D; Gonçalves, J; Gaastra, W

2001-09-01

70

Crystal structure of c5321: a protective antigen present in uropathogenic Escherichia coli strains displaying an SLR fold  

PubMed Central

Background Increasing rates of antimicrobial resistance among uropathogens led, among other efforts, to the application of subtractive reverse vaccinology for the identification of antigens present in extraintestinal pathogenic E. coli (ExPEC) strains but absent or variable in non-pathogenic strains, in a quest for a broadly protective Escherichia coli vaccine. The protein coded by locus c5321 from CFT073 E. coli was identified as one of nine potential vaccine candidates against ExPEC and was able to confer protection with an efficacy of 33% in a mouse model of sepsis. c5321 (known also as EsiB) lacks functional annotation and structurally belongs to the Sel1-like repeat (SLR) family. Herein, as part of the general characterization of this potential antigen, we have focused on its structural properties. Results We report the 1.74 Å-resolution crystal structure of c5321 from CFT073 E. coli determined by Se-Met SAD phasing. The structure is composed of 11 SLR units in a topological organisation that highly resembles that found in HcpC from Helicobacter pylori, with the main difference residing in how the super-helical fold is stabilised. The stabilising effect of disulfide bridges in HcpC is replaced in c5321 by a strengthening of the inter-repeat hydrophobic core. A metal-ion binding site, uncharacteristic of SLR proteins, is detected between SLR units 3 and 4 in the region of the inter-repeat hydrophobic core. Crystal contacts are observed between the C-terminal tail of one molecule and the C-terminal amphipathic groove of a neighbouring one, resembling interactions between ligand and proteins containing tetratricopeptide-like repeats. Conclusions The structure of antigen c5321 presents a mode of stabilization of the SLR fold different from that observed in close homologs of known structure. The location of the metal-ion binding site and the observed crystal contacts suggest a potential role in regulation of conformational flexibility and interaction with yet unidentified target proteins, respectively. These findings open new perspectives in both antigen design and for the identification of a functional role for this protective antigen. PMID:24099525

2013-01-01

71

Cytotoxic Necrotizing Factor 1 and Hemolysin from Uropathogenic Escherichia coli Elicit Different Host Responses in the Murine Bladder  

PubMed Central

Cytotoxic necrotizing factor 1 (CNF1) and hemolysin (HlyA1) are toxins produced by uropathogenic Escherichia coli (UPEC). We previously showed that these toxins contribute to the inflammation and tissue damage seen in a mouse model of ascending urinary tract infection. CNF1 constitutively activates small Rho GTPases by deamidation of a conserved glutamine residue, and HlyA1 forms pores in eukaryotic cell membranes. In this study, we used cDNA microarrays of bladder tissue isolated from mice infected intraurethrally with wild-type CP9, CP9cnf1, or CP9?hlyA to further evaluate the role that each toxin plays in the host response to UPEC. Regardless of the strain used, we found that UPEC itself elicited a significant change in host gene expression 24 h after inoculation. The largest numbers of upregulated genes were in the cytokine and chemokine signaling and Toll-like receptor signaling pathways. CNF1 exerted a strong positive influence on expression of genes involved in innate immunity and signal transduction and a negative impact on metabolism- and transport-associated genes. HlyA1 evoked an increase in expression of genes that encode innate immunity factors and a decrease in expression of genes involved in cytoskeletal and metabolic processes. Multiplex cytokine and myeloperoxidase assays corroborated our finding that a strong proinflammatory response was elicited by all strains tested. Bladders challenged intraurethrally with purified CNF1 displayed pathology similar to but significantly less intense than the pathology that we observed in CP9-challenged mice. Our data demonstrate substantial roles for CNF1 and HlyA1 in initiation of a strong proinflammatory response to UPEC in the bladder. PMID:23090961

Garcia, Tamako A.; Ventura, Christy L.; Smith, Mark A.; Merrell, D. Scott

2013-01-01

72

Cytotoxic necrotizing factor 1 and hemolysin from uropathogenic Escherichia coli elicit different host responses in the murine bladder.  

PubMed

Cytotoxic necrotizing factor 1 (CNF1) and hemolysin (HlyA1) are toxins produced by uropathogenic Escherichia coli (UPEC). We previously showed that these toxins contribute to the inflammation and tissue damage seen in a mouse model of ascending urinary tract infection. CNF1 constitutively activates small Rho GTPases by deamidation of a conserved glutamine residue, and HlyA1 forms pores in eukaryotic cell membranes. In this study, we used cDNA microarrays of bladder tissue isolated from mice infected intraurethrally with wild-type CP9, CP9cnf1, or CP9?hlyA to further evaluate the role that each toxin plays in the host response to UPEC. Regardless of the strain used, we found that UPEC itself elicited a significant change in host gene expression 24 h after inoculation. The largest numbers of upregulated genes were in the cytokine and chemokine signaling and Toll-like receptor signaling pathways. CNF1 exerted a strong positive influence on expression of genes involved in innate immunity and signal transduction and a negative impact on metabolism- and transport-associated genes. HlyA1 evoked an increase in expression of genes that encode innate immunity factors and a decrease in expression of genes involved in cytoskeletal and metabolic processes. Multiplex cytokine and myeloperoxidase assays corroborated our finding that a strong proinflammatory response was elicited by all strains tested. Bladders challenged intraurethrally with purified CNF1 displayed pathology similar to but significantly less intense than the pathology that we observed in CP9-challenged mice. Our data demonstrate substantial roles for CNF1 and HlyA1 in initiation of a strong proinflammatory response to UPEC in the bladder. PMID:23090961

Garcia, Tamako A; Ventura, Christy L; Smith, Mark A; Merrell, D Scott; O'Brien, Alison D

2013-01-01

73

Cytotoxic Necrotizing Factor Type 1 of Uropathogenic Escherichia coli Kills Cultured Human Uroepithelial 5637 Cells by an Apoptotic Mechanism  

PubMed Central

Pathogenic Escherichia coli associated with urinary tract infections (UTIs) in otherwise healthy individuals frequently produce cytotoxic necrotizing factor type 1 (CNF1), a member of the family of bacterial toxins that target the Rho family of small GTP-binding proteins. To gain insight into the function of CNF1 in the development of E. coli-mediated UTIs, we examined the effects of CNF1 intoxication on a panel of human cell lines derived from physiologically relevant sites (bladder, ureters, and kidneys). We identified one uroepithelial cell line that exhibited a distinctly different CNF1 intoxication phenotype from the prototypic one of multinucleation without cell death that is seen when HEp-2 or other epithelial cells are treated with CNF1. The 5637 bladder cell line detached from the growth surface within 72 h of CNF1 intoxication, a finding that suggested frank cytotoxicity. To determine the basis for the unexpected toxic effect of CNF1 on 5637 cells, we compared the degree of toxin binding, actin fiber formation, and Rho modification with those CNF1-induced events in HEp-2 cells. We found no apparent difference in the amount of CNF1 bound to 5637 cells and HEp-2 cells. Moreover, CNF1 modified Rho, in vivo and in vitro, in both cell types. In contrast, one of the classic responses to CNF1 in HEp-2 and other epithelial cell lines, the formation of actin stress fibers, was markedly absent in 5637 cells. Indeed, actin stress fiber induction by CNF1 did not occur in any of the other human bladder cell lines that we tested (J82, SV-HUC-1, or T24). Furthermore, the appearance of lamellipodia and filopodia in 5637 cells suggested that CNF1 activated the Cdc42 and Rac proteins. Finally, apoptosis was observed in CNF1-intoxicated 5637 cells. If our results with 5637 cells reflect the interaction of CNF1 with the transitional uroepithelium in the human bladder, then CNF1 may be involved in the exfoliative process that occurs in that organ after infection with uropathogenic E. coli. PMID:10992497

Mills, Melody; Meysick, Karen C.; O'Brien, Alison D.

2000-01-01

74

Uropathogenic Escherichia coli Suppresses the host inflammatory response via pathogenicity island genes sisA and sisB.  

PubMed

Extraintestinal pathogenic Escherichia coli can successfully colonize the urinary tract of the immunocompetent host. In part, this is accomplished by dampening the host immune response. Indeed, the sisA and sisB genes (shiA-like inflammation suppressor genes A and B) of uropathogenic E. coli strain CFT073, homologs of the Shigella flexneri SHI-2 pathogenicity island gene shiA, suppress the host inflammatory response. A double deletion mutant (DeltasisA DeltasisB) resulted in a hyperinflammatory phenotype in an experimental model of ascending urinary tract infection. The DeltasisA DeltasisB mutant not only caused significantly more inflammatory foci in the kidneys of CBA/J mice (P = 0.0399), but these lesions were also histologically more severe (P = 0.0477) than lesions observed in mice infected with wild-type CFT073. This hyperinflammatory phenotype could be suppressed to wild-type levels by in vivo complementation of the DeltasisA DeltasisB mutant with either the sisA or sisB gene in trans. The DeltasisA DeltasisB mutant was outcompeted by wild-type CFT073 during cochallenge infection in the bladder (P = 0.0295) at 48 h postinoculation (hpi). However, during cochallenge infections, we reasoned that wild-type CFT073 could partially complement the DeltasisA DeltasisB mutant. Consistent with this, the most significant colonization defect of the DeltasisA DeltasisB mutant in vivo was observed during independent challenge relative to wild-type CFT073, with attenuation of the mutant observed in the bladder (P < 0.0001) and kidneys (P = 0.0003) at 6 hpi. By 24 and 48 hpi, the DeltasisA DeltasisB mutant was no longer significantly attenuated in the bladder or kidneys, suggesting that the sisA and sisB genes may be important for suppressing the host immune response during the initial stages of infection. PMID:19797063

Lloyd, Amanda L; Smith, Sara N; Eaton, Kathryn A; Mobley, Harry L T

2009-12-01

75

Impact of UV and Peracetic Acid Disinfection on the Prevalence of Virulence and Antimicrobial Resistance Genes in Uropathogenic Escherichia coli in Wastewater Effluents  

PubMed Central

Wastewater discharges may increase the populations of pathogens, including Escherichia coli, and of antimicrobial-resistant strains in receiving waters. This study investigated the impact of UV and peracetic acid (PAA) disinfection on the prevalence of virulence and antimicrobial resistance genes in uropathogenic Escherichia coli (UPEC), the most abundant E. coli pathotype in municipal wastewaters. Laboratory disinfection experiments were conducted on wastewater treated by physicochemical, activated sludge, or biofiltration processes; 1,766 E. coli isolates were obtained for the evaluation. The target disinfection level was 200 CFU/100 ml, resulting in UV and PAA doses of 7 to 30 mJ/cm2 and 0.9 to 2.0 mg/liter, respectively. The proportions of UPECs were reduced in all samples after disinfection, with an average reduction by UV of 55% (range, 22% to 80%) and by PAA of 52% (range, 11% to 100%). Analysis of urovirulence genes revealed that the decline in the UPEC populations was not associated with any particular virulence factor. A positive association was found between the occurrence of urovirulence and antimicrobial resistance genes (ARGs). However, the changes in the prevalence of ARGs in potential UPECs were different following disinfection, i.e., UV appears to have had no effect, while PAA significantly reduced the ARG levels. Thus, this study showed that both UV and PAA disinfections reduced the proportion of UPECs and that PAA disinfection also reduced the proportion of antimicrobial resistance gene-carrying UPEC pathotypes in municipal wastewaters. PMID:24727265

Biswal, Basanta Kumar; Khairallah, Ramzi; Bibi, Kareem; Mazza, Alberto; Gehr, Ronald; Masson, Luke

2014-01-01

76

Impact of UV and peracetic acid disinfection on the prevalence of virulence and antimicrobial resistance genes in uropathogenic Escherichia coli in wastewater effluents.  

PubMed

Wastewater discharges may increase the populations of pathogens, including Escherichia coli, and of antimicrobial-resistant strains in receiving waters. This study investigated the impact of UV and peracetic acid (PAA) disinfection on the prevalence of virulence and antimicrobial resistance genes in uropathogenic Escherichia coli (UPEC), the most abundant E. coli pathotype in municipal wastewaters. Laboratory disinfection experiments were conducted on wastewater treated by physicochemical, activated sludge, or biofiltration processes; 1,766 E. coli isolates were obtained for the evaluation. The target disinfection level was 200 CFU/100 ml, resulting in UV and PAA doses of 7 to 30 mJ/cm(2) and 0.9 to 2.0 mg/liter, respectively. The proportions of UPECs were reduced in all samples after disinfection, with an average reduction by UV of 55% (range, 22% to 80%) and by PAA of 52% (range, 11% to 100%). Analysis of urovirulence genes revealed that the decline in the UPEC populations was not associated with any particular virulence factor. A positive association was found between the occurrence of urovirulence and antimicrobial resistance genes (ARGs). However, the changes in the prevalence of ARGs in potential UPECs were different following disinfection, i.e., UV appears to have had no effect, while PAA significantly reduced the ARG levels. Thus, this study showed that both UV and PAA disinfections reduced the proportion of UPECs and that PAA disinfection also reduced the proportion of antimicrobial resistance gene-carrying UPEC pathotypes in municipal wastewaters. PMID:24727265

Biswal, Basanta Kumar; Khairallah, Ramzi; Bibi, Kareem; Mazza, Alberto; Gehr, Ronald; Masson, Luke; Frigon, Dominic

2014-06-01

77

F9 Fimbriae of Uropathogenic Escherichia coli Are Expressed at Low Temperature and Recognise Gal?1-3GlcNAc-Containing Glycans  

PubMed Central

Uropathogenic Escherichia coli (UPEC) is the leading causative agent of urinary tract infections (UTI) in the developed world. Among the major virulence factors of UPEC, surface expressed adhesins mediate attachment and tissue tropism. UPEC strains typically possess a range of adhesins, with type 1 fimbriae and P fimbriae of the chaperone-usher class the best characterised. We previously identified and characterised F9 as a new chaperone-usher fimbrial type that mediates biofilm formation. However, the regulation and specific role of F9 fimbriae remained to be determined in the context of wild-type clinical UPEC strains. In this study we have assessed the distribution and genetic context of the f9 operon among diverse E. coli lineages and pathotypes and demonstrated that f9 genes are significantly more conserved in a UPEC strain collection in comparison to the well-defined E. coli reference (ECOR) collection. In the prototypic UPEC strain CFT073, the global regulator protein H-NS was identified as a transcriptional repressor of f9 gene expression at 37°C through its ability to bind directly to the f9 promoter region. F9 fimbriae expression was demonstrated at 20°C, representing the first evidence of functional F9 fimbriae expression by wild-type E. coli. Finally, glycan array analysis demonstrated that F9 fimbriae recognise and bind to terminal Gal?1-3GlcNAc structures. PMID:24671091

Wurpel, Daniël J.; Totsika, Makrina; Allsopp, Luke P.; Hartley-Tassell, Lauren E.; Day, Christopher J.; Peters, Kate M.; Sarkar, Sohinee; Ulett, Glen C.; Yang, Ji; Tiralongo, Joe; Strugnell, Richard A.; Jennings, Michael P.; Schembri, Mark A.

2014-01-01

78

In vivo mRNA profiling of uropathogenic Escherichia coli from diverse phylogroups reveals common and group-specific gene expression profiles.  

PubMed

mRNA profiling of pathogens during the course of human infections gives detailed information on the expression levels of relevant genes that drive pathogenicity and adaptation and at the same time allows for the delineation of phylogenetic relatedness of pathogens that cause specific diseases. In this study, we used mRNA sequencing to acquire information on the expression of Escherichia coli pathogenicity genes during urinary tract infections (UTI) in humans and to assign the UTI-associated E. coli isolates to different phylogenetic groups. Whereas the in vivo gene expression profiles of the majority of genes were conserved among 21 E. coli strains in the urine of elderly patients suffering from an acute UTI, the specific gene expression profiles of the flexible genomes was diverse and reflected phylogenetic relationships. Furthermore, genes transcribed in vivo relative to laboratory media included well-described virulence factors, small regulatory RNAs, as well as genes not previously linked to bacterial virulence. Knowledge on relevant transcriptional responses that drive pathogenicity and adaptation of isolates to the human host might lead to the introduction of a virulence typing strategy into clinical microbiology, potentially facilitating management and prevention of the disease. Importance: Urinary tract infections (UTI) are very common; at least half of all women experience UTI, most of which are caused by pathogenic Escherichia coli strains. In this study, we applied massive parallel cDNA sequencing (RNA-seq) to provide unbiased, deep, and accurate insight into the nature and the dimension of the uropathogenic E. coli gene expression profile during an acute UTI within the human host. This work was undertaken to identify key players in physiological adaptation processes and, hence, potential targets for new infection prevention and therapy interventions specifically aimed at sabotaging bacterial adaptation to the human host. PMID:25096872

Bielecki, Piotr; Muthukumarasamy, Uthayakumar; Eckweiler, Denitsa; Bielecka, Agata; Pohl, Sarah; Schanz, Ansgar; Niemeyer, Ute; Oumeraci, Tonio; von Neuhoff, Nils; Ghigo, Jean-Marc; Häussler, Susanne

2014-01-01

79

The type 1 pili regulator gene fimX and pathogenicity island PAI-X as molecular markers of uropathogenic Escherichia coli  

PubMed Central

Uropathogenic Escherichia coli (UPEC) fall within a larger group of isolates producing extraintestinal disease. UPEC express type 1 pili as a critical virulence determinant mediating adherence to and invasion into urinary tract tissues. Type 1 pili expression is under regulation by a family of site-specific recombinases, including FimX, which is encoded from a genomic island called PAI-X for pathogenicity island of FimX. Using a new multiplex PCR, fimX and the additional PAI-X genes were found to be highly associated with UPEC (144/173?=?83.2?%), and more prevalent in UPEC of lower urinary tract origin (105/120?=?87.5?%) than upper urinary tract origin (39/53?=?74?%; P<0.05) or commensal isolates (28/78?=?36?%; P?0.0001). The Fim-like recombinase gene fimX is the only family member that has a significant association with UPEC compared to commensal isolates. Our results indicate PAI-X genes, including the type 1 pili regulator gene fimX, are highly prevalent among UPEC isolates and have a strong positive correlation with genomic virulence factors, suggesting a potential role for PAI-X in the extraintestinal pathogenic E. coli lifestyle. PMID:23744903

Bateman, Stacey L.; Stapleton, Ann E.; Stamm, Walter E.; Hooton, Thomas M.

2013-01-01

80

A unique arabinose 5-phosphate isomerase found within a genomic island associated with the uropathogenicity of Escherichia coli CFT073.  

PubMed

Previous studies showed that deletion of genes c3405 to c3410 from PAI-metV, a genomic island from Escherichia coli CFT073, results in a strain that fails to compete with wild-type CFT073 after a transurethral cochallenge in mice and is deficient in the ability to independently colonize the mouse kidney. Our analysis of c3405 to c3410 suggests that these genes constitute an operon with a role in the internalization and utilization of an unknown carbohydrate. This operon is not found in E. coli K-12 but is present in a small number of pathogenic E. coli and Shigella boydii strains. One of the genes, c3406, encodes a protein with significant homology to the sugar isomerase domain of arabinose 5-phosphate isomerases but lacking the tandem cystathionine beta-synthase domains found in the other arabinose 5-phosphate isomerases of E. coli. We prepared recombinant c3406 protein, found it to possess arabinose 5-phosphate isomerase activity, and characterized this activity in detail. We also constructed a c3406 deletion mutant of E. coli CFT073 and demonstrated that this deletion mutant was still able to compete with wild-type CFT073 in a transurethral cochallenge in mice and could colonize the mouse kidney. These results demonstrate that the presence of c3406 is not essential for a pathogenic phenotype. PMID:21498648

Mosberg, Joshua A; Yep, Alejandra; Meredith, Timothy C; Smith, Sara; Wang, Pan-Fen; Holler, Tod P; Mobley, Harry L T; Woodard, Ronald W

2011-06-01

81

In vitro potency and efficacy favor later generation fluoroquinolones for treatment of canine and feline Escherichia coli uropathogens in the United States.  

PubMed

Information regarding in vitro activity of newer fluoroquinolones (FQs) is limited despite increasing resistance in canine or feline pathogenic Escherichia coli (E. coli). This study describes in vitro potency and efficacy toward E. coli of seven FQs grouped according to similarities in chemical structure: enrofloxacin, ciprofloxacin, orbifloxacin (first-group), levofloxacin, marbofloxacin (second-group) and pradofloxacin, moxifloxacin (third-group; latest S, S-pyrrolidino-piperidine at C-7). Potency measures included minimum inhibitory concentration (MIC) (geometric mean MIC, MIC(50), MIC(90)); and mutant prevention concentration (MPC) for FQ susceptible isolates only. In vitro efficacy measures included relative susceptibility (MIC(BP-S):MIC) or resistance (MIC:MIC(BP-R)) and mutant selection window (MSW) (MPC:MIC). For enrofloxacin susceptible isolates, mean MIC (?g/ml) was least for each third-group drug and ciprofloxacin and greatest for enrofloxacin and orbifloxacin (P = 0.006). For enrofloxacin susceptible isolates, MPC were below MIC:MIC(BP-R) and least for pradofloxacin (0.29 ± 0.16 ?g/ml) and greatest for enrofloxacin (1.55 ± 0.55 ?g/ml) (P = 0.006). MSW was least for pradofloxacin (55 ± 30) and greatest for ciprofloxacin (152 ± 76) (P = 0.0024). MIC(BP-S):MIC was greatest (P = 0.025) for pradofloxacin (190.1 ± 0.61) and least for enrofloxacin (23.53 ± 0.83). For FQ susceptible isolates, FQs MIC:MIC(BP-R) may serve as a surrogate for MPC. Because in vitro efficacy was greatest for pradofloxacin; it might be preferred for treatment of urinary tract infections (UTIs) associated with FQ susceptible E. coli uropathogens. PMID:23136054

Liu, Xiaoqiang; Boothe, Dawn M; Jin, Yaping; Thungrat, Kamoltip

2013-02-01

82

Inhibitors of TonB Function Identified by a High-Throughput Screen for Inhibitors of Iron Acquisition in Uropathogenic Escherichia coli CFT073  

PubMed Central

ABSTRACT The urinary tract is one of the most common sites of infection in humans, and uropathogenic Escherichia coli (UPEC) is the main causative agent of urinary tract infections. Bacteria colonizing the urinary tract face extremely low iron availability. To counteract this, UPEC expresses a wide variety of iron acquisition systems. To exploit iron acquisition in UPEC as a global target for small-molecule inhibition, we developed and carried out a whole-cell growth-based high throughput screen of 149,243 compounds. Our primary assay was carried out under iron-limiting conditions. Hits in the primary screen were assayed using two counterscreens that ruled out iron chelators and compounds that inhibit growth by means other than inhibition of iron acquisition. We determined dose-response curves under two different iron conditions and purchased fresh compounds for selected hits. After retesting dose-response relationships, we identified 16 compounds that arrest growth of UPEC only under iron-limiting conditions. All compounds are bacteriostatic and do not inhibit proton motive force. A loss-of-target strategy was employed to identify the cellular target of these inhibitors. Two compounds lost inhibitory activity against a strain lacking TonB and were shown to inhibit irreversible adsorption of a TonB-dependent bacteriophage. Our results validate iron acquisition as a target for antibacterial strategies against UPEC and identify TonB as one of the cellular targets. PMID:24570372

Yep, Alejandra; McQuade, Thomas; Kirchhoff, Paul; Larsen, Martha; Mobley, Harry L. T.

2014-01-01

83

Intranasal immunization with fusion protein MrpH·FimH and MPL adjuvant confers protection against urinary tract infections caused by uropathogenic Escherichia coli and Proteus mirabilis.  

PubMed

Urinary tract infections (UTIs) caused by Uropathogenic Escherichia coli (UPEC) and Proteus mirabilis are among the most common infections in the world. Currently there are no vaccines available to confer protection against UTI in humans. In this study, the immune responses and protection of FimH of UPEC with MrpH antigen of P. mirabilis in different vaccine formulations with and without MPL adjuvant were assessed. Mice intranasally immunized with the novel fusion protein MrpH·FimH induced a significant increase in IgG and IgA in serum, nasal wash, vaginal wash, and urine samples. Mice immunized with fusion MrpH·FimH also showed a significant boost in cellular immunity. Addition of MPL as the adjuvant enhanced FimH and MrpH specific humoral and cellular responses in both systemic and mucosal samples. Vaccination with MrpH·FimH alone or in combination with MPL showed the highest efficiency in clearing bladder and kidney infections in mice challenged with UPEC and P. mirabilis. These findings may indicate that the protection observed correlates with the systemic, mucosal and cellular immune responses induced by vaccination with these preparations. Our data suggest MrpH·FimH fusion protein with or without MPL as adjuvant could be potential vaccine candidates for elimination of UPEC and P. mirabilis. These data altogether are promising and these formulations are good candidates for elimination of UPEC and P. mirabilis. PMID:25562574

Habibi, Mehri; Asadi Karam, Mohammad Reza; Shokrgozar, Mohammad Ali; Oloomi, Mana; Jafari, Anis; Bouzari, Saeid

2015-04-01

84

A-Type proanthocyanidin trimers from cranberry that inhibit adherence of uropathogenic P-fimbriated Escherichia coli.  

PubMed

Three proanthocyanidin trimers possessing A-type interflavanoid linkages, epicatechin-(4beta-->6)-epicatechin-(4beta-->8, 2beta-->O-->7)-epicatechin (4), epicatechin-(4beta-->8, 2beta-->O-->7)-epicatechin-(4beta-->8)-epicatechin (5), and epicatechin-(4beta-->8)-epicatechin-(4beta-->8, 2beta-->O-->7)-epicatechin (6), were isolated from the ripe fruits of Vaccinium macrocarpon (cranberry) and prevented adherence of P-fimbriated Escherichia coli isolates from the urinary tract to cellular surfaces containing alpha-Gal(1-->4)beta-Gal receptor sequences similar to those on uroepithelial cells. The structure of 4 was elucidated by a combination of spectroscopic methods and acid-catalyzed degradation with phloroglucinol. Also isolated were the weakly active epicatechin-(4beta-->8, 2beta-->O-->7)-epicatechin (procyanidin A2) (3) and the inactive monomer epicatechin (1) and the inactive dimer epicatechin-(4beta-->8)-epicatechin (procyanidin B2) (2). PMID:11000024

Foo, L Y; Lu, Y; Howell, A B; Vorsa, N

2000-09-01

85

Quantitative Analysis of Amyloid-Integrated Biofilms Formed by Uropathogenic Escherichia coli at the Air-Liquid Interface  

PubMed Central

Bacterial biofilms are complex multicellular assemblies, characterized by a heterogeneous extracellular polymeric matrix, that have emerged as hallmarks of persistent infectious diseases. New approaches and quantitative data are needed to elucidate the composition and architecture of biofilms, and such data need to be correlated with mechanical and physicochemical properties that relate to function. We performed a panel of interfacial rheological measurements during biofilm formation at the air-liquid interface by the Escherichia coli strain UTI89, which is noted for its importance in studies of urinary tract infection and for its assembly of functional amyloid fibers termed curli. Brewster-angle microscopy and measurements of the surface elasticity (Gs?) and stress-strain response provided sensitive and quantitative parameters that revealed distinct stages during bacterial colonization, aggregation, and eventual formation of a pellicle at the air-liquid interface. Pellicles that formed under conditions that upregulate curli production exhibited an increase in strength and viscoelastic properties as well as a greater ability to recover from stress-strain perturbation. The results suggest that curli, as hydrophobic extracellular amyloid fibers, enhance the strength, viscoelasticity, and resistance to strain of E. coli biofilms formed at the air-liquid interface. PMID:22947862

Wu, Cynthia; Lim, Ji Youn; Fuller, Gerald G.; Cegelski, Lynette

2012-01-01

86

Distribution of drb genes coding for Dr binding adhesins among uropathogenic and fecal Escherichia coli isolates and identification of new subtypes.  

PubMed Central

The Dr family of related adherence structures, some fimbriated and others afimbriated, bind to decay-accelerating factor molecules on human cells. Dr is associated with recurring urinary tract infection (UTI), but the distribution of Dr subtypes among uropathogenic Escherichia coli causing UTI among otherwise healthy women has yet to be described. A total of 787 UTI and fecal E. coli isolates from college women were screened for the presence of Dr sequences (drb). Fifteen percent of UTI strains were drb positive, compared to 5% of fecal strains. The adhesin (E gene) subtype of each drb-positive strain was determined by type-specific PCR followed by restriction enzyme analysis. Among 78 drb-positive strains, we found 14 (18%) afaE1, 1 (1.3%) afaE2, 1 (1.3%) afaE3, 9 (12%) draE, 9 (12%) draE-afaE3 hybrid, 1 (1.3%) daaE, 32 (41%) afaE5, 4 (5.1%) F131 E gene-like, and 7 untypeable strains. All untypeable E genes were cloned and sequenced, revealing four additional new classes of E genes, including two similar to the previously identified nonfimbrial E series. While a great range of diversity exists among the E genes, restriction fragment length polymorphism analysis demonstrated that all of these drb operons share a highly conserved gene structure. The most common subtype, afaE5, occurred three times as often among UTI than fecal strains. Over half of the drb-positive strains and 80% of those positive for afaE5 have the same virulence signature (positive for aer, kpsMT, ompT, and fim), suggesting an association of this profile with UTI pathogenesis. PMID:9169726

Zhang, L; Foxman, B; Tallman, P; Cladera, E; Le Bouguenec, C; Marrs, C F

1997-01-01

87

Th1-Th17 Cells Contribute to the Development of Uropathogenic Escherichia coli-Induced Chronic Pelvic Pain  

PubMed Central

The etiology of chronic prostatitis/chronic pelvic pain syndrome in men is unknown but may involve microbes and autoimmune mechanisms. We developed an infection model of chronic pelvic pain in NOD/ShiLtJ (NOD) mice with a clinical Escherichia coli isolate (CP-1) from a patient with chronic pelvic pain. We investigated pain mechanisms in NOD mice and compared it to C57BL/6 (B6) mice, a strain resistant to CP-1-induced pain. Adoptive transfer of CD4+ T cells, but not serum, from CP-1-infected NOD mice was sufficient to induce chronic pelvic pain. CD4+ T cells in CP-1-infected NOD mice expressed IFN-? and IL-17A but not IL-4, consistent with a Th1/Th17 immune signature. Adoptive transfer of ex-vivo expanded IFN-? or IL-17A-expressing cells was sufficient to induce pelvic pain in naïve NOD recipients. Pelvic pain was not abolished in NOD-IFN-?-KO mice but was associated with an enhanced IL-17A immune response to CP1 infection. These findings demonstrate a novel role for Th1 and Th17-mediated adaptive immune mechanisms in chronic pelvic pain. PMID:23577183

Mukherjee, Soumi; Done, Joseph D.; Schaeffer, Anthony J.; Thumbikat, Praveen

2013-01-01

88

Uropathogenic Escherichia coli P and Type 1 Fimbriae Act in Synergy in a Living Host to Facilitate Renal Colonization Leading to Nephron Obstruction  

PubMed Central

The progression of a natural bacterial infection is a dynamic process influenced by the physiological characteristics of the target organ. Recent developments in live animal imaging allow for the study of the dynamic microbe-host interplay in real-time as the infection progresses within an organ of a live host. Here we used multiphoton microscopy-based live animal imaging, combined with advanced surgical procedures, to investigate the role of uropathogenic Escherichia coli (UPEC) attachment organelles P and Type 1 fimbriae in renal bacterial infection. A GFP+ expressing variant of UPEC strain CFT073 and genetically well-defined isogenic mutants were microinfused into rat glomerulus or proximal tubules. Within 2 h bacteria colonized along the flat squamous epithelium of the Bowman's capsule despite being exposed to the primary filtrate. When facing the challenge of the filtrate flow in the proximal tubule, the P and Type 1 fimbriae appeared to act in synergy to promote colonization. P fimbriae enhanced early colonization of the tubular epithelium, while Type 1 fimbriae mediated colonization of the center of the tubule via a mechanism believed to involve inter-bacterial binding and biofilm formation. The heterogeneous bacterial community within the tubule subsequently affected renal filtration leading to total obstruction of the nephron within 8 h. Our results reveal the importance of physiological factors such as filtration in determining bacterial colonization patterns, and demonstrate that the spatial resolution of an infectious niche can be as small as the center, or periphery, of a tubule lumen. Furthermore, our data show how secondary physiological injuries such as obstruction contribute to the full pathophysiology of pyelonephritis. PMID:21383970

Melican, Keira; Sandoval, Ruben M.; Kader, Abdul; Josefsson, Lina; Tanner, George A.; Molitoris, Bruce A.; Richter-Dahlfors, Agneta

2011-01-01

89

Escherichia Coli  

ERIC Educational Resources Information Center

Diverse biological data may be used to create illustrations of molecules in their cellular context. I describe the scientific results that support a recent textbook illustration of an "Escherichia coli cell". The image magnifies a portion of the bacterium at one million times, showing the location and form of individual macromolecules. Results…

Goodsell, David S.

2009-01-01

90

Mobilisation and remobilisation of a large archetypal pathogenicity island of uropathogenic Escherichia coli in vitro support the role of conjugation for horizontal transfer of genomic islands  

PubMed Central

Background A substantial amount of data has been accumulated supporting the important role of genomic islands (GEIs) - including pathogenicity islands (PAIs) - in bacterial genome plasticity and the evolution of bacterial pathogens. Their instability and the high level sequence similarity of different (partial) islands suggest an exchange of PAIs between strains of the same or even different bacterial species by horizontal gene transfer (HGT). Transfer events of archetypal large genomic islands of enterobacteria which often lack genes required for mobilisation or transfer have been rarely investigated so far. Results To study mobilisation of such large genomic regions in prototypic uropathogenic E. coli (UPEC) strain 536, PAI II536 was supplemented with the mobRP4 region, an origin of replication (oriVR6K), an origin of transfer (oriTRP4) and a chloramphenicol resistance selection marker. In the presence of helper plasmid RP4, conjugative transfer of the 107-kb PAI II536 construct occured from strain 536 into an E. coli K-12 recipient. In transconjugants, PAI II536 existed either as a cytoplasmic circular intermediate (CI) or integrated site-specifically into the recipient's chromosome at the leuX tRNA gene. This locus is the chromosomal integration site of PAI II536 in UPEC strain 536. From the E. coli K-12 recipient, the chromosomal PAI II536 construct as well as the CIs could be successfully remobilised and inserted into leuX in a PAI II536 deletion mutant of E. coli 536. Conclusions Our results corroborate that mobilisation and conjugal transfer may contribute to evolution of bacterial pathogens through horizontal transfer of large chromosomal regions such as PAIs. Stabilisation of these mobile genetic elements in the bacterial chromosome result from selective loss of mobilisation and transfer functions of genomic islands. PMID:21943043

2011-01-01

91

Correlation between uropathogenic properties of Escherichia coli from urinary tract infections and the antibody-coated bacteria test and comparison with faecal strains.  

PubMed

Strains of Escherichia coli isolated from adult females with symptomatic urinary tract infection were found to possess the following properties significantly more frequently than faecal strains: (i) high K-antigen titre: (ii) haemolysin; (iii) type 1 pili; (iv) mannose-resistant haemagglutination; (v) fermentation of dulcitol and salicin; (vi) O serotype 2, 6 and 75; (vii) H serotype 1. E. coli isolated form urine specimens containing significant numbers of antibody-coated bacteria were richer in these seven properties than strains from urines without detectable antibody coated bacteria. The O and H serotypes of E. coli obtained from patients with urinary tract infection in two New Zealand cities were compared with those reported in the world literature and found to be similar. PMID:6114119

Brooks, H J; Benseman, B A; Peck, J; Bettelheim, K A

1981-08-01

92

[Prevalence of beta-lactamase CTX-M-15 in phylogenetic groups of uropathogenic Escherichia coli isolated from patients in the community of Merida, Venezuela].  

PubMed

In this study we determined the prevalence of extended-spectrum beta-lactamases (ESBLs) in phylogenetic groups of uropathogenic E. coli (UPEC) isolated from patients in the community. Twenty one UPEC strains with reduced susceptibility to broad-spectrum cephalosporins were collected between January 2009 and July 2010, from patients with urinary tract infection who attended the Public Health Laboratory in Mérida, Venezuela. Genotypic characterization determined that all UPEC strains harbored blaBLEEs genes: 76.2% of the strains showed the presence of a single ESBL-producer gene, represented by blaCTX-M-15, whereas 23.8% of UPEC showed various combinations of bla genes (blacCTX-M-15 + blaTEM-1, blaCTX-M-15 + blaSHV and blaSHV + blaTEM-1). In this study, 61.9% of the isolates were placed in phylogroup A and the remaining strains were assigned to group B2 (38.1%). There was no evidence of spread of a particular UPEC clone; only seven strains belonged to a clonal group with an index of similarity greater than 85%. To our knowledge, this is the first description of blxCTX-M-15 in UPEC from patients with community-acquired urinary tract infections, which shows that Venezuela is also part of the so-called CTX-M-15 pandemic. The findings in this study, as well as its clinical and epidemiological implications, lead to the need for monitoring and controlling the spread of CTX-M-15 producing UPECs, not only regionally, but also nationwide. PMID:24758100

Hernández, Erick; Araque, María; Millán, Ysheth; Millán, Beatriz; Vielma, Silvana

2014-03-01

93

Characteristics and prevalence within serogroup O4 of a J96-like clonal group of uropathogenic Escherichia coli O4:H5 containing the class I and class III alleles of papG.  

PubMed

The recent discovery of a geographically dispersed clonal group of Escherichia coli O4:H5 that includes prototypic uropathogenic strain J96 prompted us to determine the prevalence of J96-like strains within serogroup O4 and to further assess the characteristics of such strains. We used O:K:H;F serotyping, PCR-based genomic fingerprinting, pulsed-field gel electrophoresis (PFGE), multilocus enzyme electrophoresis (MLEE), and PCR detection of the three papG alleles and of the cytotoxic necrotizing factor 1 (cnf1) and aerobactin (aer) gene sequences to characterize the 15 O4 strains among 336 E. coli isolates from three clinical collections (187 from mixed-source bacteremia, 75 from urosepsis, and 74 from acute cystitis). J96-like strains constituted approximately half of the O4 strains, or 2% of the total population. In contrast to other O4 strains, the J96-like strains characteristically exhibited specific group III capsular antigens, the H5 flagellar and F13 fimbrial antigens, a distinctive PCR genomic fingerprint, the class III papG allele (plus, in 50% of strains, the enigmatic class I papG allele), and cnf1 but lacked aer. A subset of these strains was remarkably homogeneous with respect to all these characteristics and exhibited a distinctive PFGE fingerprint and MLEE pattern. These findings clarify the epidemiological relevance of J96 as a model extraintestinal pathogen, provide further evidence of the class I papG allele outside of strain J96, and offer insights into the evolution of E. coli serogroup O4. PMID:9169745

Johnson, J R; Stapleton, A E; Russo, T A; Scheutz, F; Brown, J J; Maslow, J N

1997-06-01

94

In Vivo mRNA Profiling of Uropathogenic Escherichia coli from Diverse Phylogroups Reveals Common and Group-Specific Gene Expression Profiles  

PubMed Central

ABSTRACT mRNA profiling of pathogens during the course of human infections gives detailed information on the expression levels of relevant genes that drive pathogenicity and adaptation and at the same time allows for the delineation of phylogenetic relatedness of pathogens that cause specific diseases. In this study, we used mRNA sequencing to acquire information on the expression of Escherichia coli pathogenicity genes during urinary tract infections (UTI) in humans and to assign the UTI-associated E. coli isolates to different phylogenetic groups. Whereas the in vivo gene expression profiles of the majority of genes were conserved among 21 E. coli strains in the urine of elderly patients suffering from an acute UTI, the specific gene expression profiles of the flexible genomes was diverse and reflected phylogenetic relationships. Furthermore, genes transcribed in vivo relative to laboratory media included well-described virulence factors, small regulatory RNAs, as well as genes not previously linked to bacterial virulence. Knowledge on relevant transcriptional responses that drive pathogenicity and adaptation of isolates to the human host might lead to the introduction of a virulence typing strategy into clinical microbiology, potentially facilitating management and prevention of the disease. PMID:25096872

Bielecki, Piotr; Muthukumarasamy, Uthayakumar; Eckweiler, Denitsa; Bielecka, Agata; Pohl, Sarah; Schanz, Ansgar; Niemeyer, Ute; Oumeraci, Tonio; von Neuhoff, Nils; Ghigo, Jean-Marc

2014-01-01

95

Pyelonephritogenic Diffusely Adhering Escherichia coli EC7372 Harboring Dr-II Adhesin Carries Classical Uropathogenic Virulence Genes and Promotes Cell Lysis and Apoptosis in Polarized Epithelial Caco-2/TC7 Cells  

PubMed Central

Diffusely adhering Escherichia coli (DAEC) strains expressing adhesins of the Afa/Dr family bind to epithelial cells in a diffuse adherence pattern by recognizing a common receptor, the decay-accelerating factor (CD55). Recently, a novel CD55-binding adhesin, named Dr-II, was identified from the pyelonephritogenic strain EC7372. In this report, we show that despite the low level of sequence identity between Dr-II and other members of the Afa/Dr family, EC7372 induces pathophysiological effects similar to those induced by other Afa/Dr DAEC strains on the polarized epithelial cell line Caco-2/TC7. Specifically, the Dr-II adhesin was sufficient to promote CD55 and CD66e clustering around adhering bacteria and apical cytoskeleton rearrangements. Unlike other Afa/Dr DAEC strains, EC7372 expresses a functional hemolysin that promotes a rapid cellular lysis. In addition, cell death by apoptosis or necrosis was observed in EC7372-infected Caco-2/TC7 cells, depending on infection time. Our results indicate that EC7372 harbors a pathogenicity island (PAI) similar to the one described for the pyelonephritogenic strain CFT073, which carries both hly and pap operons. Cumulatively, our findings indicate that strain EC7372 can be considered a prototype of a subclass of Afa/Dr DAEC isolates that have acquired a PAI harboring several classical uropathogenic virulence genes. PMID:11083827

Guignot, Julie; Breard, Jacqueline; Bernet-Camard, Marie-Françoise; Peiffer, Isabelle; Nowicki, Bogdan J.; Servin, Alain L.; Blanc-Potard, Anne-Beatrice

2000-01-01

96

Propolis can potentialise the anti-adhesion activity of proanthocyanidins on uropathogenic Escherichia coli in the prevention of recurrent urinary tract infections  

PubMed Central

Background Escherichia coli, the main bacteria found in recurrent urinary tract infections (UTI), is now frequently resistant to several currently used antibiotic treatments making new solutions essential. In this study, we evaluated the association propolis and proanthocyanidins type A to reduce bacterial anti-adhesion activity of E. coli on urothelial cells. Results This first double-blind, randomized, cross-over human trial included 5 volunteers that followed 6 different regimens with or without variable doses of cranberry and propolis with a washout period of at least 1 week between each regimen. Urine samples were collected at 0 h, 4-6 h, 12 h and 24 h after cranberry plus propolis or placebo capsule consumption. In vivo urinary bacterial anti-adhesion activity was assessed with a bioassay (a human T24 epithelial cell-line assay) and an in vivo Caenorhabditis elegans model. HPLC-PDA-MS was used to detect propolis and cranberry compounds in urine. Bioassays indicated significant bacterial anti-adhesion activity in urine collected from volunteers who had consumed cranberry plus propolis powder compared to placebo (p < 0.001). This inhibition was clearly dose-dependent, increasing with the amount of PACs and propolis equivalents consumed in each regimen. Results suggested that propolis had an additional effect with PACs and prevent a bacterial anti-adhesion effect over 1 day. An in vivo model showed that the E. coli strain presented a reduced ability to kill C. elegans after their growth in urine samples of patients who took cranberry plus propolis capsules. HPLC confirmed that propolis is excreted in urine. Conclusions This study presents an alternative to prevent recurrent UTI. Administration of PACs plus propolis once daily offers some protection against bacterial adhesion, bacterial multiplication and virulence in the urinary tract, representing an interesting new strategy to prevent recurrent UTI. PMID:22126300

2011-01-01

97

Multiresistant Uropathogenic Escherichia coli from a Region in India Where Urinary Tract Infections Are Endemic: Genotypic and Phenotypic Characteristics of Sequence Type 131 Isolates of the CTX-M-15 Extended-Spectrum-?-Lactamase-Producing Lineage  

PubMed Central

Escherichia coli sequence type 131 (O25b:H4), associated with the CTX-M-15 extended-spectrum beta-lactamases (ESBLs) and linked predominantly to the community-onset antimicrobial-resistant infections, has globally emerged as a public health concern. However, scant attention is given to the understanding of the molecular epidemiology of these strains in high-burden countries such as India. Of the 100 clinical E. coli isolates obtained by us from a setting where urinary tract infections are endemic, 16 ST131 E. coli isolates were identified by multilocus sequence typing (MLST). Further, genotyping and phenotyping methods were employed to characterize their virulence and drug resistance patterns. All the 16 ST131 isolates harbored the CTX-M-15 gene, and half of them also carried TEM-1; 11 of these were positive for blaOXA groups 1 and 12 for aac(6?)-Ib-cr. At least 12 isolates were refractory to four non-beta-lactam antibiotics: ciprofloxacin, gentamicin, sulfamethoxazole-trimethoprim, and tetracycline. Nine isolates carried the class 1 integron. Plasmid analysis indicated a large pool of up to six plasmids per strain with a mean of approximately three plasmids. Conjugation and PCR-based replicon typing (PBRT) revealed that the spread of resistance was associated with the FIA incompatibility group of plasmids. Pulsed-field gel electrophoresis (PFGE) and genotyping of the virulence genes showed a low level of diversity among these strains. The association of ESBL-encoding plasmid with virulence was demonstrated in transconjugants by serum assay. None of the 16 ST131 ESBL-producing E. coli strains were known to synthesize carbapenemase enzymes. In conclusion, our study reports a snapshot of the highly virulent/multiresistant clone ST131 of uropathogenic E. coli from India. This study suggests that the ST131 genotypes from this region are clonally evolved and are strongly associated with the CTX-M-15 enzyme, carry a high antibiotic resistance background, and have emerged as an important cause of community-acquired urinary tract infections. PMID:23045357

Hussain, Arif; Ewers, Christa; Nandanwar, Nishant; Guenther, Sebastian; Jadhav, Savita; Wieler, Lothar H.

2012-01-01

98

Multiresistant uropathogenic Escherichia coli from a region in India where urinary tract infections are endemic: genotypic and phenotypic characteristics of sequence type 131 isolates of the CTX-M-15 extended-spectrum-?-lactamase-producing lineage.  

PubMed

Escherichia coli sequence type 131 (O25b:H4), associated with the CTX-M-15 extended-spectrum beta-lactamases (ESBLs) and linked predominantly to the community-onset antimicrobial-resistant infections, has globally emerged as a public health concern. However, scant attention is given to the understanding of the molecular epidemiology of these strains in high-burden countries such as India. Of the 100 clinical E. coli isolates obtained by us from a setting where urinary tract infections are endemic, 16 ST131 E. coli isolates were identified by multilocus sequence typing (MLST). Further, genotyping and phenotyping methods were employed to characterize their virulence and drug resistance patterns. All the 16 ST131 isolates harbored the CTX-M-15 gene, and half of them also carried TEM-1; 11 of these were positive for bla(OXA) groups 1 and 12 for aac(6')-Ib-cr. At least 12 isolates were refractory to four non-beta-lactam antibiotics: ciprofloxacin, gentamicin, sulfamethoxazole-trimethoprim, and tetracycline. Nine isolates carried the class 1 integron. Plasmid analysis indicated a large pool of up to six plasmids per strain with a mean of approximately three plasmids. Conjugation and PCR-based replicon typing (PBRT) revealed that the spread of resistance was associated with the FIA incompatibility group of plasmids. Pulsed-field gel electrophoresis (PFGE) and genotyping of the virulence genes showed a low level of diversity among these strains. The association of ESBL-encoding plasmid with virulence was demonstrated in transconjugants by serum assay. None of the 16 ST131 ESBL-producing E. coli strains were known to synthesize carbapenemase enzymes. In conclusion, our study reports a snapshot of the highly virulent/multiresistant clone ST131 of uropathogenic E. coli from India. This study suggests that the ST131 genotypes from this region are clonally evolved and are strongly associated with the CTX-M-15 enzyme, carry a high antibiotic resistance background, and have emerged as an important cause of community-acquired urinary tract infections. PMID:23045357

Hussain, Arif; Ewers, Christa; Nandanwar, Nishant; Guenther, Sebastian; Jadhav, Savita; Wieler, Lothar H; Ahmed, Niyaz

2012-12-01

99

Lactuca indica extract interferes with uroepithelial infection by Escherichia coli  

Microsoft Academic Search

Ethnopharmacological relevanceUropathogenic Escherichia coli is the major cause for urinary tract infections (UTI). Due to emerging antimicrobial resistances treatment of UTI is becoming increasingly difficult. Therefore, alternative treatment strategies are required. We sought to investigate the molecular mechanisms of a traditionally used decoction from Vietnamese dandelion (Lactuca indica L.) mediating local protection of the bladder epithelium.

Petra Lüthje; Dang Ngoc Dzung; Annelie Brauner

2011-01-01

100

Target analysis of ?-alkylidene-?-butyrolactones in uropathogenic E. coli.  

PubMed

?-Alkylidene-?-butyrolactones are quite common in nature and exhibit a broad spectrum of biological activities. We therefore synthesized a small library of xanthatine inspired ?-alkylidene-?-butyrolactones to screen non-pathogenic and uropathogenic E. coli strains by activity based protein profiling (ABPP). The identified targets are involved in cellular redox processes and give first insight into the preferred binding sites of this privileged motif. Furthermore the gene of one protein, c2450, which was only identified in uropathogenic E. coli belongs to a genomic island which encodes a hybrid polyketide/non-ribosomal peptide synthetase (PKS/NRPS). This system is responsible for the synthesis of colibactin, a natural product which causes DNA double strand breaks in eukaryotic cells leading to the activation of the DNA damage checkpoint pathway and subsequent cell cycle arrest. While the role of several proteins that are involved in the colibactin synthesis has been elucidated, the function of c2450 remains elusive. Investigation of the binding site showed that c2450 is modified at a cysteine residue which may be important for the catalytic activity. PMID:22990910

Kunzmann, Martin H; Sieber, Stephan A

2012-11-01

101

Dosage effect on uropathogenic Escherichia coli anti-adhesion activity in urine following consumption of cranberry powder standardized for proanthocyanidin content: a multicentric randomized double blind study  

Microsoft Academic Search

BACKGROUND: Ingestion of cranberry (Vaccinium macrocarpon Ait.) has traditionally been utilized for prevention of urinary tract infections. The proanthocyanidins (PACs) in cranberry, in particular the A-type linkages have been implicated as important inhibitors of primarily P-fimbriated E. coli adhesion to uroepithelial cells. Additional experiments were required to investigate the persistence in urine samples over a broader time period, to determine

Amy B Howell; Henry Botto; Christophe Combescure; Anne-Béatrice Blanc-Potard; Lluis Gausa; Tetsuro Matsumoto; Peter Tenke; Albert Sotto; Jean-Philippe Lavigne

2010-01-01

102

Dosage effect on uropathogenic Escherichia coli anti-adhesion activity in urine following consumption of cranberry powder standardized for proanthocyanidin content: a multicentric randomized double blind study  

PubMed Central

Background Ingestion of cranberry (Vaccinium macrocarpon Ait.) has traditionally been utilized for prevention of urinary tract infections. The proanthocyanidins (PACs) in cranberry, in particular the A-type linkages have been implicated as important inhibitors of primarily P-fimbriated E. coli adhesion to uroepithelial cells. Additional experiments were required to investigate the persistence in urine samples over a broader time period, to determine the most effective dose per day and to determine if the urinary anti-adhesion effect following cranberry is detected within volunteers of different origins. Methods Two separate bioassays (a mannose-resistant hemagglutination assay and an original new human T24 epithelial cell-line assay) have assessed the ex-vivo urinary bacterial anti-adhesion activity on urines samples collected from 32 volunteers from Japan, Hungary, Spain and France in a randomized, double-blind versus placebo study. An in vivo Caenorhabditis elegans model was used to evaluate the influence of cranberry regimen on the virulence of E. coli strain. Results The results indicated a significant bacterial anti-adhesion activity in urine samples collected from volunteers that consumed cranberry powder compared to placebo (p < 0.001). This inhibition was clearly dose-dependent, prolonged (until 24 h with 72 mg of PAC) and increasing with the amount of PAC equivalents consumed in each cranberry powder regimen. An in vivo Caenorhabditis elegans model showed that cranberry acted against bacterial virulence: E. coli strain presented a reduced ability to kill worms after a growth in urines samples of patients who took cranberry capsules. This effect is particularly important with the regimen of 72 mg of PAC. Conclusions Administration of PAC-standardized cranberry powder at dosages containing 72 mg of PAC per day may offer some protection against bacterial adhesion and virulence in the urinary tract. This effect may offer a nyctohemeral protection. PMID:20398248

2010-01-01

103

The Genome Sequence of Avian Pathogenic Escherichia coli Strain O1:K1:H7 Shares Strong Similarities with Human Extraintestinal Pathogenic E. coli Genomes  

Microsoft Academic Search

Escherichia coli strains that cause disease outside the intestine are known as extraintestinal pathogenic E. coli (ExPEC) and include human uropathogenic E. coli (UPEC) and avian pathogenic E. coli (APEC). Regard- less of host of origin, ExPEC strains share many traits. It has been suggested that these commonalities may enable APEC to cause disease in humans. Here, we begin to

Timothy J. Johnson; Subhashinie Kariyawasam; Yvonne Wannemuehler; Paul Mangiamele; Sara J. Johnson; Curt Doetkott; Jerod A. Skyberg; Aaron M. Lynne; James R. Johnson; Lisa K. Nolan

2007-01-01

104

Diarrheagenic Escherichia coli  

PubMed Central

Escherichia coli is the predominant nonpathogenic facultative flora of the human intestine. Some E. coli strains, however, have developed the ability to cause disease of the gastrointestinal, urinary, or central nervous system in even the most robust human hosts. Diarrheagenic strains of E. coli can be divided into at least six different categories with corresponding distinct pathogenic schemes. Taken together, these organisms probably represent the most common cause of pediatric diarrhea worldwide. Several distinct clinical syndromes accompany infection with diarrheagenic E. coli categories, including traveler’s diarrhea (enterotoxigenic E. coli), hemorrhagic colitis and hemolytic-uremic syndrome (enterohemorrhagic E. coli), persistent diarrhea (enteroaggregative E. coli), and watery diarrhea of infants (enteropathogenic E. coli). This review discusses the current level of understanding of the pathogenesis of the diarrheagenic E. coli strains and describes how their pathogenic schemes underlie the clinical manifestations, diagnostic approach, and epidemiologic investigation of these important pathogens. PMID:9457432

Nataro, James P.; Kaper, James B.

1998-01-01

105

Pathogenic Escherichia coli  

Technology Transfer Automated Retrieval System (TEKTRAN)

Escherichia coli, a member of the Enterobacteriaceae family, is a part of the normal flora of the intestinal tract of humans and a variety of animals. E. coli strains are classified on the basis of antigenic differences in two surface components (serotyping), the somatic antigen (O) of the lipopoly...

106

Role of Enteroaggregative Escherichia coli Virulence Factors in Uropathogenesis  

PubMed Central

A multiresistant clonal Escherichia coli O78:H10 strain qualifying molecularly as enteroaggregative Escherichia coli (EAEC) was recently shown to be the cause of a community-acquired outbreak of urinary tract infection (UTI) in greater Copenhagen, Denmark, in 1991. This marks the first time EAEC has been associated with an extraintestinal disease outbreak. Importantly, the outbreak isolates were recovered from the urine of patients with symptomatic UTI, strongly implying urovirulence. Here, we sought to determine the uropathogenic properties of the Copenhagen outbreak strain and whether these properties are conferred by the EAEC-specific virulence factors. We demonstrated that through expression of aggregative adherence fimbriae, the principal adhesins of EAEC, the outbreak strain exhibited pronouncedly increased adherence to human bladder epithelial cells compared to prototype uropathogenic strains. Moreover, the strain was able to produce distinct biofilms on abiotic surfaces, including urethral catheters. These findings suggest that EAEC-specific virulence factors increase uropathogenicity and may have played a significant role in the ability of the strain to cause a community-acquired outbreak of UTI. Thus, inclusion of EAEC-specific virulence factors is warranted in future detection and characterization of uropathogenic E. coli. PMID:23357383

Boll, Erik J.; Struve, Carsten; Boisen, Nadia; Olesen, Bente; Stahlhut, Steen G.

2013-01-01

107

Genetic recombination. [Escherichia coli  

SciTech Connect

The molecular pathways of gene recombination are explored and compared in studies of the model organisms, Escherichia coli and phase lambda. In the discussion of data from these studies it seems that recombination varies with the genetic idiosyncrasies of the organism and may also vary within a single organism.

Stahl, F.W.

1987-02-01

108

Screening of SdiA inhibitors from Melia dubia seeds extracts towards the hold back of uropathogenic E.coli quorum sensing-regulated factors.  

PubMed

Plants have always been a supreme source of drugs and India is endowed with a wide variety of them with high medicinal values. The Quorum Sensing (QS) quenching efficiency of various solvent extracts of Melia dubia seeds was investigated against uropathogenic Escherichia coli (UPEC) to screen the competitive inhibitor of SdiA, a transcriptional activator of quorum sensing in E. coli. In this study, potentiality of five different extracts of Melia dubia seeds for quorum sensing inhibitory activity was investigated against uropathogenic Escherichia coli (UPEC). Assays such as cell density, swarming motility, protein, protease, hemolysis, hemagglutination, hydrophobicity and biofilm inhibition were performed. Biofilm, hemolysis and swarming motility were found to be inhibited by 92.1%, 20.9 % and 48.52% respectively, when the medium was supplemented with 30 mg/ml of the ethanolic extract. GC-MS spectrum of the ethanolic extract showed an array of 27 structurally unlinked compounds with natural ligand C8HSL. The docking against QS transcriptional regulator SdiA was predicted by in silico studies and the ligand C6 showed significant activity with -10.8 GScore. In vitro and in silico docking analysis showed fairly a good correlation, suggesting that the ethanolic extract showed potency to attenuate quorum sensing of uropathogenic E. coli. Further studies by in vitro and in vivo strategies are necessary to foresee the quorum quenching effect of the ligands. PMID:23210902

Ravichandiran, Vinothkannan; Shanmugam, Karthi; Solomon, Adline Princy

2013-09-01

109

Recurrent Escherichia coli bacteremia.  

PubMed

Escherichia coli is the most common gram-negative organism associated with bacteremia. While recurrent E. coli urinary tract infections are well-described, recurrent E. coli bacteremia appears to be uncommon, with no episodes noted in multiple series of patients with gram-negative bacteremias. We report on 5 patients with recurrent bloodstream infections identified from a series of 163 patients with E. coli bacteremia. For each patient, the isolates from each episode were analyzed by pulsed-field gel electrophoresis (PFGE) and ribotyping and for the presence of E. coli virulence factors. For each of four patients, the index and recurrent episodes of bacteremia represented the same strain as defined by PFGE, and the strains were found to carry one or more virulence factors. The remaining patient, with two episodes of bloodstream infection separated by a 4-year interval, was infected with two isolates that did not carry any virulence factors and that were clonally related by ribotype analysis but differed by PFGE. All five patients had either a local host defense defect (three patients) or impaired systemic defenses (one patient) or both (one patient). Thus, recurrent E. coli bacteremia is likely to represent a multifactorial process that occurs in patients with impaired host defenses who are infected with virulent isolates. PMID:7910828

Maslow, J N; Mulligan, M E; Arbeit, R D

1994-03-01

110

Aging of Escherichia coli  

PubMed Central

Clifton, C. E. (Stanford University, Stanford, Calif.). Aging of Escherichia coli. J. Bacteriol. 92:905–912. 1966.—The rates of endogenous and exogenous (glucose) respiration decreased much more rapidly than did the viable count during the first 24 hr of aging of washed, C14-labeled cells of Escherichia coli K-12 suspended in a basal salt medium devoid of ammonium salts. The rates of decrease of respiration and of death approached each other as the age of the cells increased, but death was not the only factor involved in decreased respiratory activity of the suspensions. The greatest decrease in cellular contents with aging was noted in the ribonucleic acid fraction, of which the ribose appeared to be oxidized, while uracil accumulated in the suspension medium. The viable count and respiratory activities remained higher in glucose-fed than in nonfed suspensions. Proline-labeled cells fed glucose tended to lose more of their proline and to convert more proline into C14O2 than in unfed controls. On the other hand, uracil-labeled cells fed glucose retained more of the uracil than did nonfed cells, but glucose elicited some oxidation of uracil. An exogenous energy source such as glucose aided in the maintenance of a population, but it was not the only factor needed for such maintenance. PMID:5332874

Clifton, C. E.

1966-01-01

111

PATHOGENIC ESCHERICHIA COLI IN FOODS  

Technology Transfer Automated Retrieval System (TEKTRAN)

Pathogenic Escherichia coli are defined as those E. coli strains that are capable of causing diarrhoeal disease in humans. Subdivision of the pathogenic forms is made on the basis of the mechanism underlying the illness. Presently, four types of pathogenic E. coli have been implicated in foodborne...

112

Contact-dependent inhibition of growth in Escherichia coli.  

PubMed

Bacteria have developed mechanisms to communicate and compete with each other for limited environmental resources. We found that certain Escherichia coli, including uropathogenic strains, contained a bacterial growth-inhibition system that uses direct cell-to-cell contact. Inhibition was conditional, dependent upon the growth state of the inhibitory cell and the pili expression state of the target cell. Both a large cell-surface protein designated Contact-dependent inhibitor A (CdiA) and two-partner secretion family member CdiB were required for growth inhibition. The CdiAB system may function to regulate the growth of specific cells within a differentiated bacterial population. PMID:16109881

Aoki, Stephanie K; Pamma, Rupinderjit; Hernday, Aaron D; Bickham, Jessica E; Braaten, Bruce A; Low, David A

2005-08-19

113

Biotyping of Escherichia coli.  

PubMed

We examined the results of tests with 22 substrates for their ability to discriminate a series of 917 strains of Escherichia coli collected from different sources. The tests with three of the substrates were discarded because of difficulties in performance or interpretation, and another nine substrates because they provided little discrimination. The tests used to obtain biotype profiles for strains were those for the fermentation of dulcitol, D-raffinose or sucrose or both, L-rhamnose and L-sorbose, the decarboxylation of L-lysine and L-ornithine, the hydrolysis of aesculin, motility, and prototrophy. Observations on several series of cultures from different sources showed that biotype characters were stable in vivo and after storage on non-selective medium. The biotype profiles obtained were as reliable as partial O serotyping for the routine subtyping of strains of E. coli isolated from the urine of patients with long-term urinary-tract infections and those from other sources in different patients. Biotyping and O serotyping used in conjuction offered a very fine degree of strain discrimination. PMID:390154

Crichton, P B; Old, D C

1979-11-01

114

Antibiotic Resistance in Uropathogenic E. Coli Strains Isolated from Non-Hospitalized Patients in Pakistan  

PubMed Central

Purpose: To study multidrug-resistance in Uropathogenic E. Coli (UPEC) isolated from non-hospitalized patients. Materials and Methods: Altogether, 250 bacterial samples were collected from non-hospitalized patients. Their identifications were done on basis of Gram-staining, colony morphology, biochemical testing and PCR. Susceptibility testing was performed by using standard protocols which were recommended by CLSI. Statistical analysis: For comparisons, statistical analysis was performed by using software, Graphpad Prism 5.0 Results: In total, 32% (n = 80) of the isolates were identified as E. Coli strains and their susceptibility patterns for different antibiotics were determined. The data indicated least resistance against tazocin [(TZP) -1.25%], amikacin [(AK) -1.8%], tigecycline [(TGC)- 2.5%] and nitrofurantoin [(F) -3.75%]. For both minocycline (MH) and sulzone (SUL), resistance rate was 5%, for gentamicin (CN), it was 16.25%, while higher resistances were observed against cephalothine [(KF)- 70%], cefotaxime [(CTX) -58.5%], ceftazidime [(CAZ)- 57.5%], cefepime [(FEP) -55%], cefuroxime and cefixime [(CXM) (CFM)- 53.75 %]. Resistance against ciprofloxacin (CIP) was 57.5%, for norfloxacine (NOR), it was 52.5% and incase of sparfloxacin (SPX), it remained 55%. High percentage of the isolates were resistant to cotrimoxazole [(SXT) -86%] and Amoxicillin [AMX-CLA (AMC)- 76%]. No resistance against meropenem (MEM) was observed. Conclusion: Highest level of drug-resistance was observed against trimethoprim-sulfamethoxazole (TMP-SMZ) among clinical isolates of uropathogenic E. Coli collected from non-hospitalized patients. PMID:25386430

Ali, Ihsan; Kumar, Neeraj; Ahmed, Safia

2014-01-01

115

Detection of urovirulence factors in Escherichia coli by multiplex polymerase chain reaction.  

PubMed

Primers to amplify the genes encoding the virulence factors of uropathogenic Escherichia coli, such as pilus associated with pyelonephritis (pap), haemolysin (hly), aerobactin (aer) and cytotoxic necrotizing factor 1 (cnf1) genes, were designed. The above primers along with previously reported primers for S fimbriae (sfa) and afimbrial adhesin I (afaI) genes were combined to develop a multiplex polymerase chain reaction (PCR) for detection of the respective virulence factors and for the identification of uropathogenic E. coli. The multiplex PCR to detect pap, sfa, afaI, hly, aer and cnf1 genes was highly specific and the sensitivity was found to be about 5 x 10(3) colony forming units of E. coli per ml. A total of 194 E. coli strains isolated from patients with simple acute cystitis were examined by the multiplex PCR and the results were in complete agreement with that obtained by DNA colony hybridization test. The multiplex PCR developed was, therefore, concluded to be a useful, sensitive and rapid assay system to identify uropathogenic E. coli. PMID:8589667

Yamamoto, S; Terai, A; Yuri, K; Kurazono, H; Takeda, Y; Yoshida, O

1995-10-01

116

Review article Enterotoxigenic Escherichia coli (ETEC)  

E-print Network

Review article Enterotoxigenic Escherichia coli (ETEC) in farm animals Béla Nagy* Péter Zs. Fekete to enterotoxigenic Escherichia coli (ETEC) typically appear as severe watery diarrhoea during the first few days a population of E. coli with a diverse genetic background. @ Inra/Elsevier, Paris. enterotoxin / fimbria

Paris-Sud XI, Université de

117

Host-Pathogen Checkpoints and Population Bottlenecks in Persistent and Intracellular Uropathogenic E. coli Bladder Infection  

PubMed Central

Bladder infections affect millions of people yearly, and recurrent symptomatic infections (cystitis) are very common. The rapid increase in infections caused by multi-drug resistant uropathogens threatens to make recurrent cystitis an increasingly troubling public health concern. Uropathogenic E. coli (UPEC) cause the vast majority of bladder infections. Upon entry into the lower urinary tract, UPEC face obstacles to colonization that constitute population bottlenecks, reducing diversity and selecting for fit clones. A critical mucosal barrier to bladder infection is the epithelium (urothelium). UPEC bypass this barrier when they invade urothelial cells and form intracellular bacterial communities (IBCs), a process which requires type 1 pili. IBCs are transient in nature, occurring primarily during acute infection. Chronic bladder infection is common and can be either latent, in the form of the Quiescent Intracellular Reservoir (QIR), or active, in the form of asymptomatic bacteriuria (ASB/ABU) or chronic cystitis. In mice, the fate of bladder infection: QIR, ASB, or chronic cystitis, is determined within the first 24 hours of infection and constitutes a putative host-pathogen mucosal checkpoint that contributes to susceptibility to recurrent cystitis. Knowledge of these checkpoints and bottlenecks is critical for our understanding of bladder infection and efforts to devise novel therapeutic strategies. PMID:22404313

Hannan, Thomas J.; Totsika, Makrina; Mansfield, Kylie J.; Moore, Kate H.; Schembri, Mark A.; Hultgren, Scott J.

2013-01-01

118

Characterization of urinary tract infection-associated Shiga toxin-producing Escherichia coli.  

PubMed

Enterohemorrhagic Escherichia coli (EHEC), a subgroup of Shiga toxin (Stx)-producing E. coli (STEC), is a leading cause of diarrhea and hemolytic-uremic syndrome (HUS) in humans. However, urinary tract infections (UTIs) caused by this microorganism but not associated with diarrhea have occasionally been reported. We geno- and phenotypically characterized three EHEC isolates obtained from the urine of hospitalized patients suffering from UTIs. These isolates carried typical EHEC virulence markers and belonged to HUS-associated E. coli (HUSEC) clones, but they lacked virulence markers typical of uropathogenic E. coli. One isolate exhibited a localized adherence (LA)-like pattern on T24 urinary bladder epithelial cells. Since the glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer) are well-known receptors for Stx but also for P fimbriae, a major virulence factor of extraintestinal pathogenic E. coli (ExPEC), the expression of Gb3Cer and Gb4Cer by T24 cells and in murine urinary bladder tissue was examined by thin-layer chromatography and mass spectrometry. We provide data indicating that Stxs released by the EHEC isolates bind to Gb3Cer and Gb4Cer isolated from T24 cells, which were susceptible to Stx. All three EHEC isolates expressed stx genes upon growth in urine. Two strains were able to cause UTI in a murine infection model and could not be outcompeted in urine in vitro by typical uropathogenic E. coli isolates. Our results indicate that despite the lack of ExPEC virulence markers, EHEC variants may exhibit in certain suitable hosts, e.g., in hospital patients, a uropathogenic potential. The contribution of EHEC virulence factors to uropathogenesis remains to be further investigated. PMID:25156739

Toval, Francisco; Schiller, Roswitha; Meisen, Iris; Putze, Johannes; Kouzel, Ivan U; Zhang, Wenlan; Karch, Helge; Bielaszewska, Martina; Mormann, Michael; Müthing, Johannes; Dobrindt, Ulrich

2014-11-01

119

Effect of 2,4-Dichlorophenoxyacetic acid herbicide Escherichia coli growth, chemical, composition, and cellular envelope  

USGS Publications Warehouse

2,4-Dichlorophenoxyacetic acid (2,4-D) is a herbicide widely used in the world and mainly excreted by the renal route in exposed humans and animals. Herbicides can affect other nontarget organisms, such as Escherichia coli. We observed that a single exposure to 1 mM 2,4-D diminished growth and total protein content in all E. coli strains tested in vitro. In addition, successive exposures to 0.01 mM 2,4-D had a toxic effect decreasing growth up to early stationary phase. Uropathogenic E. coli adhere to epithelial cells mediated by fimbriae, adhesins, and hydrophobic properties. 2,4-D exposure of uropathogenic E. coli demonstrated altered hydrophobicity and fimbriation. Hydrophobicity index values obtained by partition in p-xylene/water were 300-420% higher in exposed cells than in control ones. Furthermore, values of hemagglutination titer, protein contents in fimbrial crude extract, and electron microscopy demonstrated a significant diminution of fimbriation in treated cells. Other envelope alterations could be detected, such as lipoperoxidation, evidenced by decreased polyunsaturated fatty acids and increased lipid degradation products (malonaldehyde), and motility diminution. These alterations decreased cell adherence to erythrocytes, indicating a diminished pathogenic capacity of the 2,4-D-exposed E. coli. ?? 2001 by John Wiley & Sons, Inc.

Carr, R.S.; Biedenbach, J.M.; Hooten, R.L.

2001-01-01

120

Cardiolipin synthase from Escherichia coli.  

PubMed

Escherichia coli cardiolipin synthase catalyzes reversible phosphatidyl group transfer from one phosphatidylglycerol molecule to another to form cardiolipin (CL) and glycerol. The enzyme is specified by the cls gene, located at min 28.02 of the E. coli genetic map. Cells with mutations in cls have longer doubling times, tend to lose viability in the stationary phase, are more resistant to 3,4-dihydroxybutyl-1-phosphonate, and have an altered sensitivity to novobiocin. Although cls null mutants appear to lack CL synthase activity, they are still able to form trace quantities of CL. The enzyme appears to be regulated at both the genetic and enzymatic levels. CL synthase's molecular mass is 45-46 kDa, or about 8 kDa less than the polypeptide predicted by the gene sequence, suggesting that posttranslational processing occurs. CL synthase can use various polyols such as mannitol and arabitol to convert CL to the corresponding phosphatidylglycerol analog. When the amino acid sequences of four bacterial CL synthases are compared, three highly conserved regions are apparent. One of these regions contains a conserved pentapeptide sequence, RN(Q)HRK, and another has a conserved HXK sequence. These two sequences may be part of the active site. E. coli CL synthase has been studied by using a mixed micelle assay. The enzyme is inhibited by CL, the product of the reaction, and by phosphatidate. Phosphatidylethanolamine partially offsets inhibition caused by CL but not by phosphatidate. CDP-diacylglycerol does not appear to affect the activity of the purified enzyme but does stimulate the activity associated with crude membrane preparations. PMID:9370333

Tropp, B E

1997-09-01

121

Recombinant collagen production optimization in Escherichia coli  

E-print Network

An Escherichia coli-based collagen-production process was used to investigate several process optimization objectives for use at the industrial scale. The effect of cooling on fermentation growth kinetics was studied, with ...

Whittemore, Brett A

2005-01-01

122

High level indole signalling in Escherichia coli  

E-print Network

Chapter 1 Introduction 16 1.1 Bacterial Communication and Signalling Bacteria are able to produce and use a wide variety of chemical signals to communicate. They are able to utilise these messages to better respond to changing environments... indole signalling in Escherichia coli Abstract Indole is a small signalling molecule, produced by many species of bacteria, including Escherichia coli. It is made by the enzyme tryptophanase, which converts tryptophan into indole, pyruvate and ammonia...

Gaimster, Hannah Dorne

2014-06-10

123

Characterization of intestinal cnf1+ Escherichia coli from weaned pigs.  

PubMed

Escherichia coli isolated from 204 cases of porcine postweaning diarrhoea were tested by PCR for the genes of cytotoxic necrotic factors (CNF) and of cytolethal dystending toxin (CDT). Selected strains were also examined by PCR for the presence of papC-, sfa-, f17-, f18-, and afa-specific sequences encoding P, S, F17, F18 fimbriae and afimbrial adhesins. A 5.9% (12/204) of the strains had cnf1 gene, and two of them had cdt gene as well. Further six cdt+ strains were detected which were cnf-negative. Most of the cnf1+ strains belonged to serogroups O2, O6, O8, O54 characteristic of necrotoxic E. coli (NTEC) of humans. All the cnf1+ strains possessed the genes for P or S fimbriae or both, but were negative for F4, F17, or F18 or afimbrial adhesins. Results suggest that these enteric isolates may have entero- and/or uropathogenic significance in weaned pigs, and may have zoonotic potential for humans. PMID:11100828

Tóth, I; Oswald, E; Mainil, J G; Awad-Masalmeh, M; Nagy, B

2000-10-01

124

Strategies for Protein Overproduction in Escherichia coli.  

ERIC Educational Resources Information Center

Examines heterologous expression in Escherichia coli and the role of regulatory sequences which control gene expression at transcription resulting in abundant production of messenger RNA and regulatory sequences in mRNA which promote efficient translation. Also examines the role of E. coli cells in stabilizing mRNA and protein that is…

Mott, John E.

1984-01-01

125

Frequency of Escherichia coli strains producing the cytotoxic necrotizing factor (CNF1) in nosocomial urinary tract infections.  

PubMed

The presence of cytotoxic necrotizing factor 1 (CNF1), together with various associated virulence factors (alpha-haemolysin, P-, S- and A-fimbriae), was screened in 175 uropathogenic Escherichia coli strains isolated from hospitalized adult patients. The cnf1 gene was detected in 30% of the selected strains independently of the severity of the clinical urinary infection. A significant association between CNF1, haemolytic activity and the products of the pap/sfa genes was found. However, CNF1 appeared not to play a major role in nosocomial E. coli urinary tract infections. PMID:10747253

Landraud, L; Gauthier, M; Fosse, T; Boquet, P

2000-03-01

126

Succinate production in Escherichia coli  

PubMed Central

Succinate has been recognized as an important platform chemical that can be produced from biomass. While a number of organisms are capable of succinate production naturally, this review focuses on the engineering of Escherichia coli for production of the four-carbon dicarboxylic acid. Important features of a succinate production system are to achieve optimal balance of reducing equivalents generated by consumption of the feedstock, while maximizing the amount of carbon that is channeled to the product. Aerobic and anaerobic production strains have been developed and applied to production from glucose as well as other abundant carbon sources. Metabolic engineering methods and strain evolution have been used and supplemented by the recent application of systems biology and in silico modeling tools to construct optimal production strains. The metabolic capacity of the production strain, as well as the requirement for efficient recovery of succinate and the reliability of the performance under scale-up are important in the overall process. The costs of the overall biorefinery compatible process will determine the economical commercialization of succinate and its impact in larger chemical markets. PMID:21932253

Thakker, Chandresh; Martínez, Irene; San, Ka-Yiu; Bennett, George N.

2012-01-01

127

Asparagine Utilization in Escherichia coli  

PubMed Central

Asparagine-requiring auxotrophs of Escherichia coli K-12 that have an active cytoplasmic asparaginase do not conserve asparagine supplements for use in protein synthesis. Asparagine molecules entering the cell in excess of the pool required for use of this amino acid in protein synthesis are rapidly degraded rather than accumulated. Supplements are conserved when asparagine degradation is inhibited by the asparagine analogue 5-diazo-4-oxo-l-norvaline (DONV) or mutation to cytoplasmic asparaginase deficiency. A strain deficient in cytoplasmic asparaginase required approximately 260 ?mol of asparagine for the synthesis of 1 g of cellular protein. The cytoplasmic asparaginase (asparaginase I) is required for growth of cells when asparagine is the nitrogen source. This enzyme has an apparent Km for l-asparagine of 3.5 mM, and asparaginase activity is competitively inhibited by DONV with an apparent Ki of 2 mM. The analogue provides a time-dependent, irreversible inhibition of cytoplasmic asparaginase activity in the absence of asparagine. PMID:4595199

Willis, R. C.; Woolfolk, C. A.

1974-01-01

128

A draft genome of Escherichia coli sequence type 127 strain 2009-46  

PubMed Central

Background Escherichia coli are a frequent cause of urinary tract infections (UTI) and are thought to have a foodborne origin. E. coli with sequence type 127 (ST127) are emerging pathogens increasingly implicated as a cause of urinary tract infections (UTI) globally. A ST127 isolate (2009-46) resistant to ampicillin and trimethoprim was recovered from the urine of a 56 year old patient with a UTI from a hospital in Sydney, Australia and was characterised here. Results We sequenced the genome of Escherichia coli 2009-46 using the Illumina Nextera XT and MiSeq technologies. Assembly of the sequence data reconstructed a 5.14 Mbp genome in 89 scaffolds with an N50 of 161 kbp. The genome has extensive similarity to other sequenced uropathogenic E. coli genomes, but also has several genes that are potentially related to virulence and pathogenicity that are not present in the reference E. coli strain. Conclusion E. coli 2009-46 is a multiple antibiotic resistant, phylogroup B2 isolate recovered from a patient with a UTI. This is the first description of a drug resistant E. coli ST127 in Australia. PMID:25197321

2014-01-01

129

Galleria mellonella Infection Model Demonstrates High Lethality of ST69 and ST127 Uropathogenic E. coli  

PubMed Central

Galleria mellonella larvae are an alternative in vivo model for investigating bacterial pathogenicity. Here, we examined the pathogenicity of 71 isolates from five leading uropathogenic E. coli (UPEC) lineages using G. mellonella larvae. Larvae were challenged with a range of inoculum doses to determine the 50% lethal dose (LD50) and for analysis of survival outcome using Kaplan-Meier plots. Virulence was correlated with carriage of a panel of 29 virulence factors (VF). Larvae inoculated with ST69 and ST127 isolates (104 colony-forming units/larvae) showed significantly higher mortality rates than those infected with ST73, ST95 and ST131 isolates, killing 50% of the larvae within 24 hours. Interestingly, ST131 isolates were the least virulent. We observed that ST127 isolates are significantly associated with a higher VF-score than isolates of all other STs tested (P?0.0001), including ST69 (P<0.02), but one ST127 isolate (strain EC18) was avirulent. Comparative genomic analyses with virulent ST127 strains revealed an IS1 mediated deletion in the O-antigen cluster in strain EC18, which is likely to explain the lack of virulence in the larvae infection model. Virulence in the larvae was not correlated with serotype or phylogenetic group. This study illustrates that G. mellonella are an excellent tool for investigation of the virulence of UPEC strains. The findings also support our suggestion that the incidence of ST127 strains should be monitored, as these isolates have not yet been widely reported, but they clearly have a pathogenic potential greater than that of more widely recognised clones, including ST73, ST95 or ST131. PMID:25061819

Alghoribi, Majed F.; Gibreel, Tarek M.; Dodgson, Andrew R.; Beatson, Scott A.; Upton, Mathew

2014-01-01

130

Infection strategies of enteric pathogenic Escherichia coli  

PubMed Central

Enteric Escherichia coli (E. coli) are both natural flora of humans and important pathogens causing significant morbidity and mortality worldwide. Traditionally enteric E. coli have been divided into 6 pathotypes, with further pathotypes often proposed. In this review we suggest expansion of the enteric E. coli into 8 pathotypes to include the emerging pathotypes of adherent invasive E. coli (AIEC) and Shiga-toxin producing enteroaggregative E. coli (STEAEC). The molecular mechanisms that allow enteric E. coli to colonize and cause disease in the human host are examined and for two of the pathotypes that express a type 3 secretion system (T3SS) we discuss the complex interplay between translocated effectors and manipulation of host cell signaling pathways that occurs during infection. PMID:22555463

Clements, Abigail; Young, Joanna C.; Constantinou, Nicholas; Frankel, Gad

2012-01-01

131

Virulence profile of different phylogenetic groups of locally isolated community acquired uropathogenic E. coli from Faisalabad region of Pakistan  

PubMed Central

Background Uropathogenic E.coli (UPEC) are among major pathogens causing urinary tract infections. Virulence factors are mainly responsible for the severity of these emerging infections. This study was planned to investigate the distribution of virulence genes and cytotoxic effects of UPEC isolates with reference to phylogenetic groups (B2, B1, D and A) to understand the presence and impact of virulence factors in the severity of infection in Faisalabad region of Pakistan. Methods In this study phylogenetic analysis, virulence gene identification and cytotoxicity of 59 uropathogenic E.coli isolates obtained from non-hospitalized patients was studied. Results Among 59 isolates, phylogenetic group B2 (50%) was most dominant followed by groups A, B1 (19% each) and D (12%). Isolates present in group D showed highest presence of virulence genes. The prevalence hlyA (37%) was highest followed by sfaDE (27%), papC (24%), cnf1 (20%), eaeA (19%) and afaBC3 (14%). Highly hemolytic and highly verotoxic isolates mainly belonged to group D and B2. We also found two isolates with simultaneous presence of three fimbrial adhesin genes present on pap, afa, and sfa operons. This has not been reported before and underlines the dynamic nature of these UPEC isolates. Conclusions It was concluded that in local UPEC isolates from non-hospitalized patients, group B2 was more prevalent. However, group D isolates were most versatile as all were equipped with virulence genes and showed highest level of cytotoxicity. PMID:22867028

2012-01-01

132

Virulence factors in Escherichia coli urinary tract infection.  

PubMed Central

Uropathogenic strains of Escherichia coli are characterized by the expression of distinctive bacterial properties, products, or structures referred to as virulence factors because they help the organism overcome host defenses and colonize or invade the urinary tract. Virulence factors of recognized importance in the pathogenesis of urinary tract infection (UTI) include adhesins (P fimbriae, certain other mannose-resistant adhesins, and type 1 fimbriae), the aerobactin system, hemolysin, K capsule, and resistance to serum killing. This review summarizes the virtual explosion of information regarding the epidemiology, biochemistry, mechanisms of action, and genetic basis of these urovirulence factors that has occurred in the past decade and identifies areas in need of further study. Virulence factor expression is more common among certain genetically related groups of E. coli which constitute virulent clones within the larger E. coli population. In general, the more virulence factors a strain expresses, the more severe an infection it is able to cause. Certain virulence factors specifically favor the development of pyelonephritis, others favor cystitis, and others favor asymptomatic bacteriuria. The currently defined virulence factors clearly contribute to the virulence of wild-type strains but are usually insufficient in themselves to transform an avirulent organism into a pathogen, demonstrating that other as-yet-undefined virulence properties await discovery. Virulence factor testing is a useful epidemiological and research tool but as yet has no defined clinical role. Immunological and biochemical anti-virulence factor interventions are effective in animal models of UTI and hold promise for the prevention of UTI in humans. Images PMID:1672263

Johnson, J R

1991-01-01

133

Iron induces bimodal population development by Escherichia coli.  

PubMed

Bacterial biofilm formation is a complex developmental process involving cellular differentiation and the formation of intricate 3D structures. Here we demonstrate that exposure to ferric chloride triggers rugose biofilm formation by the uropathogenic Escherichia coli strain UTI89 and by enteric bacteria Citrobacter koseri and Salmonella enterica serovar typhimurium. Two unique and separable cellular populations emerge in iron-triggered, rugose biofilms. Bacteria at the air-biofilm interface express high levels of the biofilm regulator csgD, the cellulose activator adrA, and the curli subunit operon csgBAC. Bacteria in the interior of rugose biofilms express low levels of csgD and undetectable levels of matrix components curli and cellulose. Iron activation of rugose biofilms is linked to oxidative stress. Superoxide generation, either through addition of phenazine methosulfate or by deletion of sodA and sodB, stimulates rugose biofilm formation in the absence of high iron. Additionally, overexpression of Mn-superoxide dismutase, which can mitigate iron-derived reactive oxygen stress, decreases biofilm formation in a WT strain upon iron exposure. Not only does reactive oxygen stress promote rugose biofilm formation, but bacteria in the rugose biofilms display increased resistance to H(2)O(2) toxicity. Altogether, we demonstrate that iron and superoxide stress trigger rugose biofilm formation in UTI89. Rugose biofilm development involves the elaboration of two distinct bacterial populations and increased resistance to oxidative stress. PMID:23359678

DePas, William H; Hufnagel, David A; Lee, John S; Blanco, Luz P; Bernstein, Hans C; Fisher, Steve T; James, Garth A; Stewart, Philip S; Chapman, Matthew R

2013-02-12

134

Inhibition of biofilm development of uropathogens by curcumin - an anti-quorum sensing agent from Curcuma longa.  

PubMed

Urinary tract infection is caused primarily by the quorum sensing (QS)-dependent biofilm forming ability of uropathogens. In the present investigation, an anti-quorum sensing (anti-QS) agent curcumin from Curcuma longa (turmeric) was shown to inhibit the biofilm formation of uropathogens, such as Escherichia coli, Pseudomonas aeruginosa PAO1, Proteus mirabilis and Serratia marcescens, possibly by interfering with their QS systems. The antibiofilm potential of curcumin on uropathogens as well as its efficacy in disturbing the mature biofilms was examined under light microscope and confocal laser scanning microscope. The treatment with curcumin was also found to attenuate the QS-dependent factors, such as exopolysaccharide production, alginate production, swimming and swarming motility of uropathogens. Furthermore, it was documented that curcumin enhanced the susceptibility of a marker strain and uropathogens to conventional antibiotics. PMID:24262582

Packiavathy, Issac Abraham Sybiya Vasantha; Priya, Selvam; Pandian, Shunmugiah Karutha; Ravi, Arumugam Veera

2014-04-01

135

Native valve Escherichia coli endocarditis following urosepsis  

PubMed Central

Gram-negative organisms are a rare cause of infective endocarditis. Escherichia coli, the most common cause of urinary tract infection and gram-negative septicemia involves endocardium rarely. In this case report, we describe infection of native mitral valve by E. coli following septicemia of urinary tract origin in a diabetic male; subsequently, he required prosthetic tissue valve replacement indicated by persistent sepsis and congestive cardiac failure. PMID:23814428

Rangarajan, D.; Ramakrishnan, S.; Patro, K. C.; Devaraj, S.; Krishnamurthy, V.; Kothari, Y.; Satyaki, N.

2013-01-01

136

Chaperone-Usher Fimbriae of Escherichia coli  

PubMed Central

Chaperone-usher (CU) fimbriae are adhesive surface organelles common to many Gram-negative bacteria. Escherichia coli genomes contain a large variety of characterised and putative CU fimbrial operons, however, the classification and annotation of individual loci remains problematic. Here we describe a classification model based on usher phylogeny and genomic locus position to categorise the CU fimbrial types of E. coli. Using the BLASTp algorithm, an iterative usher protein search was performed to identify CU fimbrial operons from 35 E. coli (and one Escherichia fergusonnii) genomes representing different pathogenic and phylogenic lineages, as well as 132 Escherichia spp. plasmids. A total of 458 CU fimbrial operons were identified, which represent 38 distinct fimbrial types based on genomic locus position and usher phylogeny. The majority of fimbrial operon types occupied a specific locus position on the E. coli chromosome; exceptions were associated with mobile genetic elements. A group of core-associated E. coli CU fimbriae were defined and include the Type 1, Yad, Yeh, Yfc, Mat, F9 and Ybg fimbriae. These genes were present as intact or disrupted operons at the same genetic locus in almost all genomes examined. Evaluation of the distribution and prevalence of CU fimbrial types among different pathogenic and phylogenic groups provides an overview of group specific fimbrial profiles and insight into the ancestry and evolution of CU fimbriae in E. coli. PMID:23382825

Wurpel, Daniël J.; Beatson, Scott A.; Totsika, Makrina; Petty, Nicola K.; Schembri, Mark A.

2013-01-01

137

Beta-alanine synthesis in Escherichia coli.  

PubMed Central

The enzyme, aspartate 1-decarboxylase (L-aspartate 1-carboxy-lyase; EC 4.1.1.15), that catalyzes the reaction aspartate leads to beta-alanine + CO2 was found in extracts of Escherichia coli. panD mutants of E. coli are defective in beta-alanine biosynthesis and lack aspartate 1-decarboxylase. Therefore, the enzyme functions in the biosynthesis of the beta-alanine moiety of pantothenate. The genetic lesion in these mutants is closely linked to the other pantothenate (pan) loci of E. coli K-12. Images PMID:6767707

Cronan, J E

1980-01-01

138

Difference in the regulation of IL-8 expression induced by uropathogenic E. coli between two kinds of urinary tract epithelial cells  

PubMed Central

Bacterial adherence to epithelial cells is a key virulence trait of pathogenic bacteria. The type 1 fimbriae and the P-fimbriae of uropathogenic Escherichia coli (UPEC) have both been described to be important for the establishment of urinary tract infections (UTI). To explore the interactions between the host and bacterium responsible for the different environments of UPEC invasion, we examined the effect of pH and osmolarity on UPEC strain J96 fimbrial expression, and subsequent J96-induced interleukin-8 (IL-8) expression in different uroepithelial cells. The J96 strain grown in high pH with low osmolarity condition was favorable for the expression of type 1 fimbriae; whereas J96 grown in low pH with high osmolarity condition was beneficial for P fimbriae expression. Type 1 fimbriated J96 specifically invaded bladder 5637 epithelial cells and induced IL-8 expression. On the contrary, P fimbriated J96 invaded renal 786-O epithelial cells and induced IL-8 expression effectively. Type 1 fimbriated J96-induced IL-8 induction involved the p38, as well as ERK, JNK pathways, which leads to AP-1-mediated gene expression. P fimbriated J96-induced augmentation of IL-8 expression mainly involved p38-mediated AP-1 and NF-?B transcriptional activation. These results indicate that different expression of fimbriae in J96 trigger differential IL-8 gene regulation pathways in different uroepithelial cells. PMID:19799797

2009-01-01

139

Pathogenic Escherichia coli Found in Sewage Treatment Plants and Environmental Waters  

PubMed Central

We previously demonstrated that some Escherichia coli strains with uropathogenic properties survived treatment stages of sewage treatment plants (STPs), suggesting that they may be released into the environment. We investigated the presence of such strains in the surrounding environmental waters of four STPs from which these persistent strains were isolated. In all, 264 E. coli isolates were collected from 129 receiving water sites in a 20-km radius surrounding STPs. We also included 93 E. coli strains collected from 18 animal species for comparison. Isolates were typed using a high-resolution biochemical fingerprinting method (the PhPlate system), and grouped into common (C) types. One hundred forty-seven (56%) environmental isolates were identical to strains found in STPs' final effluents. Of these, 140 (95%) carried virulence genes (VGs) associated with intestinal pathogenic E. coli (IPEC) or uropathogenic E. coli (UPEC) and were found in a variety of sites within areas sampled. Of the remaining 117 environmental strains not identical to STP strains, 105 belonged to 18 C types and 102 of them carried VGs found among IPEC or UPEC strains. These strains belonged mainly to phylogenetic groups A (A0 and A1) and B1 and to a lesser extent B22, B23, D1, and D2. Eight of 18 environmental C types, comprising 50 isolates, were also identical to bird strains. The presence of a high percentage of environmental E. coli in waters near STPs carrying VGs associated with IPEC and UPEC suggests that they may have derived from STP effluents and other nonpoint sources. PMID:22660714

Anastasi, E. M.; Matthews, B.; Stratton, H. M.

2012-01-01

140

Heteropathogenic virulence and phylogeny reveal phased pathogenic metamorphosis in Escherichia coli O2:H6.  

PubMed

Extraintestinal pathogenic and intestinal pathogenic (diarrheagenic) Escherichia coli differ phylogenetically and by virulence profiles. Classic theory teaches simple linear descent in this species, where non-pathogens acquire virulence traits and emerge as pathogens. However, diarrheagenic Shiga toxin-producing E. coli (STEC) O2:H6 not only possess and express virulence factors associated with diarrheagenic and uropathogenic E. coli but also cause diarrhea and urinary tract infections. These organisms are phylogenetically positioned between members of an intestinal pathogenic group (STEC) and extraintestinal pathogenic E. coli. STEC O2:H6 is, therefore, a 'heteropathogen,' and the first such hybrid virulent E. coli identified. The phylogeny of these E. coli and the repertoire of virulence traits they possess compel consideration of an alternate view of pathogen emergence, whereby one pathogroup of E. coli undergoes phased metamorphosis into another. By understanding the evolutionary mechanisms of bacterial pathogens, rational strategies for counteracting their detrimental effects on humans can be developed. PMID:24413188

Bielaszewska, Martina; Schiller, Roswitha; Lammers, Lydia; Bauwens, Andreas; Fruth, Angelika; Middendorf, Barbara; Schmidt, M Alexander; Tarr, Phillip I; Dobrindt, Ulrich; Karch, Helge; Mellmann, Alexander

2014-03-01

141

Escherichia Coli--Key to Modern Genetics.  

ERIC Educational Resources Information Center

Mid-nineteenth century work by Mendel on plant hybrids and by Pasteur on fermentation gave birth by way of bacterial genetics to modern-day molecular biology. The bacterium Escherichia Coli has occupied a key position in genetic studies leading from early gene identification with DNA to current genetic engineering using recombinant DNA technology.…

Bregegere, Francois

1982-01-01

142

Maltoheptaose Promotes Nanoparticle Internalization by Escherichia coli  

PubMed Central

Nanoparticles conjugated with D-maltoheptaose (G7) showed a striking increase in the internalization by Escherichia coli. This applies to strains with and without the maltodextrin transport channel and particles ranging from a few to a hundred nanometers. PMID:23463337

Jayawardena, Surangi; Jayawardana, Kalana; Chen, Xuan

2013-01-01

143

Engineering ethanologenic Escherichia coli for levoglucosan utilization  

Microsoft Academic Search

Levoglucosan is a major product of biomass pyrolysis. While this pyrolyzed biomass, also known as bio-oil, contains sugars that are an attractive fermentation substrate, commonly-used biocatalysts, such as Escherichia coli, lack the ability to metabolize this anhydrosugar. It has previously been shown that recombinant expression of the levoglucosan kinase enzyme enables use of levoglucosan as carbon and energy source. Here,

Donovan S. Layton; Avanthi Ajjarapu; Dong Won Choi; Laura R. Jarboe

2011-01-01

144

Escherichia coli in Europe: An Overview  

PubMed Central

Escherichia coli remains one of the most frequent causes of several common bacterial infections in humans and animals. E. coli is the prominent cause of enteritis, urinary tract infection, septicaemia and other clinical infections, such as neonatal meningitis. E. coli is also prominently associated with diarrhoea in pet and farm animals. The therapeutic treatment of E. coli infections is threatened by the emergence of antimicrobial resistance. The prevalence of multidrug-resistant E. coli strains is increasing worldwide principally due to the spread of mobile genetic elements, such as plasmids. The rise of multidrug-resistant strains of E. coli also occurs in Europe. Therefore, the spread of resistance in E. coli is an increasing public health concern in European countries. This paper summarizes the current status of E. coli strains clinically relevant in European countries. Furthermore, therapeutic interventions and strategies to prevent and control infections are presented and discussed. The article also provides an overview of the current knowledge concerning promising alternative therapies against E. coli diseases. PMID:24287850

Allocati, Nerino; Masulli, Michele; Alexeyev, Mikhail F.; Di Ilio, Carmine

2013-01-01

145

Functional proteomics in Escherichia coli  

E-print Network

-state of E. coli corresponding to hundreds of unique gene products. The copurification of proteins when fractionated at varying pHs could suggest the components of higher order complexes. This non-denaturing proteomic approach should provide physiological...

Champion, Matthew Maurice

2006-04-12

146

Serogroups and virulence genotypes of Escherichia coli isolated from patients with sepsis.  

PubMed

Sixty strains of Escherichia coli, isolated by hemoculture, from septicemic Brazilian patients were evaluated to determine their serogroup and invasivity to Vero cells. All 60 patients died within 2 days of hospitalization. Furthermore, the molecular study of the following extraintestinal pathogenic E. coli-associated virulence factor (VF) genes was performed by PCR: i) adhesins: type 1 fimbria (fimH), S fimbria (sfaD/E), P fimbria (papC and papG alleles) and afimbrial adhesin (afaB/C); ii) capsule K1/K5 (kpsMTII); iii) siderophores: aerobactin (iucD), yersiniabactin (fyuA) and salmochelin (iroN); iv) toxins hemolysin (hlyA), necrotizing cytotoxic factor type 1 (cnf1) and secreted autotransporter toxin (sat); v) miscellaneous: brain microvascular endothelial cells invasion (ibeA), serum resistance (traT), colicin V (cvaC) and specific uropathogenic protein (usp). Our results showed that isolates are able to invade Vero cells (96.6%), differing from previous research on uropathogenic E. coli (UPEC). The O serogroups associated with UPEC were prevalent in 60% of strains vs 11.7% of other serogroups. The PCR results showed a conserved virulence subgroup profile and a prevalence above 75% for fimH, fyuA, kpsMTII and iucD, and between 35-65% for papC, papG, sat, iroN, usp and traT. The evasion from the immunological system of the host and also iron uptake are essential for the survival of extraintestinal pathogenic E. coli strains. Interestingly, among our isolates, a low prevalence of VF genes appeared. Therefore, the present study contributes to the identification of a bacterial profile for sepsis-associated E. coli. PMID:19030710

Ananias, M; Yano, T

2008-10-01

147

Pathogenesis of Afa/Dr Diffusely Adhering Escherichia coli  

PubMed Central

Over the last few years, dramatic increases in our knowledge about diffusely adhering Escherichia coli (DAEC) pathogenesis have taken place. The typical class of DAEC includes E. coli strains harboring AfaE-I, AfaE-II, AfaE-III, AfaE-V, Dr, Dr-II, F1845, and NFA-I adhesins (Afa/Dr DAEC); these strains (i) have an identical genetic organization and (ii) allow binding to human decay-accelerating factor (DAF) (Afa/DrDAF subclass) or carcinoembryonic antigen (CEA) (Afa/DrCEA subclass). The atypical class of DAEC includes two subclasses of strains; the atypical subclass 1 includes E. coli strains that express AfaE-VII, AfaE-VIII, AAF-I, AAF-II, and AAF-III adhesins, which (i) have an identical genetic organization and (ii) do not bind to human DAF, and the atypical subclass 2 includes E. coli strains that harbor Afa/Dr adhesins or others adhesins promoting diffuse adhesion, together with pathogenicity islands such as the LEE pathogenicity island (DA-EPEC). In this review, the focus is on Afa/Dr DAEC strains that have been found to be associated with urinary tract infections and with enteric infection. The review aims to provide a broad overview and update of the virulence aspects of these intriguing pathogens. Epidemiological studies, diagnostic techniques, characteristic molecular features of Afa/Dr operons, and the respective role of Afa/Dr adhesins and invasins in pathogenesis are described. Following the recognition of membrane-bound receptors, including type IV collagen, DAF, CEACAM1, CEA, and CEACAM6, by Afa/Dr adhesins, activation of signal transduction pathways leads to structural and functional injuries at brush border and junctional domains and to proinflammatory responses in polarized intestinal cells. In addition, uropathogenic Afa/Dr DAEC strains, following recognition of ?1 integrin as a receptor, enter epithelial cells by a zipper-like, raft- and microtubule-dependent mechanism. Finally, the presence of other, unknown virulence factors and the way that an Afa/Dr DAEC strain emerges from the human intestinal microbiota as a “silent pathogen” are discussed. PMID:15831825

Servin, Alain L.

2005-01-01

148

Prevalence and Antibiogram Profiling of Escherichia coli Pathotypes Isolated from the Kat River and the Fort Beaufort Abstraction Water  

PubMed Central

Escherichia coli is a widespread bacterium encompassing a variety of strains, ranging from highly pathogenic strains, causing worldwide outbreaks of severe diseases to avirulent, well characterized safe laboratory strains. This study evaluated the prevalence and antibiogram profiles of E. coli pathotypes isolated from the Kat River and Fort Beaufort abstraction water. A total of 171 out of 278 confirmed E. coli isolates were positive for at least one pathogenic determinant and these included enteropathogenic E. coli (6%), enterotoxigenic E. coli (47%), uropathogenic E. coli (2%), neonatal meningitis E. coli (5%), diffusely adherent E. coli (1%) and enterohaemorrhagic E. coli (1%). Interestingly, enteroinvasive and enteroaggregative E. coli were not detected. The phenotypic antibiogram profiles of the isolates revealed that all were resistant to penicillin G, while 98% and 38% of the pathotypes were resistant to ampicillin and trimethoprim-sulphamethoxazole, respectively. About 8% of the isolates were resistant to streptomycin. More than half of the isolates exhibited multiple antibiotic resistance with 44% being resistant to three antibiotics and 8% resistant to four antibiotics. We conclude that the Kat River is a reservoir of potentially virulent antibiotic resistant E. coli strains that can cause serious health risks to humans who drink raw water from this river, or in the case that consumption of treated drinking water coincides with failed drinking water processes. PMID:25119699

Nontongana, Nolonwabo; Sibanda, Timothy; Ngwenya, Elvis; Okoh, Anthony I.

2014-01-01

149

Asymptomatic bacteriuria Escherichia coli are live biotherapeutics for UTI.  

PubMed

Urinary tract infections (UTI) account for approximately 8 million clinic visits annually with symptoms that include acute pelvic pain, dysuria, and irritative voiding. Empiric UTI management with antimicrobials is complicated by increasing antimicrobial resistance among uropathogens, but live biotherapeutics products (LBPs), such as asymptomatic bacteriuria (ASB) strains of E. coli, offer the potential to circumvent antimicrobial resistance. Here we evaluated ASB E. coli as LBPs, relative to ciprofloxacin, for efficacy against infection and visceral pain in a murine UTI model. Visceral pain was quantified as tactile allodynia of the pelvic region in response to mechanical stimulation with von Frey filaments. Whereas ciprofloxacin promoted clearance of uropathogenic E. coli (UPEC), it did not reduce pelvic tactile allodynia, a measure of visceral pain. In contrast, ASB E. coli administered intravesically or intravaginally provided comparable reduction of allodynia similar to intravesical lidocaine. Moreover, ASB E. coli were similarly effective against UTI allodynia induced by Proteus mirabilis, Enterococccus faecalis and Klebsiella pneumoniae. Therefore, ASB E. coli have anti-infective activity comparable to the current standard of care yet also provide superior analgesia. These studies suggest that ASB E. coli represent novel LBPs for UTI symptoms. PMID:25405579

Rudick, Charles N; Taylor, Aisha K; Yaggie, Ryan E; Schaeffer, Anthony J; Klumpp, David J

2014-01-01

150

Asymptomatic Bacteriuria Escherichia coli Are Live Biotherapeutics for UTI  

PubMed Central

Urinary tract infections (UTI) account for approximately 8 million clinic visits annually with symptoms that include acute pelvic pain, dysuria, and irritative voiding. Empiric UTI management with antimicrobials is complicated by increasing antimicrobial resistance among uropathogens, but live biotherapeutics products (LBPs), such as asymptomatic bacteriuria (ASB) strains of E. coli, offer the potential to circumvent antimicrobial resistance. Here we evaluated ASB E. coli as LBPs, relative to ciprofloxacin, for efficacy against infection and visceral pain in a murine UTI model. Visceral pain was quantified as tactile allodynia of the pelvic region in response to mechanical stimulation with von Frey filaments. Whereas ciprofloxacin promoted clearance of uropathogenic E. coli (UPEC), it did not reduce pelvic tactile allodynia, a measure of visceral pain. In contrast, ASB E. coli administered intravesically or intravaginally provided comparable reduction of allodynia similar to intravesical lidocaine. Moreover, ASB E. coli were similarly effective against UTI allodynia induced by Proteus mirabilis, Enterococccus faecalis and Klebsiella pneumoniae. Therefore, ASB E. coli have anti-infective activity comparable to the current standard of care yet also provide superior analgesia. These studies suggest that ASB E. coli represent novel LBPs for UTI symptoms. PMID:25405579

Yaggie, Ryan E.; Schaeffer, Anthony J.; Klumpp, David J.

2014-01-01

151

Biosynthesis of 4-Aminobenzoate in Escherichia coli  

PubMed Central

Two different mutations (pabA and pabB) affecting 4-aminobenzoate biosynthesis were obtained in strains of Escherichia coli lacking chorismate mutase and anthranilate synthetase activity, thus allowing study of the pathway of biosynthesis of 4-aminobenzoate by use of cell extracts of strains carrying the pab mutations. Two components with approximate molecular weights of 9,000 (component A) and 48,000 (component B) are concerned in the biosynthesis of 4-aminobenzoate from chorismate by E. coli. No diffusible intermediate compound could be detected. PMID:4914080

Huang, Minta; Gibson, F.

1970-01-01

152

Rotation of Escherichia coli F 1ATPase  

Microsoft Academic Search

By applying the same method used for F1-ATPase (TF1) from thermophilic Bacillus PS3 (Noji, H., Yasuda, R., Yoshida, M., and Kinosita, K., Jr. (1997) Nature 386, 299–302), we observed ATP-driven rotation of a fluorescent actin filament attached to the ? subunit in Escherichia coli F1-ATPase. The torque value and the direction of the rotation were the same as those observed

Hiroyuki Noji; Katrin Häsler; Wolfgang Junge; Kazuhiko Kinosita; Masasuke Yoshida; Siegfried Engelbrecht

1999-01-01

153

Energetics of sodium efflux from Escherichia coli.  

PubMed

When energy-starved cells of Escherichia coli were passively loaded with 22Na+, efflux of sodium could be initiated by addition of a source of metabolic energy. Conditions were established where the source of energy was phosphate bond energy, an electrochemical proton gradient, or both. Only an electrochemical proton gradient was required for efflux from intact cells. These results are consistent with secondary exchange of Na+ for H+ catalyzed by a sodium/proton antiporter. PMID:6322694

Borbolla, M G; Rosen, B P

1984-02-15

154

Escherichia coli Biofilms Have an Organized and Complex Extracellular Matrix Structure  

PubMed Central

ABSTRACT Bacterial biofilms are ubiquitous in nature, and their resilience is derived in part from a complex extracellular matrix that can be tailored to meet environmental demands. Although common developmental stages leading to biofilm formation have been described, how the extracellular components are organized to allow three-dimensional biofilm development is not well understood. Here we show that uropathogenic Escherichia coli (UPEC) strains produce a biofilm with a highly ordered and complex extracellular matrix (ECM). We used electron microscopy (EM) techniques to image floating biofilms (pellicles) formed by UPEC. EM revealed intricately constructed substructures within the ECM that encase individual, spatially segregated bacteria with a distinctive morphology. Mutational and biochemical analyses of these biofilms confirmed curli as a major matrix component and revealed important roles for cellulose, flagella, and type 1 pili in pellicle integrity and ECM infrastructure. Collectively, the findings of this study elucidated that UPEC pellicles have a highly organized ultrastructure that varies spatially across the multicellular community. PMID:24023384

Hung, Chia; Zhou, Yizhou; Pinkner, Jerome S.; Dodson, Karen W.; Crowley, Jan R.; Heuser, John; Chapman, Matthew R.; Hadjifrangiskou, Maria; Henderson, Jeffrey P.; Hultgren, Scott J.

2013-01-01

155

Genetic relationships among Escherichia coli isolates causing urinary tract infections in humans and animals.  

PubMed Central

Genetic variation in isolates of Escherichia coli obtained mostly from urinary tract infections in humans and domesticated animals (dogs and cats) was assessed for 16 enzymes using multilocus enzyme electrophoresis to characterize chromosomal genotypes. A total of 148 isolates comprised 63 distinct electrophoretic types (ETs) and about half of the isolates belonged to one of 9 common ETs. A bootstrap analysis of genetic distance between ETs revealed three significant groups of strains. Variation in allele frequencies among groups accounted for 40% of the total genetic diversity. The majority of the common ETs fell into a major cluster of closely related strains. The recovery of multiple isolates of the same electrophoretic types and serotypes from unassociated hosts suggests that these bacteria represent uropathogenic clones that are widely disseminated in humans and animals. PMID:2645153

Whittam, T. S.; Wolfe, M. L.; Wilson, R. A.

1989-01-01

156

Action of sodium deoxycholate on Escherichia coli  

SciTech Connect

Sodium deoxycholate is used in a number of bacteriological media for the isolation and classification of gram-negative bacteria from food and the environment. Initial experiments to study the effect of deoxycholate on the growth parameters of Escherichia coli showed an increase in the lag time constant and generation time and a decrease in the growth rate constant total cell yield of this microorganisms. Cell fractionation studies indicated that sodium deoxycholate at levels used in bacteriological media interferes with the incorporation of (U-/sup 14/C)glucose into the cold-trichloroacetic acid-soluble, ethanol-soluble, and trypsin-soluble cellular fractions of E. coli. Finally, sodium deoxycholate interfered with the flagellation and motility of Proteus mirabilis and E. coli. It would appear then that further improvement of the deoxycholate medium may be in order.

D'Mello, A.; Yotis, W.W.

1987-08-01

157

Biotyping of Escherichia coli in microwell plates.  

PubMed

A simple, inexpensive scheme of eight tests for biotyping strains of Escherichia coli in microwell plates is described. The tests comprise primary tests for the fermentation of raffinose, sorbose, ornithine, dulcitol and 2-deoxy-D-ribose, and secondary tests for rhamnose fermentation, lysine decarboxylation and motility. Among a collection of 75 clinical isolates of Esch. coli from 12 patients, 18 full biotypes designated according to their positive and negative reactions in the eight tests were distinguished. These biotypes gave an indication of the natural history of patients' infections. Because it provides excellent and reliable type discrimination, biotyping can be used in a combination with other typing techniques to resolve local epidemiological problems involving Esch. coli. PMID:8527993

Crichton, P B; Taylor, A

1995-09-01

158

Molecular epidemiology of Escherichia coli O25b-ST131 isolates causing community-acquired UTIs in Mexico.  

PubMed

Escherichia coli is a common uropathogen causing community-acquired urinary tract infections (UTIs). Out of 4735 E. coli community-acquired UTIs, 10.2% were extended spectrum ?-lactamases (ESBL)-producing. The identified ESBL types were CTX-M-15 (96.4%), SHV-2a (3%), and TLA-1 (1%). Of the isolates, 94.6% tested positive for plasmid-mediated quinolone resistance (PMQR) genes (aac(6')-lb-cr [92.1%] and qepA1 [7%] and for qnr-determinants [3.5%]). E. coli O25b-ST131 was identified in 25% of the isolates that harbor a non-conjugative 160-kb plasmid (IncFIA) containing the CTX-M-15, and all of these isolates were found to contain PMQR genes. This work can be useful in modeling the potential impact that may have on community-acquired UTIs in Mexico. PMID:23774006

Reyna-Flores, Fernando; Barrios, Humberto; Garza-Ramos, Ulises; Sánchez-Pérez, Alejandro; Rojas-Moreno, Teresa; Uribe-Salas, Felipe J; Fagundo-Sierra, Reynero; Silva-Sanchez, Jesus

2013-07-01

159

Salmonella typhimurium intercepts Escherichia coli signaling to enhance antibiotic tolerance  

E-print Network

Salmonella typhimurium intercepts Escherichia coli signaling to enhance antibiotic tolerance Nicole Salmonella typhimurium increases its antibiotic toler- ance in response to indole, even though S. typhimurium

Collins, James J.

160

Survival of multi-drug resistant enteropathogenic Escherichia coli and Salmonella paratyphi in Vembanadu  

E-print Network

Survival of multi-drug resistant enteropathogenic Escherichia coli and Salmonella paratyphi the survival response of multi-drug resistant enteropathogenic Escherichia coli and Salmonella paratyphi- otypes of Escherichia coli, Salmonella enterica typhi and paratyphi are highly endemic to India

Mazumder, Asit

161

Biodegradation of Aromatic Compounds by Escherichia coli  

PubMed Central

Although Escherichia coli has long been recognized as the best-understood living organism, little was known about its abilities to use aromatic compounds as sole carbon and energy sources. This review gives an extensive overview of the current knowledge of the catabolism of aromatic compounds by E. coli. After giving a general overview of the aromatic compounds that E. coli strains encounter and mineralize in the different habitats that they colonize, we provide an up-to-date status report on the genes and proteins involved in the catabolism of such compounds, namely, several aromatic acids (phenylacetic acid, 3- and 4-hydroxyphenylacetic acid, phenylpropionic acid, 3-hydroxyphenylpropionic acid, and 3-hydroxycinnamic acid) and amines (phenylethylamine, tyramine, and dopamine). Other enzymatic activities acting on aromatic compounds in E. coli are also reviewed and evaluated. The review also reflects the present impact of genomic research and how the analysis of the whole E. coli genome reveals novel aromatic catabolic functions. Moreover, evolutionary considerations derived from sequence comparisons between the aromatic catabolic clusters of E. coli and homologous clusters from an increasing number of bacteria are also discussed. The recent progress in the understanding of the fundamentals that govern the degradation of aromatic compounds in E. coli makes this bacterium a very useful model system to decipher biochemical, genetic, evolutionary, and ecological aspects of the catabolism of such compounds. In the last part of the review, we discuss strategies and concepts to metabolically engineer E. coli to suit specific needs for biodegradation and biotransformation of aromatics and we provide several examples based on selected studies. Finally, conclusions derived from this review may serve as a lead for future research and applications. PMID:11729263

Díaz, Eduardo; Ferrández, Abel; Prieto, María A.; García, José L.

2001-01-01

162

Comparison of host response mechanisms evoked by extended spectrum beta lactamase (ESBL)- and non-ESBL-producing uropathogenic E. coli  

PubMed Central

Background Infections caused by extended spectrum beta-lactamases (ESBL)-producing bacteria have been emerging worldwide and the majority of ESBL-producing E. coli strains are isolated from patients with urinary tracts infections. The purpose of this study was to compare the host-response mechanisms in human polymorphonucleated leukocytes (PMN) and renal epithelial cells when stimulated by ESBL- or non-ESBL-producing uropathogenic E. coli (UPEC) isolates. The host-pathogen interaction of these ESBL-producing strains in the urinary tract is not well studied. Results The ability of ESBL strains to evoke ROS-production from PMN cells was significantly higher than that of the non-ESBL strains. The growth of ESBL strains was slightly suppressed in the presence of PMN compared to non-ESBL strains after 30 min and 2 h, but the opposite was observed after 5 and 6 h. The number of migrating PMN was significantly higher in response to ESBL strains compared to non-ESBL strains. Stimulation of A498 cells with ESBL strains elicited lower production of IL-6 and IL-8 compared to non-ESBL strains. Conclusion Significant differences in host-response mechanisms were identified when host cells were stimulated by ESBL- or non-ESBL producing strains. The obtained results on the early interactions of ESBL-producing strains with the host immune system may provide valuable information for management of these infections. PMID:24059789

2013-01-01

163

Escherichia coli fimbriae recognizing sialyl galactosides.  

PubMed Central

Fimbriae recognizing sialyl galactosides (S fimbriae) were purified from an Escherichia coli strain. The S fimbriae were morphologically identical to type 1 and P fimbriae of E. coli and showed a hemagglutination that was abolished when erythrocytes were treated with neuraminidase. Hemagglutination by the purified fimbriae was inhibited by orosomucoid but not by its desialylated derivative. Of the oligosaccharides tested, sialyl-(alpha 2-3)-lactose and sialyl-(alpha 2-3)-N-acetyllactosamine had the strongest inhibitory activities. It was concluded that S fimbriae have the strongest affinity for (alpha 2-3)-linked sialyl galactosides. In the enzyme-linked immunosorbent assay, the hyperimmune serum to the S fimbriae reacted strongly with the homologous antigen but not with type 1, P, or nonhemagglutinating KS71C fimbriae of E. coli. Analogously, the hyperimmune sera to the other E. coli fimbriae did not react with the purified S fimbriae. The immunoprecipitation assay showed that S fimbriae on different E. coli serotypes shared immunological cross-reactivity. Images PMID:6146600

Korhonen, T K; Väisänen-Rhen, V; Rhen, M; Pere, A; Parkkinen, J; Finne, J

1984-01-01

164

Escherichia coli fimbriae recognizing sialyl galactosides.  

PubMed

Fimbriae recognizing sialyl galactosides (S fimbriae) were purified from an Escherichia coli strain. The S fimbriae were morphologically identical to type 1 and P fimbriae of E. coli and showed a hemagglutination that was abolished when erythrocytes were treated with neuraminidase. Hemagglutination by the purified fimbriae was inhibited by orosomucoid but not by its desialylated derivative. Of the oligosaccharides tested, sialyl-(alpha 2-3)-lactose and sialyl-(alpha 2-3)-N-acetyllactosamine had the strongest inhibitory activities. It was concluded that S fimbriae have the strongest affinity for (alpha 2-3)-linked sialyl galactosides. In the enzyme-linked immunosorbent assay, the hyperimmune serum to the S fimbriae reacted strongly with the homologous antigen but not with type 1, P, or nonhemagglutinating KS71C fimbriae of E. coli. Analogously, the hyperimmune sera to the other E. coli fimbriae did not react with the purified S fimbriae. The immunoprecipitation assay showed that S fimbriae on different E. coli serotypes shared immunological cross-reactivity. PMID:6146600

Korhonen, T K; Väisänen-Rhen, V; Rhen, M; Pere, A; Parkkinen, J; Finne, J

1984-08-01

165

Core and Panmetabolism in Escherichia coli? †  

PubMed Central

Escherichia coli exhibits a wide range of lifestyles encompassing commensalism and various pathogenic behaviors which its highly dynamic genome contributes to develop. How environmental and host factors shape the genetic structure of E. coli strains remains, however, largely unknown. Following a previous study of E. coli genomic diversity, we investigated its diversity at the metabolic level by building and analyzing the genome-scale metabolic networks of 29 E. coli strains (8 commensal and 21 pathogenic strains, including 6 Shigella strains). Using a tailor-made reconstruction strategy, we significantly improved the completeness and accuracy of the metabolic networks over default automatic reconstruction processes. Among the 1,545 reactions forming E. coli panmetabolism, 885 reactions were common to all strains. This high proportion of core reactions (57%) was found to be in sharp contrast to the low proportion (13%) of core genes in the E. coli pangenome, suggesting less diversity of metabolic functions compared to that of all gene functions. Core reactions were significantly overrepresented among biosynthetic reactions compared to the more variable degradation processes. Differences between metabolic networks were found to follow E. coli phylogeny rather than pathogenic phenotypes, except for Shigella networks, which were significantly more distant from the others. This suggests that most metabolic changes in non-Shigella strains were not driven by their pathogenic phenotypes. Using a supervised method, we were yet able to identify small sets of reactions related to pathogenicity or commensalism. The quality of our reconstructed networks also makes them reliable bases for building metabolic models. PMID:21239590

Vieira, Gilles; Sabarly, Victor; Bourguignon, Pierre-Yves; Durot, Maxime; Le Fèvre, François; Mornico, Damien; Vallenet, David; Bouvet, Odile; Denamur, Erick; Schachter, Vincent; Médigue, Claudine

2011-01-01

166

Role of Deoxyribose Catabolism in Colonization of the Murine Intestine by Pathogenic Escherichia coli Strains ?  

PubMed Central

We previously suggested that the ability to metabolize deoxyribose, a phenotype encoded by the deoK operon, is associated with the pathogenic potential of Escherichia coli strains. Carbohydrate metabolism is thought to provide the nutritional support required for E. coli to colonize the intestine. We therefore investigated the role of deoxyribose catabolism in the colonization of the gut, which acts as a reservoir, by pathogenic E. coli strains. Molecular and biochemical characterization of 1,221 E. coli clones from various collections showed this biochemical trait to be common in the E. coli species (33.6%). However, multivariate analysis evidenced a higher prevalence of sugar-metabolizing E. coli clones in the stools of patients from countries in which intestinal diseases are endemic. Diarrhea processes frequently involve the destruction of intestinal epithelia, so it is plausible that such clones may be positively selected for in intestines containing abundant DNA, and consequently deoxyribose. Statistical analysis also indicated that symptomatic clinical disorders and the presence of virulence factors specific to extraintestinal pathogenic E. coli were significantly associated with an increased risk of biological samples and clones testing positive for deoxyribose. Using the streptomycin-treated-mouse model of intestinal colonization, we demonstrated the involvement of the deoK operon in gut colonization by two pathogenic isolates (one enteroaggregative and one uropathogenic strain). These results, indicating that deoxyribose availability promotes pathogenic E. coli growth during host colonization, suggest that the acquisition of this trait may be an evolutionary step enabling these pathogens to colonize and persist in the mammalian intestine. PMID:19168744

Martinez-Jéhanne, Vanessa; du Merle, Laurence; Bernier-Fébreau, Christine; Usein, Codruta; Gassama-Sow, Amy; Wane, Abdul-Aziz; Gouali, Malika; Damian, Maria; Aïdara-Kane, Awa; Germani, Yves; Fontanet, Arnaud; Coddeville, Bernadette; Guérardel, Yann; Le Bouguénec, Chantal

2009-01-01

167

The antibacterial effect of nitric oxide against ESBL-producing uropathogenic E. coli is improved by combination with miconazole and polymyxin B nonapeptide  

PubMed Central

Background Nitric oxide (NO) is produced as part of the host immune response to bacterial infections, including urinary tract infections. The enzyme flavohemoglobin, coded by the hmp gene, is involved in protecting bacterial cells from the toxic effects of NO and represents a potentially interesting target for development of novel treatment concepts against resistant uropathogenic bacteria. The aim of the present study was to investigate if the in vitro antibacterial effects of NO can be enhanced by pharmacological modulation of the enzyme flavohemoglobin. Results Four clinical isolates of multidrug-resistant extended-spectrum ?-lactamase (ESBL)-producing uropathogenic E. coli were included in the study. It was shown that the NO-donor substance DETA/NO, but not inactivated DETA/NO, caused an initial growth inhibition with regrowth noted after 8?h of exposure. An hmp-deficient strain showed a prolonged growth inhibition in response to DETA/NO compared to the wild type. The imidazole antibiotic miconazole, that has been shown to inhibit bacterial flavohemoglobin activity, prolonged the DETA/NO-evoked growth inhibition. When miconazole was combined with polymyxin B nonapeptide (PMBN), in order to increase the bacterial wall permeability, DETA/NO caused a prolonged bacteriostatic response that lasted for up to 24?h. Conclusion An NO-donor in combination with miconazole and PMBN showed enhanced antimicrobial effects and proved effective against multidrug-resistant ESBL-producing uropathogenic E. coli. PMID:24629000

2014-01-01

168

Gas signatures from Escherichia coli and Escherichia coli-inoculated human whole blood  

PubMed Central

Background The gaseous headspace above naïve Escherichia Coli (E. coli) cultures and whole human blood inoculated with E. coli were collected and analyzed for the presence of trace gases that may have the potential to be used as novel, non-invasive markers of infectious disease. Methods The naïve E. coli culture, LB broth, and human whole blood or E. coli inoculated whole blood were incubated in hermetically sealable glass bioreactors at 37°C for 24 hrs. LB broth and whole human blood were used as controls for background volatile organic compounds (VOCs). The headspace gases were collected after incubation and analyzed using a gas chromatographic system with multiple column/detector combinations. Results Six VOCs were observed to be produced by E. coli-infected whole blood while there existed nearly zero to relatively negligible amounts of these gases in the whole blood alone, LB broth, or E. coli-inoculated LB broth. These VOCs included dimethyl sulfide (DMS), carbon disulfide (CS2), ethanol, acetaldehyde, methyl butanoate, and an unidentified gas S. In contrast, there were several VOCs significantly elevated in the headspace above the E. coli in LB broth, but not present in the E. coli/blood mixture. These VOCs included dimethyl disulfide (DMDS), dimethyl trisulfide (DMTS), methyl propanoate, 1-propanol, methylcyclohexane, and unidentified gases R2 and Q. Conclusions This study demonstrates 1) that cultivated E. coli in LB broth produce distinct gas profiles, 2) for the first time, the ability to modify E. coli-specific gas profiles by the addition of whole human blood, and 3) that E. coli-human whole blood interactions present different gas emission profiles that have the potential to be used as non-invasive volatile biomarkers of E. coli infection. PMID:23842518

2013-01-01

169

Engineering Desiccation Tolerance in Escherichia coli  

PubMed Central

Recombinant sucrose-6-phosphate synthase (SpsA) was synthesized in Escherichia coli BL21DE3 by using the spsA gene of the cyanobacterium Synechocystis sp. strain PCC 6803. Transformants exhibited a 10,000-fold increase in survival compared to wild-type cells following either freeze-drying, air drying, or desiccation over phosphorus pentoxide. The phase transition temperatures and vibration frequencies (P?O stretch) in phospholipids suggested that sucrose maintained membrane fluidity during cell dehydration. PMID:10742260

Billi, Daniela; Wright, Deborah J.; Helm, Richard F.; Prickett, Todd; Potts, Malcolm; Crowe, John H.

2000-01-01

170

Engineering ethanologenic Escherichia coli for levoglucosan utilization.  

PubMed

Levoglucosan is a major product of biomass pyrolysis. While this pyrolyzed biomass, also known as bio-oil, contains sugars that are an attractive fermentation substrate, commonly-used biocatalysts, such as Escherichia coli, lack the ability to metabolize this anhydrosugar. It has previously been shown that recombinant expression of the levoglucosan kinase enzyme enables use of levoglucosan as carbon and energy source. Here, ethanologenic E. coli KO11 was engineered for levoglucosan utilization by recombinant expression of levoglucosan kinase from Lipomyces starkeyi. Our engineering strategy uses a codon-optimized gene that has been chromosomally integrated within the pyruvate to ethanol (PET) operon and does not require additional antibiotics or inducers. Not only does this engineered strain use levoglucosan as sole carbon source, but it also ferments levoglucosan to ethanol. This work demonstrates that existing biocatalysts can be easily modified for levoglucosan utilization. PMID:21719279

Layton, Donovan S; Ajjarapu, Avanthi; Choi, Dong Won; Jarboe, Laura R

2011-09-01

171

Automatic Tracking of Escherichia Coli Bacteria , Shahid Khan2 3  

E-print Network

of choice for elucidation of the design principles of transmembrane and intracellular signal transductionAutomatic Tracking of Escherichia Coli Bacteria Jun Xie1 , Shahid Khan2 3 , and Mubarak Shah4 1. In this paper, we present an automatic method for estimating the tra- jectories of Escherichia coli bacteria

Central Florida, University of

172

Production of glycoprotein vaccines in Escherichia coli  

PubMed Central

Background Conjugate vaccines in which polysaccharide antigens are covalently linked to carrier proteins belong to the most effective and safest vaccines against bacterial pathogens. State-of-the art production of conjugate vaccines using chemical methods is a laborious, multi-step process. In vivo enzymatic coupling using the general glycosylation pathway of Campylobacter jejuni in recombinant Escherichia coli has been suggested as a simpler method for producing conjugate vaccines. In this study we describe the in vivo biosynthesis of two novel conjugate vaccine candidates against Shigella dysenteriae type 1, an important bacterial pathogen causing severe gastro-intestinal disease states mainly in developing countries. Results Two different periplasmic carrier proteins, AcrA from C. jejuni and a toxoid form of Pseudomonas aeruginosa exotoxin were glycosylated with Shigella O antigens in E. coli. Starting from shake flask cultivation in standard complex medium a lab-scale fed-batch process was developed for glycoconjugate production. It was found that efficiency of glycosylation but not carrier protein expression was highly susceptible to the physiological state at induction. After induction glycoconjugates generally appeared later than unglycosylated carrier protein, suggesting that glycosylation was the rate-limiting step for synthesis of conjugate vaccines in E. coli. Glycoconjugate synthesis, in particular expression of oligosaccharyltransferase PglB, strongly inhibited growth of E. coli cells after induction, making it necessary to separate biomass growth and recombinant protein expression phases. With a simple pulse and linear feed strategy and the use of semi-defined glycerol medium, volumetric glycoconjugate yield was increased 30 to 50-fold. Conclusions The presented data demonstrate that glycosylated proteins can be produced in recombinant E. coli at a larger scale. The described methodologies constitute an important step towards cost-effective in vivo production of conjugate vaccines, which in future may be used for combating severe infectious diseases, particularly in developing countries. PMID:20701771

2010-01-01

173

Sources of Escherichia coli in a Coastal Subtropical Environment  

Microsoft Academic Search

Sources of Escherichia coli in a coastal waterway located in Ft. Lauderdale, Fla., were evaluated. The study consisted of an extensive program of field measurements designed to capture spatial and temporal variations in E. coli concentrations as well as experiments conducted under laboratory-controlled conditions. E. coli from environmental samples was enumerated by using a defined substrate technology (Colilert-18). Field sampling

HELENA M. SOLO-GABRIELE; MELINDA A. WOLFERT; TIMOTHY R. DESMARAIS; CAROL J. PALMER

2000-01-01

174

Engineering a Reduced Escherichia coli Genome  

PubMed Central

Our goal is to construct an improved Escherichia coli to serve both as a better model organism and as a more useful technological tool for genome science. We developed techniques for precise genomic surgery and applied them to deleting the largest K-islands of E. coli, identified by comparative genomics as recent horizontal acquisitions to the genome. They are loaded with cryptic prophages, transposons, damaged genes, and genes of unknown function. Our method leaves no scars or markers behind and can be applied sequentially. Twelve K-islands were successfully deleted, resulting in an 8.1% reduced genome size, a 9.3% reduction of gene count, and elimination of 24 of the 44 transposable elements of E. coli. These are particularly detrimental because they can mutagenize the genome or transpose into clones being propagated for sequencing, as happened in 18 places of the draft human genome sequence. We found no change in the growth rate on minimal medium, confirming the nonessential nature of these islands. This demonstration of feasibility opens the way for constructing a maximally reduced strain, which will provide a clean background for functional genomics studies, a more efficient background for use in biotechnology applications, and a unique tool for studies of genome stability and evolution. [Sequence data described in this paper have been submitted to the DNA Data Bank of Japan, European Molecular Biology Laboratory, and GenBank databases under accession nos. AF402780, AF402779, and AF406953, respectively.] PMID:11932248

Kolisnychenko, Vitaliy; Plunkett, Guy; Herring, Christopher D.; Fehér, Tamás; Pósfai, János; Blattner, Frederick R.; Pósfai, György

2002-01-01

175

Autophosphorylation of phosphoglucosamine mutase from Escherichia coli.  

PubMed

Phosphoglucosamine mutase (GlmM) catalyzes the formation of glucosamine-1-phosphate from glucosamine-6-phosphate, an essential step in the pathway for UDP-N-acetylglucosamine biosynthesis in bacteria. This enzyme must be phosphorylated to be active and acts according to a ping-pong mechanism involving glucosamine-1, 6-diphosphate as an intermediate (L. Jolly, P. Ferrari, D. Blanot, J. van Heijenoort, F. Fassy, and D. Mengin-Lecreulx, Eur. J. Biochem. 262:202-210, 1999). However, the process by which the initial phosphorylation of the enzyme is achieved in vivo remains unknown. Here we show that the phosphoglucosamine mutase from Escherichia coli autophosphorylates in vitro in the presence of [(32)P]ATP. The same is observed with phosphoglucosamine mutases from other bacterial species, yeast N-acetylglucosamine-phosphate mutase, and rabbit muscle phosphoglucomutase. Labeling of the E. coli GlmM enzyme with [(32)P]ATP requires the presence of a divalent cation, and the label is subsequently lost when the enzyme is incubated with either of its substrates. Analysis of enzyme phosphorylation by high-pressure liquid chromatography and coupled mass spectrometry confirms that only one phosphate has been covalently linked to the enzyme. Only phosphoserine could be detected after acid hydrolysis of the labeled protein, and site-directed mutagenesis of serine residues located in or near the active site identifies the serine residue at position 102 as the site of autophosphorylation of E. coli GlmM. PMID:10671448

Jolly, L; Pompeo, F; van Heijenoort, J; Fassy, F; Mengin-Lecreulx, D

2000-03-01

176

Characterization of molybdenum cofactor from Escherichia coli.  

PubMed Central

Molybdenum cofactor activity was found in the soluble fraction of cell-free extracts of Escherichia coli grown aerobically in media supplemented with molybdate. Cofactor was detected by its ability to complement the nitrate reductase-deficient mutant of Neurospora crossa, nit-1, resulting in the vitro formation of nitrate reductase activity. Acid treatment of E. coli extracts was not required for release of cofactor activity. Cofactor was able to diffuse through a membrane of nominal 2,000-molecular-weight cutoff and was insensitive to trypsin. The cofactor was associated with a carrier molecule (approximately 40,000 daltons) during gel filtration and sucrose gradient centrifugation, but was easily removed from the carrier by dialysis. The carrier molecule protected the cofactor from inactivation by heat or oxygen. E. coli grown in molybdenum-free media, without and with tungsten, synthesized a metal-free "empty" cofactor and its tungsten analog, respectively, both of which were subsequently activated by the addition of molybdate. Empty and tungsten-containing cofactor complemented the nitrate reductase subunits in the nit-1 extract, forming inactive, but intact, 7.9S nitrate reductase. Addition of molybdate to the enzyme complemented in this manner restored nitrate reductase activity. PMID:387715

Amy, N K; Rajagopalan, K V

1979-01-01

177

Nonlethal adherence to human neutrophils mediated by Dr antigen-specific adhesins of Escherichia coli.  

PubMed

Uropathogenic Escherichia coli strains express a variety of adhesins, including members of the Dr adhesin family such as the Dr hemagglutinin, AFAI, and AFAIII. Certain E. coli adhesins (e.g., type 1 and S fimbriae) are known to mediate adherence to human polymorphonuclear leukocytes (PMNs). The receptor on erythrocytes for Dr family adhesins, decay accelerating factor, is also present on PMNs. To determine whether Dr family adhesins mediate adherence to PMNs and to characterize the specificity and consequences of such adherence, we studied agglutination of PMNs and adherence to PMNs by recombinant E. coli strains expressing various mannose-resistant or mannose-sensitive adhesins, in the presence or absence of inhibitors of adherence. Dr family adhesins, like type 1 fimbriae, mediated concentration-dependent adherence to PMNs. Adherence to PMNs was mannose sensitive for type 1 fimbriae but mannose resistant for Dr family adhesins. Chloramphenicol inhibited PMN adherence for the Dr hemagglutinin with the same potency as that with which it inhibited hemagglutination, but it was inactive against PMN adherence and hemagglutination mediated by other members of the Dr adhesin family. In contrast to PMN adherence mediated by type 1 fimbriae, adherence mediated by the Dr hemagglutinin did not lead to significantly increased bacterial killing. These data suggest that Dr family adhesins mediate a novel pattern of adherence to PMNs, probably by recognizing decay accelerating factor, with minimal consequent bacterial killing. PMID:7806371

Johnson, J R; Skubitz, K M; Nowicki, B J; Jacques-Palaz, K; Rakita, R M

1995-01-01

178

Vibrio parahaemolyticus, enterotoxigenic Escherichia coli, enterohemorrhagic Escherichia coli and Vibrio cholerae  

PubMed Central

This review highlighted the following: (i) pathogenic mechanism of the thermostable direct hemolysin produced by Vibrio parahaemolyticus, especially on its cardiotoxicity, (ii) heat-labile and heat-stable enterotoxins produced by enterotoxigenic Escherichia coli, especially structure–activity relationship of heat-stable enterotoxin, (iii) RNA N-glycosidase activity of Vero toxins (VT1 and VT2) produced by enterohemorrhagic Escherichia coli O157:H7, (iv) discovery of Vibrio cholerae O139, (v) isolation of new variant of Vibrio cholerae O1 El Tor that carries classical ctxB, and production of high concentration of cholera toxin by these strains, and (vi) conversion of viable but nonculturable (VBNC) Vibrio cholerae to culturable state by co-culture with eukaryotic cells. PMID:21233598

TAKEDA, Yoshifumi

2011-01-01

179

Indole transport across Escherichia coli membranes.  

PubMed

Indole has many, diverse roles in bacterial signaling. It regulates the transition from exponential to stationary phase, it is involved in the control of plasmid stability, and it influences biofilm formation, virulence, and stress responses (including antibiotic resistance). Its role is not restricted to bacteria, and recently it has been shown to include mutually beneficial signaling between enteric bacteria and their mammalian hosts. In many respects indole behaves like the signaling component of a quorum-sensing system. Indole synthesized within the producer bacterium is exported into the surroundings where its accumulation is detected by sensitive cells. A view often repeated in the literature is that in Escherichia coli the AcrEF-TolC and Mtr transporter proteins are involved in the export and import, respectively, of indole. However, the evidence for their involvement is indirect, and it has been known for a long time that indole can pass directly through a lipid bilayer. We have combined in vivo and in vitro approaches to examine the relative importance of protein-mediated transport and direct passage across the E. coli membrane. We conclude that the movement of indole across the E. coli membrane under normal physiological conditions is independent of AcrEF-TolC and Mtr. Furthermore, direct observation of individual liposomes shows that indole can rapidly cross an E. coli lipid membrane without the aid of any proteinaceous transporter. These observations not only enhance our understanding of indole signaling in bacteria but also provide a simple explanation for the ability of indole to signal between biological kingdoms. PMID:21296966

Piñero-Fernandez, S; Chimerel, C; Keyser, U F; Summers, D K

2011-04-01

180

Enteroaggregative Escherichia coli O78:H10, the Cause of an Outbreak of Urinary Tract Infection  

PubMed Central

In 1991, multiresistant Escherichia coli O78:H10 strains caused an outbreak of urinary tract infections in Copenhagen, Denmark. The phylogenetic origin, clonal background, and virulence characteristics of the outbreak isolates, and their relationship to nonoutbreak O78:H10 strains according to these traits and resistance profiles, are unknown. Accordingly, we extensively characterized 51 archived E. coli O78:H10 isolates (48 human isolates from seven countries, including 19 Copenhagen outbreak isolates, and 1 each of calf, avian, and unknown-source isolates), collected from 1956 through 2000. E. coli O78:H10 was clonally heterogeneous, comprising one dominant clonal group (61% of isolates, including all 19 outbreak isolates) from ST10 (phylogenetic group A) plus several minor clonal groups (phylogenetic groups A and D). All ST10 isolates, versus 25% of non-ST10 isolates, were identified by molecular methods as enteroaggregative E. coli (EAEC) (P < 0.001). Genes present in >90% of outbreak isolates included fimH (type 1 fimbriae; ubiquitous in E. coli); fyuA, traT, and iutA (associated with extraintestinal pathogenic E. coli [ExPEC]); and sat, pic, aatA, aggR, aggA, ORF61, aaiC, aap, and ORF3 (associated with EAEC). An outbreak isolate was lethal in a murine subcutaneous sepsis model and exhibited characteristic EAEC “stacked brick” adherence to cultured epithelial cells. Thus, the 1991 Copenhagen outbreak was caused by a tight, non-animal-associated subset within a broadly disseminated O78:H10 clonal group (ST10; phylogenetic group A), members of which exhibit both ExPEC and EAEC characteristics, whereas O78:H10 isolates overall are phylogenetically diverse. Whether ST10 O78:H10 EAEC strains are both uropathogenic and diarrheagenic warrants further investigation. PMID:22972830

Scheutz, Flemming; Andersen, Rebecca L.; Menard, Megan; Boisen, Nadia; Johnston, Brian; Hansen, Dennis S.; Krogfelt, Karen A.; Nataro, James P.; Johnson, James R.

2012-01-01

181

Virulence factors and O groups of Escherichia coli isolates from patients with acute pyelonephritis, cystitis and asymptomatic bacteriuria.  

PubMed

The relationship between the presence of bacterial virulence factors and the severity of urinary tract infection (UTI) was analized in this study. The production of alpha-hemolysin (Hly), the expression of P-fimbriae and the mannose-resistant hemagglutination (MRHA) type IVa (associated with the presence of P-fimbriae), were all detected more frequently in Escherichia coli strains from acute pyelonephritis than in strains isolated from cystitis and asymptomatic bacteriuria. In contrast, the production of cytotoxic necrotizing factor type 1 (CNF1) and the expression of MRHA types III and IVb were distributed uniformly between strains causing different clinical categories of UTI. Thus 88% of the E. coli strains from acute pyelonephritis showed some of the virulence factors investigated in this study, whereas only 60% (p < 0.01) and 56% (p < 0.01) repectively of the strains isolated from cystitis and asymptomatic bacteriuria possessed virulence factors. There were no significant differences in the prevalence of virulence properties between strains isolated from patients with or without complicating factors. Only 16% (p < 0.001) of the fecal isolates from healthy individuals showed virulence factors. The virulence factors were concentrated in strains belonging to 10 (O1, O2, O4, O6, O7, O14, O18, O22, O75 and O83) of the 12 serogroups most frequently detected in uropathogenic E. coli strains. The majority of uropathogenic O4, O6, O14, O22, O75 and O83 E.coli strains were Hly+CNF1+ and expressed P-fimbriae or MRHA type III, whereas the strains of serogroup O18 were Hly+CNF1- and P-fimbriated. Among O1 and O7 strains we found Hly-CNF1-strains that expressed P-fimbriae. Among O2 strains we found Hly+CNF1+ strains that expressed P-fimbriae or MRHA type III and other Hly-CNF1-strains that possessed P-fimbriae. We conclude that E.coli strains isolated from pyelonephritis show virulence factors more frequently than those from cystitis and asymptomatic bacteriuria, and that strains that cause urinary tract infections in Spain belong to the same serogroups as uropathogenic E.coli isolated in other areas of the world. Our results support the special pathogenicity theory and suggest that many cases of serious urogenital disease may be caused by a limited number of P-fimbriated E.coli strains that usually produce alpha-hemolysin. PMID:8817199

Blanco, M; Blanco, J E; Alonso, M P; Blanco, J

1996-04-01

182

Dispensability of Escherichia coli's latent pathways  

E-print Network

Gene-knockout experiments on single-cell organisms have established that expression of a substantial fraction of genes is not needed for optimal growth. This problem acquired a new dimension with the recent discovery that environmental and genetic perturbations of the bacterium Escherichia coli are followed by the temporary activation of a large number of latent metabolic pathways, which suggests the hypothesis that temporarily activated reactions impact growth and hence facilitate adaptation in the presence of perturbations. Here we test this hypothesis computationally and find, surprisingly, that the availability of latent pathways consistently offers no growth advantage, and tends in fact to inhibit growth after genetic perturbations. This is shown to be true even for latent pathways with a known function in alternate conditions, thus extending the significance of this adverse effect beyond apparently nonessential genes. These findings raise the possibility that latent pathway activation is in fact derivat...

Cornelius, Sean P; Motter, Adilson E; 10.1073/pnas.1009772108

2011-01-01

183

Mechanism of Escherichia coli Resistance to Pyrrhocoricin  

PubMed Central

Due to their lack of toxicity to mammalian cells and good serum stability, proline-rich antimicrobial peptides (PR-AMPs) have been proposed as promising candidates for the treatment of infections caused by antimicrobial-resistant bacterial pathogens. It has been hypothesized that these peptides act on multiple targets within bacterial cells, and therefore the likelihood of the emergence of resistance was considered to be low. Here, we show that spontaneous Escherichia coli mutants resistant to pyrrhocoricin arise at a frequency of approximately 6 × 10?7. Multiple independently derived mutants all contained a deletion in a nonessential gene that encodes the putative peptide uptake permease SbmA. Sensitivity could be restored to the mutants by complementation with an intact copy of the sbmA gene. These findings question the viability of the development of insect PR-AMPs as antimicrobials. PMID:24590485

Narayanan, Shalini; Modak, Joyanta K.; Ryan, Catherine S.; Garcia-Bustos, Jose; Davies, John K.

2014-01-01

184

Electrokinetically induced flocculation of Enteroaggregative Escherichia coli  

NASA Astrophysics Data System (ADS)

Enteroaggregative Escherichia coli (EAEC) is a diarrheal microbe, whose aggregative dynamics is involved in its pathogenic behavior. We investigated EAEC's electrokinetic response in miniaturized and microfluidic devices. We found a novel response of the microbe under low magnitude, uniform and oscillating electric fields. In this electrokinetically induced response, microbial adhesion to a glass substrate decreases significantly, leading to a loss of EAEC's biofilm forming abilities. Some earlier studies had indicated that that microbial adhesion and detachment at surfaces can be prompted only by charge-transfer processes at the electrode and not applied electrical potentials - such an inference is not corroborated by our work. Instead, we found that electric fields promote the formation of large mesoscopic microbial aggregations (flocs) in the solution. The presence of frequency dependent relaxation phenomena is explored and the observed results are extended to other microbes.

Kumar, Aloke; Mortensen, Ninell; Harris, Mansueta; Mukherjee, Partha; Retterer, Scott; Doktycz, Mitchel

2011-11-01

185

Animal models of enteroaggregative Escherichia coli infection  

PubMed Central

Enteroaggregative Escherichia coli (EAEC) has been acknowledged as an emerging cause of gastroenteritis worldwide for over two decades. Epidemiologists are revealing the role of EAEC in diarrheal outbreaks as a more common occurrence than ever suggested before. EAEC induced diarrhea is most commonly associated with travelers, children and immunocompromised individuals however its afflictions are not limited to any particular demographic. Many attributes have been discovered and characterized surrounding the capability of EAEC to provoke a potent pro-inflammatory immune response, however cellular and molecular mechanisms underlying initiation, progression and outcomes are largely unknown. This limited understanding can be attributed to heterogeneity in strains and the lack of adequate animal models. This review aims to summarize current knowledge about EAEC etiology, pathogenesis and clinical manifestation. Additionally, current animal models and their limitations will be discussed along with the value of applying systems-wide approaches such as computational modeling to study host-EAEC interactions. PMID:23680797

Philipson, Casandra W.; Bassaganya-Riera, Josep; Hontecillas, Raquel

2013-01-01

186

Genetic Analysis of an Escherichia coli Syndrome  

PubMed Central

A mutant strain of Escherichia coli that fails to recover from prolonged (72 hr) starvation also fails to grow at 43 C. Extracts of this mutant strain show an increased ribonuclease II activity as compared to extracts of the parental strain, and stable ribonucleic acid is degraded to a larger extent in this strain during starvation. Ts+ transductants and revertants were tested for all the above-mentioned phenotypes. All the Ts+ transductants and revertants tested behaved like the Ts+ parental strain, which suggests that all the observed phenotypes are caused by a single sts (starvation-temperature sensitivity) mutation. The reversion rate from sts? to sts+ is rather low but is within the range of reversion rates for other single-site mutations. Three-point transduction crosses located this sts mutation between the ilv and rbs genes. The properties of sts+/sts? merozygotes suggested that the Ts? phenotype of this mutation is recessive. PMID:4945197

Lennette, Evelyne T.; Apirion, David

1971-01-01

187

Direct Upstream Motility in Escherichia coli  

PubMed Central

We provide an experimental demonstration of positive rheotaxis (rapid and continuous upstream motility) in wild-type Escherichia coli freely swimming over a surface. This hydrodynamic phenomenon is dominant below a critical shear rate and robust against Brownian motion and cell tumbling. We deduce that individual bacteria entering a flow system can rapidly migrate upstream (>20 ?m/s) much faster than a gradually advancing biofilm. Given a bacterial population with a distribution of sizes and swim speeds, local shear rate near the surface determines the dominant hydrodynamic mode for motility, i.e., circular or random trajectories for low shear rates, positive rheotaxis for moderate flow, and sideways swimming at higher shear rates. Faster swimmers can move upstream more rapidly and at higher shear rates, as expected. Interestingly, we also find on average that both swim speed and upstream motility are independent of cell aspect ratio. PMID:22500751

Kaya, Tolga; Koser, Hur

2012-01-01

188

Evolution of transcription factors and the gene regulatory network in Escherichia coli  

E-print Network

Evolution of transcription factors and the gene regulatory network in Escherichia coli M. Madan Escherichia coli. In order to gain insight into the evolution of the E.coli regula- tory network, we analysed

Babu, M. Madan

189

Chemotaxis Toward Amino Acids in Escherichia coli  

PubMed Central

Escherichia coli cells are shown to be attracted to the l-amino acids alanine, asparagine, aspartate, cysteine, glutamate, glycine, methionine, serine, and threonine, but not to arginine, cystine, glutamine, histidine, isoleucine, leucine, lysine, phenylalanine, tryptophan, tyrosine, or valine. Bacteria grown in a proline-containing medium were, in addition, attracted to proline. Chemotaxis toward amino acids is shown to be mediated by at least two detection systems, the aspartate and serine chemoreceptors. The aspartate chemoreceptor was nonfunctional in the aspartate taxis mutant, which showed virtually no chemotaxis toward aspartate, glutamate, or methionine, and reduced taxis toward alanine, asparagine, cysteine, glycine, and serine. The serine chemoreceptor was nonfunctional in the serine taxis mutant, which was defective in taxis toward alanine, asparagine, cysteine, glycine, and serine, and which showed no chemotaxis toward threonine. Additional data concerning the specificities of the amino acid chemoreceptors with regard to amino acid analogues are also presented. Finally, two essentially nonoxidizable amino acid analogues, ?-aminoisobutyrate and ?-methylaspartate, are shown to be attractants for E. coli, demonstrating that extensive metabolism of attractants is not required for amino acid taxis. PMID:4562400

Mesibov, Robert; Adler, Julius

1972-01-01

190

Nucleotide excision repair in Escherichia coli.  

PubMed Central

One of the best-studied DNA repair pathways is nucleotide excision repair, a process consisting of DNA damage recognition, incision, excision, repair resynthesis, and DNA ligation. Escherichia coli has served as a model organism for the study of this process. Recently, many of the proteins that mediate E. coli nucleotide excision have been purified to homogeneity; this had led to a molecular description of this repair pathway. One of the key repair enzymes of this pathway is the UvrABC nuclease complex. The individual subunits of this enzyme cooperate in a complex series of partial reactions to bind to and incise the DNA near a damaged nucleotide. The UvrABC complex displays a remarkable substrate diversity. Defining the structural features of DNA lesions that provide the specificity for damage recognition by the UvrABC complex is of great importance, since it represents a unique form of protein-DNA interaction. Using a number of in vitro assays, researchers have been able to elucidate the action mechanism of the UvrABC nuclease complex. Current research is devoted to understanding how these complex events are mediated within the living cell. PMID:2181258

Van Houten, B

1990-01-01

191

Chemotaxis toward sugars in Escherichia coli.  

PubMed

Using a quantitative assay for measuring chemotaxis, we tested a variety of sugars and sugar derivatives for their ability to attract Escherichia coli bacteria. The most effective attractants, i.e., those that have thresholds near 10(-5) M or below, are N-acetyl-d-glucosamine, 6-deoxy-d-glucose, d-fructose, d-fucose, 1-d-glycerol-beta-d-galactoside, galactitol, d-galactose, d-glucosamine, d-glucose, alpha-d-glucose-1-phosphate, lactose, maltose, d-mannitol, d-mannose, methyl-beta-d-galactoside, methyl-beta-d-glucoside, d-ribose, d-sorbitol, and trehalose. Lactose, and probably d-glucose-1-phosphate, are attractive only after conversion to the free monosaccharide, while the other attractants do not require breakdown for taxis. Nine different chemoreceptors are involved in detecting these various attractants. They are called the N-acetyl-glucosamine, fructose, galactose, glucose, maltose, mannitol, ribose, sorbitol, and trehalose chemoreceptors; the specificity of each was studied. The chemoreceptors, with the exception of the one for d-glucose, are inducible. The galactose-binding protein serves as the recognition component of the galactose chemoreceptor. E. coli also has osmotically shockable binding activities for maltose and d-ribose, and these appear to serve as the recognition components for the corresponding chemoreceptors. PMID:4580570

Adler, J; Hazelbauer, G L; Dahl, M M

1973-09-01

192

Independence of replisomes in Escherichia coli chromosomalreplication  

SciTech Connect

In Escherichia coli DNA replication is carried out by the coordinated action of the proteins within a replisome. After replication initiation, the two bidirectionally oriented replisomes from a single origin are colocalized into higher-order structures termed replication factories. The factory model postulated that the two replisomes are also functionally coupled. We tested this hypothesis by using DNA combing and whole-genome microarrays. Nascent DNA surrounding oriC in single, combed chromosomes showed instead that one replisome, usually the leftward one, was significantly ahead of the other 70% of the time. We next used microarrays to follow replication throughout the genome by measuring DNA copy number. We found in multiple E. coli strains that the replisomes are independent, with the leftward replisome ahead of the rightward one. The size of the bias was strain-specific, varying from 50 to 130 kb in the array results. When we artificially blocked one replisome, the other continued unabated, again demonstrating independence. We suggest an improved version of the factory model that retains the advantages of threading DNA through colocalized replisomes at about equal rates, but allows the cell flexibility to overcome obstacles encountered during elongation.

Breier, Adam M.; Weier, Heinz-Ulrich G.; Cozzarelli, Nicholas R.

2005-03-13

193

Molecular characterization and antimicrobial resistance of faecal and urinary Escherichia coli isolated from dogs and humans in Italy.  

PubMed

During this study, 109 faecal Escherichia coli samples isolated from 61 dogs and 48 humans were characterised according to phylogenetic group, extraintestinal virulence factors and antibiotic resistance. The isolates from dogs were predominantly distributed within phylogroup B1 (36%), while the majority of human strains belonged to phylogroup B2 (54%). The prevalence of cnf1, hlyA, papC and sfa virulence genes was significantly associated with the group B2. Canine isolates showed multidrug resistance (MDR) more frequently than human strains. Since group B2 contains most of the strains that cause extraintestinal infections, all 46 B2 faecal strains were confronted against an addition population of 57 urinary E. coli strains belonging to the same phylogroup. The comparison shows that there was no significant difference in the occurrence of virulence factors or in the distribution of antibiotic resistance between faecal and urinary E. coli isolates from dogs. At the same time, a highly significant association was detected between multiple resistance and the source of the strains and between MDR and E. coli isolated from urine in human. This study highlighted similar features of E. coli isolated across sources and hosts. The data suggest a high prevalence of antibiotic resistance in faecal strains, which may represent a serious health risk since these strains can function as a reservoir for uropathogenic E. coli. PMID:24715590

Tramuta, Clara; Robino, Patrizia; Nucera, Daniele; Salvarani, Sara; Banche, Giuliana; Malabaila, Aurelio; Nebbia, Patrizia

2014-01-01

194

Cyclomodulins in Urosepsis Strains of Escherichia coli?  

PubMed Central

Determinants of urosepsis in Escherichia coli remain incompletely defined. Cyclomodulins (CMs) are a growing functional family of toxins that hijack the eukaryotic cell cycle. Four cyclomodulin types are actually known in E. coli: cytotoxic necrotizing factors (CNFs), cycle-inhibiting factor (Cif), cytolethal distending toxins (CDTs), and the pks-encoded toxin. In the present study, the distribution of CM-encoding genes and the functionality of these toxins were investigated in 197 E. coli strains isolated from patients with community-acquired urosepsis (n = 146) and from uninfected subjects (n = 51). This distribution was analyzed in relation to the phylogenetic background, clinical origin, and antibiotic resistance of the strains. It emerged from this study that strains harboring the pks island and the cnf1 gene (i) were strongly associated with the B2 phylogroup (P, <0.001), (ii) frequently harbored both toxin-encoded genes in phylogroup B2 (33%), and (iii) were predictive of a urosepsis origin (P, <0.001 to 0.005). However, the prevalences of the pks island among phylogroup B2 strains, in contrast to those of the cnf1 gene, were not significantly different between fecal and urosepsis groups, suggesting that the pks island is more important for the colonization process and the cnf1 gene for virulence. pks- or cnf1-harboring strains were significantly associated with susceptibility to antibiotics (amoxicillin, cotrimoxazole, and quinolones [P, <0.001 to 0.043]). Otherwise, only 6% and 1% of all strains harbored the cdtB and cif genes, respectively, with no particular distribution by phylogenetic background, antimicrobial susceptibility, or clinical origin. PMID:20375237

Dubois, Damien; Delmas, Julien; Cady, Anne; Robin, Frédéric; Sivignon, Adeline; Oswald, Eric; Bonnet, Richard

2010-01-01

195

Cyclomodulins in urosepsis strains of Escherichia coli.  

PubMed

Determinants of urosepsis in Escherichia coli remain incompletely defined. Cyclomodulins (CMs) are a growing functional family of toxins that hijack the eukaryotic cell cycle. Four cyclomodulin types are actually known in E. coli: cytotoxic necrotizing factors (CNFs), cycle-inhibiting factor (Cif), cytolethal distending toxins (CDTs), and the pks-encoded toxin. In the present study, the distribution of CM-encoding genes and the functionality of these toxins were investigated in 197 E. coli strains isolated from patients with community-acquired urosepsis (n = 146) and from uninfected subjects (n = 51). This distribution was analyzed in relation to the phylogenetic background, clinical origin, and antibiotic resistance of the strains. It emerged from this study that strains harboring the pks island and the cnf1 gene (i) were strongly associated with the B2 phylogroup (P, <0.001), (ii) frequently harbored both toxin-encoded genes in phylogroup B2 (33%), and (iii) were predictive of a urosepsis origin (P, <0.001 to 0.005). However, the prevalences of the pks island among phylogroup B2 strains, in contrast to those of the cnf1 gene, were not significantly different between fecal and urosepsis groups, suggesting that the pks island is more important for the colonization process and the cnf1 gene for virulence. pks- or cnf1-harboring strains were significantly associated with susceptibility to antibiotics (amoxicillin, cotrimoxazole, and quinolones [P, <0.001 to 0.043]). Otherwise, only 6% and 1% of all strains harbored the cdtB and cif genes, respectively, with no particular distribution by phylogenetic background, antimicrobial susceptibility, or clinical origin. PMID:20375237

Dubois, Damien; Delmas, Julien; Cady, Anne; Robin, Frédéric; Sivignon, Adeline; Oswald, Eric; Bonnet, Richard

2010-06-01

196

Phylogenetic grouping and pathotypic comparison of urine and fecal Escherichia coli isolates from children with urinary tract infection.  

PubMed

The aim of this study was to investigate the phylogenetic background and to assess hlyD (involved in the secretion of haemolysin A) and intI1 (encoding a class 1 integrase) in Escherichia coli isolates derived from urinary and fecal specimens. A total of 200 E. coli isolates was collected from patients presenting with urinary tract infection (UTI) during September 2009 to September 2010 and screened for hlyD and intI1 genes by polymerase chain reaction (PCR). Phylogenetic analysis showed that E. coli is composed of four main phylogenetic groups (A, B1, B2 and D) and that uropathogenic E. coli (UPEC) isolates mainly belong to groups B2 (54%) and D (34%) whereas group A (44%) and D (26%) are predominant among commensal E. coli isolates. In this study, hlyD was present in 26% of UPEC and 2% of commensal E. coli isolates. However, hemolytic activity was detected for 42% of UPEC and 6% of commensal E. coli isolates (p < 0.05). intI1 gene was more frequently expressed in UPEC (24%) in comparison with commensal E. coli isolates (12%). Resistance to aztreonam, co-trimoxazole and cefpodoxime were frequently found among UPEC isolates whereas commensal E. coli isolates were commonly resistant to co-trimoxazole, nalidixic acid and cefotaxime. Concluding, a considerable difference between UPEC and commensal E. coli isolates was observed regarding their phylogenetic groups, presence of class 1 integron and hlyD gene, hemolysin activity and resistance pattern. The detection of class 1 integrons and hlyD gene was higher among UPEC compared with commensal E. coli isolates. These findings may contribute for a better understanding of the factors involved in the pathogenesis of UPEC. PMID:25242935

Navidinia, Masoumeh; Peerayeh, Shahin Najar; Fallah, Fatemeh; Bakhshi, Bita; Sajadinia, Raheleh Sadat

2014-01-01

197

COMMUNICATION Escherichia coli tatC Mutations that Suppress Defective  

E-print Network

COMMUNICATION Escherichia coli tatC Mutations that Suppress Defective Twin-Arginine Transporter of the membrane pro- teins TatA, TatB and TatC. Among Gram-positive bacteria, a few organisms can be found

Georgiou, George

198

Structural Sampling of Glycan Interaction Profiles Reveals Mucosal Receptors for Fimbrial Adhesins of Enterotoxigenic Escherichia coli  

PubMed Central

Fimbriae are long, proteinaceous adhesion organelles expressed on the bacterial envelope, evolutionarily adapted by Escherichia coli strains for the colonization of epithelial linings. Using glycan arrays of the Consortium for Functional Glycomics (CFG), the lectin domains were screened of the fimbrial adhesins F17G and FedF from enterotoxigenic E. coli (ETEC) and of the FimH adhesin from uropathogenic E. coli. This has led to the discovery of a more specific receptor for F17G, GlcNAc?1,3Gal. No significant differences emerged from the glycan binding profiles of the F17G lectin domains from five different E. coli strains. However, strain-dependent amino acid variations, predominantly towards the positively charged arginine, were indicated by sulfate binding in FedF and F17G crystal structures. For FedF, no significant binders could be observed on the CFG glycan array. Hence, a shotgun array was generated from microvilli scrapings of the distal jejunum of a 3-week old piglet about to be weaned. On this array, the blood group A type 1 hexasaccharide emerged as a receptor for the FedF lectin domain and remarkably also for F18-fimbriated E. coli. F17G was found to selectively recognize glycan species with a terminal GlcNAc, typifying intestinal mucins. In conclusion, F17G and FedF recognize long glycan sequences that could only be identified using the shotgun approach. Interestingly, ETEC strains display a large capacity to adapt their fimbrial adhesins to ecological niches via charge-driven interactions, congruent with binding to thick mucosal surfaces displaying an acidic gradient along the intestinal tract. PMID:24833052

Lonardi, Emanuela; Moonens, Kristof; Buts, Lieven; de Boer, Arjen R.; Olsson, Johan D. M.; Weiss, Manfred S.; Fabre, Emeline; Guérardel, Yann; Deelder, André M.; Oscarson, Stefan; Wuhrer, Manfred; Bouckaert, Julie

2013-01-01

199

Survival of Escherichia coli in two sewage treatment plants using UV irradiation and chlorination for disinfection.  

PubMed

We investigated the survival of Escherichia coli in two STPs utilising UV irradiation (STP-A) or chlorination (STP-B) for disinfection. In all, 370 E. coli strains isolated from raw influent sewage (IS), secondary treated effluent (STE) and effluent after the disinfection processes of both STPs were typed using a high resolution biochemical fingerprinting method and were grouped into common (C-) and single (S-) biochemical phenotypes (BPTs). In STP-A, 83 BPTs comprising 123 isolates were found in IS and STE, of which 7 BPTs survived UV irradiation. Isolates tested from the same sites of STP-B (n = 220) comprised 122 BPTs, however, only two BPTs were found post-chlorination. A representative isolate from each BPT from both STPs was tested for the presence of 11 virulence genes (VGs) associated with uropathogenic (UPEC) or intestinal pathogenic (IPEC) E. coli strains. Strains surviving UV irradiation were distributed among seven phylogenetic groups with five BPTs carrying VGs associated with either UPEC (4 BPTs) or IPEC (1 BPT). In contrast, E. coli strains found in STP-B carried no VGs. Strains from both STPs were resistant to up to 12 out of the 21 antibiotics tested but there was no significant difference between the numbers of antibiotics to which surviving strains were resistant to in these STPs. Our data suggests that some E. coli strains have a better ability to survive STPs utilising chlorination and UV irradiation for disinfection. However, strains that survive UV irradiation are more diverse and may carry more VGs than those surviving SPTs using chlorination. PMID:24091189

Anastasi, E M; Wohlsen, T D; Stratton, H M; Katouli, M

2013-11-01

200

Structural Sampling of Glycan Interaction Profiles Reveals Mucosal Receptors for Fimbrial Adhesins of Enterotoxigenic Escherichia coli.  

PubMed

Fimbriae are long, proteinaceous adhesion organelles expressed on the bacterial envelope, evolutionarily adapted by Escherichia coli strains for the colonization of epithelial linings. Using glycan arrays of the Consortium for Functional Glycomics (CFG), the lectin domains were screened of the fimbrial adhesins F17G and FedF from enterotoxigenic E. coli (ETEC) and of the FimH adhesin from uropathogenic E. coli. This has led to the discovery of a more specific receptor for F17G, GlcNAcb1,3Gal. No significant differences emerged from the glycan binding profiles of the F17G lectin domains from five different E. coli strains. However, strain-dependent amino acid variations, predominantly towards the positively charged arginine, were indicated by sulfate binding in FedF and F17G crystal structures. For FedF, no significant binders could be observed on the CFG glycan array. Hence, a shotgun array was generated from microvilli scrapings of the distal jejunum of a 3-week old piglet about to be weaned. On this array, the blood group A type 1 hexasaccharide emerged as a receptor for the FedF lectin domain and remarkably also for F18-fimbriated E. coli. F17G was found to selectively recognize glycan species with a terminal GlcNAc, typifying intestinal mucins. In conclusion, F17G and FedF recognize long glycan sequences that could only be identified using the shotgun approach. Interestingly, ETEC strains display a large capacity to adapt their fimbrial adhesins to ecological niches via charge-driven interactions, congruent with binding to thick mucosal surfaces displaying an acidic gradient along the intestinal tract. PMID:24833052

Lonardi, Emanuela; Moonens, Kristof; Buts, Lieven; de Boer, Arjen R; Olsson, Johan D M; Weiss, Manfred S; Fabre, Emeline; Guérardel, Yann; Deelder, André M; Oscarson, Stefan; Wuhrer, Manfred; Bouckaert, Julie

2013-01-01

201

A-type cranberry proanthocyanidins and uropathogenic bacterial anti-adhesion activity  

Microsoft Academic Search

Clinical, epidemiological and mechanistic studies support the role of cranberry (Vaccinium macrocarpon Ait.) in maintaining urinary tract health. Cranberry proanthocyanidins contain A-type linkages and have been associated with preventing adhesion of P-fimbriated uropathogenic Escherichia coli to uroepithelial cells. It is not known if the presence of the A-type linkage is a prerequisite for anti-adhesion activity. Other commercial sources of proanthocyanidins

Amy B. Howell; Jess D. Reed; Christian G. Krueger; Ranee Winterbottom; David G. Cunningham; Marge Leahy

2005-01-01

202

Structure and function of enterotoxigenic Escherichia coli fimbriae from differing assembly pathways.  

PubMed

Pathogenic enterotoxigenic Escherichia coli (ETEC) are the major bacterial cause of diarrhea in young children in developing countries and in travelers, causing significant mortality in children. Adhesive fimbriae are a prime virulence factor for ETEC, initiating colonization of the small intestinal epithelium. Similar to other Gram-negative bacteria, ETEC express one or more diverse fimbriae, some assembled by the chaperone-usher pathway and others by the alternate chaperone pathway. Here, we elucidate structural and biophysical aspects and adaptations of each fimbrial type to its respective host niche. CS20 fimbriae are compared with colonization factor antigen I (CFA/I) fimbriae, which are two ETEC fimbriae assembled via different pathways, and with P-fimbriae from uropathogenic E.?coli. Many fimbriae unwind from their native helical filament to an extended linear conformation under force, thereby sustaining adhesion by reducing load at the point of contact between the bacterium and the target cell. CFA/I fimbriae require the least force to unwind, followed by CS20 fimbriae and then P-fimbriae, which require the highest unwinding force. We conclude from our electron microscopy reconstructions, modeling and force spectroscopy data that the target niche plays a central role in the biophysical properties of fimbriae that are critical for bacterial pathophysiology. PMID:25355550

Mortezaei, Narges; Epler, Chelsea R; Shao, Paul P; Shirdel, Mariam; Singh, Bhupender; McVeigh, Annette; Uhlin, Bernt Eric; Savarino, Stephen J; Andersson, Magnus; Bullitt, Esther

2015-01-01

203

Repair System for Noncanonical Purines in Escherichia coli  

Microsoft Academic Search

Exposure of Escherichia coli strains deficient in molybdopterin biosynthesis (moa) to the purine base N-6-hydroxylaminopurine (HAP) is mutagenic and toxic. We show that moa mutants exposed to HAP also exhibit elevated mutagenesis, a hyperrecombination phenotype, and increased SOS induction. The E. coli rdgB gene encodes a protein homologous to a deoxyribonucleotide triphosphate pyrophosphatase from Methanococ- cus jannaschii that shows a

Nicholas E. Burgis; Jason J. Brucker; Richard P. Cunningham

2003-01-01

204

Escherichia coli Response to Exogenous Pyrophosphate and Analogs  

Microsoft Academic Search

The addition of exogenous pyrophosphate increases the growth yield and cAMP synthesis in stationary phase when Escherichia coli is grown in minimal medium. Pyrophosphate increases the yield by altering the enterobactin uptake system. We studied the physiological effects and examined how the E. coli transcriptome was modified when two structural analogs of pyrophosphate were added to the growth medium. Methylenediphosphonic

Francis Biville; Taku Oshima; Hirotada Mori; Yuya Kawagoe; Odile Bouvet; Marie-Noëlle Rager; Marina Perrotte-Piquemal; Antoine Danchin

2003-01-01

205

ORIGINAL ARTICLE Escherichia coli transcription factor YncC  

E-print Network

, USA Quorum-sensing signal autoinducer 2 (AI-2) stimulates Escherichia coli biofilm formation through is partially reduced by the b3023 mutation. Therefore, AI-2 quorum-sensing control of biofilm formation, which represses biofilm formation of E. coli by repressing motility, inducing the sensor of the quorum-sensing

Wood, Thomas K.

206

Genetic engineering of ethanol production in Escherichia coli  

Microsoft Academic Search

The genes encoding essential enzymes of the fermentative pathway for ethanol production in Zymomonas mobilis, an obligately ethanologenic bacterium, were inserted into Escherichia coli under the control of a common promoter. Alcohol dehydrogenase II and pyruvate decarboxylase from Z. mobilis were expressed at high levels in E. coli, resulting in increased cell growth and the production of ethanol as the

L. O. Ingram; T. Conway; D. P. Clark; G. W. Sewell; J. F. Preston

1987-01-01

207

Molecular Serotyping of Escherichia coli O111:H8  

Technology Transfer Automated Retrieval System (TEKTRAN)

Accurate Escherichia coli serotyping is critical for pathogen diagnosis and surveillance of non-O157 shiga-toxigenic strains, however, few laboratories have this capacity. The molecular serotyping protocol described in this paper targets the somatic and flagellar antigens of E. coli O111:H8 used in...

208

RAPID GLUTAMATE DECARBOXYLASE ASSAY FOR THE DETECTION OF ESCHERICHIA COLI  

EPA Science Inventory

A rapid test procedure for the enzyme glutamate decarboxylase was developed for the detection of Escherichia coli. he assay procedure was able to confirm the presence of E. coli in enteric broth cultures with a 95 percent specificity for both pure cultures and environmental sampl...

209

Multidimensional annotation of the Escherichia coli K-12 genome  

Microsoft Academic Search

The annotation of the Escherichia coli K-12 genome in the EcoCyc database is one of the most accurate, complete and multidimensional genome annota- tions. Of the 4460 E. coli genes, EcoCyc assigns biochemical functions to 76%, and 66% of all genes had their functions determined experimentally. EcoCyc assigns E. coli genes to Gene Ontology and to MultiFun. Seventy-five percent of

Peter D. Karp; Ingrid M. Keseler; Alexander Shearer; Mario Latendresse; Markus Krummenacker; Suzanne M. Paley; Ian Paulsen; Julio Collado-Vides; Socorro Gama-Castro; Martin Peralta-Gil; Alberto Santos-Zavaleta; M. I. Penaloza-Spinola; C. Bonavides-Martinez; J. Ingraham

2007-01-01

210

Biocontrol of Escherichia coli O157  

PubMed Central

The effect of a bacteriophage cocktail (EcoShield™) that is specific against Escherichia coli O157:H7 was evaluated against a nalidixic acid-resistant enterohemorrhagic E. coli O157:H7 RM4407 (EHEC) strain on leafy greens stored under either (1) ambient air or (2) modified atmosphere (MA; 5% O2/35% CO2/60% N2). Pieces (~2 × 2 cm2) of leafy greens (lettuce and spinach) inoculated with 4.5 log CFU/cm2 EHEC were sprayed with EcoShield™ (6.5 log PFU/cm2). Samples were stored at 4 or 10°C for up to 15 d. On spinach, the level of EHEC declined by 2.38 and 2.49 log CFU/cm2 at 4 and 10°C, respectively, 30 min after phage application (p ? 0.05). EcoShield™ was also effective in reducing EHEC on the surface of green leaf lettuce stored at 4°C by 2.49 and 3.28 log units in 30 min and 2 h, respectively (p ? 0.05). At 4°C under atmospheric air, the phage cocktail significantly (p ? 0.05) lowered the EHEC counts in one day by 1.19, 3.21 and 3.25 log CFU/cm2 on spinach, green leaf and romaine lettuce, respectively compared with control (no bacteriophage) treatments. When stored under MA at 4°C, phages reduced (p ? 0.05) EHEC populations by 2.18, 3.50 and 3.13 log CFU/cm2, on spinach, green leaf and romaine lettuce. At 10°C, EHEC reductions under atmospheric air storage were 1.99, 3.90 and 3.99 log CFU/cm2 (p ? 0.05), while population reductions under MA were 3.08, 3.89 and 4.34 logs on spinach, green leaf and romaine lettuce, respectively, compared with controls (p ? 0.05). The results of this study showed that bacteriophages were effective in reducing the levels of E. coli O157:H7 on fresh leafy produce, and that the reduction was further improved when produce was stored under the MA conditions. PMID:23819107

Boyacioglu, Olcay; Sharma, Manan; Sulakvelidze, Alexander; Goktepe, Ipek

2013-01-01

211

Escherichia coli challenge in newborn pigs.  

PubMed

Escherichia coli F18 is a common porcine enteric pathogen causing diarrhea and edema in weaned pigs. An essential step in the pathogenesis of this enteric colibacillosis is a fimbria-receptor interaction in the small intestine, involving the ?(1,2)-fucosyltransferase gene (FUT1) enzyme for bacterial receptor binding to the epithelium. Enzyme expression is genetically determined and increases after weaning at 3 to5 wk, probably due to age- and/or diet-related intestinal maturation. We hypothesized that artificially reared piglets, deprived of sow's milk from birth, show susceptibility to F18 already in the neonatal period. First we verified the intestinal expression of FUT1 in preterm, term, and weaned pigs by quantitative real-time polymerase chain reaction. Then age-related F18 susceptibility (degree of diarrhea) was evaluated in 3-, 10-, and 20-d-old pigs after inoculation of 10(10) cfu E. coli F18 per day for 12 d. Finally, F18 susceptibility was evaluated in caesarean-delivered 0- to 7-d-old piglets inoculated daily with F18 as above. For all pigs, their sows were genotyped to ensure expression of the FUT1 enzyme. FUT1 expression was detected in the proximal and distal small intestine with no apparent differences in levels among preterm, term, and weaned pigs. No consistent F18-induced diarrhea was detected in any of the 3 groups of 3- to 20-d-old pigs. In contrast, 0- to 7-d-old caesarean-delivered pigs showed a higher score of diarrhea in pigs inoculated with F18 compared with controls (2.4 ± 0.1 vs. 1.8 ± 0.1 respectively; P < 0.001). Caesarean-delivered piglets deprived of sow milk are highly susceptible to diarrhea induced by E. coli F18. Lack of the protective effects of birth colonization and sow milk may contribute to high intestinal F18 sensitivity. The newborn pig may be a useful model to investigate factors in maternal milk that protect against F18 diarrhea. PMID:23365279

Jensen, M L; Cilieborg, M S; Østergaard, M V; Bering, S B; Jørgensen, C B; Sangild, P T

2012-12-01

212

Whole-genome phylogeny of Escherichia coli/Shigella group by feature frequency profiles (FFPs)  

PubMed Central

A whole-genome phylogeny of the Escherichia coli/Shigella group was constructed by using the feature frequency profile (FFP) method. This alignment-free approach uses the frequencies of l-mer features of whole genomes to infer phylogenic distances. We present two phylogenies that accentuate different aspects of E. coli/Shigella genomic evolution: (i) one based on the compositions of all possible features of length l = 24 (?8.4 million features), which are likely to reveal the phenetic grouping and relationship among the organisms and (ii) the other based on the compositions of core features with low frequency and low variability (?0.56 million features), which account for ?69% of all commonly shared features among 38 taxa examined and are likely to have genome-wide lineal evolutionary signal. Shigella appears as a single clade when all possible features are used without filtering of noncore features. However, results using core features show that Shigella consists of at least two distantly related subclades, implying that the subclades evolved into a single clade because of a high degree of convergence influenced by mobile genetic elements and niche adaptation. In both FFP trees, the basal group of the E. coli/Shigella phylogeny is the B2 phylogroup, which contains primarily uropathogenic strains, suggesting that the E. coli/Shigella ancestor was likely a facultative or opportunistic pathogen. The extant commensal strains diverged relatively late and appear to be the result of reductive evolution of genomes. We also identify clade distinguishing features and their associated genomic regions within each phylogroup. Such features may provide useful information for understanding evolution of the groups and for quick diagnostic identification of each phylogroup. PMID:21536867

Sims, Gregory E.; Kim, Sung-Hou

2011-01-01

213

Identification of regions of the Escherichia coli chromosome specific for neonatal meningitis-associated strains.  

PubMed

Specific virulence factors associated with the pathogenesis of Escherichia coli strains causing neonatal meningitis (ECNM), such as the K1 capsular polysaccharide, the S fimbriae, and the Ibe10 protein, have been previously identified. However, some other yet unidentified factors are likely to be involved in the pathogenesis of ECNM. To identify specialized unique DNA regions associated with ECNM virulence, we used the representational difference analysis technique. The genomes of two strains belonging to nonpathogenic phylogenetic group A of the ECOR reference collection were subtracted from E. coli strain C5, isolated from a case of neonatal meningitis. Strain C5 belongs to the phylogenetic group B2 as do the majority of ECNM. We have isolated and mapped 64 DNA fragments which are specific for strain C5 and not found in nonpathogenic strains. Of these clones, 44 were clustered in six distinct regions on the chromosome. The sfa and ibe10 genes were located in regions 2 and 6, respectively. A group of genes (cnf1, hra, hly, and prs) known to be present in a pathogenicity island of the uropathogenic strain E. coli J96 colocalized with region 6. The occurrence of these DNA regions was tested in a set of meningitis-associated strains and in a control group composed of non-meningitis-associated strains belonging to the same B2 group. Regions 1, 3, and 4 were present in 91, 82, and 81%, respectively, of the meningitis strains and in 40, 13, and 47% of the control strains. Together, these data suggest that regions 1, 3, and 4 code for factors associated with the ability of E. coli to invade the meninges of neonates. PMID:10722606

Bonacorsi, S P; Clermont, O; Tinsley, C; Le Gall, I; Beaudoin, J C; Elion, J; Nassif, X; Bingen, E

2000-04-01

214

Type 1 Fimbriae Contribute to Catheter-Associated Urinary Tract Infections Caused by Escherichia coli  

PubMed Central

Biofilm formation on catheters is thought to contribute to persistence of catheter-associated urinary tract infections (CAUTI), which represent the most frequent nosocomial infections. Knowledge of genetic factors for catheter colonization is limited, since their role has not been assessed using physicochemical conditions prevailing in a catheterized human bladder. The current study aimed to combine data from a dynamic catheterized bladder model in vitro with in vivo expression analysis for understanding molecular factors relevant for CAUTI caused by Escherichia coli. By application of the in vitro model that mirrors the physicochemical environment during human infection, we found that an E. coli K-12 mutant defective in type 1 fimbriae, but not isogenic mutants lacking flagella or antigen 43, was outcompeted by the wild-type strain during prolonged catheter colonization. The importance of type 1 fimbriae for catheter colonization was verified using a fimA mutant of uropathogenic E. coli strain CFT073 with human and artificial urine. Orientation of the invertible element (IE) controlling type 1 fimbrial expression in bacterial populations harvested from the colonized catheterized bladder in vitro suggested that the vast majority of catheter-colonizing cells (up to 88%) express type 1 fimbriae. Analysis of IE orientation in E. coli populations harvested from patient catheters revealed that a median level of ?73% of cells from nine samples have switched on type 1 fimbrial expression. This study supports the utility of the dynamic catheterized bladder model for analyzing catheter colonization factors and highlights a role for type 1 fimbriae during CAUTI. PMID:24336940

Maierl, Mario; Jörger, Michael; Krause, Robert; Berger, Daniela; Haid, Andrea; Tesic, Dijana; Zechner, Ellen L.

2014-01-01

215

Regulation of alcohol fermentation by Escherichia coli  

SciTech Connect

The purpose of this project is to elucidate the way in which the fermentative synthesis of ethanol is regulated in the facultative anaerobe Escherichia coli. Focus is on the two final steps in alcohol synthesis, which are catalyzed by alcohol dehydrogenase and acetaldehyde CoA dehydrogenase. We have isolated a series of mutations affecting the expression of these enzymes. Some of these mutations are in the structural genes for these enzymes; others affect the regulation of the adh operon. We have recently cloned the genes coding for these enzymes and are now studying the effect of multiple copies of the adh gene on fermentative growth and its regulation. A recently invented technique, proton suicide has allowed the selection of a variety of novel mutants affecting fermentation which are presently being characterized. We have isolated a comprehensive collection of operon fusions in which the lacZ structural gene is fused to promoters that are inactive aerobically but active anaerobically. Although these genes (like adh) are only expressed under anaerobic conditions, the level of induction varies from two-fold to nearly 100-fold. The nitrogen source, medium pH, nature of the buffer, presence of alternative electron acceptors (e.g., nitrate), and other factors exert a great effect on the expression of many of these genes. In the near future we will investigate control mechanisms common to the adh operon and other anaerobically regulated genes.

Clark, D.P.

1986-03-01

216

Biochemistry of homologous recombination in Escherichia coli.  

PubMed Central

Homologous recombination is a fundamental biological process. Biochemical understanding of this process is most advanced for Escherichia coli. At least 25 gene products are involved in promoting genetic exchange. At present, this includes the RecA, RecBCD (exonuclease V), RecE (exonuclease VIII), RecF, RecG, RecJ, RecN, RecOR, RecQ, RecT, RuvAB, RuvC, SbcCD, and SSB proteins, as well as DNA polymerase I, DNA gyrase, DNA topoisomerase I, DNA ligase, and DNA helicases. The activities displayed by these enzymes include homologous DNA pairing and strand exchange, helicase, branch migration, Holliday junction binding and cleavage, nuclease, ATPase, topoisomerase, DNA binding, ATP binding, polymerase, and ligase, and, collectively, they define biochemical events that are essential for efficient recombination. In addition to these needed proteins, a cis-acting recombination hot spot known as Chi (chi: 5'-GCTGGTGG-3') plays a crucial regulatory function. The biochemical steps that comprise homologous recombination can be formally divided into four parts: (i) processing of DNA molecules into suitable recombination substrates, (ii) homologous pairing of the DNA partners and the exchange of DNA strands, (iii) extension of the nascent DNA heteroduplex; and (iv) resolution of the resulting crossover structure. This review focuses on the biochemical mechanisms underlying these steps, with particular emphases on the activities of the proteins involved and on the integration of these activities into likely biochemical pathways for recombination. Images PMID:7968921

Kowalczykowski, S C; Dixon, D A; Eggleston, A K; Lauder, S D; Rehrauer, W M

1994-01-01

217

Membrane topology of Escherichia coli diacylglycerol kinase.  

PubMed Central

The topology of Escherichia coli diacylglycerol kinase (DAGK) within the cytoplasmic membrane was elucidated by a combined approach involving both multiple aligned sequence analysis and fusion protein experiments. Hydropathy plots of the five prokaryotic DAGK sequences available were uniform in their prediction of three transmembrane segments. The hydropathy predictions were experimentally tested genetically by fusing C-terminal deletion derivatives of DAGK to beta-lactamase and beta-galactosidase. Following expression, the enzymatic activities of the chimeric proteins were measured and used to determine the cellular location of the fusion junction. These studies confirmed the hydropathy predictions for DAGK with respect to the number and approximate sequence locations of the transmembrane segments. Further analysis of the aligned DAGK sequences detected probable alpha-helical N-terminal capping motifs and two amphipathic alpha-helices within the enzyme. The combined fusion and sequence data indicate that DAGK is a polytopic integral membrane protein with three transmembrane segments with the N terminus of the protein in the cytoplasm, the C terminus in the periplasmic space, and two amphipathic helices near the cytoplasmic surface. Images PMID:8071224

Smith, R L; O'Toole, J F; Maguire, M E; Sanders, C R

1994-01-01

218

Antimicrobial-resistant Invasive Escherichia coli, Spain  

PubMed Central

To address the public health problem of antimicrobial resistance, the European Union founded the European Antimicrobial Resistance Surveillance System. A network of 32 Spanish hospitals, serving ?9.6 million persons, submitted antimicrobial-susceptibility data on 7,098 invasive Escherichia coli species (2001–2003). Resistance to ampicillin, cotrimoxazole, ciprofloxacin, gentamicin, and tobramycin was found at rates of 59.9%, 32.6%, 19.3%, 6.8%, and 5.3%, respectively. Resistance to multiple drugs increased from 13.8% in 2001 to 20.6% in 2003 (p <0.0001). Antimicrobial consumption data were obtained from the Spanish National Health System. In spite of decreased cephalosporin and ?-lactam use, overall extended-spectrum ?-lactamase production increased from 1.6% (2001) to 4.1% (2003) (p <0.0001), mainly due to the rising prevalence of cefotaximases. Resistance to ciprofloxacin significantly increased, mostly in community-onset infections, which coincided with a rise in community quinolone use. Cotrimoxazole resistance remained stable at ?30%, even though its use was dramatically reduced. PMID:15829192

Oteo, Jesús; Lázaro, Edurne; de Abajo, Francisco J.; Baquero, Fernando; Campos, José

2005-01-01

219

Escherichia coli mutants impaired in maltodextrin transport.  

PubMed Central

Wild-type Escherichia coli K-12 was found to grow equally well on maltose and on maltodextrins containing up to seven glucose residues. Three classes of mutants unable to grow on maltodextrins, but still able to grow on maltose, were investigated in detail. The first class, already known, was composed of phage lambda-resistant mutants, which lack the outer membrane protein coded by gene lamB. These mutants grow on maltose and maltotriose but not at all on maltotetraose and longer maltodextrins which cannot cross the outer membrane. A second class of mutants were affected in malE, the structural gene of the periplasmic maltose binding protein. The maltose binding proteins isolated from the new mutants were altered in their substrate binding properties, but not in a way that could account for the mutant phenotypes. Rather, the results of growth experiments and transport studies suggest that these malE mutants are impaired in their ability to transport maltodextrins across the outer membrane. This implies that the maltose binding protein (in wild-type strains) cooperates with the lambda receptor in permeation through the outer membrane. The last class of mutants described in this paper were affected in malG, or perhaps in an as yet undetected gene close to malG. They were defective in the transfer of maltodextrins from the periplasmic space to the cytoplasm but only slightly affected in the transport of maltose. Images PMID:387714

Wandersman, C; Schwartz, M; Ferenci, T

1979-01-01

220

Incomplete flagellar structures in Escherichia coli mutants.  

PubMed Central

Escherichia coli mutants with defects in 29 flagellar genes identified so far were examined by electron microscopy for possession of incomplete flagellar structures in membrane-associated fractions. The results are discussed in consideration of the known transcriptional interaction of flagellar genes. Hook-basal body structures were detected in flaD, flaS, flaT, flbC, and hag mutants. The flaE mutant had a polyhook-basal body structure. An intact basal body appeared in flaK mutants. Putative precursors of the basal body were detected in mutants with defects in flaM, flaU, flaV, and flaY. No structures homologous to the flagellar basal body or its parts were detected in mutants with defects in flaA, flaB, flaC, flaG, flaH, flaI, flaL, flaN, flaO, flaP, flaQ, flaR, flaW, flaX, flbA, flbB, and flbD. One flaZ mutant had an incomplete flagellar basal body structure and another formed no significant structure, suggesting that flaZ is responsible for both basal body assembly and the transcription of the hag gene. Images PMID:7007337

Suzuki, T; Komeda, Y

1981-01-01

221

Virulence regulons of enterotoxigenic Escherichia coli.  

PubMed

Enterotoxigenic Escherichia coli is frequently associated with travelers' diarrhea and is a leading cause of infant and childhood mortality in developing countries. Disease is dependent upon the orchestrated expression of enterotoxins, flexible adhesive pili, and other virulence factors. Both the heat-labile (LT) and heat-stable (ST-H) enterotoxins are regulated at the level of transcription by cAMP-receptor protein which represses the expression of LT while activating expression of ST-H. The expression of many different serotypes of adhesive pili is regulated by Rns, a member of the AraC family that represents a subgroup of conserved virulence regulators from several enteric pathogens. These Rns-like regulators recognize similar DNA binding sites, and a compiled sequence logo suggests they may bind DNA through both major and minor groove interactions. These regulators are also tempting targets for novel therapeutics because they play pivotal roles during infection. To that end, high-throughput screens have begun to identify compounds that inhibit the activity of these regulators, predominately by interfering with DNA binding. PMID:24203442

Munson, George P

2013-12-01

222

The Escherichia coli divisome: born to divide.  

PubMed

Septation in Escherichia coli involves complex molecular mechanisms that contribute to the accuracy of bacterial division. The proto-ring, a complex made up by the FtsZ, FtsA and ZipA proteins, forms at the beginning of the process and directs the assembly of the full divisome. Central to this complex is the FtsZ protein, a GTPase able to assemble into a ring-like structure that responds to several modulatory inputs including mechanisms to position the septum at midcell. The connection with the cell wall synthesising machinery stabilizes the constriction of the cytoplasmic membrane. Although a substantial amount of evidence supports this description, many details on how individual divisome elements are structured or how they function are subjected to controversial interpretations. We discuss these discrepancies arising from incomplete data and from technical difficulties imposed by the small size of bacteria. Future work, including more powerful imaging and reconstruction technologies, will help to clarify the missing details on the architecture and function of the bacterial division machinery. PMID:23962168

Natale, Paolo; Pazos, Manuel; Vicente, Miguel

2013-12-01

223

Tamm-Horsfall protein binds to type 1 fimbriated Escherichia coli and prevents E. coli from binding to uroplakin Ia and Ib receptors.  

PubMed

The adherence of uropathogenic Escherichia coli to the urothelial surface, a critical first step in the pathogenesis of urinary tract infection (UTI), is controlled by three key elements: E. coli adhesins, host receptors, and host defense mechanisms. Although much has been learned about E. coli adhesins and their urothelial receptors, little is known about the role of host defense in the adherence process. Here we show that Tamm-Horsfall protein (THP) is the principal urinary protein that binds specifically to type 1 fimbriated E. coli, the main cause of UTI. The binding was highly specific and saturable and could be inhibited by d-mannose and abolished by endoglycosidase H treatment of THP, suggesting that the binding is mediated by the high-mannose moieties of THP. It is species-conserved, occurring in both human and mouse THPs. In addition, the binding to THP was much greater with an E. coli strain bearing a phenotypic variant of the type 1 fimbrial FimH adhesin characteristic of those prevalent in UTI isolates compared with the one prevalent in isolates from the large intestine of healthy individuals. Finally, a physiological concentration of THP completely abolished the binding of type 1 fimbriated E. coli to uroplakins Ia and Ib, two putative urothelial receptors for type 1 fimbriae. These results establish, on a functional level, that THP contains conserved high-mannose moieties capable of specific interaction with type 1 fimbriae and strongly suggest that this major urinary glycoprotein is a key urinary anti-adherence factor serving to prevent type 1 fimbriated E. coli from binding to the urothelial receptors. PMID:11134021

Pak, J; Pu, Y; Zhang, Z T; Hasty, D L; Wu, X R

2001-03-30

224

Shiga toxin-producing Escherichia coli.  

PubMed

In the United States, it is estimated that non-O157 Shiga toxin-producing Escherichia coli (STEC) cause more illnesses than STEC O157:H7, and the majority of cases of non-O157 STEC infections are due to serogroups O26, O45, O103, O111, O121, and O145, referred to as the top six non-O157 STEC. The diseases caused by non-O157 STEC are generally milder than those induced by O157 STEC; nonetheless, non-O157 STEC strains have also been associated with serious illnesses such as hemorrhagic colitis and hemolytic uremic syndrome, as well as death. Ruminants, particularly cattle, are reservoirs for both O157 and non-O157 STEC, which are transmitted to humans by person-to-person or animal contact and by ingestion of food or water contaminated with animal feces. Improved strategies to control STEC colonization and shedding in cattle and contamination of meat and produce are needed. In general, non-O157 STEC respond to stresses such as acid, heat, and other stresses induced during food preparation similar to O157 STEC. Similar to O157:H7, the top six non-O157 STEC are classified as adulterants in beef by the USDA Food Safety and Inspection Service, and regulatory testing for these pathogens began in June 2012. Due to the genetic and phenotypic variability of non-O157 STEC strains, the development of accurate and reliable methods for detection and isolation of these pathogens has been challenging. Since the non-O157 STEC are responsible for a large portion of STEC-related illnesses, more extensive studies on their physiology, genetics, pathogenicity, and evolution are needed in order to develop more effective control strategies. PMID:24377855

Smith, James L; Fratamico, Pina M; Gunther, Nereus W

2014-01-01

225

Severe Zinc Depletion of Escherichia coli  

PubMed Central

Zinc ions play indispensable roles in biological chemistry. However, bacteria have an impressive ability to acquire Zn2+ from the environment, making it exceptionally difficult to achieve Zn2+ deficiency, and so a comprehensive understanding of the importance of Zn2+ has not been attained. Reduction of the Zn2+ content of Escherichia coli growth medium to 60 nm or less is reported here for the first time, without recourse to chelators of poor specificity. Cells grown in Zn2+-deficient medium had a reduced growth rate and contained up to five times less cellular Zn2+. To understand global responses to Zn2+ deficiency, microarray analysis was conducted of cells grown under Zn2+-replete and Zn2+-depleted conditions in chemostat cultures. Nine genes were up-regulated more than 2-fold (p < 0.05) in cells from Zn2+-deficient chemostats, including zinT (yodA). zinT is shown to be regulated by Zur (zinc uptake regulator). A mutant lacking zinT displayed a growth defect and a 3-fold lowered cellular Zn2+ level under Zn2+ limitation. The purified ZinT protein possessed a single, high affinity metal-binding site that can accommodate Zn2+ or Cd2+. A further up-regulated gene, ykgM, is believed to encode a non-Zn2+ finger-containing paralogue of the Zn2+ finger ribosomal protein L31. The gene encoding the periplasmic Zn2+-binding protein znuA showed increased expression. During both batch and chemostat growth, cells “found” more Zn2+ than was originally added to the culture, presumably because of leaching from the culture vessel. Zn2+ elimination is shown to be a more precise method of depleting Zn2+ than by using the chelator N,N,N?,N?-tetrakis(2-pyridylmethyl)ethylenediamine. PMID:19377097

Graham, Alison I.; Hunt, Stuart; Stokes, Sarah L.; Bramall, Neil; Bunch, Josephine; Cox, Alan G.; McLeod, Cameron W.; Poole, Robert K.

2009-01-01

226

Inhibition of Escherichia coli CFT073 fliC Expression and Motility by Cranberry Materials ?  

PubMed Central

In humans, uropathogenic Escherichia coli (UPEC) is the most common etiological agent of uncomplicated urinary tract infections (UTIs). Cranberry extracts have been linked to the prevention of UTIs for over a century; however, a mechanistic understanding of the way in which cranberry derivatives prevent bacterial infection is still lacking. In this study, we used a fliC-lux reporter as well as quantitative reverse transcription-PCR to demonstrate that when UPEC strain CFT073 was grown or exposed to dehydrated, crushed cranberries or to purified cranberry-derived proanthocyanidins (cPACs), expression of the flagellin gene (fliC) was inhibited. In agreement with these results, transmission electron microscopy imaging of bacteria grown in the presence of cranberry materials revealed fewer flagella than those in bacteria grown under control conditions. Furthermore, we showed that swimming and swarming motilities were hindered when bacteria were grown in the presence of the cranberry compounds. Because flagellum-mediated motility has been suggested to enable UPEC to disseminate to the upper urinary tract, we propose that inhibition of flagellum-mediated motility might be a key mechanism by which cPACs prevent UTIs. This is the first report to show that cranberry compounds inhibit UPEC motility via downregulation of the fliC gene. Further studies are required to establish whether these inhibitors play a role in vivo. PMID:21821749

Hidalgo, Gabriela; Chan, Michelle; Tufenkji, Nathalie

2011-01-01

227

Comparison of the PhoPQ Regulon in Escherichia coli and Salmonella typhimurium  

E-print Network

Comparison of the PhoPQ Regulon in Escherichia coli and Salmonella typhimurium Pieter Monsieurs,1 as a transcriptional regulator that responds to Mg2+ starvation both in Escherichia coli and Salmonella typhimurium.g., pathogenesis in S. typhimurium). Key words: PhoPQ regulon -- Escherichia coli -- Salmonella typhimirium

228

Obscured phylogeny and possible recombinational dormancy in Escherichia coli  

PubMed Central

Background Escherichia coli is one of the best studied organisms in all of biology, but its phylogenetic structure has been difficult to resolve with current data and analytical techniques. We analyzed single nucleotide polymorphisms in chromosomes of representative strains to reconstruct the topology of its emergence. Results The phylogeny of E. coli varies according to the segment of chromosome analyzed. Recombination between extant E. coli groups is largely limited to only three intergroup pairings. Conclusions Segment-dependent phylogenies most likely are legacies of a complex recombination history. However, E. coli are now in an epoch in which they no longer broadly share DNA. Using the definition of species as organisms that freely exchange genetic material, this recombinational dormancy could reflect either the end of E. coli as a species, or herald the coalescence of E. coli groups into new species. PMID:21708031

2011-01-01

229

Pathogenomics of the Virulence Plasmids of Escherichia coli  

PubMed Central

Summary: Bacterial plasmids are self-replicating, extrachromosomal elements that are key agents of change in microbial populations. They promote the dissemination of a variety of traits, including virulence, enhanced fitness, resistance to antimicrobial agents, and metabolism of rare substances. Escherichia coli, perhaps the most studied of microorganisms, has been found to possess a variety of plasmid types. Included among these are plasmids associated with virulence. Several types of E. coli virulence plasmids exist, including those essential for the virulence of enterotoxigenic E. coli, enteroinvasive E. coli, enteropathogenic E. coli, enterohemorrhagic E. coli, enteroaggregative E. coli, and extraintestinal pathogenic E. coli. Despite their diversity, these plasmids belong to a few plasmid backbones that present themselves in a conserved and syntenic manner. Thanks to some recent research, including sequence analysis of several representative plasmid genomes and molecular pathogenesis studies, the evolution of these virulence plasmids and the implications of their acquisition by E. coli are now better understood and appreciated. Here, work involving each of the E. coli virulence plasmid types is summarized, with the available plasmid genomic sequences for several E. coli pathotypes being compared in an effort to understand the evolution of these plasmid types and define their core and accessory components. PMID:19946140

Johnson, Timothy J.; Nolan, Lisa K.

2009-01-01

230

Recombinational Construction in Escherichia coli of Infectious Adenoviral Genomes  

Microsoft Academic Search

A two-step gene replacement procedure was developed that generates infectious adenoviral genomes through homologous recombination in Escherichia coli. As a prerequisite, a human adenovirus serotype 5 (Ad5)-derived genome was first introduced as a PacI restriction fragment into an incP-derived replicon which, in contrast to ColE1-derivatives (e.g., pBR322 or pUC plasmids), is functional in a polA mutant of E. coli. Any

Joel Crouzet; Laurent Naudin; Cecile Orsini; Emmanuelle Vigne; Lucy Ferrero; Aude Le Roux; Patrick Benoit; Martine Latta; Christophe Torrent; Didier Branellec; Patrice Denefle; Jean-Francois Mayaux; Michel Perricaudet; Patrice Yeh

1997-01-01

231

EcoCyc: a comprehensive database resource for Escherichia coli  

Microsoft Academic Search

The EcoCyc database (http:\\/\\/EcoCyc.org\\/) is a com- prehensivesourceofinformationonthebiologyofthe prototypical model organism Escherichia coli K12. ThemissionforEcoCycistocontainbothcomputable descriptions of, and detailed comments describing, all genes, proteins, pathways and molecular interac- tions in E.coli. Through ongoing manual curation, extensive information such as summary comments, regulatory information, literature citations and evi- dence types has been extracted from 8862 publica- tions and added to Version

Ingrid M. Keseler; Julio Collado-vides; Socorro Gama-castro; John Ingraham; Suzanne M. Paley; Ian T. Paulsen; Martín Peralta-gil; Peter D. Karp

2005-01-01

232

EcoCyc: a comprehensive database resource for Escherichia coli  

Microsoft Academic Search

The EcoCyc database (http:\\/\\/EcoCyc. org\\/) is a com-prehensive source of information on the biology of the prototypical model organism Escherichia coli K12.The mission for EcoCyc is to contain both computable descriptions of, and detailed comments describing, all genes, proteins, pathways and molecular interac-tions in E. coli. Through ongoing manual curation, extensive information such as summary comments, regulatory information, literature citations

Ingrid M. Keseler; Julio Collado-vides; Socorro Gama-castro; John Ingraham

2006-01-01

233

YeeO from Escherichia coli exports flavins.  

PubMed

Multidrug and toxic compound extrusion (MATE) proteins help maintain cellular homeostasis by secreting metabolic wastes. Flavins may occur as cellular waste products, with their production and secretion providing potential benefit for industrial applications related to biofuel cells. Here we find that MATE protein YeeO from Escherichia coli exports both flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD). Significant amounts of flavins were trapped intracellularly when YeeO was produced indicating transport limits secretion of flavins. Wild-type E. coli secreted 3 flavins (riboflavin, FMN, and FAD), so E. coli likely produces additional flavin transporters. PMID:25482085

McAnulty, Michael J; Wood, Thomas K

2014-11-01

234

Sources of Escherichia coli in a coastal subtropical environment.  

PubMed

Sources of Escherichia coli in a coastal waterway located in Ft. Lauderdale, Fla., were evaluated. The study consisted of an extensive program of field measurements designed to capture spatial and temporal variations in E. coli concentrations as well as experiments conducted under laboratory-controlled conditions. E. coli from environmental samples was enumerated by using a defined substrate technology (Colilert-18). Field sampling tasks included sampling the length of the North Fork to identify the river reach contributing high E. coli levels, autosampler experiments at two locations, and spatially intense sampling efforts at hot spots. Laboratory experiments were designed to simulate tidal conditions within the riverbank soils. The results showed that E. coli entered the river in a large pulse during storm conditions. After the storm, E. coli levels returned to baseline levels and varied in a cyclical pattern which correlated with tidal cycles. The highest concentrations were observed during high tide, whereas the lowest were observed at low tide. This peculiar pattern of E. coli concentrations between storm events was caused by the growth of E. coli within riverbank soils which were subsequently washed in during high tide. Laboratory analysis of soil collected from the riverbanks showed increases of several orders of magnitude in soil E. coli concentrations. The ability of E. coli to multiply in the soil was found to be a function of soil moisture content, presumably due to the ability of E. coli to outcompete predators in relatively dry soil. The importance of soil moisture in regulating the multiplication of E. coli was found to be critical in tidally influenced areas due to periodic wetting and drying of soils in contact with water bodies. Given the potential for growth in such systems, E. coli concentrations can be artificially elevated above that expected from fecal impacts alone. Such results challenge the use of E. coli as a suitable indicator of water quality in tidally influenced areas located within tropical and subtropical environments. PMID:10618229

Solo-Gabriele, H M; Wolfert, M A; Desmarais, T R; Palmer, C J

2000-01-01

235

The presence of genes encoding for different virulence factors in clonally related Escherichia coli that produce CTX-Ms.  

PubMed

Successful international clones have recently emerged among Escherichia coli that produce CTX-M ?-lactamases as important causes of community-onset urinary tract and bloodstream infections. One hundred and seven isolates that belong to sequence types (STs) ST38, ST131, ST405, ST648, and 38 nonrelated CTX-M-producing E. coli from Canada and the Netherlands were assigned to phylogenetic groups and tested for the presence of genes encoding for virulence factors (VFs) using established multiplex polymerase chain reaction. The STs E. coli were significantly more resistant to antibiotics--ST38, ST405, and ST648 belonged to phylogenetic group D while ST131 belonged to B2. Secreted autotransporter toxin (sat), aerobactin receptor, and pathogenicity island marker were significantly more common among the STs; the heat-resistant agglutinin (hra) was present in ST38, sat, and uropathogenic-specific protein, and putative adhesin-siderophore receptor was more common in ST131, while outer membrane protease T was present in ST648. ST131 had a significantly higher VF score. In conclusion, the precise role of these VFs remains to be elucidated; however, we have identified certain putative VFs that possibly contribute to the fitness and success of certain sequence types. PMID:22300954

Van der Bij, Akke K; Peirano, Gisele; Pitondo-Silva, André; Pitout, Johann D D

2012-04-01

236

Slugs: Potential Novel Vectors of Escherichia coli O157  

PubMed Central

Field and laboratory studies were performed to determine whether slugs could act as novel vectors for pathogen (e.g., Escherichia coli O157) transfer from animal feces to salad vegetables. Escherichia coli O157 was isolated from 0.21% of field slugs from an Aberdeenshire sheep farm. These isolates carried the verocytotoxin genes (vt1 and vt2) and the attaching and effacing gene (eae), suggesting that they are potentially pathogenic to humans. Strain typing using multilocus variable number tandem repeats analysis showed that slug and sheep isolates were indistinguishable. Laboratory experiments using an E. coli mutant resistant to nalidixic acid showed that the ubiquitous slug species Deroceras reticulatum could carry viable E. coli on its external surface for up to 14 days. Slugs that had been fed E. coli shed viable bacteria in their feces with numbers showing a short but statistically significant linear log decline. Further, it was found that E. coli persisted for up to 3 weeks in excreted slug feces, and hence, we conclude that slugs have the potential to act as novel vectors of E. coli O157. PMID:16391036

Sproston, Emma L.; Macrae, M.; Ogden, Iain D.; Wilson, Michael J.; Strachan, Norval J. C.

2006-01-01

237

Naturalized Escherichia coli from New Zealand wetland and stream environments.  

PubMed

This research investigates the presence of a naturalized clade of Escherichia coli in wetland and stream biofilms. Escherichia coli is used as a faecal indicator in water quality monitoring programmes worldwide, with the assumption that this bacterium is exclusively a commensal of the vertebrate gut. However, recent findings indicate growth and multiplication of E. coli in water and soils. This study seeks to clarify the relationships between environmental and commensal E. coli strains retrieved from New Zealand streams by evaluating fundamental genetic differences using the multilocus sequence typing (MLST) method. Environmental and commensal strains showed a high diversity of MLST profiles. Genetic analyses of linkage disequilibrium, index of association and rates of synonymous and nonsynonymous substitutions were used to investigate sequence variability and nature of change. Phylogenetic trees based on the concatenated sequences of the seven MLST housekeeping genes displayed distinct clustering of environmental strains. Comparison of the New Zealand sequences with worldwide E. coli strains retrieved from the Shigatox MLST database online did not allow the identification of a clear environmental genotype. However, some New Zealand aquatic E. coli isolates showed close relationships with strains from human and bovine origins, suggesting that environmental isolates were originally derived from subpopulations of commensal E. coli from these sources. PMID:22974403

Perchec-Merien, Anne-Marie; Lewis, Gillian D

2013-02-01

238

The hydrogenases and formate dehydrogenases of Escherichia coli  

Microsoft Academic Search

Escherichia coli has the capacity to synthesise three distinct formate dehydrogenase isoenzymes and three hydrogenase isoenzymes. All six are multisubunit, membrane-associated proteins that are functional in the anaerobic metabolism of the organism. One of the formate dehydrogenase isoenzymes is also synthesised in aerobic cells. Two of the formate dehydrogenase enzymes and two hydrogenases have a respiratory function while the formate

Gary Sawers

1994-01-01

239

Synthesis of Calf Prochymosin (Prorennin) in Escherichia coli  

Microsoft Academic Search

A gene for calf prochymosin (prorennin) has been reconstructed from chemically synthesized oligodeoxyribonucleotides and cloned DNA copies of preprochymosin mRNA. This gene has been inserted into a bacterial expression plasmid containing the Escherichia coli tryptophan promoter and a bacterial ribosome binding site. Induction of transcription from the tryptophan promoter results in prochymosin synthesis at a level of up to 5%

J. S. Emtage; S. Angal; M. T. Doel; T. J. R. Harris; B. Jenkins; G. Lilley; P. A. Lowe

1983-01-01

240

Escherichia coli bacteraemia in Canberra: incidence and clinical features  

Microsoft Academic Search

Objective: To determine the population incidence and clinical features of Escherichia coli bacteraemia in Canberra, Australia. Design, setting and participants: Canberra (including the nearby local government areas of Queanbeyan and Yarrowlumla) has a geographically isolated population of about 366 000 people. Its six hospitals also provide tertiary medical services for the surrounding region. Confining our analysis (by residential postcodes) to

Karina J Kennedy; Jan L Roberts; Peter J Collignon

2008-01-01

241

Combating enteropathogenic Escherichia coli (EPEC) infections: the way forward  

PubMed Central

Enteropathogenic Escherichia coli (EPEC) strains continue to cause severe and sometimes fatal infantile diarrhea, particularly in Africa. Increased efforts at diagnosis, defining the clinical spectrum of disease, understanding pathogenic mechanisms, and delineating immune responses are desperately needed to develop new strategies to combat EPEC. PMID:23815982

Donnenberg, Michael S.; Finlay, B. Brett

2015-01-01

242

Why the Phosphotransferase System of Escherichia coli Escapes Diffusion Limitation  

Microsoft Academic Search

We calculated the implications of diffusion for the phosphoenolpyruvate:glucose phosphotransferase system (glucose-PTS) of Escherichia coli in silicon cells of various magnitudes. For a cell of bacterial size, diffusion limitation of glucose influx was negligible. Nevertheless, a significant concentration gradient for one of the enzyme species, nonphosphorylated IIAGlc, was found. This should have consequences because the phosphorylation state of IIAGlc is

Christof Francke; Pieter W. Postma; Hans V. Westerhoff; Joke G. Blom; Mark A. Peletier

2003-01-01

243

Escherichia coli and other Enterobacteriaceae: Food poisoning and health effects  

Technology Transfer Automated Retrieval System (TEKTRAN)

The family Enterobactericeae consists of rod-shaped, Gram-negative, facultatively anaerobic, non-spore forming bacteria and also includes the food-borne pathogens, Cronobacter spp., Escherichia coli, Salmonella enterica, Shigella spp., and Yersinia spp. Illness caused by these pathogens is acquired...

244

Inactivation of Escherichia coli by titanium dioxide photocatalytic oxidation.  

EPA Science Inventory

Titanium dioxide in the anatase crystalline form was used as a photocatalyst to generate hydroxyl radicals in a flowthrough water reactor. Experiments were performed on pure cultures of Escherichia coli in dechlorinated tap water and a surface water sample to evaluate the disinfe...

245

Global Incidence of Carbapenemase-Producing Escherichia coli ST131  

PubMed Central

We characterized Escherichia coli ST131 isolates among 116 carbapenemase-producing strains. Of isolates from 16 countries collected during 2008–2013, 35% belonged to ST131 and were associated with blaKPC, H30 lineage, and virotype C. This study documents worldwide incidents of resistance to “last resort” antimicrobial drugs among a common pathogen in a successful sequence type. PMID:25340464

Peirano, Gisele; Bradford, Patricia A.; Kazmierczak, Krystyna M.; Badal, Robert E.; Hackel, Meredith; Hoban, Daryl J.

2014-01-01

246

Biological Costs and Mechanisms of Fosfomycin Resistance in Escherichia coli  

Microsoft Academic Search

Fosfomycin is a cell wall inhibitor used mainly for the treatment of uncomplicated lower urinary tract infections. As shown here, resistance to fosfomycin develops rapidly in Escherichia coli under experimental conditions, but in spite of the relatively high mutation rate in vitro, resistance in clinical isolates is rare. To examine this apparent contradiction, we mathematically modeled the probability of resistance

Annika I. Nilsson; Otto G. Berg; Olle Aspevall; Gunnar Kahlmeter; Dan I. Andersson

2003-01-01

247

Structure and Mechanism of the Lactose Permease of Escherichia coli  

Microsoft Academic Search

Membrane transport proteins that transduce free energy stored in electrochemical ion gradients into a concentration gradient are a major class of membrane proteins. We report the crystal structure at 3.5 angstroms of the Escherichia coli lactose permease, an intensively studied member of the major facilitator superfamily of transporters. The molecule is composed of N- and C-terminal domains, each with six

Jeff Abramson; Irina Smirnova; Vladimir Kasho; Gillian Verner; H. Ronald Kaback; So Iwata

2003-01-01

248

Bactericidal activity of human serum against Escherichia coli chi1776.  

PubMed Central

Escherichia coli chi1776 is a host strain providing the EK2 level of biological containment. Laboratories equipped to use this strain may be located near medical care facilities where patients prone to infection may be found. We have therefore documented the marked susceptibility of chi1776 to killing by human serum from normal as well as ill individuals. PMID:6995323

Alexander, W J; Alexander, L S; Curtiss, R

1980-01-01

249

Recombinant protein folding and misfolding in Escherichia coli  

Microsoft Academic Search

The past 20 years have seen enormous progress in the understanding of the mechanisms used by the enteric bacterium Escherichia coli to promote protein folding, support protein translocation and handle protein misfolding. Insights from these studies have been exploited to tackle the problems of inclusion body formation, proteolytic degradation and disulfide bond generation that have long impeded the production of

Mirna Mujacic; François Baneyx

2004-01-01

250

Changes in Escherichia coli transcriptome during acclimatization at low temperature  

Microsoft Academic Search

Upon cold shock Escherichia coli transiently stops growing and adapts to the new temperature (acclimatization phase). The major physiological effects of cold temperature are a decrease in membrane fluidity and the stabilization of secondary structures of RNA and DNA, which may affect the efficiencies of translation, transcription, and replication. Specific proteins are transiently induced in the acclimatization phase. mRNA stabilization

Alessandra Polissi; Walter De Laurentis; Sandro Zangrossi; Federica Briani; Vera Longhi; Graziano Pesole; Gianni Dehò

2003-01-01

251

Quorum sensing in Escherichia coli and Salmonella typhimurium  

Microsoft Academic Search

Escherichia coli and Salmonella typhimurium strains grown in Luria-Bertani medium containing glucose secrete a small soluble heat labile organic molecule that is involved in intercellular communication. The factor is not produced when the strains are grown in Luria-Bertani me- dium in the absence of glucose. Maximal secretion of the substance occurs in midexponential phase, and the extracel- lular activity is

MICHAEL G. SURETTE; BONNIE L. BASSLER

1998-01-01

252

Genetics of ribosomal protein methylation in Escherichia coli  

Microsoft Academic Search

Two genes governing ribosomal protein methylation have been located on the map of Escherichia coli by conjugation and transduction crosses between wild-type and prm (protein methylation) mutants. The Prm phenotype of recombinants was determined by an in vitro assay of methylgroups incorporation into protein.

Charles Colson; Jacques Lhoest; Colette Urlings

1979-01-01

253

Carbon nutrition of Escherichia coli in the mouse intestine  

E-print Network

Carbon nutrition of Escherichia coli in the mouse intestine Dong-Eun Chang*, Darren J. Smalley in the mammalian intestine. Most were nutritional genes corresponding to catabolic pathways for nutrients found of the mouse intestine in competition with their wild-type parent. We found that only mutations in sugar

Conway, Tyrrell

254

Budget Analysis of Escherichia coli at a Southern Lake Michigan  

E-print Network

Budget Analysis of Escherichia coli at a Southern Lake Michigan Beach P R A M O D T H U P A K I and the Environment, University of Michigan, Ann Arbor, Michigan, NOAA Great Lakes Environmental Research Laboratory (GLERL), Ann Arbor, Michigan, and Lake Michigan Ecological Research Station, U.S. Geological Survey

255

Original article Resistance of Escherichia coli growing as biofilms  

E-print Network

Original article Resistance of Escherichia coli growing as biofilms to disinfectants C Ntsama) Summary ― The bactericidal activity of various disinfectants (cationic or amphoteric surfactants in a continuous culture system. The bacteria tested on germ-carriers or included in biofilms were more resistant

Boyer, Edmond

256

The deoxycytidine pathway for thymidylate synthesis in Escherichia coli.  

PubMed

When thymidylate production is diminished by a mutation affecting dCTP deaminase, Escherichia coli is known to use an alternate pathway involving deoxycytidine as an intermediate. The pathway requires the gene for any of three nucleoside diphosphate kinases (ndk, pykA, or pykF) and the gene for a 5'-nucleotidase (yfbR). PMID:17827303

Weiss, Bernard

2007-11-01

257

Pharmacodynamic profiling of commonly prescribed antimicrobial drugs against Escherichia coli isolates from urinary tract.  

PubMed

Since antimicrobial resistance among uropathogens against current first line agents has affected the management of severe urinary tract infection, we determined the likelihood that antibiotic regimens achieve bactericidal pharmacodynamic exposures using Monte Carlo simulation for five antimicrobials (ciprofloxacin, ceftriaxone, piperacillin/tazobactam, ertapenem, and meropenem) commonly prescribed as initial empirical treatment of inpatients with severe community acquired urinary tract infections. Minimum inhibitory concentration determination by Etest was performed for 205 Brazilian community urinary tract infection Escherichia coli strains from 2008 to 2012 and 74 E. coli bloodstream strains recovered from a surveillance study. Pharmacodynamic exposure was modeled via a 5000 subject Monte Carlo simulation. All isolates were susceptible to ertapenem and meropenem. Piperacillin/tazobactam, ceftriaxone and ciprofloxacin showed 100%, 97.5% and 83.3% susceptibility among outpatient isolates and 98.6%, 75.7% and 64.3% among inpatient isolates, respectively. Against outpatient isolates, all drugs except ciprofloxacin (82.7% in aggressive and 77.6% in conservative scenarios) achieved high cumulative fraction of response: carbapenems and piperacillin/tazobactam cumulative fraction of responses were close to 100%, and ceftriaxone cumulative fraction of response was 97.5%. Similar results were observed against inpatients isolates for carbapenems (100%) and piperacillin/tazobactam (98.4%), whereas ceftriaxone achieved only 76.9% bactericidal cumulative fraction of response and ciprofloxacin 61.9% (aggressive scenario) and 56.7% (conservative scenario) respectively. Based on this model, standard doses of beta-lactams were predicted to deliver sufficient pharmacodynamic exposure for outpatients. However, ceftriaxone should be avoided for inpatients and ciprofloxacin empirical prescription should be avoided in both inpatients and outpatients with complicated urinary tract infection. PMID:24731938

Cuba, Gabriel Trova; Pignatari, Antonio Carlos Campos; Patekoski, Katya Silva; Luchesi, Lucimila Jorge; Kiffer, Carlos Roberto Veiga

2014-01-01

258

Flow Cytometry Analysis Using Sysmex UF-1000i Classifies Uropathogens Based on Bacterial, Leukocyte, and Erythrocyte Counts in Urine Specimens among Patients with Urinary Tract Infections.  

PubMed

Urinary tract infections (UTIs) are the second most common bacterial infection. Urine culture is the gold standard for diagnosis, but new techniques, such as flow cytometry analysis (FCA), have been introduced. The aim of the present study was to evaluate FCA characteristics regarding bacteriuria, leukocyturia, and erythrocyturia in relation to cultured uropathogens in specimens from patients with a suspected UTI. We also wanted to evaluate whether the FCA characteristics can identify uropathogens prior to culture. From a prospective study, 1,587 consecutive urine specimens underwent FCA prior to culture during January and February 2012. Outpatients and inpatients (79.6% and 19.4%, respectively) were included, of whom women represented 67.5%. In total, 620 specimens yielded growth, of which Escherichia coli represented 65%, Enterococcus spp. 8%, Klebsiella spp. 7%, and Staphylococcus spp. 5%. For the uropathogens, the outcome of FCA was compared against the results for specimens with E. coli and those with a negative culture. E. coli had high bacterial (median, 17,914/?l), leukocyte (median, 348/?l), and erythrocyte (median, 23/?l) counts. With the exception of Klebsiella spp., the majority of the uropathogens had considerable or significantly lower bacterial counts than that of E. coli. High leukocyte counts were found in specimens with Staphylococcus aureus, Proteus mirabilis, Pseudomonas aeruginosa, and group C streptococci. Elevated erythrocyte counts were found for P. vulgaris, P. aeruginosa, and group C streptococci, as well as for Staphylococcus saprophyticus. In essence, FCA adds new information about the bacterial, leukocyte, and erythrocyte counts in urine specimens for different uropathogens. Based on FCA characteristics, uropathogens can be classified and identified prior to culture. E. coli and Klebsiella spp. have similar FCA characteristics. PMID:25472486

Monsen, Tor; Rydén, Patrik

2015-02-01

259

Kinetics of Bacteriophage ? Deoxyribonucleic Acid Infection of Escherichia coli  

PubMed Central

Barnhart, Benjamin J. (Los Alamos Scientific Laboratory, University of California, Los Alamos, N.M.). Kinetics of bacteriophage ? deoxyribonucleic acid infection of Escherichia coli. J. Bacteriol. 90:1617–1623. 1965.—The kinetics of Escherichia coli K-12 infection by phage ? deoxyribonucleic acid (DNA) were determined. An initial lag of 55 to 80 sec was found to be the time required for infecting DNA to become deoxyribonuclease-insensitive at 33 C. When cell-DNA interactions were stopped by washing away unbound DNA, the already bound DNA continued to infect the cell at rates described by linear kinetics with no apparent lag. Whereas the lag period was relatively insensitive to DNA and cell concentrations, both the lag and the subsequent linear portions of the rate curves were temperature-sensitive. Cell and DNA dose-response curves prescribed hyperbolic functions. Similarities between ? DNA infection of E. coli and bacterial transformation systems are discussed. PMID:5322721

Barnhart, Benjamin J.

1965-01-01

260

Uropathogenic Escherichia coli Superinfection Enhances the Severity of Mouse Bladder Infection.  

PubMed

Urinary tract infections (UTIs) afflict over 9 million women in America every year, often necessitating long-term prophylactic antibiotics. One risk factor for UTI is frequent sexual intercourse, which dramatically increases the risk of UTI. The mechanism behind this increased risk is unknown; however, bacteriuria increases immediately after sexual intercourse episodes, suggesting that physical manipulation introduces periurethral flora into the urinary tract. In this paper, we investigated whether superinfection (repeat introduction of bacteria) resulted in increased risk of severe UTI, manifesting as persistent bacteriuria, high titer bladder bacterial burdens and chronic inflammation, an outcome referred to as chronic cystitis. Chronic cystitis represents unchecked luminal bacterial replication and is defined histologically by urothelial hyperplasia and submucosal lymphoid aggregates, a histological pattern similar to that seen in humans suffering chronic UTI. C57BL/6J mice are resistant to chronic cystitis after a single infection; however, they developed persistent bacteriuria and chronic cystitis when superinfected 24 hours apart. Elevated levels of interleukin-6 (IL-6), keratinocyte cytokine (KC/CXCL1), and granulocyte colony-stimulating factor (G-CSF) in the serum of C57BL/6J mice prior to the second infection predicted the development of chronic cystitis. These same cytokines have been found to precede chronic cystitis in singly infected C3H/HeN mice. Furthermore, inoculating C3H/HeN mice twice within a six-hour period doubled the proportion of mice that developed chronic cystitis. Intracellular bacterial replication, regulated hemolysin (HlyA) expression, and caspase 1/11 activation were essential for this increase. Microarrays conducted at four weeks post inoculation in both mouse strains revealed upregulation of IL-1 and antimicrobial peptides during chronic cystitis. These data suggest a mechanism by which caspase-1/11 activation and IL-1 secretion could predispose certain women to recurrent UTI after frequent intercourse, a predisposition predictable by several serum biomarkers in two murine models. PMID:25569799

Schwartz, Drew J; Conover, Matt S; Hannan, Thomas J; Hultgren, Scott J

2015-01-01

261

Intrauterine Growth Restriction Is a Direct Consequence of Localized Maternal Uropathogenic Escherichia coli Cystitis  

PubMed Central

Despite the continually increasing rates of adverse perinatal outcomes across the globe, the molecular mechanisms that underlie adverse perinatal outcomes are not completely understood. Clinical studies report that 10% of pregnant women will experience a urinary tract infection (UTI) and there is an association of UTIs with adverse perinatal outcomes. We introduced bacterial cystitis into successfully outbred female mice at gestational day 14 to follow pregnancy outcomes and immunological responses to determine the mechanisms that underlie UTI-mediated adverse outcomes. Outbred fetuses from mothers experiencing localized cystitis displayed intrauterine growth restriction (20–80%) as early as 48 hours post-infection and throughout the remainder of normal gestation. Robust infiltration of cellular innate immune effectors was observed in the uteroplacental tissue following introduction of UTI despite absence of viable bacteria. The magnitude of serum proinflammatory cytokines is elevated in the maternal serum during UTI. This study demonstrates that a localized infection can dramatically impact the immunological status as well as the function of non-infected distal organs and tissues. This model can be used as a platform to determine the mechanism(s) by which proinflammatory changes occur between non-contiguous genitourinary organs PMID:22470490

Bolton, Michael; Horvath, Dennis J.; Li, Birong; Cortado, Hanna; Newsom, David; White, Peter; Partida-Sanchez, Santiago; Justice, Sheryl S.

2012-01-01

262

Uropathogenic Escherichia coli Superinfection Enhances the Severity of Mouse Bladder Infection  

PubMed Central

Urinary tract infections (UTIs) afflict over 9 million women in America every year, often necessitating long-term prophylactic antibiotics. One risk factor for UTI is frequent sexual intercourse, which dramatically increases the risk of UTI. The mechanism behind this increased risk is unknown; however, bacteriuria increases immediately after sexual intercourse episodes, suggesting that physical manipulation introduces periurethral flora into the urinary tract. In this paper, we investigated whether superinfection (repeat introduction of bacteria) resulted in increased risk of severe UTI, manifesting as persistent bacteriuria, high titer bladder bacterial burdens and chronic inflammation, an outcome referred to as chronic cystitis. Chronic cystitis represents unchecked luminal bacterial replication and is defined histologically by urothelial hyperplasia and submucosal lymphoid aggregates, a histological pattern similar to that seen in humans suffering chronic UTI. C57BL/6J mice are resistant to chronic cystitis after a single infection; however, they developed persistent bacteriuria and chronic cystitis when superinfected 24 hours apart. Elevated levels of interleukin-6 (IL-6), keratinocyte cytokine (KC/CXCL1), and granulocyte colony-stimulating factor (G-CSF) in the serum of C57BL/6J mice prior to the second infection predicted the development of chronic cystitis. These same cytokines have been found to precede chronic cystitis in singly infected C3H/HeN mice. Furthermore, inoculating C3H/HeN mice twice within a six-hour period doubled the proportion of mice that developed chronic cystitis. Intracellular bacterial replication, regulated hemolysin (HlyA) expression, and caspase 1/11 activation were essential for this increase. Microarrays conducted at four weeks post inoculation in both mouse strains revealed upregulation of IL-1 and antimicrobial peptides during chronic cystitis. These data suggest a mechanism by which caspase-1/11 activation and IL-1 secretion could predispose certain women to recurrent UTI after frequent intercourse, a predisposition predictable by several serum biomarkers in two murine models. PMID:25569799

Schwartz, Drew J.; Conover, Matt S.; Hannan, Thomas J.; Hultgren, Scott J.

2015-01-01

263

The affinity of the FimH fimbrial adhesin is receptor-driven and quasi-independent of Escherichia coli pathotypes  

PubMed Central

Type-1 fimbriae are important virulence factors for the establishment of Escherichia coli urinary tract infections. Bacterial adhesion to the high-mannosylated uroplakin Ia glycoprotein receptors of bladder epithelium is mediated by the FimH adhesin. Previous studies have attributed differences in mannose-sensitive adhesion phenotypes between faecal and uropathogenic E. coli to sequence variation in the FimH receptor-binding domain. We find that FimH variants from uropathogenic, faecal and enterohaemorrhagic isolates express the same specificities and affinities for high-mannose structures. The only exceptions are FimHs from O157 strains that carry a mutation (Asn135Lys) in the mannose-binding pocket that abolishes all binding. A high-mannose microarray shows that all substructures are bound by FimH and that the largest oligomannose is not necessarily the best binder. Affinity measurements demonstrate a strong preference towards oligomannosides exposing Man?1-3Man at their non-reducing end. Binding is further enhanced by the ?1-4-linkage to GlcNAc, where binding is 100-fold better than that of ?-d-mannose. Man?1-3Man?1-4GlcNAc, a major oligosaccharide present in the urine of ?-mannosidosis patients, thus constitutes a well-defined FimH epitope. Differences in affinities for high-mannose structures are at least 10-fold larger than differences in numbers of adherent bacteria between faecal and uropathogenic strains. Our results imply that the carbohydrate expression profile of targeted host tissues and of natural inhibitors in urine, such as Tamm-Horsfall protein, are stronger determinants of adhesion than FimH variation. PMID:16930149

Bouckaert, Julie; Mackenzie, Jenny; de Paz, José L; Chipwaza, Beatrice; Choudhury, Devapriya; Zavialov, Anton; Mannerstedt, Karin; Anderson, Jennifer; Piérard, Denis; Wyns, Lode; Seeberger, Peter H; Oscarson, Stefan; De Greve, Henri; Knight, Stefan D

2006-01-01

264

Novel phage-based bio-processing of pathogenic Escherichia coli and its biofilms  

Microsoft Academic Search

To explore new approaches of phage-based bio-process of specifically pathogenic Escherichia coli bacteria in food products within a short period. One hundred and forty highly lytic designed coliphages were used. Escherichia coli naturally contaminated and Enterohemorrhagic Escherichia coli experimentally inoculated samples of lettuce, cabbage, meat, and egg were used. In addition, experimentally produced biofilms\\u000a of E. coli were tested. A

S. A. A. Jassim; A. S. Abdulamir; F. Abu Bakar

265

Lytic bacteriophages reduce Escherichia coli O157  

PubMed Central

The role of lytic bacteriophages in preventing cross contamination of produce has not been evaluated. A cocktail of three lytic phages specific for E. coli O157:H7 (EcoShield™) or a control (phosphate buffered saline, PBS) was applied to lettuce by either; (1) immersion of lettuce in 500 ml of EcoShield™ 8.3 log PFU/ml or 9.8 log PFU/ml for up to 2 min before inoculation with E. coli O157:H7; (2) spray-application of EcoShield™ (9.3 log PFU/ml) to lettuce after inoculation with E. coli O157:H7 (4.10 CFU/cm2) following exposure to 50 ?g/ml chlorine for 30 sec. After immersion studies, lettuce was spot-inoculated with E. coli O157:H7 (2.38 CFU/cm2). Phage-treated, inoculated lettuce pieces were stored at 4°C for and analyzed for E. coli O157:H7 populations for up to 7 d. Immersion of lettuce in 9.8 log PFU/ml EcoShield™ for 2 min significantly (p < 0.05) reduced E. coli O157:H7 populations after 24 h when stored at 4°C compared with controls. Immersion of lettuce in suspensions containing high concentrations of EcoShield™ (9.8 log PFU/ml) resulted in the deposition of high concentrations (7.8 log log PFU/cm2) of bacteriophages on the surface of fresh cut lettuce, potentially contributing to the efficacy of the lytic phages on lettuce. Spraying phages on to inoculated fresh cut lettuce after being washed in hypochlorite solution was significantly more effective in reducing E. coli O157:H7 populations (2.22 log CFU/cm2) on day 0 compared with control treatments (4.10 log CFU/cm2). Both immersion and spray treatments provided protection from E. coli O157:H7 contamination on lettuce, but spray application of lytic bacteriophages to lettuce was more effective in immediately reducing E. coli O157:H7 populations fresh cut lettuce. PMID:23819106

Ferguson, Sean; Roberts, Cheryl; Handy, Eric; Sharma, Manan

2013-01-01

266

First report on class 1 integrons and Trimethoprim-resistance genes from dfrA group in uropathogenic E. coli (UPEC) from the Aleppo area in Syria  

PubMed Central

Horizontal gene transfer (HGT) introduces advantageous genetic elements into pathogenic bacteria using tools such as class1 integrons. This study aimed at investigating the distribution of these integrons among uropathogenic E. coli (UPEC) isolated from patients in Aleppo, Syria. It also set to uncover the frequencies of the clinically relevant DfrA1 and DfrA17,7, as well as various associations leading to reduced susceptibility. This study involved 75 Trimethoprim-resistant E. coli isolates from in- and outpatients with urinary tract infections (UTIs) from 3 major hospitals in Aleppo. Bacterial identification, resistance and extended-spectrum-?-lactamase (ESBL) production testing were performed according to Clinical Laboratory Standards Institute guidelines. Detection of integrons and DfrA genes was done using PCR and statistical significance was inferred through ?2 (Fisher’s) test. Class1 integrons were detected in 54.6% of isolates while DfrA1 and DfrA17,7 were found in 16% and 70.6% of tested samples respectively. Furthermore, only DfrA17,7 were strongly associated with class1 integrons, as were reduced susceptibility to the majority of individual antibiotics, multidrug resistance and ESBL production. This study demonstrated the high prevalence of class1 integrons among UPEC strains in Aleppo, Syria, as well as their significant associations with MDR. This data give information for local healthcare provision using antibiotic chemotherapy. PMID:23956949

Al-Assil, Bodour; Mahfoud, Maysa; Hamzeh, Abdul Rezzak

2013-01-01

267

[Acute diarrheal disease caused by enteropathogenic Escherichia coli in Colombia].  

PubMed

Intestinal Escherichia coli pathogens are leading causes of acute diarrheal disease in children less than 5 years in Latin America, Africa and Asia and a leading cause of death in children living in poorest communities in Africa and South East Asia. Studies on the role of E. coli pathogens in childhood diarrhea in Colombia and other countries in Latin America are limited due to the lack of detection assays in clinical laboratories at the main urban medical centers. Recent studies report that enterotoxigenic E. coli is the most common E. coli pathogens associated with diarrhea in children less than 5 years of age. Other E. coli pathotypes have been detected in children with diarrhea including enteropathogenic, enteroaggregative, shiga-toxin producing and diffusely adherent E. coli. It was also found that meat and vegetables at retail stores are contaminated with Shiga-toxin producing E. coli and enteroaggregative E. coli, suggesting that food products are involved in transmission and infection of the susceptible host. More studies are necessary to evaluate the mechanisms of transmission, the impact on the epidemiology of diarrheal disease, and management strategies and prevention of these pathogens affecting the pediatric population in Colombia. PMID:25491457

Gómez-Duarte, Oscar G

2014-10-01

268

Biosynthesis of bioactive O-methylated flavonoids in Escherichia coli.  

PubMed

Two bioactive O-methylflavonoids, sakuranetin (7-O-methylnaringenin) and ponciretin (7-O-methylnaringenin), were synthesized in Escherichia coli. Sakuranetin inhibits germination of Magnaporthe grisea, and ponciretin is a potential inhibitor of Helicobacter pylori. To achieve this, we reconstructed the naringenin biosynthesis pathway in E. coli. First, the shikimic acid pathway, which leads to the biosynthesis of tyrosine, was engineered in E. coli to increase the amount of available tyrosine. Second, several genes for the biosynthesis of ponciretin and sakuranetin such as tyrosine ammonia lyase (TAL), 4-coumaroyl CoA ligase (4CL), chalcone synthase (CHS), and O-methyltransferase (OMT) were overexpressed. In order to increase the supply the Coenzyme A (CoA), one gene (icdA, isocitrate dehydrogenase) was deleted. Using these strategies, we synthesized ponciretin and sakuranetin from glucose in E. coli at the concentration of 42.5 mg/L and 40.1 mg/L, respectively. PMID:23771780

Kim, Min-Ji; Kim, Bong-Gyu; Ahn, Joong-Hoon

2013-08-01

269

Adhesion of Human and Animal Escherichia coli Strains in Association with Their Virulence-Associated Genes and Phylogenetic Origins  

PubMed Central

Intestinal colonization is influenced by the ability of the bacterium to inhabit a niche, which is based on the expression of colonization factors. Escherichia coli carries a broad range of virulence-associated genes (VAGs) which contribute to intestinal (inVAGs) and extraintestinal (exVAGs) infection. Moreover, initial evidence indicates that inVAGs and exVAGs support intestinal colonization. We developed new screening tools to genotypically and phenotypically characterize E. coli isolates originating in humans, domestic pigs, and 17 wild mammal and avian species. We analyzed 317 isolates for the occurrence of 44 VAGs using a novel multiplex PCR microbead assay (MPMA) and for adhesion to four epithelial cell lines using a new adhesion assay. We correlated data for the definition of new adhesion genes. inVAGs were identified only sporadically, particularly in roe deer (Capreolus capreolus) and the European hedgehog ( Erinaceus europaeus). The prevalence of exVAGs depended on isolation from a specific host. Human uropathogenic E. coli isolates carried exVAGs with the highest prevalence, followed by badger (Meles meles) and roe deer isolates. Adhesion was found to be very diverse. Adhesion was specific to cells, host, and tissue, though it was also unspecific. Occurrence of the following VAGs was associated with a higher rate of adhesion to one or more cell lines: afa-dra, daaD, tsh, vat, ibeA, fyuA, mat, sfa-foc, malX, pic, irp2, and papC. In summary, we established new screening methods which enabled us to characterize large numbers of E. coli isolates. We defined reservoirs for potential pathogenic E. coli. We also identified a very broad range of colonization strategies and defined potential new adhesion genes. PMID:23872574

Frömmel, Ulrike; Lehmann, Werner; Rödiger, Stefan; Böhm, Alexander; Nitschke, Jörg; Weinreich, Jörg; Groß, Julia; Roggenbuck, Dirk; Zinke, Olaf; Ansorge, Hermann; Vogel, Steffen; Klemm, Per; Wex, Thomas; Schröder, Christian; Wieler, Lothar H.

2013-01-01

270

BIOSYNTHESIS AND BIOSYNTHETIC PATHWAYS OF PENTOSES IN ESCHERICHIA COLI  

PubMed Central

Sable, Henry Z. (Western Reserve University, Cleveland, Ohio) and Elayne E. Cassisi. Biosynthesis and biosynthetic pathways of pentoses in Escherichia coli. J. Bacteriol. 84:1169–1172. 1962.—Resting glucose-adapted Escherichia coli supplied with glucose continues to synthesize pentose by the oxidative pathway characteristic of logarithmically growing glucose-adapted cells. This behavior is unlike that of acetate-adapted resting E. coli supplied with glucose, which continues to synthesize most of its pentose by the nonoxidative pathway characteristic of acetate-adapted cells. When infected with bacteriophage T2H, E. coli continues to use the oxidative pathway predominantly. This finding is in contrast to reports that infection with T6r+ bacteriophage increases the participation of a nonoxidative pathway. Resting glucose-adapted E. coli supplied with acetate-1-C14 as sole carbon source synthesizes pentose by a pathway or pathways which cannot be assessed completely by methods previously developed (which are based on the relative labeling of C-1, C-2, and C-3 of the pentose) but which is most probably predominantly nonoxidative. PMID:13975889

Sable, Henry Z.; Cassisi, Elayne E.

1962-01-01

271

Uncoupler Resistance in Escherichia coli: the Role of Cellular Respiration  

Microsoft Academic Search

Bioenergetic properties of a mutant strain of Escherichia coli K12 designated TUV, which is resistant to the protonophoric uncoupling agent 4,5,6,7-tetrachloro-2-trifluoromethylbenzimid- azole (TTFB) have been compared with those of its non-resistant parent, E. coli K12 Doc-S. Strain TUV grew and respired some 20-30% faster than strain Doc-S, and was cross-resistant to carbonylcyanide p-(trifluoromethoxy)phenylhydrazone and triphenyltin, but not to 2,4- dinitrophenol.

PHILIP G. QUIRK; MICHAEL R. JONES; ROBERT S. HAWORTH; R. BRIAN BEECHEY; IAIN D. CAMPBELL

1989-01-01

272

[Reactive arthritis due to Escherichia coli urinary tract infection].  

PubMed

Reactive arthritis following Escherichia coli urinary tract infection is very rare. We report a 25-year-old woman with acute oligoarthritis associated with bilateral anterior uveitis after an episode of urinary tract infection due to E. coli. The diagnosis of reactive arthritis was considered and the patient treated with non-steroidal anti-inflammatory agents. Disease course was rapidly successful and at 6-month follow-up the patient was asymptomatic. Reactive arthritis is associated with intestinal infection but also with common urinary tract infection. PMID:20605282

Renou, F; Wartel, G; Raffray, L; Kuli, B; Fayeulle, S; Yvin, J-L

2011-01-01

273

Functional Role of bdm During Flagella Biogenesis in Escherichia coli.  

PubMed

The biofilm-dependent modulation gene (bdm) has recently been shown to play a role in osmotic-induced formation of biofilm in Escherichia coli. In this study, we demonstrated that deletion of bdm results in down-regulation of flagella biosynthesis genes and, consequently, a defect in E. coli motility. In addition, we employed atomic force microscopy to confirm the absence of flagella-like structures on the surface of bdm-null cells. These findings indicate that bdm plays a key role in regulatory pathway for the formation of flagella. PMID:25398323

Kim, Ji-Sun; Kim, Yu Jin; Seo, Sojin; Seong, Maeng-Je; Lee, Kangseok

2015-03-01

274

Inactivation of Escherichia coli using atmospheric-pressure plasma jet  

NASA Astrophysics Data System (ADS)

An atmospheric-pressure argon (Ar) plasma jet was applied to the inactivation of Escherichia coli. The Ar plasma jet was generated at a frequency of 10 kHz, an applied voltage of 10 kV, and an Ar gas flow rate of 10 L/min at atmospheric pressure. E. coli cells seeded on an agar medium in a Petri dish were inactivated by Ar plasma jet irradiation for 1 s. Scanning electron microscopy (SEM) revealed that E. coli cells were killed because their cell wall and membrane were disrupted. To determine the causes of the disruption of the cell wall and membrane of E. coli, we performed the following experiments: the measurement of the surface temperature of an agar medium using a thermograph, the analysis of an emission spectrum of a plasma jet obtained using a multichannel spectrometer, and the determination of the distribution of the concentration of hydrogen peroxide (H2O2) generated on an agar medium by plasma jet irradiation using semiquantitative test strips. Moreover, H2O2 solutions of different concentrations were dropped onto an agar medium seeded with E. coli cells to examine the contribution of H2O2 to the death of E. coli. The results of these experiments showed that the cell wall and membrane of E. coli were disrupted by electrons in the plasma jet, as well as by electroneutral excited nitrogen molecules (N2) and hydroxyl (OH) radicals in the periphery of the plasma jet.

Kuwahata, Hiroshi; Yamaguchi, Takeshi; Ohyama, Ryu-ichiro; Ito, Atsushi

2015-01-01

275

Determination and quantification of Escherichia coli by capillary electrophoresis.  

PubMed

Capillary electrophoresis (CE) is widely employed for the separation of nucleic acids or protein, but it is rarely applied in the quantification of Escherichia coli (E. coli). Here, we have analysed E. coli by CE with mercury lamp induced fluorescence, and demonstrated the relationship between its fluorescence intensity with the concentration of E. coli for the first time. The gradient concentration of E. coli was obtained by polymerase chain reaction (PCR) with different amplification cycles and dilution certain PCR products of E. coli, respectively. Results show that the peak area was linearly related to the logarithm of the concentration of E. coli and the logarithm of PCR replication numbers. The correlation coefficients R(2) are 0.957 and 0.966, respectively. The limit of detection (LOD) was found to be about 8.913 × 10(-15) mol ?l(-1). The reproducibility of capillary electrophoresis may make this technique possible for quantitative measurement of bacteria in bio-analytical science. PMID:25307062

Li, Zhenqing; Li, De; Zhang, Dawei; Yamaguchi, Yoshinori

2014-12-01

276

Enhancing the Antibiotic Antibacterial Effect by Sub Lethal Tellurite Concentrations: Tellurite and Cefotaxime Act Synergistically in Escherichia coli  

PubMed Central

The emergence of antibiotic-resistant pathogenic bacteria during the last decades has become a public health concern worldwide. Aiming to explore new alternatives to treat antibiotic-resistant bacteria and given that the tellurium oxyanion tellurite is highly toxic for most microorganisms, we evaluated the ability of sub lethal tellurite concentrations to strengthen the effect of several antibiotics. Tellurite, at nM or µM concentrations, increased importantly the toxicity of defined antibacterials. This was observed with both Gram negative and Gram positive bacteria, irrespective of the antibiotic or tellurite tolerance of the particular microorganism. The tellurite-mediated antibiotic-potentiating effect occurs in laboratory and clinical, uropathogenic Escherichia coli, especially with antibiotics disturbing the cell wall (ampicillin, cefotaxime) or protein synthesis (tetracycline, chloramphenicol, gentamicin). In particular, the effect of tellurite on the activity of the clinically-relevant, third-generation cephalosporin (cefotaxime), was evaluated. Cell viability assays showed that tellurite and cefotaxime act synergistically against E. coli. In conclusion, using tellurite like an adjuvant could be of great help to cope with several multi-resistant pathogens. PMID:22536386

Molina-Quiroz, Roberto C.; Muñoz-Villagrán, Claudia M.; de la Torre, Erick; Tantaleán, Juan C.; Vásquez, Claudio C.; Pérez-Donoso, José M.

2012-01-01

277

Enterotoxigenic Escherichia coli infection in captive black-footed ferrets.  

PubMed

Enterotoxigenic Escherichia coli with genes for heat stabile toxins Sta and STb was isolated from the gastrointestinal tract and multiple visceral organs of three adult and three juvenile black-footed ferrets (Mustela nigripes) that died in a captive breeding colony between 24 May 1998 and 2 July 1998. Similar isolates were obtained from rectal swabs of one adult and one juvenile that were clinically ill. All were fed a diet composed of mink chow, raw rabbit meat, beef liver powder, blood meal and lard. Escherichia coli of the same toxin genotype was isolated from the mixed ration. Clinical signs included sudden death, dehydration, anorexia and diarrhea. Necropsy lesions included acute enteritis with large numbers of rod shaped bacteria microscopically visible on intestinal villi. PMID:11504237

Bradley, G A; Orr, K; Reggiardo, C; Glock, R D

2001-07-01

278

Expression of soluble cloned porcine pepsinogen A in Escherichia coli.  

PubMed Central

A system for the production of soluble porcine pepsinogen A (EC 3.4.23.1) was developed by fusing the pepsinogen and thioredoxin genes and then expressing the fused product (Trx-PG) in Escherichia coli. The expressed fusion protein was purified using a combination of ion-exchange and hydrophobic chromatography. Trypsin digestion of the fusion protein yielded pepsinogen which was one residue longer than the intrinsic length. Acidification of either the fusion protein or pepsinogen (tryptic digestion of Trx-PG) yielded recombinant pepsin A (r-pepsin). When compared with commercial porcine pepsin A, r-pepsin had similar milk-clotting and proteolytic activities, kinetic parameters and pH dependency. The above results indicate that an expression system was developed which yielded fully active soluble pepsin(ogen) from Escherichia coli. PMID:8615812

Tanaka, T; Yada, R Y

1996-01-01

279

Development of functionalised polyelectrolyte capsules using filamentous Escherichia coli cells  

PubMed Central

Background Escherichia coli is one of the best studied microorganisms and finds multiple applications especially as tool in the heterologous production of interesting proteins of other organisms. The heterologous expression of special surface (S-) layer proteins caused the formation of extremely long E. coli cells which leave transparent tubes when they divide into single E. coli cells. Such natural structures are of high value as bio-templates for the development of bio-inorganic composites for many applications. In this study we used genetically modified filamentous Escherichia coli cells as template for the design of polyelectrolyte tubes that can be used as carrier for functional molecules or particles. Diversity of structures of biogenic materials has the potential to be used to construct inorganic or polymeric superior hybrid materials that reflect the form of the bio-template. Such bio-inspired materials are of great interest in diverse scientific fields like Biology, Chemistry and Material Science and can find application for the construction of functional materials or the bio-inspired synthesis of inorganic nanoparticles. Results Genetically modified filamentous E. coli cells were fixed in 2% glutaraldehyde and coated with alternating six layers of the polyanion polyelectrolyte poly(sodium-4styrenesulfonate) (PSS) and polycation polyelectrolyte poly(allylamine-hydrochloride) (PAH). Afterwards we dissolved the E. coli cells with 1.2% sodium hypochlorite, thus obtaining hollow polyelectrolyte tubes of 0.7 ?m in diameter and 5–50 ?m in length. For functionalisation the polyelectrolyte tubes were coated with S-layer protein polymers followed by metallisation with Pd(0) particles. These assemblies were analysed with light microscopy, scanning electron microscopy, energy dispersive X-ray spectroscopy and transmission electron microscopy. Conclusion The thus constructed new material offers possibilities for diverse applications like novel catalysts or metal nanowires for electrical devices. The novelty of this work is the use of filamentous E. coli templates and the use of S-layer proteins in a new material construct. PMID:23259586

2012-01-01

280

Escherichia coli and Salmonella 2000: the view from here.  

PubMed

Five years after the publication of the second edition of the reference book Escherichia coli and Salmonella: Cellular and Molecular Biology, and on the eve of launching a successor venture, the editors and colleagues examine where we stand in our quest for an understanding of these organisms. The main areas selected for this brief inquiry are genomics, evolution, molecular multifunctionality, functional backups, regulation of gene expression, cell biology, sensing of the environment, and ecology. PMID:11238988

Schaechter, M

2001-03-01

281

Escherichia coli acid resistance: tales of an amateur acidophile  

Microsoft Academic Search

Gastrointestinal pathogens are faced with an extremely acidic environment. Within moments, a pathogen such as Escherichia coli O157:H7 can move from the nurturing pH 7 environment of a hamburger to the harsh pH 2 milieu of the stomach. Surprisingly, certain microorganisms that grow at neutral pH have elegantly regulated systems that enable survival during excursions into acidic environments. The best-characterized

John W. Foster

2004-01-01

282

Expression of the Human Erythrocyte Glucose Transporter in Escherichia coli  

Microsoft Academic Search

The gene encoding the human erythrocyte glucose transporter, cloned from HepG2 hepatoma cells, was expressed in Escherichia coli by introducing a prokaryote-type ribosome binding site, subcloning the gene into the T7 promoter\\/T7 polymerase expression system, and transforming a strain that is defective in glucose transport. Cells bearing plasmids with the transporter gene take up 2-deoxy-D-glucose and D-glucose, unlike cells bearing

Hemanta K. Sarkar; Bernard Thorens; Harvey F. Lodish; H. Ronald Kaback

1988-01-01

283

Electric field induced bacterial flocculation of enteroaggregative Escherichia coli 042  

NASA Astrophysics Data System (ADS)

A response of the aggregation dynamics of enteroaggregative Escherichia coli under low magnitude steady and oscillating electric fields is presented. The presence of uniform electric fields hampered microbial adhesion and biofilm formation on a transverse glass surface, but instead promoted the formation of flocs. Extremely heterogenous distribution of live and dead cells was observed among the flocs. Moreover, floc formation was largely observed to be independent of the frequency of alternating electric fields.

Kumar, Aloke; Mortensen, Ninell P.; Mukherjee, Partha P.; Retterer, Scott T.; Doktycz, Mitchel J.

2011-06-01

284

Effect of cobalt on Escherichia coli metabolism and metalloporphyrin formation  

Microsoft Academic Search

Toxicity in Escherichia coli resulting from high concentrations of cobalt has been explained by competition of cobalt with iron in various metabolic processes\\u000a including Fe–S cluster assembly, sulfur assimilation, production of free radicals and reduction of free thiol pool. Here we\\u000a present another aspect of increased cobalt concentrations in the culture medium resulting in the production of cobalt protoporphyrin\\u000a IX

Tomas Majtan; Frank E. Frerman; Jan P. Kraus

2011-01-01

285

Polymorphous crystallization and diffraction of threonine deaminase from Escherichia coli.  

PubMed

The biosynthetic threonine deaminase from Escherichia coli, an allosteric tetramer with key regulatory functions, has been crystallized in several crystal forms. Two distinct forms, both belonging to either space group P3121 or P3221, with different sized asymmetric units that both contain a tetramer, grow under identical conditions. Diffraction data sets to 2.8 A resolution (native) and 2. 9 A resolution (isomorphous uranyl derivative) have been collected from a third crystal form in space group I222. PMID:9761930

Gallagher, D T; Eisenstein, E; Fisher, K E; Zondlo, J; Chinchilla, D; Yu, H D; Dill, J; Winborne, E; Ducote, K; Xiao, G; Gilliland, G L

1998-05-01

286

Solar radiation induces sublethal injury in Escherichia coli in seawater.  

PubMed Central

Sublethal injury was noted in Escherichia coli after cells were exposed to solar radiation. Injury was detected by differential plate counts between complete and minimal media that were observed with sunlight-exposed cells but not with cells kept in the dark. Since addition of catalase or pyruvate to minimal medium overcame or repaired this injury, the catalase system appeared to be the site of injury. PMID:7013708

Kapuscinski, R B; Mitchell, R

1981-01-01

287

Depression of adenosylmethionine content of Escherichia coli by thioglycerol.  

PubMed Central

The mechanism of growth inhibition of Escherichia coli by thioglycerol was probed. The extent of growth inhibition depended on the ratio of thioglycerol to initial cell concentration. Exogenously added methionine and several methionine analogues reversed the inhibition of 2 to 40 mM thioglycerol. Exposure to thioglycerol elevated the intracellular concentration of methionine, but the level of S-adenosylmethionine was depressed. Thioglycerol was methylated in vivo to 3-methylthio-1,2-propanediol. PMID:6362560

Javor, G T

1983-01-01

288

Transcriptional Regulation of the esp Genes of Enterohemorrhagic Escherichia coli  

Microsoft Academic Search

We have determined that the genes encoding the secreted proteins EspA, EspD, and EspB of enterohemor- rhagic Escherichia coli (EHEC) are organized in a single operon. The esp operon is controlled by a promoter located 94 bp upstream from the ATG start codon of the espA gene. The promoter is activated in the early logarithmic growth phase, upon bacterial contact

FABRIZIO BELTRAMETTI; ANDREAS U. KRESSE; CARLOS A. GUZMAN

1999-01-01

289

Occurrence of Escherichia coli in Wild Cottontail Rabbits.  

PubMed

Free-ranging cottontail rabbits (Sylvilagus floridanus) from two areas in central Pennsylvania were sampled over a 4-year period. Large numbers of coliforms were isolated from the intestinal tracts of these animals; in 136 of the 141 rabbits sampled, Escherichia coli was found to be a major component of the alimentary flora. Four serogroups (O7, O77, O73, and O103) were predominant among the isolates and were considered resistant coliflora of this species of cottontail rabbit. PMID:16345208

Kozlowski, R; Glantz, P J; Anthony, R G

1977-03-01

290

Genome sequence of enterohaemorrhagic Escherichia coli O157:H7  

Microsoft Academic Search

The bacterium Escherichia coli O157:H7 is a worldwide threat to public health and has been implicated in many outbreaks of haemorrhagic colitis, some of which included fatalities caused by haemolytic uraemic syndrome. Close to 75,000 cases of O157:H7 infection are now estimated to occur annually in the United States. The severity of disease, the lack of effective treatment and the

Nicole T. Perna; Guy Plunkett; Valerie Burland; Bob Mau; Jeremy D. Glasner; Debra J. Rose; George F. Mayhew; Peter S. Evans; Jason Gregor; Heather A. Kirkpatrick; György Pósfai; Jeremiah Hackett; Sara Klink; Adam Boutin; Ying Shao; Leslie Miller; Erik J. Grotbeck; N. Wayne Davis; Alex Lim; Eileen T. Dimalanta; Konstantinos D. Potamousis; Jennifer Apodaca; Thomas S. Anantharaman; Jieyi Lin; Galex Yen; David C. Schwartz; Rodney A. Welch; Frederick R. Blattner

2001-01-01

291

Functional requirements for heat induced genome amplification in Escherichia coli  

Microsoft Academic Search

A temperature shift-up induces extra rounds of fully replicated chromosomes in Escherichia coli and leads to an increase in DNA\\/mass ratio. In the present work we characterize the requirements for this heat-induced replication (HIR) with respect to replisome components, replication restart, and recombination functions. We found that HIR requires Klenow and 5?–3? exonuclease activities from Pol I and Pol III,

Rocío González-Soltero; Alfonso Jiménez-Sánchez; Emilia Botello

2008-01-01

292

Escherichia coli as a pathogen in dogs and cats.  

PubMed

Certain strains of Escherichia coli behave as pathogens in dogs and cats causing gastro-intestinal and extra-intestinal diseases. Among the five known groups of diarrhoeagenic E. coli, namely enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), shiga-toxin producing E. coli (STEC) and enteroaggregative E. coli (EAggEC), only EPEC and ETEC were clearly associated with enteric disease in young dogs. ETEC isolates from diarrhoeic dogs were found to be positive for the heat-stable enterotoxins STa and STb but negative for heat-labile enterotoxin (LT). Canine ETEC were found to be different from those of other animals and humans by their serotypes, production of alpha-haemolysin and adhesive factors and by the production of uncharacterized types of enterotoxins by some ETEC. Canine EPEC could be distinguished from EPEC of humans or other animals by their serotypes and by the eae-protein intimin which mediates intimate adherence of EPEC to intestinal mucosa cells. STEC were occasionally isolated from faeces of healthy and diarrhoeic dogs but their role in canine diarrhoea is not yet well known. EIEC and EAggEC were not reported to occur in dogs or cats. Very little is known on diarrhoegenic E. coli in cats and further epidemiological investigations on this subject are needed. Besides its role in gastro-intestinal infections, E. coli can cause infections of the urogenital tract and systemic disease in dogs and cats. Extra-intestinal pathogenic E. coli strains from dogs and cats belong to a limited number of serotypes and clonal groups and are frequently found as a part of the normal gut flora of these animals. Many of these E. coli strains carry P-fimbriae and produce alpha-haemolysin and a necrotizing cytotoxin (CNF1). Some of the frequently isolated types of extra-intestinal pathogenic E. coli from dogs, cats and humans were found to be highly genetically related but showed differences in their P-fimbrial adhesins which determine host specificity. Transmission of extra-intestinal and enteral pathogenic E. coli between dogs and humans was reported. Further research is needed, however, to determine the role of dogs and cats as transmission vectors of pathogenic E. coli strains to other animals and humans. PMID:10367359

Beutin, L

1999-01-01

293

Brief heat treatment causes a structural change and enhances cytotoxicity of the Escherichia coli ?-hemolysin.  

PubMed

?-Hemolysin (HLY) is an important virulence factor for uropathogenic Escherichia coli. HLY is a member of the RTX family of exotoxins secreted by a number of Gram-negative bacteria. Recently, it was reported that a related RTX toxin, the Mannheimia haemolytica leukotoxin, exhibits increased cytotoxicity following brief heat treatment. In this article, we show that brief heat treatment (1?min at 100°C) increases cytotoxicity of HLY for human bladder cells, kidney epithelial cells (A498) and neutrophils. Heat treatment also increased hemolysis of human red blood cells (RBCs). Furthermore, heat treatment of previously inactived HLY restored its cytotoxicity. Heat-activated and native HLY both required glycophorin A to lyse RBCs. Native and heat-activated HLY appeared to bind equally well to the surface of A498 cells; although, Western blot analyses demonstrated binding to different proteins on the surface. Confocal microscopy revealed that heat-activated HLY bound more extensively to internal structures of permeabilized A498 cells than did native HLY. Several lines of spectroscopic evidence demonstrate irreversible changes in the structure of heat activated compared to native HLY. We show changes in secondary structure, increased exposure of tryptophan residues to the aqueous environment, an increase in molecular dimension and an increase in hydrophobic surface area. These properties are among the most common characteristics described for the molten globule state, first identified as an intermediate in protein folding. We hypothesize that brief heat treatment of HLY causes a conformational change leading to significant differences in protein-protein interactions that result in increased cytotoxicity for target cells. PMID:22994841

Aulik, Nicole A; Atapattu, Dhammika N; Czuprynski, Charles J; McCaslin, Darrel R

2013-02-01

294

Fluorogenic assays for immediate confirmation of Escherichia coli.  

PubMed

Rapid assays for Escherichia coli were developed by using the compound 4-methylumbelliferone glucuronide (MUG), which is hydrolyzed by glucuronidase to yield a fluorogenic product. The production of glucuronidase was limited to strains of E. coli and some Salmonella and Shigella strains in the family Enterobacteriaceae. For immediate confirmation of the presence of E. coli in most-probable-number tubes, MUG was incorporated into lauryl tryptose broth at a final concentration of 100 micrograms/ml. Results of both the presumptive test (gas production) and the confirmed test (fluorescence) for E. coli were obtained from a variety of food, water, and milk samples after incubation for only 24 h at 35 degrees C. Approximately 90% of the tubes showing both gas production and fluorescence contained fecal coliforms (they were positive in EC broth incubated at 45 degrees C). Few false-positive reactions were observed. The lauryl tryptose broth-MUG-most-probable-number assay was superior to violet red bile agar for the detection of heat- and chlorine-injured E. coli cells. Anaerogenic strains produced positive reactions, and small numbers of E. coli could be detected in the presence of large numbers of competing bacteria. The fluorogenic assay was sensitive and rapid; the presence of one viable cell was detected within 20 h. E. coli colonies could be distinguished from other coliforms on membrane filters and plates of violet red bile agar if MUG was incorporated into the culture media. A rapid confirmatory test for E. coli that is amenable to automation was developed by using microtitration plates filled with a nonselective medium containing MUG. Pure or mixed cultures containing E. coli produced fluorescence within 4 h (most strains) to 24 h (a few weakly positive strains). PMID:7049088

Feng, P C; Hartman, P A

1982-06-01

295

Survival of Escherichia coli on strawberries grown under greenhouse conditions.  

PubMed

Strawberries are soft fruit that are not recommended to have a post-harvest wash due to quality concerns. Escherichia coli O157:H7 has been linked to outbreaks with strawberries but little is known about the survival of E. coli during the growth cycle of strawberries. The survival of E. coli on strawberry plants during growing under greenhouses conditions was evaluated. Soil, leaves, and strawberries (if present) were artificially contaminated with an E. coli surrogate either at the time of planting, first runner removal (4 wk), second runner removal (8 wk), or one week prior to harvest. At harvest E. coli was recovered from the leaves, soil, and strawberries regardless of the contamination time. Time of contamination influenced (P < 0.05) numbers of viable E. coli on the plant. The highest survival of E. coli (P < 0.0001) was detected in soil that was contaminated at planting (4.27 log10 CFU g soil(-1)), whereas, the survival of E. coli was maximal at later contamination times (8 wk and 1 wk prior to harvest) for the leaves (4.40 and 4.68 log10 CFU g leaves(-1)) and strawberries (3.37 and 3.53 log10 CFU strawberry(-1)). Cross contamination from leaves to fruit was observed during this study, with the presence of E. coli on strawberries which had not been present at the time of contamination. These results indicate that good agricultural best practices to avoid contamination are necessary to minimize the risk of contamination of these popular fruit with enteric pathogens. Practices should include soil testing prior to harvest and avoiding contamination of the leaves. PMID:25475285

Shaw, Angela Laury; Svoboda, Amanda; Jie, Beatrice; Nonnecke, Gail; Mendonca, Aubrey

2015-04-01

296

Expression of the Serratia marcescens lipoproteins gene in Escherichia coli.  

PubMed Central

The lipoprotein gene (lpp) of Serratia marcescens was cloned in a lambda phage vector (K. Nakamura and M. Inouye, Proc. Natl. Acad. Sci. U.S.A. 77: 1369-1373, 1980). This lpp gene was recloned in plasmid vectors pBR322 and pSC101. When a lipoprotein-deficient (lpp) mutant of Escherichia coli was transformed with pBR322 carrying the S. marcescens lpp gene, cells became nonleaky for ribonuclease, resistant to ethylenediaminetetraacetic acid, and sensitive to globomycin. The lipoprotein was found exclusively in the outer membrane fraction. These results indicate that the S. marcescens lipoprotein was normally secreted across the cytoplasmic membrane, modified, and assembled in the E. coli outer membrane. The amount of the free-form lipoprotein produced in this system was three times higher than that produced in lpp + C. coli cells, whereas there was no difference in the amount of the bound-form lipoprotein. On the other hand, lpp E. coli cells which harbored pSC101 carrying the S. marcescens lpp gene produced only one-third of the free-form lipoprotein produced in lpp E. coli cells which harbored pSC101 carrying the E. coli lpp gene. One of the major factors causing this difference in efficiency of gene expression between the lpp genes of S. marcescens and E. coli appears to be a deletion mutation at the transcription termination region found in the cloned S. marcescens lpp gene. The functional half-life of the S. marcescens lpp messenger ribonucleic acid in E. coli was found to be found half that of the E. coli lpp messenger ribonucleic acid. Images PMID:7016834

Lee, N; Nakamura, K; Inouye, M

1981-01-01

297

Bacteriophage cocktail significantly reduces Escherichia coli O157  

PubMed Central

Foods contaminated with Escherichia coli O157:H7 cause more than 63,000 foodborne illnesses in the United States every year, resulting in a significant economic impact on medical costs and product liabilities. Efforts to reduce contamination with E. coli O157:H7 have largely focused on washing, application of various antibacterial chemicals, and gamma-irradiation, each of which has practical and environmental drawbacks. A relatively recent, environmentally-friendly approach proposed for eliminating or significantly reducing E. coli O157:H7 contamination of foods is the use of lytic bacteriophages as biocontrol agents. We found that EcoShield™, a commercially available preparation composed of three lytic bacteriophages specific for E. coli O157:H7, significantly (p < 0.05) reduced the levels of the bacterium in experimentally contaminated beef by ? 94% and in lettuce by 87% after a five minute contact time. The reduced levels of bacteria were maintained for at least one week at refrigerated temperatures. However, the one-time application of EcoShield™ did not protect the foods from recontamination with E. coli O157:H7. Our results demonstrate that EcoShield™ is effective in significantly reducing contamination of beef and lettuce with E. coli O157:H7, but does not protect against potential later contamination due to, for example, unsanitary handling of the foods post processing. PMID:23275869

Carter, Chandi D.; Parks, Adam; Abuladze, Tamar; Li, Manrong; Woolston, Joelle; Magnone, Joshua; Senecal, Andre; Kropinski, Andrew M.; Sulakvelidze, Alexander

2012-01-01

298

Escherichia coli sequence type 131: epidemiology and challenges in treatment.  

PubMed

Escherichia coli ST131 has emerged as a global epidemic, multidrug-resistant clone of E. coli causing extra-intestinal infections. It is now highly prevalent among fluoroquinolone-resistant and CTX-M ESBL-producing E. coli isolates worldwide. Humans are likely the primary reservoir of ST131. Factors associated with its acquisition include residence in long-term care facilities and recent receipt of antimicrobial agents. E. coli ST131 causes a wide array of infections ranging from cystitis to life-threatening sepsis. Fluoroquinolones and trimethoprim-sulfamethoxazole are no longer adequate options for empiric therapy when E. coli ST131 is suspected from risk factors and local epidemiology. Expanded-spectrum cephalosporins, piperacillin-tazobactam and carbapenems are options to treat serious non-ESBL-producing E. coli ST131 infections, while carbapenems are indicated for ESBL-producing infections. There is a growing interest in reevaluating oral agents including fosfomycin and pivmecillinam for less serious infections such as uncomplicated cystitis. PMID:24694052

Qureshi, Zubair A; Doi, Yohei

2014-05-01

299

A miniaturized biotyping system for strain discrimination in Escherichia coli.  

PubMed

A two-tier miniaturized scheme of eight tests was devised for biotyping strains of Escherichia coli in microwell plates. Primary biotypes were defined by positive and negative reactions in tests for fermentation of raffinose, sorbose, dulcitol and 2-deoxy-D-ribose and for decarboxylation of ornithine when read after specified periods of incubation; subtypes were identified within primary biotypes according to results in secondary tests for rhamnose fermentation, lysine decarboxylation and motility. The method gave reproducible results on different occasions of testing. Among 100 E. coli strains from various sources, 26 of the 32 possible primary biotypes and 56 full biotypes, as defined by results in both primary and secondary tests, were identified, thus demonstrating a high index of strain discrimination (D = 0.98). The scheme is recommended as a simple, reliable, inexpensive and efficient method of differentiating strains of E. coli. PMID:8348935

Crichton, P B; Logan, J M; Old, D C

1993-08-01

300

Bronchopneumonia associated with extraintestinal pathogenic Escherichia coli in a horse.  

PubMed

Extraintestinal pathogenic Escherichia coli (ExPEC) strains carrying distinct virulence attributes are known to cause diseases in humans and animals and infect organs other than the gastrointestinal tract. A fatal case of bronchopneumonia in a 12-year-old female Quarterhorse was investigated. Following postmortem examination, E. coli, Enterococcus sp., and Klebsiella pneumonia were isolated from the lungs, which contained multifocal intra-alveolar accumulations of neutrophils and macrophages with edema, hemorrhage, and fibrin. The strain of E. coli belonged to O2H21 and carried virulence genes cnf1, sfa, foc, fimA, and papG allele I that are known to be associated with ExPEC strains. The strain was resistant to several antimicrobials including clindamycin, erythromycin, oxacillin, penicillin, and rifampin. This is the first report, to the authors' knowledge, in which ExPEC O2H21 has been associated with fatal bronchopneumonia in a horse. PMID:18776106

DebRoy, Chitrita; Roberts, Elisabeth; Jayarao, Bhushan M; Brooks, Jason W

2008-09-01

301

Engineering Escherichia coli K12 MG1655 to use starch  

PubMed Central

Background To attain a sustainable bioeconomy, fuel, or valuable product, production must use biomass as substrate. Starch is one of the most abundant biomass resources and is present as waste or as a food and agroindustry by-product. Unfortunately, Escherichia coli, one of the most widely used microorganisms in biotechnological processes, cannot use starch as a carbon source. Results We engineered an E. coli strain capable of using starch as a substrate. The genetic design employed the native capability of the bacterium to use maltodextrins as a carbon source plus expression and secretion of its endogenous ?-amylase, AmyA, in an adapted background. Biomass production improved using 35% dissolved oxygen and pH 7.2 in a controlled bioreactor. Conclusion The engineered E. coli strain can use starch from the milieu and open the possibility of optimize the process to use agroindustrial wastes to produce biofuels and other valuable chemicals. PMID:24886307

2014-01-01

302

Functions of the gene products of Escherichia coli.  

PubMed Central

A list of currently identified gene products of Escherichia coli is given, together with a bibliography that provides pointers to the literature on each gene product. A scheme to categorize cellular functions is used to classify the gene products of E. coli so far identified. A count shows that the numbers of genes concerned with small-molecule metabolism are on the same order as the numbers concerned with macromolecule biosynthesis and degradation. One large category is the category of tRNAs and their synthetases. Another is the category of transport elements. The categories of cell structure and cellular processes other than metabolism are smaller. Other subjects discussed are the occurrence in the E. coli genome of redundant pairs and groups of genes of identical or closely similar function, as well as variation in the degree of density of genetic information in different parts of the genome. PMID:7508076

Riley, M

1993-01-01

303

Effect of tannins on the in viro growth of Escherichia coli O157:H7 and in vivo growth of generic Escherichia coli excreted from steers  

Technology Transfer Automated Retrieval System (TEKTRAN)

The effect of commercially available chestnut and mimosa tannins in vitro (experiment 1) or in vivo (experiment 2) on the growth or recovery of Escherichia coli O157:H7 or generic fecal E. coli was evaluated. In experiment 1, the mean growth rate of E. coli O157:H7, determined via the measurement o...

304

Prevalence and virulence gene profiles of Shiga toxin-producing Escherichia coli and enteropathogenic Escherichia coli from diarrhoeic and healthy lambs in India  

Microsoft Academic Search

This communication describes the prevalence and virulence attributes of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) isolates in lambs with and without diarrhoea in Kashmir, India. One hundred twenty and 164 E. coli isolates belonging to 56 different ‘O’ serogroups were isolated from 101 diarrhoeic and 135 healthy lambs, respectively. All the 284 isolates were screened for

M. A. Bhat; Y. Nishikawa; S. A. Wani

2008-01-01

305

The influence of antibiotic resistance gene carriage on biofilm formation by two Escherichia coli strains associated with urinary tract infections.  

PubMed

Urinary tract infections (UTI) caused by uropathogenic Escherichia coli are one of the most common forms of human disease. In this study, the effect of the presence of newly acquired antibiotic resistance genes on biofilm formation of UTI-associated E. coli strains was examined. Two clinical UTI-associated E. coli strains (SMC18 and SMC20) carrying different combinations of virulence genes were transformed with pGEM-T, pGEM-T::Km?Amp, or pGEM-T::Km to construct ampicillin-resistant (Km(S)Amp(R)), kanamycin-resistant (Km(R)Amp(S)), or ampicillin- and kanamycin-resistant (Km(R)Amp(R)) strains. Transformed and wild-type strains were characterized for biofilm formation, bacterial surface hydrophobicity, auto-aggregation, morphology, and attachment to abiotic surfaces. Transformation with a plasmid carrying an ampicillin resistance gene alone decreased (p < 0.05) biofilm formation by SMC18 (8 virulence marker genes) but increased (p < 0.05) biofilm formation by SMC20 (5 virulence marker genes). On the other hand, transformation with a plasmid carrying a kanamycin resistance gene alone or both ampicillin and kanamycin resistance genes resulted in a decrease (p < 0.05) in biofilm formation by SMC18 but did not affect (p > 0.05) the biofilm formation by SMC20. Our results suggest that transformation of UTI-associated E. coli with plasmids carrying different antibiotic resistance gene(s) had a significant impact on biofilm formation and that these effects were both strain dependent and varied between different antibiotics. PMID:24498987

Teh, Amy Huei Teen; Wang, Yi; Dykes, Gary A

2014-02-01

306

EXPERIMENTAL MASTITIS WITH ESCHERICHIA COLI: SEQUENTIAL RESPONSE OF LEUKOCYTES AND OPSONIC ACTIVITY  

E-print Network

EXPERIMENTAL MASTITIS WITH ESCHERICHIA COLI: SEQUENTIAL RESPONSE OF LEUKOCYTES AND OPSONIC ACTIVITY infusion of bacteria (Hill, 1979); b) opsonic activity is required in milk to enable polymorphonuclear

Paris-Sud XI, Université de

307

Analysis of O-island deletions in Escherichia coli O157:H7   

E-print Network

Escherichia coli (E. coli) are a diverse species of bacteria that reside, often harmoniously and beneficially, in the gastrointestinal tracts of humans and other mammals. However, some strains are associated with serious ...

Flockhart, Allen Forrest

2012-11-30

308

Global dissemination of a multidrug resistant Escherichia coli clone.  

PubMed

Escherichia coli sequence type 131 (ST131) is a globally disseminated, multidrug resistant (MDR) clone responsible for a high proportion of urinary tract and bloodstream infections. The rapid emergence and successful spread of E. coli ST131 is strongly associated with several factors, including resistance to fluoroquinolones, high virulence gene content, the possession of the type 1 fimbriae FimH30 allele, and the production of the CTX-M-15 extended spectrum ?-lactamase (ESBL). Here, we used genome sequencing to examine the molecular epidemiology of a collection of E. coli ST131 strains isolated from six distinct geographical locations across the world spanning 2000-2011. The global phylogeny of E. coli ST131, determined from whole-genome sequence data, revealed a single lineage of E. coli ST131 distinct from other extraintestinal E. coli strains within the B2 phylogroup. Three closely related E. coli ST131 sublineages were identified, with little association to geographic origin. The majority of single-nucleotide variants associated with each of the sublineages were due to recombination in regions adjacent to mobile genetic elements (MGEs). The most prevalent sublineage of ST131 strains was characterized by fluoroquinolone resistance, and a distinct virulence factor and MGE profile. Four different variants of the CTX-M ESBL-resistance gene were identified in our ST131 strains, with acquisition of CTX-M-15 representing a defining feature of a discrete but geographically dispersed ST131 sublineage. This study confirms the global dispersal of a single E. coli ST131 clone and demonstrates the role of MGEs and recombination in the evolution of this important MDR pathogen. PMID:24706808

Petty, Nicola K; Ben Zakour, Nouri L; Stanton-Cook, Mitchell; Skippington, Elizabeth; Totsika, Makrina; Forde, Brian M; Phan, Minh-Duy; Gomes Moriel, Danilo; Peters, Kate M; Davies, Mark; Rogers, Benjamin A; Dougan, Gordon; Rodriguez-Baño, Jesús; Pascual, Alvaro; Pitout, Johann D D; Upton, Mathew; Paterson, David L; Walsh, Timothy R; Schembri, Mark A; Beatson, Scott A

2014-04-15

309

The metabolism of isocytidine in Escherichia coli  

PubMed Central

Intact cells and cell-free extracts of E. coli convert isocytidine to isocytosine and uracil. The radioactive label of 5-[3H]isocytidine is incorporated into RNA and, DNA of growing bacteria at a rate equal to about 1.4% of that of cytidine under similar conditions; the radioactivity is found in uridylic, cytidylic and 2?-deoxythymidylic acids, while less than 0.4% of incorporated radioactive material might be due to possible incorporation of intact isocytidine. Uridine phosphorylase and cytidine deaminase apparently do not participate in the metabolic conversion of isocytidine. 5-[3H]isocytidine was prepared by platinum-catalyzed dehalogenation of 5-bromoisocytidine in the presence of tritium. The 5-bromo derivative was obtained from 2?,3?-0- -isopropylideneisocytidine by N-bromsuccinimide bromination followed by acidic hydrolysis. PMID:10793683

Dosko?il, J.; Holý, A.; Filip, J.

1974-01-01

310

Predicting non-coding RNA genes in Escherichia coli with boosted genetic programming  

E-print Network

Several methods exist for predicting non-coding RNA (ncRNA) genes in Escherichia coli (E.coli therefore speculate that E.coli contains more ncRNA genes than previously estimated. INTRODUCTION Non. We describe a method that uses automatic discovery of sequence patterns to predict ncRNA genes in E.coli

Fernandez, Thomas

311

The distribution and genetic structure of Escherichia coli in Australian vertebrates: host and geographic effects  

Microsoft Academic Search

Escherichia coli was isolated from more than 2300 non-domesticated vertebrate hosts living in Australia. E. coli was most prevalent in mammals, less prevalent in birds and uncommon in fish, frogs and reptiles. Mammals were unlikely to harbour E. coli if they lived in regions with a desert climate and less likely to have E. coli if they lived in the

David M. Gordon; Ann Cowling

2003-01-01

312

Enteropathogenic Escherichia coli translocated intimin receptor, Tir, requires a specic chaperone for stable  

E-print Network

. Summary Enteropathogenic Escherichia coli (EPEC) secretes several Esps (E. coli-secreted proteins, 1992). Related pathogens that cause disease using similar mechanisms include enterohaemorrhagic E. coli (EHEC) O157:H7 (Francis et al., 1986; Tzipori et al., 1987, 1988), rabbit enteropathogenic E. coli

Strynadka, Natalie

313

Persistence of cellulitis-associated Escherichia coli DNA ngerprints in successive broiler  

E-print Network

is primarily caused by Escherichia coli. Previous research found that the E. coli isolates of cellulitis origin were unique to each ranch, suggesting that these E. coli were endemic within the ranch environment. To test the hypothesis that the E. coli associated with cellulitis are endemic in the litter

Singer, Randall

314

Microbial uropathogens and their antibiotic resistance profile from hospitalized patients in Central Alabama.  

PubMed

Urinary tract infections remain a common problem in inpatient care. They are highly challenging to provide effective initial therapy without sensitivity data. The purpose of this study was to survey the uropathogens and their sensitivity profile at a hospital in Central Alabama and to guide experiential antibiotic selection. This was the first reported study on bacterial uropathogens and their antibiotic resistance profile at this Central Alabama hospital. The survey period was between July 2009 and June 2010, a total of 473 urine cultures were reviewed and susceptibility testing was determined using the Clinical and Laboratory Standards Institute (CLSI) microdilution method. The results indicated that Escherichia coli (45.5%) was the most common organism, followed by Klebsiella pneumoniae (18.2%), Pseudomonas aeruginosa (10.1%), Proteus mirabilis (7.8%), Enterobacter cloacae (4.2%), methicillin-resistant Staphylococcus aureus (3.0%), Klebsiella oxytoca and Citrobacter freundii (1.5%), Morganella morganii (1.3%), and the other species (7.0%). For the 215 E. coli isolates, imipenem and cephalosporins (except for cefazolin) had the highest sensitivity (99-100%, P < 0.05). In contrast, ampicillin had the highest resistance (57%, P < 0.05) as compared to other antibiotics (about 30%) including ampicillin/ sulbactam, ciprofloxacin, levofloxacin, tetracycline, and trimethoprim/sulfamethoxazole. The major finding of this study was that ciprofloxacin, levofloxacin and trimethoprim/sulfamethoxazole had comparable sensitivity patterns for Escherichia coli, Klebsiella pneumoniae, Proteus mirabilis, and Enterobacter cloacae, the most common uropathogens at this Central Alabama hospital. Additionally, this study found that E. coli had a resistant rate of 31% to ciprofloxacin and levofloxacin compared to the resistance rate of 28.4% and 15.8% in earlier reports (Lee et al. 2010; Rattanaumpawan et al. 2010), likely indicating the continuing evolution of resistance due to antibiotic exposure. It is imperative to monitor the resistance of P. aeruginosa considering their high resistance to imipenem found in this study. PMID:23330509

Qian, Li; Camara, Tracy; Taylor, J Kyle; Jones, Kathy W

2012-01-01

315

The Pangenome Structure of Escherichia coli: Comparative Genomic Analysis of E. coli Commensal and Pathogenic Isolates? †  

PubMed Central

Whole-genome sequencing has been skewed toward bacterial pathogens as a consequence of the prioritization of medical and veterinary diseases. However, it is becoming clear that in order to accurately measure genetic variation within and between pathogenic groups, multiple isolates, as well as commensal species, must be sequenced. This study examined the pangenomic content of Escherichia coli. Six distinct E. coli pathovars can be distinguished using molecular or phenotypic markers, but only two of the six pathovars have been subjected to any genome sequencing previously. Thus, this report provides a seminal description of the genomic contents and unique features of three unsequenced pathovars, enterotoxigenic E. coli, enteropathogenic E. coli, and enteroaggregative E. coli. We also determined the first genome sequence of a human commensal E. coli isolate, E. coli HS, which will undoubtedly provide a new baseline from which workers can examine the evolution of pathogenic E. coli. Comparison of 17 E. coli genomes, 8 of which are new, resulted in identification of ?2,200 genes conserved in all isolates. We were also able to identify genes that were isolate and pathovar specific. Fewer pathovar-specific genes were identified than anticipated, suggesting that each isolate may have independently developed virulence capabilities. Pangenome calculations indicate that E. coli genomic diversity represents an open pangenome model containing a reservoir of more than 13,000 genes, many of which may be uncharacterized but important virulence factors. This comparative study of the species E. coli, while descriptive, should provide the basis for future functional work on this important group of pathogens. PMID:18676672

Rasko, David A.; Rosovitz, M. J.; Myers, Garry S. A.; Mongodin, Emmanuel F.; Fricke, W. Florian; Gajer, Pawel; Crabtree, Jonathan; Sebaihia, Mohammed; Thomson, Nicholas R.; Chaudhuri, Roy; Henderson, Ian R.; Sperandio, Vanessa; Ravel, Jacques

2008-01-01

316

Roles of Serine Accumulation and Catabolism in the Colonization of the Murine Urinary Tract by Escherichia coli CFT073?  

PubMed Central

A d-serine deaminase (DsdA) mutant of uropathogenic Escherichia coli strain CFT073 has a hypercolonization phenotype in a murine model of urinary tract infection (UTI) due to increased virulence gene expression by an unknown mechanism (B. J. Haugen et al., Infect. Immun. 75:278-289, 2007). DsdC is a d-serine-dependent activator of dsdXA transcription. DsdC may regulate the virulence genes responsible for hypercolonization. The loss of DsdA leads to increased intracellular accumulation of d-serine. In this study we show that deletion of the genes encoding l-serine deaminases SdaA and SdaB resulted in a mutant that accumulates higher intracellular levels of l-serine than CFT073. CFT073 sdaA sdaB has a mild competitive colonization defect whereas a CFT073 dsdA sdaA sdaB triple mutant shows a greater loss in competitive colonization ability. Thus, the inability to generate serine-specific catabolic products does not result in hypercolonization and the ability to catabolize serine represents a positive physiological trait during murine UTI. CFT073 dsdC and CFT073 dsdC dsdA mutants continue to outcompete the wild type in the UTI model. These results confirm that loss of DsdA activity results in the hypercolonization phenotype and that DsdC does not play a direct role in the elevated-colonization phenotype. Interestingly, a CFT073 dsdA mutant with deletions of d-serine transporter genes dsdX and cycA shows wild-type colonization levels of the bladder but is attenuated for kidney colonization. Thus, d-serine acts as a signal for hypercolonization and virulence gene expression by CFT073 dsdA, whereas overall catabolism of serine represents a positive Escherichia coli fitness trait during UTI. PMID:17785472

Anfora, Andrew T.; Haugen, Brian J.; Roesch, Paula; Redford, Peter; Welch, Rodney A.

2007-01-01

317

Recent Advances in Understanding Enteric Pathogenic Escherichia coli  

PubMed Central

SUMMARY Although Escherichia coli can be an innocuous resident of the gastrointestinal tract, it also has the pathogenic capacity to cause significant diarrheal and extraintestinal diseases. Pathogenic variants of E. coli (pathovars or pathotypes) cause much morbidity and mortality worldwide. Consequently, pathogenic E. coli is widely studied in humans, animals, food, and the environment. While there are many common features that these pathotypes employ to colonize the intestinal mucosa and cause disease, the course, onset, and complications vary significantly. Outbreaks are common in developed and developing countries, and they sometimes have fatal consequences. Many of these pathotypes are a major public health concern as they have low infectious doses and are transmitted through ubiquitous mediums, including food and water. The seriousness of pathogenic E. coli is exemplified by dedicated national and international surveillance programs that monitor and track outbreaks; unfortunately, this surveillance is often lacking in developing countries. While not all pathotypes carry the same public health profile, they all carry an enormous potential to cause disease and continue to present challenges to human health. This comprehensive review highlights recent advances in our understanding of the intestinal pathotypes of E. coli. PMID:24092857

Croxen, Matthew A.; Law, Robyn J.; Scholz, Roland; Keeney, Kristie M.; Wlodarska, Marta

2013-01-01

318

The Genetic Basis of Escherichia coli Pathoadaptation to Macrophages  

PubMed Central

Antagonistic interactions are likely important driving forces of the evolutionary process underlying bacterial genome complexity and diversity. We hypothesized that the ability of evolved bacteria to escape specific components of host innate immunity, such as phagocytosis and killing by macrophages (M?), is a critical trait relevant in the acquisition of bacterial virulence. Here, we used a combination of experimental evolution, phenotypic characterization, genome sequencing and mathematical modeling to address how fast, and through how many adaptive steps, a commensal Escherichia coli (E. coli) acquire this virulence trait. We show that when maintained in vitro under the selective pressure of host M? commensal E. coli can evolve, in less than 500 generations, virulent clones that escape phagocytosis and M? killing in vitro, while increasing their pathogenicity in vivo, as assessed in mice. This pathoadaptive process is driven by a mechanism involving the insertion of a single transposable element into the promoter region of the E. coli yrfF gene. Moreover, transposition of the IS186 element into the promoter of Lon gene, encoding an ATP-dependent serine protease, is likely to accelerate this pathoadaptive process. Competition between clones carrying distinct beneficial mutations dominates the dynamics of the pathoadaptive process, as suggested from a mathematical model, which reproduces the observed experimental dynamics of E. coli evolution towards virulence. In conclusion, we reveal a molecular mechanism explaining how a specific component of host innate immunity can modulate microbial evolution towards pathogenicity. PMID:24348252

Miskinyte, Migla; Sousa, Ana; Ramiro, Ricardo S.; de Sousa, Jorge A. Moura; Kotlinowski, Jerzy; Caramalho, Iris; Magalhães, Sara; Soares, Miguel P.; Gordo, Isabel

2013-01-01

319

The biotyping of Escherichia coli isolated from healthy farm animals.  

PubMed

A total of 2973 Escherichia coli, isolated from six different groups of animals, were examined for their ability to ferment adonitol, dulcitol, raffinose, rhamnose and sorbose in solid media. Twenty-nine fermentation patterns were recorded although 2443 (82%) of the E. coli belonged to seven of the 32 possible biotypes. Ninety-six O-serotypes were identified within the 2973 E. coli. The number of O-serotypes represented in the 15 most common biotypes ranged from three to 15. Serotypes O8 and O9 were found most commonly in the different groups of animals and several biotypes amongst these two O-serotypes were identified in two or more groups of the animals. The ability of the E. coli to metabolize aesculin, ornithine, salicin and sucrose was also assessed. These test proved less reproducible and were not included in the primary biotyping scheme although their use allowed the enumeration of additional biotypes. The application of biotyping to the study of the ecology of drug-resistant strains of E. coli in five situations is briefly presented. PMID:7045219

Hinton, M; Allen, V; Linton, A H

1982-06-01

320

Unusual “Flesh-Eating” Strains of Escherichia coli  

PubMed Central

Monomicrobial necrotizing fasciitis (type II) is typically caused by group A streptococcus alone or in combination with Staphylococcus aureus. Escherichia coli has been isolated from polymicrobial or Fournier's gangrene but has rarely been reported in monomicrobial necrotizing fasciitis. We describe the clinical characteristics and outcomes of seven cases of monomicrobial E. coli necrotizing fasciitis and/or severe soft tissue infection diagnosed at a single institution during an 18-month period. Four isolates from three patients and two isolates from two patients with type I polymicrobial severe soft tissue infection (controls) were assayed by the randomly amplified polymorphic DNA (RAPD) analysis for fingerprinting and PCR amplification of primers in order to detect cytotoxic necrotizing factor 1 and 2 (cnf1 and cnf2) genes. All patients had some type of immune suppression. The limb was the most commonly involved organ. In all cases, E. coli was isolated as a monomicrobial pathogen from blood, fascia, or both. All patients died during hospitalization, three within the first 48 h. The RAPD amplification assay showed a high degree of genetic diversity among the “flesh-eating” strains and controls. The cnf1 toxin gene was identified in two out of three cases, but not in the controls. cnf2 was not detected in any of the patients. E. coli may be responsible for life-threatening necrotizing fasciitis. Further research is needed to reveal relevant risk factors, reservoirs, and modes of transmission of cnf1 E. coli. PMID:23035196

Shaked, Hila; Samra, Zmira; Paul, Michal; Madar-Shapiro, Liora; Cohen, Jonathan; Pitlik, Silvio

2012-01-01

321

CNF producing Escherichia coli isolated from cattle in Northern Ireland.  

PubMed

Tissue culture assays were used to investigate the incidence of cytotoxic necrotising factors (CNFs) 1 and 2 in Escherichia coli strains from cattle. E. coli cultures were obtained from faeces collected from 223 cases of diarrhoea and from 113 healthy animals. In addition, strains cultured from 62 cases of mastitis, 66 cases of septicaemia and 68 cases of abortion were also investigated. E. coli producing CNF 1 or 2 were identified in all sample groups except for the abortion cases. Comparable levels of CNF1 strains were present in E. coli from the faces of diarrhoeic (4%) and healthy faeces (4.4%) whereas lower levels of CNF2 were identified in the faeces from diarrhoeic animals (19.3%) in comparison with healthy animals (30.9%). One CNF1 producing strain was identified among the E. coli isolated from mastitis samples, while 3% and 10.6% of septicaemic strains were positive for CNF1 and 2, respectively. Serogrouping of CNF isolates did not reveal the association of any particular serogroups with the different conditions. PMID:8734640

Burns, A L; Ball, H J; Finlay, D A

1996-04-01

322

Unusual "flesh-eating" strains of Escherichia coli.  

PubMed

Monomicrobial necrotizing fasciitis (type II) is typically caused by group A streptococcus alone or in combination with Staphylococcus aureus. Escherichia coli has been isolated from polymicrobial or Fournier's gangrene but has rarely been reported in monomicrobial necrotizing fasciitis. We describe the clinical characteristics and outcomes of seven cases of monomicrobial E. coli necrotizing fasciitis and/or severe soft tissue infection diagnosed at a single institution during an 18-month period. Four isolates from three patients and two isolates from two patients with type I polymicrobial severe soft tissue infection (controls) were assayed by the randomly amplified polymorphic DNA (RAPD) analysis for fingerprinting and PCR amplification of primers in order to detect cytotoxic necrotizing factor 1 and 2 (cnf1 and cnf2) genes. All patients had some type of immune suppression. The limb was the most commonly involved organ. In all cases, E. coli was isolated as a monomicrobial pathogen from blood, fascia, or both. All patients died during hospitalization, three within the first 48 h. The RAPD amplification assay showed a high degree of genetic diversity among the "flesh-eating" strains and controls. The cnf1 toxin gene was identified in two out of three cases, but not in the controls. cnf2 was not detected in any of the patients. E. coli may be responsible for life-threatening necrotizing fasciitis. Further research is needed to reveal relevant risk factors, reservoirs, and modes of transmission of cnf1 E. coli. PMID:23035196

Shaked, Hila; Samra, Zmira; Paul, Michal; Madar-Shapiro, Liora; Cohen, Jonathan; Pitlik, Silvio; Bishara, Jihad

2012-12-01

323

Environmental Factors Affecting Indole Production in Escherichia coli  

PubMed Central

A variety of both Gram-positive and Gram-negative bacteria produce large quantities of indole as an intracellular signal in microbial communities. Biosynthesis of indole is well-studied, and while carbon sources and amino acids are important environmental cues for indole production in Escherichia coli, other environmental factors affecting indole production for this strain are less clear. This study demonstrates that the environmental cue pH is an important factor for indole production that further controls biofilm formation of E. coli. Moreover, E. coli produced a higher level of extracellular indole in the presence of the antibiotics ampicillin and kanamycin, and the increased indole enhanced cell survival during antibiotic stress. Additionally, we found here that temperature is another important factor for indole production; E. coli produces and accumulates a large amount of indole at 50°C, even at low cell densities. Overall, our results suggest that indole is a stable biological compound, and E. coli may utilize indole to protect itself against other microorganisms. PMID:21145393

Han, Thi Hiep; Lee, Jin-Hyung; Cho, Moo Hwan; Wood, Thomas K.; Lee, Jintae

2011-01-01

324

Immune complex glomerulonephritis in mice infected with Escherichia coli.  

PubMed Central

C57Bl/6 mice were injected intraperitoneally with 10(8) to 2 x 10(8) living K 38 Escherichia coli (E. coli) and serological changes and kidney involvement were studied. E. coli were found in the blood 45 min to 24 hr after injection. In serum, large amounts of deoxyribonucleic acid (DNA) were present 24 hr after E. coli injection, and thereafter disappeared. Seven days after infection, antibodies directed against E. coli, anti-DNA antibodies and C1q-binding substances were found in serum and the kinetics of the variations of these parameters were studied until day 35. Kidney lesions were evaluated immunochemically and by optical and electron microscopy. In the glomeruli, heavy granular deposits of IgG and IgM were constantly found in mesangium and along capillary walls. In most kidneys slight granular deposits of IgG and IgM were also found in the tubules. Histological studies revealed in the glomeruli mild endocapillary cell proliferation, focal thickening of glomerular basement membrane and dense deposits in mesangial and subendothelial areas and inside the glomerular basement membrane; in the tubules dense deposits were focally observed inside the tubular basement membrane. Images Fig. 6 Fig. 7 PMID:6450654

Fournié, G J; Mignon-Conté, M A; Lulé, J; Gayral-Ta Minh, M; Haas, S; Bauriaud, R; Conté, J J

1980-01-01

325

Isolation of Enteropathogenic Escherichia coli from lettuce samples in Tehran  

PubMed Central

Aim The purpose of this study was to find the isolation rate of enteropathogenic Escherichia coli (EPEC) from lettuce samples collected in Tehran. Background During the last decade, the prevalence of infectious diarrheal diseases due to consumption of contaminated food especially raw vegetable has been increasingly reported. Enteropathogenic Escherichia coli strains are an important group of diarrheagenic E. coli that can cause infant diarrhea especially in the developing world. Material and Methods One hundred lettuce samples collected in Tehran were transported to the laboratory, homogenized by a stomacher in EC broth containing cefixime, and cultured on MacConkey agar plates. Bacterial DNA was extracted by boiling method and PCR was performed using three pairs of primers targeting stx1, stx2 and eaeA genes. Results Screening of 100 lettuce samples by PCR showed four samples were positive for the presence of EPEC. Conclusion This study suggests contamination of the lettuce by the EPEC and its possible role as the source of infection in this region. PMID:25436096

Mazaheri, Somayeh; Salmanzadeh-Ahrabi, Siavosh; Aslani, Mohammad-Mehdi

2014-01-01

326

Widespread antibiotic resistance of diarrheagenic Escherichia coli and Shigella species  

PubMed Central

Background: Antibiotic resistance of enteric pathogens particularly Shigella species, is a critical world-wide problem and monitoring their resistant pattern is essential, because the choice of antibiotics is absolutely dependent on regional antibiotic susceptibility patterns. During summer 2013, an unusual increase in number of diarrheal diseases was noticed in Isfahan, a central province of Iran. Therefore, the antibiotic resistance of diarrheagenic Escherichia coli and Shigella species isolated were evaluated. Materials and Methods: According to the guideline on National Surveillance System for Foodborn Diseases, random samples from patients with acute diarrhea were examined in local laboratories of health centers and samples suspicious of Shigella spp. were further assessed in referral laboratory. Isolated pathogens were identified by standard biochemical and serologic tests and antibiotic susceptibility testing was carried out by disc diffusion method. Results: A total of 1086 specimens were obtained and 58 samples suspicious of Shigella were specifically evaluated. The most prevalent isolated pathogen was Shigella sonnei (26/58) followed by E. coli (25/58) and Shigella flexneri (3/58). A large number of isolated bacteria were resistant to co-trimoxazole (Shigella spp: 100%, E. coli: 80%), azithromycin (Shigella spp: 70.4%, E. coli: 44.0%), ceftriaxone (Shigella spp: 88.9%, E. coli: 56.0%) and cefixime (Shigella spp: 85.2%, E. coli: 68.0%). About88.3% of S. sonnei isolates, one S. flexneri isolate, and 56% of E. coli strains were resistant to at least three antibiotic classes (multidrug resistant). Conclusion: Due to high levels of resistance to recommended and commonly used antibiotics for diarrhea, continuous monitoring of antibiotic resistance seems essential for determining best options of empirical therapy. PMID:25002896

Sadeghabadi, Azam Fatahi; Ajami, Ali; Fadaei, Reza; Zandieh, Masoud; Heidari, Elham; Sadeghi, Mahmoud; Ataei, Behrooz; Hoseini, Shervin Ghaffari

2014-01-01

327

Glucosylation of Isoflavonoids in Engineered Escherichia coli  

PubMed Central

A glycosyltransferase, YjiC, from Bacillus licheniformis has been used for the modification of the commercially available isoflavonoids genistein, daidzein, biochanin A and formononetin. The in vitro glycosylation reaction, using UDP-?-D-glucose as a donor for the glucose moiety and aforementioned four acceptor molecules, showed the prominent glycosylation at 4? and 7 hydroxyl groups, but not at the 5th hydroxyl group of the A-ring, resulting in the production of genistein 4?-O-?-D-glucoside, genistein 7-O-?-D-glucoside (genistin), genistein 4?,7-O-?-D-diglucoside, biochanin A-7-O-?-D-glucoside (sissotrin), daidzein 4?-O-?-D-glucoside, daidzein 7-O-?-D-glucoside (daidzin), daidzein 4?, 7-O-?-D-diglucoside, and formononetin 7-O-?-D-glucoside (ononin). The structures of all the products were elucidated using high performance liquid chromatography-photo diode array and high resolution quadrupole time-of-flight electrospray ionization mass spectrometry (HR QTOFESI/MS) analysis, and were compared with commercially available standard compounds. Significantly higher bioconversion rates of all four isoflavonoids was observed in both in vitro as well as in vivo bioconversion reactions. The in vivo fermentation of the isoflavonoids by applying engineered E. coli BL21(DE3)/?pgi?zwf?ushA overexpressing phosphoglucomutase (pgm) and glucose 1-phosphate uridyltransferase (galU), along with YjiC, found more than 60% average conversion of 200 ?M of supplemented isoflavonoids, without any additional UDP-?-D-glucose added in fermentation medium, which could be very beneficial to large scale industrial production of isoflavonoid glucosides. PMID:24599002

Pandey, Ramesh Prasad; Parajuli, Prakash; Koirala, Niranjan; Lee, Joo Ho; Park, Yong Il; Sohng, Jae Kyung

2014-01-01

328

IraL Is an RssB Anti-adaptor That Stabilizes RpoS during Logarithmic Phase Growth in Escherichia coli and Shigella  

PubMed Central

ABSTRACT RpoS (?S), the general stress response sigma factor, directs the expression of genes under a variety of stressful conditions. Control of the cellular ?S concentration is critical for appropriately scaled ?S-dependent gene expression. One way to maintain appropriate levels of ?S is to regulate its stability. Indeed, ?S degradation is catalyzed by the ClpXP protease and the recognition of ?S by ClpXP depends on the adaptor protein RssB. Three anti-adaptors (IraD, IraM, and IraP) exist in Escherichia coli K-12; each interacts with RssB and inhibits RssB activity under different stress conditions, thereby stabilizing ?S. Unlike K-12, some E. coli isolates, including uropathogenic E. coli strain CFT073, show comparable cellular levels of ?S during the logarithmic and stationary growth phases, suggesting that there are differences in the regulation of ?S levels among E. coli strains. Here, we describe IraL, an RssB anti-adaptor that stabilizes ?S during logarithmic phase growth in CFT073 and other E. coli and Shigella strains. By immunoblot analyses, we show that IraL affects the levels and stability of ?S during logarithmic phase growth. By computational and PCR-based analyses, we reveal that iraL is found in many E. coli pathotypes but not in laboratory-adapted strains. Finally, by bacterial two-hybrid and copurification analyses, we demonstrate that IraL interacts with RssB by a mechanism distinct from that used by other characterized anti-adaptors. We introduce a fourth RssB anti-adaptor found in E. coli species and suggest that differences in the regulation of ?S levels may contribute to host and niche specificity in pathogenic and nonpathogenic E. coli strains. PMID:24865554

Hryckowian, Andrew J.; Battesti, Aurelia; Lemke, Justin J.; Meyer, Zachary C.

2014-01-01

329

Inhibitory activity of cranberry juice on adherence of type 1 and type P fimbriated Escherichia coli to eucaryotic cells.  

PubMed Central

Inhibition of bacterial adherence to bladder cells has been assumed to account for the beneficial action ascribed to cranberry juice and cranberry juice cocktail in the prevention of urinary tract infections (A. E. Sobota, J. Urol. 131:1013-1016, 1984). We have examined the effect of the cocktail and juice on the adherence of Escherichia coli expressing surface lectins of defined sugar specificity to yeasts, tissue culture cells, erythrocytes, and mouse peritoneal macrophages. Cranberry juice cocktail inhibited the adherence of urinary isolates expressing type 1 fimbriae (mannose specific) and P fimbriae [specific for alpha-D-Gal(1----4)-beta-D-Gal] but had no effect on a diarrheal isolate expressing a CFA/I adhesin. The cocktail also inhibited yeast agglutination by purified type 1 fimbriae. The inhibitory activity for type 1 fimbriated E. coli was dialyzable and could be ascribed to the fructose present in the cocktail; this sugar was about 1/10 as active as methyl alpha-D-mannoside in inhibiting the adherence of type 1 fimbriated bacteria. The inhibitory activity for the P fimbriated bacteria was nondialyzable and was detected only after preincubation of the bacteria with the cocktail. Cranberry juice, orange juice, and pineapple juice also inhibited adherence of type 1 fimbriated E. coli, most likely because of their fructose content. However, the two latter juices did not inhibit the P fimbriated bacteria. We conclude that cranberry juice contains at least two inhibitors of lectin-mediated adherence of uropathogens to eucaryotic cells. Further studies are required to establish whether these inhibitors play a role in vivo. PMID:2653218

Zafriri, D; Ofek, I; Adar, R; Pocino, M; Sharon, N

1989-01-01

330

A second Escherichia coli protein with CL synthase activity.  

PubMed

The Escherichia coli open reading frame f413, which has the potential to code for a polypeptide homologous to cardiolipin (CL) synthase, has been cloned. Its polypeptide product has a molecular mass of 48 kDa, is membrane-bound, and catalyzes CL formation but does not hydrolyze CL. A comparison of the sequences predicted for the polypeptides encoded by f413 and cls indicates that the N-terminal residues specified by cls may be unnecessary for CL synthase activity. Construction of a truncated cls gene and characterization of its polypeptide product have confirmed this conclusion. PMID:10634942

Guo, D; Tropp, B E

2000-01-17

331

L Tyrosine production by deregulated strains of Escherichia coli  

Microsoft Academic Search

The excretion of the aromatic amino acid l-tyrosine was achieved by manipulating three gene targets in the wild-type Escherichia coli K12: The feedback-inhibition-resistant (fbr) derivatives of aroG and tyrA were expressed on a low-copy-number vector, and the TyrR-mediated regulation of the aromatic amino acid biosynthesis was\\u000a eliminated by deleting the tyrR gene. The generation of this l-tyrosine producer, strain T1,

Tina Lütke-Eversloh; Gregory Stephanopoulos

2007-01-01

332

[Characterization of Escherichia coli esterase B after separation by chromatography].  

PubMed

Esterase B of Escherichia coli has been purified 56 fold with recovery of 39%. The apparent molecular weight as determined by gel filtration was approximately 57000. The pI as determined by isoelectric focusing was 4.6. This enzyme exhibited Michaelis-Menton kinetics with apparent Km of 0.25 mM for l-naphtyl acetate. It remained stable at 60 degrees C but was sensitive to pH values below 6. The esterase activity was completely inhibited by Di-isopropyl-fluorophosphate (DFP) but was resistant to iodoacetamide and to EDTA. PMID:6405981

Goullet, P; Picard, B; Hieng, H S

1983-01-31

333

Urinary-tract infection: localisation and virulence of Escherichia coli.  

PubMed

Virulence of 15 strains of Escherichia coli from the human upper urinary tract was compared with that of 16 strains from the lower urinary tract, using an ascending infection in the mouse. No significant difference was found. There was no significant difference in frequency of K antigen and ability to ferment dulcitol between 32 lower strains and 31 upper strains. However, 22 strains containing K antigen, regardless of anatomical site of localisation, were more significantly likely to cause infection than 9 strains with no antigen. Similarly, 23 dulcitolfermenting strains, regardless of site of localisation, were significantly more likely to cause infection than 8 non-fermenting strains. PMID:46051

Kalmanson, G M; Harwick, H J; Turck, M; Guze, L B

1975-01-18

334

Mounting of Escherichia coli spheroplasts for AFM imaging.  

SciTech Connect

The cytoplasmic membrane of Escherichia coli (E. coli) is the location of numerous, chemically specific transporters and recognition elements. Investigation of this membrane in vivo by atomic force microscopy (AFM) requires removal of the cell wall and stable immobilization of the spheroplast. AFM images demonstrate that spheroplasts can be secured with warm gelatin applied to the mica substrate just before the addition of a spheroplast suspension. The resulting preparation can be repeatedly imaged by AFM over the course of several hours. Confocal fluorescence imaging confirms the association of the spheroplasts with the gelatin layer. Gelatin molecules are known to reorder into a network after heating. Entrapment within this gelatin network is believed to be responsible for the immobilization of spheroplasts on mica.

Sullivan, Claretta J [ORNL; Morrell-Falvey, Jennifer L [ORNL; Allison, David P [ORNL; Doktycz, Mitchel John [ORNL

2005-11-01

335

Impact of cranberry on Escherichia coli cellular surface characteristics  

SciTech Connect

The anti-adhesive effects of cranberry have been attributed to both interactions of its components with the surface of bacterial cells and to inhibition of p-fimbriae expression. Previous reports also suggested that the presence of cranberry juice changed the Gram stain characteristics of Escherichia coli. Here, we show that the morphology of E. coli is changed when grown in the presence of juice or extract from Vaccinium macrocarpon (cranberry). Gene expression analysis indicates the down regulation of flagellar basal body rod and motor proteins. Consistent with this finding and previous reports, the SEM images indicate a decrease in the visible p-fimbriae. The iodine used in Gram-staining protocols was found to interact differently with the bacterial membrane when cells were cultured in spiked media. Slight alterations in the Gram stain protocol demonstrated that culturing in the presence of cranberry juice does not change the Gram stain characteristics contradicting other reports.

Johnson, Brandy J. [Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Ave SW, Code 6900, Washington, DC 20375 (United States)], E-mail: brandy.white@nrl.navy.mil; Lin Baochuan; Dinderman, Michael A. [Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Ave SW, Code 6900, Washington, DC 20375 (United States); Rubin, Robert A. [Independent Researcher, 8620 Portafino Place, Whittier, CA 90603 (United States); Malanoski, Anthony P.; Ligler, Frances S. [Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 4555 Overlook Ave SW, Code 6900, Washington, DC 20375 (United States)

2008-12-19

336

Cellulosic hydrolysate toxicity and tolerance mechanisms in Escherichia coli  

PubMed Central

The sustainable production of biofuels will require the efficient utilization of lignocellulosic biomass. A key barrier involves the creation of growth-inhibitory compounds by chemical pretreatment steps, which ultimately reduce the efficiency of fermentative microbial biocatalysts. The primary toxins include organic acids, furan derivatives, and phenolic compounds. Weak acids enter the cell and dissociate, resulting in a drop in intracellular pH as well as various anion-specific effects on metabolism. Furan derivatives, dehydration products of hexose and pentose sugars, have been shown to hinder fermentative enzyme function. Phenolic compounds, formed from lignin, can disrupt membranes and are hypothesized to interfere with the function of intracellular hydrophobic targets. This review covers mechanisms of toxicity and tolerance for these compounds with a specific focus on the important industrial organism Escherichia coli. Recent efforts to engineer E. coli for improved tolerance to these toxins are also discussed. PMID:19832972

Mills, Tirzah Y; Sandoval, Nicholas R; Gill, Ryan T

2009-01-01

337

Involvement of DNA superhelicity in minichromosome maintenance in Escherichia coli.  

PubMed Central

Evidence is presented that Escherichia coli minichromosomes are harbored at superhelical densities which are lower than those measured for other E. coli plasmids but are comparable to that of the chromosome. When introduced into gyrB decreased-supercoiling mutants, minichromosomes were much more unstable than in strains with normal or increased supercoiling properties; in fact, certain minichromosome derivatives could not be introduced into top gyrB decreased-supercoiling mutants. These observations were unique to minichromosomes, since the maintenance of plasmids which did not replicate from oriC was not altered in these mutants. Analyses of minichromosomes of identical sizes but with different restriction fragment orientations suggested that supercoiling-dependent alterations in promoter-terminator functions, as well as direct effects of supercoiling on replication, may play a role in the observed minichromosome instability. Images PMID:2981821

Leonard, A C; Whitford, W G; Helmstetter, C E

1985-01-01

338

Continuous-sterilization system that uses photosemiconductor powders. [Escherichia coli  

SciTech Connect

We report a novel photochemical sterilization system in which Escherichia coli cells were sterilized with photosemiconductor powders (titanium oxide). For sterilization that could be used in practice, it was necessary to separate the TiO/sub 2/ powders from the cell suspension. Therefore, semiconductor powders were immobilized on acetylcellulose membranes. We constructed a continuous-sterilization system consisting of TiO/sub 2/-immobilized acetylcellulose membrane reactor, a mercury lamp, and a masterflex pump. As a result, under the various sterilization conditions examined, E.coli (10/sup 2/ cells per ml) was sterilized to < 1% survival when the cell suspension flowed in this system at a mean residence time of 16.0 min under irradiation (1800 microeinsteins/m/sup 2/ per s). We found that this system was reusable.

Matsunaga, T.; Tomoda, R.; Nakajima, T.; Nakamura, N.; Komine, T.

1988-06-01

339

Escherichia coli response to exogenous pyrophosphate and analogs.  

PubMed

The addition of exogenous pyrophosphate increases the growth yield and cAMP synthesis in stationary phase when Escherichia coli is grown in minimal medium. Pyrophosphate increases the yield by altering the enterobactin uptake system. We studied the physiological effects and examined how the E. coli transcriptome was modified when two structural analogs of pyrophosphate were added to the growth medium. Methylenediphosphonic acid or a high concentration of iron had the same positive effects as pyrophosphate on growth yield, cAMP synthesis and the repression of Fur-regulated genes. In contrast, imidodiphosphate did not affect these cellular processes significantly. The transcriptome modifications generated by pyrophosphate or methylenediphosphonic acid were more similar than those generated by imidodiphosphate or excess iron. The transcriptome data also indicated that processes other than iron uptake might be involved in the cellular response to exogenous pyrophosphate or methylenediphosphonic acid. PMID:12673060

Biville, Francis; Oshima, Taku; Mori, Hirotada; Kawagoe, Yuya; Bouvet, Odile; Rager, Marie-Noëlle; Perrotte-Piquemal, Marina; Danchin, Antoine

2003-01-01

340

Four products from Escherichia coli pseudogenes increase hydrogen production.  

PubMed

Pseudogenes are considered to be nonfunctional genes that lack a physiological role. By screening 3985 Escherichia coli mutants using chemochromic membranes, we found four pseudogenes involved in hydrogen metabolism. Knockouts of pseudogenes ydfW and ypdJ had a defective hydrogen phenotype on glucose and formate, respectively. Also, the knockout of pseudogene yqiG formed hydrogen from formate but not from glucose. For the yqiG mutant, 100% hydrogen recovery was obtained by the complementation of YqiG via a plasmid. The knockout of pseudogene ylcE showed hydrogen deficiency in minimal media which suggested that the role of YlcE is associated with cell growth. Hence, the products of these four pseudogenes play an important physiological role in hydrogen production in E. coli. PMID:24025676

Mohd Yusoff, Mohd Zulkhairi; Hashiguchi, Yuya; Maeda, Toshinari; Wood, Thomas K

2013-10-01

341

A biotyping scheme for the subspecific discrimination of Escherichia coli.  

PubMed

A two-tier system for biotyping Escherichia coli gave a fine and reliable differentiation of strains and is capable of future extension by the addition of new types and new tests. Strains were allocated to a primary biotype (1-16) by their reactions in four primary tests in culture media containing raffinose, sorbose, ornithine or dulcitol. Subtypes were distinguished within the primary biotypes by reactions in six secondary tests for rhamnose fermentation, lysine decarboxylation, aesculin hydrolysis, motility, type-1 fimbriation and prototrophy. Full biotypes were designated by letters indicating subtype reactions appended to primary type numbers. A series of 599 strains (1242 cultures) of E. coli from diverse sources was classified into 16 primary biotypes and into 213 full biotypes. The biotype characters of a strain were generally stable during its spread in the natural environment and in non-selective media used for storage. PMID:6754944

Crichton, P B; Old, D C

1982-05-01

342

A concise biotyping system for differentiating strains of Escherichia coli.  

PubMed

A six test biotyping system comprising fermentation of dulcitol, sorbose, raffinose, and 5 ketogluconate, motility and production of beta-haemolysis was used to obtain biotype profiles for 514 strains of Escherichia coli isolated from the urinary tract. This profile was suffixed to the API-20E code, recorded when the strains were originally identified. An expanded and reliable biotyping system was thus created giving a greater number of possible biotypes than with either system alone. The sensitivity in terms of distinguishing the organisms studied was therefore greatly increased. The use of O-serotyping in combination with biotyping is discussed. Biotyping can also be usefully supplemented by the determination of the eight commonest E coli O-serotypes. This is of value in many clinical situations where differentiation of organisms is vital to the proper analysis of results. PMID:6757274

Gargan, R; Brumfitt, W; Hamilton-Miller, J M

1982-12-01

343

Genetic characterization of moaB mutants of Escherichia coli  

PubMed Central

The moaABCDE operon of Escherichia coli encodes enzymes essential for the biosynthesis of the molybdenum cofactor (Moco). However, the role of the moaB gene within this operon has remained enigmatic. Here, we have investigated the effect of moaB defects on two phenotypes diagnostic for Moco-deficiency: chlorate-resistance and sensitivity to the base analog 6-N-hydroxylaminopurine (HAP). We found that transposon insertions in moaB caused partial Moco-deficiency associated with chlorate-resistance, but not for HAP-sensitivity. On the other hand, in-frame deletions of moaB, or moaB overexpression, had no effect on either phenotype. Our combined data are consistent with the lack of any role for MoaB in Moco biosynthesis in E. coli. PMID:23680484

Kozmin, Stanislav G.; Schaaper, Roel M.

2013-01-01

344

Enterotoxigenic and necrotizing Escherichia coli in human diarrhoea in Spain.  

PubMed

Enterotoxigenic Escherichia coli (ETEC) strains of serotype 0153: K-:H45 CFA/I+ STa+ were associated with two outbreaks of neonatal diarrhoea that occurred in two different hospitals of Madrid, in one of which several children died. Two other outbreaks were associated with ETEC strains of serotypes 0159: K-:H21 (LT+) and 0159: K-:H4 (LT+ STa+) without CFA/I and CFA/II colonization factors. Necrotizing E. coli (NTEC) strains of serotype 06:K13, producing the cytotoxic necrotizing factor CNF1 and alpha-haemolysin, were also associated with two outbreaks of neonatal diarrhoea that occurred in a hospital in Madrid and in a hospital in Talavera de la Reina. The results of the characterization of some ETEC and NTEC strains isolated from sporadic cases of diarrhoea are also discussed. PMID:1397224

Blanco, J; González, E A; Espinosa, P; Blanco, M; Garabal, J I; Alonso, M P

1992-07-01

345

Deer sausage: a newly identified vehicle of transmission of Escherichia coli O157:H7.  

PubMed

Five Missouri patients infected with Escherichia coli O157:H7 were studied for an epidemiologically plausible association. Case isolates, case interviews, and pathogen and meat XbaI pulsed field electrophoresis patterns were consistent with the common source being contaminated, fermented deer sausage, a previously unrecognized mode of transmission for Escherichia coli O157:H7. PMID:19773004

Ahn, Christina K; Russo, Anthony J; Howell, Karla R; Holt, Nicholas J; Sellenriek, Patricia L; Rothbaum, Robert J; Beck, Anne M; Luebbering, Leon J; Tarr, Phillip I

2009-10-01

346

FACTORS INFLUENCING THE SHEDDING OF ESCHERICHIA COLI AND SALMONELLA SPP. IN HOLSTEIN CATTLE  

Technology Transfer Automated Retrieval System (TEKTRAN)

Fecal samples were obtained in replicate from lactating (LAC; n = 60) and non-lactating (NLAC; n = 60) Holstein cows to determine the influence of time of day (AM vs PM), parity, and lactation phase [ 60 d in milk (DIM)] on shedding of Escherichia coli O157:H7 (EHEC), Escherichia coli (EC),...

347

Antibiotics Susceptibility Pattern of Escherichia coli Strains Isolated from Chickens with Colisepticemia in Tabriz Province, Iran  

Microsoft Academic Search

Antimicrobial agents are used extremely in order to reducing the enormous losses caused by Escherichia coli infections (colibacillosis) in Iran poultry industry. In this investigation fifty avian pathogenic Escherichia coli (APEC) strains isolated from broiler chickens with colisepticemia and examined for susceptibility to antimicrobials of veterinary and human significance. In vitro antibiotic activities of 3 2 antibiotic substances against the

2006-01-01

348

Structural and Biochemical Characterization of a Quinol Binding Site of Escherichia coli Nitrate Reductase A*  

E-print Network

Structural and Biochemical Characterization of a Quinol Binding Site of Escherichia coli Nitrate of Escherichia coli nitrate re- ductase A (NarGHI) in complex with pentachlorophenol has been determined to 2.0 Ã? of resolution. We have shown that pentachlorophenol is a potent inhibitor of quinol:nitrate oxidoreductase

Strynadka, Natalie

349

Photodynamic inactivation of recombinant bioluminescent Escherichia coli by cationic porphyrins under artificial and solar irradiation  

Microsoft Academic Search

A faster and simpler method to monitor the photoinactivation process of Escherichia coli involving the use of recombinant bioluminescent bacteria is described here. Escherichia coli cells were transformed with luxCDABE genes from the marine bioluminescent bacterium Vibrio fischeri and the recombinant bioluminescent indicator strain was used to assess, in real time, the effect of three cationic meso-substituted porphyrin derivatives on

Eliana Alves; Carla M. B. Carvalho; João P. C. Tomé; Maria A. F. Faustino; Maria G. P. M. S. Neves; Augusto C. Tomé; José A. S. Cavaleiro; Ângela Cunha; Sónia Mendo; Adelaide Almeida

2008-01-01

350

The human Pif1 helicase, a potential Escherichia coli RecD homologue, inhibits telomerase activity  

E-print Network

act as an adjuvant therapy for the treatment of human cancer (17). In most human tumor cells, telomereThe human Pif1 helicase, a potential Escherichia coli RecD homologue, inhibits telomerase activity is a potential homologue of Escherichia coli RecD, an ATP-dependent 50 to 30 DNA helicase. Ectopic expression

Tian, Weidong

351

Escherichia coli Bacteriocins: Antimicrobial Efficacy and Prevalence among Isolates from Patients with Bacteraemia  

Microsoft Academic Search

Bacteriocins are antimicrobial peptides generally active against bacteria closely related to the producer. Escherichia coli produces two types of bacteriocins, colicins and microcins. The in vitro efficacy of isolated colicins E1, E6, E7, K and M, was assessed against Escherichia coli strains from patients with bacteraemia of urinary tract origin. Colicin E7 was most effective, as only 13% of the

Maruška Budi?; Matija Rijavec; Živa Petkovšek; Darja Žgur-Bertok

2011-01-01

352

Structure of the Heme d of Penicillium vitale and Escherichia coli Catalases*  

E-print Network

Structure of the Heme d of Penicillium vitale and Escherichia coli Catalases* (Received-hydroxychlorin -spirolactone has been found in the crystal structures of Penicillium vitale catalase and Escherichia coli catalase hydroperoxidase II (HPII). The absolute stereochemistry of the two heme d chiral car- bon atoms

353

The RecA proteins of Deinococcus radiodurans and Escherichia coli promote DNA strand exchange  

E-print Network

-binding properties of the Ec RecA protein, and led us to examine the pathway for Dr RecA-mediated DNA strand exchangeThe RecA proteins of Deinococcus radiodurans and Escherichia coli promote DNA strand exchange via of Escherichia coli, and all filament-forming homologues identified to date, promote DNA strand exchange

Cox, Michael M.

354

MOLECULAR PROCESSING OF REPLICATION INTERMEDIATES IN ESCHERICHIA COLI AFTER DNA DAMAGE  

E-print Network

, such as dnaB266. We measured the amount of nascent DNA degradation, allowing us to determine how the newlyMOLECULAR PROCESSING OF REPLICATION INTERMEDIATES IN ESCHERICHIA COLI AFTER DNA DAMAGE By Jerilyn OF REPLICATION INTERMEDIATES IN ESCHERICHIA COLI AFTER DNA DAMAGE By Jerilyn Jalana Belle Approved

Courcelle, Justin

355

Review article Escherichia coli as a pathogen in dogs and cats  

E-print Network

Review article Escherichia coli as a pathogen in dogs and cats Lothar Beutin Robert Koch; accepted 17December 1998) Abstraet-Certain strains of Escherichia coli behave as pathogens in dogs and cats were clearly associated with enteric disease in young dogs. ETEC isolates from diar- rhoeic dogs were

Paris-Sud XI, Université de

356

Inhibition of the ATPase activity of Escherichia coli ATP synthase by magnesium fluoride  

E-print Network

Inhibition of the ATPase activity of Escherichia coli ATP synthase by magnesium fluoride Zulfiqar activity of Escherichia coli ATP synthase by magnesium fluoride (MgFx) was studied. Wild-type F1-ATPase synthesis mechanism; Magnesium fluoride; ATPase inhibition; Transition state analog 1. Introduction ATP

Zulfiqar Ahmad

357

Escherichia coli from urine of female patients with urinary tract infections is competent for intracellular bacterial community formation.  

PubMed

Nearly 50% of women experience at least one urinary tract infection (UTI) in their lifetime. Studies with mice have revealed that uropathogenic Escherichia coli (UPEC) isolates invade superficial umbrella cells that line the bladder, allowing them to find a safe haven and subvert clearance by innate host responses. Rapid intracellular replication results in the formation of distinctive intracellular bacterial communities (IBCs). In this study, we evaluated whether UPEC strains cultured from the urine of women and classified as causing acute cystitis, recurrent cystitis, asymptomatic bacteriuria, or pyelonephritis could progress through the IBC cascade in a well-characterized mouse model of cystitis. Of 18 UPEC isolates collected from women, 15 formed IBCs. Variations in the size, number, and kinetics of IBC formation were observed with strains isolated from women with different clinical syndromes. Two of the three isolates that did not form IBCs when inoculated alone were able to do so when coinoculated with an isolate that was capable of generating IBCs. The mixed infections dramatically altered the behavior of the coinfecting bacteria relative to their behavior in a single infection. The study also showed that mice with five different genetic backgrounds can support IBC formation. Although UPEC isolates differ genetically in their virulence factors, the majority of UPEC isolates from different types of UTI proceed through the IBC pathway, confirming the generality of IBCs in UTI pathogenesis in mice. PMID:17074856

Garofalo, Corinne K; Hooton, Thomas M; Martin, Steven M; Stamm, Walter E; Palermo, Joseph J; Gordon, Jeffrey I; Hultgren, Scott J

2007-01-01

358

Escherichia coli from Urine of Female Patients with Urinary Tract Infections Is Competent for Intracellular Bacterial Community Formation?  

PubMed Central

Nearly 50% of women experience at least one urinary tract infection (UTI) in their lifetime. Studies with mice have revealed that uropathogenic Escherichia coli (UPEC) isolates invade superficial umbrella cells that line the bladder, allowing them to find a safe haven and subvert clearance by innate host responses. Rapid intracellular replication results in the formation of distinctive intracellular bacterial communities (IBCs). In this study, we evaluated whether UPEC strains cultured from the urine of women and classified as causing acute cystitis, recurrent cystitis, asymptomatic bacteriuria, or pyelonephritis could progress through the IBC cascade in a well-characterized mouse model of cystitis. Of 18 UPEC isolates collected from women, 15 formed IBCs. Variations in the size, number, and kinetics of IBC formation were observed with strains isolated from women with different clinical syndromes. Two of the three isolates that did not form IBCs when inoculated alone were able to do so when coinoculated with an isolate that was capable of generating IBCs. The mixed infections dramatically altered the behavior of the coinfecting bacteria relative to their behavior in a single infection. The study also showed that mice with five different genetic backgrounds can support IBC formation. Although UPEC isolates differ genetically in their virulence factors, the majority of UPEC isolates from different types of UTI proceed through the IBC pathway, confirming the generality of IBCs in UTI pathogenesis in mice. PMID:17074856

Garofalo, Corinne K.; Hooton, Thomas M.; Martin, Steven M.; Stamm, Walter E.; Palermo, Joseph J.; Gordon, Jeffrey I.; Hultgren, Scott J.

2007-01-01

359

Pathogenic Escherichia coli in rural household container waters.  

PubMed

Plastic containers in the range of 5-20 L are widely used - especially in rural African settings - to collect, transport and store water for domestic use, including drinking, bathing and hygiene. The pathogen content of the waters in these containers has not been adequately characterized as yet. This paper presents the primary findings of a synoptic survey of drinking water quality samples from these containers and involved collection of bacterial indicator and pathogenicity gene data. In total, 571 samples of a variety of waters were taken in rural communities in South Africa and the Escherichia coli numbers measured. Of the E. coli positive samples, 46% (n = 148) were screened for the presence of E. coli pathogen gene markers. Though synoptic, the survey provided many insights into the issues that drove the study. Container use markedly degraded water quality as judged by indicator counts, even where improved water supply services were in place. Household container use also appeared to promote regrowth or contamination of containers with pathogenic E. coli strains. Polymerase chain reaction (PCR) analysis also showed that the diversity of potential pathogenic E. coli carrying virulence genes was great. All seven genes screened for (Ial, Stx1, Stx2, EaeA, Eagg, ST, LT) were found in the waters, alone or as mixtures (number of different combinations = 31) including those characteristic of the more dangerous invasive and haemorrhagic E. coli strains. Given the central role of containers in the management of water supply to rural communities, it is clear the microbiology of these waters requires much further characterization. PMID:23508146

Jagals, P; Barnard, T G; Mokoena, M M; Ashbolt, N; Roser, D J

2013-01-01

360

The Modular Organization of Protein Interactions in Escherichia coli  

PubMed Central

Escherichia coli serves as an excellent model for the study of fundamental cellular processes such as metabolism, signalling and gene expression. Understanding the function and organization of proteins within these processes is an important step towards a ‘systems’ view of E. coli. Integrating experimental and computational interaction data, we present a reliable network of 3,989 functional interactions between 1,941 E. coli proteins (?45% of its proteome). These were combined with a recently generated set of 3,888 high-quality physical interactions between 918 proteins and clustered to reveal 316 discrete modules. In addition to known protein complexes (e.g., RNA and DNA polymerases), we identified modules that represent biochemical pathways (e.g., nitrate regulation and cell wall biosynthesis) as well as batteries of functionally and evolutionarily related processes. To aid the interpretation of modular relationships, several case examples are presented, including both well characterized and novel biochemical systems. Together these data provide a global view of the modular organization of the E. coli proteome and yield unique insights into structural and evolutionary relationships in bacterial networks. PMID:19798435

Peregrín-Alvarez, José M.; Xiong, Xuejian; Su, Chong; Parkinson, John

2009-01-01

361

A second DNA methyltransferase repair enzyme in Escherichia coli.  

PubMed Central

The Escherichia coli ada-alkB operon encodes a 39-kDa protein (Ada) that is a DNA-repair methyltransferase and a 27-kDa protein (AlkB) of unknown function. By DNA blot hybridization analysis we show that the alkylation-sensitive E. coli mutant BS23 [Sedgwick, B. & Lindahl, T. (1982) J. Mol. Biol. 154, 169-175] is a deletion mutant lacking the entire ada-alkB operon. Despite the absence of the ada gene and its product, the cells contain detectable levels of a DNA-repair methyltransferase activity. We conclude that the methyltransferase in BS23 cells is the product of a gene other than ada. A similar activity was detected in extracts of an ada-10::Tn10 insertion mutant of E. coli AB1157. This DNA methyltransferase has a molecular mass of about 19 kDa and transfers the methyl groups from O6-methylguanine and O4-methylthymine in DNA, but not those from methyl phosphotriester lesions. This enzyme was not induced by low doses of alkylating agent and is expressed at low levels in ada+ and a number of ada- E. coli strains. Images PMID:3283737

Rebeck, G W; Coons, S; Carroll, P; Samson, L

1988-01-01

362

Enteroaggregative Escherichia coli associated with a foodborne outbreak of gastroenteritis.  

PubMed

This study investigated two foodborne outbreaks of gastroenteritis that occurred 10 days apart among individuals who had meals at the restaurant of a farm holiday resort. Mild gastrointestinal symptoms were reported and none of the patients needed hospitalization. Mean incubation times were 45 and 33 h, and the overall attack rates were 43.5 and 58.3%, respectively. Stool sample examination was negative for common enteric pathogens in both outbreaks. Specimens from 13 people involved in the second outbreak and 3 restaurant staff were examined for diarrhoeagenic Escherichia coli. An enteroaggregative E. coli (EAEC) strain of serotype O92:H33 was isolated from six participants and one member of staff. In particular, the EAEC strain was isolated from five of the six cases of diarrhoea examined. The strain showed an aggregative pattern of adherence to HEp-2 cells, did not produce a biofilm and possessed the virulence-related genes aat, aggR, aap and set1A, but not the astA gene. A retrospective cohort study indicated a pecorino cheese made with unpasteurized sheep milk as the possible source (P<0.001). Samples of the cheese had E. coli counts higher than 10(6) c.f.u. g(-1), but the outbreak EAEC strain was not isolated. This report confirms that EAEC infections are probably underdiagnosed because of the limited availability of laboratories capable of identifying this group of pathogenic E. coli. PMID:18719185

Scavia, Gaia; Staffolani, Monica; Fisichella, Stefano; Striano, Gianluca; Colletta, Stefano; Ferri, Giovanni; Escher, Martina; Minelli, Fabio; Caprioli, Alfredo

2008-09-01

363

Low intensity infrared laser induces filamentation in Escherichia coli cells  

NASA Astrophysics Data System (ADS)

Low intensity continuous wave and pulsed emission modes laser is used in treating many diseases and the resulting biostimulative effect on tissues has been described, yet the photobiological basis is not well understood. The aim of this wok was to evaluate, using bacterial filamentation assay, effects of laser on Escherichia coli cultures in exponential and stationary growth phase. E. coli cultures, proficient and deficient on DNA repair, in exponential and stationary growth phase, were exposed to low intensity infrared laser, aliquots were spread onto microscopic slides, stained by Gram method, visualized by optical microscopy, photographed and percentage of bacterial filamentation were determined. Low intensity infrared laser with therapeutic fluencies and different emission modes can induce bacterial filamentation in cultures of E. coli wild type, fpg/ mutM, endonuclease III and exonuclease III mutants in exponential and stationary growth phase. This study showed induction of bacterial, filamentation in E. coli cultures expose to low intensity infrared laser and attention to laser therapy protocols, which should take into account fluencies, wavelengths, tissue conditions, and genetic characteristics of cells before beginning treatment.

Fonseca, A. S.; Presta, G. A.; Geller, M.; Paoli, F.

2011-10-01

364

Activity map of the Escherichia coli RNA polymerase bridge helix.  

PubMed

Transcription, the synthesis of RNA from a DNA template, is performed by multisubunit RNA polymerases (RNAPs) in all cellular organisms. The bridge helix (BH) is a distinct feature of all multisubunit RNAPs and makes direct interactions with several active site-associated mobile features implicated in the nucleotide addition cycle and RNA and DNA binding. Because the BH has been captured in both kinked and straight conformations in different crystals structures of RNAP, recently supported by molecular dynamics studies, it has been proposed that cycling between these conformations is an integral part of the nucleotide addition cycle. To further evaluate the role of the BH, we conducted systematic alanine scanning mutagenesis of the Escherichia coli RNAP BH to determine its contributions to activities required for transcription. Combining our data with an atomic model of E. coli RNAP, we suggest that alterations in the interactions between the BH and (i) the trigger loop, (ii) fork loop 2, and (iii) switch 2 can help explain the observed changes in RNAP functionality associated with some of the BH variants. Additionally, we show that extensive defects in E. coli RNAP functionality depend upon a single previously not studied lysine residue (Lys-781) that is strictly conserved in all bacteria. It appears that direct interactions made by the BH with other conserved features of RNAP are lost in some of the E. coli alanine substitution variants, which we infer results in conformational changes in RNAP that modify RNAP functionality. PMID:21357417

Jovanovic, Milija; Burrows, Patricia C; Bose, Daniel; Cámara, Beatriz; Wiesler, Simone; Zhang, Xiaodong; Wigneshweraraj, Sivaramesh; Weinzierl, Robert O J; Buck, Martin

2011-04-22

365

CRISPR-Cas Functional Module Exchange in Escherichia coli  

PubMed Central

ABSTRACT Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (cas) genes constitute the CRISPR-Cas systems found in the Bacteria and Archaea domains. At least in some strains they provide an efficient barrier against transmissible genetic elements such as plasmids and viruses. Two CRISPR-Cas systems have been identified in Escherichia coli, pertaining to subtypes I-E (cas-E genes) and I-F (cas-F genes), respectively. In order to unveil the evolutionary dynamics of such systems, we analyzed the sequence variations in the CRISPR-Cas loci of a collection of 131 E. coli strains. Our results show that the strain grouping inferred from these CRISPR data slightly differs from the phylogeny of the species, suggesting the occurrence of recombinational events between CRISPR arrays. Moreover, we determined that the primary cas-E genes of E. coli were altogether replaced with a substantially different variant in a minor group of strains that include K-12. Insertion elements play an important role in this variability. This result underlines the interchange capacity of CRISPR-Cas constituents and hints that at least some functional aspects documented for the K-12 system may not apply to the vast majority of E. coli strains. PMID:24473126

Almendros, Cristóbal; Mojica, Francisco J. M.; Díez-Villaseñor, César; Guzmán, Noemí M.; García-Martínez, Jesús

2014-01-01

366

Marine macroalgae as a source for osmoprotection for Escherichia coli.  

PubMed

At elevated osmolarity of the mineral medium M63, marine macroalgae constitute important osmoprotectants and nutrients sources for Escherichia coli. Growth of bacterial population (16 strains) was improved by supplementing M63 salts medium with either aqueous or ethanolic algal extracts obtained from Ascophyllum nodosum, Fucus serratus, Enteromorpha ramulosa, Ulva lactuca, and Palmaria palmata. In their presence, growth was still observed even at 1.02 M NaCl. Furthermore, the E. coli ZB400 growth in presence of whole macroalgae thalli in M63/0.85 M NaCI reached its maximum within 24 h (5 × 10(7) - 5 × 10(8) colony-forming units [CFU] per milliliter). In the presence of A. nodosum, bacterial growth was inhibited. In the same experimental conditions, ethanolic extracts improved E. coli growth significantly, because the yield reached 10(11) CFU per milliliter. Ulva lactuca and P. palmata allowed the better growth. The Dragendorff-positive compounds extracted from bacterial cells growing on each ethanolic extract exhibited an osmoprotective effect as proved by a disk-diffusion assay. On the other hand, the -onium compounds (quaternary ammonium [betaines] and tertiary sulphonium) and total free amino acid contents of U. lactuca ethanolic extracts were higher than in others. Fucaceae extracts demonstrated especially high protein content. Algal extracts constitute not only an appreciable osmoprotection source for E. coli but also nutrient sources. PMID:24185483

Ghoul, M; Minet, J; Bernard, T; Dupray, E; Cormier, M

1995-09-01

367

Metabolic Flux Responses to Pyruvate Kinase Knockout in Escherichia coli  

PubMed Central

The intracellular carbon flux distribution in wild-type and pyruvate kinase-deficient Escherichia coli was estimated using biosynthetically directed fractional 13C labeling experiments with [U-13C6]glucose in glucose- or ammonia-limited chemostats, two-dimensional nuclear magnetic resonance (NMR) spectroscopy of cellular amino acids, and a comprehensive isotopomer model. The general response to disruption of both pyruvate kinase isoenzymes in E. coli was a local flux rerouting via the combined reactions of phosphoenolpyruvate (PEP) carboxylase and malic enzyme. Responses in the pentose phosphate pathway and the tricarboxylic acid cycle were strongly dependent on the environmental conditions. In addition, high futile cycling activity via the gluconeogenic PEP carboxykinase was identified at a low dilution rate in glucose-limited chemostat culture of pyruvate kinase-deficient E. coli, with a turnover that is comparable to the specific glucose uptake rate. Furthermore, flux analysis in mutant cultures indicates that glucose uptake in E. coli is not catalyzed exclusively by the phosphotransferase system in glucose-limited cultures at a low dilution rate. Reliability of the flux estimates thus obtained was verified by statistical error analysis and by comparison to intracellular carbon flux ratios that were independently calculated from the same NMR data by metabolic flux ratio analysis. PMID:11741855

Emmerling, Marcel; Dauner, Michael; Ponti, Aaron; Fiaux, Jocelyne; Hochuli, Michel; Szyperski, Thomas; Wüthrich, Kurt; Bailey, J. E.; Sauer, Uwe

2002-01-01

368

Characterization of a second lysine decarboxylase isolated from Escherichia coli.  

PubMed Central

We report here on the existence of a new gene for lysine decarboxylase in Escherichia coli K-12. The hybridization experiments with a cadA probe at low stringency showed that the homologous region of cadA was located in lambda Kohara phage clone 6F5 at 4.7 min on the E. coli chromosome. We cloned the 5.0-kb HindIII fragment of this phage clone and sequenced the homologous region of cadA. This region contained a 2,139-nucleotide open reading frame encoding a 713-amino-acid protein with a calculated molecular weight of 80,589. Overexpression of the protein and determination of its N-terminal amino acid sequence defined the translational start site of this gene. The deduced amino acid sequence showed 69.4% identity to that of lysine decarboxylase encoded by cadA at 93.7 min on the E. coli chromosome. In addition, the level of lysine decarboxylase activity increased in strains carrying multiple copies of the gene. Therefore, the gene encoding this lysine decarboxylase was designated Idc. Analysis of the lysine decarboxylase activity of strains containing cadA, ldc, or cadA ldc mutations indicated that ldc was weakly expressed under various conditions but is a functional gene in E. coli. PMID:9226257

Kikuchi, Y; Kojima, H; Tanaka, T; Takatsuka, Y; Kamio, Y

1997-01-01

369

Three-dimensional solution structure of Escherichia coli periplasmic cyclophilin.  

PubMed

The solution structure of the periplasmic cyclophilin type cis-trans peptidyl-prolyl isomerase from Escherichia coli (167 residues, MW > 18.200) has been determined using multidimensional heteronuclear NMR spectroscopy and distance geometry calculations. The structure determination is based on a total of 1720 NMR-derived restraints (1566 distance and 101 phi and 53 chi 1 torsion angle restraints). Twelve distance geometry structures were calculated, and the average root-mean-square (rms) deviation about the mean backbone coordinate positions is 0.84 +/- 0.18 A for the backbone atoms of residues 5-165 of the ensemble. The three-dimensional structure of E. coli cyclophilin consists of an eight-stranded antiparallel beta-sheet barrel capped by alpha-helices. The average coordinates of the backbone atoms of the core residues of E. coli cyclophilin have an rms deviation of 1.44 A, with conserved regions in the crystal structure of unligated human T cell cyclophilin [Ke, H. (1992) J. Mol. Biol. 228, 539-550]. Four regions proximal to the active site differ substantially and may determine protein substrate specificity, sensitivity to cyclosporin A, and the composite drug:protein surface required to inhibit calcineurin. A residue essential for isomerase activity in human T cell cyclophilin (His126) is replaced by Tyr122 in E. coli cyclophilin without affecting enzymatic activity. PMID:8130188

Clubb, R T; Ferguson, S B; Walsh, C T; Wagner, G

1994-03-15

370

Structural analysis of full-length Hfq from Escherichia coli  

PubMed Central

The structure of full-length host factor Q? (Hfq) from Escherichia coli obtained from a crystal belonging to space group P1, with unit-cell parameters a = 61.91, b = 62.15, c = 81.26?Å, ? = 78.6, ? = 86.2, ? = 59.9°, was solved by molecular replacement to a resolution of 2.85?Å and refined to R work and R free values of 20.7% and 25.0%, respectively. Hfq from E. coli has previously been crystallized and the structure has been solved for the N-terminal 72 amino acids, which cover ?65% of the full-length sequence. Here, the purification, crystallization and structural data of the full 102-amino-acid protein are presented. These data revealed that the presence of the C-terminus changes the crystal packing of E. coli Hfq. The crystal structure is discussed in the context of the recently published solution structure of Hfq from E. coli. PMID:21543856

Beich-Frandsen, Mads; Ve?erek, Branislav; Sjöblom, Björn; Bläsi, Udo; Djinovi?-Carugo, Kristina

2011-01-01

371

Antimicrobial resistance patterns of uropathogens among children in Istanbul, Turkey.  

PubMed

Urinary tract infections are a common cause of end-stage renal disease in Turkey. This prospective study investigated the antibiotic resistance patterns of uropathogens in order to recommend appropriate therapeutic protocols for children with urinary tract infections in Istanbul, Turkey. Between October 2007 and October 2008, children presenting with a first episode of urinary tract infection to a pediatric outpatient clinic were enrolled in the study. Urine samples were cultured, and antimicrobial susceptibility testing was performed. Children with proven urinary tract infections underwent imaging studies where available. A total of 126 children with a first episode of community-acquired urinary tract infection were enrolled in the study. The median age was 60.6 months; 84.1% of the children were female. Of the 126 urine samples, Escherichia coli was the leading uropathogen (81.7%), followed by Proteus spp (7.1%), Klebsiella spp (4.0%), Enterococcus spp (3.2%), Enterobacter spp (2.4%), and Pseudomonas spp (1.6%). Among the isolated uropathogens, resistance to ampicillin (85.0%), amoxicillin-clavulanate (73.8%), cefazolin (37.3%) and trimethoprim-sulfamethoxazole (42.9%) was remarkable. A large number of Enterococcus species were resistant to all antimicrobial agents except vancomycin. A country-based evaluation of antibiotic susceptibility is needed to modify antibiotic treatment. Resistance to antimicrobial agents commonly used to treat urinary tract infections (nitrofurantoin, cefixime) is less a problem than resistance to other antimicrobials (aminopenicillins, cephalosporins, trimethoprim-sulfamethoxazole) frequently prescribed for other indications. PMID:21710858

Ipek, Ilke Ozahi; Bozaykut, Abdulkadir; Arman, Didem Caktir; Sezer, Rabia Gonul

2011-03-01

372

A second prepilin peptidase gene in Escherichia coli K-12.  

PubMed

Escherichia coli K-12 strains grown at 37 degrees C or 42 degrees C, but not at 30 degrees C, process the precursors of the Neisseria gonorrhoeae type IV pilin PilE and the Klebsiella oxytoca type IV pseudopilin PulG in a manner reminiscent of the prepilin peptidase-dependent processing of these proteins that occurs in these bacteria. Processing of prePulG in Escherichia coli requires a glycine at position -1, as does processing by the cognate prepilin peptidase (PulO), and is unaffected by mutations that inactivate several non-specific proteases. These data suggested that E. coli K-12 has a functional prepilin peptidase, despite the fact that it does not itself appear to express either type IV pilin or pseudopilin genes under the conditions that allow prePilE and prePulG processing. The E. coli K-12 genome contains two genes encoding proteins with significant sequence similarity to prepilin peptidases: gspO at minute 74.5 and pppA (f310c) at minute 67 on the genetic map. We have previously obtained evidence that gspO encodes an active enzyme but is not transcribed. pppA was cloned and shown to code for a functional prepilin peptidase capable of processing typical prepilin peptidase substrates. Inactivation of pppA eliminated the endogenous, thermoinducible prepilin peptidase activity. PppA was able to replace PulO prepilin peptidase in a pullulanase secretion system reconstituted in E. coli when expressed from high-copy-number plasmids but not when present in a single chromosomal copy. The analysis of pppA-lacZ fusions indicated that pppA expression was very low and regulated by the growth temperature at the level of translation, in agreement with the observed temperature dependence of PppA activity. Polymerase chain reaction and Southern hybridization analyses revealed the presence of the pppA gene in 12 out of 15 E. coli isolates. PMID:9515702

Franceti?, O; Lory, S; Pugsley, A P

1998-02-01

373

Characterization of recombinant human chymase expressed in Escherichia coli.  

PubMed

We compared recombinant human chymase expressed in Escherichia coli with human chymase purified from vascular tissues. The recombinant chymase, the structure of which was NH2-enterokinase cleavage site-chymase-COOH, was expressed in Escherichia coli and then was solubilized and renatured. The protein did not have a chymase activity, but gained this activity after the cleavage of the N-terminal site by enterokinase. The enzyme was purified by heparin affinity and gel filtration columns. The N-terminal sequence of the protein was identical to the sequence for human chymase. The molecular weights of the recombinant chymase and chymase purified from human vascular tissues were 26 and 30 kDa, respectively, and the 4 kDa difference was thought to be due to the presence or absence of glycan. The optimum pH of the recombinant enzyme activity was between 7.5 and 9.0. The activity of the recombinant enzyme was inhibited by chymostatin, soybean trypsin inhibitor and phenylmethylsulfonyl fluoride, but not by ethylenediaminetetraacetic acid and aprotinin. This enzyme cleaved specifically the Phe8-His9 bond of angiotensin (Ang) I to form Ang II and that of big endothelin (ET)-1 to form ET-1-(1-31). These findings demonstrated that the enzymatic characteristics of the recombinant enzyme were identical to that of native human chymase. PMID:10877533

Takai, S; Sumi, S; Aoike, M; Sakaguchi, M; Itoh, Y; Jin, D; Matsumura, E; Miyazaki, M

2000-02-01

374

Fluoroquinolone levels in healthy dog urine following a 20-mg/kg oral dose of enrofloxacin exceed mutant prevention concentration targets against Escherichia coli isolated from canine urinary tract infections.  

PubMed

A 3-day course of oral enrofloxacin is effective for treating uncomplicated urinary tract infection (UTI) in dogs when administered 20 mg/kg Q24H. However, emergence of fluoroquinolone-resistant mutants of uropathogens is a concern. Urine concentrations of enrofloxacin and ciprofloxacin were measured in six healthy dogs following dose of enrofloxacin 20 mg/kg. Mutant prevention concentrations of Escherichia coli isolated from canine UTI were also determined against ciprofloxacin. Urine AUC(24)/MPC ratios considering ciprofloxacin concentrations ranged 3819-7767, indicating that selection of resistant E. coli mutants in dogs with uncomplicated UTIs is unlikely in the bladder given that an AUC(24)/MPC = 39 is considered to be protective against mutant selection for ciprofloxacin. However, additional studies are required to evaluate the effects of this enrofloxacin treatment protocol on bacteria that colonize anatomic sites where fluoroquinolones achieve lower concentrations compared to the urinary bladder. PMID:23859001

Daniels, J B; Tracy, G; Irom, S J; Lakritz, J

2014-04-01

375

Characterization of Escherichia coli serotype O157:H7.  

PubMed

A total of 174 strains of Escherichia coli serotype O157:H7 representing human isolates obtained from outbreaks and sporadic cases of hemorrhagic colitis, hemolytic-uremic syndrome, and nonbloody diarrheal illnesses as well as from asymptomatic carriers across Canada and the United States were examined. E. coli serotype O157:H7 possessed distinct biochemical markers, a 100% negative reaction for beta-glucuronidase and sorbitol, and a 100% positive reaction for raffinose and dulcitol; all strains otherwise were biochemically typical of E. coli. The vast majority (97%) of the strains were susceptible to commonly used antimicrobial agents. All strains produced readily detectable levels of Verotoxin; however, with polymyxin extraction, nearly 50% of the strains showed up to a 10-fold increase in the toxin level. None were found to mediate hemagglutination of human group A erythrocytes with or without D-mannose. The majority (approximately 70%) of the strains showed localized and diffuse adherence to HEp-2 cells and Henle 407 cells, and the adherence patterns were not very different from those observed among other E. coli strains. Twenty phage types were recognized, with phage types 1 and 2 accounting for 65% of the test strains. Plasmid analysis indicated three basic plasmid profiles: profile I was characterized by 68.7- and 4.2-megadalton (MDa) plasmids (62% of strains), profile II was characterized by 66.2- and 1.8-MDa plasmids (20% of strains), and profile III was characterized by a 62.5-MDa plasmid (18% of strains). A small number (19%) of the strains carried at least one additional plasmid over the basic complements, and these could be considered to constitute a miscellaneous category. None of the above-described characteristics of E. coli serotype O157:H7 could be directly correlated with one another, with the nature of infection, or with the geographical distribution of strains. PMID:3053758

Ratnam, S; March, S B; Ahmed, R; Bezanson, G S; Kasatiya, S

1988-10-01

376

Reduction of Escherichia coli adherence to uroepithelial bladder cells after consumption of cranberry juice: a double-blind randomized placebo-controlled cross-over trial.  

PubMed

To determine the efficacy of the consumption of cranberry juice versus placebo with regard to the presence of in vitro bacterial anti-adherence activity in the urine of healthy volunteers. Twenty healthy volunteers, 10 men and 10 women, were included. The study was a double-blind, randomized, placebo-controlled, and cross-over study. In addition to normal diet, each volunteer received at dinner a single dose of 750 ml of a total drink composed of: (1) 250 ml of the placebo and 500 ml of mineral water, or (2) 750 ml of the placebo, or (3) 250 ml of the cranberry juice and 500 ml of mineral water, or (4) 750 ml of the cranberry juice. Each volunteer took the four regimens successively in a randomly order, with a washout period of at least 6 days between every change in regimen. The first urine of the morning following cranberry or placebo consumption was collected and used to support bacterial growth. Six uropathogenic Escherichia coli strains (all expressing type 1 pili; three positive for the gene marker for P-fimbriae papC and three negative for papC), previously isolated from patients with symptomatic urinary tract infections, were grown in urine samples and tested for their ability to adhere to the T24 bladder cell line in vitro. There were no significant differences in the pH or specific gravity between the urine samples collected after cranberry or placebo consumption. We observed a dose dependent significant decrease in bacterial adherence associated with cranberry consumption. Adherence inhibition was observed independently from the presence of genes encoding type P pili and antibiotic resistance phenotypes. Cranberry juice consumption provides significant anti-adherence activity against different E. coli uropathogenic strains in the urine compared with placebo. PMID:16397814

Di Martino, P; Agniel, R; David, K; Templer, C; Gaillard, J L; Denys, P; Botto, H

2006-02-01

377

Phenotypic and genotypic properties of Escherichia coli isolated from colisepticemic cases of Japanese quail  

Microsoft Academic Search

This study was conducted to characterize the Escherichia coli isolates from colisepticemic Japanese quails. One hundred and nine E. coli were isolated in pure culture from heart blood of dead Japanese quails. The sampled birds were originated from four different\\u000a farms. Antibiotic resistance pattern of E. coli isolates were determined against nine antibacterial agents. Phylotype and virulence genes of the

Mahmood Salehi; Reza Ghanbarpour

2010-01-01

378

Mouse in vivo neutralization of Escherichia coli Shiga toxin 2 with monoclonal antibodies  

Technology Transfer Automated Retrieval System (TEKTRAN)

Escherichia coli (E. coli) food contaminations pose serious health and food safety concerns, and have been the subject of massive food recalls. Shiga toxin 2 (Stx2)-producing E. coli has been identified as the major cause of hemorrhagic colitis and hemolytic uremic syndrome (HUS), the most severe di...

379

Localization of protein aggregation in Escherichia coli is governed by diffusion and nucleoid macromolecular crowding effect  

E-print Network

1 Localization of protein aggregation in Escherichia coli is governed by diffusion and nucleoid. coli is purely diffusive (Brownian). Using single-particle tracking of protein aggregates in live E. coli cells, we estimated the average size and diffusion constant of the aggregates. Our results

Paris-Sud XI, Université de

380

A homolog of an Escherichia coli phosphate-binding protein gene from Xanthomonas oryzae pv. oryzae  

NASA Technical Reports Server (NTRS)

A Xanthomonas oryzae pv. oryzae gene with sequence similarity to an Escherichia coli phosphate-binding protein gene (phoS) produces a periplasmic protein of apparent M(r) 35,000 when expressed in E. coli. Amino terminal sequencing revealed that a signal peptide is removed during transport to the periplasm in E. coli.

Hopkins, C. M.; White, F. F.; Heaton, L. A.; Guikema, J. A.; Leach, J. E.; Spooner, B. S. (Principal Investigator)

1995-01-01

381

Finished Genome Sequence of Escherichia coli K-12 Strain HMS174 (ATCC 47011)  

PubMed Central

Escherichia coli strain K-12 substrain HMS174 is an engineered descendant of the E. coli K-12 wild-type strain. Like its ancestor, it is an important organism in biotechnological research and is used in fermentation processes for heterologous protein production. Here, we report the complete genome sequence of E. coli HMS174 (ATCC 47011). PMID:25414489

Mairhofer, Juergen; Krempl, Peter M.; Thallinger, Gerhard G.

2014-01-01

382

ON-CHIP PARALLEL DETECTION OF SHIGA TOXIN PRODUCING ESCHERICHIA COLI USING  

E-print Network

ON-CHIP PARALLEL DETECTION OF SHIGA TOXIN PRODUCING ESCHERICHIA COLI USING LOOP-MEDIATED ISOTHERMAL AMPLIFICATION Carlos Duarte Guevara, Rashid Bashir Shiga toxin-producing Escheria coli (STEC) strains amplification based detection of E.coli using a system that creates a nano-droplet array and then amplify target

Bashir, Rashid

383

SENSITIVE DETECTION OF ESCHERICHIA COLI 0157:H7 BY THE USE OF IMMUNOMAGNETIC AND FLUORESCENT BEADS  

Technology Transfer Automated Retrieval System (TEKTRAN)

To meet the needs of food safety, a rapid and sensitive fluorescent sandwich method for the detection of Escherichia coli O157:H7 in ground beef was developed. Immunomagnetic beads (IMBs) coated with anti-E. coli O157:H7 antibodies were used to capture and concentrate E. coli O157:H7 present in gro...

384

Diet, Escherichia coli 0157:H7, and cattle, a review after 10 years  

Technology Transfer Automated Retrieval System (TEKTRAN)

Escherichia coli are commensal bacteria that can account for up to 1% of the bacterial population of the gut. Ruminant animals are reservoirs of the pathogenic bacteria E. coli strain O157:H7 and approximately 30% of feedlot cattle shed E. coli O157:H7. Feedlot and high-producing dairy cattle are ...

385

Recombinant expression of bioactive peptide lunasin in Escherichia coli.  

PubMed

Lunasin, a cancer-preventive peptide, was isolated from soybean, barley, and wheat. Previous studies showed that this 43-amino acid peptide has the ability to suppress chemical carcinogen-induced transformation in mammalian cells and skin carcinogenesis in mice. In this study, we attempted to use the Escherichia coli T7 expression system for expression of lunasin. The lunasin gene was synthesized by overlapping extension polymerase chain reaction and expressed in E. coli BL21(DE3) with the use of vector pET29a. The recombinant lunasin containing his-tag at the C-terminus was expressed in soluble form which could be purified by immobilized metal affinity chromatography. After 4 h, the expression level is above 4.73 mg of recombinant his-tagged lunasin/L of Luria-Bertani broth. It does not affect the bacterial growth and expression levels. This is the first study that successfully uses E. coli as a host to produce valuable bioactive lunasin. The result of in vitro bioassay showed that the purified recombinant lunasin can inhibit histone acetylation. Recombinant lunasin also inhibits the release of pro-inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta, and nitric oxide production). Compared with other research methods on extraction or chemical synthesis to produce lunasin, our method is very efficient in saving time and cost. In the future, it could be applied in medicine and structure-function determination. PMID:20625716

Liu, Chin-Feng; Pan, Tzu-Ming

2010-09-01

386

The binary protein-protein interaction landscape of Escherichia coli.  

PubMed

Efforts to map the Escherichia coli interactome have identified several hundred macromolecular complexes, but direct binary protein-protein interactions (PPIs) have not been surveyed on a large scale. Here we performed yeast two-hybrid screens of 3,305 baits against 3,606 preys (?70% of the E. coli proteome) in duplicate to generate a map of 2,234 interactions, which approximately doubles the number of known binary PPIs in E. coli. Integration of binary PPI and genetic-interaction data revealed functional dependencies among components involved in cellular processes, including envelope integrity, flagellum assembly and protein quality control. Many of the binary interactions that we could map in multiprotein complexes were informative regarding internal topology of complexes and indicated that interactions in complexes are substantially more conserved than those interactions connecting different complexes. This resource will be useful for inferring bacterial gene function and provides a draft reference of the basic physical wiring network of this evolutionarily important model microbe. PMID:24561554

Rajagopala, Seesandra V; Sikorski, Patricia; Kumar, Ashwani; Mosca, Roberto; Vlasblom, James; Arnold, Roland; Franca-Koh, Jonathan; Pakala, Suman B; Phanse, Sadhna; Ceol, Arnaud; Häuser, Roman; Siszler, Gabriella; Wuchty, Stefan; Emili, Andrew; Babu, Mohan; Aloy, Patrick; Pieper, Rembert; Uetz, Peter

2014-03-01

387

Engineered biosynthesis of medium-chain esters in Escherichia coli.  

PubMed

Medium-chain esters such as isobutyl acetate (IBAc) and isoamyl acetate (IAAc) are high-volume solvents, flavors and fragrances. In this work, we engineered synthetic metabolic pathways in Escherichia coli for the total biosynthesis of IBAc and IAAc directly from glucose. Our pathways harnessed the power of natural amino acid biosynthesis. In particular, the native valine and leucine pathways in E. coli were utilized to supply the precursors. Then alcohol acyltransferases from various organisms were investigated on their capability to catalyze esterification reactions. It was discovered that ATF1 from Saccharomyces cerevisiae was the best enzyme for the formation of both IBAc and IAAc in E. coli. In vitro biochemical characterization of ATF1 confirmed the fermentation results and provided rational guidance for future enzyme engineering. We also performed strain improvement by removing byproduct pathways (?ldh, ?poxB, ?pta) and increased the production of both target chemicals. Then the best IBAc producing strain was used for scale-up fermentation in a 1.3-L benchtop bioreactor. 36g/L of IBAc was produced after 72h fermentation. This work demonstrates the feasibility of total biosynthesis of medium-chain esters as renewable chemicals. PMID:25447641

Tai, Yi-Shu; Xiong, Mingyong; Zhang, Kechun

2015-01-01

388

Characterization of Pyruvate Uptake in Escherichia coli K-12  

PubMed Central

The monocarboxylate pyruvate is an important metabolite and can serve as sole carbon source for Escherichia coli. Although specific pyruvate transporters have been identified in two bacterial species, pyruvate transport is not well understood in E. coli. In the present study, pyruvate transport was investigated under different growth conditions. The transport of pyruvate shows specific activities depending on the growth substrate used as sole carbon source, suggesting the existence of at least two systems for pyruvate uptake: i) one inducible system and probably highly specific for pyruvate and ii) one system active under non-induced conditions. Using the toxic pyruvate analog 3-fluoropyruvate, a mutant was isolated unable to grow on and transport pyruvate. Further investigation revealed that a revertant selected for growth on pyruvate regained the inducible pyruvate transport activity. Characterization of pyruvate excretion showed that the pyruvate transport negative mutant accumulated pyruvate in the growth medium suggesting an additional transport system for pyruvate excretion. The here presented data give valuable insight into the pyruvate metabolism and transport of E. coli suggesting the presence of at least two uptake systems and one excretion system to balance the intracellular level of pyruvate. PMID:23818977

Kreth, Jens; Lengeler, Joseph W.; Jahreis, Knut

2013-01-01

389

Fluorine-19 nuclear magnetic resonance studies of Escherichia coli membranes.  

PubMed Central

Several fluorinated fatty acids of the general structure CH3(CH2)13--mCF2(CH2)m--2COOH are incorporated biosynthetically as unsaturated fatty acid analogues into the phospholipids of Escherichia coli. Under optimum conditions an unsaturated fatty acid autotroph, K1060B5, can be grown so that 50% of the total phospholipid fatty acids are 8,8-difluoromyristate. Conditions are found for which more than 20% of the fatty acids are fluorinated before a decrease in growth rate is observed. We have used 19F nuclear magnetic resonance to examine membranes isolated from E. coli grown under the latter conditions. A comparison is made with spectra of aqueous dispersions of extracted E. coli phospholipids and model multilayer phospholipid membranes. An explanation of the 19F resonance line shape in these membrane systems and the relationship to a molecular order parameter is given. It is apparent that 19F nuclear magnetic resonance is more sensitive to the degree of ordering or fluidity of phospholipids than spin labels or fluorescent probes. For instance, a dramatic effect of membrane protein on lipid fluidity can be seen. Finally, this method can be used to measure the proportion of frozen and fluid lipid in biological membranes at temperatures within the span of the gel-to-lipid phase transition. PMID:345274

Gent, M P; Cottam, P F; Ho, C

1978-01-01

390

Fitness, Stress Resistance, and Extraintestinal Virulence in Escherichia coli  

PubMed Central

The extraintestinal virulence of Escherichia coli is dependent on numerous virulence genes. However, there is growing evidence for a role of the metabolic properties and stress responses of strains in pathogenesis. We assessed the respective roles of these factors in strain virulence by developing phenotypic assays for measuring in vitro individual and competitive fitness and the general stress response, which we applied to 82 commensal and extraintestinal pathogenic E. coli strains previously tested in a mouse model of sepsis. Individual fitness properties, in terms of maximum growth rates in various media (Luria-Bertani broth with and without iron chelator, minimal medium supplemented with gluconate, and human urine) and competitive fitness properties, estimated as the mean relative growth rate per generation in mixed cultures with a reference fluorescent E. coli strain, were highly diverse between strains. The activity of the main general stress response regulator, RpoS, as determined by iodine staining of the colonies, H2O2 resistance, and rpoS sequencing, was also highly variable. No correlation between strain fitness and stress resistance and virulence in the mouse model was found, except that the maximum growth rate in urine was higher for virulent strains. Multivariate analysis showed that the number of virulence factors was the only independent factor explaining the virulence in mice. At the species level, growth capacity and stress resistance are heterogeneous properties that do not contribute significantly to the intrinsic virulence of the strains. PMID:23690401

Bleibtreu, Alexandre; Gros, Pierre-Alexis; Laouénan, Cédric; Clermont, Olivier; Le Nagard, Hervé; Picard, Bertrand; Tenaillon, Olivier

2013-01-01

391

Binding of type 1-piliated Escherichia coli to vaginal mucus.  

PubMed Central

To better understand the interactions involved in bacterial adherence and the role of mucus in the pathogenesis of urinary tract infections, we developed a system to study the binding of a recombinant Escherichia coli strain, HB101/pWRS1-17, expressing type 1 pili, to vaginal mucus collected from 28 women. Bacteria bound to differing extents to all specimens examined, and preincubation of bacteria with mannose inhibited binding by 50 to 89%. Additionally, all mucus samples showed reactivity with anti-mannose antibody, and the levels of reactivity correlated with the levels of bacterial binding, suggesting that the mannose-terminal saccharides present on these glycoproteins are the receptors for the binding of type 1-piliated bacteria. Mucus specimens collected over periods of 5 days and 12 weeks exhibited significant variation in bacterial binding, indicating temporal differences in the ability of vaginal mucus to act as a receptor for type 1-piliated E. coli. The results show that vaginal mucus can bind bacteria and may thus influence the initial attachment and subsequent colonization of the vaginal and urinary tract epithelium by E. coli. PMID:7822005

Venegas, M F; Navas, E L; Gaffney, R A; Duncan, J L; Anderson, B E; Schaeffer, A J

1995-01-01

392

Colibri: a functional data base for the Escherichia coli genome.  

PubMed Central

Several data libraries have been created to organize all the data obtained worldwide about the Escherichia coli genome. Because the known data now amount to more than 40% of the whole genome sequence, it has become necessary to organize the data in such a way that appropriate procedures can associate knowledge produced by experiments about each gene to its position on the chromosome and its relation to other relevant genes, for example. In addition, global properties of genes, affected by the introduction of new entries, should be present as appropriate description fields. A data base, implemented on Macintosh by using the data base management system 4th Dimension, is described. It is constructed around a core constituted by known contigs of E. coli sequences and links data collected in general libraries (unmodified) to data associated with evolving knowledge (with modifiable fields). Biologically significant results obtained through the coupling of appropriate procedures (learning or statistical data analysis) are presented. The data base is available through a 4th Dimension runtime and through FTP on Internet. It has been regularly updated and will be systematically linked to other E. coli data bases (M. Kroger, R. Wahl, G. Schachtel, and P. Rice, Nucleic Acids Res. 20(Suppl.):2119-2144, 1992; K. E. Rudd, W. Miller, C. Werner, J. Ostell, C. Tolstoshev, and S. G. Satterfield, Nucleic Acids Res. 19:637-647, 1991) in the near future. Images PMID:8246843

Médigue, C; Viari, A; Hénaut, A; Danchin, A

1993-01-01

393

Characterization of the YdeO Regulon in Escherichia coli  

PubMed Central

Enterobacteria are able to survive under stressful conditions within animals, such as acidic conditions in the stomach, bile salts during transfer to the intestine and anaerobic conditions within the intestine. The glutamate-dependent (GAD) system plays a major role in acid resistance in Escherichia coli, and expression of the GAD system is controlled by the regulatory cascade consisting of EvgAS > YdeO > GadE. To understand the YdeO regulon in vivo, we used ChIP-chip to interrogate the E. coli genome for candidate YdeO binding sites. All of the seven operons identified by ChIP-chip as being potentially regulated by YdeO were confirmed as being under the direct control of YdeO using RT-qPCR, EMSA, DNaseI-footprinting and reporter assays. Within this YdeO regulon, we identified four stress-response transcription factors, DctR, NhaR, GadE, and GadW and enzymes for anaerobic respiration. Both GadE and GadW are involved in regulation of the GAD system and NhaR is an activator for the sodium/proton antiporter gene. In conjunction with co-transcribed Slp, DctR is involved in protection against metabolic endoproducts under acidic conditions. Taken all together, we suggest that YdeO is a key regulator of E. coli survival in both acidic and anaerobic conditions. PMID:25375160

Yamanaka, Yuki; Oshima, Taku; Ishihama, Akira; Yamamoto, Kaneyoshi

2014-01-01

394

Potential production platform of n-butanol in Escherichia coli.  

PubMed

We proposed a potential production platform of n-butanol in Escherichia coli. First, a butyrate-conversion strain was developed by removal of undesired genes and recruiting endogenous atoDA and Clostridium adhE2. Consequently, this E. coli strain grown on the M9 mineral salt with yeast extract (M9Y) was shown to produce 6.2g/L n-butanol from supplemented butyrate at 36h. The molar conversion yield of n-butanol on butyrate reaches 92%. Moreover, the production platform was advanced by additional inclusion of a butyrate-producing strain. This strain was equipped with a pathway comprising atoDA and heterologous genes for the synthesis of butyrate. Without butyrate, the butyrate-conversion and the butyrate-producing strains were co-cultured in M9Y medium and produced 5.5g/L n-butanol from glucose at 24h. The production yield on glucose accounts for 69% of the theoretical yield. Overall, it indicates a promise of the developed platform for n-butanol production in E. coli. PMID:25461833

Saini, Mukesh; Hong Chen, Min; Chiang, Chung-Jen; Chao, Yun-Peng

2015-01-01

395

Identification of Diarrheagenic Escherichia coli Strains from Avian Organic Fertilizers  

PubMed Central

The Brazilian poultry industry generates large amounts of organic waste, such as chicken litter, which is often used in agriculture. Among the bacteria present in organic fertilizer are members of the Enterobacteriaceae family. The objective of this study was to detect the presence of diarrheagenic Escherichia coli (DEC) strains in avian organic fertilizer, and assess the potential damage they can cause in humans due to antimicrobial resistance. The presence of DEC pathotypes and phylogenetic groups were detected by multiplex-PCR. Phenotypic assays, such as tests for adhesion, cytotoxicity activity, biofilm formation and especially antimicrobial susceptibility, were performed. Fifteen DEC strains from 64 E. coli were isolated. Among these, four strains were classified as enteropathogenic (EPEC; 6.2%), three strains as Shiga toxin-producing (STEC; 4.7%), 10 strains as enteroaggregative (EAEC; 12.5%), but two of these harbored the eaeA gene too. The low number of isolated strains was most likely due to the composting process, which reduces the number of microorganisms. These strains were able to adhere to HEp-2 and HeLa cells and produce Shiga-toxins and biofilms; in addition, some of the strains showed antimicrobial resistance, which indicates a risk of the transfer of resistance genes to human E. coli. These results showed that DEC strains isolated from avian organic fertilizers can cause human infections. PMID:25170683

Puño-Sarmiento, Juan; Gazal, Luis Eduardo; Medeiros, Leonardo P.; Nishio, Erick K.; Kobayashi, Renata K. T.; Nakazato, Gerson

2014-01-01

396

Comprehensive Mapping of the Escherichia coli Flagellar Regulatory Network  

PubMed Central

Flagellar synthesis is a highly regulated process in all motile bacteria. In Escherichia coli and related species, the transcription factor FlhDC is the master regulator of a multi-tiered transcription network. FlhDC activates transcription of a number of genes, including some flagellar genes and the gene encoding the alternative Sigma factor FliA. Genes whose expression is required late in flagellar assembly are primarily transcribed by FliA, imparting temporal regulation of transcription and coupling expression to flagellar assembly. In this study, we use ChIP-seq and RNA-seq to comprehensively map the E. coli FlhDC and FliA regulons. We define a surprisingly restricted FlhDC regulon, including two novel regulated targets and two binding sites not associated with detectable regulation of surrounding genes. In contrast, we greatly expand the known FliA regulon. Surprisingly, 30 of the 52 FliA binding sites are located inside genes. Two of these intragenic promoters are associated with detectable noncoding RNAs, while the others either produce highly unstable RNAs or are inactive under these conditions. Together, our data redefine the E. coli flagellar regulatory network, and provide new insight into the temporal orchestration of gene expression that coordinates the flagellar assembly process. PMID:25275371

Fitzgerald, Devon M.; Bonocora, Richard P.; Wade, Joseph T.

2014-01-01

397

Virulence genes of Escherichia coli strains isolated from mastitic milk.  

PubMed

Escherichia coli, a Gram-negative environmental pathogen associated with bovine mastitis was isolated from the milk of 34 symptomatic cows that had been diagnosed with clinical mastitis. Eighty isolates were obtained over a 17-month period and these isolates were screened by DNA amplification for the following E. coli virulence genes: cnf1, cnf2, eaeA, eagg, einv, ltx1, stx1, stx2 and vt2e. Thirty of the bacterial isolates, obtained from 23 different cows, had toxin genes identified in their DNA. The most common virulence gene detected was stx1, with a prevalence of 31%, followed by cnf2 (7.5%), vt2e (6.25%) and eaeA (4%). The possession of different virulence genes by the bacterial isolates had no discernable impact on the health status of the cows as there was no correlation between the potential for toxin production by the E. coli isolates and the systemic clinical condition of the respective infected cows. PMID:15458491

Bean, A; Williamson, J; Cursons, R T

2004-08-01

398

Epithelial cell invasion by bovine septicemic Escherichia coli.  

PubMed Central

Little is known regarding the pathogenesis of Escherichia coli-induced septicemic colibacillosis of calves. To understand the mechanism by which these strains penetrate the intestinal epithelium and gain access to the bloodstream, we examined the potential of bovine septicemic E. coli to invade cultured epithelial cells. By using a gentamicin survival assay, we demonstrated bacterial invasion of Madin-Darby canine kidney (MDCK) cells. Transcytosis of polarized MDCK cell monolayers was also observed, but only when bacteria were added to the basolateral surface. Electron microscopy confirmed the presence of intracellular organisms which appeared to be within membrane-bound vacuoles. The bovine septicemic isolate used in this study expressed the fimbrial adhesion CS31A. To examine the role of CS31A-mediated adherence in invasion and transcytosis of MDCK cell monolayers, a CS31A-deficient mutant was constructed by suicide vector-mediated insertional mutagenesis. Although nonadherent, the mutant showed a level of invasion similar to that of the wild-type parent. E. coli DH5 alpha carrying the cloned CS31A determinant was noninvasive. These findings suggest that expression of CS31A is neither required nor sufficient to mediate invasion. Images PMID:7903284

Korth, M J; Lara, J C; Moseley, S L

1994-01-01

399

Rapid microarray-based DNA genoserotyping of Escherichia coli.  

PubMed

In this study, an improvement in the oligonucleotide-based DNA microarray for the genoserotyping of Escherichia coli is presented. Primer and probes for additional 70 O antigen groups were developed. The microarray was transferred to a new platform, the ArrayStrip format, which allows high through-put tests in 96-well formats and fully automated microarray analysis. Thus, starting from a single colony, it is possible to determine within a few hours and a single experiment, 94 of the over 180 known O antigen groups as well as 47 of the 53 different H antigens. The microarray was initially validated with a set of defined reference strains that had previously been serotyped by conventional agglutination in various reference centers. For further validation of the microarray, 180 clinical E. coli isolates of human origin (from urine samples, blood cultures, bronchial secretions, and wound swabs) and 53 E. coli isolates from cattle, pigs, and poultry were used. A high degree of concordance between the results of classical antibody-based serotyping and DNA-based genoserotyping was demonstrated during validation of the new 70 O antigen groups as well as for the field strains of human and animal origin. Therefore, this oligonucleotide array is a diagnostic tool that is user-friendly and more efficient than classical serotyping by agglutination. Furthermore, the tests can be performed in almost every routine lab and are easily expanded and standardized. PMID:24298918

Geue, Lutz; Monecke, Stefan; Engelmann, Ines; Braun, Sascha; Slickers, Peter; Ehricht, Ralf

2014-02-01

400

Regulation of neutrophil phagocytosis of Escherichia coli by antipsychotic drugs.  

PubMed

Antipsychotic drugs (APDs) have been used to ease the symptoms of schizophrenia. APDs have recently been reported to regulate the immune response. Our previous studies revealed that the atypical APDs risperidone and clozapine and the typical APD haloperidol can inhibit the phagocytic ability of macrophages. Our research next determined the effects of APDs on the phagocytic ability of neutrophils, which are the most abundant type of white blood cells in mammals. Here we provide evidence that clozapine and haloperidol can induce increased phagocytic uptake of Escherichia coli by differentiated HL-60 cells and by purified human neutrophils. Furthermore, clozapine and haloperidol can increase the myeloperoxidase activity and IL-8 production in neutrophils. Our results also show that clozapine can inhibit E. coli survival within differentiated HL-60 cells. Furthermore, clozapine and haloperidol are shown to enhance cell surface Mac-1 expression and the activated AKT signaling pathway in purified neutrophils exposed to E. coli. These results indicate that clozapine and haloperidol can increase the phagocytic ability of neutrophils by increasing AKT activation when cells are exposed to bacteria. PMID:25448498

Chen, Mao-Liang; Wu, Semon; Tsai, Tzung-Chieh; Wang, Lu-Kai; Tsai, Fu-Ming

2014-12-01

401

[Escherichia coli virulence factors causing peritonitis, appendicitis and other extraintestinal infections].  

PubMed

Fifty-nine E. coli strains isolated from clinical cases of peritonitis, appendicitis, cholecystitis, wounds and respiratory infections as well as from other miscellaneous sources were investigated. A control group constituted by 475 E. coli strains isolated from the faeces of healthy individuals were also studied. E. coli O-grouped and investigated for production of cytotoxic necrotizing factor CNF1 and alpha-haemolysin (Hly), expression of P fimbriae and mannose-resistant (MRHA) and mannose-sensitive (MSHA) haemagglutination. Virulence factors significantly associated with extraintestinal strains were: production of CNF1 (19% versus 5%, p < 0.001), Hly (27% versus 9%; p < 0.001) and expression of MRHA (44% versus 16%; p < 0.001). The majority of extraintestinal strains (68% versus 36%; p < 0.001), in contrast with faecal E. coli, belonged to O serogroups frequently detected in uropathogenic and bacteraemic E. coli. These results suggest that E. coli causing different types of extraintestinal infections show similar virulence factors and belong to the same serogroups. However, between E. coli isolated from intraabdominal, wound and respiratory infections the number of strains with virulence factors was lower than in E. coli causing urinary tract infections and sepsis. PMID:1450257

Blanco, J; Blanco, J E; Alonso, M P; Blanco, M; Garabal, J I; González, E A

1992-01-01

402

Escherichia coli Strains Isolated from the Uteri Horn, Mouth, and Rectum of Bitches Suffering from Pyometra: Virulence Factors, Antimicrobial Susceptibilities, and Clonal Relationships among Strains.  

PubMed

Pyometra is recognized as one of the main causes of disease and death in the bitch, and Escherichia coli is the major pathogen associated with this disease. In this study, 70 E. coli isolates from the uteri horn, mouth, and rectum of bitches suffering from the disease and 43 E. coli isolates from the rectum of clinically healthy bitches were examined for the presence of uropathogenic virulence genes and susceptibility to antimicrobial drugs. DNA profiles of isolates from uteri horn and mouth in bitches with pyometra were compared by REP, ERIC, and BOX-PCR. Virulence gene frequencies detected in isolates from canine pyometra were as follows: 95.7% fim, 27.1% iss, 25.7% hly, 18.5% iuc, and 17.1% usp. Predominant resistance was determined for cephalothin, ampicillin, and nalidixic acid among the isolates from all sites examined. Multidrug resistance was found on ? 50% pyometra isolates. Using the genotypic methods some isolates from uteri, pus, and saliva of the same bitch proved to have identical DNA profiles which is a reason for concern due to the close relationship between household pets and humans. PMID:24734047

Agostinho, Juliana M A; de Souza, Andressa; Schocken-Iturrino, Ruben P; Beraldo, Lívia G; Borges, Clarissa A; Avila, Fernando A; Marin, José M

2014-01-01

403

Escherichia coli Strains Isolated from the Uteri Horn, Mouth, and Rectum of Bitches Suffering from Pyometra: Virulence Factors, Antimicrobial Susceptibilities, and Clonal Relationships among Strains  

PubMed Central

Pyometra is recognized as one of the main causes of disease and death in the bitch, and Escherichia coli is the major pathogen associated with this disease. In this study, 70 E. coli isolates from the uteri horn, mouth, and rectum of bitches suffering from the disease and 43 E. coli isolates from the rectum of clinically healthy bitches were examined for the presence of uropathogenic virulence genes and susceptibility to antimicrobial drugs. DNA profiles of isolates from uteri horn and mouth in bitches with pyometra were compared by REP, ERIC, and BOX-PCR. Virulence gene frequencies detected in isolates from canine pyometra were as follows: 95.7% fim, 27.1% iss, 25.7% hly, 18.5% iuc, and 17.1% usp. Predominant resistance was determined for cephalothin, ampicillin, and nalidixic acid among the isolates from all sites examined. Multidrug resistance was found on ?50% pyometra isolates. Using the genotypic methods some isolates from uteri, pus, and saliva of the same bitch proved to have identical DNA profiles which is a reason for concern due to the close relationship between household pets and humans. PMID:24734047