Influence of Temperature, Glucose, and Iron on Sinigrin Degradation by Salmonella and Listeria monocytogenes

2014 ◽  
Vol 77 (12) ◽  
pp. 2133-2138 ◽  
Author(s):  
AMIN N. OLAIMAT ◽  
BABAK SOBHI ◽  
RICHARD A. HOLLEY
Keyword(s):  
Listeria Monocytogenes ◽  
Ferrous Iron ◽  
Ferric Iron ◽  
Allyl Isothiocyanate ◽  
Effect Of Temperature ◽  
Iron Compounds ◽  
Mueller Hinton ◽  
New Compounds ◽  
Hydrolysis Of ◽  
Mueller Hinton Broth

Factors, including pH, temperature, glucose concentration, and iron compounds, affect the activity of plant myrosinase and, consequently, endogenous glucosinolate degradation. These factors also may affect glucosinolate degradation by bacterial myrosinase. Therefore, this study examined the effect of temperature (4 to 21°C), glucose (0.05 to 1.0%), and iron (10 mM ferrous or ferric) on sinigrin degradation by Salmonella or Listeria monocytogenes cocktails in Mueller-Hinton broth and the effect of sinigrin degradation on bacterial viability. The degradation of sinigrin by both pathogens increased with higher temperatures (21 > 10 > 4°C). Salmonella and L. monocytogenes cocktails hydrolyzed 59.1 and 53.2% of sinigrin, respectively, at 21°C up to 21 days. Both iron compounds significantly enhanced sinigrin degradation by the pathogens. On day 7, sinigrin was not detected when the Salmonella cocktail was cultured with ferrous iron or when the L. monocytogenes cocktail was cultured in Mueller-Hinton broth containing ferric iron. In contrast, ferric and ferrous iron inhibited the activity of 0.002 U/ml myrosinase from white mustard by 63 and 35%, respectively, on day 1. Salmonella and L. monocytogenes cocktails were able to degrade >80% of sinigrin at 0.05 and 0.1% glucose; however, 0.25 to 1.0% glucose significantly reduced sinigrin degradation. Although both pathogens significantly degraded sinigrin, the allyl isothiocyanate (AITC) recoverable was ≤6.2 ppm, which is not inhibitory to Salmonella or L. monocytogenes. It is probable that the gradual hydrolysis of sinigrin to form AITC either did not produce an inhibitory level of AITC or the AITC formed was unstable in the aqueous medium and rapidly decomposed to new compounds that were less bactericidal against the pathogens.

Reduction of ferric iron by Listeria monocytogenes and other species of Listeria

1993 ◽  
Vol 39 (5) ◽  
pp. 480-485 ◽  
Author(s):  
Harry G. Deneer ◽  
Irene Boychuk
Keyword(s):  
Listeria Monocytogenes ◽  
Ferrous Iron ◽  
Ferric Iron ◽  
Agar Plate ◽  
Ammonium Citrate ◽  
Ferric Ammonium Citrate ◽  
Flavin Mononucleotide ◽  
Ferric Reductase ◽  
Iron Source ◽  
Plate Assay

One mechanism by which Listeria monocytogenes is thought to obtain iron required for growth is through the extracellular reduction of a ferric iron source to the ferrous form. To better characterize this reductase activity we have developed a simple plate assay that allows detection of colonies of Listeria species able to reduce ferric iron. Cells are plated on an agar base medium containing a ferric iron source and ethylenediamine dihydroxyphenylacetic acid. Colonies are then overlain with soft agarose containing NADH, flavin mononucleotide, and Ferrozine, a chelator of ferrous iron. Colonies able to reduce the ferric iron source form a red-purple color as the ferrous iron is complexed with ferrozine. Using this qualitative assay we have shown that all species of Listeria are able to reduce ferric iron when presented as ferric ammonium citrate whereas most other species of Gram-positive and Gram-negative bacteria are not. Only Clostridium perfringens was able to reduce ferric iron to the same extent as Listeria. Listeria monocytogenes was further shown to be capable of reducing various ferric iron salts as well as iron bound to ferritin, transferrin, and 2,3-dihydroxybenzoic acid in the agar plate assay. The utility of this assay was demonstrated by using it to screen a bank of Tn9/6-derived mutants of L. monocytogenes for clones unable to reduce ferric iron. Four such mutants were identified and all were shown to have greatly decreased ferric reductase activity.Key words: Listeria, ferric reductase, reductase mutants.

scholarly journals Recording the Presence of Peanibacillus larvae larvae Colonies on MYPGP Substrates Using a Multi-Sensor Array Based on Solid-State Gas Sensors

Sensors ◽  
2021 ◽  
Vol 21 (14) ◽  
pp. 4917
Author(s):  
Beata Bąk ◽  
Jakub Wilk ◽  
Piotr Artiemjew ◽  
Jerzy Wilde
Keyword(s):  
Gas Sensors ◽  
Culture Media ◽  
Laboratory Diagnosis ◽  
Baseline Correction ◽  
American Foulbrood ◽  
Sensor Signal ◽  
Sodium Pyruvate ◽  
Mueller Hinton ◽  
Regeneration Phase ◽  
Mueller Hinton Broth

American foulbrood is a dangerous disease of bee broods found worldwide, caused by the Paenibacillus larvae larvae L. bacterium. In an experiment, the possibility of detecting colonies of this bacterium on MYPGP substrates (which contains yeast extract, Mueller-Hinton broth, glucose, K2HPO4, sodium pyruvate, and agar) was tested using a prototype of a multi-sensor recorder of the MCA-8 sensor signal with a matrix of six semiconductors: TGS 823, TGS 826, TGS 832, TGS 2600, TGS 2602, and TGS 2603 from Figaro. Two twin prototypes of the MCA-8 measurement device, M1 and M2, were used in the study. Each prototype was attached to two laboratory test chambers: a wooden one and a polystyrene one. For the experiment, the strain used was P. l. larvae ATCC 9545, ERIC I. On MYPGP medium, often used for laboratory diagnosis of American foulbrood, this bacterium produces small, transparent, smooth, and shiny colonies. Gas samples from over culture media of one- and two-day-old foulbrood P. l. larvae (with no colonies visible to the naked eye) and from over culture media older than 2 days (with visible bacterial colonies) were examined. In addition, the air from empty chambers was tested. The measurement time was 20 min, including a 10-min testing exposure phase and a 10-min sensor regeneration phase. The results were analyzed in two variants: without baseline correction and with baseline correction. We tested 14 classifiers and found that a prototype of a multi-sensor recorder of the MCA-8 sensor signal was capable of detecting colonies of P. l. larvae on MYPGP substrate with a 97% efficiency and could distinguish between MYPGP substrates with 1–2 days of culture, and substrates with older cultures. The efficacy of copies of the prototypes M1 and M2 was shown to differ slightly. The weighted method with Canberra metrics (Canberra.811) and kNN with Canberra and Manhattan metrics (Canberra. 1nn and manhattan.1nn) proved to be the most effective classifiers.

scholarly journals In Vitro Activities of Daptomycin against 2,789 Clinical Isolates from 11 North American Medical Centers

2001 ◽  
Vol 45 (6) ◽  
pp. 1919-1922 ◽  
Author(s):  
Arthur L. Barry ◽  
Peter C. Fuchs ◽  
Steven D. Brown
Keyword(s):  
Clinical Isolates ◽  
In Vitro Activity ◽  
Broth Microdilution ◽  
In Vitro Susceptibility ◽  
Medical Centers ◽  
Mueller Hinton ◽  
Calcium Cations ◽  
Important Exception ◽  
Mueller Hinton Broth

ABSTRACT The in vitro activity of daptomycin is affected by the concentration of calcium cations in the test medium. Mueller-Hinton broth is currently adjusted to contain 10 to 12.5 mg of magnesium per liter and 20 to 25 mg of calcium per liter, but for testing of daptomycin, greater concentrations of calcium (50 mg/liter) are recommended to better resemble the normal concentration of ionized calcium in human serum. Two levels of calcium were used for broth microdilution tests of 2,789 recent clinical isolates of gram-positive bacterial pathogens. MICs of daptomycin were two- to fourfold lower when the broth contained additional calcium. For most species, however, the percentages of strains that were inhibited by 2.0 μg of daptomycin per ml were essentially identical with the two broth media. Enterococci were the important exception; i.e., 92% were inhibited when tested in calcium-supplemented broth but only 35% were inhibited by 2.0 μg/ml without the additional calcium. This type of information should be considered when selecting criteria for defining in vitro susceptibility to daptomycin.

Effect of Ethanol/Trisodium Citrate Lock on Microorganisms Causing Hemodialysis Catheter-Related Infections

2007 ◽  
Vol 8 (4) ◽  
pp. 262-267 ◽  
Author(s):  
T.A. Takla ◽  
S.A. Zelenitsky ◽  
L.M. Vercaigne
Keyword(s):  
In Vitro Study ◽  
Trisodium Citrate ◽  
Methicillin Resistant ◽  
Hemodialysis Catheter ◽  
E Coli ◽  
Lock Solution ◽  
Mueller Hinton ◽  
Mueller Hinton Broth ◽  
Made In

Purpose This in vitro study tested the effectiveness of a novel 30% ethanol/4% trisodium citrate (TSC) lock solution against the most common pathogens causing hemodialysis catheter-related infections. Methods Clinical isolates of methicillin-resistant Staphylococcus aureus (MRSA) (n=4), methicillin-sensitive S. aureus (MSSA) (n=8), methicillin-resistant Staphylococcus epidermidis (MRSE) (n=8), Pseudomonas aeruginosa (n=4) and Escherichia coli (n=4) were tested in duplicate. Bacterial suspensions of each isolate were made in a control solution of normal saline and Mueller-Hinton broth (MHB), and in a lock solution of ethanol 30%, TSC 4% and MHB. Suspensions were incubated at 37 °C for 48 h. Colony counts were determined from samples collected at t=0 h (before exposure to the ethanol/TSC lock), t=1 h (one hour after exposure to the ethanol/TSC lock), t=24 h and t=48 h. To confirm the absence of viable organisms in the lock solution, the remaining volume at 48 h was filtered through a 0.45 μm filter. The filter was rinsed with 15 mL sterile water and plated on tryptic soy agar (TSA). Results All controls demonstrated significant growth over 48 h. In the lock solutions, initial inocula were reduced to 0 viable colonies by t=1 h (6-log kill), and there was no growth at t=24 and 48 h. Filtering of lock solutions also showed no growth. These results were consistent among duplicates of all isolates. Conclusions The 30% ethanol/4% TSC lock solution consistently eradicated MRSA, MSSA, MRSE, P. aeruginosa and E. coli within 1 h of exposure. Experiments are currently underway to test this novel lock solution on preventing biofilm production by these pathogens.

Laser-Excited Fluorescence of Dityrosine

1995 ◽  
Vol 49 (11) ◽  
pp. 1669-1676 ◽  
Author(s):  
Sahar F. Mahmoud ◽  
Stephen E. Bialkowski
Keyword(s):  
Ferrous Iron ◽  
Ferric Iron ◽  
Iron Concentration ◽  
Fluorescence Emission ◽  
Fluorescence Yield ◽  
Photomultiplier Tube ◽  
Alkaline Solutions ◽  
Array Detector ◽  
Fluorescence Maximum ◽  
Fluorescence Efficiency

In this research, laser-excited fluorescence was examined for sensitive detection of aqueous dityrosine. Samples were excited with a 6.3-mW, 325-nm helium-cadmium laser focused into a small volume-fluorescence cell with a 10-cm lens. The resulting fluorescence emission was collected perpendicular to the excitation and detected with two different schemes. An optical bandpass filter was used with a photomultiplier tube for sensitive quantitative measurement, while a photodiode array detector was used in conjunction with a spectrograph for qualitative characterization of fluorescence emission spectra. Dityrosine detection on the order of 2 × 10−11 M was obtained with the use of the photomultiplier tube with bandpass optical filter. The dityrosine fluorescence yield is found to vary with the solution pH, the relative concentrations of ferric and ferrous iron, and the amount of dissolved oxygen. A maximum fluorescence yield is observed for iron-free, oxygen-free alkaline solutions. Fluorescence quenching by oxygen is a cumulative photolysis effect that diminished fluorescence yield with increased irradiation time. Flowing the solutions minimized photolysis effects in oxygenated solutions. Quenching by ferrous and ferric iron is found to be due primarily to complex formation. The ferrous iron complex appears to have a fluorescence efficiency of ∼20% of the free dityrosine. The ferric iron dityrosine complex appears to have two ferric ions per molecule at low iron concentration. Other complexes may form at different concentrations. Solvent effects on dityrosine absorption and fluorescence spectra were also investigated. A red shift in dityrosine fluorescence maximum was observed in 1 M trichloroacetic acid and in N, N-dimethylformamide. The fluorescence emission maximum was shifted to the blue in acetonitrile and glacial acetic acid. These shifts were attributed to typical solvochromic behavior.

scholarly journals Antimicrobial susceptibility pattern of Corynebacterium striatum.

1996 ◽  
Vol 40 (11) ◽  
pp. 2671-2672 ◽  
Author(s):  
L Martínez-Martínez ◽  
A Pascual ◽  
K Bernard ◽  
A I Suárez
Keyword(s):  
Antimicrobial Susceptibility ◽  
Antimicrobial Agents ◽  
Susceptibility Pattern ◽  
Beta Lactams ◽  
Antimicrobial Susceptibility Pattern ◽  
Mueller Hinton ◽  
Corynebacterium Striatum ◽  
Mueller Hinton Broth

The in vitro activities of 16 antimicrobial agents against 86 strains of Corynebacterium striatum were evaluated by microdilution using cation-adjusted Mueller-Hinton broth. MICs at which 90% of strains were inhibited were 0.06 microgram/ml for teicoplanin, 1 microgram/ml for vancomycin, 0.03 to 8 micrograms/ml for beta-lactams, 8 micrograms/ml for sparfloxacin, 16 micrograms/ml for ciprofloxacin, 16/304 micrograms/ml for co-trimoxazole (trimethoprim-sulfamethoxazole), 64 micrograms/ml for tetracycline, 128 micrograms/ml for gentamicin, and > 128 micrograms/ml for amikacin, erythromycin, and rifampin.

Modeling and Predicting the Growth of Lactic Acid Bacteria in Lightly Preserved Seafood and Their Inhibiting Effect on Listeria monocytogenes

2007 ◽  
Vol 70 (11) ◽  
pp. 2485-2497 ◽  
Author(s):  
OLE MEJLHOLM ◽  
PAW DALGAARD
Keyword(s):  
Lactic Acid ◽  
Listeria Monocytogenes ◽  
Lactic Acid Bacteria ◽  
Chemical Characterization ◽  
Effect Of Temperature ◽  
Inhibiting Effect ◽  
Smoked Salmon ◽  
Boundary Model ◽  
Parameter Values ◽  
Separate Product

A cardinal parameter model was developed to predict the effect of diacetate, lactate, CO2, smoke components (phenol), pH, NaCl, temperature, and the interactions between all parameters on the growth of lactic acid bacteria (LAB) in lightly preserved seafood. A product-oriented approach based on careful chemical characterization and growth of bacteria in ready-to-eat seafoods was used to develop this new LAB growth model. Initially, cardinal parameter values for the inhibiting effect of diacetate, lactate, CO2, pH, and NaCl–water activity were determined experimentally for a mixture of LAB isolates or were obtained from the literature. Next, these values and a cardinal parameter model were used to model the effect of temperature (Tmin) and smoke components (Pmax). The cardinal parameter model was fitted to data for growth of LAB (μmax values) in lightly preserved seafood including cold-smoked and marinated products with different concentrations of naturally occurring and added organic acids. Separate product validation studies of the LAB model resulted in average bias and accuracy factor values of 1.2 and 1.5, respectively, for growth of LAB (μmax values) in lightly preserved seafood. Interaction between LAB and Listeria monocytogenes was predicted by combining the developed LAB model and an existing growth and growth boundary model for the pathogen (O. Mejlholm and P. Dalgaard, J. Food Prot. 70:70–84). The performance of the existing L. monocytogenes model was improved by taking into account the effect of microbial interaction with LAB. The observed and predicted maximum population densities of L. monocytogenes in inoculated lightly preserved seafoods were 4.7 and 4.1 log CFU g−1, respectively, whereas for naturally contaminated vacuum-packed cold-smoked salmon the corresponding values were 0.7 and 0.6 log CFU g−1 when a relative lag time of 4.5 was used for the pathogen.

Tungstenmethyl-Dimethylsilanols Cp(Oc)2(R3P)W—CH2—SiMe2OH (R = Me, Ph) – Synthesis Via Oxygenation or Hydrolysis

2003 ◽  
Vol 58 (1) ◽  
pp. 36-43 ◽  
Author(s):  
Marco Hofmann ◽  
Wolfgang Malisch ◽  
Heike Hupfer ◽  
Martin Nieger
Keyword(s):  
X Ray ◽  
New Compounds ◽  
Hydrolysis Of

Abstract The tungstenmethyl-dimethylsilanols Cp(OC)2(R3P)W-CH2-SiMe2OH [R = Me (6a), Ph (6b)] have been synthesized by oxofunctionalization of the tungstenmethyl-silanes Cp(OC)2(R3P)W- CH2-SiMe2H [R = Me (3a), Ph (3b)] with dimethyldioxirane. 6a has been additionally obtained by Et3N-assisted hydrolysis of the tungstenmethyl-chlorosilane Cp(OC)2(Me3P)W-CH2- SiMe2Cl (5). Compounds 6a,b are stable with respect to self-condensation, but controlled condensation of 6a with Me2Si(H)Cl in the presence of triethylamine has been realized to give the tungstenmethyl-substituted disiloxane Cp(OC)2(Me3P)W-CH2-SiMe2O-SiMe2H (7). The new compounds have been identified IR- and NMR-spectroscopically and, in the case of 3a, by X-ray analysis.

scholarly journals 1588. Delafloxacin Activity Against Staphylococcus aureus with Reduced Susceptibility or Resistance to Methicillin, Vancomycin, Daptomycin, or Linezolid

2019 ◽  
Vol 6 (Supplement_2) ◽  
pp. S579-S580
Author(s):  
Louis D Saravolatz ◽  
Joan Pawlak
Keyword(s):  
Antimicrobial Agents ◽  
Minimal Bactericidal Concentration ◽  
Limited Data ◽  
Methicillin Resistant ◽  
Mueller Hinton ◽  
Spectrum Activity ◽  
Resistant Strains ◽  
Gram Negative Organisms ◽  
Mueller Hinton Broth ◽  
Broad Spectrum Activity

Abstract Background Delafloxacin is a recently approved anionic fluoroquinolone antibiotic with broad-spectrum activity against Gram-positive and Gram-negative organisms. The drug has been approved for patients with acute bacterial skin and skin structure infections including those caused by methicillin-resistant S. aureus. There is limited data available against methicillin-resistant S. aureus blood isolates (MRSABI), vancomycin intermediate strains (VISA), vancomycin-resistant strains (VRSA), daptomycin non-susceptible strains (DNSSA) and linezolid-resistant S. aureus (LRSA). Methods Antimicrobial activity of delafloxacin, levofloxacin, vancomycin, daptomycin, ceftaroline, and linezolid was determined against recent (2016–2018) MRSABI (110), VRSA (15), VISA (35), DNSSA (40), and LRSA (6). Broth microdilution testing using Mueller–Hinton broth was used to determine minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) according to CLSI guidelines. FDA breakpoints were used to determine delafloxacin susceptibility, and CLSI breakpoints were used for all other antibiotics. Results Antimicrobial MIC90 expressed in mg/L and (% susceptible) None of the LRSA were susceptible to delafloxacin or levofloxacin. All strains that were susceptible to the antimicrobial agents above had an MBC that was the same as the MIC or one dilution greater except for linezolid which demonstrated an MBC that was more than eight-fold greater than the MIC. For MRSABI isolates with a levofloxacin MIC ≥ 8 mg/L (55/110) suggesting multiple mutations in the quinolone-resistant determining region, the delafloxacin MIC90 was 1 mg/L with a 36.4% susceptibility rate. Conclusion Delafloxacin demonstrates superior activity to levofloxacin against recent MRSA blood isolates, VISA, VRSA, and DNSSA. Disclosures All authors: No reported disclosures.

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