• Title/Summary/Keyword: Aflatoxin %24M_1%24

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효소면역측정법에 의한 우유중의 Aflatoxin M$_{1}$ 분석

  • 손동화;임선희;이인원
    • Microbiology and Biotechnology Letters
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    • v.24 no.5
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    • pp.630-635
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    • 1996
  • For a survey of the occurrence of aflatoxin M$_{1}$ (AFM$_{1}$) in domestic cow's milk, we developed an enzyme-linked immunosorbent assay (ELISA) system, and quantitated the toxin in cow's milk. In order to produce specific antibodies AFM, conjugated to bovine serum albumin (AFM$_{1}$-BSA) and Freund's adjuvant were immunized subcutaneously to rabbits. By use of the antiserum showing the highest titer and AFB$_{1}$-HRP conjugate, we established a competitive direct ELISA (cdELISA) for AFM$_{1}$, whose detection limit was 0.003 ppb. The cross-reactivities of the antiserum against aflatoxin M$_{1}$ M$_{2}$, B$_{1}$, B$_{2}$, G$_{1}$, G$_{2}$, B$_{2a}$, and G$_{2a}$, were 100, 29.9, 25.0, 2.7, 13.0, 0.65, 0, and 0%, respectively. When the cdELISA was applied to the cow's milk spiked with AFM$_{1}$ and followed by cleanup with C$_{18}$ cartridge, the mean recovery of the assay was 104% (mean of CV, 6.4%) in the final concentration of 0.01-1 ppb (10-1, 000 ppt). When cow's milk samples gathered from markets and farms were assayed by the cdELISA, the mean concentration and SD of AFM$_{1}$ was 80.4 $\pm$ 55.0 ppt (n=64; range, 5.6-280 ppt).

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Occurrence and Estimation Using Monte-Carlo Simulation of Aflatoxin $M_1$in Domestic Cow’s Milk and Milk Products (국내산 우유 및 유제품에서의 Aflatoxin $M_1$오염수준 및 Monte-Carlo Simulation을 이용한 발생 추정)

  • 박경진;이미영;노우섭;천석조;심추창;김창남;신은하;손동화
    • Journal of Food Hygiene and Safety
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    • v.16 no.3
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    • pp.200-205
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    • 2001
  • In this study, occurrence of aflatoxin M$_1$(AEM$_1$) in domestic milk and milk products was determined. The level of AFM$_1$ in market milk (0.047 ppb) was lower than that in raw milk (0.083 pub) but this looks like that is due to dilution in collecting process rather than the effect of sterilization. In the case of nonfat dry milk, level of AFM$_1$appeared high by 0.24 ppb but it is thought to be not different from market milk actually because nonfat dry milk is diluted at intake. In the case of ice cream, finished products were contaminated with AFM$_1$of 0.020 ppd and also have the possibility of the contamination of AFB$_1$due to secondary raw material such as nuts and almond. On the basis of the results of this study and previous studies, Monte-Carlo simulation is conducted to estimate the contamination level of AFM$_1$in domestic market milk. To consider uncertainty and variability fitting procedure was passed through. And we used beta distribution to estimate the prevalence and triangular distribution to estimate the concentration level of AFM$_1$in milk. As a result, the 5%, 50% and 95% points of the distribution of the probability of AFM$_1$contamination level in milk is 0.0214, 0.0946 and 0.1888 ppb, respectively. Also we estimate that AFM$_1$in almost milk was low more than 0.5 ppb that is American acceptable level but 80.4% exceeded far 0.05 ppb that is European standard.

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The Screening of Aflatoxin Producing Fungi from Commercial Meju and Soy Bean Paste in Western Gyeongnam by Immunoassay (면역분석기법을 이용한 서부경남 시판 메주 및 된장에서의 Aflatoxin 생성균 검색)

  • 박정현;강성조;오상석;정덕화
    • Journal of Food Hygiene and Safety
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    • v.16 no.4
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    • pp.274-279
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    • 2001
  • Generally, non-aflatoxigenic fungi, such as Aspergillus oryzae, and Aspergillus are main microflora in Korean traditional fermented foods including Meju and soybean paste, but sometimes, Aspergillus flavus and Aspergillus parasiticus can be contaminated and accumulated aflatoxins during fermentation and storage. So the screening of aflatoxigenic strains in fermented traditional food is very important to improve the sanitary quality of those foods. In this work, we screened aflatoxin producing fungi from commercial Meju and soybean paste in Western Gyeongnam by immunoassay. Samples were randomly purchased from market of the commercial Meju(10 EA) and soybean paste(20 EA) in nine areas of Western Gyeongnam. Of the samples collected,24 strains and 22 strains of Aspergillus sp. were isolated from Meju and soybean paste, respectively. The isolated strains were cultured on SLS media at $25^{\circ}C$ for 15 days. The cultured broth were extracted with ethyl acetate and were analysed to determine aflatoxin B$_1$(AFB$_1$) by direct competitive ELISA(DC-ELISA). Six strains(25%) isolated from Meju, and 2 strains(9%) isolated from saybean paste, were confined as aflatoxin producing strains. The average range of aflatoxin productivity of isolates from Meju was 54.6 $\pm$ 38.7 ng/ml and that from soybean paste was 11.1 $\pm$ 8.6 ng/ml, respectively. Among them, isolated strain No. M-5-4 produced a high level of AFBl and showed 98.26 ng/ml of AFB$_1$. Every isolates were also re-confined their AFB$_1$productivity by thin layer chromatography(TLC). The TLC results also showed same trend as DC-ELISA results. As the above results, the screening of hazard mycotoxigenic fungi from traditional fermented foods should be necessary for the safety and the application of HACCP system in the food manufactory in Korea.

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Protective Effects of Milk Thistle (Silybum marianum) against Aflatoxin B1 in Broiler Chicks

  • Chand, N.;Muhammad, Din;Durrani, F.R.;Qureshi, M. Subhan;Ullah, Sahibzada S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.7
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    • pp.1011-1018
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    • 2011
  • Aflatoxin-contaminated feed cause mortality, suppression of the immune system, reduced growth rates and losses in feed efficiency. This research study was planned to investigate the immunomodulatory and growth promoting effect of milk thistle as feed additive against aflatoxin $B_1$ in broiler chicks at NWFP Agricultural University Peshawar, Pakistan. Two hundred and forty (240) day old broilers chicks were randomly assigned into four major groups AfF, aflatoxin free feed; Aflatoxin $B_1$ was present in the feed at the levels of 80-520 ${\mu}g/kg$ of the feed in the remaining three groups. Aflatoxin contaminated feed was provided for 5 weeks. Group AfB was supplemented with toxin binder "Mycoad" at 3 g/kg of feed and group AfT was supplemented with milk thistle at10 g/kg of feed. Each group was further sub divided into two sub-groups, vaccinated against ND (Newcastle disease), IB (Infectious bronchitis) and IBD (Infectious bursal diseases) according to recommended schedule of vaccination or non vaccinated. Each sub group carried three replicates with 10 chicks per replicate. Chicks were reared in pens in an open sided house. Supplementary heat was provided to all the chicks during brooding period. Mean body weight gain and dressing percentage were significantly (p<0.05) higher in group AfF, followed by AfT, AfB and Af. Weight gain and dressing percentage was the same in group AfB and AfT, while it was significantly lower in group Af. Feed intake, breast, thigh and leg weight were found significantly (p<0.05) higher in group AfF, followed by AfB, AfT and Af. Significantly lower (better) FCR value was recorded in group AfT. Water intake was significantly (p<0.05) higher in group AfT and AfF as compared to other groups. Mortality was significantly (p<0.05) higher in group Af. Mean bursa and thymus weights were found significantly (p<0.05) higher in group AfF, AfB and AfT followed by Af, while higher spleen weight was recorded in group AfT. Mean antibody titer against ND, IB and IBD was significantly (p<0.05) higher in group AfT, as compared to other groups. It is concluded that milk thistle at 10 g/kg of feed could effectively be utilized as immunostimulant and growth promotant in the presence of immunosuppressant aflatoxin $B_1$ in the feed.

In-field evaluation of clinoptilolite feeding efficacy on the reduction of milk aflatoxin M1 concentration in dairy cattle

  • Katsoulos, Panagiotis D.;Karatzia, Maria A.;Boscos, Constantinos;Wolf, Petra;Karatzias, Harilaos
    • Journal of Animal Science and Technology
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    • v.58 no.7
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    • pp.24.1-24.7
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    • 2016
  • Background: Clinoptilolite is a natural zeolite with high adsorption capacity for polar mycotoxins such as aflatoxins. The efficacy of clinoptilolite in ameliorating the toxic effects of aflatoxicosis has been proven in monogastric animals, but there is no such evidence for ruminants. The aim of this study was to evaluate, under field conditions, whether the dietary administration of clinoptilolite in dairy cows could reduce the concentration of aflatoxin M1 ($AFM_1$) in bulk-tank milk, in farms with higher than or close to $0.05{\mu}g/kg$ of milk (European maximum allowed residual level). An objective of the present study was also to investigate the effect of particle size of clinoptilolite on aflatoxin binding. Methods: Fifteen commercial Greek dairy herds with AFM1 concentrations in bulk tank milk ${\geq}0.05{\mu}g/kg$ were selected. Bulk tank milk AFM1 was determined prior to the onset and on day 7 of the experiment. Clinoptilolite was added in the total mixed rations of all farms at the rate of 200 g per animal per day, throughout this period. Two different particle sizes of clinoptilolite were used; less than 0.15 mm in 9 farms (LC group) and less than 0.8 mm in 6 farms (HC group). Results: Clinoptilolite administration significantly reduced $AFM_1$ concentrations in milk in all farms tested at an average rate of 56.2 % (SD: 15.11). The mean milk $AFM_1$ concentration recorded on Day 7 was significantly (P < 0.001) lower compared to that of Day 0 ($0.036{\pm}0.0061$ vs. $0.078{\pm}0.0074{\mu}g/kg$). In LC group farms the reduction of milk $AFM_1$ concentration was significantly higher than HC group farms ($0.046{\pm}0.0074$ vs. $0.036{\pm}0.0061{\mu}g/kg$, P = 0.002). As indicated by the Pearson correlation, there was a significant and strong linear correlation among the milk $AFM_1$ concentrations on Days 0 and 7 (R = 0.95, P < 0.001). Conclusions: Dietary administration of clinoptilolite, especially of smallest particle size, at the rate of 200 g per cow per day can effectively reduce milk $AFM_1$ concentration in dairy cattle and can be used as a preventive measure for the amelioration of the risks associated with the presence of aflatoxins in the milk of dairy cows.

INDUCTION OF CYTOCHROME P-450 ASSOCIATED MONOOXYGENASE ACTIVITIES BY PHENOBARBITAL AND 3-METHYLCHOLANTHRENE IN PRIMARY CULTURES OF ADULT RAT HEPATOCYTES

  • Park, Seong-Kyu;Ha, Jong-Ryul;Kim, H.M.;Yang, K.H.
    • Toxicological Research
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    • v.3 no.1
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    • pp.1-8
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    • 1987
  • In vitro induction of cytochrome 450 associated monooxygenase activities by phenobarbital (PB) and 3-methylcholanthrene (MC) was investigated in primary cultures of adult rat hepatocytes. PB and MC were added to the culture 24 hr after the initial plating of hepatocytes. A signiftcant increase of the activities of 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase were observed in MC and PB treated culture. MC caused about 500% induction of the initial oxidation rates of both enzymes in 48 hr. However the PB maintained both enzyme activities close to the level of freshly isolated hepatocytes. Biphenyl 4-hydroxylase and aminopyrine N-demethylase activities were also induced by MC and PB. But the level of induction was less than that occuring with 7-ethoxycoumarin 0-deethylase and aryl hydrocarbon hydroxylase. When aflatoxin $B_1$ was added to the hepatocyte cultures which have been treated with MC or PB, it caused a significant increase of the unscheduled DNA synthesis at higher dose of aflatoxin $B_1$ as compared to those of untreated control hepatocyte cultures. The results suggest that microsomal enzyme activities can be selectively controlled preferably in hepatocyte cultures by the in vitro induction method. This principle may be useful for studying the metabolism and other toxicological studies.

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Survey of Yogurt Powder Storage in Ambient Export Countries A Safety Evaluation Standard Compliance and Comparative Analysis

  • Kim, Na-Kyeong;Park, Jung-Min;Lee, Jung-Hoon;Kim, Ha-Jung;Oh, Sejong;Imm, Jee-Young;Lim, Kwang-Sei;Kim, Jin-Man
    • Food Science of Animal Resources
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    • v.35 no.1
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    • pp.143-148
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    • 2015
  • Yogurt powder is fermented milk processed in the form of dry yogurt, and has advantages such as stability, storability, convenience, and portability. China and Vietnam are important export target countries because of the increased demand for dairy products. Therefore, we surveyed dairy product standardization in order to establish an export strategy. Lactic acid bacteria counts are unregulated in Korea and Vietnam. In China, lactic acid bacteria counts are regulated at $1{\times}10^6$ colonyforming units (CFU)/mL and detected at $6.24{\pm}0.33\;Log\;CFU/mL$. All three countries have regulated standards for total bacterial counts. In China, total bacterial counts of milk powder are regulated to n=5, c=2, m=50,000, M=200,000 and detected at $6.02{\pm}0.12\;Log\;CFU/mL$, exceeding the acceptable level. Lactic acid bacterial counts appeared to exceed total bacterial counts. Coliform group counts, Staphylococcus aureus, Listeria monocytogenes, and Salmonella species were not detected. Acidity is not regulated in Korea and Vietnam. In China, acidity was regulated to over $70^{\circ}T$ and detected $352.38{\pm}10.24^{\circ}T$. pH is unregulated in all three countries. pH was compared to that of general fermented milk, which is 4.2, and that of the sample was $4.28{\pm}0.01$. Aflatoxin levels are not regulated in Korea and China. In Vietnam, aflatoxin level is regulated at 0.05 ppb. Therefore, all ingredients of the yogurt powder met the safety standards. This data obtained in this study can be used as the basic data in assessing the export quality of yogurt powder.

Antimutagenic Effects of Boiled Water Extract and Tannin from Persimmon Leaves (감잎 열수추출물 및 감잎 탄닌의 항돌연변이 효과)

  • 박건영;문숙희
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.24 no.6
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    • pp.880-886
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    • 1995
  • Antimutagenic effects of boiled water extract and tannin from persimmon leaves were studied by using Ames test, spore rec assay and SOS chromotest. Strong antimutagenic activities toward aflatoxin B1(AFB1), dimethyl-aminobiphenyl(DMAB), N-methyl-N'-nitro-N-nitrosoguanidine(MNNG) and 4-nitroquinoline-1-oxide(4-NQO) were observed when boiled water extract and tannin from the persimmon leaves were added in the Salmonella typhimurium TA 100. In spore rec assay using Bacillus subtilis H17($rec^{+}$) and M45($rec^{-}$), boiled water extract and tannin from the persimmon leaves considerably inhibited the mutagenesis induce by MNNG. In SOS chromotest using Escherichia coli PQ37, these samples also exhibited strong antimutagenic activity toward 4-NQO. The tannin was more effective than boiled water extract of persimmon leaves in the antimutagenicity tests.

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A Study on the Physiological Effects and Dyeing Properties of the Extract of Fermented Indigo(Part II) (발효쪽 추출물의 생리적 기능과 염색특성(제2보))

  • 한신영;최석철
    • Journal of the Korean Society of Clothing and Textiles
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    • v.24 no.8
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    • pp.1146-1154
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    • 2000
  • The purpose of this study was to investigate the antimicrobial activity, antimutagenic and anticancer effects and dyeing properties of the fermented indigo extract. The methanol extract of fermented indigo showed a strong inhibition effect on Trich. mentagrophytes and antimutagenic activities against aflatoxin B$_1$(AFB$_1$) in the Ames test using Salmonella typhimurium TA 100. We also found in vitro anticancer effects of the methanol extract of fermented indigo and it was evaluated by using Clone M-3 mouse melanoma cells and A431 human epidermoid carcinoma cells and exerted little cytotoxity against 3T3-L1 embryo fibroblast cell. In the relationship between the K/S values of cotton and silk fabrics dyed with fermented indigo and dyeing repeating times, the K/S values became higher as the repeating times were increased. The K/S values were high when the fabrics were dyed at low temperture. The K/S values of cotton fabrics were higher than those of silk fabrics. Changes of surface color of silk fabrics were higher than those of cotton fabrics after water fastness test, laundering, irradiation and treatment of acidic and alkaline perspiration.

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Desmutagenic Effect of Water Extract from Areca catechu L. on the Mutagenicity of N-Methyl-N-Nitro-N'-Nitrosoguanidinein in E. coli PQ37 (N-methyl-N-nitro-N'-nitrosoguanidine의 변이원성에 대한 빈랑 물 추출물의 돌연변이 억제효과)

  • Wu, Wei Jie;Ahn, Byung-Yong
    • Korean Journal of Medicinal Crop Science
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    • v.20 no.1
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    • pp.16-19
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    • 2012
  • The desmutagenic activity of the water extract of Areca catechu L. on the mutagenicity induced by aflatoxin $B_1$ ($AFB_1$), N-methyl-N-nitro-N'-nitrosoguani-dine (MNNG), mitomycin C (MMC) and 4-nitroquinoline 1-oxide (4-NQO) was studied by using the SOS Chromotest with Escherichia coli PQ37. The inhibition rates of water extract of Areca catechu L. at concentration of $100{\mu}g/assay$ were 41.0%, 47%, 46%, and 32% against $AFB_1$, MNNG, MMC and 4-NQO, respectively. The water extract of Areca catechu L. was separated into methanol soluble and methanol insoluble parts. The methanol insoluble part exhibited higher inhibition effect than the methanol soluble part against the mutagenic activities of MNNG. Step-wise fractionation of methanol insoluble part was done to obtain methanol, ethyl acetate and water fractions. Among these fractions, water fraction had the strongest inhibitory effect of 45.0% against mutagenicities of MNNG. The inhibition rates of aqueous fraction of methanol-insoluble from water extracted Areca catechu L. at concentrations of 1.61, 16.13, 161.29 and $322.58{\mu}g/mL$ were 12.0%, 24.0%, 47.5% and 62.0%, respectively. The water fraction showed the inhibitory effects with dose response against the mutagenic activity induced by MNNG.