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Acid-stress adaptation in Listeria monocytogenes (Lm) serotype 4b and 1/2a occurred when cells were pre-exposed to pH 5.0 tryptic soy broth supplemented with yeast extract (TSB-YE) at 22°C or 37°C but not at 4°C. Prolonged time, varied sublethal acid pH, substitute of acidulants and addition of sodium chloride during 4°C mild acid pre-exposure still did not induce acid-stress adaptation in Lm. This finding was also validated using an acidic cheese, similar to what has seen for Gram-negative bacteria E. coli and Salmonella. Further investigation revealed that major cold shock protein in Lm CspL was not responsible for repressed acid-stress adaptation at 4°C. A bead beating treatment prior to mild acid pre-exposure at 4°C partially induced acid-stress adaptation after pre-exposure in 4°C to mild acid stress. Our data suggests that cold processing or cold storage temperature can lower the possibility of activating acid-stress adaptation in Lm.

Previous findings showed that Listeria monocytogenes exhibits higher heat tolerance to thermal exposure at 60°C when pre-exposed to sublethal heat stress at 48°C for 30/60 min. The objective of this study was to investigate the morphological changes that occurred in L. monocytogenes serotype 1/2a cells as visualized by transmission electron microscopy (TEM) after exposure to sublethal heat stress at 48°C for 60 min and in combination with lethal concentration of carvacrol for 30 min. The TEM micrographs revealed thickening of cell wall and cell membrane, and clumping of cytoplasm when subjected to sublethal heat stress followed by carvacrol treatment. These studies indicate that L. monocytogenes cells when adapted to heat stress may alter its morphology to protect themselves against carvacrol. Further studies will investigate the effect of increased contact time with carvacrol and with other essential oils on the ultrastructural changes in L. monocytogenes cells adapted to heat stress.

The objective of this study was to investigate the differences in survival of L. monocytogenes EGD (Bug600) (serotype 1/2a) in various disinfectants and essential oils after sublethal heat stress at 48°C for 60 min. The survival of heat stressed L. monocytogenes cells was decreased in lethal acid (HCl or H3PO4 pH 2.5), lethal oxidative stress (H2O2 1000 ppm) and lethal quaternary ammonium compounds (QAC 2.5-3.5 ppm) compared to non-stressed control cells. By contrast, the survival of heat stressed L. monocytogenes cells was higher in lethal alkali (NaOH or KOH pH 12) than the control cells. Also, the survival of heat stressed L. monocytogenes cells was higher in lethal carvacrol (428 ppm) and bay oil (1100 ppm) but was decreased in lethal red thyme oil (300 pm) compared to control. These findings indicate that the heat stressed cells of L. monocytogenes are not easily killed by alkali-based disinfectants and essential oils containing carvacrol and bay oil. Therefore, disinfectants and essential treatments should be carefully considered when heat stressed cells of L. monocytogenes may be present.

According to USDA-FSIS reports, L. monocytogenes and Salmonella are two important foodborne pathogens that are prevalent throughout catfish environment. Channel catfish (Ictalurus punctatus) is the most important aquaculture species in the United States accounting for more than 60% of its aquaculture production. The objectives of this study were to determine the effect of strain, temperature concentration on the growth, survival and biofilm formation of L. monocytogenes and Salmonella using catfish mucus extract on different food-contact surfaces. Growth and survival of L. monocytogenes and Salmonella was greater at the higher concentration of mucus extract at both 10°C and 22°C. In 15 micrograms/ml catfish mucus extract L. monocytogenes and Salmonella counts increased to 4.5 log CFU/ml after 72 h at 10°C and 5-7 log CFU/ml counts after 32-48 h at 22°C. In 375 micrograms/ml catfish mucus extract L. monocytogenes and Salmonella counts increased to 6-7 log CFU/ml counts after 72 h at 10°C and 8-9 log CFU/ml counts after 32-48 h at 22°C. L. monocytogenes and Salmonella were able to grow and survive for more than 63 days with at least 4-6 log CFU/ml at 10°C and 6-8 log CFU/ml in 375 micrograms/ml and 15 micrograms/ml of catfish mucus extract respectively. No differences (P > 0.05) among L. monocytogenes and Salmonella strains were observed to form biofilms in the presence catfish mucus extract on the stainless steel surface. The biofilm formation by L. monocytogenes and Salmonella in catfish mucus extract was less (P < 0.05) on buna-n rubber when compared to stainless steel, polyethylene and polyurethane surfaces. Therefore, the findings in this study show that catfish mucus promotes L. monocytogenes and Salmonella to grow and subsequently form biofilms on different food-contact surfaces also promotes growth and survival of these pathogens for longer period of time in catfish processing industries.

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Listeria monocytogenes is a major concern for the food industry. It is one of the major agents causing listeriosis. The objective of the first part of this study is to evaluate the effect of hydration level on attached listeria monocytogenes on stainless steel/High density polyethylene transferred to food products. Attached cells were prepared on stainless steel/High density polyethylene. Transfer experiments were conducted from inoculated surface material slides to cold smoked salmon fillets. This experiment was repeated 6 times. The results were analyzed with an analysis of variance by SAS. The differences between the different RH% and surface conditions were not statistically significant. There was variability in between packages, brands and over the course of storage after opening, and likely contributed to the variability of transfer observed in this set of experiments. The objective of the second part of the research is to study the effect of hydration level on the detachment of Listeria monocytogenes biofilm growing on stainless steel by using Atomic force microscope. Biofilms were grown on stainless steel in drip flow bioreactor at 32 °C for 72 h. Then biofilms were equilibrated over saturated salt solution at 20 °C for 48 h before the Atomic Force Microscope experiment. The results showed that cohesive energy value of the biofilm increased with biofilm depth. Only square shaped displaced 2.5X2.5 μm region were visualized after serious of raster scanning under high load which means that moisture condition of Listeria monocytogenes biofilm can significantly affect the cohesiveness between of Listeria monocytogenes biofilm.

The food-borne pathogen Listeria monocytogenes can attach to the environmental surfaces and develop biofilm which can cause food contamination in the food industries. Sortase A and surface proteins are involved in biofilm and virulence of L. monocytogenes. Curcumin was reported to inhibit sortase A and biofilm in gram positive bacteria. The overall objective of this study was to observe the effect of curcumin (Curcuma longa) on the biofilm formation and surface proteins of L. monocytogenes. The antibiofilm effect of curcumin against the strain LM21 (wild type) and s22-11G (sortase A defective mutant) was studied using the microtiter plate assay. No significant differences between the growth of the wild type and the sortase A defective mutant were observed at sub-inhibitory concentrations of curcumin. However, a greater biofilm reduction was observed in the strain s22-11G. The effect of curcumin from two different manufacturers on the wild type was also compared by the microtiter plate assay. Both curcumin did not exhibit statistically different effect on the growth of the wild type. However, a greater biofilm inhibitory effect was observed in one curcumin. The HPLC results suggested that curcumin with the greater antibiofilm activity contained higher amount of curcumin which was reported to be the most potent curcuminoid compound in curcumin. Three different protein extraction methods were evaluated and the most efficient method was used for 2D-GE. When cells were grown in the presence of curcumin, 5 proteins, 16 proteins and 4 proteins were up-regulated, down-regulated and absent, respectively in L. monocytogenes LM21. The influence of the enzyme sortase A upon surface protein expression was evaluated by comparing proteins expressed by wildtype L. monocytogenes LM21 to that of the sortase A mutant, s22-11G. In strain s22-11G, 2 proteins, 8 proteins and 3 proteins were up-regulated, down-regulated and absent in comparison to wildype LM21. The exact information of these differentially expressed proteins still need to be identified by mass spectrometry.

Listeria monocytogenes is an important foodborne pathogen that can adapt to stress conditions to persist in food processing environments. Our findings show that there was a development of low-level tolerance to quaternary ammonium compound (QAC) and antibiotics ciprofloxacin and trimethoprim in L. monocytogenes after sublethal adaptation to QAC. Using eight L. monocytogenes strains, we determined the changes in MIC, growth rate, and surviving CFU for homologous and heterologous stress-response after sublethal exposure to daily cycles of fixed or gradually increasing concentration of QAC. Three main findings were observed: (1) Short-range MIC of QAC, ciprofloxacin, and trimethoprim increased by 1.6 to 2.3, 1.5 to 2.9, and 1.7 to 2.5 fold against QAC-adapted phenotypes of L. monocytogenes as compared to the non-adapted cells; (2) QAC-adapted phenotypes of L. monocytogenes exhibited a significant increase in growth rate by 2.5 to 7.1, 2.1 to 6.8, or 1.4 to 4.8 fold in the broth model containing QAC, ciprofloxacin, or trimethoprim respectively, as compared to non-adapted cells; and (3) QAC-adapted phenotypes of L. monocytogenes exhibited a significant increase in survival by 1.5 to 4, 2.2 to 4.3, or 1.3 to 3.2 log CFU/ml in the agar model containing QAC, ciprofloxacin, or trimethoprim respectively, as compared to non-adapted cells (P < 0.05). There were strain differences in QAC-adapted phenotypes of L. monocytogenes for both homologous and heterologous stress-response with some strains exhibiting a significant increase in short-range MIC, growth rate, and survival while others exhibiting no changes as compared to non-adapted cells. These findings suggest the potential formation of low-level QAC-tolerant and antibiotic-tolerant phenotypes in some L. monocytogenes strains under residual QAC concentrations (where QAC may be used widely) and such cells if not inactivated may survive longer to increase food safety risk.

No description available.