• Title/Summary/Keyword: Cross contamination

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A Rapid and Sensitive Detection of Aflatoxin-producing Fungus Using an Optimized Polymerase Chain Reaction (PCR)

  • Bintvihok, Anong;Treebonmuang, Supitchaya;Srisakwattana, Kitiya;Nuanchun, Wisut;Patthanachai, Koranis;Usawang, Sungworn
    • Toxicological Research
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    • v.32 no.1
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    • pp.81-87
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    • 2016
  • Aflatoxin B1 (AFB1) is produced by Aspergillus flavus growing in feedstuffs. Early detection of maize contamination by aflatoxigenic fungi is advantageous since aflatoxins exert adverse health effects. In this study, we report the development of an optimized conventional PCR for AFB1 detection and a rapid, sensitive and simple screening Real-time PCR (qPCR) with SYBR Green and two pairs of primers targeting the aflR genes which involved aflatoxin biosynthesis. AFB1 contaminated maize samples were divided into three groups by the toxin concentration. Genomic DNA was extracted from those samples. The target genes for A. flavus were tested by conventional PCR and the PCR products were analyzed by electrophoresis. A conventional PCR was carried out as nested PCR to verify the gene amplicon sizes. PCR-RFLP patterns, obtained with Hinc II and Pvu II enzyme analysis showed the differences to distinguish aflatoxin-producing fungi. However, they are not quantitative and need a separation of the products on gel and their visualization under UV light. On the other hand, qPCR facilitates the monitoring of the reaction as it progresses. It does not require post-PCR handling, which reduces the risk of cross-contamination and handling errors. It results in a much faster throughout. We found that the optimal primer annealing temperature was $65^{\circ}C$. The optimized template and primer concentration were $1.5{\mu}L\;(50ng/{\mu}L)$ and $3{\mu}L\;(10{\mu}M/{\mu}L)$ respectively. SYBR Green qPCR of four genes demonstrated amplification curves and melting peaks for tub1, afIM, afIR, and afID genes are at $88.0^{\circ}C$, $87.5^{\circ}C$, $83.5^{\circ}C$, and $89.5^{\circ}C$ respectively. Consequently, it was found that the four primers had elevated annealing temperatures, nevertheless it is desirable since it enhances the DNA binding specificity of the dye. New qPCR protocol could be employed for the determination of aflatoxin content in feedstuff samples.

Microbiological Hazard Analysis of Ginseng Farms at the Cultivation Stage to Develop a Good Agricultural Practices (GAP) Model (인삼의 GAP 실천모델 개발을 위한 재배단계의 미생물학적 위해도 평가)

  • Shim, Won-Bo;Kim, Jeong-Sook;Chung, Duck-Hwa
    • Journal of Food Hygiene and Safety
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    • v.28 no.4
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    • pp.312-318
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    • 2013
  • This study validated microbiological hazards of ginseng farms at the cultivation stage and suggested recommendations to develop a good agricultural practices (GAP) model. A total of 96 samples were collected from cultivation environments (soil, irrigation water, and atmosphere), plants (ginseng and its leaf), personnel hygiene (glove, cloth, and hand) of 3 ginseng farms (A, B, and C) and were tested to analyze sanitary indicator bacteria (aerobic plate count, coliforms and Escherichia coli), major foodborne pathogens (E. coli O157:H7, Listeria monocytogenes, Salmonella spp., Staphylococcus aureus, and Bacillus cereus), and fungi. Total bacteria, coliform, and fungi in the 3 ginseng farms were detected at the level of 1.3~6.0, 0.1~5.0, and 0.4~4.9 v/g (or mL, hand, and $100cm^2$), respectively. Only irrigation water collected from one ginseng farm was confirmed to be E. coli positive. In case of pathogenic bacteria, B. cereus was detected at levels of 0.1~5.0 log CFU/g (or mL, hand, and $100cm^2$) in all samples, but other pathogen bacterias were not detected in any samples from all farms. Although E. coli were detected in irrigation water, the level of microbial for the three farms was lower than the regulation limit. According to the results, the ginsengs produced from the 3 farms were comparatively safe with respect to microbiological hazard. However, cross-contamination of bacteria from environments and workers to ginseng has been considered as potential risks. Therefore, to minimize microbial contamination in ginseng, GAP model should be applied for ensuring the safety of ginsengs.

Determination of fatty acid methyl esters (FAME) content in aviation turbine fuel using multi-dimensional GC-MS (Multi-dimensional GC-MS를 이용한 항공터빈유의 FAME 함량 분석)

  • Youn, Ju Min;Doh, Jin Woo;Hwang, In Ha;Kim, Seong Lyong;Kang, Yong
    • Journal of the Korean Applied Science and Technology
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    • v.34 no.4
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    • pp.717-726
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    • 2017
  • The current allowable cross-contamination level of fatty acid methyl esters (FAME) in aviation turbine fuel (AVTUR) is 50 mg/kg, due to that the presence of FAME in AVTUR can significantly impact the fuel supply system and jet engine. It has been difficult to analyze the level of FAME in AVTUR, since it is consisted of a lot of hydrocarbons. In this study, thus, a new method using multi-dimensional GC-MS (MDGC-MS) was proposed in order to determine the FAME level in AVTUR effectively. Applying to MDGC-MS with Deans switching system enabled us to detect and quantify the FAME with low carbon numbers such as those derived from coconut oil and palm kernel oil. The matrix effect of MDGC-MS method, which could shift the FAME peaks to slightly longer retention times, was reduced by 20 times compared with that of 1-dimensional GC-MS reference method. This developed method could be suitable for qualitative and quantitative analyses to determine the contamination level of trace FAME in AVTUR.

Toxin Genes and Antibiotic Resistance of Food Poisoning Bacteria Isolated from Food Service Equipment in Childcare Centers (어린이집 급식설비에서 분리된 식중독 미생물의 독소 유전자 및 항생제 내성)

  • Eun-yeong Kim;Chae-Young Kim;Ji-Yu Im;Jung-Beom Kim
    • Journal of Food Hygiene and Safety
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    • v.39 no.3
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    • pp.266-272
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    • 2024
  • This study assessed the contamination level of food poisoning bacteria on handles of food service equipment in childcare centers to prevent food poisoning and analyzed toxin genes and antibiotic resistance of isolated strains. The isolates used in this study were collected from 101 childcare centers in Jeollanam-do. Four strains of Bacillus cereus and two strains of Staphylococcus aureus were isolated on the handles of food service equipment (refrigerators and freezers). The toxin genes of B. cereus were detected as nheA, nheB, nheC, entFM, and cytK. No toxin genes of S. aureus were detected. B. cereus showed resistance to β-lactam antibiotics, such as ampicillin and cefepime. S. aureus also showed resistance to antibiotics such as ampicillin and cefepime. Therefore, microbial safety and hygiene management, such as periodic sterilization of handles, should be strengthened to prevent food poisoning caused by cross-contamination of food service equipment handles in childcare centers.

The Influence of Various Factors upon the Membrane Filter Technique on Raw Water of the Nak-Dong River (낙동강 원수에 대한 대장균군 막여과시험법에 있어서 여러 인자가 결과에 미치는 영향)

  • Hyun, Jae-Yeoul;Yoon, Jong-Ho;Shin, Sang-Hee;Kim, Jong-Woo
    • Journal of Korean Society on Water Environment
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    • v.25 no.2
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    • pp.205-211
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    • 2009
  • In this study, the membrane filter method was compared to the MPN method for the analysis of total coliforms from raw water using raw test waters and controls including 6 standard strains of coliforms, and the various factors were analyzed on the detection of general and fecal origin coliforms. The range of error rate for the detection of 5 standard strains using the membrane filter and the MPN methods was 0 to 6% and 45 to 133%, respectively. The error rate of the membrane filter method was lower than that of the MPN method. The membrane filter method (m-Endo) showed 10% (11 out of 111) of difference for the detection sensitivity of coliforms isolated from raw water compared to the MPN method (BGLB). The membrane filter method was less affected by the factors including temperature, turbidity, charcoals of powder form, contamination, and reverse pressure. In conclusion, the membrane filter method is a better method for the analysis of total coliforms from raw water than the MPN method, considering the accuracy of detection and the tolerance to various experimental factors.

우리나라에서 보고된 집단 식중독의 발생 특징(1981-1989)

  • 홍종해;이용욱
    • Food Industry
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    • s.109
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    • pp.34-40
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    • 1991
  • 1. 환자발생 규모는 10명 이하가 $38.0\%$로 가장 많았고 50명 이상의 대규모 발생도 $13.0\%$이었다. 대규모 환자발생은 세균성 식중독이 원인이었다. 2. 전체 식중독에 $67.3\%$가 7월부터 9월에 집중되어 계절의 변화에 영향받고 있음을 알 수 있다. Vibrio에 의한 발생은 특히 하절기에 높았고 Salmonella, Staphyloccus는 상대적으로 연중 계속해서 발생 가능성을 보였다. 독성물질에 의한 식중독은 계절과 무관하였으나 식물성 식중독은 7, 8월에 집중되고 있다. 3. 세균성 식중독이 $56.2\%$, 독성물질에 의한 식중독이 $17.8\%$이었다. 세균성 식중독의 원인균은 Vibrio $35.4\%$, Salmonella $27.2\%$, Escheri-chia coli $17.7\%$, Staphylococcus $17.7\%$ 순이었다. 독성물질에 의한 식중독은 식물성 식중독이 $64\%$로 가장 많았다. 4. Vibrio 식중독의 주원인식품은 해산물이지만 cross-contamination에 의해 오염된 다른 식품으로도 발생한 것으로 나타났다. Salmone-lla 식중독은 관혼상제시 거의 빠지지 않고 제공되는 돼지고기 편육이 주원인식품이었다. 5. 전체 식중독의 $76.8\%$가 가정과 식품접객업소(구내식당 포함)에서 발생되었으며, 특히 도시지역은 $66.4\%$가 식품접객업소에서 농촌지역은 $58.9\%$가 가정에서 발생되어 지역간의 차이를 보였다. 6. 독버섯은 독성물질에 의한 식중독의 주원인식품으로 농촌가정에서 일상식을 통한 섭취로 많이 발생되고 있으며, 사망자의 발생이 가장 많은 치명적인 원인식품이었다. 7. 식품접객업소에서 발생된 식중독의 주원인식품은 해산물이며, 덜익혀지거나 생선회 같이 날것을 섭취하여 발생하였다. 8. 생일잔치 및 회갑연, 결혼 피로연, 문상, 제사 등의 관혼상제시 발생되는 식중독은 농촌지역의 가정에서 주로 발생되고 있다. 구내 식당 및 관광지 음식점의 식중독 발생이 많은 것은 이들 시설에 대한 위생관리가 미비하여 대규모 식중독 발생의 위험이 항시 존재하고 있다고 할 수 있다.

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A Study on the Hygiene Practices of Foodservice Employees by Hygiene Education and Work Environment in the Gyeongnam Area (경남지역 위탁급식업체 조리종사자의 위생교육 및 근무환경에 따른 위생 직무수행도 평가)

  • Lim, Jung-Hee;Kim, Hyun-Ah;Jung, Hyun-Young
    • Journal of the Korean Dietetic Association
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    • v.19 no.3
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    • pp.209-222
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    • 2013
  • This study examined the hygiene practices of contract foodservice employees and investigated the influence of education and work environment on these hygienes practices. A questionnaire was distributed to 250 contract foodservice employees and a total of 232 responses were received and analyzed. The overall score for hygiene practices of contract foodservice employees was 3.89 based on a 5-point scale. Cross-contamination was prominent (highest score at 4.46) and the heating temperature was less prominent (lowest score at 3.49). The factors most affecting contract foodservice employees were their work period, the size of the contract foodservice management company, the number of meals served daily, the frequency of meal service per day and the frequency of hygiene education. Compared to small and medium-sized contract foodservice management companies, the major contract foodservice management companies showed higher scores for refrigerator/freezer control (P<0.001), vegetable/fruit disinfection (P<0.001), thawing (P<0.001), heating temperature (P<0.001), cleaning/disinfection (P<0.01), and personal hygiene (P<0.05). The frequency of hygiene education had a significant effect on the performance levels for refrigerator/freezer control (P<0.001), vegetable/fruit disinfection (P<0.001), thawing (P<0.001), heating temperature (P<0.001), cleaning/disinfection control (P<0.001), food supply control (P<0.05), and personal hygiene (P<0.05). From these results, to increase the sanitation quality of contract foodservice operations, hygiene practice levels need to increase and hygiene education systematically should be enforced for foodservice employees.

Practical application of DNA markers for high-throughput authentication of Panax ginseng and Panax quinquefolius from commercial ginseng products

  • Jung, Juyeon;Kim, Kyung Hee;Yang, Kiwoung;Bang, Kyong-Hwan;Yang, Tae-Jin
    • Journal of Ginseng Research
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    • v.38 no.2
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    • pp.123-129
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    • 2014
  • Korean ginseng (Panax ginseng) and American ginseng (Panax quinquefolius) are widely used medicinal plants with similar morphology but different medicinal efficacy. Roots, flowers, and processed products of Korean and American ginseng can be difficult to differentiate from each other, leading to illegal trade in which one species is sold as the other. This study was carried out to develop convenient and reliable chloroplast genome-derived DNA markers for authentication of Korean and American ginseng in commercial processed products. One codominant marker could reproducibly identify both species and intentional mixtures of the two species. We further developed a set of species-unique dominant DNA markers. Each species-specific dominant marker could detect 1% cross contamination with other species by low resolution agarose gel electrophoresis or quantitative polymerase chain reaction. Both markers were successfully applied to evaluate the original species from various processed ginseng products purchased from markets in Korea and China. We believe that high-throughput application of this marker system will eradicate illegal trade and promote confident marketing for both species to increase the value of Korean as well as American ginseng in Korea and worldwide.

Study on the Characteristics of Food-borne Pathogens Isolated from Students' Mobile Phones in Busan (부산지역 중·고등학생 휴대전화에서 분리한 식중독균의 특성 연구 - 대장균, 황색포도상구균, 바실러스 세레우스를 중심으로 -)

  • Park, Sun-Hee;Park, Yeon-Kyoung;Hwang, In-Yeong;Park, Hye-Young;Sung, Gyung-Hye;Jo, Hyeon-Cheol
    • Journal of Environmental Health Sciences
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    • v.42 no.4
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    • pp.255-265
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    • 2016
  • Objectives: Mobile phones have become one of the most essential accessories in daily life. However, they may act as reservoir of infectious pathogens if they are used without hygienic practices in their handling. Therefore, this study aimed to isolate food-borne pathogens from mobile phones and investigate the characteristics of toxin genes and antibiotic susceptibility patterns. Methods: A total of 146 mobile phones were collected from 83 middle- and 63 high-school students in Busan. The surfaces of the mobile phones were aseptically swabbed. Results: Among the food-borne pathogens, Staphylococcus aureus, Bacillus cereus and Escherichia coli were detected in 26 (17.8%), 20 (13.7%) and four (2.7%) samples, respectively. There were no statistically significant differences according to school level, gender or phone type. None of four E. coli strains had pathogenic toxic genes. All of the B. cereus strains carried at least three different toxin genes among the nine enterotoxin and emetic toxin genes. Three out of 20 B. cereus strains (15%) possessed emetic toxin genes, which are rarely detected in food-poisoning cases in Korea. Among the 26 strains of S. aureus, the detection rate of staphylococcal enterotoxin genes, toxic shock syndrome toxin (tsst) and factors essential for methicillin resistance (femA) were 84.6%, 7.7% and 100%, respectively. In the antibiotic susceptibility test, there was no methicillin-resistant S. aureus (MRSA) or vancomycin-resistant S. aureus (VRSA). Conclusion: The results show that students' mobile phones in Busan were contaminated by food-borne pathogens which carried various toxic genes. Therefore, regular phone disinfection and hand hygiene is important in order to reduce cross-contamination.

EFFECT ON IMMERSI0N DISINFECTION OF HYDROPHIILIC RUBBER IMPRESSI0N MATERIAL ON DIMENSIONAL STABILITY AND SURFACE HARDNESS OF IMPROVED STONE CAST (친수성 고무인상재의 침적 소독이 경석모형의 크기의 안정성과 표면경도에 미치는 영향)

  • Nam, Mee-Hyun;Kang, Woo-Jin;Chung, Moon-Kyu
    • The Journal of Korean Academy of Prosthodontics
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    • v.33 no.3
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    • pp.569-583
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    • 1995
  • Disinfection of dental impressions are necessary due to contamination with patient’s saliva and blood, which is a potential for cross-infection. The purpose of this study was to evaluate the effects of disinfection of four hydrophilic rubber impression materials with three disinfecting solutions, on the dimensional stability and surface hardness of improved stone casts. Three hydrophilic vinyl polysiloxane impression materials(Express, Reprosil, Exafine) and one polyether impression material(Impregum-F) were mixed according to the manufacturer’s directions and impressions were made on a ADA specification No. 19 stainless-steeldie. On removal of the impressions, each impression was immersed in one of the disinfectants(Banicide, Potadine, Clorox) for 10 minutes. After disinfection, type IV improved stone. casts were poured. On this cast, the linear dimension and surface hardness were measuredusing a Measurescope(Nikon, Japan) and a Barcol hardness tester(Barber, Colman Co U. S. A). The results were as follows : 1. The improved stone casts from disinfected Reprosil and Impregum-F impression material did not show dimensional changes(P>0.01). Those from disinfected Express and Exafine impression material showed dimensional changes(P<0.01). The amount of shrinkage was not clinically significant. 2. The improved stone casts from disinfected Express impression material did not exhibit changes in surface hardness(P>0.01), but those from disinfected Reprosil, Exafine, Imp regnum-F impression material showed changes in surface hardness(P<0.01). 3. The dimensinal stability and surface hardness of the improved stone casts were satisfactory using Banicide on Express, all disinfectants used in this study on Reprosil, Potadin and Clorox on Exafine, Banicide and Clorox on Impregum-F. According to these results, immersion disinfection of hydrophilic rubber impression mate rials did not adversely affect the resultant casts. Nevertheless compatibility tests of impression materials and disinfectants should be done when disinfecting impressions.

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