• Title, Summary, Keyword: Zearalenone

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Development of Enzyme Linked Immunosorbent Assay for Determination of Zearalenone in Animal Feeds (가축사료 중 Zearalenone 분석을 위한 Enzyme Linked Immunosorbent Assay법의 개발)

  • 하정기;정덕화;김성영
    • Journal of Food Hygiene and Safety
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    • v.6 no.3
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    • pp.111-117
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    • 1991
  • We examined to develop the enzyme linked immunosorbent assay (ELISA) for determination of zearalenone in animal feeds. Zearalenone was first converted to 6'-(carboxymethyl) zearalenone oxime(zearalenone oxime) to get a coupling site and then conjugated to bovine serum albumin(BSA) for use as immunogen and to horseradish peroxidase(HRP) for use as enzyme marker. Antibody against zearalenone was obtained after 11 weeks of immunization of rabbit with zearalenone oxime-BSA. Cross reactivity of the antibody with ${\alpha}-zearalanol,\;{\beta}-zearalenol,\;{\alpha}-zearalanol\;and\;{\beta}-zearalanol$ were 168, 46, 26 and 20% respectiviely. A simple procedure was devised for the screening of zearalenone in feeds using ELISA. Feeds samples(5g) were extracted by blending with 25 ml of methanol-phospate butTered saline-dimethylformate(70 : 29 : 1) and the extract was filtered and aqueous filterate analyzed. It took only 1 hours to do whole procedure for the analysis of zearalenone in feeds by the direct competitive ELISA, and detectable limit was 1-100 ppb. Using this procedure, only 4 of 24 feed samples showed positive results with 3.93-7.43 ppb levels.

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Establishment of Indirect Competitive ELISA for the Detection of Zearalenone Produced by Fusarium sp. (Fusarium속이 생성하는 zearalenone 측정을 위한 Indirect Competitive ELISA의 확립)

  • 강성조;정덕화
    • Journal of Food Hygiene and Safety
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    • v.13 no.4
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    • pp.419-424
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    • 1998
  • An enzyme-linked immunosorbent assay (ELISA) was established for the detection of zearalenone by using monoclonal antibodies produced by Z-M-26 hybridoma cells when injected into a mouse and zearalenone-oxime-OV A conjugate. Zearalenone-oxime-OV A conjugates were diluted with carbonyl buffer, coated to 96 well microtiter plates at $4^{\circ}C$ overnight and blocked with 1% BSA overnight. One thousand times diluted antibody solution together with standard zearalenone or sample was added to 96-well microtiter plates and stood overnight. A secondary antibody conjugated with HRP was added and an hour later, enzyme substrate (TMBZ) solution was added for color develpment. Mter 30 minutes, coloring reaction was terminated by adding 2 N $H_2S0_4$ and the O.D. was measured at 450 nm. Detection range of this method was about 0.1~100 ppb. The established indirect competitive ELISA method was suitable for a rapid and effective analysis of zearalenone in agricultural products.

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Production of Monoclonal Antibody against Zearalenone Produced by Fusarium sp. (Fusarium sp.가 생성하는 zearalenone에 대한 단크론성 항체생산)

  • Kang, Sung-Jo;Chung, Duck-Hwa;Kang, Jin-Soon
    • Korean Journal of Food Science and Technology
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    • v.30 no.6
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    • pp.1409-1414
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    • 1998
  • To develop zearalenone-specific monoclonal antibodies, hybridoma cells were produced by fusion of myeloma cells $(P3{\times}63Ag\;V653)$ and spleen cells from BALB/c female mice immunized with zearalenone-oxime coupled to bovine serum albumin (BSA). After screening of antibody titer of them with a sandwich type enzyme-linked immunosorbent assay (ELISA), 5 hybridomas which could produced monoclonal antibodies with a high affinity for zearalenone were selected. The monoclonal antibody produced by Z-2-M26 hybridoma exhibited the high sensitivity to zearalenone and a little cross-reactivity to ${\alpha}-zearalenol$ (11%), but did not react with ${\beta}-zearalenol,\;{\alpha}-zearalenol,\;{\beta}-zearalenol$ and DON. In conclusion, the developed monoclonal antibody appeared to be a very promising immunoreagent for the future development of a specific and sensitive quantitative ELISA for zearalenone.

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Studies on practical application of zearalenone ELISA kits (Zearalenone ELISA kits의 응용에 관한 연구)

  • Yoon, Hwa-joong;Kim, Tae-Jong;Lee, Sung-Yun;JeGal, Jun;Yoon, Ji-Byung
    • Korean Journal of Veterinary Research
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    • v.38 no.2
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    • pp.297-303
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    • 1998
  • For the extraction and measurement of zearalenone in the corn, bean, wheat and barley contaminated with Fusarium graminearum, the zearalenone-oxime, zearalenone-oxime BSA and zearalenone monoclonal antibodies were studied to develop and apply the direct competitive enzyme linked immunosorbent assay (ELISA). The extraction range of zearalenone with the monoclonal antibodies produced in this experiment was 10ng to 500ng/g feed and the 50% inhibition value was 50ng/ml. The mean recoveries of zearalenone artificially spiked in the ground corn were 89%. The specificity of F-2 monoclonal antibody for the analogues was favorable for the direct competitive ELISA. The result of the experiment showed the zearalenone in the corn, bean, wheat and barely naturally contaminated with the mold would be suitable for extraction and measurement with the monoclonal antibodies.

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Antifungal Activities of Equisetin, Zearalenone, and 8'-Hydroxyaearalenone Isolated from Fusarium Species against Plant Pathogenic Fungi. (Fusarium속 균주로부터 분리한 Equisetin, Zearalenone 및 8'-Hydroxyzearalenone의 식물병원곰팡이에 대한 항균활성)

  • 김진철;박중협;최경자;김흥태;최용호;조광연
    • Microbiology and Biotechnology Letters
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    • v.30 no.4
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    • pp.339-345
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    • 2002
  • Antifungal substances were isolated from solid cultures of Fusarium equiseti FO-68 obtained from arrowhead and Fusarium sp. FO-510 obtained from egg plant, and then their antifungal activities were investigated against plant pathogenic fungi in vitro and in vivo. An antifungal substance was purifed from rice solid cultures of F. equiseti FO-68 and identified as equisetin. In addition, two antibiotic substances were isolated from solid cultures of Fusarium sp. FO-510 and their chemical structures were determined to be zearalenone and 8'-hydroxyzearalenone. in vitro, equisetin and zearalenone inhibited mycelial growth of most of the plant pathogenic fungi tested, whereas 8'-hydroxyzearalenone hardly inhibited fungal growth. In vitro, equisetin effectively controlled the development of tomato gray mold and tomato late blight. Zearalenone exhibited in vivo antifungal activity against rice blast, rice sheath blight, tomato gray mold, and tomato late blight. However, 8'-hydroxyzearale-none did not control the development of plant diseases except tomato gray mold. This is the first report on the antifungal activities of equisetin, zearalenone, and 8'-hydroxyzearalenone.

Screening of zearalenone-producing strains by ELISA method (ELISA법에 의한 zearalenone 생성균주의 검색)

  • Kim, Sung-Young;Chung, Sun-Hee;Chung, Duck-Hwa
    • Applied Biological Chemistry
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    • v.36 no.1
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    • pp.7-10
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    • 1993
  • ELISA method was applied for the screening of zearalenone producing strains. The developed ELISA was as follow: $125\;{\mu}l$ of diluted solution (1 : 500) of antibody was added to each microtiter well and incubated overnight at $40^{\circ}C$. For direct competitive ELISA, samples and zearalenone-peroxidase conjugate were mixed in a 1 : 1 ratio, and a $100\;{\mu}l$ of aliquot was then added to antisera-coated wells. Plates were incubated for 30 minutes at $37^{\circ}C$, and wells washed 6 times, and $100\;{\mu}l$ of ABTS substrates was added. Plates were incubated for antother 15 minutes at $37^{\circ}C$, and $100\;{\mu}l$ of stopping reagent was added to the wells and absorbance was recorded at 410nm on ELISA Reader. Among 19 strains showed zearalenone-producing ability by ELISA, 3 strains (R-5, C-46, S-134) produced more than 50 ng/ml of zearalenone.

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Isolation of the Zearalenone-Producing Strains from Agricultural Products in Youngnam Districts (영남지방 농산물에서의 Zearalenone 생성균의 분리)

  • Kim, Sung-Young;Chung, Sun-Hee;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.24 no.6
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    • pp.581-585
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    • 1992
  • To isolate zearalenone-producing strains from agricultural products in Youngnam districts, various samples such as rice(60), barley(52), soybean(45), peanut(33), corn(32), maeju(27), unhulled rice(UR, 40), unhulled barley(UB, 54), and soil(32) collected. From 375 samples, 302 Fusarium strains were isolated. The isolated strains were cultured for 14 days at $30^{\circ}C$ in rice medium, and then screened zearalenone-producing strains by TLC and HPLC. The result was that 29 isolates[rice(3), maeju(2), corn(1), barley(5), soil(4), peanut(3), soybean(4), U.B.(5), U.R.(2)] were screened as zearalenone-producing strains by TLC, while 19 isolates [rice(2), maeju(1), corn(1), barley(4), soil(3), soybean(2), U.B.(3), U.R.(3)] by HPLC method.

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A Study on the Simultaneous Determination of Residual Zeranol, Zearalenone and Their Metabolites in Beef by Gas Chromatography/Mass Spectrometry (Gas Chromatography/Mass Spectrometry에 의한 우육 중의 잔류 Zeranol, Zearalenone 및 그 대사산물들의 동시 분석법에 대한 연구)

  • 이은섭;이용욱
    • Journal of Food Hygiene and Safety
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    • v.9 no.1
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    • pp.1-13
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    • 1994
  • A Simultaneous determination method was improved for the determination and confirmation of zeranol, zearalenone, as well as their isomers and metabolites, in beef. The analytes were extracted from tissue by CH3CN, hydrolyzed enzymatically(for glucuronide conjugates), cleaned up by a strong basic anion exchange resin combined with a liquid/liquid partitioning, derivatized using MSTFA and confirmed, quantified by GC/MS/SIM with a internal standard, zearalane. The results were as follows : (1) all the estrogens were separated on the GC/MS chromatogram under the extraction method and the chromatographic conditions improved, the retention times of zearalane-TMS2, zearalanone-TMS2, zearalenone-TMS2, zeranol-TMS3, taleranol-TMS3, and $\alpha$-zearalenol-TMS3, $\beta$-zearalenol-TMS3, were 18.49, 19.44, 19.63, 19.71, 19.79 and 19.99, 20.08 minutes, respectively. (2) The calibration curves of residual zeranol, zearalenone and their metabolites showed constantly linear(r=0.99) in the range of 5~20 ng. The minimum detection concentration of residual zeranol, zearalenone and their metabolites was 1 ppb. (3) The total average recovery of residual zeranol, zearalenone and their metabolites from spiked beef was 60.2%(CV=29.7%) at the 1 ppb and 63.5%(CV=26.5) at the 2 ppb, 72.9%(CV=18.2%) at the 4 ppb. (4) The preservation method for 6 estrogens was improved for the fast running time(21 min) and MSTFA was utilized for derivatizing 6 estrogens for improvement of recovery, for good resolution, for characteristic mass spectra unlike Jose's method and Tina's method. The utilization of zearalane as internal standard showed good quantification result for zeranol, zearalenone, as well as their isomers and metabolites, in beef.

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ZEARALENONE INDUCES MALE GERM CELL APOPTOSIS IN RATS

  • Kim, Il-Hwan;Kim, Yong-Bum;Son, Hwa-Young;Cho, Sung-Whan;Ha, Chang-Su;Kang, Boo-Hyon
    • Proceedings of the Korean Society of Toxicology Conference
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    • pp.96-96
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    • 2002
  • Zearalenone (ZEA), a nonsteroidal estrogenic mycotoxin, is known to cause toxicity of testis in male rats. To investigate whether apoptosis is involved in ZEA-induced testicular toxicity and to identify the stage and target germ cell type, 10-weeks-old Sprague-Dawley male rats were treated with a single intraperitoneal (i.p.) dose of ZEA (5mg/kg) and euthanized at 3, 6, 12, 24, and 48 h subsequently.(omitted)

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A Large Genomic Deletion in Gibberella zeae Causes a Defect in the Production of Two Polyketides but not in Sexual Development or Virulence

  • Lee Sun-Hee;Kim Hee-Kyoung;Hong Sae-Yeon;Lee Yin-Won;Yun Sung-Hwan
    • The Plant Pathology Journal
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    • v.22 no.3
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    • pp.215-221
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    • 2006
  • Gibberella zeae (anamorph: Fusarium graminearum) is an important pathogen of cereal crops. This fungus produces a broad range of secondary metabolites, including polyketides such as aurofusarin (a red pigment) and zearalenone (an estrogenic mycotoxin), which are important mycological characteristics of this species. A screen of G. zeae insertional mutants, generated using a restriction enzyme-mediated integration (REMI) procedure, led to the isolation of a mutant (Z43R606) that produced neither aurofusarin nor zearalenone yet showed normal female fertility and virulence on host plants. Outcrossing analysis confirmed that both the albino and zearalenone-deficient mutations are linked to the insertional vector in Z43R606. Molecular characterization of Z43R606 revealed a deletion of at least 220 kb of the genome at the vector insertion site, including the gene clusters required for the biosynthesis of aurofusarin and zearalenone, respectively. A re-creation of the insertional event of Z43R606 in the wild-type strain demonstrated that the 220-kb deletion is responsible for the phenotypic changes in Z43R606 and that a large region of genomic DNA can be efficiently deleted in G. zeae by double homologous recombination. The results showed that 52 putative genes located in the deleted genomic region are not essential for phenotypes other than the production of both aurofusarin and zearalenone. This is the first report of the molecular characterization of a large genomic deletion in G. zeae mediated by the REMI procedure.