• Journal of Microbiology
• /
• v.45 no.1
• /
• pp.48-52
• /
• 2007

A new procedure for the concentration of nonoviruses from water samples has been developed. This procedure (calcium flocculation-citrate dissolution method) uses the following steps: virus flocculation formed by treatment with 1 M $CaCl_2$ and 1 M $Na_2HPO_4$, virus release by sodium citrate dissolution (0.3 M Na citrate, pH 3.5), and virus re-concentration by ultrafiltration. When reverse transcription (RT)-PCR was performed after the procedure, the overall detection sensitivity for seeded noroviruses in a one liter drinking water sample was as low as 1 RT-PCR unit, which is equal to a $10^{-6}$ dilution of the virus sample. This approach showed at least a 5-fold-higher sensitivity than the current method with its three steps of adsorption-elution-concentration. The newly developed procedure was used to test different brands of bottled drinking water from China for putative contamination with noroviruses. A total of 144 samples were analyzed; all of the samples were negative for norovirus specific nucleic acids.

• Food Science and Biotechnology
• /
• v.18 no.3
• /
• pp.788-792
• /
• 2009

Gamma $({\gamma})-irradiation$ can be used to control pathogens such as Vibrio vulnificus in seafood. The effects of irradiation on microbial cell populations (%) have been studied in order to develop detection methods for irradiated foods. The method used in this study was ethidium bromide monoazide (EMA) real-time polymerase chain reaction (PCR), using V. vulnificus specific primer, EMA, and $SYBR^{(R)}$ Green to discriminate between ${\gamma}-irradiated$ and non-irradiated cells. Confocal microscope examination showed that ${\gamma}-irradiation$ damaged portions of the cell membrane, allowing EMA to penetrate cells of irradidated V. vulnificus. ${\gamma}-Irradiation$ at 1.08 KGy resulted in log reduction ($-1.15{\pm}0.13$ log reduction) in genomic targets derived from EMA real-time PCR. The combination cold/heat shock resulted in the highest ($-1.74{\pm}0.1$ log reduction) discrimination of dead irradiated V. vulnificus by EMA real-time PCR.

• Food Science and Biotechnology
• /
• v.14 no.3
• /
• pp.410-412
• /
• 2005

Vibrio parahaemolyticus is a gram-negative bacterium which acts as a causative agent for food poisoning. Studies with respect to specific extracellular proteins of V. parahaemolyticus would be useful for the development of specific detection methods against V. parahaemolyticus. In our present study, outer membrane proteins (OMPs) of V. parahaemolyticus were obtained from insoluble traction of 1% sarkosyl treated-cell wall materials. SDS-PAGE analysis showed the presence of several conserved outer membrane proteins among five strains of V. parahaemolyticus, and three bands were identified as V. parahaemolyticus OMPs through MALDI-TOF analysis. Polyclonal antibodies enriched with anti-OmpU were obtained from immunized rabbits. The antibodies against these proteins may be useful for the development of detection methods for V. parahaemolyticus.

• Asian-Australasian Journal of Animal Sciences
• /
• v.29 no.8
• /
• pp.1188-1196
• /
• 2016

This study evaluated the risk of Clostridium perfringens (C. perfringens) foodborne illness from natural and processed cheeses. Microbial risk assessment in this study was conducted according to four steps: hazard identification, hazard characterization, exposure assessment, and risk characterization. The hazard identification of C. perfringens on cheese was identified through literature, and dose response models were utilized for hazard characterization of the pathogen. For exposure assessment, the prevalence of C. perfringens, storage temperatures, storage time, and annual amounts of cheese consumption were surveyed. Eventually, a simulation model was developed using the collected data and the simulation result was used to estimate the probability of C. perfringens foodborne illness by cheese consumption with @RISK. C. perfringens was determined to be low risk on cheese based on hazard identification, and the exponential model ($r=1.82{\times}10^{-11}$) was deemed appropriate for hazard characterization. Annual amounts of natural and processed cheese consumption were $12.40{\pm}19.43g$ and $19.46{\pm}14.39g$, respectively. Since the contamination levels of C. perfringens on natural (0.30 Log CFU/g) and processed cheeses (0.45 Log CFU/g) were below the detection limit, the initial contamination levels of natural and processed cheeses were estimated by beta distribution (${\alpha}1=1$, ${\alpha}2=91$; ${\alpha}1=1$, ${\alpha}2=309$)${\times}$uniform distribution (a = 0, b = 2; a = 0, b = 2.8) to be -2.35 and -2.73 Log CFU/g, respectively. Moreover, no growth of C. perfringens was observed for exposure assessment to simulated conditions of distribution and storage. These data were used for risk characterization by a simulation model, and the mean values of the probability of C. perfringens foodborne illness by cheese consumption per person per day for natural and processed cheeses were $9.57{\times}10^{-14}$ and $3.58{\times}10^{-14}$, respectively. These results indicate that probability of C. perfringens foodborne illness by consumption cheese is low, and it can be used to establish microbial criteria for C. perfringens on natural and processed cheeses.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.32 no.7
• /
• pp.991-996
• /
• 2003

Determination of regional microbial contamination in the process of rice rolled in dried laver (Kimbab) and effects of gamma irradiation on the improvement of hygienic quality and shelf stability were investigated. Total aerobic bacterial distribution of raw materials of Kimbab were; 10$^{6}$ ∼10$^{7}$ CFU/g in dried laver, 10$^3$ CFU/g in cucumber and below 10 CFU/g in steamed rice, ham, fried egg, and salted radish. Total coliform bacteria were 10$^3$ CFU/g in dried laver and detected below detection limit (10 CFU/g) in other raw materials. And it was arithmetically calculated that the levels of total aerobic bacteria and coliform bacteria in Kimbab does not exceed 10$^{5}$ CFU/g and 10$^1$ CFU/g under the aseptic process, respectively. However, microbial contamination levels in just prepared Kimbab in a market were about 10$^{6}$ CFU/g of total aerobic and coliform bacteria. Therefore, it was considered that microbial contamination of Kimbab is mainly originated from environmental uptake during the preparation. The representative media for putrefying bacterial growth were steamed rice. Coliform bacteria were mainly increased in ham and fried egg during storage. The bacteria in dried laver were radio-resistant and survived at 3 kGy of gamma irradiation. Coliform bacteria on EMB agar plate were eliminated at the dose of 2 kGy. The sensory acceptability of 2 kGy irradiated Kimbab was stable and the Kimbab can be preserved for 24 hour at 15$^{\circ}C$. Therefore, it was considered that optimal irradiation dose for radicidation of Kimbab was 2 kGy.

• Journal of the Society of Cosmetic Scientists of Korea
• /
• v.40 no.3
• /
• pp.307-311
• /
• 2014

2,3,5-Triphenyltetrazolium chloride (TTC) is used as a redox indicator in culture media. It is colorless in the oxidized form and is reduced to formazan, an insoluble pigment, by dehydrogenases in actively growing microbial cells. The aim of this study was to assess by microbial test of the pathogenic microorganisms using TTC reduction. The pathogenic microorganisms were reduced in medium by dehydrogenase to produce insoluble red formazan. We observed that the optimization method of TTC allowed more than 12 h incubation in 0.04% concentration. Also, the growth of microorganisms with media was increased formazan production. We confirmed that microorganisms were quickly observed to grow colonies cultured red color without affecting the growth of microorganisms. It is suggested that the microbial test using TTC can provide better and quicker test method in cosmetics development.

• Journal of Food Hygiene and Safety
• /
• v.26 no.2
• /
• pp.182-185
• /
• 2011

This study was conducted to develop an appropriated management for safety of children snacks sold around school. Total 598 items as targeted food were collected; 66 biscuits, 320 candies, 57 chocolates, 40 ice creams and 115 beverages. Microbiological hazards such as total aerobic bacteria, Coliforms, Escherichia coli, Bacillus ceruse, Yeasts & molds were measured by analytical method in Korean food code. Total aerobic bacteria and Yeasts & molds were detected in cookies at the level of less than 2.69 and 2.65 $log_{10}$ CFU/g and the detection rates were 54.55 and 62.12%, respectively. Bacillus cereus was detected in 1 snack only at the level of 1.39 $log_{10}$ CFU/g but it was less than Korean microbial standards and specifications (3 $log_{10}$ CFU/g). Total aerobic bacteria and Yeasts & molds were detected in candies less than 2.86, 3.36 $log_{10}$ CFU/g and the detection rates were 46,8% respectively. Total aerobic bacteria, Yeast & mold were detected in chocolates at the levels less than 2.52 and 1.87 $log_{10}$ CFU/g and the detection rates were 33 and 22% respectively. Total aerobic bacteria in both ice creams and beverages were detected at the levels less than 3.39 and 1.35 $log_{10}$ CFU/g and the detection rates were 82 and 5% respectively. Coliforms were found in one ice cream (1.39 $log_{10}$ CFU/g) only. The result of this study indicated that all children snacks around school were suitable for microbial standard and specifications in Korean Food Code. However, since most children snacks around school are circulated without proper storage temperature and handing condition, consistent microbial management for children snacks are needed.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.36 no.4
• /
• pp.333-339
• /
• 2016

The objective of this study was to optimize analytical methods for ochratoxin A (OTA) and zearalenone (ZEA) in rice straw silage and winter forage crops using ultra-high performance liquid chromatography (UHPLC). Samples free of mycotoxins were spiked with $50{\mu}g/kg$, $250{\mu}g/kg$, or $500{\mu}g/kg$ of OTA and $300{\mu}g/kg$, $1500{\mu}g/kg$, or $3000{\mu}g/kg$ of ZEA. OTA and ZEA were extracted by acetonitrile and cleaned-up using an immunoaffinity column. They were then subjected to analysis with UHPLC equipped with a fluorescence detector. The correlation coefficients of calibration curves showed high linearity ($R^2{\geq_-}0.9999$ for OTA and $R^2{\geq_-}0.9995$ for ZEA). The limit of detection and quantification were $0.1{\mu}g/kg$ and $0.3{\mu}g/kg$, respectively, for OTA and $5{\mu}g/kg$ and $16.7{\mu}g/kg$, respectively, for ZEA. The recovery and relative standard deviation (RSD) of OTA were as follows: rice straw = 84.23~95.33%, 2.59~4.77%; Italian ryegrass = 79.02~95%, 0.86~5.83%; barley = 74.93~97%, 0.85~9.19%; rye = 77.99~96.67%, 0.33~6.26%. The recovery and RSD of ZEA were: rice straw = 109.6~114.22%, 0.67~7.15%; Italian ryegrass = 98.01~109.44%, 1.65~4.81%; barley = 98~113.53%, 0.25~5.85%; rye = 90.44~108.56%, 2.5~4.66%. They both satisfied the standards of European Commission criteria (EC 401-2006) for quantitative analysis. These results showed that the optimized methods could be used for mycotoxin analysis of forages.

• Korean journal of food and cookery science
• /
• v.31 no.1
• /
• pp.1-8
• /
• 2015

Organic acids are known as natural sanitizers. We examined the sanitizing effects of five organic acids (acetic acid, propionic acid, citric acid, malic acid, and lactic acid) and their persistence on three pectinolytic yeast strains isolated from decayed citrus, and the persistence of their sanitizing effects was determined during storage at $4^{\circ}C$ and $16^{\circ}C$. The 7~8 log CFU/mL of the mixed three yeast mixture was exposed to various concentrations of each organic acid for 1 min. The yeast mixtures decreased under detection limit(1 log CFU/mL) in 1% of acetic acid, followed by in 3% of propionic acid with the reduction of 5 log CFU/mL. The citric acid, malic acid, and lactic acid decreased the number of yeasts under detection limit at 7.5%. When treated with deionized water and 1~5% of organic acids were treated on the surfaces of citrus contaminated by yeasts, total numbers of the yeasts decreased under detection limit(3 log CFU) at 5% of acetic acid and 4 log CFU/piece at 5% propionic acid compared with deionized water. When treated with acetic acid and propionic acid on the stem ends of the contaminated citrus, total numbers of the yeasts significantly decreased 0.5 log CFU/piece at 3% of both organic acids. During storage at $4^{\circ}C$ and $16^{\circ}C$ for 20 days, total number of yeasts significantly decreased at 2% acetic acid compared with deionized water. This study suggested that organic acids could be used to sanitize microbial contaminants from citrus for storage and transportation.

• Korean Journal of Environmental Biology
• /
• v.20 no.3
• /
• pp.189-196
• /
• 2002

The direct detection of intestinal pathogens and viruses often requires costly, tedious, and time-consuming procedures. These requirements developed a test to show that the water was contaminated with sewage-borne pathogens by assessing the hygienic quality of water based on indicator microorganisms whose presence indicates that pathogenic microorganisms may also be present. Various groups of microorganisms have been suggested and used as indicator microorganisms. Proposed and commonly used microbial indicators are total coliforms, fecal coliforms, fecal streptococci, Clostridium perfringens, heterotrophic plate count, bacteriophage, and so on. Unfortunately, most, if not all, of these indicators are not ideal because of the sensitivity and resistance to environment stresses and disinfection. However, the development of gene probes and PCR technology may give hope for the discovery of rapid and simple methods toy detecting small number of fecal pathogens in various environments.