A Solid Agar Overlay Method for Recovery of Heat-Injured Listeria monocytogenes

2006 ◽  
Vol 69 (2) ◽  
pp. 428-431 ◽  
Author(s):  
ZHINONG YAN ◽  
JOSHUA B. GURTLER ◽  
JEFFREY L. KORNACKI
Keyword(s):  
Listeria Monocytogenes ◽  
Yeast Extract ◽  
Foodborne Pathogens ◽  
Bacterial Cells ◽  
Heat Injury ◽  
Layer Method ◽  
Heat Treated ◽  
Solid Agar ◽  
Overlay Method ◽  
Agar Layer

A solid agar overlay method was developed for recovery of heat-injured Listeria monocytogenes. Presolidified nonselective tryptic soy agar with 0.6% yeast extract (TSAYE, 2% agar) was overlaid on top of solidified modified Oxford agar (MOX). Heat injury of L. monocytogenes was conducted at 58°C for 6 min in a jacketed flask filled with tryptic soy broth. Both noninjured and heat-treated L. monocytogenes cells were plated onto TSAYE, MOX, and TSAYE-MOX plates. No significant differences (P > 0.05) in recovery were found among the three media for noninjured bacterial cells. Recovery of heat-injured L. monocytogenes cells on TSAYE-MOX overlay plates was equivalent to that on the nonselective TSAYE medium, whereas recovery on the selective MOX medium was significantly lower (P < 0.05) compared with both TSAYE and the overlay plates. There were no significant differences (P > 0.05) among the overlay plates prepared 0, 2, 4, 6, 8, 16, and 24 h prior to plating heat-injured bacterial cells. The TSAYE-MOX overlay also allowed differentiation of L. monocytogenes from a mixture of four other types of foodborne pathogens. This solid agar overlay method for recovery of heat-injured L. monocytogenes cells is less time-consuming and less complicated than the conventional overlay-underlay technique and the double overlay modification of the thin agar layer method and may allow for greater laboratory plating efficiencies.

Evaluation of the Thin Agar Layer Method for the Recovery of Pressure-Injured and Heat-Injured Listeria monocytogenes

2014 ◽  
Vol 77 (5) ◽  
pp. 828-831 ◽  
Author(s):  
NICOLAS A. LAVIERI ◽  
JOSEPH G. SEBRANEK ◽  
JOSEPH C. CORDRAY ◽  
JAMES S. DICKSON ◽  
STEPHANIE JUNG ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Yeast Extract ◽  
Cellular Damage ◽  
Growth Media ◽  
Heat Injury ◽  
System Pressure ◽  
Pressure Injury ◽  
A Cell ◽  
Agar Layer ◽  
Antimicrobial Intervention

A sublethally injured bacterial cell has been defined as a cell that survives a stress such as heating, freezing, acid treatment, or other antimicrobial intervention but can repair the cellular damage exerted by the stressor and later regain its original ability to grow. Consequently, sublethally injured cells are not likely to be included in conventional enumeration procedures, which could result in unrealistically low counts unless efforts are made to encourage recovery of the injured cells before enumeration. The objective of this study was to evaluate the use of the thin agar layer (TAL) method for the recovery of pressure-injured and heat-injured Listeria monocytogenes in a tryptic soy broth with 0.6% yeast extract system. Pressure injury consisted of treatment of a culture of mixed L. monocytogenes strains with high hydrostatic pressure at 400 or 600 MPa for 1 s, 2 min, 4 min, or 6 min at a process temperature of 12 ± 2 deg;C. Heat injury consisted of treatment of a culture of mixed L. monocytogenes strains at 60 ± 1°C for 3, 6, or 9 min. Growth media were tryptic soy agar (TSA) with 0.6% yeast extract, modified Oxford medium (MOX), and TAL, which consisted of a 7-ml layer of TSA overlaid onto solidified MOX. Counts of viable L. monocytogenes on TAL were higher than those on MOX in the heat-injury experiment but not in the pressure-injury experiment. Therefore, the effectiveness of the TAL method may be specific to the type of injury applied to the microorganism and should be investigated in a variety of cellular injury scenarios.

Evaluation of Thin Agar Layer Method for Recovery of Acid-Injured Foodborne Pathogens†

2001 ◽  
Vol 64 (7) ◽  
pp. 1067-1071 ◽  
Author(s):  
V. C. H. WU ◽  
D. Y. C. FUNG ◽  
D. H. KANG ◽  
L. K. THOMPSON
Keyword(s):  
Foodborne Pathogens ◽  
Petri Dish ◽  
Selective Medium ◽  
Escherichia Coli O157 ◽  
Selective Media ◽  
Layer Method ◽  
Significant Difference ◽  
Selective Agents ◽  
One Step ◽  
Agar Layer

The thin agar layer (TAL) method of Kang and Fung was used to enumerate acid-injured foodborne pathogens. This method involves overlaying 14 ml of nonselective medium (tryptic soy agar [TSA]) onto a prepoured and solidified pathogen-specific, selective medium in a petri dish. After surface plating, injured cells resuscitated and grew on TSA during the first few hours of incubation; then, the selective agents from the selective medium diffused to the top layer, interacted with the recovered microorganisms, and started to produce typical reactions. Foodborne pathogens were exposed to 2% acetic acid for 1, 2, or 4 min, and the recovery rate with the TAL method was compared with the rate of TSA and pathogen-specific, selective media. No significant difference occurred between TSA and TAL (P > 0.05) for enumeration of acid-injured Escherichia coli O157:H7, Salmonella Typhimurium, Staphylococcus aureus, and Yersinia enterocolitica, and both recovered significantly higher numbers than the selective medium for each respective pathogen (P < 0.05). For recovery of acid-injured Listeria monocytogenes, no difference (P > 0.05) occurred among TSA, TAL, and selective media. However, fewer cells were recovered in the selective media. The TAL method is a one-step, convenient procedure for recovery of acid-injured cells.

Use of Conventional Media and Thin Agar Layer Method for Recovery of Foodborne Pathogens from Pressure-treated Poultry Products

2003 ◽  
Vol 68 (7) ◽  
pp. 2321-2324 ◽  
Author(s):  
J. Yuste ◽  
M. Capellas ◽  
R. Pla ◽  
S. Llorens ◽  
D.Y.C. Fung ◽  
...  
Keyword(s):  
Foodborne Pathogens ◽  
Poultry Products ◽  
Layer Method ◽  
Agar Layer

Changes in Aerobic Plate and Escherichia coli–Coliform Counts and in Populations of Inoculated Foodborne Pathogens on Inshell Walnuts during Storage

2016 ◽  
Vol 79 (7) ◽  
pp. 1143-1153 ◽  
Author(s):  
JOHN C. FRELKA ◽  
GORDON R. DAVIDSON ◽  
LINDA J. HARRIS
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Relative Humidity ◽  
Low Temperature ◽  
Foodborne Pathogens ◽  
Limit Of Detection ◽  
Sampling Time ◽  
E Coli ◽  
Plate Counts ◽  
Forced Air

ABSTRACT After harvest, inshell walnuts are dried using low-temperature forced air and are then stored in bins or silos for up to 1 year. To better understand the survival of bacteria on inshell walnuts, aerobic plate counts (APCs) and Escherichia coli–coliform counts (ECCs) were evaluated during commercial storage (10 to 12°C and 63 to 65% relative humidity) over 9 months. APCs decreased by 1.4 to 2.0 log CFU per nut during the first 5 months of storage, and ECCs decreased by 1.3 to 2.2 log CFU per nut in the first month of storage. Through the remaining 4 to 8 months of storage, APCs and ECCs remained unchanged (P > 0.05) or decreased by <0.15 log CFU per nut per month. Similar trends were observed on kernels extracted from the inshell walnuts. APCs and ECCs were consistently and often significantly higher on kernels extracted from visibly broken inshell walnuts than on kernels extracted from visibly intact inshell walnuts. Parameters measured in this study were used to determine the survival of five-strain cocktails of E. coli O157:H7, Listeria monocytogenes, and Salmonella inoculated onto freshly hulled inshell walnuts (~8 log CFU/g) after simulated commercial drying (10 to 12 h; 40°C) and simulated commercial storage (12 months at 10°C and 65% relative humidity). Populations declined by 2.86, 5.01, and 4.40 log CFU per nut for E. coli O157:H7, L. monocytogenes, and Salmonella, respectively, after drying and during the first 8 days of storage. Salmonella populations changed at a rate of −0.33 log CFU per nut per month between days 8 and 360, to final levels of 2.83 ± 0.79 log CFU per nut. E. coli and L. monocytogenes populations changed by −0.17 log CFU per nut per month and −0.26 log CFU per nut per month between days 8 and 360, respectively. For some samples, E. coli or L. monocytogenes populations were below the limit of detection by plating (0.60 log CFU per nut) by day 183 or 148, respectively; at least one of the six samples was positive at each subsequent sampling time by either plating or by enrichment.

Evaluation of Gaseous Chlorine Dioxide as a Sanitizer for Killing Salmonella, Escherichia coli O157:H7, Listeria monocytogenes, and Yeasts and Molds on Fresh and Fresh-Cut Produce

2005 ◽  
Vol 68 (6) ◽  
pp. 1176-1187 ◽  
Author(s):  
KAYE V. SY ◽  
MELINDA B. MURRAY ◽  
M. DAVID HARRISON ◽  
LARRY R. BEUCHAT
Keyword(s):  
Escherichia Coli ◽  
Listeria Monocytogenes ◽  
Chlorine Dioxide ◽  
Foodborne Pathogens ◽  
Escherichia Coli O157 ◽  
Gaseous Chlorine Dioxide ◽  
Sensory Qualities ◽  
A Cell ◽  
Fresh Cut ◽  
Yeasts And Molds

Gaseous chlorine dioxide (ClO2) was evaluated for effectiveness in killing Salmonella, Escherichia coli O157:H7, and Listeria monocytogenes on fresh-cut lettuce, cabbage, and carrot and Salmonella, yeasts, and molds on apples, peaches, tomatoes, and onions. Inoculum (100 μl, ca. 6.8 log CFU) containing five serotypes of Salmonella enterica, five strains of E. coli O157:H7, or five strains of L. monocytogenes was deposited on the skin and cut surfaces of fresh-cut vegetables, dried for 30 min at 22°C, held for 20 h at 4°C, and then incubated for 30 min at 22°C before treatment. The skin surfaces of apples, peaches, tomatoes, and onions were inoculated with 100 μl of a cell suspension (ca. 8.0 log CFU) containing five serotypes of Salmonella, and inoculated produce was allowed to dry for 20 to 22 h at 22°C before treatment. Treatment with ClO2 at 4.1 mg/liter significantly (α = 0.05) reduced the population of foodborne pathogens on all produce. Reductions resulting from this treatment were 3.13 to 4.42 log CFU/g for fresh-cut cabbage, 5.15 to 5.88 log CFU/g for fresh-cut carrots, 1.53 to 1.58 log CFU/g for fresh-cut lettuce, 4.21 log CFU per apple, 4.33 log CFU per tomato, 1.94 log CFU per onion, and 3.23 log CFU per peach. The highest reductions in yeast and mold populations resulting from the same treatment were 1.68 log CFU per apple and 2.65 log CFU per peach. Populations of yeasts and molds on tomatoes and onions were not significantly reduced by treatment with 4.1 mg/liter ClO2. Substantial reductions in populations of pathogens on apples, tomatoes, and onions but not peaches or fresh-cut cabbage, carrot, and lettuce were achieved by treatment with gaseous ClO2 without markedly adverse effects on sensory qualities.

Effect of pH on the Minimum Inhibitory Concentration of Monolaurin Against Listeria monocytogenes1

1992 ◽  
Vol 55 (6) ◽  
pp. 449-452 ◽  
Author(s):  
DEOG-HWAN OH ◽  
DOUGLAS L. MARSHALL
Keyword(s):  
Listeria Monocytogenes ◽  
Minimum Inhibitory Concentration ◽  
Yeast Extract ◽  
Inhibitory Concentration ◽  
Ph Value ◽  
Potassium Sorbate ◽  
Butylated Hydroxyanisole ◽  
Tertiary Butylhydroquinone ◽  
Effect Of Ph ◽  
Glycerol Monolaurate

The effect of pH on the minimum inhibitory concentration (MIC) of glycerol monolaurate (monolaurin) against four strains of Listeria monocytogenes at 35°C in tryptic soy broth supplemented with 0.6% yeast extract was investigated. Our results demonstrate that the MIC of monolaurin was lower than MIC values reported for other common food antimicrobials such as potassium sorbate, tertiary butylhydroquinone, propyl paraben, and butylated hydroxyanisole. The MIC of monolaurin was reduced by decreasing the pH value of the medium. A 3-fold MIC reduction occurred when the pH decreased from pH 7.0 (10 μg/ml) to pH 5.0 (3 μg/ml).

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