Spray Method for Recovery of Heat-Injured Salmonella Typhimurium and Listeria monocytogenes

2012 ◽  
Vol 75 (10) ◽  
pp. 1867-1872 ◽  
Author(s):  
KYEONG-HWAN BACK ◽  
SANG-OH KIM ◽  
KI-HWAN PARK ◽  
MYUNG-SUB CHUNG ◽  
DONG-HYUN KANG
Keyword(s):  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Recovery Rate ◽  
Skim Milk ◽  
Selective Media ◽  
Spray Method ◽  
Selective Agar ◽  
Overlay Method ◽  
Peptone Water ◽  
Pathogen Recovery

Selective agar is inadequate for supporting recovery of injured cells. During risk assessment of certain foods, both injured and noninjured cells must be enumerated. In this study, a new method (agar spray method) for recovering sublethally heat-injured microorganisms was developed and used for recovery of heat-injured Salmonella Typhimurium and Listeria monocytogenes. Molten selective agar was applied as an overlay to presolidified nonselective tryptic soy agar (TSA) by spray application. Heat-injured cells (55°C for 10 min in 0.1% peptone water or 55°C for 15 min in sterilized skim milk) were inoculated directly onto solidified TSA. After a 2-h incubation period for cell repair, selective agar was applied to the TSA surface with a sprayer, and the plates were incubated. The recovery rate for heat-injured Salmonella Typhimurium and L. monocytogenes with the spray method was compared with the corresponding rates associated with TSA alone, selective media alone, and the conventional overlay method (selective agar poured on top of resuscitated cells grown on TSA and incubated for 2 h). No significant differences (P > 0.05) were found in pathogen recovery obtained with TSA, the overlay method, and the spray method. However, a lower recovery rate (P < 0.05) was obtained for isolation of injured cells on selective media. Overall, these results indicate that the agar spray method is an acceptable alternative to the conventional overlay method and is a simpler and more convenient approach to recovery and detection of injured cells.

Transfer Volume-Dependent Recovery of Salmonella from Minced Meat

1987 ◽  
Vol 50 (7) ◽  
pp. 584-586 ◽  
Author(s):  
S. KAFEL ◽  
E. POGORZELSKA
Keyword(s):  
Salmonella Typhimurium ◽  
Broth Culture ◽  
Selective Media ◽  
Selective Enrichment ◽  
Salmonella Choleraesuis ◽  
Selective Agar ◽  
Peptone Water ◽  
Pure Cultures ◽  
The Mean ◽  
Minced Meat

Six hundred 25-g samples of ground beef were divided into 3 groups of 200 each and 1 drop of a Salmonella broth culture was added to each sample. After storage at −27°C for 3–4 months, the samples were defrosted and blended with 225 ml of buffered peptone water. Ten ml of each suspension was preenriched at 37°C for 20 h and 10-fold dilutions of the material were made. One ml each of the preenriched culture and dilutions of 10−2, 10−4, 10−6, and 10−8 were transferred to selective enrichment media, and subsequently streaked onto selective agar plates. The mean percentage of Salmonella-positives obtained from all combinations of the selective media in relation to undiluted preenriched material and its 10−2, 10−4, 10−6, and 10−8 dilutions were for Salmonella typhimurium 70, 81, 84, 37, and 2, for Salmonella choleraesuis 64, 78, 66, 30, and 2, and for Salmonella anatum 60, 84, 75, 40, and 1, respectively. Colonies originating from diluted samples, particularly 10−4 and further dilutions, usually represented pure cultures of salmonellae, but from undiluted material were frequently accompanied or outgrown by concomitant bacteria.

Evaluation of Nisin-Coated Cellulose Casings for the Control of Listeria monocytogenes Inoculated onto the Surface of Commercially Prepared Frankfurters†‡

2004 ◽  
Vol 67 (5) ◽  
pp. 1017-1021 ◽  
Author(s):  
JOHN B. LUCHANSKY ◽  
JEFFREY E. CALL
Keyword(s):  
Listeria Monocytogenes ◽  
Surface Area ◽  
Internal Surface ◽  
Refrigerated Storage ◽  
Internal Surface Area ◽  
Appreciable Increase ◽  
Potassium Lactate ◽  
Selective Agar ◽  
Peptone Water ◽  
Sodium Diacetate

Commercially prepared frankfurters were formulated with and without ~1.4% potassium lactate and 0.1% sodium diacetate and were subsequently processed in cellulose casings coated with and without nisin (~50,000 IU per square inch of internal surface area) to control the outgrowth of Listeria monocytogenes during refrigerated storage. The frankfurters were inoculated with ~5 log CFU per package of a five-strain mixture of L. monocytogenes and then vacuum sealed before being stored at 4° C for 60 to 90 days. Surviving organisms were recovered and enumerated by rinsing each package with 18 ml of sterile 0.1% peptone water and plating onto MOX selective agar. The data for each of two trials were averaged. In packages that contained frankfurters formulated with potassium lactate and sodium diacetate and prepared in nisin-coated casings, L. monocytogenes levels decreased by 1.15 log CFU per package after 90 days of storage. L. monocytogenes levels decreased by 0.95 log CFU per package in frankfurters that were prepared in casings that were not coated with nisin. In packages of frankfurters that were formulated without potassium lactate and sodium diacetate and prepared in nisin-coated casings, L. monocytogenes levels decreased by 0.88 log CFU per package after 15 days of storage but then increased appreciablythereafter over a 60-day period of refrigerated storage. There was also an appreciable increase in pathogen numbers during 60 days of storage in otherwise similar frankfurters formulated without potassium lactate and sodium diacetate prepared in casings that were not coated with nisin. These data confirm that potassium lactate and sodium diacetate display listeriostatic activity as an ingredient of commercial frankfurters. These data also establish that cellulose casings coated with nisin display only moderate antilisterial activity in vacuum-sealed packages of commercially prepared frankfurters during storage at 4° C.

Comparison of Selective Direct Plating Media for Enumeration and Recovery of L. monocytogenes from Cold-process (smoked) Fish

1992 ◽  
Vol 55 (11) ◽  
pp. 905-909 ◽  
Author(s):  
ROHINEE N. PARANJPYE ◽  
GRETCHEN A. PELROY ◽  
MARK E. PETERSON ◽  
FRANK T. POYSKY ◽  
PAUL J. HOLLAND ◽  
...  
Keyword(s):  
Listeria Monocytogenes ◽  
Lithium Chloride ◽  
Indicator System ◽  
Water Phase ◽  
Direct Plating ◽  
Selective Media ◽  
Smoked Fish ◽  
Smoked Salmon ◽  
Selective Agar ◽  
Added Salt

Three selective media were evaluated for direct plating recovery and enumeration of Listeria monocytogenes in the presence of high levels of a variety of microorganisms occurring on cold-process (smoked) salmon products. Sliced salmon was brined to contain either no added salt, 3, or 5% water-phase NaCl, or 3 or 5% NaCl plus 140 ppm NaNO2. The slices were packaged in oxygen-permeable film or sealed under vacuum in oxygen-impermeable film, and stored at 10°C or 5°C until total microbial loads reached 106 to 109 CFU/g. Oxford formulation of Listeria selective agar and Lee's modification of Listeria selective agar achieved quantitative recovery of 102 cells per ml of L. monocytogenes strain Scott A in the presence of diluted slurries of these fish containing 104 to 108 CFU/ml of background organisms. A modification of lithium chloride-phenylethanol-moxalactam agar containing an iron-esculin indicator system sometimes failed because of interfering growth by the background microflora.

scholarly journals Recovery and Enumeration of Staphylococcus aureus by the Selective Agar

Fine Focus ◽  
2016 ◽  
Vol 2 (1) ◽  
pp. 51-59
Author(s):  
Jill Bange ◽  
Emily Brumfield ◽  
Alysha L. Ellison
Keyword(s):  
Staphylococcus Aureus ◽  
Food Processing ◽  
Recovery Period ◽  
False Negative ◽  
Selective Medium ◽  
Bacterial Cells ◽  
Human Immune System ◽  
Selective Media ◽  
Selective Agar ◽  
Overlay Method

Staphylococcus aureus isan example of a commensal bacterium responsible for emesis, acute diarrheal syndrome, and sepsis. S. aureus often must be isolated from patient samples in a clinical setting or from food samples during food processing in an industrial setting, although these bacterial cells may be injured by the human immune system or by food processing measures. Therefore, injured cells may not be fully recovered on media selective for S. aureus and enumeration (e.g., CFU/mL) may not reflect the true concentration of the original sample. The objective of this study was to determine whether the selective agar overlay method of recovery is more sensitive, selective, and time-effective for enumeration of artificially injured S. aureus cultures when compared to more traditional techniques. The selective agar overlay method involves pour plating S. aureus in non-selective medium, allowing the sample to incubate for a four hour recovery period, and then overlaying selective medium over the non-selective medium. Artificial injury of S. aureus cells was accomplished by treatment with carvacrol, an extract from oil of oregano. Our results indicated that carvacrol-injured S. aureus cells were recovered by the selective agar overlay at the same concentration as recovery on non-selective media, and at a significantly higher concentration than recovery on selective media. This method allows for more rapid and accurate diagnoses, and may be more cost-effective due to the reduction or elimination of false negative results.

Attachment of Listeria monocytogenes and Salmonella typhimurium to Stainless Steel and Buna-N in the Presence of Milk and Individual Milk Components

1993 ◽  
Vol 56 (6) ◽  
pp. 479-484 ◽  
Author(s):  
DAVID M. HELKE ◽  
EILEEN B. SOMERS ◽  
AMY C. L. WONG
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Dairy Industry ◽  
Skim Milk ◽  
Whole Milk ◽  
Milk Components ◽  
Phosphate Buffered Saline ◽  
Β Lactoglobulin

The effects of milk and individual milk components on the attachment of Listeria monocytogenes and Salmonella typhimurium to two commonly used materials in the dairy industry were studied. Attachment of both organisms to stainless steel and Buna-N was significantly inhibited by the presence of skim, 2%, whole, or chocolate 2% milk compared to the phosphate-buffered saline (PBS) control. The addition of individual milk components, casein, α-lactalbumin, and β-lactoglobulin to the attachment menstruum significantly reduced attachment. Pretreating surfaces with milk and milk components for 1 h prior to attachment in PBS gave similar results. The presence of lactose did not affect attachment of either organism; however, attachment of S. typhimurium was significantly decreased on pretreated Buna-N. Cells of either organism pretreated with skim milk or β-lactoglobulin prior to attachment in PBS showed significantly less attachment than untreated cells. Pretreating S. typhimurium cells with casein had no effect on attachment to stainless steel. Pretreatment of S. typhimurium with lactose increased attachment to both surfaces while pretreatment had no effect on L. monocytogenes. Attachment of both organisms was significantly reduced in diluted whole milk. Both organisms attached significantly less to surfaces soiled with one or more layers of whole milk.

Growth Media and Surface Conditioning Influence the Adherence of Pseudomonas fragi, Salmonella typhimurium, and Listeria monocytogenes Cells to Stainless Steel

1997 ◽  
Vol 60 (9) ◽  
pp. 1034-1037 ◽  
Author(s):  
SCOTT K. HOOD ◽  
EDMUND A. ZOTTOLA
Keyword(s):  
Stainless Steel ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Skim Milk ◽  
Epifluorescence Microscopy ◽  
Growth Media ◽  
Pseudomonas Fragi ◽  
Acridine Orange Staining ◽  
Surface Conditioning ◽  
Food Contact Surfaces

Microorganisms have been shown to adhere to food-contact surfaces and may provide a route for the contamination of processed food. To better understand this phenomenon, the effects of growth media and surface conditioning on the adherence of Pseudomonas fragi, Salmonella typhimurium and Listeria monocytogenes cells to stainless steel were studied. The microorganisms were grown in tryptic soy broth (TSB), 1% reconstituted skim milk (RSM) and RSM with 1% sucrose (RSM + S). Stainless-steel surfaces were conditioned by immersion in growth media for 1 h and then were rinsed in phosphate-buffered saline (PBS) prior to the adherence assay. After growing in each medium, cells were harvested, resuspended in PBS, and then allowed to contact the stainless steel for 30 min. Adherence was quantified by acridine orange-staining the cells and viewing under epifluorescence microscopy. Growth media had little influence on adherence to stainless steel that had not been preconditioned. P. fragi and L. monocytogenes cells adhered in the highest numbers when grown in RSM plus sucrose. S. typhimurium cells showed the highest level of adherence when grown in TSB. Analysis of variance yielded P values of less than 0.01, indicating that both growth media and surface conditioning were significant in the level of adherence observed.

Growth of Listeria monocytogenes in the Presence of Pseudomonas fluorescens at 7 or 13°C in Skim Milk

1989 ◽  
Vol 52 (12) ◽  
pp. 852-855 ◽  
Author(s):  
SEHAM A. FARRAG ◽  
ELMER H. MARTH
Keyword(s):  
Listeria Monocytogenes ◽  
Pseudomonas Fluorescens ◽  
Incubation Period ◽  
Skim Milk

Autoclaved samples of skim milk were inoculated with Listeria monocytogenes (strain Scott A, California or V7), Pseudomonas fluorescens (strain P26 or B52), or a combination of L. monocytogenes plus P. fluorescens, and incubated at 7 or 13°C for 8 weeks. McBride Listeria Agar was used to determine populations of L. monocytogenes (at 0, 7, 14, 28, 42, or 56 d), and Pseudomonas isolation agar to enumerate P. fluorescens. Growth of L. monocytogenes was somewhat enhanced after 7 d of incubation at 7 but not at 13°C in the presence of pseudomonads. However, after 14 d and until the end of the incubation period (56 d), slight inactivation of L. monocytogenes in the presence of P. fluorescens was observed. L. monocytogenes did not affect growth or survival of P. fluorescens; also, no marked changes in pH of the milk were caused either by L. monocytogenes alone or by L. monocytogenes plus P. fluorescens.

scholarly journals Autophagy—A Story of Bacteria Interfering with the Host Cell Degradation Machinery

Pathogens ◽  
2021 ◽  
Vol 10 (2) ◽  
pp. 110
Author(s):  
Anna K. Riebisch ◽  
Sabrina Mühlen ◽  
Yan Yan Beer ◽  
Ingo Schmitz
Keyword(s):  
Escherichia Coli ◽  
Staphylococcus Aureus ◽  
Immune Response ◽  
Mycobacterium Tuberculosis ◽  
Listeria Monocytogenes ◽  
Salmonella Typhimurium ◽  
Innate Immune ◽  
Intracellular Pathogens ◽  
Response Mechanism ◽  
Pathogenic Escherichia Coli

Autophagy is a highly conserved and fundamental cellular process to maintain cellular homeostasis through recycling of defective organelles or proteins. In a response to intracellular pathogens, autophagy further acts as an innate immune response mechanism to eliminate pathogens. This review will discuss recent findings on autophagy as a reaction to intracellular pathogens, such as Salmonella typhimurium, Listeria monocytogenes, Mycobacterium tuberculosis, Staphylococcus aureus, and pathogenic Escherichia coli. Interestingly, while some of these bacteria have developed methods to use autophagy for their own benefit within the cell, others have developed fascinating mechanisms to evade recognition, to subvert the autophagic pathway, or to escape from autophagy.

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