• Title/Summary/Keyword: fish detection

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A Lateral Flow Immunoassay Kit for Detecting Residues of Four Groups of Antibiotics in Farmed Fish (어류 중 4계열 잔류 항생물질 검출을 위한 Lateral Flow Immunoassay Kit 개발)

  • Jo, Mi Ra;Son, Kwang Tae;Kwon, Ji Young;Mok, Jong Soo;Park, Hong Jae;Kim, Hyun Yong;Kim, Gyung Dong;Kim, Ji Hoe;Lee, Tae Seek
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.48 no.2
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    • pp.158-167
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    • 2015
  • A lateral flow immunoassay kit based on antigen-antibody interactions was developed to detect residues of beta-lactams, quinolones, tetracyclines, and sulfonamides in farmed fish. Group-specific antibodies showing cross-reactivity with other antibiotics in the same group were produced in rabbits. The rabbits were immunized eight times to obtain the maximum titers. Antibodies were extracted from the antisera collected from the immunized rabbits and produced group-specific reactions with antibiotics from the four groups. A kit was prepared that optimize conditions for the antigen-antibody reaction, using colloidal gold conjugated antibodies, and was designed to detect the four groups of antibiotics simultaneously. The kit enabled the detection of antibiotics in the four groups at below maximum residue limits (MRLs), which were $200{\mu}g/kg$ for tetracyclines, $100{\mu}g/kg$ for sulfonamides, $50{\mu}g/kg$ for beta-lactams, and $100{\mu}g/kg$ for quinolones. The cross-reactivity of the antibodies ranged from 10-80% for the sulfonamides, 20-100% for tetracyclines, 38-100% for quinolones, and 20-100% for the beta-lactams, confirming that the antibodies were group specific. The test kit was used 30 times to examine spiked antibiotics at the limits of detection (LODs) and all produced positive results, indicating high sensitivity. The LODs for the assay ranged from 4-20 ng/mL for beta-lactams, 25-50 ng/mL for sulfonamides, 20-100 ng/mL for tetracyclines, and 30-80 ng/mL for quinolones, and there were no false negative reactions at above these LODs. In addition, all of the LODs of the developed kit were correlated with high-performance liquid chromatography (HPLC) data. Our lateral flow immunoassay kit can simultaneously detect antibiotic residues from a large number of fish samples rapidly, strengthening the safety of domestic farmed and imported fish.

Distribution of Pathogenic Vibrios and Environmental Factors Affecting Their Occurrence in the Seawater of Live Fish Tank (여름철 활어조 해수에서 병원성 비브리오균의 분포 및 환경인자와의 관계)

  • 김지희;박정흠;이태식;이희정;김성준
    • Journal of Food Hygiene and Safety
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    • v.16 no.3
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    • pp.241-246
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    • 2001
  • Distribution of pathogenic vibrios in the seawater of live fish tank and effect of environmental factors on their existence were investigated by collecting samples from fish markets and restaurants in 6 different cities. Pathogenic vibrios and coliforms were determined by using the most probable number (MPN) procedure, and aerobic plate count was enumerated by the standard pour plate method. No Vibrio chulerae O1 was detected in all the samples tested. Detection rates of V. cholerae non-O1, V. parahaemolyticus and V. vulnificus in all the smaples tested were 7.7%, 69.2% and 23.1%, respectively. Water temperature and trubidity of the seawater measured were higher in the pathogenic vibrios positive samples than in those negative samples. However, higher salinity and pH were shown in the pathogenic vibrios negative samples than in positive samples. The aerobic plate counts and MPN or total and focal coliforms in the seawater were higher in the presence of pathogenic vibrios than in the absence of pathogenic vibrios. In this study, the presence of pathogenic vibrios in the seawater tested was closely related with other physiochemical parameters and populations of coliforms, indicators far food safety.

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Omni-directional Vision SLAM using a Motion Estimation Method based on Fisheye Image (어안 이미지 기반의 움직임 추정 기법을 이용한 전방향 영상 SLAM)

  • Choi, Yun Won;Choi, Jeong Won;Dai, Yanyan;Lee, Suk Gyu
    • Journal of Institute of Control, Robotics and Systems
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    • v.20 no.8
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    • pp.868-874
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    • 2014
  • This paper proposes a novel mapping algorithm in Omni-directional Vision SLAM based on an obstacle's feature extraction using Lucas-Kanade Optical Flow motion detection and images obtained through fish-eye lenses mounted on robots. Omni-directional image sensors have distortion problems because they use a fish-eye lens or mirror, but it is possible in real time image processing for mobile robots because it measured all information around the robot at one time. In previous Omni-Directional Vision SLAM research, feature points in corrected fisheye images were used but the proposed algorithm corrected only the feature point of the obstacle. We obtained faster processing than previous systems through this process. The core of the proposed algorithm may be summarized as follows: First, we capture instantaneous $360^{\circ}$ panoramic images around a robot through fish-eye lenses which are mounted in the bottom direction. Second, we remove the feature points of the floor surface using a histogram filter, and label the candidates of the obstacle extracted. Third, we estimate the location of obstacles based on motion vectors using LKOF. Finally, it estimates the robot position using an Extended Kalman Filter based on the obstacle position obtained by LKOF and creates a map. We will confirm the reliability of the mapping algorithm using motion estimation based on fisheye images through the comparison between maps obtained using the proposed algorithm and real maps.

Detection of genetic abnormalities in human sperm, oocytes, and preimplantation embryos using fluorescence in situ hybridization (FISH) (Fluorescence in situ hybridization(FISH) 기법을 이용한 인간 생식세포 및 착상전 배아의 유전이상 검색)

  • 방명걸
    • Proceedings of the Korean Society of Developmental Biology Conference
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    • 1998.07a
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    • pp.12-18
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    • 1998
  • Tremendous progress has been made over the past quarter-century studying the genetics of gametogenesis and the resulting gametes and embryos. Studies merging molecular techniques and conventional cytogenetics are now beginning to bridge the gap between what we have learned about the meiotic process in males and females and what we know of the mitotic chromosomes of zygotes. Numerical abnormalities in sperm, oocytes and embryo can now diagnosed by fluorescence in situ hybridization (FISH). "At risk" couples can, therefore, have only unaffected embryos replaced in the sterus and avoid the possibility of terminating a pregnancy that might only be diagnosed as affected later gestation. Single-cell genetic analysis has also provided powerful tools for studying genetic defects arising during early human development. Recent studies of sperms, oocytes and cleavage-stage human embryos have revealed an unexpectedly high incidence. These genetic abnormalities are likely to contribute to early pregnancy loss and have important implications for improving pregnancy rates in infertile couples by assisted reproduction. The widespread use of preimplantation genetic diagnosis (PGD) awaits further documentatio of safety and accuracy. Other issues also must be addressed. First, the ethical issues regarding germ cell and embryo screening must be addressed including what diseases are serious enough to warrant the procedure. Another concern is the use of this technology for non-genetic disorders such as gender selection. Finally, the experimental nature of these procedure must continually be discussed with patients, and long-term follow-up studies must be undertaken. Development of more accurate and less expensive assays coupled with improved assisted reproductive technology success rates may make PGD a more widely use clinical tool. The future awaits these development.velopment.

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Detection of Marine Birnavirus(MABV) from Marine Fish in the Southern Coast of Korea (남해안 자연산 어류에서 Marine birnavirus(MABV)의 검출)

  • Yun, Hyun-Mi;Kim, Seok-Ryel;Lee, Wol-La;Jung, Sung-Ju;Oh, Myung-Joo
    • Journal of Aquaculture
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    • v.21 no.1
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    • pp.13-18
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    • 2008
  • Marine birnavirus(MABV) are well known fish pathogens in Asian countries such as Korea, Japan, and China. Prevalence of viral disease, geological distribution and host or reservoir of viruses were investigated from wild marine fishes in southern coast of Korea in 2003 and 2005. RT-PCR results showed that MABV were detected in 17 fishes(10.6%) from 160 fishes. Comparative analysis of the nucleotide sequences and the deduced amino acids of MABV genome from wild fishes were similar to reference strains of MABV and distinguished with IPNV strains.

Bacteriological and Chemical Hazard Analysis in Commercial Fish Products Minimally Processed (시판 어류 단순가공품의 세균학적 및 화학적 위해요소 분석)

  • Kim, Hyun-Jung;Lee, Dong-Soo;Kim, Il-Hoe;Kim, Young-Mog;Shin, Il-Shik
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.52 no.1
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    • pp.19-26
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    • 2019
  • The objective of this study was to analyze bacteriological and chemical hazards in minimally processed commercial fish products, including Hwangtae (freeze-dried pollock), dried anchovy, fermented anchovy sauce, and salted and dried yellow croaker. Escherichia coli counts from all samples were below the regulation limits of the Korean Ministry of Food and Drug Safety Standards on Quality of Seafood and Seafood Products (Food Code). However, the food poisoning bacterium Staphylococcus aureus was detected at levels above $1.0{\times}10^2$ colony forming units (CFU)/g in Hwangtae, dried anchovy, and salted and dried yellow croaker, which are commonly ingested without heating and pose bacteriological hazards. The detection of S. aureus, an organism indicative of poor personal hygiene, which can be introduced by employees and multiply during distribution, indicates the necessity of improving the sanitary control of minimally processed commercial fish products. Histamine was not detected from dried anchovy or salted and dried yellow croaker, but was detected at some of the highest levels in fermented anchovy sauces. This result suggests that efforts to reduce the amount of histamine in fermented anchovy sauces are required.

Monitoring and Development of Pretreatment Method for Perfluorinated Compounds in Fish Samples (어류에서의 과불화합물 분석을 위한 전처리법 확립 및 모니터링)

  • Heo, Jin-Ju;Lee, Ji-Woo;Kim, Seung-Kyu;Oh, Jeong-Eun
    • Journal of Korean Society of Environmental Engineers
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    • v.35 no.7
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    • pp.485-494
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    • 2013
  • An efficient extraction method was developed for the analysis of 16 perfluorinated compounds (PFCs) in fish samples. We compared the recoveries from the Ion-Pairing Extraction (IPE), Liquid-Liquid Extraction (LLE), Solid-Phase Extraction (SPE), IPE + SPE method with varying the injection time of the internal standard. As a result, IPE method with the internal standard before extraction was evaluated as the most effective pretreatment method. The RPM (Revolution Per Munite) and pH in IPE-before method were additionally adjusted and the more efficient pretreatment method was established. The total 33 fish samples including liver and gut samples were collected from Korean markets and analyzed PFCs with developed pretreatment method of this study. Total 16 PFC levels in fish samples ranged from ND to 1.67 ng/g with 100% detection frequency. The average PFCs concentrations of muscle, liver and gut samples from fish were compared and showed the following trend: liver (17.8 ng/g) > gut (13.3 ng/g) > muscle (1.67 ng/g). The PFC levels in fish samples were similar or lower than other available previous results of foreign studies.

Histopathology and residues in fresh water fish exposed to acute and chronic copper and mercury toxicity

  • Sawsan, H.A.;Amira, H.M.;Mostafa, M.B.;Nashaat, AM.M.
    • Journal of fish pathology
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    • v.30 no.2
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    • pp.115-134
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    • 2017
  • A total number of 668 apparently healthy fish were obtained from farm to study the effect of two heavy metals (Copper and Mercury) on histopathology of liver, kidney, spleen, gills and muscles also residues in muscles. The $LC_{50}$/96 hr. of Cu and Hg were estimated and fish exposed to 1/2 $LC_{50}$ for 7 days and for 1/10 $LC_{50}$ for 8 weeks from each product separately. Histopathological findings in acute and chronic mercuric chloride toxicity revealed degeneration and necrosis in the glomeruli, interstitium tissue and epithelium lining renal tubules. The tubular epithelium became necrotic at several places. Eosinophilic hyaline droplets is exist in the cytoplasm of the necrosed cells. Degenerative changes and hyperactivity in melanomachrophage center was seen in the spleen together with some necrotic areas. Necrosis and aggregation of melanomachrophage were seen in the hepatic cells, Hepatic cells showed vacuolar degeneration in the hepatic cells. Gills showed loss in the lamellae of the filaments associated with edema, inflammatory cells infiltration and haemorrhages in the arch. The sarcoplasm of the bundles of the skeletal muscle showed granular degeneration and focal inflammatory cells infiltration between the hyalinized bundles. Mercury residues obtained from these studies in the acute toxicity were 0.22 ppm/gm in the 2nd day, 0.411 ppm/gm in the $5^{th}$ day ended with 0.96 ppm/gm in the $7^{th}$ day. In chronic toxicity it was 1.1320, 1.7140, 2.3620 and 3.5640 ppm/gm respectively from the $2^{nd}$ to the $8^{th}$ week of exposure. In acute and chronic copper toxicity, there was degenerative changes in renal tubules. Melanophores aggregation in the wall of the blood vessels of the spleen and depletion of some of the melanophores in the melanomachrophage were seen together with necrosis in some areas. Congested Mvs (Micro vessels) and vacuolation of hepatocytes were observed. Some areas of hemorrhage and melanophores vacuolar degeneration in the liver were seen. There was mitosis in some areas with displesia of hepatopancreatic cells and eosinophilic granular cells aggregation. Zymogen granules disappeared and there were dyplastic hepatocytes. Congestion in the blood vessels of the gill filaments, associated with massive number of granular eosinophilic cells infiltration were seen in the base of the filaments. There were sever vacuolization and hyalinization in the skeletal muscle bundles. Detection of residues of copper sulfate revealed increase of the amount of copper measured in ppm/gm comparing to the normal control starting from 0.60 ppm/g in the $2^{nd}$ day, 0.67 ppm/g in the $5^{th}$ day and 0.67 ppm/g in the $7^{th}$ day. Result obtained in chronic copper sulfate toxicity revealed gradual increase of the amount of copper which ranged from 0.18 ppm/g at the $2^{nd}$ week to 0.21 ppm/g in the $8^{th}$ week of exposure.

Analysis of Chromosome aberrations by fluorescence in situ hybridization using triple chromosome-specific probes in human lymphocyte exposed to radiation (3중 DNA probe를 이용한 FISH(fluorescence in situ hybridization) 기법으로 방사선에 의한 염색체 이상 분석)

  • Chung, Hai-Won;Kim, Su-Young;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.24 no.1
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    • pp.45-53
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    • 1999
  • Fluorescence in situ hybridization with chromosome-specific probe has been shown to be a valid and rapid method for detection of chromosome rearrangements induced by radiation. This method is useful for quantifying structural aberrations, expecially for stable ones, such as translocation and insertion, which are difficult to detect with conventional method in human lymphocyte. In order to apply FISH method for high dose biological dosimetry, chromosomal abberations by radiation at doses of 1, 3, 5, and 7Gy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. The frequencies of stable translocation per cell equivalent were 0.04, 0.33, 1.22, 2.62, and 5.58 for the lymphocyte exposed to 0, 1, 3, 5, and 7Gy, respectively, and those of dicentric were 0.00, 0.06, 0.52, 1.19 and 2.44, respectively. Significantly more translocation of t(Ab), a translocated chromosome with a piece of painted acentric matrial 'b' attached to unpainted piece containing centromere 'A', than reciprocal chromosome t(Ba) was observed. The frequencies of all type of chromosome rearrangements increased with dose. From above result, FISH seemed to be useful for radiation biodosimetry by which the frequencies of various types of stable aberrations in human lymphocyte can be observed more easily than by conventional method and so will improve our ability to perform meaningful biodosimetry.

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Retrospective Study of ALK Rearrangement and Clinicopathological Implications in Completely Resected Non-small Cell Lung Cancer Patients in Northern Thailand: Role of Screening with D5F3 Antibodies

  • Tantraworasin, Apichat;Lertprasertsuke, Nirush;Kongkarnka, Sarawut;Euathrongchit, Juntima;Wannasopha, Yutthaphan;Saeteng, Somcharoen
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.7
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    • pp.3057-3063
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    • 2014
  • Background: Anaplastic lymphoma kinase (ALK) gene rearrangement in non-small cell lung cancer (NSCLC) has been intensively studied. The gold standard for ALK detection is FISH, but this is not routinely conducted in clinical practice, so that the IHC method has a role. The aim of this study was to identify the incidence of ALK rearrangement and risk or prognostic factors for ALK positivity using both of IHC and FISH methods. Materials and Methods: From January 2008 to December 2012, 267 completely resected NSCLC patients in Chiang Mai University Hospital were enrolled in this study. Clinical and pathological variables and outcomes of treatment were retrospectively reviewed. IHC and FISH were used to evaluate ALK rearrangement. Sensitivity and specificity of IHC were analyzed. Multivariable analysis was used to identify clinico-pathological correlations with positive results of IHC and clinical outcomes. Results: Twenty-two (8.2%) of 267 specimens were IHC-positive for ALK with intense cytoplasmic staining, whereas only 10 (3.8%) were FISH-positive. Sensitivity, specificity and the positive likelihood ratio with IHC were 80.0%, 94.9%, and 15.8 respectively. Age less than 55 years (RR 4.4, 95%CI 1.78-10.73, p value=0.001) and presence of visceral pleural invasion (VPI) (RR 2.9, 95%CI 1.21-6.78, p value =0.017) were identified as risk factors for ALK rearrangement with FISH. There were no statistically significant differences in other clinical and pathological variables. ALK rearrangement was not a prognostic factor for tumor recurrence or overall survival. Conclusions: The incidences of ALK positivity in completely resected NSCLCs in northern Thailand were 8.2% by IHC and 3.8% by FISH. IHC with mouse monoclonal, Ventana D5F3 antibody can be used as a screening tool before FISH method because of high specificity and high positive likelihood ratio. Age less than 55 years and VPI are risk factors for ALK positivity.