scholarly journals PREVALENCE AND MOLECULAR CHARACTERIZATION OF blaCTX-M-15-PRODUCING PATHOGENIC GRAM-NEGATIVE BACTERIA FROM VARIOUS CLINICAL SAMPLES

2018 ◽  
Vol 11 (10) ◽  
pp. 386
Author(s):  
Sindhuja S ◽  
Sureshkumar Bt ◽  
Janaki S ◽  
Thenmozhi S
Keyword(s):  
Molecular Characterization ◽  
Pathogenic Bacteria ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Clinical Samples ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
E Coli

Objective: The objective of this study was to describe the prevalence and molecular characterization of blaCTX-M-15-producing pathogenic Gram-negative bacteria from various clinical samples isolated from clinically suspected patients.Methods: In this study, clinical samples of urine, stool, sputum, and pus were collected from 244 patients with nosocomial infections. The phenotypic identification of extended-spectrum β-lactamases (ESBL) was confirmed by double-disk synergy test and combined disk diffusion test. In vitro, the susceptibility pattern of antimicrobial agents against pathogenic isolates was performed by Kirby–Bauer disk diffusion method. The identification of blaCTX-M-15-producing Escherichia coli was assessed by polymerase chain reaction method.Results: The frequency of ESBL-producing pathogenic bacteria from screened was 6 (46.15%). In vitro, susceptibility to pathogenic bacteria showed that the majority of isolates were highly susceptible to amoxicillin-clavulanic acid (97.87%), ofloxacin (93.33%), and Pseudomonas aeruginosa showed 100% sensitive to ceftazidime, cefotaxime, cefixime, cefoperazone, and meropenem (92.30%). The rates of resistance to other antibiotics varied from <26.66%. Among six tested isolates, only one E. coli isolates showed blaCTX-M-15 gene.Conclusion: Due to the increase of E. coli with multiple ESBL genes, continuous surveillance should be needed in clinical field to use of appropriate antibiotics and the control of infections.

scholarly journals Antimicrobial susceptibility of multi-drug and extensively-drug-resistant Escherichia coli to ceftolozane-tazobactam and ceftazidime-avibactam: An in vitro study*

2020 ◽  
Vol 74 ◽  
pp. 77-83
Author(s):  
Patrycja Zalas-Więcek ◽  
Eugenia Gospodarek-Komkowska
Keyword(s):  
Escherichia Coli ◽  
Therapeutic Options ◽  
Clinical Samples ◽  
Gram Negative Bacteria ◽  
Drug Resistant ◽  
In Vitro Susceptibility ◽  
Gram Negative ◽  
E Coli ◽  
Extensively Drug Resistant

Aim: <i>Escherichia coli</i> is one of the Gram-negative bacteria, known to cause many nosocomial infections. Multi-drug (MDR) and extensively-drug resistant (XDR). <i>E. coli</i> are of particular note, due to significant limitations in antibiotic therapy. Ceftolozane-tazobactam and ceftazidime-avibactam are novel therapeutic options against Gram-negative bacteria; hence the aim of this study was to evaluate and compare the <i> in vitro </i> activity of ceftolozane-tazobactam and ceftazidime-avibactam against MDR and XDR clinical <i>E. coli</i> isolates. Material/Methods: The study included 100 non-replicate <i>E. coli</i> isolates derived from clinical samples of patients hospitalized in teaching hospitals. Bacteria were identified by applying mass spectrometry in the MALDI Biotyper system (Bruker). ESBL (bla<sub>CTX-M-1group</sub>, bla<sub>CTX-M-9group</sub>) and carbapenemase (bla<sub>KPC</sub>, bla<sub>VIM</sub>, bla<sub>NDM</sub>, bla<sub>OXA-48</sub>, bla<sub>OXA-181</sub>) genes were detected using the eazyplex® SuperBug CRE test, based on a loop-mediated isothermal amplification (LAMP). The in vitro susceptibility to ceftolozane-tazobactam and ceftazidime-avibactam was tested using validated MIC Test strips (Liofilchem). Results: All 84 extended-spectrum β-lactamase-producing (ESBL) <i>E. coli</i> isolates were susceptible to ceftazidime-avibactam and 83 to ceftolozane-tazobactam. Among 17 <i>E. coli</i> isolates with resistance to at least one of the carbapenems, three (17.6%) were susceptible to ceftolozane-tazobactam and ceftazidime-avibactam. All 14 blaVIM gene-positive <i>E. coli</i> isolates were resistant to both ceftolozane-tazobactam and ceftazidime-avibactam. Both antibiotics were active against bla<sub>CTX-M-9group</sub> and bla<sub>OXA-48</sub> gene-positive <i>E. coli</i> isolates, but they were not active against bla<sub>CTX-M-1group</sub> and bla<sub>VIM</sub> gene-positive isolates. Conclusions: Ceftolozane-tazobactam and ceftazidime-avibactam are alternative, non-carbapenem therapeutic options for ESBL-positive <i>E. coli</i> strains, and they are promising in the treatment of carbapenem-resistant <i>E. coli</i> strains, but not for those carrying the metallo-β-lactamase enzymes. Both drug combinations have comparable activity against ESBL, however, lower MIC values were found for ceftazidime-avibactam.

scholarly journals Antibacterial Effects of Vitamin E: in Vitro Study

2013 ◽  
Vol 7 (2) ◽  
pp. 17-23
Author(s):  
Dalia Abd Al Kader Al-Salih ◽  
Fitua M. Aziz ◽  
Bahir Abdul Razzaq Mshimesh ◽  
Muhanad T. Jehad
Keyword(s):  
Vitamin E ◽  
Pathogenic Bacteria ◽  
In Vitro Study ◽  
The Body ◽  
Diffusion Method ◽  
Gram Negative Bacteria ◽  
Antibacterial Effects ◽  
Gram Positive ◽  
Gram Negative

Overuse of antibiotics has become the major factor for the emergence and dissemination of multi-drug resistant strains of several groups of microorganisms and this lead to search for agents that may have antibacterial effects. Vitamin E emerged as an essential, fat-soluble nutrient in the human body and it is essential, because the body cannot manufacture its own vitamin E, so foods and supplements must provide it. The aim of the present study was to evaluate the effect of vitamin E against pathogenic bacteria. Gram positive and gram negative bacteria were selected as the test microorganisms based on their importance in infections. In this study vitamin E used in four concentrations (50,100,200,400) IU/ml. The agar diffusion method was used to determine antibacterial activity. Results showed that gram negative bacteria were shown to be more resistant than gram positive bacteria. The resistance of gram negative bacteria towards antibacterial substances may be related to lipopolysaccharides in their outer membrane.

scholarly journals 660. Evaluation of Qvella’s FAST-Prep™ Liquid Colony™ for Early Antimicrobial Sensitivity Testing of Positive Blood Culture by Disk Diffusion Method

2020 ◽  
Vol 7 (Supplement_1) ◽  
pp. S386-S386
Author(s):  
Susan M Novak-Weekley ◽  
Aye Aye Khine ◽  
Tino Alavie ◽  
Namidha Fernandez ◽  
Laxman Pandey ◽  
...  
Keyword(s):  
Direct Method ◽  
Viable Cell ◽  
Turnaround Time ◽  
Disk Diffusion ◽  
Diffusion Method ◽  
Gram Negative Bacteria ◽  
Sensitivity Testing ◽  
Gram Positive ◽  
Disk Diffusion Method ◽  
Gram Negative

Abstract Background Conventional antimicrobial susceptibility testing (AST) of microorganisms from positive blood cultures (PBC) can take ≥ 2 days. In order to improve the turnaround time for AST on a PBC, CLSI and EUCAST have made efforts to standardize procedures for disk diffusion (DD) direct from a PBC. Qvella Corporation (Richmond Hill, ON, Canada) has recently developed FAST-Prep, an automated centrifugal sample preparation system that rapidly delivers a Liquid Colony consisting of a purified, concentrated, viable cell suspension directly from a PBC. This study was performed to investigate the feasibility of DD AST off of a PBC using a FAST-Prep Liquid Colony. Methods Contrived PBC samples were prepared by spiking 6 species of Gram-positive and 4 species of Gram-negative bacteria (3-5 strains per species) into FA® Plus bottles and incubating in the BACT/ALERT® VIRTUO® System (bioMerieux, Durham, NC). After positivity, 3 mL of PBC was added to the FAST-Prep cartridge. After 20 minutes of processing in the FAST-Prep instrument, the Liquid Colony was removed from the cartridge and a 0.5 McFarland sample was prepared for DD AST. In parallel, the DD AST from a PBC was performed using 4 drops of PBC (CLSI direct method). Both methods were compared to conventional colony-based DD AST. After 16-18 hours of incubation zone diameters and S/I/R interpretations were determined. Categorical agreement (CA) and errors for both DD AST methods were calculated. In addition, colony plate counting was performed on 0.5 McFarland suspensions of Liquid Colony and the plate colony to determine biomass recovery and sample purity. Results CA for a FAST-Prep DD AST for Gram-positive and Gram-negative bacteria was 95.6% and 98.6%, respectively, compared to CA for CLSI DD AST of 77.2% and 81.9%, respectively. Biomass in the Liquid Colony was 7.2x108 and 1.2x109 CFU for Gram-positive and Gram-negative bacteria, respectively. Cell concentration in the 0.5 McFarland suspension of the Liquid Colony was 3.7x107 and 5.9x107 CFU/mL for Gram-positive and Gram-negative bacteria, respectively, which was similar to the concentration for the reference colony suspension. Conclusion The results support the potential role of FAST-Prep in providing a Liquid Colony for use in rapid AST. Disclosures Susan M. Novak-Weekley, PhD, D(ABMM), Qvella (Employee, Shareholder) Aye Aye Khine, PhD, Qvella (Employee, Shareholder) Tino Alavie, PhD, Qvella (Employee) Namidha Fernandez, MS, Qvella (Employee) Laxman Pandey, MS, Qvella (Employee) Abdossamad Talebpour, PhD, Qvella (Employee, Shareholder)

scholarly journals Interaction of Antimicrobial Peptide Temporin L with Lipopolysaccharide In Vitro and in Experimental Rat Models of Septic Shock Caused by Gram-Negative Bacteria

2006 ◽  
Vol 50 (7) ◽  
pp. 2478-2486 ◽  
Author(s):  
Andrea Giacometti ◽  
Oscar Cirioni ◽  
Roberto Ghiselli ◽  
Federico Mocchegiani ◽  
Fiorenza Orlando ◽  
...  
Keyword(s):  
Septic Shock ◽  
Antimicrobial Peptide ◽  
Gram Negative Bacteria ◽  
Amphibian Skin ◽  
Necrosis Factor Alpha ◽  
Gram Negative ◽  
Rat Models ◽  
E Coli ◽  
Tnf Α

ABSTRACT Sepsis remains a major cause of morbidity and mortality in hospitalized patients, despite intense efforts to improve survival. The primary lead for septic shock results from activation of host effector cells by endotoxin, the lipopolysaccharide (LPS) associated with cell membranes of gram-negative bacteria. For these reasons, the quest for compounds with antiendotoxin properties is actively pursued. We investigated the efficacy of the amphibian skin antimicrobial peptide temporin L in binding Escherichia coli LPS in vitro and counteracting its effects in vivo. Temporin L strongly bound to purified E. coli LPS and lipid A in vitro, as proven by fluorescent displacement assay, and readily penetrated into E. coli LPS monolayers. Furthermore, the killing activity of temporin L against E. coli was progressively inhibited by increasing concentrations of LPS added to the medium, further confirming the peptide's affinity for endotoxin. Antimicrobial assays showed that temporin L interacted synergistically with the clinically used β-lactam antibiotics piperacillin and imipenem. Therefore, we characterized the activity of temporin L when combined with imipenem and piperacillin in the prevention of lethality in two rat models of septic shock, measuring bacterial growth in blood and intra-abdominal fluid, endotoxin and tumor necrosis factor alpha (TNF-α) concentrations in plasma, and lethality. With respect to controls and single-drug treatments, the simultaneous administration of temporin L and β-lactams produced the highest antimicrobial activities and the strongest reduction in plasma endotoxin and TNF-α levels, resulting in the highest survival rates.

Protein expression and extraction of hard-to-produce proteins in the periplasmic space of Escherichia coli v1

2021 ◽  
Author(s):  
Cristina Hernandez Rollan ◽  
Kristoffer Bach Falkenberg ◽  
Maja Rennig ◽  
Andreas Birk Bertelsen ◽  
Morten Norholm
Keyword(s):  
Protein Production ◽  
Expression System ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
E Coli ◽  
New Family ◽  
Lytic Polysaccharide Monooxygenases ◽  
Strain Bl21 ◽  
Polysaccharide Monooxygenases

E. coli is a gram-negative bacteria used mainly in academia and in some industrial scenarios, as a protein production workhorse. This is due to its ease of manipulation and the range of genetic tools available. This protocol describes how to express proteins in the periplasm E. coli with the strain BL21 (DE3) using a T7 expression system. Specifically, it describes a series of steps and tips to express "hard-to-express" proteins in E. coli, as for instance, LPMOs. The protocol is adapted from Hemsworth, G. R., Henrissat, B., Davies, G. J., and Walton, P. H. (2014) Discovery and characterization of a new family of lytic polysaccharide monooxygenases. Nat. Chem. Biol.10, 122–126. .

scholarly journals Antimicrobial Susceptibility Pattern of Gram-Negative Bacterial Isolates from Raw Chicken Meat Samples

2019 ◽  
Vol 6 ◽  
pp. 89-95
Author(s):  
Neha Gautam ◽  
Rojan Poudel ◽  
Binod Lekhak ◽  
Milan Kumar Upreti
Keyword(s):  
Antimicrobial Resistance ◽  
Pathogenic Bacteria ◽  
Chicken Meat ◽  
Food Microbiology ◽  
Diffusion Method ◽  
Resistance Pattern ◽  
Kathmandu Valley ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
Meat Samples

Objectives: This research aims to study the microbial quality of chicken meat available in retail shop of Kathmandu Valley. Methods:  This Study was conducted from June to December 2018 in three different districts of Kathmandu Valley. Samples were collected in sterile plastic bags, labeled properly and stored in an icebox and transported to the Food Microbiology laboratory of Golden Gate International College.  During sample preparation, 25 grams of each sample was taken and transferred to sterile flasks containing 225 ml of buffered peptone water. Potential pathogenic Gram-negative bacteria were isolated by using respective selective media and identified by biochemical test. Antibiotic susceptibility profile of isolates was carried out by Kirby-Bauer disc diffusion method according to CLSI 2017 guideline. Results: Of total 81 chicken meat samples processed; 201 Gram negative bacteria were isolated.  E. coli (100%) was the dominant Gram-negative isolates, followed by Citrobacter spp (62.96%), Pseudomonas spp (40.74%), Proteus spp (19.75%), Salmonella spp (16.04%) and Klebsiella spp (8.64%) respectively. No any multidrug isolates were detected. Conclusion: The study showed that the raw chicken meat samples of Kathmandu valley was highly contaminated with Gram negative potential pathogenic bacteria. Antimicrobial resistance pattern shown by the isolates may indicates that there is not overuse of drug in animals and the less chance of risk of increasing antimicrobial resistance.

scholarly journals 480. Molecular Characterization of Multidrug-Resistant Gram-Negative Pathogens in Three Tertiary Care Hospitals in Egypt

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S235-S235
Author(s):  
Amani Kholy ◽  
Samia A Girgis ◽  
Arwa R Elmanakhly ◽  
Mervat A F Shetta ◽  
Dalia El- Kholy ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Resistance Genes ◽  
Genetic Basis ◽  
Tertiary Care ◽  
Carbapenem Resistance ◽  
Resistance Mechanisms ◽  
Gram Negative ◽  
E Coli ◽  
Tertiary Care Hospitals

Abstract Background High rates of AMR among Gram-negative bacilli (GNB) have been reported from Egypt for almost 2 decades. Surveillance and identifying the genetic basis of AMR provide important information to optimize patient care. As there is no adequate data on the genetic basis of AMR in Egypt, we aimed to identify the molecular characterization of multi-drug-resistant (MDR) Gram-negative pathogens (GNP). Methods Three major tertiary-care hospitals in Egypt participated in the “Study for Monitoring Antimicrobial Resistance Trends” (SMART) from 2014 to 2016. Consecutive GNPs were identified and their susceptibility to antimicrobials were tested. Molecular identification of ESBL, AmpC, and carbapenemase resistance genes was conducted on MDR isolates. Results We enrolled 1,070 consecutive Gram-negative isolates; only one isolate per patient according to the standard protocol of (SMART). During 2014–2015, 578 GNP were studied. Enterobacteriaceae comprised 66% of the total isolates. K. pneumoniae and E. coli were the most common (29.8% and 29.4%). K. pneumoniae and E. coli were the predominant organisms in IAI (30.5% and 30.1%, respectively) and UTI (and 38.9% and 48.6%, respectively), while Acinetobacter baumannii was the most prevalent in RTI (40.2%). ESBL producers were phenotypically detected in 53% of K. pneumoniae, and 68% of E. coli. During 2016, 495 GNP were studied. ESBL continued to be high. For E. coli and K. pneunomiea, the most active antimicrobials were amikacin (≥93%), imipenem/meropenem (≥87%) and colistin (97%). Genetic study of ertapenem-resistant isolates and 50% of isolates with ESBL phenotype revealed ESβL production in more than 90% of isolates; blaCTXM-15 was detected in 71.4% and 68.5% in K. pneumoniae and E. coli, respectively, blaTEM-OSBL in 48.5% and47.5% of K. pneumoniae and E. coli, respectively. Carbapenem resistance genes were detected in 45.4% of isolates. In K. pneumoniae, OXA-48 dominated (40.6%), followed by NDM1 (23.7%) and OXA-232 (4.5%). Conclusion Our study detected alarming rates of resistance and identified many resistance mechanisms in clinical isolates from Egyptian hospitals. These high rates highlight the importance of continuous monitoring of the resistance trend and discovering the novel resistant mechanisms of resistance, and the underscores a national antimicrobial stewardship plan in Egypt. Disclosures All authors: No reported disclosures.

scholarly journals Characterization of the β-lactamase specified by the resistance factor R-1818 in Escherichia coli K12 and other Gram-negative bacteria

1971 ◽  
Vol 123 (4) ◽  
pp. 501-505 ◽  
Author(s):  
J. W. Dale
Keyword(s):  
Escherichia Coli ◽  
Host Species ◽  
Host Bacterium ◽  
Gram Negative Bacteria ◽  
Gram Negative ◽  
E Coli ◽  
R Factor ◽  
Relationship Of ◽  
The Relationship

1. The amino acid composition of the β-lactamase from E. coli (R-1818) was determined. 2. The R-1818 β-lactamase is inhibited by formaldehyde, hydroxylamine, sodium azide, iodoacetamide, iodine and sodium chloride. 3. The Km values for benzylpenicillin, ampicillin and oxacillin have been determined by using the R-factor enzyme from different host species. The same values were obtained, irrespective of the host bacterium. 4. The molecular weight of the enzyme was found to be 44600, and was the same for all host species. 5. The relationship of R-1818 and R-GN238 β-lactamases is discussed.

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