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SURVIVAL AND GROWTH OF LISTERIA MONOCYTOGENES IN CHOPPED GREEN AND RED BELL PEPPERS USING PREDICTIVE MODELING
Title
SURVIVAL AND GROWTH OF LISTERIA MONOCYTOGENES IN CHOPPED GREEN AND RED BELL PEPPERS USING PREDICTIVE MODELING
Creator
Zhang, L1jie
Date
2015, 2015-07
Description
Listeria monocytogenes (L. monocytogenes) is a Gram-positive pathogenic organism and the causative agent of human and animal listeriosis....
Show moreListeria monocytogenes (L. monocytogenes) is a Gram-positive pathogenic organism and the causative agent of human and animal listeriosis. Listeriosis is a gastrointestinal or invasive systemic illness resulting from consumption of contaminated food products, mainly cheese, deli meats, and fresh produce, by L. monocytogenes. In recent years, several foodborne outbreaks have been reported that were associated with fresh produce, such as cantaloupe, celery and sprouts. Temperature is considered a major factor that affects L. monocytogenes growth during storage. The proliferation of L. monocytogenes varies on different produce items based on storage temperature. In this study, the persistence and population dynamics of three L. monocytogenes strains, LS806 (cheese isolate), LS810 (cantaloupe isolate) and LS808 (celery isolate) were evaluated by incubating inoculated fresh-cut green bell pepper and red bell pepper at various temperatures (5oC, 10oC, and 25oC) for 14 days. To assess the risk of L. monocytogenes in these fresh-cut vegetable items, a primary predictive model was fitted for L. monocytogenes growth data using DMFit. Green bell pepper had significantly (P<0.05) higher pH and aw, and higher amounts of yeast and mold and Enterobacteriaceae than did red bell pepper. In green bell pepper, all three strains showed no significant difference (P>0.05) in growth rate when incubated at the same temperature. In red bell peppers, LS808 had the highest (P<0.05) growth rate at both 5°C and 25°C out of the three strains. All of the three strains grew significantly faster (P<0.01) at 25°C than either 5°C or 10°C in both green and red bell peppers. All three strains obtained less than 1 log10 growth increase after incubating at 25°C for 6 hours on pre-chilled produce. Some strains (LS806 and LS810) significantly increased (P<0.01) during two 5-hour 25°C incubations, but did not reach 1 log10 growth increase. The results indicate that L. monocytogenes not only persists, but also grows in chopped green and red bell peppers at 5, 10, and 25°C, and strains grew faster at the higher temperature (25°C). Data obtained could be further evaluated for determining whether Time/temperature control for safety (TCS) designation should be applied to chopped green and red bell peppers.
M.S. in Food Safety and Technology, July 2015
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THE TRANSCRIPTIONAL RESPONSE OF ARTIFICIALLY-INOCULATED LISTERIA MONOCYTOGENES DURING THE MANUFACTURE OF UNPASTEURIZED GOUDA CHEESE
Title
THE TRANSCRIPTIONAL RESPONSE OF ARTIFICIALLY-INOCULATED LISTERIA MONOCYTOGENES DURING THE MANUFACTURE OF UNPASTEURIZED GOUDA CHEESE
Creator
Carstens, Christina K.
Date
2017, 2017-05
Description
Listeriosis outbreaks indicate that L. monocytogenes contamination is an issue for various food types, such as unpasteurized (raw) cheese....
Show moreListeriosis outbreaks indicate that L. monocytogenes contamination is an issue for various food types, such as unpasteurized (raw) cheese. Current regulation prevents the interstate sale and distribution of raw cheese and mandates a ≥60 day aging period at ≥2˚C to ensure product safety; yet studies demonstrate that pathogens can persist during aging. Environmental stress can alter the transcriptomic profiles of pathogens; however, these surveys are rarely conducted in food matrices. This study aimed to assess the transcriptomic profiles of L. monocytogenes strain F2365 during environmental stressors inherent throughout cheesemaking. First, quantitative polymerase chain reaction (qPCR) was used to monitor transcription levels of nine L. monocytogenes genes involved in virulence, stress response, energy transport, and metabolism during osmotic stress (11% NaCl) and varying temperature/time (5˚C 24 h, 25˚C 24 h, 38˚C 30 min) conditions in Brain Heart Infusion (BHI) broth, raw milk, and pasteurized milk. Generally, the genes prfA, lmo1381, lmo0963, and lmo1875 were down-regulated, whereas the genes lmo1864, lmo0914, lmo0348, lmo1428, and lmo1264 were up-regulated. Virulence gene, prfA, was most down-regulated when L. monocytogenes was grown in raw milk with salt at 25˚C (4774.72±838.14 fold; relative to 24 h growth at 37˚C). The stress response gene lmo0914, encoding σB, was most up-regulated when L. monocytogenes was grown in BHI at 25˚C with salt (14.61±7.72 fold). Additionally, transcription levels of the nine genes were assessed at points during the laboratory-scale manufacture of Gouda cheese made with raw milk artificially-inoculated with L. monocytogenes via qPCR. Similar differential regulation for both prfA and lmo0914 in L. monocytogenes was observed during cheesemaking. The gene lmo1864, encoding a putative pore-forming hemolysin, was up-regulated throughout the cheesemaking process, but was most up-regulated after stirring the curd (449.81±432.53 fold). Ultimately, these results indicate that the lmo1864 gene may play a role in L. monocytogenes survival during cheesemaking. Methods developed in this study can be used to assess the risk of L. monocytogenes, not only during cheesemaking, but during the ≥60-day aging process. Overall, these results contribute to the understanding of L. monocytogenes survival mechanisms during the cheesemaking process.
M.S. in Food Safety and Technology, May 2017
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SYNERGISTIC EFFECT OF FATTY ACIDS AND NISIN IN INHIBITING PERSISTER AND BIOFILM OF LISTERIA MONOCYTOGENES
Title
SYNERGISTIC EFFECT OF FATTY ACIDS AND NISIN IN INHIBITING PERSISTER AND BIOFILM OF LISTERIA MONOCYTOGENES
Creator
Zhou, Jiacheng
Date
2019
Description
A foodborne pathogen Listeria monocytogenes causes a life-threatening listeriosis in humans after eating contaminated food. The FDA-approved...
Show moreA foodborne pathogen Listeria monocytogenes causes a life-threatening listeriosis in humans after eating contaminated food. The FDA-approved antimicrobial peptide nisin has been used to prevent contamination of food product from Gram-positive pathogens including L. monocytogenes. However, the formation of biofilms and persisters (i.e., metabolically dormant bacterial population) has resulted in the failure of nisin treatment. Fatty acids, which have been known to exhibit antimicrobial activities, are widely used for therapeutics, food preservation, and agriculture. Previously, we found that two fatty acid compounds lauric acids and N-tridecanoic acids are effective in inhibiting biofilms and persister formation of Gram-negative pathogens. In this study, we investigate whether the fatty acid treatment in combination with nisin promotes inactivation of L. monocytogenes, especially biofilms and persisters. The fatty acid-only treatment reduced the level of biofilms and persisters, while nisin-only treatment resulted in the development of resistant population of L. monocytogenes ATCC19115 strain. However, the co-treatment of the fatty acid and nisin synergistically enhanced the killing of L. monocytogenes by significantly decreasing the number of survived cells and inhibiting biofilms. These results are particularly important in improving food safety in that the food-grade fatty acids can be applied to repress the occurrence of resistant mechanisms of foodborne pathogens by inhibiting biofilm and persister cell formation.
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EVALUATION OF LISTERIA MONOCYTOGENES ENRICHMENT AND COMPOSITING PROTOCOLS FROM ENVIRONMENTAL SAMPLES
Title
EVALUATION OF LISTERIA MONOCYTOGENES ENRICHMENT AND COMPOSITING PROTOCOLS FROM ENVIRONMENTAL SAMPLES
Creator
Eckert, Christine
Date
2019
Description
Environmental sampling in a food production plant is routinely conducted using devices, such as sponges or swabs, to verify cleaning...
Show moreEnvironmental sampling in a food production plant is routinely conducted using devices, such as sponges or swabs, to verify cleaning procedures and determine if any foodborne pathogens, such as Listeria monocytogenes (L. monocytogenes), are present. The devices used for environmental monitoring are enriched to improve pathogen detection. This study aims to 1) compare the limit of detection (LOD) of L. monocytogenes of two U.S. Food and Drug Administration (FDA) enrichment procedures (i.e., Bacteriological Analytical Manual (BAM) and Compliance Document) with and without food matrix, and to 2) assess the number of samples which can be wet and dry composited without loss of sensitivity from stainless steel. To compare the LOD of L. monocytogenes using UVM and BLEB, three inoculation levels (0.27±0.07, 0.59±0.05, and 1.00±0.15 CFU per 225 mL enrichment) with 30 enrichments each were used. Results showed that there was no significant difference between the number of samples where L. monocytogenes was detected for UVM and BLEB at any of the three inoculation levels. However, the limit of detection (LOD95%) for UVM/Fraser was higher than that of BLEB (2.13 and 1.44 CFU/mL, respectively). For wet compositing, 1.24±0.34 CFU of L. monocytogenes was inoculated into 45 enrichments of UVM or BLEB without food matrix and 7.2±0.18 CFU of L. monocytogenes was inoculated into 30 enrichments of UVM or BLEB with 4.13±0.12 log CFU of native microflora from Romaine lettuce wash (RLW). Secondary composite enrichments in Fraser broth were conducted at each of four different ratios: 1:1 (1 positive:1 negative), 1:2 (1 positive: 2 negative), 1:4 (1 positive: 4 negative), and 1:7 (1 positive:7 negative). There was no significant difference between the number of samples where L. monocytogenes was detected between BLEB and UVM with or without food matrix at any of the composite ratios. When comparing wet and dry compositing enrichments from stainless steel, 10.16 × 10.16 cm areas on stainless steel plates were inoculated with 464±22 CFU (2.67±0.24 log CFU) L. monocytogenes, dried for 24 h, and sponges were used to swab the surface of the plates. The sponges were then composited (into primary enrichments for dry compositing) or the secondary enrichments were composited (for wet compositing). Compositing was conducted with RLW containing 4.13±0.02 log CFU of background microflora. There was no significant difference between the number of samples where L. monocytogenes was detected for BLEB and UVM when comparing dry or wet compositing at any of the composite ratios tested. Results of this thesis will aid in determining if compositing of environmental samples is an option when L. monocytogenes is the target pathogen.
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Growth kinetics of Salmonella enterica and Listeria monocytogenes during rehydration of dehydrated corn and subsequent storage
Title
Growth kinetics of Salmonella enterica and Listeria monocytogenes during rehydration of dehydrated corn and subsequent storage
Creator
Mate, Madhuri
Date
2022
Description
Dehydrated vegetables, including corn, are often used in restaurants and retail grocers. They do not support the growth of pathogens as their...
Show moreDehydrated vegetables, including corn, are often used in restaurants and retail grocers. They do not support the growth of pathogens as their moisture content is very low. After rehydration, these food products attain high water activity values suitable with neutral pH for the survival and proliferation of foodborne pathogens, including Salmonella enterica and Listeria monocytogenes. The purpose of the study was to examine the extent to which dehydrated corn supports the growth of S. enterica and L. monocytogenes during rehydration at 5 or 25°C water and following storage at 5, 10, and 25°C temperatures at 1, 3, 5 and 7 d intervals. Fresh corn was dehydrated at 60°C for 24 h. Dehydrated corn was inoculated with a 4-strain cocktail of either S. enterica or rifampicin-resistant L. monocytogenes, resulting in 4 log CFU/g, and held at ambient temperature for 24 h. This corn was then rehydrated using either 5 or 25°C water for 24 h. Throughout rehydration, corn samples were removed at intervals and enumerated. To enumerate S. enterica and L. monocytogenes, the samples were homogenized with BPB and BLEB respectively and cultivated on TSAYE with overlaid XLD or BHIARif200, respectively. Rehydrated corn was then stored at 5, 10, or 25°C and enumerated at intervals 1,3,5 and 7 d. Triplicate samples were assessed at each timepoint and three independent experiments were conducted for each rehydration water temperature. Growth rates were determined by DMFit and statistically analyzed using Student t-test. A p-value ≤0.05 was considered significant. Overall the growth rate of S. enterica was higher when rehydrated in 5°C water temperature and then stored at 25°C and was determined to be 0.61 ± 0.23 log CFU/g per d. This timepoint was also the shortest time required to increase by 1 log which was: 1.64 d, i.e. 39 h. For L. monocytogenes, the 25°C water rehydration showed the fastest growth rate when stored at 25°C. It took only 1.58 d or 37.8 h for 1 log increase in the population. After 5°C water rehydration of corn the highest populations of mesophilic bacteria and yeasts and molds were observed for 25°C storage ranging from 8.43 to 9.39 log CFU/g and 4.75 to 7.87 log CFU/g, respectively. After 25°C water rehydration, the highest population of mesophilic bacteria, 8.88 log CFU/g, was observed at 5°C storage at 5 d; yeasts and molds were 8.70 log CFU/g for 25°C storage on the same day. The results of this study determined that S. enterica and L.monocytogenes could survive and grow in dehydrated plant foods during rehydration and storage, highlighting the need for product assessments for these types of foods.
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