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Determination and Survey of Fluoroquinolones Residue in Chicken Muscle by HPLC with Fluorescence Detector (액체크로마토그래피-형광검출기를 이용한 닭고기 중 플루오로퀴놀론계 항균물질 정량분석 및 잔류조사)

  • 박은정;임지흔;이성모
    • Journal of Food Hygiene and Safety
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    • v.19 no.1
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    • pp.12-18
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    • 2004
  • Ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin in chicken muscle were seperated by liquid extraction and determined with high performance liquid chromatography (HPLC) with fluorescence detector. Analysis was carried out using following conditions; Cl8 column (250${\times}$4.6 mm i.d. 5 ${\mu}{\textrm}{m}$ particle size), mobile phase composed of D.W. (containing 0.4% triethylamine and phospholic acid): methanol : acetonitrile (800:100:100, v/v/v), isocratic pump at a flow rate of 1.0 $m\ell$/min and 50 ${mu}ell$ of injection volume, fluorescence detector with EX278 nm/EM.456 nm. The calibration curves of four fluoroquinolones showed linearity (${\gamma}$$^2$$\geq$0.999) at concenration range of 0.025-0.6 $\mu\textrm{g}$/ml. The recoveries in fortified chicken muscle represented more than 80% with low coefficient of variation (〈10%) for concentration range of four fluoroquinolones. The detection limits for ofloxacin, norfloxacin, ciprofloxacin, and enrofloxacin were 23.5, 3.4, 3.0 and 2.5 ng/g in chicken muscle, respectively. We also monitored fluoroquinolones residue in muscle of chickens (broiler 1:227, Korean native chicken 219, laying chicken 77) using EEC-4-plate screening and HPLC conformation methods. Ten(broiler 5, Korean native chicken 5) out of the fifteen samples which were positively detected by EEC-plate screening method from 1,523 chicken meat were confirmed with ciprofloxacin and enrofloxacin by HPLC. The ranges of residual concentration were 0-0.12 ppm for ciprofloxacin and 0.01-6.79 ppm for enrofloxacin. In conclusion, our method could be applied effectively to determine four fluoroquinolones residues in chicken meat, and further survey for fluoroquinolones residue in chicken meat are needed for more effective control of fluoroquinolones used in livestock.

Effects of Small Scale Post-Harvest Facility and Hygiene Education on the Level of Microbial Safety in Korean Leeks Production (영양부추 생산농가의 소규모 수확후 처리시설 적용과 위생교육에 따른 미생물학적 안전성 향상 효과)

  • Kim, Se-Ri;Kim, Jin-Bae;Lee, Hyo-Sup;Lee, Eun-Sun;Kim, Won-Il;Ryu, Song-Hee;Ha, Jihyung;Kim, Hwang-Yong;Ryu, Jae-Gee
    • Journal of Food Hygiene and Safety
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    • v.30 no.3
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    • pp.249-257
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    • 2015
  • The purposes of this study were to develop a small scale post-harvest facility, and consequently to evaluate the effects of applying the facility along with hygiene education on the level of microbial safety in Korean leeks production. A total of 135 samples were collected at three Korean leeks farms in Yangju, Gyeonggi province. Food safety indicators (Aerobic plate count (APC), coliform count, and Escherichia coli) and foodborne pathogens (E. coli O157:H7, Salmonella spp., Staphylococcus aureus, Listeria monocytogenes, and Bacillus cereus) on/in the samples were assessed. The microbial load measured as APC with harvesting tools such as comb, chopping board, and knife, at the farms where the small scale post-harvest facility had been operated (Farms A and B) was lower than that at another farm having no post-harvest facility (Farm C) by 1.44~2.33 log CFU / $100cm^2$. Moreover, the chopping board from Farm C was observed being contaminated with B. cereus at 6.03 log CFU / $100cm^2$. The coliform counts from the samples increased by 0.57~1.89 log CFU/g after leeks was submerged in ground water for washing. E. coli was recovered from leeks, soil, and the ground water used in the washing process, while no E. coli O157:H7, Salmonella spp., and L. monocytogenes was detected. Our results indicated that the small scale post-harvest facility developed in this study as well as the hygiene education played an important role in enhancing the level of microbial food safety in the leeks production environment. However, a disinfection technique could be needed during the washing step in order to prevent a potential contamination.

Microbial Hazard Analysis of Manufacturing Processes for Starch Noodle (당면의 제조공정별 미생물학적 위해요소 분석)

  • Cheon, Jin-Young;Yang, Ji Hye;Kim, Min Jeong;Lee, Su-Mi;Cha, Myeonghwa;Park, Ki-Hwan;Ryu, Kyung
    • Journal of Food Hygiene and Safety
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    • v.27 no.4
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    • pp.420-426
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    • 2012
  • The purpose of this study was to identify control points through microbiological hazard analysis in the manufacturing processes of starch noodles. Samples were collected from the ingredients, manufacturing processes, equipment and environment. Microbiological hazard assessments were performed using aerobic plate counts (APC), Enterobacteriaceae (EB), E. coli and five pathogens including B. cereus, E. coli O157:H7, L. monocytogenes, Salmonella spp., and S. aureus. The APC levels in raw materials were from 2.12 to 3.83 log CFU/g. The contamination levels after kneading were 4.31 log CFU/g for APCs and 2.88 log CFU/g for EB counts. APCs decreased to 1.63 log CFU/g and EB were not detected after gelatinization, but their levels slightly increased upon cooling, cutting, ripening, freezing, thawing, and separating. The reuse of cooling and coating water would be a critical source of microbial increase after cooling. After drying, APCs and EB counts decreased to 5.05 log CFU/g and 2.74 log CFU/g, respectively, and the levels were maintained to final products. These results suggest that the cooling process is a critical control point for microbiological safety, and the cooling water should be treated and controlled to prevent cross contamination by pre-requisite program.

Validation of PCR and ELISA Test Kits for Identification of Domestic Animal Species in Raw Meat and Meat Products in Korea (국내 유통 식육 및 식육가공품에서 축종감별을 위한 PCR 및 ELISA 검사법 검증)

  • Heo, Eun-Jeong;Ko, Eun-Kyung;Seo, Kun-Ho;Kim, Young-Jo;Park, Hyun-Jung;Wee, Sung-Hwan;Moon, Jin-San
    • Journal of Food Hygiene and Safety
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    • v.29 no.2
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    • pp.158-163
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    • 2014
  • In this study, two commercial PCR and ELISA test kits were examined for identification of eight animal species (beef, pork, chicken, duck, turkey, goat, lamb, and horse) from raw meat and meat products in Korea. The detection limit in RAW meat ELISA kit$^{(R)}$ on three types of meat samples blended with beef, pork and chicken, demonstrated that all meat species were differentiable down to 0.2%. RAW meat ELISA kit$^{(R)}$ on animal species resulted in differentiation rate of 94.5% for beef, 93.3% for pork, 90% for lamb, and 100% for chicken, duck, turkey, goat, and horse. In contrast, Powercheck Animal Species ID PCR kit$^{TM}$ resulted in 100% specificity at 0.05% limit of detection for all meat species. The detection limit of Cooked Meat ELISA kit$^{(R)}$ on mixed meat samples heat-treated with different temperatures and times, resulted in 0.1% for all heat-treated mixed meat except for chicken at 1.0%. Additionally, ELISA kit on sixty meat products resulted in specificity of 31.8% for ham, 13.6% for sausages, and 12.5% for ground processed products, and relatively low rate for more than 2 types of mixed meats. On the contrary, meat species differentiation using PCR kit showed higher percentage than that using ELISA kit$^{(R)}$: 50.0% for ham, 41.7% for sausages, and 28.6% for ground processed meat. Futhermore, PCR kit on 54 dried beef meats detected pork genes in 13 products whereas ELISA kit showed negative results for all products. Hence, the possibility of cross-contamination during manufacturing process was investigated, and it was found that identical tumblers, straining trays, cutters and dryers were used in both beef and pork jerky production line, suggesting the inclusion of pork genes in beef products due to cross-contamination. In this study, PCR and ELISA test kits were found to be excellent methods for meat species differentiation in raw meat and heat-processed mixed meat. However, lower differentiation rate demonstrated in case of meat processed products raised the possibility of inclusion of other species due to cross-contamination during manufacturing process.

Development of Species-Specific PCR to Determine the Animal Raw Material (종 특이 프라이머를 이용한 동물성 식품원료의 진위 판별법 개발)

  • Kim, Kyu-Heon;Lee, Ho-Yeon;Kim, Yong-Sang;Kim, Mi-Ra;Jung, Yoo Kyung;Lee, Jae-Hwang;Chang, Hye-Sook;Park, Yong-Chjun;Kim, Sang Yub;Choi, Jang Duck;Jang, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.347-355
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    • 2014
  • In this study, the detection method was developed using molecular biological technique to distinguish authenticity of animal raw materials. The genes for distinction of species about animals targeted at Cytochrome c oxidase subunit I (COI), Cytochrome b (Cytb), and 16S ribosomal RNA (16S rRNA) genes in mitochondrial DNA. The species-specific primers were designed by that Polymerase Chain Reaction (PCR) product size was around 200 bp for applying to processed products. The target 24 raw materials were 2 species of domestic animals, 6 species of poultry, 2 species of freshwater fishes, 13 species of marine fishes and 1 species of crustaceans. The results of PCR for Rabbit, Fox, Pheasant, Domestic Pigeon, Rufous Turtle Dove, Quail, Tree Sparrow, Barn Swallow, Catfish, Mandarin Fish, Flying Fish, Mallotus villosus, Pacific Herring, Sand Lance, Japanese Anchovy, Small Yellow Croaker, Halibut, Jacopever, Skate Ray, Ray, File Fish, Sea Bass, Sea Urchin, and Lobster raw materials were confirmed 113 bp ~ 218 bp, respectively. Also, non-specific PCR products were not detected in compare species by species-specific primers. The method using primers developed in this study may be applied to distinguish an authenticity of food materials included animal raw materials for various processed products.

Detection and Differentiation of Intentional and Unintentional Mixture in Raw Meats Using Real-time PCR (Real-time PCR을 이용한 식육원료의 의도적, 비의도적 혼입 판별법 개발)

  • Kim, Kyu-Heon;Kim, Mi-Ra;Park, Young-Eun;Kim, Yong-Sang;Lee, Ho-Yeon;Park, Yong-Chjun;Kim, Sang Yub;Choi, Jang Duck;Jang, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.340-346
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    • 2014
  • In this study, the detection method was developed using real-time PCR to distinguish 4 species (bovine, porcine, horse, and chicken) of raw meats. The genes for distinction of species about meats targeted at 12S rRNA and 16S rRNA parts in mitochondrial DNA. Probes were designed to have a 5' FAM and a TAMRA at the 3' end. This study is to develop 4 species-specific primer and probes about raw materials and real-time PCR on 10 strains to observe the products of non-specific signal for similar species. As a result, any non-specific signal were not detected among each other. Real-time PCR method was developed for quantitation and identification of intentional and unintentional mixture in ground mixed meat (The difference of $C_T$ value between intentional mixture and 100% meat: $${\leq_-}$$ cycles, The difference of $C_T$ value between unintentional mixture and 100% meat: $${\geq_-}$$ cycles). The detection and differentiation of intentional and unintentional mixture in this study would be applied to food safety management for eradication of adulterated food distribution and protection of consumer's right.

Monitoring of Microorganism Contamination in Children-Preferred Confectioneries in Korea (서울·경인지역 유통판매중인 과자류의 위생지표 세균 및 식중독균 오염도 조사)

  • Koo, Eun Joo;Chung, So Young;Park, Ji Eun;Kwon, Yu Jihn;Seo, Dong Hyuk;Jung, Yu Young;Cho, Kyong Chul;Lee, Yo A;Min, Hee Eun;Kim, Eu Gene;Kim, Hyun Jung;Kim, Seul Ki;Choi, Sun Ok;Lim, Chul Ju
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.322-326
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    • 2014
  • According to Centers for Disease Control and Prevention (CDC, USA) recently it was reported that the children (< 5 year-old children) were more susceptible to Foodborne-illness. Confectionery products should be strictly controlled because they are children-preferred foods. Ministry of Food and Drug Safety (MFDS, South of Korea) tried to monitor contamination of organisms in confectionery products (such as biscuits, candies, chewing gums and ice candies) distributed in South Korea. MFDS evaluated the levels of indicator organisms: total aerobic bacteria, coliforms, Escherichia coli as well as the levels of food-borne illness organisms: Staphylococcus aureus, Bacillus cereus, Clostridium perfringens. Experimental plans for microbiological test were in accordance with the International Commission on Microbiological Specifications for Food (ICMSF). For this study, 1,005 samples were collected and from Seoul and Gyeongin region, South Korea. The average level of total aerobic bacteria in 1,005 samples was 1.7 log Colony Forming Unit(CFU)/g and the detection rate was 26.8%. The average level of Bacillus cereus was detected in 1.7 log CFU/g and the rate was 0.9%. There was no detection of coliforms, Escherichia coli, Staphylococcus aureus and Clostridium perfringens. The results of this study will be provided as the basic data to set the reasonable microbiological criteria of Korea Food Code.

Virulence Profile and Antimicrobial Resistance of Escherichia coli from Flies Captured from Agricultural Environment (농업환경에 서식하는 파리에서 분리된 E. coli의 병원성 유전자 및 항생제 내성 조사)

  • Yun, Bohyun;Jang, Youn Jung;Kim, Yeon Rok;Kim, Hwang-Yong;Kim, Won-Il;Han, Sanghyun;Kim, Se-Ri;Ryu, Jae-Gee;Kim, Hyun Ju
    • Journal of Food Hygiene and Safety
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    • v.32 no.2
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    • pp.147-153
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    • 2017
  • The purpose of this study was to isolate Escherichia coli from flies and to assess pathogenic genes and antibiotic resistance of the isolates. A total of 188 flies were captured in agricultural environment including fruits farms (n = 19), fermented soybean farms (n = 9), municipal waste (n = 46), livestock farms (n = 66), slaughterhouses (n = 38), and manure ground (n = 10). E. coli isolates of captured flies were tested for pathogenic gene and antibiotic resistance using PCR methods and VITEK2 systems. As a result, E. coli from 63% (119/188) of the captured flies has been detected, and the detection rate of E. coli was the highest (89%, 31/34) in flies captured at particular slaughterhouse. Of the 34 isolates, 94% (32/34) were pathogenic gene (ST gene) positive. Twenty-six percent (31/119) of the E. coli isolates were observed being resistant to one or more antibiotics. Markedly, one of E. coli isolates from Livestock farms was resistant to 7 antibiotics including ampicillin, ampicillin/sulbactam, cefazolin, cefotaxime, gentamicin, levofloxacin, and trimethoprim/sulfamethoxazole. In addition, it was ESBL positive. The results of the present study may suggest a risk of transmission of pathogenic and antimicrobial resistant bacteria from flies to livestock environment Therefore, it may need to prevent introducing flies into the agricultural production environment for safe food production.

A Report on the Shigella Cultures Isolated in Korea (1972) (1972년(年) 한국(韓國)에서 분리(分離)된 이질균(痢疾菌)에 관(關)한 보고(報告))

  • Ryu, Young-Hat;Kim, Soon-Hee
    • The Journal of the Korean Society for Microbiology
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    • v.8 no.1
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    • pp.7-11
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    • 1973
  • The authors identified eighty-eight Shigella cultures among about four thousands specimens collected from all over the country in 1972. Of eighty-eight cultures, seventy-seven cultures belonged to Shigella flexneri and eleven cultures to Shigella sonnei. None of cultures belonging to either subgroup A or C was detected in 1972. Of seventy-seven cultures of Shigella flexneri one was $B_{1b}$, fifty-six were $B_{2a}$, nine were $B_{3a}$, six were $B_{4a}$, three were By and one was each of $B_{3b}$ and $B_{3c}$. Of eleven cultures of Shigella sonnei seven cultures appeared to be phase I and the others phase II. Although there was quite a difference found in the incidence of isolating Shigella organisms between different areas as shown in Table 1, it would not be possible to understand that there might not have been the cases or carriers of Shigella in the areas where the organisms were not isolated in 1972. Concerning the biochemical properties it was not possible to compare the results obtained from the decarboxylase and dihydrolase tests with them obtained in previous years except that of lysine decarboxylase tests since they were not reported individually by the different serotypes in the previous reports. These results obtained in 1972 would be the data for the future comparison. In regards to the antibiotics-sensitivity of Shigella cultures the most of them showed sensitive results to nitrofurantoin, ampicillin, cephalosporin, gentamycin and geopen, and the majority of them appeared to be resistant to cloxacillin, tetracycline and streptomycin by means of the In Vitro tests.

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Gene analysis of galectin-1, innate immune response gene, in olive flounder Paralichthys olivaceus at different developmental stage (넙치, Paralichthys olivaceus 발생단계별 galectin-1 유전자의 발현 분석)

  • Jang, Min Seok;Lee, Young Mee;Yang, Hyun;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Hwang, In Joon;Kim, Sung Yeon
    • Journal of fish pathology
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    • v.26 no.3
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    • pp.255-263
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    • 2013
  • The innate immune response is fundamental defense response of vertebrates and invertebrates. Especially, the innate immune response important for larvae that lack of resistance to infectious diseases in the early stages. Galectin is one of the kinds of lectin and presents in the fish mucous that involves innate immune response. Galectin have been studied from various fishing species, but expression analysis of galectin is still unclear during early developmental stage in olive flounder. In this study, we investigated gene expression of galectin-1 from various developmental stage and tissues. We excised several tissues including the muscle, fin, eye, gill, brain, stomach, intestine, kidney, spleen and liver from adult olive flounder and confirmed gene expression of galectin-1 using RT-PCR and quantitative real-time PCR. Expression of galectin-1 was significantly higher in muscle, stomach and intestinal tissue than other tissue in adult fish (5 and 29 months). Also, galectin-1 gene was detected from 0 DAH and gradually increased to 35 DAH and since then decreased after stomach development period. Induction of galectin-1 during the early developmental stage suggest that muscle, fin and eye tissue is formed and begins the secretion of galectin this period. In addition, increased expression levels at 35 DAH suggest that due to complete formation of stomach and intestine, increase of secretion and activation of enzyme. This study shows that expression of galectin-1 during early developmental stages and adult period in olive flounder and can be expect that galectin-1 play essental role in the innate immune system throughout the whole life time. Galectin-1 is primary barrier such as skin and digestive tissue against pathogen infection, also digestive tract developmental period is important for pathogen invasion can be expected that it will serve. Mass mortality due to the disease in seed production is continuing damage, therefore these result will be meaningful about infectious disease during early developmental stages as a basic data for the study.