The development and commercialization of industrial genetically modified (GM) organisms is actively progressing worldwide, highlighting an increased need for improved safety management protocols. We sought to establish an environmental monitoring method, using real-time polymerase chain reaction (PCR) and propidium monoazide (PMA) treatment to develop a quantitative detection protocol for living GM microorganisms. We developed a duplex TaqMan quantitative PCR (qPCR) assay to simultaneously detect the selectable antibiotic gene, ampicillin (AmpR), and the single-copy Escherichia coli taxon-specific gene, D-1-deoxyxylulose 5-phosphate synthase (dxs), using a direct cell suspension culture. We identified viable engineered E. coli cells by performing qPCR on PMA-treated cells. The theoretical cell density (true copy numbers) calculated from mean quantification cycle (Cq) values of PMA-qPCR showed a bias of 7.71% from the colony-forming unit (CFU), which was within ±25% of the acceptance criteria of the European Network of GMO Laboratories (ENGL). PMA-qPCR to detect AmpR and dxs was highly sensitive and was able to detect target genes from a 10,000-fold (10-4) diluted cell suspension, with a limit of detection at 95% confidence (LOD95%) of 134 viable E. coli cells. Compared to DNA-based qPCR methods, the cell suspension direct PMA-qPCR analysis provides reliable results and is a quick and accurate method to monitor living GM E. coli cells that can potentially be released into the environment.
Journal of the Korean Society of Food Science and Nutrition
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제34권1호
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pp.66-74
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2005
This survey was conducted to assess the consumer perceptions of food-related hazard in 500 housewives from all over Korea. The subjects were selected by stratified random sampling method. The survey was performed using structured questionnaire through telephone interview by skilled interviewers. The results showed that 34.6% of the respondents felt secure and were not concerned about food safety, and 65.4% were concerned about food safety. Logistic regression analysis showed that the increasing concern on food brands, food additives (such as food preservatives and artificial color), and imported foodstuffs indicated the current increasing concern on food safety. Other related factors indicating the increasing concern on food safety were education level and care for children's health. The respondents who cared about food safety expressed a high degree of concern on processed foodstuffs such as commercial boxed lunch (93.3%), imported foods (92.7%), fastfoods (89.9%), processed meat products (88.7%), dining out (85.6%), cannery and frozen foods (83.5%), and instant foods (82.0%). The lowest degree of concern was on rice. All the respondents perceived that residues of chemical substances such as pesticides and food additives, and endocrine disrupters were the most potential food risk factors, followed by food-borne pathogens, and GMOs (Genetically Modified Organisms). However, these results were not consistent with scientific judgment. Therefore, more education and information were needed for consumers' awareness of facts and myths about food safety. In addition, the results showed that consumers put lower trust in food products information such as food labels, cultivation methods (organic or not), quality labels, and the place of origin. Nevertheless, the respondents expressed their desire to overcome alienation, and recognized the importance of knowing of the origin or the producers of food. They identified that people who need to take extreme precautions on food contamination were the producers, government officials, food companies, consumers, the consumer's association, and marketers, arranged in the order of highest to lowest. They also believed that the production stage of agriculture was the most important step for improving the level of food safety Therefore, the results indicated that there is a need to introduce safety systems in the production of agricultural products, as follows: Good Agricultural Practice (GAP), Hazard Analysis and Critical Control Point (HACCP), and Traceability System (75).
Woo, So Young;Lee, Sang Yoo;Tian, Fei;Jeong, A-Yeong;Yoo, Cha Nee;Kang, Seung Yoon;Chun, Hyang Sook
Journal of Food Hygiene and Safety
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제35권5호
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pp.438-446
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2020
Meju and nuruk (respectively soybean and malt) are traditional Korean fermentation starters that are vulnerable to contamination by harmful microorganisms such as aflatoxigenic fungi and their associated aflatoxins (AFs). In this study, Aspergillus spp. were isolated and identified from a total of 57 meju and 18 nuruk samples collected from Korean markets. Their potential aflatoxigenicity was investigated by examining the presence of three aflatoxin biosynthetic genes (aflO, aflP, and aflR) using multiplex polymerase chain reaction (mPCR) assays. Thereafter, aflatoxin production of isolates and the natural occurrence of AFs in meju and nuruk samples were analyzed by high-performance liquid chromatography (HPLC). A total of 177 Aspergillus isolates were identified and 130 isolates were obtained from meju samples. Of these, 25 isolates (19.2%) contained all three aflatoxin biosynthetic genes, and five (20%) of these isolates produced aflatoxins. Forty-seven of the Aspergillus isolates were obtained from nuruk samples, five of which (10.6%) expressed all three AF biosynthetic genes; however, none of these strains produced AFs. HPLC analysis showed that 88% (51/58) of the meju samples and 39% (7/18) of nuruk samples were not contaminated with AFs (below limit of detection). Among the isolates isolated from meju and nuruk, there were aflatoxigenic strains containing all three aflatoxin biosynthetic genes or producing aflatoxin in medium, but the frequency of aflatoxin contamination was low in the meju and nuruk samples.
Fusarium head blight is an important disease of small grains. It is mainly caused by members of the Fusarium graminearum species complex (FGSC). Barley and wheat growers spray fungicides, especially demethylation-inhibitor fungicides, to suppress the disease. The objective of this study was to examine the changes in the sensitivity of the FGSC population to the triazole fungicide, propiconazole. A total of 124 and 350 isolates of FGSC were obtained from barley and wheat in Jeolla Province during 2010-2016 and 2020-2021, respectively. The species identity and trichothecene chemotypes of the FGSC isolates were determined based on polymerase chain reaction assays targeting translation elongation factor 1-alpha and TRI12 genes, respectively. Sensitivity to propiconazole was determined based on the effective concentration that reduced 50% of the mycelial growth (EC50) using the agar dilution method. Of all isolates, F. asiaticum with the nivalenol chemotype was the most common (83.9% in 2010-2016 and 96.0% in 2020-2021), followed by F. asiaticum with the 3-acetyl deoxynivalenol chemotype (12.1% in 2010-2016 and 2.9% in 2020-2021). The EC50 values of the isolates collected in 2010-2016 and 2020-2021 ranged from 0.0180 to 11.0166 ㎍/mL and 1.3104 to 17.9587 ㎍/mL, respectively. The mean EC50 value of the isolates increased from 3.8648 ㎍/mL in 2010-2016 to 5.9635 ㎍/mL in 2020-2021. The baseline resistance to propiconazole was determined to be 7 ㎍/mL, based on the EC50 value of isolates collected in 2010-2016, and the ratio of resistant isolates increased from 9.7% in 2010-2016 to 28.6% in 2020-2021.
Objective: The aim of this study was to investigate the reversibility and safety of KISS1 metastasis suppressor (KISS1) gene vaccine in immunocastration. Methods: Six eight-week old ram lambs were randomly divided into vaccinated and control groups. The vaccine (1 mg/ram lamb) was injected at weeks 0, 3, and 6 of the study. Blood samples were collected from the jugular vein before primary immunization and at weeks 2, 4, 6, 10, 14, 22, and 30 after primary immunization. All ram lambs were slaughtered at 38 weeks of age, and samples were collected. Results: The specific anti-KISS1 antibody titers in vaccinated animals were significantly higher and the serum testosterone level was significantly lower than those in the control groups from week 4 to 14 after primary immunization (p<0.05). No significant difference was observed at weeks 22 and 30 after the primary immunization. Similar results were also found for scrotal circumference, testicular weight, length, breadth, and spermatogenesis in seminiferous tubules in week 30 after primary immunization. KS (KISS1-hepatitis B surface antigen S) fusion fragment of KISS1 gene vaccine was not detected in host cell genomic DNA of 9 tissues of the vaccinated ram lambs by polymerase chain reaction. Conclusion: The effects of KISS1 gene vaccine in immunocastration were reversible and no integration events were recorded.
A few authors have already investigated microbial contamination of leafy vegetables in distribution condition and examined the effect of temperature abuse on microbial safety or product quality. But this study analyzed proliferation of indicator microorganisms and food poisoning bacteria in real situation in Korean agroindustry and investigated washing effect of micobial contamination on leaf surface. Leafy vegetables were found to be contaminated with aerobic bacteria at $0.1{\sim}0.32{\times}10^3CFU/g$. Staphylococcus aureus and Bacillus cereus were detected in leafy vegetables at $0{\sim}0.54{\times}10^3CFU/g$. However, Salmonella spp., Coliforms, Clostridium spp. and Listeria spp. were not detected in any samples. Indicator microorganisms and food poisoning bacteria on leafy vegetables increased at room temperature when simulatively distributed condition. After clean-up, the indicator microorganisms and food poisoning bacteria of kimchi cabbage, lettuce and perilla leaf were decreased to a very low level. This study suggests that it is necessary to improve the level of hygiene management such as use of cold chain system and hygiene management of transport tools during the distribution process for fresh leafy vegetables.
In, Byung-Chun;Kim, Ji-Gang;Nimikeatkai, Hataitip;Lee, Jung-Soo
Journal of Bio-Environment Control
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제19권4호
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pp.343-350
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2010
This study was conducted to investigate the effect of harvest seasons on quality and microbial population at different steps of production chain of fresh-cut iceberg lettuce. Iceberg lettuces harvested in May, June, July, October, and December were processed following industrial practices, and stored at $5^{\circ}C$ for 9 days. For microbial measurement, samples were taken from each of the following steps: harvest, transport, pretreatment, cutting, 1st-washing, 2nd-washing, and day 3, 6, and 9 of storage. Iceberg lettuce cultivated in protect house and harvested in May and October showed higher $CO_2$ levels in the packages and electrolyte leakages than lettuce harvested in June, July and December. Microbial population of raw materials harvested in July was highest (6.76 log), and microbial growth rate during storage was highest in samples harvested in May. Lettuce harvested in June had better quality and microbial safety compared to other lettuces. Although lettuce harvested in October and December had less microbial population in either raw materials or processed products, those samples had inferior quality due to off-odor development and severe browning. Therefore, it is required to maintain quality and ensure microbial safety to distribute fresh-cut lettuce with high quality and safety throughout the year.
Background: Monitoring and control of PM2.5 are being recognized as key to address health issues attributed to PM2.5. Availability of low-cost PM2.5 sensors made it possible to introduce a number of portable PM2.5 monitors based on light scattering to the consumer market at an affordable price. Accuracy of light scatteringe-based PM2.5 monitors significantly depends on the method of calibration. Static calibration curve is used as the most popular calibration method for low-cost PM2.5 sensors particularly because of ease of application. Drawback in this approach is, however, the lack of accuracy. Methods: This study discussed the calibration of a low-cost PM2.5-monitoring device (PMD) to improve the accuracy and reliability for practical use. The proposed method is based on construction of the PM2.5 sensor network using Message Queuing Telemetry Transport (MQTT) protocol and web query of reference measurement data available at government-authorized PM monitoring station (GAMS) in the republic of Korea. Four machine learning (ML) algorithms such as support vector machine, k-nearest neighbors, random forest, and extreme gradient boosting were used as regression models to calibrate the PMD measurements of PM2.5. Performance of each ML algorithm was evaluated using stratified K-fold cross-validation, and a linear regression model was used as a reference. Results: Based on the performance of ML algorithms used, regression of the output of the PMD to PM2.5 concentrations data available from the GAMS through web query was effective. The extreme gradient boosting algorithm showed the best performance with a mean coefficient of determination (R2) of 0.78 and standard error of 5.0 ㎍/㎥, corresponding to 8% increase in R2 and 12% decrease in root mean square error in comparison with the linear regression model. Minimum 100 hours of calibration period was found required to calibrate the PMD to its full capacity. Calibration method proposed poses a limitation on the location of the PMD being in the vicinity of the GAMS. As the number of the PMD participating in the sensor network increases, however, calibrated PMDs can be used as reference devices to nearby PMDs that require calibration, forming a calibration chain through MQTT protocol. Conclusions: Calibration of a low-cost PMD, which is based on construction of PM2.5 sensor network using MQTT protocol and web query of reference measurement data available at a GAMS, significantly improves the accuracy and reliability of a PMD, thereby making practical use of the low-cost PMD possible.
Kim, Byoung-Sik;Kim, Min;Heo, Kwang-Beom;Hong, Joo-Hee;Na, Won-Jae;Kim, Jae-Hun
Applied Chemistry for Engineering
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제18권1호
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pp.10-16
/
2007
In this study, we examined the preparation and hydrolysis property of immobilized urease membrane to decompose harmful urea in the body and remove ammonia which was produced by its decomposition. Urease immobilized membrane was prepared by introducing anion-exchange group DEA into porous hollow-fiber membrane by radiation graft polymerization method, and immobilization of urease. When urease was immobilized at membrane introduced with anion-exchange group, the more increasing grafting rate, the more increasing immobilization amount. The result originates from the fact that a greater amount of protein was immobilized by forming a multilayer on the longer grafted chain. Meanwhile, the addition of the cross-linker was possible not only to suppress separation phenomenon produced during a washing process of immobilized urease membrane but also to enable the recycling of membrane. Urease Immobilized membrane with no separation phenomenon was prepared by cross-linking reaction for 5 h, and the hydrolysis rate of prepared urease immobilized membrane was over 98% and 50%, respectively, in 1 mol and 4 mol urea solutions.
Journal of the Korean Society of Marine Environment & Safety
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제25권2호
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pp.201-211
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2019
Safety and security measures in the shipping industry play a pivotal role in ensuring efficient and reliable cargo and passengers operations at each stage of the supply chain. The ISPS Code was adopted into SOLAS convention to protect seafarers and vessels from security threats. Furthermore, according to the Manila amendments to STCW Convention in 2010, personnel employed on board are required to participate in security training. Effective seafarers' education and training programs are of major importance to guarantee satisfactory performance levels onboard to minimize security-related risks. The study's contribution focuses on empirically evaluating the relationship between personal level of awareness and security performance when seafarers undertake security training courses. Findings of this study suggest that (1) seafarers who undertake maritime security training have a higher awareness of ship security, (2) security training and security awareness have a positive influence on security performance, and (3) security awareness mediates the impact of security training and security performance. In conclusion, education and training programs are key tools in enhancing seafarers' security awareness and security performance which, from an industry viewpoint, can translate into major economic, operational and reputational benefits.
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