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Detection of Campylobacter jejuni in food and poultry visors using immunomagnetic separation and microtitre hybridization

  • Simard, Ronald-E.
    • Proceedings of the Korean Society of Fisheries Technology Conference
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    • 2000.05a
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    • pp.71-73
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    • 2000
  • Campylobacter jejuni is most frequently identified cause of cause of acute diarrhoeal infections in developeed countries, exceeding rates of illness caused by both salmonella and shigilla(Skirrow, 1990 ; Lior 1994). Previous studies on campylobacter jejuni contamination of commercial broiler carcasses in u.s.(Stern, 1992). Most cases of the disease result from indirect transmission of Campylobactor from animals via milk, water and meat. In addition to Campylobactor jejuni. the closely relates species Campylobactor coli and Campylobactor lari have also been implicated as agents of gastroenteritis in humans. Campylobactor coli represented only approximately 3% of the Campylobactor isolates from patients with Campylobactor enteritis(Griffiths and Park, 1990) whereas Campylobactor coli is mainly isolated from pork(Lmmerding et al., 1988). Campylobactor jejuni has also been isolated from cases of bacteremia, appendicitis and, recently, has been associated with Guillai-Barre syndrome(Allos and Blaser, 1994; von Wulffen et al., 1994; Phillips, 1995). Studies in volunteers indicated that the infectious dose for Campylobactor jejuni is low(about 500 organisms)(Robinson, 1981). The methods traditionally used to detect Campylobactor ssp. in food require at least two days of incubation in an enrichment broth followed by plating and two days of incubation on complex culture media containing many antibiotics(Goossens and Butzler, 1992). Finnaly, several biochemical tests must be done to confirm the indentification at the species level. Therfore, sensitive and specific methods for the detection of small numbers of Campylobactor cells in food are needed. Polymerase chain reaction(PCR) assays targeting specific DNA sequences have been developed for the detection of Campylobactor(Giesendorf and Quint, 1995; Hemandex et al., 1995; Winter and Slavidk, 1995). In most cases, a short enrichment step is needed to enhance the sensitivity of the assay prior to detection by PCR as the number of bacteria in the food products is low in comparison with those found in dinical samples, and because the complex composition of food matrices can hinder the PCR and lower its sensitivity. However, these PCR systems are technically demanding to carry out and cumbersome when processing a large number of samples simutaneously. In this paper, an immunomagnetic method to concentrate Campylobactor cells present in food or clinical samples after an enrichment step is described. To detect specifically the thermophilic Campylobactor. a monoclonal antibody was adsorbed on the surface of the magnetic beads which react against a major porin of 45kDa present on the surface of the cells(Huyer et al., 1986). After this partial purification and concentration step, detection of bound cells was achieved using a simple, inexpensive microtitre plate-based hybridization system. We examined two alternative detection systems, one specific for thermophilic Campylobactor based on the detection of 23S rRNA using an immobilized DNA probe. The second system is less specific but more sensitive because of the high copy number of the rRNA present in bacterial cell($10^3-10^4$). By using specific immunomagnetic beads against thermophilic Campylobactor, it was possible to concentrate these cells from a heterogeneous media and obtain highly specific hybridization reactions with good sensitivity. There are several advantages in using microtitre plates instead of filter membranes or other matrices for hybridization techniques. Microtitre plates are much easier to handle than filter membranes during the adsorption, washing, hybridization and detection steps, and their use faciilitates the simultanuous analysis of multiple sample. Here we report on the use of a very simple detection procedure based on a monoclonal anti-RNA-DNA hybrid antibody(Fliss et al., 1999) for detection of the RNA-DNA hybrids formed in the wells.

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Antimicrobial Activity of Sodium chlorate and Korean Herbal Extracts against Mice infected with Escherichia coli O157:H7 (Escherichia coli O157:H7에 감염된 마우스에 대한 염소산나트륨과 한약재 복합추출물 합제의 항균효과)

  • Cha, Chun-Nam;Lee, Yeo-Eun;Son, Song-Ee;Park, Eun-Kee;Choi, Hyun-Ju;Kim, Suk;Lee, Hu-Jang
    • Journal of Food Hygiene and Safety
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    • v.27 no.1
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    • pp.81-86
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    • 2012
  • The present study was evaluated the antibacterial effect of the combination of $Coptidis$ $rhizoma$, $Glycyrrhiza$ $uralensis$ Fischet, $Schizandra$ $chinensis$ and $Corni$ $Fructus$(1:1:1) extracts(CGSC10). Furthermore, the effectiveness of CGSC10, sodium chlorate, and the combination of CGSC10 and sodium chlorate(CGSCS10) against $E.$ $coli$ O157:H7 infection was studied using ICR female mice. During the incubation period, the dose of 5, 10, and 20% CGSC10 was inhibited the growth of $E.$ $coli$ O157:H7 by 34.7, 60.2, and 76.4%, respectively. For 7 days after single challenge with $E.$ $coli$ O157:H7, forty female ICR mice were divided into four experimental groups which were administered in drinking water with saline, 10% CGSC10, 15 mM sodium chlorate, and CGSCS10, respectively. On the 3rd day, the number of $E.$ $coli$ O157:H7 in mouse feces was significantly decreased by administration of CGSC10, 15 mM sodium chlorate, and CGSCS10 ($p$ < 0.001). On the 7th day-after administration, CGSC10, sodium chlorate, and CGSCS10 were decreased the number of $E.$ $coli$ O157:H7 by 27.1, 67.7, and 83.3%, respectively. According to the results of the present study, administration of CGSCS10 to mice can reduce the severity of $E.$ $coli$ O157:H7 infection. In addition, it is suggested that CGSCS10 represents a good candidate for the treatment of enteric infections in domestic animals.

The Egg Development of Korean Slender Gudgeon, Squalidus gracilis majimae (Cypriniforms: Cyprinidae) (한국산 긴몰개 (Squalidus gracilis majime, Cyprinidae)의 난발생)

  • Park, Kyung-Seo;Hong, Young-Pyo;Moon, Woon-Ki;Choi, Shin-Suk;An, Kwang-Guk
    • Korean Journal of Ecology and Environment
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    • v.38 no.1 s.110
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    • pp.73-82
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    • 2005
  • This study was conducted, based on the field survey and laboratory observations, to elucidate egg developmental processes and their characteristics of the Korean slender gudgeon, Squalidus gracilis majimae. For the experiments, the mature adults were collected at the Woongcheon-Cheon Stream and Boreung Reservoir located in Boreung City, Chungnam Province and eggs were obtained from the natural spawning area. Morphological characteristics of the egg and embryonic development were summarized as follows: The shape of the fertilized egg was spherical, adhesive and transparent. The fertilized egg was 2.9${\pm}$0.3 mm (n = 30) in mean diameter under water temperature of $26{\pm}1.5^{\circ}C$, light white in color and had no oil droplets. After 20 minutes from the time of fertilization, a blastodisc was formed and divided into two cells at 48 minutes after fertilization. The blastular stage occurred at 5 hours 40 minutes after fertilization and the gastrular stage was detected at 8 hours 41 minutes after fertilization. The beginning of embryo formation was observed at 12 hours 58 minutes after fertilization and optic vesicles and 9 somites were discovered at 17 hours 05 minutes after fertilization. Differentiation of brains and embryo wiggling were observed at 37 hours 27 minutes after fertilization. Heart beating and the formation of melanophores in optic vesicles were detected at 44 hours 46 minutes after fertilization. The formation of pectoral fins and melanophores in the body were discovered at 50 hours 36 minutes after fertilization. Hatching occurred at 57 hours 49 minutes after fertilization. The newly hatched larvae were 3.3${\pm}$0.2 mm (n = 120) in total length. We believe that these results may contribute the species and population conservations under the situation of accelerated water pollution and the decreases of its diversity.

Clinical Characeristics of Intracordal Cysts (성대낭종의 임상적 특성)

  • Hong, Ki-Hwan;Park, Jung-Hoon;Kim, Won;Kim, Chang-Hyun
    • Journal of the Korean Society of Laryngology, Phoniatrics and Logopedics
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    • v.10 no.2
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    • pp.164-169
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    • 1999
  • Background and Objectives : The intracordal cysts are more increasingly diagnosed and treated due to advanced laryngeal stroboscopy and laryngeal microsurgical technique. The intracordal cysts are frequently misdiagnosed as vocal polyp or nodule The purpose of this study is to evaluate clinical features of intracordal cysts. Materials and Methods : In the present series, 83 cases of the intracordal cysts treated with laryngeal microsurgery are reported. The intracordal cysts are diagnosed preoperatively with indirect laryngoscopy, laryngeal endoscopy, laryngeal stroboscopy and confirmed with laryngeal microsurgical findings and biopsies. Results : Intracordal cysts are 83 of 1900 patients treated with laryngeal microsurgery(4.4%)-ductal cysts are 56 cases and epidermoid cysts are 27 cases. Intracordal cysts are more frequent in women, forties and the frequent site is an anterior third of the true vocal cord. With the indirect laryngoscopic examination, the ductal cysts are frequently misdiagnosed as vocal polyps or nodules but the epidermoid cysts are relatively easily diagnosed. The etiologic factors of the intracordal cysts are suspected as voice abuse and upper respiratory infection. The degree of postoperative voice satisfaction is similar to that of the vocal polyps. Conclusion : Intracordal cysts are frequently misdiagnosed as polyps or nodules, therefore preoperative stroboscopic findings and laryngeal microsurgical findings is important. An ideal treatment is to enucleate the cysts avoiding rupture of cyst and injury of lamina propria of the vocal cord.

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Comparison of High Resolution Image by Ortho Rectification Accuracy and Correlation Each Band (고해상도 영상의 정사보정 정확도 검증 및 밴드별 상관성 비교연구)

  • Jin, Cheong-Gil;Park, So-Young;Kim, Hyung-Seok;Chun, Yong-Sik;Choi, Chul-Uong
    • Journal of Korean Society for Geospatial Information Science
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    • v.18 no.2
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    • pp.35-45
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    • 2010
  • The objective of this study is to verify the positional accuracy by performing the orthometric corrections on the high resolution satellite images and to analyze the band correlation between the high resolution images corrected with orthometric correction. The objectives also included an analysis on the correlation of NDVI. For the orthometric correction of images from KOMPSAT2 and IKONOS, systematic errors were removed in use of RPC data, and non-planar distortions were corrected with GPS surveying data. Also, by preempting the image points at the same positions within ortho images, a comparison was performed on positional accuracies between image points of each image and GPS surveying points. The comparison was also made on the positional accuracies of image points. between the images. For correlation of band and correlation of NDVI, the descriptive statistics of DN values were acquired for respective bands by adding the Quickbird images and Aerial Photographs undergone through orthometric correction at the time of purchase. As result, from a comparison on positional accuracies of Orthoimages from KOMPSAT2 and Ortho Images of IKONOS was made. From the comparison the distance between the image points within each image and GPS surveying points was identified as 3.41m for KOMPSAT2 and as 1.45m for IKONOS, presenting a difference of 1.96m. Whereas, RMSE between image points was identified as 1.88m. The level of correlation was measured by using Quickbird, KOMPSAT2, IKONOS and Aerial Photographs between inter-image bands and NDVI, showing that there were high levels of correlation between Quickbird and IKONOS identified from all bands as well as from NDVI, except a high level of correlation that was identified between the Aerial Photographs and KOMPSAT2 from Band 2. Low levels of correlation were also identified between Quickbird and Aerial Photographs from Band 1. and between KOMPSAT2 and IKONOS from Band 2 and Band 4, whereas, KOMPSAT2 showed low correlations with Aerial Photographs from Band 3. For NDVI, KOMPSAT2 showed low level of correlations with both of QuickBird and IKONOS.

Analysis of consumers' needs and satisfaction related to food culture in Jeonju Hanok Village: Application of the Push-Pull factor theory (전주 한옥마을 음식문화에 대한 소비자 요구도 및 만족도 분석 : Push-Pull factor theory를 적용하여)

  • Na, Hee Ra;Park, Eun Ju;Yang, Soo Jin;Cha, Youn-Soo;Lee, Min A
    • Journal of Nutrition and Health
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    • v.50 no.2
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    • pp.192-200
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    • 2017
  • Purpose: The purposes of this study were to analyze visitors' food needs and identify the Push-Pull factor in Jeonju Hanok Village. Methods: A questionnaire was developed based on previous research to survey Korean adults who visited Jeonju city. A total of 580 questionnaires were used for the analysis. Results: Most of the subjects who visited Jeonju Hanok Village had food purchase experiences in Jeonju Hanok Village (96.4%). 'Traditional Korean food (26.5%)' was the most purchased food, followed by 'foreign food (25.8%)' and 'Korean food combined with foreign food (16.8%)'. Satisfaction of food purchases (3.35 points) was higher than average. The primary reason for satisfaction was 'the food is delicious (23.0%)', and the reason for dissatisfaction was 'the food is expensive (48.1%)'. In the push and pull factor analysis for identifying visit motivation, 'local food seeking', 'experience seeking', 'relaxation seeking', and 'friendship seeking' were push factors while 'traditional culture', 'facility convenience', 'experience activity', and 'food experience' were pull factors. There was a significant correlation between the push and pull factors. Regression analysis showed that all push factors influenced satisfaction. However, among pull factors, only 'food experience' and 'traditional culture' influenced satisfaction. Conclusion: In conclusion, this study indicates that food-related factors are destination characteristics (Pull factor) influencing intrinsic visit motivation (Push factor) and satisfaction. It is necessary to develop various food tourism products to satisfy visitors' needs and continuous visits in Jeonju Hanok Village. This study suggests the importance of research on food factors and provides useful basic data to establish positioning strategies for food cultural development in Jeonju Hanok Village.

Analysis and risk assessment of polycyclic aromatic hydrocarbons (PAHs) in seafood from oil contaminated bay (유류 오염지역의 수산물 중 다환방향족탄화수소류 (PAHs) 분석 및 위해평가)

  • Jeong, Ji-Yoon;Choi, Chan-Woong;Ryeom, Tai-Kyung;Cho, Kyoung-Hee;Park, Se-Ryung;Shin, Ho-Sang;Lee, Kwang-Ho;Lee, Hyo-Min
    • Analytical Science and Technology
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    • v.23 no.2
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    • pp.187-195
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    • 2010
  • To assess health risk for the intake among residents after the Hebei Spirit oil spill, 16 Polycyclic Aromatic Hydrocarbons (PAHs) in seafood samples from oil contaminated bay were determined by Gas Chromatography-Mass Spectrometry (GC-MSD) and samples were personally collected and purchased by residents. Samples were hydrolyzed with KOH and extracted with methylene chloride. The extracted solution were cleaned up using silica/florisil column and 16 PAHs were eluted by methylene chloride : n-hexane (1:9) mixture and determined by GC-MSD in Selected Ion Monitoring (SIM) mode. The mean recoveries for 16 PAHs ranged from 79% to 85%. The 16 PAHs levels in 126 samples ranged from 0.17 to $6.04\;{\mu}g$/kg and the TEQBaP (Toxic EQuivalents) levels in 126 samples were calculated using benzo(a)pyrene toxic equivalency factor for individual 16 PAHs and ranged from 0 to $0.91\;{\mu}gTEQ$/kg. The average Benzo(a)pyrene dietary exposure of residents was $5.5{\times}10^{-8}\;mg/kg$ bw/day and the average PAHs chronic dietary exposure was $1.3{\times}10^{-5}\;mg$ TEQ/kg bw/day. The margin of exposure (MOE) and the excess cancer risk and were $1.8{\times}10^6$ and $9.8{\times}10^{-8}$, respectively. Therefore, the assessment result was considered as low concern for health risk.

Development and validation of an analytical method for nematicide imicyafos determination in agricultural products by HPLC-UVD (HPLC-UVD를 이용한 살선충제 imicyafos의 시험법 개발 및 검증)

  • Do, Jung-Ah;Park, Hyejin;Kwon, Ji-Eun;Choi, Won-Jo;Lee, Hyun-Sook;Chang, Moon-Ik;Hong, Jin-Hwan;Oh, Jae-Ho
    • Analytical Science and Technology
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    • v.27 no.5
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    • pp.234-242
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    • 2014
  • Imicyafos which is a nematicide for controlling root-knot nematodes has been registered in the Republic of Korea in 2012, and the maximum residue limits of imicyafos are set to watermelon and korean melon as each 0.05 mg/kg. Extremely reliable and sensitive analytical method is required for ensuring food safety on imicyafos residues in agricultural commodities. Imicyafos residues in samples were extracted with acetone, partitioned with hexane and dichloromethane, and then purified with florisil. The purified samples were analyzed by HPLC-UVD and confirmed with LC-MS. Linear range was between 0.1~5 mg/kg with the correlation coefficient ($r^2$) 0.99997. Average recoveries of imicyafos ranged from 77.0 to 115.4% at the spiked levels of 0.02 and 0.05 mg/kg with the relative standard deviations of 2.2~9.6%. Limit of detection and quantification were 0.005 and 0.02 mg/kg, respectively. An inter-laboratory study was conducted to validate the determination method in depth, and the results were satisfactory. All of the validation results revealed that the developed analytical method in this study is relevant for imicyafos determination in agricultural commodities and will be used as an official analytical method.

Erythrocyte Manganese Superoxide Dismutase Activity Indicates Training Intensity for Racing Horses (적혈구의 Manganese Superoxide Dismutase 활성은 경주마의 훈련강도를 나타낸다)

  • Choi, Jun-Young;Park, In-Kyung;Im, Jin-Taek;Koh, Tae-Song
    • Journal of Animal Science and Technology
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    • v.50 no.4
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    • pp.573-580
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    • 2008
  • This study was aimed to investigate that training of horses is related with the activity of superoxide dismutase(SOD) in erythrocyte of racing horses. The SOD activity was assayed from erythrocyte of six Thoroubred horses having final stage of training, about 21 month-old, 474~509 kg body weight for race trainig. During 7 weeks of training period from 24th Sep. to 6th Nov, horses were bled very carefully 4 times at 1st Oct, 16th Oct, 30th Oct. and 6th Nov. As the training period passed, erythrocyte of the horses have gradually increased the MnSOD activity(p<0.05) and lowered the CuZnSOD activity. The plasma ceruloplasmin and peroxidase activities, and lactate levels were reduced gradually while peroxide and glucose levels gradually increased. The calculated oxygen consumption(Eaton, 1995) for training of horses were linearly related with the MnSOD activity(r=0.650, n=32) but negatively with CuZnSOD activity in erythrocyte and lactate levels(r=-349, n=32) in plasma. Also, peroxide levels in plasma of horses had positive relation with the MnSOD activity in erythrocyte(r=0.616, n=48). In conclusions, as the training is progressed, the raised MnSOD activity in erythrocytes and peroxide levels in plasma indicated balances between oxidant and antioxidants for the protection from ROS during race of horses. The results showed that the MnSOD activity in erythrocyte and peroxide levels in plasma may be used as marker for the intensity of training racing horses.

Dietary Brown Seaweed Modulates Nutrient Metabolism, Anti-oxidant System and Immune Response in Broiler Chicks Injected LPS i.p. (사료 중 미역은 복강내 LPS 주입 육계 병아리에서 영양소 대사, 항산화계 및 면역 반응을 조정한다)

  • Park, In-Kyung;Im, Jin-Taek;Choi, Do-Yul;Koh, Tae-Song
    • Journal of Animal Science and Technology
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    • v.50 no.2
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    • pp.185-198
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    • 2008
  • Influences of dietary brown seaweed(BSW) on the nutrient metabolism, anti-oxidant enzyme activity and cell-mediated immune response were studied in broiler chicks activated acute phase response. 72 Hatched male broiler chicks(Ross) were divided into 12 pens, 6 heads per pen, and fed the BSW 0.0% (Basal) or 2.0% diet, respectively, and injected with the Salmonella typhimurium lipopoly saccharide(LPS) for activation of the acute phase response three times at 8, 10 and 12 d of age. During 4 wks of experimental feeding, growth performance of broiler chicks was not affected by dietary BSW and the acute phase response. Compared with control birds, the acute phase response did not affect the daily weight gain in birds fed BSW 2.0% diet, decreased nitrogen balance(NB) or metabolizable energy(ME) utilization per metabolic body size(kg0.75), and enhanced activities of peroxidase or extracellular SOD(EcSOD), tumor necrosis factor-alpha and ovotransferrin in plasma and MnSOD and CuZnSOD in erythrocyte cytosol. Compared to BSW 0.0% diet, 2.0% diet enhanced protein retention(NB) per kg0.75 regardless the acute phase response, did not affect uric acid nitrogen excretion(UAN) per kg0.75 in birds during the acute phase response, decreased(p<0.05) the UAN excretion per kg0.75 in control birds. And BSW 2.0% diet also decreased(p<0.05) plasma peroxide level and erythrocyte peroxidase or MnSOD activity but increased plasma peroxidase and EcSOD activity and interleukin-1 activity secreted from LPS-stimulated PBMC in 4 week broiler chicks.