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Sample records for coli k1 strains

  1. The Escherichia coli argW-dsdCXA genetic island is highly variable, and E. coli K1 strains commonly possess two copies of dsdCXA.

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    Moritz, Rebecca L; Welch, Rodney A

    2006-11-01

    The genome sequences of Escherichia coli pathotypes reveal extensive genetic variability in the argW-dsdCXA island. Interestingly, the archetype E. coli K1 neonatal meningitis strain, strain RS218, has two copies of the dsdCXA genes for d-serine utilization at the argW and leuX islands. Because the human brain contains d-serine, an epidemiological study emphasizing K1 isolates surveyed the dsdCXA copy number and function. Forty of 41 (97.5%) independent E. coli K1 isolates could utilize d-serine. Southern blot hybridization revealed physical variability within the argW-dsdC region, even among 22 E. coli O18:K1:H7 isolates. In addition, 30 of 41 K1 strains, including 21 of 22 O18:K1:H7 isolates, had two dsdCXA loci. Mutational analysis indicated that each of the dsdA genes is functional in a rifampin-resistant mutant of RS218, mutant E44. The high percentage of K1 strains that can use d-serine is in striking contrast to our previous observation that only 4 of 74 (5%) isolates in the diarrheagenic E. coli (DEC) collection have this activity. The genome sequence of diarrheagenic E. coli isolates indicates that the csrRAKB genes for sucrose utilization are often substituted for dsdC and a portion of dsdX present at the argW-dsdCXA island of extraintestinal isolates. Among DEC isolates there is a reciprocal pattern of sucrose fermentation versus d-serine utilization. The ability to use d-serine is a trait strongly selected for among E. coli K1 strains, which have the ability to infect a wide range of extraintestinal sites. Conversely, diarrheagenic E. coli pathotypes appear to have substituted sucrose for d-serine as a potential nutrient.

  2. Comparative genomic analysis shows that avian pathogenic Escherichia coli isolate IMT5155 (O2:K1:H5; ST complex 95, ST140 shares close relationship with ST95 APEC O1:K1 and human ExPEC O18:K1 strains.

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    Xiangkai Zhu Ge

    Full Text Available Avian pathogenic E. coli and human extraintestinal pathogenic E. coli serotypes O1, O2 and O18 strains isolated from different hosts are generally located in phylogroup B2 and ST complex 95, and they share similar genetic characteristics and pathogenicity, with no or minimal host specificity. They are popular objects for the study of ExPEC genetic characteristics and pathogenesis in recent years. Here, we investigated the evolution and genetic blueprint of APEC pathotype by performing phylogenetic and comparative genome analysis of avian pathogenic E. coli strain IMT5155 (O2:K1:H5; ST complex 95, ST140 with other E. coli pathotypes. Phylogeny analyses indicated that IMT5155 has closest evolutionary relationship with APEC O1, IHE3034, and UTI89. Comparative genomic analysis showed that IMT5155 and APEC O1 shared significant genetic overlap/similarities with human ExPEC dominant O18:K1 strains (IHE3034 and UTI89. Furthermore, the unique PAI I5155 (GI-12 was identified and found to be conserved in APEC O2 serotype isolates. GI-7 and GI-16 encoding two typical T6SSs in IMT5155 might be useful markers for the identification of ExPEC dominant serotypes (O1, O2, and O18 strains. IMT5155 contained a ColV plasmid p1ColV5155, which defined the APEC pathotype. The distribution analysis of 10 sequenced ExPEC pan-genome virulence factors among 47 sequenced E. coli strains provided meaningful information for B2 APEC/ExPEC-specific virulence factors, including several adhesins, invasins, toxins, iron acquisition systems, and so on. The pathogenicity tests of IMT5155 and other APEC O1:K1 and O2:K1 serotypes strains (isolated in China through four animal models showed that they were highly virulent for avian colisepticemia and able to cause septicemia and meningitis in neonatal rats, suggesting zoonotic potential of these APEC O1:K1 and O2:K1 isolates.

  3. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii trophozoites and cysts.

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    Matin, Abdul; Jung, Suk-Yul

    2011-12-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. coli K12 was also applied. The association ratio of E. coli K1 with A. castellanii was 4.3 cfu per amoeba for 1 hr but E. coli K5 with A. castellanii was 1 cfu per amoeba for 1 hr. By invasion and survival assays, E. coli K5 was recovered less than E. coli K1 but still alive inside A. castellanii. E. coli K1 and K5 survived and multiplied intracellularly in A. castellanii. The survival assay was performed under a favourable condition for 22 hr and 43 hr with the encystment of A. castellanii. Under the favourable condition for the transformation of trophozoites into cysts, E. coli K5 multiplied significantly. Moreover, the pathogenic potential of E. coli K1 from A. castellanii cysts exhibited no changes as compared with E. coli K1 from A. castellanii trophozoites. E. coli K5 was multiplied in A. castellanii trophozoites and survived in A. castellanii cysts. Therefore, this study suggests that E. coli K5 can use A. castellanii as a reservoir host or a vector for the bacterial transmission.

  4. Interaction of Escherichia coli K1 and K5 with Acanthamoeba castellanii Trophozoites and Cysts

    OpenAIRE

    Matin, Abdul; Jung, Suk-Yul

    2011-01-01

    The existence of symbiotic relationships between Acanthamoeba and a variety of bacteria is well-documented. However, the ability of Acanthamoeba interacting with host bacterial pathogens has gained particular attention. Here, to understand the interactions of Escherichia coli K1 and E. coli K5 strains with Acanthamoeba castellanii trophozoites and cysts, association assay, invasion assay, survival assay, and the measurement of bacterial numbers from cysts were performed, and nonpathogenic E. ...

  5. The Plasmid of Escherichia coli Strain S88 (O45:K1:H7) That Causes Neonatal Meningitis Is Closely Related to Avian Pathogenic E. coli Plasmids and Is Associated with High-Level Bacteremia in a Neonatal Rat Meningitis Model▿

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    Peigne, Chantal; Bidet, Philippe; Mahjoub-Messai, Farah; Plainvert, Céline; Barbe, Valérie; Médigue, Claudine; Frapy, Eric; Nassif, Xavier; Denamur, Erick; Bingen, Edouard; Bonacorsi, Stéphane

    2009-01-01

    A new Escherichia coli virulent clonal group, O45:K1, belonging to the highly virulent subgroup B21 was recently identified in France, where it accounts for one-third of E. coli neonatal meningitis cases. Here we describe the sequence, epidemiology and function of the large plasmid harbored by strain S88, which is representative of the O45:K1 clonal group. Plasmid pS88 is 133,853 bp long and contains 144 protein-coding genes. It harbors three different iron uptake systems (aerobactin, salmochelin, and the sitABCD genes) and other putative virulence genes (iss, etsABC, ompTP, and hlyF). The pS88 sequence is composed of several gene blocks homologous to avian pathogenic E. coli plasmids pAPEC-O2-ColV and pAPEC-O1-ColBM. PCR amplification of 11 open reading frames scattered throughout the plasmid was used to investigate the distribution of pS88 and showed that a pS88-like plasmid is present in other meningitis clonal groups such as O18:K1, O1:K1, and O83:K1. A pS88-like plasmid was also found in avian pathogenic strains and human urosepsis strains belonging to subgroup B21. A variant of S88 cured of its plasmid displayed a marked loss of virulence relative to the wild-type strain in a neonatal rat model, with bacteremia more than 2 log CFU/ml lower. The salmochelin siderophore, a known meningovirulence factor, could not alone explain the plasmid's contribution to virulence, as a salmochelin mutant displayed only a minor fall in bacteremia (0.9 log CFU/ml). Thus, pS88 is a major virulence determinant related to avian pathogenic plasmids that has spread not only through meningitis clonal groups but also human urosepsis and avian pathogenic strains. PMID:19307211

  6. Probiotic Mixture Golden Bifido Prevents Neonatal Escherichia coli K1 Translocation via Enhancing Intestinal Defense

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    Qing Zeng

    2017-09-01

    Full Text Available Escherichia coli (E. coli K1 sepsis and meningitis is a severe infection characterized by high mortality in neonates. Successful colonization and translocation across the intestinal mucosa have been regarded as the critical steps for E. coli K1 sepsis and meningitis. We recently reported that the probiotic mixture, Golden Bifido (containing live Lactobacillus bulgaricus, Bifidobacterium, and Streptococcus thermophilus, LBS has a preventive role against neonatal E. coli K1 bacteremia and meningitis. However, the interaction between the neonatal gut barrier, probiotics and E. coli K1 is still not elucidated. The present study aims to investigate how LBS exerts its protective effects on neonatal gut barrier during E. coli K1 infection. The beneficial effects of LBS were explored in vitro and in vivo using human colon carcinoma cell lines HT-29 and rat model of neonatal E. coli K1 infection, respectively. Our results showed that stimulation with E. coli K1 was able to cause intestinal barrier dysfunction, which were reflected by E. coli K1-induced intestinal damage and apoptosis of intestinal epithelial cells, reduction of mucin, immunoglobulin A (IgA and tight junction proteins expression, as well as increase in intestinal permeability, all these changes facilitate E. coli K1 intestinal translocation. However, these changes were alleviated when HT-29 cells were treated with LBS before E. coli K1 infection. Furthermore, we found that LBS-treated neonatal rats (without E. coli K1 infection have showed higher production of mucin, ZO-1, IgA, Ki67 in intestinal mucosa as well as lower intestinal permeability than that of non-treated rats, indicating that LBS could accelerate the development of neonatal intestinal defense. Taken together, our results suggest that enhancement of the neonatal intestinal defense to fight against E. coli K1 translocation could be the potential mechanism to elucidate how LBS confers a protective effect against neonatal E

  7. Whole-Genome Sequences of the Archetypal K1 Escherichia coli Neonatal Isolate RS218 and Contemporary Neonatal Bacteremia Clinical Isolates SCB11, SCB12, and SCB15.

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    Day, Michael W; Jackson, Lydgia A; Akins, Darrin R; Dyer, David W; Chavez-Bueno, Susana

    2015-02-26

    Neonatal bacteremia Escherichia coli strains commonly belong to the K1 capsular type. Their ability to cause invasive neonatal disease appears to be determined by other virulence factors that have yet to be identified. We report here the genome sequences of four E. coli neonatal bacteremia isolates, including that of the archetypal strain RS218.

  8. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

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    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-12-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent.

  9. Conservation of plasmids among Escherichia coli K1 isolates of diverse origins.

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    Mercer, A A; Morelli, G; Heuzenroeder, M; Kamke, M; Achtman, M

    1984-01-01

    Escherichia coli K1 isolates of various O types were previously assigned to different clonal groups. Members of the two clones defined by membrane pattern 9 (MP9) and serotypes O18:K1 and O1:K1 had been found to be very similar to each other. The plasmid contents of these bacteria confirmed this conclusion. Both groups carried a self-transmissible plasmid of the FI incompatibility group that coded for colicin production and a major outer membrane protein called the plasmid-coded protein (PCP). The size of this plasmid varied from 76 to 96 megadaltons, but restriction endonuclease digestion and DNA heteroduplex analysis revealed that these plasmids were highly related. O18:K1 bacteria of MP6 had previously been determined to represent a subclone, related to but different from O18:K1 MP9 bacteria. These MP6 bacteria carried a different, smaller IncFI plasmid which did not code for colicin production or the PCP protein. This smaller plasmid was primarily related to the larger plasmid within the regions of DNA encoding incompatibility, replication, and conjugation. O1:K1 bacteria of MP5 contained other unrelated plasmids in agreement with the previous conclusion that they are unrelated to O1:K1 bacteria of MP9. The bacteria examined had been isolated from two continents over a time span of 38 years, and the results attest to conservative inheritance of plasmids within bacteria of common descent. Images PMID:6094355

  10. Asymptomatic bacteriuria Escherichia coli strains

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    Hancock, Viktoria; Nielsen, E.M.; Klemm, Per

    2006-01-01

    Urinary tract infections (UTIs) affect millions of people each year. Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU) in humans. Persons affected by ABU may carry a particular E. coli strain for extended periods of time without any symptoms. In contrast...

  11. Asymptomatic bacteriuria Escherichia coli strains

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Nielsen, E.M.; Klemm, Per

    2006-01-01

    Urinary tract infections (UTIs) affect millions of people each year. Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU) in humans. Persons affected by ABU may carry a particular E. coli strain for extended periods of time without any symptoms. In contrast...

  12. Going for baroque at the Escherichia coli K1 cell surface.

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    King, Michael R; Steenbergen, Susan M; Vimr, Eric R

    2007-05-01

    Phase variation is usually thought of as the stochastic switching between alternatively expressed ('on') and unexpressed ('off') phenotypic states. However, coupling synthesis of a monotonous homopolysaccharide to a mechanism of random but incomplete chemical modification produces almost infinite structural variation. Potentially limitless variability implies that evolution can produce highly ornate or extravagant flourishes reminiscent of the baroque style. Here, we describe an analysis of capsular polysialic acid form variation in Escherichia coli K1, demonstrating that the large number of variant structures is controlled by a single contingency locus. The mechanism for generating maximum structural diversity from maximal genetic parsimony is conferred by a simple translational switch carried on a K1-specific prophage.

  13. Emergence of Carbapenem-Resistant Serotype K1 Hypervirulent Klebsiella pneumoniae Strains in China.

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    Zhang, Rong; Lin, Dachuan; Chan, Edward Wai-Chi; Gu, Danxia; Chen, Gong-Xiang; Chen, Sheng

    2015-11-16

    We report the emergence of five carbapenem-resistant K1 hypervirulent Klebsiella pneumoniae (hvKP) strains which caused fatal infections in hospital patients in Zhejiang Province, China, upon entry through surgical wounds. Genotyping results revealed the existence of three genetically related strains which exhibited a new sequence type, ST1797, and revealed that all strains harbored the magA and wcaG virulence genes and a plasmid-borne bla(KPC-2) gene. These findings indicate that K1 hvKP is simultaneously hypervirulent, multidrug resistant, and transmissible.

  14. Lactobacillus rhamnosus GG supernatant enhance neonatal resistance to systemic Escherichia coli K1 infection by accelerating development of intestinal defense

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    He, Xiaolong; Zeng, Qing; Puthiyakunnon, Santhosh; Zeng, Zhijie; Yang, Weijun; Qiu, Jiawen; Du, Lei; Boddu, Swapna; Wu, Tongwei; Cai, Danxian; Huang, Sheng-He; Cao, Hong

    2017-01-01

    The objective of this study was to determine whether Lactobacillus rhamnosus GG culture supernatant (LCS) has a preventive effect against gut-derived systemic neonatal Escherichia coli (E. coli) K1 infection. The preventive effects were evaluated in human colonic carcinoma cell line Caco-2 and neonatal rat models. Our in vitro results showed that LCS could block adhesion, invasion and translocation of E. coli K1 to Caco-2 monolayer via up-regulating mucin production and maintaining intestinal integrity. In vivo experiments revealed that pre-treatment with LCS significantly decrease susceptibility of neonatal rats to oral E. coli K1 infection as reflected by reduced bacterial intestinal colonization, translocation, dissemination and systemic infections. Further, we found that LCS treated neonatal rats have higher intestinal expressions of Ki67, MUC2, ZO-1, IgA, mucin and lower barrier permeability than those in untreated rats. These results indicated that LCS could enhance neonatal resistance to systemic E. coli K1 infection via promoting maturation of neonatal intestinal defense. In conclusions, our findings suggested that LCS has a prophylactic effect against systemic E. coli K1 infection in neonates. Future studies aimed at identifying the specific active ingredients in LCS will be helpful in developing effective pharmacological strategies for preventing neonatal E. coli K1 infection. PMID:28262688

  15. Bioluminescent imaging reveals novel patterns of colonization and invasion in systemic Escherichia coli K1 experimental infection in the neonatal rat.

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    Witcomb, Luci A; Collins, James W; McCarthy, Alex J; Frankel, Gadi; Taylor, Peter W

    2015-12-01

    Key features of Escherichia coli K1-mediated neonatal sepsis and meningitis, such as a strong age dependency and development along the gut-mesentery-blood-brain course of infection, can be replicated in the newborn rat. We examined temporal and spatial aspects of E. coli K1 infection following initiation of gastrointestinal colonization in 2-day-old (P2) rats after oral administration of E. coli K1 strain A192PP and a virulent bioluminescent derivative, E. coli A192PP-lux2. A combination of bacterial enumeration in the major organs, two-dimensional bioluminescence imaging, and three-dimensional diffuse light imaging tomography with integrated micro-computed tomography indicated multiple sites of colonization within the alimentary canal; these included the tongue, esophagus, and stomach in addition to the small intestine and colon. After invasion of the blood compartment, the bacteria entered the central nervous system, with restricted colonization of the brain, and also invaded the major organs, in line with increases in the severity of symptoms of infection. Both keratinized and nonkeratinized surfaces of esophagi were colonized to a considerably greater extent in susceptible P2 neonates than in corresponding tissues from infection-resistant 9-day-old rat pups; the bacteria appeared to damage and penetrate the nonkeratinized esophageal epithelium of infection-susceptible P2 animals, suggesting the esophagus represents a portal of entry for E. coli K1 into the systemic circulation. Thus, multimodality imaging of experimental systemic infections in real time indicates complex dynamic patterns of colonization and dissemination that provide new insights into the E. coli K1 infection of the neonatal rat. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

  16. ppk1 gene relates with the pathogenesis of meningitis infected by E.coli K1%脑膜炎大肠杆菌K1株ppk1基因致病机制初探

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    彭亮; 赵铁; 罗文英; 付美芹; 曹虹; 黄胜和

    2012-01-01

    [Objective] To construct the polyphosphate kinase 1 gene deletion mutant of Es-cherichia coli (E. Coli) Kl strain E44 and explore the role of ppkl in the pathopoiesis of meningitis E. Coli Kl. [Methods] The ppkl gene was knocked out from the genome of E. Coli Kl strain E44 by using suicide vetor pCVD442 and homologous recombination, and the mutant was named Appkl. Then the survival ablities of E44 and Appkl in poor nutrition condition and oxidative stress were examined. The ability of the mutant adhering to human brain microvas-cular endothelial cells (HBMEC) was also compared with that of the wild type. The cytoge-netic toxic effects induced by Appkl and E44 were tested by using the lactic dehydrogenase testing kit. [Results] The ppkl gene of the mutant had been knocked out and was confirmed by PCR and DNA sequencing. The ppkl deletion mutant showed to be defective in adhesion ability to HBMEC and survival abilities under poor nutrition condition and oxidative stress. The damaging effects of HBMEC induced by Appkl were significantly less than that induced by E44. [Conclusion] The ppkl gene plays an important role in E. Coli Kl surviving in low nutrition and oxidative stress condition, adhering to HBMEC and inducing cell injury.%[目的]构建脑膜炎大肠杆菌K1 (Escherichia coli,E.coli K1)株E44的聚磷酸盐激酶1 (Polyphosphate kinase 1,PPK1)基因敲除株,并对其生物学功能进行初步研究,为明确ppk1基因在E.coli K1株致脑膜炎机制中的作用奠定基础.[方法]利用自杀质粒pCVD442及基因同源重组技术敲除E.coli K1株E44中的ppk1基因,构建ppk1缺失突变株Δppk1;体外比较野生株和突变株在低营养及氧化压力情况下的生存能力;考察二者对人脑微血管内皮细胞(Human brain microvascular endothelial cells,HBMEC)的黏附能力;通过测定乳酸脱氢酶(Lactic dehydrogenase,LDH)释放活性,比较野生株和突变株对HBMEC的损伤效应.[结果]PCR及序列分析

  17. Neonatal Escherichia coli K1 meningitis causes learning and memory impairments in adulthood.

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    Barichello, Tatiana; Dagostim, Valdemira S; Generoso, Jaqueline S; Simões, Lutiana R; Dominguini, Diogo; Silvestre, Cintia; Michels, Monique; Vilela, Márcia Carvalho; Jornada, Luciano K; Comim, Clarissa M; Dal-Pizzol, Felipe; Teixeira, Antonio Lucio; Quevedo, João

    2014-07-15

    Neonatal Escherichia coli meningitis continues to be an important cause of mortality and morbidity in newborns worldwide. The aim of this study was to investigate the cytokines/chemokines, brain-derived neurotrophic factor (BDNF) levels, blood-brain barrier integrity in neonatal rats following E. coli K1 experimental meningitis infection and subsequent behavioural parameters in adulthood. In the hippocampus, interleukin increased at 96 h, IL-6 at 12, 48 and 96 h, IL-10 at 96 h, cytokine-induced neutrophil chemoattractant-1 at 6, 12, 24, 48 and 96 h, and BDNF at 48 and 96 h. In the cerebrospinal fluid, tumour necrosis factor alpha levels increased at 6, 12, 24, 48 and 96 h. The BBB breakdown occurred at 12 h in the hippocampus, and at 6h in the cortex. We evaluated behavioural parameters in adulthood: habituation to the open-field, step-down inhibitory avoidance, object recognition, continuous multiple-trials step-down inhibitory avoidance and forced swimming tasks. In adulthood, the animals showed habituation and aversive memory impairment. The animals needed a significant increase in the number of training periods to learn and not had depressive-like symptoms.

  18. Lactobacillus rhamnosus GG Suppresses Meningitic E. coli K1 Penetration across Human Intestinal Epithelial Cells In Vitro and Protects Neonatal Rats against Experimental Hematogenous Meningitis

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    Sheng-He Huang

    2009-01-01

    Full Text Available The purpose of this study was to examine prophylactic efficacy of probiotics in neonatal sepsis and meningitis caused by E. coli K1. The potential inhibitory effect of Lactobacillus rhamnosus GG (LGG on meningitic E. coli K1 infection was examined by using (i in vitro inhibition assays with E44 (a CSF isolate from a newborn baby with E. coli meningitis, and (ii the neonatal rat model of E. coli sepsis and meningitis. The in vitro studies demonstrated that LGG blocked E44 adhesion, invasion, and transcytosis in a dose-dependent manner. A significant reduction in the levels of pathogen colonization, E. coli bacteremia, and meningitis was observed in the LGG-treated neonatal rats, as assessed by viable cultures, compared to the levels in the control group. In conclusion, probiotic LGG strongly suppresses meningitic E. coli pathogens in vitro and in vivo. The results support the use of probiotic strains such as LGG for prophylaxis of neonatal sepsis and meningitis.

  19. The clinical, pathological and microbiological outcome of an Escherichia coli O2:K1 infection in avian pneumovirus infected turkeys.

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    Van de Zande, S; Nauwynck, H; Pensaert, M

    2001-08-20

    The purpose of this study was to evaluate the effect of an Escherichia coli infection in avian pneumovirus (APV)-infected turkeys. One group of 2-week-old specific pathogen-free (SPF) and two groups of 3-week-old conventional (CON) turkeys were inoculated oculonasally with virulent APV subtype A alone, with E. coli O2:K1 alone or with both agents at varying intervals (1, 3, 5 or 7 days) between the two inoculations. The birds were followed clinically and examined for macroscopic lesions at necropsy. Titres of APV were determined in the turbinates, trachea, lungs and air sacs. The number of E. coli O2:K1were assessed in the turbinates, trachea, lungs, air sacs, liver and heart. In both SPF and CON turkeys, dual infection resulted in an increased morbidity and a higher incidence of gross lesions compared to the groups given single infections, especially with a time interval between APV and E. coli inoculations of 3 and 5 days. APV was isolated from the respiratory tract of all APV-infected groups between 3 and 7 days post inoculation. E. coli O2:K1 was isolated only from turkeys that received a dual infection. It was recovered from the turbinates, trachea, lungs, heart and liver. These results show that APV may act as a primary agent predisposing to E. coli colonization and invasion.

  20. Temporal changes of oxidative stress markers in Escherichia coli K1-induced experimental meningitis in a neonatal rat model.

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    Giridharan, Vijayasree V; Simões, Lutiana R; Dagostin, Valdemira S; Generoso, Jaqueline S; Rezin, Gislaine T; Florentino, Drielly; Muniz, Jhonata P; Collodel, Allan; Petronilho, Fabricia; Quevedo, Joao; Barichello, Tatiana

    2017-07-13

    Despite advances in antimicrobial therapy and advanced critical care neonatal bacterial meningitis has a mortality rate of over 10% and induces neurological sequelae in 20-50% of cases. Escherichia coli K1 (E. coli K1) is the most common gram-negative organism causing neonatal meningitis and is the second most common cause behind group B streptococcus. We previously reported that an E. coli K1 experimental meningitis infection in neonatal rats resulted in habituation and aversive memory impairment and a significant increase in cytokine levels in adulthood. In this present study, we investigated the oxidative stress profile including malondialdehyde (MDA) levels, carbonyl protein formation, myeloperoxidase activity (MPO) activity, superoxide dismutase (SOD) activity and catalase (CAT) activity 6, 12, 24, 48, 72 and 96h after E. coli K1 experimental meningitis infection. In addition, sulfhydryl groups, nitrite and nitrate levels and activity of the mitochondrial respiratory chain enzymes were also measured in the frontal cortex and hippocampus of neonatal rats. The results from this study demonstrated a significant increase in MDA, protein carbonyls and MPO activity and a simultaneous decrease in SOD activity in the hippocampus of the neonatal meningitis survivors but the same was not observed in frontal cortex. In addition, we also observed a significant increase in complex IV activity in the hippocampus and frontal cortex of meningitis survivor rats. Thus, the results from this study reaffirmed the possible role of oxidative stress, nitric oxide and its related compounds in the complex pathophysiology of E. coli K1-induced bacterial meningitis. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Efficacy of avian pneumovirus vaccines against an avian pneumovirus/Escherichia coli O2:K1 dual infection in turkeys.

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    Van de Zande, S; Nauwynck, H; Pensaert, M

    2002-03-16

    The clinical, pathological and microbiological outcome of a challenge with avian pneumovirus (APV) and Escherichia coli O2:K1 was evaluated in turkeys vaccinated with an attenuated APV vaccine and with or without maternally derived antibodies. Two groups of two-week-old poults, one with and one without maternally derived antibodies against APV, were vaccinated oculonasally with attenuated APV subtype A or B. A third group remained unvaccinated. Eleven weeks later, the turkeys were inoculated intranasally with either virulent APV subtype A, or E. coli O2:K1, or with both agents three days apart. After the dual infection, birds vaccinated with attenuated subtype A or B, and with or without maternally derived antibodies, had lower mean clinical scores than the unvaccinated birds. In the vaccinated birds, virus replication was significantly reduced and no bacteria were isolated, except from the birds vaccinated with attenuated subtype B. In the unvaccinated turkeys, large numbers of E. coli O2:K1 were isolated from the turbinates of the dually infected birds between one-and-a-half and seven days after they were inoculated.

  2. The asymptomatic bacteriuria Escherichia coli strain 83972 outcompetes uropathogenic E. coli strains in human urine

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Ulett, G.C.; Schembri, M.A.

    2006-01-01

    Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU). In contrast to uropathogenic E. coli (UPEC), which causes symptomatic urinary tract infections (UTI), very little is known about the mechanisms by which these strains colonize the human urinary tract....... The prototype ABU E. coli strain 83972 was originally isolated from a girl who had carried it asymptomatically for 3 years. Deliberate colonization of UTI-susceptible individuals with E. coli 83972 has been used successfully as an alternative approach for the treatment of patients who are refractory...... to conventional therapy. Colonization with strain 83972 appears to prevent infection with UPEC strains in such patients despite the fact that this strain is unable to express the primary adhesins involved in UTI, viz. P and type 1 fimbriae. Here we investigated the growth characteristics of E. coli 83972 in human...

  3. Biosynthesis of the Escherichia coli K1 group 2 polysialic acid capsule occurs within a protected cytoplasmic compartment.

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    Steenbergen, Susan M; Vimr, Eric R

    2008-06-01

    Capsular polysaccharides are important virulence determinants in a wide range of invasive infectious diseases. Although capsule synthesis has been extensively investigated, understanding polysaccharide export from the cytoplasm to the external environment has been more difficult. Here we present the results of a novel protection assay indicating that synthesis and export of the Escherichia coli K1 group 2 capsular polysialic acid (K1 antigen) occur within a protected subcellular compartment designated the sialisome. In addition to the polymerase encoded by neuS, localization and complementation analyses indicated that the sialisome includes the accessory membrane protein NeuE. The requirement for NeuE was suppressed by overproducing NeuS, suggesting that NeuE functions by stabilizing the polymerase or facilitating its assembly in the sialisome. Although an interaction between NeuE and NeuS could not be demonstrated with a bacterial two-hybrid system that reconstitutes an intracellular cell-signalling pathway, interactions between NeuS and KpsC as well as other sialisome components were detected. The combined results provide direct evidence for specific protein-protein interactions in the synthesis and export of group 2 capsular polysaccharides under in vivo conditions. The approaches developed here will facilitate further dissection of the sialisome, suggesting similar methodology for understanding the biosynthesis of other group 2 capsules.

  4. The asymptomatic bacteriuria Escherichia coli strain 83972 outcompetes uropathogenic E. coli strains in human urine

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Ulett, G.C.; Schembri, M.A.

    2006-01-01

    Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU). In contrast to uropathogenic E. coli (UPEC), which causes symptomatic urinary tract infections (UTI), very little is known about the mechanisms by which these strains colonize the human urinary tract...

  5. Cyclomodulins in urosepsis strains of Escherichia coli.

    Science.gov (United States)

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

    2010-06-01

    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, urosepsis origin (P, 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.

  6. Hydrogen production by recombinant Escherichia coli strains

    Science.gov (United States)

    Maeda, Toshinari; Sanchez‐Torres, Viviana; Wood, Thomas K.

    2012-01-01

    Summary The production of hydrogen via microbial biotechnology is an active field of research. Given its ease of manipulation, the best‐studied bacterium Escherichia coli has become a workhorse for enhanced hydrogen production through metabolic engineering, heterologous gene expression, adaptive evolution, and protein engineering. Herein, the utility of E. coli strains to produce hydrogen, via native hydrogenases or heterologous ones, is reviewed. In addition, potential strategies for increasing hydrogen production are outlined and whole‐cell systems and cell‐free systems are compared. PMID:21895995

  7. An improved high-quality draft genome sequence of Carnobacterium inhibens subsp. inhibens strain K1(T).

    Science.gov (United States)

    Nicholson, Wayne L; Davis, Christina L; Shapiro, Nicole; Huntemann, Marcel; Clum, Alicia; Reddy, T B K; Pillay, Manoj; Markowitz, Victor; Varghese, Neha; Pati, Amrita; Ivanova, Natalia; Kyrpides, Nikos; Woyke, Tanja

    2016-01-01

    Despite their ubiquity and their involvement in food spoilage, the genus Carnobacterium remains rather sparsely characterized at the genome level. Carnobacterium inhibens K1(T) is a member of the Carnobacteriaceae family within the class Bacilli. This strain is a Gram-positive, rod-shaped bacterium isolated from the intestine of an Atlantic salmon. The present study determined the genome sequence and annotation of Carnobacterium inhibens K1(T). The genome comprised 2,748,608 bp with a G + C content of 34.85 %, which included 2621 protein-coding genes and 116 RNA genes. The strain contained five contigs corresponding to presumptive plasmids of sizes: 19,036; 24,250; 26,581; 65,272; and 65,904 bp.

  8. Effects of Saponins against Clinical E. coli Strains and Eukaryotic Cell Line

    Directory of Open Access Journals (Sweden)

    Michał Arabski

    2012-01-01

    Full Text Available Saponins are detergent-like substances showing antibacterial as well as anticancer potential. In this study, the effects of saponins from Quillaja saponaria were analyzed against prokaryotic and eukaryotic cells. Multidrug-resistant clinical E. coli strains were isolated from human urine. As eukaryotic cells, the CHO-K1 cell lines were applied. Antibacterial effect of ampicillin, streptomycin, and ciprofloxacin in the presence of saponins was measured by cultivation methods. Properties of saponins against CHO-K1 cells were measured by the MTT test, hemolysis assay and flow cytometry. Saponin from Quillaja saponaria has a cytotoxic effect at concentrations higher than 25 μg/mL and in the range of 12–50 μg/mL significantly increases the level of early apoptotic cells. Saponin at dose of 12 μg/mL enhances the six E. coli strains growth. We postulate that saponins increase the influx of nutrients from the medium into E. coli cells. Saponins do not have synergetic effects on antibacterial action of tested antibiotics. In contrary, in the presence of saponins and antibiotics, more CFU/mL E. coli cells were observed. This effect was similar to saponins action alone towards E. coli cells. In conclusion, saponins was cytotoxic against CHO-K1 cells, whereas against E. coli cells this effect was not observed.

  9. Regulation of Toll-like receptor 2 interaction with Ecgp96 controls Escherichia coli K1 invasion of brain endothelial cells

    Science.gov (United States)

    Krishnan, Subramanian; Chen, Shuang; Turcatel, Gianluca; Arditi, Moshe; Prasadarao, Nemani V.

    2012-01-01

    SUMMARY The interaction of outer membrane protein A (OmpA) with its receptor, Ecgp96 (a homologue of Hsp90β) is critical for the pathogenesis of E. coli K1 meningitis. Since Hsp90 chaperones Toll-like receptors (TLRs), we examined the role of TLRs in E. coli K1 infection. Herein, we show that newborn TLR2−/− mice are resistant to E. coli K1 meningitis, while TLR4−/− mice succumb to infection sooner. In vitro, OmpA+ E. coli infection selectively upregulates Ecgp96 and TLR2 in human brain microvascular endothelial cells (HBMEC), whereas OmpA− E. coli upregulates TLR4 in these cells. Furthermore, infection with OmpA+ E. coli causes Ecgp96 and TLR2 translocate to the plasma membrane of HBMEC as a complex. Immunoprecipitation studies of the plasma membrane fractions from infected HBMEC reveal that the C-termini of Ecgp96 and TLR2 are critical for OmpA+ E. coli invasion. Knockdown of TLR2 using siRNA results in inefficient membrane translocation of Ecgp96 and significantly reduces invasion. In addition, the interaction of Ecgp96 and TLR2 induces a bipartite signal, one from Ecgp96 through PKC-α while the other from TLR2 through MyD88, ERK1/2 and NF-κB. This bipartite signal ultimately culminates in the efficient production of NO, which in turn promotes E. coli K1 invasion of HBMEC. PMID:22963587

  10. Purification and immunochemical properties of Escherichia coli B polysaccharide cross-reacting with Salmonella typhi Vi antigen: preliminary evidence for cross-reaction of the polysaccharide with Escherichia coli K1 antigen.

    OpenAIRE

    Szewczyk, B; Taylor, A

    1983-01-01

    An acidic polysaccharide of Escherichia coli B was isolated by a mild procedure and purified to homogeneity. The polysaccharide was found to react in Salmonella typhi Vi antisera and E. coli K1 antisera. Serological analysis and preliminary chemical characterization of the polysaccharide indicated that it is an aminouronic acid polymer which, although not structurally identical to either Vi or K1, appears more like the Vi antigen, both immunochemically and chemically.

  11. Mode of Action Temu Kunci (Kaempferia pandurata Essential Oil on E. coli K1.1 Cell Determined by Leakage of Material Cell and Salt Tolerance Assays

    Directory of Open Access Journals (Sweden)

    MIKSUSANTI

    2008-06-01

    Full Text Available The essential oil of Kaempferia pandurata consist of terpen and oxygenated terpen that exhibits broad-spectrum antimicrobial activity. It’s mode of action against the gram-negative bacterium E. coli K1.1 has been investigated using a range of treatments. The mode action of the essential oil were analyzed by it’s ability to leakage E. coli K1.1 cell, to change permeability of the cell, and to alter salt tolerance of the cell. Ion leakage from the cell were analyzed by atomic absorption spectrophotometer. Salt tolerance assays was conducted by investigating the ability of E. coli K1.1 treated with temu kunci essential oil to grow on NA supplemented with NaCl. Protein and acid nucleic leakage were analyzed by UV spectrophotometer. There were inorganic compound leakage (potassium, calcium ion and organic compound leakage (nucleic acid, protein from cytoplasmic membrane, after exposing this organism to essential oil of Kaempferia pandurata. The more concentration of oil added, the more leakage was observed due to the loss of absorbing material such as nucleic acid (260 nm and protein (280 nm, the loss of potassium and calcium ion, and loss of the salt tolerance of E. coli K1.1.

  12. The microbiota regulates neutrophil homeostasis and host resistance to Escherichia coli K1 sepsis in neonatal mice.

    Science.gov (United States)

    Deshmukh, Hitesh S; Liu, Yuhong; Menkiti, Ogechukwu R; Mei, Junjie; Dai, Ning; O'Leary, Claire E; Oliver, Paula M; Kolls, Jay K; Weiser, Jeffrey N; Worthen, G Scott

    2014-05-01

    Neonatal colonization by microbes, which begins immediately after birth, is influenced by gestational age and the mother's microbiota and is modified by exposure to antibiotics. In neonates, prolonged duration of antibiotic therapy is associated with increased risk of late-onset sepsis (LOS), a disorder controlled by neutrophils. A role for the microbiota in regulating neutrophil development and susceptibility to sepsis in the neonate remains unclear. We exposed pregnant mouse dams to antibiotics in drinking water to limit transfer of maternal microbes to the neonates. Antibiotic exposure of dams decreased the total number and composition of microbes in the intestine of the neonates. This was associated with decreased numbers of circulating and bone marrow neutrophils and granulocyte/macrophage-restricted progenitor cells in the bone marrow of antibiotic-treated and germ-free neonates. Antibiotic exposure of dams reduced the number of interleukin-17 (IL-17)-producing cells in the intestine and production of granulocyte colony-stimulating factor (G-CSF). Granulocytopenia was associated with impaired host defense and increased susceptibility to Escherichia coli K1 and Klebsiella pneumoniae sepsis in antibiotic-treated neonates, which could be partially reversed by administration of G-CSF. Transfer of a normal microbiota into antibiotic-treated neonates induced IL-17 production by group 3 innate lymphoid cells (ILCs) in the intestine, increasing plasma G-CSF levels and neutrophil numbers in a Toll-like receptor 4 (TLR4)- and myeloid differentiation factor 88 (MyD88)-dependent manner and restored IL-17-dependent resistance to sepsis. Specific depletion of ILCs prevented IL-17- and G-CSF-dependent granulocytosis and resistance to sepsis. These data support a role for the intestinal microbiota in regulation of granulocytosis, neutrophil homeostasis and host resistance to sepsis in neonates.

  13. 大肠杆菌44277(02:K1)中rmlB基因功能%Function of rmlB in the pathogenic Escherichia coli 44277( O2: k1)

    Institute of Scientific and Technical Information of China (English)

    丁卫平; 罗元明; 艾国民; 金城

    2011-01-01

    [ Objective ] To identify the role of rmlB in synthesizing L-rhamnose in the pathogenic Escherichia coli 44277 (O2:K1:H4). [Methods] The rmlB gene was expressed and the activity of the recombinant protein was assayed by measuring the quantity of reaction product. The rmlB gene was deleted by homologous recombination, then phenotypic changes of the △rmlB mutant was analyzed by Electron Microscope, Tricine SDS-PAGE and immunological methods. Further, various methods including MALDI-TOF-MS/MS, GC-MS and NMR was used to investigate the 0 antigen structure of the ArmlB mutant. [Results] RmlB was confirmed to be a protein harboring the activity of dTDP-D-glucose 4,6-dehydratase through enzyme assay. The ArmlB mutant was successfully constructed and no phenotypic change was observed after compared with the wild type strain. L-rhamnose still existed in the ArmlB mutant, indicating that there may be isoenzyme of RmlB presenting in the mutant or there was a novel way synthesizing L-rhamnose in the mutant. [Conclusion] RmlB has the activity of dTDP-D-glucose 4,6-dehydratase but it is not essential for the synthesis of L-rhamnose.%[目的]确定rmlB基因在大肠杆菌(02:K1)L-型鼠李糖合成中的作用.[方法]将基因rmlB进行原核表达并测定酶活;用同源重组的方法将rmlB基因敲除,分析表型变化,并运用质谱,以及核磁共振等手段分析脂多糖0侧链的结构,以确定rmlB在O抗原合成中的作用.[结果]成功对rmlB基因进行了表达并测定了重组蛋白的酶活,确定蛋白RmlB具有dTDP-D-glucose 4,6-dehydratase活性.成功构建了rmlB基因缺失突变株,对突变株进行表型分析发现突变株的表型与野生株相比无变化.对突变株分析发现突变株中的O抗原仍含有L-型鼠李糖,说明在该菌株中可能存在RmlB的同功能酶或者存在其它的L-型鼠李糖合成途径.[结论]rmlB基因编码的蛋白具有dTDP-D-glucose 4,6-dehydratase活性但此基因对于L-型鼠李糖的合成不是必需的.

  14. Interactions of Neuropathogenic Escherichia coli K1 (RS218 and Its Derivatives Lacking Genomic Islands with Phagocytic Acanthamoeba castellanii and Nonphagocytic Brain Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Farzana Abubakar Yousuf

    2014-01-01

    Full Text Available Here we determined the role of various genomic islands in E. coli K1 interactions with phagocytic A. castellanii and nonphagocytic brain microvascular endothelial cells. The findings revealed that the genomic islands deletion mutants of RS218 related to toxins (peptide toxin, α-hemolysin, adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (IbeA, CNF1, metabolism (D-serine catabolism, dihydroxyacetone, glycerol, and glyoxylate metabolism showed reduced interactions with both A. castellanii and brain microvascular endothelial cells. Interestingly, the deletion of RS218-derived genomic island 21 containing adhesins (P fimbriae, F17-like fimbriae, nonfimbrial adhesins, Hek, and hemagglutinin, protein secretion system (T1SS for hemolysin, invasins (CNF1, metabolism (D-serine catabolism abolished E. coli K1-mediated HBMEC cytotoxicity in a CNF1-independent manner. Therefore, the characterization of these genomic islands should reveal mechanisms of evolutionary gain for E. coli K1 pathogenicity.

  15. Meningitic Escherichia coli K1 penetration and neutrophil transmigration across the blood-brain barrier are modulated by alpha7 nicotinic receptor.

    Directory of Open Access Journals (Sweden)

    Feng Chi

    Full Text Available Alpha7 nicotinic acetylcholine receptor (nAChR, an essential regulator of inflammation, is abundantly expressed in hippocampal neurons, which are vulnerable to bacterial meningitis. However, it is unknown whether α7 nAChR contributes to the regulation of these events. In this report, an aggravating role of α7 nAChR in host defense against meningitic E. coli infection was demonstrated by using α7-deficient (α7(-/- mouse brain microvascular endothelial cells (BMEC and animal model systems. As shown in our in vitro and in vivo studies, E. coli K1 invasion and polymorphonuclear neutrophil (PMN transmigration across the blood-brain barrier (BBB were significantly reduced in α7(-/- BMEC and α7(-/- mice. Stimulation by nicotine was abolished in the α7(-/- cells and animals. The same blocking effect was achieved by methyllycaconitine (α7 antagonist. The tight junction molecules occludin and ZO-1 were significantly reduced in the brain cortex of wildtype mice infected with E. coli and treated with nicotine, compared to α7(-/- cells and animals. Decreased neuronal injury in the hippocampal dentate gyrus was observed in α7(-/- mice with meningitis. Proinflammatory cytokines (IL-1β, IL-6, TNFα, MCP-1, MIP-1alpha, and RANTES and adhesion molecules (CD44 and ICAM-1 were significantly reduced in the cerebrospinal fluids of the α7(-/- mice with E. coli meningitis. Furthermore, α7 nAChR is the major calcium channel for nicotine- and E. coli K1-increased intracellular calcium concentrations of mouse BMEC. Taken together, our data suggest that α7 nAChR plays a detrimental role in the host defense against meningitic infection by modulation of pathogen invasion, PMN recruitment, calcium signaling and neuronal inflammation.

  16. Protective effects of indigenous Escherichia coli against a pathogenic E. coli challenge strain in pigs.

    Science.gov (United States)

    Vahjen, W; Cuisiniere, T; Zentek, J

    2017-10-03

    To investigate the inhibitory effect of indigenous enterobacteria on pathogenic Escherichia coli, a challenge trial with postweaning pigs was conducted. A pathogenic E. coli strain was administered to all animals and their health was closely monitored thereafter. Faecal samples were taken from three healthy and three diarrhoeic animals. Samples were cultivated on MacConkey agar and isolates were subcultured. A soft agar overlay assay was used to determine the inhibitory activity of the isolates. A total of 1,173 enterobacterial isolates were screened for their ability to inhibit the E. coli challenge strain. Colony forming units of enterobacteria on MacConkey agar were not different between healthy and diarrhoeic animals in the original samples. Furthermore, numbers of isolates per animal were also not significantly different between healthy (482 isolates) and diarrhoeic animals (691 isolates). A total of 43 isolates (3.7%) with inhibitory activity against the pathogenic E. coli challenge strain were detected. All inhibitory isolates were identified as E. coli via MALDI-TOF. The isolates belonged to the phylotypes A, C and E. Many isolates (67.4%) were commensal E. coli without relevant porcine pathogenic factors, but toxin- and fimbrial genes (stx2e, fae, estIb, elt1a, fas, fan) were detected in 14 inhibitory isolates. Healthy animals showed significantly (P=0.003) more inhibitory isolates (36 of 482 isolates; 7.5%) than diseased animals (7 of 691 isolates; 1.0%). There were no significant correlations regarding phylotype or pathogenic factors between healthy and diseased animals. This study has shown that a small proportion of indigenous E. coli is able to inhibit in vitro growth of a pathogenic E. coli strain in pigs. Furthermore, healthy animals possess significantly more inhibitory E. coli strains than diarrhoeic animals. The inhibition of pathogenic E. coli by specific indigenous E. coli strains may be an underlying principle for the containment of pathogenic

  17. [Drug resistance of Escherichia coli strains isolated from poultry].

    Science.gov (United States)

    Giurov, B; Korudzhiĭski, N; Bineva, I

    1981-01-01

    Studied was the sensitivity of a total of 143 strains of Escherichia coli, isolated from young birds and broilers died from coli septicaemia, to antibiotics and chemotherapeutics. The following descending order was established: gentamycin, carbenicillin, ampicillin, furazolidon, borgal, kanamycin, strep tomycin, chloramphenicol, neomycin sulphathiazole, and tetracycline. Markers of resistance were established with all strains with regard to the therapeutic agents in current and prospective use in industrial poultry farming. It is stated that a preliminary antibiogram is indispensable in order to obtain dependable results in the treatment of animals affected with colibacteriosis. An alternative is to apply directly those drugs to which the strains have shown highest sensitivity.

  18. Gentamicin resistance among Escherichia coli strains isolated in neonatal sepsis.

    Science.gov (United States)

    Hasvold, J; Bradford, L; Nelson, C; Harrison, C; Attar, M; Stillwell, T

    2013-01-01

    Neonatal sepsis is a significant cause of morbidity and mortality among term and preterm infants. Ampicillin and gentamicin are standard empiric therapy for early onset sepsis. Four cases of neonatal sepsis secondary to Escherichia coli (E. coli) found to be gentamicin resistant occurred within a five week period in one neonatal intensive care unit (NICU). To determine whether these cases could be tied to a single vector of transmission, and to more broadly evaluate the incidence of gentamicin resistant strains of E. coli in the neonatal population at our institution compared to other centers, we reviewed the charts of the four neonates (Infants A through D) and their mothers. The E. coli isolates were sent for Pulse Field Gel Electrophoresis (PFGE) to evaluate for genetic similarity between strains. We also reviewed all positive E. coli cultures from one NICU over a two year period. Infants A and B had genetically indistinguishable strains which matched that of urine and placental cultures of Infant B's mother. Infant C had a genetically distinct organism. Infant D, the identical twin of Infant C, did not have typing performed. Review of all cultures positive for E. coli at our institution showed a 12.9 percent incidence of gentamicin-resistance. A review of other studies showed that rates of resistance vary considerably by institution. We conclude that gentamicin-resistant E. coli is a relatively uncommon cause of neonatal sepsis, but should remain a consideration in patients who deteriorate despite initiation of empiric antibiotics.

  19. Identification of diarrheagenic Escherichia coli strains from avian organic fertilizers.

    Science.gov (United States)

    Puño-Sarmiento, Juan; Gazal, Luis Eduardo; Medeiros, Leonardo P; Nishio, Erick K; Kobayashi, Renata K T; Nakazato, Gerson

    2014-08-28

    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.

  20. Identification of Diarrheagenic Escherichia coli Strains from Avian Organic Fertilizers

    Directory of Open Access Journals (Sweden)

    Juan Puño-Sarmiento

    2014-08-01

    Full Text Available 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.

  1. Analysis of the genetic determinants coding for the S-fimbrial adhesin (sfa) in different Escherichia coli strains causing meningitis or urinary tract infections.

    Science.gov (United States)

    Ott, M; Hacker, J; Schmoll, T; Jarchau, T; Korhonen, T K; Goebel, W

    1986-12-01

    Recently we have described the molecular cloning of the genetic determinant coding for the S-fimbrial adhesin (Sfa), a sialic acid-recognizing pilus frequently found among extraintestinal Escherichia coli isolates. Fimbriae from the resulting Sfa+ E. coli K-12 clone were isolated, and an Sfa-specific antiserum was prepared. Western blots indicate that S fimbriae isolated from different uropathogenic and meningitis-associated E. coli strains, including O83:K1 isolates, were serologically related. The Sfa-specific antibodies did not cross-react with P fimbriae, but did cross-react with F1C fimbriae. Furthermore the sfa+ recombinant DNAs and some cloned sfa-flanking regions were used as probes in Southern experiments. Chromosomal DNAs isolated from O18:K1 and O83:K1 meningitis strains with and without S fimbriae and from uropathogenic O6:K+ strains were hybridized against these sfa-specific probes. Only one copy of the sfa determinant was identified on the chromosome of these strains. No sfa-specific sequences were observed on the chromosome of E. coli K-12 strains and an O7:K1 isolate. With the exception of small alterations in the sfa-coding region the genetic determinants for S fimbriae were identical in uropathogenic O6:K+ and meningitis O18:K1 and O83:K1 strains. The sfa determinant was also detected on the chromosome of K1 isolates with an Sfa-negative phenotype, and specific cross-hybridization signals were visible after blotting against F1C-specific DNA. In addition homology among the different strains was observed in the sfa-flanking regions.

  2. Phylogenetic analysis of Escherichia coli strains isolated from human samples

    Directory of Open Access Journals (Sweden)

    Abdollah Derakhshandeh

    2013-12-01

    Full Text Available Escherichia coli (E. coli is a normal inhabitant of the gastrointestinal tract of vertebrates, including humans. Phylogenetic analysis has shown that E. coli is composed of four main phylogenetic groups (A, B1, B2 and D. Group A and B1 are generally associated with commensals, whereas group B2 is associated with extra-intestinal pathotypes. Most enteropathogenic isolates, however, are assigned to group D. In the present study, a total of 102 E. coli strains, isolated from human samples, were used. Phylogenetic grouping was done based on the Clermont triplex PCR method using primers targeted at three genetic markers, chuA, yjaA and TspE4.C2. Group A contained the majority of the collected isolates (69 isolates, 67.64%, followed by group B2 (18 isolates, 17.64% and D (15 isolates, 14.7% and no strains were found to belong to group B1. The distribution of phylogenetic groups in our study suggests that although the majority of strains were commensals, the prevalence of enteropathogenic and extra-intestinal pathotypes was noteworthy. Therefore, the role of E. coli in human infections including diarrhea, urinary tract infections and meningitis should be considered.

  3. Characterization of urinary Escherichia coli O75 strains.

    Science.gov (United States)

    Nimmich, W; Voigt, W; Seltmann, G

    1997-01-01

    Forty-four Escherichia coli O75 strains from patients with urinary tract infections were characterized by a variety of methods to obtain evidence of their clonal distribution and uropathogenic properties. By K and H antigen typing, the strains were divided into the following serotypes: O75:K5:H- (18 strains), O75:K95:H- (10 strains), O75:K95:H5 (7 strains), O75:K100:H5 (4 strains), and O75:K-:H55 (5 strains). Generally, biotyping proved to be of no discriminative value. With two exceptions the strains were found to be sensitive to the bactericidal effect of normal human serum. As shown by multilocus enzyme electrophoresis, the whole-cell protein profile (WCPP), and the patterns of the outer membrane proteins and lipopolysaccharides, all but the five O75:H55 strains were genetically closely related to each other and could be classified into one clonal group. The O75:K-:H55 strains proved to be quite different and lacked type 1 fimbriae. All 17 K95 (H-, H5) strains produced hemolysin and P fimbriae. Five of the O75:K5:H- strains were different from the other K5 strains by showing hemagglutinating properties, on the basis of the presence of the OX adhesin. The last two groups are suggested to be uropathogenic and are proposed to represent separate clonal groups or subgroups. PMID:9114391

  4. [Investigation of pathogenic Escherichia coli strains in patients with diarrhea].

    Science.gov (United States)

    Aydın Tutak, Gülten; Tuğrul, Hamdi Murat

    2015-01-01

    The role of certain serogroups and serotypes of Escherichia coli in the etiology of gastroenteritis is increasingly appreciated. It is important to detect the virulence factors of diarrheagenic E.coli strains that differentiate them from nonpathogenic members of normal intestinal flora for the diagnosis and treatment. The aims of this study were to determine the serotypes of E.coli isolates that cause gastroenteritis and to investigate the presence of virulence genes by polymerase chain reaction (PCR). A total of 202 watery, bloody or mucoid stool samples sent to microbiology laboratory collected from patients with diarrhea who were admitted to outpatient clinics of Trakya University Health Research and Application Hospital between February to October 2009, were included in the study. A total of 254 predominantly grown E.coli strains have been isolated and identified with conventional methods from the cultures of those 202 samples. All strains were tested by slide agglutination (SA) that includes 6 units of O serogroups polyvalent antisera of enteropathogenic E.coli (EPEC), enterotoxigenic E.coli (ETEC) and enteroinvasive E.coli (EIEC). The samples which yielded positive results with SA test and the same number of negative samples selected with mapping method as controls were studied for the presence of virulence genes belonging EPEC, ETEC and EIEC by conventional PCR. In the study, 14.3% (29/202) of the samples were serogrouped with SA, of them 13 (6.4%) were identified as EPEC, 11 (5.4%) as EIEC and five (2.4%) as ETEC. Only five isolates belonging to EPEC serogroup could be defined by monovalent antiserum and they were all in O1 serogroup. Out of 29 pathogenic E.coli serotyped, 3 (10.3%) of them harbored the virulence genes of diarrheagenic strains. One sample which was positive for eaeA gene of EPEC, did not harbor bfpA and stx genes and was defined as atypical EPEC. Out of other two samples, one was positive for estA gene of ETEC and the other one for ial gene

  5. Comparative Genomics of Escherichia coli Strains Causing Urinary Tract Infections

    DEFF Research Database (Denmark)

    Vejborg, Rebecca Munk; Hancock, Viktoria; Schembri, Mark A.

    2011-01-01

    The virulence determinants of uropathogenic Escherichia coli have been studied extensively over the years, but relatively little is known about what differentiates isolates causing various types of urinary tract infections. In this study, we compared the genomic profiles of 45 strains from a range...

  6. Molecular characterization of the Escherichia coli asymptomatic bacteriuria strain 83972

    DEFF Research Database (Denmark)

    Klemm, Per; Hancock, Viktoria; Ulett, G.C.

    2006-01-01

    Escherichia coli 83972 is a clinical asymptomatia bacteriuric isolate that is able to colonize the human urinary bladder without inducing an immune response. Here we demonstrate that one of the mechanisms by which this strain has become attenuated is through the mutation of its genes encoding type...

  7. Molecular characterization of the Escherichia coli asymptomatic bacteriuria strain 83972

    DEFF Research Database (Denmark)

    Klemm, Per; Hancock, Viktoria; Ulett, G.C.

    2006-01-01

    Escherichia coli 83972 is a clinical asymptomatia bacteriuric isolate that is able to colonize the human urinary bladder without inducing an immune response. Here we demonstrate that one of the mechanisms by which this strain has become attenuated is through the mutation of its genes encoding typ...

  8. Collagen-like proteins in pathogenic E. coli strains.

    Directory of Open Access Journals (Sweden)

    Neelanjana Ghosh

    Full Text Available The genome sequences of enterohaemorrhagic E. coli O157:H7 strains show multiple open-reading frames with collagen-like sequences that are absent from the common laboratory strain K-12. These putative collagens are included in prophages embedded in O157:H7 genomes. These prophages carry numerous genes related to strain virulence and have been shown to be inducible and capable of disseminating virulence factors by horizontal gene transfer. We have cloned two collagen-like proteins from E. coli O157:H7 into a laboratory strain and analysed the structure and conformation of the recombinant proteins and several of their constituting domains by a variety of spectroscopic, biophysical, and electron microscopy techniques. We show that these molecules exhibit many of the characteristics of vertebrate collagens, including trimer formation and the presence of a collagen triple helical domain. They also contain a C-terminal trimerization domain, and a trimeric α-helical coiled-coil domain with an unusual amino acid sequence almost completely lacking leucine, valine or isoleucine residues. Intriguingly, these molecules show high thermal stability, with the collagen domain being more stable than those of vertebrate fibrillar collagens, which are much longer and post-translationally modified. Under the electron microscope, collagen-like proteins from E. coli O157:H7 show a dumbbell shape, with two globular domains joined by a hinged stalk. This morphology is consistent with their likely role as trimeric phage side-tail proteins that participate in the attachment of phage particles to E. coli target cells, either directly or through assembly with other phage tail proteins. Thus, collagen-like proteins in enterohaemorrhagic E. coli genomes may have a direct role in the dissemination of virulence-related genes through infection of harmless strains by induced bacteriophages.

  9. Diarrhea, Urosepsis and Hemolytic Uremic Syndrome Caused by the Same Heteropathogenic Escherichia coli Strain.

    Science.gov (United States)

    Ang, C Wim; Bouts, Antonia H M; Rossen, John W A; Van der Kuip, Martijn; Van Heerde, Marc; Bökenkamp, Arend

    2016-09-01

    We describe an 8-month-old girl with diarrhea, urosepsis and hemolytic uremic syndrome caused by Escherichia coli. Typing of cultured E. coli strains from urine and blood revealed the presence of virulence factors from multiple pathotypes of E. coli. This case exemplifies the genome plasticity of E. coli and the resulting heteropathogenic strains.

  10. Uropathogenic Escherichia coli (UPEC) strains may carry virulence properties of diarrhoeagenic E. coli.

    Science.gov (United States)

    Abe, Cecilia M; Salvador, Fábia A; Falsetti, Ivan N; Vieira, Mônica A M; Blanco, Jorge; Blanco, Jesús E; Blanco, Miguel; Machado, Antônia M O; Elias, Waldir P; Hernandes, Rodrigo T; Gomes, Tânia A T

    2008-04-01

    To analyze whether Escherichia coli strains that cause urinary tract infections (UPEC) share virulence characteristics with the diarrheagenic E. coli (DEC) pathotypes and to recognize their genetic diversity, 225 UPEC strains were examined for the presence of various properties of DEC and UPEC (type of interaction with HeLa cells, serogroups and presence of 30 virulence genes). No correlation between adherence patterns and serogroups was observed. Forty-five serogroups were found, but 64% of the strains belonged to one of the 12 serogroups (O1, O2, O4, O6, O7, O14, O15, O18, O21, O25, O75, and O175) and carried UPEC virulence genes (pap, hly, aer, sfa, cnf). The DEC genes found were: aap, aatA, aggC, agg3C, aggR, astA, eae, ehly, iha, irp2, lpfA(O113), pet, pic, pilS, and shf. Sixteen strains presented aggregative adherence and/or the aatA sequence, which are characteristics of enteroaggregative E. coli (EAEC), one of the DEC pathotypes. In summary, certain UPEC strains may carry DEC virulence properties, mostly associated to the EAEC pathotype. This finding raises the possibility that at least some faecal EAEC strains might represent potential uropathogens. Alternatively, certain UPEC strains may have acquired EAEC properties, becoming a potential cause of diarrhoea.

  11. Transcriptomics and adaptive genomics of the asymptomatic bacteriuria Escherichia coli strain 83972

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Seshasayee, Aswin S.; Ussery, David

    2008-01-01

    Escherichia coli strains are the major cause of urinary tract infections in humans. Such strains can be divided into virulent, UPEC strains causing symptomatic infections, and asymptomatic, commensal-like strains causing asymptomatic bacteriuria, ABU. The best-characterized ABU strain is strain....... Strain 83972 is a deconstructed pathogen rather than a commensal strain that has acquired fitness properties....

  12. Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells

    Science.gov (United States)

    Starost, Laura Julia; Karassek, Sascha; Sano, Yasuteru; Kanda, Takashi; Kim, Kwang Sik; Dobrindt, Ulrich; Rüter, Christian; Schmidt, Marcus Alexander

    2016-01-01

    Pertussis toxin (PTx), the major virulence factor of the whooping cough-causing bacterial pathogen Bordetella pertussis, permeabilizes the blood–brain barrier (BBB) in vitro and in vivo. Breaking barriers might promote translocation of meningitis-causing bacteria across the BBB, thereby facilitating infection. PTx activates several host cell signaling pathways exploited by the neonatal meningitis-causing Escherichia coli K1-RS218 for invasion and translocation across the BBB. Here, we investigated whether PTx and E. coli K1-RS218 exert similar effects on MAPK p38, NF-κB activation and transcription of downstream targets in human cerebral endothelial TY10 cells using qRT-PCR, Western blotting, and ELISA in combination with specific inhibitors. PTx and E. coli K1-RS218 activate MAPK p38, but only E. coli K1-RS218 activates the NF-κB pathway. mRNA and protein levels of p38 and NF-κB downstream targets including IL-6, IL-8, CxCL-1, CxCL-2 and ICAM-1 were increased. The p38 specific inhibitor SB203590 blocked PTx-enhanced activity, whereas E. coli K1-RS218’s effects were inhibited by the NF-κB inhibitor Bay 11-7082. Further, we found that PTx enhances the adherence of human monocytic THP-1 cells to human cerebral endothelial TY10 cells, thereby contributing to enhanced translocation. These modulations of host cell signaling pathways by PTx and meningitis-causing E. coli support their contributions to pathogen and monocytic THP-1 cells translocation across the BBB. PMID:27754355

  13. Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells.

    Science.gov (United States)

    Starost, Laura Julia; Karassek, Sascha; Sano, Yasuteru; Kanda, Takashi; Kim, Kwang Sik; Dobrindt, Ulrich; Rüter, Christian; Schmidt, Marcus Alexander

    2016-10-13

    Pertussis toxin (PTx), the major virulence factor of the whooping cough-causing bacterial pathogen Bordetella pertussis, permeabilizes the blood-brain barrier (BBB) in vitro and in vivo. Breaking barriers might promote translocation of meningitis-causing bacteria across the BBB, thereby facilitating infection. PTx activates several host cell signaling pathways exploited by the neonatal meningitis-causing Escherichia coli K1-RS218 for invasion and translocation across the BBB. Here, we investigated whether PTx and E. coli K1-RS218 exert similar effects on MAPK p38, NF-κB activation and transcription of downstream targets in human cerebral endothelial TY10 cells using qRT-PCR, Western blotting, and ELISA in combination with specific inhibitors. PTx and E. coli K1-RS218 activate MAPK p38, but only E. coli K1-RS218 activates the NF-κB pathway. mRNA and protein levels of p38 and NF-κB downstream targets including IL-6, IL-8, CxCL-1, CxCL-2 and ICAM-1 were increased. The p38 specific inhibitor SB203590 blocked PTx-enhanced activity, whereas E. coli K1-RS218's effects were inhibited by the NF-κB inhibitor Bay 11-7082. Further, we found that PTx enhances the adherence of human monocytic THP-1 cells to human cerebral endothelial TY10 cells, thereby contributing to enhanced translocation. These modulations of host cell signaling pathways by PTx and meningitis-causing E. coli support their contributions to pathogen and monocytic THP-1 cells translocation across the BBB.

  14. Pertussis Toxin Exploits Host Cell Signaling Pathways Induced by Meningitis-Causing E. coli K1-RS218 and Enhances Adherence of Monocytic THP-1 Cells to Human Cerebral Endothelial Cells

    Directory of Open Access Journals (Sweden)

    Laura Julia Starost

    2016-10-01

    Full Text Available Pertussis toxin (PTx, the major virulence factor of the whooping cough-causing bacterial pathogen Bordetella pertussis, permeabilizes the blood–brain barrier (BBB in vitro and in vivo. Breaking barriers might promote translocation of meningitis-causing bacteria across the BBB, thereby facilitating infection. PTx activates several host cell signaling pathways exploited by the neonatal meningitis-causing Escherichia coli K1-RS218 for invasion and translocation across the BBB. Here, we investigated whether PTx and E. coli K1-RS218 exert similar effects on MAPK p38, NF-κB activation and transcription of downstream targets in human cerebral endothelial TY10 cells using qRT-PCR, Western blotting, and ELISA in combination with specific inhibitors. PTx and E. coli K1-RS218 activate MAPK p38, but only E. coli K1-RS218 activates the NF-κB pathway. mRNA and protein levels of p38 and NF-κB downstream targets including IL-6, IL-8, CxCL-1, CxCL-2 and ICAM-1 were increased. The p38 specific inhibitor SB203590 blocked PTx-enhanced activity, whereas E. coli K1-RS218’s effects were inhibited by the NF-κB inhibitor Bay 11-7082. Further, we found that PTx enhances the adherence of human monocytic THP-1 cells to human cerebral endothelial TY10 cells, thereby contributing to enhanced translocation. These modulations of host cell signaling pathways by PTx and meningitis-causing E. coli support their contributions to pathogen and monocytic THP-1 cells translocation across the BBB.

  15. Examination of uropathogenic Escherichia coli strains conferring large plasmids

    Directory of Open Access Journals (Sweden)

    SUHARTONO

    2010-04-01

    Full Text Available Suhartono (2010 Examination of uropathogenic Escherichia coli strains conferring large plasmids. Biodiversitas 11: 59-64. Of major uropathogens, Escherichia coli has been widely known as a main pathogen of UTIs globally and has considerable medical and financial consequences. A strain of UPEC, namely E. coli ST131, confers a large plasmid encoding cephalosporinases (class C β-lactamase or AmpC that may be disseminated through horizontal transfer among bacterial populations. Therefore, it is worth examining such large plasmids by isolating, purifying, and digesting the plasmid with restriction enzymes. The examination of the large plasmids was conducted by isolating plasmid DNA visualized by agarose gel electrophoresis as well as by PFGE. The relationship of plasmids among isolates was carried out by HpaI restriction enzyme digestion. Of 36 isolates of E. coli ST 131, eight isolates possessed large plasmids, namely isolates 3, 9, 10, 12, 17, 18, 26 and 30 with the largest molecular size confirmed by agarose gel electrophoresis and PFGE was ~42kb and ~118kb respectively. Restriction enzyme analysis revealed that isolates 9, 10, 12, 17 and 18 have the common restriction patterns and those isolates might be closely related.

  16. Unusual "flesh-eating" strains of Escherichia coli.

    Science.gov (United States)

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

    2012-12-01

    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.

  17. Impact of Diversity of Colonizing Strains on Strategies for Sampling Escherichia coli from Fecal Specimens ▿

    Science.gov (United States)

    Lautenbach, Ebbing; Bilker, Warren B.; Tolomeo, Pam; Maslow, Joel N.

    2008-01-01

    Of 49 subjects, 21 were colonized with more than one strain of Escherichia coli and 12 subjects had at least one strain present in fewer than 20% of colonies. The ability to accurately characterize E. coli strain diversity is directly related to the number of colonies sampled and the underlying prevalence of the strain. PMID:18650357

  18. Impact of diversity of colonizing strains on strategies for sampling Escherichia coli from fecal specimens.

    Science.gov (United States)

    Lautenbach, Ebbing; Bilker, Warren B; Tolomeo, Pam; Maslow, Joel N

    2008-09-01

    Of 49 subjects, 21 were colonized with more than one strain of Escherichia coli and 12 subjects had at least one strain present in fewer than 20% of colonies. The ability to accurately characterize E. coli strain diversity is directly related to the number of colonies sampled and the underlying prevalence of the strain.

  19. Inactivation and Gene Expression of a Virulent Wastewater Escherichia coli Strain and the Nonvirulent Commensal Escherichia coli DSM1103 Strain upon Solar Irradiation

    KAUST Repository

    Aljassim, Nada I.

    2017-03-06

    This study examined the decay kinetics and molecular responses of two Escherichia coli strains upon solar irradiation. The first is E. coli PI-7, a virulent and antibiotic-resistant strain that was isolated from wastewater and carries the emerging NDM-1 antibiotic resistance gene. The other strain, E. coli DSM1103, displayed lower virulence and antibiotic resistance than E. coli PI-7. In a buffer solution, E. coli PI-7 displayed a longer lag phase prior to decay and a longer half-life compared with E. coli DSM1103 (6.64 ± 0.63 h and 2.85 ± 0.46 min vs 1.33 ± 0.52 h and 2.04 ± 0.36 min). In wastewater, both E. coli strains decayed slower than they did in buffer. Although solar irradiation remained effective in reducing the numbers of both strains by more than 5-log10 in <24 h, comparative genomics and transcriptomics revealed differences in the genomes and overall regulation of genes between the two E. coli strains. A wider arsenal of genes related to oxidative stress, cellular repair and protective mechanisms were upregulated in E. coli PI-7. Subpopulations of E. coli PI-7 expressed genes related to dormancy and persister cell formation during the late decay phase, which may have accounted for its prolonged persistence. Upon prolonged solar irradiation, both E. coli strains displayed upregulation of genes related to horizontal gene transfer and antibiotic resistance. Virulence functions unique to E. coli PI-7 were also upregulated. Our findings collectively indicated that, whereas solar irradiation is able to reduce total cell numbers, viable E. coli remained and expressed genes that enable survival despite solar treatment. There remains a need for heightened levels of concern regarding risks arising from the dissemination of E. coli that may remain viable in wastewater after solar irradiation.

  20. Coordination-resolved local bond strain and 3p energy entrapment of K atomic clusters and K(1 1 0) skin

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Ting; Bo, Maolin; Guo, Yongling [Key Laboratory of Low-Dimensional Materials and Application Technologies (Ministry of Education), Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Hunan Provincial Key Laboratory of Thin Film Materials and Devices, Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Chen, Hefeng [United Superconductive Institution, Shanghai Jiaotong University, Shanghai 200240 (China); Wang, Yan [School of Information and Electronic Engineering, Hunan University of Science and Technology, Hunan 411201 (China); Huang, Yongli, E-mail: huangyongli@xtu.edu.cn [Key Laboratory of Low-Dimensional Materials and Application Technologies (Ministry of Education), Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Hunan Provincial Key Laboratory of Thin Film Materials and Devices, Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Sun, Chang Q., E-mail: ecqsun@ntu.edu.sg [Key Laboratory of Low-Dimensional Materials and Application Technologies (Ministry of Education), Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); Hunan Provincial Key Laboratory of Thin Film Materials and Devices, Faculty of Materials Science and Engineering, Xiangtan University, Hunan 411105 (China); School of Information and Electronic Engineering, Hunan University of Science and Technology, Hunan 411201 (China); NOVITAS, School of Electrical and Electronic Engineering, Nanyang Technological University, Singapore 639798 (Singapore)

    2015-09-15

    Graphical abstract: - Highlights: • Coordination environment resolves electron binding-energy shift of K{sub 44}, K{sub 46}, K{sub 55} clusters. • Predict the effective coordination number of K nanoclusters when we get the atomic number N. • Atomic under coordination shortens the local bonds and entrapment. • XPS derives core level of an isolated atom and its bulk shift. - Abstract: We have examined the atomic coordination effect on the local bond strain and the 3p core-level shift of K(1 1 0) skin and nanoclusters using a combination of the bond order–length–strength correlation notion, tight-binding approach, density functional theory calculations, and photoelectron spectroscopy measurements. It turns out that: (i) the 3p core-level shifts from 15.595 ± 0.003 eV for an isolated K atom by 2.758 eV to the bulk value of 18.353 eV; (ii) the effective atomic coordination number reduces from the bulk value of 12 to 3.93 for the first layer and to 5.81 for the second layer of K(1 1 0) skin associated with the local lattice strain of 12.76%, a binding energy density 72.67%, and atomic cohesive energy −62.46% for the skin; and (iii) K cluster size reduction lowers the effective atomic coordination number and enhances further the skin electronic attribution. Results have revealed that the 3p core-level shifts of K(1 1 0) and nanoclusters originate from perturbation of the Hamiltonian by under-coordination induced charge densification and quantum entrapment.

  1. Characterisation of atypical enteropathogenic E. coli strains of clinical origin

    Directory of Open Access Journals (Sweden)

    Tennant Sharon M

    2009-06-01

    Full Text Available Abstract Background Enteropathogenic E. coli (EPEC is a prominent cause of diarrhoea, and is characterised in part by its carriage of a pathogenicity island: the locus for enterocyte effacement (LEE. EPEC is divided into two subtypes according to the presence of bundle-forming pili (BFP, a fimbrial adhesin that is a virulence determinant of typical EPEC (tEPEC, but is absent from atypical EPEC (aEPEC. Because aEPEC lack BFP, their virulence has been questioned, as they may represent LEE-positive Shiga toxin-producing E. coli (STEC that have lost the toxin-encoding prophage, or tEPEC that have lost the genes for BFP. To determine if aEPEC isolated from humans in Australia or New Zealand fall into either of these categories, we undertook phylogenetic analysis of 75 aEPEC strains, and compared them with reference strains of EPEC and STEC. We also used PCR and DNA hybridisation to determine if aEPEC carry virulence determinants that could compensate for their lack of BFP. Results The results showed that aEPEC are highly heterogeneous. Multilocus sequence typing revealed that 61 of 75 aEPEC strains did not belong to known tEPEC or STEC clades, and of those that did, none expressed an O:H serotype that is frequent in tEPEC or STEC strains associated with disease. PCR for each of 18 known virulence-associated determinants of E. coli was positive in less than 15% of strains, apart from NleB which was detected in 30%. Type I fimbriae were expressed by all aEPEC strains, and 12 strains hybridised with DNA probes prepared from either bfpA or bfpB despite being negative in the PCR for bfpA. Conclusion Our findings indicate that clinical isolates of aEPEC obtained from patients in Australia or New Zealand are not derived from tEPEC or STEC, and suggest that functional equivalents of BFP and possibly type I fimbriae may contribute to the virulence of some aEPEC strains.

  2. Complete genome sequences of Escherichia coli strains 1303 and ECC-1470 isolated from bovine mastitis

    NARCIS (Netherlands)

    Leimbach, Andreas; Poehlein, Anja; Witten, Anika; Scheutz, Flemming; Schukken, Ynte; Daniel, Rolf; Dobrindt, Ulrich

    2016-01-01

    Escherichia coli is the leading causative agent of acute bovine mastitis. Here, we report the complete genome sequence of E. coli O70:H32 strain 1303, isolated from an acute case of bovine mastitis, and E. coli Ont:Hnt strain ECC-1470, isolated from a persistent infection.

  3. Synthesis, biological evaluation, and structure-activity relationships of N-benzoyl-2-hydroxybenzamides as agents active against P. falciparum (K1 strain), Trypanosomes, and Leishmania.

    Science.gov (United States)

    Stec, Jozef; Huang, Qingqing; Pieroni, Marco; Kaiser, Marcel; Fomovska, Alina; Mui, Ernest; Witola, William H; Bettis, Samuel; McLeod, Rima; Brun, Reto; Kozikowski, Alan P

    2012-04-12

    In our efforts to identify novel chemical scaffolds for the development of new antiprotozoal drugs, a compound library was screened against Toxoplasma gondii tachyzoites with activity discovered for N-(4-ethylbenzoyl)-2-hydroxybenzamide 1a against T. gondii as described elsewhere. Synthesis of a compound set was guided by T. gondii SAR with 1r found to be superior for T. gondii , also active against Thai and Sierra Leone strains of Plasmodium falciparum , and with superior ADMET properties as described elsewhere. Herein, synthesis methods and details of the chemical analysis of the compounds in this series are described. Further, this series of N-benzoyl-2-hydroxybenzamides was repurposed for testing against four other protozoan parasites: Trypanosoma brucei rhodesiense , Trypanosoma cruzi , Leishmania donovani , and P. falciparum (K1 isolate). Structure-activity analyses led to the identification of compounds in this set with excellent antileishmanial activity (compound 1d). Overall, compound 1r was the best and had activity 21-fold superior to that of the standard antimalarial drug chloroquine against the K1 P. falciparum isolate. © 2012 American Chemical Society

  4. Origins of the E. coli Strain Causing an Outbreak of Hemolytic–Uremic Syndrome in Germany

    Science.gov (United States)

    Rasko, David A.; Webster, Dale R.; Sahl, Jason W.; Bashir, Ali; Boisen, Nadia; Scheutz, Flemming; Paxinos, Ellen E.; Sebra, Robert; Chin, Chen-Shan; Iliopoulos, Dimitris; Klammer, Aaron; Peluso, Paul; Lee, Lawrence; Kislyuk, Andrey O.; Bullard, James; Kasarskis, Andrew; Wang, Susanna; Eid, John; Rank, David; Redman, Julia C.; Steyert, Susan R.; Frimodt-Møller, Jakob; Struve, Carsten; Petersen, Andreas M.; Krogfelt, Karen A.; Nataro, James P.; Schadt, Eric E.; Waldor, Matthew K.

    2011-01-01

    BACKGROUND A large outbreak of diarrhea and the hemolytic–uremic syndrome caused by an unusual serotype of Shiga-toxin–producing Escherichia coli (O104:H4) began in Germany in May 2011. As of July 22, a large number of cases of diarrhea caused by Shiga-toxin–producing E. coli have been reported — 3167 without the hemolytic–uremic syndrome (16 deaths) and 908 with the hemolytic–uremic syndrome (34 deaths) — indicating that this strain is notably more virulent than most of the Shiga-toxin–producing E. coli strains. Preliminary genetic characterization of the outbreak strain suggested that, unlike most of these strains, it should be classified within the enteroaggregative pathotype of E. coli. METHODS We used third-generation, single-molecule, real-time DNA sequencing to determine the complete genome sequence of the German outbreak strain, as well as the genome sequences of seven diarrhea-associated enteroaggregative E. coli serotype O104:H4 strains from Africa and four enteroaggregative E. coli reference strains belonging to other serotypes. Genomewide comparisons were performed with the use of these enteroaggregative E. coli genomes, as well as those of 40 previously sequenced E. coli isolates. RESULTS The enteroaggregative E. coli O104:H4 strains are closely related and form a distinct clade among E. coli and enteroaggregative E. coli strains. However, the genome of the German outbreak strain can be distinguished from those of other O104:H4 strains because it contains a prophage encoding Shiga toxin 2 and a distinct set of additional virulence and antibiotic-resistance factors. CONCLUSIONS Our findings suggest that horizontal genetic exchange allowed for the emergence of the highly virulent Shiga-toxin–producing enteroaggregative E. coli O104:H4 strain that caused the German outbreak. More broadly, these findings highlight the way in which the plasticity of bacterial genomes facilitates the emergence of new pathogens. PMID:21793740

  5. Emergence of serotype K1 Klebsiella pneumoniae ST23 strains co-producing the plasmid-mediated AmpC beta-lactamase DHA-1 and an extended-spectrum beta-lactamase in Korea

    Directory of Open Access Journals (Sweden)

    Hae Suk Cheong

    2016-11-01

    Full Text Available Abstract Background Serotype K1 Klebsiella pneumoniae has emerged as an important community pathogen causing various infections, including liver abscesses. Although serotype K1 K. pneumoniae community isolates have been reported as susceptible to most classes of antimicrobial agents, a few cases of infection caused by extended-spectrum beta-lactamase (ESBL-producing serotype K1 K. pneumoniae have recently been reported in Asian countries. We identified three ESBL-producing strains of serotype K1 K. pneumoniae and conducted a molecular characterization of their drug resistance. Methods Three ESBL-producing serotype K1 K. pneumoniae ST23 strains were identified from strains in the Asian Bacterial Bank. Antimicrobial susceptibility testing was performed using the broth microdilution method, and ESBL production was tested by the double-disk synergy test and a confirmatory test. PCR was performed to detect the genes for plasmid-mediated ESBL and AmpC beta-lactamases. Results All three strains were resistant to cefotaxime, ceftazidime, and piperacillin/tazobactam, and all were determined to be ESBL-producers. No known ESBL genes, including bla SHV, bla TEM, bla CTX-M, bla GES, bla PER, and bla VEB, were detected among the three strains. Of all plasmid-mediated AmpC beta-lactamase (PAB genes, including bla DHA-1, bla CMY, bla FOX, and bla MOX, the bla DHA-1 gene was detected in two of the strains. The PFGE patterns revealed that the two isolates carrying bla DHA-1 were closely related (84% similarity. Conclusions No ESBL genes were detected among three ESBL-producing serotype K1 K. pneumoniae ST23 strains. Two strains contained the PAB gene bla DHA-1. The emergence of resistant strains of community-origin serotype K1 K. pneumoniae has important implications for effective treatment and infection control practices.

  6. Short communication: heat resistance of Escherichia coli strains in raw milk at different subpasteurization conditions.

    Science.gov (United States)

    Peng, S; Hummerjohann, J; Stephan, R; Hammer, P

    2013-06-01

    A commonly applied treatment of raw milk to reduce bacterial loads is the short-time application of heat at subpasteurization levels under continuous flow, generally referred to as thermization, because this method retains some of the beneficial properties of raw milk. In a previous study, Escherichia coli strains exhibiting increased thermotolerance were found, demanding investigations into their ability to survive thermization. Nine E. coli strains, including 4 Shiga toxin-producing E. coli (STEC) strains, were investigated for their reduction during a thermization treatment in raw milk using a pilot-plant pasteurizer to reflect typically applied commercial conditions. Six of the 9 E. coli strains, including the 4 STEC strains, were similarly inactivated at 60, 62.5, and 65°C, whereas increased thermotolerance was observed for 3 E. coli strains. All strains were reduced to thermization treatment.

  7. In vitro evolution of an archetypal enteropathogenic Escherichia coli strain.

    Science.gov (United States)

    Nisa, Shahista; Hazen, Tracy H; Assatourian, Lillian; Nougayrède, Jean-Philippe; Rasko, David A; Donnenberg, Michael S

    2013-10-01

    Enteropathogenic Escherichia coli (EPEC) is a leading cause of infantile diarrhea in developing countries. EPEC strain E2348/69 is used worldwide as a prototype to study EPEC genetics and disease. However, isolates of E2348/69 differ phenotypically, reflecting a history of in vitro selection. To identify the genomic and phenotypic changes in the prototype strain, we sequenced the genome of the nalidixic acid-resistant (Nal(r)) E2348/69 clone. We also sequenced a recent nleF mutant derived by one-step PCR mutagenesis from the Nal(r) strain. The sequencing results revealed no unintended changes between the mutant and the parent strain. However, loss of the pE2348-2 plasmid and 3 nonsynonymous mutations were found in comparison to the published streptomycin-resistant (Str(r)) E2348/69 reference genome. One mutation is a conservative amino acid substitution in ftsK. Another, in gyrA, is a mutation known to result in resistance to nalidixic acid. The third mutation converts a stop codon to a tryptophan, predicted to result in the fusion of hflD, the lysogenization regulator, to purB. The purB gene encodes an adenylosuccinate lyase involved in purine biosynthesis. The Nal(r) clone has a lower growth rate than the Str(r) isolate when cultured in minimal media, a difference which is corrected upon addition of adenine or by genetic complementation with purB. Addition of adenine or genetic complementation also restored the invasion efficiency of the Nal(r) clone. This report reconciles longstanding inconsistencies in phenotypic properties of an archetypal strain and provides both reassurance and cautions regarding intentional and unintentional evolution in vitro.

  8. Sorbitol-negative phenotype among enterohemorrhagic Escherichia coli strains of different serotypes and from different sources.

    Science.gov (United States)

    Ojeda, A; Prado, V; Martinez, J; Arellano, C; Borczyk, A; Johnson, W; Lior, H; Levine, M M

    1995-08-01

    Enterohemorrhagic Escherichia coli (EHEC) strains detected with DNA probes (for virulence plasmid and Shiga-like toxins) from subjects with hemolytic-uremic syndrome (n = 19) or diarrhea (n = 41) or asymptomatic carriers (n = 29) were examined for sorbitol fermentability, as were enterotoxigenic (n = 40), enteropathogenic (n = 40), and enteroinvasive (n = 40) E. coli and urinary tract infection (n = 40) strains and normal flora E. coli strains (n = 40). Sorbitol negativity was common only in EHEC, particularly among strains from severe clinical infections. All 19 EHEC strains from patients with hemolytic-uremic syndrome, irrespective of O:H serotype or Shiga-like toxin genotype, were sorbitol negative.

  9. DIARRHEA, UROSEPSIS AND HEMOLYTIC UREMIC SYNDROME CAUSED BY THE SAME HETEROPATHOGENIC ESCHERICHIA COLI STRAIN

    NARCIS (Netherlands)

    Ang, C. Wim; Bouts, Antonia H. M.; Rossen, John W. A.; Van der Kuip, Martijn; Van Heerde, Marc; Bokenkamp, Arend

    2016-01-01

    We describe an 8-month-old girl with diarrhea, urosepsis and hemolytic uremic syndrome caused by Escherichia coli. Typing of cultured E. coli strains from urine and blood revealed the presence of virulence factors from multiple pathotypes of E. coli. This case exemplifies the genome plasticity of E.

  10. Proteomic differences between Escherichia coli strains that cause transient versus persistent intramammary infections [abstract

    Science.gov (United States)

    Escherichia coli is a leading cause of bacterial mastitis in dairy cattle. Typically this infection is transient in nature and lasts 2-3 days. However, in a minority of cases, E. coli can cause a persistent intramammary infection. The mechanisms that enable certain strains of E. coli to cause a p...

  11. Enhanced succinate production from glycerol by engineered Escherichia coli strains.

    Science.gov (United States)

    Li, Qing; Wu, Hui; Li, Zhimin; Ye, Qin

    2016-10-01

    In this study, an engineered strain Escherichia coli MLB (ldhA(-)pflB(-)) was constructed for production of succinate from glycerol. The succinate yield was 0.37mol/mol in anaerobic culture, however, the growth and glycerol consumption rates were very slow, resulting in a low succinate level. Two-stage fermentation was performed in flasks, and the succinate yield reached 0.93mol/mol, but the succinate titer was still low. Hence, overexpression of malate dehydrogenase, malic enzyme, phosphoenolpyruvate (PEP) carboxylase and PEP carboxykinase (PCK) from E. coli, and pyruvate carboxylase from Corynebacterium glutamicum in MLB was investigated for improving succinate production. Overexpression of PCK resulted in remarkable enhancement of glycerol consumption and succinate production. In flask experiments, the succinate concentration reached 118.1mM, and in a 1.5-L bioreactor the succinate concentration further increased to 360.2mM. The highest succinate yield achieved 0.93mol/mol, which was 93% of the theoretical yield, in the anaerobic stage.

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

    Science.gov (United States)

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

    1998-03-01

    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.

  13. Draft Genome Sequences of Five Novel Polyketide Synthetase-Containing Mouse Escherichia coli Strains

    Science.gov (United States)

    Mannion, Anthony; Shen, Zeli; Feng, Yan; Garcia, Alexis

    2016-01-01

    We report herein the draft genomes of five novel Escherichia coli strains isolated from surveillance and experimental mice housed at MIT and the Whitehead Institute and describe their genomic characteristics in context with the polyketide synthetase (PKS)-containing pathogenic E. coli strains NC101, IHE3034, and A192PP.

  14. Typical enteroaggregative Escherichia coli is the most prevalent pathotype among E. coli strains causing diarrhea in Mongolian children.

    Science.gov (United States)

    Sarantuya, Jav; Nishi, Junichiro; Wakimoto, Naoko; Erdene, Shirchin; Nataro, James P; Sheikh, Jalaluddin; Iwashita, Mayumi; Manago, Kunihiro; Tokuda, Koichi; Yoshinaga, Masao; Miyata, Koichiro; Kawano, Yoshifumi

    2004-01-01

    Diarrhea remains one of the main sources of morbidity and mortality in the world, and a large proportion is caused by diarrheagenic Escherichia coli. In Mongolia, the epidemiology of diarrheagenic E. coli has not been well studied. A total of 238 E. coli strains from children with sporadic diarrhea and 278 E. coli strains from healthy children were examined by PCR for 10 virulence genes: enteropathogenic E. coli (EPEC) eae, tir, and bfpA; enterotoxigenic E. coli (ETEC) lt and st; enteroinvasive E. coli (EIEC) ipaH; enterohemorragic E. coli stx1 and stx2; and enteroaggregative E. coli (EAEC) aggR and astA. EAEC strains without AggR were identified by the HEp-2 cell adherence test. The detection of EAEC, ETEC, EPEC, and EIEC was significantly associated with diarrhea. The incidence of EAEC (15.1%), defined by either a molecular or a phenotypic assay, was higher in the diarrheal group than any other category (0 to 6.0%). The incidence of AggR-positive EAEC in the diarrheal group was significantly higher than in the control group (8.0 versus 1.4%; P = 0.0004), while that of AggR-negative EAEC was not (7.1 versus 4.3%). Nineteen AggR-positive EAEC strains harbored other EAEC virulence genes-aggA, 2 (5.5%); aafA, 4 (11.1%); agg-3a, 5 (13.8%); aap, 8 (22.2%); aatA, 11 (30.5%); capU, 9 (25.0%); pet, 6 (16.6%); and set, 3 (8.3%)-and showed 15 genotypes. EAEC may be an important pathogen of sporadic diarrhea in Mongolian children. Genetic analysis showed the heterogeneity of EAEC but illustrated the importance of the AggR regulon (denoting typical EAEC) as a marker for virulent EAEC strains.

  15. Virulence Potential of Escherichia coli Strains Causing Asymptomatic Bacteriuria during Pregnancy ▿

    Science.gov (United States)

    Lavigne, Jean-Philippe; Boutet-Dubois, Adeline; Laouini, Dorsaf; Combescure, Christophe; Bouziges, Nicole; Marès, Pierre; Sotto, Albert

    2011-01-01

    We compared the virulence properties of a collection of asymptomatic bacteriuria (ABU) Escherichia coli strains to urinary tract infection (UTI) strains isolated from pregnant women in a university hospital over 1 year. The in vitro and in vivo studies suggest that ABU strains presented a virulence behavior similar to that of strains isolated from cases of cystitis. PMID:21918033

  16. Virulence potential of Escherichia coli strains causing asymptomatic bacteriuria during pregnancy.

    Science.gov (United States)

    Lavigne, Jean-Philippe; Boutet-Dubois, Adeline; Laouini, Dorsaf; Combescure, Christophe; Bouziges, Nicole; Marès, Pierre; Sotto, Albert

    2011-11-01

    We compared the virulence properties of a collection of asymptomatic bacteriuria (ABU) Escherichia coli strains to urinary tract infection (UTI) strains isolated from pregnant women in a university hospital over 1 year. The in vitro and in vivo studies suggest that ABU strains presented a virulence behavior similar to that of strains isolated from cases of cystitis.

  17. Virulence Potential of Escherichia coli Strains Causing Asymptomatic Bacteriuria during Pregnancy ▿

    OpenAIRE

    Lavigne, Jean-Philippe; Boutet-Dubois, Adeline; Laouini, Dorsaf; Combescure, Christophe; Bouziges, Nicole; Marès, Pierre; Sotto, Albert

    2011-01-01

    We compared the virulence properties of a collection of asymptomatic bacteriuria (ABU) Escherichia coli strains to urinary tract infection (UTI) strains isolated from pregnant women in a university hospital over 1 year. The in vitro and in vivo studies suggest that ABU strains presented a virulence behavior similar to that of strains isolated from cases of cystitis.

  18. Metabolic Modeling of Common Escherichia coli Strains in Human Gut Microbiome

    Directory of Open Access Journals (Sweden)

    Yue-Dong Gao

    2014-01-01

    Full Text Available The recent high-throughput sequencing has enabled the composition of Escherichia coli strains in the human microbial community to be profiled en masse. However, there are two challenges to address: (1 exploring the genetic differences between E. coli strains in human gut and (2 dynamic responses of E. coli to diverse stress conditions. As a result, we investigated the E. coli strains in human gut microbiome using deep sequencing data and reconstructed genome-wide metabolic networks for the three most common E. coli strains, including E. coli HS, UTI89, and CFT073. The metabolic models show obvious strain-specific characteristics, both in network contents and in behaviors. We predicted optimal biomass production for three models on four different carbon sources (acetate, ethanol, glucose, and succinate and found that these stress-associated genes were involved in host-microbial interactions and increased in human obesity. Besides, it shows that the growth rates are similar among the models, but the flux distributions are different, even in E. coli core reactions. The correlations between human diabetes-associated metabolic reactions in the E. coli models were also predicted. The study provides a systems perspective on E. coli strains in human gut microbiome and will be helpful in integrating diverse data sources in the following study.

  19. Towards a metabolic engineering strain "commons": an Escherichia coli platform strain for ethanol production.

    Science.gov (United States)

    Woodruff, Lauren B A; May, Brian L; Warner, Joseph R; Gill, Ryan T

    2013-05-01

    In the genome-engineering era, it is increasingly important that researchers have access to a common set of platform strains that can serve as debugged production chassis and the basis for applying new metabolic engineering strategies for modeling and characterizing flux, engineering complex traits, and optimizing overall performance. Here, we describe such a platform strain of E. coli engineered for ethanol production. Starting with a fully characterized host strain (BW25113), we site-specifically integrated the genes required for homoethanol production under the control of a strong inducible promoter into the genome and deleted the genes encoding four enzymes from competing pathways. This strain is capable of producing >30 g/L of ethanol in minimal media with production or tolerance was lost when grown under production conditions. Thus, our findings reinforce the need for a metabolic engineering "commons" that could provide a set of platform strains for use in more sophisticated genome-engineering strategies. Towards this end, we have made this production strain available to the scientific community.

  20. Behavior of non-O157 Shiga toxin-producing Escherichia coli, enteroinvasive E. coli, enteropathogenic E. coli, and enterotoxigenic E. coli strains on alfalfa sprouts.

    Science.gov (United States)

    Gómez-Aldapa, Carlos A; Rangel-Vargas, Esmeralda; Torres-Vitela, M Del Refugio; Villarruel-López, Angélica; Castro-Rosas, Javier

    2013-08-01

    Data about the behavior of non-O157 Shiga toxin-producing Escherichia coli (non-O157 STEC), enteroinvasive E. coli (EIEC), enterotoxigenic E. coli (ETEC), and enteropathogenic E. coli (EPEC) on seeds and alfalfa sprouts are not available. The behavior of STEC, EIEC, ETEC, and EPEC was determined during germination and sprouting of alfalfa seeds at 20 ± 2°C and 30 ± 2°C and on alfalfa sprouts at 3 ± 2°C. When alfalfa seeds were inoculated with STEC, EIEC, ETEC, or EPEC strains, all these diarrheagenic E. coli pathotypes (DEPs) grew during germination and sprouting of seeds, reaching counts of approximately 5 and 6 log CFU/g after 1 day at 20 ± 2°C and 30 ± 2°C, respectively. However, when the sprouts were inoculated after 1 day of seed germination and stored at 20 ± 2°C or 30 ± 2°C, no growth was observed for any DEP during sprouting at 20 ± 2°C or 30 ± 2°C for 9 days. Refrigeration reduced significantly (P < 0.0.5) the number of viable DEPs on sprouts after 20 days in storage; nevertheless, these decreases have no practical significance for the safety of the sprouts.

  1. Antibiotic resistant Esherichia coli strains from seafood and its susceptibility to seaweed extracts

    Institute of Scientific and Technical Information of China (English)

    Subramanian Kumaran; Balaraman Deivasigamani; Kumarappan Alagappan; Mannikam Sakthivel; Rajamani Karthikeyan

    2010-01-01

    Objective: To determine the prevalence and antibiotic resistance of Escherichia coli (E.coli) , in seafood obtained from Cuddalore and Parangipettai fish landing centres. Also, to identify the susceptibility of E. coli against predominant seaweeds red alga Kappaphycusalvarezii (K. alvarezii) and brown alga Padina boergessenii (P. boergessenii) extracts as sulfated polysaccharides and polyphenols respectively. Methods: A total of 48 samples (Two stations Cuddalore and Parangipettai, Tamil Nadu, India). Sampling area are fish landing centre where fishes caught from sea and estuary, seafood processing plants (packing and ice packed fishes) and local fish markets (fish samples). After isolation totally 80 strains were analyzed for its antimicrobial resistance and sensitivity against commercially 10 antibiotics. The ampicillin resistant E. coli CE21 was identified through molecular techniques as 16S rDNA sequencing. Two seaweeds K. alvarezii and P. boergessenii were screened for antibacterial activity against 12 antibiotic resistant E. coli strains. Results: Totally 48 swabbed samples from two different fish handling area were characterized for total bacterial and E. coli count. Mostly, the E. coli strains were isolated from fish local market and seafood processing plants before and after packaging process. In that maximum 56.25% strains were resistant to ampicillin and the minimum 2.5% strains were resistant to chloramphenicol. Therefore, the E. coli CE21 was identified through molecular techniques E. coli (GenBank accession number GU065251), The MIC value for polyphenol extract was slightly less than sulfated polysaccharides. E. coli strain isolated from Parangipettai was considerably increased MIC value that Cuddalore. Conclusions:The polyphenol and sulfated polysaccharides showed promising inhibitory response against all antimicrobial resistant E. coli strains and in particular the inhibitory response of ampicillin resistant E. coli.

  2. Pathogenic Escherichia coli strain discrimination using laser-induced breakdown spectroscopy

    Science.gov (United States)

    Diedrich, Jonathan; Rehse, Steven J.; Palchaudhuri, Sunil

    2007-07-01

    A pathogenic strain of bacteria, Escherichia coli O157:H7 (enterohemorrhagic E. coli or EHEC), has been analyzed by laser-induced breakdown spectroscopy (LIBS) with nanosecond pulses and compared to three nonpathogenic E. coli strains: a laboratory strain of K-12 (AB), a derivative of the same strain termed HF4714, and an environmental strain, E. coli C (Nino C). A discriminant function analysis (DFA) was performed on the LIBS spectra obtained from live colonies of all four strains. Utilizing the emission intensity of 19 atomic and ionic transitions from trace inorganic elements, the DFA revealed significant differences between EHEC and the Nino C strain, suggesting the possibility of identifying and discriminating the pathogenic strain from commonly occurring environmental strains. EHEC strongly resembled the two K-12 strains, in particular, HF4714, making discrimination between these strains difficult. DFA was also used to analyze spectra from two of the nonpathogenic strains cultured in different media: on a trypticase soy (TS) agar plate and in a liquid TS broth. Strains cultured in different media were identified and effectively discriminated, being more similar than different strains cultured in identical media. All bacteria spectra were completely distinct from spectra obtained from the nutrient medium or ablation substrate alone. The ability to differentiate strains prepared and tested in different environments indicates that matrix effects and background contaminations do not necessarily preclude the use of LIBS to identify bacteria found in a variety of environments or grown under different conditions.

  3. Lytic and lysogenic infection of diverse Escherichia coli and Shigella strains with a verocytotoxigenic bacteriophage.

    Science.gov (United States)

    James, C E; Stanley, K N; Allison, H E; Flint, H J; Stewart, C S; Sharp, R J; Saunders, J R; McCarthy, A J

    2001-09-01

    A verocytotoxigenic bacteriophage isolated from a strain of enterohemorrhagic Escherichia coli O157, into which a kanamycin resistance gene (aph3) had been inserted to inactivate the verocytotoxin gene (vt2), was used to infect Enterobacteriaceae strains. A number of Shigella and E. coli strains were susceptible to lysogenic infection, and a smooth E. coli isolate (O107) was also susceptible to lytic infection. The lysogenized strains included different smooth E. coli serotypes of both human and animal origin, indicating that this bacteriophage has a substantial capacity to disseminate verocytotoxin genes. A novel indirect plaque assay utilizing an E. coli recA441 mutant in which phage-infected cells can enter only the lytic cycle, enabling detection of all infective phage, was developed.

  4. Biofilm-Exclusion of Uropathogenic Bacteria by Selected Asymptomatic Bacteriuria Escherichia Coli Strains

    DEFF Research Database (Denmark)

    Ferriéres, L.; Hancock, Viktoria; Klemm, Per

    2007-01-01

    . In contrast to uropathogenic E coli (UPEC), which cause symptomatic urinary tract infection, asymptomatic bacteriuria (ABU) strains are associated with essentially symptom-free infections. Here the biofilm-forming capacity on abiotic surfaces of selected E coli ABU strains and UPEC strains in human urine...... was investigated. It was found that there is a strong bias for biofilm formation by the ABU strains. Not only were the ABU strains significantly better biofilm formers than UPEC strains, they were also able to out-compete UPEC strains as well as uropathogenic strains of Klebsiella spp. during biofilm formation....... The results support the notion of bacterial prophylaxis employing selected ABU strains to eliminate UPEC strains and other pathogens in patients prone to recalcitrant infections....

  5. Shigella strains are not clones of Escherichia coli but sister species in the genus Escherichia.

    Science.gov (United States)

    Zuo, Guanghong; Xu, Zhao; Hao, Bailin

    2013-02-01

    Shigella species and Escherichia coli are closely related organisms. Early phenotyping experiments and several recent molecular studies put Shigella within the species E. coli. However, the whole-genome-based, alignment-free and parameter-free CVTree approach shows convincingly that four established Shigella species, Shigella boydii, Shigella sonnei, Shigella felxneri and Shigella dysenteriae, are distinct from E. coli strains, and form sister species to E. coli within the genus Escherichia. In view of the overall success and high resolution power of the CVTree approach, this result should be taken seriously. We hope that the present report may promote further in-depth study of the Shigella-E. coli relationship.

  6. Biofilm formation by asymptomatic and virulent urinary tract infectious Escherichia coli strains

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Ferrieres, Lionel; Klemm, Per

    2007-01-01

    Escherichia coli is the most common organism associated with asymptomatic bacteriuria (ABU) in humans. In contrast to uropathogenic E. coli (UPEC) that cause symptomatic urinary tract infection, very little is known about the mechanisms by which these strains colonize the urinary tract. Here, we...... have investigated the biofilm-forming capacity on abiotic surfaces of groups of ABU strains and UPEC strains in human urine. We found that there is a strong bias; ABU strains were significantly better biofilm formers than UPEC strains. Our data suggest that biofilm formation in urinary tract infectious...

  7. Diffusely Adhering Escherichia coli Strains Induce Attaching and Effacing Phenotypes and Secrete Homologs of Esp Proteins

    OpenAIRE

    Beinke, Christina; Laarmann, Sven; Wachter, Clemens; Karch, Helge; Greune, Lilo; Schmidt, M. Alexander

    1998-01-01

    Recent epidemiological studies indicate that Escherichia coli strains which exhibit the diffuse-adherence phenotype (DAEC strains) represent a potential cause of diarrhea in infants. We investigated the interaction of DAEC strains isolated from diarrhea patients in Brazil and in Germany with epithelial cells in tissue culture. The investigated strains were identified as DAEC strains by (i) their attachment pattern, (ii) presence of genes associated with the Dr family of adhesins, and (iii) la...

  8. Genome Sequence of Avian Escherichia coli Strain IHIT25637, an Extraintestinal Pathogenic E. coli Strain of ST131 Encoding Colistin Resistance Determinant MCR-1.

    Science.gov (United States)

    Ewers, Christa; Göttig, Stephan; Bülte, Maria; Fiedler, Sophie; Tietgen, Manuela; Leidner, Ursula; Heydel, Carsten; Bauerfeind, Rolf; Semmler, Torsten

    2016-09-01

    Sequence type 131 (ST131) is one of the predominant Escherichia coli lineages among extraintestinal pathogenic E. coli (ExPEC) that causes a variety of diseases in humans and animals and frequently shows multidrug resistance. Here, we report the first genome sequence of an ST131-ExPEC strain from poultry carrying the plasmid-encoded colistin resistance gene mcr-1. Copyright © 2016 Ewers et al.

  9. Genotypic and phenotypic characterisation of Escherichia coli strains associated with porcine pyelonephritis

    DEFF Research Database (Denmark)

    Krag, L.; Hancock, Viktoria; Aalbæk, B.

    2009-01-01

    type when it comes to human isolates. However, relatively little has been done to characterise the corresponding porcine strains. On this background we have analysed 20 porcine pyelonephritis E coli strains isolated from infected pig kidneys. The strains were quite distinct from that of human...

  10. [Current antibiotic resistance profile of uropathogenic Escherichia coli strains and therapeutic consequences].

    Science.gov (United States)

    El Bouamri, M C; Arsalane, L; Kamouni, Y; Yahyaoui, H; Bennouar, N; Berraha, M; Zouhair, S

    2014-12-01

    Urinary tract infections (UTI) are a very common reason for consultation and prescription in current practice. Excessive or inappropriate use of antibiotics in treating urinary tract infections is responsible for the emergence and spread of multiresistant uropathogenic bacteria. To evaluate the isolation frequency and antibiotic resistance of uropathogenic Escherichia coli strains isolated at the Marrakech region. We conducted a retrospective study over a period of three years (from 1st January 2010 to 31 December 2012). It included all non-redundant uropathogenic E. coli strains isolated in the microbiology laboratory of the Avicenne hospital of Marrakech, Morocco. During this study, 1472 uropathogenic enterobacteriaceae were isolated including 924 non-repetitive E. coli strains, an overall isolation frequency of 63%. Antibiotic resistance of isolated E. coli strains showed resistance rates to amoxicillin (65%), sulfamethoxazole-triméthropime (55%), amoxicillin-clavulanic acid (43%), ciprofloxacin (22%), gentamicin (14%), nitrofurans (11%), amikacin (8%) and fosfomycin (7%). The number of E. coli strains resistant to C3G by ESBL production was 67, an average frequency of 4.5% of all isolated uropathogenic enterobacteria. The associated antibiotic resistance in the case of ESBL-producing E. coli were 82% for ciprofloxacin, 76% for sulfamethozole trimethoprim, 66% for gentamicin and 56% for amikacin. No resistance to imipenem was recorded for the isolated E. coli strains, which represents an imipenem sensitivity of 100%. Antibiotic resistance of uropathogenic E. coli strains limits treatment options and therefore constitutes a real public health problem. The regular updating of antibiotic susceptibility statistics of E. coli strains allows a better adaptation of the probabilistic antibiotic therapy to local epidemiological data. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

  11. Comparative Genomics and Characterization of Hybrid Shigatoxigenic and Enterotoxigenic Escherichia coli (STEC/ETEC) Strains

    Science.gov (United States)

    Nyholm, Outi; Halkilahti, Jani; Wiklund, Gudrun; Okeke, Uche; Paulin, Lars; Auvinen, Petri; Haukka, Kaisa; Siitonen, Anja

    2015-01-01

    Background Shigatoxigenic Escherichia coli (STEC) and enterotoxigenic E. coli (ETEC) cause serious foodborne infections in humans. These two pathogroups are defined based on the pathogroup-associated virulence genes: stx encoding Shiga toxin (Stx) for STEC and elt encoding heat-labile and/or est encoding heat-stable enterotoxin (ST) for ETEC. The study investigated the genomics of STEC/ETEC hybrid strains to determine their phylogenetic position among E. coli and to define the virulence genes they harbor. Methods The whole genomes of three STEC/ETEC strains possessing both stx and est genes were sequenced using PacBio RS sequencer. Two of the strains were isolated from the patients, one with hemolytic uremic syndrome, and one with diarrhea. The third strain was of bovine origin. Core genome analysis of the shared chromosomal genes and comparison with E. coli and Shigella spp. reference genomes was performed to determine the phylogenetic position of the STEC/ETEC strains. In addition, a set of virulence genes and ETEC colonization factors were extracted from the genomes. The production of Stx and ST were studied. Results The human STEC/ETEC strains clustered with strains representing ETEC, STEC, enteroaggregative E. coli, and commensal and laboratory-adapted E. coli. However, the bovine STEC/ETEC strain formed a remote cluster with two STECs of bovine origin. All three STEC/ETEC strains harbored several other virulence genes, apart from stx and est, and lacked ETEC colonization factors. Two STEC/ETEC strains produced both toxins and one strain Stx only. Conclusions This study shows that pathogroup-associated virulence genes of different E. coli can co-exist in strains originating from different phylogenetic lineages. The possibility of virulence genes to be associated with several E. coli pathogroups should be taken into account in strain typing and in epidemiological surveillance. Development of novel hybrid E. coli strains may cause a new public health risk, which

  12. Comparative Genomics and Characterization of Hybrid Shigatoxigenic and Enterotoxigenic Escherichia coli (STEC/ETEC Strains.

    Directory of Open Access Journals (Sweden)

    Outi Nyholm

    Full Text Available Shigatoxigenic Escherichia coli (STEC and enterotoxigenic E. coli (ETEC cause serious foodborne infections in humans. These two pathogroups are defined based on the pathogroup-associated virulence genes: stx encoding Shiga toxin (Stx for STEC and elt encoding heat-labile and/or est encoding heat-stable enterotoxin (ST for ETEC. The study investigated the genomics of STEC/ETEC hybrid strains to determine their phylogenetic position among E. coli and to define the virulence genes they harbor.The whole genomes of three STEC/ETEC strains possessing both stx and est genes were sequenced using PacBio RS sequencer. Two of the strains were isolated from the patients, one with hemolytic uremic syndrome, and one with diarrhea. The third strain was of bovine origin. Core genome analysis of the shared chromosomal genes and comparison with E. coli and Shigella spp. reference genomes was performed to determine the phylogenetic position of the STEC/ETEC strains. In addition, a set of virulence genes and ETEC colonization factors were extracted from the genomes. The production of Stx and ST were studied.The human STEC/ETEC strains clustered with strains representing ETEC, STEC, enteroaggregative E. coli, and commensal and laboratory-adapted E. coli. However, the bovine STEC/ETEC strain formed a remote cluster with two STECs of bovine origin. All three STEC/ETEC strains harbored several other virulence genes, apart from stx and est, and lacked ETEC colonization factors. Two STEC/ETEC strains produced both toxins and one strain Stx only.This study shows that pathogroup-associated virulence genes of different E. coli can co-exist in strains originating from different phylogenetic lineages. The possibility of virulence genes to be associated with several E. coli pathogroups should be taken into account in strain typing and in epidemiological surveillance. Development of novel hybrid E. coli strains may cause a new public health risk, which challenges the

  13. EMERGENCE OF MULTI DRUG RESISTANT STRAINS OF E. COLI ISOLATED FROM URINARY TRACT INFECTION IN NAMAKKAL

    OpenAIRE

    2012-01-01

    In this study we evaluated the multi drug resistant strains of Escherichia coli isolated from urinary tract infection in different age groups in Namakkal. Totally 153 isolates of E.coli were obtained from 700 urine samples. Present study indicates that there is a high prevalence of multidrug resistant E. coli in female 24.34% followed by male 18.14%. After confirmation, isolates were performed to resistance patterns of different antibiotics were determined by standard disk diffusion method. T...

  14. Occurrence and characterization of verocytotoxigenic Escherichia coli (VTEC) strains from dairy farms in Trinidad.

    Science.gov (United States)

    Roopnarine, R R; Ammons, D; Rampersad, J; Adesiyun, A A

    2007-01-01

    A cross-sectional study was conducted to determine the prevalence and characteristics of verocytotoxigenic Escherichia coli (VTEC) on 25 dairy farms each located in Waller field and Carlsen field farming areas in Trinidad. On each selected farm, faecal samples were collected from milking cows, calves and humans; rectal swabs were obtained from pet farm dogs; bulk milk was sampled as well as effluent from the milking parlour. Escherichia coli was isolated from all sources on selective media using standard methods. Isolates of E. coli were subjected to slide agglutination test using E. coli O157 antiserum, vero cell cytotoxicity assay to detect verocytotoxin (VT) and heat labile toxin (LT) production, the polymerase chain reaction (PCR) to detect VT genes, and the dry spot test to screen for E. coli O157 and non-O157 strains. In addition, faecal samples from animal and human sources were tested for VT genes using PCR. Of a total of 933 E. coli isolates tested by the slide test, eight (0.9%) were positive for the O157 strain. The vero cell cytotoxicity assay detected VT-producing strains of E. coli in 16.6%, 14.6%, 3.2% and 7.1% of isolates from cows, calves, farm dogs and humans respectively (P detected amongst isolates of E. coli from calves (10.8%) and the lowest (0.0%) amongst isolates from humans and bulk milk (P detected in 62.3%, 4.9% and 1.6% respectively (P Detection of VTEC strains in milk and dairy animals poses a health risk to consumers of milk originating from these farms. In addition, the demonstration of VTEC strains in humans, VT gene in faecal samples and E. coli isolates as well as non-O157 VTEC strains of E. coli are being documented for the first time in the country.

  15. Comparison of Escherichia coli Strains Recovered from Human Cystitis and Pyelonephritis Infections in Transurethrally Challenged Mice

    Science.gov (United States)

    Johnson, David E.; Lockatell, C. Virginia; Russell, Robert G.; Hebel, J. Richard; Island, Michael D.; Stapleton, Ann; Stamm, Walter E.; Warren, John W.

    1998-01-01

    Urinary tract infection, most frequently caused by Escherichia coli, is one of the most common bacterial infections in humans. A vast amount of literature regarding the mechanisms through which E. coli induces pyelonephritis has accumulated. Although cystitis accounts for 95% of visits to physicians for symptoms of urinary tract infections, few in vivo studies have investigated possible differences between E. coli recovered from patients with clinical symptoms of cystitis and that from patients with symptoms of pyelonephritis. Epidemiological studies indicate that cystitis-associated strains appear to differ from pyelonephritis-associated strains in elaboration of some putative virulence factors. With transurethrally challenged mice we studied possible differences using three each of the most virulent pyelonephritis and cystitis E. coli strains in our collection. The results indicate that cystitis strains colonize the bladder more rapidly than do pyelonephritis strains, while the rates of kidney colonization are similar. Cystitis strains colonize the bladder in higher numbers, induce more pronounced histologic changes in the bladder, and are more rapidly eliminated from the mouse urinary tract than pyelonephritis strains. These results provide evidence that cystitis strains differ from pyelonephritis strains in this model, that this model is useful for the study of the uropathogenicity of cystitis strains, and that it would be unwise to use pyelonephritis strains to study putative virulence factors important in the development of cystitis. PMID:9632566

  16. Isolation of Escherichia coli 0157:H7 strain from fecal samples of zoo animal.

    Science.gov (United States)

    Mohammed Hamzah, Aseel; Mohammed Hussein, Aseel; Mahmoud Khalef, Jenan

    2013-01-01

    The isolation and characterization of Escherichia coli O157:H7 strains from 22 out of 174 fecal samples from petting zoo animals representing twenty-two different species (camel, lion, goats, zebra, bear, baboon monkey, Siberian monkey, deer, elk, llama, pony, horses, fox, kangaroo, wolf, porcupine, chickens, tiger, ostrich, hyena, dogs, and wildcats) were investigated. One petting Al-Zawraa zoological society of Baghdad was investigated for E. coli O157:H7 over a 16-month period that spanned two summer and two autumn seasons. Variation in the occurrence of E. coli O157:H7-positive petting zoo animals was observed, with animals being culture positive only in the summer months but not in the spring, autumn, or winter. E. coli O157:H7 isolates were distinguished by agglutination with E. coli O157:H7 latex reagent (Oxoid), identified among the isolates, which showed that multiple E. coli strains were isolated from one petting zoo animal, in which a single animal simultaneously shed multiple E. coli strains; E. coli O157:H7 was isolated only by selective enrichment culture of 2 g of petting zoo animal feces. In contrast, strains other than O157:H7 were cultured from feces of petting zoo animals without enrichment.

  17. Pathogenic potential of Escherichia coli clinical strains from orthopedic implant infections towards human osteoblastic cells

    Science.gov (United States)

    Crémet, Lise; Broquet, Alexis; Brulin, Bénédicte; Jacqueline, Cédric; Dauvergne, Sandie; Brion, Régis; Asehnoune, Karim; Corvec, Stéphane; Heymann, Dominique; Caroff, Nathalie

    2015-01-01

    Escherichia coli is one of the first causes of Gram-negative orthopedic implant infections (OII), but little is known about the pathogenicity of this species in such infections that are increasing due to the ageing of the population. We report how this pathogen interacts with human osteoblastic MG-63 cells in vitro, by comparing 20 OII E. coli strains to two Staphylococcus aureus and two Pseudomonas aeruginosa strains. LDH release assay revealed that 6/20 (30%) OII E. coli induced MG-63 cell lysis whereas none of the four control strains was cytotoxic after 4 h of coculture. This high cytotoxicity was associated with hemolytic properties and linked to hlyA gene expression. We further showed by gentamicin protection assay and confocal microscopy that the non-cytotoxic E. coli were not able to invade MG-63 cells unlike S. aureus strains (internalization rate coli versus 8.88 ± 2.31% and 4.60 ± 0.42% for both S. aureus). The non-cytotoxic E. coli also demonstrated low adherence rates (coli eliciting higher IL-6 and TNF-α mRNA expression in the osteoblastic cells. Either highly cytotoxic or slightly invasive OII E. coli do not show the same infection strategies as S. aureus towards osteoblasts. PMID:26333570

  18. Genome Sequences of Two Bovine Mastitis-Causing Escherichia coli Strains

    OpenAIRE

    Kempf, Florent; Loux, Valentin; Germon, Pierre

    2015-01-01

    Escherichia coli is one of the main pathogenic agents causing inflammatory infections in the bovine udder. Here, we report the draft genome sequences of two strains isolated from different cases of clinical mastitis.

  19. Origins of the E. coli strain causing an outbreak of hemolytic-uremic syndrome in Germany

    DEFF Research Database (Denmark)

    Rasko, David A; Webster, Dale R; Sahl, Jason W

    2011-01-01

    A large outbreak of diarrhea and the hemolytic-uremic syndrome caused by an unusual serotype of Shiga-toxin-producing Escherichia coli (O104:H4) began in Germany in May 2011. As of July 22, a large number of cases of diarrhea caused by Shiga-toxin-producing E. coli have been reported--3167 without...... the hemolytic-uremic syndrome (16 deaths) and 908 with the hemolytic-uremic syndrome (34 deaths)--indicating that this strain is notably more virulent than most of the Shiga-toxin-producing E. coli strains. Preliminary genetic characterization of the outbreak strain suggested that, unlike most of these strains......, it should be classified within the enteroaggregative pathotype of E. coli....

  20. Molecular typing of uropathogenic E. coli strains by the ERIC-PCR method.

    Science.gov (United States)

    Ardakani, Maryam Afkhami; Ranjbar, Reza

    2016-04-01

    Escherichia coli (E. coli) is the most common cause of urinary infections in hospitals. The aim of this study was to evaluate the ERIC-PCR method for molecular typing of uropathogenic E. coli strains isolated from hospitalized patients. In a cross sectional study, 98 E. coli samples were collected from urine samples taken from patients admitted to Baqiyatallah Hospital from June 2014 to January 2015. The disk agar diffusion method was used to determine antibiotic sensitivity. DNA proliferation based on repetitive intergenic consensus was used to classify the E. coli strains. The products of proliferation were electrophoresed on 1.5% agarose gel, and their dendrograms were drawn. The data were analyzed by online Insillico software. The method used in this research proliferated numerous bands (4-17 bands), ranging from 100 to 3000 base pairs. The detected strains were classified into six clusters (E1-E6) with 70% similarity between them. In this study, uropathogenic E. coli strains belonged to different genotypic clusters. It was found that ERIC-PCR had good differentiation power for molecular typing of uropathogenic E. coli strains isolated from the patients in the study.

  1. Genome Sequences of Two Copper-Resistant Escherichia coli Strains Isolated from Copper-Fed Pigs

    DEFF Research Database (Denmark)

    Lüthje, Freja L.; Hasman, Henrik; Aarestrup, Frank Møller;

    2014-01-01

    The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances.......The draft genome sequences of two copper-resistant Escherichia coli strains were determined. These had been isolated from copper-fed pigs and contained additional putative operons conferring copper and other metal and metalloid resistances....

  2. Draft Genome Sequence of Brazilian Escherichia coli Uropathogenic Strain E2

    Science.gov (United States)

    Pieta, Luiza; Prichula, Janira; Sambrano, Gustavo E.; Soares, Renata; Caierão, Juliana; Frazzon, Jeverson; d’Azevedo, Pedro A.

    2016-01-01

    Escherichia coli is a common pathogen recovered from cystitis infections. In this report, we announce the draft genome sequence of strain E2 isolated from the urine specimen from a female patient in South Brazil. The genome assembly has 5,081,209 bp, a G+C content of 50.57%, and virulence factors associated with both enteroaggregative and uropathogenic E. coli strains.

  3. [Joint cultivation of Bacillus subtilis and Escherichia coli strains promising for obtaining complex probiotic].

    Science.gov (United States)

    Tsaruk'ianova, I G; Osadchaia, A I

    2007-01-01

    The ability of joint cultivation of Bacillus subtilis UCM B-5007 and Escherichia coli M-17 in subsurface conditions has been studied. These strains are available for creation of a new complex probiotic. Symbiotic relationships between these microorganisms were proved. Bacillus subtilis and Escherichia coli strains use different growth "strategy". The most optimum ratio of cultures (1:1) for growth, biomass accumulation, and for antagonism to test-cultures has been chosen.

  4. Plasmid profiling and antibiotics resisitance of Escherichia coli strains isolated from Mytilus galloprovincialis and seawater

    Directory of Open Access Journals (Sweden)

    Cumhur Avşar

    2014-09-01

    Full Text Available Objective: To investigate plasmid DNA profiles and the antibiotic resistance of a total of 41 strains of Escherichia coli (E. coli isolated from seawater and mussel collected from 15 different sampling stations in Sinop, Turkey. Methods: Most probable number technique was used for detection of E. coli. Antibiotic susceptibilities of the isolates were determined by the disc diffusion method. Plasmid DNA of the strains was extracted by the alkaline lyses procedure. Results: According to morphological and physiological properties, it was determined that the isolates belonged to E. coli species. Antibiotic susceptibility of the strains was determined against seven standard drugs using disc diffusion method. All isolates were resistant to bacitracin (100%, novobiocin (100%, ampicillin (12.5%, tetracycline (7.5%, ceftazidime (5% and imipenem (2.5%, respectively, whereas the strains were susceptible to polymyxin B (100%. The multiple antibiotic resistance values for the strains were found in range from 0.28 to 0.57. In addition, plasmid DNA analyses results confirmed that 22 strains harbored a single or more than two plasmids sized approximately between 24.500 to 1.618 bp. The high-size plasmid (14.700 bp was observed as common in 21 of all strains. Conclusions: As a result, our study indicated that the presence of antibiotic resistant E. coli strains in seawater and mussel might be potential risk for public health issue.

  5. Plasmid profiling and antibiotics resisitance of Escherichia coli strains isolated from Mytilus galloprovincialis and seawater

    Institute of Scientific and Technical Information of China (English)

    Cumhur Avşar; İsmet Berber

    2014-01-01

    Objective: To investigate plasmid DNA profiles and the antibiotic resistance of a total of 41 strains of Escherichia coli (E. coli) isolated from seawater and mussel collected from 15 different sampling stations in Sinop, Turkey. Methods: Most probable number technique was used for detection of E. coli. Antibiotic susceptibilities of the isolates were determined by the disc diffusion method. Plasmid DNA of the strains was extracted by the alkaline lyses procedure.Results:According to morphological and physiological properties, it was determined that the isolates belonged to E. coli species. Antibiotic susceptibility of the strains was determined against seven standard drugs using disc diffusion method. All isolates were resistant to bacitracin (100%), novobiocin (100%), ampicillin (12.5%), tetracycline (7.5%), ceftazidime (5%) and imipenem (2.5%), respectively, whereas the strains were susceptible to polymyxin B (100%). The multiple antibiotic resistance values for the strains were found in range from 0.28 to 0.57. In addition, plasmid DNA analyses results confirmed that 22 strains harbored a single or more than two plasmids sized approximately between 24.500 to 1.618 bp. The high-size plasmid (14.700 bp) was observed as common in 21 of all strains.Conclusions:As a result, our study indicated that the presence of antibiotic resistant E. coli strains in seawater and mussel might be potential risk for public health issue.

  6. Comparison of whole genome sequences from human and non-human Escherichia coli O26 strains

    Directory of Open Access Journals (Sweden)

    Keri N Norman

    2015-03-01

    Full Text Available Shiga toxin-producing Escherichia coli (STEC O26 is the second leading E. coli serogroup responsible for human illness outbreaks behind E. coli O157:H7. Recent outbreaks have been linked to emerging pathogenic O26:H11 strains harboring stx2 only. Cattle have been recognized as an important reservoir of O26 strains harboring stx1; however the reservoir of these emerging stx2 strains is unknown. The objective of this study was to identify nucleotide polymorphisms in human and cattle-derived strains in order to compare differences in polymorphism derived genotypes and virulence gene profiles between the two host species. Whole genome sequencing was performed on 182 epidemiologically unrelated O26 strains, including 109 human-derived strains and 73 non-human-derived strains. A panel of 289 O26 strains (241 STEC and 48 non-STEC was subsequently genotyped using a set of 283 polymorphisms identified by whole genome sequencing, resulting in 64 unique genotypes. Phylogenetic analyses identified seven clusters within the O26 strains. The seven clusters did not distinguish between isolates originating from humans or cattle; however, clusters did correspond with particular virulence gene profiles. Human and non-human-derived strains harboring stx1 clustered separately from strains harboring stx2, strains harboring eae, and non-STEC strains. Strains harboring stx2 were more closely related to non-STEC strains and strains harboring eae than to strains harboring stx1. The finding of human and cattle-derived strains with the same polymorphism derived genotypes and similar virulence gene profiles, provides evidence that similar strains are found in cattle and humans and transmission between the two species may occur.

  7. The construction of Streptomyces cyaneus genomic libraries in Escherichia coli is dependent upon the use of Mcr-deficient strains.

    Science.gov (United States)

    Wang, P; Harvey, S S; Sims, P F; Broda, P

    1992-09-21

    Streptomyces cyaneus genomic DNA ligated into either lambda phage or plasmid vectors was very inefficiently cloned into standard Escherichia coli host strains. However, the same material could be efficiently cloned using Mcr-deficient E. coli strains. These results suggest that the S. cyaneus genome contains 5-methylcytosine residues, some of which occur within the recognition sequences of the E. coli Mcr restriction system.

  8. Antibiotic Resistance Patterns in Enteric and Uropathogenic Strains of Escherichia Coli in Children

    Directory of Open Access Journals (Sweden)

    This paper should be cited as: Sedighi I, Alikhani MY, Nakhaee S, Karami P . [ Antibiotic Resistance Patterns in Enteric and Uropathogenic Strains of Escherichia Coli in Children ]. mlj goums . 201 4 ; 8 ( Suppl 4 : 42 - 48 [Article in Per sian] Sedi ghi, I. (MD

    2014-11-01

    Full Text Available Background and Objective: Escherichia coli is the most common cause of urinary tract infections in children and the leading cause of intra-abdominal infections (peritonitis and abscess followed intestinal injuries. Urinary tract infection, including cystitis and pyelonephritis, is a common childhood infection. E. coli causes more than 90 percent of the community acquired and 50% of hospital acquired urinary tract infections; therefore, the determination of E. coli antibiotic susceptibility is a paramount importance to clinical and epidemiological purposes. Material and Methods: In this cross-sectional study, 50 E. coli strains isolated from urine samples of children less than 7 years of age with urinary tract infections. They were compared for drug susceptibility testing by disc diffusion method with 50 strains of Escherichia coli isolated from stool samples of healthy children with the same age and sex pattern. Results: The actual amount of drug sensitivity of uropathogenic and intestinal Escherichia coli strains to amikacin was 94 and 100%, nitrofurantoin 90 and 88%, gentamicin 66 and 94%, cefixime 56 and 60%, nalidixic acid 38 and 44% and to cotrimoxazole 28 and 32%, respectively. Conclusion: the rate of resistance to gentamicin, Cefixime and nalidixic acid in urinary tract infection isolates were more than intestinal strains. The highest rate of drug resistance in urinary Escherichia coli isolates was associated with cotrimoxazole and the lowest one with amikacin.

  9. Optimized E. coli expression strain LOBSTR eliminates common contaminants from His-tag purification.

    Science.gov (United States)

    Andersen, Kasper R; Leksa, Nina C; Schwartz, Thomas U

    2013-11-01

    His-tag affinity purification is one of the most commonly used methods to purify recombinant proteins expressed in E. coli. One drawback of using the His-tag is the co-purification of contaminating histidine-rich E. coli proteins. We engineered a new E. coli expression strain, LOBSTR (low background strain), which eliminates the most abundant contaminants. LOBSTR is derived from the E. coli BL21(DE3) strain and carries genomically modified copies of arnA and slyD, whose protein products exhibit reduced affinities to Ni and Co resins, resulting in a much higher purity of the target protein. The use of LOBSTR enables the pursuit of challenging low-expressing protein targets by reducing background contamination with no additional purification steps, materials, or costs, and thus pushes the limits of standard His-tag purifications.

  10. CHARACTERIZATION OF EXTENDED-SPECTRUM Β-LACTAMASE-PRODUCING ESCHERICHIA COLI STRAINS ISOLATED FROM DAIRY PRODUCTS

    Directory of Open Access Journals (Sweden)

    Rahem Khoshbakht

    2014-02-01

    Full Text Available Extended-spectrum β-lactamases (ESBLs are enzymes that hydrolyze the β-lactam ring, and ESBL-producing E. coli has rapidly spread worldwide with pose a serious hazard for humans. The aim of this study was to determine the prevalence of ESBL producing E. coli and molecular evaluation of four ESBL-associated genes among E. coli strains isolated from milk and cheese in southern Iran. Antibiotic susceptibility test was carried out for a total of 150 isolates of E. coli, previously collected from dairy products. ESBL production was screened using a double-disc synergy test (DDST and presence of four ESBL genes (PER, VEB, TEM and CTX-M was tested using PCR. Among 150 E. coli strains 57 (38% isolates were identified as ESBL-producing strains. All ESBL positive isolates could be typed for one or more genes and the most prevalent ESBL-associated gene was CTX-M (80.7%. The PER gene was not present among isolates. Isolates showed high susceptibility to imipe¬nem and cefoxitin. The results showed the high prevalence of ESBL producing E. coli strains among dairy products and high occurrence of CTX-M-associated ESBL activity among isolates indicating the hazards of increasing the strains with antibiotic resistance which can transfer to human trough the dairy food products.

  11. Survival and resuscitation of ten strains of Campylobacter jejuni and Campylobacter coli under acid conditions

    NARCIS (Netherlands)

    Chaveerach, P.; Huurne, ter A.A.H.M.; Lipman, L.J.A.; Knapen, van F.

    2003-01-01

    The culturability of 10 strains of Campylobacter jejuni and Campylobacter coli was studied after the bacteria were exposed to acid conditions for various periods of time. Campylobacter cells could not survive 2 h under acid conditions (formic acid at pH 4). The 10 Campylobacter strains could not be

  12. Survival and resuscitation of ten strains of Campylobacter jejuni and Campylobacter coli under acid conditions

    NARCIS (Netherlands)

    Chaveerach, P.; Huurne, ter A.A.H.M.; Lipman, L.J.A.; Knapen, van F.

    2003-01-01

    The culturability of 10 strains of Campylobacter jejuni and Campylobacter coli was studied after the bacteria were exposed to acid conditions for various periods of time. Campylobacter cells could not survive 2 h under acid conditions (formic acid at pH 4). The 10 Campylobacter strains could not be

  13. Isolation and characterization of the E. coli membrane protein production strain Mutant56(DE3)

    NARCIS (Netherlands)

    Baumgarten, Thomas; Schlegel, Susan; Wagner, Samuel; Löw, Mirjam; Eriksson, Jonas; Bonde, Ida; Herrgård, Markus J; Heipieper, Hermann J; Nørholm, Morten H H; Slotboom, Dirk Jan; de Gier, Jan-Willem

    2017-01-01

    Membrane protein production is usually toxic to E. coli. However, using genetic screens strains can be isolated in which the toxicity of membrane protein production is reduced, thereby improving production yields. Best known examples are the C41(DE3) and C43(DE3) strains, which are both derived from

  14. Molecular screening of pathogenic Escherichia coli strains isolated from dairy neonatal calves in Cordoba province, Argentina.

    Science.gov (United States)

    Picco, Natalia Y; Alustiza, Fabrisio E; Bellingeri, Romina V; Grosso, María C; Motta, Carlos E; Larriestra, Alejandro J; Vissio, Claudina; Tiranti, Karina I; Terzolo, Horacio R; Moreira, Ana R; Vivas, Adriana B

    2015-01-01

    The aim of this study was to perform a current molecular characterization of bovine pathogenic Escherichia coli strains isolated from random samplings in Argentinean dairy farms. Rectal swabs were obtained from 395 (63.7%) healthy and 225 (36.3%) diarrheic calves, belonging to 45 dairy farms in Cordoba Province, Argentina. E. coli isolates were examined for virulence genes (f5, f41, f17, sta, stb, lt, eae, vt) using PCR and the prevalence of E. coli virulence profiles was spatially described in terms of spatial distribution. A total of 30.1% isolates were found to be positive for at least one of the virulence genes. Depending on the different gene combinations present, 11 virulence profiles were found. Most of the isolates analyzed had a single gene, and no combination of fimbrial and enterotoxin gene was predominant. There was no association between the frequency and distribution of E. coli virulence genes and calf health status. Most of the virulence profiles were compatible with ETEC strains and showed a homogeneous distribution over the sampled area. A clustering pattern for E. coli virulence profiles could not be recognized. This work provides updated information on the molecular characterization of pathogenic E. coli strains from dairy herds in Cordoba, Argentina. These findings would be important to formulate prevention programs and effective therapies for diarrhea in calves caused by E. coli.

  15. Genetic Evaluation of E. coli Strains Isolated from Asymptomatic Children with Neurogenic Bladders

    Directory of Open Access Journals (Sweden)

    John Kryger

    2015-01-01

    Full Text Available This study was conducted to describe the genetic profiles of E. coli that colonize asymptomatic pediatric neurogenic bladders. E. coli was isolated from 25 of 80 urine samples. Patients were excluded if they presented with symptomatic urinary tract infection or received treatment with antibiotics in the preceding three months. Multiplex PCR was performed to determine E. coli phylotype (A, B1, B2, and D and the presence of seven pathogenicity islands (PAIs and 10 virulence factors (VFs. E. coli strains were predominantly of the B1 and B2 phylotype, with few strains in the A or D phylotype. The PAIs IV536, ICFT073, and IICFT073 had the highest prevalence: 76%, 64%, and 48%, respectively. The PAIs II536, IJ96, and IIJ96 were less prevalent: 28%, 20%, and 24%, respectively. The most prevalent VF was vat (40%, while the least prevalent VFs were sfa (8% and iha (12%. None of the strains carried the VF fyuA, which is very common in uropathogenic E. coli (UPEC. The genetic profiles of E. coli in this cohort seem to be more similar to UPEC than to commensal E. coli. However, they appear to have reduced virulence potential that allows them to colonize asymptomatically.

  16. Multiple antimicrobial resistance among Avian Escherichia coli strains in Albania

    Directory of Open Access Journals (Sweden)

    Antonio Camarda

    2010-01-01

    Full Text Available In this study, 101 Escherichia (E. coli isolates from broilers, laying hens and turkeys which had died from colibacillosis, collected from 37 intensive and rural farms in Albania, were tested for antimicrobial susceptibility toward 12 different molecules. The highest levels of resistance were observed for Erythromycin (E (100% Amoxicillin (AMX (99.1%, Tetracycline (TE 30 (96.07%, Streptomycin (STR (93.07% and Neomycin (N30 (85.15%. Considerable resistance was also detected for fluoroquinolones. Moreover, 73.33% of E. coli resistant to at least one fluoroquinolone were also resistant to the two other fluoroquinolones checked. No evident differences were found between the E. coli from intensive and from rural farms. Multiple antibiotic resistance was expressed by all the E. coli tested. 23.63% and 17.39% of E. coli isolated from intensive and rural farms, respectively, were resistant towards all the drugs tested. These data would seem to indicate incorrect use of antibiotics on poultry farms in Albania.

  17. Risk factors for fecal colonization with multiple distinct strains of Escherichia coli among long-term care facility residents.

    Science.gov (United States)

    Lautenbach, Ebbing; Tolomeo, Pam; Black, Nicole; Maslow, Joel N

    2009-05-01

    Of 49 long-term care facility residents, 21 (43%) were colonized with 2 or more distinct strains of Escherichia coli. There were no significant risk factors for colonization with multiple strains of E. coli. These results suggest that future efforts to efficiently identify the diversity of colonizing strains will be challenging.

  18. Multi-omics Quantification of Species Variation of Escherichia coli Links Molecular Features with Strain Phenotypes

    DEFF Research Database (Denmark)

    Monk, Jonathan M; Koza, Anna; Campodonico, Miguel A;

    2016-01-01

    Escherichia coli strains are widely used in academic research and biotechnology. New technologies for quantifying strain-specific differences and their underlying contributing factors promise greater understanding of how these differences significantly impact physiology, synthetic biology......, metabolic engineering, and process design. Here, we quantified strain-specific differences in seven widely used strains of E. coli (BL21, C, Crooks, DH5a, K-12 MG1655, K-12 W3110, and W) using genomics, phenomics, transcriptomics, and genome-scale modeling. Metabolic physiology and gene expression varied......-specific caveats. Integrated modeling revealed that certain strains are better suited to produce given compounds or express desired constructs considering native expression states of pathways that enable high-production phenotypes. This study yields a framework for quantitatively comparing strains in a species...

  19. Accumulation of ciprofloxacin and lomefloxacinin fluoroquinolone-resistant strains of Escherichia coli

    Institute of Scientific and Technical Information of China (English)

    夏培元; 冯萍; 钟利; 吕晓菊; 雷秉钧

    2002-01-01

    Objective To evaluate the role of outer membrane protein (Omp) F-deficiency and active efflux in the accumulation of hydrophilic fluoroquinolones ciprofloxacin (CPLX) and lomefloxacin (LMLX) in resistant E. coli strains. Methods Fluoroquinolone accumulation in bacteria and the effect of active efflux were measured by a fluorescence method. The outer membrane proteins of the bacteria were analysed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). E. coli strains in this study included control strains JF701 and JF703 that are OmpC- or OmpF-deficient mutants of E. coli K-12, respectively, and the fluoroquinolone susceptible strain the fluoroquinolone susceptible strain of Escherichia coli (Ecs) and its in vitroselected resistant strains R2 and R256, and the clinical resistant isolates R5 and R6. Results The steady-state accumulation concentration of each drug in Ecs appeared to be the same as in JF701, while in the OmpF- deficient strain JF703, it was 1/5 CPLX or 1/2 LMLX lower than that in JF701, but JF703 was still susceptible to fluoroquinolones. On the other hand, compared with susceptible strains, a 2- to 10-fold decrease in the accumulation of each drug was found in the resistant strains except R2, in which the accumulation was slightly higher than in JF703. After the addition of 2,4-dinitrophenol (DNP), accumulation of each drug increased, especially in resistant strains, indicating that the function of the active efflux (pump) system in these bacteria had been enhanced dramatically. Furthermore, both OmpF and OmpC in Ecs, OmpF-deficiency in R2 and R256 and OmpC-deficiency in R5 and R6 were observed.Conclusion The decreased accumulation of hydrophilic fluoroquinolones in E. coli involved OmpF-deficiency and active efflux (pump), and the latter may be an important factor.

  20. High temporal variability in commensal Escherichia coli strain communities of a herbivorous marsupial.

    Science.gov (United States)

    Blyton, Michaela D J; Banks, Sam C; Peakall, Rod; Gordon, David M

    2013-08-01

    Although Escherichia coli is an important model organism for bacterial research, few studies have explored the nature of temporal variation in E. coli strains within the intestinal tracts of host individuals. In this study the E. coli strains of 54 mountain brushtail possums were sampled on four occasions during a year. This allowed temporal changes to be quantified both at the host population level and within individuals. Escherichia coli strains were identified using a combination of rep-PCR profiles from two primers (CGG and ERIC) and phylogenetic group assigned by quadruplex PCR. The study revealed considerable changes in community structure within individuals among all time periods. In fact, temporal variation within individuals accounted for more of the variation in E. coli community structure than differences between animals. In contrast to the within-host dynamics, there were no significant differences among the time periods at the host population level. It was also found that there was no effect of host age or sex on strain community structure within host individuals. These findings highlight the importance of temporal variation in the ecology of E. coli, while the methods applied in this study may serve as a foundation for further work in this area.

  1. [Antibiotic resistance of Escherichia coli strains isolated from raw chicken meat in Senegal].

    Science.gov (United States)

    Fofana, A; Bada Alambédji, R; Seydi, M; Akakpo, A J

    2006-01-01

    Antimicrobial-resistant Escherichia coli and others pathogens bacteria can be transferred from animals to humans through consumption of contaminated food and foods products and thus present a public health risk. The increase in E. coli resistance to commonly used antimicrobials both in the public health and veterinary sectors is one of the major threats of health care worldwide. The present study was undertaken to estimate the antimicrobial resistance of E. coli isolates from raw chicken meat in Dakar. Levying of skin and muscle have been carried out on 120 chicken carcasses bought from 13 sale points and 23 flocks beetween November 2003 and April 2004. 102 Escherichia coli strains have been isolated, among which, 90 were tested for their susceptibilities to 16 selected antibiotics by agar diffusion method. All Escherichia coli strains (100%), were resistant to one or more antibiotic; 60 strains (66.66%) being resistant to more than five antibiotics. Those frequently encountererd are: ampicillin, trimethoprim, trimethoprim-sulfametoxazole, tetracycline, sulfonamides, streptomycin, nalidixic acid. Multiple resistances to 12 antibiotics were also observed. The lowest resistances were noted with gentamicin (3.33%) and neomycin (5.56%). This study showed the significance of chicken meat as source of Escherichia coli strains with a simple or multiple resistance to various antibiotics tested. Further studies are necessary in order to determine bacterium mechanisms of resistance.

  2. [Antibioresistance of Escherichia coli strains isolated from raw chicken meat in Senegal].

    Science.gov (United States)

    Fofana, A; Bada Alambedji, R; Seydi, M; Akakpo, A J

    2006-01-01

    Antimicrobial-resistant Escherichia coli and others pathogens bacteria can be transferred from animals to humans through consumption of contaminated food and foods products and thus present a public health risk. The increase in E. coli resistance to commonly used antimicrobials both in the public health and veterinary sectors is one of the major threats of health care worldwide. The present study was undertaken to estimate the antimicrobial resistance of E. coli isolates from raw chicken meat in Dakar. Levying of skin and muscle have been carried out on 120 chicken carcasses bought from 13 sale points and 23 flocks beetween November 2003 and April 2004. 102 Escherichia coli strains have been isolated, among which, 90 were tested for their susceptibilities to 16 selected antibiotics by agar diffusion method. All Escherichia coli strains (100%), were resistant to one or more antibiotic; 60 strains (66.66%) being resistant to more than five antibiotics. Those frequently encountererd are: ampicillin, trimethoprim, trimethoprim-sulfametoxazole, tetracycline, sulfonamides, streptomycin, nalidixic acid. Multiple resistances to 12 antibiotics were also observed. The lowest resistances were noted with gentamicin (3.33%) and neomycin (5.56%). This study showed the significance of chicken meat as source of Escherichia coli strains with a simple or multiple resistance to various antibiotics tested. Further studies are necessary in order to determine bacterium mechanisms of resistance.

  3. Early settlers: which E. coli strains do you not want at birth?

    Science.gov (United States)

    Secher, Thomas; Brehin, Camille; Oswald, Eric

    2016-07-01

    The intestinal microbiota exerts vital biological processes throughout the human lifetime, and imbalances in its composition have been implicated in both health and disease status. Upon birth, the neonatal gut moves from a barely sterile to a massively colonized environment. The development of the intestinal microbiota during the first year of life is characterized by rapid and important changes in microbial composition, diversity, and magnitude. The pioneer bacteria colonizing the postnatal intestinal tract profoundly contribute to the establishment of the host-microbe symbiosis, which is essential for health throughout life. Escherichia coli is one of the first colonizers of the gut after birth. E. coli is a versatile population including harmless commensal, probiotic strains as well as frequently deadly pathogens. The prevalence of the specific phylogenetic B2 group, which encompasses both commensal and extra- or intraintestinal pathogenic E. coli strains, is increasing among E. coli strains colonizing infants quickly after birth. Fifty percent of the B2 group strains carry in their genome the pks gene cluster encoding the synthesis of a nonribosomal peptide-polyketide hybrid genotoxin named colibactin. In this review, we summarize both clinical and experimental evidence associating the recently emerging neonatal B2 E. coli population with several pathology and discuss how the expression of colibactin by both normal inhabitants of intestinal microflora and virulent strains may darken the borderline between commensalism and pathogenicity. Copyright © 2016 the American Physiological Society.

  4. Engineering of a plasmid-free Escherichia coli strain for improved in vivo biosynthesis of astaxanthin

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    Steuer Kristin

    2011-04-01

    Full Text Available Abstract Background The xanthophyll astaxanthin is a high-value compound with applications in the nutraceutical, cosmetic, food, and animal feed industries. Besides chemical synthesis and extraction from naturally producing organisms like Haematococcus pluvialis, heterologous biosynthesis in non-carotenogenic microorganisms like Escherichia coli, is a promising alternative for sustainable production of natural astaxanthin. Recent achievements in the metabolic engineering of E. coli strains have led to a significant increase in the productivity of carotenoids like lycopene or β-carotene by increasing the metabolic flux towards the isoprenoid precursors. For the heterologous biosynthesis of astaxanthin in E. coli, however, the conversion of β-carotene to astaxanthin is obviously the most critical step towards an efficient biosynthesis of astaxanthin. Results Here we report the construction of the first plasmid-free E. coli strain that produces astaxanthin as the sole carotenoid compound with a yield of 1.4 mg/g cdw (E. coli BW-ASTA. This engineered E. coli strain harbors xanthophyll biosynthetic genes from Pantoea ananatis and Nostoc punctiforme as individual expression cassettes on the chromosome and is based on a β-carotene-producing strain (E. coli BW-CARO recently developed in our lab. E. coli BW-CARO has an enhanced biosynthesis of the isoprenoid precursor isopentenyl diphosphate (IPP and produces β-carotene in a concentration of 6.2 mg/g cdw. The expression of crtEBIY along with the β-carotene-ketolase gene crtW148 (NpF4798 and the β-carotene-hydroxylase gene (crtZ under controlled expression conditions in E. coli BW-ASTA directed the pathway exclusively towards the desired product astaxanthin (1.4 mg/g cdw. Conclusions By using the λ-Red recombineering technique, genes encoding for the astaxanthin biosynthesis pathway were stably integrated into the chromosome of E. coli. The expression levels of chromosomal integrated recombinant

  5. Cytolethal distending toxin-producing Escherichia coli strains causing severe diarrhoea in young Mexican children

    Science.gov (United States)

    Maldonado-Puga, Samantha; Huerta-Cantillo, Jazmin; Chavez-Dueñas, Lucia; Navarro-Garcia, Fernando

    2017-01-01

    Introduction. Cytolethal distending toxins (CDTs), encoded by cdt genes, have DNase activity leading to cellular and nuclear distension, resulting in irreversible cell cycle arrest and apoptosis of target cells. cdt-positive Escherichia coli strains have been isolated from children with diarrhoea. There is, however, scant information on the prevalence and clinical presentation of diarrhoeal disease caused by these strains. Furthermore, toxin production of cdt-positive strains is rarely confirmed. We report five young children with diarrhoea caused by CDT-producing E. coli in whom stools were negative for other bacterial or enteric pathogens. Case presentation. On admission to hospital, all children presented watery diarrhoea with high stool output (range 7–20 stools/24 h); five had fever of 38 °C or more and four presented vomiting. Dehydration was present in four patients, one of whom had hypovolaemic shock; one child also presented hyponatraemia and hypokalaemia. In two children, cdt-positive strains were classified as typical and atypical enteropathogenic E. coli, and the remaining three harboured cdt-positive strains that did not belong to any diarrhoeagenic pathogroup. One cdt-positive strain from each case was characterized by a CDT cytotoxic assay and a cdt type-specific PCR. All strains produced the characteristic cellular intoxication due to CDT. Two strains carried the cdt-I, one cdt-III, one cdt-IV, and one concurrently had cdt-I, cdt-II and cdt-III genes. Conclusion. Our results suggest that CDT-producing E. coli strains are an infrequent, albeit significant, cause of severe diarrhoeal illness in children. Future research should measure the true burden of cdt-positive E. coli diarrhoea among children.

  6. Molecular and antimicrobial susceptibility analyses distinguish clinical from bovine Escherichia coli O157 strains.

    Science.gov (United States)

    Vidovic, Sinisa; Tsoi, Sarah; Medihala, Prabhakara; Liu, Juxin; Wylie, John L; Levett, Paul N; Korber, Darren R

    2013-07-01

    A population-based study combining (i) antimicrobial, (ii) genetic, and (iii) virulence analyses with molecular evolutionary analyses revealed segregative characteristics distinguishing human clinical and bovine Escherichia coli O157 strains from western Canada. Human (n = 50) and bovine (n = 50) strains of E. coli O157 were collected from Saskatchewan and Manitoba in 2006 and were analyzed by using the six-marker lineage-specific polymorphism assay (LSPA6), antimicrobial susceptibility analysis, the colicin assay, plasmid and virulence profiling including the eae, ehxA, espA, iha, stx1, stx2, stx2c, stx2d, stx2d-activatable, stx2e, and stx2f virulence-associated genes, and structure analyses. Multivariate logistic regression and Fisher's exact test strongly suggested that antimicrobial susceptibility was the most distinctive characteristic (P = 0.00487) associated with human strains. Among all genetic, virulence, and antimicrobial determinants, resistance to tetracycline (P coli O157 strains. Among 11 virulence-associated genes, stx2c showed the strongest association with E. coli O157 strains of bovine origin. LSPA6 genotyping showed the dominance of the lineage I genotype among clinical (90%) and bovine (70%) strains, indicating the importance of lineage I in O157 epidemiology and ecology. Population structure analysis revealed that the more-diverse bovine strains came from a unique group of strains characterized by a high degree of antimicrobial resistance and high frequencies of lineage II genotypes and stx2c variants. These findings imply that antimicrobial resistance generated among bovine strains of E. coli O157 has a large impact on the population of this human pathogen.

  7. Laboratory strains of Escherichia coli: model citizens or deceitful delinquents growing old disgracefully?

    Science.gov (United States)

    Hobman, Jon L; Penn, Charles W; Pallen, Mark J

    2007-05-01

    Escherichia coli stands unchallenged as biology's premier model organism. However, we propose, equipped with insights from the post-genomic era, a contrary view: that microbiology's chief idol has feet of clay. E. coli laboratory strains, particularly E. coli K-12, are far from model citizens, but instead degenerate and deceitful delinquents growing old disgracefully in our scientific institutions. E. coli K-12 is neither archetype nor ancestor. In addition, it has a far from optimal provenance for a model organism, with strong grounds for believing that current versions of the strain are quite distinct from any original wild-type free-living ancestor. In addition, it is usually studied under conditions far removed from its natural habitats and in ignorance of the selective pressures that have shaped its evolution. Fortunately, a flood of information from high-throughput genome sequencing, together with a new 'eco-evo' view of this model organism, promises to help put K-12 better into context.

  8. [An avian strain of Escherichia coli with antigens common to the genus Salmonella].

    Science.gov (United States)

    Terzolo, H R; Zoratti de Verona, A; d'Empaire, M; Furowicz, A J

    1977-01-01

    On a commercial poultry farm, a large percentage (9%) of clinically healthy fowls had positive reaction to the plate test, with commercial polyvalent pullorum antigens. We could not isolate Salmonella from the positive birds. An strain, of Escherichia coli Balcarce (E. coli B) was isolated from the feces of one of the birds. The isolate was identified biochemically and the antigenic study showed correlation with E. coli 044 and the somatic fraction 1, 2, 8, 14 and 23 of the Salmonella genus. The common antigens were studied by agglutination, absorption and crossed immunodiffusion tests, comparing the isolated strain and the different Salmonella serotypes. Four pullorum polyvalent commercial antigens reacted with sera containing somatic agglutinins 1, and with the E. coli B antiserum. These observations confirm the high antigenic correlation between the genus of the Enterobacteriaceae family. It is indicated that for the diagnosis of avian salmonelosis rather than using a single serological tests, the isolation and identification of the etiological agent is required.

  9. EMERGENCE OF MULTI DRUG RESISTANT STRAINS OF E. COLI ISOLATED FROM URINARY TRACT INFECTION IN NAMAKKAL

    Directory of Open Access Journals (Sweden)

    P. Poongothai

    2012-08-01

    Full Text Available In this study we evaluated the multi drug resistant strains of Escherichia coli isolated from urinary tract infection in different age groups in Namakkal. Totally 153 isolates of E.coli were obtained from 700 urine samples. Present study indicates that there is a high prevalence of multidrug resistant E. coli in female 24.34% followed by male 18.14%. After confirmation, isolates were performed to resistance patterns of different antibiotics were determined by standard disk diffusion method. The antibiogram patterns of the isolates showed a high percentage of multidrug resistance to cephodoxime, Novobiocin and vancomycin which was repeated to most of the isolates. Fifty resistance patterns were recognized among the MDR strains. This study revealed that E. coli was the predominant bacterial pathogen of community acquired UTIs in Aligarh, India. It also demonstrated an increasing resistance to number of antibiotics. This study is useful for clinician in order to improve the empirical treatment.

  10. Influence of subinhibitory concentrations of cefotaxime, imipenem and ciprofloxacin on adhesion of Escherichia coli strains to polystyrene.

    Science.gov (United States)

    Zalas-Wiecek, Patrycja; Gospodarek, Eugenia; Piecyk, Katarzyna

    2011-01-01

    The present study investigated the ability of sub MICs of cefotaxime, imipenem and ciprofloxacin to interfere with adhesion of E. coli strains to polystyrene (selected polymer used in studies on microorganisms' adhesion). It was observed that cefotaxime and imipenem at 1/2 and 1/4 MICs decreased the adherence of E. coli strains to polystyrene significantly. 1/2, 1/4 and 1/8 MICs of ciprofloxacin generally decreased the adhesive properties of E. coli strains, but two E. coli strains showed a noticeable enhancement of adhesion after incubation at sub MICs of this antibiotic.

  11. Diffusely adherent Escherichia coli strains isolated from children and adults constitute two different populations

    OpenAIRE

    Mansan-Almeida Rosane; Pereira Alex; Giugliano Loreny

    2013-01-01

    Abstract Background Diffusely adherent Escherichia coli (DAEC) have been considered a diarrheagenic category of E. coli for which several potential virulence factors have been described in the last few years. Despite this, epidemiological studies involving DAEC have shown inconsistent results. In this work, two different collections of DAEC possessing Afa/Dr genes, from children and adults, were studied regarding characteristics potentially associated to virulence. Results DAEC strains were r...

  12. "Emergence of Multidrug Resistant Strains of Escherichia coli Isolated from Urinary Tract Infections"

    OpenAIRE

    R Moniri; Khorshidi, A; H Akbari

    2003-01-01

    The emergence of multidrug resistant strains of Escherichia coli has complicated treatment decision and may lead to treatment failures. From April to November 2001 we prospectively evaluated the prevalence of resistance to trimethoprim-sulfamethoxazole (SXT), gentamicin, cephalothin, ciprofloxacin, and nitrofurantoin in 220 Escherichia coli isolates from patients with urinary tract infections in kashan, Iran. To assess the current breadth of multidrug resistance among urinary isolates of E. c...

  13. A Fatal Case of Necrotizing Fasciitis Caused by a Highly Virulent Escherichia coli Strain

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    Sadjia Bekal

    2016-01-01

    Full Text Available Necrotizing fasciitis is a serious disease characterized by the necrosis of the subcutaneous tissues and fascia. E. coli as the etiologic agent of necrotizing fasciitis is a rare occurrence. A 66-year-old woman underwent total abdominal hysterectomy with bilateral salpingo-oophorectomy. She rapidly developed necrotizing fasciitis which led to her death 68 hours following surgery. An E. coli strain was isolated from blood and fascia cultures. DNA microarray revealed the presence of 20 virulence genes.

  14. Comparison of the serum sensitivity of uropathogenic strains of Escherichia coli isolated from different diagnostic groups

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    J. Vraneš,

    2005-02-01

    Full Text Available The bactericidal activity of serum caused by complement system is an important defence mechanism protecting the host organism against infection. The capacity to resist bactericidal activity of normal human serum contributes to the virulence of many gram-negative pathogens. Serum resistance in bacteria has been attributed to their surface components, but exact mechanism of resistance which most likely involves multiple factors is not well understood. In this study, the capacity of Escherichia coli to resist the bactericidal action of serum was examined in 85 clinical isolates obtained from patients with acute pyelonephritis (n=23, acute cystitis (n=22, chronic pyelonephritis (n=22 and asymptomatic bacteriuria (n=18. Serum sensitivity was also examined in relation to the serogroup specificity and expression of the different adhesins of the strains.Bacterial susceptibility to serum killing was measured by assessing regrowth after incubation in serum according to Schiller and Hatch method. The adhesins of E. coli were determined by hemagglutination and inhibition of hemagglutiation, and serotyping was performed on glass slides and confirmed using a mechanized microtechnique.The significant correlation between serum resistance of uropathogenic strains of E. coli and expression of P-fimbriae and O6 serogroup was observed.Theincidence of serum-resistant E. coli strains was significantly higher in strains isolated from urine of patients with acute pyelonephritis, as compared to strains isolated in other diagnostic groups, which is in accordance with higher virulence and invasive potential of these strains.

  15. Antimicrobial Resistant Pattern of Escherichia Coli Strains Isolated from Pediatric Patients in Jordan

    Directory of Open Access Journals (Sweden)

    Mohammad Alshara

    2011-05-01

    Full Text Available The present study was conducted to investigate antimicrobial resistant pattern of Escherichia coli (E. coli strains isolated from clinical specimens of Jordanian pediatric patients during the period from January to December 2008. A total of 444 E. coli strains were isolated from clinical specimens and tested for their susceptibility to different antimicrobial drugs. Overall, high resistance rate was observed for ampicillin (84%, followed by amoxicillin-clavulanic acid (74.3%, cotrimoxazole (71%, nalidixic acid (47.3%, cephalothin (41%. Lower resistance rates were observed for amikacin (0% followed by Cefotaxime (11%, Ceftriaxone (11.7%, ciprofloxacin (14.5%, Norfloxacin (16.5%, gentamicin (17.3% cephalexin (20.9%, Ceftazidime (22.5%, cefixime (29.6%, and cefaclor (32.8%. Ampicillin, amoxicillin-clavulanic acid and cotrimoxazole were found to be ineffective at in vitro inhibition of the E. coli of pediatric origin. Amikacin was highly effective for E. coli with susceptibility rate of 100%. The majority of E. coli strains were susceptible to third generation cephalosporins and fluoroquinolones.

  16. PCR for the Specific Detection of an Escherichia coli O157:H7 Laboratory Control Strain.

    Science.gov (United States)

    Knowles, Michael; Lambert, Dominic; Huszczynski, George; Gauthier, Martine; Blais, Burton W

    2015-09-01

    Control strains of bacterial pathogens such as Escherichia coli O157:H7 are commonly processed in parallel with test samples in food microbiology laboratories as a quality control measure to assure the satisfactory performance of materials used in the analytical procedure. Before positive findings can be reported for risk management purposes, analysts must have a means of verifying that pathogenic bacteria (e.g., E. coli O157:H7) recovered from test samples are not due to inadvertent contamination with the control strain routinely handled in the laboratory environment. Here, we report on the application of an in-house bioinformatic pipeline for the identification of unique genomic signature sequences in the development of specific oligonucleotide primers enabling the identification of a common positive control strain, E. coli O157:H7 (ATCC 35150), using a simple PCR procedure.

  17. Targeted metabolic profiling rapidly differentiates Escherichia coli and Staphylococcus aureus at species and strain level.

    Science.gov (United States)

    Li, Haorong; Zhu, Jiangjiang

    2017-10-15

    Pathogenic foodborne bacteria have been associated with severe infectious disease in humans and animals worldwide. Rapid detection and screening of these foodborne pathogens are critical for our food safety. This study aimed at detecting Escherichia coli and Staphylococcus aureus, two important foodborne bacteria, at the species and strain/serovar level using a mass spectrometry (MS)-based targeted metabolic profiling approach. Ten E. coli strains (8 out of 10 were foodborne outbreak isolates) and four S. aureus strains were tested at two growth time points. A high-performance liquid chromatography/tandem mass spectrometry (HPLC/MS/MS)-based targeted metabolomics approach was applied for metabolic profile based bacteria detection. A total of 108 metabolites from multiple metabolic pathways were confidently detected from these bacteria. Our study demonstrated that with only 4 h of enrichment in the same medium, the metabolic profiles from E. coli and S. aureus showed significant difference. Furthermore, seven out of ten E. coli strains and all four tested S. aureus strains showed strain/serovar-level differentiation at the 4-h time point, which indicated great potential for strain level stratification in future food screening using our MS-based targeted metabolic profiling approach. A targeted metabolomics method was developed to demonstrate the utility of HPLC/MS/MS-based metabolic profiling in rapidly (4 h) differentiating E. coli and S. aureus bacteria, two of the most notorious foodborne bacteria, at both the species and strain/serovar levels. The results indicated that our approach has great potential in the future for fast and specific detection of foodborne pathogenic bacteria based on their metabolic diversity. Copyright © 2017 John Wiley & Sons, Ltd.

  18. Efficient recovery of fluoroquinolone-susceptible and fluoroquinolone-resistant Escherichia coli strains from frozen samples.

    Science.gov (United States)

    Lautenbach, Ebbing; Santana, Evelyn; Lee, Abby; Tolomeo, Pam; Black, Nicole; Babson, Andrew; Perencevich, Eli N; Harris, Anthony D; Smith, Catherine A; Maslow, Joel

    2008-04-01

    We assessed the rate of recovery of fluoroquinolone-resistant and fluoroquinolone-susceptible Escherichia coli isolates from culture of frozen perirectal swab samples compared with the results for culture of the same specimen before freezing. Recovery rates for these 2 classes of E. coli were 91% and 83%, respectively. The majority of distinct strains recovered from the initial sample were also recovered from the frozen sample. The strains that were not recovered were typically present only in low numbers in the initial sample. These findings emphasize the utility of frozen surveillance samples.

  19. Comparison of ruminant and human attaching and effacing Escherichia coli (AEEC) strains.

    Science.gov (United States)

    Horcajo, Pilar; Domínguez-Bernal, Gustavo; de la Fuente, Ricardo; Ruiz-Santa-Quiteria, José A; Blanco, Jesús E; Blanco, Miguel; Mora, Azucena; Dahbi, Ghizlane; López, Cecilia; Puentes, Beatriz; Alonso, María Pilar; Blanco, Jorge; Orden, José A

    2012-03-23

    The presence of 12 genes associated with virulence in human attaching and effacing Escherichia coli (AEEC) was studied within a collection of 20 enterohemorrhagic E. coli (EHEC) and 206 atypical enteropathogenic E. coli (EPEC) isolated from ruminants. In addition, virulence genes and the clonal relationship of 49 atypical EPEC O26 strains isolated from humans and ruminants were compared to clarify whether ruminants serve as a reservoir of atypical EPEC for humans. A great diversity in the content of virulence gene was found. Thus, the espH, espG and map genes were detected in more than 85% of ruminant AEEC strains; the tccP2, espI, efa1/lifA, ehxA and paa genes were present in 50-70% of strains; and other genes such as tccP, espP, katP and toxB were detected in <25% of strains. EHEC strains contained more virulence genes than atypical EPEC strains. Our results suggest for the first time that the efa1/lifA gene is associated with diarrhea in newborn ruminants and that the AEEC strains with the H11 flagellar antigen are potentially more virulent than the non-H11 AEEC strains. Importantly, we identified a new intimin variant gene, eaeρ, in three ruminant atypical EPEC strains. The comparison of ruminant and human EPEC O26 strains showed that some ruminant strains possess virulence gene profiles and pulse-field gel electrophoresis pulsotypes similar to those of human strains. In conclusion, our data suggest that atypical EPEC is a heterogeneous group with different pathogenic potential and that ruminants could serve as a reservoir of atypical EPEC for humans.

  20. Draft Genome Sequence of Escherichia coli 26R 793, a Plasmid-Free Recipient Strain Commonly Used in Conjugation Assays

    Science.gov (United States)

    Hurley, Daniel; McGrath, Kathleen; Bai, Li; Hächler, Herbert; Stephan, Roger

    2016-01-01

    Here, we report the draft genome sequence of the lactose-negative, rifampin-resistant, Escherichia coli strain 26R 793. This isolate has been widely used in conjugation experiments as a general recipient strain.

  1. Improvement of Escherichia coli production strains by modification of the phosphoenolpyruvate:sugar phosphotransferase system

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    Gosset Guillermo

    2005-05-01

    Full Text Available Abstract The application of metabolic engineering in Escherichia coli has resulted in the generation of strains with the capacity to produce metabolites of commercial interest. Biotechnological processes with these engineered strains frequently employ culture media containing glucose as the carbon and energy source. In E. coli, the phosphoenolpyruvate:sugar phosphotransferase system (PTS transports glucose when this sugar is present at concentrations like those used in production fermentations. This protein system is involved in phosphoenolpyruvate-dependent sugar transport, therefore, its activity has an important impact on carbon flux distribution in the phosphoenolpyruvate and pyruvate nodes. Furthermore, PTS has a very important role in carbon catabolite repression. The properties of PTS impose metabolic and regulatory constraints that can hinder strain productivity. For this reason, PTS has been a target for modification with the purpose of strain improvement. In this review, PTS characteristics most relevant to strain performance and the different strategies of PTS modification for strain improvement are discussed. Functional replacement of PTS by alternative phosphoenolpyruvate-independent uptake and phosphorylation activities has resulted in significant improvements in product yield from glucose and productivity for several classes of metabolites. In addition, inactivation of PTS components has been applied successfully as a strategy to abolish carbon catabolite repression, resulting in E. coli strains that use more efficiently sugar mixtures, such as those obtained from lignocellulosic hydrolysates.

  2. (13)C-metabolic flux analysis for mevalonate-producing strain of Escherichia coli.

    Science.gov (United States)

    Wada, Keisuke; Toya, Yoshihiro; Banno, Satomi; Yoshikawa, Katsunori; Matsuda, Fumio; Shimizu, Hiroshi

    2017-02-01

    Mevalonate (MVA) is used to produce various useful products such as drugs, cosmetics and food additives. An MVA-producing strain of Escherichia coli (engineered) was constructed by introducing mvaES genes from Enterococcus faecalis. The engineered strain produced 1.84 mmol/gDCW/h yielding 22% (C-mol/C-mol) of MVA from glucose in the aerobic exponential growth phase. The mass balance analysis revealed that the MVA yield of the engineered strain was close to the upper limit at the biomass yield. Since MVA is synthesized from acetyl-CoA using NADPH as a cofactor, the production of MVA affects central metabolism in terms of carbon utilization and NADPH requirements. The reason for this highly efficient MVA production was investigated based on (13)C-metabolic flux analysis. The estimated flux distributions revealed that the fluxes of acetate formation and the TCA cycle in the engineered strain were lower than those in the control strain. Although the oxidative pentose phosphate pathway is considered as the NADPH generating pathway in E. coli, no difference of the flux was observed between the control and engineered strains. The production/consumption balance of NADPH suggested that additional requirement of NADPH for MVA synthesis was obtained from the transhydrogenase reaction in the engineered strain. Comparison between the measured flux distribution and the ideal values for MVA production proposes a strategy for further engineering to improve the MVA production in E. coli.

  3. In vitro biofilm formation of commensal and pathogenic Escherichia coli strains: impact of environmental and genetic factors

    DEFF Research Database (Denmark)

    Reisner, A.; Krogfelt, Karen; Klein, B.M.

    2006-01-01

    Our understanding of Escherichia coli biofilm formation in vitro is based on studies of laboratory K-12 strains grown in standard media. However, pathogenic E. coli isolates differ substantially in their genetic repertoire from E. coli K-12 and are subject to heterogeneous environmental condition...

  4. Identification of IbeR as a Stationary-Phase Regulator in Meningitic Escherichia coli K1 that Carries a Loss-of-Function Mutation in rpoS

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    Feng Chi

    2009-01-01

    Full Text Available IbeR is a regulator present in meningitic Escherichia coli strain E44 that carries a loss-of-function mutation in the stationary-phase (SP regulatory gene rpoS. In order to determine whether IbeR is an SP regulator in E44, two-dimensional gel electrophoresis and LC-MS were used to compare the proteomes of a noninvasive ibeR deletion mutant BR2 and its parent strain E44 in the SP. Four up-regulated (TufB, GapA, OmpA, AhpC and three down-regulated (LpdA, TnaA, OpmC proteins in BR2 were identified when compared to E44. All these proteins contribute to energy metabolism or stress resistance, which is related to SP regulation. One of the down-regulated proteins, tryptophanase (TnaA, which is regulated by RpoS in other E. coli strains, is associated with SP regulation via production of a signal molecule indole. Our studies demonstrated that TnaA was required for E44 invasion, and that indole was able to restore the noninvasive phenotype of the tnaA mutant. The production of indole was significantly reduced in BR2, indicating that ibeR is required for the indole production via tnaA. Survival studies under different stress conditions indicated that IbeR contributed to bacteria stress resistance in the SP. Taken together, IbeR is a novel regulator contributing to the SP regulation.

  5. Large scale analysis of virulence genes in Escherichia coli strains isolated from Avalon Bay, CA.

    Science.gov (United States)

    Hamilton, Matthew J; Hadi, Asbah Z; Griffith, John F; Ishii, Satoshi; Sadowsky, Michael J

    2010-10-01

    Contamination of recreational waters with Escherichia coli and Enterococcus sp. is a widespread problem resulting in beach closures and loss of recreational activity. While E. coli is frequently used as an indicator of fecal contamination, and has been extensively measured in waterways, few studies have examined the presence of potentially pathogenic E. coli strains in beach waters. In this study, a combination of high-throughput, robot-assisted colony hybridization and PCR-based analyses were used to determine the genomic composition and frequency of virulence genes present in E. coli isolated from beach water in Avalon Bay, Santa Catalina Island, CA. A total of 24,493 E. coli isolates were collected from two sites at a popular swimming beach between August through September 2007 and from July through August 2008. All isolates were examined for the presence of shiga-like toxins (stx1/stx2), intimin (eaeA), and enterotoxins (ST/LT). Of the 24,493 isolates examined, 3.6% contained the eaeA gene, indicating that these isolates were potential EPEC strains. On five dates, however, greater than 10% of the strains were potential EPEC, suggesting that incidence of virulence genes at this beach has a strong temporal component. No STEC or ETEC isolates were detected, and only eight (water and their presence needs to be considered as one of the factors used in decisions concerning beach closures.

  6. Diversity of Mercury Resistant Escherichia coli Strains Isolated from Aquatic Systems in Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    Raquel Costa de Luca Rebello

    2013-01-01

    Full Text Available Escherichia coli may harbor genetic mercury resistance markers which makes this bacterial species a promising alternative for bioremediation processes. The objective of this study was to investigate phenotypic and genetic characteristics related to diversity and mercury resistance among 178 Escherichia coli strains isolated from residential, industrial, agricultural, and hospital wastewaters and recreational waters at Rio de Janeiro city. Genetic and conventional methods were carried out in order to determine mercury resistance. Random amplification of polymorphic DNA (RAPD-PCR and denaturing gradient gel electrophoresis (DGGE were used to investigate genetic variability. RAPD data revealed a high degree of polymorphism among E. coli mercury resistant strains and showed reproducibility and good discriminative results. DGGE typing detected diversity within the merA gene fragment. Our findings represent an improvement in epidemiological studies of HgR  E. coli and support the evidence of nonclonal nature of mercury resistant E. coli strains circulating in rural and urban aquatic systems in Rio de Janeiro city.

  7. Isolation of a carbapenem-resistant K1 serotype Klebsiella pneumonia strain and the study of resistance mechanism%碳青霉烯类抗生素耐药K1型肺炎克雷伯菌一株的分离及耐药机制分析

    Institute of Scientific and Technical Information of China (English)

    张嵘; 王选; 吕建新

    2014-01-01

    目的 对临床分离的1株碳青霉烯类抗生素耐药K1型肺炎克雷伯菌进行毒力及耐药机制分析.方法 采用Vitek 2 Compact全自动微生物鉴定仪对细菌进行鉴定;浓度梯度法(E-test)测定细菌对临床常用抗生素的最低抑菌浓度(MIC);乙二胺四乙酸(EDTA)协同试验和改良Hodge试验分别检测金属酶和碳青霉烯酶;聚合酶链反应(PCR)扩增毒力基因K1、K2、K5、K20、K54、K57、rmpA、magA、wcaG和β内酰胺类抗生素耐药基因;接合试验检测耐药基因和毒力基因是否存在于质粒上;接合子质粒进行PBRT质粒分型;多位点序列分析(MLST)对该菌株进行分子分型.结果 药敏结果显示该菌株对临床常用的头孢类和碳青霉烯类抗生素耐药;EDTA协同试验和改良Hodge试验显示该菌株产碳青霉烯酶,不含金属酶;PCR扩增及序列分析发现该菌株携带耐药基因blaKPC-2、blaCTX-M-15、blaTEM-1、blaSHV-1,同时有毒力基因K1、magA、rmpA、wcaG;能通过接合试验将碳青霉烯耐药基因传递到大肠埃希菌EC600,接合子未发现毒力基因存在;PBRT质粒分型为Frep型质粒;MLST分型为ST23型.结论 产毒力基因的K1型肺炎克雷伯菌携带blaKPC-2碳青霉烯耐药基因,该耐药基因可以通过质粒发生转移.%Objective To study the virulence and mechanism of carbapenem resistance of a clinical isolate of carbapenem-resistant K1 serotype Klebsiella pneumonia strain.Methods Identification of isolate was carried out with VITEK-2 compact system.Antimicrobial susceptibility was determined by E-test; Metallo β-lactamases and carbapenemases screening were conducted by imipenem-EDTA double disc synergy test and modified Hodge test,respectively.Specific polymerehse chain reaction (PCR) and DNA sequencing were preformed to detect the virulence genes including K1,K2,KS,K20,K54,K57,magA,rmpA,wcaG and a series of β-lactamase resistence genes.Conjunction experiment was also performed.The plasmids of

  8. Biochemical aspects of the resistance to nourseothricin (streptothricin) of Escherichia coli strains.

    Science.gov (United States)

    Seltmann, G

    1989-01-01

    In most cases Escherichia coli strains phenotypically resistant against nourseothricin (streptothricin) harbour a plasmid which codes for an acetyltransferase. This enzyme transfers an acetyl group from acetyl-coenzyme A to an amino group of the beta-lysine (peptide) chain of the antibiotic, thus inactivating it. Additionally, the penetrability for nourseothricin of the cell wall is drastically reduced in a high percentage of the resistant strains. Both resistance mechanisms seem to be independent of each other.

  9. Metaproteomics analyses as diagnostic tool for differentiation of Escherichia coli strains in outbreaks

    Science.gov (United States)

    Jabbour, Rabih E.; Wright, James D.; Deshpande, Samir V.; Wade, Mary; McCubbin, Patrick; Bevilacqua, Vicky

    2013-05-01

    The secreted proteins of the enterohemorrhagic and enteropathogenic E. coli (EHEC and EPEC) are the most common cause of hemorrhagic colitis, a bloody diarrhea with EHEC infection, which often can lead to life threatening hemolytic-uremic syndrome (HUS).We are employing a metaproteomic approach as an effective and complimentary technique to the current genomic based approaches. This metaproteomic approach will evaluate the secreted proteins associated with pathogenicity and utilize their signatures as differentiation biomarkers between EHEC and EPEC strains. The result showed that the identified tryptic peptides of the secreted proteins extracted from different EHEC and EPEC growths have difference in their amino acids sequences and could potentially utilized as biomarkers for the studied E. coli strains. Analysis of extract from EHEC O104:H4 resulted in identification of a multidrug efflux protein, which belongs to the family of fusion proteins that are responsible of cell transportation. Experimental peptides identified lies in the region of the HlyD haemolysin secretion protein-D that is responsible for transporting the haemolysin A toxin. Moreover, the taxonomic classification of EHEC O104:H4 showed closest match with E. coli E55989, which is in agreement with genomic sequencing studies that were done extensively on the mentioned strain. The taxonomic results showed strain level classification for the studied strains and distinctive separation among the strains. Comparative proteomic calculations showed separation between EHEC O157:H7 and O104:H4 in replicate samples using cluster analysis. There are no reported studies addressing the characterization of secreted proteins in various enhanced growth media and utilizing them as biomarkers for strain differentiation. The results of FY-2012 are promising to pursue further experimentation to statistically validate the results and to further explore the impact of environmental conditions on the nature of the secreted

  10. SIMULTANEOUS EFFECTS OF SHAKING AND TEMPERATURE ON VEROTOXIN1 PHAGE INDUCTION FROM VEROTOXIGENIC ESCHERICHIA COLI STRAINS

    Directory of Open Access Journals (Sweden)

    H. Hosain Zadegan, M. Sattari, M. H. Zahir, A. A. Allame

    2006-01-01

    Full Text Available Induction of lambda phage carring verotoxin1 gene from a verotoxigenic strains of Escherichia coli and released verotoxin1 were studied under environmental factors of shaking and termperature. Verotoxin1 phage in Escherichia coli PA 101 and transductants was confirmed by bacteriophage detection assay. Shaking of culture media and increasing temperature until 42 ºC increased phage particles in supernatants of Escherichia coli PA 101. Our results indicate that environmental factors such as shaking movements in natural inhabitates of bacteria such as river or sewage streams and temperature rise in summer season could be factors in induce and release free verotoxin1 – producing phage particles in nature that in turn could be the source of phage spreading to other related bacteria , and responsible for increased outbreaks of food borne diseases with verotoxigenic Escherichia coli in warm monthes of year in tropical areas.

  11. Fatal necrotizing fasciitis due to necrotic toxin-producing Escherichia coli strain

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    C. Gallois

    2015-11-01

    Full Text Available We report a fatal case of necrotizing soft tissues infection caused by an Escherichia coli strain belonging to phylogenetic group C and harbouring numerous virulence factors reported to be part of a pathogenicity island (PAI such as PAI IIJ96 and conserved virulence plasmidic region.

  12. The complete genome sequences of 65 Campylobacter jejuni and C. coli strains

    Science.gov (United States)

    Campylobacter jejuni (Cj) and C. coli (Cc) are genetically highly diverse based on various molecular methods including MLST, microarray-based comparisons and the whole genome sequences of a few strains. Cj and Cc diversity is also exhibited by variable capsular polysaccharides (CPS) that are the maj...

  13. Fatal necrotizing fasciitis due to necrotic toxin-producing Escherichia coli strain

    Science.gov (United States)

    Gallois, C.; Hauw-Berlemont, C.; Richaud, C.; Bonacorsi, S.; Diehl, J.-L.; Mainardi, J.-L.

    2015-01-01

    We report a fatal case of necrotizing soft tissues infection caused by an Escherichia coli strain belonging to phylogenetic group C and harbouring numerous virulence factors reported to be part of a pathogenicity island (PAI) such as PAI IIJ96 and conserved virulence plasmidic region. PMID:26693024

  14. Markerless Escherichia coli rrn Deletion Strains for Genetic Determination of Ribosomal Binding Sites

    DEFF Research Database (Denmark)

    Quan, Selwyn; Skovgaard, Ole; McLaughlin, Robert E

    2015-01-01

    Single-copy rrn strains facilitate genetic ribosomal studies in Escherichia coli. Consecutive markerless deletion of rrn operons resulted in slower growth upon inactivation of the fourth copy, which was reversed by supplying transfer RNA genes encoded in rrn operons in trans. Removal of the sixth...

  15. Development of Cultivation Technology for the Escherichia coli Recombinant Strain Producing Argininedeiminase of Mycoplasma hominis

    OpenAIRE

    Fayura, L R; Boretsky, Yu.R.; Pynyaha, Yu.V.; Martynyuk, N.B.; Skorohod, V.V.; Sybyrny, А.А.

    2014-01-01

    The recombinant Escherichia coli strain producing arginine deiminase of Mycoplasma hominis has been constructed. Storage conditions that provide stabilization of most productive clones of the producer were found. Terms for cultivation of the arginine deiminase producer using bioreactors of different volume were optimized. Highly purified samples of arginine deiminase were obtained and their longterm storage conditions were selected.

  16. Development of Cultivation Technology for the Escherichia coli Recombinant Strain Producing Argininedeiminase of Mycoplasma hominis

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    Fayura, L.R.

    2014-07-01

    Full Text Available The recombinant Escherichia coli strain producing arginine deiminase of Mycoplasma hominis has been constructed. Storage conditions that provide stabilization of most productive clones of the producer were found. Terms for cultivation of the arginine deiminase producer using bioreactors of different volume were optimized. Highly purified samples of arginine deiminase were obtained and their longterm storage conditions were selected.

  17. Innate immune evasion of Escherichia coli clinical strains from orthopedic implant infections.

    Science.gov (United States)

    Crémet, L; Broquet, A; Jacqueline, C; Chaillou, C; Asehnoune, K; Corvec, S; Caroff, N

    2016-06-01

    Escherichia coli is one of the first causes of Gram-negative orthopedic implant infections (OII). Those infections, usually hematogenous, mostly originate from the urinary tract. We investigated the strategies developed by E. coli in this context to evade host innate immune responses, i.e. complement and polymorphonuclear neutrophils (PMN). Twenty strains from OII were compared with 20 strains from bacteremia in patients with non-infected orthopedic implant. In both groups, 6/20 (30 %) strains lysed PMNs, due to the production of the pore-forming toxin α-hemolysin (HlyA). For the others, resistance to phagocytic killing by PMN was not significantly different between both groups. In contrast, resistance to complement-mediated serum killing was significantly higher in OII strains than in the others (65 % vs 10 %; P coli, different mechanisms have been involved in complement resistance. Here, serum resistance was not linked to a group 2 capsule, or a loss of outer membrane permeability, or the recruitment of the complement inhibitor C4bp, but was significantly associated with the synthesis of long-chain LPS, regardless of the O-antigen. Thus, serum resistance could promote seeding of peri-implant tissues by helping E. coli to either persist in blood and reach the site of infection or overcome localized complement activation.

  18. Multidrug resistant Escherichia coli strains isolated from urine sample, University of Gondar Hospital, Northwest Ethiopia

    Institute of Scientific and Technical Information of China (English)

    Setegn Eshetie; Fentahun Tarekegn; Gemechu Kumera; Feleke Mekonnen

    2016-01-01

    Objective: To assess multidrug resistant (MDR) Escherichia coli (E. coli) isolates from patients with urinary tract infection. Methods: From February to June 2014, a cross sectional study was conducted among urinary tract infection patients at the University of Gondar Hospital. Culture and disk diffusion method were used for E. coli isolation and to determine the antibiotic susceptibility patterns. Data were entered and analyzed using SPSS version 20. P Results: A total of 112 E. coli isolates were identified and the rate of isolation was higher among female participants (28.7%; P = 0.03). Of the isolates, 104 (92.9%) were MDR E. coli; and the isolates showed high resistance rates towards ampicillin (99%), cotrimoxazole (69%), chloramphenicol (58.7%), gentamycin (56.7%) and ceftazidime (55.8%). However, comparative isolates showed low resistance rates to ciprofloxacin (1%), cefepime (8.7%), and ceftriaxone (11.5%). Moreover, resistance rates of MDR E. coli isolates were significantly higher than non-MDR strains for ceftazidime (55.8% versus 12.5%; P = 0.015), and ampicillin (99% versus 87.5%; P = 0.018). Conclusions: High prevalence of MDR E. coli isolates was observed in this study. Regular monitoring of antibiotic resistance rates is necessarily required to improve and revise empirical antibiotic therapy protocols.

  19. Measuring and modelling straining of Escherichia coli in saturated porous media.

    Science.gov (United States)

    Foppen, Jan Willem; van Herwerden, Manon; Schijven, Jack

    2007-08-15

    Though coliform bacteria are used worldwide to indicate fecal pollution of groundwater, the parameters determining the transport of Escherichia coli in aquifers are relatively unknown. We evaluated the occurrence of both straining and attachment of E. coli ATCC25922 in columns of ultra-pure, angular, saturated quartz sand. The column experiments were conducted over a wide range of porous medium sizes, column heights, input concentrations, and pore water flow velocities. Straining and attachment were examined by modelling the breakthrough curves (with HYDRUS 1D). In addition, model output was compared with measured strained and attached bacteria via column extrusion experiments (in which sand was extruded from the column and placed in excess water) and flow reversal experiments (in which the pore water flow direction was reversed, thereby dislodging strained bacteria). Our model consisted of an attachment rate coefficient and a straining rate coefficient; both of these decreased with transport distance. The straining rate coefficient also decreased in a Langmuirian way, in response to the filling of available pore space, which in turn depended on influent bacteria concentration, quartz grain diameter, and transport distance. The maximum strained fraction was 25-30% of total bacteria mass applied to the column; the maximum attached fraction was 30-35%. The fit between modelled and measured (strained and attached) bacteria masses was acceptable, as was the sensitivity of the model output to fitted parameter values. Our results lead to a new description for the time-dependent mass balance of strained bacteria, which entails using three fitting parameters. The results also imply that column experiments in combination with retention profiles (or various column lengths) are not enough to explain the retention processes in a column. Column extrusion and flow reversal experiments provide vital additional information on the occurrence and magnitude of straining. Our

  20. Pathogenicity island sequences of pyelonephritogenic Escherichia coli CFT073 are associated with virulent uropathogenic strains.

    Science.gov (United States)

    Kao, J S; Stucker, D M; Warren, J W; Mobley, H L

    1997-07-01

    Urinary tract infection is the most frequently diagnosed kidney and urologic disease, and Escherichia coli is by far the most common etiologic agent. Defined blocks of DNA termed pathogenicity islands have been found in uropathogenic strains to carry genes not generally found in fecal strains. We have identified one of these regions of DNA within the chromosome of the highly virulent E. coli CFT073, isolated from the blood and urine of a woman with acute pyelonephritis. This strain, which is cytotoxic for cultured renal cells and causes acute pyelonephritis in transurethrally infected CBA mice, contains two distinct copies of the pap operon and is hemolytic. One pap operon was localized on a cosmid clone which was used to identify three overlapping cosmid clones. By using restriction mapping, DNA hybridization, sequencing, and PCR amplification, a region of approximately 50 kb was found to be present in this uropathogenic strain and to have no corresponding sequences in E. coli K-12. This gene block also carries hemolysin genes hlyCABD. The pathogenicity island begins 7 bp downstream of dadX (catabolic alanine racemase; 26.55 min) and ends at a position in the K-12 genome 75 bp downstream of the metV tRNA gene (62.74 min); this suggests that a chromosomal rearrangement has occurred relative to the K-12 linkage map. The junctions of the pathogenicity island were verified by PCR amplification directly from the genomic DNA of strain CFT073. DNA sequencing within the boundaries of the junctions revealed genes not previously identified in E. coli or in some cases bearing no known homologs. When used as probes for DNA hybridization, these sequences were found significantly more often in strains associated with the clinical syndromes of cystitis (82%) and acute pyelonephritis (79%) than in fecal strains (19%; P < 0.001).

  1. Frequency of pap and pil operons in Escherichia coli strains associated with urinary infections.

    Science.gov (United States)

    Perugini, M R; Vidotto, M C

    1996-03-01

    Strains of E. coli isolated from patients with urinary tract infection were examined for P and type 1 adhesin production by colony hybridization with pap and pil operons. The P pili probe detected 45 (46.4%) of the total of 97 strains studied and the type 1 pili probe detected 83 (85.6%). The pap operon was detected in 39 (53.4%) of 73 strains isolated from urine of patients with urinary disease and in 6 (25.0%) of 24 strains isolated from feces of healthy individuals employed as controls (P = 0.029), and the pil operon was detected in 67 (91.8%) of the urinary strains and in 16 (66.6%) of the fecal strains (P = 0.007). Our data did not show significant differences in frequency of P pili among isolates from pyelonephritis (78.5%), cystitis (45.8%) and asymptomatic bacteriuria (54.5%). Type 1 pili were not associated with the different types of infection; the frequency of these pili was 100% in pyelonephritis and in asymptomatic bacteriuria, and 87.5% in cystitis. The incidence of pap operon in strains isolated from pyelonephritis and from asymptomatic bacteriuria was higher in 11- to 40-year old women. These data show a high frequency of pap and pil operons among uropathogenic strains of E. coli, which seems to be an important factor in the development of urinary infection.

  2. A comparison of Shiga-toxin 2 bacteriophage from classical enterohemorrhagic Escherichia coli serotypes and the German E. coli O104:H4 outbreak strain.

    Directory of Open Access Journals (Sweden)

    Chad R Laing

    Full Text Available Escherichia coli O104:H4 was associated with a severe foodborne disease outbreak originating in Germany in May 2011. More than 4000 illnesses and 50 deaths were reported. The outbreak strain was a typical enteroaggregative E. coli (EAEC that acquired an antibiotic resistance plasmid and a Shiga-toxin 2 (Stx2-encoding bacteriophage. Based on whole-genome phylogenies, the O104:H4 strain was most closely related to other EAEC strains; however, Stx2-bacteriophage are mobile, and do not necessarily share an evolutionary history with their bacterial host. In this study, we analyzed Stx2-bacteriophage from the E. coli O104:H4 outbreak isolates and compared them to all available Stx2-bacteriophage sequences. We also compared Stx2 production by an E. coli O104:H4 outbreak-associated isolate (ON-2011 to that of E. coli O157:H7 strains EDL933 and Sakai. Among the E. coli Stx2-phage sequences studied, that from O111:H- strain JB1-95 was most closely related phylogenetically to the Stx2-phage from the O104:H4 outbreak isolates. The phylogeny of most other Stx2-phage was largely concordant with their bacterial host genomes. Finally, O104:H4 strain ON-2011 produced less Stx2 than E. coli O157:H7 strains EDL933 and Sakai in culture; however, when mitomycin C was added, ON-2011 produced significantly more toxin than the E. coli O157:H7 strains. The Stx2-phage from the E. coli O104:H4 outbreak strain and the Stx2-phage from O111:H- strain JB1-95 likely share a common ancestor. Incongruence between the phylogenies of the Stx2-phage and their host genomes suggest the recent Stx2-phage acquisition by E. coli O104:H4. The increase in Stx2-production by ON-2011 following mitomycin C treatment may or may not be related to the high rates of hemolytic uremic syndrome associated with the German outbreak strain. Further studies are required to determine whether the elevated Stx2-production levels are due to bacteriophage or E. coli O104:H4 host related factors.

  3. Improving Engineered Escherichia coli strains for High-level Biosynthesis of Isobutyrate

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    Mingyong Xiong

    2015-05-01

    Full Text Available Isobutyrate is an important platform chemical with various industrial applications. Previously, a synthetic metabolic pathway was constructed in E. coli to produce isobutyrate from glucose. However, isobutanol was found to be a major byproduct. Herein, gene knockouts and enzyme overexpressions were performed to optimize further the engineered E. coli strain. Besides yqhD, the knockouts of three genes eutG, yiaY and ygjB increased isobutyrate production in shake flasks. Furthermore, the introduction of an additional padA on a medium copy number plasmid under the constitutive promoter significantly reduced isobutanol formation. The IBA15-2C strain (BW25113, DyqhD, DygjB; carrying two copies of padA produced 39.2% more isobutyrate (0.39 g/glucose yield, 80% of the theoretical maximum yield than IBA1-1C strain (BW25113, DyqhD; carrying one copy of padA. A scale-up process was also investigated for IBA15-2C strain to optimize the conditions for the production of isobutyrate in the fermentor. With Ca(OH2 as the base for pH control and 10% dissolved oxygen level, IBA15-2C strain produced 90 g/L isobutyrate after 144 h. This study has engineered E. coli to achieve biosynthesis of a nonnative compound with the highest titer and opened up the possibility of the industrial production of isobutyrate.

  4. A simple and effective method for construction of Escherichia coli strains proficient for genome engineering.

    Science.gov (United States)

    Ryu, Young Shin; Biswas, Rajesh Kumar; Shin, Kwangsu; Parisutham, Vinuselvi; Kim, Suk Min; Lee, Sung Kuk

    2014-01-01

    Multiplex genome engineering is a standalone recombineering tool for large-scale programming and accelerated evolution of cells. However, this advanced genome engineering technique has been limited to use in selected bacterial strains. We developed a simple and effective strain-independent method for effective genome engineering in Escherichia coli. The method involves introducing a suicide plasmid carrying the λ Red recombination system into the mutS gene. The suicide plasmid can be excised from the chromosome via selection in the absence of antibiotics, thus allowing transient inactivation of the mismatch repair system during genome engineering. In addition, we developed another suicide plasmid that enables integration of large DNA fragments into the lacZ genomic locus. These features enable this system to be applied in the exploitation of the benefits of genome engineering in synthetic biology, as well as the metabolic engineering of different strains of E. coli.

  5. A simple and effective method for construction of Escherichia coli strains proficient for genome engineering.

    Directory of Open Access Journals (Sweden)

    Young Shin Ryu

    Full Text Available Multiplex genome engineering is a standalone recombineering tool for large-scale programming and accelerated evolution of cells. However, this advanced genome engineering technique has been limited to use in selected bacterial strains. We developed a simple and effective strain-independent method for effective genome engineering in Escherichia coli. The method involves introducing a suicide plasmid carrying the λ Red recombination system into the mutS gene. The suicide plasmid can be excised from the chromosome via selection in the absence of antibiotics, thus allowing transient inactivation of the mismatch repair system during genome engineering. In addition, we developed another suicide plasmid that enables integration of large DNA fragments into the lacZ genomic locus. These features enable this system to be applied in the exploitation of the benefits of genome engineering in synthetic biology, as well as the metabolic engineering of different strains of E. coli.

  6. Detection of diarrheagenic Escherichia coli strains isolated from dogs and cats in Brazil.

    Science.gov (United States)

    Puño-Sarmiento, Juan; Medeiros, Leonardo; Chiconi, Carolina; Martins, Fernando; Pelayo, Jacinta; Rocha, Sérgio; Blanco, Jorge; Blanco, Miguel; Zanutto, Marcelo; Kobayashi, Renata; Nakazato, Gerson

    2013-10-25

    Escherichia coli are gut microbiota bacteria that can cause disease in some humans and other animals, including dogs and cats that humans often keep as pets. Diarrheagenic E. coli (DEC) strains are classified into six categories: enteropathogenic (EPEC), enterotoxigenic (ETEC), Shiga toxin-producing (STEC), enteroinvasive (EIEC), enteroaggregative (EAEC), and diffuse-adhering E. coli (DAEC). In this study 144 and 163 E. coli colonies were isolated from the fecal samples of 50 dogs and 50 cats, respectively, with and without diarrhea from a Veterinary Hospital (clinical isolates). The virulence factors were determined using multiplex Polymerase Chain Reaction. Adherence assays, antibacterial susceptibility and serotyping (somatic or flagellar antigens) were performed on DEC isolates. We found 25 (17.4%) and 4 (2.5%) DEC strains isolated from dogs and cats, respectively. Only the EPEC and EAEC pathotypes were found in both animals. Meanwhile, genes from other pathotypes (STEC, EIEC, and ETEC) were not found in these clinical isolates. All of the DEC strains showed mannose-resistant adherence to HEp-2 and HeLa cells, and aggregative adherence was predominant in these isolates. Multiresistant strains to antimicrobials were found in most DEC strains including usual and unusual antimicrobials in veterinary practices. The serotypes of these DEC isolates were variable. The ONT serotype was predominant in these isolates. Some serotypes found in our study were described to human DEC. Here, we demonstrate that pets carry virulent DEC genes, which are mainly strains of EPECs and EAECs. The presence of these virulence factors in isolates from animals without diarrhea suggests that pets can act as a reservoir for human infection.

  7. The SOS response is permitted in Escherichia coli strains deficient in the expression of the mazEF pathway.

    Science.gov (United States)

    Kalderon, Ziva; Kumar, Sathish; Engelberg-Kulka, Hanna

    2014-01-01

    The Escherichia coli (E. coli) SOS response is the largest, most complex, and best characterized bacterial network induced by DNA damage. It is controlled by a complex network involving the RecA and LexA proteins. We have previously shown that the SOS response to DNA damage is inhibited by various elements involved in the expression of the E. coli toxin-antitoxin mazEF pathway. Since the mazEF module is present on the chromosomes of most E. coli strains, here we asked: Why is the SOS response found in so many E. coli strains? Is the mazEF module present but inactive in those strains? We examined three E. coli strains used for studies of the SOS response, strains AB1932, BW25113, and MG1655. We found that each of these strains is either missing or inhibiting one of several elements involved in the expression of the mazEF-mediated death pathway. Thus, the SOS response only takes place in E. coli cells in which one or more elements of the E. coli toxin-antitoxin module mazEF or its downstream pathway is not functioning.

  8. Host Response to Porcine Strains of Escherichia coli in a Novel Pyelonephritis Model

    DEFF Research Database (Denmark)

    Isling, L. K.; Aalbæk, B.; Birck, M. M.

    2011-01-01

    The initial pathology and pathogenesis of pyelonephritis and the influence of different strains of Escherichia coli were investigated in a novel porcine model. Nine female pigs were divided into three groups (A, B and C) and inoculated repeatedly into one renal pelvis with porcine pyelonephritis E...... kidneys. Gross and microscopical lesions of acute pyelonephritis were demonstrated in all but one kidney inoculated with E. coli, but in none of the control kidneys. Renal parenchymal infiltration with both neutrophils and mononuclear cells, primarily CD3+ T lymphocytes, was observed at 6hpi. Most T...

  9. Sensitivity of pathogenic and attenuated E. coli O157:H7 strains to ultraviolet-C light as assessed by conventional plating methods and ethidium monoazide-PCR

    Science.gov (United States)

    In this study, the UV-C sensitivity of six pathogenic E. coli O157:H7 strains associated with recent outbreaks of foodborne illnesses and four attenuated E. coli O157:H7 strains was investigated. Futhermore, the mechanism of UV-C impact on two pathogenic E. coli strains with different UV-C sensitiv...

  10. [Study of phenotypical and antimicrobial susceptibility markers in enteric Escherichia coli strains].

    Science.gov (United States)

    Aguila, Adalberto; Bernedo, Robert; Llop, Alina; Ramírez, Margarita; Bravo, Laura; Fernández, Anabel; Ledo, Yudith

    2007-01-01

    Forty strains of Escherichia coli isolated from children under 5 years of age with acute diarreas, coming from different provinces of the country , were analyzed. Four important phenotypical determinants were tested: sorbosa, sorbitol, enterohemolysin and 0157:H7 serology, in order to select those strains from enterohemorrhagic or Shiga toxin-producing category. Likewise, they were characterized by biotyping and antimicrobial susceptibility methods. The use of phenotypical tests showed six strains with presumptive characteristics, four of which were most likely to be Shiga toxin-producing strains. In antimicrobial susceptibility test, the strains showed high resistance mainly to ampicillin and trimethrophin-sulfamethoxasole. Another interesting finding were intermediate resistance and susceptibility values to augmentin, aztreonan and ceftriaxone. There were 12 antimicrobial resistance patterns of which 10 were multi-resistant.

  11. Sensitivity of antibiotic resistant and antibiotic susceptible Escherichia coli, Enterococcus and Staphylococcus strains against ozone.

    Science.gov (United States)

    Heß, Stefanie; Gallert, Claudia

    2015-12-01

    Tolerance of antibiotic susceptible and antibiotic resistant Escherichia coli, Enterococcus and Staphylococcus strains from clinical and wastewater samples against ozone was tested to investigate if ozone, a strong oxidant applied for advanced wastewater treatment, will affect the release of antibiotic resistant bacteria into the aquatic environment. For this purpose, the resistance pattern against antibiotics of the mentioned isolates and their survival after exposure to 4 mg/L ozone was determined. Antibiotic resistance (AR) of the isolates was not correlating with higher tolerance against ozone. Except for ampicillin resistant E. coli strains, which showed a trend towards increased resistance, E. coli strains that were also resistant against cotrimoxazol, ciprofloxacin or a combination of the three antibiotics were similarly or less resistant against ozone than antibiotic sensitive strains. Pigment-producing Enterococcus casseliflavus and Staphylococcus aureus seemed to be more resistant against ozone than non-pigmented species of these genera. Furthermore, aggregation or biofilm formation apparently protected bacteria in subsurface layers from inactivation by ozone. The relatively large variance of tolerance against ozone may indicate that resistance to ozone inactivation most probably depends on several factors, where AR, if at all, does not play a major role.

  12. Colistin Resistance in Escherichia coli and Salmonella enterica Strains Isolated from Swine in Brazil

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    Adriano Savoia Morales

    2012-01-01

    Full Text Available Reports about acquired resistance to colistin in different bacteria species are increasing, including E. coli of animal origin, but reports of resistance in wild S. enterica of different serotypes from swine are not found in the literature. Results obtained with one hundred and twenty-six E. coli strains from diseased swine and one hundred and twenty-four S. enterica strains from diseased and carrier swine showed a frequency of 6.3% and 21% of colistin-resistant strains, respectively. When comparing the disk diffusion test with the agar dilution test to evaluate the strains, it was confirmed that the disk diffusion test is not recommended to evaluate colistin resistance as described previously. The colistin MIC 90 and MIC 50 values obtained to E. coli were 0.25 μg/mL and 0.5 μg/mL, the MIC 90 and MIC 50 to S. enterica were 1 μg/mL and 8 μg/mL. Considering the importance of colistin in control of nosocomial human infections with Gram-negative multiresistant bacteria, and the large use of this drug in animal production, the colistin resistance prevalence in enterobacteriaceae of animal origin must be monitored more closely.

  13. Comparative Study of Various E. coli Strains for Biohydrogen Production Applying Response Surface Methodology

    Directory of Open Access Journals (Sweden)

    Péter Bakonyi

    2012-01-01

    Full Text Available The proper strategy to establish efficient hydrogen-producing biosystems is the biochemical, physiological characterization of hydrogen-producing microbes followed by metabolic engineering in order to give extraordinary properties to the strains and, finally, bioprocess optimization to realize enhanced hydrogen fermentation capability. In present paper, it was aimed to show the utility both of strain engineering and process optimization through a comparative study of wild-type and genetically modified E. coli strains, where the effect of two major operational factors (substrate concentration and pH on bioH2 production was investigated by experimental design and response surface methodology (RSM was used to determine the suitable conditions in order to obtain maximum yields. The results revealed that by employing the genetically engineered E. coli (DJT 135 strain under optimized conditions (pH: 6.5; Formate conc.: 1.25 g/L, 0.63 mol H2/mol formate could be attained, which was 1.5 times higher compared to the wild-type E. coli (XL1-BLUE that produced 0.42 mol H2/mol formate (pH: 6.4; Formate conc.: 1.3 g/L.

  14. Gene doctoring: a method for recombineering in laboratory and pathogenic Escherichia coli strains

    Directory of Open Access Journals (Sweden)

    Penn Charles W

    2009-12-01

    Full Text Available Abstract Background Homologous recombination mediated by the λ-Red genes is a common method for making chromosomal modifications in Escherichia coli. Several protocols have been developed that differ in the mechanisms by which DNA, carrying regions homologous to the chromosome, are delivered into the cell. A common technique is to electroporate linear DNA fragments into cells. Alternatively, DNA fragments are generated in vivo by digestion of a donor plasmid with a nuclease that does not cleave the host genome. In both cases the λ-Red gene products recombine homologous regions carried on the linear DNA fragments with the chromosome. We have successfully used both techniques to generate chromosomal mutations in E. coli K-12 strains. However, we have had limited success with these λ-Red based recombination techniques in pathogenic E. coli strains, which has led us to develop an enhanced protocol for recombineering in such strains. Results Our goal was to develop a high-throughput recombineering system, primarily for the coupling of genes to epitope tags, which could also be used for deletion of genes in both pathogenic and K-12 E. coli strains. To that end we have designed a series of donor plasmids for use with the λ-Red recombination system, which when cleaved in vivo by the I-SceI meganuclease generate a discrete linear DNA fragment, allowing for C-terminal tagging of chromosomal genes with a 6 × His, 3 × FLAG, 4 × ProteinA or GFP tag or for the deletion of chromosomal regions. We have enhanced existing protocols and technologies by inclusion of a cassette conferring kanamycin resistance and, crucially, by including the sacB gene on the donor plasmid, so that all but true recombinants are counter-selected on kanamycin and sucrose containing media, thus eliminating the need for extensive screening. This method has the added advantage of limiting the exposure of cells to the potential damaging effects of the λ-Red system, which can lead

  15. Escherichia coli Probiotic Strain ED1a in Pigs Has a Limited Impact on the Gut Carriage of Extended-Spectrum-β-Lactamase-Producing E. coli.

    Science.gov (United States)

    Mourand, G; Paboeuf, F; Fleury, M A; Jouy, E; Bougeard, S; Denamur, E; Kempf, I

    2017-01-01

    Four trials were conducted to evaluate the impact of Escherichia coli probiotic strain ED1a administration to pigs on the gut carriage or survival in manure of extended-spectrum-β-lactamase-producing E. coli Groups of pigs were orally inoculated with strain E. coli M63 carrying the blaCTX-M-1 gene (n = 84) or used as a control (n = 26). In the first two trials, 24 of 40 E. coli M63-inoculated pigs were given E. coli ED1a orally for 6 days starting 8 days after oral inoculation. In the third trial, 10 E. coli M63-inoculated pigs were given either E. coli ED1a or probiotic E. coli Nissle 1917 for 5 days. In the fourth trial, E. coli ED1a was given to a sow and its 12 piglets, and these 12 piglets plus 12 piglets that had not received E. coli ED1a were then inoculated with E. coli M63. Fecal shedding of cefotaxime-resistant Enterobacteriaceae (CTX-RE) was studied by culture, and blaCTX-M-1 genes were quantified by PCR. The persistence of CTX-RE in manure samples from inoculated pigs or manure samples inoculated in vitro with E. coli M63 with or without probiotics was studied. The results showed that E. coli M63 and ED1a were good gut colonizers. The reduction in the level of fecal excretion of CTX-RE in E. coli ED1a-treated pigs compared to that in nontreated pigs was usually less than 1 log10 CFU and was mainly observed during the probiotic administration period. The results obtained with E. coli Nissle 1917 did not differ significantly from those obtained with E. coli ED1a. CTX-RE survival did not differ significantly in manure samples with or without probiotic treatment. In conclusion, under our experimental conditions, E. coli ED1a and E. coli Nissle 1917 could not durably prevent CTX-RE colonization of the pig gut. Copyright © 2016 American Society for Microbiology.

  16. Diffusely adherent Escherichia coli strains isolated from children and adults constitute two different populations

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    Mansan-Almeida Rosane

    2013-02-01

    Full Text Available Abstract Background Diffusely adherent Escherichia coli (DAEC have been considered a diarrheagenic category of E. coli for which several potential virulence factors have been described in the last few years. Despite this, epidemiological studies involving DAEC have shown inconsistent results. In this work, two different collections of DAEC possessing Afa/Dr genes, from children and adults, were studied regarding characteristics potentially associated to virulence. Results DAEC strains were recovered in similar frequencies from diarrheic and asymptomatic children, and more frequently from adults with diarrhea (P Citrobacter freundii strain have shown an improved ability to form biofilms in relation to the monocultures. Control strains have shown a greater diversity of Afa/Dr adhesins and higher frequencies of cellulose, TTSS, biofilm formation and induction of IL-8 secretion than strains from cases of diarrhea in children. Conclusions DAEC strains possessing Afa/Dr genes isolated from children and adults represent two different bacterial populations. DAEC strains carrying genes associated to virulence can be found as part of the normal microbiota present in asymptomatic children.

  17. Phenotypical characterization and adhesin identification in Escherichia coli strains isolated from dogs with urinary tract infections

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    Renato Pariz Maluta

    2012-03-01

    Full Text Available Pathogenic strains of Escherichia coli are the most common bacteria associated with urinary tract infections in both humans and companion animals. Standard biochemical tests may be useful in demonstrating detailed phenotypical characteristics of these strains. Thirteen strains of E. coli isolated from dogs with UTIs were submitted to biochemical tests, serotyping for O and H antigens and antimicrobial resistance testing. Furthermore, the presence of papC, sfa, and afa genes was evaluated by PCR, and genetic relationships were established using enterobacterial repetitive intergenic consensus PCR (ERIC-PCR. The antimicrobial that showed the highest resistance rate among the isolates was nalidixic acid (76.9%, followed by cephalotin (69.2%, sulfamethoxazole + trimethoprim (61.5%, tetracycline (61.5%, streptomycin (53.8%, ciprofloxacin (53.8%, ampicillin (46.2%, gentamicin (30.8% and chloramphenicol (23.1%. No isolate was resistant either to meropenem or nitrofurantoin. Among the five clusters that were identified using ERIC-PCR, one cluster (A had only one strain, which belonged to a serotype with zoonotic potential (O6:H31 and showed the genes papC+, sfa+, afa-. Strains with the genes papC-, sfa+, afa- were found in two other clusters (C and D, whereas all strains in clusters B and E possessed papC-, sfa-, afa- genes. Sucrose and raffinose phenotypic tests showed some ability in discriminating clusters A, B and C from clusters D and E.

  18. Development of glycerol-utilizing Escherichia coli strain for the production of bioethanol.

    Science.gov (United States)

    Thapa, Laxmi Prasad; Lee, Sang Jun; Yoo, Hah Young; Choi, Han Suk; Park, Chulhwan; Kim, Seung Wook

    2013-08-15

    The production of bioethanol was studied using recombinant Escherichia coli with glycerol as a carbon source. Glycerol is an attractive feedstock for biofuels production since it is generated as a major byproduct in biodiesel industry; therefore, we investigated the conversion of glycerol to bioethanol using E. coli BL21 (DE3) which harbors several genes in ethanol production pathway of Enterobacter aerogenes KCTC 2190. Fermentation was carried out at 34°C for 42h, pH 7.6, using defined production medium. Under optimal conditions, bioethanol production by the recombinant E. coli BL21 (DE3), strain pEB, was two-fold (3.01g/L) greater than that (1.45g/L) by the wild-type counterpart. The results obtained in this study will provide valuable guidelines for engineering bioethanol producers.

  19. Host Response to Porcine Strains of Escherichia coli in a Novel Pyelonephritis Model

    DEFF Research Database (Denmark)

    Isling, L. K.; Aalbæk, B.; Birck, M. M.;

    2011-01-01

    . coli strain LK67 (P fimbriae PapGI), LK76 (type 1 fimbriae) or LK82 (type 1 fimbriae and P fimbriae PapGII/III), respectively. The contralateral kidneys were inoculated with saline and served as controls. Pigs were killed 6h post-inoculation (hpi). Differential leucocyte counts, serum biochemical...... kidneys. Gross and microscopical lesions of acute pyelonephritis were demonstrated in all but one kidney inoculated with E. coli, but in none of the control kidneys. Renal parenchymal infiltration with both neutrophils and mononuclear cells, primarily CD3+ T lymphocytes, was observed at 6hpi. Most T...... lymphocytes were CD8+. Pigs in group C had the highest mean pathology scores. Neutrophils were the dominant renal leucocyte in this group, while the number of mononuclear cells was at least equal to the number of neutrophils in the lesions of pigs from groups A and B. Kidneys with a high number of E. coli had...

  20. Multirresistência antimicrobiana em cepas de Escherichia coli isoladas de cadelas com piometra Antimicrobial multi-resistance of Escherichia coli strains isolated from bitches with pyometra

    Directory of Open Access Journals (Sweden)

    V.M. Lara

    2008-08-01

    Full Text Available The antimicrobial sensibility of Escherichia coli strains isolated from the uterine content of bitches was evaluated. Fifteen E. coli strains were tested in relation to their susceptibility to different antimicrobials. The results demonstrated 100% of resistance to all tested drugs, being a quite conflicting finding compared to other works, which observed variable resistance of those bacteria to different antimicrobials but not the same multi-resistance pattern. The detection of those multi-resistance strains configures a problem, with important implications on the antimicrobial therapy. Therefore, additional investigations for a best characterization and extension of this problem are needed.

  1. ANTIMICROBIAL DRUG RESISTANCE IN STRAINS OF Escherichia coli ISOLATED FROM FOOD SOURCES

    Directory of Open Access Journals (Sweden)

    Mohammed Uddin Rasheed

    2014-07-01

    Full Text Available A variety of foods and environmental sources harbor bacteria that are resistant to one or more antimicrobial drugs used in medicine and agriculture. Antibiotic resistance in Escherichia coli is of particular concern because it is the most common Gram-negative pathogen in humans. Hence this study was conducted to determine the antibiotic sensitivity pattern of E. coli isolated from different types of food items collected randomly from twelve localities of Hyderabad, India. A total of 150 samples comprising; vegetable salad, raw egg-surface, raw chicken, unpasteurized milk, and raw meat were processed microbiologically to isolate E. coli and to study their antibiotic susceptibility pattern by the Kirby-Bauer method. The highest percentages of drug resistance in isolates of E. coli were detected from raw chicken (23.3% followed by vegetable salad (20%, raw meat (13.3%, raw egg-surface (10% and unpasteurized milk (6.7%. The overall incidence of drug resistant E. coli was 14.7%. A total of six (4% Extended Spectrum β-Lactamase (ESBL producers were detected, two each from vegetable salads and raw chicken, and one each from raw egg-surface and raw meat. Multidrug resistant strains of E. coli are a matter of concern as resistance genes are easily transferable to other strains. Pathogen cycling through food is very common and might pose a potential health risk to the consumer. Therefore, in order to avoid this, good hygienic practices are necessary in the abattoirs to prevent contamination of cattle and poultry products with intestinal content as well as forbidding the use of untreated sewage in irrigating vegetables.

  2. Enhancement of xylose utilization from corn stover by a recombinant Escherichia coli strain for ethanol production.

    Science.gov (United States)

    Saha, Badal C; Qureshi, Nasib; Kennedy, Gregory J; Cotta, Michael A

    2015-08-01

    Effects of substrate-selective inoculum prepared by growing on glucose, xylose, arabinose, GXA (glucose, xylose, arabinose, 1:1:1) and corn stover hydrolyzate (dilute acid pretreated and enzymatically hydrolyzed, CSH) on ethanol production from CSH by a mixed sugar utilizing recombinant Escherichia coli (strain FBR5) were investigated. The initial ethanol productivity was faster for the seed grown on xylose followed by GXA, CSH, glucose and arabinose. Arabinose grown seed took the longest time to complete the fermentation. Delayed saccharifying enzyme addition in simultaneous saccharification and fermentation of dilute acid pretreated CS by the recombinant E. coli strain FBR5 allowed the fermentation to finish in a shorter time than adding the enzyme simultaneously with xylose grown inoculum. Use of substrate selective inoculum and fermenting pentose sugars first under glucose limited condition helped to alleviate the catabolite repression of the recombinant bacterium on ethanol production from lignocellulosic hydrolyzate.

  3. Implications of alpha-amylase production and beta-glucuronidase expression in Escherichia coli strains.

    Science.gov (United States)

    Caldini, G; Strappini, C; Trotta, F; Cenci, G

    1999-01-01

    Two Escherichia coli strains in which alpha-amylase production differed were used to study in depth some characteristics related to beta-glucuronidase induction by starch. The beta-glucuronidase background activity in Luria broth medium was comparable for the two isolates, but only amylase positive S1 was able to grow on starch molecules supplied as the sole carbon source. In this case growth resulted at higher beta-glucuronidase levels (p beta-glucuronidase activity of amylase negative M94 remained unchanged during starvation on starch medium, but an induced response was observed with methylumbelliferyl-glucuronide. These results further support the hypothesis that starch metabolism is involved in the complex beta-glucuronidase regulation of E. coli strains. This is relevant not only for basic research but also to investigating gut microbial enzymology.

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

    Science.gov (United States)

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

    2013-01-01

    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 β-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 (PUPEC 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 (PUPEC 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.

  5. Modeling the microbial growth of two Escherichia coli strains in a multi-substrate environment

    OpenAIRE

    Poccia,M. E.; Beccaria, A. J.; R. G. Dondo

    2014-01-01

    The microbial growth in multi-substrate environments may be viewed as an optimal resources allocation problem. The optimization aims at maximizing some biological objective like the biomass growth. The models developed using this hypothesis are called “cybernetic” and they represent the complex cell structure as an optimizing function that regulates the intracellular enzymatic machinery. In this work, a cybernetic model was developed to represent the growth of two E. coli strains (JM 109 and ...

  6. Colistin Resistance mcr-1-Gene-Bearing Escherichia coli Strain from the United States.

    Science.gov (United States)

    Meinersmann, Richard J; Ladely, Scott R; Plumblee, Jodie R; Hall, M Carolina; Simpson, Sheron A; Ballard, Linda L; Scheffler, Brian E; Genzlinger, Linda L; Cook, Kimberly L

    2016-09-01

    Transmissible colistin resistance in the form of an mcr-1-gene-bearing plasmid has been recently reported in Enterobacteriaceae in several parts of the world. We report here the completed genome sequence of an Escherichia coli strain isolated from swine in the United States that carried the mcr-1 gene on an IncI2-type plasmid. Copyright © 2016 Meinersmann et al.

  7. Essential validation methods for E. coli strains created by chromosome engineering

    OpenAIRE

    Krishnan, S.T.; Moolman, M.C.; van Laar, T.; Meyer, A. S; Dekker, N.H.

    2015-01-01

    Background Chromosome engineering encompasses a collection of homologous recombination-based techniques that are employed to modify the genome of a model organism in a controlled fashion. Such techniques are widely used in both fundamental and industrial research to introduce multiple insertions in the same Escherichia coli strain. To date, λ-Red recombination (also known as recombineering) and P1 phage transduction are the most successfully implemented chromosome engineering techniques in E....

  8. [Purification of recombinant Bacillus cereus ResD-ResE proteins expressed in Escherichia coli strains].

    Science.gov (United States)

    Shapyrina, E V; Shadrin, A M; Solonin, A S

    2013-01-01

    Recombinant E. coli strains expressing the Bacillus cereus ATCC 14579T resD and resEgenes fused with the ubiquitin gene were constructed, and purification of the ResD and ResE proteins was performed. The approach used in the study allowed us to increase the protein yield of the electrophoretic homogeneous ResD andResE proteins without denaturation steps up to 150 mg per gram of wet cell weight.

  9. Posttranslationally caused bioluminescence burst of the Escherichia coli luciferase reporter strain.

    Science.gov (United States)

    Ideguchi, Yamato; Oshikoshi, Yuta; Ryo, Masashi; Motoki, Shogo; Kuwano, Takashi; Tezuka, Takafumi; Aoki, Setsuyuki

    2016-01-01

    We continuously monitored bioluminescence from a wild-type reporter strain of Escherichia coli (lacp::luc+/WT), which carries the promoter of the lac operon (lacp) fused with the firefly luciferase gene (luc+). This strain showed a bioluminescence burst when shifted into the stationary growth phase. Bioluminescence profiles of other wild-type reporter strains (rpsPp::luc+ and argAp::luc+) and gene-deletion reporter strains (lacp::luc+/crp- and lacp::luc+/lacI-) indicate that transcriptional regulation is not responsible for generation of the burst. Consistently, changes in the luciferase protein levels did not recapitulate the profile of the burst. On the other hand, dissolved oxygen levels increased over the period across the burst, suggesting that the burst is, at least partially, caused by an increase in intracellular oxygen levels. We discuss limits of the firefly luciferase when used as a reporter for gene expression and its potential utility for monitoring metabolic changes in cells.

  10. Environmental and genetic factors affecting mutability to aminoglycoside antibiotics among Escherichia coli K12 strains

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    Monteiro A.C.M.

    2003-01-01

    Full Text Available Environmental and genetic factors affecting the in vitro spontaneous mutation frequencies to aminoglycoside resistance in Escherichia coli K12 were investigated. Spontaneous mutation frequencies to kanamycin resistance were at least 100 fold higher on modified Luria agar (L2 plates, when compared to results obtained in experiments carried out with Nutrient agar (NA plates. In contrast to rifampincin, the increased mutability to kanamycin resistance could not be attributed to a mutator phenotype expressed by DNA repair defective strains. Kanamycin mutant selection windows and mutant preventive concentrations on L2 plates were at least fourfold higher than on NA plates, further demonstrating the role of growth medium composition on the mutability to aminoglycosides. Mutability to kanamycin resistance was increased following addition of sorbitol, suggesting that osmolarity is involved on the spontaneous mutability of E. coli K12 strains to aminoglycosides. The spontaneous mutation rates to kanamycin resistance on both L2 and NA plates were strictly associated with the selective antibiotic concentrations. Moreover, mutants selected at different antibiotic concentrations expressed heterogeneous resistance levels to kanamycin and most of them expressing multiple resistance to all tested aminoglycoside antibiotics (gentamicin, neomycin, amykacin and tobramycin. These results will contribute to a better understanding of the complex nature of aminoglycoside resistance and the emergence of spontaneous resistant mutants among E. coli K12 strains.

  11. A conserved virulence plasmidic region contributes to the virulence of the multiresistant Escherichia coli meningitis strain S286 belonging to phylogenetic group C.

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    Chloé Lemaître

    Full Text Available Recent isolation of the non-K1 Escherichia coli neonatal meningitis strain S286, belonging to phylogroup C, which is closely related to major group B1, and producing an extended-spectrum beta-lactamase, encouraged us to seek the genetic determinants responsible for its virulence. We show that S286 belongs to the sequence O type ST23O78 and harbors 4 large plasmids. The largest one, pS286colV (~120 kb, not related to resistance, contains genes characteristic of a Conserved Virulence Plasmidic (CVP region initially identified in B2 extra-intestinal avian pathogenic E. coli (APEC strains and in the B2 neonatal meningitis E. coli strain S88. The sequence of this CVP region has a strong homology (98% with that of the recently sequenced plasmid pChi7122-1 of the O78 APEC strain Chi7122. A CVP plasmid-cured variant of S286 was less virulent than the wild type strain in a neonatal rat sepsis model with a significant lower level of bacteremia at 24 h (4.1 ± 1.41 versus 2.60 ± 0.16 log CFU/ml, p = 0.001 and mortality. However, the mortality in the model of adult mice was comparable between wild type and variant indicating that pS286colV is not sufficient by itself to fully explain the virulence of S286. Gene expression analysis of pS286colV in iron depleted environment was very close to that of pS88, suggesting that genes of CVP region may be expressed similarly in two very different genetic backgrounds (group C versus group B2. Screening a collection of 178 human A/B1 extraintestinal pathogenic E. coli (ExPEC strains revealed that the CVP region is highly prevalent (23% and MLST analysis indicated that these CVP positive strains belong to several clusters and mostly to phylogroup C. The virulence of S286 is explained in part by the presence of CVP region and this region has spread in different clusters of human A/B1 ExPEC, especially in group C.

  12. A conserved virulence plasmidic region contributes to the virulence of the multiresistant Escherichia coli meningitis strain S286 belonging to phylogenetic group C.

    Science.gov (United States)

    Lemaître, Chloé; Mahjoub-Messai, Farah; Dupont, Damien; Caro, Valérie; Diancourt, Laure; Bingen, Edouard; Bidet, Philippe; Bonacorsi, Stéphane

    2013-01-01

    Recent isolation of the non-K1 Escherichia coli neonatal meningitis strain S286, belonging to phylogroup C, which is closely related to major group B1, and producing an extended-spectrum beta-lactamase, encouraged us to seek the genetic determinants responsible for its virulence. We show that S286 belongs to the sequence O type ST23O78 and harbors 4 large plasmids. The largest one, pS286colV (~120 kb), not related to resistance, contains genes characteristic of a Conserved Virulence Plasmidic (CVP) region initially identified in B2 extra-intestinal avian pathogenic E. coli (APEC) strains and in the B2 neonatal meningitis E. coli strain S88. The sequence of this CVP region has a strong homology (98%) with that of the recently sequenced plasmid pChi7122-1 of the O78 APEC strain Chi7122. A CVP plasmid-cured variant of S286 was less virulent than the wild type strain in a neonatal rat sepsis model with a significant lower level of bacteremia at 24 h (4.1 ± 1.41 versus 2.60 ± 0.16 log CFU/ml, p = 0.001) and mortality. However, the mortality in the model of adult mice was comparable between wild type and variant indicating that pS286colV is not sufficient by itself to fully explain the virulence of S286. Gene expression analysis of pS286colV in iron depleted environment was very close to that of pS88, suggesting that genes of CVP region may be expressed similarly in two very different genetic backgrounds (group C versus group B2). Screening a collection of 178 human A/B1 extraintestinal pathogenic E. coli (ExPEC) strains revealed that the CVP region is highly prevalent (23%) and MLST analysis indicated that these CVP positive strains belong to several clusters and mostly to phylogroup C. The virulence of S286 is explained in part by the presence of CVP region and this region has spread in different clusters of human A/B1 ExPEC, especially in group C.

  13. Enterohaemolysin production and verotoxin genes in Esche-richia coli strains isolated from neonatal calves in India

    Institute of Scientific and Technical Information of China (English)

    Diganta Pan; Ashok Kumar Bhatia; KNBhilegaonkar

    2009-01-01

    Objective:To screened all haemolytic strains of Escherichia coli (E.coli)for the presence of verotoxin genes and speculate the association between enterohaemolysin production and presence of verotoxin genes.Methods:A total 176 of E.coli strains of 64 serogroups isolated from neonatal calves were selected and screened for al-pha-haemolysin and enterohaemolysin production on sheep blood agar and 5% washed sheep blood agar.Two types of haemolytic strains were further characterized by PCR for presence of Verotoxin gene (VT1 and VT2) and confirmed by verocell cytotoxicity assay.Results:Among 27 enterohaemolytic positive strains,19 (70. 37%)strains were found positive for presence of verotoxins (VTs)gene and verocell cytotoxicity assay. Whereas 34 alpha-haemolysin strains were found negative for VTs.Eight strains of E.coli were found VTs neg-ative but Ehx positive.Conclusion:The close association between enterohaemolysin production and presence of verotoxin genes makes it useful epidemiological marker for rapid screening of Verotoxic E.coli but this pheno-typic marker for screening of verotoxigenic Escherichia coli(VTEC)alone may raise a question in animal.

  14. Synthesis and accumulation of aromatic aldehydes in an engineered strain of Escherichia coli.

    Science.gov (United States)

    Kunjapur, Aditya M; Tarasova, Yekaterina; Prather, Kristala L J

    2014-08-20

    Aromatic aldehydes are useful in numerous applications, especially as flavors, fragrances, and pharmaceutical precursors. However, microbial synthesis of aldehydes is hindered by rapid, endogenous, and redundant conversion of aldehydes to their corresponding alcohols. We report the construction of an Escherichia coli K-12 MG1655 strain with reduced aromatic aldehyde reduction (RARE) that serves as a platform for aromatic aldehyde biosynthesis. Six genes with reported activity on the model substrate benzaldehyde were rationally targeted for deletion: three genes that encode aldo-keto reductases and three genes that encode alcohol dehydrogenases. Upon expression of a recombinant carboxylic acid reductase in the RARE strain and addition of benzoate during growth, benzaldehyde remained in the culture after 24 h, with less than 12% conversion of benzaldehyde to benzyl alcohol. Although individual overexpression results demonstrated that all six genes could contribute to benzaldehyde reduction in vivo, additional experiments featuring subset deletion strains revealed that two of the gene deletions were dispensable under the conditions tested. The engineered strain was next investigated for the production of vanillin from vanillate and succeeded in preventing formation of the byproduct vanillyl alcohol. A pathway for the biosynthesis of vanillin directly from glucose was introduced and resulted in a 55-fold improvement in vanillin titer when using the RARE strain versus the wild-type strain. Finally, synthesis of the chiral pharmaceutical intermediate L-phenylacetylcarbinol (L-PAC) was demonstrated from benzaldehyde and glucose upon expression of a recombinant mutant pyruvate decarboxylase in the RARE strain. Beyond allowing accumulation of aromatic aldehydes as end products in E. coli, the RARE strain expands the classes of chemicals that can be produced microbially via aldehyde intermediates.

  15. The investigation of new forms of resistance to some antibiotics in E coli strains isolated from piglets

    Directory of Open Access Journals (Sweden)

    Ašanin Ružica

    2009-01-01

    Full Text Available The resistance of bacteria poses a significant problem everywhere in the world, and consequently in our country as well. The non-critical use of antibacterial medicines in human and veterinary medicine has contributed to the spreading of this resistance. Due to the topical importance of this problem, large numbers of states in the world are financing projects of which the objective is to follow and monitor bacterial resistance. The objective of this investigation was to isolate and identify pathogenic strains of E. coli from piglets with clinically manifest diarrhea and to examine the sensitivity of the isolated strains to a certain number of selected antibiotics. The material for these investigations were parts of intestines (jejunum, ileum from piglets that died, rectal smears and feces of diseased piglets sampled pig farms in the vicinity of Belgrade. Conventional methods of microbiological diagnostics were used for isolation, and conventional and commercial tests API 20E (bio Merieux, France were used for identification. Following biochemical identification using hyper immune serums for certain group (O antigens: (O8, O138, O139, O147, O149, and O157, the serological typization of the strains was carried out. Commercial antiserums: T K88 (F4, K99 (F5, and 987P (F6, Toxigenic E. coli pili antisera, Denka Seiken Co. Ltd. Tokyo, Japan were used to establish the presence and to identify fimbrial antigens-adhesions through slide agglutination reaction. The sensitivity of the antibiotics to the isolated strains of E. coli was examined using the disc diffusion method according to Kirby Bauer and the microdillution method in bouillon according to CLSI recommendations (2008. Examinations using the microdillution method in bouillon were performed with pure active antibiotic substances: ampicillin, apramycin, gentamicin, kanamycin, tetracycline, ceftriaxone, and ciprofloxacin (Sigma, USA. A total of 400 E. coli strains were isolated, including 48 E

  16. Biofilm formation and sanitizer resistance of Escherichia coli 0157:H7 strains isolated from "High Event Period" meat contamination

    Science.gov (United States)

    In the meat industry, a “High Event Period” (HEP) is defined as a time period during which commercial meat plants experience a higher than usual rate of E. coli O157:H7 contamination. Genetic analysis indicated that within a HEP, most of the E. coli O157:H7 strains belong to a singular dominant str...

  17. Global gene expression profiling of the asymptomatic bacteriuria Escherichia coli strain 83972 in the human urinary tract

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Klemm, Per

    2006-01-01

    Urinary tract infections (UTIs) are an important health problem worldwide, with many million cases each year. Escherichia coli is the most common organism causing UTIs in humans. The asymptomatic bacteriuria E. coli strain 83972 is an excellent colonizer of the human urinary tract, where it causes...

  18. Sequencing and functional annotation of avian pathogenic Escherichia coli serogroup O78 strains reveal the evolution of E. coli lineages pathogenic for poultry via distinct mechanisms.

    Science.gov (United States)

    Dziva, Francis; Hauser, Heidi; Connor, Thomas R; van Diemen, Pauline M; Prescott, Graham; Langridge, Gemma C; Eckert, Sabine; Chaudhuri, Roy R; Ewers, Christa; Mellata, Melha; Mukhopadhyay, Suman; Curtiss, Roy; Dougan, Gordon; Wieler, Lothar H; Thomson, Nicholas R; Pickard, Derek J; Stevens, Mark P

    2013-03-01

    Avian pathogenic Escherichia coli (APEC) causes respiratory and systemic disease in poultry. Sequencing of a multilocus sequence type 95 (ST95) serogroup O1 strain previously indicated that APEC resembles E. coli causing extraintestinal human diseases. We sequenced the genomes of two strains of another dominant APEC lineage (ST23 serogroup O78 strains χ7122 and IMT2125) and compared them to each other and to the reannotated APEC O1 sequence. For comparison, we also sequenced a human enterotoxigenic E. coli (ETEC) strain of the same ST23 serogroup O78 lineage. Phylogenetic analysis indicated that the APEC O78 strains were more closely related to human ST23 ETEC than to APEC O1, indicating that separation of pathotypes on the basis of their extraintestinal or diarrheagenic nature is not supported by their phylogeny. The accessory genome of APEC ST23 strains exhibited limited conservation of APEC O1 genomic islands and a distinct repertoire of virulence-associated loci. In light of this diversity, we surveyed the phenotype of 2,185 signature-tagged transposon mutants of χ7122 following intra-air sac inoculation of turkeys. This procedure identified novel APEC ST23 genes that play strain- and tissue-specific roles during infection. For example, genes mediating group 4 capsule synthesis were required for the virulence of χ7122 and were conserved in IMT2125 but absent from APEC O1. Our data reveal the genetic diversity of E. coli strains adapted to cause the same avian disease and indicate that the core genome of the ST23 lineage serves as a chassis for the evolution of E. coli strains adapted to cause avian or human disease via acquisition of distinct virulence genes.

  19. Biofilm formation and sanitizer resistance of Escherichia coli O157:H7 strains isolated from "high event period" meat contamination.

    Science.gov (United States)

    Wang, Rong; Kalchayanand, Norasak; King, David A; Luedtke, Brandon E; Bosilevac, Joseph M; Arthur, Terrance M

    2014-11-01

    In the meat industry, a "high event period" (HEP) is defined as a time period during which commercial meat plants experience a higher than usual rate of Escherichia coli O157:H7 contamination. Genetic analysis indicated that within a HEP, most of the E. coli O157:H7 strains belong to a singular dominant strain type. This was in disagreement with the current beef contamination model stating that contamination occurs when incoming pathogen load on animal hides, which consists of diverse strain types of E. coli O157:H7, exceeds the intervention capacity. Thus, we hypothesize that the HEP contamination may be due to certain in-plant colonized E. coli O157:H7 strains that are better able to survive sanitization through biofilm formation. To test our hypothesis, a collection of 45 E. coli O157:H7 strains isolated from HEP beef contamination incidents and a panel of 47 E. coli O157:H7 strains of diverse genetic backgrounds were compared for biofilm formation and sanitizer resistance. Biofilm formation was tested on 96-well polystyrene plates for 1 to 6 days. Biofilm cell survival and recovery growth after sanitization were compared between the two strain collections using common sanitizers, including quaternary ammonium chloride, chlorine, and sodium chlorite. No difference in "early stage" biofilms was observed between the two strain collections after incubation at 22 to 25°C for 1 or 2 days. However, the HEP strains demonstrated significantly higher potency of "mature" biofilm formation after incubation for 4 to 6 days. Biofilms of the HEP strains also exhibited significantly stronger resistance to sanitization. These data suggest that biofilm formation and sanitization resistance could have a role in HEP beef contamination by E. coli O157:H7, which highlights the importance of proper and complete sanitization of food contact surfaces and food processing equipment in commercial meat plants.

  20. Behavior of shiga toxin-producing Escherichia coli, enteroinvasive E. coli, enteropathogenic E. coli and enterotoxigenic E. coli strains on whole and sliced jalapeño and serrano peppers.

    Science.gov (United States)

    Gómez-Aldapa, Carlos A; Rangel-Vargas, Esmeralda; Gordillo-Martínez, Alberto J; Castro-Rosas, Javier

    2014-06-01

    The behavior of enterotoxigenic Escherichia coli (ETEC), enteropathogenic E. coli (EPEC), enteroinvasive E. coli (EIEC) and non-O157 shiga toxin-producing E. coli (non-O157-STEC) on whole and slices of jalapeño and serrano peppers as well as in blended sauce at 25 ± 2 °C and 3 ± 2 °C was investigated. Chili peppers were collected from markets of Pachuca city, Hidalgo, Mexico. On whole serrano and jalapeño stored at 25 ± 2 °C or 3 ± 2 °C, no growth was observed for EPEC, ETEC, EIEC and non-O157-STEC rifampicin resistant strains. After twelve days at 25 ± 2 °C, on serrano peppers all diarrheagenic E. coli pathotypes (DEP) strains had decreased by a total of approximately 3.7 log, whereas on jalapeño peppers the strains had decreased by approximately 2.8 log, and at 3 ± 2 °C they decreased to approximately 2.5 and 2.2 log respectively, on serrano and jalapeño. All E. coli pathotypes grew onto sliced chili peppers and in blended sauce: after 24 h at 25 ± 2 °C, all pathotypes had grown to approximately 3 and 4 log CFU on pepper slices and sauce, respectively. At 3 ± 2 °C the bacterial growth was inhibited. Copyright © 2014 Elsevier Ltd. All rights reserved.

  1. A virulent parent with probiotic progeny: comparative genomics of Escherichia coli strains CFT073, Nissle 1917 and ABU 83972

    DEFF Research Database (Denmark)

    Vejborg, Rebecca Munk; Friis, Carsten; Hancock, Viktoria

    2010-01-01

    Escherichia coli is a highly versatile species encompassing a diverse spectrum of strains, i.e. from highly virulent isolates causing serious infectious diseases to commensals and probiotic strains. Although much is known about bacterial pathogenicity in E. coli, the understanding of which genetic......) with the highly virulent uropathogen CFT073. Only relatively minor genetic variations were found between the isolates, suggesting that the three strains may have originated from the same virulent ancestral parent. Interestingly, Nissle 1917 (a gut commensal strain) was more similar to CFT073 with respect...

  2. Metabolic engineering of a reduced-genome strain of Escherichia coli for L-threonine production

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    Yoon Byoung

    2009-01-01

    Full Text Available Abstract Background Deletion of large blocks of nonessential genes that are not needed for metabolic pathways of interest can reduce the production of unwanted by-products, increase genome stability, and streamline metabolism without physiological compromise. Researchers have recently constructed a reduced-genome Escherichia coli strain MDS42 that lacks 14.3% of its chromosome. Results Here we describe the reengineering of the MDS42 genome to increase the production of the essential amino acid L-threonine. To this end, we over-expressed a feedback-resistant threonine operon (thrA*BC, deleted the genes that encode threonine dehydrogenase (tdh and threonine transporters (tdcC and sstT, and introduced a mutant threonine exporter (rhtA23 in MDS42. The resulting strain, MDS-205, shows an ~83% increase in L-threonine production when cells are grown by flask fermentation, compared to a wild-type E. coli strain MG1655 engineered with the same threonine-specific modifications described above. And transcriptional analysis revealed the effect of the deletion of non-essential genes on the central metabolism and threonine pathways in MDS-205. Conclusion This result demonstrates that the elimination of genes unnecessary for cell growth can increase the productivity of an industrial strain, most likely by reducing the metabolic burden and improving the metabolic efficiency of cells.

  3. Polyhydroxyalkanoate production from sucrose by Cupriavidus necator strains harboring csc genes from Escherichia coli W.

    Science.gov (United States)

    Arikawa, Hisashi; Matsumoto, Keiji; Fujiki, Tetsuya

    2017-09-09

    Cupriavidus necator H16 is the most promising bacterium for industrial production of polyhydroxyalkanoates (PHAs) because of their remarkable ability to accumulate them in the cells. With genetic modifications, this bacterium can produce poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) (PHBHHx), which has better physical properties, as well as poly(3-hydroxybutyrate) (PHB) using plant oils and sugars as a carbon source. Considering production cost, sucrose is a very attractive raw material because it is inexpensive; however, this bacterium cannot assimilate sucrose. Here, we used the sucrose utilization (csc) genes of Escherichia coli W to generate C. necator strains that can assimilate sucrose. Especially, glucose-utilizing recombinant C. necator strains harboring the sucrose hydrolase gene (cscA) and sucrose permease gene (cscB) of E. coli W grew well on sucrose as a sole carbon source and accumulated PHB. In addition, strains introduced with a crotonyl-CoA reductase gene (ccr), ethylmalonyl-CoA decarboxylase gene (emd), and some other genetic modifications besides the csc genes and the glucose-utilizing mutations produced PHBHHx with a 3-hydroxyhexanoate (3HHx) content of maximum approximately 27 mol% from sucrose. Furthermore, when one of the PHBHHx-producing strains was cultured with sucrose solution in a fed-batch fermentation, PHBHHx with a 3HHx content of approximately 4 mol% was produced and reached 113 g/L for 65 h, which is approximately 1.5-fold higher than that produced using glucose solution.

  4. A genomically modified Escherichia coli strain carrying an orthogonal E. coli histidyl-tRNA synthetase•tRNA(His) pair.

    Science.gov (United States)

    Englert, Markus; Vargas-Rodriguez, Oscar; Reynolds, Noah M; Wang, Yane-Shih; Söll, Dieter; Umehara, Takuya

    2017-03-10

    Development of new aminoacyl-tRNA synthetase (aaRS)•tRNA pairs is central for incorporation of novel non-canonical amino acids (ncAAs) into proteins via genetic code expansion (GCE). The Escherichia coli and Caulobacter crescentus histidyl-tRNA synthetases (HisRS) evolved divergent mechanisms of tRNA(His) recognition that prevent their cross-reactivity. Although the E. coli HisRS•tRNA(His) pair is a good candidate for GCE, its use in C. crescentus is limited by the lack of established genetic selection methods and by the low transformation efficiency of C. crescentus. E. coli was genetically engineered to use a C. crescentus HisRS•tRNA(His) pair. Super-folder green fluorescent protein (sfGFP) and chloramphenicol acetyltransferase (CAT) were used as reporters for read-through assays. A library of 313 ncAAs coupled with the sfGFP reporter system was employed to investigate the specificity of E. coli HisRS in vivo. A genomically modified E. coli strain (named MEOV1) was created. MEVO1 requires an active C. crescentus HisRS•tRNA(His) pair for growth, and displays a similar doubling time as the parental E. coli strain. sfGFP- and CAT-based assays showed that the E. coli HisRS•tRNA(His) pair is orthogonal in MEOV1 cells. A mutation in the anticodon loop of E. coli tRNA(His)CUA elevated its suppression efficiency by 2-fold. The C. crescentus HisRS•tRNA(His) pair functionally complements an E. coli ΔhisS strain. The E. coli HisRS•tRNA(His) is orthogonal in MEOV1 cells. E. coli tRNA(His)CUA is an efficient amber suppressor in MEOV1. We developed a platform that allows protein engineering of E. coli HisRS that should facilitate GCE in E. coli. This article is part of a Special Issue entitled "Biochemistry of Synthetic Biology - Recent Developments" Guest Editor: Dr. Ilka Heinemann and Dr. Patrick O'Donoghue. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Chicken Meat as a Reservoir of Colistin-Resistant Escherichia coli Strains Carrying mcr-1 Genes in South America.

    Science.gov (United States)

    Monte, Daniel Farias; Mem, Andressa; Fernandes, Miriam R; Cerdeira, Louise; Esposito, Fernanda; Galvão, Julia A; Franco, Bernadette D G M; Lincopan, Nilton; Landgraf, Mariza

    2017-05-01

    The detection and rapid spread of colistin-resistant Enterobacteriaceae carrying the mcr-1 gene has created an urgent need to strengthen surveillance. In this study, eight clonally unrelated colistin-resistant Escherichia coli isolates carrying mcr-1 and blaCTX-M or blaCMY-2 genes were isolated from commercial chicken meat in Brazil. Most E. coli strains carried IncX4 plasmids, previously identified in human and animal isolates. These results highlight a new reservoir of mcr-1-harboring E. coli strains in South America. Copyright © 2017 American Society for Microbiology.

  6. A versatile Escherichia coli strain for identification of biotin transporters and for biotin quantification.

    Science.gov (United States)

    Finkenwirth, Friedrich; Kirsch, Franziska; Eitinger, Thomas

    2014-01-01

    Biotin is an essential cofactor of carboxylase enzymes in all kingdoms of life. The vitamin is produced by many prokaryotes, certain fungi, and plants. Animals depend on biotin uptake from their diet and in humans lack of the vitamin is associated with serious disorders. Many aspects of biotin metabolism, uptake, and intracellular transport remain to be elucidated. In order to characterize the activity of novel biotin transporters by a sensitive assay, an Escherichia coli strain lacking both biotin synthesis and its endogenous high-affinity biotin importer was constructed. This strain requires artificially high biotin concentrations for growth. When only trace levels of biotin are available, it is viable only if equipped with a heterologous high-affinity biotin transporter. This feature was used to ascribe transport activity to members of the BioY protein family in previous work. Here we show that this strain together with its parent is also useful as a diagnostic tool for wide-concentration-range bioassays.

  7. Strain dependent UV degradation of Escherichia coli DNA monitored by Fourier transform infrared spectroscopy.

    Science.gov (United States)

    Muntean, Cristina M; Lapusan, Alexandra; Mihaiu, Liora; Stefan, Razvan

    2014-01-05

    In this work we present a method for detection of DNA isolated from nonpathogenic Escherichia coli strains, respectively. Untreated and UV irradiated bacterial DNAs were analyzed by FT-IR spectroscopy, to investigate their screening characteristic features and their structural radiotolerance at 253.7nm. FT-IR spectra, providing a high molecular structural information, have been analyzed in the wavenumber range 800-1800cm(-1). FT-IR signatures, spectroscopic band assignments and structural interpretations of these DNAs are reported. Also, UV damage at the DNA molecular level is of interest. Strain dependent UV degradation of DNA from E. coli has been observed. Particularly, alterations in nucleic acid bases, base pairing and base stacking have been found. Also changes in the DNA conformation and deoxyribose were detected. Based on this work, specific E. coli DNA-ligand interactions, drug development and vaccine design for a better understanding of the infection mechanism caused by an interference between pathogenic and nonpathogenic bacteria and for a better control of disease, respectively, might be further investigated using Fourier transform infrared spectroscopy. Besides, understanding the pathways for UV damaged DNA response, like nucleic acids repair mechanisms is appreciated.

  8. Antimicrobial Susceptibility Testing for Escherichia coli Strains to Fluoroquinolones, in Urinary Tract Infections

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    FA Nakhjavani

    2007-05-01

    Full Text Available Background: Urinary Tract Infections (UTIs are one of the most common infectious diseases diagnosed all over the world. Meanwhile most episode of UTIs are caused by Escherichia coli (up to 85% and frequently fluoroquinolones are preferred as initial agents for empiric therapy of UTIs. Widespread use of fluoroquinolones has resulted in an increasing incidence of resistance these agents all over the world. The aim of this study was to assess, susceptibility of Escherichia coli strains from UTI patients against common fluoroquinolones. Methods: Antimicrobial susceptibility testing was determined by disk agar diffusion (DAD and Minimal Inhibitory Concentration methods as described by the National Committee for Clinical Laboratory Standards (NCCLS. Results: One hundred sixty four clinical isolates of E. coli were collected by urine cultures from patients with UTI. The extent of resistant to nalidixic acid, ofloxacin, norfloxacin and ciprofloxacin, by disk diffusion method was 49.3%, 44.5%, 41.4% and 40.2%, respectively. Resistance to ciprofloxacin by MIC method was 4.9%. Conclusion: This study represents high level resistant of E. coli isolates from UTI patients. It is because of inappropriate and incorrect administration of antimicrobial agents in blind cases. This problem remarks significance of performing antimicrobial susceptibility testing before empiric antibiotic therapy. To overcome this problem use of unnecessary antibiotics therapy should be limited.

  9. A regulatory trade-off as a source of strain variation in the species Escherichia coli.

    Science.gov (United States)

    King, Thea; Ishihama, Akira; Kori, Ayako; Ferenci, Thomas

    2004-09-01

    There are few existing indications that strain variation in prokaryotic gene regulation is common or has evolutionary advantage. In this study, we report on isolates of Escherichia coli with distinct ratios of sigma factors (RpoD, sigmaD, or sigma70 and RpoS or sigmaS) that affect transcription initiated by RNA polymerase. Both laboratory E. coli K-12 lineages and nondomesticated isolates exhibit strain-specific endogenous levels of RpoS protein. We demonstrate that variation in genome usage underpins intraspecific variability in transcription patterns, resistance to external stresses, and the choice of beneficial mutations under nutrient limitation. Most unexpectedly, RpoS also controlled strain variation with respect to the metabolic capability of bacteria with more than a dozen carbon sources. Strains with higher sigmaS levels were more resistant to external stress but metabolized fewer substrates and poorly competed for low concentrations of nutrients. On the other hand, strains with lower sigmaS levels had broader nutritional capabilities and better competitive ability with low nutrient concentrations but low resistance to external stress. In other words, RpoS influenced both r and K strategist functions of bacteria simultaneously. The evolutionary principle driving strain variation is proposed to be a conceptually novel trade-off that we term SPANC (for "self-preservation and nutritional competence"). The availability of multiple SPANC settings potentially broadens the niche occupied by a species consisting of individuals with narrow specialization and reveals an evolutionary advantage offered by polymorphic regulation. Regulatory diversity is likely to be a significant contributor to complexity in a bacterial world in which multiple sigma factors are a universal feature.

  10. Impact of individual mutations on increased fitness in adaptively evolved strains of Escherichia coli.

    Science.gov (United States)

    Applebee, M Kenyon; Herrgård, Markus J; Palsson, Bernhard Ø

    2008-07-01

    We measured the relative fitness among a set of experimentally evolved Escherichia coli strains differing by a small number of adaptive mutations by directly measuring allelic frequencies in head-to-head competitions using a mass spectrometry-based method. We compared the relative effects of mutations to the same or similar genes acquired in multiple strains when expressed in allele replacement strains. We found that the strongest determinant of fitness among the evolved strains was the impact of beneficial mutations to the RNA polymerase beta and beta' subunit genes. We also identified several examples of epistatic interactions between rpoB/C and glpK mutations and identified two other mutations that are beneficial only in the presence of previously acquired mutations but that have little or no adaptive benefit to the wild-type strain. Allele frequency estimation is shown to be a highly sensitive method for measuring selection rates during competitions between strains differing by as little as a single-nucleotide polymorphism and may be of great use for investigating epistatic interactions.

  11. Infections with avian pathogenic and fecal Escherichia coli strains display similar lung histopathology and macrophage apoptosis.

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    Fabiana Horn

    Full Text Available The purpose of this study was to compare histopathological changes in the lungs of chickens infected with avian pathogenic (APEC and avian fecal (A(fecal Escherichia coli strains, and to analyze how the interaction of the bacteria with avian macrophages relates to the outcome of the infection. Chickens were infected intratracheally with three APEC strains, MT78, IMT5155, and UEL17, and one non-pathogenic A(fecal strain, IMT5104. The pathogenicity of the strains was assessed by isolating bacteria from lungs, kidneys, and spleens at 24 h post-infection (p.i.. Lungs were examined for histopathological changes at 12, 18, and 24 h p.i. Serial lung sections were stained with hematoxylin and eosin (HE, terminal deoxynucleotidyl dUTP nick end labeling (TUNEL for detection of apoptotic cells, and an anti-O2 antibody for detection of MT78 and IMT5155. UEL17 and IMT5104 did not cause systemic infections and the extents of lung colonization were two orders of magnitude lower than for the septicemic strains MT78 and IMT5155, yet all four strains caused the same extent of inflammation in the lungs. The inflammation was localized; there were some congested areas next to unaffected areas. Only the inflamed regions became labeled with anti-O2 antibody. TUNEL labeling revealed the presence of apoptotic cells at 12 h p.i in the inflamed regions only, and before any necrotic foci could be seen. The TUNEL-positive cells were very likely dying heterophils, as evidenced by the purulent inflammation. Some of the dying cells observed in avian lungs in situ may also be macrophages, since all four avian E. coli induced caspase 3/7 activation in monolayers of HD11 avian macrophages. In summary, both pathogenic and non-pathogenic fecal strains of avian E. coli produce focal infections in the avian lung, and these are accompanied by inflammation and cell death in the infected areas.

  12. Resistance Pattern and Molecular Characterization of Enterotoxigenic Escherichia coli (ETEC) Strains Isolated in Bangladesh.

    Science.gov (United States)

    Begum, Yasmin A; Talukder, K A; Azmi, Ishrat J; Shahnaij, Mohammad; Sheikh, A; Sharmin, Salma; Svennerholm, A-M; Qadri, Firdausi

    2016-01-01

    Enterotoxigenic Escherichia coli (ETEC) is a common cause of bacterial infection leading to acute watery diarrhea in infants and young children as well as in travellers to ETEC endemic countries. Ciprofloxacin is a broad-spectrum antimicrobial agent nowadays used for the treatment of diarrhea. This study aimed to characterize ciprofloxacin resistant ETEC strains isolated from diarrheal patients in Bangladesh. A total of 8580 stool specimens from diarrheal patients attending the icddr,b Dhaka hospital was screened for ETEC between 2005 and 2009. PCR and Ganglioside GM1- Enzyme Linked Immuno sorbent Assay (ELISA) was used for detection of Heat labile (LT) and Heat stable (ST) toxins of ETEC. Antimicrobial susceptibilities for commonly used antibiotics and the minimum inhibitory concentration (MIC) of nalidixic acid, ciprofloxacin and azithromycin were examined. DNA sequencing of representative ciprofloxacin resistant strains was performed to analyze mutations of the quinolone resistance-determining region of gyrA, gyrB, parC and parE. PCR was used for the detection of qnr, a plasmid mediated ciprofloxacin resistance gene. Clonal variations among ciprofloxacin resistant (CipR) and ciprofloxacin susceptible (CipS) strains were determined by Pulsed-field gel electrophoresis (PFGE). Among 1067 (12%) ETEC isolates identified, 42% produced LT/ST, 28% ST and 30% LT alone. Forty nine percent (n = 523) of the ETEC strains expressed one or more of the 13 tested colonization factors (CFs) as determined by dot blot immunoassay. Antibiotic resistance of the ETEC strains was observed as follows: ampicillin 66%, azithromycin 27%, ciprofloxacin 27%, ceftriazone 13%, cotrimaxazole 46%, doxycycline 44%, erythromycin 96%, nalidixic acid 83%, norfloxacin 27%, streptomycin 48% and tetracycline 42%. Resistance to ciprofloxacin increased from 13% in 2005 to 34% in 2009. None of the strains was resistant to mecillinam. The MIC of the nalidixic acid and ciprofloxacin of representative Cip

  13. Resistance Pattern and Molecular Characterization of Enterotoxigenic Escherichia coli (ETEC Strains Isolated in Bangladesh.

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    Yasmin A Begum

    Full Text Available Enterotoxigenic Escherichia coli (ETEC is a common cause of bacterial infection leading to acute watery diarrhea in infants and young children as well as in travellers to ETEC endemic countries. Ciprofloxacin is a broad-spectrum antimicrobial agent nowadays used for the treatment of diarrhea. This study aimed to characterize ciprofloxacin resistant ETEC strains isolated from diarrheal patients in Bangladesh.A total of 8580 stool specimens from diarrheal patients attending the icddr,b Dhaka hospital was screened for ETEC between 2005 and 2009. PCR and Ganglioside GM1- Enzyme Linked Immuno sorbent Assay (ELISA was used for detection of Heat labile (LT and Heat stable (ST toxins of ETEC. Antimicrobial susceptibilities for commonly used antibiotics and the minimum inhibitory concentration (MIC of nalidixic acid, ciprofloxacin and azithromycin were examined. DNA sequencing of representative ciprofloxacin resistant strains was performed to analyze mutations of the quinolone resistance-determining region of gyrA, gyrB, parC and parE. PCR was used for the detection of qnr, a plasmid mediated ciprofloxacin resistance gene. Clonal variations among ciprofloxacin resistant (CipR and ciprofloxacin susceptible (CipS strains were determined by Pulsed-field gel electrophoresis (PFGE.Among 1067 (12% ETEC isolates identified, 42% produced LT/ST, 28% ST and 30% LT alone. Forty nine percent (n = 523 of the ETEC strains expressed one or more of the 13 tested colonization factors (CFs as determined by dot blot immunoassay. Antibiotic resistance of the ETEC strains was observed as follows: ampicillin 66%, azithromycin 27%, ciprofloxacin 27%, ceftriazone 13%, cotrimaxazole 46%, doxycycline 44%, erythromycin 96%, nalidixic acid 83%, norfloxacin 27%, streptomycin 48% and tetracycline 42%. Resistance to ciprofloxacin increased from 13% in 2005 to 34% in 2009. None of the strains was resistant to mecillinam. The MIC of the nalidixic acid and ciprofloxacin of representative

  14. Mexican unpasteurised fresh cheeses are contaminated with Salmonella spp., non-O157 Shiga toxin producing Escherichia coli and potential uropathogenic E. coli strains: A public health risk.

    Science.gov (United States)

    Guzman-Hernandez, Rosa; Contreras-Rodriguez, Araceli; Hernandez-Velez, Rosa; Perez-Martinez, Iza; Lopez-Merino, Ahide; Zaidi, Mussaret B; Estrada-Garcia, Teresa

    2016-11-21

    Fresh cheeses are a main garnish of Mexican food. Consumption of artisanal fresh cheeses is very common and most of them are made from unpasteurised cow milk. A total of 52 fresh unpasteurised cheeses of five different types were purchased from a variety of suppliers from Tabasco, Mexico. Using the most probable number method, 67% and 63% of samples were positive for faecal coliforms and E. coli, respectively; revealing their low microbiological quality. General hygienic conditions and practices of traditional cheese manufacturers were poor; most establishments had unclean cement floors, all lacked windows and doors screens, and none of the food-handlers wore aprons, surgical masks or bouffant caps. After analysing all E. coli isolates (121 strains) for the presence of 26 virulence genes, results showed that 9 (17%) samples were contaminated with diarrheagenic E. coli strains, 8 harboured non-O157 Shiga toxin producing E. coli (STEC), and one sample contained both STEC and diffusely adherent E. coli strains. All STEC strains carried the stx1 gene. Potential uropathogenic E. coli (UPEC) strains were isolated from 15 (29%) samples; the most frequent gene combination was fimA-agn43. Two samples were contaminated with Salmonella. The results demonstrated that unpasteurised fresh cheeses produced in Tabasco are of poor microbiological quality and may frequently harbour foodborne pathogens. Food safety authorities in Mexico need to conduct more rigorous surveillance of fresh cheeses. Furthermore, simple and inexpensive measures as establishing programs emphasizing good hand milking practices and hygienic manufacturing procedures may have a major effect on improving the microbiological quality of these food items. Copyright © 2016 Elsevier B.V. All rights reserved.

  15. Pharmacodynamic modeling of in vitro activity of marbofloxacin against Escherichia coli strains.

    Science.gov (United States)

    Andraud, M; Chauvin, C; Sanders, P; Laurentie, M

    2011-02-01

    A mathematical pharmacodynamic model was developed to describe the bactericidal activity of marbofloxacin against Escherichia coli strains with reduced susceptibility levels (determined using MICs) under optimal and intestinal growth conditions. Model parameters were estimated using nonlinear least-square curve-fitting procedures for each E. coli strain. Parameters related to bactericidal activity were subsequently analyzed using a maximum-effect (E(max)) model adapted to account for a direct and a delayed effect. While net growth rates did not vary significantly with strain susceptibility, culture medium had a major effect. The bactericidal activity of marbofloxacin was closely associated with the concentration and the duration of exposure of the bacteria to the antimicrobial agent. The value of the concentration inducing a half-maximum effect (C(50)) was highly correlated with MIC values (R(2) = 0.87 and R(2) = 0.94 under intestinal and optimal conditions, respectively). Our model reproduced the time-kill kinetics with good accuracy (R(2) of >0.90) and helped explain observed regrowth.

  16. [Verification of peroral vaccination of pigs against enteropathogenic strains of Escherichia coli].

    Science.gov (United States)

    Mazura, F; Svastová, A; Kopecký, O; Salanský, P

    1982-01-01

    In a large herd of pigs where the parenteral immunisation of pregnant sows by polyvalent vaccine against enteral coliinfections of new-born piglets was performed successfully for several years, an increased occurrence of diseases and mortality of sucking piglets was observed. After screening tests in the herd, six strains of E. coli, differing from the strains contained in the commercial vaccine, were isolated. The obtained strains were employed for preparation of peroral vaccine for sows by means of the modified method after Kohler. In two cycles of sows (48 animals in experimental group and 60 animals in control group), this method was compared with the effectiveness of the commonly used commercial polyvalent parenteral vaccine against enteral E. coli infections of new-born piglets (manufactured by Bioveta, Ivanovice in Haná). In experimental sow groups vaccinated perorally by the new vaccine, the number of live-born piglets increased by 0.475 piglet per litter, the number of reared piglets up to the age of 28 days by 0.904 piglet per litter and the mortality till the age of 28 days decreased by 6.1% as compared with the control vaccinated by commercial vaccine. The results were not statistically significant. The advantages and disadvantages of both vaccination methods are discussed.

  17. Disulfide bond formation and folding of plant peroxidases expressed as inclusion body protein in Escherichia coli thioredoxin reductase negative strains

    DEFF Research Database (Denmark)

    Teilum, K; Ostergaard, L; Welinder, K G

    1999-01-01

    and the vector/host combination. The choice of E. coli strain in particular affects the yield of active peroxidase obtained in the folding step. Thus, the yield of active ATP N peroxidase can be increased 50-fold by using thioredoxin reductase negative strains, which facilitate the formation of disulfide bonds...

  18. Engineering Escherichia coli BL21(DE3) derivative strains to minimize E. coli protein contamination after purification by immobilized metal affinity chromatography.

    Science.gov (United States)

    Robichon, Carine; Luo, Jianying; Causey, Thomas B; Benner, Jack S; Samuelson, James C

    2011-07-01

    Recombinant His-tagged proteins expressed in Escherichia coli and purified by immobilized metal affinity chromatography (IMAC) are commonly coeluted with native E. coli proteins, especially if the recombinant protein is expressed at a low level. The E. coli contaminants display high affinity to divalent nickel or cobalt ions, mainly due to the presence of clustered histidine residues or biologically relevant metal binding sites. To improve the final purity of expressed His-tagged protein, we engineered E. coli BL21(DE3) expression strains in which the most recurring contaminants are either expressed with an alternative tag or mutated to decrease their affinity to divalent cations. The current study presents the design, engineering, and characterization of two E. coli BL21(DE3) derivatives, NiCo21(DE3) and NiCo22(DE3), which express the endogenous proteins SlyD, Can, ArnA, and (optionally) AceE fused at their C terminus to a chitin binding domain (CBD) and the protein GlmS, with six surface histidines replaced by alanines. We show that each E. coli CBD-tagged protein remains active and can be efficiently eliminated from an IMAC elution fraction using a chitin column flowthrough step, while the modification of GlmS results in loss of affinity for nickel-containing resin. The "NiCo" strains uniquely complement existing methods for improving the purity of recombinant His-tagged protein.

  19. Microarray analysis of the Ler regulon in enteropathogenic and enterohaemorrhagic Escherichia coli strains.

    Directory of Open Access Journals (Sweden)

    Lewis E H Bingle

    Full Text Available The type III protein secretion system is an important pathogenicity factor of enteropathogenic and enterohaemorrhagic Escherichia coli pathotypes. The genes encoding this apparatus are located on a pathogenicity island (the locus of enterocyte effacement and are transcriptionally activated by the master regulator Ler. In each pathotype Ler is also known to regulate genes located elsewhere on the chromosome, but the full extent of the Ler regulon is unclear, especially for enteropathogenic E. coli. The Ler regulon was defined for two strains of E. coli: E2348/69 (enteropathogenic and EDL933 (enterohaemorrhagic in mid and late log phases of growth by DNA microarray analysis of the transcriptomes of wild-type and ler mutant versions of each strain. In both strains the Ler regulon is focused on the locus of enterocyte effacement - all major transcriptional units of which are activated by Ler, with the sole exception of the LEE1 operon during mid-log phase growth in E2348/69. However, the Ler regulon does extend more widely and also includes unlinked pathogenicity genes: in E2348/69 more than 50 genes outside of this locus were regulated, including a number of known or potential pathogenicity determinants; in EDL933 only 4 extra-LEE genes, again including known pathogenicity factors, were activated. In E2348/69, where the Ler regulon is clearly growth phase dependent, a number of genes including the plasmid-encoded regulator operon perABC, were found to be negatively regulated by Ler. Negative regulation by Ler of PerC, itself a positive regulator of the ler promoter, suggests a negative feedback loop involving these proteins.

  20. Global gene expression profiling of the asymptomatic bacteriuria Escherichia coli strain 83972 in the human urinary tract

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Klemm, Per

    2006-01-01

    long-term bladder colonization. The strain has been used for prophylactic purposes in patients prone to more severe and recurrent UTIs. For this study, we used DNA microarrays to monitor the expression profile of strain 83972 in the human urinary tract. Significant differences in expression levels were......Urinary tract infections (UTIs) are an important health problem worldwide, with many million cases each year. Escherichia coli is the most common organism causing UTIs in humans. The asymptomatic bacteriuria E. coli strain 83972 is an excellent colonizer of the human urinary tract, where it causes...

  1. Transcriptional profiling of thymidine-producing strain recombineered from Escherichia coli BL21

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    Jin-Sook Kim

    2015-12-01

    Full Text Available DNA microarrays were used to compare the expression profiles of a thymidine overproducing strain (BLT013 and its isogenic parent, Escherichia coli BL21(DE3, when each was grown under well-defined thymidine production conditions with glycerol as carbon source. Here we describe the experimental procedures and methods in detail to reproduce the results and provide resource to be applied to similar engineering approach (available at Gene Expression Omnibus database under GSE69963. Taken together, the microarray data provide a basis for new testable hypotheses regarding enhancement of thymidine productivity and attaining a more complete understanding of nucleotide metabolism in bacteria.

  2. Respiration shutoff in Escherichia coli K12 strains is induced by far ultraviolet radiations and by mitomycin C.

    Science.gov (United States)

    Swenson, P A; Norton, I L

    1984-03-01

    Ultraviolet radiations (254 nm) (UV) cause respiration to shutoff in Escherichia coli B/r. It has been reported [P.A. Swenson, Photochem. Photobiol., 33 (1981) 855-859 and J. Barbé, A. Vericat and R. Guerrero, Mutation Res., 120 (1983) 1-5] that E. coli K12 strains do not shut off respiration after UV. The latter authors also reported that mitomycin C did not cause this 'SOS' response. In this paper we report that higher UV fluences than were previously used will cause respiration shutoff in K12 strain W3110 and that cyclic AMP increases the sensitivity of respiration shutoff of irradiated cell suspensions. We also report that mitomycin C shuts off respiration in this strain. Neither UV nor mitomycin C causes respiration shutoff in the recA56 derivative of W3110. Thus respiration shutoff is a recA dependent response to UV and mitomycin C in E. coli K12 strains.

  3. Transcriptome analysis of parallel-evolved Escherichia coli strains under ethanol stress

    Directory of Open Access Journals (Sweden)

    Yomo Tetsuya

    2010-10-01

    Full Text Available Abstract Background Understanding ethanol tolerance in microorganisms is important for the improvement of bioethanol production. Hence, we performed parallel-evolution experiments using Escherichia coli cells under ethanol stress to determine the phenotypic changes necessary for ethanol tolerance. Results After cultivation of 1,000 generations under 5% ethanol stress, we obtained 6 ethanol-tolerant strains that showed an approximately 2-fold increase in their specific growth rate in comparison with their ancestor. Expression analysis using microarrays revealed that common expression changes occurred during the adaptive evolution to the ethanol stress environment. Biosynthetic pathways of amino acids, including tryptophan, histidine, and branched-chain amino acids, were commonly up-regulated in the tolerant strains, suggesting that activating these pathways is involved in the development of ethanol tolerance. In support of this hypothesis, supplementation of isoleucine, tryptophan, and histidine to the culture medium increased the specific growth rate under ethanol stress. Furthermore, genes related to iron ion metabolism were commonly up-regulated in the tolerant strains, which suggests the change in intracellular redox state during adaptive evolution. Conclusions The common phenotypic changes in the ethanol-tolerant strains we identified could provide a fundamental basis for designing ethanol-tolerant strains for industrial purposes.

  4. Isolation and characterization of the E. coli membrane protein production strain Mutant56(DE3)

    DEFF Research Database (Denmark)

    Baumgarten, Thomas; Schlegel, Susan; Wagner, Samuel

    2017-01-01

    Membrane protein production is usually toxic to E. coli. However, using genetic screens strains can be isolated in which the toxicity of membrane protein production is reduced, thereby improving production yields. Best known examples are the C41(DE3) and C43(DE3) strains, which are both derived...... from the T7 RNA polymerase (P)-based BL21(DE3) protein production strain. In C41(DE3) and C43(DE3) mutations lowering t7rnap expression levels result in strongly reduced T7 RNAP accumulation levels. As a consequence membrane protein production stress is alleviated in the C41(DE3) and C43(DE3) strains......, thereby increasing membrane protein yields. Here, we isolated Mutant56(DE3) from BL21(DE3) using a genetic screen designed to isolate BL21(DE3)-derived strains with mutations alleviating membrane protein production stress other than the ones in C41(DE3) and C43(DE3). The defining mutation of Mutant56(DE3...

  5. Influence of Sanitizers on the Lipopolysaccharide Toxicity of Escherichia coli Strains Cultivated in the Presence of Zygosaccharomyces bailii

    Directory of Open Access Journals (Sweden)

    Lerato Mogotsi

    2014-01-01

    Full Text Available The influence of sublethal concentrations of two sanitizers, liquid iodophor and liquid hypochlorite (LH, on the growth rates and toxicity of food-borne pathogenic Escherichia coli strains grown in the presence of spoilage yeast Zygosaccharomyces bailii was assessed. When grown in combination with Z. bailii both E. coli O113 and E. coli O26 exhibited slower growth rates, except when E. coli O113 was grown in combination with Z. bailii at 0.2% LH. The growth rate of Z. bailii was not impacted by the addition of the sanitizers or by communal growth with E. coli strains. LAL and IL-6 results indicated a decrease in toxicity of pure E. coli cultures with comparable profiles for control and sanitizer exposed samples, although the LAL assay proved to be more sensitive. Interestingly, pure cultures of Z. bailii showed increased toxicity measured by LAL and decreased toxicity measured by IL-6. LAL analysis showed a decrease in toxicity of both E. coli strains grown in combination with Z. bailii, while IL-6 analysis of the mixed cultures showed an increase in toxicity. The use of LAL for toxicity determination in a mixed culture overlooks the contribution made by spoilage yeast, thus demonstrating the importance of using the appropriate method for toxicity testing in mixed microbe environments.

  6. Detection of eae, bfpA, espA Genes on Diarrhoeagenic Strains of Escherichia coli Isolates

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    Agnes Sri Harti

    2015-10-01

    Full Text Available The Enteropathogenic Escherichia coli (EPEC is one of pathogenic strain of diarrheagenic E. coli group in children andinfant that occurs in developing countries. The significant virulence factors in pathogenic EPEC are eaeA (E. coli attachingeffacing, bfpA (bundle-forming pilus A and espA (encoding secreted protein A genes. The use of DNA probes to detect thevirulence genes in E. coli in Indonesia is not common yet. In this experiment the gene fragments of eae, bfpA, and espA were usedas probes to detect the EPEC among E. coli isolates from stool specimensin of diarrheic children attending Public Health Centersin Yogyakarta. The DNA samples were isolated from 49 diarrheagenic E. coli isolates. The DNA probes of eae, bfpA and espAwere obtained by amplification of DNA fragment of EPEC O126 using PCR technique. Furthermore, those probes were used toidentify the presence of those genes among E. coli isolates using hybridization technique. The results showed that 42 (85.7%isolates were espA+, 25 isolates (51% were eaeA+ (EPEC strains. Therefore among 25 isolates of EPEC, 20 isolates (80 %among EPEC were bfpA+ (typical EPEC strains.Keywords : DNA probe, eae, bfpA, espA, EPEC.

  7. Investigation of carbon storage regulation network (csr genes) and phenotypic differences between acid sensitive and resistant Escherichia coli O157:H7 strains

    Science.gov (United States)

    Background: Escherichia coli O157:H7 and related serotype strains have previously been shown to vary in acid resistance, however, little is known about strain specific mechanisms of acid resistance. We examined sensitive and resistant E. coli strains to determine the effects of growth in minimal and...

  8. Complete genome sequences of Escherichia coli O157:H7 strains SRCC 1675 and 28RC that vary in acid resistance

    Science.gov (United States)

    The level of acid resistance among Escherichia coli O157:H7 strains varies, and strains with higher resistance to acid may have a lower infectious dose. The complete genome sequences belonging to two strains of Escherichia coli O157:H7 with different levels of acid resistance are presented....

  9. Molecular characterization of Escherichia coli O157:H7 strains isolated from different sources and geographic regions

    Science.gov (United States)

    Gonzalez, Alice Gonçalves M.; Cerqueira, Aloysio M. F.; Andrade, João Ramos C.

    2012-01-01

    Escherichia (E.) coli serotype O157:H7 is a globally distributed human enteropathogen and is comprised of microorganisms with closely related genotypes. The main reservoir for this group is bovine bowels, and infection mainly occurs after ingestion of contaminated water and food. Virulence genetic markers of 28 O157:H7 strains were investigated and multilocus enzyme electrophoresis (MLEE) was used to evaluate the clonal structure. O157:H7 strains from several countries were isolated from food, human and bovine feces. According to MLEE, O157:H7 strains clustered into two main clonal groups designated A and B. Subcluster A1 included 82% of the O157:H7 strains exhibiting identical MLEE pattern. Most enterohemorrhagic E. coli (EHEC) O157:H7 strains from Brazil and Argentina were in the same MLEE subgroup. Bovine and food strains carried virulence genes associated with EHEC pathogenicity in humans. PMID:22705735

  10. Characterization of Campylobacter jejuni and Campylobacter coli strains isolated in the region of Niš, Serbia

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    Miljković-Selimović Biljana

    2010-01-01

    Full Text Available Introduction. Campylobacter jejuni and Campylobacter coli represent one of the main causes of bacterial diarrhoea in humans. Although the disease is usually mild and self-limiting, severe chronic sequelae may occur, such as reactive arthritis, Guillain-Barré and Miller Fisher syndromes. Serotyping is used as an epidemiological marker, while post-infective polyneuropathies are associated with several O serotypes. Objective. Strains of C. jejuni and C. coli were serotyped based on heat stable (HS and heat labile (HL antigens, as well as biotypes to determine strain diversity. Methods. Campylobacter spp. was isolated using selective blood media with antibiotics. Differentiation to the species level was done by a combination of biotyping tests and by a PCR-based RFLP test. The isolates were characterised by Penner and Lior serotyping methods. Results. The serotypes showed diversity without predominant serotypes. 24 HS serotypes were detected among 29 C. jejuni strains, and seven serotypes among nine C. coli strains. HL serotyping method successfully typed 62.5% of strains. Among 16 C. jejuni strains 14 serotypes were detected, and three among four C. coli strains. A C. jejuni strain associated with a patient with Guillain-Barré syndrome was typed as biotype II, O:19. Conclusion. The biotyping and serotyping results have indicated that C. jejuni and C. coli strains in the region of Niš, Serbia are diverse and could be probably of unrelated sources of origin or reservoirs. The strain associated with the Guillain-Barré syndrome patient was serotype O:19, one of the most common in this post-infective complication.

  11. Molecular Epidemiology of ESBL Genes and Multi-Drug Resistance in Diarrheagenic Escherichia Coli Strains Isolated from Adults in Iran.

    Science.gov (United States)

    Ghorbani-Dalini, Sadegh; Kargar, Mohammad; Doosti, Abbas; Abbasi, Pejman; Sarshar, Meysam

    2015-01-01

    Resistance to oxyimino cephalosporins antibiotics in Enterobacteriaceae is primarily done by the extended spectrum β-lactamases (ESBLs). Clear identification of risk factors for ESBLs-producing infections is necessary. Therefore, efficient strategies can be developed to decrease outbreak of these infections. The aim of this study was to determine the antibacterial susceptibility and ESBLs pattern of diarrhogenic Escherichia coli (E. coli) strains isolated from adult patients. In the present study, diarrheogenic E. coli strains were isolated from 54 patients from the University of Medical Sciences hospitals in Shiraz. Antimicrobial susceptibility testing was done by disk diffusion method by CLSI criteria. The presence of bla TEM , bla SHV and bla CTX-M genes was investigated by PCR using designated primers. The prevalence of ESBLs-producer E. coli strains was 12.96%. Antimicrobial resistance testing showed a high resistance to cefexime, trimethoprim-sulfamethoxazole, ampicillin and penicillin. Overall, β-lactamase genes were identified in 52 (96.30%) isolates which were identified as 45 (83.33%) bla TEM, 17 (31.48%) blaSHV and 11 (20.37%) blaCTX-M. ESBLs-producer E. coli is very prevalent in Diarrheogenic strains isolated from adult patients. Also, this study clearly showed that the bla TEM gene for ESBLs-producer E. coli was widespread in Iran.

  12. Relative Fecal Abundance of Extended-Spectrum-β-Lactamase-Producing Escherichia coli Strains and Their Occurrence in Urinary Tract Infections in Women

    OpenAIRE

    2013-01-01

    Extended-spectrum-beta-lactamase (ESBL)-producing Escherichia coli (ESBL E. coli) strains are of major concern because few antibiotics remain active against these bacteria. We investigated the association between the fecal relative abundance (RA) of ESBL-producing E. coli (ESBL-RA) and the occurrence of ESBL E. coli urinary tract infections (UTIs). The first stool samples passed after suspicion of UTI from 310 women with subsequently confirmed E. coli UTIs were sampled and tested for ESBL-RA ...

  13. Surveillance of antimicrobial resistance in Escherichia coli strains isolated from pigs at Spanish slaughterhouses.

    Science.gov (United States)

    Teshager, T; Herrero, I A; Porrero, M C; Garde, J; Moreno, M A; Domínguez, L

    2000-07-01

    Antimicrobial resistance can make the efficient treatment of bacterial infections in humans and animals more difficult. Antimicrobial use in food animals may be one of the factors contributing to resistance. The Spanish surveillance network VAV has established a baseline of antimicrobial resistance in Escherichia coli strains from healthy pigs. Minimum inhibitory concentration and patterns of resistance to antimicrobials used in animals and humans were determined for 205 faecal strains isolated in a sampling frame of four slaughterhouses in Spain from 220 pigs in 1998. Higher levels of resistance were seen against antimicrobial agents authorised for use in food animals especially tetracycline, sulphonamides, trimethoprim and amoxycillin. All isolates were susceptible to antimicrobials employed mainly in humans such as ceftazidime, cefotaxime, imipenem, aztreonam and amikacin.

  14. A survey of antibiotic resistance among E. coli strains isolated from poultry in Karachi.

    Science.gov (United States)

    Ansari, F A; Khatoon, H

    1999-01-01

    Studies were carried out to investigate the incidence of multiple antibiotic resistance among E .coli (total 152) isolated from poultry in Karachi to eight commonly used antibiotics: ampicillin (A), chloramphenicol (C), gentamycin (G), anamycin (K), neomycin (N), polymyxin B (P), streptomycin (S) and tetracycline (T) at the levels of 50 microg/ml, 100 microg/ml and 500 microg/ml. Tables of the results are given, showing the number of resistant strains of different patterns of antibiotic resistance at different levels. A comparison of antibiotic resistance to different number of antibiotics and the frequency of resistance to individual antibiotic at different levels is also reported. The highest frequency of resistance was against tetracycline whereas the lowest frequency of resistance was against gentamycin. Thirty R plasmids were isolated from the resistant strains and will be reported elsewhere.

  15. Molecular mechanisms that mediate colonization of Shiga toxin-producing Escherichia coli strains.

    Science.gov (United States)

    Farfan, Mauricio J; Torres, Alfredo G

    2012-03-01

    Shiga toxin-producing Escherichia coli (STEC) is a group of pathogens which cause gastrointestinal disease in humans and have been associated with numerous food-borne outbreaks worldwide. The intimin adhesin has been considered for many years to be the only colonization factor in these strains. However, the rapid progress in whole-genome sequencing of different STEC serotypes has accelerated the discovery of other adhesins (fimbrial and afimbrial), which have emerged as important contributors to the intestinal colonization occurring during STEC infection. This review summarizes recent progress to identify and characterize, at the molecular level, novel adhesion and colonization factors in STEC strains, with an emphasis on their contribution to virulence traits, their host-pathogen interactions, the regulatory mechanisms controlling their expression, and their role as targets eliciting immune responses in the host.

  16. Quantitative evaluation of extended-spectrum β-lactamase-producing Escherichia coli strains in the wastewater of a French teaching hospital and relation to patient strain

    Directory of Open Access Journals (Sweden)

    Laurence Drieux

    2016-03-01

    Full Text Available Abstract Background Extended-spectrum β-lactamase-producing Escherichia coli has become ubiquitous and has been reported in diverse ecosystems. We evaluated the potential impact of post-acute and long-term healthcare activities on the environment by quantifying ESBL-producing Enterobacteriaceae in wastewaters of a French geriatric hospital. Methods We collected wastewater specimens representative of one-day efflux immediately before the connection with the municipal sewer pipe. The sample was processed following two different methods: dilution-filtration method and concentration method and was screened for ESBL-producing Enterobacteriaceae using selective media. ESBL E. coli strains were quantified, screened for ESBL genes and compared with ESBL strains isolated from patients present in the building at the time of wastewater collection, using molecular methods. Results Six distinct environmental ESBL E. coli clusters were identified, two of them related to patient strains. The concentrations in hospital wastewater of these strains ranged from 2.5 × 104 to 106 UFC/L. Conclusions Our results demonstrate that the presence of ESBL E. coli patients leads to a dissemination of ESBL E. coli in the environment and highlights the need to improve excreta and wastewater policy in hospitals.

  17. Investigations of multiresistance to antibiotics and chemotherapeutics and extended spectrum beta: Lactamase effect (ESBL test in strains E.coli and salmonella originating from domestic animals

    Directory of Open Access Journals (Sweden)

    Mišić Dušan

    2006-01-01

    Full Text Available The presence of multiresistance to the effects of antibiotics and chemotherapeutics and extended spectrum beta-lactamase were examined in 45 strains of E. coli and 35 strains of Salmonella. The strains of E. coli originated from several species of domestic animals: dogs, cats, poultry, and cattle, and 30 strains of Salmonella originated from poultry, 4 strains from cattle, and 1 strain from swine. The presence of the following serovarieties was established using serological examinations: Salmonella Enteritidis 17 strains, Salmonella Gallinarum 1 strain, Salmonella Hartford 5 strains, Salmonella Anatum 1 strain, Salmonella Typhimurium 4 strains, Salmonella Agona 1 strain, Salmonella Infantis 1 strain, Salmonella Thompson var. Berlin 1 strain, Salmonella Tennessee 1 strain, Salmonella Senftenberg 1 strain, Salmonella Glostrup 1 strain, and Salmonella Hadar 1 strain. In the examinations of the listed strains we used antibiogram discs of ampicillin, amoxicillin with clavulanic acid, cephalexin, cephtriaxon, cephotaxim, cephtazidime, aztreonam, gentamycin, chloramphenicol, tetracycline, cyprofloxacine, and a combination of sulphamethoxasole and trimethoprim. The lowest prevalence of multiresistance in E. Coli strains to 3 or more antibiotics was established in dogs 20%, and the highest in 60% strains originating from swine. In 62.88% strains of Salmonella we established sensitivity to all applied antibiotics. Resistance was also established in a small number of the examined strains to ampicillin (11 strains, to tetracycline (5 strains, to amoxicillin with clavulanic acid (5 strains, to sulphamethoxasole with trimethoprim (5 strains, to gentamycin (3 strains, and to cloramphenicol (1 strain. Of all the examined strains of Salmonella, 6 strains originating from poultry exhibited multiresistence. The presence of extended spectrum beta-lactamase effects examined using the ESBL test, was not established in strains of E. coli and Salmonella strains.

  18. Molecular typing of Escherichia coli strains associated with threatened sea ducks and near-shore marine habitats of southwest Alaska

    Science.gov (United States)

    Schamber, Jason L.

    2011-01-01

    In Alaska, sea ducks winter in coastal habitats at remote, non-industrialized areas, as well as in proximity to human communities and industrial activity. We evaluated prevalence and characteristics of Escherichia coli strains in faecal samples of Steller's eiders (Polysticta stelleri; n = 122) and harlequin ducks (Histrionicus histrionicus; n = 21) at an industrialized site and Steller's eiders (n = 48) at a reference site, and compared these strains with those isolated from water samples from near-shore habitats of ducks. The overall prevalence of E. coli was 16% and 67% in Steller's eiders and harlequin ducks, respectively, at the industrialized study site, and 2% in Steller's eiders at the reference site. Based on O and H antigen subtyping and genetic characterization by enterobacterial repetitive intergenic consensus polymerase chain reaction and pulsed-field gel electrophoresis, we found evidence of avian pathogenic E. coli (APEC) strains associated with both species and detected E. coli strains carrying virulence genes associated with mammals in harlequin ducks. Steller's eiders that carried APEC had lower serum total protein and albumin concentrations, providing further evidence of pathogenicity. The genetic profile of two E. coli strains from water matched an isolate from a Steller's eider providing evidence of transmission between near-shore habitats and birds.

  19. Association of IL-8-inducing strains of diffusely adherent Escherichia coli with sporadic diarrheal patients with less than 5 years of age

    OpenAIRE

    Ismail Mustafa Meraz; Kentaro Arikawa; Hiromi Nakamura; Jun Ogasawara; Atsushi Hase; Yoshikazu Nishikawa

    2007-01-01

    The role of diffusely adherent Escherichia coli (DAEC) in diarrheal disease has been controversial. However, DAEC strains were recently implicated in diarrheal disease in developing countries. To clarify whether DAEC are prevalent among sporadic cases of diarrheal illness in Osaka City, Japan, E. coli strains isolated between July 1997 and March 2000 during diarrheagenic E. coli (DEC) investigation were retrospectively examined. DAEC strains were recognized among 41 (4.4%) of 924 patients and...

  20. [Investigation of plasmid-mediated quinolone resistance in Escherichia coli strains].

    Science.gov (United States)

    Aktepe, Orhan Cem; Aşık, Gülşah; Cetinkol, Yeliz; Biçmen, Meral; Gülay, Zeynep

    2012-01-01

    Quinolones are widely used antimicrobial agents, particularly for the treatment of infections caused by gram-negative bacilli such as E.coli. As a consequence, quinolone resistance has been increasing among this species in recent years. Bacterial resistance to quinolones usually results from mutations in the chromosomal genes which encode topoisomerases and also the expression of efflux pumps and loss of porines contributed to development of quinolone resistance. However, recent studies have shown that the spread and increase of quinolone resistance may be due to the transfer of plasmid-mediated genes. To date, three groups of plasmid-mediated quinolone resistance genes, namely qnr, aac(6')-Ib-cr, and qepA, have been described. The aim of this study was to investigate the presence of plasmid-mediated quinolone resistance genes in E.coli clinical isolates. A total of 112 quinolone-resistant E.coli strains isolated from different clinical specimens (84 urine, 16 blood, 10 wound, 2 bronchoalveolar lavage) of which 78 (69.6%) were extended-spectrum beta-lactamase (ESBL) positive, in Afyon Kocatepe University Hospital, Microbiology Laboratory were included in the study. In the isolates, qnrA, qnrB, qnrS, qnrC, qepA, and aac(6')-1b-cr plasmid genes were analysed by polymerase chain reaction (PCR). After aac(6')- 1b determinant was amplified by PCR, all aac(6')-1b positive amplicons were analyzed by digestion with BseGI restriction enzyme to identify aac(6')-1b-cr variant. It was found that, none of the strains horboured qnrA, qnrB, qnrS, qnrC and qepA genes, however, plasmid-mediated quinolone resistance gene aac(6')-1b-cr was found positive in 59.8% (67/112) of the strains. It was notable that 86.6% (58/67) of those isolates were ESBL producers. The rates of quinolone resistance among E.coli isolates infections were high in our region and an increasing trend has been observed in recent years. Our data indicated that the presence of plasmid- mediated resistance genes

  1. Detection of virulence genes in Uropathogenic E. coli (UPEC strains by Multiplex-PCR method

    Directory of Open Access Journals (Sweden)

    Javad Mohammadi

    2017-06-01

    Full Text Available Background & Objectives: Urinary tract infection caused by E. coli is one of the most common illnesses in all age groups worldwide. Presence of virulence genes is a key factor in bacterial pathogens in uroepithelial cells. The present study was performed to detect iha, iroN, ompT genes in the Uropathogenic E.coli isolates from clinical samples using multiplex-PCR method in Kerman. Materials & Methods: In this descriptive cross-sectional study, 200 samples of patients with urinary tract infections in Kerman hospitals were collected. After biochemical and microbiological tests, all strains were tested with regard to the presence of iha, iroN, and ompT genes using multiplex-PCR method. Results: The results of Multiplex-PCR showed that all specimens had one, two, or three virulence genes simultaneously. The highest and lowest frequency distribution of genes was related to iha (56.7% and iroN (20% respectively. Conclusion: According to the prevalence of urinary tract infection in the community and distribution of resistance and virulence factors, the fast and accurate detection of the strains and virulence genes is necessary

  2. Enhanced ethanol fermentation of brewery wastewater using the genetically modified strain E. coli KO11.

    Science.gov (United States)

    Rao, Kripa; Chaudhari, Vaibhav; Varanasi, Sasidhar; Kim, Dong-Shik

    2007-02-01

    We have used liquid waste obtained from a beer brewery process to produce ethanol. To increase the productivity, genetically modified organism, Escherichia coli KO11, was used for ethanol fermentation. Yeast was also used to produce ethanol from the same feed stock, and the ethanol production rates and resulting concentrations of sugars and ethanol were compared with those of KO11. In the experiments, first the raw wastewater was directly fermented using two strains with no saccharification enzymes added. Then, commercial enzymes, alpha-amylase, pectinase, or a combination of both, were used for simultaneous saccharification and fermentation, and the results were compared with those of the no-enzyme experiments for KO11 and yeast. Under the given conditions with or without the enzymes, yeast produced ethanol more rapidly than E. coli KO11, but the final ethanol concentrations were almost the same. For both yeast and KO11, the enzymes were observed to enhance the ethanol yields by 61-84% as compared to the fermentation without enzymes. The combination of the two enzymes increased ethanol production the most for the both strains. The advantages of using KO11 were not demonstrated clearly as compared to the yeast fermentation results.

  3. Hydrogen production and metabolic flux analysis of metabolically engineered Escherichia coli strains

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Seohyoung; Seol, Eunhee; Park, Sunghoon [Department of Chemical and Biochemical Engineering, Pusan National University, Busan 609-735 (Korea); Oh, You-Kwan [Bioenergy Research Center, Korea Institute of Energy Research, Daejeon 305-543 (Korea); Wang, G.Y. [Department of Oceanography, University of Hawaii at Manoa Honolulu, HI 96822 (United States)

    2009-09-15

    Escherichia coli can produce H{sub 2} from glucose via formate hydrogen lyase (FHL). In order to improve the H{sub 2} production rate and yield, metabolically engineered E. coli strains, which included pathway alterations in their H{sub 2} production and central carbon metabolism, were developed and characterized by batch experiments and metabolic flux analysis. Deletion of hycA, a negative regulator for FHL, resulted in twofold increase of FHL activity. Deletion of two uptake hydrogenases (1 (hya) and hydrogenase 2 (hyb)) increased H{sub 2} production yield from 1.20 mol/mol glucose to 1.48 mol/mol glucose. Deletion of lactate dehydrogenase (ldhA) and fumarate reductase (frdAB) further improved the H{sub 2} yield; 1.80 mol/mol glucose under high H{sub 2} pressure or 2.11 mol/mol glucose under reduced H{sub 2} pressure. Several batch experiments at varying concentrations of glucose (2.5-10 g/L) and yeast extract (0.3 or 3.0 g/L) were conducted for the strain containing all these genetic alternations, and their carbon and energy balances were analyzed. The metabolic flux analysis revealed that deletion of ldhA and frdAB directed most of the carbons from glucose to the glycolytic pathway leading to H{sub 2} production by FHL, not to the pentose phosphate pathway. (author)

  4. ASSESMENT OF CRYOPRESERVATION SYSTEMS INFLUENCE ON THE SURVAVIAL OF E. COLI RECOMBINANT STRAINS

    Directory of Open Access Journals (Sweden)

    CLAUDIA TEREZIA SOCOL

    2013-12-01

    Full Text Available The cryopreservation systems of recombinant bacterial cells based on glycerol were studied in these experiments according to the hypothesis that glycerol is one of the widely used cryoprotective additives in microbiology and a multitude of factors affecting the effectiveness of cryopreservation in microorganisms; the best cryoprotective additive and the optimum concentration for a particular microorganism has to be determined empirically. The results obtained in this experiment are showing that the freezing procedure at -80°C in LB 40% glycerol is the optimum system for the cryopreservation of E. coli DH5α recombinant cells. The use of SOC medium supplemented with 10g/l NaCl provided more proper conditions of culture for the defrosted E. coli DH5α recombinant cells, reducing the osmotic stress during the recovery after thawing. The utilization of this optimum cryopreservation system offer the possibility of preserving the large volume of work and time involved by the recombinant DNA technology procedures applied for obtaining a recombinant strain, avoiding the storage of recombinant strains by costly and time consuming microbiology culturing techniques.

  5. An indole-deficient Escherichia coli strain improves screening of cytochromes P450 for biotechnological applications.

    Science.gov (United States)

    Brixius-Anderko, Simone; Hannemann, Frank; Ringle, Michael; Khatri, Yogan; Bernhardt, Rita

    2017-05-01

    Escherichia coli has developed into an attractive organism for heterologous cytochrome P450 production, but, in some cases, was restricted as a host in view of a screening of orphan cytochromes P450 or mutant libraries in the context of molecular evolution due to the formation of the cytochrome P450 inhibitor indole by the enzyme tryptophanase (TnaA). To overcome this effect, we disrupted the tnaA gene locus of E. coli C43(DE3) and evaluated the new strain for whole-cell substrate conversions with three indole-sensitive cytochromes P450, myxobacterial CYP264A1, and CYP109D1 as well as bovine steroidogenic CYP21A2. For purified CYP264A1 and CYP21A2, the half maximal inhibitory indole concentration was determined to be 140 and 500 μM, which is within the physiological concentration range occurring during cultivation of E. coli in complex medium. Biotransformations with C43(DE3)_∆tnaA achieved a 30% higher product formation in the case of CYP21A2 and an even fourfold increase with CYP264A1 compared with C43(DE3) cells. In whole-cell conversion based on CYP109D1, which converts indole to indigo, we could successfully avoid this reaction. Results in microplate format indicate that our newly designed strain is a suitable host for a fast and efficient screening of indole-influenced cytochromes P450 in complex medium. © 2016 International Union of Biochemistry and Molecular Biology, Inc.

  6. Draft Genome Sequence of Extended-Spectrum-β-Lactamase-Producing Escherichia coli Strain CCUG 62462, Isolated from a Urine Sample

    Science.gov (United States)

    Johnning, Anna; Jakobsson, Hedvig E.; Boulund, Fredrik; Salvà-Serra, Francisco; Åhrén, Christina; Kristiansson, Erik

    2016-01-01

    The draft genome sequence has been determined for an extended-spectrum-β-lactamase (ESBL)-producing (blaCTX-M-15) Escherichia coli strain (CCUG 62462), composed of 119 contigs and a total size of 5.27 Mb. This E. coli is serotype O25b and sequence type 131, a pandemic clonal group, causing worldwide antimicrobial-resistant infections. PMID:27979938

  7. Exploring the proteomic characteristics of the Escherichia coli B and K-12 strains in different cellular compartments.

    Science.gov (United States)

    Han, Mee-Jung

    2016-07-01

    Escherichia coli, one of the well-characterized prokaryotes, has been the most widely used bacterial host in scientific studies and industrial applications. Many different strains have been developed for the widespread use of E. coli in biotechnology, and selecting an ideal host to produce a specific protein of interest is a critical step in developing a production process. The E. coli B and K-12 strains are among the most frequently used bacterial hosts for the production of recombinant proteins as well as small-molecule metabolites such as amino acids, biofuels, carboxylic acids, diamines, and others. However, both strains have distinctive differences in genotypic and phenotypic attributes, and their behaviors can still be unpredictable at times, especially while expressing a recombinant protein. Therefore, in this review, an in-depth analysis of the physiological behavior on the proteomic level was performed, wherein the particularly distinct proteomic differences between the E. coli B and K-12 strains were investigated in the four distinctive cellular compartments. Interesting differences in the proteins associated with key cellular properties including cell growth, protein production and quality, cellular tolerance, and motility were observed between the two representative strains. The resulting enhancement of knowledge regarding host physiology that is summarized herein is expected to contribute to the acceleration of strain improvements and optimization for biotechnology-related processes.

  8. Pathogenicity of Vietnamese enterotoxigenic Escherichia coli strains in colostrum-deprived one-day-old piglets.

    Science.gov (United States)

    Do, T N; Wilkie, I; Driesen, S J; Fahy, V A; Trott, D J

    2006-03-01

    Preweaning colibacillosis is a major cause of economic loss to the swine industry in Vietnam. The aim of this study was to examine the enteropathogenicity of representative enterotoxigenic Escherichia coli (ETEC) strains obtained during an earlier epidemiologic survey conducted in five provinces in North Vietnam. This included isolates belonging to serotype O8 that produced heat-stable and heat-labile enterotoxins but did not produce any of the recognized fimbriae (F4, F5, F6, F41, F18). In vitro hemagglutination (unique mannose-resistant hemagglutination activity with guinea pig, sheep, human, and chicken red blood cells at 37 degrees C, but not at 18 degrees C) and enterocyte brush border attachment assays suggested that the F- ETEC strains produced an unidentified colonization factor that promoted adherence to the intestinal epithelium. Colostrum-deprived 1-day-old piglets challenged with an F- strain (1-2 x 10(9) bacteria) developed acute watery diarrhea within 4 hours of inoculation and suffered up to 20% weight loss, with comparable severity to piglets challenged with conventional F4 and F5 strains. At necropsy, viable counts and histopathologic examination of intestinal sections demonstrated colonization of the duodenum, jejunum, and ileum by F4-positive strains. In comparison, the F- and F5-positive strains attached exclusively to the ileum. Transmission electron micrographs of negatively stained F- cells grown at 37 degrees C demonstrated the presence of fimbriae. These results confirm the presence of a potentially new pathogenic ETEC fimbrial type in piggeries in Vietnam, with a unique hemagglutination property and attachment characteristics similar to ETEC bearing F5 fimbriae.

  9. Comparison of antimicrobial resistance in Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes strains isolated from organic and conventional poultry meat.

    Science.gov (United States)

    Miranda, J M; Vázquez, B I; Fente, C A; Calo-Mata, P; Cepeda, A; Franco, C M

    2008-12-01

    The presence of Escherichia coli, Staphylococcus aureus, and Listeria monocytogenes was determined in 55 samples of organic poultry meat and in 61 samples of conventional poultry meat. A total of 220 E. coli, 192 S. aureus, and 71 L. monocytogenes strains were analyzed by an agar disk diffusion assay for their resistance to ampicillin, cephalothin, chloramphenicol, ciprofloxacin, doxycycline, fosfomycin, gentamicin, nitrofurantoin, streptomycin, and sulfisoxazole (E. coli); chloramphenicol, ciprofloxacin, clindamycin, doxycycline, erythromycin, gentamicin, nitrofurantoin, oxacillin, and sulfisoxazole (S. aureus); and chloramphenicol, doxycycline, erythromycin, gentamicin, sulfisoxazole, and vancomycin (L. monocytogenes). The results indicated a significantly higher (P poultry meat as compared with conventional poultry meat. E. coli isolated from organic poultry meat exhibited lower levels of antimicrobial resistance against 7 of the 10 antimicrobials tested as compared with isolates recovered from conventional meat. In the case of S. aureus and L. monocytogenes isolated from conventional poultry, antimicrobial resistance was significantly higher only for doxycycline as compared with strains isolated from organic poultry. In the case of E. coli, the presence of multiresistant strains was significantly higher (P poultry meat as compared with organic poultry meat. Organically farmed poultry samples showed significantly lower development of antimicrobial resistance in intestinal bacteria such as E. coli.

  10. Cross feeding of glucose metabolism byproducts of Escherichia coli human gut isolates and probiotic strains affect survival of Vibrio cholerae.

    Science.gov (United States)

    Sengupta, Chirantana; Ekka, Manjula; Arora, Saurabh; Dhaware, Prashant D; Chowdhury, Rukhsana; Raychaudhuri, Saumya

    2017-01-01

    Vibrio cholerae converts glucose into either acid or the neutral end product acetoin and its survival in carbohydrate enriched media is linked to the nature of the byproducts produced. It has been demonstrated in this study that Escherichia coli strain isolated from the gut of healthy human volunteers and the commonly used probiotic E. coli Nissle strain that metabolize glucose to acidic byproducts drastically reduce the survival of V. cholerae strains irrespective of their glucose sensitivity and acetoin production status. Accordingly, E. coli glucose transport mutants that produce lower amounts of acidic metabolites had little effect on the survival of V. cholerae in cocultures. Thus, cross feeding of byproducts of glucose metabolism by heterologous bacteria modulates the survival of V. cholerae in glucose rich medium suggesting that composition of the gut microbiota could influence the outcome of V. cholerae infection especially when glucose based ORS is administered.

  11. Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks

    Directory of Open Access Journals (Sweden)

    M.C. Fernandes

    2013-01-01

    Full Text Available The identification of pathogens of viral (Rotavirus, Coronavirus, parasitic (Toxocara spp. and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5% strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene or intermediately virulent (vapB gene profiles. Sixty-three (31.5% strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%, A (25.4% and B2 (22.2%. No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

  12. Identification of pathogens and virulence profile of Rhodococcus equi and Escherichia coli strains obtained from sand of parks.

    Science.gov (United States)

    Fernandes, M C; Takai, S; Leite, D S; Pinto, J P A N; Brandão, P E; Santarém, V A; Listoni, F J P; Da Silva, A V; Ribeiro, M G

    2013-01-01

    The identification of pathogens of viral (Rotavirus, Coronavirus), parasitic (Toxocara spp.) and bacterial (Escherichia coli, Salmonella spp., Rhodococcus equi) origin shed in feces, and the virulence profile of R. equi and E. coli isolates were investigated in 200 samples of sand obtained from 40 parks, located in central region of state of Sao Paulo, Brazil, using different diagnostic methods. From 200 samples analyzed, 23 (11.5%) strains of R. equi were isolated. None of the R. equi isolates showed a virulent (vapA gene) or intermediately virulent (vapB gene) profiles. Sixty-three (31.5%) strains of E. coli were identified. The following genes encoding virulence factors were identified in E. coli: eae, bfp, saa, iucD, papGI, sfa and hly. Phylogenetic classification showed that 63 E. coli isolates belonged to groups B1 (52.4%), A (25.4%) and B2 (22.2%). No E. coli serotype O157:H7 was identified. Eggs of Toxocara sp. were found in three parks and genetic material of bovine Coronavirus was identified in one sample of one park. No Salmonella spp. and Rotavirus isolates were identified in the samples of sand. The presence of R. equi, Toxocara sp, bovine Coronavirus and virulent E. coli isolates in the environment of parks indicates that the sanitary conditions of the sand should be improved in order to reduce the risks of fecal transmission of pathogens of zoonotic potential to humans in these places.

  13. Global Expression of Prophage Genes in Escherichia coli O157:H7 Strain EDL933 in Response to Norfloxacin

    Science.gov (United States)

    Herold, Sylvia; Siebert, Jutta; Huber, Andrea; Schmidt, Herbert

    2005-01-01

    We investigated the influence of a low concentration of the gyrase inhibitor norfloxacin on the transcriptome of enterohemorrhagic Escherichia coli O157:H7 strain EDL933. For this purpose, we used a commercial DNA microarray containing oligonucleotides specific for E. coli O157:H7 strains EDL933 and RIMD0509952 and E. coli K-12 strain MG1655. Under the conditions applied, 5,963 spots (94% of all spots) could be analyzed. Among these, 118 spots (P norfloxacin. Twenty-two additional upregulated genes appeared to be E. coli O157:H7 strain RIMD0509952-specific phage elements, and the remaining 11 genes were related mainly to recombination and stress functions. Downregulation was indicated predominantly for genes responsible for bacterial primary metabolism, such as energy production, cell division, and amino acid biosynthesis. Interestingly, some genes present in the locus of enterocyte effacement appeared to be downregulated. The results of the study have shown that a low concentration of norfloxacin has profound effects on the transcriptome of E. coli O157:H7. PMID:15728886

  14. Variability of Escherichia coli O157 Strain Survival in Manure-Amended Soil in Relation to Strain Origin, Virulence Profile, and Carbon Nutrition Profile▿†

    Science.gov (United States)

    Franz, Eelco; van Hoek, Angela H. A. M.; Bouw, El; Aarts, Henk J. M.

    2011-01-01

    The variation in manure-amended soil survival capability among 18 Escherichia coli O157 strains (8 animal, 1 food, and 9 human isolates) was studied using a single sandy soil sample and a single sample of cattle manure as the inoculum carrier. The virulence profiles of E. coli O157 strains were characterized by detection of virulence determinants (73 genes, 122 probes in duplicate) by using the Identibac E. coli genotyping DNA miniaturized microarray. Metabolic profiling was done by subjecting all strains to the Biolog phenotypic carbon microarray. Survival times (calculated as days needed to reach the detection limit using the Weibull model) ranged from 47 to 266 days (median, 120 days). Survival time was significantly higher for the group of human isolates (median, 211 days; minimum [min.], 71; maximum [max.], 266) compared to the group of animal isolates (median, 70 days; min., 47; max., 249) (P = 0.025). Although clustering of human versus animal strains was observed based on pulsed-field gel electrophoresis (PFGE) patterns, no relation between survival time and the presence of virulence genes was observed. Principal component analysis on the metabolic profiling data revealed distinct clustering of short- and long-surviving strains. The oxidization rate of propionic acid, α-ketobutyric acid, and α-hydroxybutyric acid was significantly higher for the long-surviving strains than for the short-surviving strains. The oxidative capacity of E. coli O157 strains may be regarded as a phenotypic marker for enhanced survival in manure-amended soil. The large variation observed in survival is of importance for risk assessment models. PMID:21908630

  15. Colonization of the respiratory tract by a virulent strain of avian Escherichia coli requires carriage of a conjugative plasmid.

    Science.gov (United States)

    Ginns, C A; Benham, M L; Adams, L M; Whithear, K G; Bettelheim, K A; Crabb, B S; Browning, G F

    2000-03-01

    The E3 strain of E. coli was isolated in an outbreak of respiratory disease in broiler chickens, and experimental aerosol exposure of chickens to this strain induced disease similar to that seen in the field. In order to establish whether the virulent phenotype of this strain was associated with carriage of particular plasmids, four plasmid-cured derivatives, each lacking two or more of the plasmids carried by the wild-type strain, were assessed for virulence. Virulence was found to be associated with one large plasmid, pVM01. Plasmid pVM01 was marked by introduction of the transposon TnphoA, carrying kanamycin resistance, and was then cloned by transformation of E. coli strain DH5alpha. The cloned plasmid was then reintroduced by conjugation into an avirulent plasmid-cured derivative of strain E3 which lacked pVM01. The conjugant was shown to be as virulent as the wild-type strain E3, establishing that this plasmid is required for virulence following aerosol exposure. This virulence plasmid conferred expression of a hydroxamate siderophore, but not colicins, on both strain E3 and strain DH5alpha. Carriage of this plasmid was required for strain E3 to colonize the respiratory tracts of chickens but was not necessary for colonization of the gastrointestinal tract. However, the virulence plasmid did not confer virulence, or the capacity to colonize the respiratory tract, on strain DH5alpha. Thus, these studies have established that infection of chickens with E. coli strain E3 by the respiratory route is dependent on carriage of a conjugative virulence plasmid, which confers the capacity to colonize specifically the respiratory tract and which also carries genes for expression of a hydroxymate siderophore. These findings will facilitate identification of the specific genes required for virulence in these pathogens.

  16. SHuffle, a novel Escherichia coli protein expression strain capable of correctly folding disulfide bonded proteins in its cytoplasm

    Directory of Open Access Journals (Sweden)

    Lobstein Julie

    2012-05-01

    Full Text Available Abstract Background Production of correctly disulfide bonded proteins to high yields remains a challenge. Recombinant protein expression in Escherichia coli is the popular choice, especially within the research community. While there is an ever growing demand for new expression strains, few strains are dedicated to post-translational modifications, such as disulfide bond formation. Thus, new protein expression strains must be engineered and the parameters involved in producing disulfide bonded proteins must be understood. Results We have engineered a new E. coli protein expression strain named SHuffle, dedicated to producing correctly disulfide bonded active proteins to high yields within its cytoplasm. This strain is based on the trxB gor suppressor strain SMG96 where its cytoplasmic reductive pathways have been diminished, allowing for the formation of disulfide bonds in the cytoplasm. We have further engineered a major improvement by integrating into its chromosome a signal sequenceless disulfide bond isomerase, DsbC. We probed the redox state of DsbC in the oxidizing cytoplasm and evaluated its role in assisting the formation of correctly folded multi-disulfide bonded proteins. We optimized protein expression conditions, varying temperature, induction conditions, strain background and the co-expression of various helper proteins. We found that temperature has the biggest impact on improving yields and that the E. coli B strain background of this strain was superior to the K12 version. We also discovered that auto-expression of substrate target proteins using this strain resulted in higher yields of active pure protein. Finally, we found that co-expression of mutant thioredoxins and PDI homologs improved yields of various substrate proteins. Conclusions This work is the first extensive characterization of the trxB gor suppressor strain. The results presented should help researchers design the appropriate protein expression conditions using

  17. Comprehensive expression analysis of pathogenicity genes in uropathogenic Escherichia coli strains.

    Science.gov (United States)

    Paniagua-Contreras, Gloria Luz; Hernández-Jaimes, Tania; Monroy-Pérez, Eric; Vaca-Paniagua, Felipe; Díaz-Velásquez, Clara; Uribe-García, Alina; Vaca, Sergio

    2017-02-01

    In this study, we investigated distinct expression patterns of genes encoding iron-acquisition systems, adhesins, protectins, and toxins in human uroepithelial cells infected with 194 uropathogenic Escherichia coli (UPEC) strains in vitro. We assessed the association of these genes with antibiotic resistance genes in this group of UPEC strains, previously characterised by polymerase chain reaction (PCR). Strains were isolated from patients with urinary tract infections (UTIs) from Unidad Médica Familiar de Salud Pública, located in Estado de México, México. Antibiotic resistance genes were identified by PCR, and the expression of virulence genes was detected by reverse-transcriptase-PCR after in vitro infection of cultured A431 human keratinocytes derived from a vulvar epidermoid carcinoma. The most frequently expressed virulence genotypes among the investigated UPEC strains included usp (68%), iha (64.9%), kpsMT (61.3%), fim (58.2%), irp2 (48.4), papC (33.5%), set (31.4%) and astA (30.9%), whereas the most frequently detected antibiotic resistance genes were tet(A) (34%), sul1 (31.4%) and TEM (26.3%). Furthermore, the most abundant pattern of gene expression (irp2/fim/iha/kpsMT/usp), associated with 8 different combinations of antibiotic resistance genotypes, was exhibited by 28 strains (14.4%). Taken together, these results indicate collective participation of distinct virulence UPEC genotypes during in vitro infection of cultured human epithelial cells, suggesting their potential involvement in UTI pathogenesis. Copyright © 2016 Elsevier Ltd. All rights reserved.

  18. [Pathogenic role of strains of specific Escherichia coli serotypes in the development of mastitis in cows. I. Clinical course].

    Science.gov (United States)

    Malinowski, E

    1984-01-01

    The studies were carried out on 14 cows. For infection of the cows pathogenic (from the field) E. coli strains from the serological groups: 093, 010,086 and non pathogenic strains (museum) belonging to serogroups: 0132, 0120, 093 were used. The microorganisms of each group were injected into the udders of two cows. Two successive cows were injected into the udders E. coli endotoxin (group 093). In 6 hr after infusion of the strain or endotoxin, the experimental quarters were swollen, reddened, became hot, painful and hard. The internal temperature values (40,9-41,7 degrees C), pulse rate (100-120) and breathing rate (over 40) rapidly increased. The animals stopped taking food and ruminating. There appeared convulsions, serous discharges from the nose and eyes, sialorrhea, diarrhea. Then the general symptoms disappeared after 9 hr in cows which received endotoxin, and after 12 hr in cows infected with the museum strains. However, the cows infected with the field E. coli strains showed a very weak appetite for 2-4 days, despite the temperature dropped. After the general symptoms disappeared, changes in the experimental udder quarters were intensified and then they were observed to retreat. During 10 days the disease symptoms disappeared in the cows which were given endotoxin. These symptoms persisted longer (up to 14 days) in cows infected with the museum strains. However, in cows infected with the field strains (particularly from group 093) the inflammatory state in the udder lasted till the end of 14-day observation.

  19. A versatile Escherichia coli strain for identification of biotin transporters and for biotin quantification

    Science.gov (United States)

    Finkenwirth, Friedrich; Kirsch, Franziska; Eitinger, Thomas

    2014-01-01

    Biotin is an essential cofactor of carboxylase enzymes in all kingdoms of life. The vitamin is produced by many prokaryotes, certain fungi, and plants. Animals depend on biotin uptake from their diet and in humans lack of the vitamin is associated with serious disorders. Many aspects of biotin metabolism, uptake, and intracellular transport remain to be elucidated. In order to characterize the activity of novel biotin transporters by a sensitive assay, an Escherichia coli strain lacking both biotin synthesis and its endogenous high-affinity biotin importer was constructed. This strain requires artificially high biotin concentrations for growth. When only trace levels of biotin are available, it is viable only if equipped with a heterologous high-affinity biotin transporter. This feature was used to ascribe transport activity to members of the BioY protein family in previous work. Here we show that this strain together with its parent is also useful as a diagnostic tool for wide-concentration-range bioassays. PMID:24256712

  20. Clonal analysis of Escherichia coli serotype O6 strains from urinary tract infections.

    Science.gov (United States)

    Zingler, G; Ott, M; Blum, G; Falkenhagen, U; Naumann, G; Sokolowska-Köhler, W; Hacker, J

    1992-04-01

    A total of 36 Escherichia coli urinary tract isolates (UTI) of serotype O6, with different combinations of capsule (K) and flagellin (H) antigens, were analysed according to the outer membrane pattern (OMP), serum resistance properties, mannose-resistant hemagglutination using various types of erythrocytes, and also for the genetic presence and the expression of P-fimbriae, S fimbriae/F1C fimbriae, Type 1 fimbriae, aerobactin and hemolysin. Twenty selected strains were further analysed by pulsed field gel electrophoresis (PFGE), elaborating genomic profiles by XbaI cleavage and subsequent Southern hybridization to virulence-associated DNA probes. It could be shown that O6 UTI isolates represent a highly heterogeneous group of strains according to the occurrence and combination of these traits. Relatedness on the genetic and the phenotypic level was found for some of the strains exhibiting the same O:K:H:F serotype. DNA long-range mapping further indicated some interesting features, according to the copy number and the genomic linkage of virulence genes.

  1. Frequent acquisition of low-virulence strains of ESBL-producing Escherichia coli in travellers.

    Science.gov (United States)

    Vading, M; Kabir, M H; Kalin, M; Iversen, A; Wiklund, S; Nauclér, P; Giske, C G

    2016-12-01

    International travel is a risk factor for intestinal colonization with ESBL-producing Enterobacteriaceae (EPE). This prospective cohort study focuses on molecular features of and risk factors for travel-acquired EPE. Rectal swabs and survey data were collected from 188 Swedes travelling to four regions of high EPE prevalence. Samples were plated onto selective agars. ESBL producers were determined using phenotypic methods. Molecular characterization regarding virulence factors and phylogenetic grouping of ESBL-producing Escherichia coli was done using PCR. Isolates were also screened for the plasmid-mediated colistin resistance gene mcr-1. Among 175 pre-travel EPE-negative participants, 32% were positive upon return. No carbapenemase-producing Enterobacteriaceae were found, but one CTX-M-producing E. coli harboured mcr-1 (travel to Thailand). Most E. coli strains (43.1%) belonged to phylogroup A and were rarely associated with extraintestinal infections and a few (9.2%) expressed uropathogenicity pap genes. During 10-26 months of follow-up, no clinical infections were observed. Colonization rates varied by visited region: the Indian subcontinent, 49.2%; northern Africa, 44.0%; South-East Asia, 19.1%; and Turkey, 9.5%. Travellers' diarrhoea (OR 2.5, P = 0.04) or antimicrobial treatment during the trip (OR 5.9, P = 0.02) were both independent risk factors for EPE colonization. EPE acquired during travel have seemingly low pathogenicity, possibly indicating a low risk of clinical infection. Pre-travel advice should emphasize avoiding unnecessary antibiotic treatment during travel. © The Author 2016. Published by Oxford University Press on behalf of the British Society for Antimicrobial Chemotherapy. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

  2. Prevalence and antibiotic resistance profiles of diarrheagenic Escherichia coli strains isolated from food items in northwestern Mexico.

    Science.gov (United States)

    Canizalez-Roman, Adrian; Gonzalez-Nuñez, Edgar; Vidal, Jorge E; Flores-Villaseñor, Héctor; León-Sicairos, Nidia

    2013-06-03

    Diarrheogenic Escherichia coli (DEC) strains are an important cause of intestinal syndromes in the developing world mainly affecting children. DEC strains often infect tourists from developed countries traveling to Mexico, causing so-called "traveler diarrhea". DEC strains are typically transmitted by contaminated food and water; however, the prevalence of these strains in food items that are produced, consumed and sometimes exported in northwestern Mexico has not been evaluated. In this study, we conducted a large microbiological survey of DEC strains in 5162 food items and beverages consumed throughout Sinaloa state during 2008 and 2009. We developed a panel of eight sequential PCR reactions that detected the presence of all DEC categories, including typical or atypical variants. Thermotolerant coliforms (also known as fecal coliforms) and E. coli were detected by conventional bacteriology in 13.4% (692/5162) and 7.92% (409/5162) of food items, respectively. Among 409 E. coli isolates, 13.6% (56/409) belonged to DEC strains. Dairy products (2.8%) were the most contaminated with DEC, while DEC strains were not detected in beverages and ice samples. The pathogenic type that was most commonly isolated was EPEC (78.5%), followed by EAEC (10.7%), STEC (8.9%) and ETEC (1.7%). EHEC, DAEC and EIEC strains were not detected. Approximately 80% of EPEC and EAEC strains were classified as atypical variants; they did not adhere to a culture of HEp-2 cell. Of the isolated DEC strains, 66% showed resistance to at least one commonly prescribed antibiotic. In conclusion, the presence of DEC strains in food items and beverages available in northwestern Mexico is low and may not represent a threat for the general population or those traveling to tourist areas.

  3. MECHANISMS OF RESISTANCE TO CIPROFLOXACIN AND GENETIC DIVERSITY OF ESCHERICHIA COLI STRAINS ORIGINATING FROM URINE CULTURES PERFORMED FOR ROMANIAN ADULTS.

    Science.gov (United States)

    Cristea, Violeta Corina; Oprea, Mihaela; Neacşu, Gabriela; Gîlcă, Ramona; Popa, Mircea Ioan; Usein, Codruţa-Romaniţa

    2015-01-01

    Urinary tract infections (UTI) with Escherichia coli are among the most common infections presenting in general practice. Fluoroquinolones (FQs) are relied on for their empirical therapy but recent reports indicate a concerning increase in the percentage of FQ-resistant E. coli isolates in many countries, including Romania. Sixty E. coli strains with ciprofloxacin resistance and cephalosporin susceptibility isolated from urine specimens of non-hospitalized patients during a five-month period (October 2014 - February 2015) were further analyzed to determine the molecular basis of FQ resistance (i.e. mutations in chromosomal gyrA, gyrB, parC genes and presence of plasmid-borne qnrA, qnrB, qnrS, and aac(6'-Ib-cr genes), the phylogenetic background (i.e. phylogenetic groups A, B1, B2, C, D, E, F or clade I), O25b/ST131 status, and genetic relatedness inferred from the XbaI pulsed-field gel electrophoresis (PFGE) profiles as a measure of isolate-specific genetic composition. The PCR-based phylotyping showed that most strains were assigned to non-B2 phylogenetic groups (i.e. group A/21 strains, group B1/14 strains, group B2/10 strains, group C/8 strains, group D/3 strains, group F/4 strains). Already described chromosomal mutations associated to FQ resistance were found, the strains being double gyrA mutants (i.e. Ser83Leu, Asp87Asn) with one or two parC mutations (e.g. Ala56Thr, Ser80Ile, Glu84Gly). Seven percent of the strains harboured plasmid-borne genes qnrS1 (2 strains) and aac(6'-Ib-cr (2 strains). Based on the PCR results, 15% of the strains were members of the O25b/ST131 clone and possessed the gyrA/parC allele combination which is considered as hallmark of H30 subclone. PFGE genotyping revealed a genetically diverse population of FQ-resistant E. coli. ST131 strains displayed more homogeneous PFGE profiles than non-ST131. The ST131 cluster extended to 77.74% similarity versus 60% overall. These findings underscore the need for ongoing surveillance to capture the

  4. Engineering of Escherichia coli strains for plasmid biopharmaceutical production: scale-up challenges.

    Science.gov (United States)

    Gonçalves, Geisa A L; Prather, Kristala L J; Monteiro, Gabriel A; Prazeres, Duarte M F

    2014-05-19

    Plasmid-based vaccines and therapeutics have been making their way into the clinic in the last years. The existence of cost-effective manufacturing processes capable of delivering high amounts of high-quality plasmid DNA (pDNA) is essential to generate enough material for trials and support future commercialization. However, the development of pDNA manufacturing processes is often hampered by difficulties in predicting process scale performance of Escherichia coli cultivation on the basis of results obtained at lab scale. This paper reports on the differences observed in pDNA production when using shake flask and bench-scale bioreactor cultivation of E. coli strains MG1655ΔendAΔrecA and DH5α in complex media with 20 g/L of glucose. MG1655ΔendAΔrecA produced 5-fold more pDNA (9.8 mg/g DCW) in bioreactor than in shake flask (1.9 mg/g DCW) and DH5α produced 4-fold more pDNA (8 mg/g DCW) in bioreactor than in shake flask (2 mg/g DCW). Accumulation of acetate was also significant in shake flasks but not in bioreactors, a fact that was attributed to a lack of control of pH.

  5. Characterization of heat-shock proteins in Escherichia coli strains under thermal stress in vitro.

    Science.gov (United States)

    Urban-Chmiel, Renata; Dec, Marta; Puchalski, Andrzej; Wernicki, Andrzej

    2013-12-01

    The aim of this study was to evaluate the effect of heat stress in in vitro conditions on the induction of heat-shock protein (Hsp)70 by Escherichia coli cells, and to determine the localization of Hsps in cell fractions. The material consisted of wild strains of E. coli isolated from the digestive tract of calves, suspended in an exponential-phase culture and subjected to 41.5 °C for 2 h. Individual fractions were analysed by SDS-PAGE and two-dimensional electrophoresis. Western blotting with mouse anti-Hsp70 and anti-Hsp60 mAbs was used to identify the proteins. Electrophoretic analysis of the heat-treated cells detected Hsp70 in all three fractions, cytoplasmic, periplasmic and membrane, which was confirmed by Western blotting. The proteins obtained had diverse localizations in the pH gradient in two-dimensional electrophoresis, which may indicate changes in their conformation and physical properties leading to stabilization and protection of intracellular structures in stress conditions. The presence of these Hsps in different cell fractions indicates a very strong protective adaptation in the bacteria in unfavourable conditions, which is critical for the organism infected by them.

  6. Survival rate of campylobacter coli strains in sterile buffalo and bovine milk Tasas de sobrevida de Campylobacter coli en leche de búfalo y de bovino

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    A. TRESIERRA-AYALA

    2001-01-01

    Full Text Available The survival rate of five Campylobacter coli strains, isolated from bovine faeces (3 and from buffalo faeces (2, was studied in sterile buffalo and bovine milk kept at 4°C under aerobic conditions. All strains lost their viability substantially more rapidly in buffalo milk than in bovine milkSe determinó, en leche de bovino y de búfalo autoclavadas y mantenidas a 4ºC bajo condiciones de aerobiosis, la sobrevida de cinco cepas de Campylobacter coli aisladas de fecas de bovino (3 y de búfalo (2. Todas las cepas perdieron su viabilidad más rápidamente en leche de búfalo que en leche bovina

  7. Antimicrobial resistance, integrons and plasmid replicon typing in multiresistant clinical Escherichia coli strains from Enugu State, Nigeria.

    Science.gov (United States)

    Chah, Kennedy F; Agbo, Ifeoma C; Eze, Didacus C; Somalo, Sergio; Estepa, Vanesa; Torres, Carmen

    2010-12-01

    Eleven multiresistant Escherichia coli strains of animal and human origin were assayed for the presence of antimicrobial resistance genes, integrons and associated gene cassettes, as well as plasmid content. Ciprofloxacin-resistant strains were screened for amino acid changes in GyrA and ParC proteins. The E. coli strains were found to harbor a variety of genes including cmlA, aac (3)-II, aac (3)-IV, aadA, strA-strB, tet (A), tet (B), bla(TEM), sul1, sul2 and sul3. Four of the eight int I1-positive strains were also positive for qacE Δ1 -sul1 region and the following gene cassettes were detected: dfrA7, dfrA12 + orfF + aadA2 and bla(OXA1)+ aadA1. Five strains contained class 1 integrons lacking the qacE Δ1 -sul1 region and they showed a single type of gene cassette arrangement (estX + psp + aadA2 + cmlA + aadA1 + qacH + IS440 + sul3). The two int I2-positive strains carried the same type of gene cassette arrangement (dfrA1 + sat + aadA1). The seven ciprofloxacin-resistant E. coli strains exhibited a Ser-83-Leu substitution in GyrA protein and a Ser-80-Ile substitution in ParC protein; six of these strains presented an additional substitution in GyrA (Asp-87-Gly or Asp-87-Asn) and one strain in ParC (Glu-84-Gly). Eight different plasmid-replicon-types were detected among the 11 E. coli strains, IncF being the most frequent one detected, found in nine strains; other plasmid replicon types detected were IncX, IncI1, IncY, IncW, IncFIC, IncB/O, and IncK. Antimicrobial resistance in the E. coli strains studied was mediated by a variety of genes, some of them included in integrons, as well as by mutations gyr A and par C genes.

  8. High-virulence CMY-2- and CTX-M-2-producing avian pathogenic Escherichia coli strains isolated from commercial turkeys.

    Science.gov (United States)

    da Silva, Ketrin Cristina; Cunha, Marcos Paulo Vieira; Cerdeira, Louise; de Oliveira, Maria Gabriela Xavier; de Oliveira, Mirela Caroline Vilela; Gomes, Cleise Ribeiro; Lincopan, Nilton; Knöbl, Terezinha; Moreno, Andrea Micke

    2017-01-01

    This study reports the high-virulence phylogenetic backgrounds of CMY-2- and CTX-M-2-producing avian pathogenic Escherichia coli strains isolated from turkeys sent to slaughter and condemned by airsacculitis in Brazil. Among 300 air sac samples, seven E. coli strains produced plasmid-mediated CMY-2-type AmpC, of which three carried also the blaCTX-M-2 Extended Spectrum Beta-Lactamase encoding gene. Interestingly, the transfer of the blaCMY-2 gene was positive for three E. coli strains, being associated with the presence of IncI1 plasmids. The complete sequence of the representative pJB10 plasmid revealed that the blaCMY-2 gene was within a transposon-like element in the classical genetic environment consisting of tnpA-blaCMY-2-blc-sugE structure. This plasmid with 94-kb belonged to the sequence type (ST) 12 among IncI1 plasmids, which has been associated with the worldwide spread of blaCMY-2 among Salmonella enterica and E. coli. Furthermore, to the best of our knowledge, this is the first complete sequence of a CMY-2-encoding plasmid derived from an Escherichia coli isolated from food-producing animals in Latin America.

  9. Aerobactin and other virulence factor genes among strains of Escherichia coli causing urosepsis: association with patient characteristics.

    Science.gov (United States)

    Johnson, J R; Moseley, S L; Roberts, P L; Stamm, W E

    1988-02-01

    To assess the role of aerobactin as a virulence factor among uropathogenic Escherichia coli, we determined the prevalence, location, and phenotypic expression of aerobactin determinants among 58 E. coli strains causing bacteremic urinary tract infections. We correlated the presence of the aerobactin system with antimicrobial-agent resistance, the presence and phenotypic expression of other uropathogenic virulence factor determinants (P fimbriae, hemolysin, and type 1 fimbriae), and characteristics of patients. Colony and Southern hybridization of total and plasmid DNA with DNA probes for each virulence factor showed that aerobactin determinants were present in 78% of the strains and were plasmid associated in 21%, whereas P fimbria, hemolysin, and type 1 fimbria determinants were present in 74, 43, and 98% of the strains, respectively, and were always chromosomal. Chromosomal aerobactin, P fimbria, and hemolysin determinants occurred together on the chromosome more often in strains from patients without predisposing urological or medical conditions (P = 0.04). Strains with plasmid-encoded aerobactin lacked determinants for P fimbriae (P = 0.004) and hemolysin (P = 0.0004), were resistant to multiple antimicrobial agents (P = 0.0001), and were found only in compromised patients. Mating experiments demonstrated that some aerobactin plasmids also encoded antimicrobial-agent resistance. These findings suggest that the determinants for aerobactin, P fimbriae, and hemolysin are conserved on the chromosome of the antimicrobial-agent-susceptible uropathogenic strains of E. coli which invade noncompromised patients. In contrast, these chromosomal virulence factors are often absent from E. coli strains causing urosepsis in compromised hosts; these strains may acquire plasmid aerobactin in conjunction with antimicrobial-agent resistance genes.

  10. Multi drug resistance of campylobacter jejuni and campylobacter coli to tested antibiotics in strains originating from humans, poultry and swine

    Directory of Open Access Journals (Sweden)

    Tambur Zoran Ž.

    2010-01-01

    Full Text Available Thermophilic Campylobacter are among the most common cause of bacterial enteritis in humans. Food animals are considered one of the most important sources of Campylobacter causing infections in man. Campylobacter infection is clinically mild and resolves spontaneously. In severe or long-lasting cases, treatment with antibiotics is necessary. Resistance of Campylobacter spp. to drugs used in treatment of infection is a matter of concern. The aim of this paper is to determine presence of multi drug resistant strains of Campylobacter jejuni and Campylobacter coli isolated from animals and man. Material for testing was obtained by scraping the cecum surface from boilers, pig cecum and colon, and human feces. For isolation Campylobacter jejuni and Campylobacter coli microaerophilic conditions, temperature of 42°C and antibiotic supplement were required to inhibit the growth of other intestinal bacteria. In this research, for sensitivity testing of Campylobacter jejuni and Campylobacter coli three different methods were used: disc diffusion test, E-test, and dilution agar method. A total of 55 strains of Campylobacter jejuni and Campylobacter coli. Out of the total, 24 strains originated from man, 16 from broilers were isolated, and 15 from pigs. Multidrug resistance was determined in cases when the strains were resistant to two or more antibiotics. Applying E-test, we detected that the largest number of Campylobacter jejuni were multi drug resistant to two antibiotics (41.2%, and three antibiotics (11.8%. Applying disc diffusion method it was detected that 5.9% of Campylobacter jejuni from man was resistant to four tested antibiotics. Applying all three methods, it was detected that the largest number of Campylobacter strains was resistant to two antibiotics and three antibiotics. Applying disc diffusion method it was detected that 50% of Campylobacter coli strains from pigs were resistant to three tested antibiotics.

  11. Draft Genome Sequences of Escherichia coli Strains Isolated at Calving from the Uterus, Vagina, Vulva, and Rectoanal Junction of a Dairy Cow That Later Developed Metritis.

    Science.gov (United States)

    Jeon, Soo Jin; Cunha, Federico; Ginn, Amber; Jeong, KwangCheol Casey; Galvão, Klibs N

    2017-03-16

    Escherichia coli is involved in the pathogenicity of metritis in cows. We report here the genome sequences of E. coli strains isolated at calving from the uterus, vagina, vulva, and rectoanal junction of a dairy cow that later developed metritis. The genomic similarities will give an insight into phylogenetic relationships among strains. Copyright © 2017 Jeon et al.

  12. Adherent-invasive Escherichia coli, strain LF82 disrupts apical junctional complexes in polarized epithelia

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    Ossa Juan C

    2009-08-01

    Full Text Available Abstract Background Although bacteria are implicated in the pathogenesis of chronic inflammatory bowel diseases (IBD, mechanisms of intestinal injury and immune activation remain unclear. Identification of adherent-invasive Escherichia coli (AIEC strains in IBD patients offers an opportunity to characterize the pathogenesis of microbial-induced intestinal inflammation in IBD. Previous studies have focused on the invasive phenotype of AIEC and the ability to replicate and survive in phagocytes. However, the precise mechanisms by which these newly identified microbes penetrate the epithelial lining remain to be clarified. Therefore, the aim of this study was to delineate the effects of AIEC, strain LF82 (serotype O83:H1 on model polarized epithelial monolayers as a contributor to intestinal injury in IBD. Results Infection of T84 and Madin-Darby Canine Kidney-I polarized epithelial cell monolayers with AIEC, strain LF82 led to a reduction in transepithelial electrical resistance and increased macromolecular (10 kilodalton dextran flux. Basolateral AIEC infection resulted in more severe disruption of the epithelial barrier. Increased permeability was accompanied by a redistribution of the tight junction adaptor protein, zonula occludens-1, demonstrated by confocal microscopy and formation of gaps between cells, as shown by transmission electron microscopy. After 4 h of infection of intestine 407 cells, bacteria replicated in the cell cytoplasm and were enclosed in membrane-bound vesicles positive for the late endosomal marker, LAMP1. Conclusion These findings indicate that AIEC, strain LF82 disrupts the integrity of the polarized epithelial cell barrier. This disruption enables bacteria to penetrate into the epithelium and replicate in the host cell cytoplasm. These findings provide important links between microbes related to IBD, the intestinal epithelial cell barrier and disease pathogenesis.

  13. Investigation of sensitivity and resistance to antibiotics and chemotherapeutics in E. coli strains isolated from animals bred in intensive farming conditions

    OpenAIRE

    2005-01-01

    Being an important pathogenic and indicator microorganism, E. coli has been included in all the resistance follow up programs worldwide. We carried out sampling on 42 cattle, pig and poultry farms. After sampling the material was delivered to the laboratories, where it was exposed to standard treatment and streaking by standard methods of microbiological diagnostics with the aim to isolate and identify E. coli strains. On the selected isolated E. coli strains an investigation of its sensitivi...

  14. Persistence of Pathogenic and Non-Pathogenic Escherichia coli Strains in Various Tropical Agricultural Soils of India.

    Science.gov (United States)

    Naganandhini, S; Kennedy, Z John; Uyttendaele, M; Balachandar, D

    2015-01-01

    The persistence of Shiga-like toxin producing E. coli (STEC) strains in the agricultural soil creates serious threat to human health through fresh vegetables growing on them. However, the survival of STEC strains in Indian tropical soils is not yet understood thoroughly. Additionally how the survival of STEC strain in soil diverges with non-pathogenic and genetically modified E. coli strains is also not yet assessed. Hence in the present study, the survival pattern of STEC strain (O157-TNAU) was compared with non-pathogenic (MTCC433) and genetically modified (DH5α) strains on different tropical agricultural soils and on a vegetable growing medium, cocopeat under controlled condition. The survival pattern clearly discriminated DH5α from MTCC433 and O157-TNAU, which had shorter life (40 days) than those compared (60 days). Similarly, among the soils assessed, the red laterite and tropical latosol supported longer survival of O157-TNAU and MTCC433 as compared to wetland and black cotton soils. In cocopeat, O157 recorded significantly longer survival than other two strains. The survival data were successfully analyzed using Double-Weibull model and the modeling parameters were correlated with soil physico-chemical and biological properties using principal component analysis (PCA). The PCA of all the three strains revealed that pH, microbial biomass carbon, dehydrogenase activity and available N and P contents of the soil decided the survival of E. coli strains in those soils and cocopeat. The present research work suggests that the survival of O157 differs in tropical Indian soils due to varied physico-chemical and biological properties and the survival is much shorter than those reported in temperate soils. As the survival pattern of non-pathogenic strain, MTCC433 is similar to O157-TNAU in tropical soils, the former can be used as safe model organism for open field studies.

  15. Persistence of Pathogenic and Non-Pathogenic Escherichia coli Strains in Various Tropical Agricultural Soils of India.

    Directory of Open Access Journals (Sweden)

    S Naganandhini

    Full Text Available The persistence of Shiga-like toxin producing E. coli (STEC strains in the agricultural soil creates serious threat to human health through fresh vegetables growing on them. However, the survival of STEC strains in Indian tropical soils is not yet understood thoroughly. Additionally how the survival of STEC strain in soil diverges with non-pathogenic and genetically modified E. coli strains is also not yet assessed. Hence in the present study, the survival pattern of STEC strain (O157-TNAU was compared with non-pathogenic (MTCC433 and genetically modified (DH5α strains on different tropical agricultural soils and on a vegetable growing medium, cocopeat under controlled condition. The survival pattern clearly discriminated DH5α from MTCC433 and O157-TNAU, which had shorter life (40 days than those compared (60 days. Similarly, among the soils assessed, the red laterite and tropical latosol supported longer survival of O157-TNAU and MTCC433 as compared to wetland and black cotton soils. In cocopeat, O157 recorded significantly longer survival than other two strains. The survival data were successfully analyzed using Double-Weibull model and the modeling parameters were correlated with soil physico-chemical and biological properties using principal component analysis (PCA. The PCA of all the three strains revealed that pH, microbial biomass carbon, dehydrogenase activity and available N and P contents of the soil decided the survival of E. coli strains in those soils and cocopeat. The present research work suggests that the survival of O157 differs in tropical Indian soils due to varied physico-chemical and biological properties and the survival is much shorter than those reported in temperate soils. As the survival pattern of non-pathogenic strain, MTCC433 is similar to O157-TNAU in tropical soils, the former can be used as safe model organism for open field studies.

  16. Advanced treatment of urban wastewater by UV radiation: Effect on antibiotics and antibiotic-resistant E. coli strains.

    Science.gov (United States)

    Rizzo, Luigi; Fiorentino, Antonino; Anselmo, Antonella

    2013-06-01

    Urban wastewater treatment plant (UWWTP) effluents are among the possible sources of antibiotics and antibiotic-resistant bacteria (ARB) spread into the environment. In this work, the effect of UV radiation on antibiotic-resistant Escherichia coli (E. coli) strains was compared with that of chlorination process. Under the investigated conditions, UV disinfection process resulted in a total inactivation after 60min of irradiation (1.25×10(4)μWscm(-2)) compared to 120min chlorine contact time (initial chlorine dose of 2mgL(-1)). Moreover, no change in E. coli strains' resistance to amoxicillin (AMX) (minimum inhibiting concentration (MIC)>256mgL(-1)) and sulfamethoxazole (SMZ) (MIC>1024mgL(-1)) could be observed after UV treatment, while the treatment affected resistance of the lower resistance strain to ciprofloxacin (CPX) (MIC decreased by 33% and 50% after 60 and 120min, respectively). Contrarily, chlorination process did not affect antibiotic resistance of the investigated E. coli strains. Finally, the effect of UV radiation on the mixture of three antibiotics was also investigated and photodegradation data fit quite well pseudo first order kinetic models with t1/2 values of 14, 20 and 25min for CPX, AMX and SMZ, respectively. According to these results, conventional disinfection processes may not be effective in the inactivation of ARB, and the simultaneous release of ARB and antibiotics at sub-lethal concentrations into UWWTP effluent may promote the development of resistance among bacteria in receiving water.

  17. Draft Genome Sequence of Campylobacter coli Strain IPSID-1 Isolated from a Patient with Immunoproliferative Small Intestinal Disease.

    Science.gov (United States)

    Criscuolo, Alexis; de la Blanchardière, Arnaud; Coeuret, Solène; Passet, Virginie; Saguet-Rysanek, Virginie; Vergnaud, Michel; Verdon, Renaud; Leclercq, Alexandre; Lecuit, Marc; Brisse, Sylvain

    2014-03-13

    The genome sequence and annotation of Campylobacter coli strain IPSID-1 are reported here. This bacterial isolate is the first to be cultured from a patient with immunoproliferative small intestinal disease (IPSID). The draft genome sequence is 1.683 Mb long, comprises 64 contigs, and has 31.26% G+C content.

  18. Isolation and characterization of bacteriophages as potential agents against Shiga toxin – producing Escherichia coli (STEC) strains

    Science.gov (United States)

    Shiga – toxin producing Escherichia coli (STEC) is a significant group of foodborne pathogens that can cause mild diarrhea to serious human illnesses. The gastrointestinal tracts of cattle and other ruminants are the primary reservoirs of STEC strains and may co-harbor bacteriophages as part of its ...

  19. Draft Genome Sequence of Five Shiga Toxin-Producing Escherichia coli Strains Isolated from Wild Deer in Japan

    Science.gov (United States)

    Ikeda, Tetsuya; Yamamoto, Shiori; Kabeya, Hidenori; Sugiyama, Hiromu; Takai, Shinji

    2017-01-01

    ABSTRACT Shiga toxin-producing Escherichia coli (STEC) is one of the major foodborne pathogens. Having observed the wide distribution of this pathogen in wild deer, we report here the draft genome sequence of five STEC strains isolated from wild deer (Cervus nippon yesoensis) in Hokkaido, Japan. PMID:28254967

  20. Rapid MALDI-TOF mass spectrometry strain typing during a large outbreak of Shiga-Toxigenic Escherichia coli.

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    Martin Christner

    Full Text Available BACKGROUND: In 2011 northern Germany experienced a large outbreak of Shiga-Toxigenic Escherichia coli O104:H4. The large amount of samples sent to microbiology laboratories for epidemiological assessment highlighted the importance of fast and inexpensive typing procedures. We have therefore evaluated the applicability of a MALDI-TOF mass spectrometry based strategy for outbreak strain identification. METHODS: Specific peaks in the outbreak strain's spectrum were identified by comparative analysis of archived pre-outbreak spectra that had been acquired for routine species-level identification. Proteins underlying these discriminatory peaks were identified by liquid chromatography tandem mass spectrometry and validated against publicly available databases. The resulting typing scheme was evaluated against PCR genotyping with 294 E. coli isolates from clinical samples collected during the outbreak. RESULTS: Comparative spectrum analysis revealed two characteristic peaks at m/z 6711 and m/z 10883. The underlying proteins were found to be of low prevalence among genome sequenced E. coli strains. Marker peak detection correctly classified 292 of 293 study isolates, including all 104 outbreak isolates. CONCLUSIONS: MALDI-TOF mass spectrometry allowed for reliable outbreak strain identification during a large outbreak of Shiga-Toxigenic E. coli. The applied typing strategy could probably be adapted to other typing tasks and might facilitate epidemiological surveys as part of the routine pathogen identification workflow.

  1. Simultaneous direct detection of Shiga-toxin producing Escherichia coli (STEC) strains by gold nanoparticle optical sensing

    Science.gov (United States)

    Shiga-toxin producing Escherichia coli (STEC) strains (“Big Six” – O26, O45, O103, O111, O121, O145, and O157) represent significant groups of pathogens responsible for foodborne diseases. The objective of this study was to develop a colorimetric optical sensing assay that can simultaneously detect ...

  2. Effect of solar radiation on multidrug resistant E. coli strains and antibiotic mixture photodegradation in wastewater polluted stream

    Energy Technology Data Exchange (ETDEWEB)

    Rizzo, L., E-mail: l.rizzo@unisa.it [Department of Civil Engineering, University of Salerno, via Ponte don Melillo, 1-84084 Fisciano (Italy); Fiorentino, A. [Department of Civil Engineering, University of Salerno, via Ponte don Melillo, 1-84084 Fisciano (Italy); Anselmo, A. [Pluriacque, via Alento, 84060 Prignano Cilento (Italy)

    2012-06-15

    The effect of solar radiation on the inactivation of multidrug resistant Escherichia coli (MDR) strains selected from an urban wastewater treatment plant (UWWTP) effluent and the change of their resistance to a mixture of three antibiotics (evaluated in terms of minimum inhibit concentration (MIC)) in wastewater polluted stream were investigated. The solar photodegradation of the mixture of the three target antibiotics (amoxicillin (AMX), ciprofloxacin (CPX), and sulfamethoxazole (SMZ)) was also evaluated. Additionally, since UWWTP effluents are possible sources of antibiotics and antibiotic resistant bacteria, the disinfection by conventional chlorination process of the UWWTP effluent inoculated with MDR strains was investigated too. Solar radiation poorly affected the inactivation of the two selected antibiotic resistant E. coli strains (40 and 60% after 180 min irradiation). Moreover, solar radiation did not affect strain resistance to AMX (MIC > 256 {mu}g/mL) and SMZ (MIC > 1024 {mu}g/mL), but affected resistance of the lower resistance strain to CPX (MIC decreased by 33% but only after 180 min of irradiation). Chlorination of wastewater sample strongly decreased the number of the two selected antibiotic resistant E. coli strains (99.667 and 99.999%), after 60 min of contact time at 2.0 mg/L initial chlorine concentration, but the resistance of survived colonies to antibiotics was unchanged. Finally, the solar photodegradation rate of the antibiotic mixture (1 mg/L initial concentration respectively) resulted in the following order (half-life time): CPX (t{sub 1/2} = 24 min) < AMX (t{sub 1/2} = 99 min) < SMZ (t{sub 1/2} = 577 min). Accordingly, the risk of the development of resistance to SMZ in surface water is significantly higher compared to CPX and AMX. - Highlights: Black-Right-Pointing-Pointer Solar radiation did not affect E. coli strain resistance to AMX and SMZ. Black-Right-Pointing-Pointer Solar radiation affected the resistance of one E. coli strain

  3. Distribution of strain type and antimicrobial susceptibility of Escherichia coli isolates causing meningitis in a large urban setting in Brazil.

    Science.gov (United States)

    Berman, Hillary; Barberino, Maria Goreth; Moreira, Edson Duarte; Riley, Lee; Reis, Joice N

    2014-05-01

    The clinical management of meningitis caused by Escherichia coli is greatly complicated when the organism becomes resistant to broad-spectrum antibiotics. We sought to characterize the antimicrobial susceptibilities, sequence types (ST), and presence of known drug resistance genes of E. coli isolates that caused meningitis between 1996 and 2011 in Salvador, Brazil. We then compared these findings to those for E. coli isolates from community-acquired urinary tract infections (UTI) that occurred during the same time period and in the same city. We found that 19% of E. coli isolates from cases of meningitis and less than 1% of isolates from UTI were resistant to third-generation cephalosporins. The sequence types of E. coli isolates from cases of meningitis included ST131, ST69, ST405, and ST62, which were also found among isolates from UTI. Additionally, among the E. coli isolates that were resistant to third-generation cephalosporins, we found genes that encode the extended-spectrum beta-lactamases CTX-M-2, CTX-M-14, and CTX-M-15. These observations demonstrate that compared to E. coli strains isolated from cases of community-acquired UTI, those isolated from cases of meningitis are more resistant to third-generation cephalosporins, even though the same sequence types are shared between the two forms of extraintestinal infections.

  4. The nature of laboratory domestication changes in freshly isolated Escherichia coli strains.

    Science.gov (United States)

    Eydallin, Gustavo; Ryall, Ben; Maharjan, Ram; Ferenci, Thomas

    2014-03-01

    Adaptation of environmental bacteria to laboratory conditions can lead to modification of important traits, what we term domestication. Little is known about the rapidity and reproducibility of domestication changes, the uniformity of these changes within a species or how diverse these are in a single culture. Here, we analysed phenotypic changes in nutrient-rich liquid media or on agar of four Escherichia coli strains newly isolated through minimal steps from different sources. The laboratory-cultured populations showed changes in metabolism, morphotype, fitness and in some phenotypes associated with the sigma factor RpoS. Domestication events and phenotypic diversity started to emerge within 2-3 days in replicate subcultures of the same ancestor. In some strains, increased amino acid usage and higher fitness under nutrient limitation resembled those in mutants with the GASP (growth advantage in stationary phase) phenotype. The domestication changes are not uniform across a species or even within a single domesticated population. However, some parallelism in adaptation within repeat cultures was observed. Differences in the laboratory environment also determine domestication effects, which differ between liquid and solid media or with extended stationary phase. Important lessons for the handling and storage of organisms can be based on these studies.

  5. Characterization of spontaneous mutation in the oxyR strain of Escherichia coli.

    Science.gov (United States)

    Yamamura, E; Nunoshiba, T; Kawata, M; Yamamoto, K

    2000-12-20

    Escherichia coli K-12 strain EY5, deficient in oxyR, was constructed to assess the role of oxyR and oxyR-regulated regulon in spontaneous mutagenesis. Mutagenesis was monitored by selecting two forward mutations of colicin B-sensitive to resistance and valine-sensitive to resistance, one base substitution mutation of rifampicin-sensitive to resistance and one reversion of argE3 his-4 to Arg(+) His(+). Deficiency of oxyR did not lead to the enhancement of spontaneous mutation frequencies of the four markers tested. By DNA sequence analysis, we determined 49 colicin B-resistant mutants derived from EY5 and found that 37% were base substitutions, 29% IS element insertions, 20% deletions, and 14% single base frameshifts. Among the base substitutions, G:C-->T:A transversions predominated followed by G:C-->A:T transitions and A:T-->T:A transversions. These spectra were essentially the same as those from oxyR(+) strains. The results indicate that oxyR and oxyR-regulated genes do not play a significant role in the defense against spontaneous mutagenesis.

  6. A pathoadaptive deletion in an enteroaggregative Escherichia coli outbreak strain enhances virulence in a Caenorhabditis elegans model.

    Science.gov (United States)

    Hwang, Jennifer; Mattei, Lisa M; VanArendonk, Laura G; Meneely, Philip M; Okeke, Iruka N

    2010-09-01

    Enteroaggregative Escherichia coli (EAEC) strains are important diarrheal pathogens. EAEC strains are defined by their characteristic stacked-brick pattern of adherence to epithelial cells but show heterogeneous virulence and have different combinations of adhesin and toxin genes. Pathoadaptive deletions in the lysine decarboxylase (cad) genes have been noted among hypervirulent E. coli subtypes of Shigella and enterohemorrhagic E. coli. To test the hypothesis that cad deletions might account for heterogeneity in EAEC virulence, we developed a Caenorhabditis elegans pathogenesis model. Well-characterized EAEC strains were shown to colonize and kill C. elegans, and differences in virulence could be measured quantitatively. Of 49 EAEC strains screened for lysine decarboxylase activity, 3 tested negative. Most notable is isolate 101-1, which was recovered in Japan, from the largest documented EAEC outbreak. EAEC strain 101-1 was unable to decarboxylate lysine in vitro due to deletions in cadA and cadC, which, respectively, encode lysine decarboxylase and a transcriptional activator of the cadAB genes. Strain 101-1 was significantly more lethal to C. elegans than control strain OP50. Lethality was attenuated when the lysine decarboxylase defect was complemented from a multicopy plasmid and in single copy. In addition, restoring lysine decarboxylase function produced derivatives of 101-1 deficient in aggregative adherence to cultured human epithelial cells. Lysine decarboxylase inactivation is pathoadapative in an important EAEC outbreak strain, and deletion of cad genes could produce hypervirulent EAEC lineages in the future. These results suggest that loss, as well as gain, of genetic material can account for heterogeneous virulence among EAEC strains.

  7. Occurrence of Hybrid Escherichia coli Strains Carrying Shiga Toxin and Heat-Stable Toxin in Livestock of Bangladesh

    Science.gov (United States)

    Johura, Fatema-Tuz; Parveen, Rozina; Islam, Atiqul; Sadique, Abdus; Rahim, Md Niaz; Monira, Shirajum; Khan, Anisur R.; Ahsan, Sunjukta; Ohnishi, Makoto; Watanabe, Haruo; Chakraborty, Subhra; George, Christine M.; Cravioto, Alejandro; Navarro, Armando; Hasan, Badrul; Alam, Munirul

    2017-01-01

    Shiga toxin-producing Escherichia coli (STEC) and enterotoxigenic E. coli (ETEC) are important causes of diarrhea in humans and animals worldwide. Although ruminant animals are the main source of STEC, diarrhea due to this pathotype is very low in Bangladesh where ETEC remains the predominant group associated with childhood diarrhea. In the present study, E. coli strains (n = 35) isolated from Bangladesh livestock (goats, sheep, and cattle) and poultry (chicken and ducks) were analyzed for the presence of major virulence factors, such as Shiga toxins (STX-1 and STX-2), heat-labile toxin, and heat-stable toxins (STa and STb). Multiplex polymerase chain reaction results revealed 23 (66%) E. coli strains to be virulent possessing either sta (n = 5), stx (stx1, n = 8; stx2, n = 2), or both (n = 8) genes in varying combinations. Thirty-four percent (8/23) of strains from livestock were hybrid type that carried both stx (either stx1 or stx2) and ETEC-specific enterotoxin gene sta. Serotyping results revealed that the ETEC strains belonged to five serotypes, namely O36:H5, O174:H−, O152:H8, O109:H51, and O8:H21, while the STEC-producing strains belonged to serotypes O76:H19 (n = 3), O43:H2 (n = 2), O87:H16 (n = 2), OR:H2 (n = 1), O110:H16 (n = 1), and O152:H8 (n = 1). The STEC–ETEC hybrid strains belonged to serotypes O76:H19 (n = 3), O43:H2 (n = 2), O87:H16, OR:H2, and O152:H8. Forty percent (2/5) of the ETEC and 20% (2/10) of the STEC strains were multidrug resistant with the highest drug resistance (50%) being found in the hybrid strains. Molecular fingerprinting determined by pulsed-field gel electrophoresis and cluster analyses by dendrogram revealed that, genetically, STEC–ETEC hybrid strains were highly heterogeneous. Multidrug-resistant E. coli STEC–ETEC hybrid strains in domesticated animals pose a public health threat for humans in Bangladesh. PMID:28119905

  8. Shiga toxigenic Escherichia coli show strain dependent reductions under dry-fermented sausage production and post-processing conditions.

    Science.gov (United States)

    Rode, Tone Mari; Holck, Askild; Axelsson, Lars; Høy, Martin; Heir, Even

    2012-04-16

    Dry-fermented sausages (DFS) are considered possible risk products regarding Shiga toxigenic Escherichia coli (STEC). We have compared the reduction of 11 E. coli isolates of various serogroups in salami during the sausage production process and during post-process measures including storage, heating and freezing. The 11 E. coli isolates, mainly STEC, included enterohaemorrhagic E. coli (EHEC) outbreak strains linked to DFS along with apathogenic E. coli. During sausage production, there was a statistically significant difference in reduction between the E. coli strains ranging from 1.3 to 2.4 log₁₀ (ppost-process heat treatment of 43 °C for 24 h, a total reduction of more than 5 log₁₀ was obtained for all E. coli isolates. Freezing and thawing of DFS with subsequent storage for 1 month at 16 or 20 °C generally contributed to large E. coli reductions with the latter conditions giving an average additional 3.9 log₁₀ reduction, with a range from 3.4 to 4.4 log₁₀. The combination of freezing and 1 month of storage gave higher reductions compared with storage for 2 months for all examined temperatures. No systematic differences in survival of E. coli of different serogroups were detected for the different post-process measures. The reductions were also similar to those of apathogenic control isolates. Isolates showing higher survival during the ripening process did not have a lower reduction when exposed to post-process stress like storage, heating and freezing. The ability of the isolates to survive in salami was also compared with their survival at equivalent conditions in a tryptic soy broth (TSB) model. There was a low and not significant correlation (p>0.1) between the reductions of E. coli in salami and in the TSB broth model. Results based on broth models and/or single or surrogate strains must therefore be interpreted with caution. The EHEC reducing post-processing measures tested can easily be implemented in DFS production with marginal

  9. Alignment-free design of highly discriminatory diagnostic primer sets for Escherichia coli O104:H4 outbreak strains.

    Science.gov (United States)

    Pritchard, Leighton; Holden, Nicola J; Bielaszewska, Martina; Karch, Helge; Toth, Ian K

    2012-01-01

    An Escherichia coli O104:H4 outbreak in Germany in summer 2011 caused 53 deaths, over 4000 individual infections across Europe, and considerable economic, social and political impact. This outbreak was the first in a position to exploit rapid, benchtop high-throughput sequencing (HTS) technologies and crowdsourced data analysis early in its investigation, establishing a new paradigm for rapid response to disease threats. We describe a novel strategy for design of diagnostic PCR primers that exploited this rapid draft bacterial genome sequencing to distinguish between E. coli O104:H4 outbreak isolates and other pathogenic E. coli isolates, including the historical hæmolytic uræmic syndrome (HUSEC) E. coli HUSEC041 O104:H4 strain, which possesses the same serotype as the outbreak isolates. Primers were designed using a novel alignment-free strategy against eleven draft whole genome assemblies of E. coli O104:H4 German outbreak isolates from the E. coli O104:H4 Genome Analysis Crowd-Sourcing Consortium website, and a negative sequence set containing 69 E. coli chromosome and plasmid sequences from public databases. Validation in vitro against 21 'positive' E. coli O104:H4 outbreak and 32 'negative' non-outbreak EHEC isolates indicated that individual primer sets exhibited 100% sensitivity for outbreak isolates, with false positive rates of between 9% and 22%. A minimal combination of two primers discriminated between outbreak and non-outbreak E. coli isolates with 100% sensitivity and 100% specificity. Draft genomes of isolates of disease outbreak bacteria enable high throughput primer design and enhanced diagnostic performance in comparison to traditional molecular assays. Future outbreak investigations will be able to harness HTS rapidly to generate draft genome sequences and diagnostic primer sets, greatly facilitating epidemiology and clinical diagnostics. We expect that high throughput primer design strategies will enable faster, more precise responses to

  10. Hydrogen-producing Escherichia coli strains overexpressing lactose permease: FT-IR analysis of the lactose-induced stress.

    Science.gov (United States)

    Grube, Mara; Dimanta, Ilze; Gavare, Marita; Strazdina, Inese; Liepins, Janis; Juhna, Talis; Kalnenieks, Uldis

    2014-01-01

    The lactose permease gene (lacY) was overexpressed in the septuple knockout mutant of Escherichia coli, previously engineered for hydrogen production from glucose. It was expected that raising the lactose transporter activity would elevate the intracellular lactose concentration, inactivate the lactose repressor, induce the lactose operon, and as a result stimulate overall lactose consumption and conversion. However, overexpression of the lactose transporter caused a considerable growth delay in the recombinant strain on lactose, resembling to some extent the "lactose killing" phenomenon. Therefore, the recombinant strain was subjected to selection on lactose-containing media. Selection on plates with 3% lactose yielded a strain with a decreased content of the recombinant plasmid but with an improved ability to grow and produce hydrogen on lactose. Macromolecular analysis of its biomass by means of Fourier transform-infrared spectroscopy demonstrated that increase of the cellular polysaccharide content might contribute to the adaptation of E. coli to lactose stress.

  11. [Sensitivity assessment of thyme and lavender essential oils against clinical strains of Escherichia coli for their resistance].

    Science.gov (United States)

    Sienkiewicz, Monika; Kalemba, Danuta; Wasiela, Małgorzata

    2011-01-01

    Strong antiseptic activity of plant essential oils and extracts has been known for a long time. The antibacterial activity of thyme and lavender essential oils were tested against 30 clinical bacterial strains of Escherichia coli from patients with different clinical conditions. The agar diffusion method was used for microbial growth inhibition at various concentrations of the oils from Thymus vulgaris and Lavandula angustifolia. Susceptibility testing to antibiotics and chemotherapeutics was carried out using disc-diffusion method. The results of experiments showed that the both oils, from T. vulgaris and L. angustifolia were active against all of the clinical strains, but thyme oil demonstrated the highest activity. Thyme and lavender essential oils were active against multi drug resistant clinical strains of Escherichia coli genera. The results of experiments justify a study related to activity other essential oils against different genus of bacteria.

  12. Rapid emergence of high-level tigecycline resistance in Escherichia coli strains harbouring blaNDM-5 in vivo.

    Science.gov (United States)

    Li, Xi; Mu, Xinli; Yang, Yunxing; Hua, Xiaoting; Yang, Qing; Wang, Nanfei; Du, Xiaoxing; Ruan, Zhi; Shen, Xiaoqiang; Yu, Yunsong

    2016-04-01

    Tigecycline (TIG) resistance is a growing concern because this antibiotic is regarded as one of the last resorts to treat infections caused by multidrug-resistant and extensively drug-resistant (XDR) bacteria. Information regarding TIG-resistant Escherichia coli isolates is scarce. In this study, we report the emergence of high-level TIG resistance in a longitudinal series of XDR E. coli isolates collected during TIG treatment. Whole-genome sequencing was performed for six E. coli strains harbouring bla(NDM-5) and genomic comparison revealed two amino acid substitutions. Mutation in rpsJ could be a significant factor conferring TIG resistance in these isolates. The fitness cost of TIG resistance in resistant strains was evaluated by determining the relative growth rate, indicating that TIG resistance reduced fitness by ca. 7%. This study is the first report to demonstrate high-level TIG resistance in E. coli in vivo. In addition, we report the first treatment-emergent minimum inhibitory concentration (MIC) development of TIG from 1mg/L to 64 mg/L in E. coli. Clinicians should be aware of the risk of an increase in the MIC of TIG under therapy.

  13. Molecular characterization of enterohemorrhagic Escherichia coli hemolysin gene (EHEC-hlyA)-harboring isolates from cattle reveals a diverse origin and hybrid diarrheagenic strains.

    Science.gov (United States)

    Askari Badouei, Mahdi; Morabito, Stefano; Najafifar, Arash; Mazandarani, Emad

    2016-04-01

    In the present study we investigated the occurrence of Escherichia coli strains harboring the gene encoding enterohemorrhagic E. coli hemolysin (EHEC-HlyA) in cattle and the association of this gene with various diarrheagenic E. coli (DEC) pathotypes. First, the bovine E. coli isolates were screened for EHEC-hlyA gene by PCR, and then they were characterized for the phylogenetic groups and the presence of the major virulence genes of different DEC pathotypes. In total, 25 virulence gene profiles were observed in 54 EHEC-hlyA+ isolates that reflect a considerable heterogeneity. The EHEC-hlyA+ strains were mostly associated with EHEC (72%), while only 7.4% were enteropathogenic E. coli (EPEC). We also showed the presence of estA gene of enterotoxigenic E. coli (ETEC) in 6 isolates (11.1%). Interestingly, two of the estA+ strains showed hybrid pathotypes with one carrying eae/estA (EPEC/ETEC), and the other one stx2/astA/estA (EHEC/ETEC). None of the isolates were related to enteroaggregative E. coli (EAggEC), enteroinvasive E. coli (EIEC), and necrotoxigenic E. coli (NTEC). The EHEC-plasmid encoded genes occurred in seven different combinations with EHEC-hlyA/saa/subA/espP being the most prevalent (46.3%). All stx-/eae+ strains carried O island 57 (OI-57) molecular marker(s) that may indicate these to be the progenitors of EHEC or strains losing stx. The most prevalent phylogroup was B1 (61.1%), but the most heterogeneous strains including the hybrid strains belonged to A phylogroup. Overall, our results indicate that cattle EHEC-hlyA encoding E. coli isolates consist of diverse diarrheagenic strains with the possible existence of hybrid pathotypes. Future studies are required to clarify the evolutionary aspects and clinical significance of these strains in humans and domestic animals.

  14. Colicins U and Y inhibit growth of Escherichia coli strains via recognition of conserved OmpA extracellular loop 1.

    Science.gov (United States)

    Bosák, Juraj; Micenková, Lenka; Doležalová, Magda; Šmajs, David

    2016-11-01

    Interactions of colicins U and Y with the OmpA (Outer membrane protein A) receptor molecule were studied using site-directed mutagenesis and colicin binding assay. A systematic mutagenesis of the colicin-susceptible OmpA sequence from Escherichia coli (OmpAEC) to the colicin-resistant OmpA sequence from Serratia marcescens (OmpASM) was performed in regions corresponding to extracellular OmpA loops 1-4. Susceptibility to colicins U and Y was significantly affected by the OmpA mutation in loop 1. As with functional analysis, a decrease in binding capacity of His-tagged colicin U was found for recombinant OmpA with a mutated segment in loop 1 compared to control OmpAEC. To verify the importance of the identified amino acid residues in OmpA loop 1, we introduced loop 1 from OmpAEC into OmpASM, which resulted in the substantial increase of susceptibility to colicins U and Y. In addition, colicins U and Y were tested against a panel of 118 bacteriocin non-producing strains of four Escherichia species, including E. coli (39 strains), E. fergusonii (10 strains), E. hermannii (42 strains), and E. vulneris (27 strains). A majority (82%) of E. coli strains was susceptible to colicins U and Y. Interestingly, colicins U and Y also inhibited all of the 30 tested multidrug-resistant E. coli O25b-ST131 isolates. These findings, together with the fact that OmpA loop 1 is important for bacterial virulence and is evolutionary conserved, offer the potential of using colicins U and Y as specific anti-OmpA loop 1 directed antibacterial proteins.

  15. ISOLASI STREPTOMYCES SPP. PADA KAWASAN HUTAN PROVINSI BALI SERTA UJI DAYA HAMBATNYA TERHADAP LIMA STRAIN DIARRHEAGENIC ESCHERICHIA COLI

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    I WAYAN EKA DHARMAWAN

    2014-04-01

    Full Text Available An exploration study of natural resources soil bacteria antibiotic-producer, Streptomyces spp. was done in two steps. The first step was isolation of Streptomyces and the second involved testing their inhibition activities against five strains diarrheagenic Escherichia coli. Soil samples were collected from ten forest areas in Bali. As many as 55 isolates were collected with various macroscopic dan microscopic characters. Most isolates (eight Streptomyces isolates were collected from forest area in Penulisan, Kintamani (RTK. 20. The diversities of isolates are influenced by environment condition. All Streptomyces isolated were tested against five strains diarrheagenic Escherichia coli to check antibiotic activity for inhibit growth of E. coli. Streptomycine was used as a control. The result showed that the largest inhibition zones of Streptomyces against E. coli strains EHEC, ETEC, EIEC, EPEC and DAEC were produced by Streptomyces PK5 (48,67 ± 0,58 mm, Streptomyces GAA4 (29,00 ± 2,00 mm, Streptomyces GBK3 (42,67 ± 2,08 mm, Streptomyces SkBB5 (29,00 ± 2,65 mm and Streptomyces GM3 (33,67 ± 3,21 mm respectively.

  16. Fluoroquinolone-resistance mechanisms and phylogenetic background of clinical Escherichia coli strains isolated in south-east Poland.

    Science.gov (United States)

    Korona-Glowniak, Izabela; Skrzypek, Kinga; Siwiec, Radosław; Wrobel, Andrzej; Malm, Anna

    2016-07-01

    Fluorochinolones are a class of broad-spectrum antimicrobials in the treatment of several infections, including those caused by Escherichia coli. Due to the increasing resistance of bacteria to antimicrobials, an understanding of fluoroquinolone resistance is important for infection control. The aim of this study was to determine susceptibility of clinical E. coli strains to fluoroquinolones and characterize their mechanisms of quinolone resistance. Totally, 79 non-duplicate clinical E. coli isolates included in this study were mainly from skin lesion -36 (45.6%) isolates; 54 (68.4%) isolates were assigned to phylogenetic B2 group. Resistance to ciprofloxacin was found in 20 isolates. In the quinolone resistance-determining region (QRDR) region of gyrA and parC, 4 types of point mutations were detected. Mutations in parC gene were found in all strains with gyrA mutations. Predominance of double mutation in codon 83 and 87 of gyrA (90%) and in codon 80 of parC (90%) was found. Moreover, plasmid-mediated quinolone resistance (PMRQ) determinants (qnrA or qnrB and/or aac(6')-Ib-cr) were present in 5 (25%) out of 20 fluoroquinolone-resistant isolates. Resistance to fluoroquinolones in all of the tested clinical E. coli isolates correlated with point mutations in both gyrA and parC. The majority of fluoroquinolone-resistant strains belonged to D and B2 phylogenetic groups.

  17. Rapid MALDI-TOF Mass Spectrometry Strain Typing during a Large Outbreak of Shiga-Toxigenic Escherichia coli

    Science.gov (United States)

    Christner, Martin; Trusch, Maria; Rohde, Holger; Kwiatkowski, Marcel; Schlüter, Hartmut; Wolters, Manuel; Aepfelbacher, Martin; Hentschke, Moritz

    2014-01-01

    Background In 2011 northern Germany experienced a large outbreak of Shiga-Toxigenic Escherichia coli O104:H4. The large amount of samples sent to microbiology laboratories for epidemiological assessment highlighted the importance of fast and inexpensive typing procedures. We have therefore evaluated the applicability of a MALDI-TOF mass spectrometry based strategy for outbreak strain identification. Methods Specific peaks in the outbreak strain’s spectrum were identified by comparative analysis of archived pre-outbreak spectra that had been acquired for routine species-level identification. Proteins underlying these discriminatory peaks were identified by liquid chromatography tandem mass spectrometry and validated against publicly available databases. The resulting typing scheme was evaluated against PCR genotyping with 294 E. coli isolates from clinical samples collected during the outbreak. Results Comparative spectrum analysis revealed two characteristic peaks at m/z 6711 and m/z 10883. The underlying proteins were found to be of low prevalence among genome sequenced E. coli strains. Marker peak detection correctly classified 292 of 293 study isolates, including all 104 outbreak isolates. Conclusions MALDI-TOF mass spectrometry allowed for reliable outbreak strain identification during a large outbreak of Shiga-Toxigenic E. coli. The applied typing strategy could probably be adapted to other typing tasks and might facilitate epidemiological surveys as part of the routine pathogen identification workflow. PMID:25003758

  18. Comparison of the immune responses associated with experimental bovine mastitis caused by different strains of Escherichia coli.

    Science.gov (United States)

    Blum, Shlomo E; Heller, Elimelech D; Jacoby, Shamay; Krifucks, Oleg; Leitner, Gabriel

    2017-05-01

    We studied the mammary immune response to different mammary pathogenic Escherichia coli (MPEC) strains in cows, hypothesising that the dynamics of response would differ. E. coli is a major aetiologic agent of acute clinical bovine mastitis of various degrees of severity with specific strains being associated with persistent infections. We compared challenge with three distinct pathogenic MPEC strains (VL2874, VL2732 and P4), isolated from different forms of mastitis (per-acute, persistent and acute, respectively). A secondary objective was to verify the lack of mammary pathogenicity of an environmental isolate (K71) that is used for comparison against MPEC in genomic and phenotypic studies. Twelve cows were challenged by intra-mammary infusion with one of the strains. Cellular and chemokine responses and bacterial culture follow-up were performed for 35 d. All cows challenged by any of the MPEC strains developed clinical mastitis. Differences were found in the intensity and duration of response, in somatic cell count, secreted cytokines (TNF-α, IL-6 and IL-17) and levels of milk leucocyte membrane Toll-like receptor 4 (TLR4). A sharp decrease of TLR4 on leucocytes was observed concomitantly to peak bacterial counts in milk. Intra-mammary infusion of strain K71 did not elicit inflammation and bacteria were not recovered from milk. Results suggest some differences in the mammary immune response to distinct MPEC strains that could be correlated to their previously observed pathogenic traits. This is also the first report of an E. coli strain that is non-pathogenic to the bovine mammary gland.

  19. Isolation and molecular characterization of multidrug-resistant strains of Escherichia coli and Salmonella from retail chicken meat in Japan.

    Science.gov (United States)

    Ahmed, Ashraf M; Shimabukuro, Hirofumi; Shimamoto, Tadashi

    2009-09-01

    Sixty-nine Escherichia coli and 10 Salmonella isolates, recovered from retail chicken meat in Hiroshima prefecture, Japan, were assayed for antimicrobial susceptibility, the presence of integrons and antimicrobial resistance genes. Twenty-eight out of 69 (40.6%) of E. coli and all 10 Salmonella isolates were exhibited multidrug resistance phenotypes. The most commonly reported resistance phenotypes were against ampicillin, streptomycin, spectinomycin, kanamycin, tetracycline, and trimethoprim/sulfamethoxazole. PCR screening for integrons showed that 8 (11.6%) of the E. coli isolates were positive for the class 1 integrons and 1 isolate (1.4%) was positive for the class 2 integrons. Among the 10 Salmonella isolates, 9 were positive for class 1 integrons and none was positive for class 2 integrons. The identified antibiotic resistance gene cassettes within the class 1 integrons were dfrA1, dfrA7, aadA1, aadB, and catB3, while dfrA1, sat2, and aadA1 were identified within class 2 integron. The beta-lactamase resistance gene bla(TEM-1) was identified in 12 (17.3%) of E. coli isolates and in only one of the Salmonella isolates. The bla(CMY-2) gene, encoding AmpC beta-lactamase, was detected in 16 (23.2%) of the E. coli isolates only. Conjugation experiments demonstrated that there was plasmid-mediated transfer of bla(CMY-2) and bla(TEM-1). These results highlighted the role of retail chicken meat as a potential source for multidrug-resistant strains of E. coli and Salmonella. To the best of our knowledge, this is the 1st report of isolation and molecular characterization of multidrug-resistant strains of E. coli from retail chicken meat in Japan.

  20. Vanillin production by recombinant strains of Escherichia coli Produção de vanilina por linhagens recombinantes de Escherichia coli

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    Attilio Converti

    2003-11-01

    Full Text Available Vanillin production from ferulate was studied using different recombinant strains of Escherichia coli. To prevent the occurrence of aerobic conditions and then possible product oxidation, tests were performed in Erlenmeyer flasks under mild mixing (150 rpm. Among other transformants, E. coli JM109(pBB1 appeared to be the best vanillin producer, being able to convert no less than 95% of starting ferulate to the product within 1h. This yield decreased down to 72% after 72h, likely because of a non-specific oxidase activity responsible for vanillin oxidation to vanillate.A produção de vanilina a partir de ácido ferúlico foi estudada utilizando-se diferentes linhagens recombinantes de Escherichia coli. Para prevenir a ocorrência de condições de aerobiose e a possível oxidação do produto, os ensaios foram realizados em frascos Erlenmeyer sob agitação moderada (150 rpm. E. coli JM109 (pBBI mostrou-se o melhor produtor de vanilina entre os demais agentes transformantes, sendo capaz de converter 95% do ácido ferúlico inicial em produto após 1h, rendimento este que decresceu para 72% após 72h, provavelmente devido à atividade de uma oxidase não-específica responsável pela oxidação de vanilina a ácido vanílico.

  1. Virulence factors and antimicrobial resistance in Escherichia coli strains isolated from hen egg shells.

    Science.gov (United States)

    Grande Burgos, María José; Fernández Márquez, Maria Luisa; Pérez Pulido, Rubén; Gálvez, Antonio; Lucas López, Rosario

    2016-12-05

    Eggs may contain extraintestinal pathogenic (ExPEC) and diarrheogenic (DEC) Escherichia coli which in addition may carry antibiotic resistance. The wide use of biocides and disinfectants in the food industry may induce biocide tolerance in bacteria. The aim of the present study was to evaluate biocide tolerance and antibiotic resistance in E. coli from hen egg shells. A total of 27 isolates obtained from a screening of 180 eggs were studied. Seven isolates carried both eae and bfpA genes of typical enteropathogenic E. coli (EPEC) strains, while 14 isolates only carried eae associated with atypical EPEC strains. Shiga toxin genes stx and stx2 were detected in four isolates. Heat-stable and heat-labile enterotoxin genes as well as aggR were also detected. Several isolates had minimum inhibitory concentrations (MICs) that were higher than the wild-type for the biocide hexadecylpyridinium chloride (HDP, 18.52%) or the commercial disinfectant P3 oxonia (OX, 14.81%). Antibiotic resistance was detected for ampicillin (37.03%), streptomycin (37.03%), tetracycline (37.03%), chloramphenicol (11.11%), nalidixic acid (18.51%) and trimethoprim-sulfamethoxazole (14.81%). Eight isolates (29.63%) were biocide tolerant and antibiotic resistant. Efflux pump genes detected included acrB (96.29%), mdfA (85.18%) and oxqA (37.03%), in addition to quaternary ammonium compound (QAC) resistance genes qacA/B (11.11%) and qacE (7.40%). Antibiotic resistance genes detected included blaCTX-M-2 (22.22%), blaTEM (3.70%), blaPSE (3.70%), tet(A) (29.63%), tet(B) (29.63%), tet(C) (7.40%), tet(E) (11.11%), aac(6')-Ib (3.70%), sul1 (14.81%), dfrA12 (3.70%) and dfrA15 (3.70%). Most isolates (96.30%) carried more than one genetic determinant of resistance. The most frequent combinations were efflux pump components acrB and mdfA with tetracycline resistance genes (33.33% of isolates). Isolates carrying QAC resistance genes also carried between 4 and 8 of the additional antimicrobial resistance genes

  2. Photoreactivation and dark repair of environmental E. coli strains following 24 kHz continuous ultrasound and UV-C irradiation.

    Science.gov (United States)

    Kaur, Jasjeet; Karthikeyan, Raghupathy; Pillai, Suresh D

    2016-07-02

    In this study, effects of 24 kHz continuous ultrasound and UV-C on inactivation and potential repair of environmental E. coli strains were studied through a culture based method and a metabolic activity assay. Three environmental E. coli strains isolated from fecal samples of feral hog and deer and treated wastewater effluent were studied and compared with a laboratory E. coli strain (ATCC® 10798). Metabolic activity of E. coli cells during the inactivation and repair period was assessed using the AlamarBlue® assay. Transmission electron microscopy assays were also performed to evaluate morphological damage of bacterial cell wall. After 24 h of photoreactivation period, laboratory E. coli strain (ATCC® 10798) reactivated by 30% and 42% in contrast to E. coli isolate from treated wastewater effluent, which reactivated by 53% and 82% after ultrasound and UV-C treatment, respectively. Possible shearing and reduction in cell size of E. coli strains exposed to ultrasound was revealed by transmission electron micrographs. Metabolic activity of E. coli strains was greatly reduced due to morphological damage to cell membrane caused by 24 kHz continuous ultrasound. Based upon experimental data and TEM micrographs, it could be concluded that ultrasound irradiation has potential in advanced water treatment and water reuse applications.

  3. D-Alanine-Controlled Transient Intestinal Mono-Colonization with Non-Laboratory-Adapted Commensal E. coli Strain HS.

    Science.gov (United States)

    Cuenca, Miguelangel; Pfister, Simona P; Buschor, Stefanie; Bayramova, Firuza; Hernandez, Sara B; Cava, Felipe; Kuru, Erkin; Van Nieuwenhze, Michael S; Brun, Yves V; Coelho, Fernanda M; Hapfelmeier, Siegfried

    2016-01-01

    Soon after birth the mammalian gut microbiota forms a permanent and collectively highly resilient consortium. There is currently no robust method for re-deriving an already microbially colonized individual again-germ-free. We previously developed the in vivo growth-incompetent E. coli K-12 strain HA107 that is auxotrophic for the peptidoglycan components D-alanine (D-Ala) and meso-diaminopimelic acid (Dap) and can be used to transiently associate germ-free animals with live bacteria, without permanent loss of germ-free status. Here we describe the translation of this experimental model from the laboratory-adapted E. coli K-12 prototype to the better gut-adapted commensal strain E. coli HS. In this genetic background it was necessary to complete the D-Ala auxotrophy phenotype by additional knockout of the hypothetical third alanine racemase metC. Cells of the resulting fully auxotrophic strain assembled a peptidoglycan cell wall of normal composition, as long as provided with D-Ala and Dap in the medium, but could not proliferate a single time after D-Ala/Dap removal. Yet, unsupplemented bacteria remained active and were able to complete their cell cycle with fully sustained motility until immediately before autolytic death. Also in vivo, the transiently colonizing bacteria retained their ability to stimulate a live-bacteria-specific intestinal Immunoglobulin (Ig)A response. Full D-Ala auxotrophy enabled rapid recovery to again-germ-free status. E. coli HS has emerged from human studies and genomic analyses as a paradigm of benign intestinal commensal E. coli strains. Its reversibly colonizing derivative may provide a versatile research tool for mucosal bacterial conditioning or compound delivery without permanent colonization.

  4. D-Alanine-Controlled Transient Intestinal Mono-Colonization with Non-Laboratory-Adapted Commensal E. coli Strain HS.

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    Miguelangel Cuenca

    Full Text Available Soon after birth the mammalian gut microbiota forms a permanent and collectively highly resilient consortium. There is currently no robust method for re-deriving an already microbially colonized individual again-germ-free. We previously developed the in vivo growth-incompetent E. coli K-12 strain HA107 that is auxotrophic for the peptidoglycan components D-alanine (D-Ala and meso-diaminopimelic acid (Dap and can be used to transiently associate germ-free animals with live bacteria, without permanent loss of germ-free status. Here we describe the translation of this experimental model from the laboratory-adapted E. coli K-12 prototype to the better gut-adapted commensal strain E. coli HS. In this genetic background it was necessary to complete the D-Ala auxotrophy phenotype by additional knockout of the hypothetical third alanine racemase metC. Cells of the resulting fully auxotrophic strain assembled a peptidoglycan cell wall of normal composition, as long as provided with D-Ala and Dap in the medium, but could not proliferate a single time after D-Ala/Dap removal. Yet, unsupplemented bacteria remained active and were able to complete their cell cycle with fully sustained motility until immediately before autolytic death. Also in vivo, the transiently colonizing bacteria retained their ability to stimulate a live-bacteria-specific intestinal Immunoglobulin (IgA response. Full D-Ala auxotrophy enabled rapid recovery to again-germ-free status. E. coli HS has emerged from human studies and genomic analyses as a paradigm of benign intestinal commensal E. coli strains. Its reversibly colonizing derivative may provide a versatile research tool for mucosal bacterial conditioning or compound delivery without permanent colonization.

  5. Surveillance of Diarrheagenic Escherichia coli strains isolated from diarrhea cases from children, adults and elderly at northwest of Mexico

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    Adrian Canizalez-Roman

    2016-11-01

    Full Text Available Diarrheagenic Escherichia coli (DEC strains are a main cause of gastrointestinal disease in developing countries. In this study we report the epidemiologic surveillance in a four-year period (January 2011 to December 2014 of DEC strains causing acute diarrhea throughout the Sinaloa State, Mexico. DEC strains were isolated from outpatients of all ages with acute diarrhea (N=1,037. Specific DEC pathotypes were identified by PCR-amplification of genes encoding virulence factors. The adhesion phenotype and antibiotic resistance were also investigated. DEC strains were detected in 23.3% (242/1037 of cases. The most frequently DEC strain isolated was EAEC (12.2%, 126/242 followed by EPEC (5.1%, 53/242, ETEC (4.3%, 43/242 DAEC (1.4%, 15/242, STEC (0.3%, 3/242 and EIEC (0.2%, 2/242. EHEC strains were not detected. Overall DEC strains were more prevalent in children ≤ 2 years of age with EPEC strains the most common of DEC pathotypes. While 65% of EAEC strains were classified as typical variant based on the aggregative adherence to in vitro cultures of HEp-2 cells, a high proportion of EPEC strains was classified as atypical strains. EAEC, EPEC, ETEC and DAEC strains were distributed in the north, central and south regions of Sinaloa state. Among all DEC strains, >90% were resistant to at least one commonly prescribed antibiotic. Strains were commonly resistant to first-line antibiotics such as tetracycline, ampicillin and sulfamethoxazole-trimethoprim. Furthermore, more than 80% of DEC isolates were multi-drug resistant and EPEC and DAEC were the categories with major proportion of this feature. In conclusion, in nearly one out of four cases of acute diarrhea in Northwestern Mexico a multi-drug resistant DEC strain was isolated, in these cases EAEC was the most prevalent (52% pathotype.

  6. [Characterization of first sorbitol-fermenting shiga toxin-producing Escherichia coli O157:H- strain isolated in Poland].

    Science.gov (United States)

    Jakubczak, Aleksandra; Szych, Jolanta; Januszkiewicz, Kamil

    2008-01-01

    Sorbitol-fermenting shiga toxin-producing E. coli O157:H- strains have emerged as a cause of human disease in many European and non-European countries. The role of SF VTEC O157:H- in the etiology of pediatric HUS and diarrhea is significant. We characterized the first SF VTEC O157:H- strain isolated from 9 year old patient in Poland. Strain possessed many traits characteristics for SF VTEC O157:H-. It fermented sorbitol after overnight incubation and produced beta-glucuronidase. It possessed the stx2, eae-gamma, EhlyA and sfpA genes and did not harbour plasmid-encoded katP and espP genes. Motility was not expressed but the strain possessed the chromosomal fliC locus for H7 antigen. The spread of SF VTEC O157:H- strains demonstrates the need for appropriate procedures for their microbiological diagnosis in Poland.

  7. Detergents enhance EspB secretion from Escherichia coli strains harboring the locus for the enterocyte effacement (LEE) gene.

    Science.gov (United States)

    Nakasone, Noboru; Toma, Claudia; Higa, Naomi; Koizumi, Yukiko; Ogura, Yasunori; Suzuki, Toshihiko

    2011-02-01

    The effects of detergents (cholic acid, deoxycholic acid, Triton X-100, and Nonidet P-40) on the secretion of EspB from the locus for enterocyte effacement (LEE) gene-positive Escherichia coli strains were examined. Clinical isolates of eight EPEC strains and seven STEC strains were used to detect EspB after they had been cultivated in Luria-Bertani (LB) broth containing one of the detergents. When the bacteria were cultured in LB broth supplemented with one of the detergents, the amount of EspB produced was increased by 2-32-fold depending on the detergent and the strain used. EspB was detected in all strains when they were cultured in LB broth containing all of the detergents. The results obtained in this study can be applied to immunological diagnostic methods for detecting EspB and also to the production of EspB for research purposes.

  8. Fate and persistence of a pathogenic NDM-1-positive Escherichia coli strain in anaerobic and aerobic sludge microcosms

    KAUST Repository

    Mantilla-Calderon, David

    2017-04-15

    The presence of emerging biological pollutants in treated wastewater effluents has gained attention due to increased interest in water reuse. To evaluate the effectiveness of the removal of such contaminants by the conventional wastewater treatment process, the fate and decay kinetics of NDM-1-positive Escherichia coli strain PI7 and its plasmid-encoded antibiotic resistance genes (ARGs) were assessed in microcosms of anaerobic and aerobic sludge. Results showed that E. coli PI7 decayed at a significantly slower rate under anaerobic conditions. Approximate half-lives were 32.4 ± 1.4 h and 5.9 ± 0.9 h in the anaerobic and aerobic microcosms, respectively. In the aerobic microcosms, after 72 h of operation, E. coli PI7 remained detectable but no further decay was observed. Instead, 1 in every 10000 E. coli cells was identified to be recalcitrant to decay and persist indefinitely in the sludge. ARGs associated with the E. coli PI7 were detected to have transferred to other native microorganisms in the sludge, or are released to the liquid fraction upon host decay. Extracellular DNA quickly degraded in the liquid fraction of the aerobic sludge. In contrast, no DNA decay was detected in the anaerobic sludge water matrix throughout the 24 h sampling period. This study suggests an increased likelihood of environmental dispersion of ARGs associated with anaerobically treated wastewater effluents and highlights the potential importance of persister cells in the dissemination of E. coli in the environment during reuse events of treated wastewater.IMPORTANCE This study examines the decay kinetics of a pathogenic and antibiotic resistant strain of Escherichia coli in microcosms simulating biological treatment units of aerobic and anaerobic sludge. The results of this study points at a significantly prolonged persistence of the E. coli and the associated antibiotic resistance gene in the anaerobic sludge. However, horizontal transfer of the plasmid encoding the antibiotic

  9. Dissemination of cephalosporin resistance genes between Escherichia coli strains from farm animals and humans by specific plasmid lineages.

    Directory of Open Access Journals (Sweden)

    Mark de Been

    2014-12-01

    Full Text Available Third-generation cephalosporins are a class of β-lactam antibiotics that are often used for the treatment of human infections caused by Gram-negative bacteria, especially Escherichia coli. Worryingly, the incidence of human infections caused by third-generation cephalosporin-resistant E. coli is increasing worldwide. Recent studies have suggested that these E. coli strains, and their antibiotic resistance genes, can spread from food-producing animals, via the food-chain, to humans. However, these studies used traditional typing methods, which may not have provided sufficient resolution to reliably assess the relatedness of these strains. We therefore used whole-genome sequencing (WGS to study the relatedness of cephalosporin-resistant E. coli from humans, chicken meat, poultry and pigs. One strain collection included pairs of human and poultry-associated strains that had previously been considered to be identical based on Multi-Locus Sequence Typing, plasmid typing and antibiotic resistance gene sequencing. The second collection included isolates from farmers and their pigs. WGS analysis revealed considerable heterogeneity between human and poultry-associated isolates. The most closely related pairs of strains from both sources carried 1263 Single-Nucleotide Polymorphisms (SNPs per Mbp core genome. In contrast, epidemiologically linked strains from humans and pigs differed by only 1.8 SNPs per Mbp core genome. WGS-based plasmid reconstructions revealed three distinct plasmid lineages (IncI1- and IncK-type that carried cephalosporin resistance genes of the Extended-Spectrum Beta-Lactamase (ESBL- and AmpC-types. The plasmid backbones within each lineage were virtually identical and were shared by genetically unrelated human and animal isolates. Plasmid reconstructions from short-read sequencing data were validated by long-read DNA sequencing for two strains. Our findings failed to demonstrate evidence for recent clonal transmission of

  10. Frequency of Extended-Spectrum Beta-lactamases (ESBLs) in strains of Klebsiella and E. coli isolated from patients hospitalized in Yazd.

    Science.gov (United States)

    Zandi, Hengameh; Tabatabaei, Seyed Mostafa; Ehsani, Fatemeh; Zarch, Mojtaba Babaei; Doosthosseini, Samira

    2017-02-01

    Frequency of extended-spectrum beta-lactamases (ESBLs) and its variants may vary in different geographical areas, as reports indicate their spread in some certain communities. The aim of this study was to determine the frequency of ESBLs in strains of Klebsiella and E. coli, isolated from patients hospitalized in teaching hospitals of Yazd. This cross-sectional study was carried out on samples including E. coli and Klebsiella strains collected from laboratories of Shahid Sadoughi and Shahid Rahnemoun hospitals in Yazd, Iran in the period of 2011-2012. The colonies which were positive in lactose Eosin methylene-blue (EMB) medium were identified by biochemical methods, and 270 strains of Klebsiella and E. coli were isolated. Collected data and information were analyzed using Fisher's exact test and descriptive statistics such as mean in SPSS software, version 15, at a significant level of 0.05. In this study, 270 samples were examined, including 152 samples of E. coli (56.3%) and 118 samples of Klebsiella pneumonia (43.7%). Among the 152 samples of E. coli, 45 strains (30%) were producers of ESBLs. In addition, among the 118 samples of Klebsiella pneumonia, 44 strains (37.3%) were producers of ESBLs. E. coli strains showed the most resistance to Cefotaxime (100%), Ceftazidime (97.7%), and Cefepime (75.5%) respectively and Klebsiella strains showed the most resistance to Cefotaxime (100%), Ceftazidime (100%) and Cefepime (79.5%), respectively. Frequency of ESBLs in Klebsiella strains was higher than E. coli strains. No significant relationship was found between frequency of ESBLs and age or gender. In addition, E. coli strains showed the highest sensitivity to Imipenem, Amoxicillin/clavulanate, and Ciprofloxacin, while the highest antibiotic sensitivity of Klebsiella strains was shown to be to Piperacillin, Imipenem, and Amoxicillin/clavulanate.

  11. Effects of the Probiotic Enterococcus faecium and Pathogenic Escherichia coli Strains in a Pig and Human Epithelial Intestinal Cell Model

    Directory of Open Access Journals (Sweden)

    Ulrike Lodemann

    2015-01-01

    Full Text Available The aim of this study has been to elucidate the effect of the probiotic Enterococcus faecium NCIMB 10415 on epithelial integrity in intestinal epithelial cells and whether pre- and coincubation with this strain can reproducibly prevent damage induced by enterotoxigenic (ETEC and enteropathogenic Escherichia coli (EPEC. Porcine (IPEC-J2 and human (Caco-2 intestinal epithelial cells were incubated with bacterial strains and epithelial integrity was assessed by measuring transepithelial electrical resistance (TEER and mannitol flux rates. E. faecium alone increased TEER of Caco-2 cells without affecting mannitol fluxes whereas the E. coli strains decreased TEER and concomitantly increased mannitol flux rates in both cell lines. Preincubation with E. faecium had no effect on the TEER decrease induced by E. coli in preliminary experiments. However, in a second set of experiments using a slightly different protocol, E. faecium ameliorated the TEER decrease induced by ETEC at 4 h in IPEC-J2 and at 2, 4, and 6 h in Caco-2 cells. We conclude that E. faecium positively affected epithelial integrity in monoinfected Caco-2 cells and could ameliorate the damage on TEER induced by an ETEC strain. Reproducibility of the results is, however, limited when experiments are performed with living bacteria over longer periods.

  12. Succinate overproduction: A case study of computational strain design using a comprehensive Escherichia coli kinetic model

    Directory of Open Access Journals (Sweden)

    Ali eKhodayari

    2015-01-01

    Full Text Available Computational strain design prediction accuracy has been the focus for many recent efforts through the selective integration of kinetic information into metabolic models. In general, kinetic model prediction quality is determined by the range and scope of genetic and/or environmental perturbations used during parameterization. In this effort, we apply the k-OptForce procedure on a kinetic model of E. coli core metabolism constructed using the Ensemble Modeling (EM method and parameterized using multiple mutant strains data under aerobic respiration with glucose as the carbon source. Minimal interventions are identified that improve succinate yield under both aerobic and anaerobic conditions to test the fidelity of model predictions under both genetic and environmental perturbations. Under aerobic condition, k-OptForce identifies interventions that match existing experimental strategies pointing at a number of unexplored flux redirections such as routing glyoxylate flux through the glycerate metabolism to improve succinate yield. Many of the identified interventions rely on the kinetic descriptions and would not be discoverable by a purely stoichiometric description. In contrast, under fermentative (anaerobic conditions, k-OptForce fails to identify key interventions including up-regulation of anaplerotic reactions and elimination of competitive fermentative products. This is due to the fact that the pathways activated under anaerobic conditions were not properly parameterized as only aerobic flux data were used in the model construction. This study shed light on the importance of condition-specific model parameterization and provides insight onto how to augment kinetic models so as to correctly respond to multiple environmental perturbations.

  13. Succinate Overproduction: A Case Study of Computational Strain Design Using a Comprehensive Escherichia coli Kinetic Model.

    Science.gov (United States)

    Khodayari, Ali; Chowdhury, Anupam; Maranas, Costas D

    2014-01-01

    Computational strain-design prediction accuracy has been the focus for many recent efforts through the selective integration of kinetic information into metabolic models. In general, kinetic model prediction quality is determined by the range and scope of genetic and/or environmental perturbations used during parameterization. In this effort, we apply the k-OptForce procedure on a kinetic model of E. coli core metabolism constructed using the Ensemble Modeling (EM) method and parameterized using multiple mutant strains data under aerobic respiration with glucose as the carbon source. Minimal interventions are identified that improve succinate yield under both aerobic and anaerobic conditions to test the fidelity of model predictions under both genetic and environmental perturbations. Under aerobic condition, k-OptForce identifies interventions that match existing experimental strategies while pointing at a number of unexplored flux re-directions such as routing glyoxylate flux through the glycerate metabolism to improve succinate yield. Many of the identified interventions rely on the kinetic descriptions that would not be discoverable by a purely stoichiometric description. In contrast, under fermentative (anaerobic) condition, k-OptForce fails to identify key interventions including up-regulation of anaplerotic reactions and elimination of competitive fermentative products. This is due to the fact that the pathways activated under anaerobic condition were not properly parameterized as only aerobic flux data were used in the model construction. This study shed light on the importance of condition-specific model parameterization and provides insight on how to augment kinetic models so as to correctly respond to multiple environmental perturbations.

  14. 60株大肠杆菌的分-离与致病性鉴定%Isolation and Identification of 60 Pathogenic Escherichia coli Strains

    Institute of Scientific and Technical Information of China (English)

    黄宗梅; 陈红英; 崔沛; 韩玉林; 李新生

    2011-01-01

    大肠杆菌(Escherichia coli,E coli)是鸡肠道常在菌,部分菌株具有致病性.鸡大肠杆菌病就是由大肠杆菌中的某些致病性菌株引起的鸡感染性疾病.致病性大肠杆菌均携带毒力岛基因ChuA.为鉴定分离自临床病例的鸡大肠杆菌的致病性,本试验采用PCR法检测ChuA基因进行分子生物学鉴定,60株大肠杆菌中有31株属于致病性大肠杆菌,总阳性率为51.67%.%Escherichia coli (E. coli) is often in chieken intestinal. Only some strains are pathogenic for chicken. Chicken Escherichia coli disease is caused by these pathogenic E. coli strains,and they have pathogenicity island genes ChuA. In order to identify 60 E. coli strains isolated from diseased chicken,ChuA gene was tested by PCR. 31 E. coli strains are pathogenic.The total positive rate is 5l. 67%.

  15. Antimicrobial Susceptibility of Escherichia coli Strains Isolated from Alouatta spp. Feces to Essential Oils

    Directory of Open Access Journals (Sweden)

    Valéria Maria Lara

    2016-01-01

    Full Text Available This study evaluated the in vitro antibacterial activity of essential oils from Lippia graveolens (Mexican oregano, Origanum vulgaris (oregano, Thymus vulgaris (thyme, Rosmarinus officinalis (rosemary, Cymbopogon nardus (citronella, Cymbopogon citratus (lemongrass, and Eucalyptus citriodora (eucalyptus against Escherichia coli (n=22 strains isolated from Alouatta spp. feces. Minimum inhibitory concentration (MIC and minimum bactericidal concentration (MBC were determined for each isolate using the broth microdilution technique. Essential oils of Mexican oregano (MIC mean = 1818 μg mL−1; MBC mean = 2618 μg mL−1, thyme (MIC mean = 2618 μg mL−1; MBC mean = 2909 μg mL−1, and oregano (MIC mean = 3418 μg mL−1; MBC mean = 4800 μg mL−1 showed the best antibacterial activity, while essential oils of eucalyptus, rosemary, citronella, and lemongrass displayed no antibacterial activity at concentrations greater than or equal to 6400 μg mL−1. Our results confirm the antimicrobial potential of some essential oils, which deserve further research.

  16. Antimicrobial Susceptibility of Escherichia coli Strains Isolated from Alouatta spp. Feces to Essential Oils.

    Science.gov (United States)

    Lara, Valéria Maria; Carregaro, Adriano Bonfim; Santurio, Deise Flores; de Sá, Mariangela Facco; Santurio, Janio Moraes; Alves, Sydney Hartz

    2016-01-01

    This study evaluated the in vitro antibacterial activity of essential oils from Lippia graveolens (Mexican oregano), Origanum vulgaris (oregano), Thymus vulgaris (thyme), Rosmarinus officinalis (rosemary), Cymbopogon nardus (citronella), Cymbopogon citratus (lemongrass), and Eucalyptus citriodora (eucalyptus) against Escherichia coli (n = 22) strains isolated from Alouatta spp. feces. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined for each isolate using the broth microdilution technique. Essential oils of Mexican oregano (MIC mean = 1818 μg mL(-1); MBC mean = 2618 μg mL(-1)), thyme (MIC mean = 2618 μg mL(-1); MBC mean = 2909 μg mL(-1)), and oregano (MIC mean = 3418 μg mL(-1); MBC mean = 4800 μg mL(-1)) showed the best antibacterial activity, while essential oils of eucalyptus, rosemary, citronella, and lemongrass displayed no antibacterial activity at concentrations greater than or equal to 6400 μg mL(-1). Our results confirm the antimicrobial potential of some essential oils, which deserve further research.

  17. Antibiotic Resistance in Escherichia Coli Strains Isolated from Urine of Inpatients and Outpatients

    Directory of Open Access Journals (Sweden)

    Abolfazl Davoodabadi

    2012-08-01

    Full Text Available The urinary tract infections regarded as a health problem around the world and not only as an agent of nosocomial infections but also infections in the community. Community acquired UTIs cause significant illness in the first 2 years of life [1]. Urinary tract infections in both inpatient and outpatient are common and widespread use of antibiotics is often the cause of emerging one or more antibiotic-resistant microorganisms [2]. Most studies have shown higher antibiotic resistance in bacterial strains isolated from hospitalized patients than outpatients. In this study, antibiogram was performed using disk diffusion susceptibility method according to NCCLS standards of the International Committee [3]. 8 different antibiotics, including ciprofloxacin (CP: 30 μg, ceftriaxone (CRO: 30 μg, cephalotin (CF: 30 μg, cefixime (CFM: 5 μg, cotrimoxazole (SXT, nalidixic acid (NA: 30 μg, nitrofurantoin (FM: 300 μg, gentamicin (GM: 10 μg were used for antibiogram. During 1388 the total number of urine samples sent to hospital microbiology laboratories valiasr (aj of Arak was 5156, of which 446 samples (65.8% were positive for E. coli culture.

  18. Escherichia coli W as a new platform strain for the enhanced production of L-valine by systems metabolic engineering.

    Science.gov (United States)

    Park, Jin Hwan; Jang, Yu-Sin; Lee, Jeong Wook; Lee, Sang Yup

    2011-05-01

    A less frequently employed Escherichia coli strain W, yet possessing useful metabolic characteristics such as less acetic acid production and high L-valine tolerance, was metabolically engineered for the production of L-valine. The ilvA gene was deleted to make more pyruvate, a key precursor for L-valine, available for enhanced L-valine biosynthesis. The lacI gene was deleted to allow constitutive expression of genes under the tac or trc promoter. The ilvBN(mut) genes encoding feedback-resistant acetohydroxy acid synthase (AHAS) I and the L-valine biosynthetic ilvCED genes encoding acetohydroxy acid isomeroreductase, dihydroxy acid dehydratase, and branched chain amino acid aminotransferase, respectively, were amplified by plasmid-based overexpression. The global regulator Lrp and L-valine exporter YgaZH were also amplified by plasmid-based overexpression. The engineered E. coli W (ΔlacI ΔilvA) strain overexpressing the ilvBN(mut) , ilvCED, ygaZH, and lrp genes was able to produce an impressively high concentration of 60.7 g/L L-valine by fed-batch culture in 29.5 h, resulting in a high volumetric productivity of 2.06 g/L/h. The most notable finding is that there was no other byproduct produced during L-valine production. The results obtained in this study suggest that E. coli W can be a good alternative to Corynebacterium glutamicum and E. coli K-12, which have so far been the most efficient L-valine producer. Furthermore, it is expected that various bioproducts including other amino acids might be more efficiently produced by this revisited platform strain of E. coli.

  19. Fatores de virulência em linhagens de Escherichia coli isoladas de mastite bovina Virulence factors in Escherichia coli strains isolated from bovine mastitis

    Directory of Open Access Journals (Sweden)

    M.G. Ribeiro

    2006-10-01

    Full Text Available Avaliou-se a ocorrência de fatores de virulência e do sorotipo O157:H7 em 120 linhagens de Escherichia coli, isoladas de 80 casos de mastite clínica bovina e 40 de mastite subclínica. Verificou-se alfa-hemolisina em oito (6,7% linhagens, isoladas de cinco casos de mastite clínica e três de mastite subclínica e em nenhuma das estirpes detectou-se enteroemolisina. A presença de sideróforos foi encontrada em 11 (9,2% linhagens, sete de mastite clínica e quatro de subclínica. Em duas (1,7% estirpes isoladas de mastite subclínica, identificou-se enterotoxina STa. Observou-se efeito citopático em células vero compatível com a produção de verotoxina-VT em cinco (4,2% linhagens, duas de mastite clínica e três subclínicas. Em uma (0,8% linhagem isolada de mastite clínica, detectou-se efeito citopático compatível com o fator necrosante citotóxico. Nenhuma estirpe apresentou-se sorbitol-negativa no MacConkey-sorbitol, tampouco aglutinou com o sorotipo O157:H7. Os antimicrobianos mais efetivos foram polimixina B (97,5% e norfloxacina (95,8%. Observou-se multi-resistência a dois ou mais antimicrobianos em 24 (20% estirpes, principalmente com o uso de ampicilina e ceftiofur.The occurrence of different virulence factors and O157:H7 serotype investigation in 120 Escherichia coli strains isolated from clinical (80 cases and subclinical (40 cases bovine mastitis was evaluated. Alpha-haemolysin was detected in 8 (6.7% strains (5 clinical and 3 subclinical cases. None strain showed enterohaemolysin production. E. coli growth under iron restriction conditions (siderophores production was observed in 11 (9.2% strains (7 clinical and 4 subclinical cases. STa enterotoxin was detected in 2 (1.7% strains from subclinical cases. Cytotoxic effect in vero cells compatible with verotoxin-VT production was observed in 5 (4.2% strains (2 clinical and 3 subclinical cases. One strain (0.8% isolated from clinical mastitis showed cytophatic effect in vero

  20. Asymptomatic bacteriuria Escherichia coli strain 83972 carries mutations in the foc locus and is unable to express F1C fimbriae

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Schembri, M.A.; Ulett, G.C.

    2006-01-01

    this adhesin. The data imply that E. coli 83972 has lost its ability to express this important colonization factor as a result of host-driven evolution. The ancestor of the strain seems to have been a pyelonephritis strain of phylogenetic group B2. Strain 83972 therefore represents an example of bacterial...

  1. Asymptomatic bacteriuria Escherichia coli strain 83972 carries mutations in the foc locus and is unable to express F1C fimbriae

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Schembri, M.A.; Ulett, G.C.

    2006-01-01

    this adhesin. The data imply that E. coli 83972 has lost its ability to express this important colonization factor as a result of host-driven evolution. The ancestor of the strain seems to have been a pyelonephritis strain of phylogenetic group B2. Strain 83972 therefore represents an example of bacterial...

  2. The prevalence of O serogroups of Escherichia coli strains causing acute urinary tract infection in children in Iran

    Directory of Open Access Journals (Sweden)

    Fatemeh Emamghorashi

    2011-01-01

    Full Text Available The aim of present study was to determine the prevalence of O serogroups of Escherichia coli (E. coli strains that cause community-acquired urinary tract infections (UTI in children. In this study, 96 children with UTI referred to two Jahrom University-affiliated Hospitals in Iran were enrolled, during the period from August 2005 to August 2006. Drug sensitivity was tested by disk diffusion method and serotyping done by slide agglutination method. A total of 96 E. coli strains were isolated from urine samples of the study children whose age ranged from one month to 14 years. Cystitis was diagnosed in 49.2% and pyelonephritis in 50.8% of the study patients. Maximum drug resistance was seen with ampicilin (80.2% and the least with imipenem (1.1%. The most common type of O antigen was O1 (12.2%. There was significant correlation between the presence of O antigens and sensitivity to nalidixic acid and gentamicin (P < 0.05. This is the first report of E. coli serotyping in children with UTI from the south of Iran and their relation to antibiotic resistance and clinical presentation. Further studies from other parts of Iran and on other serotypes are recommended.

  3. Extended-spectrum beta-lactamase-producing Escherichia coli infections in children: are community-acquired strains different from nosocomial strains?

    Science.gov (United States)

    Morgand, Marjolaine; Vimont, Sophie; Bleibtreu, Alexandre; Boyd, Anders; Thien, Hoang Vu; Zahar, Jean-Ralph; Denamur, Erick; Arlet, Guillaume

    2014-11-01

    Infections caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli are an important cause of morbidity and mortality, especially in children. We compared 58 epidemiologically unrelated ESBL-producing E. coli strains that caused infections. They were isolated between 2008 and 2012 in two Parisian pediatric hospitals and grouped according to their origin into either community-acquired (CA) (n=37) or nosocomially acquired (NA) (n=21) strains. Molecular characteristics of the ESBLs, phylogenetic traits of the strains including their belonging to clone O25b-ST131, prevalence of associated virulence genes, growth capacities in different media, metabolic phenotype and biofilm formation abilities were studied. ESBL type, associated resistance and distribution of phylogenetic groups were similar in the CA and NA groups. More than 60% of the B2 phylogroup strains in both groups belonged to the ST131 clone. Interestingly, CA strains possessed more genes encoding virulence factors and the distribution of these genes differed significantly between the two groups: fyuA, hlyC, papC and papGII were more frequent in the CA group, whereas iroN was more frequent in the NA group. CA strains also showed enhanced growth capacities in Luria Bertani rich medium. They tended to produce more biofilm but the difference was not significant. This study confirms the wide spread of clone ST131 among infected children, regardless of whether their infections were community- or nosocomially acquired. It highlights genotypic and phenotypic differences according to the origin of the strains that could indicate adaptability of these multi-resistant bacteria to specific environmental and host factors. Copyright © 2014 Elsevier GmbH. All rights reserved.

  4. Restoration of growth by manganese in a mutant strain of Escherichia coli lacking most known iron and manganese uptake systems

    DEFF Research Database (Denmark)

    Taudte, Nadine; German, Nadezhda; Zhu, Yong-Guan

    2016-01-01

    The interplay of manganese and iron homeostasis and oxidative stress in Escherichia coli can give important insights into survival of bacteria in the phagosome and under differing iron or manganese bioavailabilities. Here, we characterized a mutant strain devoid of all know iron/manganese......-uptake systems relevant for growth in defined medium. Based on these results an exit strategy enabling the cell to cope with iron depletion and use of manganese as an alternative for iron could be shown. Such a strategy would also explain why E. coli harbors some iron- or manganese-dependent iso......-enzymes such as superoxide dismutases or ribonucleotide reductases. The benefits for gaining a means for survival would be bought with the cost of less efficient metabolism as indicated in our experiments by lower cell densities with manganese than with iron. In addition, this strain was extremely sensitive to the metalloid...

  5. Complete genome sequence of DSM 30083(T), the type strain (U5/41(T)) of Escherichia coli, and a proposal for delineating subspecies in microbial taxonomy.

    Science.gov (United States)

    Meier-Kolthoff, Jan P; Hahnke, Richard L; Petersen, Jörn; Scheuner, Carmen; Michael, Victoria; Fiebig, Anne; Rohde, Christine; Rohde, Manfred; Fartmann, Berthold; Goodwin, Lynne A; Chertkov, Olga; Reddy, Tbk; Pati, Amrita; Ivanova, Natalia N; Markowitz, Victor; Kyrpides, Nikos C; Woyke, Tanja; Göker, Markus; Klenk, Hans-Peter

    2014-01-01

    Although Escherichia coli is the most widely studied bacterial model organism and often considered to be the model bacterium per se, its type strain was until now forgotten from microbial genomics. As a part of the G enomic E ncyclopedia of B acteria and A rchaea project, we here describe the features of E. coli DSM 30083(T) together with its genome sequence and annotation as well as novel aspects of its phenotype. The 5,038,133 bp containing genome sequence includes 4,762 protein-coding genes and 175 RNA genes as well as a single plasmid. Affiliation of a set of 250 genome-sequenced E. coli strains, Shigella and outgroup strains to the type strain of E. coli was investigated using digital DNA:DNA-hybridization (dDDH) similarities and differences in genomic G+C content. As in the majority of previous studies, results show Shigella spp. embedded within E. coli and in most cases forming a single subgroup of it. Phylogenomic trees also recover the proposed E. coli phylotypes as monophyla with minor exceptions and place DSM 30083(T) in phylotype B2 with E. coli S88 as its closest neighbor. The widely used lab strain K-12 is not only genomically but also physiologically strongly different from the type strain. The phylotypes do not express a uniform level of character divergence as measured using dDDH, however, thus an alternative arrangement is proposed and discussed in the context of bacterial subspecies. Analyses of the genome sequences of a large number of E. coli strains and of strains from > 100 other bacterial genera indicate a value of 79-80% dDDH as the most promising threshold for delineating subspecies, which in turn suggests the presence of five subspecies within E. coli.

  6. Effect of solar radiation on multidrug resistant E. coli strains and antibiotic mixture photodegradation in wastewater polluted stream.

    Science.gov (United States)

    Rizzo, L; Fiorentino, A; Anselmo, A

    2012-06-15

    The effect of solar radiation on the inactivation of multidrug resistant Escherichia coli (MDR) strains selected from an urban wastewater treatment plant (UWWTP) effluent and the change of their resistance to a mixture of three antibiotics (evaluated in terms of minimum inhibit concentration (MIC)) in wastewater polluted stream were investigated. The solar photodegradation of the mixture of the three target antibiotics (amoxicillin (AMX), ciprofloxacin (CPX), and sulfamethoxazole (SMZ)) was also evaluated. Additionally, since UWWTP effluents are possible sources of antibiotics and antibiotic resistant bacteria, the disinfection by conventional chlorination process of the UWWTP effluent inoculated with MDR strains was investigated too. Solar radiation poorly affected the inactivation of the two selected antibiotic resistant E. coli strains (40 and 60% after 180 min irradiation). Moreover, solar radiation did not affect strain resistance to AMX (MIC>256 μg/mL) and SMZ (MIC>1024 μg/mL), but affected resistance of the lower resistance strain to CPX (MIC decreased by 33% but only after 180 min of irradiation). Chlorination of wastewater sample strongly decreased the number of the two selected antibiotic resistant E. coli strains (99.667 and 99.999%), after 60 min of contact time at 2.0 mg/L initial chlorine concentration, but the resistance of survived colonies to antibiotics was unchanged. Finally, the solar photodegradation rate of the antibiotic mixture (1mg/L initial concentration respectively) resulted in the following order (half-life time): CPX (t(1/2)=24 min)resistance to SMZ in surface water is significantly higher compared to CPX and AMX. Copyright © 2012 Elsevier B.V. All rights reserved.

  7. Molecular Analysis of Asymptomatic Bacteriuria Escherichia coli Strain VR50 Reveals Adaptation to the Urinary Tract by Gene Acquisition

    DEFF Research Database (Denmark)

    Beatson, Scott A.; Ben Zakour, Nouri L.; Totsika, Makrina;

    2015-01-01

    to adhere to human bladder epithelial cells. In the mouse model of UTI, VR50afa and VR50afaE displayed reduced bladder colonization compared to wild-type VR50, similar to the colonization level of the GI-VR50-pheV mutant. Our study suggests that E. coli VR50 is a commensal-like strain that has acquired...

  8. Survey on the occurrence of shiga toxin producing strains of Escherichia coli (STEC) in drinking water from central Hessia

    Energy Technology Data Exchange (ETDEWEB)

    Stelz, A.; Zschoeck, M. [Untersuchungsamt Hessen, Giessen (Germany); Fels, K.; Alter, E. [Fachhochschule Giessen-Friedberg, Giessen (Germany)

    2001-07-01

    Consumption of unheated bovine food is said to be the main cause of outbreaks of enterohemorrhagic colitis (EC) and hemolytic syndrome (HUS) by E. coli O157 in man. Some cases were attributed to the ingestion of contaminated drinking water. Certain types of resources for drinking water supply are at risk of contamination with cattle faeces or sewage. In routine drinking water control E. coli is an index organism and must not be found in a sample of 100 ml. We investigated a total of 1700 drinking water samples from central Hessia (Germany) for shiga toxin producing strains of E. coli (STEC). A developed sensitive investigation scheme made it possible to detect up to 10 STEC per 100 ml water sample at least. ELISA was used for detection of shiga toxins 1 and 2, m-PCR for the genes encoding stx1, stx2, EHEC-hlyA and eaeA, plating on Enterohemolysin agar for detection of the enterohemolysin phenotype and slide agglutination technique for confirming E. coli O157. In 108 out of 1700 drinking water samples E. coli was detected, but none of the isolates belonged to serotype O157. ST1 and ST2 were not produced by the isolates. Thus, under normal circumstances drinking water in central Hessia seems to be no vehicle for STEC. (orig.)

  9. Multidrug Resistance in Escherichia coli Strains Isolated from Infections in Dogs and Cats in Poland (2007–2013

    Directory of Open Access Journals (Sweden)

    Magdalena Rzewuska

    2015-01-01

    Full Text Available The antimicrobial susceptibility of Escherichia coli isolates associated with various types of infections in dogs and cats was determined. The studied isolates were most frequently susceptible to fluoroquinolones and the extended-spectrum cephalosporins (ESCs, antimicrobials commonly used in treatment of infections in companion animals. However, an increase in the percentage of strains resistant to β-lactam antibiotics including ESCs was noted between January 2007 and December 2013. The frequency of multidrug-resistant (MDR E. coli isolation (66.8% of isolates is alarming. Moreover, the statistically significant increase of the percentage of MDR isolates was observed during the study period. No difference in the prevalence of multidrug resistance was found between bacteria causing intestinal and extraintestinal infections and between canine and feline isolates. Nonhemolytic E. coli isolates were MDR more often than hemolytic ones. Our study showed the companion animals in Poland as an important reservoir of MDR bacteria. These results indicate that continuous monitoring of canine and feline E. coli antimicrobial susceptibility is required. Furthermore, introduction and application of recommendations for appropriate use of antimicrobials in small animal practice should be essential to minimize the emergence of multidrug resistance among E. coli in companion animals.

  10. Multidrug Resistance in Escherichia coli Strains Isolated from Infections in Dogs and Cats in Poland (2007–2013)

    Science.gov (United States)

    Rzewuska, Magdalena; Czopowicz, Michał; Kizerwetter-Świda, Magdalena; Chrobak, Dorota; Błaszczak, Borys; Binek, Marian

    2015-01-01

    The antimicrobial susceptibility of Escherichia coli isolates associated with various types of infections in dogs and cats was determined. The studied isolates were most frequently susceptible to fluoroquinolones and the extended-spectrum cephalosporins (ESCs), antimicrobials commonly used in treatment of infections in companion animals. However, an increase in the percentage of strains resistant to β-lactam antibiotics including ESCs was noted between January 2007 and December 2013. The frequency of multidrug-resistant (MDR) E. coli isolation (66.8% of isolates) is alarming. Moreover, the statistically significant increase of the percentage of MDR isolates was observed during the study period. No difference in the prevalence of multidrug resistance was found between bacteria causing intestinal and extraintestinal infections and between canine and feline isolates. Nonhemolytic E. coli isolates were MDR more often than hemolytic ones. Our study showed the companion animals in Poland as an important reservoir of MDR bacteria. These results indicate that continuous monitoring of canine and feline E. coli antimicrobial susceptibility is required. Furthermore, introduction and application of recommendations for appropriate use of antimicrobials in small animal practice should be essential to minimize the emergence of multidrug resistance among E. coli in companion animals. PMID:25667937

  11. Multidrug resistance in Escherichia coli strains isolated from infections in dogs and cats in Poland (2007-2013).

    Science.gov (United States)

    Rzewuska, Magdalena; Czopowicz, Michał; Kizerwetter-Świda, Magdalena; Chrobak, Dorota; Błaszczak, Borys; Binek, Marian

    2015-01-01

    The antimicrobial susceptibility of Escherichia coli isolates associated with various types of infections in dogs and cats was determined. The studied isolates were most frequently susceptible to fluoroquinolones and the extended-spectrum cephalosporins (ESCs), antimicrobials commonly used in treatment of infections in companion animals. However, an increase in the percentage of strains resistant to β-lactam antibiotics including ESCs was noted between January 2007 and December 2013. The frequency of multidrug-resistant (MDR) E. coli isolation (66.8% of isolates) is alarming. Moreover, the statistically significant increase of the percentage of MDR isolates was observed during the study period. No difference in the prevalence of multidrug resistance was found between bacteria causing intestinal and extraintestinal infections and between canine and feline isolates. Nonhemolytic E. coli isolates were MDR more often than hemolytic ones. Our study showed the companion animals in Poland as an important reservoir of MDR bacteria. These results indicate that continuous monitoring of canine and feline E. coli antimicrobial susceptibility is required. Furthermore, introduction and application of recommendations for appropriate use of antimicrobials in small animal practice should be essential to minimize the emergence of multidrug resistance among E. coli in companion animals.

  12. Cefoxitin resistance mediated by loss of a porin in clinical strains of Klebsiella pneumoniae and Escherichia coli

    Directory of Open Access Journals (Sweden)

    Ananthan S

    2005-01-01

    Full Text Available PURPOSE: Porins are outer membrane protein (OMP that form water filled channels that permit the diffusion of small hydrophilic solutes like -lactam antibiotics across the outer membrane. Two major porins that facilitate diffusion of antimicrobials have been described in Klebsiella spp. and Escherichia coli. The present study was carried out to examine the role of porins among Extended Spectrum -Lactamase (ESBL and AmpC -Lactamase positive strains of Klebsiella spp. and E.coli. METHODS: Preparation of OMP from phenotypically characterized clinical isolates K.pneumoniae and E.coli and the separation of the proteins by sodium dodecyl sulfate - polyacrylamide gel electrophoresis were performed as per a previously described procedure. RESULTS: OMP analysis revealed that cefoxitin and ceftazidime resistance was mediated by loss of a porin Omp K35 in the isolates of K.pneumoniae and E.coli. CONCLUSIONS: Loss of porin mediated resistance mechanism against cefoxitin was observed among the multidrug resistant K.pneumoniae and E.coli.

  13. Resistance of E. coli strains, recovered from chickens to antibiotics with particular reference to trimethoprim-sulfamethoxazole (septrin).

    Science.gov (United States)

    Bebora, L C; Oundo, J O; Yamamoto, H

    1994-10-01

    Thirty-seven strains of E. coli recovered from cases of septicaemia in chicken were tested for sensitivity to 6 antibiotics. Minimum inhibitory concentration (MIC) determinations done on the strains showed resistance to trimethoprim-sulfamethoxazole (septrin) (100%), ampicillin (62.2%), tetracycline (51.4%), kanamycin (13.5%) and gentamicin (2.7%). All were sensitive to chloramphenicol. Conjugation studies showed easy transfer of the resistance factor for septrin to the recipient sensitive strain, K12F-, a 60 megadalton plasmid was transferred in most of the cases (a number of plasmids moved across to K12F- strains). Septrin was chosen as a referral antibiotic because it is used extensively for treating diarrhoeal cases in children in Kenya. The results expressed the possibility of the chicken being the possible source of the septrin resistance gene (plasmid) for humans, and vice versa.

  14. Diffusely adherent Escherichia coli strains expressing Afa/Dr adhesins (Afa/Dr DAEC): hitherto unrecognized pathogens.

    Science.gov (United States)

    Le Bouguénec, Chantal; Servin, Alain L

    2006-03-01

    Diffusely adherent Escherichia coli (DAEC) strains are currently considered to constitute a putative sixth group of diarrheagenic E. coli. However, on the basis of their diffuse adherence to HEp-2 and HeLa cells, the detection of afa/dra/daa-related operons encoding this adherence phenotype, and the mobilization of decay-accelerating factor, both commensal and pathogenic strains can be classified as Afa/Dr DAEC isolates. Furthermore, strains associated with diarrheal diseases and strains causing extra-intestinal infections can also be identified as Afa/Dr DAEC strains. Although several cell signaling events that occur after epithelial cells have been infected by Afa/Dr DAEC have been reported, the pathophysiological processes that allow intestinal and extra-intestinal infections to develop are not fully understood. This review focuses on the genetic organization of the afa/dra/daa-related operons and on the virulence factors that trigger cellular responses, some of which are deleterious for the host cells. Finally, this review suggests future lines of research that could help to elucidate these questions.

  15. Improvement in ethanol productivity of engineered E. coli strain SSY13 in defined medium via adaptive evolution.

    Science.gov (United States)

    Jilani, Syed Bilal; Venigalla, Siva Sai Krishna; Mattam, Anu Jose; Dev, Chandra; Yazdani, Syed Shams

    2017-07-04

    E. coli has the ability to ferment both C5 and C6 sugars and produce mixture of acids along with small amount of ethanol. In our previous study, we reported the construction of an ethanologenic E. coli strain by modulating flux through the endogenous pathways. In the current study, we made further changes in the strain to make the overall process industry friendly; the changes being (1) removal of plasmid, (2) use of low-cost defined medium, and (3) improvement in consumption rate of both C5 and C6 sugars. We first constructed a plasmid-free strain SSY13 and passaged it on AM1-xylose minimal medium plate for 150 days. Further passaging was done for 56 days in liquid AM1 medium containing either glucose or xylose on alternate days. We observed an increase in specific growth rate and carbon utilization rate with increase in passage numbers until 42 days for both glucose and xylose. The 42nd day passaged strain SSK42 fermented 113 g/L xylose in AM1 minimal medium and produced 51.1 g/L ethanol in 72 h at 89% of maximum theoretical yield with ethanol productivity of 1.4 g/L/h during 24-48 h of fermentation. The ethanol titer, yield and productivity were 49, 40 and 36% higher, respectively, for SSK42 as compared to unevolved SSY13 strain.

  16. Production of d-lactate from sugarcane bagasse and corn stover hydrolysates using metabolic engineered Escherichia coli strains.

    Science.gov (United States)

    Utrilla, José; Vargas-Tah, Alejandra; Trujillo-Martínez, Berenice; Gosset, Guillermo; Martinez, Alfredo

    2016-11-01

    In this study, the lactogenic Escherichia coli strain JU15 was used and modified to produce d-lactate (d-LA) from plant hydrolysates with a minimal nutrient addition in pH controlled fermenters. Results showed that strain JU15 produces d-LA with high yield and productivity in laboratory simulated hydrolysate media and actual sugar cane bagasse hemicellulosic hydrolysate. Strain JU15 showed sequential carbon source utilization and acetic acid production. The l-lactic and acetic acid production pathways were deleted in JU15, resulting strain AV03 (JU15 ΔpoxB, ΔackA-pta, ΔmgsA), which showed simultaneous consumption of glucose and xylose and no acetic acid production in the simulated hydrolysate. The d-LA yield from hydrolysate sugars was close to 0.95gD-LA/gsugars in all cases. Our results show that d-LA can be produced from plant hydrolysates in simple batch fermentation processes with a high productivity using engineered E. coli strains at fermenter scales from 0.2 up to 10L. Copyright © 2016 Elsevier Ltd. All rights reserved.

  17. Effects induced by XeCl laser radiation and germicidal lamp radiation on E. coli strains survival and mutability

    Science.gov (United States)

    Belloni, F.; Alifano, P.; Lorusso, A.; Monaco, C.; Nassisi, V.; Talà, A.; Tredici, M.

    2006-04-01

    In this work the mutagenic effect on Escherichia coli strains induced by UV radiation emitted by a XeCl laser (λ = 308 nm) has been analysed as a function of the exposure dose and compared with the effect induced by 254 nm radiation emitted by a conventional germicidal lamp. E. coli strains, wild-type (recA+) and mutant (recA1, defective in DNA damage repair systems), plated on LB agar, supplemented with rifampicin when requested, were irradiated by means of a germicidal lamp in the dose range 0 - 9 mJ/cm2. Similar strains were exposed to 308 nm pulsed laser radiation (τ = 20 ns FWHM; max. pulse energy: 100 mJ) in the dose range 0-1.0 x 10 4 mJ/cm2. The discrepancy between the results obtained with the lamp and the laser on the mutation frequency, suggested that the biological response to the two radiation sources involves distinct mechanisms. This hypothesis was supported by the evidence that exposure to near-UV 308 nm induced mutagenesis in the recA-defective strain at an extent considerably higher than in the recA-proficient strain.

  18. A multicopy phr-plasmid increases the ultraviolet resistance of a recA strain of Escherichia coli.

    Science.gov (United States)

    Yamamoto, K; Satake, M; Shinagawa, H

    1984-01-01

    It has been previously reported that the ultraviolet sensitivity of recA strains of Escherichia coli in the dark is suppressed by a plasmid pKY1 which carries the phr gene, suggesting that this is due to a novel effect of photoreactivating enzyme (PRE) of E. coli in the dark (Yamamoto et al., 1983a). In this work, we observed that an increase of UV-resistance by pKY1 in the dark is not apparent in strains with a mutation in either uvrA, uvrB, uvrC, lexA, recBC or recF. The sensitivity of recA lexA and recA recBC multiple mutants to UV is suppressed by the plasmid but that of recA uvrA, recA uvrB and recA uvrC is not. Host-cell reactivation of UV-irradiated lambda phage is slightly more efficient in the recA/pKY1 strain compared with the parental recA strain. On the other hand, the recA and recA/pKY1 strains do not differ significantly in the following properties: Hfr recombination, induction of lambda by UV, and mutagenesis. We suggest that dark repair of PRE is correlated with its capacity of excision repair.

  19. X-ray-induced mutations in Escherichia coli K-12 strains with altered DNA polymerase I activities

    Energy Technology Data Exchange (ETDEWEB)

    Nagata, Yuki; Kawata, Masakado; Komura, Jun-ichiro; Ono, Tetsuya; Yamamoto, Kazuo

    2003-07-25

    Spectra of ionizing radiation mutagenesis were determined by sequencing X-ray-induced endogenous tonB gene mutations in Escherichia coli polA strains. We used two polA alleles, the polA1 mutation, defective for Klenow domain, and the polA107 mutation, defective for flap domain. We demonstrated that irradiation of 75 and 50 Gy X-rays could induce 3.8- and 2.6-fold more of tonB mutation in polA1 and polA107 strains, respectively, than spontaneous level. The radiation induced spectrum of 51 tonB mutations in polA1 and 51 in polA107 indicated that minus frameshift, A:T{yields}T:A transversion and G:C{yields}T:A transversion were the types of mutations increased. Previously, we have reported essentially the same X-ray-induced tonB mutation spectra in the wild-type strain. These results indicate that (1) X-rays can induce minus frameshift, A:T{yields}T:A transversion and G:C{yields}T:A transversion in E. coli and (2) presence or absence of polymerase I (PolI) of E. coli does not have any effects on the process of X-ray mutagenesis.

  20. Antimicrobial susceptibility testing of Escherichia coli strains isolated from urinary tract infections to fluoroquinolones and detection of gyrA mutations in resistant strains

    Directory of Open Access Journals (Sweden)

    Akbari-Nakhjavani F.

    2007-05-01

    Full Text Available Widespread uses of fluoroquinolones have resulted in increasing incidences of resistance against these agents all over the world. The aim of this study was to assess, susceptibility of Escherichia coli strains from patients with Urinary Tract Infection against common fluoroquinolones and detection of mutations in the gyrA gene. Antimicrobial susceptibility testing of 164 E.coli isolates from patients with UTI, was evaluated by disk agar diffusion (DAD and MIC methods. Polymerase chain reaction of E.coli strains were performed by amplification of Quinolone Resistance Determining Region (QRDR of gyrA gene. PCR products were tested by Conformational Sensitive Gel Electrophoresis (CSGE and those with hetrodublexes were selected and examined by DNA sequencing. According to disc agar diffusion, 49.3% were resistant to nalidixic acid, 41.4% to norfloxacin, 44.5% to ofloxacin and 40.2 % to ciprofloxacin. By Minimal Inhibitory Concentration (MIC testing a high-level of resistance (42.1% to ciprofloxacin was observed. Mutations in codons 83 and 87 in all 81 isolates were positive by CSGE method.

  1. Transmission of an extended-spectrum-beta-lactamase-producing Escherichia coli (sequence type ST131) strain between a father and daughter resulting in septic shock and Emphysematous pyelonephritis.

    Science.gov (United States)

    Ender, Peter T; Gajanana, Deepakraj; Johnston, Brian; Clabots, Connie; Tamarkin, Frank J; Johnson, James R

    2009-11-01

    Probable transmission of an extended-spectrum-beta-lactamase-producing Escherichia coli strain (sequence type ST131) between a father and daughter was documented. The father developed severe, recurrent pyelonephritis with multiple small abscesses; the daughter later developed septic shock, bacteremia, and extensive emphysematous pyelonephritis. This multidrug-resistant E. coli clone appears to be highly pathogenic and transmissible.

  2. Whole-Genome Sequence of Multidrug-Resistant Campylobacter coli Strain COL B1-266, Isolated from the Colombian Poultry Chain.

    Science.gov (United States)

    Bernal, Johan F; Donado-Godoy, Pilar; Arévalo, Alejandra; Duarte, Carolina; Realpe, María E; Díaz, Paula L; Gómez, Yolanda; Rodríguez, Fernando; Agarwala, Richa; Landsman, David; Mariño-Ramírez, Leonardo

    2016-03-17

    Campylobacter coli is considered one of the main causes of food-borne illness worldwide. We report here the whole-genome sequence of multidrug-resistant Campylobacter coli strain COL B1-266, isolated from the Colombian poultry chain. The genome sequences encode genes for a variety of antimicrobial resistance genes, including aminoglycosides, β-lactams, lincosamides, fluoroquinolones, and tetracyclines.

  3. Simultaneous utilization of glucose, xylose and arabinose in the presence of acetate by a consortium of Escherichia coli strains

    Directory of Open Access Journals (Sweden)

    Xia Tian

    2012-06-01

    Full Text Available Abstract Background The efficient microbial utilization of lignocellulosic hydrolysates has remained challenging because this material is composed of multiple sugars and also contains growth inhibitors such as acetic acid (acetate. Using an engineered consortium of strains derived from Escherichia coli C and a synthetic medium containing acetate, glucose, xylose and arabinose, we report on both the microbial removal of acetate and the subsequent simultaneous utilization of the sugars. Results In a first stage, a strain unable to utilize glucose, xylose and arabinose (ALS1392, strain E. coli C ptsG manZ glk crr xylA araA removed 3 g/L acetate within 30 hours. In a subsequent second stage, three E. coli strains (ALS1370, ALS1371, ALS1391, which are each engineered to utilize only one sugar, together simultaneously utilized glucose, xylose and arabinose. The effect of non-metabolizable sugars on the metabolism of the target sugar was minimal. Additionally the deletions necessary to prevent the consumption of one sugar only minimally affected the consumption of a desired sugar. For example, the crr deletion necessary to prevent glucose consumption reduced xylose and arabinose utilization by less than 15% compared to the wild-type. Similarly, the araA deletion used to exclude arabinose consumption did not affect xylose- and glucose-consumption. Conclusions Despite the modest reduction in the overall rate of sugar consumption due to the various deletions that were required to generate the consortium of strains, the approach constitutes a significant improvement in any single-organism approach to utilize sugars found in lignocellulosic hydrolysate in the presence of acetate.

  4. Enumeration of sublethally injured Escherichia coli O157:H7 ATCC 43895 and Escherichia coli strain B-41560 using selective agar overlays versus commercial methods.

    Science.gov (United States)

    Smith, Amanda R; Ellison, Alysha L; Robinson, Amanda L; Drake, Maryanne; McDowell, Susan A; Mitchell, James K; Gerard, Patrick D; Heckler, Rachel A; McKillip, John L

    2013-04-01

    Quality control procedures during food processing may involve direct inoculation of food samples onto appropriate selective media for subsequent enumeration. However, sublethally injured bacteria often fail to grow, enabling them to evade detection and intervention measures and ultimately threaten the health of consumers. This study compares traditional selective and nonselective agar-based overlays versus two commercial systems (Petrifilm and Easygel) for recovery of injured E. coli B-41560 and O157:H7 strains. Bacteria were propagated in tryptic soy broth (TSB), ground beef slurry, and infant milk formula to a density of 10(6) to 10(8) CFU/ml and then were stressed for 6 min either in lactic acid (pH 4.5) or heat shocked for 3 min at 60°C. Samples were pour plated in basal layers of either tryptic soy agar (TSA), sorbitol MacConkey agar (SMAC), or violet red bile agar (VRB) and were resuscitated for 4 h prior to addition of agar overlays. Other stressed bacteria were plated directly onto Petrifilm and Easygel. Results indicate that selective and nonselective agar overlays recovered significantly higher numbers (greater than 1 log) of acid- and heat-injured E. coli O157:H7 from TSB, ground beef, and infant milk formula compared with direct plating onto selective media, Petrifilm, or Easygel, while no significant differences among these media combinations were observed for stressed E. coli B-41560. Nonstressed bacteria from TSB and ground beef were also recovered at densities significantly higher in nonselective TSA-TSA and in VRB-VRB and SMAC-SMAC compared with Petrifilm and Easygel. These data underscore the need to implement food safety measures that address sublethally injured pathogens such as E. coli O157:H7 in order to avoid underestimation of true densities for target pathogens.

  5. Identification, cloning and sequencing of Escherichia coli strain chi1378 (O78:K80) iss gene isolated from poultry colibacillosis in Iran.

    Science.gov (United States)

    Derakhshandeh, A; Zahraei Salehi, T; Tadjbakhsh, H; Karimi, V

    2009-09-01

    To identify, clone and sequence the iss (increased serum survival) gene from E. coli strain chi1378 isolated from Iranian poultry and to predict its protein product, Iss. The iss gene from E. coli strain chi1378 was amplified and cloned into the pTZ57R/T vector and sequenced. From the DNA sequence, the Iss predictive protein was evaluated using bioinformatics. Iss from strain chi1378 had 100% identity with other E. coli serotypes and isolates from different origins and also 98% identity with E. coli O157:H7 Iss protein. Phylogenetic analysis showed no significant different phylogenic groups among E. coli strains. The strong association of predicted Iss protein among different E. coli strains suggests that it could be a good antigen to control and detect avian pathogenic E. coli (APEC). Because the exact pathogenesis and the role of virulence factors are unknown, the Iss protein could be used as a target for vaccination in the future, but further research is required.

  6. Mutations by near-ultraviolet radiation in Escherichia coli strains lacking superoxide dismutase

    Energy Technology Data Exchange (ETDEWEB)

    Hoerter, J. (Stephens College, Columbia (USA). Department of Natural Sciences); Eisenstark, A. (Missouri Univ., Columbia, MO (USA). Div. of Biological Sciences Paris 7 Universite (France). Centre National de la Recherche, Institut Jacques Monod); Touati, D. (Paris 7 Universite (France). Centre National de la Recherche, Institut Jacques Monod)

    1989-12-01

    In wild-type Eschericia coli, near-ultraviolet radiation (NUV) was only weakly mutagenic. However, in an allelic mutant strain (sodA sodB) that lacks both Mn- and Fe-superoxide dismutase (SOD) and assumed to have excess superoxide anion, NUV induced a 9-fold increase in mutatin above the level that normally occurs in this double mutant. When a sodA sodB double mutant contained a plasmid carrying katG{sup +} (excess HP-I catalase), mutation by NUV was reduced to wild-type (sodA{sup +} sodB{sup +}) levels. Also, in the sodA sodB xthA triple mutant, which lacks exonuclease III (exoIII) in addition to SOD, the mutational frequency by NUV was reduced to wild-type levels. This synergistic action of NUV and O{sub 2}{sup {minus}} suggested that pre-mutational lesions occur, with exoIII converting these lesions to stable mutants. Exposure to H{sub 2}O{sub 2} induced a 2.8-fold increase in mutations in sodA sodB double mutants, but was reduced to control levels when a plasmid carrying katG{sup +} was introduced. These results suggest that NUV, in addition to its other effects on cells, increases mutations indirectly by increasing the flux of OH{sup .} radicals, possibly by generating excess H{sub 2}O{sub 2}. (author). 20 refs.; 1 fig.; 1 tab.

  7. Draft genome sequences of three Escherichia coli strains with different In Vivo pathogenicities in an avian (Ascending) infection model of the oviduct

    DEFF Research Database (Denmark)

    Olsen, Rikke Heidemann; Thøfner, Ida; Pors, Susanne Elisabeth

    2015-01-01

    Here, we present three draft genome sequences of Escherichia coli strains that experimentally were proven to possess low (strain D2-2), intermediate (Chronic_salp), or high virulence (Cp6salp3) in an avian (ascending) infection model of the oviduct.......Here, we present three draft genome sequences of Escherichia coli strains that experimentally were proven to possess low (strain D2-2), intermediate (Chronic_salp), or high virulence (Cp6salp3) in an avian (ascending) infection model of the oviduct....

  8. Characterisation of Shiga toxin-producing Escherichia coli O157 strains isolated from humans in Argentina, Australia and New Zealand

    Directory of Open Access Journals (Sweden)

    Robins-Browne Roy M

    2008-03-01

    Full Text Available Abstract Background Shiga toxin-producing Escherichia coli (STEC is an important cause of bloody diarrhoea (BD, non-bloody diarrhoea (NBD and the haemolytic uraemic syndrome (HUS. In Argentina and New Zealand, the most prevalent STEC serotype is O157:H7, which is responsible for the majority of HUS cases. In Australia, on the other hand, STEC O157:H7 is associated with a minority of HUS cases. The main aims of this study were to compare the phenotypic and genotypic characteristics of STEC O157 strains isolated between 1993 and 1996 from humans in Argentina, Australia and New Zealand, and to establish their clonal relatedness. Results Seventy-three O157 STEC strains, isolated from HUS (n = 36, BD (n = 20, NBD (n = 10, or unspecified conditions (n = 7 in Argentina, Australia and New Zealand, were analysed. The strains were confirmed to be E. coli O157 by biochemical tests and serotyping. A multiplex polymerase chain reaction (PCR was used to amplify the stx1, stx2 and rfbO157 genes and a genotyping method based on PCR-RFLP was used to determine stx1 and stx2 variants. This analysis revealed that the most frequent stx genotypes were stx2/stx2c (vh-a (91% in Argentina, stx2 (89% in New Zealand, and stx1/stx2 (30% in Australia. No stx1-postive strains were identified in Argentina or New Zealand. All strains harboured the eae gene and 72 strains produced enterohaemolysin (EHEC-Hly. The clonal relatedness of strains was investigated by phage typing and pulsed-field gel electrophoresis (PFGE. The most frequent phage types (PT identified in Argentinian, Australian, and New Zealand strains were PT49 (n = 12, PT14 (n = 9, and PT2 (n = 15, respectively. Forty-six different patterns were obtained by XbaI-PFGE; 37 strains were grouped in 10 clusters and 36 strains showed unique patterns. Most clusters could be further subdivided by BlnI-PFGE. Conclusion STEC O157 strains isolated in Argentina, Australia, and New Zealand differed from each other in terms of

  9. Strain-specific impact of the high-pathogenicity island on virulence in extra-intestinal pathogenic Escherichia coli.

    Science.gov (United States)

    Smati, Mounira; Magistro, Giuseppe; Adiba, Sandrine; Wieser, Andreas; Picard, Bertrand; Schubert, Sören; Denamur, Erick

    2017-01-01

    In order to clarify the role of the high-pathogenicity island (HPI) in the experimental virulence of Escherichia coli, we constructed different deletion mutants of the entire HPI and of three individual genes (irp2, fyuA and ybtA), encoding for three main functions within the HPI. Those mutants were constructed for three phylogroup B2 strains (536-STc127, CFT073-STc73, and NU14-STc95), representative of the main B2 subgroups causing extra-intestinal infections. Transcriptional profiles obtained for the selected HPI genes irp2, fyuA and ybtA revealed similar patterns for all strains, both under selective iron-deplete conditions and in intracellular bacterial communities in vitro, with a high expression of irp2. Deletion of irp2 and ybtA abrogated yersiniabactin production, whereas the fyuA knockout was only slightly impaired for siderophore synthesis. The experimental virulence of the strains was then tested in amoeba Dictyostelium discoideum and mouse septicaemia models. No effect of any HPI mutant was observed for the two more virulent strains 536 and CFT073. In contrast, the virulence of the less virulent NU14 strain was dramatically diminished by the complete deletion of the HPI and irp2 gene whereas a lesser reduction in virulence was observed for the fyuA and ybtA deletion mutants. The two experimental virulence models gave similar results. It appears that the role of the HPI in experimental virulence is depending on the genetic background of the strains despite similar inter-strain transcriptional patterns of HPI genes, as well as of the functional class of the studied gene. Altogether, these data indicate that the intrinsic extra-intestinal virulence in the E. coli species is multigenic, with epistatic interactions between the genes. Copyright © 2016 Elsevier GmbH. All rights reserved.

  10. Alignment-free design of highly discriminatory diagnostic primer sets for Escherichia coli O104:H4 outbreak strains.

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    Leighton Pritchard

    Full Text Available BACKGROUND: An Escherichia coli O104:H4 outbreak in Germany in summer 2011 caused 53 deaths, over 4000 individual infections across Europe, and considerable economic, social and political impact. This outbreak was the first in a position to exploit rapid, benchtop high-throughput sequencing (HTS technologies and crowdsourced data analysis early in its investigation, establishing a new paradigm for rapid response to disease threats. We describe a novel strategy for design of diagnostic PCR primers that exploited this rapid draft bacterial genome sequencing to distinguish between E. coli O104:H4 outbreak isolates and other pathogenic E. coli isolates, including the historical hæmolytic uræmic syndrome (HUSEC E. coli HUSEC041 O104:H4 strain, which possesses the same serotype as the outbreak isolates. METHODOLOGY/PRINCIPAL FINDINGS: Primers were designed using a novel alignment-free strategy against eleven draft whole genome assemblies of E. coli O104:H4 German outbreak isolates from the E. coli O104:H4 Genome Analysis Crowd-Sourcing Consortium website, and a negative sequence set containing 69 E. coli chromosome and plasmid sequences from public databases. Validation in vitro against 21 'positive' E. coli O104:H4 outbreak and 32 'negative' non-outbreak EHEC isolates indicated that individual primer sets exhibited 100% sensitivity for outbreak isolates, with false positive rates of between 9% and 22%. A minimal combination of two primers discriminated between outbreak and non-outbreak E. coli isolates with 100% sensitivity and 100% specificity. CONCLUSIONS/SIGNIFICANCE: Draft genomes of isolates of disease outbreak bacteria enable high throughput primer design and enhanced diagnostic performance in comparison to traditional molecular assays. Future outbreak investigations will be able to harness HTS rapidly to generate draft genome sequences and diagnostic primer sets, greatly facilitating epidemiology and clinical diagnostics. We expect that high

  11. Proteomic analysis reveals protein expression differences in Escherichia coli strains associated with persistent versus transient mastitis

    Science.gov (United States)

    Escherichia coli is a leading cause of bacterial mastitis in dairy cattle. Typically this infection is transient in nature, causing an infection that lasts 2-3 days. However, in a minority of cases, E. coli has been shown to cause a persistent intramammary infection. The mechanisms that allow for...

  12. The plasmid-encoded regulator activates factors conferring lysozyme resistance on enteropathogenic Escherichia coli strains.

    Science.gov (United States)

    Salinger, Nina; Kokona, Bashkim; Fairman, Robert; Okeke, Iruka N

    2009-01-01

    We demonstrate that enhanced lysozyme resistance of enteropathogenic Escherichia coli requires the plasmid-encoded regulator, Per, and is mediated by factors outside the locus for enterocyte effacement. EspC, a Per-activated serine protease autotransporter protein, conferred enhanced resistance on nonpathogenic E. coli, and a second Per-regulated, espC-independent lysozyme resistance mechanism was identified.

  13. Investigation on prevalence of Escherichia coli strains carrying virulence genes ipaH, estA, eaeA and bfpA isolated from different water sources

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    Reza Ranjbar

    2016-04-01

    Full Text Available Objective: To investigate prevalence of Escherichia coli (E. coli strains carrying virulence genes ipaH, estA, eaeA and bfpA, isolated from different water sources in Alborz Province. Methods: This study was carried out in 2014. The research included all E. coli strains isolated from different surface water sources in Alborz Province of Iran. E. coli isolates were detected and identified by standard microbiological and biochemical tests. The strains were evaluated for the presence of virulence genes ipaH, estA, eaeA and bfpA by PCR using specific primers. The PCR amplicons were visualized via electrophoresis and stained with ethidium bromide. Results: One hundred E. coli strains were isolated and included in the study. The PCR results showed that 97% of the strains harbored ipaH gene. Moreover, estA, eaeA and bfpA genes were found in 37%, 31% and 3% of the isolates. Conclusions: Our finding showed that the prevalence rates of virulence genes ipaH and estA were very high among E. coli strains isolated from different surface water sources in Alborz Province. Considering their plasmid-borne nature, the risk of transmission of these genes between other bacterial species could pose a high threat to public health.

  14. Cytolethal distending toxin producing Escherichia coli O157:H43 strain T22 represents a novel evolutionary lineage within the O157 serogroup.

    Science.gov (United States)

    Sváb, Domonkos; Bálint, Balázs; Maróti, Gergely; Tóth, István

    2016-12-01

    Enterohemorrhagic Escherichia coli (EHEC) O157:H7/NM strains are significant foodborne pathogens intensively studied, while other sero- and pathotypes of the O157 serogroup only began to receive more attention. Here we report the first genome sequence of a cytolethal distending toxin (CDT-V) producing E. coli O157:H43 strain (T22) isolated from cattle. The genome consists of a 4.9Mb chromosome assembled into three contigs and one plasmid of 82.4kb. Comparative genomic investigations conducted with the core genomes of representative E. coli strains in GenBank (n=62) confirmed the separation of T22 from the EHEC and enteropathogenic (EPEC) O157 lineages. Gene content based pangenome analysis revealed as many as 261 T22-specific coding sequences without orthologs in EDL933 EHEC O157 prototypic and two phylogenetically related commensal E. coli strains. The genome sequence revealed 10 prophage-like regions which harbor several virulence-associated genes including cdt and heat-labile enterotoxin (LT-II) encoding operons. Our results indicate that the evolutionary path of T22 is largely independent from that of EHEC and EPEC O157:H7/NM strains. Thus, the CDT-producing T22 E. coli O157:H43 strain represents a unique lineage of E. coli O157.

  15. Atypical Enteropathogenic Escherichia coli Strains form Biofilm on Abiotic Surfaces Regardless of Their Adherence Pattern on Cultured Epithelial Cells

    Science.gov (United States)

    Culler, Hebert F.; Mota, Cristiane M.; Abe, Cecilia M.; Elias, Waldir P.; Sircili, Marcelo P.; Franzolin, Marcia R.

    2014-01-01

    The aim of this study was to determine the capacity of biofilm formation of atypical enteropathogenic Escherichia coli (aEPEC) strains on abiotic and biotic surfaces. Ninety-one aEPEC strains, isolated from feces of children with diarrhea, were analyzed by the crystal violet (CV) assay on an abiotic surface after 24 h of incubation. aEPEC strains representing each HEp-2 cell type of adherence were analyzed after 24 h and 6, 12, and 18 days of incubation at 37°C on abiotic and cell surfaces by CFU/cm2 counting and confocal laser scanning microscopy (CLSM). Biofilm formation on abiotic surfaces occurred in 55 (60.4%) of the aEPEC strains. There was no significant difference in biofilm biomass formation on an abiotic versus prefixed cell surface. The biofilms could be visualized by CLSM at various developmental stages. aEPEC strains are able to form biofilm on an abiotic surface with no association with their adherence pattern on HEp-2 cells with the exception of the strains expressing UND (undetermined adherence). This study revealed the capacity of adhesion and biofilm formation by aEPEC strains on abiotic and biotic surfaces, possibly playing a role in pathogenesis, mainly in cases of persistent diarrhea. PMID:24883330

  16. Atypical Enteropathogenic Escherichia coli Strains form Biofilm on Abiotic Surfaces Regardless of Their Adherence Pattern on Cultured Epithelial Cells

    Directory of Open Access Journals (Sweden)

    Hebert F. Culler

    2014-01-01

    Full Text Available The aim of this study was to determine the capacity of biofilm formation of atypical enteropathogenic Escherichia coli (aEPEC strains on abiotic and biotic surfaces. Ninety-one aEPEC strains, isolated from feces of children with diarrhea, were analyzed by the crystal violet (CV assay on an abiotic surface after 24 h of incubation. aEPEC strains representing each HEp-2 cell type of adherence were analyzed after 24 h and 6, 12, and 18 days of incubation at 37°C on abiotic and cell surfaces by CFU/cm2 counting and confocal laser scanning microscopy (CLSM. Biofilm formation on abiotic surfaces occurred in 55 (60.4% of the aEPEC strains. There was no significant difference in biofilm biomass formation on an abiotic versus prefixed cell surface. The biofilms could be visualized by CLSM at various developmental stages. aEPEC strains are able to form biofilm on an abiotic surface with no association with their adherence pattern on HEp-2 cells with the exception of the strains expressing UND (undetermined adherence. This study revealed the capacity of adhesion and biofilm formation by aEPEC strains on abiotic and biotic surfaces, possibly playing a role in pathogenesis, mainly in cases of persistent diarrhea.

  17. Contruction of the Genetic Engineering Strain Expressed Nontoxic ST1-LTB Fusion Protein Against Enterotoxigenic Eschenichia coli

    Institute of Scientific and Technical Information of China (English)

    BAI Jia-ning; SUN Yi-min; BIAN Yan-qing; ZHAO Bao-hua

    2004-01-01

    Thermostable enterotoxinⅠ(ST1)mutant genes and thermolabile enterotoxin B subunit(LTB)genes were amplified by PCR from plasmids of Eschenichia coli C83902.The recombinant expression plasmid pZST3LTB containing ST1-LTB fusion gene was constructed by recombinant DNA technique and then transformed into Escherichia coli BL21(DE3).The ST1-LTB fusion protein was highly expressed in recombinant strain BL21(DE3)(pZST3LTB)and the fusion protein was about 38.53% of total cellular protein by SDS-PAGE and thin-layer gel scanning analysis.More important,mice immunized with crude preparation containing the fusion protein inclusion bodies or inactivated recombinant strain produced antibodies that were able to recognize ST1 in vitro.These sera antibodies were able to neutralize the biological activity of native ST1 in the suckling mouse assay.Hence the ST1-LTB fusion protein was nontoxic and immunogenic,the constructed recombinant strain BL21(DE3)(pZST3LTB)could be used as a candidate of vaccine strain.

  18. Monitoring and characterization of extended-spectrum beta-lactamases in Escherichia coli strains from healthy and sick animals in Spain in 2003.

    Science.gov (United States)

    Briñas, Laura; Moreno, Miguel Angel; Teshager, Tirushet; Sáenz, Yolanda; Porrero, María Concepción; Domínguez, Lucas; Torres, Carmen

    2005-03-01

    Genes encoding CTX-M-14, CTX-M-9, CTX-M-1, CTX-M-32, SHV-12, TEM-52, or CMY-2 beta-lactamases were detected in 21 Escherichia coli strains recovered during 2003 from sick animals (11 of 459 [2.4%] strains) and healthy animals (10 of 158 [6.3%] strains) in Spain. Twelve of these strains harbored bla(CTX-M) genes and showed unrelated pulsed-field gel electrophoresis patterns.

  19. Monitoring and Characterization of Extended-Spectrum β-Lactamases in Escherichia coli Strains from Healthy and Sick Animals in Spain in 2003

    Science.gov (United States)

    Briñas, Laura; Moreno, Miguel Angel; Teshager, Tirushet; Sáenz, Yolanda; Porrero, María Concepción; Domínguez, Lucas; Torres, Carmen

    2005-01-01

    Genes encoding CTX-M-14, CTX-M-9, CTX-M-1, CTX-M-32, SHV-12, TEM-52, or CMY-2 β-lactamases were detected in 21 Escherichia coli strains recovered during 2003 from sick animals (11 of 459 [2.4%] strains) and healthy animals (10 of 158 [6.3%] strains) in Spain. Twelve of these strains harbored blaCTX-M genes and showed unrelated pulsed-field gel electrophoresis patterns. PMID:15728945

  20. [Surveillance of Antimicrobial Resistant Esherichia coli by Rectal Swab Method--Annual Change of Prevalence of Quinolone-resistant and ESBL Producing Strains from 2009 to 2013].

    Science.gov (United States)

    Nasu, Yoshitsugu; Sako, Shinichi; Yano, Tomofumi; Kosaka, Noriko

    2015-09-01

    Although most of commonly used antimicrobial agents had been susceptible to Esherichia coli, recently there are a lot of reports concerning about community-acquired infection caused by resistant E. coli. The aim of this study is to define the prevalence of resistant E. coli in normal flora colonization by the rectal swab method. From June 2009 to December 2013, 251 male patients (50-85 year-old, median 68) planned to transrectal prostate biopsy participated in this study. Stools stuck on the glove at the digital examination were provided for culture specimen. Identification of E. coli and determination of MIC was performed by MicroScan WalkAway40plus (Siemens). Isolated E. coli were deemed quinolone-resistant strains when their MIC of levofloxacine was 4 μg/mL or above according to the breakpoint MIC by the CLSI criteria. ESBL producing ability was determined by the double disk method used by CVA contained ESBL definition disc (Eikenkagaku). Of the 251 study patients, 224 patients had positive cultures of E. coli. Twenty-four patients had quinolone-resistant strains and 9 patients had ESBL producing strains. The prevalence of quinolone-resistant strains in 2009, 2010, 2011, 2012 and 2013 were 5.9% (2 out of 34 strains), 13.5% (5 out of 37 strains), 12.5% (4 out of 32 strains), 9.0% (6 out of 67) and 13.0% (7 out of 54 strains), respectively. The prevalence of ESBL producing strains in 2009, 2010, 2011, 2012 and 2013 were 0% (0 out of 34 strains), 5.4% (2 out of 37 strains), 3.1% (1 out of 32 strains), 3.0% (2 out of 67 strains) and 7.4% (4 out of 54 strains), respectively. In 2013, the prevalence of antimicrobial resistant E. coli, both quinolone-resistant and ESBL producing strains, were increasing. We have to pay a close attention to the increase of resistant E. coli.

  1. Biosorption of lead by e. coli strains expressingvitreoscilla hemoglobin: isotherm modeling with two-and three-parameter models

    Energy Technology Data Exchange (ETDEWEB)

    Aljundi, Isam H. [Chemical Engineering Department, Mutah University, Al-Karak (Jordan); Khleifat, Khaled M. [Biology Department, Mutah University, Al-Karak (Jordan)

    2010-06-15

    Biosorption is presented as an alternative choice to traditional physicochemical means for removing toxic metals from groundwater and wastewaters. Removal of lead (Pb) from solutions was studied using Escherichia coli (parental) and Vitreoscilla hemoglobin (VHb)-expressing E. coli (transformed) cells. Pb biosorption was increased in bacterial hemoglobin-expressing E. coli cells grown in Luria broth B containing different concentrations of Pb{sup 2+}. The maximum Pb{sup 2+} biosorption of transformed and parental cells was determined to be 612 and 370 {mu}g Pb/g biomass, respectively. The inhibitory effect of Pb{sup 2+} on the parental strain was determined at 10 ppm. However, in transformed cells, Pb{sup 2+} was lethal at 100 ppm. The optimum aeration required for the transformed cells was lower than that for the parental strain on a growth yield basis. A linear correlation was established between the biosorption and uptake amounts. The biosorption process was analyzed using two-parameter (Langmuir, Freundlich, Temkin) and three-parameter models (Sips, Redlich-Peterson, Toth). The chi-square test was used to compare these models. It was demonstrated that the three-parameter model is better in describing biosorption equilibria a, white the Sips equation showed the best description for both types of cells. (Abstract Copyright [2010], Wiley Periodicals, Inc.)

  2. Antibiotic resistance patterns of Escherichia coli strains isolated from surface water and groundwater samples in a pig production area

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    Roger Neto Schneider

    2009-09-01

    Full Text Available The use of antibiotics, so excessive and indiscriminate in intensive animal production, has triggered an increase in the number of resistant microorganisms which can be transported to aquatic environments. The aim of this study was to determine the profile of the antimicrobial resistance of samples of Escherichia coli isolated from groundwater and surface water in a region of pig breeding. Through the test of antimicrobial susceptibility, we analyzed 205 strains of E. coli. A high rate of resistance to cefaclor was observed, both in surface water (51.9% and groundwater (62.9%, while all samples were sensitive to amikacin. The percentages of multi-resistant samples were 25.96% and 26.73% in surface water and groundwater, respectively, while 19.23% and 13.86% were sensitive to all antibiotics tested. It was determined that the rate of multiple antibiotic resistance (MAR was 0.164 for surface water and 0.184 for groundwater. No significant differences were found in the profile of the antimicrobial resistance in strains of E. coli isolated in surface water and groundwater, but the index MAR calculated in certain points of groundwater may offer a potential risk of transmission of resistant genes.

  3. Estimation of transmission parameters of a fluoroquinolone-resistant Escherichia coli strain between pigs in experimental conditions

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    Andraud Mathieu

    2011-03-01

    Full Text Available Abstract Antimicrobial resistance is of primary importance regarding public and animal health issues. Persistence and spread of resistant strains within a population contribute to the maintenance of a reservoir and lead to treatment failure. An experimental trial was carried out to study the horizontal transmission of a fluoroquinolone-resistant Escherichia coli strain from inoculated to naïve pigs. All naïve contact pigs had positive counts of fluoroquinolone-resistant E. coli after only two days of contact. Moreover, re-infections of inoculated pigs caused by newly contaminated animals were suspected. A maximum likelihood method, based on a susceptible-infectious-susceptible (SIS model, was used to determine the transmission parameters. Two transmission levels were identified depending on the quantity of bacteria shed by infected individuals: (i low-shedders with bacterial counts of resistant E. coli in the faeces between 5*103 and 106 CFU/g (βL = 0.41 [0.27; 0.62], (ii high shedders with bacterial counts above 106 CFU/g (βH = 0.98 [0.59; 1.62]. Hence, transmission between animals could be pivotal in explaining the persistence of resistant bacteria within pig herds.

  4. Oxidative phosphorylation in Escherichia coli. Characterization of mutant strains in which F1-ATPase contains abnormal beta-subunits.

    Science.gov (United States)

    Senior, A E; Langman, L; Cox, G B; Gibson, F

    1983-02-15

    To facilitate study of the role of the beta-subunit in the membrane-bound proton-translocating ATPase of Escherichia coli, we identified mutant strains from which an F1-ATPase containing abnormal beta-subunits can be purified. Seventeen strains of E. coli, characterized by genetic complementation tests as carrying mutations in the uncD gene (which codes for the beta-subunit), were studied. The majority of these strains (11) were judged to be not useful, as their membranes lacked ATPase activity, and were either proton-permeable as prepared or remained proton-impermeable after washing with buffer of low ionic strength. A further two strains were of a type not hitherto reported, in that their membranes had ATPase activity, were proton-impermeable as prepared, and were not rendered proton-permeable by washing in buffer of low ionic strength. Presumably in these two strains F1-ATPase is not released in soluble form by this procedure. F1-ATPase of normal molecular size were purified from strains AN1340 (uncD478), AN937 (uncD430), AN938 (uncD431) and AN1543 (uncD484). F1-ATPase from strain AN1340 (uncD478) had 15% of normal specific Mg-dependent ATPase activity and 22% of normal ATP-synthesis activity. The F1-ATPase preparations from strains AN937, AN938 and AN1543 had respectively 1.7%, 1.8% and 0.2% of normal specific Mg-dependent ATPase activity, and each of these preparations had very low ATP-synthesis activity. The yield of F1-ATPase from the four strains described was almost twice that obtained from a normal haploid strain. The kinetics of Ca-dependent ATPase activity were unusual in each of the four F1-ATPase preparations. It is likely that these four mutant uncD F1-ATPase preparations will prove valuable for further experimental study of the F1-ATPase catalytic mechanism.

  5. Virulence factors in Escherichia coli strains isolated from urinary tract infection and pyometra cases and from feces of healthy dogs.

    Science.gov (United States)

    Siqueira, Amanda K; Ribeiro, Marcio G; Leite, Domingos da S; Tiba, Monique R; Moura, Claudia de; Lopes, Maria Denise; Prestes, Nereu Carlos; Salerno, Tatiana; Silva, Aristeu V da

    2009-04-01

    The aim of this study was to compare the prevalence of virulence genes in 158 Escherichia coli strains isolated from 51 clinical cases of UTIs, 52 of pyometra and from 55 fecal samples from healthy dogs by PCR. papC was found in 12 (23.5%) strains isolated from UTIs, 19 (36.5%) from pyometra and 10 (18.2%) from feces. papGII was observed in 3 (5.8%) strains from pyometra, and papGIII in 10 (19.6%) from UTIs, 15 (28.8%) from pyometra and 9 (16.4%) from feces. sfaS was detected in 22 (43.1%) strains from UTIs, 24 (46.1%) from pyometra and 19 (34.5%) from feces. hlyA was observed in 17 (33.3%) strains from UTIs, 18 (34.6%) from pyometra and 7 (12.7%) from feces, while cnf-1 was detected in 11 (21.6%) from UTIs, 21 (40.4%) from pyometra and 9 (16.4%) from feces. iucD was observed in 12 (23.5%) strains from UTIs, 9 (17.3%) from pyometra and 1 (1.8%) from feces. usp was found 17 (33.3%) isolates from UTIs and 36 (69.9%) from pyometra.

  6. Short communication: The role of autoinducer 2 (AI-2) on antibiotic resistance regulation in an Escherichia coli strain isolated from a dairy cow with mastitis.

    Science.gov (United States)

    Xue, Ting; Yu, Lumin; Shang, Fei; Li, Wenchang; Zhang, Ming; Ni, Jingtian; Chen, Xiaolin

    2016-06-01

    Extended spectrum β-lactamase (ESBL)-positive Escherichia coli is a major etiological organism responsible for bovine mastitis. The autoinducer 2 (AI-2) quorum sensing system is widely present in many species of gram-negative and gram-positive bacteria and has been proposed to be involved in interspecies communication. In E. coli model strains, the functional mechanisms of AI-2 have been well studied; however, in clinical antibiotic-resistant E. coli strains, whether AI-2 affects the expression of antibiotic resistance genes has not been reported. In this study, we report that exogenous AI-2 increased the antibiotic resistance of a clinical E. coli strain isolated from a dairy cow with mastitis by upregulating the expression of TEM-type enzyme in an LsrR (LuxS regulated repressor)-dependent manner.

  7. High-Level Genotypic Variation and Antibiotic Sensitivity among Escherichia coli O157 Strains Isolated from Two Scottish Beef Cattle Farms

    Science.gov (United States)

    Vali, Leila; Wisely, Karen A.; Pearce, Michael C.; Turner, Esther J.; Knight, Hazel I.; Smith, Alastair W.; Amyes, Sebastian G. B.

    2004-01-01

    Escherichia coli O157:H7 is a human pathogen that is carried and transmitted by cattle. Scotland is known to have one of the highest rates of E. coli O157 human infections in the world. Two hundred ninety-three isolates were obtained from naturally infected cattle and the environment on two farms in the Scottish Highlands. The isolates were typed by pulsed-field gel electrophoresis (PFGE) with XbaI restriction endonuclease enzyme, and 19 different variations in patterns were found. There was considerable genomic diversity within the E. coli O157 population on the two farms. The PFGE pattern of one of the observed subtypes matched exactly with that of a strain obtained from a Scottish patient with hemolytic-uremic syndrome. To examine the stability of an individual E. coli O157 strain, continuous subculturing of a strain was performed 110 times. No variation from the original PFGE pattern was observed. We found three indistinguishable subtypes of E. coli O157 on both study farms, suggesting common sources of infection. We also examined the antibiotic resistance of the isolated strains. Phenotypic studies demonstrated resistance of the strains to sulfamethoxazole (100%), chloramphenicol (3.07%), and at a lower rate, other antibiotics, indicating the preservation of antibiotic sensitivity in a rapidly changing population of E. coli O157. PMID:15466537

  8. Recovery of electric energy from formate by using a recombinant strain of Escherichia coli.

    Science.gov (United States)

    Ojima, Yoshihiro; Kawata, Teruyoshi; Matsuo, Nahoko; Nishinoue, Yosuke; Taya, Masahito

    2014-10-01

    Recombinant Escherichia coli cells were applied for the recovery of electric energy from formate. Initially, the fdh gene, which encodes formate dehydrogenase (FDH) of Mycobacterium vaccae, was introduced into E. coli cells to allow efficient degradation of formate. The constructed microbial fuel cell (MFC) with E. coli BW25113 cells carrying fdh gene showed appreciable generation of current density in the presence of formate as a substrate. Current density and polarization curves revealed that the performance of MFC under examined conditions was limited by the electron transfer from bulk liquid to the electrode surface; accordingly, agitation resulted in an increase in the current density and achieved a coulombic efficiency of 21.7 % on the basis of formate consumed. Thus, gene recombination enables E. coli cells to utilize formate as a fuel for MFC.

  9. Expression of curli by Escherichia coli O157:H7 strains isolated from patients during outbreaks is different from similar strains isolated from leafy green production environments

    Directory of Open Access Journals (Sweden)

    Subbarao Venkata Ravva

    2016-12-01

    Full Text Available We previously reported that the strains of Escherichia coli O157:H7 (EcO157 that survived longer in austere soil environment lacked expression of curli, a fitness trait linked with intestinal colonization. In addition, the proportion of curli-positive variants of EcO157 decreased with repeated soil exposure. Here we evaluated 84 and 176 clinical strains from outbreaks and sporadic infections in the US, plus 211 animal fecal and environmental strains for curli expression. These shiga-toxigenic strains were from 328 different genotypes, as characterized by multi-locus variable-number tandem-repeat analysis (MLVA. More than half of the fecal strains (human and animal and a significant proportion of environmental isolates (82% were found to lack curli expression. EcO157 strains from several outbreaks linked with the consumption of contaminated apple juice, produce, hamburgers, steak and beef were also found to lack curli expression. Phylogenetic analysis of fecal strains indicates curli expression is distributed throughout the population. However, a significant proportion of animal fecal isolates (84% gave no curli expression compared to human fecal isolates (58%. In addition, analysis of environmental isolates indicated nearly exclusive clustering of curli expression to a single branch of the minimal spanning tree. This indicates that curli expression depends primarily upon the type of environmental exposure and the isolation source, although genotypic differences also contribute to clonal variation in curli. Furthermore, curli-deficient phenotype appears to be a selective trait for survival of EcO157 in agricultural environments.

  10. A study of the surface charging properties of a standard strain of Escherichia coli (ATCC 11775) in aqueous solutions.

    Science.gov (United States)

    Li, Xinpeng; Xue, Xinkai; Pashley, Richard M

    2015-11-01

    In this study, the Escherichia coli (E. coli) strain ATCC 11775 was studied to determine its surface charging properties in a range of different aqueous salt solutions, with the aim of evaluating its potential as a monitor organism for water treatment. Zeta potential measurements were carried out in various solutions containing: NaCl, CaCl2, MgSO4, ZnSO4 and C14TAB, at different pH values and concentrations. Interestingly, it was found that the zeta potential of this strain of E. coli remained fairly constant at pH values over about 6, in 1mM NaCl solutions. In order to explain the cell surface charging properties, a simple, mass action surface ionization model was developed. This model indicates that the surface charging of these E. coli cells can be modeled simply using the ionization behavior of the acid groups in the common anionic membrane lipid phosphatidylserine (PS). There appeared to be no specific, strong adsorption of either divalent anions or cations, until high salt concentrations, above about 0.1M. The results suggest that at high concentrations both Ca(2+) and SO4(2-) ions are strongly adsorbed at the cell surface. However reduction of the magnitude of the surface electrostatic potential, due to Ca(2+) ion adsorption, did not appear to cause any cellular binding. In comparison, cationic surfactant was strongly adsorbed by the cell membrane surface, even at concentrations of 0.1mM, and light scattering studies indicated that the adsorption of the surfactant appeared to lyse the cell membrane and release internal cellular materials leading to a significant reduction in cell size.

  11. Epidemiology of Antimicrobial Resistance among Escherichia coli Strains in Trans-Nzoia County, Kenya

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    Collins Kibenei Kipkorir, Philip Bett, Patrick O. Onyango

    2016-09-01

    Full Text Available Objective: This study aimed to determine prevalence, antimicrobial susceptibility profile and the genetic basis to antimicrobial resistance, targeting blaTEM gene expression of diarrheagenic Escherichia coli among patients suffering from gastroenteritis in Kitale County Referral Hospital. Methods: A cross-sectional study design was adopted. A total of 103 fecal specimens were collected from participants ranging in age from two weeks to 82 years. E. coli was isolated and identified based on phenotypic and biochemical properties. Antimicrobial susceptibility was determined by Kirby-Bauer disk diffusion method. Polymerase chain reaction was used to detect the presence of blaTEM gene. Results: The prevalence of E. coli was 90.2% and age of the patient explained 53% of variation in prevalence. Isolates of diarrheagenic E. coli showed varied degree of susceptibility with sulfamethoxazole at 97%, co-trimoxazole 96%, ampicillin 84%, chloramphenicol 27%, tetracycline 16%, kanamycin 10% and streptomycin 9%. However, E. coli was highly sensitive to gentamicin at 96.8%. Approximately 42.2% of E. coli isolates were multidrug resistant to sulfamethoxazole, co-trimoxazole, ampicillin, chloramphenicol, tetracycline, kanamycin and streptomycin. All isolates that were resistant to ampicillin harbored blaTEM gene suggesting genetic mediation. Conclusion: The observed pattern of resistance to antibiotics points to the need to regulate their use and arrest buildup of resistant genes within the population. J Microbiol Infect Dis 2016;6(3: 107-112

  12. Effect of different feed ingredients and additives on IPEC-J2 cells challenged with an enterotoxigenic Escherichia coli strain.

    Science.gov (United States)

    Spitzer, F; Speiser, S; Vahjen, W; Zentek, J

    2016-08-01

    The intestinal porcine epithelial cell line IPEC-J2 was used as an in vitro model to assess effects of additives on the adhesion and cell toxic effects of a F4-positive (ETEC) and a F4-negative Escherichia coli (DSM 2840) strain. Bacterial adhesion was examined using flow cytometry in IPEC-J2 cells infected with bacteria stained with 5,6-carboxymethyl fluorescein diacetate succinimidyl ester. Measurement of transepithelial electrical resistance (TEER) was performed to characterize the impact on IPEC-J2 monolayer integrity. The feed additives were prepared as aqueous extract and tested in different dilutions and incubation times. The F4-positive ETEC strain had a high adhesion to IPEC-J2 cells and reduced TEER shortly after the in vitro infection. The nonpathogenic E. coli strain DSM 2840 showed only low adhesion capacity and no TEER impairment. Infection with ETEC with added test extracts showed a reduction of bacterial adhesion to IPEC-J2 cells by an autolyzed yeast product (p < 0.05). Bovine colostrum, an additive containing thyme extract and an organic acid mix did not interfere with the ETEC adherence. The TEER decrease of the IPEC-J2 monolayer after ETEC infection was not affected by the added substances. In conclusion, interference with epithelial adhesion might be a protective mechanism of the tested yeast extract, indicating that the cell culture model might be suitable as screening tool to complement in vivo challenge trials with piglets.

  13. Influence of Apple Cultivars on Inactivation of Different Strains of Escherichia coli O157:H7 in Apple Cider by UV Irradiation

    Science.gov (United States)

    Basaran, N.; Quintero-Ramos, A.; Moake, M. M.; Churey, J. J.; Worobo, R. W.

    2004-01-01

    This study examined the effect of different apple cultivars upon the UV inactivation of Escherichia coli O157:H7 strains within unfiltered apple cider. Apple cider was prepared from eight different apple cultivars, inoculated with approximately 106 to 107 CFU of three strains of E. coli O157:H7 per ml (933, ATCC 43889, and ATCC 43895), and exposed to 14 mJ of UV irradiation per cm2. Bacterial populations for treated and untreated samples were then enumerated by using nonselective media. E. coli O157:H7 ATCC 43889 showed the most sensitivity to this disinfection process with an average 6.63-log reduction compared to an average log reduction of 5.93 for both strains 933 and ATCC 43895. The highest log reduction seen, 7.19, occurred for strain ATCC 43889 in Rome cider. The same cider produced the lowest log reductions: 5.33 and 5.25 for strains 933 and ATCC 43895, respectively. Among the apple cultivars, an average log reduction range of 5.78 (Red Delicious) to 6.74 (Empire) was observed, with two statistically significant (α ≤ 0.05) log reduction groups represented. Within the paired cultivar-strain analysis, five of eight ciders showed statistically significant (α ≤ 0.05) differences in at least two of the E. coli strains used. Comparison of log reductions among the E. coli strains to the cider parameters of °Brix, pH, and malic acid content failed to show any statistically significant relationship (R2 ≥ 0.95). However, the results of this study indicate that regardless of the apple cultivar used, a minimum 5-log reduction is achieved for all of the strains of E. coli O157:H7 tested. PMID:15466551

  14. [Resistant phenotypes of Escherichia coli strains responsible for urinary tract infection in the laboratory of the University Hospital Joseph Raseta Befelatanana, Antananarivo].

    Science.gov (United States)

    Rakotovao-Ravahatra, Zafindrasoa Domoina; Randriatsarafara, Fidiniaina Mamy; Rasoanandrasana, Saïda; Raverohanta, Léa; Rakotovao, Andriamiadana Luc

    2017-01-01

    Urinary tract infection caused by Escherichia coli frequently occurs in the hospital environment. This study aims to describe resistant phenotypes of Escherichia coli strains to monitor their occurrence. We conducted a descriptive retrospective study of 102 Escherchia coli strains responsible for urinary tract infection in the laboratory of the University Hospital Joseph Raseta Befelatanana, Antananarivo from January 2014 to October 2016. Beta-lactam antibiotic resistance screening identified high-level penicillinases 50% (n=51), Escherichia coli producing extended-spectrum beta-lactamases (ESBLs) 22.5% (n=23), high-level cephalosporinases 14.7% (n=15), penicillinases low level 5.9% (n=6), wild type strains 5.9% (n=6) and a strain ofEscherichia coli emerging strain high-level resistance. Aminoglycosides resistance was identified in 58 (56.9%) wild type phenotype, 29 (28.4%) strains sensitive to amikacin and 15 (14.7%) resistant to all aminoglycosides. Fluoroquinolones resistance was identified in 52 (51%) wild type strains, 9 (8.8%) strains sensitive to ciprofloxacin and 41 (40.2%) resistant to all fluoroquinolones. Women (25, 7%) (p= 0.25, NS), patients more than 60 years (38.7%) (p=0.02), patients hospitalized in the Department of Nephrology (53.8%) (p=0.04), with urinary disorder and kidney disease (29, 7%) (p= 0.2, NS) were the most affected by E-ESBL. Based on high multidrug resistance in Escherichia coli strains guidelines for the empirical treatment of urinary tract infections need to be revised.

  15. Investigation of environmental factors on the prevalence of free bacteriophages against Shiga toxin-producing Escherichia coli strains in produce pre-harvest environment in Salinas, California

    Science.gov (United States)

    Shiga toxin-producing E. coli (STEC) strains, commensal to gastrointestinal tracts of ruminants or other animals, have been associated with serious human illnesses and high mortality among immunocompromised populations. Along with the detection of STEC strains from fecal-contaminated environments su...

  16. Genetic Characterization and Immunogenicity of Coli Surface Antigen 4 from Enterotoxigenic Escherichia coli when It Is Expressed in a Shigella Live-Vector Strain

    Science.gov (United States)

    Altboum, Zeev; Levine, Myron M.; Galen, James E.; Barry, Eileen M.

    2003-01-01

    The genes that encode the enterotoxigenic Escherichia coli (ETEC) CS4 fimbriae, csaA, -B, -C, -E, and -D′, were isolated from strain E11881A. The csa operon encodes a 17-kDa major fimbrial subunit (CsaB), a 40-kDa tip-associated protein (CsaE), a 27-kDa chaperone-like protein (CsaA), a 97-kDa usher-like protein (CsaC), and a deleted regulatory protein (CsaD′). The predicted amino acid sequences of the CS4 proteins are highly homologous to structural and assembly proteins of other ETEC fimbriae, including CS1 and CS2, and to CFA/I in particular. The csaA, -B, -C, -E operon was cloned on a stabilized plasmid downstream from an osomotically regulated ompC promoter. pGA2-CS4 directs production of CS4 fimbriae in both E. coli DH5α and Shigella flexneri 2a vaccine strain CVD 1204, as detected by Western blot analysis and bacterial agglutination with anti-CS4 immune sera. Electron-microscopic examination of Shigella expressing CS4 confirmed the presence of fimbriae on the bacterial surface. Guinea pigs immunized with CVD 1204(pGA2-CS4) showed serum and mucosal antibody responses to both the Shigella vector and the ETEC fimbria CS4. Among the seven most prevalent fimbrial antigens of human ETEC, CS4 is the last to be cloned and sequenced. These findings pave the way for CS4 to be included in multivalent ETEC vaccines, including an attenuated Shigella live-vector-based ETEC vaccine. PMID:12595452

  17. Growth inhibition of Staphylococcus aureus and escherichia coli strains by neutralizing IgY antibodies from ostrich egg yolk

    Directory of Open Access Journals (Sweden)

    Fernando Luiz Tobias

    2012-06-01

    Full Text Available Ostrich raising around the world have some key factors and farming profit depend largely on information and ability of farmers to rear these animals. Non fertilized eggs from ostriches are discharged in the reproduction season. Staphylococcus aureus and Escherichia coli are microorganisms involved in animal and human diseases. In order to optimize the use of sub products of ostrich raising, non fertilized eggs of four selected birds were utilized for development of polyclonal IgY antibodies. The birds were immunized (200ug/animal with purified recombinant staphylococcal enterotoxin C (recSEC and synthetic recRAP, both derived from S. aureus, and recBFPA and recEspB involved in E. coli pathogenicity, diluted in FCA injected in the braquial muscle. Two subsequent immunization steps with 21 days intervals were repeated in 0,85% saline in FIA. Blood and eggs samples were collected before and after immunization steps. Egg yolk immunoglobulins were purified by precipitation with 19% sodium sulfate and 20% ammonium sulphate methodologies. Purified IgY 50µL aliquots were incubated in 850µL BHI broth containing 50µL inoculums of five strains of S. aureus and five strains of E.coli during four hours at 37ºC. Growth inhibition was evaluated followed by photometry reading (DO550nm. Egg yolk IgY preparation from hiperimmunized birds contained antibodies that inhibited significantly (p<0,05 growth of strains tested. Potential use of ostrich IgY polyclonal antibodies as a diagnostic and therapeutic tool is proposed for diseased animals.

  18. Prevalence of virulence genes associated with pathogenic Escherichia coli strains isolated from domestically harvested rainwater during low- and high-rainfall periods.

    Science.gov (United States)

    Dobrowsky, P H; van Deventer, A; De Kwaadsteniet, M; Ndlovu, T; Khan, S; Cloete, T E; Khan, W

    2014-03-01

    The possible health risks associated with the consumption of harvested rainwater remains one of the major obstacles hampering its large-scale implementation in water limited countries such as South Africa. Rainwater tank samples collected on eight occasions during the low- and high-rainfall periods (March to August 2012) in Kleinmond, South Africa, were monitored for the presence of virulence genes associated with Escherichia coli. The identity of presumptive E. coli isolates in rainwater samples collected from 10 domestic rainwater harvesting (DRWH) tanks throughout the sampling period was confirmed through universal 16S rRNA PCR with subsequent sequencing and phylogenetic analysis. Species-specific primers were also used to routinely screen for the virulent genes, aggR, stx, eae, and ipaH found in enteroaggregative E. coli (EAEC), enterohemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), and enteroinvasive E. coli, respectively, in the rainwater samples. Of the 92 E. coli strains isolated from the rainwater using culture based techniques, 6% were presumptively positively identified as E. coli O157:H7 using 16S rRNA. Furthermore, virulent pathogenic E. coli genes were detected in 3% (EPEC and EHEC) and 16% (EAEC) of the 80 rainwater samples collected during the sampling period from the 10 DRWH tanks. This study thus contributes valuable information to the limited data available regarding the ongoing prevalence of virulent pathotypes of E. coli in harvested rainwater during a longitudinal study in a high-population-density, periurban setting.

  19. Functional complementation of Leishmania (Leishmania) amazonensis AP endonuclease gene (lamap) in Escherichia coli mutant strains challenged with DNA damage agents.

    Science.gov (United States)

    Verissimo-Villela, Erika; Kitahara-Oliveira, Milene Yoko; Reis, Ana Beatriz de Bragança Dos; Albano, Rodolpho Mattos; Da-Cruz, Alda Maria; Bello, Alexandre Ribeiro

    2016-05-01

    During its life cycle Leishmania spp. face several stress conditions that can cause DNA damages. Base Excision Repair plays an important role in DNA maintenance and it is one of the most conserved mechanisms in all living organisms. DNA repair in trypanosomatids has been reported only for Old World Leishmania species. Here the AP endonuclease from Leishmania (L.) amazonensis was cloned, expressed in Escherichia coli mutants defective on the DNA repair machinery, that were submitted to different stress conditions, showing ability to survive in comparison to the triple null mutant parental strain BW535. Phylogenetic and multiple sequence analyses also confirmed that LAMAP belongs to the AP endonuclease class of proteins.

  20. Functional complementation of Leishmania (Leishmania) amazonensis AP endonuclease gene (lamap) in Escherichia coli mutant strains challenged with DNA damage agents

    Science.gov (United States)

    Verissimo-Villela, Erika; Kitahara-Oliveira, Milene Yoko; dos Reis, Ana Beatriz de Bragança; Albano, Rodolpho Mattos; Da-Cruz, Alda Maria; Bello, Alexandre Ribeiro

    2016-01-01

    During its life cycle Leishmania spp. face several stress conditions that can cause DNA damages. Base Excision Repair plays an important role in DNA maintenance and it is one of the most conserved mechanisms in all living organisms. DNA repair in trypanosomatids has been reported only for Old World Leishmania species. Here the AP endonuclease from Leishmania (L.) amazonensis was cloned, expressed in Escherichia coli mutants defective on the DNA repair machinery, that were submitted to different stress conditions, showing ability to survive in comparison to the triple null mutant parental strain BW535. Phylogenetic and multiple sequence analyses also confirmed that LAMAP belongs to the AP endonuclease class of proteins. PMID:27223868

  1. In vitro testing of the some antibiotics efficiency on some E. coli strains isolated from avian farms

    Directory of Open Access Journals (Sweden)

    Veturia-Ileana Nueleanu,

    2008-06-01

    Full Text Available For broiler chickens, a major mortality cause on the entire exploitation period is represented by the collibacilar infections, which need a correct and efficient treatment. The mortality in 94 broiler chickens was studied, between three and thirty days of age, from four avian farms from Satu-Mare county, isolating and identifying 32 Escherichia coli strains in pure culture. The testing of sensitivity to antibiotics of the isolated germs was made through the difusimethric method, revealing a decreased efficiency to Oxitetracyclin, Amoxicyllin and Erithromycin, while in the case of Amoxicyllin + Clavulanic acid, Florfenicol and Gentamycin a good sensitivity was observed.

  2. Strain differences in fitness of Escherichia coli O157:H7 to resist protozoan predation and survival in soil.

    Science.gov (United States)

    Ravva, Subbarao V; Sarreal, Chester Z; Mandrell, Robert E

    2014-01-01

    Escherichia coli O157:H7 (EcO157) associated with the 2006 spinach outbreak appears to have persisted as the organism was isolated, three months after the outbreak, from environmental samples in the produce production areas of the central coast of California. Survival in harsh environments may be linked to the inherent fitness characteristics of EcO157. This study evaluated the comparative fitness of outbreak-related clinical and environmental strains to resist protozoan predation and survive in soil from a spinach field in the general vicinity of isolation of strains genetically indistinguishable from the 2006 outbreak strains. Environmental strains from soil and feral pig feces survived longer (11 to 35 days for 90% decreases, D-value) with Vorticella microstoma and Colpoda aspera, isolated previously from dairy wastewater; these D-values correlated (PVorticella (rs = -0.465; P = 0.05) or Colpoda (rs = -0.750; P = 0.0001). In contrast, protozoan growth correlated positively with C+ phenotype (Vorticella, rs = 0.730, P = 0.0004; Colpoda, rs = 0.625, P = 0.006) suggesting a preference for consumption of C+ cells, although they grew on C- strains also. We speculate that the C- phenotype is a selective trait for survival and possibly transport of the pathogen in soil and water environments.

  3. Impact of metal ion homeostasis of genetically modified Escherichia coli Nissle 1917 and K12 (W3110) strains on colonization properties in the murine intestinal tract.

    Science.gov (United States)

    Kupz, Andreas; Fischer, André; Nies, Dietrich H; Grass, Gregor; Göbel, Ulf B; Bereswill, Stefan; Heimesaat, Markus M

    2013-09-01

    Metal ions are integral parts of pro- as well as eukaryotic cell homeostasis. Escherichia coli proved a valuable in vitro model organism to elucidate essential mechanisms involved in uptake, storage, and export of metal ions. Given that E. coli Nissle 1917 is able to overcome murine colonization resistance, we generated several E. coli Nissle 1917 mutants with defects in zinc, iron, copper, nickel, manganese homeostasis and performed a comprehensive survey of the impact of metal ion transport and homeostasis for E. coli colonization capacities within the murine intestinal tract. Seven days following peroral infection of conventional mice with E. coli Nissle 1917 strains exhibiting defined defects in zinc or iron uptake, the respective mutant and parental strains could be cultured at comparable, but low levels from the colonic lumen. We next reassociated gnotobiotic mice in which the microbiota responsible for colonization resistance was abrogated by broad-spectrum antibiotics with six different E. coli K12 (W3110) mutants. Seven days following peroral challenge, each mutant and parental strain stably colonized duodenum, ileum, and colon at comparable levels. Taken together, defects in zinc, iron, copper, nickel, and manganese homeostasis do not compromise colonization capacities of E. coli in the murine intestinal tract.

  4. Pulsed-field gel electrophoresis typing of Eschericia coli strains from samples collected before and after pivmecillinam or placebo treatment of uncomplicated community-acquired urinary tract infection in women

    DEFF Research Database (Denmark)

    Ejrnæs, K; Sandvang, D; Lundgren, Bettina;

    2006-01-01

    The primary infecting Escherichia coli strains from 156 women with community-acquired uncomplicated urinary tract infection (UTI) randomized to pivmecillinam or placebo and the E. coli strains causing UTI at two follow-up visits were typed using pulsed-field gel electrophoresis (PFGE). In the piv......, constituting a reservoir for recurrent UTI.......The primary infecting Escherichia coli strains from 156 women with community-acquired uncomplicated urinary tract infection (UTI) randomized to pivmecillinam or placebo and the E. coli strains causing UTI at two follow-up visits were typed using pulsed-field gel electrophoresis (PFGE......). The finding that the majority of UTIs at follow-up are caused by the primary infecting E. coli strain supports the theory of a vaginal and rectal reservoir but could also support the recent discovery that E. coli strains are able to persist in the bladder epithelium despite appropriate antibiotic treatment...

  5. Altered alpha subunits in phenylalanyl-tRNA synthetases from p-fluorophenylalanine-resistant strains of Escherichis coli.

    Science.gov (United States)

    Hennecke, H; Böck, A

    1975-07-01

    Three different phenylalanyl-tRNA synthetases have been purified to near homogeneity, one from a wild-type strain of Escherichia coli and the others from two independently isolated p-fluorophenyalanine-resistant strains. The mutant enzymes were not able to use p-fluorophenylalanine as a substrate for activation and attachment to tRNA. They proved to be indistinguishable from the wild-type enzyme by several electrophoretic and immunological criteria. The alpha and beta subunits of all three enzymes have been prepared by a method described in this paper. The isolated subunits per se did not reveal any significant enzyme activity, but combined they were able to form active phenylalanyl tRNA synthetase after a defined reconstitution process. Mixed reconstitution experiments between wild-type and mutant subunits indicate that the mutant alpha subunit is responsible for p-fluorophenylalanine resistance and therefore seems to carry the phenylalanine-binding site or to participate in its formation.

  6. Comparative Evaluation of the Modulation of Antibiotic-Activity against Strains of Escherichia coli and Staphylococcus aureus

    Directory of Open Access Journals (Sweden)

    Henrique D. M. Coutinho

    2015-09-01

    Full Text Available Introduction: Combining multiple drugs is an strategy used to combat the dissemination of pathogenic and drug resistant bacteria. However, the misuse of these drugs against bacteria have caused the selection of more resistant specimens called multidrug-resistant bacteria. Objective: In this work we evaluated the antibiotic activity of claritromicin, gentamicin, ciprofloxacin and imipenen, alone or associating one by one, against strains of Escherichia coli and Staphylococcus aureus. Material and methods: The minimal inhibitory concentration (mic was performed us­ing the microdilution assay. Based in the mic values, the antibiotic effect of the drugs alone and in association were determined. Results: The association between the drugs demonstrated the synergism against the bacterial strains. Conclusion: The use of the combined antibiotic-therapy can be positively performed, but additional studies have to be conducted first for proving that its use is safe.

  7. Evidence that the phr+ gene enhances the ultraviolet resistance of Escherichia coli recA strains in the dark.

    Science.gov (United States)

    Yamamoto, K; Fujiwara, Y; Shinagawa, H

    1983-01-01

    An Escherichia coli recA phr+ purA strain was more resistant to ultraviolet radiation than its isogenic derivative recA phr+ purA+ in the absence of photoreactivating light, whereas their nearly isogenic derivative recA phr showed most UV-induced lethality. The amounts of photoreactivating enzyme (PRE) per cell in the recA phr+ purA was higher than in the recA phr+ purA+. The recA phr is defective for photoreactivation. Thus, in the recA strain, UV resistance in the dark increased in proportion to the amounts of PRE per cell, suggesting that PRE participates in the process of dark repair of UV-damaged DNA.

  8. Detection by molecular hybridization of pap, afa, and sfa adherence systems in Escherichia coli strains associated with urinary and enteral infections.

    Science.gov (United States)

    Archambaud, M; Courcoux, P; Labigne-Roussel, A

    1988-01-01

    The genetic determinants responsible for the adherence of Escherichia coli to uroepithelial cells have been identified in recent years by genetic and molecular methods. Specific DNA probes for each of the three operons which have been cloned so far (pap, afa, sfa/foc operons) have been used in colony hybridization experiments to detect the presence of each of these operons in the chromosomal DNA of 443 strains of E. coli; 186 strains were from patients with urinary tract infections (pyelonephritis, 106 strains; cystitis, 59; asymptomatic bacteriuria, 21) and 257 were strains from the stools of healthy subjects (61) or from patients with various enteral infections (196). E. coli strains harbouring the pap operon were found more frequently in the urine of patients with pyelonephritis (p less than 0.001) and cystitis (p less than 0.01) than in control stools. The presence of two operons (pap + afa) or (pap + sfa/foc) was only observed in uropathogenic strains (p less than 0.02). Pap and sfa/foc operons were never found in strains causing enteral infection; however, the afa operon was found in 7.6% of the enteropathogenic E. coli.

  9. Adherence to abiotic surface induces SOS response in Escherichia coli K-12 strains under aerobic and anaerobic conditions.

    Science.gov (United States)

    Costa, Suelen B; Campos, Ana Carolina C; Pereira, Ana Claudia M; de Mattos-Guaraldi, Ana Luiza; Júnior, Raphael Hirata; Rosa, Ana Cláudia P; Asad, Lídia M B O

    2014-09-01

    During the colonization of surfaces, Escherichia coli bacteria often encounter DNA-damaging agents and these agents can induce several defence mechanisms. Base excision repair (BER) is dedicated to the repair of oxidative DNA damage caused by reactive oxygen species (ROS) generated by chemical and physical agents or by metabolism. In this work, we have evaluated whether the interaction with an abiotic surface by mutants derived from E. coli K-12 deficient in some enzymes that are part of BER causes DNA damage and associated filamentation. Moreover, we studied the role of endonuclease V (nfi gene; 1506 mutant strain) in biofilm formation. Endonuclease V is an enzyme that is involved in DNA repair of nitrosative lesions. We verified that endonuclease V is involved in biofilm formation. Our results showed more filamentation in the xthA mutant (BW9091) and triple xthA nfo nth mutant (BW535) than in the wild-type strain (AB1157). By contrast, the mutant nfi did not present filamentation in biofilm, although its wild-type strain (1466) showed rare filaments in biofilm. The filamentation of bacterial cells attaching to a surface was a consequence of SOS induction measured by the SOS chromotest. However, biofilm formation depended on the ability of the bacteria to induce the SOS response since the mutant lexA Ind(-) did not induce the SOS response and did not form any biofilm. Oxygen tension was an important factor for the interaction of the BER mutants, since these mutants exhibited decreased quantitative adherence under anaerobic conditions. However, our results showed that the presence or absence of oxygen did not affect the viability of BW9091 and BW535 strains. The nfi mutant and its wild-type did not exhibit decreased biofilm formation under anaerobic conditions. Scanning electron microscopy was also performed on the E. coli K-12 strains that had adhered to the glass, and we observed the presence of a structure similar to an extracellular matrix that depended on the

  10. Characterization of ESBL-producing Escherichia coli and Klebsiella pneumoniae strains isolated from urine of nonhospitalized patients in the Zagreb region

    Directory of Open Access Journals (Sweden)

    Branka Bedenić,

    2010-02-01

    Full Text Available Aim To determine the prevalence of ESBL-producing Escherichia coli and Klebsiella pneumoniae strains isolated from urine of nonhospitalized patients during a three-year period, to determine their antibiotic susceptibility, investigate the transfer of ESBL genes with cotransfer of resistance and to characterize isolated beta-lactamases. Methods Antimicrobial susceptibility was determined by disk diffusion and broth microdilution methods. The double-disk test was used for ESBL detection. Transfer of resistance was performed by broth mating method and characterization of isolated beta-lactamases by polymerase chain reaction. Results The prevalence of ESBL-producing E. coli was 1.5% and of K. pneumoniae 4.1% with its different distribution according to patients`age and gender. ESBL-producing K. pneumoniae showed high resistance rates to aminoglycosides, cotrimoxazole, nitrofurantoin and quinolones while ESBL-producing E. coli isolates, with exception of high aminoglycoside resistance, showed low resistance rates to other antibiotics. Successful conjugation of ESBL genes was obtained with 25% E. coli and 76.2% K. pneumoniae strains. Comparing to E. coli, K. pneumoniae strains showed higher rates of aminoglycosideand cotrimoxazole resistance cotransfer. Beta-lactamases of investigated strains belonged to TEM, SHV and CTX-M families.Conclusion The existence of multiple-resistant ESBL-producing E. coli and K. pneumoniae strains was confirmed in observed outpatient population. ESBL-producing K. pneumoniae isolates, in contrast toESBL-producing E. coli, showed higher resistance rates to non-beta-lactam antibiotics, probably caused by cotransfer of resistance genes located on the same plasmid as ESBL genes. It is important to monitor the prevalence of such strains and their possible spreading in the outpatient population of the Zagreb region

  11. High Incidence of Escherichia coli Strains Coharboring mcr-1 and blaNDM from Chickens.

    Science.gov (United States)

    Liu, Bao-Tao; Song, Feng-Jing; Zou, Ming; Zhang, Qi-Di; Shan, Hu

    2017-03-01

    This study investigated the characteristics of Escherichia coli isolates carrying mcr-1-blaNDM from a chicken farm in China. Of the 78 E. coli isolates, 21 clonally unrelated isolates carried mcr-1-blaNDM Diverse IncI2 plasmids disseminated mcr-1, while the dissemination of blaNDM was mediated by diverse IncB/O plasmids. More striking was the colocalization of resistance genes mcr-1 and blaNDM-4 in an IncHI2/ST3 plasmid, which might pose a great challenge for public health. Copyright © 2017 American Society for Microbiology.

  12. Shiga toxin-producing Escherichia coli strains isolated from dairy products - Genetic diversity and virulence gene profiles.

    Science.gov (United States)

    Douëllou, T; Delannoy, S; Ganet, S; Mariani-Kurkdjian, P; Fach, P; Loukiadis, E; Montel, Mc; Thevenot-Sergentet, D

    2016-09-02

    Shiga toxin-producing Escherichia coli (STEC) are widely recognized as pathogens causing food borne disease. Here we evaluate the genetic diversity of 197 strains, mainly STEC, from serotypes O157:H7, O26:H11, O103:H2, O111:H8 and O145:28 and compared strains recovered in dairy products against strains from human, meat and environment cases. For this purpose, we characterized a set of reference-collection STEC isolates from dairy products by PFGE DNA fingerprinting and a subset of these by virulence-gene profiling. PFGE profiles of restricted STEC total DNA showed high genomic variability (0.9976 on Simpson's discriminatory index), enabling all dairy isolates to be differentiated. High-throughput real-time PCR screening of STEC virulence genes were applied on the O157:H7 and O26:H11 STEC isolates from dairy products and human cases. The virulence gene profiles of dairy and human STEC strains were similar. Nevertheless, frequency-wise, stx1 was more prevalent among dairy O26:H11 isolates than in human cases ones (87% vs. 44%) while stx2 was more prevalent among O26:H11 human isolates (23% vs. 81%). For O157:H7 isolates, stx1 (0% vs. 39%), nleF (40% vs 94%) and Z6065 (40% vs 100%) were more prevalent among human than dairy strains. Our data point to differences between human and dairy strains but these differences were not sufficient to associate PFGE and virulence gene profiles to a putative lower pathogenicity of dairy strains based on their lower incidence in disease. Further comparison of whole-genome expression and virulence gene profiles should be investigated in cheese and intestinal tract samples.

  13. Mechanisms of tolerance and high degradation capacity of the herbicide mesotrione by Escherichia coli strain DH5-α.

    Directory of Open Access Journals (Sweden)

    Luiz R Olchanheski

    Full Text Available The intensive use of agrochemicals has played an important role in increasing agricultural production. One of the impacts of agrochemical use has been changes in population structure of soil microbiota. The aim of this work was to analyze the adaptive strategies that bacteria use to overcome oxidative stress caused by mesotrione, which inhibits 4-hydroxyphenylpyruvate dioxygenase. We also examined antioxidative stress systems, saturation changes of lipid membranes, and the capacity of bacteria to degrade mesotrione. Escherichia coli DH5-á was chosen as a non-environmental strain, which is already a model bacterium for studying metabolism and adaptation. The results showed that this bacterium was able to tolerate high doses of the herbicide (10× field rate, and completely degraded mesotrione after 3 h of exposure, as determined by a High Performance Liquid Chromatography. Growth rates in the presence of mesotrione were lower than in the control, prior to the period of degradation, showing toxic effects of this herbicide on bacterial cells. Changes in the saturation of the membrane lipids reduced the damage caused by reactive oxygen species and possibly hindered the entry of xenobiotics in the cell, while activating glutathione-S-transferase enzyme in the antioxidant system and in the metabolizing process of the herbicide. Considering that E. coli DH5-α is a non-environmental strain and it had no previous contact with mesotrione, the defense system found in this strain could be considered non-specific. This bacterium system response may be a general adaptation mechanism by which bacterial strains resist to damage from the presence of herbicides in agricultural soils.

  14. Molecular typing of Escherichia coli strains associated with threatened sea ducks and near-shore marine habitats of south-west Alaska

    Science.gov (United States)

    Hollmén, Tuula E.; Debroy, C.; Flint, P.L.; Safine, D.E.; Schamber, J.L.; Riddle, A.E.; Trust, K.A.

    2011-01-01

    In Alaska, sea ducks winter in coastal habitats at remote, non-industrialized areas, as well as in proximity to human communities and industrial activity. We evaluated prevalence and characteristics of Escherichia coli strains in faecal samples of Steller's eiders (Polysticta stelleri; n=122) and harlequin ducks (Histrionicus histrionicus; n=21) at an industrialized site and Steller's eiders (n=48) at a reference site, and compared these strains with those isolated from water samples from near-shore habitats of ducks. The overall prevalence of E. coli was 16% and 67% in Steller's eiders and harlequin ducks, respectively, at the industrialized study site, and 2% in Steller's eiders at the reference site. Based on O and H antigen subtyping and genetic characterization by enterobacterial repetitive intergenic consensus polymerase chain reaction and pulsed-field gel electrophoresis, we found evidence of avian pathogenic E. coli (APEC) strains associated with both species and detected E. coli strains carrying virulence genes associated with mammals in harlequin ducks. Steller's eiders that carried APEC had lower serum total protein and albumin concentrations, providing further evidence of pathogenicity. The genetic profile of two E. coli strains from water matched an isolate from a Steller's eider providing evidence of transmission between near-shore habitats and birds. ?? 2010 Society for Applied Microbiology and Blackwell Publishing Ltd.

  15. Developmental pathway for biofilm formation in curli-producing Escherichia coli strains: role of flagella, curli and colanic acid.

    Science.gov (United States)

    Prigent-Combaret, C; Prensier, G; Le Thi, T T; Vidal, O; Lejeune, P; Dorel, C

    2000-08-01

    This work was performed to establish a model describing bacterial surface structures involved in biofilm development, in curli-overproducing Escherichia coli K-12 strains, at 30 degrees C, and in minimal growth medium. Using a genetic approach, in association with observations of sessile communities by light and electron microscopic techniques, the role of protein surface structures, such as flagella and curli, and saccharidic surface components, such as the E. coli exopolysaccharide, colanic acid, was determined. We show that, in the context of adherent ompR234 strains, (i) flagellar motility is not required for initial adhesion and biofilm development; (ii) both primary adhesion to inert surfaces and development of multilayered cell clusters require curli synthesis; (iii) curli display direct interactions with the substratum and form interbacterial bundles, allowing a cohesive and stable association of cells; and (iv) colanic acid does not appear critical for bacterial adhesion and further biofilm development but contributes to the biofilm architecture and allows for the formation of voluminous biofilms.

  16. Changes in the proteome of Mastitis-causing escherichia coli strains that affect pathogenesis

    Science.gov (United States)

    Escherichia coli is a leading cause of bacterial mastitis in dairy cattle. Milk is the environment in which bacteria must grow to establish an infection of the mammary gland. However, milk is not a rich growth media for bacteria. In fact, milk naturally contains many mechanisms to inhibit bacterial ...

  17. [A selective nutrient medium for isolating clinical strains of Escherichia coli O157:H7].

    Science.gov (United States)

    Sultanov, Z Z; Stepanova, E D; Kakulina, E A

    2000-01-01

    A dried selective culture medium, electrolyte-deficient sorbitol agar (EDS agar), for the isolation and preliminary identification of E. coli O157:H7 from clinical material has been developed. The medium is not inferior in its quality to analogous foreign media and requires no scarce ingredients for its manufacture.

  18. Characterization of toxin from Verocytotoxigenic Ecscherichia coli (VTEC)strains isolated from neonatal calves in India

    Institute of Scientific and Technical Information of China (English)

    Diganta Pan; Ashok Kumar Bhatia; Bhilegaonkar KN

    2009-01-01

    Objective:The present study has characterized dialyzed toxin from non-O157 VTEC E.coli isolates by vero cell toxicity assay and pathogenecity in mice model.Methods:Toxins from non-O157 verocytotoxic Escherichi-a coli isolated from neonatal calves were characterized.Dialyzed toxin from E.coli O26,O111 and O103 sero-types were prepared and characterized by verocell toxicity assay and pathogenicity in mice model.E.coli O157:H7 considered as positive control for this study.Results:Cytopathic effects in vero cell line first roun-ding of vero cells,followed by clumping of cells and finally disintegrated,blackened,shriveled cell line within 16 to 72 hrs.Phenotypic markers such as hind limb paralysis and reddening of tail were prominent in all the toxicated mice.Extensive histopathological study was conducted for multiple organ involvement.Conclusion:Several methods for toxin assay were developed based on biological,immunological and detection of virulence genes related to toxin production but each test has draw back.Therefore,it is likely that future effort will be focused on the development of assay,which is fast,reliable,specific and sensitive methods based on mice model.

  19. Pulsed-field gel electrophoresis typing of Escherichia coli strains from samples collected before and after pivmecillinam or placebo treatment of uncomplicated community-acquired urinary tract infection in women

    DEFF Research Database (Denmark)

    Ejrnaes, Karen; Sandvang, Dorthe; Lundgren, Bettina

    2006-01-01

    The primary infecting Escherichia coli strains from 156 women with community-acquired uncomplicated urinary tract infection (UTI) randomized to pivmecillinam or placebo and the E. coli strains causing UTI at two follow-up visits were typed using pulsed-field gel electrophoresis (PFGE). In the piv......The primary infecting Escherichia coli strains from 156 women with community-acquired uncomplicated urinary tract infection (UTI) randomized to pivmecillinam or placebo and the E. coli strains causing UTI at two follow-up visits were typed using pulsed-field gel electrophoresis (PFGE...

  20. Pathogenic potential, genetic diversity, and population structure of Escherichia coli strains isolated from a forest-dominated watershed (Comox Lake) in British Columbia, Canada.

    Science.gov (United States)

    Chandran, Abhirosh; Mazumder, Asit

    2015-03-01

    Escherichia coli isolates (n = 658) obtained from drinking water intakes of Comox Lake (2011 to 2013) were screened for the following virulence genes (VGs): stx1 and stx2 (Shiga toxin-producing E. coli [STEC]), eae and the adherence factor (EAF) gene (enteropathogenic E. coli [EPEC]), heat-stable (ST) enterotoxin (variants STh and STp) and heat-labile enterotoxin (LT) genes (enterotoxigenic E. coli [ETEC]), and ipaH (enteroinvasive E. coli [EIEC]). The only genes detected were eae and stx2, which were carried by 37.69% (n = 248) of the isolates. Only eae was harbored by 26.74% (n = 176) of the isolates, representing potential atypical EPEC strains, while only stx2 was detected in 10.33% (n = 68) of the isolates, indicating potential STEC strains. Moreover, four isolates were positive for both the stx2 and eae genes, representing potential EHEC strains. The prevalence of VGs (eae or stx2) was significantly (P < 0.0001) higher in the fall season, and multiple genes (eae plus stx2) were detected only in fall. Repetitive element palindromic PCR (rep-PCR) fingerprint analysis of 658 E. coli isolates identified 335 unique fingerprints, with an overall Shannon diversity (H') index of 3.653. Diversity varied among seasons over the years, with relatively higher diversity during fall. Multivariate analysis of variance (MANOVA) revealed that the majority of the fingerprints showed a tendency to cluster according to year, season, and month. Taken together, the results indicated that the diversity and population structure of E. coli fluctuate on a temporal scale, reflecting the presence of diverse host sources and their behavior over time in the watershed. Furthermore, the occurrence of potentially pathogenic E. coli strains in the drinking water intakes highlights the risk to human health associated with direct and indirect consumption of untreated surface water.

  1. An epidemiological study on the drug resistance of Escherichia coli strains isolated from women patients with urinary tract infection in Shalamzar, Iran

    Directory of Open Access Journals (Sweden)

    Gholamreza Farnoosh

    2015-03-01

    Full Text Available Objective: To investigate drug resistance of various strains of Escherichia coli (E. coli bacteria isolated from female patients with urinary tracts infections (UTIs in Shalamzar, Iran. Methods: This study was conducted from April 2011 to April 2012 on 150 female patients with positive urine culture and 105 CFU/mL colony count. The pattern of antibiotic sensitivity was recognized using antibiogram by the disc diffusion method. Results: The results revealed that the predominant bacterium was E. coli (90%, followed by Klebsiella pneumonia (3%. Trimethoprim-sulfamethoxazole is the initial medicine to treat UTIs (without complications which demonstrated relatively poor activity against E. coli (with 40% sensitivity, though alternative medicines such as nitrofurantoin (97% sensitivity and ciprofloxacin (91% sensitivity showed good activity against E. coli as well. Conclusions: The findings emphasized the necessity of pursuing the investigations in national and local governments in order to retain the efficacy of treating UTIs using effective antibiotics.

  2. Mutant prevention concentrations of pradofloxacin for susceptible and mutant strains of Escherichia coli with reduced fluoroquinolone susceptibility.

    Science.gov (United States)

    Marcusson, Linda L; Komp Lindgren, Patricia; Olofsson, Sara K; Hughes, Diarmaid; Cars, Otto

    2014-10-01

    Pharmacodynamic and mutant prevention properties of the fluoroquinolone pradofloxacin (PRA) were measured against a set of 17 Escherichia coli strains carrying no, one or two known mutations conferring reduced fluoroquinolone susceptibility. The strains included susceptible wild-types, isogenic constructed mutants, isogenic selected mutants and clinical isolates. The effectiveness of PRA was determined with regard to preventing the selection of resistant mutants, using static and changing concentrations of drug. Ciprofloxacin was used as a reference drug. Minimum inhibitory concentrations (MICs) and mutant prevention concentrations (MPCs) of PRA for the susceptible wild-type strains were in the range 0.012-0.016mg/L and 0.2-0.3mg/L, respectively, giving a mean±standard deviation mutant prevention index (MPI=MPC/MIC) of 17.7±1.1. The mean MPI PRA of the 14 mutant strains was 19.2±12, and the mean MPI across all 17 strains was 18.9±10.8. In an in vitro kinetic model in which PRA was diluted with a half-life of 7h to mimic in vivo conditions, an initial concentration of PRA of 1.6-2.4mg/L (8-10× MPC), giving a PRA AUC/MPC ratio of 73-92, and a T>MPC of 21-23h was sufficient to prevent the selection of resistant mutants from the three susceptible wild-type strains. Dosing to reduce selection for antibiotic resistance in veterinary therapy has a role in reducing the reservoir of resistant mutants. We conclude that a level of dosing that prevents the selection of resistant mutants during therapy should be achievable in vivo. Copyright © 2014 Elsevier B.V. and the International Society of Chemotherapy. All rights reserved.

  3. Lineages and virulence gene content among extended-spectrum β-lactamase-producing Escherichia coli strains of food origin in Tunisia.

    Science.gov (United States)

    Jouini, Ahlem; Slama, Karim Ben; Klibi, Naouel; Sallem, Rym Ben; Estepa, Vanesa; Vinué, Laura; Sáenz, Yolanda; Ruiz-Larrea, Fernanda; Boudabous, Abdellatif; Torres, Carmen

    2013-02-01

    Nineteen extended-spectrum β-lactamase (ESBL)-positive Escherichia coli strains recovered from food samples in Tunisia were characterized by multilocus sequence typing and phylogenetic typing, and the virulence gene and plasmid content were also determined. These strains presented unrelated pulsed-field gel electrophoresis patterns and contained genes coding for the following ESBLs (the number of strains is in parentheses): CTX-M-1 (15), CTX-M-14 (2), CTX-M-8 (1), and SHV-5 (1). Twelve different sequence types (STs) were identified among the 19 ESBL-positive strains, which included two new STs (ST2022 in 2 bla(CTX-M-14)-containing strains and ST1970 in 2 bla(CTX-M-1)-containing strains). ST155 and ST602 were detected in four and three bla(CTX-M-1)-containing strains, respectively, and ST405 was detected in one bla(CTX-M-8)-producing strain. All ESBL-positive strains were ascribed to the phylogenetic groups A and B1. Most of the bla(CTX-M-1)-containing strains harbored an IncI1 plasmid, except for the four bla(CTX-M-1)-positive strains of beef origin and ST155, which harbored an IncN plasmid. The two bla(CTX-M-14)-containing strains contained an IncI1 plasmid. The virulence gene fimA was detected in all strains. Most strains also carried the aer gene, and six strains carried the eae gene. All strains were negative for the virulence genes sxt, papG-III, papC, hly, cnf1, and bfp. We conclude that ESBL-producing E. coli strains of food origin in Tunisia show high diversity and that plasmids harboring ESBL genes could be implicated in the dissemination of this resistance phenotype.

  4. Identification of Extended-Spectrum β-Lactamases Escherichia coli Strains Isolated from Market Garden Products and Irrigation Water in Benin.

    Science.gov (United States)

    Moussé, Wassiyath; Sina, Haziz; Baba-Moussa, Farid; Noumavo, Pacôme A; Agbodjato, Nadège A; Adjanohoun, Adolphe; Baba-Moussa, Lamine

    2015-01-01

    The present study aimed at biochemical and molecular characterization of Escherichia coli strains isolated from horticultural products and irrigation water of Cotonou. The samples were collected from 12 market gardeners of 4 different sites. Rapid' E. coli medium was used for identification of E. coli strains and the antimicrobial susceptibility was performed by the agar disk diffusion method. The β-lactamases production was sought by the liquid acidimetric method. The genes coding for β-lactamases and toxins were identified by PCR method. The results revealed that about 34.95% of the analyzed samples were contaminated by E. coli. Cabbages were the most contaminated by E. coli (28.26%) in dry season. All isolated strains were resistant to amoxicillin. The penicillinase producing E. coli carried blaTEM (67.50%), blaSHV (10%), and blaCTX-M (22.50%) genes. The study revealed that the resistance genes such as SLTI (35.71%), SLTII (35.71%), ETEC (7.15%), and VTEC (21.43%) were carried. Openly to the found results and considering the importance of horticultural products in Beninese food habits, it is important to put several strategies aiming at a sanitary security by surveillance and sensitization of all the actors on the risks of some practices.

  5. Metabolic engineering for the production of shikimic acid in an evolved Escherichia coli strain lacking the phosphoenolpyruvate: carbohydrate phosphotransferase system

    Directory of Open Access Journals (Sweden)

    Bolívar Francisco

    2010-04-01

    Full Text Available Abstract Background Shikimic acid (SA is utilized in the synthesis of oseltamivir-phosphate, an anti-influenza drug. In this work, metabolic engineering approaches were employed to produce SA in Escherichia coli strains derived from an evolved strain (PB12 lacking the phosphoenolpyruvate:carbohydrate phosphotransferase system (PTS- but with capacity to grow on glucose. Derivatives of PB12 strain were constructed to determine the effects of inactivating aroK, aroL, pykF or pykA and the expression of plasmid-coded genes aroGfbr, tktA, aroB and aroE, on SA synthesis. Results Batch cultures were performed to evaluate the effects of genetic modifications on growth, glucose consumption, and aromatic intermediate production. All derivatives showed a two-phase growth behavior with initial high specific growth rate (μ and specific glucose consumption rate (qs, but low level production of aromatic intermediates. During the second growth phase the μ decreased, whereas aromatic intermediate production reached its maximum. The double aroK- aroL- mutant expressing plasmid-coded genes (strain PB12.SA22 accumulated SA up to 7 g/L with a yield of SA on glucose of 0.29 mol/mol and a total aromatic compound yield (TACY of 0.38 mol/mol. Single inactivation of pykF or pykA was performed in PB12.SA22 strain. Inactivation of pykF caused a decrease in μ, qs, SA production, and yield; whereas TACY increased by 33% (0.5 mol/mol. Conclusions The effect of increased availability of carbon metabolites, their channeling into the synthesis of aromatic intermediates, and disruption of the SA pathway on SA production was studied. Inactivation of both aroK and aroL, and transformation with plasmid-coded genes resulted in the accumulation of SA up to 7 g/L with a yield on glucose of 0.29 mol/mol PB12.SA22, which represents the highest reported yield. The pykF and pykA genes were inactivated in strain PB12.SA22 to increase the production of aromatic compounds in the PTS

  6. Probiotic Escherichia coli strain Nissle 1917 outcompetes intestinal pathogens during biofilm formation

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Dahl, M.; Klemm, Per

    2010-01-01

    Nissle 1917 has been used for many decades as a probiotic against a variety of intestinal disorders and is probably the best field-tested E. coil strain in the world. Here we have investigated the biofilm-forming capacity of Nissle 1917. We found that the strain was a good biofilm former. Not only...

  7. Suppression of initiation-negative strains of Escherichia coli by integration of the sex factor F.

    Science.gov (United States)

    Tresguerres, E F; Nandadasa, H G; Pritchard, R H

    1975-01-01

    Data are presented suggesting that the most critical factor determining whether an Hfr dnaAts strain can synthesize deoxyribonucleic acid and form colonies at temperatures that are nonpermissive for corresponding F- strains is neither the site of insertion of F nor the presence of additional mutations in the F particle or the chromosome; it is whether the particle is capable of autonomous replication at the temperature used. Consequently, suppression of the DnaA phenotype in Hfr strains occurs at 40 C but not, in most of them, at 42 C without the occurrence of additional mutations. The site of insertion of F may also be important since it is shown that in one Hfr dnaA strain partial suppression does occur at 42 C. In addition, it is shown that strains exhibiting suppression by integration of F at 40 C on minimal agar plates do not do so at this temperature on nutrient agar plates. PMID:1089635

  8. The Adherent/Invasive Escherichia coli Strain LF82 Invades and Persists in Human Prostate Cell Line RWPE-1, Activating a Strong Inflammatory Response

    Science.gov (United States)

    Aleandri, Marta; Marazzato, Massimiliano; Conte, Antonietta L.; Ambrosi, Cecilia; Nicoletti, Mauro; Zagaglia, Carlo; Gambara, Guido; Palombi, Fioretta; De Cesaris, Paola; Ziparo, Elio; Palamara, Anna T.; Riccioli, Anna

    2016-01-01

    Adherent/invasive Escherichia coli (AIEC) strains have recently been receiving increased attention because they are more prevalent and persistent in the intestine of Crohn's disease (CD) patients than in healthy subjects. Since AIEC strains show a high percentage of similarity to extraintestinal pathogenic E. coli (ExPEC), neonatal meningitis-associated E. coli (NMEC), and uropathogenic E. coli (UPEC) strains, here we compared AIEC strain LF82 with a UPEC isolate (strain EC73) to assess whether LF82 would be able to infect prostate cells as an extraintestinal target. The virulence phenotypes of both strains were determined by using the RWPE-1 prostate cell line. The results obtained indicated that LF82 and EC73 are able to adhere to, invade, and survive within prostate epithelial cells. Invasion was confirmed by immunofluorescence and electron microscopy. Moreover, cytochalasin D and colchicine strongly inhibited bacterial uptake of both strains, indicating the involvement of actin microfilaments and microtubules in host cell invasion. Moreover, both strains belong to phylogenetic group B2 and are strong biofilm producers. In silico analysis reveals that LF82 shares with UPEC strains several virulence factors: namely, type 1 pili, the group II capsule, the vacuolating autotransporter toxin, four iron uptake systems, and the pathogenic island (PAI). Furthermore, compared to EC73, LF82 induces in RWPE-1 cells a marked increase of phosphorylation of mitogen-activated protein kinases (MAPKs) and of NF-κB already by 5 min postinfection, thus inducing a strong inflammatory response. Our in vitro data support the hypothesis that AIEC strains might play a role in prostatitis, and, by exploiting host-cell signaling pathways controlling the innate immune response, likely facilitate bacterial multiplication and dissemination within the male genitourinary tract. PMID:27600504

  9. Comparative genomic analysis of two novel sporadic Shiga toxin-producing Escherichia coli O104:H4 strains isolated 2011 in Germany.

    Directory of Open Access Journals (Sweden)

    Erhard Tietze

    Full Text Available A large outbreak of gastrointestinal disease occurred in 2011 in Germany which resulted in almost 4000 patients with acute gastroenteritis or hemorrhagic colitis, 855 cases of a hemolytic uremic syndrome and 53 deaths. The pathogen was an uncommon, multiresistant Escherichia coli strain of serotype O104:H4 which expressed a Shiga toxin characteristic of enterohemorrhagic E. coli and in addition virulence factors common to enteroaggregative E. coli. During post-epidemic surveillance of Shiga toxin-producing E. coli (STEC all but two of O104:H4 isolates were indistinguishable from the epidemic strain. Here we describe two novel STEC O104:H4 strains isolated in close spatiotemporal proximity to the outbreak which show a virulence gene panel, a Shiga toxin-mediated cytotoxicity towards Vero cells and aggregative adherence to Hep-2 cells comparable to the outbreak strain. They differ however both from the epidemic strain and from each other, by their antibiotic resistance phenotypes and some other features as determined by routine epidemiological subtyping methods. Whole genome sequencing of these two strains, of ten outbreak strain isolates originating from different time points of the outbreak and of one historical sporadic EHEC O104:H4 isolate was performed. Sequence analysis revealed a clear phylogenetic distance between the two variant strains and the outbreak strain finally identifying them as epidemiologically unrelated isolates from sporadic cases. These findings add to the knowledge about this emerging pathogen, illustrating a certain diversity within the bacterial core genome as well as loss and gain of accessory elements. Our results do also support the view that distinct new variants of STEC O104:H4 repeatedly might originate from yet unknown reservoirs, rather than that there would be a continuous diversification of a single epidemic strain established and circulating in Germany after the large outbreak in 2011.

  10. Comparative genomic analysis of two novel sporadic Shiga toxin-producing Escherichia coli O104:H4 strains isolated 2011 in Germany.

    Science.gov (United States)

    Tietze, Erhard; Dabrowski, Piotr Wojciech; Prager, Rita; Radonic, Aleksandar; Fruth, Angelika; Auraß, Philipp; Nitsche, Andreas; Mielke, Martin; Flieger, Antje

    2015-01-01

    A large outbreak of gastrointestinal disease occurred in 2011 in Germany which resulted in almost 4000 patients with acute gastroenteritis or hemorrhagic colitis, 855 cases of a hemolytic uremic syndrome and 53 deaths. The pathogen was an uncommon, multiresistant Escherichia coli strain of serotype O104:H4 which expressed a Shiga toxin characteristic of enterohemorrhagic E. coli and in addition virulence factors common to enteroaggregative E. coli. During post-epidemic surveillance of Shiga toxin-producing E. coli (STEC) all but two of O104:H4 isolates were indistinguishable from the epidemic strain. Here we describe two novel STEC O104:H4 strains isolated in close spatiotemporal proximity to the outbreak which show a virulence gene panel, a Shiga toxin-mediated cytotoxicity towards Vero cells and aggregative adherence to Hep-2 cells comparable to the outbreak strain. They differ however both from the epidemic strain and from each other, by their antibiotic resistance phenotypes and some other features as determined by routine epidemiological subtyping methods. Whole genome sequencing of these two strains, of ten outbreak strain isolates originating from different time points of the outbreak and of one historical sporadic EHEC O104:H4 isolate was performed. Sequence analysis revealed a clear phylogenetic distance between the two variant strains and the outbreak strain finally identifying them as epidemiologically unrelated isolates from sporadic cases. These findings add to the knowledge about this emerging pathogen, illustrating a certain diversity within the bacterial core genome as well as loss and gain of accessory elements. Our results do also support the view that distinct new variants of STEC O104:H4 repeatedly might originate from yet unknown reservoirs, rather than that there would be a continuous diversification of a single epidemic strain established and circulating in Germany after the large outbreak in 2011.

  11. The constancy of global regulation across a species: the concentrations of ppGpp and RpoS are strain-specific in Escherichia coli

    Directory of Open Access Journals (Sweden)

    Phan Katherine

    2011-03-01

    Full Text Available Abstract Background Sigma factors and the alarmone ppGpp control the allocation of RNA polymerase to promoters under stressful conditions. Both ppGpp and the sigma factor σS (RpoS are potentially subject to variability across the species Escherichia coli. To find out the extent of strain variation we measured the level of RpoS and ppGpp using 31 E. coli strains from the ECOR collection and one reference K-12 strain. Results Nine ECORs had highly deleterious mutations in rpoS, 12 had RpoS protein up to 7-fold above that of the reference strain MG1655 and the remainder had comparable or lower levels. Strain variation was also evident in ppGpp accumulation under carbon starvation and spoT mutations were present in several low-ppGpp strains. Three relationships between RpoS and ppGpp levels were found: isolates with zero RpoS but various ppGpp levels, strains where RpoS levels were proportional to ppGpp and a third unexpected class in which RpoS was present but not proportional to ppGpp concentration. High-RpoS and high-ppGpp strains accumulated rpoS mutations under nutrient limitation, providing a source of polymorphisms. Conclusions The ppGpp and σS variance means that the expression of genes involved in translation, stress and other traits affected by ppGpp and/or RpoS are likely to be strain-specific and suggest that influential components of regulatory networks are frequently reset by microevolution. Different strains of E. coli have different relationships between ppGpp and RpoS levels and only some exhibit a proportionality between increasing ppGpp and RpoS levels as demonstrated for E. coli K-12.

  12. Functional genomics of probiotic Escherichia coli Nissle 1917 and 83972, and UPEC strain CFT073: comparison of transcriptomes, growth and biofilm formation

    DEFF Research Database (Denmark)

    Hancock, Viktoria; Vejborg, Rebecca Munk; Klemm, Per

    2010-01-01

    Strain CFT073 is a bona fide uropathogen, whereas strains 83972 and Nissle 1917 are harmless probiotic strains of urinary tract and faecal origin, respectively. Despite their different environmental origins and dispositions the three strains are very closely related and the ancestors of 83972...... of the three strains are closely related. The data demonstrate that the distance from a pathogen to a probiotic strain can be surprisingly short. We demonstrate that Nissle 1917, in spite of its intestinal niche origin, grows well in urine, and is a good biofilm former in this medium in which it also out...... accounting for the impaired biofilm formation. Although the three strains have very different strategies vis-à-vis the human host their functional gene profiles are surprisingly similar. It is also interesting to note that the only two Escherichia coli strains used as probiotics are in fact deconstructed...

  13. Genetic Diversity and Virulence Potential of Shiga Toxin-Producing Escherichia coli O113:H21 Strains Isolated from Clinical, Environmental, and Food Sources

    Science.gov (United States)

    Delannoy, Sabine; Lacher, David W.; dos Santos, Luis Fernando; Beutin, Lothar; Fach, Patrick; Rivas, Marta; Hartland, Elizabeth L.; Paton, Adrienne W.; Guth, Beatriz E. C.

    2014-01-01

    Shiga toxin-producing Escherichia coli strains of serotype O113:H21 have caused severe human diseases, but they are unusual in that they do not produce adherence factors coded by the locus of enterocyte effacement. Here, a PCR microarray was used to characterize 65 O113:H21 strains isolated from the environment, food, and clinical infections from various countries. In comparison to the pathogenic strains that were implicated in hemolytic-uremic syndrome in Australia, there were no clear differences between the pathogens and the environmental strains with respect to the 41 genetic markers tested. Furthermore, all of the strains carried only Shiga toxin subtypes associated with human infections, suggesting that the environmental strains have the potential to cause disease. Most of the O113:H21 strains were closely related and belonged in the same clonal group (ST-223), but CRISPR analysis showed a great degree of genetic diversity among the O113:H21 strains. PMID:24858089

  14. Complete genome sequence and comparative analysis of the wild-type commensal Escherichia coli strain SE11 isolated from a healthy adult.

    Science.gov (United States)

    Oshima, Kenshiro; Toh, Hidehiro; Ogura, Yoshitoshi; Sasamoto, Hiroyuki; Morita, Hidetoshi; Park, Sang-Hee; Ooka, Tadasuke; Iyoda, Sunao; Taylor, Todd D; Hayashi, Tetsuya; Itoh, Kikuji; Hattori, Masahira

    2008-12-01

    We sequenced and analyzed the genome of a commensal Escherichia coli (E. coli) strain SE11 (O152:H28) recently isolated from feces of a healthy adult and classified into E. coli phylogenetic group B1. SE11 harbored a 4.8 Mb chromosome encoding 4679 protein-coding genes and six plasmids encoding 323 protein-coding genes. None of the SE11 genes had sequence similarity to known genes encoding phage- and plasmid-borne virulence factors found in pathogenic E. coli strains. The comparative genome analysis with the laboratory strain K-12 MG1655 identified 62 poorly conserved genes between these two non-pathogenic strains and 1186 genes absent in MG1655. These genes in SE11 were mostly encoded in large insertion regions on the chromosome or in the plasmids, and were notably abundant in genes of fimbriae and autotransporters, which are cell surface appendages that largely contribute to the adherence ability of bacteria to host cells and bacterial conjugation. These data suggest that SE11 may have evolved to acquire and accumulate the functions advantageous for stable colonization of intestinal cells, and that the adhesion-associated functions are important for the commensality of E. coli in human gut habitat.

  15. SslE elicits functional antibodies that impair in vitro mucinase activity and in vivo colonization by both intestinal and extraintestinal Escherichia coli strains.

    Directory of Open Access Journals (Sweden)

    Barbara Nesta

    2014-05-01

    Full Text Available SslE, the Secreted and surface-associated lipoprotein from Escherichia coli, has recently been associated to the M60-like extracellular zinc-metalloprotease sub-family which is implicated in glycan recognition and processing. SslE can be divided into two main variants and we recently proposed it as a potential vaccine candidate. By applying a number of in vitro bioassays and comparing wild type, knockout mutant and complemented strains, we have now demonstrated that SslE specifically contributes to degradation of mucin substrates, typically present in the intestine and bladder. Mutation of the zinc metallopeptidase motif of SslE dramatically impaired E. coli mucinase activity, confirming the specificity of the phenotype observed. Moreover, antibodies raised against variant I SslE, cloned from strain IHE3034 (SslEIHE3034, are able to inhibit translocation of E. coli strains expressing different variants through a mucin-based matrix, suggesting that SslE induces cross-reactive functional antibodies that affect the metallopeptidase activity. To test this hypothesis, we used well-established animal models and demonstrated that immunization with SslEIHE3034 significantly reduced gut, kidney and spleen colonization by strains producing variant II SslE and belonging to different pathotypes. Taken together, these data strongly support the importance of SslE in E. coli colonization of mucosal surfaces and reinforce the use of this antigen as a component of a broadly protective vaccine against pathogenic E. coli species.

  16. Pili in Microspheres Protect Rabbits From Diarrhoea Induced by E. Coli Strain RDEC-1

    Science.gov (United States)

    1993-01-01

    antibodies secreted in bile. 14. UIJIT TRMS J~*NUjMBER OF PAGES Keywo- Fntenic vaccine; mucosal immunity ; biliary EgA; mucosal attachment: enteroadherent...vaccine; mucosal immunity ; biliary IgA; mucosal attachment; enteroadherent Escherichia coli INTRODUCTION intestinal tract3 or inadequate uptake of the... Mucosal Immunity and Infections at Mucosal Surfaces (Eds 1986, pp. 143-145 Strober. W., Lamm. M.E., McGhee, J.R. and James, S.P.) Oxford 4 Eldridge

  17. Detection of the CS20 Colonization Factor Antigen in Diffuse-Adhering Escherichia coli Strains

    Science.gov (United States)

    2010-01-01

    identified using molecular methods. FEMS lmmunol Med Microbioi&O (2010) 186-189 FEMS Immunology & Medical Microbiology co 2010 Federation of European...of the gene encoding coli surface antigen 20 of FEMS Immunology & Medical Microbiology co 2010 Federation of European Microbiological Societies...20 1 0) 186-189 FEMS Immunology & Medical Microbiology <.e 2010 Federation of European Microbiological Societies Published by Blackwell Publishing Ltd. No claim to original US government works

  18. C. elegans as a virulence model for E. coli strain 042

    OpenAIRE

    Kjærbo, Rasmus E. R.; Godballe, Troels; Hansen, Klaus G.; Petersen, Pernille D.; Tikander, Emil

    2010-01-01

    During the last decade the nematode Caenorhabditis elegans has been used to model the pathogenesis of several bacterial species. The emerging pathogen enteroaggregative Escherichia coli (EAEC) is a considerable cause of both acute and persistent diarrhea worldwide. Travellers to developing countries, immunocompromised people and young children are high-risk groups prone to infection. Virulence models using C. elegans might provide valuable information about the host-pathogen interactions whic...

  19. Complementation of Escherichia coli unc mutant strains by chloroplast and cyanobacterial F1-ATPase subunits.

    Science.gov (United States)

    Lill, H; Burkovski, A; Altendorf, K; Junge, W; Engelbrecht, S

    1993-10-04

    The genes encoding the five subunits of the F1 portion of the ATPases from both spinach chloroplasts and the cyanobacterium Synechocystis sp. PCC 6803 were cloned into expression vectors and expressed in Escherichia coli. The recombinant subunits formed inclusion bodies within the cells. Each particular subunit was expressed in the respective unc mutant, each unable to grow on non-fermentable carbon sources. The following subunits restored growth under conditions of oxidative phosphorylation: alpha (both sources, cyanobacterial subunit more than spinach subunit), beta (cyanobacterial subunit only), delta (both spinach and Synechocystis), and epsilon (both sources), whereas no growth was achieved with the gamma subunits from both sources. Despite a high degree of sequence homology the large subunits alpha and beta of spinach and cyanobacterial F1 were not as effective in the substitution of their E. coli counterparts. On the other hand, the two smallest subunits of the E. coli ATPase could be more effectively replaced by their cyanobacterial or chloroplast counterparts, although the sequence identity or even similarity is very low. We attribute these findings to the different roles of these subunits in F1: The large alpha and beta subunits contribute to the catalytic centers of the enzyme, a function rendering them very sensitive to even minor changes. For the smaller delta and epsilon subunits it was sufficient to maintain a certain tertiary structure during evolution, with little emphasis on the conservation of particular amino acids.

  20. Selection and characterization of a candidate therapeutic bacteriophage that lyses the Escherichia coli O104:H4 strain from the 2011 outbreak in Germany.

    Directory of Open Access Journals (Sweden)

    Maia Merabishvili

    Full Text Available In 2011, a novel strain of O104:H4 Escherichia coli caused a serious outbreak of foodborne hemolytic uremic syndrome and bloody diarrhea in Germany. Antibiotics were of questionable use and 54 deaths occurred. Candidate therapeutic bacteriophages that efficiently lyse the E. coli O104:H4 outbreak strain could be selected rather easily from a phage bank or isolated from the environment. It is argued that phage therapy should be more considered as a potential armament against the growing threat of (resistant bacterial infections.

  1. Pathotyping and antibiotic resistance of porcine enterovirulent Escherichia coli strains from Switzerland (2014-2015).

    Science.gov (United States)

    Brand, P; Gobeli, S; Perreten, V

    2017-07-01

    A total of 131 porcine E. coli were isolated in 2014 and 2015 from the gut of 115 pigs raised in Switzerland and suffering from diarrhea. The isolates were tested for antibiotic resistance, serotypes, virulence factors and genetic diversity. Serotypes were assigned by agglutination tests and virulence genes were identified by polymerase chain reaction (PCR). Antibiotic resistance profile was determined by the measurement of the MIC of 14 antibiotics and by the detection of the corresponding genes using microarray and PCR approaches. Genetic diversity was determined by repetitive palindromic PCR (rep- PCR) revealing a heterogenous population. Half of the E. coli isolates possessing virulence factors could not be assigned to any of the 19 serotypes tested, but contained toxins and adhesins similarly to the sero-typable E. coli isolates. The most prevalent E. coli serotypes found were K88ac (18%), O139:K82 (6%), O141:K85ac (5%), O108:K`V189` (5%), O119:K`V113` (3%) and O157:K`V17` (2%). The combination of toxins EAST-1, STb and LT-I and adhesin F4 characterizing ETEC was the most frequent. The shigatoxin Stx2e (STEC) and intimin Eae (EPEC) were also detected, but less frequently. Seventy percent of the isolates were resistant to at least one antibiotic and 29% were resistant to more than 3 antibiotics. Isolates exhibited resistance to tetracycline (50%) associated to resistance genes tet(A), tet(B) and tet(C), sulfamethoxazole (49%) [sul1, sul2 and sul3], trimethoprim (34%) [dfr], nalidixic acid (29%), ampicillin (26%) [blaTEM-1], gentamicin (17%) [aac(3) -IIc, aac(3) -IVa and aac(3) -VIa], chloramphenicol (17%) [catAI and catAIII], and ciprofloxacin (8%) [mutations in GyrA (S83L) and ParC (S80I)]. All isolates were susceptible to 3rd generation cephalosporins, carbapenems, colistin and tigecycline. Pathogenic E. coli isolates from pigs in Switzerland could frequently not be assigned to a known serotype even if they contained diarrhea-causing virulence factors. They

  2. Carriage of extended-spectrum beta-lactamase-plasmids does not reduce fitness but enhances virulence in some strains of pandemic E. coli lineages

    Directory of Open Access Journals (Sweden)

    Katharina eSchaufler

    2016-03-01

    Full Text Available Pathogenic ESBL-producing E. coli lineages occur frequently worldwide, not only in a human health context but in animals and the environment, also in settings with low antimicrobial pressures. This study investigated the fitness costs of ESBL-plasmids and their influence on chromosomally encoded features associated with virulence, such as those involved in the planktonic and sessile behaviors of ST131 and ST648 E. coli. ESBL-plasmid-carrying wild-type E. coli strains, their corresponding ESBL-plasmid-cured variants (PCV, and complementary ESBL-carrying transformants were comparatively analyzed using growth curves, Omnilog® phenotype microarray (PM assays, macrocolony and biofilm formation, swimming motility, and RNA sequence analysis. Growth curves and PM results pointed towards similar growth and metabolic behaviors among the strains. Phenotypic differences in some strains were detected, including enhanced curli fimbriae and/or cellulose production as well as a reduced swimming capacity of some ESBL-carrying strains, as compared to their respective PCVs. RNA sequencing mostly confirmed the phenotypic results, suggesting that the chromosomally encoded csgD pathway is a key factor involved. These results contradict the hypothesis that ESBL-plasmid-carriage leads to a fitness loss in ESBL-carrying strains. Instead, the results indicate an influence of some ESBL-plasmids on chromosomally encoded features associated with virulence in some E. coli strains. In conclusion, apart from antibiotic resistance selective advantages, ESBL-plasmid-carriage may also lead to enhanced virulence or adaption to specific habitats in some strains of pandemic ESBL-producing E. coli lineages.

  3. Association of IL-8-inducing strains of diffusely adherent Escherichia coli with sporadic diarrheal patients with less than 5 years of age

    Directory of Open Access Journals (Sweden)

    Ismail Mustafa Meraz

    2007-02-01

    Full Text Available The role of diffusely adherent Escherichia coli (DAEC in diarrheal disease has been controversial. However, DAEC strains were recently implicated in diarrheal disease in developing countries. To clarify whether DAEC are prevalent among sporadic cases of diarrheal illness in Osaka City, Japan, E. coli strains isolated between July 1997 and March 2000 during diarrheagenic E. coli (DEC investigation were retrospectively examined. DAEC strains were recognized among 41 (4.4% of 924 patients and formed the biggest subgroup of DEC. Previously, we reported that some DAEC strains caused epithelial cells to secrete as much IL-8 as enteroaggregative E. coli strains did. In this study, we attempted to evaluate epidemiologically whether the ability of DAEC to induce IL-8 was involved in the pathogenesis. Relationship among patient age, symptoms, Afa adhesins, season and IL-8 induction were examined. The subgroup of DAEC that possessed Afa genes and/or induced a high level of IL-8 was significantly prevalent among patients age 1 to 4 years; however total DAEC was not significantly high among the children compared to other age group. IL-8 inducing DAEC seems to play a role in causing sporadic diarrheal illnesses, particularly in pediatric fields. Investigations highlighting the relationship between IL-8 induction and enteropathogenicity are clearly necessary to confirm the role of DAEC in infectious enteritis.

  4. Association of IL-8-inducing strains of diffusely adherent Escherichia coli with sporadic diarrheal patients with less than 5 years of age.

    Science.gov (United States)

    Meraz, Ismail Mustafa; Arikawa, Kentaro; Nakamura, Hiromi; Ogasawara, Jun; Hase, Atsushi; Nishikawa, Yoshikazu

    2007-02-01

    The role of diffusely adherent Escherichia coli (DAEC) in diarrheal disease has been controversial. However, DAEC strains were recently implicated in diarrheal disease in developing countries. To clarify whether DAEC are prevalent among sporadic cases of diarrheal illness in Osaka City, Japan, E. coli strains isolated between July 1997 and March 2000 during diarrheagenic E. coli (DEC) investigation were retrospectively examined. DAEC strains were recognized among 41 (4.4%) of 924 patients and formed the biggest subgroup of DEC. Previously, we reported that some DAEC strains caused epithelial cells to secrete as much IL-8 as enteroaggregative E. coli strains did. In this study, we attempted to evaluate epidemiologically whether the ability of DAEC to induce IL-8 was involved in the pathogenesis. Relationship among patient age, symptoms, Afa adhesins, season and IL-8 induction were examined. The subgroup of DAEC that possessed Afa genes and/or induced a high level of IL-8 was significantly prevalent among patients age 1 to 4 years; however total DAEC was not significantly high among the children compared to other age group. IL-8 inducing DAEC seems to play a role in causing sporadic diarrheal illnesses, particularly in pediatric fields. Investigations highlighting the relationship between IL-8 induction and enteropathogenicity are clearly necessary to confirm the role of DAEC in infectious enteritis.

  5. A novel multidrug resistance plasmid isolated from an Escherichia coli strain resistant to aminoglycosides.

    Science.gov (United States)

    Sun, Hui; Li, Shasha; Xie, Zhijing; Yang, Fangfang; Sun, Yani; Zhu, Yanli; Zhao, Xiaomin; Jiang, Shijin

    2012-07-01

    Previous studies have reported several different plasmids that confer multidrug resistance (MDR) including resistance to aminoglycosides. In this study, we investigated the aminoglycoside resistance patterns for 224 Escherichia coli isolates from diseased chickens and ducks in China, characterized a novel MDR plasmid, and collected prevalence data on similar resistance plasmids. Antibiotic susceptibilities were determined using disc diffusion and the microdilution method. The plasmid pXZ was analysed by restriction fragment length polymorphism (RFLP) with EcoRI and SalI, and sequenced. The prevalence of similar resistance plasmids was assessed by multiplex PCR and by RFLP analysis. Among the 224 E. coli isolates, 189 (84.4%) were resistant to streptomycin, 125 (55.8%) were resistant to kanamycin, 116 (51.8%) were resistant to gentamicin, 106 (47.3%) were resistant to neomycin and 98 (43.8%) were resistant to amikacin. Among the 224 E. coli isolates, 17 contained a plasmid with the MDR-encoding region of pXZ, which showed high-level resistance to aminoglycosides (MICs of gentamicin and amikacin ≥ 512 mg/L). The plasmid pXZ was digested into five fragments by EcoRI and six fragments by SalI. The plasmid pXZ was a circular DNA molecule of 76635 bp with a 51.65% guanine + cytosine content and included four resistance genes (rmtB, fosA3, bla(TEM-1) and bla(CTX-M-24)). A novel MDR plasmid, pXZ, harbouring four resistance genes (rmtB, fosA3, bla(TEM-1) and bla(CTX-M)) was identified. To our knowledge, this is the first report of an aminoglycoside resistance plasmid harbouring the fosA3 gene.

  6. Evaluation of seamless ligation cloning extract preparation methods from an Escherichia coli laboratory strain.

    Science.gov (United States)

    Okegawa, Yuki; Motohashi, Ken

    2015-10-01

    Seamless ligation cloning extract (SLiCE) is a simple and efficient method for DNA cloning without the use of restriction enzymes. Instead, SLiCE uses homologous recombination activities from Escherichia coli cell lysates. To date, SLiCE preparation has been performed using an expensive commercially available lytic reagent. To expand the utility of the SLiCE method, we evaluated different methods for SLiCE preparation that avoid using this reagent. Consequently, cell extracts prepared with buffers containing Triton X-100, which is a common and low-cost nonionic detergent, exhibited sufficient cloning activity for seamless gene incorporation into a vector.

  7. Apramycin treatment affects selection and spread of a multidrug-resistant Escherichia coli strain able to colonize the human gut in the intestinal microbiota of pigs

    DEFF Research Database (Denmark)

    Herrero-Fresno, Ana; Zachariasen, Camilla; Hansen, Monica Hegstad;

    2016-01-01

    The effect of apramycin treatment on transfer and selection of an Escherichia coli strain (E. coli 912) in the intestine of pigs was analyzed through an in vivo experiment. The strain was sequenced and assigned to the sequence type ST101 and serotype O11. It carried resistance genes to apramycin...... of treatment, and apramycin treatment resulted in significantly higher counts compared to the non-treated group. This represents the first demonstration of how antimicrobial treatment affects spread of resistant bacteria in pig production. The use of apramycin may lead to enhanced spread of gentamicin-resistant......-treated (pen 3), along with a non-inoculated control group (pen 1). Two pigs of pen 2 and 3 were inoculated intragastrically with a rifampicin resistant variant of the strain. Apramycin treatment in pen 2 was initiated immediately after inoculation. Strain colonization was assessed in the feces from all pigs...

  8. Improved method for expression and isolation of the Mycoplasma hominis arginine deiminase from the recombinant strain of Escherichia coli.

    Science.gov (United States)

    Fayura, Lyubov R; Boretsky, Yuriy R; Pynyaha, Yuriy V; Wheatley, Denys N; Sibirny, Andriy A

    2013-09-20

    Arginine deiminase is a promising anticancer drug active against melanoma, hepatocarcinoma and other tumors. Recombinant strains of Escherichia coli that express arginine deiminase from pathogenic bacteria Mycoplasma have been developed. However, production costs of heterologous arginine deiminase are high due to use of an expensive inducer and extraction buffer, as well as using diluted culture for enzyme induction. We report on a new advanced protocol for Mycoplasma hominis arginine deiminase expression, extraction and renaturation. The main improvements include manipulation with dense suspensions of E. coli, use of lactose instead of isopropyl β-D-1-thiogalactopyranoside as an inducer and a cheaper but not less efficient buffer for solubilization of arginine deiminase inclusion bodies. In addition, supplementation of the storage culture medium with glucose and substrate (arginine) significantly stabilized the recombinant arginine deiminase producer. Homogenous preparations of recombinant arginine deiminase were obtained using anion-exchange and hydrophobic chromatography. The purified enzyme retained a specific activity of 30-34 U/mg for 12 months when stored at 4°C in 20 mM sodium phosphate buffer pH 7.2 containing 1 M NaCl.

  9. The flagellin hypervariable region is a potential flagella display domain in probiotic Escherichia coli strain Nissle 1917.

    Science.gov (United States)

    Yang, Ying; Yang, Yi; Ou, Bingming; Xia, Pengpeng; Zhou, Mingxu; Li, Luan; Zhu, Guoqiang

    2016-09-01

    The most studied probiotic, Escherichia coli strain Nissle 1917 (EcN) possesses flagella of serotype H1. To explore the potential to use EcN flagellin in flagella display applications, we investigated the effect of deleting amino acids in the hypervariable region of flagellin on EcNc (EcN cured of its two cryptic plasmids pMUT1 and pMUT2). Two EcNc flagellin isogenic mutants with deletions of amino acid residual from 277 to 286 and from 287 to 296 in the hypervariable domain were constructed. Both mutants were flagellated, adherent to IPEC-J2 cells, and colonized BALB/c mice. These hypervariable regions may have future utility in the display of heterologous epitopes.

  10. Prevalence of adhesive genes among uropathogenic Escherichia coli strains isolated from patients with urinary tract infection in Mangalore

    Directory of Open Access Journals (Sweden)

    A V Shetty

    2014-01-01

    Full Text Available The study was carried out to detect the adhesive genes pap (pyelonephritis associated pili, sfa (S fimbrial adhesin and afa (afimbrial adhesin from Escherichia coli strains isolated in patients diagnosed with urinary tract infection (UTI. A total of 23% of the isolates were positive for pap, sfa and afa genes with a prevalence of 60.87% (14/23, 39.1% (9/23 and 39.1% (9/23, respectively. Prevalence of multiple adhesive genes was 8.7% (2/23 for pap and afa, 30.43% (7/23 for pap and sfa. Significant numbers of isolates were positive for Congo red binding (80% and haemolysin production 60%. The prevalence of multiple adhesive genes indicate the potential to adhere and subsequently cause a systemic infection among UTI patients.

  11. Biological and genetic characteristics of uropathogenic Escherichia coli strains Características biológicas e estrutura clonal em amostras uropatogênicas de Escherichia coli

    Directory of Open Access Journals (Sweden)

    Wanderley Dias da SILVEIRA

    2001-12-01

    Full Text Available The aim of the present study was to determine biological characteristics such as expression of fimbriae, Congo red binding, production of hemolysin and aerobactin, adhesion to HeLa and uroepithelial cells and invasion of HeLa cells by Escherichia coli isolates obtained from patients showing clinical signs of urinary tract infection (UTI. Also, the presence of genes (apa, afa, spa for fimbria expression and cytotoxic necrotizing factors (CNF1, CNF2 was assayed using specific primers in PCR. The data obtained were compared with the clonal relationships obtained by analysis of multilocus enzyme electrophoresis (MLEE, restriction fragment length polymorphism (RFLP of the rDNA (ribotyping and enterobacterial repetitive intergenic consensus-PCR (ERIC-PCR. All isolates but one presented a combination of at least two of the characteristics studied, a fact suggesting the presence of pathogenicity islands (PAIs. Diffuse adherence type to HeLa cells was observed to occur in most of the strains, but adhesion to uroepithelial cells seems to be a more reliable test to verify pathogenicity. Although four strains seemed to be able to invade HeLa cells when assayed by light microscopy, electron microscopy studies demonstrated that these strains were not invasive. MLEE, RFLP and ERIC-PCR were able to group the isolates differently into main clusters that were not correlated with the presence of pathogenic traits.O objetivo deste trabalho foi estudar características biológicas tais como a expressão de fímbrias e adesinas, capacidade de absorção do corante Vermelho Congo, produção de hemolisina e aerobactina, adesão e invasão a células HeLa e adesão a células do epitélio urinário em amostras de Eschericia coli isoladas de pacientes com sinais clínicos de infecção do trato urinário (UTI. A presença dos genes responsáveis pela expressão de fímbrias (apa, afa e spa e das Citotoxinas Necrotizantes CNF1, CNF2 foi avaliada por PCR. Esses dados

  12. Purification and characterization of lipopolysaccharides from six strains of non-O157 Shiga toxin-producing Escherichia coli.

    Science.gov (United States)

    Stromberg, Loreen R; Stromberg, Zachary R; Banisadr, Afsheen; Graves, Steven W; Moxley, Rodney A; Mukundan, Harshini

    2015-09-01

    Certain Shiga toxin-producing Escherichia coli (STEC) are virulent human pathogens that are most often acquired through contaminated food. The United States Department of Agriculture, Food Safety and Inspection Service has declared several serogroups of STEC as adulterants in non-intact raw beef products. Hence, sensitive and specific tests for the detection of these STEC are a necessity for implementation in food safety programs. E. coli serogroups are identified by their respective O-antigen moiety on the lipopolysaccharide (LPS) macromolecule. We propose that the development of O-antigen-specific immunological assays can facilitate simple and rapid discriminatory detection of STEC in beef. However, the resources (antigens and antibodies) required for such development are not readily available. To overcome this, we extracted and characterized LPS and O-antigen from six STEC strains. Using hot phenol extraction, we isolated the LPS component from each strain and purified it using a series of steps to eliminate proteins, nucleic acids, and lipid A antigens. Antigens and crude LPS extracts were characterized using gel electrophoresis, immunoblotting, and modified Western blotting with commercially available antibodies, thus assessing the serogroup specificity and sensitivity of available ligands as well. The results indicate that, while many commercially available antibodies bind LPS, their activities and specificities are highly variable, and often not as specific as those required for serogroup discrimination. This variability could be minimized by the production of antibodies specific for the O-antigen. Additionally, the antigens generated from this study provide a source of characterized LPS and O-antigen standards for six serogroups of STEC. Published by Elsevier B.V.

  13. Epidemiology of Escherichia coli, Klebsiella species, and Proteus mirabilis strains producing extended-spectrum β-lactamases from clinical samples in the Kinki Region of Japan.

    Science.gov (United States)

    Nakamura, Tatsuya; Komatsu, Masaru; Yamasaki, Katsutoshi; Fukuda, Saori; Miyamoto, Yugo; Higuchi, Takeshi; Ono, Tamotsu; Nishio, Hisaaki; Sueyoshi, Noriyuki; Kida, Kenji; Satoh, Kaori; Toda, Hirofumi; Toyokawa, Masahiro; Nishi, Isao; Sakamoto, Masako; Akagi, Masahiro; Nakai, Isako; Kofuku, Tomomi; Orita, Tamaki; Wada, Yasunao; Zikimoto, Takuya; Koike, Chihiro; Kinoshita, Shohiro; Hirai, Itaru; Takahashi, Hakuo; Matsuura, Nariaki; Yamamoto, Yoshimasa

    2012-04-01

    In the present study, nonduplicate, clinical isolates of extended-spectrum β-lactamase (ESBL)-producing Escherichia coli, Klebsiella spp, and Proteus mirabilis were collected during a 10-year period from 2000 to 2009 at several hospitals in the Kinki region, Japan. The detection rate of E coli markedly increased from 0.24% to 7.25%. The detection rate of Klebsiella pneumoniae increased from 0% to 2.44% and that of P mirabilis from 6.97% to 12.85%. The most frequently detected genotypes were the CTX-M9 group for E coli, the CTX-M2 group for K pneumoniae, and the CTX-M2 group for P mirabilis. E coli clone O25:H4-ST131 producing CTX-M-15, which is spreading worldwide, was first detected in 2007. The most common replicon type of E coli was the IncF type, particularly FIB, detected in 466 strains (69.7%). Of the K pneumoniae strains, 47 (55.3%) were of the IncN type; 77 P mirabilis strains (96.3%) were of the IncT type. In the future, the surveillance of various resistant bacteria, mainly ESBL-producing Enterobacteriaceae, should be expanded to prevent their spread.

  14. Genetic Virulence Profile of Enteroaggregative Escherichia coli Strains Isolated from Danish Children with Either Acute or Persistent Diarrhea

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    Betina Hebbelstrup Jensen

    2017-05-01

    Full Text Available Enteroaggregative Escherichia coli (EAEC is frequently found in diarrheal stools worldwide. It has been associated with persistent diarrhea, weight loss, and failure to thrive in children living in developing countries. A number of important EAEC virulence genes are identified; however, their roles in acute and persistent diarrhea have not been previously investigated. The aim of this study was to identify specific EAEC virulence genes associated with duration and type of diarrhea in Danish children. We aimed to improve the current diagnostics of EAEC and enable targeting of strains with an expected severe disease course. Questionnaires answered by parents provided information regarding duration of diarrhea and presence of blood or mucus. A total of 295 EAEC strains were collected from children with acute (≤7 days and persistent diarrhea (≥14 days and were compared by using multiplex PCR targeting the genes sat, sepA, pic, sigA, pet, astA, aatA, aggR, aaiC, aap, agg3/4C, ORF3, aafA, aggA, agg3A, agg4A, and agg5A. Furthermore, the distribution of EAEC genes in strains collected from cases of bloody, mucoid, and watery diarrhea was investigated. The classification and regression tree analysis (CART was applied to investigate the relationship between EAEC virulence genes and diarrheal duration and type. Persistent diarrhea was associated with strains lacking the pic gene (p = 0.002 and with the combination of the genes pic, sat, and absence of the aggA gene (p = 0.05. Prolonged diarrhea was associated with the combination of the genes aatA and astA (p = 0.03. Non-mucoid diarrhea was associated with strains lacking the aatA gene (p = 0.004. Acute diarrhea was associated with the genes aggR, aap, and aggA by individual odds ratios. Resistance toward gentamicin and ciprofloxacin was observed in 7.5 and 3% of strains, respectively. Multi-drug resistance was observed in 38% of strains. Genetic host factors have been associated with an increased risk

  15. Nickel Promotes Biofilm Formation by Escherichia coli K-12 Strains That Produce Curli▿

    Science.gov (United States)

    Perrin, Claire; Briandet, Romain; Jubelin, Gregory; Lejeune, Philippe; Mandrand-Berthelot, Marie-Andrée; Rodrigue, Agnès; Dorel, Corinne

    2009-01-01

    The survival of bacteria exposed to toxic compounds is a multifactorial phenomenon, involving well-known molecular mechanisms of resistance but also less-well-understood mechanisms of tolerance that need to be clarified. In particular, the contribution of biofilm formation to survival in the presence of toxic compounds, such as nickel, was investigated in this study. We found that a subinhibitory concentration of nickel leads Escherichia coli bacteria to change their lifestyle, developing biofilm structures rather than growing as free-floating cells. Interestingly, whereas nickel and magnesium both alter the global cell surface charge, only nickel promotes biofilm formation in our system. Genetic evidence indicates that biofilm formation induced by nickel is mediated by the transcriptional induction of the adhesive curli-encoding genes. Biofilm formation induced by nickel does not rely on efflux mechanisms using the RcnA pump, as these require a higher concentration of nickel to be activated. Our results demonstrate that the nickel-induced biofilm formation in E. coli is an adaptational process, occurring through a transcriptional effect on genes coding for adherence structures. The biofilm lifestyle is obviously a selective advantage in the presence of nickel, but the means by which it improves bacterial survival needs to be investigated. PMID:19168650

  16. Nickel promotes biofilm formation by Escherichia coli K-12 strains that produce curli.

    Science.gov (United States)

    Perrin, Claire; Briandet, Romain; Jubelin, Gregory; Lejeune, Philippe; Mandrand-Berthelot, Marie-Andrée; Rodrigue, Agnès; Dorel, Corinne

    2009-03-01

    The survival of bacteria exposed to toxic compounds is a multifactorial phenomenon, involving well-known molecular mechanisms of resistance but also less-well-understood mechanisms of tolerance that need to be clarified. In particular, the contribution of biofilm formation to survival in the presence of toxic compounds, such as nickel, was investigated in this study. We found that a subinhibitory concentration of nickel leads Escherichia coli bacteria to change their lifestyle, developing biofilm structures rather than growing as free-floating cells. Interestingly, whereas nickel and magnesium both alter the global cell surface charge, only nickel promotes biofilm formation in our system. Genetic evidence indicates that biofilm formation induced by nickel is mediated by the transcriptional induction of the adhesive curli-encoding genes. Biofilm formation induced by nickel does not rely on efflux mechanisms using the RcnA pump, as these require a higher concentration of nickel to be activated. Our results demonstrate that the nickel-induced biofilm formation in E. coli is an adaptational process, occurring through a transcriptional effect on genes coding for adherence structures. The biofilm lifestyle is obviously a selective advantage in the presence of nickel, but the means by which it improves bacterial survival needs to be investigated.

  17. Caffeic acid production enhancement by engineering a phenylalanine over-producing Escherichia coli strain.

    Science.gov (United States)

    Huang, Qin; Lin, Yuheng; Yan, Yajun

    2013-12-01

    Caffeic acid is a plant-specific phenylpropanoic acid with multiple health-improving effects reported, and its therapeutic derivatives have also been studied throughout the last decade. To meet its market need and achieve high-level production, microbial production of caffeic acid approaches have been developed in metabolically engineered Escherichia coli. In our previous work, we have established the first artificial pathway that realized de novo production of caffeic acid using E. coli endogenous 4-hydroxyphenylacetate 3-hydroxylase (4HP3H). In this work, we exploited the catalytic potential of 4HPA3H in the whole-cell bioconversion study and produced 3.82 g/L (461.12 mg/L/OD) caffeic acid from p-coumaric acid, a direct precursor. We further engineered a phenylalanine over-producer into a tyrosine over-producer and then introduced the artificial pathway. After adjusting the expression strategy and optimizing the inoculants timing, de novo production of caffeic acid reached 766.68 mg/L. Both results from the direct precursor and simple carbon sources represent the highest titers of caffeic acid from microbial production so far.

  18. Pulsed-field gel electrophoresis typing of Eschericia coli strains from samples collected before and after pivmecillinam or placebo treatment of uncomplicated community-acquired urinary tract infection in women

    DEFF Research Database (Denmark)

    Ejrnæs, K; Sandvang, D; Lundgren, Bettina

    2006-01-01

    The primary infecting Escherichia coli strains from 156 women with community-acquired uncomplicated urinary tract infection (UTI) randomized to pivmecillinam or placebo and the E. coli strains causing UTI at two follow-up visits were typed using pulsed-field gel electrophoresis (PFGE...

  19. Pulsed-field gel electrophoresis typing of Escherichia coli strains from samples collected before and after pivmecillinam or placebo treatment of uncomplicated community-acquired urinary tract infection in women

    DEFF Research Database (Denmark)

    Ejrnaes, Karen; Sandvang, Dorthe; Lundgren, Bettina

    2006-01-01

    The primary infecting Escherichia coli strains from 156 women with community-acquired uncomplicated urinary tract infection (UTI) randomized to pivmecillinam or placebo and the E. coli strains causing UTI at two follow-up visits were typed using pulsed-field gel electrophoresis (PFGE...

  20. Pulsed-field gel electrophoresis typing of Eschericia coli strains from samples collected before and after pivmecillinam or placebo treatment of uncomplicated community-acquired urinary tract infection in women

    DEFF Research Database (Denmark)

    Ejrnæs, K; Sandvang, D; Lundgren, Bettina

    2006-01-01

    ). The finding that the majority of UTIs at follow-up are caused by the primary infecting E. coli strain supports the theory of a vaginal and rectal reservoir but could also support the recent discovery that E. coli strains are able to persist in the bladder epithelium despite appropriate antibiotic treatment...

  1. Muc2基因沉默可降低益生菌抑制大肠杆菌黏附和侵袭的能力%Effect on Muc2 gene knockdown in Ht29 cells by CRISPR/Cas9 on probiotics-mediated inhibition of E.coli K1 adhesion and invasion

    Institute of Scientific and Technical Information of China (English)

    邱嘉文; 曹虹; 何肖龙; 张宝; 杜蕾; 曾庆; 李森; 熊欢欢; 龙敏; 罗军

    2016-01-01

    目的:建立抑制肠上皮细胞基因组中Muc2基因表达的CRISPR/Cas9系统并探索粘蛋白Muc2在鼠李糖乳杆菌GG株(LGG)抑制大肠杆菌K1(Escherichia coli, E.coli k1)株E44黏附和侵袭肠上皮中的作用机制。方法设计2个长20~25 bp的sgRNA分别靶向Muc2,合成sgRNA寡核苷酸序列并构建CRISPR表达载体,转染野生型人结肠癌Ht29细胞,蛋白免疫印迹法检测抑制效率及通过MTT法检测其细胞活力及生长情况后,竞争性排斥分析验证粘蛋白Muc2在益生菌抑制E44黏附侵袭肠上皮中的作用。结果目的sgRNA寡核苷酸双链成功插入酶切后的lenticrisprv2质粒载体中且序列正确;稳定抑制Muc2表达的细胞株筛选成功;Muc2基因沉默后,与空白对照组相比,其表达水平明显降低,抑制率可达81%(P<0.01);黏附侵袭实验中E44相对黏附率为72.23%(P<0.01),相对侵袭率为81.49%(P<0.01),益生菌LGG的抑制E44黏附和侵袭作用明显下调。结论Muc2基因下调明显降低益生菌抑制E44黏附和侵袭肠上皮细胞的能力,提示益生菌刺激Muc2表达上调可能是其强化加固肠粘膜屏障和拮抗致病菌功能的关键性机制之一,并可为肠道细菌性感染疾病的预防治疗提供了一个新手段。%Objective To investigate the effects of Lactobacillus rhamnosus GG (LGG) for inhibiting E.coli K1 (E44) adhesion and invasion of an intestinal epithelial cell model with Muc2 gene knockdown established using CRISPR-Cas9 system. Methods Two 20-25 bp sgRNAs targeting Muc2 were chemically synthesized to construct CRISPR expression vectors for transfection in wild-type human colonic cancer cell line Ht29. The efficiency of Muc2 knockdown was determined using Western blotting. After assessment of the viability and proliferation of the transfected cells with MTT assay, we evaluated the effects of the probiotics against E44 adhesion and invasion of the cells through a

  2. Antibiotic resistance in Escherichia coli strains isolated from Antarctic bird feces, water from inside a wastewater treatment plant, and seawater samples collected in the Antarctic Treaty area

    Science.gov (United States)

    Rabbia, Virginia; Bello-Toledo, Helia; Jiménez, Sebastián; Quezada, Mario; Domínguez, Mariana; Vergara, Luis; Gómez-Fuentes, Claudio; Calisto-Ulloa, Nancy; González-Acuña, Daniel; López, Juana; González-Rocha, Gerardo

    2016-06-01

    Antibiotic resistance is a problem of global concern and is frequently associated with human activity. Studying antibiotic resistance in bacteria isolated from pristine environments, such as Antarctica, extends our understanding of these fragile ecosystems. Escherichia coli strains, important fecal indicator bacteria, were isolated on the Fildes Peninsula (which has the strongest human influence in Antarctica), from seawater, bird droppings, and water samples from inside a local wastewater treatment plant. The strains were subjected to molecular typing with pulsed-field gel electrophoresis to determine their genetic relationships, and tested for antibiotic susceptibility with disk diffusion tests for several antibiotic families: β-lactams, quinolones, aminoglycosides, tetracyclines, phenicols, and trimethoprim-sulfonamide. The highest E. coli count in seawater samples was 2400 cfu/100 mL. Only strains isolated from seawater and the wastewater treatment plant showed any genetic relatedness between groups. Strains of both these groups were resistant to β-lactams, aminoglycosides, tetracycline, and trimethoprim-sulfonamide.In contrast, strains from bird feces were susceptible to all the antibiotics tested. We conclude that naturally occurring antibiotic resistance in E. coli strains isolated from Antarctic bird feces is rare and the bacterial antibiotic resistance found in seawater is probably associated with discharged treated wastewater originating from Fildes Peninsula treatment plants.

  3. Fatores de virulência presentes em amostras de Escherichia coli uropatogênicas - UPEC para suínos Virulence factors of uropathogenic Escherichia coli - UPEC strains for pigs

    Directory of Open Access Journals (Sweden)

    Benito Guimarães de Brito

    2004-04-01

    Full Text Available As infecções urinárias são freqüentes nos rebanhos suínos, sendo a principal causa de descarte e mortalidade de animais adultos. Apesar das características multifatoriais da doença o microrganismo freqüentemente isolado é a Escherichia coli. Vários fatores de virulência de Escherichia coli foram descritos em amostras uropatogênicas e permitem diferenciar cepas patogênicas de não patogênicas. Esta revisão tem por objetivo apresentar alguns tópicos relativos aos fatores de virulência presentes em amostras de E. coli uropatogênicas para suínos.Urinary tract infections occur frequently in pig herds urinary infection is the most significant cause of culling and mortality of adult animals. Despite the multifactorial nature of this condition, Escherichia coli is frequently isolated from diseased animals. Several virulence factors were described on uropathogenic strains and they can be used to distinguish isolates. The objective of the present review is to present some topics related to virulence factors present in swine uropathogenic E. coli strains.

  4. Genetic relatedness of commensal Escherichia coli from nursery pigs in intensive pig production in Denmark and molecular characterization of genetically different strains

    DEFF Research Database (Denmark)

    Herrero Fresno, Ana; Larsen, Inge; Olsen, John Elmerdahl

    2015-01-01

    AIMS: To determine the genetic relatedness and the presence of virulence and antibiotic resistance genes in commensal Escherichia coli from nursery pigs in Danish intensive production. METHODS AND RESULTS: The genetic diversity of 1000 E. coli strains randomly picked (N = 50 isolates) from cultured...... faecal samples (N = 4 pigs) from five intensive Danish pigs farms was analysed by repetitive extragenic palindromic-PCR (REP-PCR) and 42 unique REP-profiles were detected (similarity commensal E. coli......-producing genes were observed in 20 isolates. CONCLUSIONS: A low genetic diversity was found in commensal gut E. coli from nursery pigs in Denmark. No correlation was observed between REP-profiles, ST-types and resistance/virulence patterns. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study analysing...

  5. Whole genome sequencing of diverse Shiga toxin-producing and non-producing Escherichia coli strains reveals a variety of virulence and novel antibiotic resistance plasmids

    Science.gov (United States)

    The genomes of a diverse set of Shiga toxin-producing E. coli strains and the presence of 38 plasmids among all the isolates were determined. Among the novel plasmids found, there were eight that encoded resistance genes to antibiotics, including aminoglycosides, carbapenems, penicillins, cephalosp...

  6. Characterization and virulence potential of serogroup O113 Shiga toxin-producing Escherichia coli strains isolated from beef and cattle in the United States

    Science.gov (United States)

    Shiga toxin-producing Escherichia coli (STEC) of serotype O113:H21 have caused severe diseases but are unusual in that they do not produce the intimin protein required for adherence to intestinal epithelial cells. Strains of serogroup O113 are one of the most common STEC found in ground beef and be...

  7. Investigation of environmental factors on the prevalence of free bacteriophages against Shiga toxin-producing Escherichia coli strains in produce pre-harvest environment in Salinas, California

    Science.gov (United States)

    Investigation of environmental factors on the prevalence of free bacteriophages against Shiga toxin-producing Escherichia coli strains in produce pre-harvest environment in Salinas, California Yen-Te Liaoa, Irwin Quintelab, Kimberly Nguyena, Alexandra Salvadora, Michael Cooleya, and Vivian C.H. Wu*a...

  8. Preliminary Remarks Regarding the Prevalence of ESBL-Producing Strains of E. coli and K. Pneumoniae, Isolated from Cows with Clinical Endometritis

    Directory of Open Access Journals (Sweden)

    Cristina Ioana CRIVEI

    2017-05-01

    In this preliminary study, by phenotypic methods was confirmed a prevalence of 35.3% for the ESBL strains of E. coli and K. pneumoniae, which requires further research to confirm by molecular biology the identification of ESBL resistance genes, but also for the plasmids encoding these gene transmission.

  9. Stx1 prophage excision in Escherichia coli strain PA20 confers strong curli and biofilm formation by restoring native mlrA

    Science.gov (United States)

    Prophage insertions in Escherichia coli O157:H7 mlrA contribute to the low expression of curli fimbriae and biofilm observed in many clinical isolates. Varying levels of CsgD-dependent curli/biofilm expression are restored to strains bearing prophage insertions in mlrA by mutation of regulatory gene...

  10. Secretome Biomarkers for the Identification and Differentiation of Enterohemorrhagic and Enteropathogenic Escherichia coli Strains

    Science.gov (United States)

    2013-09-01

    flagellum hook AP_003849.1 DNA-binding transcriptional dual regulator EC O157:H7 Binding Transcription ND NP_288384.1 Flagellin EC O157:H7...capability is corroborated because prokaryotic organisms are arranged in hierarchical order; their common proteins increase as we move from strain to phyla

  11. A Commensal Gone Bad: Complete Genome Sequence of the Prototypical Enterotoxigenic Escherichia coli Strain H10407

    Science.gov (United States)

    2010-11-01

    tion (eorC). In addition, ETEC Hl0407 pETEC58 encodes a putative deoxy- cytidylatc deaminasc (pETEC58_0005). As me ntioned above, if a particula r...ami no acid similarity. qucnces and not the absence of particula r genes from these strains. T he d istributions and locations of the cargo genes

  12. Disinfectant and Antimicrobial Susceptibility Profiles of the Big Six Non-O157 Shiga Toxin-Producing Escherichia coli Strains from Food Animals and Humans.

    Science.gov (United States)

    Beier, Ross C; Franz, Eelco; Bono, James L; Mandrell, Robert E; Fratamico, Pina M; Callaway, Todd R; Andrews, Kathleen; Poole, Toni L; Crippen, Tawni L; Sheffield, Cynthia L; Anderson, Robin C; Nisbet, David J

    2016-08-01

    The disinfectant and antimicrobial susceptibility profiles of 138 non-O157 Shiga toxin-producing Escherichia coli strains (STECs) from food animals and humans were determined. Antimicrobial resistance (AMR) was moderate (39.1% of strains) in response to 15 antimicrobial agents. Animal strains had a lower AMR prevalence (35.6%) than did human strains (43.9%) but a higher prevalence of the resistance profile GEN-KAN-TET. A decreasing prevalence of AMR was found among animal strains from serogroups O45 > O145 > O121 > O111 > O26 > O103 and among human strains from serogroups O145 > O103 > O26 > O111 > O121 > O45. One animal strain from serogroups O121 and O145 and one human strain from serogroup O26 had extensive drug resistance. A high prevalence of AMR in animal O45 and O121 strains and no resistance or a low prevalence of resistance in human strains from these serogroups suggests a source other than food animals for human exposure to these strains. Among the 24 disinfectants evaluated, all strains were susceptible to triclosan. Animal strains had a higher prevalence of resistance to chlorhexidine than did human strains. Both animal and human strains had a similar low prevalence of low-level benzalkonium chloride resistance, and animal and human strains had similar susceptibility profiles for most other disinfectants. Benzyldimethylammonium chlorides and C10AC were the primary active components in disinfectants DC&R and P-128, respectively, against non-O157 STECs. A disinfectant FS512 MIC ≥ 8 μg/ml was more prevalent among animal O121 strains (61.5%) than among human O121 strains (25%), which may also suggest a source of human exposure to STEC O121 other than food animals. Bacterial inhibition was not dependent solely on pH but was correlated with the presence of dissociated organic acid species and some undissociated acids.

  13. No evidence of the Shiga toxin-producing E. coli O104:H4 outbreak strain or enteroaggregative E. coli (EAEC found in cattle faeces in northern Germany, the hotspot of the 2011 HUS outbreak area

    Directory of Open Access Journals (Sweden)

    Wieler Lothar H

    2011-11-01

    Full Text Available Abstract Background Ruminants, in particular bovines, are the primary reservoir of Shiga toxin-producing E. coli (STEC, but whole genome analyses of the current German ESBL-producing O104:H4 outbreak strain of sequence type (ST 678 showed this strain to be highly similar to enteroaggregative E. coli (EAEC. Strains of the EAEC pathotype are basically adapted to the human host. To clarify whether in contrast to this paradigm, the O104:H4 outbreak strain and/or EAEC may also be able to colonize ruminants, we screened a total of 2.000 colonies from faecal samples of 100 cattle from 34 different farms - all located in the HUS outbreak region of Northern Germany - for genes associated with the O104:H4 HUS outbreak strain (stx2, terD, rfbO104, fliCH4, STEC (stx1, stx2, escV, EAEC (pAA, aggR, astA, and ESBL-production (blaCTX-M, blaTEM, blaSHV. Results The faecal samples contained neither the HUS outbreak strain nor any EAEC. As the current outbreak strain belongs to ST678 and displays an en-teroaggregative and ESBL-producing phenotype, we additionally screened selected strains for ST678 as well as the aggregative adhesion pattern in HEp-2 cells. However, we were unable to find any strains belonging to ST678 or showing an aggregative adhesion pattern. A high percentage of animals (28% shed STEC, corroborating previous knowl-edge and thereby proving the validity of our study. One of the STEC also harboured the LEE pathogenicity island. In addition, eleven animals shed ESBL-producing E. coli. Conclusions While we are aware of the limitations of our survey, our data support the theory, that, in contrast to other Shiga-toxin producing E. coli, cattle are not the reservoir for the O104:H4 outbreak strain or other EAEC, but that the outbreak strain seems to be adapted to humans or might have yet another reservoir, raising new questions about the epidemiology of STEC O104:H4.

  14. Strain differences in fitness of Escherichia coli O157:H7 to resist protozoan predation and survival in soil.

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    Subbarao V Ravva

    Full Text Available Escherichia coli O157:H7 (EcO157 associated with the 2006 spinach outbreak appears to have persisted as the organism was isolated, three months after the outbreak, from environmental samples in the produce production areas of the central coast of California. Survival in harsh environments may be linked to the inherent fitness characteristics of EcO157. This study evaluated the comparative fitness of outbreak-related clinical and environmental strains to resist protozoan predation and survive in soil from a spinach field in the general vicinity of isolation of strains genetically indistinguishable from the 2006 outbreak strains. Environmental strains from soil and feral pig feces survived longer (11 to 35 days for 90% decreases, D-value with Vorticella microstoma and Colpoda aspera, isolated previously from dairy wastewater; these D-values correlated (P<0.05 negatively with protozoan growth. Similarly, strains from cow feces, feral pig feces, and bagged spinach survived significantly longer in soil compared to clinical isolates indistinguishable by 11-loci multi-locus variable-number tandem-repeat analysis. The curli-positive (C+ phenotype, a fitness trait linked with attachment in ruminant and human gut, decreased after exposure to protozoa, and in soils only C- cells remained after 7 days. The C+ phenotype correlated negatively with D-values of EcO157 exposed to soil (rs = -0.683; P = 0.036, Vorticella (rs = -0.465; P = 0.05 or Colpoda (rs = -0.750; P = 0.0001. In contrast, protozoan growth correlated positively with C+ phenotype (Vorticella, rs = 0.730, P = 0.0004; Colpoda, rs = 0.625, P = 0.006 suggesting a preference for consumption of C+ cells, although they grew on C- strains also. We speculate that the C- phenotype is a selective trait for survival and possibly transport of the pathogen in soil and water environments.

  15. Artificial biosynthesis of phenylpropanoic acids in a tyrosine overproducing Escherichia coli strain

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    Kang Sun-Young

    2012-12-01

    Full Text Available Abstract Background The phenylpropanoid metabolites are an extremely diverse group of natural products biosynthesized by plants, fungi, and bacteria. Although these compounds are widely used in human health care and nutrition services, their availability is limited by regional variations, and isolation of single compounds from plants is often difficult. Recent advances in synthetic biology and metabolic engineering have enabled artificial production of plant secondary metabolites in microorganisms. Results We develop an Escherichia coli system containing an artificial biosynthetic pathway that yields phenylpropanoic acids, such as 4-coumaric acid, caffeic acid, and ferulic acid, from simple carbon sources. These artificial biosynthetic pathways contained a codon-optimized tal gene that improved the productivity of 4-coumaric acid and ferulic acid, but not caffeic acid in a minimal salt medium. These heterologous pathways extended in E. coli that had biosynthesis machinery overproducing tyrosine. Finally, the titers of 4-coumaric acid, caffeic acid, and ferulic acid reached 974 mg/L, 150 mg/L, and 196 mg/L, respectively, in shake flasks after 36-hour cultivation. Conclusions We achieved one gram per liter scale production of 4-coumaric acid. In addition, maximum titers of 150 mg/L of caffeic acid and 196 mg/L of ferulic acid were achieved. Phenylpropanoic acids, such as 4-coumaric acid, caffeic acid, and ferulic acid, have a great potential for pharmaceutical applications and food ingredients. This work forms a basis for further improvement in production and opens the possibility of microbial synthesis of more complex plant secondary metabolites derived from phenylpropanoic acids.

  16. Nosocomial Intravascular Catheter Infections with Extended-spectrum Beta-lactamase-producing Escherichia coli in Calves after Strain Introduction from a Commercial Herd.

    Science.gov (United States)

    Pardon, B; Smet, A; Butaye, P; Argudín, M A; Valgaeren, B; Catry, B; Haesebrouck, F; Deprez, P

    2017-02-01

    An outbreak of intravascular catheter-related infections by extended-spectrum β-lactamase (ESBL)-producing Escherichia coli in calves in an animal teaching hospital is reported. Pulsed-field gel electrophoresis was used for strain typing to determine the origin and dissemination of these strains. All 19 strains harboured the blaCTX-M-14, and six strains also overexpressed their chromosomal AmpC gene. Evidence on the introduction of the strain from a beef herd, experiencing neonatal diarrhoea and increased mortality, to the clinic through admission of diarrhoeic calves was provided. Strains isolated from phlebitis cases from other herds up to 5 months later showed a high similarity with the initial strain, suggesting that the strain had become nosocomial. The catheter infections with ESBL/AmpC-producing E. coli resulted in a prolonged hospitalization, increased anti-microbial use and mortality. This report points towards the potential dangers of the emergence of ESBL/AmpC-producing bacteria in susceptible food animals and warns farmers and veterinarians for the facility by which they are introduced into another environment. © 2015 Blackwell Verlag GmbH.

  17. Carriage of stx2a differentiates clinical and bovine-biased strains of Escherichia coli O157.

    Directory of Open Access Journals (Sweden)

    Smriti Shringi

    Full Text Available BACKGROUND: Shiga toxin (Stx are cardinal virulence factors of enterohemorrhagic E. coli O157:H7 (EHEC O157. The gene content and genomic insertion sites of Stx-associated bacteriophages differentiate clinical genotypes of EHEC O157 (CG, typical of clinical isolates from bovine-biased genotypes (BBG, rarely identified among clinical isolates. This project was designed to identify bacteriophage-mediated differences that may affect the virulence of CG and BBG. METHODS: Stx-associated bacteriophage differences were identified by whole genome optical scans and characterized among >400 EHEC O157 clinical and cattle isolates by PCR. RESULTS: Optical restriction maps of BBG strains consistently differed from those of CG strains only in the chromosomal insertion sites of Stx2-associated bacteriophages. Multiplex PCRs (stx1, stx2a, and stx2c as well as Stx-associated bacteriophage-chromosomal insertion site junctions revealed four CG and three BBG that accounted for >90% of isolates. All BBG contained stx2c and Stx2c-associated bacteriophage-sbcB junctions. All CG contained stx2a and Stx2a-associated bacteriophage junctions in wrbA or argW. CONCLUSIONS: Presence or absence of stx2a (or another product encoded by the Stx2a-associated bacteriophage is a parsimonious explanation for differential virulence of BBG and CG, as reflected in the distributions of these genotypes in humans and in the cattle reservoir.

  18. Genetically similar strains of Escherichia coli O157:H7 isolated from sheep, cattle and human patients

    Directory of Open Access Journals (Sweden)

    Söderlund Robert

    2012-10-01

    Full Text Available Abstract Background Comparatively little is known about the prevalence or the molecular characteristics of the zoonotic pathogen E. coli O157:H7 in the sheep reservoir. To investigate this and determine the host specificity of subclones of the bacterium, we have conducted a slaughterhouse prevalence study in sheep and compared the collected isolates to O157:H7 previously isolated from cattle and human patients. Results Verotoxin-producing O157:H7 was found in 11/597 (1.8% of samples from sheep in Swedish slaughterhouses, 9/492 faecal (1.8% and 2/105 ear samples (1.9%. All positive sheep were eaeA, hlyA, cdtV-B, vtx1, and partial sequencing of vtx2. The observed profiles were similar to those of cattle strains investigated previously. Conclusions The same pathogenic subtypes of VTEC O157:H7, including the highly virulent clade 8, appear to be present in both sheep and cattle in Sweden, suggesting strains can circulate freely between ruminant reservoirs.

  19. Effects of antibiotics on Shiga toxin 2 production and bacteriophage induction by epidemic Escherichia coli O104:H4 strain.

    Science.gov (United States)

    Bielaszewska, Martina; Idelevich, Evgeny A; Zhang, Wenlan; Bauwens, Andreas; Schaumburg, Frieder; Mellmann, Alexander; Peters, Georg; Karch, Helge

    2012-06-01

    The role of antibiotics in treatment of enterohemorrhagic Escherichia coli (EHEC) infections is controversial because of concerns about triggering hemolytic-uremic syndrome (HUS) by increasing Shiga toxin (Stx) production. During the recent large EHEC O104:H4 outbreak, antibiotic therapy was indicated for some patients. We tested a diverse panel of antibiotics to which the outbreak strain is susceptible to interrogate the effects of subinhibitory antibiotic concentrations on induction of stx(2)-harboring bacteriophages, stx(2) transcription, and Stx2 production in this emerging pathogen. Ciprofloxacin significantly increased stx(2)-harboring phage induction and Stx2 production in outbreak isolates (P values of 0.1) and chloramphenicol, meropenem, azithromycin, rifaximin, and tigecycline significantly decreased them (P ≤ 0.05). Ciprofloxacin and chloramphenicol significantly upregulated and downregulated stx(2) transcription, respectively (P antibiotics had insignificant effects (P > 0.1). Meropenem, azithromycin, and rifaximin, which were used for necessary therapeutic or prophylactic interventions during the EHEC O104:H4 outbreak, as well as tigecycline, neither induced stx(2)-harboring phages nor increased stx(2) transcription or Stx2 production in the outbreak strain. These antibiotics might represent therapeutic options for patients with EHEC O104:H4 infection if antibiotic treatment is inevitable. We await further analysis of the epidemic to determine if usage of these agents was associated with an altered risk of developing HUS.

  20. Pulsed-field gel electrophoresis typing of Eschericia coli strains from samples collected before and after pivmecillinam or placebo treatment of uncomplicated community-acquired urinary tract infection in women

    DEFF Research Database (Denmark)

    Ejrnæs, K; Sandvang, D; Lundgren, Bettina

    2006-01-01

    The primary infecting Escherichia coli strains from 156 women with community-acquired uncomplicated urinary tract infection (UTI) randomized to pivmecillinam or placebo and the E. coli strains causing UTI at two follow-up visits were typed using pulsed-field gel electrophoresis (PFGE......). In the pivmecillinam treatment group PFGE showed that among patients having a negative urine culture at the first follow-up 77% (46/60) had a relapse with the primary infecting E. coli strain and 23% (14/60) had reinfection with a new E. coli strain at the second follow-up. Among patients having E. coli at the first...... follow-up PFGE showed that 80% (32/40) had persistence with the primary infecting E. coli strain, 15% (6/40) had reinfection with a new E. coli strain, and 5% (2/40) had different E. coli strains at the two follow-up visits (one had reinfection followed by relapse, and the other had persistence followed...

  1. Removal of Antibiotic Resistance of Live Vaccine Strain Escherichia coli MM-3 and Evaluation of the Immunogenicity of the New Strain

    Institute of Scientific and Technical Information of China (English)

    Sheng-Ling YUAN; Peng WANG; Hao-Xia TAO; Xiang-Xin LIU; Yan-Chun WANG; De-Wen ZHAN; Chun-Jie LIU; Zhao-Shan ZHANG

    2006-01-01

    MM-3 was a live vaccine strain candidate for protecting neonatal piglets from diarrhea.Designed in the 1980s, a high degree of protection from colibacillosis was afforded to piglets in a challenge study and field trials. However MM-3 had a drawback of carrying the antibiotic resistance gene (chloramphenicol acetyltransferase gene, cat). The introduction of a host-plasmid balanced lethal system into the vaccine was a good idea to solve the problem. The λ-Red recombination system was adopted in this study to realize the replacement of cat by aspartate-semialdehyde dehydrogenase gene (asd) in the plasmid pMM085. The new plasmid named pMMASD was introduced into an Escherichia coli strain χ6097 and Salmonella typhimurium χ4072 where the asd gene had been knocked out in their chromosomes. Cultured in an Erlenmeyer flask,expression levels of two antigens K88ac fimbriae and heat-labile enterotoxin B subunit (LTB) in cell lysate were similar among MM-3, χ4072(pMMASD) and χ6097(pMMASD). However, χ4072(pMMASD) possessed the more effective secretion mechanism to transport LTB enterotoxin into culture liquid. The relatively higher stability of pMMASD in Salmonella typhimurium χ4072 than that of pMM085 in MM-3 was determined both in vitro in the absence of selective pressure, and in vivo following oral inoculation. Oral immunization of BALB/c mice with χ4072(pMMASD) or χ6097(pMMASD) was sufficient to elicit IgA responses in mucosal tissues as well as systemic IgG antibody responses to the K88 fimbriae, while MM-3 failed to elicit specific antibody responses to K88 fimbriae in mucosal tissues. Among three live strains, only χ4072(pMMASD)could develop strong humoral responses against LTB enterotoxin. The results suggest that χ4072(pMMASD)is expected to be a promising live vaccine strain.

  2. Prevalence of adhesin and toxin genes in E. coli strains isolated from diarrheic and non-diarrheic pigs from smallholder herds in northern and eastern Uganda.

    Science.gov (United States)

    Ikwap, Kokas; Larsson, Jenny; Jacobson, Magdalena; Owiny, David Okello; Nasinyama, George William; Nabukenya, Immaculate; Mattsson, Sigbrit; Aspan, Anna; Erume, Joseph

    2016-08-05

    Enterotoxigenic E. coli (ETEC) significantly contribute to diarrhea in piglets and weaners. The smallholder pig producers in Uganda identified diarrhea as one of the major problems especially in piglets. The aim of this study was to; i) characterize the virulence factors of E. coli strains isolated from diarrheic and non-diarrheic suckling piglets and weaners from smallholder herds in northern and eastern Uganda and ii) identify and describe the post-mortem picture of ETEC infection in severely diarrheic piglets. Rectal swab samples were collected from 83 piglets and weaners in 20 herds and isolated E. coli were characterized by PCR, serotyping and hemolysis. The E. coli strains carried genes for the heat stable toxins STa, STb and EAST1 and adhesins F4 and AIDA-I. The genes for the heat labile toxin LT and adhesins F5, F6, F18 and F41 were not detected in any of the E. coli isolates. Where the serogroup could be identified, E. coli isolates from the same diarrheic pig belonged to the same serogroup. The prevalence of EAST1, STb, Stx2e, STa, AIDA-I, and F4 in the E. coli isolates from suckling piglets and weaners (diarrheic and non-diarrheic combined) was 29, 26.5, 2.4, 1.2, 16, and 8.4 %, respectively. However the prevalence of F4 and AIDA-I in E. coli from diarrheic suckling piglets alone was 22.2 and 20 %, respectively. There was no significant difference in the prevalence of the individual virulence factors in E. coli from the diarrheic and non-diarrheic pigs (p > 0.05). The main ETEC strains isolated from diarrheic and non-diarrheic pigs included F4/STb/EAST1 (7.2 %), F4/STb (1.2 %), AIDA/STb/EAST1 (8 %) and AIDA/STb (8 %). At post-mortem, two diarrheic suckling piglets carrying ETEC showed intact intestinal villi, enterocytes and brush border but with a layer of cells attached to the brush border, suggestive of ETEC infections. This study has shown that the F4 fimbriae is the most predominant in E. coli from diarrheic piglets in the study area and

  3. Ciprofloxacin Treatment Failure in a Murine Model of Pyelonephritis Due to an AAC(6′)-Ib-cr-Producing Escherichia coli Strain Susceptible to Ciprofloxacin In Vitro

    Science.gov (United States)

    Guillard, T.; Cambau, E.; Chau, F.; Massias, L.; de Champs, C.

    2013-01-01

    AAC(6′)-Ib-cr is a plasmid-mediated quinolone resistance mechanism described worldwide for Escherichia coli. Since it confers in vitro only a low level of resistance to ciprofloxacin, we evaluated its impact on the in vivo activity of ciprofloxacin. Isogenic strains were obtained by transferring plasmid p449, harboring aac(6′)-Ib-cr, into the quinolone-susceptible strain E. coli CFT073-RR and its D87G gyrA mutant. MICs were 0.015, 0.06, 0.25, and 0.5 μg/ml against E. coli strains CFT073-RR, CFT073-RR/p449, CFT073-RR GyrAr, and CFT073-RR GyrAr/p449, respectively. Bactericidal activity was reduced at 1× the MIC for the three resistant derivatives, while at a fixed concentration of 0.5 μg/ml, 99.9% killing was observed for all strains except E. coli CFT073-RR GyrAr/p449. In the murine model of pyelonephritis, an optimal regimen of ciprofloxacin (10 mg/kg of body weight twice a day [b.i.d.]) significantly decreased the bacterial count in the kidneys of mice infected with E. coli CFT073 (1.6 versus 4.3 log10 CFU/g of kidney compared to untreated controls; P = 0.0001), while no significant decrease was observed for E. coli CFT073-RR/p449 (2.7 versus 3.1 log10 CFU/g; P = 0.84), E. coli CFT073-RR GyrAr (4.2 versus 4.1 log10 CFU/g; P = 0.35), or E. coli CFT073-RR GyrAr/p449 (2.9 versus 3.6 log10 CFU/g; P = 0.47). While pharmacokinetic and pharmacodynamic (PK/PD) parameters accounted for ciprofloxacin failure against gyrA-containing mutants, this was not the case for the aac(6′)-Ib-cr-containing strains, suggesting an in situ hydrolysis of ciprofloxacin in the latter case. PMID:24018262

  4. Assessment of pit latrines in a peri-urban community in KwaZulu-Natal (South Africa) as a source of antibiotic resistant E. coli strains.

    Science.gov (United States)

    Beukes, Lorika S; King, Tracy L B; Schmidt, Stefan

    2017-08-12

    Due to the frequent use of antibiotics and recurring illnesses related to multidrug-resistant (MDR) bacteria in South Africa, we determined if MDR Escherichia coli were present in pit latrine fecal sludge samples obtained from a peri-urban community in KwaZulu-Natal, South Africa. The abundance of E. coli in pit latrine samples was established using a most probable number (MPN) method with species confirmation done using biochemical tests and polymerase chain reaction (PCR). Forty-four randomly selected E. coli pit latrine isolates were further characterized, using the European committee on antimicrobial susceptibility testing (EUCAST) disk diffusion method to establish antibiotic resistance profiles for these E. coli isolates. The resulting MPN values for E. coli ranged from one to 6.2 log10 MPN per gram of fresh pit latrine fecal sludge. While only 3 out of 44 E. coli pit latrine isolates showed no resistance to any of the 12 tested antibiotics, most isolates were resistant to two or more antibiotics. The majority of isolates showed resistance to at least one of the two tested aminoglycosides, one isolate showed resistance to the carbapenem ertapenem, and although resistance was not detected for tigecycline four pit latrine E. coli isolates showed intermediate resistance to this antibiotic. However, about 14% of the E. coli pit latrine isolates were categorized as MDR, all of which showed resistance to four or more antibiotics. The presence of MDR E. coli strains in pit latrine samples demonstrates that these facilities are potential sources for MDR bacteria. Copyright © 2017 Elsevier GmbH. All rights reserved.

  5. Diarrea neonatal porcina. Aislamiento de cepas de Escherichia coli toxigénicas productoras de STa,