scholarly journals A rapid method to determine the bactericidal activity of compounds against non-replicatingMycobacterium tuberculosisat low pH

2019 ◽  
Author(s):  
Julie V. Early ◽  
Steven Mullen ◽  
Tanya Parish
Keyword(s):  
Bactericidal Activity ◽  
Minimum Bactericidal Concentration ◽  
Treatment Time ◽  
Low Ph ◽  
Citrate Buffer ◽  
Colony Forming Units ◽  
Carbonyl Cyanide ◽  
Viable Bacteria ◽  
Phosphate Citrate Buffer ◽  
Assay Conditions

AbstractThere is an urgent need for new anti-tubercular agents which can lead to a shortened treatment time by targeting persistent or non-replicating bacilli. In order to assess compound activity against non-replicatingMycobacterium tuberculosis, we developed a method to detect the bactericidal activity of novel compounds within 7 days. Our method uses incubation at low pH in order to induce a non-replicating state. We used a strain ofM. tuberculosisexpressing luciferase; we first confirmed the linear relationship between luminescence and viable bacteria (determined by colony forming units) under our assay conditions. We optimized the assay parameters in 96-well plates in order to achieve a reproducible assay. Our final assay usedM. tuberculosisin phosphate-citrate buffer, pH 4.5 exposed to compounds for 7 days; viable bacteria were determined by luminescence. We recorded the minimum bactericidal concentration at pH 4.5 (MBC4.5) representing >2 logs of kill. We confirmed the utility of the assay with control compounds. The ionophores monensin, niclosamide, and carbonyl cyanide 3-chlorophenylhydrazone and the anit-tubercular drugs pretomanid and rifampicin were active, while several other drugs such as isoniazid, ethambutol, and linezolid were not.

scholarly journals Bactericidal Activity, Absence of Serum Effect, and Time-Kill Kinetics of Ceftazidime-Avibactam against β-Lactamase-Producing Enterobacteriaceae and Pseudomonas aeruginosa

2014 ◽  
Vol 58 (9) ◽  
pp. 5297-5305 ◽  
Author(s):  
Tiffany R. Keepers ◽  
Marcela Gomez ◽  
Chris Celeri ◽  
Wright W. Nichols ◽  
Kevin M. Krause
Keyword(s):  
Pseudomonas Aeruginosa ◽  
Human Serum ◽  
Bactericidal Activity ◽  
Minimum Bactericidal Concentration ◽  
Wild Type ◽  
Content Type ◽  
Positive Isolate ◽  
New Treatment ◽  
Time Kill ◽  
Kinetics Of

ABSTRACTAvibactam, a non-β-lactam β-lactamase inhibitor with activity against extended-spectrum β-lactamases (ESBLs), KPC, AmpC, and some OXA enzymes, extends the antibacterial activity of ceftazidime against most ceftazidime-resistant organisms producing these enzymes. In this study, the bactericidal activity of ceftazidime-avibactam against 18Pseudomonas aeruginosaisolates and 15Enterobacteriaceaeisolates, including wild-type isolates and ESBL, KPC, and/or AmpC producers, was evaluated. Ceftazidime-avibactam MICs (0.016 to 32 μg/ml) were lower than those for ceftazidime alone (0.06 to ≥256 μg/ml) against all isolates except for 2P. aeruginosaisolates (1blaVIM-positive isolate and 1blaOXA-23-positive isolate). The minimum bactericidal concentration/MIC ratios of ceftazidime-avibactam were ≤4 for all isolates, indicating bactericidal activity. Human serum and human serum albumin had a minimal effect on ceftazidime-avibactam MICs. Ceftazidime-avibactam time-kill kinetics were evaluated at low MIC multiples and showed time-dependent reductions in the number of CFU/ml from 0 to 6 h for all strains tested. A ≥3-log10decrease in the number of CFU/ml was observed at 6 h for allEnterobacteriaceae, and a 2-log10reduction in the number of CFU/ml was observed at 6 h for 3 of the 6P. aeruginosaisolates. Regrowth was noted at 24 h for some of the isolates tested in time-kill assays. These data demonstrate the potent bactericidal activity of ceftazidime-avibactam and support the continued clinical development of ceftazidime-avibactam as a new treatment option for infections caused byEnterobacteriaceaeandP. aeruginosa, including isolates resistant to ceftazidime by mechanisms dependent on avibactam-sensitive β-lactamases.

scholarly journals THE EFFECT OF PROPOLIS EXTRACT AND PROPOLIS CANDIES ON THE GROWTH OF AGGREGATIBACTER ACTINOMYCETEMCOMITANS ATCC 43718

2017 ◽  
Vol 10 (17) ◽  
pp. 26
Author(s):  
Khodijah Khodijah ◽  
Ratna Farida ◽  
Nurtami Soedarsono
Keyword(s):  
Minimum Inhibitory Concentration ◽  
Inhibitory Concentration ◽  
Minimum Bactericidal Concentration ◽  
Aggregatibacter Actinomycetemcomitans ◽  
Spectrophotometric Analysis ◽  
Propolis Extract ◽  
Post Exposure ◽  
Colony Forming Units

Objective: This experiment aimed to analyze the effect of propolis extract and propolis containing candies on the growth of Aggregatibacter actinomycetemcomitans using spectrophotometric analysis and colony-forming units (CFU) counts.Methods: After A. actinomycetemcomitans were exposed to propolis extract and candies, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) were determined with spectrophotometry and post-exposure colony counting.Results: The MIC of propolis extract against A. actinomycetemcomitans was determined to be 10%, and the MBC was 20%. A decrease in the total CFU count of A. actinomycetemcomitans was observed after propolis extract and candy exposure.Conclusions: Propolis extract and propolis candies were effective in inhibiting the growth of A. actinomycetemcomitans ATCC 43718 in vitro.

scholarly journals The Bactericidal Activity and Spore Inhibition Effect of Manuka Honey against Clostridioides Difficile

Antibiotics ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 684
Author(s):  
Lillian Yu ◽  
Reynal Palafox-Rosas ◽  
Brian Luna ◽  
Rosemary C. She
Keyword(s):  
Bactericidal Activity ◽  
Viable Cell ◽  
Manuka Honey ◽  
Inhibition Effect ◽  
Natural Substances ◽  
Clostridioides Difficile ◽  
Cell Counts ◽  
Colony Forming Units ◽  
Conventional Antibiotics ◽  
Drug Free

Clostridioides difficile colitis overgrowth occurs when the normal gut microbiome becomes disrupted, often due to antibiotics. Effective treatment remains elusive, due partly to the persistence of its spores in the gut. Natural substances like manuka honey offer an alternative antimicrobial mechanism of action to conventional antibiotics. We investigated the antibiotic activity of manuka honey against 20 C. difficile isolates. The minimum inhibitory concentrations (MICs) and minimal bactericidal concentrations (MBC) of manuka honeys of methylglyoxal (MGO) grades 30+, 100+, 250+, and 400+ were determined based on broth microdilution. Sporicidal activity was assessed in a range of honey concentrations by enumerating total viable cell and spore counts at 0–96 h after organism inoculation. The MICs of C. difficile ranged from 4% to >30% (w/v). MIC50 for the four MGO grades were similar at 10–14%. MBC results for the majority of isolates were distributed bimodally at MBC/MIC ratios ≤4 or MBC >30%. Growth kinetics in honey showed total viable cell counts remaining >105 colony-forming units (CFU)/mL at all time points, whereas spore counts remained within 1-log of baseline (102 CFU/mL) in honey but steadily increased in the drug-free control to >105 CFU/mL by 96 h. Manuka honey demonstrated variable inhibitory and bactericidal activity against C. difficile. MGO grade had no noticeable impact on overall MIC distributions or bactericidal activity. Although manuka honey could inhibit spore proliferation, it did not eradicate spores completely.

Preparation of a Factor VII Concentrate

1979 ◽  
Author(s):  
A.J. MacLeod ◽  
I. Dickson
Keyword(s):  
Specific Activity ◽  
Fresh Frozen Plasma ◽  
Factor Vii ◽  
Batch Adsorption ◽  
Citrate Buffer ◽  
Linear Gradient ◽  
Fresh Frozen ◽  
Frozen Plasma ◽  
Phosphate Citrate Buffer

A factor VII concentrate has been prepared from pooled citrated fresh frozen plasma following removal of cryoprecipitate and factors II, IX and X. The method involved batch adsorption on DEAE-Sephadex A-50, fractionation of the subsequent batch eluate by PEG precipitation and passage through a column of DEAE-Sepharose CL-.6B. A phosphate-citrate buffer pH 6.9 was used throughout, this was made 0.2M with NaCl for the batch elution and a 0 - 0.2H NaCl linear gradient was used to elute the components from the column. Factor VII activity was clearly resolved from the bulk of the protein, including caeruloplasmin, and could be recovered as a concentrate at about 20 U FVII/ml with a specific activity of in excess of 1 U FVII/mg of protein and an overall recovery of 40% to 50%

Concurrent Effects of High Hydrostatic Pressure, Acidity and Heat on the Destruction and Injury of Yeasts

1995 ◽  
Vol 58 (3) ◽  
pp. 301-304 ◽  
Author(s):  
YOGA PANDYA ◽  
FRED F. JEWETT ◽  
DALLAS G. HOOVER
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Hydrostatic Pressure ◽  
High Hydrostatic Pressure ◽  
Potato Dextrose Agar ◽  
Plate Count ◽  
Citrate Buffer ◽  
Zygosaccharomyces Bailii ◽  
Mild Heat ◽  
Yeast Cultures ◽  
Colony Forming Units

Saccharomyces cerevisiae ATCC 2373 and Zygosaccharomyces bailii ATCC 36947 were exposed to hydrostatic pressures ranging from 1,500 to 3,000 atmospheres for 10, 20 and 30 min in 0.1 M citrate buffer at pH 3.0, 4.0 and 5.0 at 25 and 45°C. Inactivation of inoculated yeast cultures was achieved in spaghetti sauce with meat at 25°C with 3,000 atmospheres for 10 min and also at 45°C and 2,500 atmospheres for 10 min. Viable counts were determined on potato dextrose agar (PDA) incubated at 30°C for 48 h. Pressure-induced injury was demonstrated by plate count differential between PDA and PDA supplemented with glucose (PDAG). A reduction of 7-log10 cycles colony forming units (CFU)/ml was seen for both strains at 3,000 atmospheres for 10 min at 25°C at all pH levels and at 2,250 atmospheres, pH 5.0 for 20 min at 45°C. At 2,000 atmospheres, pH 3.0 for 30 min, the increase in temperature from 25 to 45°C increased the inactivation of yeast by 6-log10 cycles. Lowering the pH from 5.0 to 3.0 enhanced lethality up to 2-log10 cycles at 2,250 atmospheres, 25°C for 30 min. Injury was most apparent at exposure parameters that produced 3- to 5-log10 cycle reductions on PDA. This was achieved (99% injury) at 2,250 atmospheres, 25°C for 30 min. These data indicate that mild heat and acidity contribute to the effectiveness of the inactivation and injury of yeast by high hydrostatic pressure (HHP).

scholarly journals Evaluation of the Bactericidal Activity of Plazomicin and Comparators against Multidrug-Resistant Enterobacteriaceae

2018 ◽  
Vol 62 (8) ◽  
Author(s):  
M. Thwaites ◽  
D. Hall ◽  
D. Shinabarger ◽  
A. W. Serio ◽  
K. M. Krause ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Klebsiella Pneumoniae ◽  
Bactericidal Activity ◽  
Minimum Bactericidal Concentration ◽  
Multidrug Resistant ◽  
Resistance Mechanisms ◽  
Next Generation ◽  
Content Type ◽  
Time Kill

ABSTRACT The next-generation aminoglycoside plazomicin, in development for infections due to multidrug-resistant (MDR) Enterobacteriaceae, was evaluated alongside comparators for bactericidal activity in minimum bactericidal concentration (MBC) and time-kill (TK) assays against MDR Enterobacteriaceae isolates with characterized aminoglycoside and β-lactam resistance mechanisms. Overall, plazomicin and colistin were the most potent, with plazomicin demonstrating an MBC50/90 of 0.5/4 μg/ml and sustained 3-log10 kill against MDR Escherichia coli, Klebsiella pneumoniae, and Enterobacter spp.

Determination of fumonisins B1 and B2 in maize-based baby food products by HPLC with fluorimetric detection after immunoaffinity column clean-up

2010 ◽  
Vol 3 (2) ◽  
pp. 135-146 ◽  
Author(s):  
A. De Girolamo ◽  
D. Pereboom-de Fauw ◽  
E. Sizoo ◽  
H. van Egmond ◽  
L. Gambacorta ◽  
...  
Keyword(s):  
High Performance ◽  
Signal To Noise Ratio ◽  
Retail Market ◽  
Immunoaffinity Column ◽  
Limit Of Quantification ◽  
Citrate Buffer ◽  
Relative Standard ◽  
Phosphate Citrate Buffer ◽  
Infants And Young Children

A method for the determination of fumonisin B1 (FB1) and B2 (FB2) in different commercial maize-based products for infants and young children was developed and tested in a limited validation study involving 3 laboratories. The method used extraction at 55 °C with an acidic mixture of methanol-acetonitrile-phosphate/citrate buffer, clean-up through immunoaffinity column and fumonisin determination by high performance liquid chromatography with automated pre-column derivatisation with o-phthaldialdehyde. Recovery experiments were performed at five spiking levels in the ranges of 80-800 µg/kg FB1 and 20-200 µg/kg FB2. Mean recoveries ranged from 83 to 97% for FB1 and from 61 to 78% for FB2. Relative standard deviations for within-laboratory repeatability (RSDr) ranged from 5 to 12% for FB1 and from 8 to 13% for FB2, whereas relative standard deviation for between-laboratory reproducibility (RSDR) ranged from 6 to 10% for FB1 and from 9 to 16% for FB2. The limit of quantification of the method (signal to noise ratio of 6) was 2.8 µg/kg for FB1 and 2.2 µg/kg for FB2. Fumonisins were found in 6 out of 19 maize-based baby foods obtained from the Italian retail market at levels up to 53 µg/kg.

Serum Inhibitory and Bactericidal Activity of Ciprofloxacin following Intravenous Administration

DICP ◽  
1989 ◽  
Vol 23 (6) ◽  
pp. 456-460
Author(s):  
Michael N. Dudley ◽  
Hilary D. Mandler ◽  
Kenneth H. Mayer ◽  
Stephen H. Zinner
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Pseudomonas Aeruginosa ◽  
Minimum Inhibitory Concentration ◽  
Bactericidal Activity ◽  
Resistant Strain ◽  
Inhibitory Concentration ◽  
Minimum Bactericidal Concentration ◽  
E Coli ◽  
Dose Levels

Serum inhibitory and bactericidal titers were measured in nine healthy volunteers following single iv doses of ciprofloxacin 100, 150, and 200 mg. The median peak serum bactericidal titer (5 minutes following completion of a 30-minute infusion) against two highly susceptible strains of Escherichia coli ranged between 1:64 and 1:1024 and titers exceeded 1:8 for six hours for all dose levels. The bactericidal titers against two strains of Pseudomonas aeruginosa and a methicillin-resistant strain of Staphylococcus aureus were considerably lower, the median peak being 1:2 at all dose levels. Measured inhibitory and bactericidal titers at five minutes and one hour postinfusion were significantly greater than those predicted (measured serum ciprofloxacin concentration to minimum inhibitory concentration [MIC] or minimum bactericidal concentration [MBC]) for only one strain of E. coli. Intravenous doses of ciprofloxacin 100–200 mg produce high and sustained serum bactericidal titers against highly susceptible bacteria; considerably lower levels of activity are seen against bacteria having higher MICs and MBCs but still considered susceptible to the drug.

Anti-Escherichia coliO157:H7 activity of free fatty acids under varying pH

2010 ◽  
Vol 56 (3) ◽  
pp. 263-267 ◽  
Author(s):  
Jinli Yang ◽  
Xianzhi Hou ◽  
Priya S. Mir ◽  
Tim A. McAllister
Keyword(s):  
Fatty Acids ◽  
Bactericidal Activity ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Capric Acid ◽  
Luria Bertani ◽  
E Coli ◽  
Luria Bertani Broth ◽  
Colony Forming Units ◽  
Acid Tolerant

Following screening of 4 strains of Escherichia coli O157:H7 (E32511, E318N, H4420N, and R508N) for acid tolerance, strain H4420N was selected for further study into the influence of pH on bactericidal activity of 6 fatty acids (capric, lauric, palmitic, oleic, linoleic, and linolenic). Strain H4420N was cultured for 6 h in Luria–Bertani broth amended with individual fatty acids at 20 mmol/L, with pH adjusted to 7.0, 4.3, or 2.5. None of the fatty acids exhibited bactericidal activity at pH 7.0 (p >0.05). At pH 4.3, only capric, lauric, and linoleic acids reduced viability of H4420N (p < 0.05). At pH 2.5, oleic (C18:1) and linolenic (C18:3) acids had modest effects on H4420N viability, whereas capric (C10:0), lauric (C12:0), and linoleic (C18:2) acids resulted in a reduction ≥5 log10colony-forming units (CFU)/mL (p < 0.05). Capric and lauric acids were examined further at pH 2.5 over a range of concentrations (0.15–20 mmol/L). After 10 min of exposure, 5 log10 CFU/mL reductions (p < 0.05) were achieved by lauric acid at 2.5 mmol/L and by capric acid at 0.31 mmol/L. Acid stress increased the sensitivity of acid-tolerant E. coli O157:H7 strain H4420N to fatty acids. Including sources of these fatty acids in diets for cattle might impair the ability of this zoonotic pathogen to survive passage through the stomach, possibly reducing the potential for its colonization in the lower gut.

scholarly journals Evaluation of the disinfecting capacity of ozone in emergency vehicles

2020 ◽  
Author(s):  
Jorge Biurrun Cía ◽  
Begoña García Martínez ◽  
Andrea Perez Montero ◽  
Grazyna Kochan ◽  
David Escors ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Lentiviral Vector ◽  
Ozone Treatment ◽  
Bacterial Strains ◽  
Health Crisis ◽  
Colony Forming Units ◽  
Objective Evidence ◽  
Emergency Vehicles ◽  
Viable Bacteria ◽  
Pre Treatment

ABSTRACTObjectiveAs a consequence of the health crisis arising from the SARS-CoV-2 coronavirus pandemic, ozone treatments are being applied as disinfectant in emergency vehicles, without objective evidence on its efficacy. Here we evaluate the efficacy of ozone treatment over bacterial strains and virus-like particles.MethodA preparation of a lentiviral vector (lentivector) and dried cultures of two bacterial strains (gram + Staphylococcus aureus and gram - Salmonella enterica ser. Enteritidis) were placed inside an ambulance at two different locations. The interior of the vehicle was subjected to 10 min and 20 min treatments (3 and 6 times the recommended time by the manufacturer). Following the treatments, lentivector preparations were titrated, and viable bacteria (colony forming units, CFUs) counted and compared to pre-treatment titers and infectious CFUs of the same lysates and cultures.ResultsNone of the treatments significantly reduced either lentivector titer or the number of viable bacteria.ConclusionsAt least in the analyzed conditions and for the microorganisms used in this study, it can be concluded that ozone treatment is not advisable for the disinfection of emergency vehicles.

Sign in / Sign up

Export Citation Format

Share Document