• Title/Summary/Keyword: Species detection

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Monitoring of Malachite Green in Freshwater Fish using LC-MS/MS (LC-MS/MS를 이용한 담수 어류 중 말라카이트 그린 분석)

  • Choi, Hee-jin;Yuk, Dong-Hyun;Park, Young-Ae;Jung, Bo-Kyeng;Hong, Mi-Sun;Yoon, Yong-Tae;Yi, Hye-Jin;Kim, Youn-Cheon;Park, Sung-Kyu;Kim, Moo-Sang;Jung, Kweon
    • Journal of Food Hygiene and Safety
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    • v.31 no.1
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    • pp.15-20
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    • 2016
  • Malachite green was measured in 200 freshwater fish collected from local markets in Seoul using HPLC-DAD and LC-MS/MS. LC-MS/MS method was validated by linearity, accuracy, precision and limits of detection and quantification according to the CODEX's recommendation and HPLC-DAD method was applied according to the Food Code. Malachite green levels above the quantification limit of the LC-MS/MS were determined 18.5% (37) but just 1 fish was shown to contain malachite green by HPLC-DAD. Of 83 domestic fish, 21 fish were detected malachite green (25.3%). Of 117 fish from China, just 16 fish were detected malachite green (13.4%). In detection rate by species carp (35.0%), Crucian carp (30.4%), cat fish (28.0%), Korean bull head (23.8%), snake head (20.0%), eel (10.5%) and loach (7.8%) were in order. Especially, fish collected at summer were shown to contain malachite green frequently; the detection rate was 54.8%.

Evaluation of Peptide Nucleic Acid Probe-Based Fluorescence In Situ Hybridization for the Detection of Mycobacterium tuberculosis Complex and Nontuberculous Mycobacteria in Clinical Respiratory Specimens (임상 객담검체에서 Peptide Nucleic Acid Probe를 이용한 결핵과 비결핵 항산균의 구분)

  • Lee, Seung Hee;Kim, Shine Young;Kim, Hyung Hoi;Lee, Eun Yup;Chang, Chulhun L.
    • Annals of Clinical Microbiology
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    • v.18 no.2
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    • pp.37-43
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    • 2015
  • Background: Tuberculosis is globally the most important cause of death from single pathogen. Rapid and accurate identification of mycobacteria is essential for the control of tuberculosis. We evaluated a fluorescence in situ hybridization (FISH) method using peptide nucleic acid (PNA) probes for the differentiation of Mycobacterium tuberculosis complex (MTB) and nontuberculous mycobacteria (NTM) in direct smears of sputum specimens. Methods: The cross-reactivity of MTB- and NTM-specific PNA probes was examined with reference strains of M. tuberculosis ATCC 13950, Mycobacterium kansasii ATCC 12479, Mycobacterium fortuitum ATCC 6841, several clinical isolates of mycobacteria (Mycobacterium abscessus, Mycobacterium avium, Mycobacterium intracellulare, Mycobacterium gordonae and Mycobacterium chelonae), and 11 frequently isolated respiratory bacterial species other than mycobacteria. A series of 128 sputa (89 MTB culture positive, 29 NTM culture positive, and 10 under treatment culture negative) with grades of trace to 4+ were used to evaluate the performance of the method. Results: The MTB- and NTM-specific PNA probes showed specific reactions with the reference strains of MTB and M. kansasii and clinical isolates of mycobacteria except M. fortuitum ATCC 6841, and no cross-reactivity with other tested bacteria. The PNA probe-based FISH assay for detection of MTB had a sensitivity and specificity of 100%, respectively. The sensitivity and specificity of the NTM-specific PNA probe was 100%. The smear grades of the PNA FISH test were same as with those of the fluorescence AFB stain in 2+ or higher grade. Conclusion: Detection and differentiation based on PNA FISH is sensitive and accurate for detecting mycobacteria and for differentiating MTB from NTM in clinical sputum smears.

Investigation of Microbial Contamination in the Raw Materials of Meal Kits (간편조리세트 원재료의 미생물 오염도 조사)

  • Hyun-Kyung Lee;Young-Sook Do;Min-Jung Park;Kyoung Suk Lim;Seo-In Oh;Jeong-Hwa Lim;Hyun-Soo Kim;Hyun-Kyung Ham;Yeo-Jung Kim;Myung-Jin Lee;Yong-Bae Park
    • Journal of Food Hygiene and Safety
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    • v.39 no.2
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    • pp.109-117
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    • 2024
  • This study investigated the microbial contamination of agricultural, livestock, and marine ingredients in 55 meal kits distributed across Gyeonggi-do, South Korea. Of the 55 meal kits, 48 contained agricultural ingredients, 43 contained livestock ingredients, and 16 contained marine ingredients. The detection rate of the total aerobic bacteria in the agricultural, livestock, and marine products was 100%. The average numbers of the total aerobic bacteria were 6.57 log colony-forming units (CFU)/g in the agricultural products, 4.60 log CFU/g in the livestock products, and 5.47 log CFU/g in the marine products. The coliform detection rates in the agricultural, livestock, and marine products were 81.25%, 69.77%, and 43.75%, respectively. The average numbers of coliforms were 2.83 log CFU/g in the agricultural products, 1.34 log CFU/g in the livestock products, and 1.12 log CFU/g in the marine products. Escherichia coli was detected in 13 livestock products (30.23%), with levels ranging from 0.70 to 2.36 log CFU/g. Contrastingly, E. coli was detected in only one marine product (6.25%) and was not detected in any agricultural products. The detection rates of fungi in agricultural, livestock, and marine products were 97.92%, 93.02%, and 93.75%, respectively. The average numbers of fungi were 3.82 log CFU/g for the agricultural products, 2.92 log CFU/g for the livestock products, and 2.82 log CFU/g for the marine products. The isolation rates of foodborne pathogens from the agricultural, livestock, and marine products were 35.42%, 37.21%, and 31.25%, respectively. Forty-five foodborne pathogens of seven species, including Bacillus cereus and Salmonella spp., were isolated from the raw materials of the agricultural, livestock, and marine products in 55 meal kits. To prevent foodborne diseases caused by meal kits, it is necessary to focus on washing, heating, and preventing cross-contamination during cooking.

The Effects of Milking Time and Dairy Processes on Melatonin Contents in Milk (착유 시간 및 유처리 공정이 우유 내 멜라토닌 함량에 미치는 영향)

  • Kang, S.H.;Baick, S.C.
    • Journal of Dairy Science and Biotechnology
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    • v.26 no.1
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    • pp.1-4
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    • 2008
  • Melatonin is one of the functional hormones in the milk and found in all mammalian species. The primary motivation for the use of melatonin as a supplement is as a natural aid to better sleep. Melatonin contents in milk vary according to the naturally occurring factors such as seasons and milking times. As a number of studies indicate melatonin supplementation helps to reduce the age-related decline in hormone production and overall enhancement of health, the more detailed studies are required to know the nature of milk derived hormones and to clarify them as a functional value adaptation. The objectives of this study were to examine the change of melatonin content in bovine milk during different milking times at the two dairy farms and during dairy process including homogenization and pasteurization. Commercial dairy products including night time milking products(night milk) marketed in Japan were also investigated. Melatonin content was determined by radioimmunoassay using 125I. Individual milk was collected from lactating Holstein cows at Kyong-Ki province in Korea. At farm A, the melatonin contents milking at 4 a.m. was higher than those of 7 p.m. and 12 p.m.($6.90{\pm}3.55,\;2.01{\pm}1.47$ and $0.16{\pm}0.04pg/mL$, respectively). At farm B, the mean melatonin contents milked from 24:00 AM to 05:00 AM have shown the highest contents ($4.65{\pm}$0.72pg/mL), and milking samples of the rest time had very little or non-detection of melatonin contents. Melatonin contents of commercial night milk samples were about 5 times higher than those of other common milk products. These results suggested that melatonin might be heat-resistant and commercial dairy process might not affect its contents in the final products.

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Study of Bacteria Associated with Dental Caries Using a 3 Tone Disclosing Agent (세가지 색상차이를 보이는 착색제를 이용한 치아 우식 관련 균에 관한 연구)

  • Lee, Jeongeun;Park, Howon;Lee, Juhyun;Seo, Hyunwoo;Lee, Siyoung
    • Journal of the korean academy of Pediatric Dentistry
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    • v.45 no.1
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    • pp.32-40
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    • 2018
  • The aim of this study was to determine the efficacy of a 3 tone plaque disclosing gel in assessing the risk of caries related to the population of Streptococcus mutans, Streptococcus sobrinus, and Lactobacillus spp. quantified using a quantitative real-time polymerase chain reaction (qRT-PCR). 15 healthy children of ages 9 - 12 years were randomly examined. The 3 tone plaque disclosing gel was applied on teeth surfaces, which changed the color to pink or red, blue or purple and light blue. Plaque was divided into 3 groups based on staining. Genomic DNA from each sample was subjected to a qRT-PCR assay for quantitative detection of target bacteria. The Kruskal-Wallis test was conducted for correlation between the color of plaque and the number of bacterial species. The levels of S. mutans, S. sobrinus, and Lactobacillus spp. were significantly different in the plaque samples of the 3 groups (p < 0.05). The proportion of S. sobrinus to S. mutans showed correlation to the color of plaque. The different color-dyed plaque was related to the number of acidogenic bacteria. The 3 tone plaque disclosing gel could be used as one of the indicators to assess the clinical risk of caries associated with the population of S. mutans, S. sobrinus, and Lactobacillus spp.

Cloning and High Expression of Nattokinase Gene from Bacillus subtilis BB-1 (Bacillus subtilis BB-1으로부터 나토키나아제 유전자 크로닝 및 대량발현)

  • Lee Young-Hoon;Lee Sung-Ho;Park Ki-Hoon;Choi Young-Ju;Jeong Yong-Kee;Gal Sang-Wan
    • Journal of Life Science
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    • v.16 no.2 s.75
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    • pp.274-281
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    • 2006
  • A fibrinolytic enzyme gene was isolated from Bacillus subtilis BB-1 by PCR method. Primers for PCR cloning were designed according to pre-identified gene for fibrinolytic enzymes from B. subtilis. The primer sequences were 5'-CGG ATC CGT GAG AGG CAA AAA GGT G-3' and 5'-TGA ATT CTT AAT GTG CTG CTG CTT GTC C-3' as concensus sequences of the fibrinolytic genes of Bacillus species. The PCR product was 1,145 bp and the sequence homology was 99% with nattokinase gene isolated from Japanese natto. The cloned fibrinolytic gene was reconstructed in Bacillus-E. coli shuttle vector, pEB for bulk-production. The fibrinolytic enzyme was purified by FPLC from the cloned B. subtilis 168. The optimum pH and temperature of the enzyme were 7.0 and $35^{\circ}C$, respectively. The fibrinolytic enzyme did not show any activity toward to skim milk, gelatin, casein and blood agar plate. The enzyme specific polyclonal antibody was prepared in rabbit for further assays such as detection of the gene expression in plant cells. This means that the enzyme may be used for health-care such as thrombosis without any hamful effects in the blood vessel.

Detection of a Quorum-Sensing Inhibitor from the Natural Products (천연물로부터 Quorum Sensing 저해제의 탐색)

  • Kim, Tae-Woo;Cha, Ji-Young;Lee, Jun-Seung;Min, Bok-Kee;Baik, Hyung-Suk
    • Journal of Life Science
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    • v.18 no.2
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    • pp.206-212
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    • 2008
  • The quorum sensing (QS) regulatory network has been the subject of extensive studies during recent years and has also attracted a lot of attention because it both positively and negatively regulates various putative virulence factors, although initially considered to be a specialized system of Vibrio fischeri and related species. In this study, to identify the novel materials which inhibit QS system of microorganisms, extracts of eighteen natural products were tested by bioassay using N-(3-oxohexanoyl)-$_L$-homoserine lactone and N-(3-oxooctanoyl)-$_L$-homoserine lactone synthesized in this experiment and an Agrobacterium tumefaciens NT1 biosensor strain containing a traI::lacZ fusion. The result indicated that the extracts of cabbage, leek, and onion exhibited the QS inhibition activity. Thus, materials contained in the extracts were isolated via recycling preparative HPLC and were purified via a JAIGEL-LS255 column. The common fraction corresponding to a peak of the 83 min point of them quenched the quorum sensing of A. tumefaciens NT1 biosensor strain in ABMM containing X-gal and was designated quorum sensing inhibitor-83 min (QSI-83). The QSI-83 exhibited the heat stability and did not inhibit the growth of A. tumefaciens NTl. Furthermore, thin layer chromatography (TLC) results suggested that these novel materials may be antagonists of N-acyl homoserine lactone or may inhibit the QS autoinducer synthesis by Pseudomonas syringae pv. tabaci.

Ecotoxicity Assessment of Leachate from Disposal Site for Foot-and-Mouth Disease Carcasses (구제역 가축 매몰지 침출수 독성영향평가)

  • Kim, Dongwoo;Yu, Seungho;Chang, Soonwoong;Lee, Junga
    • Journal of the Korean GEO-environmental Society
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    • v.15 no.8
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    • pp.5-11
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    • 2014
  • In this study, chemical analysis and ecotoxicity tests of leachate from disposal site for foot-and-mouth disease carcasses (FMD leachate) were conducted to collect fundamental data that will be used to develop environmental risk assessment tools for FMD leachate. For chemical analysis, concentration of $Cl^-$, $NH{_4}{^+}-N$, Korea standard method indicators for detection of leachate released from animal carcasses burial site into groundwater and NRN (Ninhydrin-Reactive Nitrogens), a newly suggested screening test indicator to detect groundwater contamination by FMD leachate, were assessed. For ecotoxicity tests, luminescent bacteria (V. fischeri), micro-algae (P. subcapitata) and water flea (D. magna) were selected as test species. Correlation analysis between the concentration of $Cl^-$, $NH{_4}{^+}-N$, NRN and the toxicity to V. fischeri was performed to identify the better indicators to monitor FMD leachate contamination. From regression analysis, the concentration of the indicators in FMD leachate contaminated sample that induced halfmaximal toxic effect to V. fischeri was evaluated. Results obtained from this study can be applied to assess the risk by FMD leachate and to establish the guideline to manage risk in relation to FMD leachate.

DETECTION OF SALIVARY STREPTOCOCCUS MUTANS LEVELS USING MONOCLONAL ANTIBODIES (단클론항체를 이용한 타액 내 Streptococcus mutans 수준의 측정)

  • Kim, Chu-Sung;Kim, Jae-Gon;Yang, Yeon-Mi;Baik, Byeong-Ju;Lee, Kyung-Yeol;Kim, Mi-Ah;Lim, Su-Min
    • Journal of the korean academy of Pediatric Dentistry
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    • v.37 no.2
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    • pp.186-192
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    • 2010
  • Streptococcus mutans, one of the major causal agents of dental caries, is component of the dental plaque. It produces various organic acids such as lactic acid which is the end-product of glycolysis, and this leads to dental caries. A new system using species-specific monoclonal antibodies was developed to detect Streptococcus mutans in saliva. The system quickly detects salivary Streptococcus mutans in 30min and classifies the result into two levels. The purpose of this study was to investigate correlation between monoclonal antibody-based detecting system and selective medium-based detecting methods. Children's deft indices were also compared with Streptococcus mutans counts in MSB agar plate. Subjects consisted of 15 children in the age of 3 to 6 years. They were assigned to three groups : Group I(deft index = 3), Group II(deft index $\leqq$3), Group III(deft index $\geqq$4). The results are as follows : 1. The rate of children with positive response was 13.3% and with negative response was 86.7% in the result of Saliva-checkTM Mutans test kit. 2. There was a positive correlation between monoclonal antibody-based detecting system and selective medium-based detecting methods(p<0.05). 3. Streptococcus mutans counts in MSB agar plate were irrelevant to deft of children(p=0.34).

Detection of Tissue-specific Expression of Porcine Cytochrome P450 Aromatase Genes by Use of Denaturing High Performance Liquid Chromatography(DHPLC) Technique (DHPLC 기술을 이용한 돼지 Cytochrome P450 Aromatase 유전자의 조직 - 특이적 발현양상 관찰)

  • Chae, S.H.;Ghlmeray, A.K.;Hong, J.M.;Lee, E.J.;Chang, J.S.;Choi, I
    • Journal of Animal Science and Technology
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    • v.46 no.3
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    • pp.315-324
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    • 2004
  • Cytochrome P450 aromatase is the enzyme responsible for biosynthesis of female sex hormone(estrogen) and 19-nortestosterone(nandrolone), a unique steroid hormone endogenously synthesized in the pig. By use of RT-PCR coupled with DHPLC technique (WAVE analysis), expression pattern of isoforms of porcine cytochrome P450 aromatase gene was investigated. Relatively higher expression of aromatase mRNA was observed in testis than in ovary and this result accounted for the previous findings of higher blood estrogen level in male compared with female in this species. The result from the DHPLC demonstrated that PCR amplified DNA fragments of ovary and testis tissues. using unique PCR primers for all three types of aromatase genes, were different from those of type II and ill genes. Further nucleotide sequence analyses of the plasmid clones containing the PCR products revealed that nucleotide sequences of all clones were identical to type I aromatase gene(ovary type). Thus, the result from the present study indicates that the ovary and testis express the same type of aromatase gene. Therefore, the efficacy of DHPLC techniques used for this study helped us to analyze tissue-specific expression of isoform of genes containing the nucleotide sequences with high homology.