• Journal of Microbiology and Biotechnology
• /
• v.32 no.5
• /
• pp.602-611
• /
• 2022

The persistence of pathogenic Escherichia coli under acidic conditions poses a serious risk to food safety, especially in acidic foods such as kimchi. To identify the bacterial factors required for acid resistance, transcriptomic analysis was conducted on an acid-resistant enterotoxigenic E. coli strain and the genes with significant changes in their expression under acidic pH were selected as putative resistance factors against acid stress. These genes included those associated with a glutamate-dependent acid resistance (GDAR) system and copper resistance. E. coli strains lacking GadA, GadB, or YbaST, the components of the GDAR system, exhibited significantly attenuated growth and survival under acidic stress conditions. Accordantly, the inhibition of the GDAR system by 3-mercaptopropionic acid and aminooxyacetic acid abolished bacterial adaptation and survival under acidic conditions, indicating the indispensable role of a GDAR system in acid resistance. Intriguingly, the lack of cueR encoding a transcriptional regulator for copper resistance genes markedly impaired bacterial resistance to acid stress as well as copper. Conversely, the absence of YbaST severely compromised bacterial resistance against copper, suggesting an interplay between acid and copper resistance. These results suggest that a GDAR system can be a promising target for developing control measures to prevent E. coli resistance to acid and copper treatments.

• Genomics & Informatics
• /
• v.19 no.3
• /
• pp.30.1-30.8
• /
• 2021

Salmonella species are among the major pathogens that cause foodborne illness outbreaks. In this study, we aimed to develop a loop-mediated isothermal amplification (LAMP) assay for the rapid and sensitive detection of Salmonella species. We designed LAMP primers targeting the hilA gene as a universal marker of Salmonella species. A total of seven Salmonella species strains and 11 non-Salmonella pathogen strains from eight different genera were used in this study. All Salmonella strains showed positive amplification signals with the Salmonella LAMP assay; however, there was no non-specific amplification signal for the non-Salmonella strains. The detection limit was 100 femtograms (20 copies per reaction), which was ~1,000 times more sensitive than the detection limits of the conventional polymerase chain reaction (PCR) assay (100 pg). The reaction time for a positive amplification signal was less than 20 minutes, which was less than one-third the time taken while using conventional PCR. In conclusion, our Salmonella LAMP assay accurately detected Salmonella species with a higher degree of sensitivity and greater rapidity than the conventional PCR assay, and it may be suitable for point-of-care testing in the field.

• Food Science of Animal Resources
• /
• v.43 no.4
• /
• pp.625-638
• /
• 2023

Among various biological agents, bacteriocins are important candidates to control Listeria monocytogenes which is a foodborne pathogen. In this study, a novel bacteriocin, named agilicin C7, was isolated from Ligilactobacillus agilis C7 showing inhibitory activity against L. monocytogenes. Agilicin C7 biosynthesis gene was characterized by bioinformatics analyses and heterologously expressed in Escherichia coli for further study. The anti-listeria activity of recombinant agilicin C7 (r-agilicin C7) was lost by proteases and α-amylase, suggesting that agilicin C7 is a glycoprotein. r-Agilicin C7 has wide pH and thermal stability and is also stable in various organic solvents. It destroyed L. monocytogenes by damaging the integrity of the cell envelope. These properties of r-agilicin C7 indicate that agilicin C7 is a novel amylase-sensitive anti-listerial Class IId bacteriocin. Physicochemical stability and inhibitory activity against L. monocytogenes of r-agilicin C7 suggest that it can be applied to control L. monocytogenes in the food industry, including dairy and meat products.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.44 no.12
• /
• pp.1895-1904
• /
• 2015

Foodborne illness associated with food service establishments is an important food safety issue in Korea. In this study, foodborne pathogens (Bacillus cereus, Clostridium perfringens, Escherichia coli, pathogenic Escherichia coli, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and Vibrio parahaemolyticus) and hygiene indicator organisms [total viable cell counts (TVC), coliforms] were analyzed for food and environmental samples from foodservice establishments at schools in Gyeonggi province. Virulence factors and antimicrobial resistance of detected foodborne pathogens were also characterized. A total of 179 samples, including food (n=66), utensil (n=68), and environmental samples (n=45), were collected from eight food service establishments at schools in Gyeonggi province. Average contamination levels of TVC for foods (including raw materials) and environmental samples were 4.7 and 4.0 log CFU/g, respectively. Average contamination levels of coliforms were 2.7 and 4.0 log CFU/g for foods and environmental swab samples, respectively. B. cereus contamination was detected in food samples with an average of 2.1 log CFU/g. E. coli was detected only in raw materials, and S. aureus was positive in raw materials as well as environmental swab samples. Other foodborne pathogens were not detected in all samples. The entire B. cereus isolates possessed at least one of the diarrheal toxin genes (hblACD, nheABC, entFM, and cytK enterotoxin gene). However, ces gene encoding emetic toxin was not detected in B. cereus isolates. S. aureus isolates (n=16) contained at least one or more of the tested enterotoxin genes, except for tst gene. For E. coli and S. aureus, 92.7% and 37.5% of the isolates were susceptible against 16 and 19 antimicrobials, respectively. The analyzed microbial hazards could provide useful information for quantitative microbial risk assessment and food safety management system to control foodborne illness outbreaks in food service establishments.

• Journal of Food Hygiene and Safety
• /
• v.26 no.4
• /
• pp.296-301
• /
• 2011

This study was to evaluate the growth potential of E. coli O157:H7 in lettuce leaf extracts and on lettuce leaf surface at various temperatures. The pathogen can survive and multiply in the extracts and leaf surface of lettuce. The population of E. coli O157:H7 in the lettuce extracts reached to 4.79 log CFU/mL at $37^{\circ}C$. The multiplication of pathogen in lettuce extracts initiated within 10 hours of inoculation over $15^{\circ}C$ conditions. And it can survive in the lettuce leaf extracts at $4^{\circ}C$ for 100 hours at least. And this pathogen can multiply on lettuce leaf surface and the population of pathogen on the lettuce leaf surface increased to 1.82 log CFU/g at $25^{\circ}C$. At $37^{\circ}C$, the pathogen density increased to 1.53 CFU/g within 3 days after inoculation. At all temperature, irrespective of the inoculation level, similar trends in growth of E. coli O157:H7 were observed. These results emphasize the growth potential of E. coli O157:H7 in lettuce leaf extract and on lettuce leaf surface. To reduce the risk of outbreak, it is important to maintain the cold chain system during storage before the consumption.

• Journal of Food Hygiene and Safety
• /
• v.34 no.6
• /
• pp.583-590
• /
• 2019

As the consumption of fresh fruits and vegetables increases, food poisoning caused by foodborne pathogen contamination is not decreasing. To prevent the contamination of produce, a quick and easy, low-cost, environmentally-safe disinfection method that does not affect produce freshness or quality is needed. This study demonstrates a new-concept, circulating-water disinfection system that purifies water by using newly developed 'LED-PS (photosensitizer)-induced ROS generation unit'. Using various types of LED-PS induced ROS generation units, we investigated the conditions for reducing the density of various pathogenic bacteria by more than 3 log CFU / mL in 1 hour. The major operational factors affecting the density reduction of the LED-PS-induced ROS generation unit were analyzed. Depending on bacteria species, the density reduction rate was varied. The effect of the units on reducing the density of Bacillus cereus and Pectobacterium carotovorum subsp. carotovorum was high, but the effect on foodborne bacteria such as Escherichia coli was relatively low. In this circulating water disinfection system, the density reduction effect tended to increase as the flow rate increased and the initial bacterial density decreased. As the amount of PS absorbed beads increased, the density reduction effect increased exponentially in some bacteria. Model 3280, a double cylindrical unit connecting two single cylindrical units, could completely sterilize more than 3 log CFU/mL of B. cereus and P. carotovorum subsp. carotovorum in 30 minutes of LED irradiation.

• Journal of Food Hygiene and Safety
• /
• v.39 no.2
• /
• pp.109-117
• /
• 2024

This study investigated the microbial contamination of agricultural, livestock, and marine ingredients in 55 meal kits distributed across Gyeonggi-do, South Korea. Of the 55 meal kits, 48 contained agricultural ingredients, 43 contained livestock ingredients, and 16 contained marine ingredients. The detection rate of the total aerobic bacteria in the agricultural, livestock, and marine products was 100%. The average numbers of the total aerobic bacteria were 6.57 log colony-forming units (CFU)/g in the agricultural products, 4.60 log CFU/g in the livestock products, and 5.47 log CFU/g in the marine products. The coliform detection rates in the agricultural, livestock, and marine products were 81.25%, 69.77%, and 43.75%, respectively. The average numbers of coliforms were 2.83 log CFU/g in the agricultural products, 1.34 log CFU/g in the livestock products, and 1.12 log CFU/g in the marine products. Escherichia coli was detected in 13 livestock products (30.23%), with levels ranging from 0.70 to 2.36 log CFU/g. Contrastingly, E. coli was detected in only one marine product (6.25%) and was not detected in any agricultural products. The detection rates of fungi in agricultural, livestock, and marine products were 97.92%, 93.02%, and 93.75%, respectively. The average numbers of fungi were 3.82 log CFU/g for the agricultural products, 2.92 log CFU/g for the livestock products, and 2.82 log CFU/g for the marine products. The isolation rates of foodborne pathogens from the agricultural, livestock, and marine products were 35.42%, 37.21%, and 31.25%, respectively. Forty-five foodborne pathogens of seven species, including Bacillus cereus and Salmonella spp., were isolated from the raw materials of the agricultural, livestock, and marine products in 55 meal kits. To prevent foodborne diseases caused by meal kits, it is necessary to focus on washing, heating, and preventing cross-contamination during cooking.

• Journal of Applied Biological Chemistry
• /
• v.59 no.1
• /
• pp.63-68
• /
• 2016

This study was performed to assess the microbiological quality of Brassica campestris var. narinosa microgreen from harvesting and processing steps. The samples were analyzed for total viable cell counts (TVC), coliforms, Enterobacteriaceae, Escherichia coli, Salmonella spp., Listeria monocytogenes, Vibrio parahaemolyticus, Bacillus cereus, and Staphylococcus aureus. The total viable counts of microgreen (whole leaves) and environment samples from harvesting steps were higher than 6.8 log CFU/g and the contamination level of coliforms in the samples were 3.2 log CFU/g and 3.5 log CFU/g of microgreen and soil, respectively. In case of microgreen samples collected from processing steps, the contamination level of TVC and coliforms were higher in raw materials than samples obtained from later stages of processing, i.e. washing, drain, and final products. The contamination levels of B. cereus in raw materials and environments decreased approximately 1.4 log CFU/g in final products. S. aureus was detected in soil samples but Salmonella spp., Listeria monocytogenes, Vibrio parahaemolyticus and pathogenic E. coli was not detected. In order to identify the sources of contamination for microgreen, the genetic similarity of B. cereus isolates obtained from harvesting and processing steps were compared using the repetitive-sequence-based polymerase chain reaction method. B. cereus isolates obtained from harvesting environments and microgreen were clustered with a similarity greater than 95%. In case of B. cereus isolates obtained from microgreen and environmental samples at processing steps showed low genetic similarity.

• Food Science and Preservation
• /
• v.21 no.3
• /
• pp.427-433
• /
• 2014

Ethanol and hot water extracts were prepared from Lindera obtusiloba Blume (LO) and Zanthoxylum piperitum DC (ZP) and used to evaluate their antimicrobial activities and thermal stability against six foodborne pathogens (3 gram-positive and 3 gram-negative bacteria). The antimicrobial activities were assessed using the agar diffusion method, and the thermal stabilities of extracts were examined after heat treatment at 60, 70, 80, and $100^{\circ}C$ for 10 min. The zones of inhibition by the LO extract or the ZP extract of the tested microorganisms were in the range of 21-30 mm and 19-25 mm, respectively, at 100 mg/mL concentrations. The 60% ethanol extract and the hot water extracts from LO showed the strongest antimicrobial effects against MRSA and Staphylococcus aurues, respectively. For the extract from ZP, the strongest antimicrobial effect was shown against S. aurues by 60% ethanol, and the weakest antimicrobial effect was shown against E. coli by the hot water extracts. The ZP extracts showed that the gram-positive bacteria were more sensitive than gram-negative bacteria. For the thermal stability of the extracts, the antimicrobial effects stabilized after heat treatment. Overall, the data suggest that the extracts have a potential for application in various food products for which a natural antimicrobial additive is desired.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.38 no.9
• /
• pp.1265-1270
• /
• 2009

Pathogens contaminated on the surface of utensils could contribute to the occurrence of outbreaks due to cross-contamination to foods during the food preparation process. Therefore, the efficacy of chemical sanitizers (chlorine-based and alcohol-based commercial sanitizers) and dry-heat ($71^{\circ}C$) on inhibiting biofilms of five foodborne pathogens (Escherichia coli O157:H7, Salmonella Typhimurium, Pseudomonas aeruginosa, Listeria monocytogenes, and Staphylococcus aureus) on the surface of stainless steel and polypropylene were investigated in this study. Initial populations of pathogens were 8.8$\sim$9.3 and 9.4$\sim$10.3 log CFU/coupon on the surface of stainless steel and polypropylene coupon, respectively, and these populations were not significantly reduced when they were treated with water for 5 min at room temperature. Treatments with chlorine sanitizer and dry-heat were not effective on inactivating pathogens on the surfaces of stainless steel and polypropylene. In contrast, treatments with alcohol sanitizer were very effective on inactivating pathogens on the surfaced of stainless steel and polypropylene. Reduction levels ranged from 3.4 to 6.4 log and from 5.5 to 7.4 log CFU/coupon in stainless steel and plastic coupons, respectively. From these results, alcohol-based sanitizer could be used as a potential way for controlling microbial contamination on the surface of utensils, cooking equipment, and other related environments.