scholarly journals In VitroEffect ofqnrA1,qnrB1, andqnrS1Genes on Fluoroquinolone Activity against IsogenicEscherichia coliIsolates with Mutations ingyrAandparC

2010 ◽  
Vol 55 (3) ◽  
pp. 1266-1269 ◽  
Author(s):  
A. Briales ◽  
J. M. Rodríguez-Martínez ◽  
C. Velasco ◽  
P. Díaz de Alba ◽  
J. Domínguez-Herrera ◽  
...  
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
E Coli ◽  
Mutant Prevention Concentration ◽  
One Step ◽  
Resistant Mutants

ABSTRACTThis article provides an analysis of thein vitroeffect ofqnrA1,qnrB1, andqnrS1genes, combined with quinolone-resistant Ser83Leu substitutions in GyrA and/or Ser80Arg in ParC, on fluoroquinolone (FQ) resistance in isogenicEscherichia colistrains. The association of Ser83Leu substitution in GyrA, Ser80Arg substitution in ParC, andqnrgene expression increased the MIC of ciprofloxacin to 2 μg/ml.qnrgenes present inE. colithat harbored a Ser83Leu substitution in GyrA increased mutant prevention concentration (MPC) values to 8 to 32 μg/ml.qnrgene expression inE. colimay play an important role in selecting for one-step FQ-resistant mutants.

scholarly journals Dual Function of Aar, a Member of the New AraC Negative Regulator Family, in Escherichia coli Gene Expression

2020 ◽  
Vol 88 (6) ◽  
Author(s):  
Abigail S. Mickey ◽  
James P. Nataro
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Negative Regulator ◽  
Dual Function ◽  
Content Type ◽  
Virulence Gene Expression ◽  
E Coli ◽  
K 12

ABSTRACT Enteroaggregative Escherichia coli (EAEC) is an E. coli pathotype associated with diarrhea and growth faltering. EAEC virulence gene expression is controlled by the autoactivated AraC family transcriptional regulator, AggR. AggR activates transcription of a large number of virulence genes, including Aar, which in turn acts as a negative regulator of AggR itself. Aar has also been shown to affect expression of E. coli housekeeping genes, including H-NS, a global regulator that acts at multiple promoters and silences AT-rich genes (such as those in the AggR regulon). Although Aar has been shown to bind both AggR and H-NS in vitro, functional significance of these interactions has not been shown in vivo. In order to dissect this regulatory network, we removed the complex interdependence of aggR and aar by placing the genes under the control of titratable promoters. We measured phenotypic and genotypic changes on downstream genes in EAEC strain 042 and E. coli K-12 strain DH5α, which lacks the AggR regulon. In EAEC, we found that low expression of aar increases aafA fimbrial gene expression via H-NS; however, when aar is more highly expressed, it acts as a negative regulator via AggR. In DH5α, aar affected expression of E. coli genes in some cases via H-NS and in some cases independent of H-NS. Our data support the model that Aar interacts in concert with AggR, H-NS, and possibly other regulators and that these interactions are likely to be functionally significant in vivo.

Characterization of bacteriophages with a lytic effect on various Salmonella serotypes and Escherichia coli O157:H7

2011 ◽  
Vol 57 (12) ◽  
pp. 1042-1051 ◽  
Author(s):  
Osvaldo López-Cuevas ◽  
Nohelia Castro-del Campo ◽  
Josefina León-Félix ◽  
Arturo González-Robles ◽  
Cristóbal Chaidez
Keyword(s):  
Escherichia Coli ◽  
Burst Size ◽  
Growth Curves ◽  
Escherichia Coli O157 ◽  
Genetic Characteristics ◽  
E Coli ◽  
Lytic Effect ◽  
One Step ◽  
Salmonella Serotypes

Four phages isolated from cattle and poultry feces were analyzed for their ability to lyse Salmonella serotypes and Escherichia coli O157:H7. The phage one-step growth curves, morphology, and genetic characteristics were determined. All phages showed a lytic effect on various Salmonella serotypes and E. coli O157:H7, which lysed at least 70% of the 234 strains tested. The phages had latent periods ranging from 10 to 15 min and generation times of 30 to 45 min, while burst size fluctuated between 154 and 426 PFU/cell. Phages morphology showed isometric and elongated heads and rigid contractile tails, consistent with morphology of the Myoviridae family. Phages’ DNA dendrograms showed a distinctive RFLP when digested by HindIII and EcoRV, and SDS–PAGE profile showed distinctive proteins expression as well. In vitro phage challenge showed a total reduction of E. coli O157:H7, Salmonella Typhimurium and Saintpaul counts at 2 h, whereas for Salmonella Montevideo a reduction and retardation growth, at a multiplicity of infection (MOI) of 100, was observed; however, under a MOI of 10 000, no viable cells were detected after 4 h. The wide host ranges of these phages suggested they could be used for simultaneous biocontrol of some Salmonella serotypes and E. coli O157:H7.

scholarly journals Mutant Prevention Concentration Siprofloksasin Terhadap Escherichia coli Patogen dari Kloaka Broiler Secara In Vitro

2021 ◽  
Vol 39 (1) ◽  
pp. 20
Author(s):  
Maria Fatima Palupi ◽  
Eli Nugraha ◽  
Meutia Hayati ◽  
Neneng Atikah
Keyword(s):  
Escherichia Coli ◽  
Bacterial Infections ◽  
Single Step ◽  
In Vitro Test ◽  
Mutant Selection ◽  
E Coli ◽  
Wide Range ◽  
Mutant Prevention Concentration ◽  
Production Animals

Mutant prevention concentration (MPC) is an in vitro test used to determine the lowest drug concentration needed to inhibit the growth of a single-step-mutant bacterial subpopulation. The purpose of this study was to determine the MPC value of ciprofloxacin against pathogenic Escherichia coli to obtained the range of mutant selection windows (MSW) of ciprofloxacin. Ciprofloxacin is a quinolone group that is included in the Highest Priority Critically Important Antimicrobials for Human Medicine but is also used for the treatment of bacterial infections in production animals. Twenty-four of pathogenic E. coli isolates sensitive to ciprofloxacin were tested to obtain MPC values and minimum inhibitory concentration (MIC) values. Test the MPC and MIC values to get the MSW range is done by the method of agar dilution. Mueller-Hinton agar containing standard ciprofloxacin was inoculated with 1010 cfu E. coli for the MPC test and 104 for the MIC test. Based on the MPC test results, the MPC value of ciprofloxacin was 4-64 μg / mL (22.96 ± 19.07 μg / mL) and there was one isolate which had an MPC> 256 μg / mL. These results give a wide range of MSW with a lower limit of the MIC value of 0.25 - 2 µg / mL (0.55 ± 0.37 µg / mL) to the upper limit of the MPC value of 4-64 µg / mL (22.96 ± 19.07 μg / mL). Based on the results of this MPC assessment it can be concluded that the dose of ciprofloxacin in production animals has a wide range of MSW that is allow for single-step mutants.

scholarly journals Efficiency of Single Phage Suspensions and Phage Cocktail in the Inactivation of Escherichia coli and Salmonella Typhimurium: An In Vitro Preliminary Study

2019 ◽  
Vol 7 (4) ◽  
pp. 94 ◽  
Author(s):  
Pedro Costa ◽  
Carla Pereira ◽  
Ana Gomes ◽  
Adelaide Almeida
Keyword(s):  
Escherichia Coli ◽  
Action Spectrum ◽  
E Coli ◽  
Specific Phage ◽  
Salmonella Infections ◽  
Serovar Typhimurium ◽  
Resistant Mutants ◽  
Community Environments ◽  
Enterobacteriaceae Infections

Enterobacteriaceae Escherichia coli and Salmonella enterica serovar Typhimurium strains are among the main pathogens responsible for moderate and serious infections at hospital and community environments, in part because they frequently present resistance to antibiotics. As the treatment of Enterobacteriaceae infections is empiric, using the same antibiotics to treat E. coli and Salmonella infections, the same concept can be applied with phages. The use of different phages combined in cocktails, frequently used to circumvent the development of phage-resistant mutants, also allows for the treatment of multiple pathogens, broadening the phages’ action spectrum. As such, the aim of this study was to evaluate the efficiency of a cocktail of two phages (ELY-1, produced on E. coli and phSE-5, produced on S. Typhimurium) to control E. coli and S. Typhimurium. Phages ELY-1 and phSE-5 were effective against E. coli (maximum reductions of 4.5 and 3.8 log CFU/mL, respectively), S. Typhimurium (maximum reductions of 2.2 and 2.6 log CFU/mL, respectively), and the mixture of both bacteria (maximum reductions of 2.2 and 2.0 log CFU/mL, respectively). The cocktail ELY-1/phSE-5 was more effective against S. Typhimurium and the mixture of both bacteria (maximum reduction of 3.2 log CFU/mL for both) than the single phage suspensions and as effective against E. coli as its specific phage ELY-1 (maximum reductions of 4.5 log CFU/mL). The use of both the phage cocktails, as well as the single-phage suspensions, however, did not prevent the occurrence of phage-resistant mutants. Overall, the results indicate that the application of the phages in the form of a cocktail show their potential to be used presumptively, that is, prior to the identification of the pathogens, paving its use to control E. coli or S. Typhimurium.

scholarly journals The expression of canine cardiac phospholamban in heterologous systems

1989 ◽  
Vol 264 (2) ◽  
pp. 533-538 ◽  
Author(s):  
E A Cook ◽  
J P Huggins ◽  
G Sathe ◽  
P J England ◽  
J R Piggott
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Amino Acids ◽  
Fusion Proteins ◽  
Ns1 Protein ◽  
Native Protein ◽  
E Coli ◽  
Extensive Cell ◽  
Transcription In Vitro

A synthetic phospholamban gene has been cloned and expressed in Escherichia coli, producing both native phospholamban and a fusion protein with 81 amino acids of the influenza virus NS1 protein. Both the native phospholamban and fusion proteins produced extensive cell lysis upon induction of gene expression, but only the native protein underwent spontaneous pentamer formation in E. coli. Translation in vitro of mRNA produced by transcription in vitro of phospholamban cDNA was used to demonstrate the spontaneous aggregation of phospholamban to form pentamers in this system also, both in the presence and absence of exogenous microsomes from canine pancreas or heart. Phospholamban produced by translation in vitro was apparently susceptible to proteolysis by enzymes present in the particulate fractions in these experiments.

scholarly journals Global Gene Expression Profiling of the Asymptomatic Bacteriuria Escherichia coli Strain 83972 in the Human Urinary Tract

2006 ◽  
Vol 74 (6) ◽  
pp. 3565-3575 ◽  
Author(s):  
Viktoria Roos ◽  
Per Klemm
Keyword(s):  
Gene Expression ◽  
Escherichia Coli ◽  
Urinary Tract ◽  
Expression Profiles ◽  
Asymptomatic Bacteriuria ◽  
Global Gene Expression ◽  
E Coli ◽  
Tract Infections

ABSTRACT 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 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 seen between the in vivo expression profiles of strain 83972 in three patients and the corresponding in vitro expression profiles in lab medium and human urine. The data revealed an in vivo lifestyle of microaerobic growth with respiration of nitrate coupled to degradation of sugar acids and amino acids, with no signs of attachment to host tissues. Interestingly, genes involved in NO protection and metabolism showed significant up-regulation in the patients. This is one of the first studies to address bacterial whole-genome expression in humans and the first study to investigate global gene expression of an E. coli strain in the human urinary tract.

scholarly journals Mutant prevention concentrations of ciprofloxacin against urinary isolates of Escherichia coli and Klebsiella pneumoniae

2014 ◽  
Vol 8 (02) ◽  
pp. 154-159 ◽  
Author(s):  
Ziad Daoud ◽  
Elie Salem Sokhn ◽  
Eid Azar ◽  
Khalil Masri ◽  
Shira Doron
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Mueller Hinton ◽  
Mutant Prevention Concentration ◽  
Mueller Hinton Agar ◽  
Resistant Mutants ◽  
Urinary Isolates

Introduction: The aim of this work was to study the ability of ciprofloxacin to restrict the development of resistant mutants of Escherichia coli and Klebsiella pneumoniae through determination of the Mutant Prevention Concentration (MPC). Methodology: We studied 140 strains of E. coli and 86 strains of K. pneumoniae with different profiles of sensitivity to fluoroquinolones and extended spectrum beta lactamase (ESBL) production. The MPCs were determined using an inoculum of 1010 CFU/ml in Mueller-Hinton agar plates with serial concentrations of ciprofloxacin. Results: Ciprofloxacin-susceptible ESBL-producing strains showed a higher MPC for ciprofloxacin (P <0.001) than ciprofloxacin-susceptible non ESBL-producing strains, while ciprofloxacin-resistant ESBL-producing and non ESBL-producing strains did not significantly differ. The presence of qnr variants was associated with elevated MPCs. This was observed for both tested organisms. Conclusions: Our study helps to explain the frequent finding of resistance to fluoroquinolones in ESBL-producing strains. Consequently, the use of concentrations of ciprofloxacin higher than the MIC in order to prevent the recovery and growth of resistant mutants is recommended.

Substituted 4-Formyl-2H-chromen-2-ones: Their Reaction with N-(2,3,4,6-Tetra-Oacetyl- β-D-galactopyranosyl)thiosemicarbazide, Antibacterial and Antifungal Activity of Their Thiosemicarbazone Products

2020 ◽  
Vol 24 (19) ◽  
pp. 2272-2282
Author(s):  
Vu Ngoc Toan ◽  
Nguyen Minh Tri ◽  
Nguyen Dinh Thanh
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Candida Albicans ◽  
Selenium Dioxide ◽  
Antibacterial And Antifungal Activity ◽  
E Coli ◽  
Antibacterial And Antifungal Activities ◽  
Alkyl Ethers ◽  
Antibacterial And Antifungal

Several 6- and 7-alkoxy-2-oxo-2H-chromene-4-carbaldehydes were prepared from corresponding alkyl ethers of 6- and 7-hydroxy-4-methyl-2-oxo-2H-chromen-2-ones by oxidation using selenium dioxide. 6- and 7-Alkoxy-4-methyl-2H-chromenes were obtained with yields of 57-85%. Corresponding 4-carbaldehyde derivatives were prepared with yields of 41-67%. Thiosemicarbazones of these aldehydes with D-galactose moiety were synthesized by reaction of these aldehydes with N-(2,3,4,6-tetra-O-acetyl-β-Dgalactopyranosyl) thiosemicarbazide with yields of 62-74%. These thiosemicarbazones were screened for their antibacterial and antifungal activities in vitro against bacteria, such as Staphylococcus aureus, Escherichia coli, and fungi, such as Aspergillus niger, Candida albicans. Several compounds exhibited strong inhibitory activity with MIC values of 0.78- 1.56 μM, including 8a (against S. aureus, E. coli, and C. albicans), 8d (against E. coli and A. niger), 9a (against S. aureus), and 9c (against S. aureus and C. albicans).

scholarly journals In vitro activity of antimicrobial peptide CDP-B11 alone and in combination with colistin against colistin-resistant and multidrug-resistant Escherichia coli

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Kaitlin S. Witherell ◽  
Jason Price ◽  
Ashok D. Bandaranayake ◽  
James Olson ◽  
Douglas R. Call
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Peptide ◽  
Membrane Integrity ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Resistant Bacteria ◽  
Multidrug Resistant Bacteria ◽  
E Coli ◽  
Minimal Media

AbstractMultidrug-resistant bacteria are a growing global concern, and with increasingly prevalent resistance to last line antibiotics such as colistin, it is imperative that alternative treatment options are identified. Herein we investigated the mechanism of action of a novel antimicrobial peptide (CDP-B11) and its effectiveness against multidrug-resistant bacteria including Escherichia coli #0346, which harbors multiple antibiotic-resistance genes, including mobilized colistin resistance gene (mcr-1). Bacterial membrane potential and membrane integrity assays, measured by flow cytometry, were used to test membrane disruption. Bacterial growth inhibition assays and time to kill assays measured the effectiveness of CDP-B11 alone and in combination with colistin against E. coli #0346 and other bacteria. Hemolysis assays were used to quantify the hemolytic effects of CDP-B11 alone and in combination with colistin. Findings show CDP-B11 disrupts the outer membrane of E. coli #0346. CDP-B11 with colistin inhibits the growth of E. coli #0346 at ≥ 10× lower colistin concentrations compared to colistin alone in Mueller–Hinton media and M9 media. Growth is significantly inhibited in other clinically relevant strains, such as Acinetobacter baumannii, Pseudomonas aeruginosa, and Klebsiella pneumoniae. In rich media and minimal media, the drug combination kills bacteria at a lower colistin concentration (1.25 μg/mL) compared to colistin alone (2.5 μg/mL). In minimal media, the combination is bactericidal with killing accelerated by up to 2 h compared to colistin alone. Importantly, no significant red blood hemolysis is evident for CDP-B11 alone or in combination with colistin. The characteristics of CDP-B11 presented here indicate that it can be used as a potential monotherapy or as combination therapy with colistin for the treatment of multidrug-resistant infections, including colistin-resistant infections.

scholarly journals Lysyl-tRNA synthetase from Escherichia coli K12. Chromatographic heterogeneity and the lysU-gene product

1987 ◽  
Vol 248 (1) ◽  
pp. 43-51 ◽  
Author(s):  
J Charlier ◽  
R Sanchez
Keyword(s):  
Escherichia Coli ◽  
Gene Product ◽  
Activity Ratio ◽  
Deae Cellulose ◽  
Trna Synthetase ◽  
Trna Synthetases ◽  
E Coli ◽  
Aminoacyl Trna Synthetases

In contrast with most aminoacyl-tRNA synthetases, the lysyl-tRNA synthetase of Escherichia coli is coded for by two genes, the normal lysS gene and the inducible lysU gene. During its purification from E. coli K12, lysyl-tRNA synthetase was monitored by its aminoacylation and adenosine(5′)tetraphospho(5′)adenosine (Ap4A) synthesis activities. Ap4A synthesis was measured by a new assay using DEAE-cellulose filters. The heterogeneity of lysyl-tRNA synthetase (LysRS) was revealed on hydroxyapatite; we focused on the first peak, LysRS1, because of its higher Ap4A/lysyl-tRNA activity ratio at that stage. Additional differences between LysRS1 and LysRS2 (major peak on hydroxyapatite) were collected. LysRS1 was eluted from phosphocellulose in the presence of the substrates, whereas LysRS2 was not. Phosphocellulose chromatography was used to show the increase of LysRS1 in cells submitted to heat shock. Also, the Mg2+ optimum in the Ap4A-synthesis reaction is much higher for LysRS1. LysRS1 showed a higher thermostability, which was specifically enhanced by Zn2+. These results in vivo and in vitro strongly suggest that LysRS1 is the heat-inducible lysU-gene product.

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