scholarly journals Rapid Detection of Escherichia coli Antibiotic Susceptibility Using Live/Dead Spectrometry for Lytic Agents

2021 ◽  
Vol 9 (5) ◽  
pp. 924
Author(s):  
Julia Robertson ◽  
Cushla McGoverin ◽  
Joni R. White ◽  
Frédérique Vanholsbeeck ◽  
Simon Swift
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Rapid Detection ◽  
Inhibitory Action ◽  
Detection Method ◽  
Treatment Time ◽  
Polymyxin B ◽  
Diagnostic Techniques ◽  
Antibiotic Concentration ◽  
Syto 9

Antibiotic resistance is a serious threat to public health. The empiric use of the wrong antibiotic occurs due to urgency in treatment combined with slow, culture-based diagnostic techniques. Inappropriate antibiotic choice can promote the development of antibiotic resistance. We investigated live/dead spectrometry using a fluorimeter (Optrode) as a rapid alternative to culture-based techniques through application of the LIVE/DEAD® BacLightTM Bacterial Viability Kit. Killing was detected by the Optrode in near real-time when Escherichia coli was treated with lytic antibiotics—ampicillin and polymyxin B—and stained with SYTO 9 and/or propidium iodide. Antibiotic concentration, bacterial growth phase, and treatment time used affected the efficacy of this detection method. Quantification methods of the lethal action and inhibitory action of the non-lytic antibiotics, ciprofloxacin and chloramphenicol, respectively, remain to be elucidated.

scholarly journals Antibiotic breakdown by susceptible bacteria enhances the establishment of β-lactam resistant mutants

2021 ◽  
Author(s):  
Manja Saebelfeld ◽  
Suman G Das ◽  
Jorn Brink ◽  
Arno Hagenbeek ◽  
Joachim Krug ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Theoretical Model ◽  
Enzymatic Activity ◽  
Social Interactions ◽  
Single Cells ◽  
Antibiotic Concentration ◽  
High Frequencies ◽  
Establishment Probability ◽  
Resistant Mutants

For a better understanding of the evolution of antibiotic resistance, it is imperative to study the factors that determine the initial establishment of mutant resistance alleles. In addition to the antibiotic concentration, the establishment of resistance alleles may be affected by interactions with the surrounding susceptible cells from which they derive, for instance via the release of nutrients or removal of the antibiotic. Here, we investigate the effects of social interactions with surrounding susceptible cells on the establishment of Escherichia coli mutants with increasing β-lactamase activity (i.e. the capacity to hydrolyze β-lactam antibiotics) from single cells under the exposure of the antibiotic cefotaxime on agar plates. We find that mutant establishment probability is increased in the presence of susceptible cells due to the active breakdown of the antibiotic, but the rate of breakdown by the susceptible strain is much higher than expected based on its low enzymatic activity. A detailed theoretical model suggests that this observation can be explained by cell filamentation causing delayed lysis. While susceptible cells may hamper the spread of higher-resistant β-lactamase mutants at relatively high frequencies, our findings show that they could promote establishment during their emergence.

scholarly journals Rapid Detection Method for Enteroaggregative <i>Escherichia coli</i> Using Simple Clump Formation and Aggregative Assay

2013 ◽  
Vol 03 (08) ◽  
pp. 552-556
Author(s):  
Miyuki Fujioka ◽  
Chowdhury Rafiqul Ahsan ◽  
Yoshimitsu Otomo
Keyword(s):  
Escherichia Coli ◽  
Rapid Detection ◽  
Detection Method

scholarly journals Antibiotic Breakdown by Susceptible Bacteria Enhances the Establishment of β-Lactam Resistant Mutants

2021 ◽  
Vol 12 ◽  
Author(s):  
Manja Saebelfeld ◽  
Suman G. Das ◽  
Jorn Brink ◽  
Arno Hagenbeek ◽  
Joachim Krug ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Theoretical Model ◽  
Social Interactions ◽  
Single Cells ◽  
Antibiotic Concentration ◽  
Activity Increase ◽  
High Frequencies ◽  
Resistant Mutants ◽  
Mutant Cells

For a better understanding of the evolution of antibiotic resistance, it is imperative to study the factors that determine the initial establishment of mutant resistance alleles. In addition to the antibiotic concentration, the establishment of resistance alleles may be affected by interactions with the surrounding susceptible cells from which they derive, for instance via the release of nutrients or removal of the antibiotic. Here, we investigate the effects of social interactions with surrounding susceptible cells on the establishment of Escherichia coli mutants with increasing β-lactamase activity (i.e., the capacity to hydrolyze β-lactam antibiotics) from single cells under the exposure of the antibiotic cefotaxime (CTX) on agar plates. We find that relatively susceptible cells, expressing a β-lactamase with very low antibiotic-hydrolyzing activity, increase the probability of mutant cells to survive and outgrow into colonies due to the active breakdown of the antibiotic. However, the rate of breakdown by the susceptible strain is much higher than expected based on its low enzymatic activity. A detailed theoretical model suggests that this observation may be explained by cell filamentation causing delayed lysis. While susceptible cells may hamper the spread of higher-resistant β-lactamase mutants at relatively high frequencies, our findings show that they promote their initial establishment.

Environmental pollutants induce noninherited antibiotic resistance to polymyxin B in Escherichia coli

2020 ◽  
Vol 15 (17) ◽  
pp. 1631-1643
Author(s):  
Dorin Harpaz ◽  
Robert S Marks ◽  
Ariel Kushmaro ◽  
Evgeni Eltzov
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Bacterial Resistance ◽  
Environmental Pollutants ◽  
Polymyxin B ◽  
Environmental Toxicants ◽  
Toxic Compounds ◽  
Resistance To Antibiotics ◽  
Low Concentrations ◽  
The Impact

Aim: The mechanisms behind antibiotic resistance by bacteria are important to create alternative molecules. Objective: This study focuses on the impact of environmental pollutants on bacterial resistance to antibiotics. Materials & methods: The effect of various environmental pollutants on noninherited bacterial resistance to antibiotics was examined. Results: The tolerance to the polymyxin-B antibiotic was shown to be conferred to Escherichia coli, by pretreatment with subinhibitory concentrations of environmental toxicants. The cell survival to a sublethal dosage of antibiotics was tested. Exposure to low concentrations of toxic compounds (500 ppb copper, 2% [v/v] ethanol or 0.5 μg/ml trimethoprim) stimulated the bacterial heat shock systems and led to increased tolerance to polymyxin B. Conclusion: Environmental pollutants induce a temporary bacterial noninheritable resistance to antibiotic.

Antibiotic-resistant Bacteria in Raw Milk and Ability of Some to Transfer Antibiotic Resistance to Escherichia coli

1979 ◽  
Vol 42 (12) ◽  
pp. 950-953 ◽  
Author(s):  
LESTER HANKIN ◽  
GEORGE H. LACY ◽  
GEORGE R. STEPHENS ◽  
WALTER F. DILLMAN
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Raw Milk ◽  
Significant Negative Correlation ◽  
Polymyxin B ◽  
Resistant Bacteria ◽  
Antibiotic Resistant Bacteria ◽  
Antibiotic Resistant ◽  
Milk Samples ◽  
Neomycin Sulfate

Raw milk samples were examined for number and percentage of bacteria resistant to seven antibiotics: penicillin, ampicillin, chloramphenicol, neomycin sulfate, polymyxin B sulfate, tetracycline and streptomycin sulfate. A significant negative correlation was found between the total aerobic count of the milk sample and the concentration (above 5 or 10% of the total count) of bacteria in each milk resistant to each of the antibiotics tested. Three of 42 gram-negative isolates were capable of transferring their antibiotic resistance to Escherichia coli. Substantial numbers of antibiotic-resistant bacteria in raw milk were found and some survived pasteurization. Inspection of farms failed to indicate a relationship between farm practices or use of antibiotics in feed or as pharmaceuticals and number of antibiotic-resistant bacteria in the raw milk.

scholarly journals Development of Molecular Rapid Detection for Vibrio cholerae and Escherichia coli

2020 ◽  
Author(s):  
Diana Elizabeth Waturangi ◽  
JASON PETRUS ◽  
RICO KOSASIH ◽  
GLORIA RAISSA
Keyword(s):  
Escherichia Coli ◽  
Vibrio Cholerae ◽  
False Positive ◽  
Rapid Detection ◽  
Pathogenic Bacteria ◽  
Molecular Detection ◽  
Detection Method ◽  
False Negative ◽  
E Coli ◽  
Drinking Water Samples

Abstract Background: Vibrio cholerae and Escherichia coli were main causative agent foodborne diseases, especially in many developing countries, such as Indonesia. Thereby, rapid detection of these pathogenic bacteria is necessary to quickly detect infection that occurred so it can be treated immediately. In this case, multiplex PCR allows multiple genes amplification in one reaction thereby enable to perform rapid detection of these pathogenic bacteria. The objective of this study is to develop rapid molecular detection of V. Cholerae and E. coli and analyze the sensitivity and specificity of this assay.Result: In this study, we used various virulence genes in each pathogenic bacteria as marker to develop rapid molecular detection. Based on this research, optimum results of V. cholerae and E. coli rapid detection were obtained with a primer concentration of 16 µM for ctxA and ompU, 30 µM for ace, and 50 µM for zot, and toxR; 2 µM for elt and 5 µM for stx, respectively. Finally, based on the method standardization by ISO/TS 20836 these assays had 0% false positive, 0% false negative, 100% specificity, and 100% sensitivity; 0% false positive, 4% false negative, 100% specificity, and 96% sensitivity for V. cholerae and E. coli respectively. Conclusion: The optimized method was qualified to be used as a detection method for V. cholerae and E. coli detection according to ISO/TS 20836 (2017) and EHEC and ETEC contamination in drinking water samples.

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