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Sample records for aflatoxin b1 biomarker

  1. Aflatoxin exposure during the first 1000 days of life in rural South Asia assessed by aflatoxin B1-lysine albumin biomarkers

    Science.gov (United States)

    Groopman, John D.; Egner, Patricia A.; Schulze, Kerry J.; Wu, Lee S-F; Merrill, Rebecca; Mehra, Sucheta; Shamim, Abu A.; Ali, Hasmot; Shaikh, Saijuddin; Gernand, Alison; Khatry, Subarna K.; LeClerq, Steven C.; West, Keith P.; Christian, Parul

    2015-01-01

    Aflatoxin B1 is a potent carcinogen, occurring from mold growth that contaminates staple grains in hot, humid environments. In this investigation, aflatoxin B1-lysine albumin biomarkers were measured by mass spectrometry in rural South Asian women, during the first and third trimester of pregnancy, and their children at birth and at two years of age. These subjects participated in randomized community trials of antenatal micronutrient supplementation in Sarlahi District, southern Nepal and Gaibandha District in northwestern Bangladesh. Findings from the Nepal samples demonstrated exposure to aflatoxin, with 94% detectable samples ranging from 0.45 to 2939.30 pg aflatoxin B1-lysine/mg albumin during pregnancy. In the Bangladesh samples the range was 1.56 to 63.22 pg aflatoxin B1-lysine/mg albumin in the first trimester, 3.37 to 72.8 pg aflatoxin B1-lysine/mg albumin in the third trimester, 4.62 to 76.69 pg aflatoxin B1-lysine/mg albumin at birth and 3.88 to 81.44 pg aflatoxin B1-lysine/mg albumin at age two years. Aflatoxin B1-lysine adducts in cord blood samples demonstrated that the fetus had the capacity to convert aflatoxin into toxicologically active compounds and the detection in the same 2-year-old children illus trates exposure over the first 1000 days of life. PMID:25308602

  2. Fungal degradation of aflatoxin B1.

    Science.gov (United States)

    Shantha, T

    1999-01-01

    A number of fungal cultures were screened to select an organism suitable to be used in the detoxification of aflatoxin B1. They were co-cultured in Czapek-Dox-Casamino acid medium with aflatoxin B1 producing Aspergillus flavus. Several fungal cultures were found to prevent synthesis of aflatoxin B1 in liquid culture medium. Among these Phoma sp., Mucor sp., Trichoderma harzianum, Trichoderma sp. 639, Rhizopus sp. 663, Rhizopus sp. 710, Rhizopus sp. 668, Alternaria sp. and some strains belonging to the Sporotrichum group (ADA IV B14(a), ADA SF VI BF (9), strain 720) could inhibit aflatoxin synthesis by > or =90%. A few fungi, namely ADA IV B1, ADA F1, ADA F8, also belonging to the Sporotrichum group, were less efficient than the Phoma sp. The Cladosporium sp. and A. terreus sp. were by far the least efficient, registering aflatoxin biosynthesis are also capable of degrading the preformed toxin. Among these, Phoma sp. was the most efficient destroying about 99% of aflatoxin B1. The cell free extract of Phoma sp. destroyed nearly 50 microg aflatoxin B1 100 ml(-1) culture medium (90% of the added toxin), and this was more effective than its own culture filtrate over 5 days incubation at 28+/-2 degrees C. The degradation was gradual: 35% at 24 h, 58% at 48 h, 65% at 72 h, 85% at 96 h and 90% at 120 h. The possibility of a heat stable enzymatic activity in the cell free extract of Phoma is proposed. PMID:10945479

  3. Effects of Ginkgo biloba extract on expression of biomarkers during aflatoxin B1-induced hepatocarcinogenesis in Wistar rats

    Institute of Scientific and Technical Information of China (English)

    Yanrong Hao; Jianjia Su; Chao Ou; Ji Cao; Fang Yang; Xiaoxian Duan; Chun Yang; Yuan Li

    2012-01-01

    Objective: The aim of this study was to study the effect of Ginkgo biloba extract (EGb761) on metabolism of aflatoxin B1 (AFB1) in Wistar rats. Methods: Seventy one Wistar rats were assigned at random to groups A, B and C. Rats in groups A, B were injected with AFB1 (intraperitoneal, 100–200 μg/kg body weight, 1–3 times/week). Group C was normal control. Rats in group B were fed in food with EGb761, while rats in groups A, C were given normal food. Blood samples were collected and liver biopsies were performed on the 14th, 28th and 42nd week. All the rats were sacrificed on the 64th week. The incidence of hepatocarcinoma was investigated. The hepatic phase I drug-metabolizing enzyme Cytochrome-P450 (CYP450) and phase II metabolizing enzyme glutathione S-transferase (GST) were analyzed with spectrometry. Serum AFB1-lysine adduct levels were assessed with high performance liquid chromatography (HPLC). The expression of 8-hydroxydeoxy-guanosine (8-OHdG) was measured with immunohistochemistry. Results: The incidence of hepatocellular carcinoma (HCC) in group B was significantly lower than that in group A (26.92% vs 76.00%, P < 0.001). No HCC developed in group C. EGb761 showed no effects on the activities of CYP450 and GST in rat liver tissues. The level of AFB1-lysine adduct reached the peak (4356.01 pg/mg albumin) at the 14th week in group A. EGb761 significantly inhibited the formation of AFB1-lysine adduct in serum by 13.07% at the 14th week (P = 0.033), and 73.63% at the 42nd week (P = 0.002). The expression of 8-OHdG protein in rat liver tissues in group B was significantly lower than that in group A at the 28th, 42nd, and 64th week (P < 0.05). Conclu-sion: The main mechanism underlying the effect of EGb761 in blocking hepatocarcinogenesis induced by AFB1 may not be fully attributable to its influence on the activity of liver phase I and phase II metabolizing enzymes. EGb761 inhibits the produc-tion of AFB1-lysine adducts, decreases the expression of 8-OHd

  4. Subchronic mycotoxicoses in Wistar rats: assessment of the in vivo and in vitro genotoxicity induced by fumonisins and aflatoxin B(1), and oxidative stress biomarkers status.

    Science.gov (United States)

    Theumer, M G; Cánepa, M C; López, A G; Mary, V S; Dambolena, J S; Rubinstein, H R

    2010-01-31

    Some evidence suggests that fumonisin B(1) (FB(1)), a worldwide toxic contaminant of grains produced by Fusarium verticillioides, exhibits an oxidative stress mediated genotoxicity. We studied the DNA damage (by the alkaline comet and the micronucleus tests) and biomarkers of cellular oxidative stress (malondialdehyde, MDA; catalase, CAT; and superoxide dismutase, SOD) in spleen mononuclear cells of male Wistar rats subchronically (90 days) fed on a control experimental diet (CED) or poisoned with experimental diets contaminated with a culture material containing 100 ppm of FB(1) (FED), with 40 ppb of aflatoxin B(1) (a common toxic co-contaminant in cereals, AFB(1)ED), and with a mixture of both toxins (MED). The DNA damage was found in 13.7%, 81.7%, 98.0% and 99.3% (comet assay) and in 2.8%, 7.0%, 10.8% and 8.8% (micronucleus technique) in groups CED, FED, AFB(1)ED and MED, respectively. The MDA levels as well as the CAT and SOD activities were increased in all the poisoned animals. A similar behavior was observed in cells exposed in vitro to the toxins. These data support the hypothesis of an oxidative stress mediated genotoxicity induced by FB(1). Furthermore, the extent of DNA damage assessed by the comet assay suggests a possible protective effect of the fumonisins-AFB(1) mixtures in vitro against the genotoxicity induced individually by the toxins.

  5. Aflatoxin B1: Mechanism of mutagenesis

    Directory of Open Access Journals (Sweden)

    Regina M. Santella

    2007-02-01

    Full Text Available

    Aflatoxins are a group of toxic and carcinogenic fungal metabolites that frequently contaminate corn, peanuts and other products. Aflatoxin B1 (AFB1, the most potent of these, is metabolized by the cytochrome P450 system into a number of hydroxylated metabolites and glutathione conjugates in the process of conversion to more hydrophilic forms for urinary excretion. Unfortunately, one of these metabolites is the aflatoxin-8,9-epoxide that is produced in two forms, endo and exo. Glutathione S-transferases (GST are able to conjugate and detoxify this reactive intermediate. Species differences in susceptibility to the effects of AFB1 are partially related to differences in expression of specific GSTs that are able to conjugate the epoxide to glutathione. The exo epoxide is able to intercalate into DNA and this is followed by reaction of the C8 position of the epoxide with the N7 position of guanine.

    NMR studies of oligonucleotide duplexes containing the adduct have demonstrated that the adduct exists with the aromatic portion intercalated on the 5' face of the guanine residue with Watson-Crick base pairing maintained.

    However, this covalent adduct is chemically unstable due to the charge on the ribose ring. As a result, the guanine can be released from the DNA leaving an apurinic site. This released guanine adduct can be detected in the urine and serves as a biomarker of exposure to AFB1. Alternatively, the ribose ring opens forming a stable formamidopyrimidine (FAPY adduct. This adduct somewhat stabilizes the DNA duplex. Time course studies in animals have demonstrated that the N7 adduct is rapidly removed, probably because it causes more distortion in the helix, while the FAPY adduct is more

  6. Compound list: aflatoxin B1 [Open TG-GATEs

    Lifescience Database Archive (English)

    Full Text Available aflatoxin B1 AFB1 00165 ftp://ftp.biosciencedbc.jp/archive/open-tggates/LATEST/Human/in_vitro/aflat...oxin_B1.Human.in_vitro.Liver.zip ftp://ftp.biosciencedbc.jp/archive/open-tggates/LATEST/Rat/in_vivo/Liver/Single/aflatoxin_B1.Rat.in_vivo.Liver.Single.zip ...

  7. Surface Binding of Aflatoxin B1 by Lactic Acid Bacteria

    OpenAIRE

    Haskard, Carolyn A.; El-Nezami, Hani S.; Kankaanpää, Pasi E.; Salminen, Seppo; Ahokas, Jorma T.

    2001-01-01

    Specific lactic acid bacterial strains remove toxins from liquid media by physical binding. The stability of the aflatoxin B1 complexes formed with 12 bacterial strains in both viable and nonviable (heat- or acid-treated) forms was assessed by repetitive aqueous extraction. By the fifth extraction, up to 71% of the total aflatoxin B1 remained bound. Nonviable bacteria retained the highest amount of aflatoxin B1. Lactobacillus rhamnosus strain GG (ATCC 53103) and L. rhamnosus strain LC-705 (DS...

  8. Occurrence of aflatoxin B1 in natural products

    OpenAIRE

    Guilherme Prado; Altoé, Aline F.; Tatiana C. B. Gomes; Leal, Alexandre S.; Morais, Vanessa A. D.; Marize S. Oliveira; Marli B. Ferreira; Gomes, Mateus B.; Fabiano N. Paschoal; Rafael von S. Souza; Daniela A. Silva; Cruz Madeira, Jovita E. G.

    2012-01-01

    The media claims for the consumption of natural resource-based food have gradually increased in both developing and developed countries. The interest in the safety of these products is partially due to the possible presence of toxigenic fungi acting as mycotoxin producers, such as aflatoxins produced during the secondary metabolism of Aspergillus flavus, A. parasiticus and A. nomius. Aflatoxins, mainly aflatoxin B1, are directly associated with liver cancer in human beings. This paper is aime...

  9. Occurrence of B1 Aflatoxin in diet and M1 Aflatoxin in bovine milk

    OpenAIRE

    Adriana Frizzarin; Thiago Pereira Motta; Thamires Martins; Livia Castelani; Heloisa Solda de Azevedo; Cláudia Rodrigues Pozzi

    2012-01-01

    Ensuring food quality is one of the principles of food safety. Food for dairy cattle may be contaminated by fungi of the genus Aspergillus, which produce aflatoxins. The B1 aflatoxin, when ingested by animals, is biotransformed in liver in several other toxic metabolites, including M1 aflatoxin which is excreted in milk. M1 aflatoxin has a carcinogenic effect, which the presence in milk poses a serious risk to public health because milk and dairy products are consumed mainly by children, preg...

  10. Occurrence of aflatoxin B1 in natural products.

    Science.gov (United States)

    Prado, Guilherme; Altoé, Aline F; Gomes, Tatiana C B; Leal, Alexandre S; Morais, Vanessa A D; Oliveira, Marize S; Ferreira, Marli B; Gomes, Mateus B; Paschoal, Fabiano N; von S Souza, Rafael; Silva, Daniela A; Cruz Madeira, Jovita E G

    2012-10-01

    The media claims for the consumption of natural resource-based food have gradually increased in both developing and developed countries. The interest in the safety of these products is partially due to the possible presence of toxigenic fungi acting as mycotoxin producers, such as aflatoxins produced during the secondary metabolism of Aspergillus flavus, A. parasiticus and A. nomius. Aflatoxins, mainly aflatoxin B1, are directly associated with liver cancer in human beings. This paper is aimed at evaluating the presence of aflatoxin B1 in a few vegetable drugs, dried plant extracts and industrialized products traded in 2010 in the city of Belo Horizonte, State of Minas Gerais, Brazil. The method used for the quantification of aflatoxin B1 was based on extraction through acetone:water (85:15), immunoaffinity column purification followed by separation and detection in high efficiency liquid chromatography. Under the conditions of analysis, the Limits of Detection and Quantification were 0.6 µg kg(-1) and 1.0 µg kg(-1) respectively. The complete sets of analyses were carried out in duplicate. Aflatoxin B1 was noticed in a single sample (aflatoxin B1 contamination in the products under analysis. However, it is required to establish a broad monitoring program in order to obtain additional data and check up on the actual extension of contamination. PMID:24031973

  11. Occurrence of aflatoxin B1 in natural products

    Directory of Open Access Journals (Sweden)

    Guilherme Prado

    2012-12-01

    Full Text Available The media claims for the consumption of natural resource-based food have gradually increased in both developing and developed countries. The interest in the safety of these products is partially due to the possible presence of toxigenic fungi acting as mycotoxin producers, such as aflatoxins produced during the secondary metabolism of Aspergillus flavus, A. parasiticus and A. nomius. Aflatoxins, mainly aflatoxin B1, are directly associated with liver cancer in human beings. This paper is aimed at evaluating the presence of aflatoxin B1 in a few vegetable drugs, dried plant extracts and industrialized products traded in 2010 in the city of Belo Horizonte, State of Minas Gerais, Brazil. The method used for the quantification of aflatoxin B1 was based on extraction through acetone:water (85:15, immunoaffinity column purification followed by separation and detection in high efficiency liquid chromatography. Under the conditions of analysis, the Limits of Detection and Quantification were 0.6 µg kg-1 and 1.0 µg kg-1respectively. The complete sets of analyses were carried out in duplicate. Aflatoxin B1 was noticed in a single sample (< 1.0 µg kg-1. The results revealed low aflatoxin B1contamination in the products under analysis. However, it is required to establish a broad monitoring program in order to obtain additional data and check up on the actual extension of contamination.

  12. Supercritical Fluid Extraction of Aflatoxin B 1 from Soil

    Science.gov (United States)

    This research describes the development of a Supercritical Fluid Extraction (SFE) method to recover aflatoxin B1 from fortified soil. The effects of temperature, pressure, modifier (identity and percentage), and extraction type were assessed. Using the optimized SFE conditions, ...

  13. Aspergillus flavus and aflatoxin B1 in flour production.

    Science.gov (United States)

    Halt, M

    1994-10-01

    This paper discusses the results of investigations of contamination with aflatoxin-producing fungi and aflatoxin B1 affecting 545 samples of wheat grains, 475 samples of intermediate products of wheat grain being milled to flour (like middlings) and 238 samples of flour. A significant contamination with moulds was detected in analyzed samples. Although Aspergillus (34.87%) and Penicillium (32.37%) dominated, other types were also present, e.g., Cladosporium, Fusarium, Mucor, Alternaria, Rhizopus, Absidia and Trichoderma (listed in order of frequency). The presence of Aspergillus flavus, the known aflatoxin producer, was detected in 9.94% of analyzed samples. Isolates of A. Flavus were capable of producing aflatoxin B1 under favourable conditions. Aflatoxin B1 was found in 76.8% of samples contaminated with A. flavus. The highest contamination with aflatoxin B1 was detected in wheat grain samples (mean value of 16.3 micrograms/kg) and in intermediate products of wheat grain being milled to flour (mean value of 11.13 micrograms/kg). Contamination was lower in flour samples (mean value of 4.13 micrograms/kg). With regard to proposed standards given by the FAO and WHO, under which the content of aflatoxin should not exceed 30 micrograms/kg in food products, only two of 96 samples did not meet these criteria. PMID:7859854

  14. Base substitution mutations induced by metabolically activated aflatoxin B1.

    Science.gov (United States)

    Foster, P L; Eisenstadt, E; Miller, J H

    1983-05-01

    We have determined the base substitutions generated by metabolically activated aflatoxin B1 in the lacI gene of a uvrB- strain of Escherichia coli. By monitoring over 70 different nonsense mutation sites, we show that activated aflatoxin B1 specifically induced GxC leads to TxA transversions. One possible pathway leading to this base change involves depurination at guanine residues. We consider this mechanism of mutagenesis in the light of our other findings that the carcinogens benzo[a]pyrene diol epoxide and N-acetoxyacetylaminofluorene also specifically induce GxC leads to TxA transversions. PMID:6405385

  15. Base substitution mutations induced by metabolically activated aflatoxin B1.

    OpenAIRE

    Foster, P. L.; Eisenstadt, E; Miller, J H

    1983-01-01

    We have determined the base substitutions generated by metabolically activated aflatoxin B1 in the lacI gene of a uvrB- strain of Escherichia coli. By monitoring over 70 different nonsense mutation sites, we show that activated aflatoxin B1 specifically induced GxC leads to TxA transversions. One possible pathway leading to this base change involves depurination at guanine residues. We consider this mechanism of mutagenesis in the light of our other findings that the carcinogens benzo[a]pyren...

  16. Dual effects of phloretin on aflatoxin B1 metabolism: activation and detoxification of aflatoxin B1.

    Science.gov (United States)

    Gao, Shang Shang; Chen, Xiao Yan; Zhu, Ri Zhe; Choi, Byung-Min; Kim, Sun Jun; Kim, Bok-Ryang

    2012-01-01

    Typically, chemopreventive agents involve either induction of phase II detoxifying enzymes and/or inhibition of cytochrome P450 enzymes (CYPs) that are required for the activation of procarcinogens. In this study, we investigated the protective effects of phloretin against aflatoxin B1 (AFB1) activation to the ultimate carcinogenic intermediate, AFB(1)-8, 9-epoxide (AFBO), and its subsequent detoxification. Phloretin markedly inhibited formation of the epoxide with human liver microsomes in a dose-dependent manner. Phloretin also inhibited the activities of nifedipine oxidation and ethoxyresorufin O-deethylase (EROD) in human liver microsomes. These data show that phloretin strongly inhibits CYP1A2 and CYP3A4 activities, which are involved in the activation of AFB1. Phloretin increased glutathione S-transferase (GST) activity of alpha mouse liver 12 (AML 12) cells in a dose-dependent manner. GST activity toward AFBO in cell lysates treated with 20 μM phloretin was 23-fold that of untreated control cell lysates. The expression of GSTA3, GSTA4, GSTM1, GSTP1 and GSTT1 was induced by phloretin in a dose-dependent manner in AML 12 cells. GSTP1, GSTM1, and GSTT1 were able to significantly increase the conjugation of AFBO with glutathione. Concurrently, induction of the GST isozyme genes was partially associated with the Nrf2/ARE pathway. Taken together, the results demonstrate that phloretin has a strong chemopreventive effect against AFB1 through its inhibitory effect on CYP1A2, CYP3A4, and its inductive effect on GST activity. PMID:22253071

  17. Occurrence of B1 Aflatoxin in diet and M1 Aflatoxin in bovine milk

    Directory of Open Access Journals (Sweden)

    Adriana Frizzarin

    2012-12-01

    Full Text Available Ensuring food quality is one of the principles of food safety. Food for dairy cattle may be contaminated by fungi of the genus Aspergillus, which produce aflatoxins. The B1 aflatoxin, when ingested by animals, is biotransformed in liver in several other toxic metabolites, including M1 aflatoxin which is excreted in milk. M1 aflatoxin has a carcinogenic effect, which the presence in milk poses a serious risk to public health because milk and dairy products are consumed mainly by children, pregnant women and elderly. The objective of this study was to detect the presence of B1 aflatoxin in feed supplied to dairy cows and the presence of M1 aflatoxin in milk. Samples were collected from complete diet (corn silage and concentrate from a batch of 15 lactating cows from a dairy farm in the Campinas region. Two samples of diets were collected directly into the troughs in intervals of 24 hours at every 15 days, totalizing a period of 45 days. Milk samples of those cows were collected 24 hours after diet collection, directly from sample valves in the glass jars.. B1 and M1 aflatoxins were detected by the technique of High Performance Liquid Chromatography after extraction and purification on immunoaffinity columns. From the 40 samples of diets evaluated, 40% were contaminated with B1 aflatoxin, and the levels found ranged from 1.93 to 43.78μg/Kg. One sample showed result higher than the maximum recommended for grain and animal feed in Brazil (20μg/Kg. From the 75 milk samples analyzed, the presence of M1 aflatoxin was detected in 13.3% with levels ranging from 0.03 to 0.16μg/L, not exceeding the maximum permitted for marketing in the country of 0.5μg/L, however 80% of contaminated samples had values above the maximum permissible levels of 0.05μg/L, value found among countries with abundant milk production... The presence of aflatoxins highlights the importance of monitoring the production, the storage and the importance of handling food and

  18. AFLATOXIN B1 RESIDUES IN EGGS AND FLESH OF LAYING HENS FED AFLATOXIN B1 CONTAMINATED DIET

    Directory of Open Access Journals (Sweden)

    Saqer Mohammad Herzallah

    2013-01-01

    Full Text Available Aflatoxin B1(AFB1 and total Aflatoxins (AFT contaminated feed effect on aflatoxins residue level in eggs, muscles (breast, leg, organs (liver, kidney, gizzard and excreted aflatoxins in chicken litter of layer hens were monitored. Laying hens were on four levels of aflatoxins for 6 weeks and monitored weekly for the change in both AFB1 and AFT levels. Pronouncedly, the AFB1 and AFT were detected in eggs, muscles (legs, breast, organs (liver, kidney and gizzard and litter in noticeable amounts. Total Aflatoxin (AFT level was lowest in chicken breast (0.63 ppb and highest in liver (2.12 ppb and gizzard (1.22 ppb of chicken fed diet with 965.12 ppb. Whereas, AFB1 residue was 0.66 ppb in eggs, 1.59 ppb in liver tissues of hens given feed contaminated with 894.12 ppb. Residue level of AFB1 was high in liver and kidney of all treatments. The chicken breast tissues were lowest in AFB1 and AFT of values 0.72 and 0.63, respectively. Eggs production was significantly (p<0.05 affected with AFB1 contaminated feed and egg production was decreased by more than 30%.

  19. Rapid Immunoenzyme Assay of Aflatoxin B1 Using Magnetic Nanoparticles

    OpenAIRE

    Urusov, Alexandr E.; Alina V. Petrakova; Vozniak, Maxim V.; Zherdev, Anatoly V.; Boris B. Dzantiev

    2014-01-01

    The main limitations of microplate-based enzyme immunoassays are the prolonged incubations necessary to facilitate heterogeneous interactions, the complex matrix and poorly soluble antigens, and the significant sample dilutions often required because of the presence of organic extractants. This study presents the use of antibody immobilization on the surface of magnetic particles to overcome these limitations in the detection of the mycotoxin, aflatoxin B1. Features of the proposed system are...

  20. Aflatoxin B1 in poultry: toxicology, metabolism and prevention.

    Science.gov (United States)

    Rawal, Sumit; Kim, Ji Eun; Coulombe, Roger

    2010-12-01

    Aflatoxins (AF) are ubiquitous in corn-based animal feed and causes hepatotoxic and hepatocarcinogenic effects. The most important AF in terms of toxic potency and occurrence is aflatoxin B1 (AFB1). Poultry, especially turkeys, are extremely sensitive to the toxic and carcinogenic action of AFB1, resulting in millions of dollars in annual losses to producers due to reduced growth rate, increased susceptibility to disease, reduced egg production and other adverse effects. The extreme sensitivity of turkeys and other poultry to AFB1 is associated with efficient hepatic cytochrome P450-mediated bioactivation and deficient detoxification by glutathione S-transferases (GST). Discerning the biochemical and molecular mechanisms of this extreme sensitivity of poultry to AFB1, will contribute in the development of novel strategies to increase aflatoxin resistance. Since AFB1 is an unavoidable contaminant of corn-based poultry feed, chemoprevention strategies aimed at reducing AFB1 toxicity in poultry and in other animals have been the subject of numerous studies. This brief review summarizes many of the key recent findings regarding the action of aflatoxins in poultry.

  1. Determination of aflatoxin B1 in medical herbs: interlaboratory study.

    Science.gov (United States)

    Arranz, Isabel; Sizoo, Eric; van Egmond, Hans; Kroeger, Katy; Legarda, Teresa M; Burdaspal, Pedro; Reif, Klaus; Stroka, Joerg

    2006-01-01

    A method was developed for the determination of aflatoxin B1 in medical herbs (senna pods, botanical name Cassia angustifolia; devil's claw, botanical name Harpagophytum procumbens; and ginger roots, botanical name Zingiber officinale). The method, which was tested in a mini-collaborative study by 4 laboratories, is based on an immunoaffinity cleanup followed by reversed-phase high-performance liquid chromatography separation and fluorescence detection after post-column derivatization. It allows the quantitation of aflatoxin B1 at levels lower than 2 ng/g. A second extractant (acetone-water) was tested and compared to the proposed methanol-water extractant. Several post-column derivatization options (electrochemically generated bromine, photochemical reaction, and chemical bromination) as well as different integration modes (height versus area) were also investigated. No differences were found depending on the choice of derivatization system or the signal integration mode used. The method was tested for 3 different matrixes: senna pods, ginger root, and devil's claw. Performance characteristics were established from the results of the study and resulted in HorRat values ranging from 0.12 to 0.75 with mean recoveries from 78 to 91% for the extraction with methanol-water and HorRat values ranging from 0.10-1.03 with mean recoveries from 98 to 103% for the extraction with acetone-water. As a result, the method, with all tested variations, was found to be fit-for-purpose for the determination of aflatoxin B1 in medical herbs at levels of 1 microg/kg and above. PMID:16792057

  2. Rapid Immunoenzyme Assay of Aflatoxin B1 Using Magnetic Nanoparticles

    Directory of Open Access Journals (Sweden)

    Alexandr E. Urusov

    2014-11-01

    Full Text Available The main limitations of microplate-based enzyme immunoassays are the prolonged incubations necessary to facilitate heterogeneous interactions, the complex matrix and poorly soluble antigens, and the significant sample dilutions often required because of the presence of organic extractants. This study presents the use of antibody immobilization on the surface of magnetic particles to overcome these limitations in the detection of the mycotoxin, aflatoxin B1. Features of the proposed system are a high degree of nanoparticle dispersion and methodologically simple immobilization of the antibodies by adsorption. Reactions between the immobilized antibodies with native and labeled antigens are conducted in solution, thereby reducing the interaction period to 5 min without impairing the analytical outcome. Adsorption of immunoglobulins on the surface of magnetic nanoparticles increases their stability in aqueous-organic media, thus minimizing the degree of sample dilution required. Testing barley and maize extracts demonstrated a limit of aflatoxin B1 detection equal to 20 pg/mL and total assay duration of 20 min. Using this method, only the 3-fold dilution of the initial methanol/water (60/40 extraction mixture in the microplate wells is necessary. The proposed pseudo-homogeneous approach could be applied toward immunodetection of a wide range of compounds.

  3. Rapid immunoenzyme assay of aflatoxin B1 using magnetic nanoparticles.

    Science.gov (United States)

    Urusov, Alexandr E; Petrakova, Alina V; Vozniak, Maxim V; Zherdev, Anatoly V; Dzantiev, Boris B

    2014-01-01

    The main limitations of microplate-based enzyme immunoassays are the prolonged incubations necessary to facilitate heterogeneous interactions, the complex matrix and poorly soluble antigens, and the significant sample dilutions often required because of the presence of organic extractants. This study presents the use of antibody immobilization on the surface of magnetic particles to overcome these limitations in the detection of the mycotoxin, aflatoxin B1. Features of the proposed system are a high degree of nanoparticle dispersion and methodologically simple immobilization of the antibodies by adsorption. Reactions between the immobilized antibodies with native and labeled antigens are conducted in solution, thereby reducing the interaction period to 5 min without impairing the analytical outcome. Adsorption of immunoglobulins on the surface of magnetic nanoparticles increases their stability in aqueous-organic media, thus minimizing the degree of sample dilution required. Testing barley and maize extracts demonstrated a limit of aflatoxin B1 detection equal to 20 pg/mL and total assay duration of 20 min. Using this method, only the 3-fold dilution of the initial methanol/water (60/40) extraction mixture in the microplate wells is necessary. The proposed pseudo-homogeneous approach could be applied toward immunodetection of a wide range of compounds. PMID:25412219

  4. Analysis of aflatoxins B1 and G1 in maize by quechers

    OpenAIRE

    Bursić Vojislava P.; Vuković Gorica Lj.; Jajić Igor M.; Lazić Sanja D.; Kara Magdalena H.; Čolović Radmilo R.; Vukmirović Đuro M.

    2013-01-01

    A reliable and easy method has been developed for the determination of aflatoxins B1 and G2 in maize samples. High performance liquid chromatography coupled with FLD (HPLC-FLD) with photochemical derivatization was used. Mycotoxins were extracted from maize using a QuEChERS-based extraction procedure. The optimized analytical conditions were evaluated in terms of recoveries, reproducibility, LOD, LOQ and linearity for aflatoxin B1 and aflatoxin G1 in maize. Extraction, chromatographic a...

  5. BIOCONTROL OF ASPERGILLUS FLAVUS AND AFLATOXIN B1 PRODUCTION IN CORN

    OpenAIRE

    A. Khanafari, H. Soudi, M. Miraboulfathi

    2007-01-01

    The potent mycotoxin aflatoxin B1 is a secondary metabolite of Aspergillus fungi that grow, on a variety of food and feed commodities at any stage during growth, harvest, storage and transportation. The occurrence of aflatoxin contamination is global, with severe problems especially prevalent in developing countries. In present study, corn samples were contaminated with aflatoxin B1 in the concentration of 240 µg/kg. Four trials were inoculated by Lactobacillus plantarum (PTCC 1058). Three co...

  6. Emericella astellata, a new producer of aflatoxin B-1, B-2 and sterigmatocystin

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Samson, R.A.; Smedsgaard, Jørn

    2004-01-01

    To report on aflatoxin B-1 and B-2 production from a species of Emericella. Methods and Results: Aflatoxins and sterigmatocystin were determined by high-pressure liquid chromatography (HPLC) with diode array detection and confirmed by HPLC with mass spectrometry detection. Among 30 known species...... of Emericella only one species produced aflatoxin. Strains originating from the same geographical source material had different patterns of aflatoxin and sterigmatocystin production on different media, indicating that epigenetic factors may be involved in the regulation of aflatoxin production. However, two...... cultures from the same original genet were very similar. Conclusions: Emericella astellata can produce small amounts of sterigmatocystin and aflatoxin B-1 and B-2. Significance and Impact of the Study: Emericella has been used extensively in genetic studies and therefore the isolates producing aflatoxin...

  7. Determination of Aflatoxin B1 in Smokeless Tobacco Products by Use of UHPLC-MS/MS.

    Science.gov (United States)

    Zitomer, Nicholas; Rybak, Michael E; Li, Zhong; Walters, Matthew J; Holman, Matthew R

    2015-10-21

    This work developed a UHPLC-MS/MS method for the detection and quantitation of aflatoxins in smokeless tobacco products, which was then used to determine aflatoxin B1 concentrations in 32 smokeless tobacco products commercially available in the United States. Smokeless tobacco products were dried, milled, and amended with (13)C17-labeled internal standards, extracted in water/methanol solution in the presence of a surfactant, isolated through use of immunoaffinity column chromatography, and reconstituted in mobile phase prior to UHPLC-MS/MS analysis. The method was capable of baseline separation of aflatoxins B1, B2, G1, and G2 in a 2.5 min run by use of a fused core C18 column and a water/methanol gradient. MS/MS transition (m/z) 313.3 → 241.2 was used for aflatoxin B1 quantitation, with 313.3 → 285.1 used for confirmation. The limit of detection (LOD) for aflatoxin B1 was 0.007 parts per billion (ppb). Method imprecision for aflatoxin B1 (expressed as coefficient of variation) ranged from 5.5 to 9.4%. Spike recoveries were 105-111%. Aflatoxin B1 concentrations in the smokeless tobacco products analyzed ranged from snuffs and chews, whereas all moist snuff products tested were below LOD. The amounts of aflatoxin B1 detected were low relative to the 20 ppb regulatory limit established by the U.S. Food and Drug Administration for foods and feeds.

  8. Aflatoxin B1 Degradation by a Pseudomonas Strain

    Science.gov (United States)

    Sangare, Lancine; Zhao, Yueju; Folly, Yawa Minnie Elodie; Chang, Jinghua; Li, Jinhan; Selvaraj, Jonathan Nimal; Xing, Fuguo; Zhou, Lu; Wang, Yan; Liu, Yang

    2014-01-01

    Aflatoxin B1 (AFB1), one of the most potent naturally occurring mutagens and carcinogens, causes significant threats to the food industry and animal production. In this study, 25 bacteria isolates were collected from grain kernels and soils displaying AFB1 reduction activity. Based on its degradation effectiveness, isolate N17-1 was selected for further characterization and identified as Pseudomonas aeruginosa. P. aeruginosa N17-1 could degrade AFB1, AFB2 and AFM1 by 82.8%, 46.8% and 31.9% after incubation in Nutrient Broth (NB) medium at 37 °C for 72 h, respectively. The culture supernatant of isolate N17-1 degraded AFB1 effectively, whereas the viable cells and intra cell extracts were far less effective. Factors influencing AFB1 degradation by the culture supernatant were investigated. Maximum degradation was observed at 55 °C. Ions Mn2+ and Cu2+ were activators for AFB1 degradation, however, ions Mg2+, Li+, Zn2+, Se2+, Fe3+ were strong inhibitors. Treatments with proteinase K and proteinase K plus SDS significantly reduced the degradation activity of the culture supernatant. No degradation products were observed based on preliminary LC-QTOF/MS analysis, indicating AFB1 was metabolized to degradation products with chemical properties different from that of AFB1. The results indicated that the degradation of AFB1 by P. aeruginosa N17-1 was enzymatic and could have a great potential in industrial applications. This is the first report indicating that the isolate of P. aeruginosa possesses the ability to degrade aflatoxin. PMID:25341538

  9. Metabolism of aflatoxin B1 and identification of the major aflatoxin B1-DNA adducts formed in cultured human bronchus and colon

    DEFF Research Database (Denmark)

    1979-01-01

    Aflatoxin B1 and benzo(a)pyrene were activated by both cultured human bronchus and human colon as measured by binding to cellular DNA and protein. The binding of aflatoxin B1 to DNA was dose dependent, and the level of binding was higher in cultured human bronchus than it was in the colon. When...... compared to aflatoxin B1, the binding level of benzo(a)pyrene to both bronchial and colonic DNA was generally higher. The major adducts formed in both tissues by the interaction of aflatoxin B1 and DNA were chromatographically identical to 2,3-dihydro-2-(N7-guanyl)-3-hydroxyaflatoxin B1 (Structure 1...... in these two peaks, and the ratio of radioactivity between the peaks was nearly 1. In colonic DNA, the ratio between Structures 1 and 11 was approximately 2. These observations add aflatoxin B1 to the list of chemical procarcinogens metabolized by cultured human tissues and in which the carcinogen-DNA adducts...

  10. Aflatoxin B1 transfer and metabolism in human placenta.

    Science.gov (United States)

    Partanen, Heidi A; El-Nezami, Hani S; Leppänen, Jukka M; Myllynen, Päivi K; Woodhouse, Heather J; Vähäkangas, Kirsi H

    2010-01-01

    Aflatoxin B1 (AFB1), a common dietary contaminant, is a major risk factor of hepatocellular carcinoma (HCC). Early onset of HCC in some countries in Africa and South-East Asia indicates the importance of early life exposure. Placenta is the primary route for various compounds, both nutrients and toxins, from the mother to the fetal circulation. Furthermore, placenta contains enzymes for xenobiotic metabolism. AFB1, AFB1-metabolites, and AFB1-albumin adducts have been detected in cord blood of babies after maternal exposure during pregnancy. However, the role that the placenta plays in the transfer and metabolism of AFB1 is not clear. In this study, placental transfer and metabolism of AFB1 were investigated in human placental perfusions and in in vitro studies. Eight human placentas were perfused with 0.5 or 5microM AFB1 for 2-4 h. In vitro incubations with placental microsomal and cytosolic proteins from eight additional placentas were also conducted. Our results from placental perfusions provide the first direct evidence of the actual transfer of AFB1 and its metabolism to aflatoxicol (AFL) by human placenta. In vitro incubations with placental cytosolic fraction confirmed the capacity of human placenta to form AFL. AFL was the only metabolite detected in both perfusions and in vitro incubations. Since AFL is less mutagenic, but putatively as carcinogenic as AFB1, the formation of AFL may not protect the fetus from the toxicity of AFB1.

  11. Genotoxicity of aflatoxin B1 and its ammonium derivatives.

    Science.gov (United States)

    Márquez Márquez, R; Tejada de Hernandez, I; Madrigal Bujaidar, E

    1995-01-01

    Aflatoxin (AF) B1 is a main contaminant in diverse agricultural products. In an attempt to reduce this problem and the hazards to human health, an AFB1 inactivating system with ammonia has been developed. In this work we evaluated the efficiency of the system in mice using micronucleus (MN) and sister chromatid exchange (SCE) analysis. Four groups of animals were fed for 8 weeks with a special diet mainly composed of maize: (1) uncontaminated; (2) uncontaminated/inactivated; (3) contaminated/inactivated; and (4) contaminated. We evaluated MN at weekly intervals in peripheral blood, and in weeks 4 and 8 SCE frequencies were quantified in bone marrow cells. The results shows that animals fed with AFB1 contaminated/inactivated maize had a 45% lower level of induced cytogenetic damage than those animals fed with AFB1 contaminated but not inactivated maize. A residual amount of AFB1 after the inactivating treatment and reconversion back to AFB1 in the organism may account for the remaining increased levels of SCE and MN.

  12. Aflatoxin B1 metabolism by 3-methylcholanthrene-induced hamster hepatic cytochrome P-450s.

    Science.gov (United States)

    Lai, T S; Chiang, J Y

    1990-01-01

    We have studied the activation of aflatoxin B1 by hamster liver microsomes and purified hamster cytochrome P-450 isozymes using a umu mutagen test. The hamster liver microsomes or S-9 fractions were much more active than rat liver microsomes or S-9 fractions in the activation of umu gene expression by aflatoxin B1 metabolites. 3-Methyl-cholanthrene treatment increased aflatoxin B1 activation by hamster liver microsomes. Two major 3-methylcholanthrene-inducible cytochrome P-450 isozymes, P-450 MC1 (IIA) and P-450 MC4 (IA2), were purified from 3-methylcholanthrene-treated hamster liver microsomes, and the metabolism of aflatoxin B1 by these two cytochromes was studied. In the reconstituted enzyme system, both P-450 MC1 and P-450 MC4 were highly active in the activation of aflatoxin B1, and antibodies against these P-450s specifically inhibited these activities. Antibody against P-450 MC1 inhibited the activation of aflatoxin B1 by 20% in the presence of 3-methyl-cholanthrene-treated hamster liver microsomes. In contrast, antibody against P-450 MC4 stimulated the activity by 175%. These results indicated that hamster P-450 MC1 might convert aflatoxin B1 to more toxic metabolite(s), whereas P-450 MC4 might convert aflatoxin B1 to less toxic metabolite(s), than aflatoxin B1 in liver microsomes. The metabolite(s) produced by both hamster cytochrome P-450 MC1 and MC4 were genotoxic in the umu mutagen test. PMID:2126562

  13. Emericella venezuelensis, a new species with stellate ascospores producing sterigmatocystin and aflatoxin B-1

    DEFF Research Database (Denmark)

    Frisvad, Jens Christian; Samson, R.A.

    2004-01-01

    media. The three species also differ in their profiles of extrolites (secondary metabolites). Emericella venezuelensis produces aflatoxin B-1, sterigmatocystin, and terrein and compounds with chromophores of the shamixanthone, emerin and desertorin type of compounds. F. variecolor produces asteltoxin......, variecoxanthone A, B, C, isoemericellin, kojic acid, varitriol, varioxiran, dihydroterrein, 7-hydroxyemodin, avariquinone and stromemycin. E. pluriseminata produces several unknown specific extrolites. E. venezuelensis is the first organism of marine origin reported to produce aflatoxin. Aflatoxin production by E...

  14. Use of gamma irradiation to prevent aflatoxin B 1 production in smoked dried fish

    Science.gov (United States)

    Ogbadu, G. H.

    Smoked dried fish bought from the Nigerian market was inoculated with spores of barAspergillus flavus (U.I. 81) and irradiated with doses of 0.625, 1.25, 2.50 and 5.00 KGy gamma irradiation. The effect of aflatoxin B 1 production on subsequent incubation for 8 days as stationary cultures was measured. The amount of aflatoxin B 1 produced was found to decrease with increased gamma irradiation dose levels. While the non-irradiated control produced significantly (at 1% level) greater amounts of aflatoxin B 1 as compared to the treated cultures.

  15. Comparison of antibody production against aflatoxin B1 in goats and rabbits.

    OpenAIRE

    Gaur, P K; El-Nakib, O; Chu, F. S.

    1980-01-01

    Antibody production against aflatoxin B1 was compared in three rabbits and one goat. Titers obtained were 20 times higher in the rabbits than in the goat. The goat antiserum appeared to have a higher degree of cross-reactivity for other aflatoxins and related metabolites than did the rabbit antiserum.

  16. Aflatoxin B1 is toxic to porcine oocyte maturation.

    Science.gov (United States)

    Liu, Jun; Wang, Qiao-Chu; Han, Jun; Xiong, Bo; Sun, Shao-Chen

    2015-07-01

    As a toxic secondary metabolite of Aspergillus species, Aflatoxin B1 (AFB1) is a major food and feed contaminant in tropical and sub-tropical regions with high temperature and humidity. It has been reported to be toxic to the female reproductive system in laboratory and domestic animals. In the present study, the influence of acute exposure to AFB1 (10 and 50 μM, 44h) on porcine oocyte maturation and its possible mechanism were investigated. The maturation rates of oocytes decreased significantly in the presence of 50 μM of AFB1. Cell cycle analysis showed that most oocytes were arrested at germinal vesicle breakdown or meosis I stage. However, actin assembly, spindle structure and chromosome alignment were not disrupted after exposure to 50 μM AFB1. Further study showed that DNA methylation levels increased in treated oocytes (50 μM). Histone methylation levels were also analysed after treatment (50 μM): H3K27me3 and H3K4me2 levels decreased, whereas H3K9me3 level increased, indicating that epigenetic modification was affected. AFB1 treatment (50 μM) also induced oxidative stress and further led to autophagy, as shown by accumulation of reactive oxygen species, up-regulated LC3 protein expression and increased mRNA levels of ATG3, ATG5 and ATG7. Annexin V-FITC staining assay revealed that AFB1 treatment (50 μM) resulted in oocyte early apoptosis, which was confirmed by increased Bak, Bax, Bcl-xl mRNA levels. Collectively, our results suggest that AFB1 disrupts porcine oocyte maturation through changing epigenetic modifications as well as inducing oxidative stress, excessive autophagy and apoptosis.

  17. Aflatoxin B1 induced upregulation of protein arginine methyltransferase 5 in human cell lines.

    Science.gov (United States)

    Ghufran, Md Sajid; Ghosh, Krishna; Kanade, Santosh R

    2016-09-01

    The exposure of naturally occurring mycotoxins affects human health and play a vital role in cancer initiation and progression. Aflatoxin B1 is a difuranocoumarin mycotoxin, classified as a group I carcinogen. The present study was conducted to assess the effect of aflatoxin B1 on epigenetic regulatory proteins. The protein arginine methyltransferase 5 expression was induced upon aflatoxin B1 treatment in a dose and time dependent manner. Further global arginine methylation was also increased in the same manner. This is the first report showing the induction of epigenetic regulatory protein, protein arginine methyltransferase 5 upon aflatoxin B1 treatment. Further study is required to establish the detailed pathway of PRMT5 induction. PMID:27242039

  18. Distribution of [14C]-labelled aflatoxin B1 in mice

    International Nuclear Information System (INIS)

    The distribution of [14C]-labelled aflatoxin B1 has been studied in mice with the aid of whole-body autoradiography. In addition to the localisation of labelled aflatoxin B1 and/or its metabolites in the liver, bile, kidney, lung and urine an uptake of 14C in the pigment of the Harderian gland and the eye was observed. Uptake of radioactivity was also found in the eyes of the foetuses although their livers did not accumulate radioactivity. (author)

  19. Toxicity of aflatoxin B1 towards the vitamin D receptor (VDR).

    Science.gov (United States)

    Costanzo, Paola; Santini, Antonello; Fattore, Luigi; Novellino, Ettore; Ritieni, Alberto

    2015-02-01

    This research describes an unexpected toxicity of the aflatoxin B1 towards the vitamin D receptors. Rickets is a childhood disease, and calcium deficiency is the aetiological cause in Africa, being primarily associated with nutritional problems; in this research the contribution of aflatoxin B1 exposure during the early months of life is an interesting factor to deepen in order to prevent liver damages or the development of rickets. The results show that the expression of vitamin D receptor in osteosarcoma cell line SAOS-2 is significantly down-modulated by exposure to aflatoxin B1. This seems to suggest that Aflatoxin B1, toxic towards the vitamin D receptor, interferes with the actions of the vitamin D on calcium binding gene expression in the kidney and intestine. Experimental data indicate a 58% and 86% decrease if the cells are exposed to 5 ng/mL and 50 ng/mL of aflatoxin B1, respectively. These results seem to indicate that natural occurrence of the aflatoxin B1 and allelic variant of vitamin D receptor on (F allele) increase the risk of developing rickets of African children.

  20. Analysis of aflatoxins B1 and G1 in maize by quechers

    Directory of Open Access Journals (Sweden)

    Bursić Vojislava P.

    2013-01-01

    Full Text Available A reliable and easy method has been developed for the determination of aflatoxins B1 and G2 in maize samples. High performance liquid chromatography coupled with FLD (HPLC-FLD with photochemical derivatization was used. Mycotoxins were extracted from maize using a QuEChERS-based extraction procedure. The optimized analytical conditions were evaluated in terms of recoveries, reproducibility, LOD, LOQ and linearity for aflatoxin B1 and aflatoxin G1 in maize. Extraction, chromatographic and detection conditions were optimized in order to increase sample sensitivity. The linearity was analyzed in the range of 0.4-20 μg/kg and the correlation coefficients (R2 were higher than 0.99 for aflatoxins B1 and G1. Blank samples were spiked at 1.0, 2.0 and 4.0 μg/kg, and the average recovery for aflatoxin G1 was 96.96±1.72% and for aflatoxin B1 it was 86.80±1.24%. RSDs were lower than 25% for both mycotoxins. LOD for both aflatoxins was 0.5 μg/kg and LOQ was 1.0 μg/kg, respectively.

  1. Optimisation of Liquid Phase Separation on an Aflatoxin B1 Radioimmunoassay Kit

    International Nuclear Information System (INIS)

    Aflatoxins are secondary metabolites of Aspergillus flavus and A. parasiticus, which grow on a wide variety of food, feeds and their products. Aflatoxin, particularly aflatoxin B1 (AfB1) has wide biological activities including toxic, teratogenic, mutagenic and carcinogenic. Therefore the AfB1 content in foods should be met the requirement of food safety standard. Radioimmunoassay (RIA) technique is one of immunochemical methods or immunoassays which has been considered to be sensitive, specific, accurate and practical for detection of aflatoxin. This technique is based on immunological reaction using radioactive tracer. AfB1 was indirectly radiolabelled and then purified by using a solvent extraction. The tracer was then undergone a immunological testing by using a polyclonal antibody of AfB1. In order to produce AfB1 RIA kit which meets the standard quality’s requirement there are two parameters that have to be optimised. First is optimisation of kit components which consist : standard AfB1, AfB1 tracer (125I-AfB1) and AfB1 antibody. (author)

  2. Measurement and assessment of aflatoxin B1 and its producing molds in Iranian sausages and burgers

    Directory of Open Access Journals (Sweden)

    Siavash Maktabi

    2016-09-01

    Full Text Available Abstract Introduction: Aflatoxin B1 (AFB1 is one of the most well-known hepatocarcinogens in humans. Contamination of raw materials, used in the production of sausages and burgers, with aflatoxin producing molds can lead to increased level of aflatoxin in the final products and can impose hazards to human health. Unfortunately, aflatoxin is resistant to heating and freezing processes, etc. and can remain in these products untile consumption. Methods: During a six-month period, 45 sausage and 53 burger samples from valid brands across the country were randomly purchased from the stores. The samples were analyzed for AFB1 by ELISA technique. Meanwhile, the number of molds was calculated and aflatoxin producing molds were identified by direct and slide culture methods. Results: The findings showed that 2 susage samples (4.9% and 3 burger samples (6.3% were contaminated with >1 ng/g aflatoxin. Moreover, 4 burger samples (8.9% contaminated with mold included aspergillus flavus, aspergillus niger, mucor, and penicillium while, none of the susage samples showed mold contamination. Conclusion: The Iranian meat products had a relative aflatoxin B1 contamination during the study period, but the contamination rate was low and in allowable range. Standard hygienic preparation and packaging of meat products molds is recommended to reduce fungal contamination, especially aflatoxin-producing molds.

  3. Single corn kernel aflatoxin B1 extraction and analysis method

    Science.gov (United States)

    Aflatoxins are highly carcinogenic compounds produced by the fungus Aspergillus flavus. Aspergillus flavus is a phytopathogenic fungus that commonly infects crops such as cotton, peanuts, and maize. The goal was to design an effective sample preparation method and analysis for the extraction of afla...

  4. BIOCONTROL OF ASPERGILLUS FLAVUS AND AFLATOXIN B1 PRODUCTION IN CORN

    Directory of Open Access Journals (Sweden)

    A. Khanafari, H. Soudi, M. Miraboulfathi

    2007-07-01

    Full Text Available The potent mycotoxin aflatoxin B1 is a secondary metabolite of Aspergillus fungi that grow, on a variety of food and feed commodities at any stage during growth, harvest, storage and transportation. The occurrence of aflatoxin contamination is global, with severe problems especially prevalent in developing countries. In present study, corn samples were contaminated with aflatoxin B1 in the concentration of 240 µg/kg. Four trials were inoculated by Lactobacillus plantarum (PTCC 1058. Three control assays were analysed in the same conditions. All the assays were kneaded and incubated for 4-7 days at 37°C. Aflatoxin B1 was determined after extraction by HPLC. Results showed a drastic removal of the mycotoxin with a reduction of 77 % for Aflatoxin B1 by Lactobacillus plantarum. In the Inoculated corns, spore germination of A. flavus was totally inhibited. Results in inoculated spikes showed a high percentage of reduction of aflatoxin after incubation by Lb. plantarum. Gram staining of a sample from inoculated corns and microscopic observation demonstrated that the growth of A. flavus spores was totally inhibited by Lb. plantarum. Fungal spores were surrounded by Lactobacillus plantarum and spores were degraded.

  5. ESTIMATION OF AFLATOXIN B1 IN FEED INGREDIENTS AND COMPOUND POULTRY FEEDS

    Directory of Open Access Journals (Sweden)

    Bashir Mahmood Bhatti, Tanzeela Talat and Rozina Sardar

    2001-02-01

    Full Text Available A total of 3230 samples of feed ingredients of vegetable and animal origin and commercially available compound poultry feed received over a period of 5 years at Feed Testing Laboratory of the Institute were tested for Aflatoxin B1 contents (ppb . In all feed ingredients and compound feed stuffs, minimum level of aflatoxin B1 was 13 ppb and maximum level was found to be 78 ppb. No correlation of aflatoxin levels with month of collection of the year which are subject to variation in temperature and humidity could be detected. Mean values of aflatoxin concentration in feed stuffs such as rice, rice polish, wheat bran, wheat bread, maize, fish meal, blood meal, bone meal, guar meal, corn gluten 30%, corn gluten 60%, sun flower meal, soyabean meal and cotton seed meal were found to be higher than safe level of 20 ppb recommended by FDA.

  6. Moulds identification and detection of aflatoxin B1 on commercial codiments fermented of shrimp

    Directory of Open Access Journals (Sweden)

    NOOR SOESANTI HANDAJANI

    2006-07-01

    Full Text Available Indonesian tropical climate have an opportunity for fungi growth as Aspergillus flavus Link which can produce aflatoxin within foodstuffs, include condiment of fermented shrimp. Aflatoxin B1 is the dangerous agent having roles as carcinogenic, mutagenic and teratogenic. The aims of this research were known kinds of moulds and detection of aflatoxin B1 on commercial condiments fermented of shrimp. Two brands of commercial condiments fermented of shrimp were taken from traditional markets and supermarkets in Surakarta. Isolation was done by made suspension of sample in aquadets. Suspension on appropriate dilutions was grown on CDA (Czapek’s Dox Agar media with surface spread. The grown colonies were separated and grown on PDA (Potato Dextrose Agar slant media. Furthermore, isolates were cultured on CDA and MEA (Malt Extract Agar media. The grown colonies were microscopes and microscopes examined and identified. Existence of aflatoxin B1 was known by Commercial RIDA Screen ELISA Kit that could detect qualitatively and quantitatively with detection sensitive < 1.7 ppb. Moulds that could be isolated from condiments fermented of shrimp were: Aspergillus flavus Link, Aspergillus niger van Tieghem, Aspergillus wentii Wehmer, Aspergillus PU1 or Aspergillus PU2 and Penicillium citrinum Thom. There was aflatoxin B1 contaminated to 2 brands of commercial condiments fermented of shrimp that were examined. Traditional markets’ commercial condiments fermented of shrimp contained higher aflatoxin B1 than supermarkets’. The brands of commercial condiment of fermented shrimp which had better inner package quality contained lower aflatoxin B1 than the worst inner package quality of commercial condiments of fermented of shrimp.

  7. Effect of gamma radiation on the inactivation of aflatoxin B1 in food and feed crops

    International Nuclear Information System (INIS)

    Samples of food crops (peanut, peeled pistachio, unpeeled pistachio, rice, and corn) and feed (barley, bran, corn) were sterilized then inoculated with 106 of spore suspension of an isolate of Aspergillus flavus fungus known to produce aflatoxin B1 . Food and feed samples were irradiated with gamma radiation at the doses 4, 6, and 10 kGy. Results indicated that degradation of Aflatoxin B1 was positively correlated with the increase in the applied dose of gamma ray for each tested sample. For example, at a dose of 4 KGy. Percentages of aflatoxin B1 degradation were 8.4, 9.7, 16.6 and 23.5, and 43.97% for peanuts, peeled pistachios, unpeeled pistachios, corn and rice, consecutively . Whereas, at a dose of 10 KGy percentages of aflatoxin degradation reached highest values at 58.6, 68.8, 84.6, 81.1 and 87.8% for peanuts, peeled pistachios, unpeeled pistachios, corn and rice, consecutively In feed samples percentages of aflatoxin degradation were 45, 66, and 90% in barley, 47, 75, and 86% in bran, and 31, 72, and 84% in corn for the doses of 4, 6, and 10 KGy, consecutively. Aflatoxin degradation in food samples correlated negatively with oil content in irradiated samples. Thus, in peanuts, which contained the highest oil content, percentage of aflatoxin degradation at 10 KGy was not more than 56.6%, whereas, the corresponding value in corn, which contained the highest oil content, reached as high as 80%. The above results indicate the possibility of using gamma irradiation as a means of degradation of aflatoxin B1 in food and feed crops to lower than the maximum allowed levels using a maximum dose of radiation of 10 KGy which represents the permitted dose of radiation for such type of crops.(author)

  8. The aflatoxin B1 isolating potential of two lactic acid bacteria

    Institute of Scientific and Technical Information of China (English)

    Adel Hamidi; Reza Mirnejad; Emad Yahaghi; Vahid Behnod; Ali Mirhosseini; Sajad Amani; Sara Sattari; Ebrahim Khodaverdi Darian

    2013-01-01

    Objective:To determine lactic acid bacteria’s capability to enhance the process of binding and isolating aflatoxin B1 and to utilize such lactic acid bacteria as a food supplement or probiotic products for preventing absorption of aflatoxin B1 in human and animal bodies. Methods: In the present research, the bacteria were isolated from five different sources. For surveying the capability of the bacteria in isolating aflatoxin B1, ELISA method was implemented, and for identifying the resultant strains through 16S rRNA sequencing method, universal primers were applied. Results: Among the strains which were isolated, two strains of Lactobacillus pentosus and Lactobacillus beveris exhibited the capability of absorbing and isolating aflatoxin B1 by respectively absorbing and discharging 17.4%and 34.7%of the aforementioned toxin existing in the experiment solution. Conclusions:Strains of Lactobacillus pentosus and Lactobacillus beveris were isolated from human feces and local milk samples, respectively. And both strains has the ability to isolate or bind with aflatoxin B1.

  9. Rapid pretreatment and detection of trace aflatoxin B1 in traditional soybean sauce.

    Science.gov (United States)

    Xie, Fang; Lai, WeiHua; Saini, Jasdeep; Shan, Shan; Cui, Xi; Liu, DaoFeng

    2014-05-01

    Soybean sauce, a traditional fermented food in China, has different levels of aflatoxin B1 pollution. Two kinds of direct and indirect immunomagnetic bead methods for the pretreatment of aflatoxin B1 were evaluated in this work. A method was established to detect aflatoxin B1 in soybean sauce using an immunomagnetic bead system for pretreatment and ELISA for quantification. The pretreatment method of immunomagnetic beads performed better compared with the conventional extraction and immunoaffinity column method. ELISA exhibited a good linear relationship at an aflatoxin B1 concentration of 0.05-0.3μg/kg (r(2)=0.9842). The average recoveries across spike levels varied from 0.5 to 7μg/kg were 83.6-104% with a relative standard deviation between 4.2% and 11.7%. With the advantages of rapid detection, easy operation, simple equipment, sensitivity, accuracy, and high recovery; this method can be well applied in the trace determination of aflatoxin B1 in soybean sauce samples. PMID:24360425

  10. Inhibition effects of Chinese cabbage powder on aflatoxin B1-induced liver cancer.

    Science.gov (United States)

    Wang, Tuoyi; Li, Chunyan; Liu, Yang; Li, Tiezhu; Zhang, Jie; Sun, Yonghai

    2015-11-01

    In this study, 0.25 μg/ml aflatoxin B1 was used to establish a liver cancer model for assessing the potential anticancer ability of Chinese cabbage powder, which is a complex water-soluble extract from Chinese cabbage by spray-drying at an outlet temperature of 130 °C. We found at least 11 potential anticancer substances in Chinese cabbage powder. A 90-d animal experiment demonstrated that 10% of Chinese cabbage powder in drinking water could improve the plasma micronutrient status, inhibit the formation of aflatoxin B1-DNA adducts in liver cells, and effectively reduce the incidence of liver tumor induced by aflatoxin B1 from 6.67% to 0%. The dose effect experiment revealed that 10% may be the minimal effective dose to prevent the occurrence of early liver tumors. This study will help elucidate the basis of epidemiological observations of dietary cancer prevention in humans, as well as explore related mechanisms.

  11. Dietary modulation of the biotransformation and genotoxicity of aflatoxin B(1).

    Science.gov (United States)

    Gross-Steinmeyer, Kerstin; Eaton, David L

    2012-09-28

    Diet and its various components are consistently identified as among the most important 'risk factors' for cancer worldwide, yet great uncertainty remains regarding the relative contribution of nutritive (e.g., vitamins, calories) vs. non-nutritive (e.g., phytochemicals, fiber, contaminants) factors in both cancer induction and cancer prevention. Among the most potent known human dietary carcinogens is the mycotoxin, aflatoxin B(1) (AFB). AFB and related aflatoxins are produced as secondary metabolites by the molds Aspergillus flavus and Aspergillus parasiticus that commonly infect poorly stored foods including peanuts, pistachios, corn, and rice. AFB is a potent hepatocarcinogenic agent in numerous animal species, and has been implicated in the etiology of human hepatocellular carcinoma. Recent research has shown that many diet-derived factors have great potential to influence AFB biotransformation, and some efficiently protect from AFB-induced genotoxicity. One key mode of action for reducing AFB-induced carcinogenesis in experimental animals was shown to be the induction of detoxification enzymes such as certain glutathione-S-transferases that are regulated through the Keap1-Nrf2-ARE signaling pathway. Although initial studies utilized the dithiolthione drug, oltipraz, as a prototypical inducer of antioxidant response, dietary components such as suforaphane (SFN) are also effective inducers of this pathway in rodent models. However, human GSTs in general do not appear to be extensively induced by SFN, and GSTM1 - the only human GST with measurable catalytic activity toward aflatoxin B(1)-8,9-epoxide (AFBO; the genotoxic metabolite of AFB), does not appear to be induced by SFN, at least in human hepatocytes, even though its expression in human liver cells does appear to offer considerable protection against AFB-DNA damage. Although induction of detoxification pathways has served as the primary mechanistic focus of chemoprevention studies, protective effects of

  12. Dietary modulation of the biotransformation and genotoxicity of aflatoxin B1

    International Nuclear Information System (INIS)

    Diet and its various components are consistently identified as among the most important ‘risk factors’ for cancer worldwide, yet great uncertainty remains regarding the relative contribution of nutritive (e.g., vitamins, calories) vs. non-nutritive (e.g., phytochemicals, fiber, contaminants) factors in both cancer induction and cancer prevention. Among the most potent known human dietary carcinogens is the mycotoxin, aflatoxin B1 (AFB). AFB and related aflatoxins are produced as secondary metabolites by the molds Aspergillus flavus and Aspergillus parasiticus that commonly infect poorly stored foods including peanuts, pistachios, corn, and rice. AFB is a potent hepatocarcinogenic agent in numerous animal species, and has been implicated in the etiology of human hepatocellular carcinoma. Recent research has shown that many diet-derived factors have great potential to influence AFB biotransformation, and some efficiently protect from AFB-induced genotoxicity. One key mode of action for reducing AFB-induced carcinogenesis in experimental animals was shown to be the induction of detoxification enzymes such as certain glutathione-S-transferases that are regulated through the Keap1–Nrf2–ARE signaling pathway. Although initial studies utilized the dithiolthione drug, oltipraz, as a prototypical inducer of antioxidant response, dietary components such as suforaphane (SFN) are also effective inducers of this pathway in rodent models. However, human GSTs in general do not appear to be extensively induced by SFN, and GSTM1 – the only human GST with measurable catalytic activity toward aflatoxin B1-8,9-epoxide (AFBO; the genotoxic metabolite of AFB), does not appear to be induced by SFN, at least in human hepatocytes, even though its expression in human liver cells does appear to offer considerable protection against AFB–DNA damage. Although induction of detoxification pathways has served as the primary mechanistic focus of chemoprevention studies, protective

  13. Aflatoxin B1content in patients with hepatic diseases Aflatoxina B1 en pacientes con enfermedades hepáticas

    Directory of Open Access Journals (Sweden)

    Clara López

    2002-08-01

    Full Text Available Aflatoxins are toxic metabolites of some Aspergillus flavus, A. parasiticus and A. nomius strains that occur in many foods and feeds. There are four major natural occurring aflatoxins: B1, B2, G1 and G2. These toxins can cause illness in human beings and animals. Aflatoxin B1 is the most abundant and toxic member of the family, and it is also the most potent hepatocarcinogen known. In order to estimate the potential human health risk of AFB1, it is useful to measure blood concentration. The presence of aflatoxin B1 in patients was evaluated by high-performance liquid chromatography, in serum samples, obtained from 20 patient volunteers with hepatic disease. Out of the 20 patients, the presence of AFB1 was detected in only one of them, in a concentration of 0.47 ng/cm³. Nevertheless, this result should draw the attention of control organizations in Argentina to the need for a thorough food and feed inspection.Las aflatoxinas son metabolitos tóxicos producidos por cepas de Aspergillus flavus, A. parasiticus y A. nomius, presentes en alimentos y piensos. Las cuatro aflatoxinas principales son: aflatoxina B1, B2, G1 y G2. Dichas toxinas pueden causar enfermedades tanto en seres humanos como en animales. La aflatoxina B1 es la más abundante y la más tóxica del grupo y es también el más potente hepatocarcinógeno conocido. El objetivo de este trabajo fue detectar la presencia de aflatoxina B1 en sangre humana para estimar el riesgo potencial de la salud. La determinación de aflatoxina B1 fue realizada por cromatografía líquida de alto rendimiento, en suero de 20 pacientes voluntarios con enfermedades hepáticas. En sólo uno de estos pacientes se detectó la presencia de aflatoxina B1, en una concentración de 0.47ng/cm³. Estos resultados deberían ser tenidos en cuenta por los responsables de la vigilancia y control de los alimentos en la Argentina.

  14. Modulation of Aflatoxin B1 production by Aspergillus flavus

    OpenAIRE

    Verheecke, Carol

    2014-01-01

    Les mycotoxines sont des molécules toxiques produites par de nombreuses espèces fongiques. Les seules mycotoxines avérées aujourd’hui cancérigènes pour l’homme sont les aflatoxines. Elles sont produites par le genre Aspergillus principalement et sont retrouvées tout au long de la chaine alimentaire (champs, stockage, transformation, etc.). A cause du réchauffement climatique, la France devient de plus en plus exposée à la présence de ces mycotoxines. Afin de limiter l’exposition des consommat...

  15. The induction of neoplastic lesions by aflatoxin-B1 in the Egyptian toad (Bufo regularis).

    Science.gov (United States)

    el-Mofty, M M; Sakr, S A

    1988-01-01

    The carcinogenic activity of aflatoxin-B1, the metabolic product of the mold Aspergillus flavus (a commonly occurring contaminant of groundnuts and other foodstuffs), was tested using the Egyptian toad (Bufo regularis). Injecting the toads with aflatoxin-B1 at a dose level of 0.01 mg/50 g body wt in 1 ml corn oil once a week for 15 weeks induced hepatocellular carcinomas in 19% of the experimental toads. Four toads developed tumors in the kidney due to metastases from the primary hepatocellular carcinomas.

  16. Effect of climate change on Aspergillus flavus and aflatoxin B1 production

    Directory of Open Access Journals (Sweden)

    Angel eMedina

    2014-07-01

    Full Text Available This review considers the available information on the potential impact of key environmental factors and their interactions on the molecular ecology, growth and aflatoxin production by Aspergillus flavus in vitro and in maize grain. The recent studies which have been carried out to examine the impact of water activity x temperature on aflatoxin biosynthesis and phenotypic aflatoxin production are examined. These have shown that there is a direct relationship between the relative expression of key regulatory and structural genes under different environmental conditions which correlate directly with aflatoxin B1 production. A model has been developed to integrate the relative expression of 10 biosynthetic genes in the pathway, growth and aflatoxin B1 (AFB1 production which was validated under elevated temperature and water stress conditions. The effect of interacting conditions of aw x temperature x elevated CO2 (2x and 3x existing levels are detailed for the first time. This suggests that while such interacting environmental conditions have little effect on growth they do have a significant impact on aflatoxin biosynthetic gene expression (structural aflD and regulatory aflR genes and can significantly stimulate the production of AFB1. While the individual factors alone have an impact, it is the combined effect of these three abiotic factors which have an impact on mycotoxin production. This approach provides data which is necessary to help predict the real impacts of climate change on mycotoxigenic fungi.

  17. Effect of Gamma-irradiation on aflatoxin B1 produced by aspergillus parasiticus in barley containing antimicrobial food additives

    International Nuclear Information System (INIS)

    Influence of gamma irradiation on, growth and aflatoxin B1 produced by aspergillus parasiticus in ba supplemented with sodium chloride, potassium sorbate and sodium benzoate was investigated. Total viable population of A. Parasiticus and aflatoxin B1 production decreased significantly by increasing gamma irradiation doses. No growth or aflatoxin B1 production occurred at 4.0 KGy. Increasing the concentration of NaCl reduced the total viable population A. Parasiticus as well as the accumulation of aflatoxin B1. No growth and aflatoxin B1 production occurred in barley treated with 2.0 KGy and 6% NaCl. Potassium sorbate and sodium benzoate at concentration 500 ppm reduced the population of A. Parasiticus and the levels of aflatoxin B1 over 100 days. At 2.0 KGy, a sharp drop in aflatoxin B1 level occurred in barley by 2% NaCl and 500 ppm potassium sorbate and sodium benzoate. At 2.0 KGy, 2% NaCl and 1000 ppm potassium sorbate and sodium benzoate completely inhibited growth and aflatoxin B1 production by A. parasiticus for 100 days of incubation

  18. Genetic variation of Aflatoxin B(1) aldehyde reductase genes (AFAR) in human tumour cells

    DEFF Research Database (Denmark)

    Praml, Christian; Schulz, Wolfgang; Claas, Andreas;

    2008-01-01

    AFAR genes play a key role in the detoxification of the carcinogen Aflatoxin B(1) (AFB(1)). In the rat, Afar1 induction can prevent AFB(1)-induced liver cancer. It has been proposed that AFAR enzymes can metabolise endogenous diketones and dialdehydes that may be cytotoxic and/or genotoxic...

  19. [Growth and bioluminescence of luminous bacteria under the action of aflatoxin B1 before and after its treatment with nanodiamonds].

    Science.gov (United States)

    Mogil'naia, O A; Puzyr', A P; Bondar', V S

    2010-01-01

    The effect of aflatoxin B1 on growth and luminescence of marine luminous bacteria P. phosphoreum and recombinant E. coli Z905 cells was investigated. The bidirectional effect of aflatoxin B1 on the studied bacterial species was detected--an inhibition of luminescence in P. phosphoreum and its stimulation in E. coli. It was shown that aflatoxin B1 influences the cell luminescence in the freshly grown cultures and bacteria restored after lyophilization. It was detected that the effect of aflatoxin B1 was graded after interaction with the modified nanodiamond (MND) of detonation synthesis. After mycotoxin's treatment with MND, it does not cause significant changes in bacterial luminescence. The possibilities for the use of P. phosphoreum and E. coli bacteria in the bioluminescent monitoring of aflatoxin B1 and the use of MND for mycotoxin deactivation are discussed. PMID:20198915

  20. Transcriptomic Analysis of Aflatoxin B1-Regulated Genes in Rat Hepatic Epithelial Cells

    Institute of Scientific and Technical Information of China (English)

    杨柳; 季静; 李光辉; 李君文; 陈招立; 王海勇

    2014-01-01

    Aflatoxins are the most popular hepatotoxicants. Chronic exposure to aflatoxins leads to a wide variety of liver diseases, such as hepatocellular carcinoma. In this study, we analyzed the genome wide expression profiles of aflatoxin B1-induced rat hepatic epithelial cells. The expression of 325, 184 and 199 special genes was altered when exposed to 0.03, 0.1 and 0.2 μmol/L aflatoxin B1 respectively, and 239 genes were commonly expressed. After the functional analysis on these dose-special genes, we determined several key pathways related to hepatotoxicity, such as TGF-beta signaling pathway, tight junction, adherens junction, the regulation of actin cytoskeleton, ErbB signaling pathway, p53 signaling pathway, pathways in cancer and axon guidance. Common genes were mainly associated with focal adhesion, ECM-receptor interaction, and cell adhesion molecules. Gene ontology annotations showed a good concordance with these pathways. The quantitative real-time polymerase chain reaction(PCR) analysis of selected genes showed similar patterns in microarrays. The toxicogenomic study provides a better understanding of molecular mechanisms of aflatoxins.

  1. Acute toxicity of aflatoxin B1 and rubratoxin B in dogs.

    Science.gov (United States)

    Hayes, A W; Williams, W L

    1978-01-01

    The effect of ip administrated aflatoxin B1 and rubratoxin B, singly and in combination, on dogs was determined by serum tests, by observations of clinical signs and survival times, and by evaluation of gross and microscopic lesions. The dog is sensitive to the toxic effects of both mycotoxins. Glutamic-oxaloacetic transaminase, lactic dehydrogenase and alkaline phosphatase activities and survival time varied in relation to dose and to the mycotoxin(s) administered. All three plasma enzymes were elevated regardless of dose with the combination of aflatoxin B1/rubratoxin B at 24 hr after dosing, except LDH, which was within the normal range but only at the lowest dose level. Several serum constituents including BUN, cholesterol, uric acid, and total bilirubin were elevated, whereas serum glucose was depressed in dogs treated with the multiple-toxin regimen; these changes were not seen in dogs given only aflatoxin B1 but were characteristic in rubratoxin-treated animals. In general, gross findings at necropsy were similar in all dogs regardless of the dose regimen. A striking similarity existed in the histologic changes observed between lesions experimentally induced by the mycotoxin combination and those lesions reported for dogs fed toxic feed in laboratory studies or in natural cases of hepatitis X. Of particular similarity were the severe kidney lesions observed in dogs exposed to the mycotoxin combination and kidney lesions reported in natural outbreaks of hepatitis X. There can be little doubt of an association between hepatitis X and aflatoxin B1, although it is apparent that the disease probably involves more than a single toxic factor. Our results suggest that hepatitis X in dogs includes aflatoxin B1 as a primary etiological factor but that rubratoxin B also may be involved. PMID:581496

  2. Aflatoxin B1 contamination in maize in Europe increases due to climate change

    Science.gov (United States)

    Battilani, P.; Toscano, P.; Van der Fels-Klerx, H. J.; Moretti, A.; Camardo Leggieri, M.; Brera, C.; Rortais, A.; Goumperis, T.; Robinson, T.

    2016-01-01

    Climate change has been reported as a driver for emerging food and feed safety issues worldwide and its expected impact on the presence of mycotoxins in food and feed is of great concern. Aflatoxins have the highest acute and chronic toxicity of all mycotoxins; hence, the maximal concentration in agricultural food and feed products and their commodities is regulated worldwide. The possible change in patterns of aflatoxin occurrence in crops due to climate change is a matter of concern that may require anticipatory actions. The aim of this study was to predict aflatoxin contamination in maize and wheat crops, within the next 100 years, under a +2 °C and +5 °C climate change scenario, applying a modelling approach. Europe was virtually covered by a net, 50 × 50 km grids, identifying 2254 meshes with a central point each. Climate data were generated for each point, linked to predictive models and predictions were run consequently. Aflatoxin B1 is predicted to become a food safety issue in maize in Europe, especially in the +2 °C scenario, the most probable scenario of climate change expected for the next years. These results represent a supporting tool to reinforce aflatoxin management and to prevent human and animal exposure. PMID:27066906

  3. Aflatoxin B1 contamination in maize in Europe increases due to climate change

    Science.gov (United States)

    Battilani, P.; Toscano, P.; van der Fels-Klerx, H. J.; Moretti, A.; Camardo Leggieri, M.; Brera, C.; Rortais, A.; Goumperis, T.; Robinson, T.

    2016-04-01

    Climate change has been reported as a driver for emerging food and feed safety issues worldwide and its expected impact on the presence of mycotoxins in food and feed is of great concern. Aflatoxins have the highest acute and chronic toxicity of all mycotoxins; hence, the maximal concentration in agricultural food and feed products and their commodities is regulated worldwide. The possible change in patterns of aflatoxin occurrence in crops due to climate change is a matter of concern that may require anticipatory actions. The aim of this study was to predict aflatoxin contamination in maize and wheat crops, within the next 100 years, under a +2 °C and +5 °C climate change scenario, applying a modelling approach. Europe was virtually covered by a net, 50 × 50 km grids, identifying 2254 meshes with a central point each. Climate data were generated for each point, linked to predictive models and predictions were run consequently. Aflatoxin B1 is predicted to become a food safety issue in maize in Europe, especially in the +2 °C scenario, the most probable scenario of climate change expected for the next years. These results represent a supporting tool to reinforce aflatoxin management and to prevent human and animal exposure.

  4. Assessment of aflatoxin B1 in livestock feed and feed ingredients by high-performance thin layer chromatography

    Directory of Open Access Journals (Sweden)

    Korrapati Kotinagu

    2015-12-01

    Full Text Available Aim: Detection of aflatoxin B1 in Livestock compound Feed and feed ingredients by high-performance thin layer chromatography (HPTLC. Materials and Methods: Chromatography was performed on HPTLC silica gel 60 F 254, aluminum sheets by CAMAG automatic TLC sampler 4, with mobile phase condition chloroform:acetone:water (28:4:0.06. Extraction of aflatoxin B1 from samples was done as per AOAC method and screening and quantification done by HPTLC Scanner 4 under wavelength 366 nm. Results: A total of 97 livestock feed (48 and feed ingredients (49 samples received from different livestock farms and farmers were analyzed for aflatoxin B1of which 29 samples were contaminated, constituting 30%. Out of 48 livestock compound feed samples, aflatoxin B1 could be detected in 16 samples representing 33%, whereas in livestock feed ingredients out of 49 samples, 13 found positive for aflatoxin B1 representing 24.5%. Conclusion: HPTLC assures good recovery, precision, and linearity in the quantitative determination of aflatoxin B1 extracted from Livestock compound feed and feed ingredients. As more number of feed and feed ingredients are contaminated with aflatoxin B1 which causes deleterious effects in both animal and human beings, so there is a need for identifying the source of contamination, executing control measures, enabling better risk assessment techniques, and providing economic benefits.

  5. Effect of dietary acids on the formation of aflatoxin B2a as a means to detoxify aflatoxin B1.

    Science.gov (United States)

    Rushing, Blake R; Selim, Mustafa I

    2016-09-01

    Aflatoxin B1 (AFB1) is a class 1 carcinogen and a common food contaminant worldwide with widely uncontrolled human exposure. The ability of organic acids to transform AFB1 into a known detoxified form, aflatoxin B2a (AFB2a), was investigated using high performance liquid chromatography-electrospray ionisation-time of flight mass spectrometry (HPLC/ESI/TOF/MS). The identity of the transformation product was confirmed by accurate mass measurement, chromatographic separation from other aflatoxins, H(1)-nuclear magnetic resonance (NMR) and infrared (IR) spectroscopy. Of the weak acids tested, citric acid was found to be the most effective for AFB2a formation. At room temperature, 1 M citric acid was able to convert > 97% of AFB1 to AFB2a over 96 h of treatment. Up to 98% transformation was achieved by boiling AFB1 in the presence of citric acid for 20 min. AFB1 hydration after ingestion was explored by spiking AFB1 into simulated gastric fluid containing citric acid. Under these conditions, > 71% of AFB1 was hydrated to AFB2a and did not show any reversion to the parent compound after being transferred to a neutral solution. These results provide a basis for a practical and effective method for detoxification of AFB1 in contaminated foods.

  6. Effect of dietary acids on the formation of aflatoxin B2a as a means to detoxify aflatoxin B1.

    Science.gov (United States)

    Rushing, Blake R; Selim, Mustafa I

    2016-09-01

    Aflatoxin B1 (AFB1) is a class 1 carcinogen and a common food contaminant worldwide with widely uncontrolled human exposure. The ability of organic acids to transform AFB1 into a known detoxified form, aflatoxin B2a (AFB2a), was investigated using high performance liquid chromatography-electrospray ionisation-time of flight mass spectrometry (HPLC/ESI/TOF/MS). The identity of the transformation product was confirmed by accurate mass measurement, chromatographic separation from other aflatoxins, H(1)-nuclear magnetic resonance (NMR) and infrared (IR) spectroscopy. Of the weak acids tested, citric acid was found to be the most effective for AFB2a formation. At room temperature, 1 M citric acid was able to convert > 97% of AFB1 to AFB2a over 96 h of treatment. Up to 98% transformation was achieved by boiling AFB1 in the presence of citric acid for 20 min. AFB1 hydration after ingestion was explored by spiking AFB1 into simulated gastric fluid containing citric acid. Under these conditions, > 71% of AFB1 was hydrated to AFB2a and did not show any reversion to the parent compound after being transferred to a neutral solution. These results provide a basis for a practical and effective method for detoxification of AFB1 in contaminated foods. PMID:27467853

  7. Aflatoxin B1-producing Aspergillus in sun-dried medicinal plant materials

    OpenAIRE

    Chinaputi, A.; Lim, S; Petcharat, V.; Chuenchit, S.; Pathanadech, A.

    2001-01-01

    Fifty sun-dried medicinal plants were obtained from fraditional drug stores in Songkhla Province, Thailand, and examined for Aspergillus and aflatoxin B1. 288 isolates of Aspergillus were obtaines by standard blotter plate and 25 species were identified. The most common species were A. niger with 99 isolates, A. Flavus 84 isolates, A. terreus 33 isolates, A. oryzae 25 isolates, A.nidulans (Emericella nidulans) 10 isolates, A fumigatus 9 isolates and A. chevalieri (Eurotium chevalieri) 8 isola...

  8. Indirect determination of aflatoxin B1 in beer by a multicommuted optical sensor

    OpenAIRE

    Molina García, Lucía Molina; Fernández De Córdova, M. Luisa; Ruiz Medina, Antonio

    2012-01-01

    Abstract This manuscript reports the determination of Aflatoxin B1 (AFB1), mycotoxin which is considered one of the most carcinogenic substances known. A multicommuted flow injection-solid phase spectroscopy (FI-SPS) system combined with photochemically-induced fluorescence (PIF) is developed, for the first time, for its determination with quantitative purposes. A strongly fluorescent degradation product is obtained on-line by irradiation with ultraviolet light. The determination i...

  9. Effect of phenolic antioxidants on the mutagenicity of aflatoxin B1.

    OpenAIRE

    Shelef, L. A.; Chin, B

    1980-01-01

    The Ames assay employing Salmonella typhimurium TA100 and TA98 was used to investigate potential interactions between aflatoxin B1 (AFB1) and the phenolic antioxidants butylated hydroxytoluene, butylated hydroxyanisole, and propyl gallate. AFB1 doses were within the linear response range, and the antioxidants were used at levels of 0 to 50 micrograms per plate. All three antioxidants were nonmutagenic in either bacterial tester strain, with or without the hepatic S-9 enzyme preparation; toxic...

  10. Biosensor-based screening method for the detection of aflatoxins B1-G1.

    Science.gov (United States)

    Cuccioloni, Massimiliano; Mozzicafreddo, Matteo; Barocci, Simone; Ciuti, Francesca; Pecorelli, Ivan; Eleuteri, Anna Maria; Spina, Michele; Fioretti, Evandro; Angeletti, Mauro

    2008-12-01

    Aflatoxins are extremely toxic metabolites from Aspergillus species that can adulterate a wide range of human foodstuff. Herein, we propose a novel assay designed as an analytical test for aflatoxin B1 and G1 (AFB1 and AFG1, respectively) that could represent an alternative screening technique for this class of mycotoxins. The approach for the determination of these toxins is based on surface plasmon resonance using neutrophil porcine elastase as a "bait" for these aflatoxins. The selection and optimization of the analytical procedure involved a preliminary investigation on the type of inhibition by AFB1: the level of the protease inhibition exerted by AFB1 depended upon the incubation time and the concentration of the binding partners, showing the competitiveness and the reversibility of the inhibition. A posteriori, the nature of the interaction granted a rapid analysis, a single detection test requiring only a few minutes. For the development of the assay, the experimental conditions were evaluated and optimized with both calibration solution and aflatoxin-spiked samples. To apply this method to aflatoxin-contaminated maize, a rapid solid-phase extraction treatment was developed. The proposed assay for AFB1 and AFG1 was validated by comparison with both a chromatographic reference method and a standard enzyme linked immunosorbent assay procedure. This enzyme-based biosensor represents a new approach for the detection of aflatoxins based on the reversible interaction between a blocked macromolecule and a soluble ligand, having the major advantages in the relative rapidity, the reusability of the capturing surface, and low cost per single test. PMID:19551989

  11. Simultaneous detection of cyclopiazonic acid and aflatoxin B1 by HPLC in methanol/water mobile phase

    OpenAIRE

    Soares, Célia Maria Gonçalves; Freitas-Silva, O.; Abrunhosa, Luís; Venâncio, Armando

    2009-01-01

    A simple procedure for the simultaneous detection of cyclopiazonic acid (CPA) and aflatoxin B1 from fungal extracts is presented, using a methanol and water mobile phase and fluorescence detection. This methodology has been tested with standard solutions of both mycotoxins CPA and Aflatoxin B1 and with methanolic extracts of Aspergillus section Flavi strains, previously characterized for their mycotoxin production profile. Previously available methodology required the use of two different c...

  12. Assessment of aflatoxin B1 in livestock feed and feed ingredients by high-performance thin layer chromatography

    OpenAIRE

    Korrapati Kotinagu; T. Mohanamba; L. Rathna Kumari

    2015-01-01

    Aim: Detection of aflatoxin B1 in Livestock compound Feed and feed ingredients by high-performance thin layer chromatography (HPTLC). Materials and Methods: Chromatography was performed on HPTLC silica gel 60 F 254, aluminum sheets by CAMAG automatic TLC sampler 4, with mobile phase condition chloroform:acetone:water (28:4:0.06). Extraction of aflatoxin B1 from samples was done as per AOAC method and screening and quantification done by HPTLC Scanner 4 under wavelength 366 nm. Results: ...

  13. A Theoretical Study of 8-Chloro-9-Hydroxy-Aflatoxin B1, the Conversion Product of Aflatoxin B1 by Neutral Electrolyzed Water

    Science.gov (United States)

    Escobedo-González, René; Méndez-Albores, Abraham; Villarreal-Barajas, Tania; Aceves-Hernández, Juan Manuel; Miranda-Ruvalcaba, René; Nicolás-Vázquez, Inés

    2016-01-01

    Theoretical studies of 8-chloro-9-hydroxy-aflatoxin B1 (2) were carried out by Density Functional Theory (DFT). This molecule is the reaction product of the treatment of aflatoxin B1 (1) with hypochlorous acid, from neutral electrolyzed water. Determination of the structural, electronic and spectroscopic properties of the reaction product allowed its theoretical characterization. In order to elucidate the formation process of 2, two reaction pathways were evaluated—the first one considering only ionic species (Cl+ and OH−) and the second one taking into account the entire hypochlorous acid molecule (HOCl). Both pathways were studied theoretically in gas and solution phases. In the first suggested pathway, the reaction involves the addition of chlorenium ion to 1 forming a non-classic carbocation assisted by anchimeric effect of the nearest aromatic system, and then a nucleophilic attack to the intermediate by the hydroxide ion. In the second studied pathway, as a first step, the attack of the double bond from the furanic moiety of 1 to the hypochlorous acid is considered, accomplishing the same non-classical carbocation, and again in the second step, a nucleophilic attack by the hydroxide ion. In order to validate both reaction pathways, the atomic charges, the highest occupied molecular orbital and the lowest unoccupied molecular orbital were obtained for both substrate and product. The corresponding data imply that the C9 atom is the more suitable site of the substrate to interact with the hydroxide ion. It was demonstrated by theoretical calculations that a vicinal and anti chlorohydrin is produced in the terminal furan ring. Data of the studied compound indicate an important reduction in the cytotoxic and genotoxic potential of the target molecule, as demonstrated previously by our research group using different in vitro assays. PMID:27455324

  14. A Theoretical Study of 8-Chloro-9-Hydroxy-Aflatoxin B1, the Conversion Product of Aflatoxin B1 by Neutral Electrolyzed Water

    Directory of Open Access Journals (Sweden)

    René Escobedo-González

    2016-07-01

    Full Text Available Theoretical studies of 8-chloro-9-hydroxy-aflatoxin B1 (2 were carried out by Density Functional Theory (DFT. This molecule is the reaction product of the treatment of aflatoxin B1 (1 with hypochlorous acid, from neutral electrolyzed water. Determination of the structural, electronic and spectroscopic properties of the reaction product allowed its theoretical characterization. In order to elucidate the formation process of 2, two reaction pathways were evaluated—the first one considering only ionic species (Cl+ and OH− and the second one taking into account the entire hypochlorous acid molecule (HOCl. Both pathways were studied theoretically in gas and solution phases. In the first suggested pathway, the reaction involves the addition of chlorenium ion to 1 forming a non-classic carbocation assisted by anchimeric effect of the nearest aromatic system, and then a nucleophilic attack to the intermediate by the hydroxide ion. In the second studied pathway, as a first step, the attack of the double bond from the furanic moiety of 1 to the hypochlorous acid is considered, accomplishing the same non-classical carbocation, and again in the second step, a nucleophilic attack by the hydroxide ion. In order to validate both reaction pathways, the atomic charges, the highest occupied molecular orbital and the lowest unoccupied molecular orbital were obtained for both substrate and product. The corresponding data imply that the C9 atom is the more suitable site of the substrate to interact with the hydroxide ion. It was demonstrated by theoretical calculations that a vicinal and anti chlorohydrin is produced in the terminal furan ring. Data of the studied compound indicate an important reduction in the cytotoxic and genotoxic potential of the target molecule, as demonstrated previously by our research group using different in vitro assays.

  15. Five of 12 forms of vaccinia virus-expressed human hepatic cytochrome P450 metabolically activate aflatoxin B1

    International Nuclear Information System (INIS)

    Twelve forms of human hepatic cytochrome P450 were expressed in hepatoma cells by means of recombinant vaccinia viruses. The expressed P450s were analyzed for their abilities to activate the potent hepatocarcinogen aflatoxin B1 to metabolites having mutagenic or DNA-binding properties. Five forms, P450s IA2, IIA3, IIB7, IIIA3, and IIIA4, activated aflatoxin B1 to mutagenic metabolites as assessed by the production of His revertants of Salmonella typhimurium in the Ames test. The same P450s catalyzed conversion of aflatoxin B1 to DNA-bound derivatives as judged by an in situ assay in which the radiolabeled carcinogen was incubated with cells expressing the individual P450 forms. Seven other human P450s, IIC8, IIC9, IID6, IIE1, IIF1, and IIIA5, and IVB1, did not significantly activate aflatoxin B1 as measured by both the Ames test and the DNA-binding assay. Moreover, polyclonal anti-rat liver P450 antibodies that crossreact with individual human P450s IA2, IIA3, IIIA3, and IIIA4 each inhibited aflatoxin B1 activation catalyzed by human liver S-9 extracts. Inhibition ranged from as low as 10% with antibody against IIA3 to as high as 65% with antibody against IIIA3 and IIIA4. These results establish that metabolic activation of aflatoxin B1 in human liver involves the contribution of multiple forms of P450

  16. Cytochrome P3-450 cDNA encodes aflatoxin B1-4-hydroxylase.

    Science.gov (United States)

    Faletto, M B; Koser, P L; Battula, N; Townsend, G K; Maccubbin, A E; Gelboin, H V; Gurtoo, H L

    1988-09-01

    Aflatoxin B1 (AFB1), a potent hepatocarcinogen and ubiquitous dietary contaminant in some countries, is detoxified to aflatoxin M1 (AFM1) via cytochrome P-450-mediated AFB1-4-hydroxylase. Genetic studies in mice have demonstrated that the expression of AFB1-4-hydroxylase is regulated by the aryl hydrocarbon locus and suggested that different cytochrome P-450 isozymes catalyze AFB1-4-hydroxylase and aryl hydrocarbon hydroxylase activities. We have now examined lysates from mammalian cells infected with recombinant vaccinia viruses containing expressible cytochrome P1-450 or P3-450 cDNAs for their ability to metabolize AFB1 to AFM1. Our results show that cytochrome P3-450 cDNA specifies AFB1-4-hydroxylase. This is the first direct assignment of a specific cytochrome P-450 to an AFB1 detoxification pathway. This finding may have relevance to the dietary modulation of AFB1 hepatocarcinogenesis.

  17. Determination and chemometric evaluation of total aflatoxin, aflatoxin B1, ochratoxin A and heavy metals content in corn flours from Turkey.

    Science.gov (United States)

    Algül, Işıl; Kara, Derya

    2014-08-15

    Concentrations of the total aflatoxin, aflatoxin B1, aflatoxin B2, aflatoxin G1, aflatoxin G2, ochratoxin A, lead, cadmium, mercury, arsenic, copper, zinc and chromium in corn flour samples were determined. Eighteen corn flour samples that were obtained from different cities and villages in Turkey and 3 corn flour samples obtained from the UK. Determination of the different toxins was carried out using HPLC instrumentation after pre-separation using immunoaffinity columns that work through a mechanism of solid-phase extraction. An ICP-MS instrument was used for the heavy metal determinations. The results obtained from HPLC and ICP-MS analyses of the corn flour samples showed that these samples contain detectable levels of most of the analytes but the mercury was at undetectable levels. A very strong statistical relationship was observed between Cr and total Aflatoxin and Aflatoxin B1; whereas Ochratoxin A was related to Cu and Zn concentrations using correlation analyses and principal component analyses. PMID:24679753

  18. CARRY-OVER OF AFLATOXIN B1-FEED INTO AFLATOXIN M1-MILK IN DAIRY COWS TREATED WITH NATURAL SOURCES OF AFLATOXIN AND BENTONITE

    Directory of Open Access Journals (Sweden)

    I. Sumantri

    2012-12-01

    Full Text Available High occurrence of aflatoxin contamination in feed stuffs implicates for a long time experience of aflatoxin B1 (AFB1 exposure to dairy cattle in Indonesia. A latin square 4X4 research design was adopted to study the characteristic of AFB1 carry-over rate (COR of Indonesian crossbred Friesian Holstein (PFH as effects of inclusions of AFB1-naturally contaminated feed and bentonite in the diet. Results showed a rapid aflatoxin M1 (AFM1 excretion in the milk, detected in the first milking sample or 10 hours after AFB1 ingestion. The steady state of AFM1 concentration observed since the first day of treatment period and AFM1 contamination was still detected until 5 days after AFB1 removed from the diet. The COR in this study was observed 0.1%. AFM1 concentration was highly significantly (P0.05 of levels of AFB1 and bentonite inclusions on the COR, nutrients intake, milk production, and milk composition. IIt is concluded that AFM1 concentration was influenced by AFB1 intake and that transfer of AFB1-feed into AFM1-milk (COR in PFH cow was lower compare to reported COR value for dairy cow in sub tropical region.

  19. Transformation of adsorbed aflatoxin B1 on smectite at elevated temperatures

    Science.gov (United States)

    Aflatoxins cause liver damage and suppress immunity. Smectites can be used to reduce the bioavailability of aflatoxins through adsorption. To further reduce the toxicity of aflatoxins and to eliminate the treatments of aflatoxin-loaded smectites, degrading the adsorbed aflatoxin to nontoxic or less ...

  20. Aflatoxin B1 levels in groundnut and sunflower oils in different Sudanese states.

    Science.gov (United States)

    Mariod, Abdalbasit Adam; Idris, Yousif Mohamed Ahmed

    2015-01-01

    In this article, the level of contamination of aflatoxin B1 (AFB1) in groundnut and sunflower oils was determined. The 241 oil samples were collected from Khartoum, Gezira, Kordofan and Algadarif states of Sudan and assessed for AFB1 using high-performance liquid chromatography (HPLC). AFB1 levels in groundnut oil samples ranged from 0.5 to 70 µg/kg and were 0.7 to 35 µg/kg in sunflower oil samples. High contamination was found in unrefined samples. It was concluded that AFB1 levels in oil samples indicated that growing, harvesting, handling and storage of the crops were not done properly.

  1. Determination of Aflatoxin B1 Levels in Organic Spices and Herbs

    OpenAIRE

    Halil Tosun; Recep Arslan

    2013-01-01

    Organically produced spices and herbs were analyzed for determination of aflatoxin B1 (AFB1) by ELISA using immunoaffinity column. For this purpose 93 organic spices and 37 organic herbs were randomly selected from organic markets and organic shops in Turkey. AFB1 was detected in 58 organic spice and 32 organic herb samples. Among organic spice samples, the maximum value was detected in cinnamon sample (53  μ g/kg). AFB1 was not detected in thyme samples. AFB1 levels of 41 organic spice sampl...

  2. Prevention of Aflatoxin B1-Induced DNA Breaks by β-D-Glucan

    Directory of Open Access Journals (Sweden)

    Eduardo Madrigal-Bujaidar

    2015-06-01

    Full Text Available Aflatoxins are a group of naturally-occurring carcinogens that are known to contaminate different human and animal foodstuffs. Aflatoxin B1 (AFB1 is the most genotoxic hepatocarcinogenic compound of all of the aflatoxins. In this report, we explore the capacity of β-D-glucan (Glu to reduce the DNA damage induced by AFB1 in mouse hepatocytes. For this purpose, we applied the comet assay to groups of animals that were first administered Glu in three doses (100, 400 and 700 mg/kg bw, respectively and, 20 min later, 1.0 mg/kg of AFB1. Liver cells were obtained at 4, 10 and 16 h after the chemical administration and examined. The results showed no protection of the damage induced by AFB1 with the low dose of the polysaccharide, but they did reveal antigenotoxic activity exerted by the two high doses. In addition, we induced a co-crystallization between both compounds, determined their fusion points and analyzed the molecules by UV spectroscopy. The data suggested the formation of a supramolecular complex between AFB1 and β-D-glucan.

  3. Effects of a Calcium Bentonite Clay in Diets Containing Aflatoxin when Measuring Liver Residues of Aflatoxin B1 in Starter Broiler Chicks

    Directory of Open Access Journals (Sweden)

    Justin Fowler

    2015-08-01

    Full Text Available Research has shown success using clay-based binders to adsorb aflatoxin in animal feeds; however, no adsorbent has been approved for the prevention or treatment of aflatoxicosis. In this study, growth and relative organ weights were evaluated along with a residue analysis for aflatoxin B1 in liver tissue collected from broiler chickens consuming dietary aflatoxin (0, 600, 1200, and 1800 µg/kg both with and without 0.2% of a calcium bentonite clay additive (TX4. After one week, only the combined measure of a broiler productivity index was significantly affected by 1800 µg/kg aflatoxin. However, once birds had consumed treatment diets for two weeks, body weights and relative kidney weights were affected by the lowest concentration. Then, during the third week, body weights, feed conversion, and the productivity index were affected by the 600 µg/kg level. Results also showed that 0.2% TX4 was effective at reducing the accumulation of aflatoxin B1 residues in the liver and improving livability in birds fed aflatoxin. The time required to clear all residues from the liver was less than one week. With evidence that the liver’s ability to process aflatoxin becomes relatively efficient within three weeks, this would imply that an alternative strategy for handling aflatoxin contamination in feed could be to allow a short, punctuated exposure to a higher level, so long as that exposure is followed by at least a week of a withdrawal period on a clean diet free of aflatoxin.

  4. Aflatoxin B1 binding capacity of autochthonous strains of lactic acid bacteria.

    Science.gov (United States)

    Fazeli, Mohammad R; Hajimohammadali, M; Moshkani, Azamossadat; Samadi, Nasrin; Jamalifar, Hossein; Khoshayand, Mohammad R; Vaghari, Elham; Pouragahi, Samieh

    2009-01-01

    Some foods are prone to contamination with aflatoxins, with detrimental effect on human health. Lactic acid bacteria have been reported to bind aflatoxins and remove them from foods and feeds. Reduction of aflatoxin B1 (AFB1) from the liquid media by the autochthonous lactic acid bacteria (Lactobacillus casei, Lactobacillus plantarum, and Lactobacillus fermentum) isolated from traditional Iranian sourdough and dairy products is reported in the current study. The effect of incubation time on the binding capacity of the strains to AFB1 was also investigated. Duplicates of individual bacteria with population equivalent to 2 X 10(10) CFU/ml were incubated in the presence of AFB1 at 37 degrees C for a period of 72 h, and the amounts of unbound AFB1 were quantitated by reverse-phase high-performance liquid chromatography. All the strains were capable of removal of AFB1, and the reduction of AFB1 ranged from 25 to 61% throughout the incubation period. Removal of AFB1 was a rapid process, with approximately 61 and 56% of the toxin taken instantly by L. fermentum and L. plantarum, respectively. Binding was of a reversible nature, and some of the bound AFB1 was released into the media by the repeated centrifugation and resuspension of the cell pellets. The stability of the bacteria-toxin complex was strain dependent, and L. casei was a stronger binder of AFB1 compared with the other bacteria. No toxin release was observed after 24 h. These findings tend to suggest that certain novel probiotic bacteria with high aflatoxin binding capacity could be selected for detoxification of foods. PMID:19205485

  5. Affinity maturation of single-chain variable fragment specific for aflatoxin B(1) using yeast surface display.

    Science.gov (United States)

    Min, Won-Ki; Kim, Sung-Gun; Seo, Jin-Ho

    2015-12-01

    As aflatoxin B1 is one of the most toxic mycotoxins, it is important to detect and to quantify aflatoxin B1 accurately by immunological methods. To enhance aflatoxin B1-binding affinity of the single-chain variable fragment, yeast surface display technique combined with fluorescence-activated cell sorting was applied. A randomly mutated scFv library was subjected to 4 rounds of fluorescence-activated cell sorting, resulting in isolation of 5 scFv variants showing an affinity improvement compared to the parental wild type scFv. The best scFv with a 9-fold improvement in affinity for aflatoxin B1 exhibited similar specificity to the monoclonal antibody. Most of the mutations in scFv-M37 were located outside of the canonical antigen-contact loops, suggesting that its affinity improvement might be driven by an allosteric effect inducing scFv-M37 to form a more favorable binding pocket for aflatoxin B1 than the wild type scFv. PMID:26041237

  6. Protection against aflatoxin-B1-induced liver mutagenesis by Scutellaria baicalensis.

    Science.gov (United States)

    de Boer, Johan G; Quiney, Brendan; Walter, Patrick B; Thomas, Cynthia; Hodgson, Kimberley; Murch, Susan J; Saxena, Praveen K

    2005-10-15

    We have measured the inhibition of the mutagenicity of the mycotoxin aflatoxin-B(1) in the liver of the rat by plant material of Scutellaria baicalensis, or Huang-qin. The addition of one percent dried Huang-qin to the feed of the animals reduced the mutant frequency of a subsequent administration of aflatoxin-B1 by approximately 60 and 77%, respectively, for two different batches of the plant material. The addition of Huang-qin also increased the expression of the gene for glutathione S-transferase A5 subunit by 2.5-3.0-fold, and decreased expression of P450 cytochrome 3A2 by 1.8-2.0-fold. The greater increase of the expression of the GST gene may result in the protection shown by Huang-qin. The sensitivity of the hepatic mitochondria to swelling, a measure of the mitochondrial permeability transition, is increased significantly in animals that are on a diet containing Huang-qin. This may lead to increased sensitivity to apoptosis on treatment with toxic compounds. The two batches of Huang-qin material show differences in both chemical composition and preventive potential. This study demonstrates how a combination of generating and analysis of plant varieties together with a mammalian assay for efficacy may improve the search for better plant-based prevention of cancer initiation. PMID:16202794

  7. Potential natural exposure of Mississippi sandhill cranes to aflatoxin B1.

    Science.gov (United States)

    Couvillion, C E; Jackson, J R; Ingram, R P; Bennett, L W; McCoy, C P

    1991-10-01

    A survey was conducted to determine if carcinogenic mycotoxins were present in foods consumed by Mississippi sandhill cranes (Grus canadensis pulla). Samples of field corn (Zea mays) (n = 111) and chufa (Cyperus esculentus) (n = 20), obtained in 1987, 1988 and 1989 on the Mississippi Sandhill Crane National Wildlife Refuge (MSCNWR) and nearby private lands were analyzed for aflatoxin B1(AB1), ochratoxin A and sterigmatocystin using thin layer chromatography. Chufa samples were negative for all three mycotoxins. Aflatoxin B1 was found in corn at concentrations from 5 to 5,000 ppb; the other mycotoxins were not found in corn. Contaminated corn was found in 72% of all corn fields, but the proportion of contaminated fields was 57 to 100% for the 3-yr period. Contamination with AB1 was greatest in corn obtained from the ground post-harvest. Overall, 32% of corn samples from the ground had levels greater than or equal to 200 ppb with a mean of 427 ppb (range = 5 to 5,000 ppb) in contaminated fields. In 1989, mean AB1 concentration in corn on the ground was 5 to 1138 ppb for individual fields. The concentration of AB1 was less than or equal to 200 ppb in all corn samples from upright stalks. The study demonstrated that AB1 is available to sandhill cranes and at levels that may pose a serious health threat. PMID:1758031

  8. Effect of water molecules on the fluorescence enhancement of Aflatoxin B1 mediated by Aflatoxin B1:beta-cyclodextrin complexes. A theoretical study.

    Science.gov (United States)

    Ramírez-Galicia, Guillermo; Garduño-Juárez, Ramón; Gabriela Vargas, M

    2007-01-01

    In order to explain the observed fluorescence enhancement of Aflatoxin B1 (AFB1) when forming AFB1:beta-cyclodextrin (AFB1:beta-CD) inclusion complexes, we have performed a theoretical (quantum chemistry calculations) study of AFB1 and AFB1:beta-CD in vacuum and in the presence of aqueous solvent. The AM1 method was used to calculate the absorption and emission wavelengths of these molecules. With the help of density functional theory (DFT) and time-dependent DFT (TDDFT) vibrational frequencies and related excitation energies of AFB1 and AFB1.(H2O)m = 4,5,6,11 were calculated. On the basis of these calculations we propose a plausible mechanism for the fluorescence enhancement of AFB1 in the presence of beta-CD: (1) before photoexcitation of AFB1 to its S1 excited state, there is a vibrational coupling between the vibrational modes involving the AFB1 carbonyl groups and the bending modes of the nearby water molecules (CG + WM); (2) these interactions allow a thermal relaxation of the excited AFB1 molecules that results in fluorescence quenching; (3) when the AFB1 molecules form inclusion complexes with beta-CD the CG + WM interaction decreases; and (4) this gives rise to a fluorescence enhancement.

  9. A Gold Nanoparticle and Aflatoxin B1-BSA Conjugates Based Lateral Flow Assay Method for the Analysis of Aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Sangdae Lee

    2012-04-01

    Full Text Available A rapid and simple immuno-chromatographic assay was developed to detect aflatoxin B1 (AFB1. The assay was based on a modified competitive binding format using colloidal gold and polyclonal antibody (Pab conjugates. The anti-AFB1 Pab was immobilized to a defined detection zone on a porous nitrocellulose membrane and colloidal gold particles were conjugated to AFB1-BSA which served as a detection reagent. The AFB1-containing sample was added to the membrane and allowed to move with AFB1-BSA-coated particles dried on the conjugation pad. The mixture was then passed along the porous membrane by capillary action past the Pab in the detection zone, which captured AFB1 or AFB1-BSA. AFB1 in the sample inhibits binding of AFB1-BSA conjugated gold particles to the Pab and prevents formation of a red color dot. In the absence of AFB1, AFB1-BSA conjugated gold particles bound to the Pab, give a red color within this detection zone. With this method, 10 μg/mL of AFB1 was detected in less than 10 min. The developed AFB1 assay also showed no cross reaction to Ochratoxin A (OTA.

  10. A preliminary study on the physiological effects of aflatoxin B-1 lactating water buffaloes

    International Nuclear Information System (INIS)

    Aflatoxin B-1 is one of the biologically active mycotoxins, produced as contaminants in human and animal food by a variety of spoilage molds. The effects of administering aflatoxin B-1 on plasma proteins, total thyroxine, cholesterol, zinc and iron were studied in eight lactating buffaloes (3rd and 8th season of lactation). The toxin was first tested in one animal which received single doses of 400, 1000 and 1500 ug each, mixed with 1 Kg ration, given one week apart. Blood, milk and urine samples were collected over the next 120 hrs. The toxin was not detected in either milk or urine. One week later, the latter dose was offered daily, but the animal lost its appetite and did not consume any of the ration offered on the second day. On the third day through the sixth, the toxin (1500 ug in 5 ml chloroform) was injected into the rumen through the flank region using a long needle and a syringe. However, the toxin could not be detected in either milk or urine. After another week, the dose was increased to 5000 ug intraruminally given for 2 days. The toxin appeared in milk and urine. The other seven animals were then included in the experiment and blood samples were collected 10 and 34 hours after dosing. The results obtained (from the pilot test) showed a transient decrease in serum albumin which lasted for 10-24 hours after oral administration of 400, 1000 and 1500 ug in food. This phenomenon was also confirmed in all animals at 34 hrs. after the intraruminal administration of 5000 ug (p>0.01). On the other hand, serum cholesterol was increased (P>0.01). Serum zinc was also increased though insignificantly. However, no appreciable changes were noted in either serum iron or total thyroxine. This experiment has shown that physiological responses may occur before the detection of the toxin in body fluids. It is suggested that measurement of plasma proteins and cholesterol can be used as a test for the toxic effect of aflatoxin, especially at low doses; a case similar to

  11. Evidence of mycotoxins (aflatoxin B1 and ochratoxin A) using the radioimmunoassay (RIA) in naturally contaminated cereals

    International Nuclear Information System (INIS)

    The aim of our study was to gain starting information on the aflatoxin B1 and ochratoxin A levels in cereals and feed mixtures which are in poultry breeding. To ascertain the presence of mycotoxins, we examined the cultivars of cereals (maize and wheat) and the feed mixtures. The cereals came from different regions of eastern Slovakia. In all cereals examined, the low mycotoxin levels did not exceed the tolerance limit set by hygienic standard (Sv. 61, 1986, No. 69). In wheat, the contamination by aflatoxin B1 ranged from 0.028 to 0.125 μg·kg-1. In maize, the contamination by aflatoxin B1 ranged from 0.166 to 0.707 μg·kg-1. The results enhance our knowledge of feedstuff and feed mixture contamination in poultry breeding

  12. CARRY-OVER OF AFLATOXIN B1-FEED INTO AFLATOXIN M1-MILK IN DAIRY COWS TREATED WITH NATURAL SOURCES OF AFLATOXIN AND BENTONITE

    Directory of Open Access Journals (Sweden)

    I. Sumantri

    2014-10-01

    Full Text Available High occurrence of aflatoxin contamination in feed stuffs implicates for a long time experience ofaflatoxin B1 (AFB1 exposure to dairy cattle in Indonesia. A latin square 4X4 research design wasadopted to study the characteristic of AFB1 carry-over rate (COR of Indonesian crossbred FriesianHolstein (PFH as effects of inclusions of AFB1-naturally contaminated feed and bentonite in the diet.Results showed a rapid aflatoxin M1 (AFM1 excretion in the milk, detected in the first milking sampleor 10 hours after AFB1 ingestion. The steady state of AFM1 concentration observed since the first dayof treatment period and AFM1 contamination was still detected until 5 days after AFB1 removed fromthe diet. The COR in this study was observed 0.1%. AFM1 concentration was highly significantly(P<0.01 affected by treatments. However, there were no significant effects (P>0.05 of levels of AFB1and bentonite inclusions on the COR, nutrients intake, milk production, and milk composition. IIt isconcluded that AFM1 concentration was influenced by AFB1 intake and that transfer of AFB1-feed intoAFM1-milk (COR in PFH cow was lower compare to reported COR value for dairy cow in sub tropicalregion.

  13. [Exposure to aflatoxin B1 in experimental animals and its public health significance].

    Science.gov (United States)

    Guzmán de Peña, Doralinda

    2007-01-01

    The presence of AFB1 in human beings was detected in Mexico in 1996 both as a mutation of the gene p53 in hepatocellular carcinomas in Monterrey, Mexico, and as the adduct AFB1-lysine in serum from patients in Matamoros, Mexico in 2003. Aflatoxin B1 has been classified as a carcinogenic agent to humans by the International Agency for Research on Cancer. The compound is a natural contaminant produced by Aspergillus flavus and/or A. parasiticus when these fungi grow on different food products. At the molecular level, this review covers the carcinogenic, mutagenic and toxic properties of these mycotoxins and their risk to humans. It also gives insight into the causal relationship between aflatoxins and hepatocellular carcinoma. Information is provided about AFB1-formamidopyrimidine, which is a determinant of the carcinogenic and mutagenic capabilities. The results suggest that the Mexican population ingests food containing low amounts of AFB1. Analyses is presented of AFB1 toxicity, which is a consequence of the carcinogenic activity in liver cells.

  14. Occurrence of Aspergillus spp. and aflatoxin B1 in Malaysian foods used for human consumption.

    Science.gov (United States)

    Reddy, Kasa R N; Farhana, Nazira I; Salleh, Baharuddin

    2011-05-01

    Malaysian population widely consumes the cereal-based foods, oilseeds, nuts, and spices in their daily diet. Mycotoxigenic fungi are well known to invade food products under storage conditions and produce mycotoxins that have threat to human and animal health. Therefore, determining toxigenic fungi and aflatoxin B(1) (AFB1) in foods used for human consumption is of prime importance to develop suitable management strategies and to minimize risk. Ninety-five food products marketed in Penang, Malaysia were randomly collected from different supermarkets and were analyzed for presence of Aspergillus spp. by agar plate assay and AFB1 by enzyme-linked immunosorbent assay (ELISA). A. flavus was the dominant fungi in all foods followed by A. niger. Fifty-five A. flavus strains were tested for their ability to produce aflatoxins on rice grain substrate. Thirty-six (65.4%) strains out of 55 produced AFB1 ranging from 1700 to 4400 μg/kg and 17 strains (31%) produced AFB2 ranging from 620 to 1670 μg/kg. Natural occurrence of AFB1 could be detected in 72.6% food products ranging from 0.54 to 15.33 μg/kg with a mean of 1.95 μg/kg. Maximum AFB1 levels were detected in peanut products ranging from 1.47 to 15.33 μg/kg. AFB1 levels detected in all food products were below the Malaysian permissible limits (Malaysia.

  15. Effect of three different anti-mycotoxin additives on broiler chickens exposed to aflatoxin B1

    Directory of Open Access Journals (Sweden)

    AA Oliveira

    2015-01-01

    Full Text Available The growth of filamentous fungi on food often causes, aside from its deterioration, the mycotoxin production which determines economic losses in poultry industry, such as decreased productivity and injuries on poultry's carcass. Adsorbents based on yeast cell wall from Saccharomyces cerevisiae, which contain esterified glucomannan, are an alternative to reduce the mycotoxins bioavailability. The aim of this study was to compare in vitro and in vivo the performance of new three anti-mycotoxin additives (AMA based on yeast cell wall from Saccharomyces cerevisiae. The adsorption process was quantified in vitro, and the data obtained when plotted with Hill's equation indicated a cooperative process. Then, the three different AMA were tested for its ability to reduce the effects of aflatoxins in the diet of growing broiler chickens. The addition of 1 mg kg-1 aflatoxin B1 to the diets of broilers caused a negative change on the performance parameters besides increasing liver weight, fatty degeneration and liver necrosis. The addition of two different kinds of AMA (0.2% could reverse such effects. In conclusion, AMA 1 and 2 are additives with good potential for application on animal production. The AMA 3 ingredients must be re-tested alone for its adsorption capacity. These are the first data reported from Brazil anti-mycotoxin additives with preliminary isothermal analysis. Since beneficial characteristics of S. cerevisiae cell wall in animal industry are strain dependent, this study suggests two new promising alternatives to ameliorate mycotoxin problem.

  16. Exposure to aflatoxin B1 in late gestation alters protein kinase C and apoptotic protein expression in murine placenta.

    Science.gov (United States)

    Wang, Yanfei; Tan, Wenjuan; Wang, C C; Leung, Lai K

    2016-06-01

    Mycotoxins are chemicals with diverse toxicities that are produced by fungi. Aflatoxin B1 is commonly found in plant food, and is generally regarded as one of the most toxic mycotoxins. In the present study, pregnant ICR mice were given p.o. daily doses of aflatoxin B1 at 0, 0.05, 0.5, 5mg/kg for 4days (from E13.5 to E16.5). Compared to the control group, time of delivery was shortened and low birth weight was induced in mice treated with 0.5 and 5mg aflatoxin B1/kg, respectively. Placental tissue isolated from pregnant mice at E17.5 showed that the mRNA expression of crh was increased in aflatoxin-treated groups. This upregulation might signify premature delivery. Further analysis indicated that Pkc proteins were activated and Bcl-2 was reduced in the placental tissue of the aflatoxin-treated groups. Reduction of the anti-apoptotic proteins, on the other hand, might affect the morphorgenesis and maintenance of the placenta. PMID:26968497

  17. Aflatoxin B1, zearalenone and deoxynivalenol in feed ingredients and complete feed from central China.

    Science.gov (United States)

    Liu, Jie; Sun, Lvhui; Zhang, Jiacai; Guo, Jiao; Chen, Lei; Qi, Desheng; Zhang, Niya

    2016-06-01

    Between 2012 and 2014, 2528 feed ingredient and complete feed samples were collected from central China. Numbers of 2083, 255 and 190 samples were analysed for aflatoxin B1 (AFB1), zearalenone (ZEN) and deoxynivalenol (DON), respectively, by high-performance liquid chromatography in combination with UV or fluorescence detection. The incidence rates of AFB1, ZEN and DON contamination of feed ingredients and complete feeds were 33.9%, 90.2% and 77.4%, respectively. The percentage of positive samples for AFB1 ranged from 13.1% to 97.1%. Cottonseed meal presented the most serious contamination by AFB1. ZEN and DON contamination levels of feeds ranged from 50% to 100%, indicating serious contamination over the studied 3-year period. This study demonstrates that AFB1, ZEN and DON contamination of feeds in central China is serious and differs over the years. Feeds are mostly contaminated with ZEN, followed by DON and AFB1. PMID:26771914

  18. Prenatal exposure to aflatoxin B1: developmental, behavioral, and reproductive alterations in male rats

    Science.gov (United States)

    Supriya, Ch.; Reddy, P. Sreenivasula

    2015-06-01

    Previous studies have shown that aflatoxin B1 (AfB1) inhibits androgen biosynthesis as a result of its ability to form a high-affinity complex with the steroidogenic acute regulatory protein. The results of the present study demonstrate the postnatal effects of in utero exposure to AfB1 in the rat. Pregnant Wistar rats were given 10, 20, or 50 μg AfB1/kg body weight daily from gestation day (GD) 12 to GD 19. At parturition, newborns were observed for clinical signs and survival. All animals were born alive and initially appeared to be active. Male pups from control and AfB1-exposed animals were weaned and maintained up to postnatal day (PD) 100. Litter size, birth weight, sex ratio, survival rate, and crown-rump length of the pups were significantly decreased in AfB1-exposed rats when compared to controls. Elapsed time (days) for testes to descend into the scrotal sac was significantly delayed in experimental pups when compared to control pups. Behavioral observations such as cliff avoidance, negative geotaxis, surface rightening activity, ascending wire mesh, open field behavior, and exploratory and locomotory activities were significantly impaired in experimental pups. Body weights and the indices of testis, cauda epididymis, prostate, seminal vesicles, and liver were significantly reduced on PD 100 in male rats exposed to AfB1 during embryonic development when compared with controls. Significant reduction in the testicular daily sperm production, epididymal sperm count, and number of viable, motile, and hypo-osmotic tail coiled sperm was observed in experimental rats. The levels of serum testosterone and activity levels of testicular hydroxysteroid dehydrogenases were significantly decreased in a dose-dependent manner with a significant increase in the serum follicle-stimulating hormone and luteinizing hormone in experimental rats. Deterioration in the testicular and cauda epididymal architecture was observed in experimental rats. The results of fertility

  19. Optical waveguide lightmode spectroscopy technique-based immunosensor development for aflatoxin B1 determination in spice paprika samples.

    Science.gov (United States)

    Majer-Baranyi, Krisztina; Zalán, Zsolt; Mörtl, Mária; Juracsek, Judit; Szendrő, István; Székács, András; Adányi, Nóra

    2016-11-15

    Optical waveguide lightmode spectroscopy (OWLS) technique has been applied to label-free detection of aflatoxin B1 in a competitive immunoassay format, with the aim to compare the analytical goodness of the developed OWLS immunosenor with HPLC and enzyme-linked immunosorbent assay (ELISA) methods for the detection of aflatoxin in spice paprika matrix. We have also assessed applicability of the QuEChERS method prior to ELISA measurements, and the results were compared to those obtained by traditional solvent extraction followed by immunoaffinity clean-up. The AFB1 content of sixty commercial spice paprika samples from different countries were measured with the developed and optimized OWLS immunosensor. Comparing the results from the indirect immunosensor to that obtained by HPLC or ELISA provided excellent correlation (with regression coefficients above 0.94) indicating that the competitive OWLS immunosensor has a potential for quick determination of aflatoxin B1 in paprika samples. PMID:27283719

  20. Effect of curcumin on hepatic antioxidant enzymes activities and gene expressions in rats intoxicated with aflatoxin B1.

    Science.gov (United States)

    El-Bahr, S M

    2015-01-01

    Twenty-eight rats were examined in a 5-week experiment to investigate the effect of curcumin on gene expression and activities of hepatic antioxidant enzymes in rats intoxicated with aflatoxin B1 (AFB1 ). The rats were divided into four groups. Rats in 1-4 groups served as control, oral curcumin treated (15 mg/kg body weight), single i.p. dose of AFB1 (3 mg/kg body weight) and combination of single i.p. dose of AFB1 with oral curcumin treated, respectively. AFB1 Liver damage and oxidative stress were evident in untreated AFB1 -intoxicated rats as indicated by a significant elevation in hepatic transaminases, elevation in lipid peroxide biomarkers (thiobarbituric acid reactive substances; TBARS), reduction of reduced glutathione (GSH) concentration, reduction in the activities of antioxidant enzymes namely catalase (CAT), total superoxide dismutase (SOD), glutathione peroxidase (GPX) and glutathione-S-transferase (GST) and down-regulation of gene expression of these antioxidant enzymes compared to control. Liver sections of rats intoxicated with AFB1 showed a disrupted lobular architecture, scattered necrotic cells and biliary proliferation. Administration of curcumin with AFB1 resulted in amelioration of AFB1 -induced effects compared to untreated AFB1 -intoxicated rats via an up-regulation of antioxidant enzyme gene expression, activation of the expressed genes and increase in the availability of GSH.

  1. Risk Assessment on Dietary Exposure to Aflatoxin B1 in Post-Harvest Peanuts in the Yangtze River Ecological Region

    Directory of Open Access Journals (Sweden)

    Xiaoxia Ding

    2015-10-01

    Full Text Available Based on the 2983 peanut samples from 122 counties in six provinces of China’s Yangtze River ecological region collected between 2009–2014, along with the dietary consumption data in Chinese resident nutrition and health survey reports from 2002 and 2004, dietary aflatoxin exposure and percentiles in the corresponding statistics were calculated by non-parametric probability assessment, Monte Carlo simulation and bootstrap sampling methods. Average climatic conditions in the Yangtze River ecological region were calculated based on the data from 118 weather stations via the Thiessen polygon method. The survey results found that the aflatoxin contamination of peanuts was significantly high in 2013. The determination coefficient (R2 of multiple regression reflected by the aflatoxin B1 content with average precipitation and mean temperature in different periods showed that climatic conditions one month before harvest had the strongest impact on aflatoxin B1 contamination, and that Hunan and Jiangxi provinces were greatly influenced. The simulated mean aflatoxin B1 intake from peanuts at the mean peanut consumption level was 0.777–0.790 and 0.343–0.349 ng/(kg·d for children aged 2–6 and standard adults respectively. Moreover, the evaluated cancer risks were 0.024 and 0.011/(100,000 persons·year respectively, generally less than China’s current liver cancer incidence of 24.6 cases/(100,000 persons·year. In general, the dietary risk caused by peanut production and harvest was low. Further studies would focus on the impacts of peanut circulation and storage on aflatoxin B1 contamination risk assessment in order to protect peanut consumers’ safety and boost international trade.

  2. Co-mutagenicity of coumarin (1,2-benzopyrone) with aflatoxin B1 and human liver S9 in mammalian cells.

    Science.gov (United States)

    Goeger, D E; Hsie, A W; Anderson, K E

    1999-06-01

    Coumarin (1,2-benzopyrone), a natural dietary constituent and drug currently under evaluation for treatment of certain cancers and lymphedema, reduces polycyclic aromatic hydrocarbon-induced neoplasms in rodents. Because most rodents metabolize coumarin through 3,4-epoxidation, whereas 7-hydroxylation predominates in humans, their suitability as a model for coumarin effects in humans has been questioned. We examined coumarin chemoprotection against the promutagen and dietary contaminant aflatoxin B1 with human liver S9 bioactivation in the Chinese hamster ovary cell/hypoxanthine-guanine phosphoribosyltransferase mutation assay. Coumarin in the absence of aflatoxin B1 was not mutagenic or cytotoxic up to 500 microM. When included with either 1 or 10 microM aflatoxin B1, coumarin produced a dose-dependent increase in mutant frequency and cytotoxicity. At concentrations greater than 50 microM, coumarin stimulated human liver S9 bioactivation of aflatoxin B1 to the mutagenic 8,9-epoxide. This increase was 12- and fivefold at 500 microM coumarin with 1 and 10 microM aflatoxin B1, respectively, compared with incubations with aflatoxin B1 alone. These findings differ from previous results with liver S9 from other species, and indicate that coumarin co-mutagenicity with aflatoxin B1 and human liver S9 is through increased aflatoxin B1 bioactivation.

  3. Effect of Plectranthus glandulosus and Ocimum gratissimum Essential Oils on Growth of Aspergillus flavus and Aflatoxin B1 Production

    Directory of Open Access Journals (Sweden)

    Mbofung, CMF.

    2008-01-01

    Full Text Available Essential oils of Ocimum gratissimum and Plectranthus glandulosus leaves were extracted by steam distillation and analysed by GC-MS, and their effects on growth and aflatoxin B1 production by Aspergillus flavus were tested at five levels (i.e 200, 400, 600, 800 and 1000 mg/l using SMKY agar medium. The main components of O. gratissimum were thymol (47.7% and -terpinene (14.3% whereas those of P. glandulosus were represented by -terpinene (30.8% and terpinolene (25.2%. After 8 days of incubation on essential oil-supplemented medium, growth of A. flavus was totally inhibited by 800 mg/l of O. gratissimum essential oil and by 1000 mg/l of P. glandulosus essential oil. The effect of essential oils on aflatoxin B1 synthesis was evaluated in SMKY broth. The medium supplemented with different essential oil concentrations, was inoculated with A. flavus mycelium and incubated at 25 °C. At 2, 4, 6 and 8 days, aflatoxin B1 concentrations in the supernatant were estimated using Enzyme Linked Immuno-Sorbent Assay (ELISA. Results showed that aflatoxin B1 synthesis was inhibited by 1000 mg/l of both essential oils of O. gratissimum and P. glandulosus after 8 days of incubation. Results obtained in the present study indicate the possibility of exploiting O. gratissimum and P. glandulosus essential oils in the fight against strains of A. flavus responsible for biodeterioration of stored food products.

  4. Effect of aflatoxin B1 on in vitro ruminal fermentation of rations high in alfalfa hay or ryegrass hay

    DEFF Research Database (Denmark)

    Jiang, Y H; Yang, H J; Lund, Peter

    2012-01-01

    A 2 × 4 factorial experiment was conducted to determine the effect of aflatoxin B1 (AFB1) at dose rates of 0, 320, 640, 960 ng/ml on ruminal fermentation of substrates high in alfalfa hay (HA, alfalfa hay: maize meal = 4:1) and ryegrass hay (HR, ryegrass hay: maize meal = 4:1). In vitro dry matter...

  5. Aflatoxin B(1) in affecting broiler's performance, immunity, and gastrointestinal tract: a review of history and contemporary issues.

    Science.gov (United States)

    Yunus, Agha W; Razzazi-Fazeli, E; Bohm, Josef

    2011-06-01

    Aflatoxin B(1) is a common contaminant of poultry feeds in tropical and subtropical climates. Research during the last five decades has well established the negative effects of the mycotoxin on health of poultry. However, the last ten years of relevant data have accentuated the potential of low levels of aflatoxin B(1) to deteriorate broiler performance. In this regard, any attempt to establish a dose-effect relationship between aflatoxin B(1) level and broiler performance is also complicated due to differences in types of broilers and length of exposure to the mycotoxin in different studies. Contrary to the prevalent notion regarding literature saturation with respect to aflatoxicosis of chicken, many areas of aflatoxicosis still need to be explored. Literature regarding effects of the mycotoxin on the gastrointestinal tract in this regard is particular scanty and non-conclusive. In addition to these issues, the metabolism of aflatoxin B(1) and recently proposed hypotheses regarding biphasic effects of the mycotoxin in broilers are briefly discussed.

  6. Determination of urinary biomarkers for assessment of short-term human exposure to aflatoxins in São Paulo, Brazil.

    Science.gov (United States)

    Jager, Alessandra V; Tonin, Fernando G; Souto, Pollyana C M C; Privatti, Rafaela T; Oliveira, Carlos A F

    2014-07-08

    In the present study, a longitudinal assessment was carried out to evaluate the short-term human exposure to aflatoxins in Pirassununga region, São Paulo, Brazil, by determination of urinary aflatoxins by a liquid chromatography coupled to mass sprectrometry (UPLC-MS/MS) method. Sixteen volunteers with ages ranging from 14 to 55 years old were instructed to collect the early morning first urine four times every three months, from June 2011 to March 2012, totaling 64 samples. Aflatoxin M1 (AFM1) was found in 39 samples (61%) at levels ranging from 0.19 to 12.7 pg·mg-1 creatinine (mean: 1.2 ± 2.0 pg·mg-1 creatinine). Residues of aflatoxins B1, B2, G1, G2 and aflatoxicol were not identified in any urine sample. No significant difference was found among the AFM1 mean levels in urine samples collected in the four sampling periods. The levels of AFM1 found in urine samples indicate a low short-term exposure of the population studied to aflatoxins through the diet, although further investigations are needed to assess other long-term biomarkers of exposure to AFB1.

  7. Hepatoprotective Role of Milk Thistle (Silybum marianum in Meat Type Chicken Fed Aflatoxin B1 Contaminated Feed

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    Din Muhammad, Naila Chand, Sarzamin Khan*, Asad Sultan, Mohammad Mushtaq and Rafiullah

    2012-06-01

    Full Text Available Milk thistle was added in aflatoxin B1 contaminated poultry feed to investigate and compare its hepatoprotective effects with a commercial toxin binder. Two hundred and forty, day-old broilers were randomly allocated into four major groups A, B, C and D. Group A was kept as control, having aflatoxin free feed, while group B was fed aflatoxin contaminated feed, group C was raised on aflatoxin contaminated feed with toxin binder “Mycoad” @ 3g/kg of feed, while group D was provided aflatoxin contaminated feed along with milk thistle @10g/kg of feed. Aflatoxin B1 was present at the level of 80 µg/kg feed during the first week and 520 µg/kg feed in the remaining experimental period. Serum total protein was significantly (P<0.05 higher in group D, followed by group A, C and B. Serum enzymes including, alkaline phosphatase (ALP, aspartate aminotransferase (AST and alanine aminotransferase (ALT values were significantly (P<0.05 lower in group D, followed by C, A and B, which are indicative of hepatoprotective role of milk thistle. Body weight gain and feed intake was decreased by aflatoxin contaminated feed (group B in comparison with group A and group D. Milk thistle supplementation improved body weight gain and feed intake and was similar to toxin binder treated birds. Average feed conversion ratio (FCR was significantly (P<0.05 higher (poor in group B and were the same in all other groups. Present study demonstrated that milk thistle can potentially be used as mycotoxin binder and to minimize the adverse effects of toxin contaminated feed in broilers production.

  8. Impairment of cell cycle progression by aflatoxin B1 in human cell lines.

    Science.gov (United States)

    Ricordy, R; Gensabella, G; Cacci, E; Augusti-Tocco, G

    2002-05-01

    Aflatoxin B1 is a mycotoxin produced by Aspergillus flavus and Aspergillus parasiticum, which may be present as a food contaminant. It is known to cause acute toxic effects and act as a carcinogenic agent. The carcinogenic action has been related to its ability to form unstable adducts with DNA, which represent possible mutagenic sites. On the other hand, the primary cellular target responsible for its toxic action has not yet been clearly identified. Previous data suggested a possible correlation between cell proliferation and responsiveness to aflatoxin toxicity. These observations led us to investigate the effect of the toxin on cell cycle progression of three human cell lines (HepG2, SK-N-MC and SK-N-SH derived from liver and nervous tissue tumours); they were shown to display different responses to toxin exposure and have different growth kinetics. We performed analysis of the cell cycle, DNA synthesis and expression of p21 and p53 in the presence and absence of the toxin in all cell lines exposed. The results of cell cycle cytofluorometric analysis show significant alterations of cell cycle progression as a result of toxin treatment. In all cell lines exposure to a 24 h toxin treatment causes a dose-dependent accumulation in S phase, however, the ability to recover from impairment to traverse S phase varies in the cell lines under study. SK-N-MC cells appear more prone to resume DNA synthesis when the toxin is removed, while the other two cell lines maintain a significant inhibition of DNA synthesis, as indicated by cytofluorimetry and [(3)H]dTR incorporation. The level of p53 and p21 expression in the three cell lines was examined by western blot analysis and significant differences were detected. The ready resumption of DNA synthesis displayed by SK-N-MC cells could possibly be related to the absence of p53 control of cell cycle progression.

  9. Aflatoxin B1 degradation by liquid cultures and lysates of three bacterial strains.

    Science.gov (United States)

    Adebo, Oluwafemi Ayodeji; Njobeh, Patrick Berka; Sidu, Sibusiso; Tlou, Matsobane Godfrey; Mavumengwana, Vuyo

    2016-09-16

    Aflatoxin contamination remains a daunting issue to address in food safety. In spite of the efforts geared towards prevention and elimination of this toxin, it still persists in agricultural commodities. This has necessitated the search for other measures such as microbial degradation to combat this hazard. In this study, we investigated the biodegradation of aflatoxin B1 (AFB1), using lysates of three bacterial strains (Pseudomonas anguilliseptica VGF1, Pseudomonas fluorescens and Staphylococcus sp. VGF2) isolated from a gold mine aquifer. The bacterial cells were intermittently lysed in the presence and absence of protease inhibitors to obtain protease free lysates, subsequently incubated with AFB1 for 3, 6, 12, 24, and 48h to investigate whether any possible AFB1 degradation occurred using high performance liquid chromatography (HPLC) for detection. Results obtained revealed that after 6h of incubation, protease inhibited lysates of Staphylococcus sp. VGF2 demonstrated the highest degradation capacity of 100%, whereas P. anguilliseptica VGF1 and P. fluorescens lysates degraded AFB1 by 66.5 and 63%, respectively. After further incubation to 12h, no residual AFB1 was detected for all the lysates. Lower degrading ability was however observed for liquid cultures and uninhibited lysates. Data on cytotoxicity studies against human lymphocytes showed that the degraded products were less toxic than the parent AFB1. From this study, it can thus be deduced that the mechanism of degradation by these bacterial lysates is enzymatic. This study shows the efficacy of crude bacterial lysates for detoxifying AFB1 indicating potential for application in the food and feed industry. PMID:27294556

  10. Aflatoxin B1 and M1 Degradation by Lac2 from Pleurotus pulmonarius and Redox Mediators

    Science.gov (United States)

    Loi, Martina; Fanelli, Francesca; Zucca, Paolo; Liuzzi, Vania C.; Quintieri, Laura; Cimmarusti, Maria T.; Monaci, Linda; Haidukowski, Miriam; Logrieco, Antonio F.; Sanjust, Enrico; Mulè, Giuseppina

    2016-01-01

    Laccases (LCs) are multicopper oxidases that find application as versatile biocatalysts for the green bioremediation of environmental pollutants and xenobiotics. In this study we elucidate the degrading activity of Lac2 pure enzyme form Pleurotus pulmonarius towards aflatoxin B1 (AFB1) and M1 (AFM1). LC enzyme was purified using three chromatographic steps and identified as Lac2 through zymogram and LC-MS/MS. The degradation assays were performed in vitro at 25 °C for 72 h in buffer solution. AFB1 degradation by Lac2 direct oxidation was 23%. Toxin degradation was also investigated in the presence of three redox mediators, (2,2′-azino-bis-[3-ethylbenzothiazoline-6-sulfonic acid]) (ABTS) and two naturally-occurring phenols, acetosyringone (AS) and syringaldehyde (SA). The direct effect of the enzyme and the mediated action of Lac2 with redox mediators univocally proved the correlation between Lac2 activity and aflatoxins degradation. The degradation of AFB1 was enhanced by the addition of all mediators at 10 mM, with AS being the most effective (90% of degradation). AFM1 was completely degraded by Lac2 with all mediators at 10 mM. The novelty of this study relies on the identification of a pure enzyme as capable of degrading AFB1 and, for the first time, AFM1, and on the evidence that the mechanism of an effective degradation occurs via the mediation of natural phenolic compounds. These results opened new perspective for Lac2 application in the food and feed supply chains as a biotransforming agent of AFB1 and AFM1. PMID:27563923

  11. Chemopreventive effect of cactus Opuntia ficus indica on oxidative stress and genotoxicity of aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Ben Mansour Hédi

    2011-10-01

    Full Text Available Abstract Background Aflatoxin B1 (AFB1 is potent hepatotoxic and hepatocarcinogenic agent. In aflatoxicosis, oxidative stress is a common mechanism contributing to initiation and progression of hepatic damage. The aim of this work was to evaluate the hepatoprotective effect of cactus cladode extract (CCE on aflatoxin B1-induced liver damage in mice by measuring malondialdehyde (MDA level, the protein carbonyls generation and the heat shock proteins Hsp 70 and Hsp 27 expressions in liver. We also looked for an eventual protective effect against AFB1-induced genotoxicity as determined by chromosome aberrations test, SOS Chromotest and DNA fragmentation assay. We further evaluated the modulation of p53, bax and bcl2 protein expressions in liver. Methods Adult, healthy balbC (20-25 g male mice were pre-treated by intraperitonial administration of CCE (50 mg/Kg.b.w for 2 weeks. Control animals were treated 3 days a week for 4 weeks by intraperitonial administration of 250 μg/Kg.b.w AFB1. Animals treated by AFB1 and CCE were divided into two groups: the first group was administrated CCE 2 hours before each treatment with AFB1 3 days a week for 4 weeks. The second group was administrated without pre-treatment with CCE but this extract was administrated 24 hours after each treatment with AFB1 3 days a week for 4 weeks. Results Our results clearly showed that AFB1 induced significant alterations in oxidative stress markers. In addition, it has a genotoxic potential and it increased the expression of pro apoptotic proteins p53 and bax and decreased the expression of bcl2. The treatment of CCE before or after treatment with AFB1, showed (i a total reduction of AFB1 induced oxidative damage markers, (ii an anti-genotoxic effect resulting in an efficient prevention of chromosomal aberrations and DNA fragmentation compared to the group treated with AFB1 alone (iii restriction of the effect of AFB1 by differential modulation of the expression of p53 which

  12. New analytical techniques for mycotoxins in complex organic matrices. [Aflatoxins B1, B2, G1, and G2

    Energy Technology Data Exchange (ETDEWEB)

    Bicking, M.K.L.

    1982-07-01

    Air samples are collected for analysis from the Ames Solid Waste Recovery System. The high level of airborne fungi within the processing area is of concern due to the possible presence of toxic mycotoxins, and carcinogenic fungal metabolites. An analytical method has been developed to determine the concentration of aflatoxins B1, B2, G1, and G2 in the air of the plant which produces Refuse Derived Fuel (RDF). After extraction with methanol, some components in the matrix are precipitated by dissolving the sample in 30% acetonitrile/chloroform. An aliquot of this solution is injected onto a Styragel column where the sample components undergo simultaneous size exclusion and reverse phase partitioning. Additional studies have provided a more thorough understanding of solvent related non-exclusion effects on size exclusion gels. The Styragel column appears to have a useable lifetime of more than six months. After elution from Styragel, the sample is diverted to a second column containing Florisil which has been modified with oxalic acid and deactivated with water. Aflatoxins are eluted with 5% water/acetone. After removal of this solvent, the sample is dissolved in 150 ..mu..L of a spotting solvent and the entire sample applied to a thin layer chromatography (TLC) plate using a unique sample applicator developed here. The aflatoxins on the TLC plate are analyzed by laser fluorescence. A detection limit of 10 pg is possible for aflatoxin standards using a nitrogen laser as the excitation source. Sample concentrations are determined by comparing with an internal standard, a specially synthesized aflatoxin derivative. In two separate RDF samples, aflatoxin B1 was found at levels of 6.5 and 17.0 ppB. The analytical method has also proven useful in the analysis of contaminated corn and peanut meal samples. 42 figures, 8 tables.

  13. Quantifying Aflatoxin B1 in peanut oil using fabricating fluorescence probes based on upconversion nanoparticles

    Science.gov (United States)

    Sun, Cuicui; Li, Huanhuan; Koidis, Anastasios; Chen, Quansheng

    2016-08-01

    Rare earth doped upconversion nanoparticles convert near-infrared excitation light into visible emission light. Compared to organic fluorophores and semiconducting nanoparticles, upconversion nanoparticles (UCNPs) offer high photochemical stability, sharp emission bandwidths, and large anti-Stokes shifts. Along with the significant light penetration depth and the absence of autofluorescence in biological samples under infrared excitation, these UCNPs have attracted more and more attention on toxin detection and biological labelling. Herein, the fluorescence probe based on UCNPs was developed for quantifying Aflatoxin B1 (AFB1) in peanut oil. Based on a specific immunity format, the detection limit for AFB1 under optimal conditions was obtained as low as 0.2 ng·ml- 1, and in the effective detection range 0.2 to 100 ng·ml- 1, good relationship between fluorescence intensity and AFB1 concentration was achieved under the linear ratios up to 0.90. Moreover, to check the feasibility of these probes on AFB1 measurements in peanut oil, recovery tests have been carried out. A good accuracy rating (93.8%) was obtained in this study. Results showed that the nanoparticles can be successfully applied for sensing AFB1 in peanut oil.

  14. Thermal treatment of bentonite reduces aflatoxin b1 adsorption and affects stem cell death.

    Science.gov (United States)

    Nones, Janaína; Nones, Jader; Riella, Humberto Gracher; Poli, Anicleto; Trentin, Andrea Gonçalves; Kuhnen, Nivaldo Cabral

    2015-10-01

    Bentonites are clays that highly adsorb aflatoxin B1 (AFB1) and, therefore, protect human and animal cells from damage. We have recently demonstrated that bentonite protects the neural crest (NC) stem cells from the toxicity of AFB1. Its protective effects are due to the physico-chemical properties and chemical composition altered by heat treatment. The aim of this study is to prepare and characterize the natural and thermal treatments (125 to 1000 °C) of bentonite from Criciúma, Santa Catarina, Brazil and to investigate their effects in the AFB1 adsorption and in NC cell viability after challenging with AFB1. The displacement of water and mineralogical phases transformations were observed after the thermal treatments. Kaolinite disappeared at 500 °C and muscovite and montmorillonite at 1000 °C. Slight changes in morphology, chemical composition, and density of bentonite were observed. The adsorptive capacity of the bentonite particles progressively reduced with the increase in temperature. The observed alterations in the structure of bentonite suggest that the heat treatments influence its interlayer distance and also its adsorptive capacity. Therefore, bentonite, even after the thermal treatment (125 to 1000 °C), is able to increase the viability of NC stem cells previously treated with AFB1. Our results demonstrate the effectiveness of bentonite in preventing the toxic effects of AFB1.

  15. MicroRNA-24 Modulates Aflatoxin B1-Related Hepatocellular Carcinoma Prognosis and Tumorigenesis

    Directory of Open Access Journals (Sweden)

    Yi-Xiao Liu

    2014-01-01

    Full Text Available MicroRNA-24 (miR-24 may be involved in neoplastic process; however, the role of this microRNA in the hepatocellular carcinoma (HCC related to aflatoxin B1 (AFB1 has not been well elaborated. Here, we tested miR-24 expression in 207 pathology-diagnosed HCC cases from high AFB1 exposure areas and HCC cells. We found that miR-24 was upregulated in HCC tumor tissues relative to adjacent noncancerous tissue samples, and that the high expression of miR-24 was significantly correlated with larger tumor size, higher microvessel density, and tumor dedifferentiation. Additionally, this microRNA overexpression modified the recurrence-free survival (relative hazard ratio [HR], 4.75; 95% confidence interval [CI], 2.66–8.47 and overall survival (HR=3.58, 95% CI = 2.34–5.46 of HCC patients. Furthermore, we observed some evidence of joint effects between miR-24 and AFB1 exposure on HCC prognosis. Functionally, miR-24 overexpression progressed tumor cells proliferation, inhibited cell apoptosis, and developed the formation of AFB1-DNA adducts. These results indicate for the first time that miR-24 may modify AFB1-related HCC prognosis and tumorigenesis.

  16. Affinity improvement by fine tuning of single-chain variable fragment against aflatoxin B1.

    Science.gov (United States)

    Min, Won-Ki; Na, Kang-In; Yoon, Jung-Hyun; Heo, Yoon-Jee; Lee, Daesang; Kim, Sung-Gun; Seo, Jin-Ho

    2016-10-15

    Aflatoxin B1 (AFB1) produced in Aspergillus flavus is a major hepatocarcinogen found in foods and feed. For effective immunological detection of AFB1 at low concentrations, the development of high affinity antibody for AFB1 is required. Previously, an affinity-maturated single-chain variable fragment containing 6 mutations (scFv-M37) was isolated from an artificial mutagenic library, which showed a 9-fold higher affinity than its wild type scFv. In this study, the effect of the 6 mutated residues on the affinity improvement was characterized using surface plasmon resonance analysis, which identified a deleterious mutation (VH-A110T) located on a framework region of the scFv-M37. The back mutation of VH-A110T resulted in a 3.2-fold affinity improvement, which was attributed to decrease of dissociation rate constant (kd) in interaction between AFB1 and the back mutant scFv. The biophysical analyses using circular dichroism and gel filtration revealed that the back mutation of VH-A110T caused a subtle conformational change of the scFv toward tighter binding to AFB1. PMID:27173568

  17. Determination of Aflatoxin B1 Levels in Organic Spices and Herbs

    Directory of Open Access Journals (Sweden)

    Halil Tosun

    2013-01-01

    Full Text Available Organically produced spices and herbs were analyzed for determination of aflatoxin B1 (AFB1 by ELISA using immunoaffinity column. For this purpose 93 organic spices and 37 organic herbs were randomly selected from organic markets and organic shops in Turkey. AFB1 was detected in 58 organic spice and 32 organic herb samples. Among organic spice samples, the maximum value was detected in cinnamon sample (53 μg/kg. AFB1 was not detected in thyme samples. AFB1 levels of 41 organic spice samples were above the EU regulatory limit (5 μg/kg. Among organic herb samples the highest concentration of AFB1 (52.5 μg/kg was detected in a rosehip sample. AFB1 levels of 21 organic herb samples were above the regulatory limits of the European Union. These results showed that more stringent measures must be taken for the prevention of mold contamination in the production of organic spices and herbs.

  18. Aflatoxin B1 in sesame seeds and sesame products from the Greek market.

    Science.gov (United States)

    Kollia, Eleni; Tsourouflis, Kyriakos; Markaki, Panagiota

    2016-09-01

    Aflatoxin B1 (AFB1) is considered as the most potent liver carcinogen for humans. A method for determination in sesame seeds was developed. AFB1 was extracted by methanol-water, cleaned by immunoaffinity columns and determined by high-performance liquid chromatography with fluorescence detection. The recovery factor and the limit of detection (LOD) of AFB1 in sesame seeds were 111.5% and 0.02 ng g(-1), respectively. Thirty samples of sesame products were examined for the presence of AFB1. After analysis, 77.6% of samples were found to be contaminated. Eight samples exceeded the European Union (EU) limit (2 µg AFB1 kg(-1)). In 15 samples, AFB1 was below the EU limit. Seven samples remained below the LOD. The most contaminated (14.49 ng AFB1 g(-1)) sample was unpeeled packaged sesame seeds. In all samples, aflatoxigenic Aspergilli fungi as well as the risk for AFB1 presence in sesame seed was investigated.

  19. Cashew (Anacardium occidentale apple juice lowers mutagenicity of aflatoxin B1 in S. typhimurium TA102

    Directory of Open Access Journals (Sweden)

    Ana Amélia Melo Cavalcante

    2005-01-01

    Full Text Available Cashew (Anacardium occidentale is a medicinal plant native to Brazil and also yields a nutritious fruit juice. Its large pulpy pseudo-fruit, referred to as the cashew apple, contains high concentrations of vitamin C, carotenoids, phenolic compounds and minerals. Natural and processed cashew apple juice (CAJ/cajuina are amongst the most popular juices in Brazil, especially in the north-east. Both juices have antioxidant potential and suppress mutagenicity of hydrogen peroxide. In the present study we evaluated the inhibitory effects of CAJ/cajuina on Aflatoxin B1(AFB1-induced mutation, using the Salmonella/microsome assay with the experimental approaches of pre-, co- and post-treatments. Both CAJ/cajuina suppress AFB1-induced mutagenesis in strain TA102 when applied in co- and in post-treatment. Possible mechanisms for anti-mutagenicity in co-treatment are (a interaction with S9 enzymes, (b metabolization to non-mutagenic compounds of AFB1 or (c inactivation of S9 potential. Total suppression of AFB1 mutagenicity was observed in co-treatment with both CAJ and cajuina. Post-treatment anti-mutagenicity of both juices suggests a modulation of activity of error-prone DNA repair. CAJ/cajuina may be considered promising candidates for control of genotoxicity of AFB1 and may thus be considered as health foods with anti-carcinogenic potential. This promising characteristic warrants further evaluation with in vivo studies.

  20. Hepatic Transcriptome Responses of Domesticated and Wild Turkey Embryos to Aflatoxin B1

    Science.gov (United States)

    Monson, Melissa S.; Cardona, Carol J.; Coulombe, Roger A.; Reed, Kent M.

    2016-01-01

    The mycotoxin, aflatoxin B1 (AFB1) is a hepatotoxic, immunotoxic, and mutagenic contaminant of food and animal feeds. In poultry, AFB1 can be maternally transferred to embryonated eggs, affecting development, viability and performance after hatch. Domesticated turkeys (Meleagris gallopavo) are especially sensitive to aflatoxicosis, while Eastern wild turkeys (M. g. silvestris) are likely more resistant. In ovo exposure provided a controlled AFB1 challenge and comparison of domesticated and wild turkeys. Gene expression responses to AFB1 in the embryonic hepatic transcriptome were examined using RNA-sequencing (RNA-seq). Eggs were injected with AFB1 (1 μg) or sham control and dissected for liver tissue after 1 day or 5 days of exposure. Libraries from domesticated turkey (n = 24) and wild turkey (n = 15) produced 89.2 Gb of sequence. Approximately 670 M reads were mapped to a turkey gene set. Differential expression analysis identified 1535 significant genes with |log2 fold change| ≥ 1.0 in at least one pair-wise comparison. AFB1 effects were dependent on exposure time and turkey type, occurred more rapidly in domesticated turkeys, and led to notable up-regulation in cell cycle regulators, NRF2-mediated response genes and coagulation factors. Further investigation of NRF2-response genes may identify targets to improve poultry resistance. PMID:26751476

  1. Hepatic Transcriptome Responses of Domesticated and Wild Turkey Embryos to Aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Melissa S. Monson

    2016-01-01

    Full Text Available The mycotoxin, aflatoxin B1 (AFB1 is a hepatotoxic, immunotoxic, and mutagenic contaminant of food and animal feeds. In poultry, AFB1 can be maternally transferred to embryonated eggs, affecting development, viability and performance after hatch. Domesticated turkeys (Meleagris gallopavo are especially sensitive to aflatoxicosis, while Eastern wild turkeys (M. g. silvestris are likely more resistant. In ovo exposure provided a controlled AFB1 challenge and comparison of domesticated and wild turkeys. Gene expression responses to AFB1 in the embryonic hepatic transcriptome were examined using RNA-sequencing (RNA-seq. Eggs were injected with AFB1 (1 μg or sham control and dissected for liver tissue after 1 day or 5 days of exposure. Libraries from domesticated turkey (n = 24 and wild turkey (n = 15 produced 89.2 Gb of sequence. Approximately 670 M reads were mapped to a turkey gene set. Differential expression analysis identified 1535 significant genes with |log2 fold change| ≥ 1.0 in at least one pair-wise comparison. AFB1 effects were dependent on exposure time and turkey type, occurred more rapidly in domesticated turkeys, and led to notable up-regulation in cell cycle regulators, NRF2-mediated response genes and coagulation factors. Further investigation of NRF2-response genes may identify targets to improve poultry resistance.

  2. Response of the hepatic transcriptome to aflatoxin B1 in domestic turkey (Meleagris gallopavo).

    Science.gov (United States)

    Monson, Melissa S; Settlage, Robert E; McMahon, Kevin W; Mendoza, Kristelle M; Rawal, Sumit; El-Nezami, Hani S; Coulombe, Roger A; Reed, Kent M

    2014-01-01

    Dietary exposure to aflatoxin B1 (AFB1) is detrimental to avian health and leads to major economic losses for the poultry industry. AFB1 is especially hepatotoxic in domestic turkeys (Meleagris gallopavo), since these birds are unable to detoxify AFB1 by glutathione-conjugation. The impacts of AFB1 on the turkey hepatic transcriptome and the potential protection from pretreatment with a Lactobacillus-based probiotic mixture were investigated through RNA-sequencing. Animals were divided into four treatment groups and RNA was subsequently recovered from liver samples. Four pooled RNA-seq libraries were sequenced to produce over 322 M reads totaling 13.8 Gb of sequence. Approximately 170,000 predicted transcripts were de novo assembled, of which 803 had significant differential expression in at least one pair-wise comparison between treatment groups. Functional analysis linked many of the transcripts significantly affected by AFB1 exposure to cancer, apoptosis, the cell cycle or lipid regulation. Most notable were transcripts from the genes encoding E3 ubiquitin-protein ligase Mdm2, osteopontin, S-adenosylmethionine synthase isoform type-2, and lipoprotein lipase. Expression was modulated by the probiotics, but treatment did not completely mitigate the effects of AFB1. Genes identified through transcriptome analysis provide candidates for further study of AFB1 toxicity and targets for efforts to improve the health of domestic turkeys exposed to AFB1.

  3. Response of the hepatic transcriptome to aflatoxin B1 in domestic turkey (Meleagris gallopavo.

    Directory of Open Access Journals (Sweden)

    Melissa S Monson

    Full Text Available Dietary exposure to aflatoxin B1 (AFB1 is detrimental to avian health and leads to major economic losses for the poultry industry. AFB1 is especially hepatotoxic in domestic turkeys (Meleagris gallopavo, since these birds are unable to detoxify AFB1 by glutathione-conjugation. The impacts of AFB1 on the turkey hepatic transcriptome and the potential protection from pretreatment with a Lactobacillus-based probiotic mixture were investigated through RNA-sequencing. Animals were divided into four treatment groups and RNA was subsequently recovered from liver samples. Four pooled RNA-seq libraries were sequenced to produce over 322 M reads totaling 13.8 Gb of sequence. Approximately 170,000 predicted transcripts were de novo assembled, of which 803 had significant differential expression in at least one pair-wise comparison between treatment groups. Functional analysis linked many of the transcripts significantly affected by AFB1 exposure to cancer, apoptosis, the cell cycle or lipid regulation. Most notable were transcripts from the genes encoding E3 ubiquitin-protein ligase Mdm2, osteopontin, S-adenosylmethionine synthase isoform type-2, and lipoprotein lipase. Expression was modulated by the probiotics, but treatment did not completely mitigate the effects of AFB1. Genes identified through transcriptome analysis provide candidates for further study of AFB1 toxicity and targets for efforts to improve the health of domestic turkeys exposed to AFB1.

  4. Effects of Nutrients in Substrates of Different Grains on Aflatoxin B1 Production by Aspergillus flavus

    Science.gov (United States)

    Liu, Jie; Sun, Lvhui; Zhang, Niya; Zhang, Jiacai; Guo, Jiao; Li, Chong; Rajput, Shahid Ali; Qi, Desheng

    2016-01-01

    The current study was to better understand the potential factors affecting aflatoxin B1 (AFB1) accumulation varies between different grains. The nutrient composition and contents of defatted substrates were determined; additionally, according to the nutrient content of the substrates, the effects of starch, soluble sugars, amino acids, and trace elements on AFB1 production and mycelial growth in Czapek-Dox medium were examined. These results verified that removal of lipids from ground substrates significantly reduced the substrate's potential for AFB1 production by Aspergillus flavus. Maltose, glucose, sucrose, arginine, glutamic acid, aspartic acid, and zinc significantly induced AFB1 production up to 1.7- to 26.6-fold. And stachyose more significantly promoted A. flavus growth than the other nutrients. Thus, this study demonstrated that, combined with the nutrients content of grains, in addition to lipids, sucrose, stachyose, glutamic acid, and zinc might play key roles in various grains that are differentially infected by A. flavus. Particularly, two new nutrients (arginine and stachyose) of the grains we found significantly stimulate AFB1 production and A. flavus growth, respectively. The results provide new concepts for antifungal methods to protect food and animal feed from AFB1 contamination. PMID:27294129

  5. Bioactivation and Regioselectivity of Pig Cytochrome P450 3A29 towards Aflatoxin B1

    Science.gov (United States)

    Wu, Jun; Chen, Ruohong; Zhang, Caihui; Li, Kangbai; Xu, Weiying; Wang, Lijuan; Chen, Qingmei; Mu, Peiqiang; Jiang, Jun; Wen, Jikai; Deng, Yiqun

    2016-01-01

    Due to unavoidable contaminations in feedstuff, pigs are easily exposed to aflatoxin B1 (AFB1) and suffer from poisoning, thus the poisoned products potentially affect human health. Heretofore, the metabolic process of AFB1 in pigs remains to be clarified, especially the principal cytochrome P450 oxidases responsible for its activation. In this study, we cloned CYP3A29 from pig liver and expressed it in Escherichia coli, and its activity has been confirmed with the typical P450 CO-reduced spectral characteristic and nifedipine-oxidizing activity. The reconstituted membrane incubation proved that the recombinant CYP3A29 was able to oxidize AFB1 to form AFB1-exo-8,9-epoxide in vitro. The structural basis for the regioselective epoxidation of AFB1 by CYP3A29 was further addressed. The T309A mutation significantly decreased the production of AFBO, whereas F304A exhibited an enhanced activation towards AFB1. In agreement with the mutagenesis study, the molecular docking simulation suggested that Thr309 played a significant role in stabilization of AFB1 binding in the active center through a hydrogen bond. In addition, the bulk phenyl group of Phe304 potentially imposed steric hindrance on the binding of AFB1. Our study demonstrates the bioactivation of pig CYP3A29 towards AFB1 in vitro, and provides the insight for understanding regioselectivity of CYP3A29 to AFB1. PMID:27626447

  6. Aflatoxin B1: Toxicity, bioactivation and detoxification in the polyphagous caterpillar Trichoplusia ni

    Institute of Scientific and Technical Information of China (English)

    Ren Sen Zeng; Zhimou Wen; Guodong Niu; May R.Berenbaum

    2013-01-01

    Trichoplusia ni caterpillars are polyphagous foliage-feeders and rarely likely to encounter aflatoxin B1 (AFB1),a mycotoxin produced by Aspergillus flavus and A.parasiticus,in their host plants.To determine how T.ni copes with AFB1,we evaluated the toxicity ofAFB1 to T.ni caterpillars at different developmental stages and found that AFB1 tolerance significantly increases with larval development.Diet incorporation of AFB1 at 1μg/g completely inhibited larval growth and pupation of newly hatched larvae,but 3μg/g AFB1 did not have apparent toxic effects on larval growth and pupation of caterpillars that first consume this compound 10 days after hatching.Piperonyl butoxide,a general inhibitor of cytochrome P450 monooxygenases (P450s),reduced the toxicity of AFB1,suggesting that AFB1 is bioactivated in T.ni and this bioactivation is mediated by P450s.Some plant allelochemicals,including flavonoids such as flavones,furanocoumarins such as xanthotoxin and imperatorin,and furanochromones such as visnagin,that induce P450s in other lepidopteran larvae ameliorated AFB1 toxicity,suggesting that P450s are also involved in AFB1 detoxification in T.ni.

  7. Genotoxic evaluation of ammonium inactivated aflatoxin B1 in mice fed with contaminated corn.

    Science.gov (United States)

    Márquez-Márquez, R; Madrigal-Bujaidar, E; Tejada de Hernández, I

    1993-03-01

    Aflatoxin B1 (AFB1) is a major contaminant in different agricultural products including maize. In an attempt to reduce this problem and the hazards to human health, an AFB1 inactivating system with ammonia has been developed. In this work we evaluated the efficiency of the system in mice using micronucleus (MN) and sister chromatid exchange (SCE) analysis. Four groups of animals were fed during 8 weeks with a special diet mainly composed of maize: (1) uncontaminated; (2) uncontaminated/inactivated; (3) contaminated/inactivated; and (4) contaminated. We evaluated MN at weekly intervals in peripheral blood, and in weeks 4 and 8 SCE frequencies were quantified in bone marrow cells. The results showed that animals fed with AFB1 contaminated/inactivated maize had a 45% lower level of induced cytogenetic damage than those animals fed with AFB1 contaminated, but not inactivated maize. A residual amount of AFB1 after the inactivating treatment and the reconversion back to AFB1 in the organism may account for the remaining increased levels of SCE and MN.

  8. Degradation of aflatoxin B1 during the fermentation of alcoholic beverages.

    Science.gov (United States)

    Inoue, Tomonori; Nagatomi, Yasushi; Uyama, Atsuo; Mochizuki, Naoki

    2013-06-28

    Aflatoxin B1 (AFB1) is a contaminant of grain and fruit and has one of the highest levels of carcinogenicity of any natural toxin. AFB1 and the fungi that produce it can also contaminate the raw materials used for beer and wine manufacture, such as corn and grapes. Therefore, brewers must ensure strict monitoring to reduce the risk of contamination. In this study, the fate of AFB1 during the fermentation process was investigated using laboratory-scale bottom and top beer fermentation and wine fermentation. During fermentation, cool wort beer samples and wine must samples were artificially spiked with AFB1 and the levels of AFB1 remaining after fermentation were analyzed. AFB1 levels were unchanged during both types of fermentation used for beer but were reduced to 30% of their initial concentration in wine. Differential analysis of the spiked and unspiked wine samples showed that the degradation compound was AFB2a, a hydrated derivative of AFB1. Thus, the results showed that the risk of AFB1 carryover was still present for both types of beer fermentation but was reduced in the case of wine fermentation because of hydration.

  9. Determination of aflatoxin B1 levels in organic spices and herbs.

    Science.gov (United States)

    Tosun, Halil; Arslan, Recep

    2013-01-01

    Organically produced spices and herbs were analyzed for determination of aflatoxin B1 (AFB1) by ELISA using immunoaffinity column. For this purpose 93 organic spices and 37 organic herbs were randomly selected from organic markets and organic shops in Turkey. AFB1 was detected in 58 organic spice and 32 organic herb samples. Among organic spice samples, the maximum value was detected in cinnamon sample (53 μg/kg). AFB1 was not detected in thyme samples. AFB1 levels of 41 organic spice samples were above the EU regulatory limit (5 μg/kg). Among organic herb samples the highest concentration of AFB1 (52.5 μg/kg) was detected in a rosehip sample. AFB1 levels of 21 organic herb samples were above the regulatory limits of the European Union. These results showed that more stringent measures must be taken for the prevention of mold contamination in the production of organic spices and herbs.

  10. Effect of aflatoxin-B1 doses simulating natural food contamination reproductive steroid hormones in rats

    International Nuclear Information System (INIS)

    Aflatoxin B1 (AFB1) is the most toxic metabolite synthesized by aspergillus flavus. The mycotoxins was found to be endemic contaminant in underdeveloped countries and in egypt was documented as a pollutant of a wide variety of products for human and animal nutrition. Carcinogenic and mutagenic effects of AFB1 has been investigated extensively, while very scare information is available about other possible endocrine effects of the toxin which might precedes carcinogenic effects. This study was performed to investigate the effects of in vivo administration of AFB1 via intraperitoneal injection (I.P) in adult male rats to show its effects on rat reproductive function and to illucidate the effects of acute, chronic and sub toxic (endimomemitic) AFB1 doses on male rat steroid function. Intraperitoneal injection (I.P) of AFB 1 doses in adult male rats revealed that AFB1 caused significant decrease in serum testosterone and cortisol (early), while a significant increase was observed in progesterone (P4) and Estrodial (E2) (late)

  11. Application of lactic acid bacteria in removing heavy metals and aflatoxin B1 from contaminated water.

    Science.gov (United States)

    Elsanhoty, Rafaat M; Al-Turki, I A; Ramadan, Mohamed Fawzy

    2016-01-01

    In this study selected lactic acid bacteria (LAB, Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus plantrium and Streptococcus thermophiles) and probiotic bacteria (Bifidobacterium angulatum) were tested for their ability in removing heavy metals (HM) including cadmium (Cd), lead (Pb) and arsenic (As) as well as aflatoxin B1 (AFB1) from contaminated water. The biosorption parameters (pH, bacterial concentration, contact time and temperature) of removal using individual as well as mixed LAB and probiotic bacteria were studied. Removal of HM and AFB1 depended on the strain, wherein the process was strongly pH-dependent with high removal ability at a pH close to neutral. The increase in bacterial concentration enhanced the removal of Cd, Pb and As. Also, increasing of contact time and temperature increased the ability of LAB to remove HM. The effect of contact time on Cd removal was slightly different when freshly cultured cells were used. The removal of Cd, Pb and As decreased with the increase in the initial metal concentration. The most effective HM removers were Lactobacillus acidophilus and Bifidobacterium angulatum. The system was found to be adequate for concentrations of HM under investigation. At the end of the operation, the concentration of HM reached the level allowed by the World Health Organization regulations. PMID:27508367

  12. An aptamer-based dipstick assay for the rapid and simple detection of aflatoxin B1.

    Science.gov (United States)

    Shim, Won-Bo; Kim, Min Jin; Mun, Hyoyoung; Kim, Min-Gon

    2014-12-15

    A rapid and simple dipstick assay based on an aptamer has been developed for the determination of aflatoxin B1 (AFB1). The dipstick assay format was based on a competitive reaction of the biotin-modified aptamer specific to AFB1 between target and cy5-modified DNA probes. Streptavidin and anti-cy5 antibody as capture reagents were immobilized at test and control lines on a membrane of the dipstick assay. After optimization, the limit of detection for the dipstick assay was 0.1 ng/ml AFB1 in buffer. The method was confirmed to be specific to AFB1, and the entire process of the assay can be completed within 30 min. Aqueous methanol (20%) provided a good extraction efficiency, and the matrix influence from corn extracts was successfully reduced through 2-fold dilution. The results of AFB1 analysis for corn samples spiked with known concentration of AFB1 by the dipstick assay and ELISA showed good agreement. The cut-off value of the dipstick assay for corn samples was 0.3 ng/g AFB1. Therefore, the dipstick assay is first reported and considered as a rapid, simple, on-site and inexpensive screening tool for AFB1 determination in grains as well as a corn. PMID:25032679

  13. [Contamination level of aflatoxin B1 in lotus seeds rapid screening by indirect competitive ELISA method].

    Science.gov (United States)

    Chu, Xian-feng; Dou, Xiao-wen; Kong, Wei-jun; Yang, Mei-hua; Zhao, Chong; Zhao, Ming; Ouyang, Zhen

    2015-02-01

    A simple and cost-effective indirect competitive enzyme-linked immune sorbent assay (ic-ELISA) was developed to rapidly screen the content of aflatoxin B1 (AFB1) in lotus seeds, and the results were confirmed by ultra-fast liquid chromatography-tandem mass spectrometry( UFLC-MS/MS). Matrix-matched calibration expressed a good linearity ranging from 0. 171 to 7. 25 µg · L(-1) for AFB, with R2 > 0.978. The medium inhibitory concentration( IC50 ) for AFB1 was 1.29 µg · L(-1), the recovery for AFB1 was 74.73% to 126.9% with RSD lotus seeds samples and the results indicated that the contents of AFB, in samples 1-15 were in the range of 1. 19- 115. 3 µg · kg(-1) and in 40% of the samples exceeded the legal limit(5 µg · kg(-1)), while the contamination rate of AFB, in samples 16-20 was 40%. Pearson correlation coefficient(r) reached 0.997 for AFB1 content in the samples detected by ic-ELSIA and UFLC-MS/MS methods. The results proved that the developed ic-ELISA method is simple, sensitive and reliable, and can be used for rapid and high-throughput screening of AFB1 in lotus seeds

  14. In-vitro cytotoxicity of aflatoxin B1 to broiler lymphocytes of broiler chickens

    Directory of Open Access Journals (Sweden)

    CEP Zimmermann

    2014-09-01

    Full Text Available The aim of the present work was to study the in-vitro cytotoxic effects of different concentrations of aflatoxin B1 (AFB1 on broiler lymphocytes. Lymphocyte-rich mononuclear cells were separated by Ficoll-Histopaque density and cultured in 96-wellplates containing the evaluated AFB1 concentrations in 5% CO2 atmosphere at 39°C. Thereafter, MTT, PicoGreen, and reactive oxygen species assays were performed. Cell viability decreased in the presence of 10 µg/mL AFB1 at 48 h (p < 0.05 and of 10 and 20 µg/mL AFB1 at 72 h (p < 0.01 and p < 0.001, respectively when compared to the control (0 µg/mL. However, a dose-dependent increase in the cell-free DNA at 24 h was observed at 1, 10 and 20 µg/mL (p < 0.001. ROS formation significantly increased at 24 h at all concentrations (p < 0.001. The in-vitro results demonstrate that AFB1 is cytotoxic and causes biomolecular oxidative damage in broiler lymphocytes.

  15. Aflatoxin B1 up-regulates insulin receptor substrate 2 and stimulates hepatoma cell migration.

    Directory of Open Access Journals (Sweden)

    Yanli Ma

    Full Text Available Aflatoxin B1 (AFB1 is a potent carcinogen that can induce hepatocellular carcinoma. AFB1-8,9-exo-epoxide, one of AFB1 metabolites, acts as a mutagen to react with DNA and induce gene mutations, including the tumor suppressor p53. In addition, AFB1 reportedly stimulates IGF receptor activation. Aberrant activation of IGF-I receptor (IGF-IR signaling is tightly associated with various types of human tumors. In the current study, we investigated the effects of AFB1 on key elements in IGF-IR signaling pathway, and the effects of AFB1 on hepatoma cell migration. The results demonstrated that AFB1 induced IGF-IR, Akt, and Erk1/2 phosphorylation in hepatoma cell lines HepG2 and SMMC-7721, and an immortalized human liver cell line Chang liver. AFB1 also down-regulated insulin receptor substrate (IRS 1 but paradoxically up-regulated IRS2 through preventing proteasomal degradation. Treatment of hepatoma cells and Chang liver cells with IGF-IR inhibitor abrogated AFB1-induced Akt and Erk1/2 phosphorylation. In addition, IRS2 knockdown suppressed AFB1-induced Akt and Erk1/2 phosphorylation. Finally, AFB1 stimulated hepatoma cell migration. IGF-IR inhibitor or IRS2 knockdown suppressed AFB1-induced hepatoma cell migration. These data demonstrate that AFB1 stimulates hepatoma cell migration through IGF-IR/IRS2 axis.

  16. Effects of Nutrients in Substrates of Different Grains on Aflatoxin B1 Production by Aspergillus flavus.

    Science.gov (United States)

    Liu, Jie; Sun, Lvhui; Zhang, Niya; Zhang, Jiacai; Guo, Jiao; Li, Chong; Rajput, Shahid Ali; Qi, Desheng

    2016-01-01

    The current study was to better understand the potential factors affecting aflatoxin B1 (AFB1) accumulation varies between different grains. The nutrient composition and contents of defatted substrates were determined; additionally, according to the nutrient content of the substrates, the effects of starch, soluble sugars, amino acids, and trace elements on AFB1 production and mycelial growth in Czapek-Dox medium were examined. These results verified that removal of lipids from ground substrates significantly reduced the substrate's potential for AFB1 production by Aspergillus flavus. Maltose, glucose, sucrose, arginine, glutamic acid, aspartic acid, and zinc significantly induced AFB1 production up to 1.7- to 26.6-fold. And stachyose more significantly promoted A. flavus growth than the other nutrients. Thus, this study demonstrated that, combined with the nutrients content of grains, in addition to lipids, sucrose, stachyose, glutamic acid, and zinc might play key roles in various grains that are differentially infected by A. flavus. Particularly, two new nutrients (arginine and stachyose) of the grains we found significantly stimulate AFB1 production and A. flavus growth, respectively. The results provide new concepts for antifungal methods to protect food and animal feed from AFB1 contamination. PMID:27294129

  17. Synergistic effect of black tea and curcumin in improving the hepatotoxicity induced by aflatoxin B1 in rats.

    Science.gov (United States)

    Alm-Eldeen, Abeer A; Mona, Mohamed H; Shati, Ali A; El-Mekkawy, Haitham I

    2015-12-01

    Aflatoxin B1 (AFB1) is a toxic compound commonly found as a contaminant in human food. It is carcinogenic due its potential in inducing the oxidative stress and distortion of the most antioxidant enzymes. Since black tea possesses strong antioxidant activity, it protects cells and tissues against oxidative stress. Curcumin (CMN), a naturally occurring agent, has a combination of biological and pharmacological properties that include antioxidant activity. Therefore, the present study was carried out to investigate the possible role of separate and mixed supplementation of black tea extract and CMN in the hepatotoxicity induced by AFB1 in rats. A total of 48: adult male Sprague Dawley rats were randomly divided into eight groups with six rats in each group. Group 1 (normal control) includes rats that received no treatment. Groups 2, 3, and 4 (positive control) include rats that received olive oil, black tea extract, and CMN, respectively. Group 5 includes rats that received AFB1 at a dose of 750 μg/kg body weight (b.w.) dissolved in olive oil. Groups 6, 7, and 8 include rats that received AFB1 along with 2% black tea extract, CMN at a dose of 200 mg/kg b.w., and both black tea extract and CMN at the same previous doses, respectively. After 90 days, biochemical and histopathological examination was carried out for the blood samples and liver tissues. A significant decrease in the antioxidant enzymes and a significant increase in the lipid peroxidation and hydrogen peroxide in the rats treated with AFB1 were observed. Moreover, there were dramatic changes in the liver function biomarkers, lipid profile, and liver architecture. Supplementation of black tea extract or CMN showed an efficient role in repairing the distortion of the biochemical and histological changes induced by AFB1 in liver. This improvement was more pronounced when both CMN and black tea were used together.

  18. Modification of aflatoxin B1 and ochratoxin A toxicokinetics in rats administered a yeast cell wall preparation

    OpenAIRE

    Firmin, Stéphane; Gandia, Peggy; Morgavi, Diego Pablo; Houin, Georges; Jouany, JP; Bertin, Gérard; Boudra, Hamid

    2010-01-01

    Abstract The cell wall of Saccharomyces cerevisiae can bind mycotoxins in vitro but there is scarce information on whether this property decreases the absorption of mycotoxins in vivo. The effect of a yeast cell wall preparation (YCW) on toxicokinetics and balance excretion (urine and faeces) of aflatoxin B1 (AFB1) and ochratoxin A (OTA) was tested in rats after oral administration of each toxin. The 3H-labelled mycotoxins were used at low doses. Co-administration of YCW with AF...

  19. Cassia senna inhibits mutagenic activities of benzo[a]-pyrene, aflatoxin B1, shamma and methyl methanesulfonate.

    Science.gov (United States)

    al-Dakan, A A; al-Tuffail, M; Hannan, M A

    1995-10-01

    Ethanol extract of Senokot tablets (Cassia senna concentrate used as vegetable laxative), was found to be non-mutagenic while it inhibited the mutagenicity of benzo[a]pyrene, shamma, aflatoxin B1 and methyl methanesulfonate in the Ames histidine reversion assay using the Salmonella typhimurium tester strain TA98. While the Senokot extract completely inhibited the mutagenicity of promutagens (i.e. metabolic activation dependent) like benzo[a]pyrene and shamma, it reduced the mutagenic activity of the direct acting mutagen methyl methanesulfonate by only 58%. The mutagen aflatoxin B1 showed a 25-fold increase in the number of histidine revertants per plate at low concentrations (1.0-4.0 micrograms/plate) in the presence of metabolic activation system while at high concentrations (10.0-30.0 micrograms/plate) it proved to be weakly mutagenic (with a 5-fold increase in the number of histidine revertants/plate) without metabolic activation. The Senokot extract completely inhibited the mutagenic effect of low concentrations of aflatoxin B1 in the presence of metabolic activation but not that resulting from higher concentrations without metabolic activation. The results obtained with benzo[a]pyrene, shamma and aflatoxin B1 indicated that the antimutagenic effects of Senokot extract could be largely due to an interaction with the metabolic process involved in the activation of procarcinogens. However, the results obtained with methyl methanesulfonate suggested that factors in Senokot may also interact with direct mutagens to produce some antimutagenic effects. An ethanol extract of crude senna leaves found to be weakly mutagenic also inhibited (though less than Senokot) the mutagenic effect of benzo[a]pyrene suggesting that the antimutagenic principle is present in the complex plant material itself.

  20. Synthesis and Characterization of Different Crystalline Calcium Silicate Hydrate: Application for the Removal of Aflatoxin B1 from Aqueous Solution

    OpenAIRE

    Lu Zeng; Ligang Yang; Shuping Wang; Kai Yang

    2014-01-01

    Different crystalline calcium silicate hydrates (CSH) were synthesized under specific hydrothermal conditions and several methods were used to analyze samples. Amorphous calcium silicate hydrates (ACSH) mainly consists of disordered calcium silicate hydrate gel (C-S-H gel) and crystalline calcium silicate hydrates (CCSH) consists of crystallized tobermorite. The adsorption of carcinogenic aflatoxin B1 (AFB1) onto ACSH and CCSH was investigated. The adsorption kinetics was studied using pseudo...

  1. Immunoaffinity column cleanup with liquid chromatography for determination of aflatoxin B1 in corn samples: interlaboratory study.

    Science.gov (United States)

    Brera, Carlo; Debegnach, Francesca; Minardi, Valentina; Pannunzi, Elena; De Santis, Barbara; Miraglia, Marina

    2007-01-01

    An interlaboratory study was conducted to evaluate the effectiveness of an immunoaffinity column cleanup liquid chromatography (LC) method for the determination of aflatoxin B1 levels in corn samples, enforced by European Union legislation. A test portion was extracted with methanol-water (80 + 20); the extract was filtered, diluted with phosphate-buffered saline solution, filtered on a microfiber glass filter, and applied to an immunoaffinity column. The column was washed with deionized water to remove interfering compounds, and the purified aflatoxin B1 was eluted with methanol. Aflatoxin B1 was separated and determined by reversed-phase LC with fluorescence detection after either pre- or postcolumn derivatization. Precolumn derivatization was achieved by generating the trifluoroacetic acid derivative, used by 8 laboratories. The postcolumn derivatization was achieved either with pyridinium hydrobromide perbromide, used by 16 laboratories, or with an electrochemical cell by the addition of bromide to the mobile phase, used by 5 laboratories. The derivatization techniques used were not significantly different when compared by the Student's t-test; the method was statistically evaluated for all the laboratories. Five corn sample materials, both spiked and naturally contaminated, were sent to 29 laboratories (22 Italian and 7 European). Test portions were spiked with aflatoxin B1 at levels of 2.00 and 5.00 ng/g. The mean values for recovery were 82% for the low level and 84% for the high contamination level. Based on results for spiked samples (blind pairs at 2 levels) as well as naturally contaminated samples (blind pairs at 3 levels), the values for relative standard deviation for repeatability (RSDr) ranged from 9.9 to 28.7%. The values for relative standard deviation for reproducibility (RSDR) ranged from 18.6 to 36.8%. The method demonstrated acceptable within- and between-laboratory precision for this matrix, as evidenced by the HorRat values. PMID:17580628

  2. Cytoprotective effect of palm kernel cake phenolics against aflatoxin B1-induced cell damage and its underlying mechanism of action

    OpenAIRE

    Oskoueian, Ehsan; Abdullah, Norhani; Zulkifli, Idrus; Ebrahimi, Mahdi; Karimi, Ehsan; Goh, Yong Meng; Oskoueian, Armin; Shakeri, Majid

    2015-01-01

    Background Palm kernel cake (PKC), a by-product of the palm oil industry is abundantly available in many tropical and subtropical countries. The product is known to contain high levels of phenolic compounds that may impede the deleterious effects of fungal mycotoxins. This study focused on the evaluation of PKC phenolics as a potential cytoprotective agent towards aflatoxin B1 (AFB1)-induced cell damage. Methods The phenolic compounds of PKC were obtained by solvent extraction and the product...

  3. Immunochromatographic Assay for Ultrasensitive Detection of Aflatoxin B1 in Maize by Highly Luminescent Quantum Dot Beads

    OpenAIRE

    Ren, Meiling; Xu, Hengyi; Huang, Xiaolin; Kuang, Min; Xiong, Yonghua; Xu, Hong; Xu, Yang; Chen, Hongyu; Wang, Andrew

    2014-01-01

    Highly luminescent quantum dot beads (QBs) were synthesized by encapsulating CdSe/ZnS and used for the first time as immunochromatographic assay (ICA) signal amplification probe for ultrasensitive detection of aflatoxin B1 (AFB1) in maize. The challenges to using high brightness QBs as probes for ICA are smooth flow of QBs and nonspecific binding on nitrocellulose (NC) membrane, which are overcome by unique polymer encapsulation of quantum dots (QDs) and surface blocking method. Under optimal...

  4. Phytic Acid Exposure Alters AflatoxinB1-Induced Reproductive and Oxidative Toxicity in Albino Rats (Rattus norvegicus)

    OpenAIRE

    Abu El-Saad, Abdelaziz S.; Mahmoud, Hamada M.

    2009-01-01

    The increased use of feed in Egypt's aquaculture and animal industries raises concerns about the possible presence of mycotoxins in feedstuffs. The use of alternative medicine, such as botanicals and nutritional supplements, has become popular with inflammatory cases. The present study aimed to testify the role played by phytic acid (IP6) in enhancing the reproductive and oxidative toxicity induced in aflatoxinB1 (AFB1) treated white male albino rats (Rattus norvegicus) throughout treatment a...

  5. Mechanismen der Aktivierung und Detoxifizierung von Aflatoxin B 1 in verschiedenen Leberzelltypen von Ratten, Mäusen und Waldmurmeltieren

    OpenAIRE

    Gemechu, Mekonnen

    2000-01-01

    Zusammenfassung: Die Applikation des Mykotoxins Aflatoxin B1 (AFB1) führt in der Ratte zu Lebertumoren hepatozellulären Ursprungs, während bisher keine transformierende Wirkung dieses Mykotoxins auf Kupffer- und Endothelzellen (Nichtparenchymzellen, NPC) nachgewiesen werden konnte. Diese Resistenzmechanismen der NPC gegenüber AFB1 wurden im ersten Teil dieser Arbeit untersucht. AFB1 ist per se inaktiv, wird jedoch durch Verstoffwechselung in den chemisch reaktiven, an DNA bindenden Metabolit...

  6. Assessment of genotoxicity of aflatoxin M1 and B1 contaminated milks after in vitro human digestion

    OpenAIRE

    Ladeira, Carina; Becker-Algeri, Tania Aparecida; Pimenta, Andreia Isabel; Eliana BADIALE-FURLONG

    2016-01-01

    Introduction - Milk is considered a complete food from the nutritional point of view. Milk can be exposed to various types of contamination, such as mycotoxins. These metabolites are naturally occurring toxic compounds produced by fungi. Several studies on milk samples have reported the presence of aflatoxin B1 (AFB1) and M1 (AFM1), due to the high incidence in samples intended for human consumption, carcinogenicity proven AFB1 and resistance of the contaminants to the process of digestion, m...

  7. Biotransformation of aflatoxin B1 and aflatoxin G1 in peanut meal by anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus.

    Science.gov (United States)

    Chen, Yujie; Kong, Qing; Chi, Chen; Shan, Shihua; Guan, Bin

    2015-10-15

    The purpose of this study was to explore the ability of anaerobic solid fermentation of Streptococcus thermophilus and Lactobacillus delbrueckii subsp. bulgaricus to biotransform aflatoxins in peanut meal. The pH of the peanut meal was adjusted above 10, and then heated for 10 min at 100 °C, 115 °C and 121 °C. The S. thermophilus and L. delbrueckii subsp. bulgaricus were precultured together in MRS broth for 48 h at 37 °C. The heated peanut meal was mixed with precultured MRS broth containing 7.0×10(8) CFU/mL of S. thermophilus and 3.0×10(3) CFU/mL of L. delbrueckii subsp. bulgaricus with the ratio of 1 to 1 (weight to volume) and incubated in anaerobic jars at 37 °C for 3 days. The aflatoxin content in the peanut meal samples was determined by HPLC. The results showed that the peanut meal contained mainly aflatoxin B1 (AFB1) (10.5±0.64 μg/kg) and aflatoxin G1 (AFG1) (18.7±0.55 μg/kg). When heat treatment was combined with anaerobic solid fermentation, the biotransformation rate of aflatoxins in peanut meal could attain 100%. The cytotoxicity of fermented peanut meal to L929 mouse connective tissue fibroblast cells was determined by MTT assay and no significant toxicity was observed in the fermented peanut meal. Furthermore, heat treatment and anaerobic solid fermentation did not change the amino acid concentrations and profile in peanut meal.

  8. Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis

    Science.gov (United States)

    Wang, Du; Zhang, Zhaowei; Li, Peiwu; Zhang, Qi; Zhang, Wen

    2016-01-01

    Rapid and quantitative sensing of aflatoxin B1 with high sensitivity and specificity has drawn increased attention of studies investigating soybean sauce. A sensitive and rapid quantitative immunochromatographic sensing method was developed for the detection of aflatoxin B1 based on time-resolved fluorescence. It combines the advantages of time-resolved fluorescent sensing and immunochromatography. The dynamic range of a competitive and portable immunoassay was 0.3–10.0 µg·kg−1, with a limit of detection (LOD) of 0.1 µg·kg−1 and recoveries of 87.2%–114.3%, within 10 min. The results showed good correlation (R2 > 0.99) between time-resolved fluorescent immunochromatographic strip test and high performance liquid chromatography (HPLC). Soybean sauce samples analyzed using time-resolved fluorescent immunochromatographic strip test revealed that 64.2% of samples contained aflatoxin B1 at levels ranging from 0.31 to 12.5 µg·kg−1. The strip test is a rapid, sensitive, quantitative, and cost-effective on-site screening technique in food safety analysis. PMID:27428975

  9. Monitoring the production of aflatoxin B1 in wheat by measuring the concentration of nor-1 mRNA.

    Science.gov (United States)

    Mayer, Zsuzsanna; Färber, Paul; Geisen, Rolf

    2003-02-01

    A real-time reverse transcription-PCR system has been used to monitor the expression of an aflatoxin biosynthetic gene of Aspergillus flavus in wheat. Therefore, total RNA was isolated from infected wheat samples, reverse transcribed and subjected to real-time PCR. In parallel all samples were analyzed by high-pressure liquid chromatography for aflatoxin B(1) production. The primer-probe system of the real-time PCR was targeted against nor-1, a gene of the aflatoxin biosynthetic pathway. By application of this method the nor-1 transcription was quantified during the course of incubation. After 4 days of incubation nor-1 mRNA could be detected for the first time. The amount of nor-1 mRNA increased rapidly, and the maximum was achieved after 6 days. Then, starting very slowly, the mRNA was degraded until day 8, and this was followed by a very fast degradation, reaching nondetectable levels at days 9 and 10. First traces of aflatoxin B(1)could be detected between the 5th and 6th day of incubation. The aflatoxin concentration reached its maximum after 9 days of incubation and remained constant for the whole period of observation. To ensure that differences in the nor-1 mRNA concentration were due to different expression levels, the expression of the constitutively expressed beta-tubulin gene (benA56) has also been monitored. The expression of benA56 remained constant during the whole incubation time. As a parameter for fungal growth, the number of nor-1 gene copies was determined during the course of incubation. The numbers of nor-1 gene copies increased at the beginning of the incubation and reached a plateau at day 5. They correlate well with the viable counts albeit at a higher level. PMID:12571042

  10. GSTM1 and XRCC3 Polymorphisms: Effects on Levels of Aflatoxin B1-DNA Adducts

    Institute of Scientific and Technical Information of China (English)

    Xi-dai Long; Yun Ma; Zhou-lin Deng

    2009-01-01

    Objective: Aflatoxin B1 (AFB1), which can cause the formation of AFB1-DNA adducts, is a known human carcinogen. AFB1-exposure individuals with inherited susceptible carcinogen-metabolizing or repairing genotypes may experience an increased risk of genotoxicity. This study was designed to investigate whether the polymorphisms of two genes, the metabolic gene Glutathione S-transferase M1 (GSTM1) and DNA repair gene x-ray repair cross-complementing group 3 (XRCC3), can affect the levels of AFB1-DNA adducts in Guangxi Population (n= 966) from an AFB1-exposure area.Methods: AFB1-DNA adducts were measured by ELISA, and GSTM1 and XRCC3 codon 241 genotypes were identified by PCR-RFLP.Results: The GSTM1-null genotype [adjusted odds ratio (OR) = 2.09; 95% confidence interval (CI) = 1.61(2.71] and XRCC3 genotypes with 241 Met alleles [i.e., XRCC3-TM and -MM, adjusted ORs (95% CI) were 1.43 (1.08(1.89) and 2.42 (1.13(5.22), respectively] were significantly associated with higher levels of AFB1-DNA adducts. Compared with those individuals who did not express any putative risk genotypes as reference (OR = 1), individuals featuring all of the putative risk genotypes did experience a significantly higher DNA-adduct levels (adjusted ORs were 2.87 for GSTM1-null and XRCC3-TM; 5.83 for GSTM1-null and XRCC3-MM). Additionally, there was a positive joint effect between XRCC3 genotypes and long-term AFB1 exposure in the formation of AFB1-DNA adducts.Conclusion: These results suggest that individuals with susceptible genotypes GSTM1-null, XRCC3-TM, or XRCC3-MM may experience an increased risk of DNA damage elicited by AFB1 exposure.

  11. Protective Roles of Sodium Selenite against Aflatoxin B1-Induced Apoptosis of Jejunum in Broilers

    Directory of Open Access Journals (Sweden)

    Xi Peng

    2014-12-01

    Full Text Available The effects of aflatoxin B1 (AFB1 exposure and sodium selenite supplementation on cell apoptosis of jejunum in broilers were studied. A total of 240 one-day-old male AA broilers were randomly assigned four dietary treatments containing 0 mg/kg of AFB1 (control, 0.3 mg/kg AFB1 (AFB1, 0.4 mg/kg supplement Se (+ Se and 0.3 mg/kg AFB1 + 0.4 mg/kg supplement Se (AFB1 + Se, respectively. Compared with the control broilers, the number of apoptotic cells, the expression of Bax and Caspase-3 mRNA were significantly increased, while the expression of Bcl-2 mRNA and the Bcl-2/Bax ratio were significantly decreased in AFB1 broilers. The number of apoptotic cells and the expression of Caspase-3 mRNA in AFB1 + Se broilers were significantly higher than those in the control broilers, but significantly lower than those in AFB1 broilers. There were no significant changes in the expression of Bax mRNA between AFB1 + Se and control broilers; the expression of Bcl-2 mRNA and the Bcl-2/Bax ratio in AFB1 + Se broilers were significantly lower than those in the control broilers, but significantly higher than those in AFB1 broilers. In conclusion, 0.3 mg/kg AFB1 in the diet can increase cell apoptosis, decrease Bcl-2 mRNA expression, and increase of Bax and Caspase-3 mRNA expression in broiler’s jejunum. However, supplementation of dietary sodium selenite at the concentration of 0.4 mg/kg Se may ameliorate AFB1-induced apoptosis by increasing Bcl-2 mRNA expression, and decreasing Bax and Caspase-3 mRNA expression.

  12. Aflatoxin B1 Degradation by Stenotrophomonas Maltophilia and Other Microbes Selected Using Coumarin Medium

    Directory of Open Access Journals (Sweden)

    Tiangui Niu

    2008-08-01

    Full Text Available Aflatoxin B1 (AFB1 is one of the most harmful mycotoxins in animal production and food industry. A safe, effective and environmentally sound detoxification method is needed for controlling this toxin. In this study, 65 samples were screened from various sources with vast microbial populations using a newly developed medium containing coumarin as the sole carbon source. Twenty five single-colony bacterial isolates showing AFB1 reduction activity in a liquid culture medium were selected from the screen. Isolate 35-3, obtained from tapir feces and identified to be Stenotrophomonas maltophilia, reduced AFB1 by 82.5% after incubation in the liquid medium at 37 °C for 72 h. The culture supernatant of isolate 35-3 was able to degrade AFB1 effectively, whereas the viable cells and cell extracts were far less effective. Factors influencing AFB1 degradation by the culture supernatant were investigated. Activity was reduced to 60.8% and 63.5% at 20 °C and 30 °C, respectively, from 78.7% at 37 °C. The highest degradation rate was 84.8% at pH 8 and the lowest was only 14.3% at pH 4.0. Ions Mg2+ and Cu2+ were activators for AFB1 degradation, howeverï��Œion Zn2+ was a strong inhibitor. Treatments with proteinase K, proteinase K plus SDS and heating significantly reduced or eradicated the degradation activity of the culture supernatant. The results indicated that the degradation of AFB1 by S. maltophilia 35-3 was enzymatic and could have a great potential in industrial applications.

  13. Aflatoxicosis: Lessons from Toxicity and Responses to Aflatoxin B1 in Poultry

    Directory of Open Access Journals (Sweden)

    Melissa S. Monson

    2015-09-01

    Full Text Available This review is a comprehensive introduction to the effects of poultry exposure to the toxic and carcinogenic mycotoxin aflatoxin B1 (AFB1. The relationship between AFB1 sensitivity and metabolism, major direct and indirect effects of AFB1, recent studies of gene expression and transcriptome responses to exposure, and mitigation strategies to reduce toxicity are discussed. Exposure to AFB1 primarily occurs by consumption of contaminated corn, grain or other feed components. Low levels of residual AFB1 in poultry feeds can cause reduction in growth, feed conversion, egg production, and compromised immune functions, resulting in significant economic costs to producers. Thus, AFB1 acts as a “force multiplier” synergizing the adverse effects of microbial pathogens and other agents, and factors detrimental to poultry health. Domestic turkeys (Meleagris gallopavo are one of the most sensitive animals known to AFB1 due, in large part, to a combination of efficient hepatic bioactivation by cytochromes P450 1A5 and 3A37, and deficient hepatic glutathione-S-transferase (GST-mediated detoxification. Because of their sensitivity, turkeys are a good model to investigate chemopreventive treatments and feed additives for their ability to reduce AFB1 toxicity. Transcriptome analysis (RNA-seq of turkey poults (liver and spleen has identified AFB1-induced gene expression changes in pathways of apoptosis, carcinogenesis, lipid regulation, antimicrobial activity, cytotoxicity and antigen presentation. Current research focuses on further identifying the molecular mechanisms underlying AFB1 toxicity with the goal of reducing aflatoxicosis and improving poultry health.

  14. Protective effect of pretreatment with thymoquinone against Aflatoxin B1 induced liver toxicity in mice

    Directory of Open Access Journals (Sweden)

    A Nili-Ahmadabadi

    2011-10-01

    Full Text Available "n  Background and the purpose of the study: Thymoquinone (TQ is one of the active components of Nigella sativa. The plant has been used in herbal medicine for treatment of many diseases including liver complications. The present study aimed to investigate protective effects of TQ on Aflatoxin B1 (AFB1 induced liver toxicity in mice. "n  Methods: Animals were divided into six groups and treated intraperitoneally. Group 1 (blank served as vehicle, group 2 (positive control received AFB1, Group 3 was treated with 9 mg/kg of TQ, Groups 4, 5 and 6 were treated with 4.5, 9 and 18 mg/kg of TQ, respectively. After three consecutive days, except for groups 1 and 3, animals were administered with a single dose of AFB1 (2 mg/kg. All the animals were killed 24 hrs following the AFB1 administration under ether anesthesia. Biochemical parameters including AST, ALT and ALP in serum samples and glutathione (GSH and malondialdehyde (MDA contents in liver homogenates were determined. Liver sections were collected for histopathological examination. "n  Results: Findings of this study showed that AST, ALT, ALP and MDA levels were significantly lower in the TQ treated animals as compared to AFB1 group (group 2. Furthermore, TQ was able to recover glutathione content (GSH of liver tissue. The best response, however, was observed with the dose of 9 mg/kg. Liver sections of AFB1 intoxicated mice showed inflammation, necrosis, hyperplasia of kupffer and infiltration of mononuclear cells, dilation of sinusoids and disruption of hepatocytes, while treatment with TQ helped to normalize liver architecture in accordance to biochemical findings. "n  Conclusion: Taken collectively, TQ has a protective role with optimum dose of 9 mg/kg in AFB1 hepatotoxicity.

  15. Effect of dietary resveratrol in ameliorating aflatoxin B1-induced changes in broiler birds.

    Science.gov (United States)

    Sridhar, M; Suganthi, R U; Thammiaha, V

    2015-12-01

    Consumption of aflatoxin B1 (AFB1) contaminated feed by poultry affects the health of broiler birds causing severe economic losses. The use of phytochemicals is a safe, effective, alternative and practical approach to combat the toxic effect of AF in broilers. Resveratrol, a polyphenol derived from red grapes, berries and peanuts, exerts anti-inflammatory, antioxidant and immunomodulatory effects. Our study was aimed at evaluating the possible protective effects of resveratrol against the adverse effects of AFB1 in broiler birds. A feeding trial of 42 days of duration was undertaken in a completely randomized design with five dietary treatments: G1-AFB1(1.0 ppm); G2-CTR (basal diet alone); G3-AFB1(1.0 ppm)+Resv 0.5%; G4-AFB1(1.0 ppm)+Resv 1%; and G5-Resv 1%. Gain in body weight (BWG) and feed intake (FI) was observed to be highest (p Feed conversion ratio was lowest in G2-CTR birds and failed to record any significant variation (p > 0.05) between groups as well as within groups. Birds fed resveratrol at both 0.5% and 1.0% levels in combination with AFB1 as well as alone along with basal diet had lower BWG and FI between the fourth and fifth week and also at the fifth week (p 0.05) was obtained in the FCR of AFB1 and resveratrol group of broiler birds. AFB1 feeding significantly increased the activities of aspartate-(AST) and alanine-(ALT) amino transferase, superoxide dismutase (SOD) and catalase (CAT) activities (p feed additive to control aflatoxicosis in poultry farms. PMID:25319220

  16. Square wave cathodic stripping voltammetric technique for determination of Aflatoxin B1 in ground nut sample

    International Nuclear Information System (INIS)

    An electro analytical method has been developed for the detection and determination of 2,3,6a,9a-tetrahydro-4-methoxy cyclo penta[c] furo[3, 2:4,5] furo [2,3-h][l] benzopyrane-1,11-dione (aflatoxin B1, AFB1) by a square wave cathodic stripping voltammetric (SWSV) technique on a hanging mercury drop electrode (HMDE) in aqueous solution with Britton-Robinson Buffer (BRB) at pH 9.0 as the supporting electrolyte. Effect of instrumental parameters such as accumulation potential (Eacc), accumulation time (tacc), scan rate (v), square wave frequency, step potential and pulse amplitude were examined. The best condition were found to be Eacc of -0.8 V, tacc of 100 s, v of 3750 mVs-1, frequency of 125 Hz, voltage step of 30 mV and pulse amplitude of 50 mV. Calibration curve was linear in the range of 0.01 to 0.15 μM with a detection limit of 0.125 x 10-8 M. Relative standard deviation for a replicate measurement of AFB1 (n = 5) with a concentration of 0.01 μM was 0.83 % with a peak potential of -1.30 V (against Ag/ AgCl). The recovery values obtained in spiked ground nut elute sample were 94.00 ± 0.67 % for 3.0 ppb, 91.22 ± 1.56 % for 9 ppb and 92.56 ± 2.00 % for 15.0 ppb of AFB1. The method was applied for the determination of the AFB1 in ground nut samples after extraction and clean-up steps. The results were compared with that obtained by high performance liquid chromatography (HPLC) technique. (author)

  17. Hepatitis B virus x gene and cyanobacterial toxins promote aflatoxin B1-induced hepatotumorigenesis in mice

    Institute of Scientific and Technical Information of China (English)

    Min Lian; Ying Liu; Shun-Zhang Yu; Geng-Sun Qian; Shu-Guang Wan; Kenneth R Dixon

    2006-01-01

    AIM: To assess the combinative role of aflatoxin B1 (AFB1),cyanobacterial toxins (cyanotoxins), and hepatitis B virus (HBV) x gene in hepatotumorigenicity.METHODS: One-week-old animals carrying HBV x gene and their wild-type littermates were intraperitoneally (ip) injected with either single-dose AFB1 [6 mg/kg body weight (bw)], repeated-dose cyanotoxins (microcystinLR or nodularin, 10 μg/kg bw oncea week for 15 wk),DMSO (vehicle control) alone, or AFB1 followed by cyanotoxins a week later, and were sacrificed at 24 and 52 wk post-treatment.RESULTS: AFB1 induced liver tumors in 13 of 29 (44.8%) transgenic mice at 52 wk post-treatment, significantly more frequent than in wild-type mice (13.3%). This significant difference was not shown in the 24-wk study. Compared with AFB1 exposure alone, MC-LR and nodularin yielded approximately 3-fold and 6-fold increases in the incidence of AFB1-induced liver tumors in wild-type animals at 24 wk, respectively. HBV x gene did not further elevate the risk associated with coexposure to AFB1 and cyanotoxins. With the exception of an MC-LR-dosed wild-type mouse, no liver tumor was observed in mice treated with cyanotoxins alone at 24 wk. Neither DMSO-treated transgenic mice nor their wild-type littermates had pathologic alterations relevant to hepatotumorigenesis in even up to 52 wk.CONCLUSION: HBV x gene and nodularin promote the development of AFB1-induced liver tumors. Co-exposure to AFB1 and MC-LR tends to elevate the risk of liver tumors at 24 wk relative to exposure to one of them.The combinative effect of AFB1, cyanotoxins and HBVx on hepatotumorigenesis is weak at 24 wk.

  18. Antigenotoxic Effect of Piperine in Broiler Chickens Intoxicated with Aflatoxin B1

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    Verônica da Silva Cardoso

    2016-10-01

    Full Text Available Piperine is an abundant amide extracted from black pepper seeds which has been shown to have protective effects against cytotoxic and genotoxic carcinogenesis induced by certain chemical carcinogens and aflatoxin B1 (AFB1 in vitro. The aim of this work was to study, in vivo, the antigenotoxic potential of feed-added piperine on broiler chickens experimentally intoxicated with AFB1, using micronucleus and comet assays. The antigenotoxicity assessment of 9-day-old chicks was performed on a total of 60 chickens divided into four groups of 15 broilers each: (C control, (P 60 mg·piperine kg−1 feed, (A 0.5 mg·AFB1·kg−1 body weight, (daily by oral route, and (P + A co-treatment with piperine and AFB1. The experiment was conducted for 26 days. Chicks intoxicated with AFB1 showed significant genotoxic effects in the first 24 h post intoxication, and the effects remained in the other periods analyzed (48, 72, and 96 h and 26 days of treatment. The DNA damage in peripheral blood cells, the number of erythrocytes with micronuclei, and polychromatic-to-normochromatic erythrocyte ratio were significantly reduced or absent in the piperine/AFB1 group. No significant differences were observed between the group piperine/AFB1 and the control and piperine-alone groups. The addition 60 mg·kg−1 of piperine to the diet of the broiler chicks was safe, promoting beneficial effects in poultry health with respect to the toxic effects 0.5 mg·AFB1·kg−1 body weight.

  19. 黄曲霍毒素B1人工抗原的合成及鉴定%Synthesis and Identification of Aflatoxin B1 Artificial Antigen

    Institute of Scientific and Technical Information of China (English)

    伍鑫茹; 杨雪娇; 赵肃清; 张焜

    2013-01-01

    Carboxyl group was introduced to Aflatoxin B1 (aflatoxin B1, AFB1) by derivatization method to synthesis the hapten (AFB1 carboxymethyl activator). And the AFBiO was coupled with BSA by using N-hydroxy-succinimide method to prepare the complete antigen of AFB,. The results of ECI-MS and ultraviolet spectroscopy showed that the target hapten was successfully synthesized. Through combined with ultraviolet spectrophotometry and regression equation, the standard curves of the different concentration hapten and BSA were abtained as follow: y=0.1440x+0.0103 (R2 =0.9986) and y=0.0059x+0.0808 (R2 =0.9889) respectively. The concentrations of AFB1O and BSA in adduct were 186.32 μg/mlL and 6127.46 ng/mL respectively, and the molar ration was 5.13:1.%采用衍生化在黄曲霉毒素B1(aflatoxin B1,AFB1)上引入羧基合成AFB1羧甲基活化物,通过N-羟琥珀酰亚胺酯(N-hydroxy-succinimide NHS)法将AFB1O与牛血清白蛋白(BSA)偶联,制备黄曲霉毒素B1完全抗原AFB1-BSA.ECI-MS和紫外光谱法的鉴定结果表明目标半抗原合成成功.结合紫外分光光度法和回归方程,分别测得不同浓度的半抗原和蛋白质线性曲线为:Y=0.1440X+0.0103,R2=0.9986; Y=0.0059X+0.0808,R2=0.9889.偶联物中半抗原和蛋白质的浓度分别为186.32 μg/mL、6127.46 μg/mL,即求得抗原分子结合比为5.13∶1,从而为制备抗AFB1抗体奠定基础.

  20. 茶叶中黄曲霉毒素B1的检测方法研究%Determination methods of aflatoxin B1 in tea

    Institute of Scientific and Technical Information of China (English)

    吴国华; 赵榕; 里南; 王雄

    2014-01-01

    Objectives To compare the three different methods for the determination of aflatoxin B1 in tea samples, including gold immune chromatography assay (GICA), high performance liquid chromatography (HPLC) and enzyme-linked immunosorbent assay (ELISA). Methods Method validation was done based on black tea, green tea and jasmine tea. And black tea, green tea, oolong tea, jasmine tea, extracted tea, medicinal health tea as representatives were detected with different methods. Results Both of optimized GICA and HPLC achieved high accuracy and precision. ELISA assay produced fake positive. Only one of eleven tested samples had aflatoxin B1 in random test.%目的:针对茶叶中黄曲霉毒素 B1的检测,对比胶体金免疫层析法、高效液相色谱、酶联免疫法的差异。方法以红茶、绿茶、花茶为基质,进行方法验证。并选取具有代表性的红茶、绿茶、乌龙茶、及花茶、萃取茶、药用保健茶分别用不同的方法检测。结果改良后的液相法和胶体金免疫层析法准确度和精密度较高,酶联免疫法存在假阳性。随机检测11种茶叶仅有一种检出黄曲霉毒素B1

  1. Degradation and Application of Aflatoxin B1 by Aspergillus Niger%黑曲霉对黄曲霉毒素B1的降解与应用研究

    Institute of Scientific and Technical Information of China (English)

    李冰; 董征英; 常维山

    2012-01-01

    试验利用黑曲霉对黄曲霉毒素B1进行了降解率、活性组分的确定、安全性及对饲料中降解黄曲霉毒素效果的研究。研究结果表明,黑曲霉对黄曲霉毒素B1的降解率达93.28%,其中黑曲霉的胞外粗提液对黄曲霉毒素B1的降解活性最高,证明黑曲霉对黄曲霉毒素B1的降解是一种生物化学反应,黑曲霉发酵液对饲料中的黄曲霉毒素B1具有很强的降解作用。小鼠的急性毒理学试验证明,试验用黑曲霉本身具有良好的安全性。%The determination of degradation efficiency and active components of aflatoxin B1 safety and the effect of degradation aflatoxin in feed about the screened aspergillus was researched in this experiment used aspergillus niger. The result showed that the degradation of aflatoxin B1 by aspergillus niger was 93.28%, the degrading efficiencies of aflatoxin B1 by exocellular coarse extraction liquid was highest, and this proved that the degration of aflatoxin B1 by aspergillus niger was a biological reaction. The degrading effect of aflatoxin B1 in feed by aspergillus fermented liquid was strong. A favorable safety of aspergillus niger was testified through the acute toxicology experiments in mice.

  2. Inhibitory Effect of Cinnamaldehyde, Citral, and Eugenol on Aflatoxin Biosynthetic Gene Expression and Aflatoxin B1 Biosynthesis in Aspergillus flavus.

    Science.gov (United States)

    Liang, Dandan; Xing, Fuguo; Selvaraj, Jonathan Nimal; Liu, Xiao; Wang, Limin; Hua, Huijuan; Zhou, Lu; Zhao, Yueju; Wang, Yan; Liu, Yang

    2015-12-01

    In order to reveal the inhibitory effects of cinnamaldehyde, citral, and eugenol on aflatoxin biosynthesis, the expression levels of 5 key aflatoxin biosynthetic genes were evaluated by real-time PCR. Aspergillus flavus growth and AFB1 production were completely inhibited by 0.80 mmol/L of cinnamaldehyde and 2.80 mmol/L of citral. However, at lower concentration, cinnamaldehyde (0.40 mmol/L), eugenol (0.80 mmol/L), and citral (0.56 mmol/L) significantly reduced AFB1 production with inhibition rate of 68.9%, 95.4%, and 41.8%, respectively, while no effect on fungal growth. Real-time PCR showed that the expressions of aflR, aflT, aflD, aflM, and aflP were down-regulated by cinnamaldehyde (0.40 mmol/L), eugenol (0.80 mmol/L), and citral (0.56 mmol/L). In the presence of cinnamaldehyde, AflM was highly down-regulated (average of 5963 folds), followed by aflP, aflR, aflD, and aflT with the average folds of 55, 18, 6.5, and 5.8, respectively. With 0.80 mmol/L of eugenol, aflP was highly down-regulated (average of 2061-folds), followed by aflM, aflR, aflD, and aflT with average of 138-, 15-, 5.2-, and 4.8-folds reduction, respectively. With 0.56 mmol/L of citral, aflT was completely inhibited, followed by aflM, aflP, aflR, and aflD with average of 257-, 29-, 3.5-, and 2.5-folds reduction, respectively. These results suggest that the reduction in AFB1 production by cinnamaldehyde, eugenol, and citral at low concentration may be due to the down-regulations of the transcription level of aflatoxin biosynthetic genes. Cinnamaldehyde and eugenol may be employed successfully as a good candidate in controlling of toxigenic fungi and subsequently contamination with aflatoxins in practice.

  3. Rapid and simple RIA technique of the determining aflatoxin B1 in cereals and fodder using commercially available kits

    International Nuclear Information System (INIS)

    A new sample extraction procedure was tested for facilitating practical large-scale application of the method of RIA testing cereal and fodder contamination with aflatoxin B1. While in the conventional procedure, chloroform extracts are prepared and the solvent has to be removed by evaporation, in the procedure suggested the samples are extracted with methanol or acetone and the extracts are only diluted with a buffer prior to radioimmunoassay. As compared to the former approach, the new method exhibits a commensurable aflatoxin recovery (83 to 90% for concentrations of 2 to 20 μg/kg) and a poorer detection limit (1 μg/kg as against the conventional 0.2 μg/kg), the results, however, are appreciably less affected by interferents and the procedure is considerably simpler. (author). 2 tabs., 9 refs

  4. Vaccination of heifers with anaflatoxin improves the reduction of aflatoxin b1 carry over in milk of lactating dairy cows.

    Directory of Open Access Journals (Sweden)

    Laura Giovati

    Full Text Available It was previously reported that injection of anaflatoxin B1 (AnAFB1 conjugated to keyhole limpet hemocyanin (KLH, together with Freund's adjuvant, was effective in inducing in cows a long lasting titer of anti-aflatoxin B1 (AFB1 antibodies (Abs, cross-reacting with other aflatoxins, which were able to hinder, proportionally to their titer, the secretion of aflatoxin M1 (AFM1 into the milk of cows continuously fed with AFB1. According to anti-AFB1 Ab titer, 50% of the vaccinated cows were recognized as high responder animals. In an attempt to prepare a more effective formulation for vaccination of cows, it was compared the immunogenicity, in Holstein Friesian heifers, of AnAFB1 covalently conjugated to KLH or to recombinant diphtheria toxin (CRM197 molecules, and injected together with various adjuvants. This study demonstrated that injection of AnAFB1 conjugated to KLH and mixed with complete (priming and incomplete Freund's adjuvant (boosters, as in the previous schedule of immunization, was the most effective regimen for inducing Ab responses against AFB1, although pre-calving administration could increase the effectiveness of vaccination, resulting in 100% high responder animals. After one booster dose at the beginning of the milk production cycle, anti-AFB1 Ab titers were comparable to those recorded at the end of the immunization schedule, and proved to be effective in reducing significantly AFB1 carry over, as AFM1, from feed to milk. Pre-calving vaccination of dairy heifers with conjugated AnAFB1, adjuvated with complete and incomplete Freund's adjuvant, may represent the most effective tool for preventing the public health hazard constituted by milk and cheese contaminated with aflatoxins.

  5. Vaccination of Heifers with Anaflatoxin Improves the Reduction of Aflatoxin B1 Carry Over in Milk of Lactating Dairy Cows

    Science.gov (United States)

    Giovati, Laura; Gallo, Antonio; Masoero, Francesco; Cerioli, Carla; Ciociola, Tecla; Conti, Stefania; Magliani, Walter; Polonelli, Luciano

    2014-01-01

    It was previously reported that injection of anaflatoxin B1 (AnAFB1) conjugated to keyhole limpet hemocyanin (KLH), together with Freund's adjuvant, was effective in inducing in cows a long lasting titer of anti-aflatoxin B1 (AFB1) antibodies (Abs), cross-reacting with other aflatoxins, which were able to hinder, proportionally to their titer, the secretion of aflatoxin M1 (AFM1) into the milk of cows continuously fed with AFB1. According to anti-AFB1 Ab titer, 50% of the vaccinated cows were recognized as high responder animals. In an attempt to prepare a more effective formulation for vaccination of cows, it was compared the immunogenicity, in Holstein Friesian heifers, of AnAFB1 covalently conjugated to KLH or to recombinant diphtheria toxin (CRM197) molecules, and injected together with various adjuvants. This study demonstrated that injection of AnAFB1 conjugated to KLH and mixed with complete (priming) and incomplete Freund's adjuvant (boosters), as in the previous schedule of immunization, was the most effective regimen for inducing Ab responses against AFB1, although pre-calving administration could increase the effectiveness of vaccination, resulting in 100% high responder animals. After one booster dose at the beginning of the milk production cycle, anti-AFB1 Ab titers were comparable to those recorded at the end of the immunization schedule, and proved to be effective in reducing significantly AFB1 carry over, as AFM1, from feed to milk. Pre-calving vaccination of dairy heifers with conjugated AnAFB1, adjuvated with complete and incomplete Freund's adjuvant, may represent the most effective tool for preventing the public health hazard constituted by milk and cheese contaminated with aflatoxins. PMID:24714096

  6. Mechanisms of butylated hydroxytoluene chemoprevention of aflatoxicosis-inhibition of aflatoxin B1 metabolism

    International Nuclear Information System (INIS)

    Chemoprevention of toxicoses and/or cancer through the use of nutrients or pharmacologic compounds is the subject of intense study. Among the many compounds examined, food additives such as antioxidants are being considered due to their ability to reduce disease formation by either induction or inhibition of key enzyme systems. One such compound, butylated hydroxytoluene (BHT), has been found to protect against cancer formation caused by exposure to aflatoxin B1 (AFB1) in rodents. We have shown that dietary BHT protects against clinical signs of aflatoxicosis in turkeys, a species that is very susceptible to this mycotoxin. In this study, the effect of BHT on AFB1 metabolism and other cytochrome P450 (CYP)-related enzyme activities in turkey liver microsomes was examined to discern possible mechanisms of BHT-mediated protection against aflatoxicosis. Ethoxyresorufin O-deethylase (EROD), methoxyresorufin O-demethylase (MROD), prototype activities for CYP1A1 and 1A2, respectively, were decreased in the BHT fed (4000 ppm) animals, while oxidation of nifedipine, a prototype activity for CYP3A4, was increased. However, BHT added to microsomal incubations inhibited these CYP activities in a concentration-related manner. Importantly, BHT inhibited conversion of AFB1 to the reactive intermediate AFB1-8,-9-epoxide (AFBO), exhibiting Michaelis-Menton competitive inhibition kinetics (Ki = 0.81 μM). Likewise, microsomes prepared from turkeys fed BHT were significantly less active in AFBO formation compared to those from control birds. When turkeys were fed BHT for up to 40 days, residual BHT was present in liver, breast meat, thigh meat and abdominal fat in concentrations substantially below U.S. FDA guidelines for this antioxidant, but in concentrations greater than the Ki, likely sufficient to inhibit bioactivation of AFB1in vivo. BHT-induced hydropic degeneration in the livers of BHT fed animals was significantly greater in birds that remained on BHT treatment for up to 30

  7. Effect of the inclusion of adsorbents on aflatoxin B1 quantification in animal feedstuffs.

    Science.gov (United States)

    Gallo, A; Masoero, F; Bertuzzi, T; Piva, G; Pietri, A

    2010-01-01

    The extraction efficiency of aflatoxin B1 (AFB1) in cattle feed containing nine adsorbents (ADSs) was investigated using two organic/aqueous solvents composed of methanol/water (80/20 v/v; MeOH) and acetone/water (85/15 v/v; AC). Samples were obtained including a highly AFB1-contaminated (HC) and a low-level AFB(1)-contaminated (LC) feedstuff (15.33 and 7.57 microg kg(-1), respectively), nine ADSs (four clay minerals; one yeast cell wall-based product; one activated carbon and three commercial ADS products) at two different levels of inclusion (10 and 20 g kg(-1)). After solvent extraction and immunoaffinity column clean-up, all samples were analysed for AFB1 by high-performance liquid chromatography (HPLC) with fluorescence detection. For each contamination level (HC and LC), the data obtained were analysed using a factorial arrangement in a completely randomized design. Means were compared with the correspondent controls using the Dunnett's test. No statistical difference was found in AFB1 levels of feedstuffs not containing ADSs when extracted with AC or MeOH, even if numerically higher values were obtained with AC. A dose-dependent effect (p < 0.01) of ADSs inclusion was observed on AFB1 recoveries that were lower when the higher ADS level (20 g kg(-1)) was included in the HC and LC feedstuffs. Higher AFB(1) recoveries were obtained using AC compared with MeOH, both in HC (75.0% versus 12.0%, respectively) and in LC (84.0% versus 22.8%, respectively) ADSs containing feedstuffs. However, when the activated carbon and the sodium bentonite were included in feeds, lower AFB1 concentrations with respect to control values (p < 0.001 and <0.05, respectively) were obtained also using AC. The data obtained in this study indicate that routine use of the MeOH solvent for AFB1 analysis of unknown feedstuffs, can produce misleading results if they contain an ADS. PMID:19750400

  8. Leaky Gut and Mycotoxins: Aflatoxin B1 Does Not Increase Gut Permeability in Broiler Chickens

    Science.gov (United States)

    Galarza-Seeber, Rosario; Latorre, Juan D.; Bielke, Lisa R.; Kuttappan, Vivek A.; Wolfenden, Amanda D.; Hernandez-Velasco, Xochitl; Merino-Guzman, Ruben; Vicente, Jose L.; Donoghue, Annie; Cross, David; Hargis, Billy M.; Tellez, Guillermo

    2016-01-01

    Previous studies conducted in our laboratory have demonstrated that intestinal barrier function can be adversely affected by diet ingredients or feed restriction, resulting in increased intestinal inflammation-associated permeability. Two experiments were conducted in broilers to evaluate the effect of three concentrations of Aflatoxin B1 (AFB1; 2, 1.5, or 1 ppm) on gastrointestinal leakage and liver bacterial translocation (BT). In experiment 1, 240 day-of-hatch male broilers were allocated in two groups, each group had six replicates of 20 chickens (n = 120/group): Control feed or feed + 2 ppm AFB1. In experiment 2, 240 day-of-hatch male broilers were allocated in three groups, each group had five replicates of 16 chickens (n = 80/group): Control feed; feed + 1 ppm AFB1; or feed + 1.5 ppm AFB1. In both experiments, chickens were fed starter (days 1–7) and grower diets (days 8–21) ad libitum and performance parameters were evaluated every week. At day 21, all chicks received an oral gavage dose of FITC-d (4.16 mg/kg) 2.5 h before collecting blood samples to evaluate gastrointestinal leakage of FITC-d. In experiment 2, a hematologic analysis was also performed. Liver sections were aseptically collected and cultured using TSA plates to determine BT. Cecal contents were collected to determine total colony-forming units per gram of Gram-negative bacteria, lactic acid bacteria (LAB), or anaerobes by plating on selective media. In experiment 2, liver, spleen, and bursa of Fabricius were removed to determine organ weight ratio, and also intestinal samples were obtained for morphometric analysis. Performance parameters, organ weight ratio, and morphometric measurements were significantly different between Control and AFB1 groups in both experiments. Gut leakage of FITC-d was not affected by the three concentrations of AFB1 evaluated (P > 0.05). Interestingly, a significant reduction in BT was observed in chickens that received 2 and

  9. Differential expression of genes during aflatoxin B1-induced hepatocarcinogenesis in tree shrews

    Institute of Scientific and Technical Information of China (English)

    Yuan Li; Dan Luo; Hui-Fen Yue; Li-Sheng Zhang; Jian-Ren Gu; Da-Fang Wan; Jian-Jia Su; Ji Cao; Chao Ou; Xiao-Kun Qiu; Ke-Chen Ban; Chun Yang; Liu-Liang Qin

    2004-01-01

    AIM: Through exploring the regulation of gene expression during hepatocarcinogenesis induced by aflatoxin B1 (AFB1),to find out the responsible genes for hepatocellular carcinoma (HCC) and to further understand the underlying molecular mechanism.METHODS: Tree shrews ( 7upaia belangeri chinensis)were treated with or without AFB1 for about 90 weeks. Liver biopsies were performed regularly during the animal experiment. Eight shares of total RNA were respectively isolated from 2 HCC tissues, 2 HCC-surrounding noncancerous liver tissues, 2 biopsied tissues at the early stage (30th week) of the experiment from the same animals as above, 1 mixed sample of three liver tissues biopsied at the beginning (0th week) of the experiment, and another 1 mixed sample of two liver tissues from the untreated control animals biopsied at the 90th week of the experiment. The samples were then tested with the method of AtlasTM cDNA microarray assay. The levels of gene expression in these tissues taken at different time points during hepatocarcinogenesis were compared.RESULTS: The profiles of differently expressed genes were quite different in different ways of comparison. At the same period of hepatocarcinogenesis, the genes in the same function group usually had the same tendency for up- or down-regulation. Among the checked 588 genes that were known to be related to human cancer, 89 genes (15.1%) were recognized as "important genes" because they showed frequent changes in different ways of comparison. The differentially expressed genes during hepatocarcinogenesis could be classified into four categories: genes up-regulated in HCC tissue, genes with similar expressing levels in both HCC and HCC-surrounding liver tissues which were higher than that in the tissues prior to the development of HCC,genes down-regulated in HCC tissue, and genes up-regulated prior to the development of HCC but down-regulated after the development of HCC.CONCLUSION: A considerable number of genes could change

  10. Effects of milk thistle seed against aflatoxin B 1 in broiler model

    Directory of Open Access Journals (Sweden)

    Halimeh Amiridumari

    2013-01-01

    Full Text Available Background: Consumption of aflatoxin B 1 (AFB 1 contaminated products can pose a risk of development of various diseases in human and animals due to radical production. The scope of this work is to evaluate the efficacy of milk thistle seed (MTS, as a radical scavenger, on serum biochemistry, lipid profile and liver enzymes against AFB 1 in broiler chickens contaminated with AFB 1 . Materials and Methods: The effect of nine experimental treatments (3 × 3 factorial design was assessed using 216 one-d-old Ross 308 male broiler chicks in a randomized complete design with four replicates of six birds for each dietary treatments: Control (T1, 250 ppb AFB 1 (T2, 500 ppb AFB 1 (T3, 0.5% MTS (T4, 0.5% MTS Plus 250 ppb AFB 1 (T5, 0.5% MTS Plus 500 ppb AFB1 (T6, 1.0% MTS (T7, 1.0% MTS Plus 250 ppb AFB 1 (T8, and 1.0% MTS Plus 500 ppb AFB 1 (T9. The individual and combined effects of dietary AFB 1 and MTS on serum biochemistry factors (Glucose, Calcium, Phosphorus, Iron, Creatinine, and Uric acid, lipid profile (Triglyceride, Cholesterol, Low density lipoprotein (LDL, and High density lipoprotein (HDL and liver enzymes aspartate amino-transferase and alanine amino-transaminase (ALT in broilers were evaluated at 21 days of age. Also, statistical packages Macros-1.002 (2010 were used to perform the above analysis on computer. Results: Consumption of 500 ppb AFB 1 in to the diet significantly decreased HDL (58.13 ± 2.65, Calcium (7.11 ± 0.13, and Glucose (197.1 ± 7.42 compared to the control group (85.12 ± 1.95, 9.45 ± 0.17 and 223.1 ± 6.61, respectively, (P < 0.05. In contrast, it significantly increased creatinine (2.25 ± 0.011 and AST (244.51 ± 4.91. Using MTS together with AFB 1 significantly reduced the effect of AFB 1 on the above parameters. Conclusion: MTS can provide protection against the negative effects of AFB 1 on broiler chicks.

  11. Novel regulation of aflatoxin B1 biosynthesis in Aspergillus flavus by piperonal.

    Science.gov (United States)

    Park, Eun-Sil; Bae, In Kyung; Kim, Ho Jin; Lee, Sung-Eun

    2016-08-01

    The present study investigated its inhibitory role in aflatoxin (AF) biosynthesis. Treating only AFB1- and B2-producing Aspergillus flavus with piperonal completely inhibited AFB1 production with high sclerotial formation, resulting in 20-fold higher AFG2 production. On the other hand, benzodioxole and eugenol suppressed AFB1 production without AFG formation, while methyleugenol showed potent inhibition of AFB1 production with slight production of AFG1. These results indicate that natural products may change aflatoxin biosynthesis, and highlight a novel regulation of AFG2 production by piperonal. It is the first report for chemical regulation on AFG2 production in non-AFG producing-aspergilli. PMID:26273991

  12. The effects of necrotic enteritis, aflatoxin B1, and virginiamycin on growth performance, necrotic enteritis lesion scores, and mortality in young broilers.

    Science.gov (United States)

    Cravens, R L; Goss, G R; Chi, F; De Boer, E D; Davis, S W; Hendrix, S M; Richardson, J A; Johnston, S L

    2013-08-01

    The effects of increasing aflatoxin B1 concentration (0, 0.75, 1.5 mg/kg) on broilers with or without necrotic enteritis or virginiamycin were determined. In the 23-d study, 22 male Cobb 500 chicks per pen were allotted to 12 treatments (3 × 2 × 2 factorial arrangement) with 8 replications. Intestines of 5 birds per pen were examined for lesions on d 21. Birds were allowed to consume feed and water ad libitum. Aflatoxin was included in the diets from d 0. All birds received a 10× dose of coccidiosis vaccine on d 10. Pens of birds where necrotic enteritis was being induced were on Clostridium perfringens pathogen (CPP) contaminated litter from d 0. Aflatoxin decreased gain and feed intake and resulted in poorer feed:gain, increased mortality, and higher lesion scores. Inducing necrotic enteritis increased lesion scores and decreased feed intake and gain. Adding virginiamycin to the diets improved gain, feed intake, feed conversion, and decreased mortality. There was a 3-way interaction (aflatoxin × virginiamycin × CPP) on gain; increasing aflatoxin decreased gain and the effects of CPP and virginiamycin were dependent on aflatoxin concentration. In the absence of aflatoxin virginiamycin increased gain but was unable to prevent the growth suppression caused by CPP. At 0.75 mg/kg of aflatoxin virginiamycin no longer increased growth in non-CPP challenged birds but was able to increase growth in CPP-challenged birds. At the 1.5 mg/kg of aflatoxin concentration, virginiamycin increased gain in non-CPP-challenged birds but challenging birds with CPP had no effect on gain. Virginiamycin improved overall feed conversion with the greatest improvement at 1.5 mg/kg (aflatoxin × virginiamycin, P enteritis decrease broiler performance and interact to decrease weight gain, virginiamycin helps improve gain in challenged birds at 0.75 mg/kg of aflatoxin, but not at 1.5 mg/kg of aflatoxin.

  13. Panax ginseng extract modulates oxidative stress, DNA fragmentation and up-regulate gene expression in rats sub chronically treated with aflatoxin B1 and fumonisin B 1.

    Science.gov (United States)

    Hassan, Aziza M; Abdel-Aziem, Sekena H; El-Nekeety, Aziza A; Abdel-Wahhab, Mosaad A

    2015-10-01

    Aflatoxins and fumonisins are important food-borne mycotoxins implicated in human health and have cytotoxic effects. The aims of the current study were to evaluate the protective role of Panax ginseng extract (PGE) against the synergistic effect of subchronic administration of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) on DNA and gene expression in rat. Female Sprague-Dawley rats were divided into eight groups (ten rats/group) and treated for 12 weeks including the control group, the group having received AFB1 (80 µg/kg bw), the group having received FB1 (100 µg/kg bw), the group having received AFB1 plus FB1 and the groups having received PGE (20 mg/kg bw) alone or with AFB1 and/or FB1. At the end of experiment, liver and kidney were collected for the determination of DNA fragmentation, lipid peroxidation (LP), glutathione (GSH) contents and alterations in gene expression. The results indicated that these mycotoxins increased DNA fragmentation, LP and decreased GSH content in liver and kidney and down-regulated gene expression of antioxidants enzymes. The combined treatments with AFB1 and/or FB1 plus PGE suppressed DNA fragmentation only in the liver, normalized LP and increased GSH in the liver and kidney as well as up-regulated the expression of GPx, SOD1 and CAT mRNA. It could be concluded that AFB1 and FB1 have synergistic genotoxic effects. PGE induced protective effects against their oxidative stress and genotoxicity through its antioxidant properties. PMID:24748134

  14. Determination Of The Aflatoxin B1 In Ground Nut By Differential Pulse Cathodic Stripping Voltammetry (Dpcsv) Technique

    OpenAIRE

    Yaacob, Mohammad Hadzri; Yusoff, Abdull Rahim Hj. Mohd.; Ahamad, Rahmalan

    2009-01-01

    An electro analytical method has been developed for the detection and determination of the 2,3,6a,9a-tetrahydro- 4-methoxycyclo penta[c] furo[3’,2’:4,5] furo [2,3-h][l] benzopyran-1,11-dione (aflatoxin B1, AFB1) by a differential pulse cathodic stripping voltammetry on a hanging mercury drop electrode (HMDE) in aqueous solution with Britton-Robinson buffer (BRb) as supporting electrolyte. Effect of instrumental parameters such as accumulation potential (Eacc), accumulation time (tacc) and ...

  15. Modulatory effects of essential oils from spices on the formation of DNA adduct by aflatoxin B1 in vitro.

    Science.gov (United States)

    Hashim, S; Aboobaker, V S; Madhubala, R; Bhattacharya, R K; Rao, A R

    1994-01-01

    Essential oils from common spices such as nutmeg, ginger, cardamom, celery, xanthoxylum, black pepper, cumin, and coriander were tested for their ability to suppress the formation of DNA adducts by aflatoxin B1 in vitro in a microsomal enzyme-mediated reaction. All oils were found to inhibit adduct formation very significantly and in a dose-dependent manner. The adduct formation appeared to be modulated through the action on microsomal enzymes, because an effective inhibition on the formation of activated metabolite was observed with each oil. The enzymatic modulation is perhaps due to the chemical constituents of the oils, and this could form a basis for their potential anticarcinogenic roles. PMID:8058527

  16. Analysis of Aflatoxin B1 in Iranian Foods Using HPLC and a Monolithic Column and Estimation of its Dietary Intake

    OpenAIRE

    Yazdanpanah, Hassan; Zarghi, Afshin; Shafaati, Ali Reza; FOROUTAN, SEYED MOHSEN; Aboul-Fathi, Farshid; Khoddam, Arash; Nazari, Firoozeh; Shaki, Fatemeh

    2013-01-01

    A high performance liquid chromatographic method was developed for determination of aflatoxin B1 (AFB1) in foods using a monolithic column with sample clean up on an immunoaffinity column. The method was validated for analysis of AFB1 in rice, bread, puffed corn snack, wheat flour and peanut samples. The average recoveries for AFB1 in different foods ranged from 94.4 to 102.5% with the coefficient of variation lower than 10% for all foods. Limit of detection was 0.01 ng/g. A survey of AFB1 wa...

  17. Effects of maternal exposure to aflatoxin B1 during pregnancy on fertility output of dams and developmental, behavioral and reproductive consequences in female offspring using a rat model.

    Science.gov (United States)

    Supriya, Ch; Akhila, B; Pratap Reddy, K; Girish, B P; Sreenivasula Reddy, P

    2016-01-01

    A suboptimal in utero environment can have detrimental effects on the pregnancy and long-term adverse "programing" effects on the offspring. Aflatoxin B1 is one of the potent reproductive toxicants and currently detected in both milk and tissues. This article focuses on the effects of prenatal exposure to graded doses of aflatoxin B1 on the pregnancy outcomes of dams and postnatal developments of the female offspring, since these issues have ethological relevance in both animals and humans. Pregnant Wistar rats were injected intramuscularly with vehicle or aflatoxin B1 (10, 20, 50 or 100 μg/kg body weight/day) on days 12-19 of gestation. At parturition, newborns were observed for clinical signs of toxicity and survival. The female offspring were examined through a battery of tests in order to evaluate their developmental, behavioral and reproductive end points. All animals were born alive. The litter size of the aflatoxin B1 treated rats was comparable to the controls. However, the birth weight of the pups in the experimental group was significantly lower when compared to controls. Significant and persistent lags in cliff avoidance, negative geotaxis, surface rightening activity and ascending wire mesh, with a delay in elapsed time for vaginal opening were detected in the female progeny exposed to aflatoxin B1 during embryonic development. The locomotor activity and exploratory behavior in experimental females were significantly decreased than that of controls. Embryonic exposure to aflatoxin B1 also resulted in prolonged stress response, irregular estrus and suppressed fertility output in the progeny at their adulthood. These results indicate that in utero exposure to aflatoxin B1 severely compromised postnatal development of neonatal rats and caused irregular estrus that was accompanied by suppressed fertility output. PMID:26956420

  18. Aflatoxin B1 residues in imported and local broiler, s breast and thigh muscle in Kurdistan region

    Directory of Open Access Journals (Sweden)

    E.P. Candlan

    2015-06-01

    Full Text Available Residues of Aflatoxins and their metabolites might be present in meat and other products of animals receiving Aflatoxin contaminated feeds which could subsequently create health problems in man. Eighty nine imported (Iran/Khosh pokht; (Turkey/Yam-tapilic, Lades, Senplic, Kapidac, Kozoa, Oznesilpilic and (Brazil, hilal, Sadia, and 90 locally produced (Hoshiar poultry farm, Nihad poultry farm, Hokar poultry farm, Mansoor poultry farm, AL-Shimal poultry house, Mardin poultry house and AL-Eetimad poultry slaughterhouse broiler breast and thigh muscle samples were examined for residual Aflatoxin B1 using ELIZA test. Results revealed that out of 89 imported samples only 21 (23.59% were positive, but only 2 (2.24% were rejected, while the remaining 87 samples (97.75% were acceptable. Concerning the local samples, results showed that 19 samples (21.11% were positive, but 10 (11.11% were rejected, while the remaining 80 samples (88.88% were accepted. The public health importance of residual AFB1 in broiler meat samples was discussed.

  19. Study on aflatoxin B1 detecting in sauce by colloidal gold immunochromatographic assay%胶体金免疫层析法检测酱油中黄曲霉毒素B1

    Institute of Scientific and Technical Information of China (English)

    赖卫华; 刘道峰; 邓省亮

    2012-01-01

    采用胶体金免疫层析法检测酱油中的黄曲霉毒素B1.加标的酱油样品经提取后,以胶体金免疫层析法对其进行黄曲霉毒素B1测定,并与酶联免疫吸附法进行比较.结果表明,当酱油中黄曲霉毒素含量超过国家限量标准(5 μg/kg),胶体金免疫层析法检测结果为阳性,说明该方法能够满足酱油样品中黄曲霉毒素B1监控的要求.%Colloidal gold immunochromatographic assay was used to detect the Aflatoxin B1 in Sauce. After spiking with aflatoxin B1 and extracting, the samples from sauce were detected by colloidal gold immunochromatographic assay and enzyme-linked immunosorbent assay. The data showed that the result of colloidal gold immunochromatographic assay would be positive when the concentration of aflatoxin B1 in sauce exceeding the maximum limit of China (5 jig/kg). The immunochromatographic assay can be used to screen aflatoxin B1 of sauce products.

  20. Non-Linear Relationships between Aflatoxin B1 Levels and the Biological Response of Monkey Kidney Vero Cells

    Directory of Open Access Journals (Sweden)

    Mendel Friedman

    2013-08-01

    Full Text Available Aflatoxin-producing fungi contaminate food and feed during pre-harvest, storage and processing periods. Once consumed, aflatoxins (AFs accumulate in tissues, causing illnesses in animals and humans. Most human exposure to AF seems to be a result of consumption of contaminated plant and animal products. The policy of blending and dilution of grain containing higher levels of aflatoxins with uncontaminated grains for use in animal feed implicitly assumes that the deleterious effects of low levels of the toxins are linearly correlated to concentration. This assumption may not be justified, since it involves extrapolation of these nontoxic levels in feed, which are not of further concern. To develop a better understanding of the significance of low dose effects, in the present study, we developed quantitative methods for the detection of biologically active aflatoxin B1 (AFB1 in Vero cells by two independent assays: the green fluorescent protein (GFP assay, as a measure of protein synthesis by the cells, and the microculture tetrazolium (MTT assay, as a measure of cell viability. The results demonstrate a non-linear dose-response relationship at the cellular level. AFB1 at low concentrations has an opposite biological effect to higher doses that inhibit protein synthesis. Additional studies showed that heat does not affect the stability of AFB1 in milk and that the Vero cell model can be used to determine the presence of bioactive AFB1 in spiked beef, lamb and turkey meat. The implication of the results for the cumulative effects of low amounts of AFB1 in numerous foods is discussed.

  1. Phenolic extract of Parkia biglobosa fruit pulp stalls aflatoxin B1 – mediated oxidative rout in the liver of male rats

    Directory of Open Access Journals (Sweden)

    Taofeek O. Ajiboye

    2014-12-01

    Full Text Available The effect of phenolic extract of Parkia biglobosa (Jacq. R. Br. ex G. Don, Fabaceae, pulp on aflatoxin B1 induced oxidative imbalance in rat liver was evaluated. Thirty-five male rats were randomized into seven groups of five animals each. Rats in group A served as control and received vehicle for drug administration (0.5% DMSO once daily at 24 h intervals for six weeks. Rats in groups B, D, E, F and G, received aflatoxin B1 (167 μg/kg body weight in 0.5% DMSO for three weeks, starting from the third week of the experimental period. Rats in Group C received 400 mg/kg bodyweight of the extract for six weeks, while groups D, E and F rats were treated with 100, 200 and 400 mg/kg bodyweight of the extract for six weeks respectively. Group G rats received 100 mg/kg body weight of vitamin C. Aflatoxin B1-mediated decrease in the activities of superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase and glucose-6-phosphate dehydrogenase were significantly attenuated. Aflatoxin B1 mediated the elevation in malondialdehyde, conjugated dienes, lipid hydroperoxides, protein carbonyl, and significantly lowered DNA fragmentation percentage. Overall, the phenolic extract of P. biglobosa pulp stalls aflatoxin B1-mediated oxidative rout by enhancing antioxidant enzyme activities leading to decreased lipid peroxidation, protein oxidation and DNA fragmentation.

  2. A multiplex chemiluminescent biosensor for type B-fumonisins and aflatoxin B1 quantitative detection in maize flour.

    Science.gov (United States)

    Zangheri, Martina; Di Nardo, Fabio; Anfossi, Laura; Giovannoli, Cristina; Baggiani, Claudio; Roda, Aldo; Mirasoli, Mara

    2015-01-01

    A multiplex chemiluminescent biosensor for simple, rapid and ultrasensitive on-site quantification of aflatoxin B1 and type B-fumonisins in maize samples has been developed. The biosensor integrates a multiplex indirect competitive lateral flow immunoassay (LFIA) based on enzyme-catalyzed chemiluminescence detection and a highly sensitive portable charge-coupled device (CCD) camera, employed in a lensless "contact" imaging configuration. The developed assay requires a simple extraction of the analytes from maize flour samples followed by their detection with a 30 min assay time. The use of chemiluminescence detection allowed accurate and objective analytes quantification, enabling simultaneous detection of type B-fumonisins and aflatoxin B1 down to 6 μg kg(-1) and 1.5 μg kg(-1), respectively, thus fulfilling the standards imposed by the legislation of European Union. Maize flour samples spiked with both analytes were subjected to multiplex analysis obtaining recoveries ranging from 80 to 115% and the coefficient of variation below 20%. Finally, analysis of naturally contaminated maize samples resulted in a good agreement between CL-LFIA and a validated confirmatory HPLC-UV and commercial ELISA kit, obtaining recoveries in the range 88-120%. The proposed CL-LFIA protocol is rapid, inexpensive, easy-to-use, and fit for the purpose of rapid screening of mycotoxins in maize flour. PMID:25374970

  3. Cytotoxic assessment of the regulated, co-existing mycotoxins aflatoxin B1, fumonisin B1 and ochratoxin, in single, binary and tertiary mixtures.

    Science.gov (United States)

    Clarke, Rachel; Connolly, Lisa; Frizzell, Caroline; Elliott, Christopher T

    2014-11-01

    Aflatoxin B1 (AFB1), ochratoxin A (OTA) and fumonisin B1 (FB1) are contaminants which have been shown to regularly co-occur in a range of foods. However, only a small number of studies have evaluated the interactive effect of binary and tertiary mycotoxins. The present study evaluated the effects of low levels of each mycotoxin in combination at their EU regulatory limits. Toxic effect with respect to cell viability was measured by MTT and neutral red assays, assessing mitochondria and lysosome integrities respectively. Individual toxicity showed that OTA (10 μg/ml) was the most cytotoxic mycotoxin in all three cell lines studied (caco-2, MDBK and raw 264.7). Binary combinations were cytotoxic to the MDBK cell line in the order [OTA/FB1] > [AFB1/FB1] > [AFB1/OTA], whilst all effects observed were classified as being additive. Tertiary combinations of AFB1, FB1 and OTA at the EU regulatory limits were tested and not found to exhibit measurable cytotoxicity in MDBK, caco-2 or raw 264.7 cells. However by increasing these concentrations above the legal limits to OTA (3 μg/ml), FB1 (8 μg/ml) and AFB1 (1.28 μg/ml), cytotoxicity was observed with up to 26% reduction in cell viability and synergistic effects were evident with regard to mitochondrial integrity.

  4. Mycobiota and Aflatoxin B1 in Feed for Farmed Sea Bass (Dicentrarchus labrax

    Directory of Open Access Journals (Sweden)

    Fernando Manuel d´Almeida Bernardo

    2011-02-01

    Full Text Available The safety characteristics of feed used in fish and crustacean aquaculture systems are an essential tool to assure the productivity of those animal exploitations. Safety of feed may be affected by different hazards, including biological and chemical groups. The aim of this preliminary study was to evaluate fungi contamination and the presence of aflatoxins in 87 samples of feed for sea bass, collected in Portugal. Molds were found in 35 samples (40.2% in levels ranging from 1 to 3.3 log10 CFU∙g−1. Six genera of molds were found. Aspergillus flavus was the most frequent, found in all positive samples, with a range from 2 to 3.2 log10 CFU∙g−1. Aspergillus niger was found in 34 samples (39.1%, ranging from 1 to 2.7 log10 CFU∙g−1. Aspergillus glaucus was found in 26 samples (29.9% with levels between 1 and 2.4 log10 CFU∙g−1. Penicillium spp. and Cladosporium spp. were both found in 25 samples (28.7%. Fusarium spp. was found in 22 samples (25.3%, ranging from 1 to 2.3 log10 CFU∙g−1. All feed samples were screened for aflatoxins using a HPLC technique, with a detection limit of 1.0 μg∙kg−1. All samples were aflatoxin negative.

  5. Kaempferol ameliorates aflatoxin B1 (AFB1) induced hepatocellular carcinoma through modifying metabolizing enzymes, membrane bound ATPases and mitochondrial TCA cycle enzymes

    Institute of Scientific and Technical Information of China (English)

    Kulanthaivel Langeswaran; Rajendran Revathy; Subbaraj Gowtham Kumar; Shanmugam Vijayaprakash

    2012-01-01

    Objective: The present study was aimed to scrutinize the anticancer consequence of kaempferol against aflatoxin B1 induced hepatocarcinogenesis. Epidemiological studies of the incidence of liver cancer in the population, where dietary aflatoxin exposure is high, have provided much circumstantial evidence for the development of aflatoxin B1 induced primary liver cancer in humans. Methods:In the present investigation, aflatoxin B1 (2 mg/kg body weight i.p) was used as a hepatocarcinogen to induce hepatocellular carcinoma in experimental animals. Results: In the present analysis, on treatment with bioflavonoid kaempferol (100 mg/kg body weight p.o) the nucleic acids levels were brought back to normal and also the altered levels of biological enzymes such as membrane bound ATPase, carbohydrate metabolizing enzymes and mitochondrial TCA cycle enzymes levels (P<0.01).Conclusions:Membrane bound ATPase, carbohydrate metabolizing enzymes and mitochondrial TCA cycle enzymes were modulated by kaempferol evaluated on aflatoxin B1 induced primary liver carcinogenesis.

  6. Production and characteristics of specialised monoclonal antibodies against aflatoxin B1%高特异性黄曲霉毒素B1单克隆抗体的制备及特性研究

    Institute of Scientific and Technical Information of China (English)

    肖智; 李培武; 张奇; 张文; 丁小霞

    2011-01-01

    Aflatoxins are important mycotoxins of contaminated peanut products. Aflatoxin B1 is the most toxic,accounting for over 70% of aflatoxin analogues. In present paper, aflatoxin B1 was transformed into hemiacetal,namely aflatoxin B2a, and then conjugated to protein as immunogen by reduction of sodium borohydride. After the fourth boost immunization, spleen cells of the immunized Balb/c mouse were fused with freshly isolated SP2/0 myeloma cells. Through selecting and identifying with enzyme -linked immunosorbent assay (ELISA), one clone 3A12 was obtained from cultured semi - solid medium. Sensitivity of antibody 3A12 reached 6.1 ± 0.025ng/mL.The cross - reactivities with other aflatoxin analogues ( aflatoxin B2, G1 and G2) were 7.8%, 20.2% and 0.6%respectively, with aflatoxin M1 less than 0.1%. It offered an important foundation for aflatoxin B1 -specific immuno- assay technologies and products.%本研究将黄曲霉毒素B1转化为半缩醛B2a,在硼氢化钠(NaBH4)还原作用下与载体蛋白偶联制备完全抗原.将制备的完全抗原免疫Balb/c小鼠,经4次免疫后取其脾脏与小鼠骨髓瘤细胞Sp2/0细胞融合,采用半固体培养基筛选后鉴定,获得杂交瘤细胞株3A12,抗体的灵敏度可达6.1±0.025ng/mL,抗体与其它黄曲霉毒素B2、G1及G2的交叉反应率依次为7.8%、20.2%及0.6%,与黄曲霉毒素M1交叉反应率小于0.1%.本研究为研发花生等农产品黄曲霉毒素B1特异性免疫分析技术及产品奠定了重要基础.

  7. Exposure of newborns to aflatoxin M1 and B1 from mothers' breast milk in Ankara, Turkey.

    Science.gov (United States)

    Gürbay, A; Sabuncuoğlu, S Atasayar; Girgin, G; Sahin, G; Yiğit, S; Yurdakök, M; Tekinalp, G

    2010-01-01

    Aflatoxins (AFs) are important risks for human health due to their widespread presence in foods and environment. However, contamination risk of breast milk with different pollutants including AFs is high in today's life conditions. Since breast milk is a major nutrient for infants, feeding of infants with safe milk is essential. Therefore, the objective of this study was to determine the levels of AF M(1) and B(1) in breast milk samples collected from 75 mothers in Ankara, Turkey. AF M(1) and B(1) levels were investigated by high performance liquid chromatography (HPLC) with a fluorescence detector following an extraction procedure. The limit of detection was found to be 5 ng/l. Both AFs were detected in diverse degrees in all breast milk samples: The level of AF M(1) were in the ranges of 60.90-299.99 ng/l, and AF B(1) were in the ranges of 94.50-4123.80 ng/l. These results pointed out the exposure of mothers and neonates to AF M(1) and B(1), and the necessity of further research on mycotoxin contamination both in foods and biological fluids as well as protection strategies. PMID:19850097

  8. Determination of aflatoxins B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil.

    Science.gov (United States)

    Bao, Lei; Trucksess, Mary W; White, Kevin D

    2010-01-01

    Edible oils are consumed directly, and used as ingredients in food, soaps, and skin products. However, oils such as olive oil, peanut oil, and sesame oil could be contaminated with aflatoxins, which are detrimental to human and animal health. A method using immunoaffinity column cleanup with RPLC separation and fluorescence detection (FLD) for determination of aflatoxins (AF) B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil was developed and validated. Test samples were extracted with methanol-water (55 + 45, v/v). After shaking and centrifuging, the lower layer was filtered, diluted with water, and filtered through glass microfiber filter paper. The filtrate was then passed through an immunoaffinity column, and the toxins were eluted with methanol. The toxins were then subjected to RPLC/FLD analysis after postcolumn UV photochemical derivatization. The accuracy and repeatability characteristics of the method were determined. Recoveries of AFB1 spiked at levels from 1.0 to 10.0 microg/kg in olive oil, peanut oil, and sesame oil ranged from 82.9 to 98.6%. RSDs ranged from 0.6 to 8.9%. HorRat values were < 0.2 for all of the matrixes tested. Recoveries of AF spiked at levels from 2.0 to 20.0 microg/kg ranged from 87.7 to 102.2%. RSDs ranged from 1.3 to 12.6%. HorRat values were < 0.4 for all of the matrixes tested. LC/MS/MS with multiple-reaction monitoring was used to confirm the identities of aflatoxins in a naturally contaminated peanut oil. PMID:20629398

  9. Inhibitory effect of eugenol on aflatoxin B1 production in Aspergillus parasiticus by downregulating the expression of major genes in the toxin biosynthetic pathway.

    Science.gov (United States)

    Jahanshiri, Zahra; Shams-Ghahfarokhi, Masoomeh; Allameh, Abdolamir; Razzaghi-Abyaneh, Mehdi

    2015-07-01

    Aflatoxin contamination of grains and agro-products is a serious food safety issue and a significant economic concern worldwide. In the present study, the effects of eugenol on Aspergillus parasiticus growth and aflatoxin production were studied in relation to the expression of some essential genes involved in aflatoxin biosynthetic pathway. The fungus was cultured in presence of serial two-fold concentrations of eugenol (15.62-500 μg mL(-1)) for 3 days at 28 °C. Mycelia dry weight was determined as an index of fungal growth, while aflatoxin production was assessed by high performance liquid chromatography. The expression of aflatoxin biosynthetic genes including ver-1, nor-1, pksA, omtA and aflR were evaluated by real-time PCR. Eugenol strongly inhibited A. parasiticus growth in the range of 19.16-95.83 % in a dose-dependent manner. Aflatoxin B1 production was also inhibited by the compound in the range of 15.07-98.0 %. The expressions of ver-1, nor-1, pksA, omtA and aflR genes were significantly suppressed by eugenol at concentrations of 62.5 and 125 μg mL(-1). These results indicate that eugenol may be considered as a good candidate to control toxigenic fungal growth and the subsequent contamination of food, feed and agricultural commodities by carcinogenic aflatoxins.

  10. Characterization of the cellular damage induced by Aflatoxin B1 in sea bream (Sparus aurata Linnaeus, 1758 hepatocytes

    Directory of Open Access Journals (Sweden)

    Giuseppe Crescenzo

    2010-01-01

    Full Text Available Gilthead sea bream (Sparus aurata L. is one of the most intensively farmed fish spe- cies in the Mediterranean, greatly studied for the relevant economic value, although its sensitivity to Aflatoxin B1 (AFB1 has to be investigated, yet. The aim of this study was to perform an in vitro evalua- tion of cytotoxic potential of AFB1 on S. aurata hepatocytes in order to grade the range of AFB1 toxicity, and the boundary between acute and long-term toxicity. Primary monolayer cultures of hepatocytes from S. aurata juveniles were treated with a wide range of concentrations from 5x103 ng/ml to 2x10 2x10-5 ng/ml of AFB1 for a different period of exposure (24, 48, 72 hours. The cytotoxic activity was characterized by MTT reduction assay. After each exposition hepatocytes were examined for morphologic alterations and apoptosis induction. AFB1 exposure significantly reduced cell viability in a dose- and time-depend- ent manner. Dose-response curves obtained after 24, 48 and 72 hrs revealed that prolonged exposure times lead to a significant increase of the toxicpotencyofAFB toxic potency of AFB AFB1. Ourresultsdemonstratethat Our results demonstrate that S. aurata hepatocytes are highly sensitive to AFB1 exposure. Such scientific findings could provide new insights to investigate the real impact of aflatoxin on marine farmed fish.

  11. In vitro ability of beer fermentation residue and yeast-based products to bind aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Fernanda Bovo

    2015-06-01

    Full Text Available This study aimed to verify the in vitro ability of beer fermentation residue (BFR containing Saccharomyces cerevisiae cells and five commercial products that differed in the viability and integrity of S. cerevisiae cells to remove aflatoxin B1 (AFB1 from a citrate-phosphate buffer solution (CPBS. BFR was collected at a microbrewery and prepared by drying and milling. The commercial yeast-based products were as follows: inactive intact yeast cells from beer alcoholic fermentation, inactive intact yeast cells from sugarcane alcoholic fermentation, hydrolyzed yeast cells, yeast cell walls and active yeast cells. Adsorption assays were performed in CPBS spiked with 1.0 μg AFB1/mL at pH 3.0 and 6.0 for a contact time of 60 min at room temperature. Analysis of AFB1 in the samples was performed by high performance liquid chromatography. AFB1 adsorption by the products ranged from 45.5% to 69.4% at pH 3.0 and from 24.0% to 63.8% at pH 6.0. The higher percentages (p 0.05 from commercial products containing inactive intact yeast cells. The results of this trial indicate that the yeast-based products tested, especially the BFR, have potential applications in animal feeds as a suitable biological method for reducing the adverse effects of aflatoxins.

  12. Distinct response of the hepatic transcriptome to Aflatoxin B1 induced hepatocellular carcinogenesis and resistance in rats

    Science.gov (United States)

    Shi, Jiejun; He, Jiangtu; Lin, Jing; Sun, Xin; Sun, Fenyong; Ou, Chao; Jiang, Cizhong

    2016-01-01

    Aflatoxin is a natural potent carcinogen and a major cause of liver cancer. However, the molecular mechanisms of hepatocellular carcinogenesis remain largely unexplored. In this study, we profiled global gene expression in liver tissues of rats that developed hepatocellular carcinoma (HCC) from aflatoxin B1 (AFB1) administration and those that were AFB1-resistant, as well as rats without AFB1 exposure as a control. AFB1 exposure resulted in extensive perturbation in gene expression with different functions in HCC and AFB1 resistance (AR) samples. The differentially expressed genes (DEGs) in HCC sample were enriched for cell proliferation, cell adhesion and vasculature development that largely contribute to carcinogenesis. Anti-apoptosis genes were up-regulated in HCC sample whereas apoptosis-induction genes were up-regulated in AR sample. AFB1 exposure also caused extensive alteration in expression level of lncRNAs. Among all the 4511 annotated lncRNAs, half of them were highly expressed only in HCC sample and up-regulated a group of protein-coding genes with cancer-related functions: apoptosis regulation, DNA repair, and cell cycle. Intriguingly, these genes were down-regulated by lncRNAs highly expressed in AR sample. Collectively, apoptosis is the critical biological process for carcinogenesis in response to AFB1 exposure through changes in expression level of both protein-coding and lncRNA genes. PMID:27545718

  13. Effects of feeding corn naturally contaminated with aflatoxin B1 and B2 on hepatic functions of broilers.

    Science.gov (United States)

    Yang, J; Bai, F; Zhang, K; Bai, S; Peng, X; Ding, X; Li, Y; Zhang, J; Zhao, L

    2012-11-01

    The purpose of this study was to evaluate the effects of feeding corn naturally contaminated with aflatoxin B(1) (AFB(1)) and aflatoxin B(2) (AFB(2)) on serum biochemical parameters, hepatic antioxidant enzyme activities, and pathological lesions of broilers. In total, 1,200 Cobb male broilers were randomly allocated into 5 treatments, with 8 replicates per treatment and 30 birds per replicate, in a 42-d experiment. The dietary treatments were as follows: control, 25, 50, 75, and 100% contaminated corn groups. Results showed that serum aspartate aminotransferase activity in the 75 and 100% contaminated groups were higher than that in the control group on d 21 (P contaminated corn increased (P contaminated corn (P contaminated groups on d 21 (P contaminated corn. In conclusion, diets containing AFB(1) and AFB(2) could induce pathological lesions in the livers, slightly change the serum biochemical parameters, and damage the hepatic antioxidant functions when the inclusion of AFB(1)- and AFB(2)-contaminated corn reached or exceeded 50%.

  14. Colloidal gold based immuno chromatographic strip for the simple and sensitive determination of aflatoxin B1 and B2 in corn and rice

    International Nuclear Information System (INIS)

    We have developed a simple and fast immuno chromatographic test strip for the simultaneous quantitation of aflatoxin B1 and aflatoxin B2 in corn and rice. The strip contains three pads (sample, conjugate, and absorbing pad) and uses the respective polyclonal antibodies immobilized on gold nanoparticles. Matrix interferences were minimized by application of fugacity theory. Clean-up of samples and pre-treatment of strip pads is not required. The visual detection limit is 0.1 ng mL−1, and the process can be completed within 5 min. Out of 113 natural samples, 16 rice and 27 corn samples (38% in total) were aflatoxin positive and the test results were confirmed by HPLC. The strip shows, however, high cross reactivity to aflatoxins G1, G2, and M1. We consider this strip to possess wide applicability because of its ease of use, sensitivity, stability, and low cost. (author)

  15. Investigation on Aflatoxin B1 Contamination in Chilli Products of Guizhou Province%贵州省辣椒制品中黄曲霉毒素B1的污染调查

    Institute of Scientific and Technical Information of China (English)

    高博; 王艳; 陈霄; 曹云恒; 焦彦朝

    2012-01-01

    为了解贵州省辣椒制品中黄曲霉毒素B1的污染情况,采用ELISA方法对2009-2010年贵州省9个地区市场销售的669份辣椒制品中黄曲霉毒素B1含量进行检测分析.结果表明,贵州省辣椒制品中总体存在黄曲霉毒素B1的轻度污染,尤其是干辣椒制品中黄曲霉毒素B1的污染情况较重.%In order to know the aflatoxin Bl contamination in chilli products of Guizhou Province, the aflatoxin Bl contents in 669 samples from market in nine regions of Guizhou Province were detected and analyzed by ELISA method in 2009 - 2010. The results showed that the chilli products in Guizhou Province, the overall of which existed light degree of aflatoxin Bl contamination, but the dried chilli products existed heavier aflatoxin Bl contamination.

  16. The disposition of 3H-aflatoxin B1 in the rainbow trout (Oncorhynchys mykiss) after oral and intravenous administration

    International Nuclear Information System (INIS)

    The disposition of tritiated aflatoxin B1 in the rainbow trout following oral and intravenous administration was studied over a period of 8 days by means of liquid scintillation counting and whole-body autoradiography. The pattern of distribution together with the quantitative measurements were fairly similar in both groups, indicating a high degree of gastrointestinal absorption. The highest concentrations of radioactivity were observed in the bile, liver, kidney, pyloric caeca, uveal tract of the eye and the olfactory rosette. Substantial amounts of radioactivity were still present in the liver, kidney, olfactory rosette and the mucosa of the pyloric caeca 8 days after administration. A major fraction of this radioactivity was not extractable with certain polar and nonpolar solvents, indicating covalently bound metabolites

  17. An enzyme-free catalytic DNA circuit for amplified detection of aflatoxin B1 using gold nanoparticles as colorimetric indicators

    Science.gov (United States)

    Chen, Junhua; Wen, Junlin; Zhuang, Li; Zhou, Shungui

    2016-05-01

    An enzyme-free biosensor for the amplified detection of aflatoxin B1 has been constructed based on a catalytic DNA circuit. Three biotinylated hairpin DNA probes (H1, H2, and H3) were designed as the assembly components to construct the sensing system (triplex H1-H2-H3 product). Cascaded signal amplification capability was obtained through toehold-mediated strand displacement reactions to open the hairpins and recycle the trigger DNA. By the use of streptavidin-functionalized gold nanoparticles as the signal indicators, the colorimetric readout can be observed by the naked eye. In the presence of a target, the individual nanoparticles (red) aggregate into a cross-linked network of nanoparticles (blue) via biotin-streptavidin coupling. The colorimetric assay is ultrasensitive, enabling the visual detection of trace levels of aflatoxin B1 (AFB1) as low as 10 pM without instrumentation. The calculated limit of detection (LOD) is 2 pM in terms of 3 times standard deviation over the blank response. The sensor is robust and works even when challenged with complex sample matrices such as rice samples. Our sensing platform is simple and convenient in operation, requiring only the mixing of several solutions at room temperature to achieve visible and intuitive results, and holds great promise for the point-of-use monitoring of AFB1 in environmental and food samples.An enzyme-free biosensor for the amplified detection of aflatoxin B1 has been constructed based on a catalytic DNA circuit. Three biotinylated hairpin DNA probes (H1, H2, and H3) were designed as the assembly components to construct the sensing system (triplex H1-H2-H3 product). Cascaded signal amplification capability was obtained through toehold-mediated strand displacement reactions to open the hairpins and recycle the trigger DNA. By the use of streptavidin-functionalized gold nanoparticles as the signal indicators, the colorimetric readout can be observed by the naked eye. In the presence of a target, the

  18. Carry-over of aflatoxin B1 to aflatoxin M1 in high yielding Israeli cows in mid- and late-lactation.

    Science.gov (United States)

    Britzi, Malka; Friedman, Shmulik; Miron, Joshua; Solomon, Ran; Cuneah, Olga; Shimshoni, Jakob A; Soback, Stefan; Ashkenazi, Rina; Armer, Sima; Shlosberg, Alan

    2013-01-16

    The potent hepatotoxin and carcinogen aflatoxin B1 (AFB1) is a common mycotoxin contaminant of grains used in animal feeds. Aflatoxin M1 (AFM1) is the major metabolite of AFB1 in mammals, being partially excreted into milk, and is a possible human carcinogen. The maximum permitted concentration of AFM1 in cows' milk is 0.05 μg/kg in Israel and the European Union. Since milk yield and the carry-over of AFB1 in the feed to AFM1 in the milk are highly correlated, it was considered important to determine the AFM1 carry-over in Israeli-Holstein dairy cows, distinguished by world record high milk production. Twelve such cows were used to determine AFM1 carry-over following daily oral administration of feed containing ~86 μg AFB1 for 7 days. The mean carry-over rate at steady-state (Days 3-7) was 5.8% and 2.5% in mid-lactation and late-lactation groups, respectively. The carry-over appears to increase exponentially with milk yield and could be described by the equation: carry-over% = 0.5154 e(0.0521 × milk yield), with r(2) = 0.6224. If these data truly reflect the carry-over in the national Israeli dairy herd, the maximum level of AFB1 in feed should not exceed 1.4 μg/kg, a value 3.6 times lower than the maximum residue level currently applied in Israel.

  19. Sulforaphane induces glutathione S-transferase isozymes which detoxify aflatoxin B(1)-8,9-epoxide in AML 12 cells.

    Science.gov (United States)

    Gao, Shang Shang; Chen, Xiao Yan; Zhu, Ri Zhe; Choi, Byung-Min; Kim, Bok-Ryang

    2010-01-01

    The aflatoxin B(1)-8,9-epoxide (AFBO) is hepatocarcinogenic intermediate of aflatoxin B(1) (AFB(1)) and is detoxified by glutathione S-transferases (GSTs). In this study, we investigated whether sulforaphane (SFN) could increase the rate of conjugation between AFBO and glutathione (GSH) as well as which of the GST isozymes were involved in the conjugation reaction. The conjugation potential was inhibited dose dependently with curcumin, an inhibitor of GSTs. SFN induced the expression of GST A3, GST A4, GST M1, GST P1, and GST T1 in alpha mouse line (AML) 12 cells. The cells treated with SFN (10 microM) for 12 h showed a 35-fold increase in conjugation potential of AFBO with GSH compared with the vehicle-treated cell. The conjugation potential was blocked partially by transfection of cells with siRNAs against each of the GST isozymes. The activity of GST A3 had the strongest effect on the conjugation potential. SFN treatment also increased total GST activity detected with 1-chloro-2,4-dinitrobenzene (CDNB) up to 4.3-fold. The induction fold was much lower than that detected with AFBO. These results suggest that the chemopreventive effect of SFN on the decomposition of AFBO is related to the upregulation of several GST isozymes genes. The increase of GST activity by SFN was extremely specific toward the conjugation reaction of AFBO compared with CDNB. Therefore, this system for detecting GST activity seems to be an excellent method for screening chemopreventive compounds toward AFB(1) toxicity. PMID:20818711

  20. Structure elucidation and toxicity analyses of the radiolytic products of aflatoxin B1 in methanol-water solution

    International Nuclear Information System (INIS)

    Highlights: → Radiolytic products of aflatoxin B1 were produced under gamma irradiation. → Seven key radiolytic products were structure-elucidated. → Free-radical species in radiolytic solution resulted in the formation of products. → Based on the structure-activity relationship analysis, the toxicity of radiolytic products was significantly reduced compared with that of AFB1. → The addition reaction on furan ring double bond was the reason for the reduced toxicity. - Abstract: The identification of the radiolytic products of mycotoxins is a key issue in the feasibility study of gamma ray radiation detoxification. Methanol-water solution (60:40, v/v) spiked with aflatoxin B1 (AFB1; 20 mg L-1) was irradiated with Co60 gamma ray to generate radiolytic products. Liquid chromatography-quadruple time-of-flight mass spectrometry was applied to identify the radiolytic products of AFB1. Accurate mass and proposed molecular formulas with a high-matching property of more than 20 radiolytic products were obtained. Seven key radiolytic products were proposed based on the molecular formulas and tandem mass spectrometry spectra. The analyses of toxicity and formation pathways were proposed based on the structure of the radiolytic products. The addition reaction caused by the free-radical species in the methanol-water solution resulted in the formation of most radiolytic products. Based on the structure-activity relationship analysis, the toxicity of radiolytic products was significantly reduced compared with that of AFB1 because of the addition reaction that occurred on the double bond in the terminal furan ring. For this reason, gamma irradiation is deemed an effective tool for the detoxification of AFB1.

  1. Effect of high sucrose diet on liver enzyme content and activity and aflatoxin B1-induced mutagenesis.

    Science.gov (United States)

    Peters, Leandra P; Teel, Robert W

    2003-01-01

    Aflatoxin B1 (AFB1) is a fungal toxin and contaminant that has been implicated in human liver carcinogenesis. In this study we evaluated the effect of a 65% of total calories from sucrose diet (HSD) for 90 days on hepatic cytochrome P450 (CYP450) and glutathione-S-transferase (GST) activity compared to rats maintained on standard lab chow (0% sucrose). There was a statistically significant increase in the number of S. typhimurium His+ revertants (p < 0.001) generated from the incubation of AFB1 with hepatic microsomes from rats fed a HSD. The HSD did not affect the total microsomal CYP450 content nor content of CYP450 1A2, 2B1, 2 isoforms which activate AFB1. Alkoxyresorufin O-dealkylase activity (MROD, PROD) of microsomes from animals fed HSD was decreased by 73% and 49%, respectively. MROD activity is linked to CYP 1A2 activity while PROD is linked to CYP 2B1,2 activity. Although the amount of CYP 3A was significantly decreased in rats fed a HSD, its activity, determined by the presence of the fluorometric metabolite 7-hydroxyquinidine, was unchanged. GST activity was significantly lower in the rats fed HSD.

  2. Determination of Aflatoxin B1 in Oil Plants by LC -MS/MS%LC—MS/MS测定植物油脂中黄曲霉毒素B1

    Institute of Scientific and Technical Information of China (English)

    吕飞; 朱事康; 余优军; 张奇华; 沓世远; 周宇

    2012-01-01

    An analytical method for the determination of aflatoxin B1 content in oil plants by LC - MS/MS is employed. Aflatoxin B1 testing generally uses HPLC with fluorescence detector, before column derivative, or pillars derivative, the operation is tedious, low sensi- tivity. The method adopts LC -MS/MS detection, methanol water extraction,the aflatoxin B1 immune affinity column cleansing,metha- nol capacity, LC -MS/MS detection. This method is more simple, rapid, sensitive and accurate.%建立LC—MS/MS检测植物油脂中黄曲霉毒素B1的含量。黄曲霉毒素B1的检测通常情况下都是用液相色谱检测法带荧光检测器,柱前衍生,或是柱后衍生,操作比较繁琐,灵敏度低。本法采用Lc—MS/MS检测,甲醇水提取后,经黄曲霉毒素B1免疫亲和柱,甲醇定容,LC—MS/MS检测。此法操作简便、快速、灵敏、准确。

  3. 一种新的检测黄曲霉毒素B1的酶生物传感器的制作%The Assembly of a Novel Enzyme Biosensor for Aflatoxin B1 Detection

    Institute of Scientific and Technical Information of China (English)

    刘大岭; 沈奕; 张静; 姚冬生

    2008-01-01

    A novel biosensor for aflatoxin B1 detecting has been reported. The biosensor electrode for AFB1 detecting was assembled by immobilized aflatoxin-oxidoreductase using open-ended multi-walled carbon nanotubes as matrix. Its linear range was between 0.16μM and 3.2μM. And if the specific anti-aflatoxin B1 antibody and aflatoxin oxidoreductase were both immobilized on the electrode with Multi-Walled carbon nanotubes, the detection limit of the modified electrode could be 16 nM with a 10 times improved sensitivity. The aflatoxin enzyme biosensor assembled this way strode one step forward its practical application.%报道了一种新的检测黄曲霉毒素B1的生物传感器,该传感器以开管的多壁纳米碳管固定化黄曲霉毒素氧化还原酶制作传感电极检测黄曲霉毒素B1,其线性范围达到0.16μM-3.2μM,当把特异性的黄曲霉毒素B1抗体与黄曲霉毒素氧化还原酶通过多壁纳米碳管共固定化制作修饰电极,传感器的检测限提高到16nM,灵敏度提高了10倍.用这种方法制作黄曲霉毒素酶生物传感器,使黄曲霉毒素酶生物传感器向实用化迈进了一步.

  4. Aflatoxin

    Science.gov (United States)

    ... be found in the following foods: Peanuts and peanut butter Tree nuts such as pecans Corn Wheat Oil ... tests foods that may contain aflatoxin. Peanuts and peanut butter are some of the most rigorously tested products ...

  5. The effect of NovaSil dietary supplementation on the growth and health performance of Nile tilapia (Oreochromis niloticus) fed aflatoxin-B1 contaminated feed

    Science.gov (United States)

    The objective of this study was to evaluate the ability of NovaSil (NS) clay to sorb and mitigate the toxic effects of aflatoxin B1 (AFB1) in Nile tilapia (Oreochromis niloticus). Growth performance, specific innate immunological function, intestinal microbial community, and histology were evaluate...

  6. Onderzoek naar de bruikbaarheid van twee enzym immuno assay spottests, als screeningsmethode voor de bepaling van aflatoxine B1 in samengestelde mengvoeders

    NARCIS (Netherlands)

    Trijp, van J.M.P.; Tuinstra, L.G.M.Th.

    1989-01-01

    Twee enzym immunoassay spottest zijn vergeleken met een HPLC methode voor het bepalen van aflatoxine B1. De opzet was na te gaan in hoeverre de spottests gebruikt kunnen worden als screeningsmethode van samengestelde mengvoeders bij een tolerantieniveau van10ug/kg. De testen zijn daarbij bekeken op

  7. Simultaneous determination of aflatoxin B1 and M1 in milk, fresh milk and milk powder by LC-MS/MS utilising online turbulent flow chromatography.

    Science.gov (United States)

    Fan, Sufang; Li, Qiang; Sun, Lei; Du, Yanshan; Xia, Jing; Zhang, Yan

    2015-01-01

    A novel, fully automated method based on dual-column switching using online turbulent flow chromatography followed by LC-MS/MS was developed for the determination of aflatoxin B1 and M1 in milk, fresh milk and milk powder samples. After ultrasound-assisted extraction, samples were directly injected into the chromatographic system and the analytes were concentrated on the clean-up loading column. Through purge switch, analytes were transferred to the analytical column for subsequent detection by mass spectrometry. Different types of TurboFlow(TM) columns, transfer flow rates and transfer times were optimised. Method limits of detection obtained for AFB1 and AFM1 were 0.05 μg kg(-1), and limits of quantification were 0.1 μg kg(-1). Recoveries of aflatoxin B1 and M1 were in range of 81.1-102.1% for all samples. Matrix effects of aflatoxin B1 and M1 were in range of 63.1-94.3%. The developed method was successfully used for the analysis of aflatoxin B1 and M1 in real samples. PMID:25952817

  8. Identification of early target genes of aflatoxin B1 in human hepatocytes, inter-individual variability and comparison with other genotoxic compounds

    Energy Technology Data Exchange (ETDEWEB)

    Josse, Rozenn; Dumont, Julie; Fautrel, Alain; Robin, Marie-Anne; Guillouzo, André, E-mail: andre.guillouzo@univ-rennes1.fr

    2012-01-15

    Gene expression profiling has recently emerged as a promising approach to identify early target genes and discriminate genotoxic carcinogens from non-genotoxic carcinogens and non-carcinogens. However, early gene changes induced by genotoxic compounds in human liver remain largely unknown. Primary human hepatocytes and differentiated HepaRG cells were exposed to aflatoxin B1 (AFB1) that induces DNA damage following enzyme-mediated bioactivation. Gene expression profile changes induced by a 24 h exposure of these hepatocyte models to 0.05 and 0.25 μM AFB1 were analyzed by using oligonucleotide pangenomic microarrays. The main altered signaling pathway was the p53 pathway and related functions such as cell cycle, apoptosis and DNA repair. Direct involvement of the p53 protein in response to AFB1 was verified by using siRNA directed against p53. Among the 83 well-annotated genes commonly modulated in two pools of three human hepatocyte populations and HepaRG cells, several genes were identified as altered by AFB1 for the first time. In addition, a subset of 10 AFB1-altered genes, selected upon basis of their function or tumor suppressor role, was tested in four human hepatocyte populations and in response to other chemicals. Although they exhibited large variable inter-donor fold-changes, several of these genes, particularly FHIT, BCAS3 and SMYD3, were found to be altered by various direct and other indirect genotoxic compounds and unaffected by non-genotoxic compounds. Overall, this comprehensive analysis of early gene expression changes induced by AFB1 in human hepatocytes identified a gene subset that included several genes representing potential biomarkers of genotoxic compounds. -- Highlights: ► Gene expression profile changes induced by aflatoxin B1 in human hepatocytes. ► AFB1 modulates various genes including tumor suppressor genes and proto-oncogenes. ► Important inter-individual variations in the response to AFB1. ► Some genes also altered by other

  9. Comparison of Aflatoxin B1 production by Aspergillus flavus and Aspergillus parasiticus under various conditions of temperature, light and pH

    Directory of Open Access Journals (Sweden)

    O Fani makk

    2013-07-01

    Full Text Available Abstract Background & aim: Aflatoxins are a large group of mycotoxins. The aim of the present study was the comparison of Aflatoxin B1 (AFB1 production by Aspergillus flavus (IR 111 and Aspergillus parasiticus (NRRL 2999 under various conditions of temperature, light, and pH. Methods: In this experimental study, twenty-four flasks were assigned for incubation of each of the fungi A. Flavus and parasiticus at 18, 24, 32 °C. Both flasks were maintained under conditions of light and darkness. The rate of (AFB1 produced by each groups, was measured by thin layer chromatography. Data were analyzed using descriptive statistical analysis (SPSS, version 16. Results: The lowest yield of (AFB1 produced by A. Flavus and parasiticus belonged to 32 °C and pH 6.5, respectively. On the other hand, the highest yield of toxin was observed at 24 °C and pH 6. The lighting effects were considerable. According to the studies on the adverse effects of light and the fermentation process, aflatoxin production increased in dark conditions. Conclusion: The results of study showed that A. Parasiticus (NRRL 2999 produced more aflatoxin than A. Flavus (IR 111. Key words: Aflatoxin B1, Aspergillus flavus, Aspergillus parasiticus

  10. Effects of chlorophyll and chlorophyllin on low-dose aflatoxin B1 pharmacokinetics in human volunteers: A pilot study

    Energy Technology Data Exchange (ETDEWEB)

    Jubert, C; Mata, J; Bench, G; Dashwood, R; Pereira, C; Tracewell, W; Turteltaub, K; Williams, D; Bailey, G

    2009-04-20

    Chlorophyll (Chla) and chlorophyllin (CHL) were shown previously to reduce carcinogen bioavailability, biomarker damage, and tumorigenicity in trout and rats. These findings were partially extended to humans (Proc Natl Acad Sci USA 98, 14601-14606 (2001)), where CHL reduced excretion of aflatoxin B{sub 1} (AFB{sub 1})-DNA repair products in Chinese unavoidably exposed to dietary AFB{sub 1}. However, neither AFB{sub 1} pharmacokinetics nor Chla effects were examined. We conducted a small unblinded crossover study to establish AFB{sub 1} pharmacokinetic parameters in human volunteers, and to explore possible effects of CHL or Chla co-treatment on those parameters. For protocol 1, fasted subjects received an IRB-approved dose of 14C-AFB{sub 1} (30 ng, 5 nCi) by capsule with 100 ml water, followed by normal eating and drinking after hr 2. Blood and cumulative urine samples were collected over 72 hr, and {sup 14}C-AFB{sub 1} equivalents were determined by Accelerator Mass Spectrometry. Protocols 2 and 3 were similar except capsules also contained 150 mg of purified Chla, or CHL, respectively. All protocols were repeated 3 times for each of three volunteers. The study revealed rapid human AFB{sub 1} uptake (plasma ka 5.05 {+-} 1.10 hr-1, Tmax 1.0 hr) and urinary elimination (95% complete by 24 hr) kinetics. Chla and CHL treatment each significantly impeded AFB{sub 1} absorption and reduced Cmax and AUC's (plasma and urine) in one or more subjects. These initial results provide AFB{sub 1} pharmacokinetic parameters previously unavailable for humans, and suggest that Chla or CHL co-consumption may limit the bioavailability of ingested aflatoxin in humans, as they do in animal models.

  11. Immunotoxicity of aflatoxin B1: impairment of the cell-mediated response to vaccine antigen and modulation of cytokine expression.

    Science.gov (United States)

    Meissonnier, Guylaine M; Pinton, Philippe; Laffitte, Joëlle; Cossalter, Anne-Marie; Gong, Yun Yun; Wild, Christopher P; Bertin, Gérard; Galtier, Pierre; Oswald, Isabelle P

    2008-09-01

    Aflatoxin B1 (AFB1), a mycotoxin produced by Aspergillus flavus or A. parasiticus, is a frequent contaminant of food and feed. This toxin is hepatotoxic and immunotoxic. The present study analyzed in pigs the influence of AFB1 on humoral and cellular responses, and investigated whether the immunomodulation observed is produced through interference with cytokine expression. For 28 days, pigs were fed a control diet or a diet contaminated with 385, 867 or 1807 microg pure AFB1/kg feed. At days 4 and 15, pigs were vaccinated with ovalbumin. AFB1 exposure, confirmed by an observed dose-response in blood aflatoxin-albumin adduct, had no major effect on humoral immunity as measured by plasma concentrations of total IgA, IgG and IgM and of anti-ovalbumin IgG. Toxin exposure did not impair the mitogenic response of lymphocytes but delayed and decreased their specific proliferation in response to the vaccine antigen, suggesting impaired lymphocyte activation in pigs exposed to AFB1. The expression level of pro-inflammatory (TNF-alpha, IL-1beta, IL-6, IFN-gamma) and regulatory (IL-10) cytokines was assessed by real-time PCR in spleen. A significant up-regulation of all 5 cytokines was observed in spleen from pigs exposed to the highest dose of AFB1. In pigs exposed to the medium dose, IL-6 expression was increased and a trend towards increased IFN-gamma and IL-10 was observed. In addition we demonstrate that IL-6 impaired in vitro the antigenic- but not the mitogenic-induced proliferation of lymphocytes from control pigs vaccinated with ovalbumin. These results indicate that AFB1 dietary exposure decreases cell-mediated immunity while inducing an inflammatory response. These impairments in the immune response could participate in failure of vaccination protocols and increased susceptibility to infections described in pigs exposed to AFB1.

  12. Immunotoxicity of aflatoxin B1: Impairment of the cell-mediated response to vaccine antigen and modulation of cytokine expression

    International Nuclear Information System (INIS)

    Aflatoxin B1 (AFB1), a mycotoxin produced by Aspergillus flavus or A. parasiticus, is a frequent contaminant of food and feed. This toxin is hepatotoxic and immunotoxic. The present study analyzed in pigs the influence of AFB1 on humoral and cellular responses, and investigated whether the immunomodulation observed is produced through interference with cytokine expression. For 28 days, pigs were fed a control diet or a diet contaminated with 385, 867 or 1807 μg pure AFB1/kg feed. At days 4 and 15, pigs were vaccinated with ovalbumin. AFB1 exposure, confirmed by an observed dose-response in blood aflatoxin-albumin adduct, had no major effect on humoral immunity as measured by plasma concentrations of total IgA, IgG and IgM and of anti-ovalbumin IgG. Toxin exposure did not impair the mitogenic response of lymphocytes but delayed and decreased their specific proliferation in response to the vaccine antigen, suggesting impaired lymphocyte activation in pigs exposed to AFB1. The expression level of pro-inflammatory (TNF-α, IL-1β, IL-6, IFN-γ) and regulatory (IL-10) cytokines was assessed by real-time PCR in spleen. A significant up-regulation of all 5 cytokines was observed in spleen from pigs exposed to the highest dose of AFB1. In pigs exposed to the medium dose, IL-6 expression was increased and a trend towards increased IFN-γ and IL-10 was observed. In addition we demonstrate that IL-6 impaired in vitro the antigenic- but not the mitogenic-induced proliferation of lymphocytes from control pigs vaccinated with ovalbumin. These results indicate that AFB1 dietary exposure decreases cell-mediated immunity while inducing an inflammatory response. These impairments in the immune response could participate in failure of vaccination protocols and increased susceptibility to infections described in pigs exposed to AFB1

  13. Structural Elucidation and Toxicity Assessment of Degraded Products of Aflatoxin B1 and B2 by Aqueous Extracts of Trachyspermum ammi

    Science.gov (United States)

    Iram, Wajiha; Anjum, Tehmina; Iqbal, Mazhar; Ghaffar, Abdul; Abbas, Mateen

    2016-01-01

    In this study aqueous extract of seeds and leaves of Trachyspermum ammi were evaluated for their ability to detoxify aflatoxin B1 and B2 (AFB1; 100 μg L−1 and AFB2; 50 μg L−1) by in vitro and in vivo assays. Results indicated that T. ammi seeds extract was found to be significant (P Brine shrimps bioassay further confirmed the low toxicity of degraded products, showing that T. ammi seeds extract can be used as an effective tool for the detoxification of aflatoxins. PMID:27064492

  14. Structural Analysis and Biological Toxicity of Aflatoxins B1 and B2 Degradation Products Following Detoxification by Ocimum basilicum and Cassia fistula Aqueous Extracts

    Science.gov (United States)

    Iram, Wajiha; Anjum, Tehmina; Iqbal, Mazhar; Ghaffar, Abdul; Abbas, Mateen; Khan, Abdul Muqeet

    2016-01-01

    This study showed the comparison between Ocimum basilicum and Cassia fistula (leaves and branch) aqueous extracts for their ability to detoxify of aflatoxins B1 and B2 (AFB1; 100 μg L-1 and AFB2; 50 μg L-1) by In Vitro assays and decontamination studies. Results indicated that O. basilicum leaves extract was found to be highly significant (P Brine shrimps bioassay further confirmed the low toxicity of degraded products, showing that O. basilicum leaves extract can be used as an effective tool for the detoxification of aflatoxins. PMID:27471501

  15. Impact of aflatoxin B1 on hypothalamic neuropeptides regulating feeding behavior.

    Science.gov (United States)

    Trebak, Fatima; Alaoui, Abdelilah; Alexandre, David; El Ouezzani, Seloua; Anouar, Youssef; Chartrel, Nicolas; Magoul, Rabia

    2015-07-01

    The presence of mycotoxins in food is a major problem of public health as they produce immunosuppressive, hepatotoxic and neurotoxic effects. Mycotoxins also induce mutagenic and carcinogenic effects after long exposure. Among mycotoxins that contaminate food are aflatoxins (AF) such as AFB1, which is the most powerful natural carcinogen. The AF poisoning results in symptoms of depression, anorexia, diarrhea, jaundice or anemia that can lead to death, but very few studies have explored the impact of AF on neuroendocrine regulations. To better understand the neurotoxic effects of AF related to anorexia, we explored in rat the impact of AFB1 on the major hypothalamic neuropeptides regulating feeding behavior, either orexigenic (NPY, Orexin, AgRP, MCH) or anorexigenic (α-MSH, CART, TRH). We also studied the effect of AFB1 on a novel neuropeptide, the secretogranin II (SgII)-derived peptide EM66, which has recently been linked to the control of food intake. For this, adult male rats were orally treated twice a week for 5 weeks with a low dose (150 μg/kg) or a high dose (300 μg/kg) of AFB1 dissolved in corn oil. Repeated exposure to AFB1 resulted in reduced body weight gain, which was highly significant for the high dose of AF. Immunocytochemical and quantitative PCR experiments revealed a dose-related decrease in the expression of all the hypothalamic neuropeptides studied in response to AFB1. Such orexigenic and anorexigenic alterations may underlie appetite disorders as they are correlated to a dose-dependent decrease in body weight gain of treated rats as compared to controls. We also found a decrease in the number of EM66-containing neurons in the arcuate nucleus of AFB1-treated animals, which was associated with a lower expression of its precursor SgII. These findings show for the first time that repeated consumption of AFB1 disrupts the hypothalamic regulation of neuropeptides involved in feeding behavior, which may contribute to the lower body weight gain

  16. Investigation of the Properties of Covalent Immobilized Anti-aflatoxin B1 Antibody on Membranes from Copolymer of Polyacrylamide-polyacrylonitrile

    Directory of Open Access Journals (Sweden)

    Lyubov Yotova

    2010-12-01

    Full Text Available Aflatoxins are toxic secondary metabolites produced by a number of different fungi (Aspergillus flavus, Aspergillus parasiticus, and can be present in a wide range of food and feed commodities. The most used methods for analysis of aflatoxins are thin-layer chromatography, high-performance liquid chromatography (HPLC, electrochemical immunoanalysis and microtitre plate enzyme-linked immunosorbent assay (ELISA. Membranes from copolymer of polyacrylamide-polyacrylonitrile have been prepared. These membranes were used as a matrix for a covalent binding of polyclonal anti-aflatoxin B1 antibody. ELISA was carried out with these membranes to prove successful immobilization of the antibody. It was done comparative analysis with ELISA between standard microtiter plate and our membranes with infected peanuts.

  17. [Biological contamination by micromycetes in dried Boletus edulis: research of aflatoxin B1, B2 G1, G2 and ochratoxin A].

    Science.gov (United States)

    Lorini, C; Rossetti, F; Palazzoni, S; Comodo, N; Bonaccorsi, G

    2008-01-01

    Aim of this survey is to identify those filamentous fungi which parasite Boletus edulis and its group and check the potential presence of secondary metabolites, specifically aflatoxin B1, total aflatoxins and ochratoxin A, in order to assess the risk to consumers' health. Forty samples of dried Boletus edulis, collected by two food industries which distribute the product in many Italian regions, have been analysed. The sampling plan has been conducted from November 2005 to March 2006, collecting 50 g from each commercial category of dried Boletus edulis available in the factory at the time of sampling. All the samples have been tested by visual macroscopic and stereoscopic assays; for some samples--those referred to commercial category presumably at higher risk--we have performed cultural assays as well, typization of isolated micromycetes, extraction and quantification of aflatoxins and ochratoxin A. Mycotoxin detection has been made by HPLC, using the UNI EN 14123 and UNI EN 14132 standard methods, respectively applied to aflatoxins determination in peanuts, pistachios, figs and paprika and to ochratoxin A in barley and coffee. Non pathogenic micromycetes, common in food products, have been frequently observed in cultural assays, while Aspergillus flavus and Aspergillus niger have been found in some samples. However the concentration of aflatoxins was always under the quantification limit. The survey confirm that, if the cold chain is kept throughout the process and the distribution, Boletus edulis and analogue mycetes are not a favourable substratum for the growth and the development of moulds. PMID:19238880

  18. UPLC-MS-MS法快速测定玉米中的黄曲霉毒素B1%Ultra-High Performance Liquid Chromatography Tandem Mass Spectrometry for Rapid Determination of Aflatoxin B1 in Corn

    Institute of Scientific and Technical Information of China (English)

    于成广; 黄辉

    2012-01-01

    采用高效液相色谱-串联质谱法检测玉米中的黄曲霉毒素B1.对样品前处理条件、净化和分离条件进行了优化,黄曲霉毒素B1检出限为0.02μg/kg,回收率为83.8%~95.7%.建立的方法简便、快速、灵敏度高,能够满足玉米中痕量黄曲霉毒素B1农残的高灵敏测定要求.%An analytical method for the determination of aflatoxin Bl in corn by ultra-high performance liquid chromatography tandem mass spectrometry was developed. Sample pretreatment methods and depuration conditions were optimized. The detection limit of aflatoxin Bl was 0. 02 μg/kg. The recoveries were 83.8% - 95.7%. The results indicated that the method developed is simple, rapid, sensitive, which is suit for the analysis of aflatoxin Bl pesticides in corn.

  19. Aptamer based fluorescence recovery assay for aflatoxin B1 using a quencher system composed of quantum dots and graphene oxide

    International Nuclear Information System (INIS)

    Aflatoxin B1 (AFB1), a secondary fungal metabolite of Aspergillus flavus, was employed as a model mycotoxin to establish an aptamer based assay that exploits the quenching of the fluorescence of CdTe quantum dots (Q-dots) by graphene oxide (GO). A thiolated aptamer specific for AFB1 was linked to the surface of Q-dots via ligand exchange. The fluorescence of the aptamer modified-Q-dots is strongly quenched by GO. If, however, AFB1 is added, fluorescence is restored depending on the quantity of AFB1 added. The system was evaluated both in phosphate buffer solution and in peanut oil. If performed in an aqueous system, the assay possesses good selectivity, a wide dynamic range (from 3.2 nM to 320 μM) and a low limit of detection (1.0 nM). If performed in peanut oil solution, the dynamic range is from 1.6 nM to 160 μM, and the limit of detection is 1.4 nM. In our perception, this is a simple, sensitive and selective method for the determination of AFB1 that also may be extended to the analysis of other mycotoxins. (author)

  20. Modulation of the spleen transcriptome in domestic turkey (Meleagris gallopavo) in response to aflatoxin B1 and probiotics.

    Science.gov (United States)

    Monson, Melissa S; Settlage, Robert E; Mendoza, Kristelle M; Rawal, Sumit; El-Nezami, Hani S; Coulombe, Roger A; Reed, Kent M

    2015-03-01

    Poultry are highly susceptible to the immunotoxic effects of the food-borne mycotoxin aflatoxin B1 (AFB1). Exposure impairs cell-mediated and humoral immunity, limits vaccine efficacy, and increases the incidence of costly secondary infections. We investigated the molecular mechanisms of AFB1 immunotoxicity and the ability of a Lactobacillus-based probiotic to protect against aflatoxicosis in the domestic turkey (Meleagris gallopavo). The spleen transcriptome was examined by RNA sequencing (RNA-seq) of 12 individuals representing four treatment groups. Sequences (6.9 Gb) were de novo assembled to produce over 270,000 predicted transcripts and transcript fragments. Differential expression analysis identified 982 transcripts with statistical significance in at least one comparison between treatment groups. Transcripts with known immune functions comprised 27.6 % of significant expression changes in the AFB1-exposed group. Short exposure to AFB1 suppressed innate immune transcripts, especially from antimicrobial genes, but increased the expression of transcripts from E3 ubiquitin-protein ligase CBL-B and multiple interleukin-2 response genes. Up-regulation of transcripts from lymphotactin, granzyme A, and perforin 1 could indicate either increased cytotoxic potential or activation-induced cell death in the spleen during aflatoxicosis. Supplementation with probiotics was found to ameliorate AFB1-induced expression changes for multiple transcripts from antimicrobial and IL-2-response genes. However, probiotics had an overall suppressive effect on immune-related transcripts. PMID:25597949

  1. Sulforaphane, a cancer chemopreventive agent, induces pathways associated with membrane biosynthesis in response to tissue damage by aflatoxin B1.

    Science.gov (United States)

    Techapiesancharoenkij, Nirachara; Fiala, Jeannette L A; Navasumrit, Panida; Croy, Robert G; Wogan, Gerald N; Groopman, John D; Ruchirawat, Mathuros; Essigmann, John M

    2015-01-01

    Aflatoxin B1 (AFB1) is one of the major risk factors for liver cancer globally. A recent study showed that sulforaphane (SF), a potent inducer of phase II enzymes that occurs naturally in widely consumed vegetables, effectively induces hepatic glutathione S-transferases (GSTs) and reduces levels of hepatic AFB1-DNA adducts in AFB1-exposed Sprague Dawley rats. The present study characterized the effects of SF pre-treatment on global gene expression in the livers of similarly treated male rats. Combined treatment with AFB1 and SF caused reprogramming of a network of genes involved in signal transduction and transcription. Changes in gene regulation were observable 4h after AFB1 administration in SF-pretreated animals and may reflect regeneration of cells in the wake of AFB1-induced hepatotoxicity. At 24h after AFB1 administration, significant induction of genes that play roles in cellular lipid metabolism and acetyl-CoA biosynthesis was detected in SF-pretreated AFB1-dosed rats. Induction of this group of genes may indicate a metabolic shift toward glycolysis and fatty acid synthesis to generate and maintain pools of intermediate molecules required for tissue repair, cell growth and compensatory hepatic cell proliferation. Collectively, gene expression data from this study provide insights into molecular mechanisms underlying the protective effects of SF against AFB1 hepatotoxicity and hepatocarcinogenicity, in addition to the chemopreventive activity of this compound as a GST inducer. PMID:25450479

  2. Modulation of the spleen transcriptome in domestic turkey (Meleagris gallopavo) in response to aflatoxin B1 and probiotics.

    Science.gov (United States)

    Monson, Melissa S; Settlage, Robert E; Mendoza, Kristelle M; Rawal, Sumit; El-Nezami, Hani S; Coulombe, Roger A; Reed, Kent M

    2015-03-01

    Poultry are highly susceptible to the immunotoxic effects of the food-borne mycotoxin aflatoxin B1 (AFB1). Exposure impairs cell-mediated and humoral immunity, limits vaccine efficacy, and increases the incidence of costly secondary infections. We investigated the molecular mechanisms of AFB1 immunotoxicity and the ability of a Lactobacillus-based probiotic to protect against aflatoxicosis in the domestic turkey (Meleagris gallopavo). The spleen transcriptome was examined by RNA sequencing (RNA-seq) of 12 individuals representing four treatment groups. Sequences (6.9 Gb) were de novo assembled to produce over 270,000 predicted transcripts and transcript fragments. Differential expression analysis identified 982 transcripts with statistical significance in at least one comparison between treatment groups. Transcripts with known immune functions comprised 27.6 % of significant expression changes in the AFB1-exposed group. Short exposure to AFB1 suppressed innate immune transcripts, especially from antimicrobial genes, but increased the expression of transcripts from E3 ubiquitin-protein ligase CBL-B and multiple interleukin-2 response genes. Up-regulation of transcripts from lymphotactin, granzyme A, and perforin 1 could indicate either increased cytotoxic potential or activation-induced cell death in the spleen during aflatoxicosis. Supplementation with probiotics was found to ameliorate AFB1-induced expression changes for multiple transcripts from antimicrobial and IL-2-response genes. However, probiotics had an overall suppressive effect on immune-related transcripts.

  3. Highly sensitive SERS-based immunoassay of aflatoxin B1 using silica-encapsulated hollow gold nanoparticles.

    Science.gov (United States)

    Ko, Juhui; Lee, Chankil; Choo, Jaebum

    2015-03-21

    Aflatoxin B1 (AFB1) is a well-known carcinogenic contaminant in foods. It is classified as an extremely hazardous compound because of its potential toxicity to the human nervous system. AFB1 has also been extensively used as a biochemical marker to evaluate the degree of food spoilage. In this study, a novel surface-enhanced Raman scattering (SERS)-based immunoassay platform using silica-encapsulated hollow gold nanoparticles (SEHGNs) and magnetic beads was developed for highly sensitive detection of AFB1. SEHGNs were used as highly stable SERS-encoding nano tags, and magnetic beads were used as supporting substrates for the high-density loading of immunocomplexes. Quantitative analysis of AFB1 was performed by monitoring the intensity change of the characteristic peaks of Raman reporter molecules. The limit of detection (LOD) of AFB1, determined by this SERS-based immunoassay, was determined to be 0.1 ng/mL. This method has some advantages over other analytical methods with respect to rapid analysis (less than 30 min), good selectivity, and reproducibility. The proposed method is expected to be a new analytical tool for the trace analysis of various mycotoxins.

  4. Potential Antioxidant Role of Tridham in Managing Oxidative Stress against Aflatoxin-B1-Induced Experimental Hepatocellular Carcinoma

    Directory of Open Access Journals (Sweden)

    Vijaya Ravinayagam

    2012-01-01

    Full Text Available Hepatocellular carcinoma (HCC is one of the most fatal cancers due to delayed diagnosis and lack of effective treatment options. Significant exposure to Aflatoxin B1 (AFB1, a potent hepatotoxic and hepatocarcinogenic mycotoxin, plays a major role in liver carcinogenesis through oxidative tissue damage and p53 mutation. The present study emphasizes the anticarcinogenic effect of Tridham (TD, a polyherbal traditional medicine, on AFB1-induced HCC in male Wistar rats. AFB1-administered HCC-bearing rats (Group II showed increased levels of lipid peroxides (LPOs, thiobarbituric acid substances (TBARs, and protein carbonyls (PCOs and decreased levels of enzymic and nonenzymic antioxidants when compared to control animals (Group I. Administration of TD orally (300 mg/kg body weight/day for 45 days to HCC-bearing animals (Group III significantly reduced the tissue damage accompanied by restoration of the levels of antioxidants. Histological observation confirmed the induction of tumour in Group II animals and complete regression of tumour in Group III animals. This study highlights the potent antioxidant properties of TD which contribute to its therapeutic effect in AFB1-induced HCC in rats.

  5. Preparation and characterization of an immunoaffinity column for the selective extraction of aflatoxin B1 in 13 kinds of foodstuffs.

    Science.gov (United States)

    Xie, Jie; Peng, Tao; He, Jian-Li; Shao, Yu; Fan, Chun-Lin; Chen, Ying; Jiang, Wen-Xiao; Chen, Min; Wang, Qi; Pei, Xing-Yao; Ding, Shuang-Yang; Jiang, Hai-Yang

    2015-08-15

    A rapid and reliable immunoaffinity column (IAC) clean-up based ultra performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS) method was developed for the determination of aflatoxin B1 (AFB1) in cereals, peanuts, vegetable oils and Chinese traditional food products like sufu and lobster sauce. The immunoaffinity column of AFB1 (AFB1-IAC) was prepared by coupling CNBr-activated Sepharose-4B with the anti-AFB1 monoclonal antibody. The column capacity of IAC was over 260ng/mL gel. Samples were extracted with methanol-water (60:40, v/v) and the extracts were then purified on an AFB1-IAC before UPLC-MS/MS analysis. The average recoveries of AFB1 in spiked samples at levels of 1.0, 5.0 and 10.0μg/kg ranged from 72% to 98%, with the relative standard deviations of 1.2-9.3% (n=6). The limits of qualification ranged from 0.07 to 0.23μg/kg, which were below the MRLs of AFB1 in the matrices evaluated. In this work, the developed method was suitable for the determination of trace AFB1 residues in 13 kinds of foodstuffs. PMID:26160471

  6. A simple aptamer-based fluorescent assay for the detection of Aflatoxin B1 in infant rice cereal.

    Science.gov (United States)

    Chen, Lu; Wen, Fang; Li, Ming; Guo, Xiaodong; Li, Songli; Zheng, Nan; Wang, Jiaqi

    2017-01-15

    A fluorescent assay for the rapid, sensitive and specific detection of Aflatoxin B1 (AFB1) was developed in this study. Initially, a DNA/DNA duplex was formed between a fluorescein-labeled AFB1 aptamer and its partially complementary DNA strand containing a quencher moiety, resulting in fluorescence quenching due to the close proximity of fluorophore and quencher. Upon the addition of AFB1, an aptamer/AFB1 complex was generated to release the quencher-modified DNA strand, thus recovered the fluorescence of fluorescein and enabled quantitative detection for AFB1 by monitoring fluorescence enhancement. Under optimized conditions, this assay exhibited a linear response to AFB1 in the range of 5-100ng/mL with a detection limit down to 1.6ng/mL. Trials of this assay in infant rice cereal with satisfactory recovery in the range of 93.0%-106.8%, demonstrate that the new assay could be a potential sensing platform for AFB1 determination in food. PMID:27542489

  7. Tetramethyl-6-carboxyrhodamine quenching-based aptasensing platform for aflatoxin B1: Analytical performance comparison of two aptamers.

    Science.gov (United States)

    Goud, K Yugender; Sharma, Atul; Hayat, Akhtar; Catanante, Gaëlle; Gobi, K Vengatajalabathy; Gurban, Ana Maria; Marty, Jean Louis

    2016-09-01

    In this study, a simple TAMRA (tetramethyl-6-carboxyrhodamine) quenching-based aptasensing platform was designed for the detection of aflatoxin B1 (AFB1). Here, we compared the analytical performance of two aptamer sequences: seqA and seqB. The AFB1 detection was based on the interactions of FAM (carboxyfluorescein)-labeled aptamer with TAMRA-labeled DNA complementary strand in the presence and absence of target analyte. Under optimized experimental conditions, TAMRA-labeled strand quenched the fluorescence response of FAM-labeled aptamer due to the noncovalent interaction between the two DNA strands. The binding of AFB1 induced the complex formation and weakened the interaction between FAM-labeled aptamer and TAMRA-labeled complementary strand, resulting in the fluorescence recovery. By using this principle concept, an assay was constructed for the detection of AFB1. The method exhibited good sensitivity, good selectivity with a limit of detection of 0.2 ng ml(-1), and a wide linear range from 0.25 to 32 ng ml(-1). For real sample application, the aptasensors were tested in beer and wine samples, with good recovery rates obtained for AFB1 detection. PMID:27251432

  8. Fluorescence enhancement of the aflatoxin B1 by forming inclusion complexes with some cyclodextrins and molecular modeling study

    International Nuclear Information System (INIS)

    The interaction between the aflatoxin B1 (AFB1) and three cyclodextrins, α-cyclodextrin (α-CD), β-cyclodextrin (β-CD) and heptakis-2,6-dimethyl-o-β-cyclodextrin (ome-CD), was studied by spectrofluorescence technique. It was found that the inclusion association behavior occurs for the complexes of cyclodextrins with AFB1. The fluorescence of AFB1 is generally enhanced in the complexes with cyclodextrins in aqueous solutions. The inclusion complex constants of the three types of cyclodextrins at different temperatures were evaluated from Benesi-Hildebrand plot and also by non-linear regression analysis. These cyclodextrins can only form the 1:1 (host:guest) inclusion complex in the studied temperature range of 20-50 deg. C. The enthalpy (ΔHo) and entropy (ΔSo) changes of complexation were extracted from the temperature dependency of complex formation constants (K). Temperature-dependent measurements showed that the association step is controlled by enthalpy-entropy compensation effect. The use of ome-CD generally resulted in the greatest fluorescence intensity. On the other hand, the discrepancy between the exhibited enhanced fluorescence and thermodynamic parameters (ΔGo) is proposed to be different only by the orientation of the AFB1 within the cyclodextrin cavity. To find the most favorable structure, the geometry of complex was investigated by molecular modeling approach employing the semiemperical HF-SCF calculations

  9. Simple and sensitive detection of aflatoxin B1 within five minute using a non-conventional competitive immunosensing mode.

    Science.gov (United States)

    Lin, Youxiu; Zhou, Qian; Lin, Yuping; Tang, Dianyong; Chen, Guonan; Tang, Dianping

    2015-12-15

    A novel competitive-type immunosensing strategy based on target-induced displacement reaction with antibody-functionalized mesoporous carbon (MSC) nanoparticles was designed for sensitive and rapid electrochemical detection of aflatoxin B1 (AFB1, used as a model) on the Nafion-functionalized sensing interface. Electroactive thionine molecules were initially decorated to the mesoporous carbon, and polyclonal anti-AFB1 antibody was then covalently conjugated to the nanostructures. The immunosensor was simply prepared via the electrostatic interaction between negatively charged Nafion film and positively charged anti-AFB1 antibody accompanying the nanostructures. The electrochemical signal originated from the carried thionine to mesoporous carbon. Upon target AFB1 introduction, the analyte reacted with the labeled anti-AFB1 antibody on the MSC based on specific antigen-antibody reaction and induced the dissociation of thionine-MSN nanostructures from the sensing interface, thus decreasing the cathodic current of the carried thionine molecules. Under optimal conditions, the immunosensor exhibited good electrochemical responses for determination of target AFB1 at a concentration as low as 3.0 pg mL(-1) (3.0 ppt). Importantly, the non-conventional sensing system provides a promising immunosensing strategy for rapid screening of small molecules because of its simplicity, low cost and sensitivity without the needs of sample separation and washing step.

  10. Highly sensitive SERS-based immunoassay of aflatoxin B1 using silica-encapsulated hollow gold nanoparticles.

    Science.gov (United States)

    Ko, Juhui; Lee, Chankil; Choo, Jaebum

    2015-03-21

    Aflatoxin B1 (AFB1) is a well-known carcinogenic contaminant in foods. It is classified as an extremely hazardous compound because of its potential toxicity to the human nervous system. AFB1 has also been extensively used as a biochemical marker to evaluate the degree of food spoilage. In this study, a novel surface-enhanced Raman scattering (SERS)-based immunoassay platform using silica-encapsulated hollow gold nanoparticles (SEHGNs) and magnetic beads was developed for highly sensitive detection of AFB1. SEHGNs were used as highly stable SERS-encoding nano tags, and magnetic beads were used as supporting substrates for the high-density loading of immunocomplexes. Quantitative analysis of AFB1 was performed by monitoring the intensity change of the characteristic peaks of Raman reporter molecules. The limit of detection (LOD) of AFB1, determined by this SERS-based immunoassay, was determined to be 0.1 ng/mL. This method has some advantages over other analytical methods with respect to rapid analysis (less than 30 min), good selectivity, and reproducibility. The proposed method is expected to be a new analytical tool for the trace analysis of various mycotoxins. PMID:25462866

  11. Lophirones B and C prevent aflatoxin B1-induced oxidative stress and DNA fragmentation in rat hepatocytes.

    Science.gov (United States)

    Ajiboye, Taofeek Olakunle; Yakubu, Musa Toyin; Oladiji, Adenike Temidayo

    2016-10-01

    Context Despite the reported anticarcinogenic activity of lophirones B and C, no scientific information exists for its activity in rat hepatocytes. Objective Effect of lophirones B and C on aflatoxin B1 (AFB1)-induced oxidative stress, and DNA fragmentation in rat hepatocytes was investigated. Materials and methods Wistar rat hepatocytes were incubated with lophirones B and C (1 mg/mL) or sylimarin (1 mg/mL) in the presence or absence of AFB1. For an in vivo study, rats were orally administered with lophirones B and C, and/or AFB1 (20 μg/d) for 9 weeks. Results Lophirones B and C lowered AFB1-mediated increase in nitric oxide, superoxide anion radicals, caspase-3 and fragmented DNA. Lophirones B and C attenuated AFB1-mediated decrease in superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, and reduced glutathione. Also, lophirones B and C attenuated AFB1-mediated increase in conjugated dienes, lipid hydroperoxides and malondialdehyde in rat hepatocytes. Furthermore, AFB1-mediated alterations in alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase, albumin, total bilirubin and globulin in rat serum were significantly annulled in lophirones B and C-treated rats. Conclusion This study revealed that lophirones B and C prevented AFB1-induced oxidative damage in rat hepatocytes.

  12. Incidência de fumonisina B1, aflatoxinas B1, B2, G1 e G2, ocratoxina A e zearalenona em produtos de milho Occurrence of fumonisin B1, aflatoxins B1, B2, G1, and G2, ochratoxin A and zearalenone in corn products

    Directory of Open Access Journals (Sweden)

    Luciane Mie Kawashima

    2006-09-01

    Full Text Available Levantamentos de ocorrência de micotoxinas em alimentos foram realizados nas últimas duas décadas nas regiões Sudeste e Sul do Brasil. Levantamentos em alimentos comercializados em outras regiões têm-se limitado a aflatoxinas em amendoim e castanhas do Brasil. O presente trabalho pesquisou a presença de fumonisina B1, aflatoxinas B1, B2, G1 e G2, ocratoxina A e zearalenona em 74 amostras de produtos a base de milho adquiridas no comércio da cidade de Recife, PE, durante o período de 1999 a 2001. Fumonisina B1 foi determinada por cromatografia líquida de alta eficiência com detecção por fluorescência e as demais toxinas foram determinadas por cromatografia em camada delgada. Fumonisina B1 foi encontrada em 94,6% das amostras em concentrações variando de 20 a 8600 µg/kg. Apenas 5 amostras continham aflatoxina B1 e o teor máximo encontrado foi 20 µg/kg. Duas amostras ultrapassaram o limite de 20 µg/kg para a somatória das aflatoxinas B1, B2, G1 e G2 (farinha de milho pré-cozida com 21,5 µg/kg e quirera (xerém com 23,3 µg/kg. As aflatoxinas G1 e G2, ocratoxina A e zearalenona não foram detectadas em nenhuma das amostras. Todas as amostras contaminadas com aflatoxinas também apresentaram fumonisina B1.Research concerning the presence of mycotoxin in food has been conducted in the Southwest and South regions of Brazil over the last two decades. Research in other regions has been limited to aflatoxin in peanuts and Brazil nuts. The aim of this work is to study the presence of fumonisin B1, aflatoxins B1, B2, G1, and G2, ochratoxin A and zearalenone in 74 samples of corn products acquired in shops and food markets in the city of Recife (PE from 1999 to 2001. Fumonisin B1 was determined by high performance liquid chromatography and fluorescence was detected. The other toxins were determined by thin layer chromatography. Fumonisin B1 was found in 94.6% of the samples in levels from 20 to 8600 µg/kg. Only 5 samples contained

  13. Rapid analysis of aflatoxins B1, B2, and ochratoxin A in rice samples using dispersive liquid-liquid microextraction combined with HPLC.

    Science.gov (United States)

    Lai, Xian-Wen; Sun, Dai-Li; Ruan, Chun-Qiang; Zhang, He; Liu, Cheng-Lan

    2014-01-01

    A novel, simple, and rapid method is presented for the analysis of aflatoxin B1, aflatoxin B2, and ochratoxin A in rice samples by dispersive liquid-liquid microextraction combined with LC and fluorescence detection. After extraction of the rice samples with a mixture of acetonitrile/water/acetic acid, mycotoxins were rapidly partitioned into a small volume of organic solvent (chloroform) by dispersive liquid-liquid microextraction. The three mycotoxins were simultaneously determined by LC with fluorescence detection after precolumn derivatization for aflatoxin B1 and B2. Parameters affecting both extraction and dispersive liquid-liquid microextraction procedures, including the extraction solvent, the type and volume of extractant, the volume of dispersive solvent, the addition of salt, the pH and the extraction time, were optimized. The optimized protocol provided an enrichment factor of approximately 1.25 and with detection of limits (0.06-0.5 μg/kg) below the maximum levels imposed by current regulations for aflatoxins and ochratoxin A. The mean recovery of three mycotoxins ranged from 82.9-112%, with a RSD less than 7.9% in all cases. The method was successfully applied to measure mycotoxins in commercial rice samples collected from local supermarkets in China.

  14. Development of a microwave-assisted-extraction-based method for the determination of aflatoxins B1, G1, B2, and G2 in grains and grain products.

    Science.gov (United States)

    Chen, Si; Zhang, Hong

    2013-02-01

    This article describes the use of microwave-assisted extraction (MAE) as a pretreatment technique for the determination of aflatoxins B(1), G(1), B(2), and G(2) in grains and grain products. The optimal operation parameters, including extraction solvent, temperature, and time, were identified to be acetonitrile as the extraction solvent at 80 °C with 15 min of MAE. The extracts were cleaned up using solid-phase extraction followed by derivatization with trifluoroacetic acid and were determined by liquid chromatography-fluorescence detection. A Sep-Pak cartridge was chosen over Oasis HLB and Bond Elut cartridges. By the use of aflatoxin M(1) as an internal standard, relative recoveries of the aflatoxins ranged from 90.7 to 105.7 % for corn and from 88.1 to 103.4 % for wheat, with relative standard deviations between 2.5 and 8.7 %. A total of 36 samples from local markets were analyzed, and aflatoxin B(1) was found to be the predominant toxin, with concentrations ranging from 0.42 to 3.41 μg/kg. PMID:23314480

  15. Rapid analysis of aflatoxins B1, B2, and ochratoxin A in rice samples using dispersive liquid-liquid microextraction combined with HPLC.

    Science.gov (United States)

    Lai, Xian-Wen; Sun, Dai-Li; Ruan, Chun-Qiang; Zhang, He; Liu, Cheng-Lan

    2014-01-01

    A novel, simple, and rapid method is presented for the analysis of aflatoxin B1, aflatoxin B2, and ochratoxin A in rice samples by dispersive liquid-liquid microextraction combined with LC and fluorescence detection. After extraction of the rice samples with a mixture of acetonitrile/water/acetic acid, mycotoxins were rapidly partitioned into a small volume of organic solvent (chloroform) by dispersive liquid-liquid microextraction. The three mycotoxins were simultaneously determined by LC with fluorescence detection after precolumn derivatization for aflatoxin B1 and B2. Parameters affecting both extraction and dispersive liquid-liquid microextraction procedures, including the extraction solvent, the type and volume of extractant, the volume of dispersive solvent, the addition of salt, the pH and the extraction time, were optimized. The optimized protocol provided an enrichment factor of approximately 1.25 and with detection of limits (0.06-0.5 μg/kg) below the maximum levels imposed by current regulations for aflatoxins and ochratoxin A. The mean recovery of three mycotoxins ranged from 82.9-112%, with a RSD less than 7.9% in all cases. The method was successfully applied to measure mycotoxins in commercial rice samples collected from local supermarkets in China. PMID:24243826

  16. 上海地区大米样品黄曲霉素B1含量检测%Detection of amount of Aflatoxin B1 in rice samples in Shanghai

    Institute of Scientific and Technical Information of China (English)

    叶露; 韦艳霞; 刘畅

    2011-01-01

    目的 对上海市区内出售的大米样本进行黄曲霉素B1含量的测定和分析.方法 以在上海市区内的散装粮食店、中型超市、大型超市和淘宝网店采购的苏北产大米和东北产大米作为受试样本,采用酶联免疫吸附法测定样本大米中黄曲霉素B1的含量.采用独立样本T检验分析不同产地样本大米中黄曲霉素B1含量的差异;单因素方差分析比较不同购物来源样本大米中黄曲霉素B1的含量差异.结果 60个受试样本中黄曲霉素B1含量的总体均值为1.926 ppb,其中超过国家标准样本数为2个,分别为购自散装店和中型超市的东北产大米;苏北和东北样本大米中黄曲霉素B1的含量分别为(1.221±1.055)ppb和(2.631±2.903) ppb,两者差异具有统计学意义(P<0.05).散装店、网店、中型超市和大型超市样本大米中黄曲霉素B1的含量分别为(2.900±2.457)ppb、(1.570±1.416)ppb、(1.743±2.683)ppb和(0.569±0.608) ppb,四者比较差异有统计学意义(P<0.05),进一步的多重检验表明散装店与大型超市来源样本大米中黄曲霉素B1的含量存在统计学差异(P<0.05).结论 上海市区出售的大米样本中黄曲霉素B1含量总体较低.受试样本中的黄曲霉素B1的含量,东北产大米高于苏北产大米,散装粮食店来源大米高于大型超市来源大米.在散装店和中型超市的受试样本中发现黄曲霉素B1含量超标样本.%Objective To determine and analyse the amount of Aflatoxin B, in rice samples sold in Shanghai. Methods Subei and Dongbei rice samples were obtained from groceries, medium-size markets, supermarkets and e-shops, and the amount of Aflatoxin B, in rice samples was determined by enzyme linked immunosorbent assay. The amount of Aflatoxin B, in Subei and Dongbei rice samples was compared by T test, and the amount of Aflatoxin B, in rice samples obtained through different methods was compared by one-way analysis of variance. Results

  17. An in vitro study of alkaline phosphatase sensitivity to mixture of aflatoxin B1 and fumonisin B1 in the hepatopancreas of coastal lagoon wild and farmed shrimp Litopenaeus vannamei.

    Science.gov (United States)

    Pérez-Acosta, Jesús A; Burgos-Hernandez, Armando; Velázquez-Contreras, Carlos A; Márquez-Ríos, Enrique; Torres-Arreola, Wilfrido; Arvizu-Flores, Aldo A; Ezquerra-Brauer, J Marina

    2016-08-01

    This study aimed to establish the combined effect of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) on wild Litopenaeus vannamei hepatopancreas alkaline phosphatase (AP) activity compared with that of farmed shrimp. AP activity in hepatopancreas extract was confirmed by several specific inhibitor assays. AP activity of wild shrimp was higher than that of farmed shrimp (p mycotoxins concentrations used in the study, AP from farmed L. vannamei was sensitive to the presence of both mycotoxins; however, AP is more sensitive to the combination of AFB1 + FB1 suggesting a possible synergistic or potentiating inhibitory effect. PMID:27040818

  18. Identification of early target genes of aflatoxin B1 in human hepatocytes, inter-individual variability and comparison with other genotoxic compounds.

    Science.gov (United States)

    Josse, Rozenn; Dumont, Julie; Fautrel, Alain; Robin, Marie-Anne; Guillouzo, André

    2012-01-15

    Gene expression profiling has recently emerged as a promising approach to identify early target genes and discriminate genotoxic carcinogens from non-genotoxic carcinogens and non-carcinogens. However, early gene changes induced by genotoxic compounds in human liver remain largely unknown. Primary human hepatocytes and differentiated HepaRG cells were exposed to aflatoxin B1 (AFB1) that induces DNA damage following enzyme-mediated bioactivation. Gene expression profile changes induced by a 24h exposure of these hepatocyte models to 0.05 and 0.25μM AFB1 were analyzed by using oligonucleotide pangenomic microarrays. The main altered signaling pathway was the p53 pathway and related functions such as cell cycle, apoptosis and DNA repair. Direct involvement of the p53 protein in response to AFB1 was verified by using siRNA directed against p53. Among the 83 well-annotated genes commonly modulated in two pools of three human hepatocyte populations and HepaRG cells, several genes were identified as altered by AFB1 for the first time. In addition, a subset of 10 AFB1-altered genes, selected upon basis of their function or tumor suppressor role, was tested in four human hepatocyte populations and in response to other chemicals. Although they exhibited large variable inter-donor fold-changes, several of these genes, particularly FHIT, BCAS3 and SMYD3, were found to be altered by various direct and other indirect genotoxic compounds and unaffected by non-genotoxic compounds. Overall, this comprehensive analysis of early gene expression changes induced by AFB1 in human hepatocytes identified a gene subset that included several genes representing potential biomarkers of genotoxic compounds. PMID:22100608

  19. Aflatoxin B1 Detection Using a Highly-Sensitive Molecularly-Imprinted Electrochemical Sensor Based on an Electropolymerized Metal Organic Framework

    OpenAIRE

    Mengjuan Jiang; Mohamed Braiek; Anca Florea; Amani Chrouda; Carole Farre; Anne Bonhomme; Francois Bessueille; Francis Vocanson; Aidong Zhang; Nicole Jaffrezic-Renault

    2015-01-01

    A sensitive electrochemical molecularly-imprinted sensor was developed for the detection of aflatoxin B1 (AFB1), by electropolymerization of p-aminothiophenol-functionalized gold nanoparticles in the presence of AFB1 as a template molecule. The extraction of the template leads to the formation of cavities that are able to specifically recognize and bind AFB1 through π-π interactions between AFB1 molecules and aniline moities. The performance of the developed sensor for the detection of AF...

  20. Study on Degradation Effect of Irradiation on Aflatoxin B1%辐照处理对黄曲霉毒素 B1的降解效果研究

    Institute of Scientific and Technical Information of China (English)

    王守经; 柳尧波; 胡鹏; 汝医; 王兆华; 孙宏春; 许方佐; 王志东

    2015-01-01

    利用不同剂量的高能电子束和γ射线进行辐照处理,研究了2种射线对不同状态下黄曲霉毒素B1(Aflatoxin,AFB1)的辐射降解效果和对小麦面粉粉质指标的影响。结果表明:高能电子束和γ射线辐照处理,能够降解不同状态下的 AFB1,辐照剂量越大,降解率越高,AFB1浓度越高,辐射降解率越低;相同处理剂量对溶液中黄曲霉毒素 B1的降解作用,大于对污染小麦中 AFB1的降解作用。2种辐照处理方式对面团吸水率、形成时间、稳定时间等小麦粉质指标产生了显著的影响,其中高能电子束射线的作用强度大于γ射线。%After radiating with high energy electron beam and gamma -ray at different doses,the degra-dation rates of aflatoxin B1 in methanol -water solution and contaminated wheat were studied,and the farino-graph indexes of wheat flour was determined.The results showed that the two irradiation methods could both effectively degrade aflatoxin B1 in different states.The bigger the radiation dose was,the higher the degrada-tion rate was,and the greater the aflatoxin B1 concentration was,the lower the degradation rate was.Under the same radiation dose,the degradation of aflatoxin B1 in methanol -water solution was much easier than that in contaminated wheat.Compared to the non -irradiation treatment,the two irradiation methods both signifi-cantly affected the dough water absorption rate,development time and stability time,and the effect of high en-ergy electron beam was more effective than that of gamma -ray.

  1. Organophilic treatments of bentonite increase the adsorption of aflatoxin B1 and protect stem cells against cellular damage.

    Science.gov (United States)

    Nones, Janaína; Nones, Jader; Poli, Anicleto; Trentin, Andrea Gonçalves; Riella, Humberto Gracher; Kuhnen, Nivaldo Cabral

    2016-09-01

    Bentonite clays exhibit high adsorptive capacity for contaminants, including aflatoxin B1 (AFB1), a mycotoxin responsible for causing severe toxicity in several species including pigs, poultry and man. Organophilic treatments is known to increase the adsorption capacity of bentonites, and the primary aim of this study was to evaluate the ability of Brazilian bentonite and two organic salts - benzalkonium chloride (BAC) and cetyltrimethylammonium bromide (CTAB) to adsorb AFB1. For this end, 2(2) factorial designs were used in order to analyze if BAC or CTAB was able to increase AFB1 adsorption when submitted in different temperature and concentration. Both BAC and CTAB treatment (at 30°C and 2% of salt concentration) were found to increase the adsorption of AFB1 significantly compared with untreated bentonite. After organophilic bentonite treatments with BAC or CTAB, a vibration of CH stretch (2850 and 2920cm(-1)) were detected. A frequency of the SiO stretch (1020 and 1090cm(-1)) was changed by intercalation of organic cation. Furthermore, the interlayer spacing of bentonite increases to 1.23nm (d001 reflection at 2θ=7.16) and 1.22 (d001 reflection at 2θ=7.22) after the addition of BAC and CTAB, respectively. Another aim of the study was to observe the effects of these two bentonite salts in neural crest stem cell cultures. The two materials that were created by organophilic treatments were not found to be toxic to stem cells. Furthermore the results indicate that the two materials tested may protect the neural crest stem cells against damage caused by AFB1. PMID:27281241

  2. Rapid and label-free detection of ochratoxin A and aflatoxin B1 using an optical portable instrument.

    Science.gov (United States)

    Arduini, Fabiana; Neagu, Daniela; Pagliarini, Valeria; Scognamiglio, Viviana; Leonardis, Maria Antonietta; Gatto, Emanuela; Amine, Aziz; Palleschi, Giuseppe; Moscone, Danila

    2016-04-01

    In this study, we report a novel assay for the combined on site detection of aflatoxin B1 (AFB1) and ochratoxin A (OTA), through a colorimetric biosensing system for AFB1 and a fluorimetric detection for OTA, exploiting the capability of the portable fibre optic spectrometer to perform both analyses. AFB1 was detected using the acetylcholinesterase (AChE) enzyme that is inhibited by this toxin, and the degree of inhibition was quantified by the Ellman's spectrophotometric method, obtaining a detection limit of 10 µg L(-1). OTA quantification was performed by monitoring its intrinsic fluorescence in methanol, reaching a detection limit of 0.1 µg L(-1). In order to successfully apply the analytical tool in the food analysis, immunoaffinity columns were used. Clean-up and quantification of both AFB1 and OTA in millet samples was obtained by HPLC-dedicated AflaOchra-Test HPLC™ (Vicam™) and Afla-OtaCLEAN™ (LC-Tech) immunoaffinity columns, followed by absorption/fluorescence detection. Millet samples which were fortified with both OTA (50 µg kg(-1)) and AFB1 (20 µg kg(-1)), gave recovery values of 100 ± 6% for OTA, and 110 ± 10% for AFB1, using AflaOchra-Test HPLC™. Single OTA clean-up and quantification in wine samples was obtained, using an OchraTest immunoaffinity column (Vicam™), reaching a detection limit of 0.3 µg L(-1) and recovery values between 80% and 120%. These results demonstrated the possibility of employing a single clean-up and a cost-effective, and easy to use analytical system for both AFB1 and OTA detection at µg kg(-1) (ppb) level. Furthermore, in the case of positive samples, they could be analysed further, using standard chromatographic procedures, without any additional clean-up step, since the same extraction procedure of standard method is proposed in our method. PMID:26838428

  3. Low cost quantitative digital imaging as an alternative to qualitative in vivo bioassays for analysis of active aflatoxin B1.

    Science.gov (United States)

    Rasooly, Reuven; Do, Paula M; Hernlem, Bradley J

    2016-06-15

    Aflatoxin B1 (AFB1) producing fungi contaminate food and feed and are a major health concern. To minimize the sources and incidence of AFB1 illness there is a need to develop affordable, sensitive mobile devices for detection of active AFB1. In the present study we used a low cost fluorescence detector and describe two quantitative assays for detection of detoxified and active AFB1 demonstrating that AFB1 concentration can be measured as intensity of fluorescence. When the assay plate containing increasing concentrations of AFB1 is illuminated with a 366 nm ultraviolet lamp, AFB1 molecules absorb photons and emit blue light with peak wavelength of 432 nm. The fluorescence intensity increased in dose dependent manner. However, this method cannot distinguish between active AFB1 which poses a threat to health, and the detoxified AFB1 which exhibits no toxicity. To measure the toxin activity, we used a cell based assay that makes quantification more robust and is capable of detecting multiple samples simultaneously. It is an alternative to the qualitative duckling bioassay which is the "gold-standard" assay currently being used for quantitative analysis of active AFB1. AFB1 was incubated with transduced Vero cells expressing the green fluorescence protein (GFP) gene. After excitation with blue light at 475 nm, cells emitted green light with emission peak at 509 nm. The result shows that AFB1 inhibits protein expression in a concentration dependent manner resulting in proportionately less GFP fluorescence in cells exposed to AFB1. The result also indicates strong positive linear relationship with R(2)=0.90 between the low cost CCD camera and a fluorometer, which costs 100 times more than a CCD camera. This new analytical method for measuring active AFB1 is low in cost and combined with in vitro assay, is quantitative. It also does not require the use of animals and may be useful especially for laboratories in regions with limited resources.

  4. Protective effects of baicalein and wogonin against benzo[a]pyrene- and aflatoxin B(1)-induced genotoxicities.

    Science.gov (United States)

    Ueng, Y F; Shyu, C C; Liu, T Y; Oda, Y; Lin, Y L; Liao, J F; Chen, C F

    2001-12-15

    To evaluate the protective effects of baicalein and wogonin against benzo[a]pyrene- and aflatoxin (AF) B(1)-induced toxicities, the effects of these flavonoids on the genotoxicities and oxidation of benzo[a]pyrene and AFB(1) were studied in C57BL/6J mice. Baicalein and wogonin reduced benzo[a]pyrene and AFB(1) genotoxicities as monitored by the umuC gene expression response in Salmonella typhimurium TA1535/pSK1002. Baicalein added in vitro decreased liver microsomal benzo[a]pyrene hydroxylation (AHH) activity with an ic(50) of 33.9 +/- 1.4 microM at 100 microM benzo[a]pyrene. Baicalein also inhibited AFQ(1) and AFB(1)-epoxide formation from AFB(1) (50 microM) oxidation (AFO) with ic(50) values of 22.8 +/- 1.4 and 5.3 +/- 0.8 microM, respectively. However, the in vitro inhibitory effects of wogonin on AHH and AFO activities in liver microsomes were less than those of baicalein as inhibition by 500 microM wogonin was only about 51-65%. Treatment of mice with liquid diets containing 5 mM baicalein and wogonin resulted in 22 and 49% decreases in hepatic AHH activities, respectively. Baicalein treatment resulted in 39 and 32% decreases in AFQ(1) and AFB(1)-epoxide formation from liver microsomal AFO, respectively. Wogonin treatment resulted in 39 and 47% decreases in AFQ(1) and AFB(1)-epoxide formation, respectively. A 1-week pretreatment with wogonin significantly decreased hepatic DNA adduct formation in mice treated with 200 mg/kg of benzo[a]pyrene via gastrogavage. These in vitro and in vivo effects suggested that baicalein and wogonin might have beneficial effects against benzo[a]pyrene- and AFB(1)-induced hepatic toxicities and that wogonin had a stronger protective effect in vivo. PMID:11755119

  5. Antioxidant efficacy of curcuminoids from turmeric ( Curcuma longa L.) powder in broiler chickens fed diets containing aflatoxin B1.

    Science.gov (United States)

    Gowda, Nisarani K S; Ledoux, David R; Rottinghaus, Goerge E; Bermudez, Alex J; Chen, Yin C

    2009-12-01

    A 3-week-feeding study (1-21 d post-hatch) was conducted to evaluate the efficacy of total curcuminoids (TCMN), as an antioxidant, to ameliorate the adverse effects of aflatoxin B1 (AFB1) in broiler chickens. Turmeric powder (Curcuma longa L.) that contained 2.55 % TCMN was used as a source of TCMN. Six cage replicates of five chicks each were assigned to each of six dietary treatments, which included: basal diet; basal diet supplemented with 444 mg/kg TCMN; basal diet supplemented with 1.0 mg/kg AFB1; basal diet supplemented with 74 mg/kg TCMN and 1.0 mg/kg AFB1; basal diet supplemented with 222 mg/kg TCMN and 1.0 mg/kg AFB1; basal diet supplemented with 444 mg/kg TCMN and 1.0 mg/kg AFB1. The addition of 74 and 222 mg/kg TCMN to the AFB1 diet significantly (P < 0.05) improved weight gain and feed efficiency. Increase (P < 0.05) in relative liver weight in birds fed AFB1 was significantly reduced (P < 0.05) with the addition of 74, 222 and 444 mg/kg TCMN to the AFB1 diet. The inclusion of 222 mg/kg TCMN ameliorated the adverse effects of AFB1 on serum chemistry in terms of total protein, albumin and gamma-glutamyl transferase activity. The decreased antioxidant functions due to AFB1 were also alleviated by the inclusion of 222 mg/kg TCMN. It is concluded that the addition of 222 mg/kg TCMN to the 1.0 mg/kg AFB1 diet demonstrated maximum antioxidant activity against AFB1.

  6. Intervention of Grape Seed Proanthocyanidin Extract on the Subchronic Immune Injury in Mice Induced by Aflatoxin B1

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    Miao Long

    2016-04-01

    Full Text Available The aim was to investigate the prevention of grape seed proanthocyanidin extract (GSPE on the subchronic immune injury induced by aflatoxin B1 (AFB1 and the possible ameliorating effect of GSPE in mice. The subchronic AFB1-induced immune injury mice model was set up with the continuous administration of 100 μg/kg body weight (BW AFB1 for six weeks by intragastric administration. Then, intervention with different doses (50 and 100 mg/kg BW of GSPE was conducted on mice to analyze the changes of body weight, immune organ index, antioxidant capability of spleen, serum immunoglobulin content, and the expression levels of inflammatory cytokines. The prevention of GSPE on the immune injury induced by AFB1 was studied. The GSPE could relieve the AFB1-induced reduction of body weight gain and the atrophy of the immune organ. The malondialdehyde (MDA level of the spleen in the AFB1 model group significantly increased, but levels of catalase (CAT, glutathione (GSH, glutathione peroxidase (GSH-PX, and superoxide dismutase (SOD significantly decreased. The GSPE could significantly inhibit the oxidative stress injury of the spleen induced by AFB1. AFB1 exposure could not significantly change the contents of IgA, IgG, or IgM. AFB1 significantly improved the expression of interleukin 1β (IL-1β, IL-6, tumor necrosis factor α (TNF-α, and interferon γ (IFN-γ. Additionally, GSPE could decrease the expression of these four proinflammatory factors to different degrees and inhibit the inflammatory reaction of mice. The results suggest that GSPE alleviates AFB1-induced oxidative stress and significantly improves the immune injury of mice induced by AFB1.

  7. Interactions between hepatitis B virus and aflatoxin B(1): effects on p53 induction in HepaRG cells.

    Science.gov (United States)

    Lereau, Myriam; Gouas, Doriane; Villar, Stéphanie; Besaratinia, Ahmad; Hautefeuille, Agnès; Berthillon, Pascale; Martel-Planche, Ghislaine; Nogueira da Costa, André; Ortiz-Cuaran, Sandra; Hantz, Olivier; Pfeifer, Gerd P; Hainaut, Pierre; Chemin, Isabelle

    2012-03-01

    Infection by hepatitis B virus (HBV) and dietary exposure to aflatoxin B(1) (AFB(1)) are the main risk factors for the development of chronic liver disease and hepatocellular carcinoma (HCC). How these factors cooperate is still largely unknown. AFB(1) activation leads to DNA adduction and mutagenesis, with a specific mutation at codon 249 in TP53 (p.R249S). So far, only limited studies have addressed the effects of AFB(1) on HBV replication. We have analysed the effects of both risk factors on p53 induction during HBV infection in HepaRG, a cell line with hepatocyte-like morphology that metabolizes AFB(1) and supports HBV infection. Exposure to AFB(1) up to 5 µM induced a downregulation of HBV replication after 48 h, as measured by a decrease in viral antigens in the culture medium (HBsAg, HBeAg and large envelope protein) and in intracellular levels of HBV transcripts, DNA and HBsAg. Conversely, HBV infection did not significantly modify AFB(1)-DNA adduct formation or repair as assessed by immunodot-blot assay, and the induction of p53 in response to AFB(1) was similar in infected and non-infected HepaRG cells. Overall, our results suggest that AFB(1) exposure decreases HBV replication, whereas DNA damage by AFB(1) and subsequent p53 induction is not affected by the presence of the virus. Thus, in HepaRG cell line, AFB(1) and HBV do not cooperate to increase DNA damage by AFB(1). Further studies on the effects of both factors in a context of chronicity are needed to better understand synergistic effects. PMID:22113009

  8. Natural occurrence of aflatoxin B1 in marketed foods and risk estimates of dietary exposure in Koreans.

    Science.gov (United States)

    Ok, Hyun Ee; Kim, Hyun Jung; Shim, Won Bo; Lee, Hyomin; Bae, Dong-Ho; Chung, Duck-Hwa; Chun, Hyang Sook

    2007-12-01

    Aflatoxin B1 (AFB1) is an unavoidable food contaminant. To evaluate the potential health risk of AFB1 to Koreans posed by food consumption, we determined the natural occurrence of AFB1 in food and estimated the excess risk for liver cancer through dietary exposure to AFB1. A total of 694 food samples collected from six different regions of South Korea were analyzed for their AFB, content. One hundred four of the 694 samples were found to give positive enzyme-linked immunosorbent assay (ELISA) readings for AFB1 and were further investigated with high-performance liquid chromatography. Thirty-two samples, including 2 maize samples, 3 soybean products, 20 peanut samples, nut samples, and their products, and 7 spices, were found to be contaminated with AFB1 (4.6% incidence), up to 48.6 microg kg(-1). The level of AFB1 contamination in 28 of the 32 food products was below 10 microg kg(-1), which is the legal tolerance limit in Korea. From data on daily food consumption, the exposure dose of AFB1 was estimated to be 6.42 x 10(-7) mg kg(-1) body weight (bw) day(-1). The major contributors to the dietary intake of AFB1 were soybean paste and soy sauce, which composed 91% of the total exposure to AFB1. The excess risk of liver cancer for those exposed to AFB1 through food intake was estimated to be 5.78 x 10(-6) for hepatitis B-negative individuals and 1.48 x 10(-4) for hepatitis B-positive individuals. These results suggest that special consideration is required to reduce the intake of AFB1 in hepatitis B-positive individuals.

  9. Identification of differentially expressed genes in aflatoxin B1-treated cultured primary rat hepatocytes and Fischer 344 rats.

    Science.gov (United States)

    Harris, A J; Shaddock, J G; Manjanatha, M G; Lisenbey, J A; Casciano, D A

    1998-08-01

    Aflatoxin B1 (AFB1), a mutagen and hepatocarcinogen in rats and humans, is a contaminant of the human food supply, particularly in parts of Africa and Asia. AFB1-induced changes in gene expression may play a part in the development of the toxic, immunosuppressive and carcinogenic properties of this fungal metabolite. An understanding of the-role of AFB1 in modulating gene regulation should provide insight regarding mechanisms of AFB1-induced carcinogenesis. We used three PCR-based subtractive techniques to identify AFB1-responsive genes in cultured primary rat hepatocyte RNA: differential display PCR (DD-PCR), representational difference analysis (RDA) and suppression subtractive hybridization (SSH). Each of the three techniques identified AFB1-responsive genes, although no individual cDNA was isolated by more than one technique. Nine cDNAs isolated using DD-PCR, RDA or SSH were found to represent eight genes that are differentially expressed as a result of AFB1 exposure. Genes whose mRNA levels were increased in cultured primary rat hepatocytes after AFB1 treatment were corticosteroid binding globulin (CBG), cytochrome P450 4F1 (CYP4F1), alpha-2 microglobulin, C4b-binding protein (C4BP), serum amyloid A-2 and glutathione S-transferase Yb2 (GST). Transferrin and a small CYP3A-like cDNA had reduced mRNA levels after AFB1 exposure. Full-length CYP3A mRNA levels were increased. When liver RNA from AFB1-treated male F344 rats was evaluated for transferrin, CBG, GST, CYP3A and CYP4F1 expression, a decrease in transferrin mRNA and an increase in CBG, GST, CYP3A and CYP4F1 mRNA levels was also seen. Analysis of the potential function of these genes in maintaining cellular homeostasis suggests that their differential expression could contribute to the toxicity associated with AFB1 exposure.

  10. Intervention of Grape Seed Proanthocyanidin Extract on the Subchronic Immune Injury in Mice Induced by Aflatoxin B1

    Science.gov (United States)

    Long, Miao; Zhang, Yi; Li, Peng; Yang, Shu-Hua; Zhang, Wen-Kui; Han, Jian-Xin; Wang, Yuan; He, Jian-Bin

    2016-01-01

    The aim was to investigate the prevention of grape seed proanthocyanidin extract (GSPE) on the subchronic immune injury induced by aflatoxin B1 (AFB1) and the possible ameliorating effect of GSPE in mice. The subchronic AFB1-induced immune injury mice model was set up with the continuous administration of 100 μg/kg body weight (BW) AFB1 for six weeks by intragastric administration. Then, intervention with different doses (50 and 100 mg/kg BW) of GSPE was conducted on mice to analyze the changes of body weight, immune organ index, antioxidant capability of spleen, serum immunoglobulin content, and the expression levels of inflammatory cytokines. The prevention of GSPE on the immune injury induced by AFB1 was studied. The GSPE could relieve the AFB1-induced reduction of body weight gain and the atrophy of the immune organ. The malondialdehyde (MDA) level of the spleen in the AFB1 model group significantly increased, but levels of catalase (CAT), glutathione (GSH), glutathione peroxidase (GSH-PX), and superoxide dismutase (SOD) significantly decreased. The GSPE could significantly inhibit the oxidative stress injury of the spleen induced by AFB1. AFB1 exposure could not significantly change the contents of IgA, IgG, or IgM. AFB1 significantly improved the expression of interleukin 1β (IL-1β), IL-6, tumor necrosis factor α (TNF-α), and interferon γ (IFN-γ). Additionally, GSPE could decrease the expression of these four proinflammatory factors to different degrees and inhibit the inflammatory reaction of mice. The results suggest that GSPE alleviates AFB1-induced oxidative stress and significantly improves the immune injury of mice induced by AFB1. PMID:27070584

  11. Intoxicação de bovinos por aflatoxina B1 presente em polpa cítrica: relato de um surto Aflatoxin B1 poisoning in bovines: an outbreak report

    Directory of Open Access Journals (Sweden)

    M.M. Melo

    1999-12-01

    Full Text Available O presente trabalho relata um surto de intoxicação pela aflatoxina B1 em 150 bovinos machos mestiços, provenientes de um rebanho de 1774 animais, em sistema de confinamento, ingerindo polpa cítrica peletizada comercial, 5kg por animal. Todos os animais foram avaliados clinicamente, e os 18 animais que morreram foram necropsiados e submetidos a exames histopatológicos. A intoxicação foi confirmada pela presença da aflatoxina B1 (5ppm em amostras da polpa cítrica peletizada utilizada na propriedade e em fragmentos de fígado dos animais que morreram.The present work reports an outbreak of intoxication by aflatoxin B1 in 150 adult bovine male, males from a herd of 1774 animals, in confinement system, fed on commercial citrus pellet. All animals were evaluated clinicaly, and ones that died were necropsied and tissues submitted to histopathotology. The intoxication was confirmed by aflatoxin B1 presence in samples of the citrus pellet and in fragments of liver of the animals which died.

  12. Determinação de aflatoxina B1 em pimenta (Piper nigrum L. e orégano (Origanum vulgare L. por cromatografia em camada delgada e densitometria Determination of aflatoxin B1 (Piper nigrum L. and oregano (Origanum vulgare L. by thin-layer chromatography and densitometry

    Directory of Open Access Journals (Sweden)

    Guilherme Prado

    2008-01-01

    Full Text Available An analytical study based on extraction with methanol-water, immunoaffinity cleanup and separation, identification and quantification of aflatoxin B1 by thin-layer chromatography,in ground black and white pepper and oregano was carried out. Validation of the applied methodology was done through accuracy and precision studies. Recoveries of aflatoxin B1 and relative standard deviations, from spice samples spiked at levels from 4.86 to 97.70 µg/kg, were, respectively, higher than 72% and lower than 20%. Application to spice samples available in Minas Gerais state, purchased at popular markets, showed no contamination with aflatoxin B1.

  13. The effect of humidity after gamma-irradiation on aflatoxin B-1 production of A.flavus in ground nutmeg and peanut

    Energy Technology Data Exchange (ETDEWEB)

    Hilmy, N.; Chosdu, R. [Center for the Application of Isotopes and Radiation, Jakarta (Indonesia); Matsuyama, A. [Tokyo University of Agriculture (Japan). Nodai Research Inst.

    1995-10-01

    The effect of humidity of 75 up to 97% after irradiation on radiosensitivity and aflatoxin B1 production of Aspergillus flavus isolated from Indonesian nutmeg were examined. Irradiation doses used were 0;0.5;1 and 3 kGy. Mould free ground nutmeg and peanut were used as the growth media, and about 10{sup 8} of spores were used to contaminate each of the media. Aflatoxin productions were measured after having incubated 3 days up to 5 months under humidity of 91 and 97%. Prior to HPLC analysis, aflatoxin was cleaned-up using an immunoaffinity column. The results were: (1) A. flavus indicated no or almost no growth under RH of 85% or less. (2) Under 91-97% RH, growth of mycelium and toxin production were inhibited more or less by irradiation up to 1 kGy, although the effectiveness of irradiation varied with different RH and media during postirradiation incubation. (3) By 3 kGy or more, both mycelium growth and toxin production of the mould were found to be completely inhibited. (4) The production of aflatoxin in nutmeg began after having incubated for 25 and 45 days and in peanut for 3 and 6 days under 97 and 91% RH, respectively. (Author).

  14. An ultra-high-performance liquid chromatography-tandem mass spectrometry method for simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2 in traditional Chinese medicines.

    Science.gov (United States)

    Han, Zheng; Zheng, Yunliang; Luan, Lianjun; Cai, Zengxuan; Ren, Yiping; Wu, Yongjiang

    2010-04-01

    An ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2 in traditional Chinese medicines (TCMs) was developed. The approach was characterized in details and a special focus was placed on the recovery rates of isolation procedure in different TCM matrices, i.e. rhizomes and roots, seeds, flowers, grasses and leaves. For this purpose, [(13)C(17)]-aflatoxinB1 was employed as the internal standard and a reliable solid phase extraction-based clean-up method was developed. The observed recovery rates of the six aflatoxins ranged from 85.6% to 117.6% in different matrices. Then, the established method was successfully applied to the determination of the six aflatoxins in various TCMs. For 30 commercial samples analyzed, 16 were contaminated with aflatoxins. The mean levels (incidence) of aflatoxins B1, B2, G1 and G2 in positive samples were 1.40 (68.8%), 1.27 (50.0%), 0.50 (43.8%) and 0.94 (43.8%) microg kg(-1), respectively. Interestingly, aflatoxin M1 was detected in two samples with the maximal content of 0.70 microg kg(-1). No sample was contaminated with aflatoxin M2. Meanwhile, a possible association between the contamination levels and the selected herbs was clarified in the present study. PMID:20363399

  15. 4种黄曲霉毒素B1酶联免疫试剂盒与液相法检测结果比较%A comparative study on 4 kinds of aflatoxin B1 assay kits and liquid chromatography

    Institute of Scientific and Technical Information of China (English)

    胡玲玲; 项瑜芝; 蔡增轩; 任一平

    2014-01-01

    Objective To compare and evaluate 4 kinds of aflatoxin B1 assay kits and liquid chromatography. Methods Three sample matrixes (infant formula rice flour, peanut butter and soy bean sauce) were detected using RIDASCREEN® aflatoxin B1 assay, AgraQuant® aflatoxin B1 assay, Helica low matrix aflatoxin B1 assay and Huaan magnech aflatoxin B1 assay, and then compared the analysis results with those by liquid chromatography. Results By contrast, Huaan magnech aflatoxin B1 assay applied to only the infant formula rice flour. HELICA low matrix aflatoxin B1 assay and RIDASCREEN® aflatoxin B1 assay could detect more sample matrixes, but the analysis results of Helica low matrix aflatoxin B1 assay were inconsistent with those by liquid chromatography, the background of RIDASCREEN® aflatoxin B1 assay was higher. AgraQuant® aflatoxin B1 assay were applied to peanut butter and soybean sauce. Conclusion In view of the different of the sample matrix, the suitable assay kit needed to be chosen to get the best analysis results. When the results of ELISA were positive, the HPLC method was needed for further validation.%目的:综合评价4种不同厂家的黄曲霉毒素 B1试剂盒。方法选择几种食品基质(婴幼儿米粉、花生酱、酱油),分别用R-Biopharm, Romer, Helica和华安麦科的黄曲霉毒素B1试剂盒检测,将测定结果与液相色谱法测得结果进行比对。结果通过与液相色谱法测得结果比较,华安麦科试剂盒只适合检测婴幼儿米粉样品。Helica和R-Biopharm试剂盒能检测多种基质,而Helica试剂盒检测结果与液相相符度低, R-Biopharm试剂盒检测米粉时,本底较高。Romer试剂盒适用于花生酱和酱油样品。结论为了得到最佳的检测结果,针对不同的样品基质,应选择合适的试剂盒。当用酶联法检测出阳性样品时,需用液相色谱法确认。

  16. Optimization of the Extraction Method of Aflatoxin B1 in Moon Cake%月饼中黄曲霉毒素B1检测前处理方法的改进

    Institute of Scientific and Technical Information of China (English)

    李姣; 佘之蕴; 林耀文; 刘海卿

    2012-01-01

    A study was conducted to optimize extraction method of Aflatoxin B1 from eight kinds of moon cake (mixed nuts, lotus seed with egg yolk, jujube paste, fruit, ham, snowy, and tuna) through ELISA. The detection results by two methods showed no significant difference except the detection in Mixed Nuts and Tuna. Better extraction efficacy was found using 50% methanol/water solution. The average recovery rate was 84.0%~l 14.7%. The method could improve the detection efficiency of aflatoxin B1 of moon cake.%通过对伍仁、蛋黄莲蓉、枣泥、水果、火腿、冰皮和吞拿鱼七种口味,共70批次月饼进行甲醇-水(1∶1,V/V)和三氯甲烷二次抽提两种处理方法对比,结果显示两种方法检测仅伍仁和吞拿鱼月饼差异显著,其余不显著.甲醇水提取方法检测结果重复性良好,加标回收率为84.0%114.7%,提高了月饼黄曲霉毒素B1的检测效率.

  17. Hematological Parameters and the State of Liver Cells of Rats After Oral Administration of Aflatoxin B1 Alone and Together with Nanodiamonds

    Science.gov (United States)

    Mogilnaya, O. A.; Puzyr, A. P.; Baron, A. V.; Bondar, V. S.

    2010-05-01

    Hematological parameters and the state of liver cells of rats were examined in vivo after the animals received aflatoxin B1 (AfB1) alone and together with modified nanodiamonds (MND) synthesized by detonation. The rats that had received the MND hydrosol had elevated leukocyte levels, mainly due to higher granulocyte counts and somewhat increased monocyte counts compared to control rats. Hematological parameters of the rats that had received AfB1 alone differed from those of the control rats in another way: total white blood cell counts were significantly lower due to the decreased lymphocyte counts. In rats that had consumed AfB1 with the MND hydrosol, changes in hematological parameters were less pronounced than in rats that had consumed either AfB1 or MND. Electron microscopy showed that hepatocytes of the rats that had received the MND hydrosol or AfB1 with the MND hydrosol contained elevated levels of lipid inclusions and lysosomes. Hyperplasia of the smooth endoplasmic reticulum (EPR) was revealed in liver specimens of the rats that had received AfB1. Results of the study suggest the conclusion about mutual mitigation of the effects of nanoparticles and the mycotoxin on rats blood and liver cells after AfB1 has adsorbed on MND.

  18. Hematological Parameters and the State of Liver Cells of Rats After Oral Administration of Aflatoxin B1 Alone and Together with Nanodiamonds

    Directory of Open Access Journals (Sweden)

    Baron AV

    2010-01-01

    Full Text Available Abstract Hematological parameters and the state of liver cells of rats were examined in vivo after the animals received aflatoxin B1 (AfB1 alone and together with modified nanodiamonds (MND synthesized by detonation. The rats that had received the MND hydrosol had elevated leukocyte levels, mainly due to higher granulocyte counts and somewhat increased monocyte counts compared to control rats. Hematological parameters of the rats that had received AfB1 alone differed from those of the control rats in another way: total white blood cell counts were significantly lower due to the decreased lymphocyte counts. In rats that had consumed AfB1 with the MND hydrosol, changes in hematological parameters were less pronounced than in rats that had consumed either AfB1 or MND. Electron microscopy showed that hepatocytes of the rats that had received the MND hydrosol or AfB1 with the MND hydrosol contained elevated levels of lipid inclusions and lysosomes. Hyperplasia of the smooth endoplasmic reticulum (EPR was revealed in liver specimens of the rats that had received AfB1. Results of the study suggest the conclusion about mutual mitigation of the effects of nanoparticles and the mycotoxin on rats blood and liver cells after AfB1 has adsorbed on MND.

  19. Glutathione-S-transferase A3 knockout mice are sensitive to acute cytotoxic and genotoxic effects of aflatoxin B1.

    Science.gov (United States)

    Ilic, Zoran; Crawford, Dana; Vakharia, Dilip; Egner, Patricia A; Sell, Stewart

    2010-02-01

    Aflatoxin B1 (AFB1) is a major risk factor for hepatocellular carcinoma (HCC) in humans. However, mice, a major animal model for the study of AFB1 carcinogenesis, are resistant, due to high constitutive expression, in the mouse liver, of glutathione S-transferase A3 subunit (mGSTA3) that is lacking in humans. Our objective was to establish that a mouse model for AFB1 toxicity could be used to study mechanisms of toxicity that are relevant for human disease, i.e., an mGSTA3 knockout (KO) mouse that responds to toxicants such as AFB1 in a manner similar to humans. Exons 3-6 of the mGSTA3 were replaced with a neomycin cassette by homologous recombination. Southern blotting, RT-PCR, Western blotting, and measurement of AFB1-N(7)-DNA adduct formation were used to evaluate the mGSTA3 KO mice. The KO mice have deletion of exons 3-6 of the mGSTA3 gene, as expected, as well as a lack of mGSTA3 expression at the mRNA and protein levels. Three hours after injection of 5 mg/kg AFB1, mGSTA3 KO mice have more than 100-fold more AFB1-N(7)-DNA adducts in their livers than do similarly treated wild-type (WT) mice. In addition, the mGSTA3 KO mice die of massive hepatic necrosis, at AFB1 doses that have minimal toxic effects in WT mice. We conclude that mGSTA3 KO mice are sensitive to the acute cytotoxic and genotoxic effects of AFB1, confirming the crucial role of GSTA3 subunit in protection of normal mice against AFB1 toxicity. We propose the mGSTA3 KO mouse as a useful model with which to study the interplay of risk factors leading to HCC development in humans, as well as for testing of additional possible functions of mGSTA3. PMID:19850059

  20. Isolation, screening and identification of Bacillus spp. as direct-fed microbial candidates for aflatoxin B1 biodegradation

    Institute of Scientific and Technical Information of China (English)

    Rosario Galarza-Seeber; Juan David Latorre; Xochitl Hernandez-Velasco; Amanda Drake Wolfenden; Lisa Renee Bielke; Anita Menconi; Billy Marshall Hargis; Guillermo Tellez

    2015-01-01

    To evaluate the ability of Bacillus spp. as direct-fed microbials (DFM) to biodegrade aflatoxin B1 (AFB1) by using an in vitro digestive model simulating in vivo conditions. Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 µg/mL of AFB1 in modified Czapek-Dox medium. Plates were incubated at 37 °C and observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16S rRNA sequence analyzes for identification. Tolerance to acidic pH, osmotic concentrations of NaCl, bile salts were tested, and antimicrobial sensitivity profiles were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time (3:15 h), supernatants and digesta were collected for high-performance liquid chromatography fluorescence detection analysis by triplicate. Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of fluorescence and area of clearance around each colony in modified Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identified the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions (pH 2.0), tolerant to a high osmotic pressure (NaCl at 6.5%), and were able to tolerate 0.037%bile salts after 24 h of incubation. No significant differences (P > 0.05) were observed in the concentrations of AFB1 in

  1. Phytic Acid Exposure Alters AflatoxinB1-induced Reproductive and Oxidative Toxicity in Albino Rats (Rattus norvegicus).

    Science.gov (United States)

    Abu El-Saad, Abdelaziz S; Mahmoud, Hamada M

    2009-09-01

    The increased use of feed in Egypt's aquaculture and animal industries raises concerns about the possible presence of mycotoxins in feedstuffs. The use of alternative medicine, such as botanicals and nutritional supplements, has become popular with inflammatory cases. The present study aimed to testify the role played by phytic acid (IP6) in enhancing the reproductive and oxidative toxicity induced in aflatoxinB1 (AFB1) treated white male albino rats (Rattus norvegicus) throughout treatment and withdrawal periods. One hundred and twenty white male albino rats were grouped into four groups. Group 1, was injected with 300 mug kg(-1) body wt of AFB1 once every 3 days for 15 days and left uninjected for another 15 days to study the withdrawal effect. Group 2, was injected with 300 mug kg(-1) body wt of AFB1 once every 3 days for 15 days and treated simultaneously with IP6 daily for another 15 days. Group 3, was treated daily with IP6 (40 mg kg(-1) body wt) for 15 days and with no treatment for other 15 days. Group 4, injected with equivalent volume of sterile phosphate buffer saline solution as a control group. Sera were taken at the experimental intervals and assayed for testosterone hormone, follicular-stimulating hormone (FSH) and luteinizing hormone (LH) to determine the toxicological impact of AFB1 and the possibility of amelioration by phytic acid on the reproductive performance of the studied animal. The effects of AFB1 treatment on the absolute and relative weight of testis as well as its histopathologic effect on the testis and the possibility of amelioration by IP6 treatment were evaluated. The activities of enzymatic and non-enzymatic anti-oxidants, in addition to lipid peroxidation were measured in the testis' homogenate of AFB1-treated rats. A decrease in sex hormone levels, an increase in testicular lipid peroxidation product levels and a significant decrease in testicular glutathione content, catalase and total peroxidase and superoxide dismutase

  2. Phytic Acid Exposure Alters AflatoxinB1-Induced Reproductive and Oxidative Toxicity in Albino Rats (Rattus norvegicus

    Directory of Open Access Journals (Sweden)

    Abdelaziz S. Abu El-Saad

    2009-01-01

    Full Text Available The increased use of feed in Egypt's aquaculture and animal industries raises concerns about the possible presence of mycotoxins in feedstuffs. The use of alternative medicine, such as botanicals and nutritional supplements, has become popular with inflammatory cases. The present study aimed to testify the role played by phytic acid (IP6 in enhancing the reproductive and oxidative toxicity induced in aflatoxinB1 (AFB1 treated white male albino rats (Rattus norvegicus throughout treatment and withdrawal periods. One hundred and twenty white male albino rats were grouped into four groups. Group 1, was injected with 300 μg kg−1 body wt of AFB1 once every 3 days for 15 days and left uninjected for another 15 days to study the withdrawal effect. Group 2, was injected with 300 μg kg−1 body wt of AFB1 once every 3 days for 15 days and treated simultaneously with IP6 daily for another 15 days. Group 3, was treated daily with IP6 (40 mg kg−1 body wt for 15 days and with no treatment for other 15 days. Group 4, injected with equivalent volume of sterile phosphate buffer saline solution as a control group. Sera were taken at the experimental intervals and assayed for testosterone hormone, follicular-stimulating hormone (FSH and luteinizing hormone (LH to determine the toxicological impact of AFB1 and the possibility of amelioration by phytic acid on the reproductive performance of the studied animal. The effects of AFB1 treatment on the absolute and relative weight of testis as well as its histopathologic effect on the testis and the possibility of amelioration by IP6 treatment were evaluated. The activities of enzymatic and non-enzymatic anti-oxidants, in addition to lipid peroxidation were measured in the testis’ homogenate of AFB1-treated rats. A decrease in sex hormone levels, an increase in testicular lipid peroxidation product levels and a significant decrease in testicular glutathione content, catalase and total peroxidase and superoxide

  3. Interactive effects of dietary protein concentration and aflatoxin B1 on performance, nutrient digestibility, and gut health in broiler chicks.

    Science.gov (United States)

    Chen, X; Naehrer, K; Applegate, T J

    2016-06-01

    A 20-day trial was conducted to determine the impact of aflatoxin B1 (AFB1) and dietary protein concentration on performance, nutrient digestibility, and gut health in broiler chicks. The 6 dietary treatments were arranged in a 2 × 3 factorial with 3 crude protein (CP) concentrations (16, 22, and 26%) with or without 1.5 mg/kg AFB1 Each diet was fed to 6 replicate cages (6 chicks per cage) from zero to 20 d of age. Endogenous N and amino acid loss were estimated from birds fed a N-free diet with or without 1.5 mg/kg AFB1 A significant interaction between AFB1 and CP concentration was observed for growth performance, where reduction of BW gain, feed intake, gain:feed ratio, and breast muscle weight by AFB1 were most profound in birds fed the 16%-CP diet, and were completely eliminated when birds were fed the 26%-CP diet (AFB1 by CP interaction; P ≤ 0.023). Similarly, AFB1 reduced serum albumin, total protein, and globulin concentrations in birds fed 16 and 22% CP diets, but not in those fed the 26%-CP (AFB1 by CP interaction; P ≤ 0.071). Gut permeability was increased in birds fed AFB1-contamiated diets as measured by serum lactulose/rhamnose ratio (main effect; P = 0.04). Additionally, AFB1 tended to increase endogenous N loss (P = 0.09), and significantly reduced apparent ileal digestible energy and standardized ileal N and amino acid digestibility in birds fed the 16%-CP diet, while birds fed higher dietary CP were not affected (AFB1 by CP interaction; P ≤ 0.01). Further, AFB1 increased the translation initiation factor 4E-binding protein (4EBP1), claudin1, and multiple jejunal amino acid transporters expression (main effect; P ≤ 0.04). Results from this study indicate that a 1.5 mg AFB1/kg diet significantly impairs growth, major serum biochemistry measures, gut barrier, endogenous loss, and energy and amino acid digestibility. Aflatoxicosis can be augmented by low dietary CP, while higher dietary CP completely eliminated the impairment of

  4. Glutathione-S-transferase A3 knockout mice are sensitive to acute cytotoxic and genotoxic effects of aflatoxin B1

    International Nuclear Information System (INIS)

    Aflatoxin B1 (AFB1) is a major risk factor for hepatocellular carcinoma (HCC) in humans. However, mice, a major animal model for the study of AFB1 carcinogenesis, are resistant, due to high constitutive expression, in the mouse liver, of glutathione S-transferase A3 subunit (mGSTA3) that is lacking in humans. Our objective was to establish that a mouse model for AFB1 toxicity could be used to study mechanisms of toxicity that are relevant for human disease, i.e., an mGSTA3 knockout (KO) mouse that responds to toxicants such as AFB1 in a manner similar to humans. Exons 3-6 of the mGSTA3 were replaced with a neomycin cassette by homologous recombination. Southern blotting, RT-PCR, Western blotting, and measurement of AFB1-N7-DNA adduct formation were used to evaluate the mGSTA3 KO mice. The KO mice have deletion of exons 3-6 of the mGSTA3 gene, as expected, as well as a lack of mGSTA3 expression at the mRNA and protein levels. Three hours after injection of 5 mg/kg AFB1, mGSTA3 KO mice have more than 100-fold more AFB1-N7-DNA adducts in their livers than do similarly treated wild-type (WT) mice. In addition, the mGSTA3 KO mice die of massive hepatic necrosis, at AFB1 doses that have minimal toxic effects in WT mice. We conclude that mGSTA3 KO mice are sensitive to the acute cytotoxic and genotoxic effects of AFB1, confirming the crucial role of GSTA3 subunit in protection of normal mice against AFB1 toxicity. We propose the mGSTA3 KO mouse as a useful model with which to study the interplay of risk factors leading to HCC development in humans, as well as for testing of additional possible functions of mGSTA3.

  5. Determinação de aflatoxina B1 em pimenta (Piper nigrum L.) e orégano (Origanum vulgare L.) por cromatografia em camada delgada e densitometria Determination of aflatoxin B1 (Piper nigrum L.) and oregano (Origanum vulgare L.) by thin-layer chromatography and densitometry

    OpenAIRE

    Guilherme Prado; Marize Silva de Oliveira; Ana Paula Aprígio Moreira; Adriana de Souza Lima; Robson de Assis Souza; Matheus do Carmo Alves

    2008-01-01

    An analytical study based on extraction with methanol-water, immunoaffinity cleanup and separation, identification and quantification of aflatoxin B1 by thin-layer chromatography,in ground black and white pepper and oregano was carried out. Validation of the applied methodology was done through accuracy and precision studies. Recoveries of aflatoxin B1 and relative standard deviations, from spice samples spiked at levels from 4.86 to 97.70 µg/kg, were, respectively, higher than 72% and lower ...

  6. Study on the Technique for Rapid Detection of Aflatoxin B 1 in Stored Grain%储粮中黄曲霉毒素B1快速检测技术的研究

    Institute of Scientific and Technical Information of China (English)

    鲍会梅

    2014-01-01

    Content by high performance liquid chromatography method for the detection of aflatoxin B 1 in stored grain,the test sample of the content of aflatoxin B1 0.004 3 mg/kg,the sample RSD was 0.033,the recovery rate was 97.41%,extract was acetonitrile∶water=84∶16,mobile phase of methanol∶acetonitrile∶water was mixed solu-tion=15∶13∶72,linear regression equation was y=188.97x+4.96,correlation coefficient was R2=0.999 8,the excita-tion wavelength of=460 nm detector,using derivative method photochemical derivatization pool,evolutionary method using immunoaffinity column. HPLC method for the determination of aflatoxin B 1 was a method for detect-ing had the advantages of simple operation,good accuracy,high sensitivity.%运用高效液相色谱方法检测储粮中黄曲霉毒素B1的含量,检测样品中黄曲霉毒素B1的含量为0.0043 mg/kg,样品的RSD为0.033,回收率为97.41%,提取液是乙腈∶水=84∶16,流动相是甲醇∶乙腈∶水混合溶液=15∶13∶72,线性回归方程是y=188.97x+4.96相关系数是R2=0.9998,检测器激发波长=460 nm,衍生法利用光化学衍生池,进化方法利用免疫亲和柱。高效液相色谱法测定黄曲霉毒素B1是一种操作简单、准确度好、灵敏度高的检测方法。

  7. 柱前衍生-高效液相色谱法测定茶叶中的黄曲霉毒素 B1%HPLC Determination of Aflatoxin B1 in Tea with Pre-column Derivatization

    Institute of Scientific and Technical Information of China (English)

    郭爱华; 王玮; 李小丽

    2014-01-01

    HPLC was applied to the determination of aflatoxin B1 in tea with pre-column derivatization.The sample was extracted with acetonitrile (85 + 15 )solution.The supernatant was purified with MycoSepTM 226 column,then derivatized by adding n-hexane and trifluoroacetic acid.C18 column was used as stationary phase and the analyte was determined with fluorescence detector.Linear relationship between values of peak area and mass concentration of aflatoxin B1 was kept in the range of 0.20-10.0 μg·L-1 with detection limit (3S/N)of 0.1 μg· kg-1 .Tests for recovery were made at the concentration levels of 0.5,1.0 and 5.0 μg ·L-1 of aflatoxin B1 standard,giving values of recovery and RSD′s (n = 6)in the ranges of 91.9% - 102% and 1.5% - 6.9%respectively.%应用柱前衍生-高效液相色谱法测定茶叶中黄曲霉毒素 B1的含量。样品采用乙腈(85+15)溶液提取,滤液用 MycoSepTM226柱净化,加入正己烷和三氟乙酸衍生,经 C18色谱柱分离,荧光检测器检测。黄曲霉毒素 B1的质量浓度在0.20~10.0μg·L-1范围内与其峰面积呈线性关系,检出限(3S/N)为0.1μg·kg-1。在0.5,1.0,5.0μg·L-1等3个浓度水平进行加标回收试验,回收率在91.9%~102%之间,测定值的相对标准偏差(n=6)在1.5%~6.9%之间。

  8. 高效液相色谱法测定潮州花生糖中的黄曲霉毒素B1%Determination of Aflatoxin B1 in Chaozhou Peanut Brittle by High Performance Liquid Chromatography

    Institute of Scientific and Technical Information of China (English)

    谢鸿; 杨泽川; 陈旭明

    2014-01-01

    A method was developed for the determination of Aflatoxin B1 in Chaozhou peanut brittle by Immunoaffinity chromatography and High Performance Liquid Chromatography. Samples extracted with methanol- water (7+3), filtered with qualitative filter paper and glass fiber filter paper, purified by immunoaffinity column;Eluate was dried with nitrogen, and derived by n-hexane and trifluoroacetic acid, dissolved with water-acetonitrile (85+15) and detected by high performance liquid chromatography with fluorescence detection. The calibration curves are linear between 0.002 5 ng/μL and 0.100 ng/μL. The rate of average recovery of aflatoxin B 1 was 85.3%-92.9%,and RSDs ranged from 0.61%to 1.08%. In this paper,optimized the condition of HPLC and the conditions of pre-column derivatization, the method has the advantages of simplicity,rapidity,precision,good reproducibility and high recovery,so that it could be used for the determination of aflatoxin B1, and it can meet the need of measuring in food safety control.%建立免疫亲和层析净化-高效液相色谱法测定潮州花生糖中的黄曲霉毒素B1的检测方法。样品用甲醇-水(7+3)提取,定性滤纸和玻璃纤维滤纸2次过滤,过免疫亲和柱净化;洗脱液氮气吹干后加正己烷和三氟乙酸衍生,以水-乙腈(85+15)溶解,用高效液相色谱仪带荧光检测器检测。方法在0.0025 ng/μL~0.100 ng/μL的范围内呈良好的线性关系,加标平均回收率为85.3%~92.9%,试验的RSD为0.61%~1.08%。该方法通过改进柱前衍生化的条件和优化仪器分析条件,实验结果重现性好,回收率高,结果准确,为黄曲霉毒素B1的检测提供了准确可靠的方法。

  9. Effect of aflatoxin B1 on growth and enzymatic activity of a native strain of Bacillus sp Efecto de la aflatoxina B1 sobre el crecimiento y actividad proteolítica de una cepa nativa de Bacillus sp

    Directory of Open Access Journals (Sweden)

    Márquez Edna Judith

    2004-07-01

    Full Text Available The effect of different aflatoxin B1 (AFAB1 concentrations on alkaline protease growth and enzymatic activity was evaluated; a native strain of alkalophilic Bacillus sp cultivated in CSL (Corn Steep Liquor was used. It was found that the effect of AFAB1 on the strain inhibited its growth and enzymatic activity to 1 ppm, showing that the strain is highly sensible to AFAB1, meaning that medium obtained f rom Colombian corn contaminated with this mycotoxin cannot be easily used. Concentrations less than 0.1 ppm did not affect growth and enzymatic activity. Key words: Bacillus, aflatoxin, alkaline proteases.Se evaluó el efecto de diferentes concentraciones de aflatoxina B1 (AFAB1 sobre el crecimiento y actividad enzimática de proteasas alcalinas de una cepa nativa de Bacillus sp Alcalofílico cultivada en LAM (Licor Agotado de Maíz. Se encontró que la cepa inhibe su crecimiento y actividad enzimática a 1 ppm, lo que demuestra una alta sensibilidad de la cepa evaluada a la AFAB1 e imposibilita utilizar fácilmente medios obtenidos de maíz nacional contaminado con esta micotoxina. Las concentraciones inferiores a 0.1 ppm no tienen ningún efecto sobre el crecimiento y la actividad enzimática. Palabras clave: Bacillus, aflatoxina, proteasas alcalinas.

  10. A preliminary study on the occurrence of Aspergillus spp. and aflatoxin B1 in imported wheat and barley in Penang, Malaysia.

    Science.gov (United States)

    Reddy, K R N; Salleh, Baharuddin

    2010-11-01

    Thirty samples consisting of wheat (15) and barley (15) were collected from different markets in Penang, Malaysia, originating from India and Thailand, respectively. All samples were analyzed for occurrence of Aspergillus spp. and aflatoxin B1 (AFB1). Aspergillus flavus was dominant in all samples followed by A. niger. AFB1 could be detected in three wheat samples ranging from 0.42 to 1.89 μg/kg and one barley sample had 0.58 μg/kg of AFB1. The AFB1 levels in all the samples were below the Malaysian regulatory limits (Penang, Malaysia.

  11. Ameliorative Effects of Tinospora Cordifolia Root Extract on Histopathological and Biochemical Changes Induced by Aflatoxin-B1 in Mice Kidney

    OpenAIRE

    Gupta, Rekha; Sharma, Veena

    2011-01-01

    The present study was planned to investigate the ability of the Tinospora cordifolia to scavenge free radicals generated during aflatoxicosis. A total no. of 48 male Swiss albino mice (30 ± 5 g) were exposed to Aflatoxin B1(AFB1) (2 μg/30 g b.wt, orally) either individually or in combination with T. cordifolia (50, 100, 200 mg/kg, orally) once daily for 25 days. AFB1 exposure led to significant rise in thiobarbituric acid reactive substances (TBARS) and fall in superoxide dismutase (SOD), cat...

  12. Performance Improvement of the One-Dot Lateral Flow Immunoassay for Aflatoxin B1 by Using a Smartphone-Based Reading System

    OpenAIRE

    Jihea Moon; Giyoung Kim; Sangdae Lee

    2013-01-01

    This study was conducted to develop a simple, rapid, and accurate lateral flow immunoassay (LFIA) detection method for point-of-care diagnosis. The one-dot LFIA for aflatoxin B1 (AFB1) was based on the modified competitive binding format using competition between AFB1 and colloidal gold-AFB1-BSA conjugate for antibody binding sites in the test zone. A Smartphone-based reading system consisting of a Samsung Galaxy S2 Smartphone, a LFIA reader, and a Smartphone application for the image acquisi...

  13. Effects of Milk Yield, Feed Composition, and Feed Contamination with Aflatoxin B1 on the Aflatoxin M1 Concentration in Dairy Cows’ Milk Investigated Using Monte Carlo Simulation Modelling

    Directory of Open Access Journals (Sweden)

    H. J. van der Fels-Klerx

    2016-10-01

    Full Text Available This study investigated the presence of aflatoxin M1 (AfM1 in dairy cows’ milk, given predefined scenarios for milk production, compound feed (CF contamination with aflatoxin B1 (AfB1, and inclusion rates of ingredients, using Monte Carlo simulation modelling. The model simulated a typical dairy farm in the Netherlands. Six different scenarios were considered, based on two lactation and three CF composition scenarios. AfB1 contamination of the CF was based on results from the Dutch national monitoring programme for AfB1 in feed materials from 2000 until 2010. Monitoring data from feed materials used in CF production for dairy cattle in the Netherlands were used. Additionally, AfB1 contamination data from an incident in maize in 2013 were used. In each scenario, five different transfer equations of AfB1 from feed to AfM1 in the milk were used, and 1000 iterations were run for each scenario. The results showed that under these six scenarios, the weekly farm concentration of AfM1 in milk was above the EC threshold in less than 1% of the iterations, with all five transfer equations considered. However, this increased substantially in weeks when concentrations from the contaminated maize batch were included, and up to 28.5% of the iterations exceeded the EC threshold. It was also observed that an increase in the milk production had a minimal effect on the exceedance of the AfM1 threshold due to an apparent dilution effect. Feeding regimes, including the composition of CF and feeding roughages of dairy cows, should be carefully considered based on the potential AfM1 contamination of the farm’s milk.

  14. An attempt to model the probability of growth and aflatoxin B1 production of Aspergillus flavus under non-isothermal conditions in pistachio nuts.

    Science.gov (United States)

    Aldars-García, Laila; Ramos, Antonio J; Sanchis, Vicente; Marín, Sonia

    2015-10-01

    Human exposure to aflatoxins in foods is of great concern. The aim of this work was to use predictive mycology as a strategy to mitigate the aflatoxin burden in pistachio nuts postharvest. The probability of growth and aflatoxin B1 (AFB1) production of aflatoxigenic Aspergillus flavus, isolated from pistachio nuts, under static and non-isothermal conditions was studied. Four theoretical temperature scenarios, including temperature levels observed in pistachio nuts during shipping and storage, were used. Two types of inoculum were included: a cocktail of 25 A. flavus isolates and a single isolate inoculum. Initial water activity was adjusted to 0.87. Logistic models, with temperature and time as explanatory variables, were fitted to the probability of growth and AFB1 production under a constant temperature. Subsequently, they were used to predict probabilities under non-isothermal scenarios, with levels of concordance from 90 to 100% in most of the cases. Furthermore, the presence of AFB1 in pistachio nuts could be correctly predicted in 70-81 % of the cases from a growth model developed in pistachio nuts, and in 67-81% of the cases from an AFB1 model developed in pistachio agar. The information obtained in the present work could be used by producers and processors to predict the time for AFB1 production by A. flavus on pistachio nuts during transport and storage.

  15. Structural Analysis and Biological Toxicity of Aflatoxins B1 and B2 Degradation Products Following Detoxification by Ocimum basilicum and Cassia fistula Aqueous Extracts

    Science.gov (United States)

    Iram, Wajiha; Anjum, Tehmina; Iqbal, Mazhar; Ghaffar, Abdul; Abbas, Mateen; Khan, Abdul Muqeet

    2016-01-01

    This study showed the comparison between Ocimum basilicum and Cassia fistula (leaves and branch) aqueous extracts for their ability to detoxify of aflatoxins B1 and B2 (AFB1; 100 μg L-1 and AFB2; 50 μg L-1) by In Vitro assays and decontamination studies. Results indicated that O. basilicum leaves extract was found to be highly significant (P aflatoxins, under optimized conditions, i.e., pH 8, temperature 30°C and incubation period of 72 h. Moreover the antifungal activity of these plants extracts were also tested. The findings depicted that O. basilicum leaves extract showed maximum growth inhibition of aflatoxigenic isolates, i.e., 82–87% as compared to other tested plants extracts. The structural elucidation of degraded toxin products by LCMS/MS analysis showed that nine degraded products of AFB1 and AFB2 were formed. MS/MS spectra showed that most of the products were formed by the removal of double bond in the terminal furan ring and modification of lactone group indicating less toxicity as compared to parent compounds. Brine shrimps bioassay further confirmed the low toxicity of degraded products, showing that O. basilicum leaves extract can be used as an effective tool for the detoxification of aflatoxins. PMID:27471501

  16. Structural Elucidation and Toxicity Assessment of Degraded Products of Aflatoxin B1 and B2 by Aqueous Extracts of Trachyspermum ammi.

    Science.gov (United States)

    Iram, Wajiha; Anjum, Tehmina; Iqbal, Mazhar; Ghaffar, Abdul; Abbas, Mateen

    2016-01-01

    In this study aqueous extract of seeds and leaves of Trachyspermum ammi were evaluated for their ability to detoxify aflatoxin B1 and B2 (AFB1; 100 μg L(-1) and AFB2; 50 μg L(-1)) by in vitro and in vivo assays. Results indicated that T. ammi seeds extract was found to be significant (P < 0.05) in degrading AFB1 and AFB2 i.e., 92.8 and 91.9% respectively. However, T. ammi leaves extract proved to be less efficient in degrading these aflatoxins, under optimized conditions i.e., pH 8, temperature 30°C and incubation period of 72 h. The structural elucidation of degraded toxin products by LCMS/MS analysis showed that eight degraded products of AFB1 and AFB2 were formed. MS/MS spectra showed that most of the products were formed by the removal of double bond in the terminal furan ring and modification of lactone group indicating less toxicity as compared to parent compounds. Brine shrimps bioassay further confirmed the low toxicity of degraded products, showing that T. ammi seeds extract can be used as an effective tool for the detoxification of aflatoxins.

  17. Structural elucidation and toxicity assessment of degraded products of aflatoxin B1 and B2 by aqueous extracts of Trachyspermum ammi

    Directory of Open Access Journals (Sweden)

    Wajiha eIram

    2016-03-01

    Full Text Available In this study aqueous extract of seeds and leaves of Trachyspermum ammi were evaluated for their ability to detoxify aflatoxin B1 and B2 (AFB1; 100 µg L-1 and AFB2; 50 µg L-1 by In Vitro and In Vivo assays. Results indicated that T. ammi seeds extract was found to be highly significant (P < 0.05 in degrading AFB1 and AFB2 i.e. 92.8% and 91.9% respectively. However T. ammi leaves extract proved to be less efficient in degrading these aflatoxins, under optimized conditions i.e., pH 8, temperature 30˚C and incubation period of 72h. The structural elucidation of degraded toxin products by LCMS/MS analysis showed that eight degraded products of AFB1 and AFB2 were formed. MS/MS spectra showed that most of the products were formed by the removal of double bond in the terminal furan ring and modification of lactone group indicating less toxicity as compared to parent compounds. Brine shrimps bioassay further confirmed the low toxicity of degraded products, showing that T. ammi seeds extract can be used as an effective tool for the detoxification of aflatoxins.

  18. Survey For Detection and Determination of Aflatoxins M1 and B1 in local Milk and Certain Dairy Products by Thin Layer Chromatographic Method

    Directory of Open Access Journals (Sweden)

    T.A. Nassib *,S.N. Guergues** , M.M. Motawee

    2005-03-01

    Full Text Available 90 different type of milk samples, 10 Yogurt Samples, 110 different type of cheese samples and 10 ice cream samples were collected randomly from Giza Governorate during the summer of 1998 ­ 1999, for detection and determination of Aflatoxins M1& B1 by using thin layer chromatographic method. Results revealed that the average range of Aflatoxin M1 in milk samples amounted from 0.144 to 0.378 ng/ml. About 20 % of cows and buffaloes milk samples contained form 0.378 to 0.342 ng/ml of AFM1, whereas about 10% of other milk samples were contaminated with 0.162, 0.288, 0.324, 0.234, 0.144 and 0.162 ng/ml for skim , Pasteurized , sterilized, UHT, powder, and baby milk, in the same order. Concentrations of AFM1 detected in cheese samples, furthermore, varied due to the type and age of cheese being examined. 20% of cheese samples were contaminated with AFM1 being 5.1, 3.2, 2.99, 2.099, and 2.34 ng/gm for fresh Domiati, aged Domiati, Processed and Karish cheese, respectively, whereas, 30% of the other types of cheese contained 5.88, 6.3 and 3.4 ng/gm for Roquefort, fresh Romi, and Cheddar cheese, respectively. The lowest concentration of AFM1, of 0.116 ng/gm was detected, however, in 10% of yogurt samples. Meanwhile, 20% of ice cream samples were found to be contaminated with 2.7 ng/ml, and 10% of Feta cheese samples contained 3.3 ng/gm. It could also be appeared from results that both of cream and spread cheese were found completely free from this Aflatoxin, the lowest content of Aflatoxin detected in all of the above examined samples was 0.116, 0.162, 0.162 and 0.216 (ppb in yogurt, skim, baby milk and cream, respectively. On the other hand, results also indicated that all milk samples were free from Aflatoxin B1 except one sample of skim milk (out of 10 which gave positive result.

  19. Application of dispersive liquid-liquid microextraction for the determination of aflatoxins B1, B2, G1 and G2 in cereal products.

    Science.gov (United States)

    Campone, Luca; Piccinelli, Anna Lisa; Celano, Rita; Rastrelli, Luca

    2011-10-21

    The application of dispersive liquid-liquid microextraction (DLLME) technique for the rapid analysis of aflatoxins B(1), B(2), G(1) and G(2) in maize, rice and wheat products has been evaluated. After extraction of aflatoxins from cereal matrices with a mixture of methanol/water 8:2 (v/v), the analytes were rapidly transferred from the extract to another small volume of organic solvent, chloroform, by DLLME. Aflatoxins were determined using high performance liquid chromatography with florescence detection and photochemical post-column derivatization. Parameters affecting both extraction and DLLME procedures, such as extraction solvent, type and volume of DLLME extractant, volume of water and salt effect, were systematically investigated and optimized to achieve the best extraction efficiency. Under the optimal experimental conditions, the whole analytical method provides enrichment factors around 2.5 times and detection limits (0.01-0.17 μg kg(-1)) below the maximum levels imposed by current regulation for aflatoxins in cereals and cereal products intended for direct human consumption. Recoveries (67-92%) and repeatability (RSD<10, n=3), tested in three different cereal matrices, meet the performance criteria required by EC Regulation No. 401/2006 for the determination of the levels of mycotoxins in foodstuffs. The proposed method was successfully applied to the analysis of retail cereal products with quantitative results comparable to the immunoaffinity chromatography (IAC). The main advantages of developed method are the simplicity of operation, the rapidity to achieve a very high sample throughput and low cost. PMID:21636088

  20. Bioactivation of aflatoxin B1 by lipoxygenases, prostaglandin H synthase and cytochrome P450 monooxygenase in guinea-pig tissues.

    Science.gov (United States)

    Liu, L; Massey, T E

    1992-04-01

    In the present investigation, we have examined the role of lipoxygenases in the bioactivation of aflatoxin B1 (AFB1) in hepatic and extrahepatic tissues. The enzyme activities were evaluated by determining [3H]AFB1-DNA adduct formation. The results demonstrated that both purified soybean lipoxygenase and guinea-pig tissue cytosolic lipoxygenases were able to activate AFB1 to form [3H]AFB1-DNA adduct(s). The reaction was completely inhibited by nordihydroguaiaretic acid (NDGA, 0.1 mM), a lipoxygenase inhibitor and an antioxidant, but not by indomethacin (0.1 mM), an inhibitor of prostaglandin H synthase (PHS), indicating that this reaction is associated with lipoxygenase activity, and/or is involved in a peroxyl radical process. While purified lipoxygenase showed arachidonic acid (AA)-dependent properties, the omission of AA did not diminish guinea-pig tissue cytosolic [3H]AFB1-DNA adduct formation, possibly because AA was released from lipid particles by AFB1. Within the range of hemoglobin (Hb) concentrations found in lung, kidney and liver cytosols (1.4-11.1 microM) and microsomes (0-0.5 microM), neither pure Hb, nor Hb of cytosols or microsomes from whole blood caused detectable AA-dependent AFB1-DNA binding. This indicates that Hb, as a contaminant with quasi-lipoxygenase activity, did not contribute to AFB1 activation attributed to guinea-pig tissue lipoxygenases. [3H]AFB1 concentrations at half-maximal DNA binding rate of pulmonary cytochrome P450 monooxygenases (P450) and lipoxygenases were similar, though P450 had a much higher maximum DNA binding rate. Pulmonary microsomal PHS activity for AFB1 activation was too low for its half-maximal binding concentrations of [3H]AFB1 and maximum rate to be accurately determined. In kidney, maximum rates for lipoxygenase, PHS and P450 were similar, whereas half-maximal binding concentrations for reactions by lipoxygenase and P450 were lower compared to that of PHS. The half-maximal binding concentration of hepatic

  1. Validation of a confirmatory analytical method for the determination of aflatoxins B1, B2, G1 and G2 in foods and feed materials by HPLC with on-line photochemical derivatization and fluorescence detection

    OpenAIRE

    Muscarella, Marilena; Iammarino, Marco; Nardiello, Donatella; Lo Magro, Sonia; Palermo, Carmen; Centonze, Diego; Palermo, Domenico

    2009-01-01

    Abstract A sensitive and selective analytical method for the simultaneous separation and quantitative determination of aflatoxins B1, B2, G1 and G2 in foodstuffs and materials for feed has been validated. The method is based on high performance liquid chromatography with on-line post-column photochemical derivatization and fluorescence detection. The chromatographic separation of aflatoxins was accomplished by using a C18 column eluted with an isocratic mobile phase consisting of w...

  2. Alterações hepáticas em codornas japonesas submetidas à intoxicação prolongada por aflatoxina B1 Hepatic changes in japanese quail after long term intoxication by aflatoxin B1

    Directory of Open Access Journals (Sweden)

    Carlos Augusto Fernandes Oliveira

    2004-02-01

    Full Text Available O objetivo do presente trabalho foi estudar os efeitos da aflatoxina B1 (AFB1 sobre as vísceras (fígado, baço e moela de codornas poedeiras japonesas, em condições de exposição a baixas doses, tendo em vista que são poucos os dados de toxicidade de longa duração nesta espécie. Assim, foram constituídos 4 grupos formados, cada um, por 6 codornas de linhagem comercial, as quais receberam rações contendo AFB1 nas concentrações de 0 (controle, 25, 50 e 100mg.kg-1, durante 168 dias. As aves do grupo 100mg kg_1 apresentaram fígados com peso relativo médio menor (p The aim of the present record was to study the effects of aflatoxin B1 (AFB1 on selected viscera (liver, spleen and gizzard of laying Japanese quail under conditions of low level exposure, in view of the little information regarding the long term toxicity on this specie. Thus, four experimental groups of six commercial quails were constituted and given rations containing either 0 (controls, 25, 50 or 100mg aflatoxin B1 (AFB1/kg feed, during 168 days. When compared to controls, birds from group 100mg.kg-1 presented low relative liver weight (p < 0.05. Histological changes were observed only in the livers, and all samples from quail exposed to AFB1 revealed moderate to severe hepatic cell vacuolation with fatty change, particularly in birds from groups receiving highest levels of toxin (50 and 100mg.kg-1. Bile duct hyperplasia occurred only in the birds exposed to 100mg.kg-1 of AFB1. The results indicated that long term administration of AFB1 at levels above 50mg.kg-1 can cause significant hepatic lesions in Japanese quail.

  3. 无需衍生化样品处理快速测定食品中黄曲霉毒素B1%Rapid determination of aflatoxin B1 in foods without derivation

    Institute of Scientific and Technical Information of China (English)

    李芳; 黎琳琳; 刘绪斌; 沙力; 李艳美; 储晓刚; 粟有志; 雷红琴

    2014-01-01

    目的:建立无需衍生化法-超高效液相色谱直接测定食品中的黄曲霉毒素 B1的快速检测方法。方法样品经甲醇-水(7:3, v:v)提取,免疫亲和柱净化, C18色谱柱分离后,采用大体积流通池荧光检测器检测。结果黄曲霉毒素B1在0.10~5.00 ng/mL范围内与峰面积呈良好线性关系, R2>0.999,在饼干、粉丝、点心、沙琪玛、蛋糕、辣椒酱6种基质中,黄曲霉毒素B1在0.50~2.00μg/kg范围内加标,平均回收率在71.7%~111.2%之间,相对标准偏差(RSDs)为3.8%~11.5%,检出限(LOD)为0.05μg/kg。结论本方法灵敏、快速、无衍生、结果准确,能够满足食品中黄曲霉毒素B1残留的检测需求。%Objective To establish a rapid determination of aflatoxin B1 in food by ultra performance liquid chromatography (UPLC) without derivation method. Methods Samples were extracted with methanol water (7:3, v:v), cleaned up by immuno-affinity column, separated by C18 column, and determined by fluorescence detector with large volume flow cell. Results High correlation coefficient (r2>0.999) was obtained in the linear range of 0.10~5.00 ng/mL. The average recovery rates were 71.7%~111.2% at the spiked level of 0.50~2.00 μg/kg and relative standard deviation(RSD) was 3.8%~11.5% in biscuit, vermicelli, refreshment, sacima, cakes, and chili sauce. The limit of quantitation (LOD) was 0.05μg/kg. Conclusion This method is sensitive, rapid, non-derived, and accurate, and it can meet the requirements for determination of aflatoxin B1 in foods.

  4. Extraction of aflatoxin B1 by ultrasonic radiation%应用超声波法提取大米中黄曲霉毒素B1

    Institute of Scientific and Technical Information of China (English)

    孙秀兰; 张银志

    2007-01-01

    目的:建立一种超声波快速提取大米中黄曲霉毒素B1的方法,并对提取方法的回收率和准确度进行评价.方法:采用ELISA方法,研究和评价了超声波提取大米中黄曲霉素B1的提取效率.结果:与普通振荡处理相比,采用超声波处理明显增大了提取效率,缩短了提取时间,超声波处理10 min时达到最大提取率,振荡处理30 min才能达到,而且超声波最大提取浓度比振荡处理提高了30.5%.在针对0.1、1和10μg/kg这3个浓度水平做加标回收试验,回收率达到83.0%~95.2%,比振荡处理提高10%左右.结论:超声频率20 KHz,时间为10 min,为提取黄曲霉毒素B1的最佳条件,而且不同超声频率对结果产生的影响不显著.

  5. Degradation of Aflatoxin B 1 by Microwave-lightwave Combination in Peanut%微波光波组合辐射去除花生表面黄曲霉毒素B1的研究

    Institute of Scientific and Technical Information of China (English)

    赵越; 王瑞琦; 刘睿杰; 李秋; 王珊珊; 杜润鸿; 常明

    2014-01-01

    针对花生表面存在的黄曲霉毒素污染问题,考察了微波光波组合辐射处理对花生表面黄曲霉毒素B1(AFB1)的降解效果。将花生在一定微波光波辐射频率下进行试验,研究结果表明:微波光波联合辐射法可有效降解花生表面的AFB1,并随着时间增加,毒素减少量也增加,8min时降解率达到74.06%; AFB1降解速率与样品最初被污染的水平有关;经该法处理后的花生,其基本成分变化不大,营养成分损失较少。由于微波光波组合辐射法降解条件温和、操作简单、降解效率高,可应用于受黄曲霉毒素污染的粮食物料中。%For problem of contaminated peanut with aflatoxin, effect of microwave-lightwave combined treatment on degradation of aflatoxin B1 in peanut was studied. Results of experiment indicated that method of microwave and light-wave combined treatment could effectively degrade AFB1 in peanut, degradation level of AFB1 in peanut increased with treatment time, and under condition of 8 min, degradation rate of AFB1 was 74.06%. Degradation rate of AFB1 was relat-ed to initial level of contamination. Basic ingredients of peanut changed little, and less nutrition was lost. Since method was under soft condition, simple and efficient, it could be applied to the crops contaminated with aflatoxin.

  6. Structural Analysis and Biological Toxicity of Aflatoxins B1 and B2 Degradation Products Following Detoxification by Ocimum basilicum and Cassia fistula Aqueous Extracts.

    Science.gov (United States)

    Iram, Wajiha; Anjum, Tehmina; Iqbal, Mazhar; Ghaffar, Abdul; Abbas, Mateen; Khan, Abdul Muqeet

    2016-01-01

    This study showed the comparison between Ocimum basilicum and Cassia fistula (leaves and branch) aqueous extracts for their ability to detoxify of aflatoxins B1 and B2 (AFB1; 100 μg L(-1) and AFB2; 50 μg L(-1)) by In Vitro assays and decontamination studies. Results indicated that O. basilicum leaves extract was found to be highly significant (P < 0.05) in degrading AFB1 and AFB2, i.e., 90.4 and 88.6%, respectively. However, O. basilicum branch, C. fistula leaves and branch extracts proved to be less efficient in degrading these aflatoxins, under optimized conditions, i.e., pH 8, temperature 30°C and incubation period of 72 h. Moreover the antifungal activity of these plants extracts were also tested. The findings depicted that O. basilicum leaves extract showed maximum growth inhibition of aflatoxigenic isolates, i.e., 82-87% as compared to other tested plants extracts. The structural elucidation of degraded toxin products by LCMS/MS analysis showed that nine degraded products of AFB1 and AFB2 were formed. MS/MS spectra showed that most of the products were formed by the removal of double bond in the terminal furan ring and modification of lactone group indicating less toxicity as compared to parent compounds. Brine shrimps bioassay further confirmed the low toxicity of degraded products, showing that O. basilicum leaves extract can be used as an effective tool for the detoxification of aflatoxins. PMID:27471501

  7. [Determination of aflatoxin B1, B2, G1, G2 in armeniacae semen amarum by high-performance liquid chromatography-tandem mass spectrometry].

    Science.gov (United States)

    Zheng, Run-Sheng; Xu, Hui; Wang, Wen-Li; Zhan, Ruo-Ting; Chen, Wei-Wen

    2013-10-01

    A simple, rapid and cost-effective high-performance liquid chromatography-tandem mass spectrometry (LC-MS/ MS) method was established for simultaneous determination of aflatoxins (AFB1, AFB2, AFG1, AFG2) in Armeniacae Semen Amarum and the application was performance in 11 samples collected from different markets, medical stores and hospitals. The sample was extracted with 84% acetonitrile/water and 250 microL extraction was directly injected into a LC-MS/MS system without further purification procedure after being redissolved with methanol. The LC separation was performed on a C18 column with a linear gradient elution program of 4 mmol x L(-1) NH4 Ac-0.1% formic acid solution and menthol as the mobile phase. Selected reaction monitoring (SRM) was used for selective determination of the four aflatoxins on a triple quadruple mass spectrometer, which was operated in positive ionization modes. All the four aflatoxins showed a good linear relationship with r > 0.999 0, the average recoveries were between 87.88% and 102.9% and the matrix effect was ranged from 90.71% to 99.30% in low, intermediate and high levels. Furthermore, the higher recovery was obtained by the method reported in this study, comparing to the cleanup procedure with the Mycosep 226 purification column. Eleven samples collected were detected and the contamination levels of the AFB1 were between 1.590-2.340 microg x kg(-1) and the AF (B1 + B2 + G1 + G2) was ranged from 2.340 to 3.340 microg x kg(-1). In summary, the developed method was suitable to detect and screen AFB1, AFB2, AFG1, AFG2 in Armeniacae Semen Amarum. PMID:24490568

  8. Aflatoxins B1, B2, G1, and G2 contamination in ground red peppers commercialized in Sanliurfa, Turkey.

    Science.gov (United States)

    Karaaslan, Mehmet; Arslanğray, Yusuf

    2015-04-01

    Aflatoxins (AFs) are hepatogenic, teratogenic, imunosuppressive, and carcinogenic fungal metabolites found in feeds, nuts, wine-grapes, spices, and other grain crops. Humans are exposed to AFs via consumption of mycotoxin-contaminated foods. This study aimed to determine the prevalence of AF contamination in powdered red peppers sold in Sanliurfa. A total of 42 samples were randomly collected from retail shops, supermarkets, open bazaars, and apiaries and examined for the occurrence and levels of AFB1, AFB2, AFG1, and AFG2 toxins. AFs were determined by using an HPLC system after pre-separation utilizing immunoaffinity columns. AFs levels were below 2.5 μg/kg in 16 samples, between 2.5 and 10 μg/kg in 13 samples while 13 samples had AFs higher than the tolerable limit (10 μg/kg) according to the regulations of Turkish Food Codex and European Commission. The occurrence of AF fractions during powdered red pepper processing steps was also evaluated. According to the results obtained in this study, it was found that the highest AF accumulations in powdered red peppers start during perspiration and final drying of the products processed on soil contacted surfaces while there was no limit exceeding aflatoxin contamination in the samples produced on concrete surfaces.

  9. Size-dependent modulation of graphene oxide-aptamer interactions for an amplified fluorescence-based detection of aflatoxin B1 with a tunable dynamic range.

    Science.gov (United States)

    Zhang, JingJing; Li, Zengmei; Zhao, Shancang; Lu, Yi

    2016-06-20

    Aflatoxin B1 (AFB1) is a common toxin found in many foods. While AFB1 sensors have been reported, few studies have shown amplified detection with tunable dynamic ranges. We herein report a simple and highly sensitive amplified aptamer-based fluorescent sensor for AFB1, which relies on the ability of nano-graphene oxide (GO) to protect aptamers from nuclease cleavage for amplified detection and on the nanometer size effect of GO to tune the dynamic range and sensitivity. The assay was performed by simply mixing the carboxyl-X-rhodamine (ROX)-labeled AFB1 aptamer, the GO, the nuclease, and the AFB1 samples. Modulating the size of the GO nanosheet resulted in three dynamic ranges, i.e., 12.5 to 312.5 ng mL(-1), 1.0 to 100 ng mL(-1), and 5.0 to 50 ng mL(-1), with corresponding limits of detection of 10.0 ng mL(-1), 0.35 ng mL(-1) and 15.0 ng mL(-1), respectively. The sensor was highly selective against other aflatoxins and common molecules in foods, and its performance was verified in corn samples spiked with known concentration of AFB1. PMID:27137348

  10. Aflatoxin B1 Detection Using a Highly-Sensitive Molecularly-Imprinted Electrochemical Sensor Based on an Electropolymerized Metal Organic Framework

    Directory of Open Access Journals (Sweden)

    Mengjuan Jiang

    2015-09-01

    Full Text Available A sensitive electrochemical molecularly-imprinted sensor was developed for the detection of aflatoxin B1 (AFB1, by electropolymerization of p-aminothiophenol-functionalized gold nanoparticles in the presence of AFB1 as a template molecule. The extraction of the template leads to the formation of cavities that are able to specifically recognize and bind AFB1 through π-π interactions between AFB1 molecules and aniline moities. The performance of the developed sensor for the detection of AFB1 was investigated by linear sweep voltammetry using a hexacyanoferrate/hexacyanoferrite solution as a redox probe, the electron transfer rate increasing when the concentration of AFB1 increases, due to a p-doping effect. The molecularly-imprinted sensor exhibits a broad linear range, between 3.2 fM and 3.2 µM, and a quantification limit of 3 fM. Compared to the non-imprinted sensor, the imprinting factor was found to be 10. Selectivity studies were also performed towards the binding of other aflatoxins and ochratoxin A, proving good selectivity.

  11. Pathological changes in the immune organs of broiler chickens fed on corn naturally contaminated with aflatoxins B1 and B2.

    Science.gov (United States)

    Peng, Xi; Bai, Shiping; Ding, Xuemei; Zeng, Qiufeng; Zhang, Keying; Fang, Jing

    2015-01-01

    The purpose of this study was to evaluate the underlying basis for aflatoxin-induced immunosuppression in the broiler chicken by detecting pathological lesions and apoptosis in thymus, bursa of Fabricius (BF) and spleen. COBB500™ male broiler chicks were randomly allocated to two groups. The control group was fed on a basal corn-based diet while the other group (the AFB group) was fed on a similar diet but the corn was naturally contaminated with aflatoxins B1 and B2. Histopathological examination revealed that in the AFB group there was more nuclear debris in the three immune organs and obvious congestion of red pulp in the spleen, when compared with the control group. Ultrastructural examination showed lesions in the lymphocytes and reticulocytes of the three immune organs, the mucosal epithelium of the BF and the plasmocytes of the spleen. Increased apoptotic cells and an impaired membrane system (including nuclear membrane, mitochondria and endoplasmic reticulum [ER]) could be observed in the three immune organs in birds of the AFB group. In the plasmocytes, dilated rough endoplasmic reticulum contained electron-dense matrix. By flow cytometry, the percentages of apoptosis were significantly higher (P < 0.01) in the three organs of the AFB group than those of the control group. These observations suggested that the lesions of the immune organs were related to the immunosuppression, and that the apoptosis might be initiated by the mitochondrial pathway and ER chaperone pathway.

  12. Determination of aflatoxins B1, B2, G1, G2 and ochratoxin A in animal feed by ultra high-performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    López Grío, Sergio José; Garrido Frenich, Antonia; Martínez Vidal, José Luis; Romero-González, Roberto

    2010-03-01

    A rapid and simple method was developed to determine aflatoxins B1, B2, G1, G2 and ochratoxin A in animal feed and pet foods by UHPLC-MS/MS. Because the complexity of the evaluated matrices, the proposed method is based on sonication extraction using an ACN/water mixture (80:20 v/v) followed by a clean-up step utilising C(18) as sorbent. Performance parameters of the method were evaluated, including linearity, trueness, precision and LOQ. Good linearity was found for all mycotoxins, with determination coefficients higher than 0.99 in the range considered, using matrix-matched calibration for quantification purposes. Recoveries ranged from 84 to 113%, with RSD lower than 20%, whereas LOQs were 5 microg/kg for the assayed mycotoxins. Finally, the method was successfully applied to the analysis of 19 real samples, detecting aflatoxin G2 in two samples at 13 and 17 microg/kg respectively, whereas the other mycotoxins were detected at trace levels (

  13. Aflatoxin B1 Detection Using a Highly-Sensitive Molecularly-Imprinted Electrochemical Sensor Based on an Electropolymerized Metal Organic Framework.

    Science.gov (United States)

    Jiang, Mengjuan; Braiek, Mohamed; Florea, Anca; Chrouda, Amani; Farre, Carole; Bonhomme, Anne; Bessueille, Francois; Vocanson, Francis; Zhang, Aidong; Jaffrezic-Renault, Nicole

    2015-09-01

    A sensitive electrochemical molecularly-imprinted sensor was developed for the detection of aflatoxin B1 (AFB1), by electropolymerization of p-aminothiophenol-functionalized gold nanoparticles in the presence of AFB1 as a template molecule. The extraction of the template leads to the formation of cavities that are able to specifically recognize and bind AFB1 through π-π interactions between AFB1 molecules and aniline moities. The performance of the developed sensor for the detection of AFB1 was investigated by linear sweep voltammetry using a hexacyanoferrate/hexacyanoferrite solution as a redox probe, the electron transfer rate increasing when the concentration of AFB1 increases, due to a p-doping effect. The molecularly-imprinted sensor exhibits a broad linear range, between 3.2 fM and 3.2 µM, and a quantification limit of 3 fM. Compared to the non-imprinted sensor, the imprinting factor was found to be 10. Selectivity studies were also performed towards the binding of other aflatoxins and ochratoxin A, proving good selectivity. PMID:26371042

  14. Effect of Milk Thistle (Silybum marianum L. on Biochemical Parameters and Immunity of Broiler Chicks Fed Aflatoxin B1 after Three Weeks

    Directory of Open Access Journals (Sweden)

    Maliheh Amiri Dumari

    2014-09-01

    Full Text Available Background: This study was conducted to determine the efficacy of milk thistle seeds (MTSs in counteracting the toxic effects of aflatoxin B1 (AFB1 in a contaminated diet fed to broilers. Methods: Two dietary inclusion rates of AFB1 (0, 0.250 and 500 ppb and MTS (0, 0.5 and 1% were tested in a 3×3 factorial manner. The effect of nine experimental treatments was assessed using 216 one-d-old Ross 308 male broiler chicks in a randomized complete design with 4 replicates of 6 birds each from one to 21 days of age. The effects of dietary AFB1 and MTS on serum biochemistry factors, antibody titer against Newcastle disease (ND and influenza disease (ID in broilers were evaluated at the end of this period. Results: Statistical analysis of the main effects of diets indicated no significant changes in uric acid, cholesterol, triglycerides, low density lipoprotein (LDL, ID, and phosphorus compared to the control (P>0.01. Also, addition of 500 ppb of dietary AFB1 into the diet was associated with significant decreases in serum glucose, calcium, high density lipoprotein (HDL, and ND compared to the control group (P<0.01. The contaminated diet significantly increased the activities of aspartate aminotransferase (AST and alanine aminotransferase (ALT (P<0.05. Conclusion: Milk thistle showed protective effects and resulted in some serum enzyme activities and serum biochemical changes associated with aflatoxin toxicity.

  15. Assessment of isorhamnetin 3-O-neohesperidoside from Acacia salicina: protective effects toward oxidation damage and genotoxicity induced by aflatoxin B1 and nifuroxazide.

    Science.gov (United States)

    Bouhlel, Ines; Limem, Ilef; Skandrani, Ines; Nefatti, Aicha; Ghedira, Kamel; Dijoux-Franca, Marie-Genevieve; Leila, Chekir-Ghedira

    2010-08-01

    Antioxidant activity of isorhamnetin 3-O-neohesperidoside, isolated from the leaves of Acacia salicina, was determined by the ability of this compound to inhibit xanthine oxidase activity and to scavenge the free radical 2,2'-azino-bis (3-ethylbenzthiazoline-6-sulfonic acid) (ABTS(.-)) diammonium salt. Antigenotoxic activity was assessed using the SOS chromotest assay. This compound has the ability to scavenge the ABTS(.+) radical by a hydrogen donating mechanism. We also envisaged the study of the antioxidant effect of this compound by the enzymatic xanthine/xanthine oxidase (X/XOD) assay. Results indicated that isorhamnetin 3-O-neohesperidoside was a potent inhibitor of xanthine oxidase and superoxide anion scavengers. Moreover, this compound induced an inhibitory activity against nifuroxazide and aflatoxine B1 (AFB1) induced genotoxicity. Taken together, these observations provide evidence that isorhamnetin 3-O-neohesperidoside isolated from the leaves of A. salicina is able to protect cells against the consequences of oxidative stress. PMID:20809543

  16. Effects of mycotoxins in cultured kidney cells: cytotoxicity of aflatoxin B1 in Madin-Darby and primary fetal bovine kidney cells.

    Science.gov (United States)

    Yoneyama, M; Sharma, R P; Elsner, Y Y

    1987-04-01

    The cytotoxicity of aflatoxin B1, a fungal metabolite and an important food contaminant, was evaluated in an established cell line, Madin-Darby bovine kidney (MDBK) cells, and in primary fetal bovine kidney (PFBK) cells. Cells were grown in monolayers and treated with media containing AFB1 for 24 hr. Both culture systems were sensitive to the chemical; the PFBK cultures were approximately four times more susceptible to the cytotoxic effect. Cell multiplication decreased in both systems in long-term cultures. Adherence of MDBK cells was only slightly reduced at the toxic concentrations of the chemical. Electron microscopy revealed condensation of chromatin, separation of nuclei from the cytoplasm, cytoplasmic vacuolization, and loss of surface microvilli. Results indicated differential sensitivity of the two culture systems.

  17. Comparative study of in vitro prooxidative properties and genotoxicity induced by aflatoxin B1 and its laccase-mediated detoxification products.

    Science.gov (United States)

    Zeinvand-Lorestani, Hamed; Sabzevari, Omid; Setayesh, Neda; Amini, Mohsen; Nili-Ahmadabadi, Amir; Faramarzi, Mohammad Ali

    2015-09-01

    In this paper, the enzymatic detoxification of aflatoxin B1 (AFB1) by laccase was studied, and the prooxidant properties and mutagenicity of the detoxification products were compared with those of AFB1. The optimal enzymatic reaction occurred in 0.1M of citrate buffer containing 20% DMSO at 35 °C, a pH of 4.5, and a laccase activity of 30 U mL(-1). After 2 d, sixty-seven percent of the toxic substrate was removed. The prooxidative properties of the detoxified products (27% versus 86%) and the mutagenicity were significantly decreased in comparison with the parent toxin. Unlike AFB1, which promoted metabolism-dependent genetic mutations by base-pair substitution, the detoxified products did not induce genotoxicity. Comparison of the Km values for AFB1 and riboflavin, a valuable food nutrient, indicated that laccase showed greater affinity for the toxin than for riboflavin. PMID:25876029

  18. Determination of aflatoxin B1 in cereals by homogeneous liquid-liquid extraction coupled to high performance liquid chromatography-fluorescence detection.

    Science.gov (United States)

    Sheijooni-Fumani, Neda; Hassan, Jalal; Yousefi, Seyed R

    2011-06-01

    A simple and rapid method based on homogeneous liquid-liquid extraction coupled to HPLC with fluorescence detection was developed for the determination of aflatoxin B1 (AFB1) in the rice and grain samples after post-column derivatization. The proposed method eliminated the use of immunoaffinity columns for clean-up in the determination of AFB1. The parameters affecting recovery and preconcentration such as type and volume of organic solvent, volume ratio of water/methanol, concentration of phase separator reagent and extraction time were optimized. Under the optimized conditions, the calibration graph was linear in the concentration range of 0.01-1.0 ng/g with the detection limit of 0.003 ng/g. This method was successfully applied for the analysis of AFB1 in different cereal samples. PMID:21491592

  19. Potential natural exposure of endangered red-crowned crane (Grus japonensis) to mycotoxins aflatoxin B1, deoxynivalenol, zearalenone, T-2 toxin, and ochratoxin A.

    Science.gov (United States)

    Liu, Da-wei; Liu, Hong-yi; Zhang, Hai-bin; Cao, Ming-chang; Sun, Yong; Wu, Wen-da; Lu, Chang-hu

    2016-02-01

    A survey was conducted to determine whether mycotoxins were present in the foods consumed by red-crowned cranes (Grus japonensis) in the Yancheng Biosphere Reserve, China. Collected in the reserve's core, buffer, and experimental zones during overwintering periods of 2013 to 2015, a total of 113 food samples were analyzed for aflatoxin B1, deoxynivalenol, zearalenone, T-2 toxin, and ochratoxin A using high performance liquid chromatography (HPLC). The contamination incidences vary among different zones and the mycotoxins levels of different food samples also presented disparity. Average mycotoxin concentration from rice grain was greater than that from other food types. Among mycotoxin-positive samples, 59.3% were simultaneously contaminated with more than one toxin. This study demonstrated for the first time that red-crowned cranes were exposed to mycotoxins in the Yancheng Biosphere Reserve and suggested that artificial wetlands could not be considered good habitats for the birds in this reserve, especially rice fields.

  20. Antioxidant activity and inhibition of aflatoxin B1-, nifuroxazide-, and sodium azide-induced mutagenicity by extracts from Rhamnus alaternus L.

    Science.gov (United States)

    Ammar, Rebai Ben; Sghaier, Mohamed Ben; Boubaker, Jihed; Bhouri, Wissem; Naffeti, Aicha; Skandrani, Ines; Bouhlel, Ines; Kilani, Soumaya; Ghedira, Kamel; Chekir-Ghedira, Leila

    2008-07-10

    The effect of extracts obtained from Rhamnus alaternus L. leaves on genotoxicity and SOS response induced by aflatoxin B(1) (10 microg/assay) as well as nifuroxazide (20 microg/assay) was investigated in a bacterial assay system, i.e., the SOS chromotest with Escherichia coli PQ37. The evaluation of the mutagenic and antimutagenic actions of the same extracts against the sodium azide (1.5 microg/plate)-induced mutagenicity was assayed using the Salmonella typhimurium assay system. The R. alaternus tested extracts exhibited no genotoxicity either with or without the external S9 activation mixture. However, all the extracts, particularly aqueous extract (A) and its chloroformic fraction (A(2)) significantly decreased the genotoxicity induced by aflatoxin B(1) and nifuroxazide. Moreover, the different extracts showed no mutagenicity when tested with Salmonella typhimurium strains TA1535 and TA1538 either with or without the S9 mix. Aqueous extract as well as its A(2) fraction exhibited the highest level of protection towards the direct mutagen, sodium azide-induced response in TA1535 strain with mutagenicity inhibition percentages of 83.6% and 91.4%, respectively, at a dose of 250 microg/plate. The results obtained by the Ames test assay confirm those of SOS chromotest. These same active extracts exhibited high xanthine oxidase (XOD) inhibiting with respective IC(50) values of 208 and 137 microg/ml, and superoxide anion-scavenging effects (IC(50) values of 132 and 117 microg/ml) when tested in the XOD enzymatic assay system. Our findings emphasize the potential of R. alaternus to prevent mutations and also its antioxidant effect. PMID:18511029

  1. Simultaneous determination of aflatoxin B(1), B(2), G(1), G(2), ochratoxin A, and sterigmatocystin in traditional Chinese medicines by LC-MS-MS.

    Science.gov (United States)

    Zheng, Runsheng; Xu, Hui; Wang, Wenli; Zhan, Ruoting; Chen, Weiwen

    2014-05-01

    In this paper we describe a rapid, simple, and costeffective liquid chromatography–tandem mass spectrometric (LC–MS–MS) method for simultaneous analysis of aflatoxin B1, B2, G1, and G2, ochratoxin A, and sterigmatocystin in 25 traditional Chinese medicines (TCMs). The method is based on single extraction with 84:16 (v/v) acetonitrile–water then analysis of the diluted crude extract without further clean-up. Chromatographic separation was achieved on a C18 column, with a mobile phase gradient prepared from aqueous 4 mmol L−1 ammonium acetate–0.1 % formic acid and methanol. Quantification of the analytes was by selective reaction monitoring (SRM) on a triple-quadrupole mass spectrometer in positive-ionization mode. Special focus was on investigating and reducing matrix effects to improve accuracy. The established method was validated by determination of linearity (r>0.995), sensitivity (limits of quantification 1.6–25.0 ng L−1), apparent recovery (84.8–110.6 %), extraction recovery (83.6–106.1 %), and precision (relative standard deviation ≤9.9 %) for two representative TCMs, Semen Armeniacae Amarae and Radix Pseudostellariae. The applicability of the method to TCMs other than these was further investigated, and 23 other TCMs with acceptable matrix effects (80.2–118.6 %) were screened. The validated method was finally used to assess mycotoxin contamination of 244 samples of 25 TCMs collected from local hospitals and TCM pharmacies. Aflatoxin B1 and ochratoxin A were detected in 5.3 % of the samples. Sterigmatocystin, the most prevalent mycotoxin contaminant, was present in 26.2 % of the samples tested; this has not been reported previously. The results of this work imply greater attention should be devoted to evaluation of the potential hazard caused by sterigmatocystin in TCMs.

  2. Simultaneous determination of aflatoxin B(1), B(2), G(1), G(2), ochratoxin A, and sterigmatocystin in traditional Chinese medicines by LC-MS-MS.

    Science.gov (United States)

    Zheng, Runsheng; Xu, Hui; Wang, Wenli; Zhan, Ruoting; Chen, Weiwen

    2014-05-01

    In this paper we describe a rapid, simple, and costeffective liquid chromatography–tandem mass spectrometric (LC–MS–MS) method for simultaneous analysis of aflatoxin B1, B2, G1, and G2, ochratoxin A, and sterigmatocystin in 25 traditional Chinese medicines (TCMs). The method is based on single extraction with 84:16 (v/v) acetonitrile–water then analysis of the diluted crude extract without further clean-up. Chromatographic separation was achieved on a C18 column, with a mobile phase gradient prepared from aqueous 4 mmol L−1 ammonium acetate–0.1 % formic acid and methanol. Quantification of the analytes was by selective reaction monitoring (SRM) on a triple-quadrupole mass spectrometer in positive-ionization mode. Special focus was on investigating and reducing matrix effects to improve accuracy. The established method was validated by determination of linearity (r>0.995), sensitivity (limits of quantification 1.6–25.0 ng L−1), apparent recovery (84.8–110.6 %), extraction recovery (83.6–106.1 %), and precision (relative standard deviation ≤9.9 %) for two representative TCMs, Semen Armeniacae Amarae and Radix Pseudostellariae. The applicability of the method to TCMs other than these was further investigated, and 23 other TCMs with acceptable matrix effects (80.2–118.6 %) were screened. The validated method was finally used to assess mycotoxin contamination of 244 samples of 25 TCMs collected from local hospitals and TCM pharmacies. Aflatoxin B1 and ochratoxin A were detected in 5.3 % of the samples. Sterigmatocystin, the most prevalent mycotoxin contaminant, was present in 26.2 % of the samples tested; this has not been reported previously. The results of this work imply greater attention should be devoted to evaluation of the potential hazard caused by sterigmatocystin in TCMs. PMID:24658469

  3. The bio-control of aflatoxin B1 contamination by fungi%真菌对黄曲霉毒素B1污染的防治研究

    Institute of Scientific and Technical Information of China (English)

    徐丹; 雷娇; 康敏; 宋宏新; 肖瑜; 石伟力

    2016-01-01

    黄曲霉毒素是由黄曲霉、寄生曲霉等曲霉属菌株所分泌的毒性次级代谢产物,其中,尤以黄曲霉毒素B1(Aflatoxin B1,AFB1)的毒性、致突变性、致癌性最强,而且其在农作物的栽培、收获、贮藏和加工过程中污染严重,受到全世界的广泛关注.因此,为保证食品的安全性,各国研究人员一直都在寻求安全、高效、经济、环保的方法来控制食品和饲料中AFB1的污染.近年来真菌在AFB1的生物防治方面已取得很大进展,并有部分菌株已应用于生产中.本研究就真菌对AFB1的防治机制及其前景展望进行综述.

  4. An in vitro study of alkaline phosphatase sensitivity to mixture of aflatoxin B1 and fumonisin B1 in the hepatopancreas of coastal lagoon wild and farmed shrimp Litopenaeus vannamei.

    Science.gov (United States)

    Pérez-Acosta, Jesús A; Burgos-Hernandez, Armando; Velázquez-Contreras, Carlos A; Márquez-Ríos, Enrique; Torres-Arreola, Wilfrido; Arvizu-Flores, Aldo A; Ezquerra-Brauer, J Marina

    2016-08-01

    This study aimed to establish the combined effect of aflatoxin B1 (AFB1) and fumonisin B1 (FB1) on wild Litopenaeus vannamei hepatopancreas alkaline phosphatase (AP) activity compared with that of farmed shrimp. AP activity in hepatopancreas extract was confirmed by several specific inhibitor assays. AP activity of wild shrimp was higher than that of farmed shrimp (p shrimp was inhibited when incubated with AFB1 and FB1. The greatest inhibition occurred when AP was incubated with a mixture of AFB1 and FB1. The IC50 for AFB1 on AP activity of wild and farmed shrimp hepatopancreases was 0.790 and 0.398 μg/mL, respectively. The IC50 of FB1 was 0.87 μg/mL for wild shrimp and 0.69 μg/mL for farmed shrimp. These results suggest that, at the mycotoxins concentrations used in the study, AP from farmed L. vannamei was sensitive to the presence of both mycotoxins; however, AP is more sensitive to the combination of AFB1 + FB1 suggesting a possible synergistic or potentiating inhibitory effect.

  5. 不同前处理方法对饲草中黄曲霉毒素B1测定的影响%Effect of Different Pretreatments on The Determination of AflatoxinB1 in Forage Grass

    Institute of Scientific and Technical Information of China (English)

    张海涛; 徐燕飞; 袁艺; 龚燕; 周韬; 周清莹; 林慧; 张东升

    2013-01-01

    本试验以样品加标回收率为指标,比较了液-液萃取、直接提取、亲和层析三种前处理方法对饲草中黄曲霉毒素B1测定值准确性的影响.应用酶联免疫法(ELISA)进行测定,并通过液相色谱荧光检测法(HPLC)进行验证.结果,采用液-液萃取-ELISA法,平均回收率在70.0%~102.0%,且相对标准偏差(RSD)<6.6%,亲和层析净化后ELISA与HPLC结果基本一致,但较液-液萃取测定值低.提示液-液萃取法作为ELISA检测饲草中黄曲霉毒素B1的前处理方法为最佳方法.%To determine the best pretreatment of aflatoxin B1 in forage by ELISA,this paper compared liquidliquid extraction,direct extraction and immunoaffinity chromatography with standard recovery rate of samples,and then verified by HPLC with fluorescence detection.As for liquid-liquid extraction,the average recovery rate is 70.0%~102.0%,and relative standard deviation(RSD) is less than 6.6%.ELISA is roughly identical to the rusults of HPLC,but it is lower than liquid-liquid extraction.The result showed that,it is feasible and accurate to determine AFB1 by liquid-liquid extraction pretreatment method in forage.

  6. Resistência de quatro genótipos de amendoim à produção de aflatoxina B1 após inoculação com Aspergillus flavus Link Resistance of aflatoxin B1 production by Aspergillus flavus Link its inoculation onto four peanuts genotypes

    Directory of Open Access Journals (Sweden)

    Guilherme PRADO

    1999-01-01

    Full Text Available Avaliou-se a produção de aflatoxina B1 pelo Aspergillus flavus IMI 190443, inoculado em sementes de genótipos de amendoim: Tatu Vermelho, VRR-245, 2117 e 2155, in natura e autoclavado a 1210 C por 20 minutos, previamente cultivados no Centro Experimental do Instituto Agronômico de Campinas, em 1995/96. A aflatoxina B1 foi quantificada por cromatografia em camada delgada, utilizando comparação visual com padrões. Os níveis de aflatoxina B1 das amostras autoclavadas e in natura dos genótipos Tatu Vermelho, VRR-245 e 2155 foram significativamente diferentes (p The levels of aflatoxin B1 in postharvest peanuts were evaluated after inoculation of seeds of four different genotypes : Tatu Vermelho, VRR-245, 2117 e 2155 with Aspergillus flavus IMI 190443. Field trials were conducted in 1995/96 at the Experimenal Center of the Instituto Agronômico de Campinas. The postharvest seeds were used in natura or autoclaved at 121 oC for 20 minutes prior to inoculation with the mold. The aflatoxin B1 was quantified by thin layer chromatography (TLC using a visual comparation with standards. The levels of aflatoxin B1 in autoclaved and in natura seeds were significantly different (p < 0,05 between the genotypes Tatu Vermelho, VRR-245, 2155 and the 2117. Samples of genotypes 2117, originating from India, presented the lowest levels of aflatoxin B1 irrespective of heating treatment or not, when compared with the others. The results show possibilities of doing explorations about the varietal resistance in the aflatoxin control.

  7. La exposición a la aflatoxina B1 en animales de laboratorio y su significado en la salud pública Exposure to aflatoxin B1 in experimental animals and its public health significance

    Directory of Open Access Journals (Sweden)

    Doralinda Guzmán de Peña

    2007-06-01

    Full Text Available En México se ha detectado la presencia AFB1 en humanos: como mutación en el gene p53 en hepatocarcinomas de pacientes de Monterrey, Nuevo León, México, en 1996 y como aducto AFB1-lisina en suero de pacientes del Instituto Mexicano del Seguro Social de Matamoros, Tamaulipas, México, en 2003. La aflatoxina B1 ha sido clasificada por la Agencia Internacional para Investigación en Cáncer como un agente carcinogénico para humanos. Este compuesto es un contaminante natural encontrado en alimentos y es sintetizado por Aspergillus flavus y/o A. parasiticus cuando estos hongos crecen en diversos productos alimenticios. Considerando el riesgo que este compuesto representa para los seres humanos, en el presente artículo se revisa y analiza, a nivel molecular, su capacidad carcinogénica, mutagénica y tóxica y se ilustra su relación causal con hepatocarcinomas en humanos. Se destaca que la capacidad carcinogénica y mutagénica están determinadas por la AFB1-formamidopirimidina, la cual causa errores en las transcripciones del ADN. Los resultados ilustran que la población mexicana está consumiendo alimentos con bajas concentraciones de AFB1. La toxicidad es consecuencia de la acción carcinogénica en el hígado.The presence of AFB1 in human beings was detected in Mexico in 1996 both as a mutation of the gene p53 in hepatocellular carcinomas in Monterrey, Mexico, and as the adduct AFB1-lysine in serum from patients in Matamoros, Mexico in 2003. Aflatoxin B1 has been classified as a carcinogenic agent to humans by the International Agency for Research on Cancer. The compound is a natural contaminant produced by Aspergillus flavus and/or A. parasiticus when these fungi grow on different food products. At the molecular level, this review covers the carcinogenic, mutagenic and toxic properties of these mycotoxins and their risk to humans. It also gives insight into the causal relationship between aflatoxins and hepatocellular carcinoma

  8. Determination of Aflatoxin B1 in Raw Materials of Beer by SPE-HPLC%固相萃取-高效液相色谱法测定啤酒原辅料中黄曲霉毒素B1

    Institute of Scientific and Technical Information of China (English)

    杜元正; 蔡国林; 高献礼; 陆健

    2012-01-01

    By comparing the recoveries of different solid phase extraction columns for aflatoxin Bl ( AFB1 ) clean- up, the silicone-diatomite-neutral alumina mixed column was chosen, and the analytical method of Reversed Phase High Performance Liquid Chromatography (RP-HPLC) was developed for determination of aflatoxin B1 in raw materi- als of beer (barley malt, barley, rice and wheat malt). The results showed that the linear range of detection of AFBt ranged from 0.5μg/kg to 50 μg/kg. Standard curve correlation coefficient(r) was 0. 9997. The detection limit was 0. 15μg/kg. The average recovery ranged from 80. 1% to 109.2%, and the relative standard deviation(RSD) ranged from 0.23% to 4.29%. The method was simple, accurate and cost saving. It was also proposed to be used for the quality control of AFB1 in raw materials of beer.%以啤酒酿造中主要原辅料(大麦麦芽、大麦、大米、小麦麦芽)为样品,比较了几种不同净化柱净化黄曲霉毒素B1(AFB1)的回收率,最终选择了硅胶-硅藻土-中性氧化铝混合柱作为净化柱,建立了一种检测啤酒原辅料中AFB。的反相高效液相色谱(RP—HPLC)方法。结果表明,AFB1的检测线性范围为0.5—50μg/kg,线性相关系数r为0.9997。方法检出限为0.15μg/kg,加标回收率80.1%~109.2%,相对标准偏差(RSD)为0.23%-4.29%。该方法简便、准确、成本低廉,可用于啤酒原辅料中的AFB1的质量控制。

  9. Biodegradation of aflatoxin B1 in contaminated rice straw by Pleurotus ostreatus MTCC 142 and Pleurotus ostreatus GHBBF10 in the presence of metal salts and surfactants.

    Science.gov (United States)

    Das, Arijit; Bhattacharya, Sourav; Palaniswamy, Muthusamy; Angayarkanni, Jayaraman

    2014-08-01

    Aflatoxin B1 (AFB1) is a highly toxic fungal metabolite having carcinogenic, mutagenic and teratogenic effects on human and animal health. Accidental feeding of aflatoxin-contaminated rice straw may be detrimental for ruminant livestock and can lead to transmission of this toxin or its metabolites into the milk of dairy cattle. White-rot basidiomycetous fungus Pleurotus ostreatus produces ligninolytic enzymes like laccase and manganese peroxidase (MnP). These extracellular enzymes have been reported to degrade many environmentally hazardous compounds. The present study examines the ability of P. ostreatus strains to degrade AFB1 in rice straw in the presence of metal salts and surfactants. Laccase and MnP activities were determined spectrophotometrically. The efficiency of AFB1 degradation was evaluated by high performance liquid chromatography. Highest degradation was recorded for both P. ostreatus MTCC 142 (89.14 %) and P. ostreatus GHBBF10 (91.76 %) at 0.5 µg mL(-1) initial concentration of AFB1. Enhanced degradation was noted for P. ostreatus MTCC 142 in the presence of Cu(2+) and Triton X-100, at toxin concentration of 5 µg mL(-1). P. ostreatus GHBBF10 showed highest degradation in the presence of Zn(2+) and Tween 80. Liquid chromatography-mass spectrometric analysis revealed the formation of hydrated, decarbonylated and O-dealkylated products. The present findings suggested that supplementation of AFB1-contaminated rice straw by certain metal salts and surfactants can improve the enzymatic degradation of this mycotoxin by P. ostreatus strains. PMID:24770873

  10. Glutathione-S-transferase omega 1 (GSTO1-1) acts as mediator of signaling pathways involved in aflatoxin B1-induced apoptosis-autophagy crosstalk in macrophages.

    Science.gov (United States)

    Paul, Souren; Jakhar, Rekha; Bhardwaj, Monika; Kang, Sun Chul

    2015-12-01

    Aflatoxin B1 (AFB1) is the most toxic aflatoxin species and has been shown to be associated with specific as well as non-specific immune responses. In the present study, using murine macrophage Raw 264.7 cells as a model, we report that short exposure (6h) to AFB1 caused an increase in the cellular calcium pool in mitochondria, which in turn elevated reactive oxygen species (ROS)-mediated oxidative stress and led to loss of mitochondrial membrane potential and ultimately c-Jun N-terminal kinases (JNK)-mediated caspase-dependent cell death. On the contrary, longer exposure (12h) to AFB1 reduced JNK phosphorylation and cell death in macrophages. Measurement of autophagic flux demonstrated that autophagy induction through the canonical pathway was responsible for suppressing AFB1-induced apoptosis after 12h. As a detailed molecular mechanism, we found that the unfolded protein response (UPR) machinery was active at 12h post-exposure to AFB1 and induced cytoprotective autophagy as confirmed by determination of major autophagic markers. Inhibition of autophagy by Beclin-1 siRNA also resulted in JNK-mediated cell death. We further established that glutathione S transferase omega1-1 (GSTO1-1), a specific class of GST, was the responsible factor between apoptosis and autophagy crosstalk. Targeting of GSTO1-1 increased JNK-mediated apoptosis by 2-fold compared to the control, whereas autophagy rate was reduced. Thus, increased expression of GSTO1-1 was associated with increased protein glutathionylation, an important protein modification in response to cellular redox status.

  11. HPLC法测定150种中药材中黄曲霉毒素G2、G1、B2、B1的含量%Determination of Aflatoxin G2,G1,B2 and B1 in 150 Chinese Herbs by HPLC

    Institute of Scientific and Technical Information of China (English)

    郭巧技; 高咏莉; 王淑红

    2012-01-01

    Objective: To establish an HPLC method for the determination of aflatoxin G2 , G1 , B2 and B1 in 150 Chinese herbs. Method: After extracted by 70% methanol and purified by immunoafinity column, the aflatoxins were determined by fluorescence detection. Result: The calibration curves for aflatoxin G2 and B2 were linear within the range of 0. 15-6.00 ng · ml-1 , and those for aflatoxin G1 and B1 were linear within the range of 0. 5-20.00 ng · ml-1 . The recoveries were within the range of 85. 6% -92. 0% . Conclusion : The method is reliable, accurate and specific, and can be used in the determination of aflatoxin G2 , G1 , B2 and B1.%目的:建立了HPLC法测定150种中药材中的黄曲霉毒素G2、G1、B2、B1含量.方法:样品经70% 甲醇提取、免疫亲和色谱柱净化后,用HPLC-柱后衍生-荧光检测器测定.结果:黄曲霉毒素G2、B2在0.15~6.00 ng·ml-1范围内,黄曲霉毒素G1、B1在0.5~20.00 ng·ml-1范围内线性关系良好.回收率为85.6%~92.0%.结论:本法操作简便,结果准确、重复性好,可用于中药材中黄曲霉毒素G2、G1、B2、B1的测定.

  12. Effects of Banzhilian Extract on Notch1 of Aflatoxin B1-induced Hepatic Carcinoma%半枝莲提取物对黄曲霉素 B1所诱发肝癌 Notch1的影响

    Institute of Scientific and Technical Information of China (English)

    蔡林雪; 刘澍楠

    2015-01-01

    Objective:To investigate the Banzhilian extract’s effects on Notch1 of rats with aflatoxin B1-induced hepatic carcino-ma.Methods:Sixty Wistar rats were randomly divided into 4 groups:normal control group,hepatocarcinoma model group,extract’ s high dose group and low dose group.Hepatic carcinoma model were induced by aflatoxin B1.The control group and the model group were given distilled water lavagely, the extract ’s groups were given different concentrations of extracts lavagely, qd.for a month.All rats were killed in the 4th week,measure weight,counted number of nodules on the surface of liver.Immunohistochemis-try were used to detect the expression of Notch1.Results:Compared with the control group,the rats’weight in other groups dropped significantly ( P<0.05) ,while the nodule number and Notch1 levels increased significantly ( P<0.05);Compared with the mod-el group,the rats’weight in the extract’s of high dose group and low dose group were increased ( P<0.05) ,while the nodule num-ber and Notch1 levels were reduced considerably(P<0.05).Conclusion:Banzhilian extract could inhibit aggravation of liver car-cinoma,which is probably related to the down-regulating of liver Notch1.%目的:探讨半枝莲提取物( ESB)对黄曲霉素B1所诱发的肝癌大鼠Notch1的影响。方法:选取60只Wistar大鼠,随机分为空白组、模型组、半枝莲提取物低、高剂量组,每组15只。建立用黄曲霉素B1所诱发的肝癌大鼠的动物模型。其中空白组与模型组给予蒸馏水灌胃,半枝莲提取物低、高剂量组给予不同浓度的ESB灌胃治疗。灌胃1次/d,连续治疗1个月。治疗结束后将所有大鼠处死,观察各组大鼠的体质量、肝表面的结节数,并取肝组织采用免疫组化法检测肝组织中Notch1的表达情况。结果:与空白组比较,模型组和ESB低、高剂量组大鼠的体质量明显降低( P<0.05),肝表面的结节数明显增多(P<0.05

  13. Decontamination of Aflatoxin B1 with Electrolysed Oxidising Water and Security Evaluation%氧化电解水清除黄曲霉毒素B1生成产物及其安全性评价

    Institute of Scientific and Technical Information of China (English)

    熊科; 王晓玲; 李秀婷; 孙宝国; 刘海杰

    2014-01-01

    真菌毒素是引发食品安全问题的重要污染原因.黄曲霉毒素B1(AFB1)因污染范围广,对人和各种生物毒性极强而引起广泛关注.本文利用酸性氧化电解水清除AFB1,对生成产物进行辨识和安全性评价.揭示电酸性氧化电解水清除AFB1的机制.试验结果表明:经高分辨质谱分析,酸性氧化电解水清除AFB1生成的主要产物相对分子质量为364,化学式为C17H13Cl07.结合NMR分析,确认未知产物为8-氯-9-羟基-Aflatoxin B1(8-Cl-9-OH-AFB1).它是具有两亲性质的有机分子.该产物的安全性研究结果表明:产物无致突变性,其半数致死剂量浓度IC50值约为150 mmol/L,对细胞无毒害作用.本文明确了酸.性水清除AFB1的应用安全性,为清除AFB1污染提供了1种安全有效,具有较好应用前景的新途径.

  14. 食用油中黄曲霉毒素B1高效低耗检测技术研究%High efficiency and low-cost technology for determining aflatoxin B1 in edible oil

    Institute of Scientific and Technical Information of China (English)

    刘旭; 张雪梅; 党献民; 任正东; 李尧; 龚珊; 呼鑫

    2012-01-01

    A high efficient and low - cost detection method was developed by the research on the currently used detection methods of aflatoxin B, (AFT B, ). AFT B, was extracted from edible oil with methanol -water (7:3) , and the extract was analyzed by high performance liquid ehromatography (HPLC ) coupled with post column derivatization system and fluorescence detection (FLD) after freeze -filtering. The detection limit was 0.4 μg/kg, relative standard deviation was 1. 25% -2.42% and standard recovery was 93.47% -96.78%. Compared with other instrument detection methods, this method had no clean -up column, and made the detection time shorter and the cost lower.%通过对目前常用的黄曲霉毒素B1(AFTB1)检测方法的研究,开发出一种高效、低耗的检测方法.该方法采用甲醇-水(7∶3)溶液提取食用油中的黄曲霉毒素B1,提取液经冷冻过滤后,通过配有柱后衍生系统和荧光检测器(FLD)的高效液相色谱(HPLC)分析.该方法最低检出限为0.4μg/kg,相对标准偏差为1.25% ~2.42%,加标回收率为93.47% ~96.78%.该方法与其他仪器检测方法相比不用任何净化小柱,缩短了检测时间,降低了检测成本.

  15. An investigation of aflatoxin B1 in edible oil in 2011 and 2012%2011-2012年食用植物油中黄曲霉毒素B1的调查

    Institute of Scientific and Technical Information of China (English)

    刘晓莉; 曹悦; 陈世琼; 蔡雪凤; 曹宝森

    2012-01-01

    To survey how did the Aflatoxin BI in edible oil meet the standards, one thousand three hundred and thirty edible oil samples ( including 988 traditional cooking oil and 342 health function and seasoning oil samples ) were detected by ELISA method from 2011 to 2012. The results showed that in 2011, the pass rate of traditional cooking oil was 99.22%, and of the health function oil and seasoning oil was 95.03%. In 2012, the pass rate of traditional cooking oil was 99.37%, and of the health function oil and seasoning oil was 96.90%. It was indicated that the quality of edible oil was improving year by year,%为调查我国市场食用植物油油中黄曲礞毒素B1是螽符合国家标准规定,本论义膈两年的时间,对988个传统烹调油样品、342个新兴功放类油样品中的黄曲霉毒素B1进行了检测。结果显示:2011年传统烹调油合格率为99.22%,新必功效类油样,铺及凋味汕合格宰为95.03%;2012年传统烹调油合格率为99.37%,新必功效类油样品及调味油合格半为96.90%。由此可见,食用油产品的合格牢征逐年提高。

  16. cDNA cloning, expression and activity of a second human aflatoxin B1-metabolizing member of the aldo-keto reductase superfamily, AKR7A3.

    Science.gov (United States)

    Knight, L P; Primiano, T; Groopman, J D; Kensler, T W; Sutter, T R

    1999-07-01

    The aflatoxin B1 (AFB1) aldehyde metabolite of AFB1 may contribute to the cytotoxicity of this hepatocarcinogen via protein adduction. Aflatoxin B1 aldehyde reductases, specifically the NADPH-dependent aldo-keto reductases of rat (AKR7A1) and human (AKR7A2), are known to metabolize the AFB1 dihydrodiol by forming AFB1 dialcohol. Using a rat AKR7A1 cDNA, we isolated and characterized a distinct aldo-keto reductase (AKR7A3) from an adult human liver cDNA library. The deduced amino acid sequence of AKR7A3 shares 80 and 88% identity with rat AKR7A1 and human AKR7A2, respectively. Recombinant rat AKR7A1 and human AKR7A3 were expressed and purified from Escherichia coli as hexa-histidine tagged fusion proteins. These proteins catalyzed the reduction of several model carbonyl-containing substrates. The NADPH-dependent formation of AFB1 dialcohol by recombinant human AKR7A3 was confirmed by liquid chromatography coupled to electrospray ionization mass spectrometry. Rabbit polyclonal antibodies produced using recombinant rat AKR7A1 protein were shown to detect nanogram amounts of rat and human AKR7A protein. The amount of AKR7A-related protein in hepatic cytosols of 1, 2-dithiole-3-thione-treated rats was 18-fold greater than in cytosols from untreated animals. These antibodies detected AKR7A-related protein in normal human liver samples ranging from 0.3 to 0.8 microg/mg cytosolic protein. Northern blot analysis showed varying levels of expression of AKR7A RNA in human liver and in several extrahepatic tissues, with relatively high levels in the stomach, pancreas, kidney and liver. Based on the kinetic parameters determined using recombinant human AKR7A3 and AFB1 dihydrodiol at pH 7.4, the catalytic efficiency of this reaction (k2/K, per M/s) equals or exceeds those reported for other enzymes, for example cytochrome P450s and glutathione S-transferases, known to metabolize AFB1 in vivo. These findings indicate that, depending on the extent of AFB1 dihydrodiol formation, AKR

  17. Quantitative analysis of Aflatoxin B1 using the microsphere array technology%霉毒素B1液相芯片定量检测方法的探索

    Institute of Scientific and Technical Information of China (English)

    宋慧君; 马惠蕊; 裴轶君; 刘淑艳; 曹远银

    2011-01-01

    采用间接竞争法的原理和液相芯片技术平台,选用黄曲霉毒素B1( AFB1)多克隆抗体对AFB1的定量检测方法进行了探索.通过优化偶联抗原浓度,确定AFB1多抗临界饱和浓度和抗原抗体最佳孵育时间,建立了AFB1液相芯片定量检测方法.以通用的IC5o值作为灵敏度衡量标准,其灵敏度为1.33 ng/mL,以IC1o作为最低检测限衡量标准,其最低检测限为0.15 ng/mL,线性方程为y=0.000 6x +0.000 8.应用该方法对脱脂牛奶和全脂牛奶中的AFB1进行添加回收率检测,在2.0~16.0 ng/mL添加水平下,平均回收率均大于75%,相对标准偏差介于2.80% ~4.69%(n=7)之间.经过实际样品的检测,证明该方法灵敏、稳定、快速、简便,适用于大量样本的检测.%The quantitative detection of Aflatoxin Bl (AFB1) was investigated on the microsphere array platform and indirect competition by using the Anti-Aflatoxin Bl. Through optimizing the concentration and saturated concentration of AFB1-BSA and Anti-Aflatoxin Bl as well as incubation time of antigen and antibody, the method of quantitative detection of AFB1 was established. As referring to the detection sensitivity standard IC50, results showed that the detection sensitivity was 1.33 ng/mL. Meanwhile,as referring to the limit of detection standard IC]0 ,the detection limition was 0.15 ng/mL. The linear equation was v =0. 000 6x + 0. 000 8. By applying this original research to test the imitating contaminated skim and full milk,the recovery rate of the two kinds of milk was all greater than 75% under the AFB1 added level of 2. 0 - 16. 0 ng/mL, and the relative standard deviation was between 2. 80% -4. 69% (n =7). Overall, the quantitative analysis of AFB1 using the microsphere arraytechnology was sensitive,stable,rapid and simple. The method was applicable to mass sample testing.

  18. 小型花生榨油厂对黄曲霉毒素B_1防控与技改措施%The Prevention and Technical Innovation Measures to the Aflatoxin B_1 in Small Peanut Oil Extraction Factory

    Institute of Scientific and Technical Information of China (English)

    何春林; 张庆珍

    2012-01-01

    The aflatoxin was produced by Aspergillus flavus.The peanuts have a strong carcinogenic effect to humans and animals after being contaminated.The current pollution of aflatoxin B1 in peanut oil were described,the reasons to,prevention and innovation measures were elaborated,the focus on the small peanut oil extraction factory in raw materials and processing technology control was discussed,the aim was to play a guidance role in the prevention to the aflatoxin B1 and keep the amount of aflatoxin B1 in a security level.%花生被黄曲霉菌污染后产生的黄曲霉毒素对人和动物有很强的致癌作用。叙述了目前花生油中黄曲霉毒素B1的污染状况,对引起黄曲霉毒素B1产生的原因、防控方法和技改措施等方面进行了阐述,重点对小型花生榨油厂在原料控制和加工过程中技术控制上进行探讨,旨在对小型花生榨油厂在黄曲霉毒素B1的防控上起到指导作用,将黄曲霉毒素B1量控制在安全水平。

  19. Quality control charts used in the determination of aflatoxin B1 in rice by HPLC with post-column derivatization%HPLC柱后衍生法测定黄曲霉毒素 B1质量控制图的应用

    Institute of Scientific and Technical Information of China (English)

    李涛; 周艳华; 龙凌云; 潘小红

    2016-01-01

    研究高效液相色谱柱后衍生法测定黄曲霉毒素B1的质量控制图的应用。通过加标样品的测试,用数理统计的方法求出相关技术指标,作出黄曲霉毒素B1质量控制图,得出A FB1的控制限为73.78%~90.24%,警戒线为76.52%~87.50%。试验结果表明,质量控制图能直观有效地控制高效液相色谱柱后衍生法测定黄曲霉毒素B1的分析质量,确保检测结果准确可靠。%ABSTRACT:The quality control methods in the determination of aflatoxin B1 by HPLC with post‐column derivatization were studied .The recovery of standard addition were tested and the relevant technical indexes were worked out using mathematical statistics to prepare the quality control charts of the determination of aflatoxin B1 .The control limit of aflatoxin B1 was 73 .78% ~90 .24% and the warning limit was 76 .52% ~87 .50% .The quality control charts could directly and effectively contol the quality of the determination of aflatoxin B1 in rice and ensure accurate and reliable results .

  20. Simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2 in peanuts and their derivative products by ultra-high-performance liquid chromatography-tandem mass spectrometry.

    Science.gov (United States)

    Huang, Baifen; Han, Zheng; Cai, Zengxuan; Wu, Yongjiang; Ren, Yiping

    2010-03-01

    A reliable ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for simultaneous determination of aflatoxins B1, B2, G1, G2, M1 and M2 in peanuts and their derivative products was developed. The sample was extracted by 84% of acetonitrile aqueous solution and the extract was purified by a reliable solid phase extraction-based clean-up method. Then, the analytes were separated on Acquity UPLC HSS T3 column (100 mm x 2.1 mm, 1.8 microm particle size), and eluted with a mobile phase consisting of (A) water containing 0.1% formic acid and (B) acetonitrile/methanol (50/50, v/v). The separated compounds were detected with a Waters Micromass Quattro Ultima Pt tandem quadrupole mass spectrometer operating in positive electro-spray ionization using multiple reaction monitoring mode. The established method was extensively validated by determining the linearity (R(2) > or = 0.9990), average recovery (74.7-86.8%) and precision (relative standard deviation aflatoxins in peanuts and their derivative products. Finally, a total of 73 samples randomly collected from different areas in Zhejiang province were screened for aflatoxins with the proposed method. The results showed that 31 samples of peanut butter, 14 samples of fresh peanut and 5 samples of musty peanut were contaminated with aflatoxins. Meanwhile, this was the first report on aflatoxins M1 and M2, which were found in unprocessed peanuts and their derivative products. PMID:20152266

  1. Survey of Deoxynivalenol and Aflatoxin B1 in Instant Noodles and Bread Consumed in Thailand by Using Liquid Chromatography-Tandem Mass Spectrometry.

    Science.gov (United States)

    Pralatnet, Sasithorn; Poapolathep, Saranya; Giorgi, Mario; Imsilp, Kanjana; Kumagai, Susumu; Poapolathep, Amnart

    2016-07-01

    One hundred wheat product samples (50 instant noodle samples and 50 bread samples) were collected from supermarkets in Bangkok, Thailand. Deoxynivalenol (DON) and aflatoxin B1 (AFB1) contamination in these products was analyzed using a validated liquid chromatography-tandem mass spectrometry method. The limit of quantification values of DON and AFB1 in the instant noodles and bread were 2 and 1 ng g(-1), respectively. The survey found that DON was quantifiable in 40% of collected samples, in 2% of noodles (0.089 μg g(-1)), and in 78% of breads (0.004 to 0.331 μg g(-1)). AFB1 was below the limit of quantification of the method in all of the tested samples. The results suggest that the risk of DON exposure via noodles and breads is very low in urban areas of Thailand. No risk can be attributable to AFB1 exposure in the same food matrices, but further studies with a larger sample size are needed to confirm these data.

  2. Performance improvement of the one-dot lateral flow immunoassay for aflatoxin B1 by using a smartphone-based reading system.

    Science.gov (United States)

    Lee, Sangdae; Kim, Giyoung; Moon, Jihea

    2013-01-01

    This study was conducted to develop a simple, rapid, and accurate lateral flow immunoassay (LFIA) detection method for point-of-care diagnosis. The one-dot LFIA for aflatoxin B1 (AFB1) was based on the modified competitive binding format using competition between AFB1 and colloidal gold-AFB1-BSA conjugate for antibody binding sites in the test zone. A Smartphone-based reading system consisting of a Samsung Galaxy S2 Smartphone, a LFIA reader, and a Smartphone application for the image acquisition and data analysis. The detection limit of one-dot LFIA for AFB1 is 5 μg/kg. This method provided semi-quantitative analysis of AFB1 samples in the range of 5 to 1,000 μg/kg. Using combination of the one-dot LFIA and the Smartphone-based reading system, it is possible to conduct a more fast and accurate point-of-care diagnosis. PMID:23598499

  3. Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1

    Science.gov (United States)

    Farzaneh, Mohsen; Shi, Zhi-Qi; Ahmadzadeh, Masoud; Hu, Liang-Bin; Ghassempour, Alireza

    2016-01-01

    In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1), caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC) method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut. PMID:27298596

  4. Inhibition of the Aspergillus flavus Growth and Aflatoxin B1 Contamination on Pistachio Nut by Fengycin and Surfactin-Producing Bacillus subtilis UTBSP1

    Directory of Open Access Journals (Sweden)

    Mohsen Farzaneh

    2016-06-01

    Full Text Available In this study, the treatment of pistachio nuts by Bacillus subtilis UTBSP1, a promising isolate to degrade aflatoxin B1 (AFB1, caused to reduce the growth of Aspergillus flavus R5 and AFB1 content on pistachio nuts. Fluorescence probes revealed that the cell free supernatant fluid from UTBSP1 affects spore viability considerably. Using high-performance liquid chromatographic (HPLC method, 10 fractions were separated and collected from methanol extract of cell free supernatant fluid. Two fractions showed inhibition zones against A. flavus. Mass spectrometric analysis of the both antifungal fractions revealed a high similarity between these anti-A. flavus compounds and cyclic-lipopeptides of surfactin, and fengycin families. Coproduction of surfactin and fengycin acted in a synergistic manner and consequently caused a strong antifungal activity against A. flavus R5. There was a positive significant correlation between the reduction of A. flavus growth and the reduction of AFB1 contamination on pistachio nut by UTBSP1. The results indicated that fengycin and surfactin-producing B. subtilis UTBSP1 can potentially reduce A. flavus growth and AFB1 content in pistachio nut.

  5. Label-free immunosensor based on one-step electrodeposition of chitosan-gold nanoparticles biocompatible film on Au microelectrode for determination of aflatoxin B1 in maize.

    Science.gov (United States)

    Ma, Haihua; Sun, Jizhou; Zhang, Yuan; Bian, Chao; Xia, Shanhong; Zhen, Tong

    2016-06-15

    Gold nanoparticles (AuNPs) embedded in chitosan (CHI) film, well-dispersed and smaller in size (about 10 nm), were fabricated by one-step electrodeposion on Au microelectrode in solution containing chitosan and chloride trihydrate. The nano-structure CHI-AuNPs composite film offers abundant amine groups, good conductivity, excellent biocompatibility and stability for antibody immobilization. The combination of aflatoxin B1 (AFB1) with immobilized antibody introduces a barrier to electron transfer, resulting in current decreasement. The morphologies and characterizations of modified microelectrodes were investigated by scanning electron microscope (SEM), cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and Fourier transform infrared spectroscopy (FT-IR). The proposed non-enzyme and label-free immunosensor exhibited high sensitive amperometric response to AFB1 concentration in two linear ranges of 0.1 to 1 ng mL(-1) and 1 to 30 ng mL(-1), with the detection limit of 0.06 ng mL(-1) (S/N=3). The immunoassay was also applied for analysis of maize samples spiked with AFB1. Considering the sample extraction procedure, the linear range and limit of detection were assessed to be 1.6-16 ng mL(-1) and 0.19 ng mL(-1) respectively. The simple method showed good fabrication controllability and reproducibility for immunosensor design. PMID:26851579

  6. Toxic effects of aflatoxin B1 on embryonic development of zebrafish (Danio rerio): potential activity of piceatannol encapsulated chitosan/poly (lactic acid) nanoparticles.

    Science.gov (United States)

    Dhanapal, Jeevitha; Ravindrran, Malathy Balaraman; Baskar, Santhosh K

    2015-01-01

    The aim was to analyse the efficacy of piceatannol (PIC) loaded chitosan (CS)/poly(lactic acid)(PLA) nanoparticles (CS/PLA-PIC NPs) in zebra fish embryos exposed to aflatoxin B1 (AFB1). FTIR confirmed the chemical interaction between the polymers and drug. SEM showed the size of CS/PLA-PIC NPs approximately 87 to 200nm, compared to CS-PLA NPs of 150nm size. The size was further affirmed as 127nm (CS-PLA NPs) and 147nm (CS/PLA-PIC NPs) by zetasizer depiction. CS/PLA-PIC NPs have not illustrated toxicity at high concentrations when tested in zebrafish embryos. AFB1 wielded their toxic effects on the survival, spontaneous movement, hatching and heart rate and development of embryos were observed in both time and dose-dependent manner at 4μM. Our results suggested that the addition of CS/PLA-PIC NPs increases the survival, heart rate and hatching in time dependent manner at the dosage of 20μg/ml. These hopeful results may prompt the advancement of drug encapsulated polymeric nanoparticles which may have the potential role in improving the AFB1 induced toxicity in humans as well. PMID:25322988

  7. Hetero-enzyme-based two-round signal amplification strategy for trace detection of aflatoxin B1 using an electrochemical aptasensor.

    Science.gov (United States)

    Zheng, Wanli; Teng, Jun; Cheng, Lin; Ye, Yingwang; Pan, Daodong; Wu, Jingjing; Xue, Feng; Liu, Guodong; Chen, Wei

    2016-06-15

    An electrochemical aptasensor for trace detection of aflatoxin B1 (AFB1) was developed by using an aptamer as the recognition unit while adopting the telomerase and EXO III based two-round signal amplification strategy as the signal enhancement units. The telomerase amplification was used to elongate the ssDNA probes on the surface of gold nanoparticles, by which the signal response range of the signal-off model electrochemical aptasensor could be correspondingly enlarged. Then, the EXO III amplification was used to hydrolyze the 3'-end of the dsDNA after the recognition of target AFB1, which caused the release of bounded AFB1 into the sensing system, where it participated in the next recognition-sensing cycle. With this two-round signal amplified electrochemical aptasensor, target AFB1 was successfully measured at trace concentrations with excellent detection limit of 0.6*10(-4)ppt and satisfied specificity due to the excellent affinity of the aptamer against AFB1. Based on this designed two-round signal amplification strategy, both the sensing range and detection limit were greatly improved. This proposed ultrasensitive electrochemical aptasensor method was also validated by comparison with the classic instrumental methods. Importantly, this hetero-enzyme based two-round signal amplified electrochemical aptasensor offers a great promising protocol for ultrasensitive detection of AFB1 and other mycotoxins by replacing the core recognition sequence of the aptamer. PMID:26896792

  8. Phlomis mauritanica extracts reduce the xanthine oxidase activity, scavenge the superoxide anions, and inhibit the aflatoxin B1-, sodium azide-, and 4-nitrophenyldiamine-induced mutagenicity in bacteria.

    Science.gov (United States)

    Limem, Ilef; Bouhlel, Ines; Bouchemi, Meriem; Kilani, Soumaya; Boubaker, Jihed; Ben-Sghaier, Mohamed; Skandrani, Ines; Behouri, Wissem; Neffati, Aicha; Ghedira, Kamel; Chekir-Ghedira, Leila

    2010-06-01

    Four extracts were prepared from the leaves of Phlomis mauritanica: lyophilized infusion, total oligomer flavonoids, methanol, and ethyl acetate extracts. The antimutagenic properties of these extracts were investigated by assessing the inhibition of the mutagenic effects of direct-acting mutagens such as sodium azide and 4-nitrophenylenediamine and indirect-acting mutagens like aflatoxin B1 (AFB1) using the Ames assay. The four extracts prepared from P. mauritanica strongly inhibit the mutagenicity induced by AFB1 in both Salmonella typhimurium TA 100 and TA 98 assay systems. Lyophilized infusion and methanol extracts at the dose of 250 microg per plate reduced AFB1 mutagenicity by 93% and 91%, respectively, in S. typhymurium strain TA 100. We examined also the antioxidant effect of these extracts by the enzymatic xanthine/xanthine oxidase assay. Result indicated that total oligomer flavonoids and ethyl acetate and methanol extracts were potent inhibitors of xanthine oxidase activity. In contrast, lyophilized infusion, total oligomer flavonoids, and methanol extracts exhibited a high degree of superoxide anion scavenging. Our findings emphasize the potential of P. mauritanica extracts to prevent mutations and oxidant effects. Furthermore, the results presented here could be an additional argument to support the use of this species as a medicinal and dietary plant. PMID:20406134

  9. Performance Improvement of the One-Dot Lateral Flow Immunoassay for Aflatoxin B1 by Using a Smartphone-Based Reading System

    Directory of Open Access Journals (Sweden)

    Jihea Moon

    2013-04-01

    Full Text Available This study was conducted to develop a simple, rapid, and accurate lateral flow immunoassay (LFIA detection method for point-of-care diagnosis. The one-dot LFIA for aflatoxin B1 (AFB1 was based on the modified competitive binding format using competition between AFB1 and colloidal gold-AFB1-BSA conjugate for antibody binding sites in the test zone. A Smartphone-based reading system consisting of a Samsung Galaxy S2 Smartphone, a LFIA reader, and a Smartphone application for the image acquisition and data analysis. The detection limit of one-dot LFIA for AFB1 is 5 μg/kg. This method provided semi-quantitative analysis of AFB1 samples in the range of 5 to 1,000 μg/kg. Using combination of the one-dot LFIA and the Smartphone-based reading system, it is possible to conduct a more fast and accurate point-of-care diagnosis.

  10. Disposable competitive-type immunoassay for determination of aflatoxin B1 via detection of copper ions released from Cu-apatite.

    Science.gov (United States)

    Wang, Huan; Zhang, Yihe; Chu, Yanguang; Ma, Hongmin; Li, Yan; Wu, Dan; Du, Bin; Wei, Qin

    2016-01-15

    A disposable electrochemical immunosensor was developed for detection of aflatoxin B1 (AFB1) based on stripping voltammetric detection of copper ions released from Cu-apatite. AFB1 antibody (Ab) was firstly fixed on the gold nanoparticle (Au NPs) modified screen-printed carbon electrode (SPCE). AFB1-bovine serum albumin (AFB1-BSA) conjugate was labeled with Cu-apatite, and then competed with AFB1 for binding to the Ab. Copper ions were released from Cu-apatite through acidolysis and stripping voltammetry signal of the copper ions was used for the detection. The Cu-apatite increased the amount of loaded copper ions, and the anodic stripping strategy performed in the micro electrolytic cell of the SPCE simplified the detection procedure and further amplified the electrochemical signal. This immunosensor could detect AFB1 over a wide concentration range from 0.001 to 100ng mL(-1) with a detection limit of 0.2pg mL(-1). The low cost, high sensitive, rapid and accurate method may find widely potential application in the detection of other toxic or harmful substances.

  11. Hetero-enzyme-based two-round signal amplification strategy for trace detection of aflatoxin B1 using an electrochemical aptasensor.

    Science.gov (United States)

    Zheng, Wanli; Teng, Jun; Cheng, Lin; Ye, Yingwang; Pan, Daodong; Wu, Jingjing; Xue, Feng; Liu, Guodong; Chen, Wei

    2016-06-15

    An electrochemical aptasensor for trace detection of aflatoxin B1 (AFB1) was developed by using an aptamer as the recognition unit while adopting the telomerase and EXO III based two-round signal amplification strategy as the signal enhancement units. The telomerase amplification was used to elongate the ssDNA probes on the surface of gold nanoparticles, by which the signal response range of the signal-off model electrochemical aptasensor could be correspondingly enlarged. Then, the EXO III amplification was used to hydrolyze the 3'-end of the dsDNA after the recognition of target AFB1, which caused the release of bounded AFB1 into the sensing system, where it participated in the next recognition-sensing cycle. With this two-round signal amplified electrochemical aptasensor, target AFB1 was successfully measured at trace concentrations with excellent detection limit of 0.6*10(-4)ppt and satisfied specificity due to the excellent affinity of the aptamer against AFB1. Based on this designed two-round signal amplification strategy, both the sensing range and detection limit were greatly improved. This proposed ultrasensitive electrochemical aptasensor method was also validated by comparison with the classic instrumental methods. Importantly, this hetero-enzyme based two-round signal amplified electrochemical aptasensor offers a great promising protocol for ultrasensitive detection of AFB1 and other mycotoxins by replacing the core recognition sequence of the aptamer.

  12. Lateral flow immunodipstick for visual detection of aflatoxin B1 in food using immuno-nanoparticles composed of a silver core and a gold shell

    International Nuclear Information System (INIS)

    An immunodipstick assay with a lateral flow strip was developed for fast screening of food for aflatoxin B1 (AFB1) using the respective monoclonal antibody immobilized on nanoparticles with a silver core and a gold shell (AgAu) as detection reagent. The membrane-based immunodipstick consisted of a test line containing AFB1 conjugated to bovine serum albumin, and a control line with goat anti-mouse IgG. One to two colored lines are formed on the membrane by using the red AgAu nanoparticles coated with anti-AFB1 as signaling reagents. Under optimal conditions, the dipstick exhibits a lower visual detection limit of 0. 1 ng mL-1 of AFB1. Compared to the use of pure gold nanoparticles, the AgAu nanoparticles strongly enhance the sensitivity of the assay, and the reproducibility and stability are comparable. The assay was evaluated with naturally contaminated samples including rice, wheat, sunflower, cotton, chillies, and almonds, and a good correlation was found with data obtained with a commercially available enzyme-linked immunosorbent assay. The simple and non-instrumental dipstick method may further be extended to the screening of other mycotoxins in food. (author)

  13. Indole-3-carbinol induces a rat liver glutathione transferase subunit (Yc2) with high activity toward aflatoxin B1 exo-epoxide. Association with reduced levels of hepatic aflatoxin-DNA adducts in vivo.

    Science.gov (United States)

    Stresser, D M; Williams, D E; McLellan, L I; Harris, T M; Bailey, G S

    1994-01-01

    Aflatoxin B1 (AFB1), a metabolite of the grain mold Aspergillus flavus, is a potent hepatocarcinogen and widespread contaminant of human food supplies. AFB1-induced tumors or preneoplastic lesions in experimental animals can be inhibited by cotreatment with several compounds, including indole-3-carbinol (I3C), a component of cruciferous vegetables, and the well-known Ah receptor agonist beta-naphthoflavone (BNF). This study examines the influence of these two agents on the AFB1-glutathione detoxication pathway and AFB1-DNA adduction in rat liver. After 7 days of feeding approximately equally inhibitory doses of I3C (0.2%) or BNF (0.04%) alone or in combination, male Fischer 344 rats were administered [3H]AFB1 (0.5 mg/kg, 480 microCi/kg) intraperitoneally and killed 2 hr later. All three experimental diets inhibited in vivo AFB1-DNA adduction (BNF, 46%; I3C, 68%; combined, 51%). Based on Western blots using antibodies specific for the glutathione S-transferase (GST), subunit Yc2 (subunit 10) appeared to be substantially elevated by the diets containing I3C (I3C diet, 4.0-fold increase in band density; combined diet, 2.8-fold). The BNF diet appeared to elevate Yc2 to a lesser extent (2.2-fold increase in band density).(ABSTRACT TRUNCATED AT 250 WORDS)

  14. Regulation of rat glutathione S-transferase A5 by cancer chemopreventive agents: mechanisms of inducible resistance to aflatoxin B1.

    Science.gov (United States)

    Hayes, J D; Pulford, D J; Ellis, E M; McLeod, R; James, R F; Seidegård, J; Mosialou, E; Jernström, B; Neal, G E

    1998-04-24

    The rat can be protected against aflatoxin B1 (AFB1) hepatocarcinogenesis by being fed on a diet containing the synthetic antioxidant ethoxyquin. Evidence suggests that chemoprotection against AFB1 is due to increased detoxification of the mycotoxin by one or more inducible drug-metabolising enzymes. The glutathione S-transferase (GST) isoenzymes in rat liver that contribute to ethoxyquin-induced chemoprotection against AFB1 have been identified by protein purification. This approach resulted in the isolation of several heterodimeric class alpha GST, all of which contained the A5 subunit and possessed at least 50-fold greater activity towards AFB1-8,9-epoxide than previously studied transferases. Molecular cloning and heterologous expression of rat GSTA5-5 has led to the demonstration that it exhibits substantially greater activity for AFB1-8,9-epoxide than other rat transferases. The A5 homodimer can also catalyse the conjugation of glutathione with other epoxides, such as trans-stilbene oxide and 1,2-epoxy-3-(4'-nitrophenoxy)propane, and possesses high catalytic activity for the reactive aldehyde 4-hydroxynonenal. Western blotting has shown that the A5 subunit is not only induced by ethoxyquin but that it is also induced by other cancer chemopreventive agents, such as butylated hydroxyanisole, oltipraz, benzyl isothiocyanate, indole-3-carbinol and coumarin. In addition to GSTA5, we have identified a novel aflatoxin-aldehyde reductase (AFAR) that is similarly induced by ethoxyquin. However, immunoblotting has shown that GSTA5 and AFAR are not always co-ordinately regulated by chemoprotectors. In order to gain a better understanding of the mechanisms responsible for the induction of GSTA5 protein, the GSTA5 gene has been cloned. It was isolated on two overlapping bacteriophage lambda clones and found to be approximately 12 kb in length. The transcriptional start site of GSTA5 has been identified 228 bp upstream from the ATG translational initiation codon. Computer

  15. Isolation, screening and identifi cation of Bacillus spp. as direct-fed microbial candidates for aflatoxin B1 biodegradation

    Institute of Scientific and Technical Information of China (English)

    Rosario; Galarza-Seeber; Juan; David; Latorre; Xochitl; Hernandez-Velasco; Amanda; Drake; Wolfenden; Lisa; Renee; Bielke; Anita; Menconi; Billy; Marshall; Hargis; Guillermo; Tellez

    2015-01-01

    Objective: To evaluate the ability of Bacillus spp. as direct-fed microbials(DFM) to biodegrade al atoxin B1(AFB1) by using an in vitro digestive model simulating in vivo conditions.Methods: Sixty-nine Bacillus isolates were obtained from intestines, and soil samples were screened by using a selective media method against 0.25 and 1.00 μg/m L of AFB1 in modii ed Czapek-Dox medium. Plates were incubated at 37 °C and observed every two days for two weeks. Physiological properties of the three Bacillus spp. candidates were characterized biochemically and by 16 S r RNA sequence analyzes for identii cation. Tolerance to acidic p H, osmotic concentrations of Na Cl, bile salts were tested, and antimicrobial sensitivity proi les were also determined. Bacillus candidates were individually sporulated by using a solid fermentation method and combined. Spores were incorporated into 1 of 3 experimental feed groups: 1) Negative control group, with unmedicated starter broiler feed without AFB1; 2) Positive control group, with negative control feed contaminated with 0.01% AFB1; 3) DFM treated group, with positive control feed supplemented with 109 spores/g. After digestion time(3:15 h), supernatants and digesta were collected for high-performance liquid chromatography l uorescence detection analysis by triplicate.Results: Three out of those sixty-nine DFM candidates showed ability to biodegrade AFB1 in vitro based on growth as well as reduction of l uorescence and area of clearance around each colony in modii ed Czapek-Dox medium which was clearly visible under day light after 48 h of evaluation. Analysis of 16S-DNA identii ed the strains as Bacillus amyloliquefaciens, Bacillus megaterium and Bacillus subtilis. The three Bacillus strains were tolerant to acidic conditions(p H 2.0), tolerant to a high osmotic pressure(Na Cl at 6.5%), and were able to tolerate 0.037% bile salts after 24 h of incubation. No signii cant dif erences(P > 0.05) were observed in the concentrations of

  16. Binding Assay for Aflatoxin B_1 by 3 Absorbents%3种吸附剂对黄曲霉毒素B_1吸附能力的研究

    Institute of Scientific and Technical Information of China (English)

    李娟娟; 索德成; 苏晓鸥

    2009-01-01

    [目的]体外试验评价3种吸附剂(吸附剂A:主成分为酵母细胞壁提取物;吸附剂B:主成分为水合铝硅酸盐;吸附剂C为复合物,主成分是酵母细胞壁及水合铝硅酸盐)对黄曲霉毒素B_1(AFB_1)的吸附效果,并通过吸附剂对摄入AFB_1肉仔鸡生长性能及血清蛋白水平的影响验证体外试验结果.[方法](1)pH为2.0、6.0,8.0的磷酸盐缓冲溶液(PBS溶液)、人工胃液和人工肠液环境下吸附剂对AFB_1的吸附能力;(2)吸附剂对AFB_1的结合速率;(3)吸附剂结合AFB_1形成的复合体的稳定性;(4)240只1日龄雄性从肉仔鸡,随机分为8个处理,比较吸附剂对摄入AFB_1肉仔鸡生长性能及血清蛋白水平的影响.[结果](1)5种酸碱条件下吸附剂结合AFB_1的能力大小顺序为B>C>A;(2)吸附剂B在10 min内对AFB_1吸附率达到97.69%,且60 min内一直处在96.03%以上,而吸附剂A、C在60 min内对AFB_1的吸附能力不稳定;(3)与吸附剂A、C相比较,体外条件下吸附剂B与AFB_1形成的复合体解吸附率最低;(4)与基础组比较,AFB_1组肉仔鸡采食量、体重增加显著下降,料重比显著增加(P<0.05),血清总蛋白(TP)、白蛋白(ALB)、球蛋白(GLOB)水平均显著下降(P<0.05).3种吸附剂均能提高肉仔鸡生长性能,吸附剂B能显著改善摄入AFB_1污染日粮(98.98 μg·kg~(-1))的肉仔鸡血清蛋白水平的降低,添加吸附剂A或C效果不显著.[结论] 3种吸附剂对AFB_1均有一定的吸附作用,吸附剂B的作用效果优于吸附剂A和吸附剂C.结果提示,将吸附剂B应用于被AFB_1污染的家禽饲料中,与其它霉菌毒素管理措施相结合,能降低AFB_1对肉仔鸡的危害.%[Objective] In this study, the in vitro sorption of aflatoxin B_1(AFB_1) onto different absorbents was characterized, and the in vitro assay result was verified by comparing growth performance and serum protein levels of broilers exposed to aflatoxin-contamination feed. Three absorbents were product A

  17. HPLC柱后衍生同时测定黄曲霉毒素B1、B2、G1、G2的探讨%Simultaneous determination of aflatoxins B1, B2, G1, G2 by HPLC with postcolumn derivatization

    Institute of Scientific and Technical Information of China (English)

    王新丽; 张玉黔

    2012-01-01

    Objective:To develop a high performance liquid chromatography with postcolumn derivatization method for simultaneous determination of Aflatoxins B1 ,B2,G1 ,G2. Methods:The extracted samples were separated by HPLC column, conducted reaction with iodine solution in postcolumn derivatization device, finally detected by fluorescence detector. Results; The method has a limit of detection of 0.03 |xg/kg for aflatoxins B1 ,G2, 0.01 u,g/kg for aflatoxin B2 and 0.05 ug/kg for aflatoxin G,. Conclusion:This method was sensitive, accurate, simple and specific, which can be applied for the determination of aflatoxins B,, B2,G1 ,G2.%目的:建立黄曲霉毒素B1、B2、G1、G2的高效液相色谱柱后衍生检测方法.方法:样品通过高效液相色谱柱分离后,进入柱后衍生装置与碘溶液发生反应,最后进入荧光检测器进行检测.结果:该方法的检出限AFB1、AFG2为0.03 μg/kg、AFB2为0.01 μg/kg、AFG1为0.05 μg/kg.结论:方法灵敏度高,准确度好,操作简单,实用性强,可用于黄曲霉毒素的检测.

  18. Report on the 2014 Proficiency Test of the European Union Reference Laboratory for Mycotoxins, for the Network of National Reference Laboratories - Determination of Aflatoxin B1 in Copra (Coconut powder)

    OpenAIRE

    KUJAWSKI MACIEJ WOJCIECH; MISCHKE Carsten; Stroka, Joerg

    2014-01-01

    This report presents the results of the PT of the EURL for Mycotoxins which focused on the determination of aflatoxin B1 (Afla B1) in coconut powder samples. Sixty-one participants from 31 countries registered for the exercise and fifty-eight sets of results were reported. Only z-scores were used for an evaluation of performance. In total 91 % of the attributed z scores were below an absolute value of two, which indicated that most of the participants performed satisfactorily.

  19. Sulforaphane- and phenethyl isothiocyanate-induced inhibition of aflatoxin B1-mediated genotoxicity in human hepatocytes: role of GSTM1 genotype and CYP3A4 gene expression.

    Science.gov (United States)

    Gross-Steinmeyer, Kerstin; Stapleton, Patricia L; Tracy, Julia H; Bammler, Theo K; Strom, Stephen C; Eaton, David L

    2010-08-01

    Primary cultures of human hepatocytes were used to investigate whether the dietary isothiocyanates, sulforaphane (SFN), and phenethyl isothiocyanate (PEITC) can reduce DNA adduct formation of the hepatocarcinogen aflatoxin B(1) (AFB). Following 48 h of pretreatment, 10 and 50 microM SFN greatly decreased AFB-DNA adduct levels, whereas 25muM PEITC decreased AFB-DNA adducts in some but not all hepatocyte preparations. Microarray and quantitative reverse transcriptase (RT)-PCR analyses of gene expression in SFN and PEITC-treated hepatocytes demonstrated that SFN greatly decreased cytochrome P450 (CYP) 3A4 mRNA but did not induce the expression of either glutathione S-transferase (GST) M1 or GSTT1. The protective effects of SFN required pretreatment; cotreatment of hepatocytes with SFN and AFB in the absence of pretreatment had no effect on AFB-DNA adduct formation. When AFB-DNA adduct formation was evaluated by GST genotype, the presence of one or two functional alleles of GSTM1 was associated with a 75% reduction in AFB-DNA adducts, compared with GSTM1 null. In conclusion, these results demonstrate that the inhibition of AFB-DNA adduct formation by SFN is dependent on changes in gene expression rather than direct inhibition of catalytic activity. Transcriptional repression of genes involved in AFB bioactivation (CYP3A4 and CYP1A2), but not transcriptional activation of GSTs, may be responsible for the protective effects of SFN. Although GSTM1 expression was not induced by SFN, the presence of a functional GSTM1 allele can afford substantial protection against AFB-DNA damage in human liver. The downregulation of CYP3A4 by SFN may have important implications for drug interactions. PMID:20442190

  20. Use of a rainbow trout oligonucleotide microarray to determine transcriptional patterns in aflatoxin B1-induced hepatocellular carcinoma compared to adjacent liver.

    Science.gov (United States)

    Tilton, Susan C; Gerwick, Lena G; Hendricks, Jerry D; Rosato, Caprice S; Corley-Smith, Graham; Givan, Scott A; Bailey, George S; Bayne, Christopher J; Williams, David E

    2005-12-01

    Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, and its occurrence is associated with a number of environmental factors including ingestion of the dietary contaminant aflatoxin B(1) (AFB(1)). Research over the last 40 years has revealed rainbow trout (Oncorhynchus mykiss) to be an excellent research model for study of AFB(1)-induced hepatocarcinogenesis; however, little is known about changes at the molecular level in trout tumors. We have developed a rainbow trout oligonucleotide array containing 1672 elements representing over 1400 genes of known or probable relevance to toxicology, comparative immunology, carcinogenesis, endocrinology, and stress physiology. In this study, we applied microarray technology to examine gene expression of AFB(1)-induced HCC in the rainbow trout tumor model. Carcinogenesis was initiated in trout embryos with 50 ppb AFB(1), and after 13 months control livers, tumors, and tumor-adjacent liver tissues were isolated from juvenile fish. Global gene expression was determined in histologically confirmed HCCs compared to noncancerous adjacent tissue and sham-initiated control liver. We observed distinct gene regulation patterns in HCC compared to noncancerous tissue including upregulation of genes important for cell cycle control, transcription, cytoskeletal formation, and the acute phase response and downregulation of genes involved in drug metabolism, lipid metabolism, and retinol metabolism. Interestingly, the expression profiles observed in trout HCC are similar to the transcriptional signatures found in human and rodent HCC, further supporting the validity of the model. Overall, these findings contribute to a better understanding of the mechanism of AFB(1)-induced hepatocarcinogenesis in trout and identify conserved genes important for carcinogenesis in species separated evolutionarily by more than 400 million years.

  1. Protective effect of Ocimum sanctum on 3-methylcholanthrene, 7,12-dimethylbenz(a)anthracene and aflatoxin B1 induced skin tumorigenesis in mice

    International Nuclear Information System (INIS)

    A study on the protective effect of alcoholic extract of the leaves of Ocimum sanctum on 3-mthylcholanthrene (MCA), 7,12-dimethylbenzanthracene (DMBA) and aflatoxin B1 (AFB1) induced skin tumorigenesis in a mouse model has been investigated. The study involved pretreatment of mice with the leaf extract prior to either MCA application or tetradecanoyl phorbol acetate (TPA) treatment in a two-stage tumor protocol viz a viz, DMBA/TPA and AFB1/TPA. The results of the present study indicate that the pretreatment with alcoholic extract of the leaves of O. sanctum decreased the number of tumors in MCA, DMBA/TPA and AFB1/TPA treated mice. The skin tumor induced animals pretreated with alcoholic extract led to a decrease in the expression of cutaneous γ-glutamyl transpeptidase (GGT) and glutathione-S-transferase-P (GST-P) protein. The histopathological examination of skin tumors treated with leaf extract showed increased infiltration of polymorphonuclear, mononuclear and lymphocytic cells, decreased ornithine decarboxylase activity with concomitant enhancement of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the serum, implying the in vivo antiproliferative and immunomodulatory activity of leaf extract. The decrease in cutaneous phase I enzymes and elevation of phase II enzymes in response to topical application of leaf extract prior to MCA, AFB1, DMBA/TPA and AFB1/TPA treatment indicate the possibility of impairment in reactive metabolite(s) formation and thereby reducing skin carcinogenicity. Furthermore, pretreatment of leaf extract in the carcinogen induced animals resulted in elevation of glutathione levels and decrease in lipid peroxidation along with heat shock protein expression, indicating a scavenging or antioxidant potential of the extract during chemical carcinogenesis. Thus it can be concluded that leaf extract of O. sanctum provides protection against chemical carcinogenesis in one or more of the following mechanisms: (i) by acting as an

  2. Effects of astaxanthin and esterified glucomannan on hematological and serum parameters, and liver pathological changes in broilers fed aflatoxin-B1-contaminated feed.

    Science.gov (United States)

    Cao, Jigang; Wang, Wenjun

    2014-02-01

    The effects of astaxanthin (ASTA) and esterified glucomannan (EMG) on hematological and serum parameters, and liver pathological changes in broilers fed on aflatoxin-B1 (AFB1) contaminated diet were investigated. Two hundred and forty 10-day-old broilers were randomly assigned to one of five dietary treatments including: (i) control diet; (ii) AFB1-contaminated diet; (iii) AFB1 + EGM diet; (iv) AFB1 + ASTA diet; and (v) AFB1 + EGM + ASTA diet. At 35 days old, blood and liver tissue samples were collected for analysis. Results indicated that total white blood cell (WBC) number, hemoglobin (Hgb) concentration, hematocrit (Hct) level, serum alanine amino transferase (AST) and γ-glutamyl transferase (GGT) activities, red blood cell (RBC) number, serum globulin (GLB) and urea nitrogen (BUN) concentrations (P contaminated diet. EMG and ASTA alleviated the alteration of RBC, WBC, Hgb and AST caused by AFB1-contaminated diet. Liver superoxide dismutase (SOD) activity was reduced, while myeloperoxidase (MPO) activity was increased by AFB1-contaminated diet (P contaminated diet. It suggested that feeding 0.4 mg/kg AFB1-contaminated diet resulted in adverse effects on blood parameters and liver morphology. Dietary addition of EGM addition at 5 g/kg diet, ASTA at 10 mg/kg diet and especially their combination showed positive protection effects on alleviating the alteration of feeding AFB1. The results indicated that supplementation of 5 g EGM/kg diet, 10 mg ASTA/kg diet and their combination could partially or greatly alleviate the adverse effects caused by AFB1, with the EGM+ASTA group receiving the most effective treatment.

  3. Assessment of Multi-Mycotoxin Exposure in Southern Italy by Urinary Multi-Biomarker Determination

    OpenAIRE

    Solfrizzo, Michele; Gambacorta, Lucia; Visconti, Angelo

    2014-01-01

    Human exposure assessment to deoxynivalenol (DON), aflatoxin B1 (AFB1), fumonisin B1 (FB1), zearalenone (ZEA) and ochratoxin A (OTA) can be performed by measuring their urinary biomarkers. Suitable biomarkers of exposure for these mycotoxins are DON + de-epoxydeoxynivalenol (DOM-1), aflatoxin M1 (AFM1), FB1, ZEA + α-zearalenol (α-ZOL) + β-zearalenol (β-ZOL) and OTA, respectively. An UPLC-MS/MS multi-biomarker method was used to detect and measure incidence and levels of these biomarkers in ur...

  4. Comparison of methods by TLC and HPTLC for determination of aflatoxin M1 in milk and B1 in eggs Comparação de metodologia para análise de aflatoxina M1 em leite e aflatoxina B1 em ovos por CCD e CCDAE

    OpenAIRE

    Scussel, V. M.

    2003-01-01

    Milk and egg matrixes were assayed for aflatoxin M1 (AFM1) and B1 (AFB1) respectively, by AOAC official and modified methods with detection and quantification by thin layer chromatography (TLC) and high performance thin layer chromatography (HPTLC). The modified methods: Blanc followed by Romer, showed to be most appropriate for AFM1 analysis in milk. Both methods reduced emulsion formation, produced cleaner extracts, no streaking spots, precision and accuracy improved, especially when quanti...

  5. 免疫层析试纸法快速检测粮食中黄曲霉毒素B1的验证研究%VERIFICATION OF RAPID DETECTION OF AFLATOXIN B1 IN GRAIN BY IMMUNOCHROMATOGRAPHIC TEST PAPER METHOD

    Institute of Scientific and Technical Information of China (English)

    刘坚; 熊宁; 刘利; 余敦年

    2011-01-01

    4个国家粮食质量监测机构,会同两家免疫层析试纸开发厂商,使用稻谷、玉米两种受黄曲霉毒素B1污染的原始样本,用高效液相色谱法和免疫层析试纸法对粮食中黄曲霉毒素B1测定方法进行了对照研究,对免疫层析试纸法进行了可行性验证.结果表明:免疫层析试纸法和高效液相色谱法的相符率可达90.5%以上;免疫层析试纸法操作简单,检测准确、方便、快速,可用于现场快速检测和初筛粮食中的黄曲霉毒素B1.%In this paper,four state grain and oil inspection mechanisms carried out the feasibility verification of immunochrornatographic test paper method by comparing the effects of high performance liquid chromatography and the immunochromatographic test paper method on determining the aflatoxin B1 in two original samples,I.e. Rice and com, contaminated by aflatoxin B1. The results showed that the consistence rate of the immunochromatographic test paper method and the high performance liquid chromatography was up to 90.5% ;and the immunochromatographic test paper method was simple,accurate,convenient and rapid,and could be used for field rapid determination of aflatoxin B1 in preliminarily screened grain.

  6. 2009-2013年广州市市售粮油食品黄曲霉毒素B1调查%Analysis on contamination of aflatoxin B1 in food and oil in Guangzhou from 2009 to 2013

    Institute of Scientific and Technical Information of China (English)

    张维蔚; 何洁仪; 李迎月; 余超; 林晓华; 梁伯衡

    2015-01-01

    Objective To investigate the contamination of aflatoxin B1 in food and oil in Guangzhou,and provide the basic data of dietary intakes of aflatoxin B1 for food safety assessment.Methods The samples of seven kinds of food including rice,wheat flour,peanut and corn oil,peanut,corn flour,fried food and soybean from ten regions were collected randomly from farmer's markets,supermarkets,wholesale markets,and catering units.The national standard detection method for aflatoxin B1 (ELISA) was taken to detect the content of aflatoxin B1.Results 260 samples were detected aflatoxin B1 and the detection rate was 31.71%,and the farmer's markets had the highest detection rate.The detection range was 0.025 ~ 39.300 μg/kg,with the mean value of 2.675 μg/kg and the median of 2.5 μg/kg.The overall qualified rate was 98.66%.Conclusion The overall level of aflatoxin B1 contamination in market food was low,but some foods such as vegetable oils (peanut,corn) should be more concerned.%目的 了解2009-2013年广州市市售食品中黄曲霉毒素B1污染水平,为广州市开展居民膳食中黄曲霉毒素B1风险评估提供基础数据.方法 在广州市10个区的农贸市场、超市、批发市场、餐饮单位、加工场以及零售店等随机采集米及米制品(大米及米粉)、面及面制品(小麦粉及面包)、植物油(花生油、玉米油)、花生(熟制及生花生)、玉米粉(渣、碎)、油炸食品以及大豆共7类食品,采用国家标准测定方法(ELISA)进行黄曲霉毒素B1的含量测定.结果 820份样品中共260份被检出黄曲霉毒素B1,检出率为31.71%,检出值范围为0.012 ~39.300 μg/kg,均值为2.675 μg/kg,中位数为2.5μg/kg,食品总体合格率为98.66%.结论 广州市市售粮油食品黄曲霉毒素B1总体污染水平不高,但植物油(花生、玉米)检出率较高.

  7. Method Validation for the Quantitative Analysis of Aflatoxins (B1, B2, G1, and G2) and Ochratoxin A in Processed Cereal-Based Foods by HPLC with Fluorescence Detection.

    Science.gov (United States)

    Gazioğlu, Işil; Kolak, Ufuk

    2015-01-01

    Modified AOAC 991.31 and AOAC 2000.03 methods for the simultaneous determination of total aflatoxins (AFs), aflatoxin B1, and ochratoxin A (OTA) in processed cereal-based foods by RP-HPLC coupled with fluorescence detection were validated. A KOBRA® Cell derivatization system was used to analyze total AFs. One of the modifications was the extraction procedure of mycotoxins. Both AFs and OTA were extracted with methanol-water (75+25, v/v) and purified with an immunoaffinity column before HPLC analysis. The modified methods were validated by measuring the specificity, selectivity, linearity, sensitivity, accuracy, repeatability, reproducibility, recovery, LOD, and LOQ parameters. The validated methods were successfully applied for the simultaneous determination of mycotoxins in 81 processed cereal-based foods purchased in Turkey. These rapid, sensitive, simple, and validated methods are suitable for the simultaneous determination of AFs and OTA in the processed cereal-based foods.

  8. Method Validation for the Quantitative Analysis of Aflatoxins (B1, B2, G1, and G2) and Ochratoxin A in Processed Cereal-Based Foods by HPLC with Fluorescence Detection.

    Science.gov (United States)

    Gazioğlu, Işil; Kolak, Ufuk

    2015-01-01

    Modified AOAC 991.31 and AOAC 2000.03 methods for the simultaneous determination of total aflatoxins (AFs), aflatoxin B1, and ochratoxin A (OTA) in processed cereal-based foods by RP-HPLC coupled with fluorescence detection were validated. A KOBRA® Cell derivatization system was used to analyze total AFs. One of the modifications was the extraction procedure of mycotoxins. Both AFs and OTA were extracted with methanol-water (75+25, v/v) and purified with an immunoaffinity column before HPLC analysis. The modified methods were validated by measuring the specificity, selectivity, linearity, sensitivity, accuracy, repeatability, reproducibility, recovery, LOD, and LOQ parameters. The validated methods were successfully applied for the simultaneous determination of mycotoxins in 81 processed cereal-based foods purchased in Turkey. These rapid, sensitive, simple, and validated methods are suitable for the simultaneous determination of AFs and OTA in the processed cereal-based foods. PMID:26268976

  9. 饲喂不同浓度黄曲霉毒素B1饲料对异育银鲫成鱼的生长和毒素积累的影响%GROWTH AND AFLATOXIN B1 ACCUMULATION OF GIBEL CARP ADULT FED WITH DIETS OF DIFFERENT LEVELS OF AFLATOXIN B1

    Institute of Scientific and Technical Information of China (English)

    黄莹; 朱晓鸣; 韩冬; 杨云霞; 金俊琰; 李海燕; 陈毅峰; 解绶启

    2012-01-01

    A 56-day feeding trial was conducted to evaluate the effect of dietary aflatoxin B[ (AFB!) on growth, physiological responses, histological changes, and accumulation in gibel carp (Carassius auratus gibelio) adult. Triplicate groups of gibel carp [(122.3 ?0.7) g] were fed with five semi-purified diets (Diet 1 to 5) containingO, 5, 20, 50 and 500 ug/kg AFB (determined level was 2.59, 4.12, 12.39, 46.23 and 454.07 ug AFB^kg diet, respectively). During the experiment, photoperiod was 12D : 12L with the light period from 09:00 to 21:00, dissolved oxygen was above 6 mg/L, ammonia-N was less than 0.4 mg/L and pH was about 6.8. The results showed that during the 56-day of AFB, exposure, no external changes and unusual behavior were observed in the fish fed with various levels of AFBj. The survival rate in all groups attained 100%. There was no significant difference in final body weight, feeding rate (FR), specific growth rate (SGR) and feed efficiency (FE) between the control and the other groups. Fish fed with various levels of AFB, showed no significant differences in total cholesterol, activities of serum alanine aminotransferase (ALT), asparitic aminotransferase (AST), alkaline phosphatase (ALP) and superoxide dismutase (SOD) compared to the control group. No significant histological lesions in hepatopancreasand ovary were identified between the control and increasing AFB] treatments. Low AFB! residues were found in muscles and ovary, and were below the safety limitation of 5 ^g/kg Accumulation of AFBi in hepatopancreas was logarithmically correlated to the dietary AFB| level. Our results indicated that gibel carp was a less susceptible species to AFBi exposure up to approximately 500 ug/kg diet (determined level was 454.07 ug/kg diet), at least for 56 days.%以含不同浓度黄曲霉毒素B1(AFB1)的配合饲料饲喂异育银鲫(Carassius auratus gibelio)成鱼56d,研究异育银鲫成鱼[(122.3±0.7)g]生长、生理反应、肝脏组织学变化、卵巢发育

  10. Fabrication of a novel nanocomposite based on sol-gel process for hollow fiber-solid phase microextraction of aflatoxins: B1 and B2, in cereals combined with high performane liquid chromatography-diode array detection.

    Science.gov (United States)

    Es'haghi, Zarrin; Sorayaei, Hoda; Samadi, Fateme; Masrournia, Mahboubeh; Bakherad, Zohreh

    2011-10-15

    The new pre-concentration technique, hollow fiber-solid phase microextraction based on carbon nanotube reinforced sol-gel and liquid chromatography-photodiode array detection was applied to determination of aflatoxins B(1), B(2) (AFB(1), AFB(2)) in rice, peanut and wheat samples. This research provides an overview of trends related to synthesis of solid phase microextraction (SPME) sorbnents that improves the assay of aflatoxins as the semi-polar compounds in several real samples. It mainly includes summary and a list of the results for a simple carbon nanotube reinforced sol-gel in-fiber device. This device was used for extraction, pre-concentration and determination of aflatoxins B1, B2 in real samples. In this technique carbon nanotube reinforced sol was prepared by the sol-gel method via the reaction of phenyl trimethoxysilane (PTMS) with a basic catalyst (tris hydroxymethyl aminomethan). The influences of microextraction parameters such as pH, ageing time, carbon nanotube contents, desorption conditions, desorption solvent and agitation speed were investigated. Optimal HPLC conditions were: C(18) reversed phase column for separation, water-acetonitril-methanol (35:10:55) as the mobile phase and maximum wavelength for detection was 370 nm. The method was evaluated statistically and under optimized conditions, the detection limits for the analytes were 0.074 and 0.061 ng/mL for B1 and B2 respectively. Limit of quantification for B1 and B2 was 0.1 ng/mL too (n=7). The precisions were in the range of 2.829-2.976% (n=3), and linear ranges were within 0.1 and 400 ng/mL. The method was successfully applied to the analysis of cereals (peanut, wheat, rice) with the relative recoveries from 47.43% to 106.83%. PMID:21925977

  11. Activity of the aqueous extract from Polymnia sonchifolia leaves on growth and production of aflatoxin B1 by Aspergillus flavus Atividade do extrato aquoso de folhas de Polymnia sonchifolia no crescimento e produção de aflatoxina B1 por Aspergillus flavus

    Directory of Open Access Journals (Sweden)

    Marina M. Pinto

    2001-06-01

    Full Text Available The aqueous extract from Polymnia sonchifolia leaves (AE was tested for inhibitory activity on aflatoxin B1(AFB1 production and growth of Aspergillus flavus. The cytotoxicity of AE on Vero cells was also performed. Suspensions of A. flavus spores were inoculated into 50 mL of YES medium together with different concentrations of the AE. The aflatoxin B1 was extracted, analyzed by thin layer chromatography and quantified by photodensitometry. All the concentrations of AE induced inhibition of AFB1 production. The aqueous extract showed in vitro cytotoxicity to Vero cells only at concentrations above 500 µg/mL.Neste trabalho verificou-se a atividade do extrato aquoso de folhas de Polymnia sonchifolia no crescimento e na produção de aflatoxinas B1 por Aspergillus flavus. Suspensões de esporos de A. flavus foram inoculadas em 50 mL de meio de YES com diferentes concentrações do extrato aquoso. A aflatoxina B1 foi extraída e analisada por cromatografia de camada delgada e quantificada por fotodensitometria. Todas as concentrações testadas inibiram a produção de aflatoxina B1. O extrato aquoso apresentou citotoxicidade em células Vero somente em concentrações acima de 500 µg/mL.

  12. 酶联免疫定量测试盒法测定黄曲霉毒素B_1的方法探讨%Discussion about the Determination of Aflatoxin B1 by ELISA and the Quantitative Test Kit Method

    Institute of Scientific and Technical Information of China (English)

    黄凤妹

    2011-01-01

    This article was studied pH、cations(such as Cu2+,Fe3+,etc.)、coloring solution(such as red yeast) to explore the three ELISA test kit method to measure Aflatoxin B1 samples when the extraction of aflatoxin B1 Extracted with methanol solution method prone t%本文主要通过pH值、阳离子(如Cu2+、Fe3+等)、溶液呈色(如红曲)三方面来探讨酶联免疫测试盒法测定黄曲霉毒素B1时,用甲醇水溶液提取样品中黄曲霉毒素B1的提取方法易出现假阳性,干扰测定,必须进一步用三氯甲烷来提取黄曲霉毒素B1以消除假阳性、排除干扰,提高测定数据的准确性。

  13. 中药材中黄曲霉毒素B1,B2,G1,G2测定方法研究%Determination Method of Aflatoxins B1,B2,G1,G2 in Chinese Medicinal Herbs

    Institute of Scientific and Technical Information of China (English)

    李浩

    2015-01-01

    目的:建立测定中药材中黄曲霉毒素B1,B2,G1,G2的高效液相色谱分离-串联三重四级杆质谱分析(HPLC-MS/MS)法。方法中药材样本经70%甲醇溶液提取,并采用免疫亲和柱净化和浓缩后,以HPLC-MS/MS对4个黄曲霉毒素的含量进行测定。结果黄曲霉毒素G2和B2进样量在0.3~30 pg、黄曲霉毒素G1和B1进样量在1~100 pg范围内与峰面积呈良好线性关系( r﹥0.9990),加样回收率为75.33%~92.29%。结论该法速度快、操作简便、灵敏度高、重复性好,适用于中药材中霉菌毒素的检测。%Objective To establish a HPLC combined with electro spray ionization triple quadrupole tandem mass spectrometry(HPLC-MS/MS)method for determining aflatoxins B1,B2,G1 and G2 in Chinese medicinal herbs. Methods After extraction by 70% methanol and purification and concentration by immunoaffinity column in the Chinese medicinal herb sample,the contents of aflatoxins B1,B2,G1 and G2 were analyzed by HPLC-MS/MS. Results The linear range was 0. 3-30 pg for aflatoxins G2 and B2,1-100 pg for aflatox-ins G1 and B1 ( r ﹥ 0. 999 0 ) . The recovery rate was 75. 33% -92. 29%. Conclusion The method is accurate,simple to operate,highly sensitive,better reproducible and suitable for the aflatoxins determination of Chinese medicinal herbs.

  14. Sampling and analysis of aflatoxin B1 in edible vegetable oil and rice sold in cities of Guangxi region%广西地区市售食用植物油和大米中黄曲霉毒素B1的采样调查和分析

    Institute of Scientific and Technical Information of China (English)

    滕南雁; 宋宁宁; 刘涛

    2011-01-01

    Objective:To investigate the contaminations of aflatoxin B1 in edible vegetable oil and rice sold in supermarkets,markets and fairs in the Guangxi region, and to analyze the potential hazards. Methods: Assay the aflatoxin B1 in terms of the method described in the national standard (ELISA). Results: The aflatoxin B1 present in the edible vegetable oil samples (AFT positive) at 2 μg/kg to 160 μg/kg, with an excess of 35.7% above the limit; while the aflatoxin B1 ( AFT positive) present in the rice samples at 1 μg/kg, meeting the requirement. Conclusion: Edible vegetable oil risks aflatoxin B1 contamination on bases of the data obtained from the tested samples, in contrast with rice.%目的:采集广西区内超市、市场、集市销售的食用植物油和大米,进行黄曲霉毒素B的污染情况调查,了解广西地区市售食用植物油和大米黄曲霉毒素B中污染状况,对其潜在的危害进行分析.方法:采用国家标准测定方法(ELISA)进行黄曲霉毒素B的含量测定.结果:食用植物油中黄曲霉毒素B(AFT阳性)污染量在2 μg/kg至160 μg/kg,超标率为35.7%;检出大米中黄曲霉毒素B(AFT阳性)污染量为1 μg/kg,符合国家规定.结论:样本中食用植物油含黄曲霉毒素B显示存在一定的食品安全风险,大米情况较好.

  15. Determination Method of Aflatoxins B1,B2,G1,G2 in Chinese Medicinal Herbs%中药材中黄曲霉毒素B1,B2,G1,G2测定方法研究

    Institute of Scientific and Technical Information of China (English)

    李浩

    2015-01-01

    Objective To establish a HPLC combined with electro spray ionization triple quadrupole tandem mass spectrometry(HPLC-MS/MS)method for determining aflatoxins B1,B2,G1 and G2 in Chinese medicinal herbs. Methods After extraction by 70% methanol and purification and concentration by immunoaffinity column in the Chinese medicinal herb sample,the contents of aflatoxins B1,B2,G1 and G2 were analyzed by HPLC-MS/MS. Results The linear range was 0. 3-30 pg for aflatoxins G2 and B2,1-100 pg for aflatox-ins G1 and B1 ( r ﹥ 0. 999 0 ) . The recovery rate was 75. 33% -92. 29%. Conclusion The method is accurate,simple to operate,highly sensitive,better reproducible and suitable for the aflatoxins determination of Chinese medicinal herbs.%目的:建立测定中药材中黄曲霉毒素B1,B2,G1,G2的高效液相色谱分离-串联三重四级杆质谱分析(HPLC-MS/MS)法。方法中药材样本经70%甲醇溶液提取,并采用免疫亲和柱净化和浓缩后,以HPLC-MS/MS对4个黄曲霉毒素的含量进行测定。结果黄曲霉毒素G2和B2进样量在0.3~30 pg、黄曲霉毒素G1和B1进样量在1~100 pg范围内与峰面积呈良好线性关系( r﹥0.9990),加样回收率为75.33%~92.29%。结论该法速度快、操作简便、灵敏度高、重复性好,适用于中药材中霉菌毒素的检测。

  16. 应用流式微球检测黄曲霉毒素B1方法的建立%Determination of aflatoxin B1 based on a flow cytometric microsphere immunoassay

    Institute of Scientific and Technical Information of China (English)

    李泳宁; 吴海燕; 郑允权; 郭养浩

    2012-01-01

    采用活性酯法,将AFB1-BSA人工抗原交联于含有羧基表面的荧光微球,通过与游离AFB1竞争抗AFB1单克隆抗体后,再与异硫氰酸荧光素标记二抗的反应,建立基于微球的间接竞争免疫检测方法.检测结果表明,流式细胞仪检测AFB1的检测限为0.03 ng·mL-1,检测范围为0.05 ~ 1.0 ng· mL-1.与黄曲霉毒素B2、黄曲霉毒素G1、黄曲霉毒素G2、黄曲霉毒素M1和黄曲霉毒素M2的交叉反应率均低于1.0%.在玉米样品中的加标回收率为87% ~ 103%,变异系数为6.1%~8.4%.%Carboxyl - modified microspheres internally dyed with fluroscent dye were conjugated with the artificial antigen AFB, - BSA. Aflatoxin B, ( AFB,) was used as a positive control to compete with the AFB, - BSA antigen on the surface of the microspheres for the anti - AFB, McAb. A fluorescein isothiocyanate labeled IgG reporter antibody was added to react specifically with the anti - AFB, McAb on the microspheres. The detection limit of AFB, reached 0.03 ng · mL-1, with a good linearity ranging 0.05 ~ 1.0 ng mL-1. The cross - reactivity rates were less than 1.0% with other toxins such as aflatoxins B2, aflatoxins G,, aflatoxins G2, aflatoxins M, and aflatoxins M2. The recovery of AFB, from artificially contaminated corn samples was from 89% to 92% , with CVs from 6.8% to 9.0%. A novel method for the determination of aflatoxin B, by an indirect competitive immunoassay with a flow cytometer has been developed.

  17. Determination of aflatoxins B1, B2, G1, G2 in food by UPLC%UPLC同时测定食品中黄曲霉毒素B1、B2、G1、G2

    Institute of Scientific and Technical Information of China (English)

    丘汾; 李可; 周海涛; 刘奋; 杨梅; 曾胜波; 戴京晶

    2012-01-01

    Objective; A simple and rapid method for determination of aflatoxin B, ,B2 ,G, and G2 content in food by UPLC were developed. Methods; After extraction with MeOH - H2O,the extracts of food homogenization was loaded on the I AC for purification and determined by UPLC. Results; The recoveries of samples were in the range of 75.0% -90.4% ,the RSD was below 5% . The limit of quantification for aflatoxin B, was 0. 2 μg/kg,the limit of quantification for aflatoxin B2 was 0.2 μg/kg, the limit of quantification for aflatoxin G, was 0.4 μg/kg, the limit of quantification for aflatoxin G2 was 0.2 μg/kg. Conclusion; Determination of aflatoxins B, ,B2 ,G,and G2 content in food by immune affinity column was a rapid,simple and exact method.%目的:建立免疫亲和柱超高效液相色谱法测定食品中黄曲霉毒素B1,B2,G1和G2含量的方法.方法:试样经过甲醇-水提取、稀释后经过免疫亲和柱层析净化,应用超高液相色谱法检测.结果:试验结果表明:空白样品分别按照0.2 μg/kg、0.8μg/kg、2.0μg/kg添加黄曲霉毒素混合标准,回收率为75.0%~90.4%,精密度<5%,黄曲霉毒素B1,B2,G1和G2的检测灵敏度分别为0.2μg/kg,0.2 μg/kg,0.4 μg/kg,0.2 μg/kg.结论:免疫亲和柱净化超高效液相色谱法测定食品中黄曲霉毒素B1,B2,G1,G2含量,是一种简单、快速和准确的方法.

  18. Metabolomics of the Bio-Degradation Process of Aflatoxin B1 by Actinomycetes at an Initial pH of 6.0

    Directory of Open Access Journals (Sweden)

    Manal Eshelli

    2015-02-01

    Full Text Available Contamination of food and feed by Aflatoxin B1 (AFB1 is a cause of serious economic and health problems. Different processes have been used to degrade AFB1. In this study, biological degradation of AFB1 was carried out using three Actinomycete species, Rhodococcus erythropolis ATCC 4277, Streptomyces lividans TK 24, and S. aureofaciens ATCC 10762, in liquid cultures. Biodegradation of AFB1 was optimised under a range of temperatures from 25 to 40 °C and pH values of 4.0 to 8.0. An initial concentration of 20 µg/mL of AFB1 was used in this study. The amount of AFB1 remaining was measured against time by thin layer chromatography (TLC and high-performance liquid chromatography (HPLC, coupled with UV and mass spectrometry (LC-MS. All species were able to degrade the AFB1, and no significant difference was found between them. AFB1 remained in the liquid culture for R. erythropolis, S. lividans and S. aureofaciens were 0.81 µg/mL, 2.41 µg/mL and 2.78 µg/mL respectively, at the end of the first 24 h. Degradation occurred at all incubation temperatures and the pH with the optimal conditions for R. erythropolis was achieved at 30 °C and pH 6, whereas for S. lividans and S. aureofaciens the optimum conditions for degradation were 30 °C and pH 5. Analysis of the degradative route indicated that each microorganism has a different way of degrading AFB1. The metabolites produced by R. erythropolis were significantly different from the other two microorganisms. Products of degradation were identified through metabolomic studies by utilizing high-resolution mass spectral data. Mass spectrometric analysis indicated that the degradation of AFB1 was associated with the appearance of a range of lower molecular weight compounds. The pathway of degradation or chemical alteration of AFB1 was followed by means of high resolution Fourier transform mass spectrometry (HR-FTMS analysis as well as through the MS2 fragmentation to unravel the degradative pathway for

  19. Alpha-class glutathione S-transferases in wild turkeys (Meleagris gallopavo: characterization and role in resistance to the carcinogenic mycotoxin aflatoxin B1.

    Directory of Open Access Journals (Sweden)

    Ji Eun Kim

    Full Text Available Domestic turkeys (Meleagris gallopavo are one of the most susceptible animals known to the toxic effects of the mycotoxin aflatoxin B1 (AFB1, a potent human hepatocarcinogen, and universal maize contaminant. We have demonstrated that such susceptibility is associated with the inability of hepatic glutathione S-transferases (GSTs to detoxify the reactive electrophilic metabolite exo-AFB1-8,9-epoxide (AFBO. Unlike their domestic counterparts, wild turkeys, which are relatively AFB1-resistant, possess hepatic GST-mediated AFBO conjugating activity. Here, we characterized the molecular and functional properties of hepatic alpha-class GSTs (GSTAs from wild and domestic turkeys to shed light on the differences in resistance between these closely related strains. Six alpha-class GST genes (GSTA amplified from wild turkeys (Eastern and Rio Grande subspecies, heritage breed turkeys (Royal Palm and modern domestic (Nicholas strain turkeys were sequenced, and catalytic activities of heterologously-expressed recombinant enzymes determined. Alpha-class identity was affirmed by conserved GST domains and four signature motifs. All GSTAs contained single nucleotide polymorphisms (SNPs in their coding regions: GSTA1.1 (5 SNPs, GSTA1.2 (7, GSTA1.3 (3, GSTA2 (3, GSTA3 (1 and GSTA4 (2. E. coli-expressed GSTAs possessed varying activities toward GST substrates 1-chloro-2,4-dinitrobenzene (CDNB, 1,2-dichloro-4-nitrobenzene (DCNB, ethacrynic acid (ECA, cumene hydroperoxide (CHP. As predicted by their relative resistance, livers from domestic turkeys lacked detectable GST-mediated AFBO detoxification activity, whereas those from wild and heritage birds possessed this critical activity, suggesting that intensive breeding and selection resulted in loss of AFB1-protective alleles during domestication. Our observation that recombinant tGSTAs detoxify AFBO, whereas their hepatic forms do not, implies that the hepatic forms of these enzymes are down-regulated, silenced, or

  20. Alpha-class glutathione S-transferases in wild turkeys (Meleagris gallopavo): characterization and role in resistance to the carcinogenic mycotoxin aflatoxin B1.

    Science.gov (United States)

    Kim, Ji Eun; Bunderson, Brett R; Croasdell, Amanda; Reed, Kent M; Coulombe, Roger A

    2013-01-01

    Domestic turkeys (Meleagris gallopavo) are one of the most susceptible animals known to the toxic effects of the mycotoxin aflatoxin B1 (AFB1), a potent human hepatocarcinogen, and universal maize contaminant. We have demonstrated that such susceptibility is associated with the inability of hepatic glutathione S-transferases (GSTs) to detoxify the reactive electrophilic metabolite exo-AFB1-8,9-epoxide (AFBO). Unlike their domestic counterparts, wild turkeys, which are relatively AFB1-resistant, possess hepatic GST-mediated AFBO conjugating activity. Here, we characterized the molecular and functional properties of hepatic alpha-class GSTs (GSTAs) from wild and domestic turkeys to shed light on the differences in resistance between these closely related strains. Six alpha-class GST genes (GSTA) amplified from wild turkeys (Eastern and Rio Grande subspecies), heritage breed turkeys (Royal Palm) and modern domestic (Nicholas strain) turkeys were sequenced, and catalytic activities of heterologously-expressed recombinant enzymes determined. Alpha-class identity was affirmed by conserved GST domains and four signature motifs. All GSTAs contained single nucleotide polymorphisms (SNPs) in their coding regions: GSTA1.1 (5 SNPs), GSTA1.2 (7), GSTA1.3 (3), GSTA2 (3), GSTA3 (1) and GSTA4 (2). E. coli-expressed GSTAs possessed varying activities toward GST substrates 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), ethacrynic acid (ECA), cumene hydroperoxide (CHP). As predicted by their relative resistance, livers from domestic turkeys lacked detectable GST-mediated AFBO detoxification activity, whereas those from wild and heritage birds possessed this critical activity, suggesting that intensive breeding and selection resulted in loss of AFB1-protective alleles during domestication. Our observation that recombinant tGSTAs detoxify AFBO, whereas their hepatic forms do not, implies that the hepatic forms of these enzymes are down-regulated, silenced, or

  1. Effect of melatonin on the production of microsomal hydrogen peroxide and cytochrome P-450 content in rat treated with aflatoxin B(1).

    Science.gov (United States)

    Awney, Hala A; Attih, Ahmed M; Habib, Sami L; Mostafa, Mostafa H

    2002-03-20

    Aflatoxin B(1) (AFB(1)) is a food contaminant fungal toxin that has been implicated as a causative agent in human hepatic and extrahepatic carcinogenesis. In this study we went on to show the effect of melatonin as a free radical scavenger on the production of microsomal hydrogen peroxide (H(2)O(2)) during the metabolic activation AFB(1). The production of microsomal H(2)O(2) in vitro during the metabolic activation of different chemical carcinogens has been reported previously. We also studied the effect of melatonin on the cytochrome P-450 content as a major microsomal monooxygenase isoenzymes system in rat liver responsible for the metabolic activation of AFB(1). The amounts of H(2)O(2) and cytochrome P-450 contents in rat treated with melatonin (0.2 mg/kg BW) and/or AFB(1) (0.2 mg/kg BW) at various time intervals has been measured. Animals treated with melatonin exhibited markedly inhibition in the amounts of H(2)O(2) after 1, 3, and 6 h. The highest level of inhibition (3.0 nmol H(2)O(2)/mg protein) was detected after 6 h. However, cytochrome P-450 contents were also decreased after the same period of time. The highest level of inhibition (2.1 nmol/mg protein) was detected after 3 h of injection. A pronounced augmentation of H(2)O(2) production was observed in rat treated with AFB(1) only. The highest level of H(2)O(2) (100 nmol/mg protein) was measured after 1 h. Cytochrome P-450 contents were also decreased in response to AFB(1) injection over the same time intervals. Contrary data was detected in animals received both AFB(1) and melatonin. The generation of H(2)O(2) was inhibited by melatonin after 1, 3 and 6 h. The highest level of inhibition (44.2 nmol/mg protein) was observed after 6 h. Finally, these data suggested that melatonin as a free radical scavenger inhibited the microsomal production of H(2)O(2) in rat treated with AFB(1).

  2. 免疫亲和柱净化-高效液相色谱-三重串联四级杆质谱法测定食用植物油中的黄曲霉毒素B1%Determination of aflatoxin B1 in edible vegetable oil by immunoaffinity column purification and HPLC-MS/MS

    Institute of Scientific and Technical Information of China (English)

    史俊文; 刘凤芝

    2014-01-01

    Immunoaffinity column purification and HPLC-MS/MS method was established to determine aflatoxin B1 in edible vegetable oil. Aflatoxin B1 was extracted from edible vegetable oil by 70% of meth-anol,then the extract was filtered and purified by immunoaffinity column before being injected into HPLC-MS/MS to determine aflatoxin B1 . The results showed that calibration curve presented good line-ar relationship when the mass concentration of aflatoxin B1 ranged from 0. 2 ng/mL to 10 ng/mL;the limit of detection(S/N=3) and the limit of quantification(S/N=10) of the method were 0. 05 μg/kg and 0. 2 μg/kg respectively,the average recovery of standard addition was in the range of 83. 0% -94. 8%, and the relative standard deviation was less than 7 . 8%.%建立了免疫亲和柱净化-高效液相色谱-三重串联四级杆质谱法测定食用植物油中黄曲霉毒素B1的方法。采用70%甲醇水溶液提取食用植物油中黄曲霉毒素B1,提取液经过滤、免疫亲和柱净化后,上高效液相色谱-三重串联四级杆质谱仪进行测定。结果表明,黄曲霉毒素B1在0.2~10 ng/mL范围内方程线性关系良好,方法检出限(S/N=3)为0.05滋g/kg,定量限(S/N=10)为0.2滋g/kg,平均加标回收率在83.0%~94.8%之间,相对标准偏差小于7.8%。

  3. 广西南宁市健康人群黄曲酶毒素B1暴露影响因素的研究%A research of influencing factors on the aflatoxin B1 exposure to the healthy people lived in the city of Nanning in Guangxi Province

    Institute of Scientific and Technical Information of China (English)

    莫新少; 彭涛; 黎乐群; 陈德凤; 游雪梅

    2009-01-01

    Objective To investigate the influencing factore of aflatoxin B1 (AFB1) exposure in healthy people who lived in aflatoxin high-exposure area and to provide a basis for preventing AFB1 exposure. Methods To the residents lived in the city of Nanning in Cuangxi Province, a questionnaire survey was used to acquaint relevant information. The AFB1-albumin adducts (AAA) levels were screened by enzyme-linked immunosorbent assay and the liver function detected using a Bekman LX20 Chemistry Analyzer and diagnostic agents from Randox, UK. Results AAA level was significantly higher (P<0.05) in the people with lowincome, having habits of smoking, drinking, eating bulk rice, and eating rice that had been saved for a time of more than 30 days, and keeping regularly dining out. Conclusions The level of income and the lifestyle are related with Aflatoxin Bl exposure. The risk factors about Aflatoxin Bl exposure include consumption of bulk rice, regularly dining out, and drinking.%目的 探讨黄曲酶毒素高暴露地区健康人群黄曲酶毒素B1(aflatoxin B1,AFB1)暴露的影响因素,为预防AFB1暴露提供依据.方法 以广西南宁市居民为对象,通过问卷调查了解相关资料,采用竞争酶联免疫法检测血浆黄曲酶毒素B1-白蛋白加合物(AFBI-albumin adducts,AAA)水平,采用Bekman LX20 Chemistry Analyzer配套英国Randox公司诊断试剂检测肝功能.结果 经济收入低、吸烟、饮酒、食用散装大米、大米保存超过30 d、经常在外就餐的人群,AAA水平显著增高(P<0.05).结论 人们经济收入水平及生活方式与AFB1暴露有关,食用散装大米、经常在外就餐、饮酒是AFB1暴露的危险因素.

  4. 高效液相色谱-串联质谱法测定水产品中黄曲霉毒素G2、G1、B2、B1%Determination of Aflatoxin G2、G1、B2、B1 in Aquatic Products with HPLC-MS/MS

    Institute of Scientific and Technical Information of China (English)

    莫彩娜; 杨曦; 黄智成

    2011-01-01

    The method for determining aflatoxin G2,G1,B2,and B1 by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS) was developed.The samples were extracted by 84% methanol,degreased with hexane and purified with HLB solid-phase extraction column.In addition,with electrospray ionization in positive mode,the samples was monitored with multiple reaction monitoring(MRM) and quantified them with external standard method.For the 4 aflatoxins,it showed good linear regression coeffcient in the standard curve(all0.99),the limits of detection for aflatoxin G1,B1 were 0.5 μg/kg,and the limits of detection for aflatoxin G2,B2 were 0.3 μg/kg.The average recovery was 56%~80%,and the relative standard deviation was 1.58%~14.4%.The method,was sensitive and reliable,and can be applied for the determination of aflatoxins in aquatic products.%建立了用高效液相色谱-串联质谱法(HPLC-MS/MS)测定水产品中黄曲霉毒素G2、G1、B2、B1残留量的方法。84%甲醇水溶液提取水产品中4种黄曲霉毒素,正己烷脱脂,HLB固相萃取柱净化。采用电喷雾电离,正离子扫描,选择多反应检测模式(MRM)监测,外标法定量。该法对4种黄曲霉毒素标准曲线的线性回归系数均在0.99以上,黄曲霉毒素G1、B1方法定量限为0.5μg/kg,G2、B2方法定量限为0.3μg/kg。4种黄曲霉毒素的回收率为56%~80%,相对标准偏差1.58%~14.4%。该法灵敏,结果可靠,可用于水产品中黄曲霉毒素的测定。

  5. Simultaneous Determination of Aflatoxins B1, B2, G1 and G2 in Tobacco by HPLC-FLD Pre-column Derivatization Method%HPLC-FLD柱前衍生法同时测定烟草中黄曲霉毒素B1、B2、G1和G2

    Institute of Scientific and Technical Information of China (English)

    王康; 李韵; 肖少红

    2015-01-01

    通过溶剂萃取、免疫亲和柱纯化富集、三氟乙酸柱前衍生、高效液相色谱(HPLC)法分离及荧光检测器检测,建立了同时测定烟草及烟草制品中黄曲霉毒素B1、B2、G1和G2的免疫亲和检测方法.结果表明:①该方法可在20 min内完成测定,4种目标物能够得到很好的分离,线性关系良好,相关系数r值均大于0.99.②方法的回收率为85%~117%,相对标准偏差为0.2%~9.4%(n=6),其中B1的检出限和定量限分别为0.10和0.34μg/kg.%An immunoaffinity detection method for simultaneously determining the aflatoxins B1, B2, G1 and G2 in tobacco and tobacco products was developed via solvent extraction of sample, purifying and concentrating on immunoaffinity column, pre-column derivatization by trifluoroacetic acid, separation by HPLC, and detection by fluorescence detector. The results showed that: 1) The determination could be completed within 20 minutes, the four target aflatoxins were well separated and exhibited good linear relations with correlation coefficients r > 0.99. 2) The recoveries of the method ranged from 85% to 117%with the relative standard deviation (RSD) of 0.2%-9.4% (n=6), and the limits of detection and quantification of aflatoxin B1 were 0.10 and 0.34 μg/kg, respectively.

  6. A contribution to the determination of aflatoxin B1, quinine hydrochloride and L(+)-ascorbic acid in foodstuffs by quantitative in situ thin-layer chromatographic analysis

    NARCIS (Netherlands)

    Beljaars, P.R.

    1974-01-01

    The application of quantitative thin-layer chromatography (TLC) involving in situ spectrodensitometric measurements with a flying-spot densitometer is described in this study for the analysis of aflatoxin B 1 , quinine hydrochloride and L(+)-ascorbic acid (vitamin C

  7. Inactivación de aflatoxina B1 y aflatoxicol por nixtamalización tradicional del maíz y su regeneración por acidificación de la masa Inactivation of aflatoxin B1 and aflatoxicol through traditional "nixtamalización" of corn and their regeneration by acidification of corn dough

    Directory of Open Access Journals (Sweden)

    Gloria Laura Anguiano-Ruvalcaba

    2005-10-01

    Full Text Available OBJETIVO: Confirmar el efecto de la nixtamalización tradicional sobre la aflatoxina, identificar el compuesto remanente en masa, evaluar su toxicidad y su regeneración por tratamiento ácido. MATERIAL Y MÉTODOS: Se utilizó maíz, sin y con aflatoxina, y se nixtamalizó. La toxicidad se evaluó en pollos de ocho días de edad. Se aplicó el tratamiento ácido a la masa. La cuantificación de aflatoxinas se realizó por cromatografía líquida de alta resolución (HPLC. RESULTADOS: La nixtamalización destruyó la aflatoxina (96% y el aflatoxicol (70%; el remanente en masa fue aflatoxina B1. El tratamiento ácido in vitro no eleva las concentraciones de ninguna de las dos micotoxinas. Los pollos murieron al ingerir 260 mg de AFB1, y la masa con aflatoxina remanente no fue tóxica. CONCLUSIONES: Los resultados ilustran el beneficio de la nixtamalización tradicional en la inactivación de las aflatoxinas presentes en maíz y en su no reconstitución por efecto del tratamiento ácido.OBJECTIVE: To ratify the effect of traditional "nixtamalización" upon aflatoxin in corn, to identify the residual compound in dough, to evaluate its toxicity, and to verify the effect of acidic treatment on the regeneration of aflatoxin. MATERIAL AND METHODS: Corn with and without aflatoxin was treated separately with lime and heat to obtain two types of dough. Aflatoxin and the residual compound were quantified by high pressure liquid chromatography (HPLC and toxicity was evaluated in eight day old chicks. RESULTS: "Nixtamalización" destroyed AFB1 (96% and aflatoxicol (70%. The residual compound was identified as AFB1. Experimental animals died by ingestion of 260 mg of AFB1 Dough containing residual aflatoxin was not toxic. Acidic. treatment did not increase aflatoxin concentration, nor aflatoxicol. CONCLUSIONS: The results support the use of traditional "nixtamalización" as a means to inactivate aflatoxin in corn and that acidic treatment prevents its

  8. 高效液相色谱-串联三重四极杆质谱分析法测定刀豆中黄曲霉毒素G2、G1、B2、B1%HPLC-triple quadrupole MS determination of aflatoxin G2,G1,B2,B1 in Canavalia gladiata(Jacq.)

    Institute of Scientific and Technical Information of China (English)

    许勇; 王少敏; 郑荣; 毛丹; 王柯; 季申

    2011-01-01

    目的:建立刀豆中黄曲霉毒素G、G、B、B的测定方法.方法:样品经70%甲醇提取、HLB柱净化后,用高效液相色谱-串联质谱进行分析测定.结果:黄曲霉毒素G、B在0.75~30 pg范围内与峰面积呈良好的线性关系,黄曲霉毒素G、B在2.5 pg~100 pg范围内与峰面积呈良好的线性关系,r>0.999.回收率在66.9%~86.7%之间.结论:本法简便快速、结果准确、重现性好,可用于刀豆中黄曲霉毒素的测定.%Objective:To establish a HPLC - triple quadrupole MS method for determination of aflatoxin G2, G1, B2, B1 in Canavalia gladiata( Jacq.).Methods:After being extracted by 70% methanol, purified by HLB column, aflatoxins were analysed by HPLC- triple quadrupole MS.Results:There is a good linear relationship within the range of 0.75 pg ~30 pg for aflatoxin G2 ,B2, and the range of 2.5 pg ~ 100 pg for aflatoxin G1 ,B1.The recovery was between 66.9% ~86.7%.Conclusion:The method is simple, sensitive, accurate, specific and reproducible in the quality Green Beans.

  9. 超顺磁性免疫磁珠体系用于植物油中黄曲霉毒素B1的检测研究%Research of the Super Paramagnetic Immune Magnetic Beads System for Detection of Aflatoxin B1 in Vegetable Oil

    Institute of Scientific and Technical Information of China (English)

    刘伟伟; 孙秀兰; 张银志; 樊惠良; 陈文君; 李在均

    2011-01-01

    利用化学共沉淀方法,制得了主要成分为Fe3O4的超顺磁性纳米磁珠.纳米磁珠和硅烷偶联剂氨丙基三乙氧基硅烷(APTES)反应而带上氨基基团.带上氨基的磁珠通过戊二醛活化后发生醛基化,与黄曲霉毒素B1( AFB1)多克隆抗体偶联得到黄曲霉毒素B1免疫磁珠.利用该免疫磁珠为净化工具,建立了有机溶剂萃取、免疫磁珠净化、高效液相色谱( HPLC)检测植物油中黄曲霉毒素B1的方法.该方法具有良好线性,线性范围为5~50 μg/L,相关系数达0.999 4,检出限为0.5μg/L,平均回收率为96%,相对标准偏差为12.5%.该方法操作简单、灵敏度高、准确性好,可用于植物油中黄曲霉毒素B1的检测.%Using the method of chemical co-precipitation, the super paramagnetic nanoparticles with the key component of Fe3O4 were synthesized. The magnetic beads reacted with the silane coupling a-gent 3-aminopropyl triethoxysilane and connected with amino groups. After activated by the agent gl-utaraldehyde, the magnetic beads took along the aldehyde group and coupled with aflatoxin B, poly-clonal antibody to form the immune magnetic beads. A method was established to detect the aflatoxin B, in vegetable oil by using the organic solvent for preliminary extraction, the aflatoxin B, immune magnetic beads system to purification and HPLC to quantification. The method showed a good linearity in the range of 5 - 50 p:g/L with correlation coefficient of 0. 999 4 and detection limit of 0. 5 |xg/L. The average recovery was up to 96% and the RSD was 12. 5% . With the advantages of simplicity, sensitivity and high accuracy, the method could be applied in the determination of aflatoxin B1 in vegetable oil.

  10. 高效液相色谱-串联质谱法测定山茶油中黄曲霉毒素B1%Determination of aflatoxin B1in camellia seed oil by liquid chromatography-tandem mass spectrometry

    Institute of Scientific and Technical Information of China (English)

    丁明; 钟冬莲

    2012-01-01

    建立了山茶油中黄曲霉毒素B1含量的高效液相色谱-串联质谱分析方法.通过对前处理方法的优化,选择了甲醇和水作为山茶油中黄曲霉毒素B1的提取溶剂,经免疫亲和柱富集浓缩后,采用高效液相色谱-串联质谱进行分析,经C18色谱柱分离,在电喷雾离子化正离子模式(ESI+)及多反应监测模式(MRM)下进行测定,基质匹配标准溶液外标法定量.在优化条件下,该方法线性范围为0.4 ~6.4 μg/L,相关系数r2>0.998,最低检出限为0.026 μg/kg,在添加水平为0.008,0.016和0.032 μg时,方法回收率在85.9%~93.8%之间;相对标准偏差为1.8% ~5.0%.方法可满足山茶油中黄曲霉毒素B1的检测要求.%In order to improve detection sensitivity and accuracy of aflatoxions, a liquid chromatography-tandem mass spectrometric method was established for qualitative and quantitative analysis of aflatoxins Bl. Samples were simply extracted by methanol-water at a ratio of 70;30 ( V/V). After being filtered,it was cleaned up with an immunoaffinity column. The analysis was realized by multiple reaction-monitoring mode. Under above conditions, aflatoxins Bl could be completely separated within 10 min with an excellent linear relationship. The determination limit for aflatoxin Bl was 0. 012 μg/kg. The recoveries of samples were in the range of 85. 9 % ~ 93. 8 % with a relative standard deviation less than 5%. This method can provide a rapid, sensitive, accurate and reproducible detection for aflatoxin Bl and its detection limit is sensitive enough to meet the European regulation for aflatoxins.

  11. Proteome Profiling of Urinary Exosomes Identifies Alpha 1-Antitrypsin and H2B1K as Diagnostic and Prognostic Biomarkers for Urothelial Carcinoma

    Science.gov (United States)

    Lin, Shih-Yi; Chang, Chao-Hsiang; Wu, His-Chin; Lin, Ching-Chan; Chang, Kai-Po; Yang, Chi-Rei; Huang, Chi-Ping; Hsu, Wu-Huei; Chang, Chiz-Tzung; Chen, Chao-Jung

    2016-01-01

    MALDI-TOF spectrometry has not been used for urinary exosome analysis. We used it for determining UC biomarkers. From 2012 to 2015, we enrolled 129 consecutive patients with UC and 62 participants without UC. Exosomes from their urine were isolated, and analyzed through MALDI-TOF spectrometry. Immunohistochemical (IHC) analysis of another 122 UC and 26 non-UC tissues was conducted to verify the discovered biomarkers. Two peaks at m/z 5593 (fragmented peptide of alpha-1-antitrypsin; sensitivity, 50.4%; specificity, 96.9%) and m/z 5947 (fragmented peptide of histone H2B1K sensitivity, 62.0%; specificity, 92.3%) were identified as UC diagnosis exosome biomarkers. UC patients with detectable histone H2B1K showed 2.29- and 3.11-fold increased risks of recurrence and progression, respectively, compared with those with nondetectable histone H2B1K. Verification results of IHC staining revealed significantly higher expression of alpha 1-antitrypsin (p = 0.038) and H2B1K (p = 0.005) in UC tissues than in normal tissues. The expression of alpha 1-antitrypsin and H2B1K in UC tissues was significantly correlated with UC grades (p exosome proteins alpha 1-antitrypsin and histone H2B1K, which are identified through MALDI-TOF analysis, could facilitate rapid diagnosis and prognosis of UC. PMID:27686150

  12. Determination of Aflatoxin B1, M1 in Foods Using Homemade Mixed Solid Phase Extraction Column Coupled with High Performance Liquid Chromatography%自制混合型固相萃取柱-高效液相色谱法同时测定食品中黄曲霉毒素B1、M1

    Institute of Scientific and Technical Information of China (English)

    彭晓俊; 曾勚; 庞晋山; 邓爱华; 谌瑜

    2013-01-01

    A novel method for the determination of aflatoxin B1,M1 in foods was developed using homemade mixed solid phase extraction column coupled with high performance liquid chromatography.The sample was extracted with 60% acetonitrile solution,and cleaned up on a homemade mixed solid phase extraction column.The separation was performed on a Shim-pack VP-ODS C18 column with acetonitrile-water(20 ∶ 80) as mobile phase within 13 min.The contents of two aflatoxins were detected with fluorescence detector,and quantified by the external standard method.The effects of column type,column capacity,sample amount,extraction solution and flow rate on the determination of aflatoxin residue were investigated.Under the optimized conditions,there were good linearities between peak areas and aflatoxins concentrations in the range of 0.40-100 μg/L with correlation coefficients of 0.999 4-0.999 7.The limits of detection(S/N =3) of aflatoxin B1,M1 were both 0.050 μg/kg.The recoveries of aflatoxins in different samples at three spiked levels of 0.40,1.0,100 μg/L were in the range of 53%-112% with relative standard deviations(RSDs) of 2.7%-7.1%.The proposed method showed the advantages of sensitivity,simplicity and fastness,and was successfully applied in the determination of low aflatoxins residues in peanut,pistachio and milk samples.%建立了基于自制混合型固相萃取柱的样品净化/高效液相色谱测定食品中黄曲霉毒素B1、M1的方法.样品经60%乙腈水溶液提取、离心后,通过自制固相萃取柱排除杂质干扰,流出液以Shim-pack VP-ODS C18色谱柱为分离柱,水和乙腈为流动相,用荧光检测器检测,外标法定量.考察了柱类型、柱容量、取样量、提取溶液和流速等对检测的影响,优化了实验条件.在优化条件下,2种毒素在0.40~100 μg/L质量浓度范围内与峰面积呈良好的线性关系,相关系数为0.9994~0.9997,检出限(S/N=3)为0.050 μg/kg.在样品中分别加入0.40、1

  13. HPLC-MS/MS determination of aflatoxinG2,G1,B2,B1 in Persicae Semen%HPLC-MS/MS法测定中药桃仁中黄曲霉毒素G2、G1、B2、B1

    Institute of Scientific and Technical Information of China (English)

    王少敏; 许勇; 毛丹; 郑荣; 王柯; 季申

    2011-01-01

    目的:建立中药桃仁中黄曲霉毒素G2、G1、B2、Bl的HPLC-MS/MS测定方法.方法:样品经有机溶剂提取及免疫亲和柱净化后,以高效液相色谱-串联三重四极杆质谱进行分析测定.采用Phenomenex SB -C18(2.0mmx50mm,4μm)色谱柱,流动相为甲醇-10mmol·L-1醋酸铵溶液,梯度洗脱,流速0.5mL·min-1;质谱条件为电喷雾离子源(ESI源),正离子模式检测,扫描方式为多反应监测(MRM),黄曲霉毒素G2、G1、B2、B1的定量分析离子分别为m/z 331.1→313.l、m/z 329.1→243.1、m/z 315.0→287.1、m/z 313.1→241.0.结果:黄曲霉毒素G2、B2进样量在0.375-30pg范围内与峰面积呈良好的线性关系,黄曲霉毒素G1、1进样量在1.2-1100 pg范围内与峰面积呈良好的线性关系,0 999;回收率在88.%-1103.%%之间.结论:本法灵敏、快速、准确,专属性强,可用于中药桃仁中黄曲霉毒素的测定.%Objective : To establish an HPLC - MS/MS method for determination of aflatoxin G2 ,G1 , B2 ,B1 in Persicae Semen. Methods :Aflatoxins were extracted by 70% methanol, purified by an immunoaffinity column and then separation was carried on a Kromasil C18 (2. 0 mm ×50 mm ,4 μm) column with a mobile phase of methanol - 10 mmol · L-1 formic ammonate ( by a gradient program) at a rate of 0. 5 mL · min-1. Aflatoxins were analysed by HPLC - triple quadrupole MS with an ESI ion source in MRM mode, the ion combinations of m/z 331. 1→ 313. 1 ,m/z 329. 1→243. 1 , m/z 315. 0→287. 1 , m/z 313. 1→241. 0 were used to qualify aflatoxin G2 , G1 , B2 , B1 , respectively. Results : Good linear relationships were obtained within the ranges of 0. 375 - 30 pg for aflatoxin G2 and B2 ,and 1. 25 - 100 pg for aflatoxin G1 and B1. The recoveries were between 88. 5 % - 103. 9% . Conclusion : The method is sensitive, rapid , accurate and specific for the determination of aflatoxins in traditional Chinese medicine Persicae Semen.

  14. 高效液相色谱柱后光化学反应-荧光检测茶叶中黄曲霉毒素 B1%Determination of Aflatoxins B1 in Tea by High Performance Liquid Chromatography-Fluorescence Detector with Post-column Photochemical Reaction

    Institute of Scientific and Technical Information of China (English)

    赵浩军; 王坤; 杨卫花; 杨朝义

    2013-01-01

      建立了高效液相色谱/光化学反应器/荧光检测器测定茶叶中黄曲霉毒素 B1的方法。用乙腈水溶液(V∶V=86∶14)提取黄曲霉毒素 B1,提取液经净化柱和黄曲霉毒素 B1免疫亲和柱净化,高效液相色谱测定。在黄曲霉毒素 B1标准溶液质量浓度为0.591~5.91μg/L 时,峰面积与浓度呈现良好的线性关系,黄曲霉毒素 B1的回收率为85.4%~98.9%(添加量分别为0.510μg/kg、7.090μg/kg 和14.180μg/kg),相对标准偏差为0.2%~1.8%,方法检出限为0.1μg/kg。运用所建立方法对市售的8个茶样及加标样品中的黄曲霉毒素 B1进行检测,结果显示该方法选择性强、灵敏度高,适合茶叶中黄曲霉毒素 B1的测定。%A new method for the sensitive determination of Aflatoxins B1 in tea by high performance liquid chromatography with photoelectric reactor and fluorescence detector was established. A solution of V(acetonitrile)∶V(H2O)=86∶14 was used to extract Aflatoxins B1 from tea. The extracted solution was then purified by a multifunctional and immuneaffinity column, respectively. The peak area and the concentration of Aflatoxins B1 showed a good linear relationship within the range from 0.591 μg/L to 5.91 μg/L with a linear correlation coefficient (r) of 0.9994. The recoveries at the concentrations studied [low level (7.090 μg/kg), high level (14.180 μg/kg)] were between 85.6% and 98.9% with a relative standard deviations ranging from 1.7% to 1.8%. The limit of detection (LOD) is 0.1 μg/kg (S/N=3). The limit of quantitation (LOQ) is (0.591 μg/kg). The new method was used to analyze eight tea samples collected from the local markets and negative results were obtained. The method is suitable for detection of Aflatoxins B1 in tea with high selectivity and sensitivity.

  15. Effect of gamma radiation on the inactivation of aflatoxin B1 in food and feed crops Efeito da radiação gama na inativação de aflatoxina B1 em alimentos e ração

    OpenAIRE

    Ghanem, I.; Orfi, M.; Shamma, M.

    2008-01-01

    Samples of food crops (peanut, peeled pistachio, unpeeled pistachio, rice, and corn) and feed (barley, bran, corn) were autoclave-sterilized, and inoculated with 10(6) of spore suspension of an isolate of Aspergillus flavus fungus known to produce aflatoxin B1 (AFB1) . Following a 10-day period of incubation at 27 C to allow for fungal growth, food and feed samples were irradiated with gamma radiation at the doses 4, 6, and 10 kGy. Results indicated that degradation of AFB1 was positively cor...

  16. Determination of aflatoxins B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil using immunoaffinity column cleanup, postcolumn derivatization, and liquid chromatography with fluorescence detection: first action 2013.05.

    Science.gov (United States)

    Bao, Lei; Liang, Chengzhu; Trucksess, Mary W; Xu, Yanli; Lv, Ning; Wu, Zhenxing; Jing, Ping; Fry, Fred S

    2013-01-01

    A collaborative study of a method for determination of aflatoxins (AFs) B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil using immunoaffinity column cleanup, postcolumn derivatization, and LC with fluorescence detection, previously published in J. AOAC Int. 95, 1689-1700 (2012), was approved as First Action 2013.05 on March 29, 2013 by the Method-Centric Committee for Aflatoxins in Edible Oils. The method uses methanol for extraction followed by filtration. The extract is applied to an immunoaffinity column with antibodies specific for AFs, which are then eluted from the column with a methanol solution. Determination and quantification occur using RP-LC with fluorescence detection after postcolumn derivatization. The average recovery of AFs in olive, peanut, and sesame oils in spiked samples (levels between 2.0 and 20.0 microg/kg) ranged from 84 to 92%. The recoveries for AFs B1, B2, G1, and G2 were 86-93, 89-95, 85-97, and 76-85%, respectively. Within-laboratory RSD (RSDr) values for AFs ranged from 3.4 to 10.2%. RSDr values forAF B1, B2, G1, and G2 were 3.5-10.9, 3.2-9.5, 6.5-14.9, and 4.8-14.2%, respectively. Between-laboratory RSD (RSDR) values for AFs were 6.1-14.5%. RSD, values for AFs B1, B2, G1, and G2 were 7.5-15.4, 7.1-14.6, 10.8-18.1, and 7.6-23.7%, respectively. Horwitz ratio values were < or =2 for the analytes in the three matrixes. PMID:24282940

  17. Rapid and quantitative detection of aflatoxin B 1 in plant grain and oilseeds products using colloid golden immuno-chromatographic method%胶体金免疫层析法快速定量分析粮油农产品中黄曲霉毒素B1

    Institute of Scientific and Technical Information of China (English)

    王督; 张文; 李培武; 张奇; 张兆威; 丁小霞; 姜俊

    2014-01-01

    采用自主研制的黄曲霉毒素B1胶体金免疫定量检测卡,建立花生、玉米、大米、小麦等粮油农产品中黄曲霉毒素B1的定量分析方法,4种样品检测的线性范围为1.0~20.0μg/kg,R2>0.97,方法的定量限为1.0μg/kg,样品加标回收率为75%~106%,RSD<20%。胶体金免疫层析法与免疫亲和柱净化-HPLC 法相比,相对误差<15%,具有简便快速、灵敏度高、重现性好等特点,适用于粮油农产品及制品中黄曲霉毒素B1筛查,样品检测时间只需15min,检测成本低于其他方法。%A rapid and quantitative detection method based on colloid gold immuno-chromatographic method was developed to analyze aflatoxin B1 in several plant samples such as peanut,corn,rice,wheat.The quantitative detection of aflatoxin B1 in these samples could be completed within 15 minutes.The dynamic linear range of the calibration curves was 1.0 μg/kg-20.0 μg/kg (R2 >0.97),and the limit of quantification was up to 1.0 μg/kg,with the recovery rate of 75%-106%(RSD<20%).Compared with immuno-affinity column purification-liquid chromatography,the relative standard deviation (RSD)was less than 15%.This detection method cost less than other methods,and with the advantage of shorter-time and higher-sensitivity,this method could be used for aflatoxin B1 screening in plant agro-products.

  18. 二氧化硅-氧化石墨烯复合物固相萃取-高效液相色谱法检测植物油中黄曲霉毒素B1、B2%Determination of Aflatoxin B1 and Aflatoxin B2 in Edible Oil by Using Graphene Oxide-SiO2 as Soild Phase Extraction Coupled with HPLC

    Institute of Scientific and Technical Information of China (English)

    王恒玲; 喻理; 李培武; 李敏; 张奇; 张文

    2014-01-01

    以二氧化硅-氧化石墨烯复合物为固相萃取材料,建立了植物油中黄曲霉毒素B1、B2的高效液相色谱(HPLC)检测方法。优化的条件为:复合材料的最佳用量为0.15 g,最佳萃取时间20 min,洗脱溶剂为乙腈,洗脱次数为2次。结果表明,在优化条件下,建立的二氧化硅-氧化石墨烯复合物固相萃取-高效液相色谱法对黄曲霉毒素B1、B2的检出限分别为0.17和0.05μg/L。将本方法应用于植物油实际样品的检测中,加标回收率在81.4%~105.3%之间,相对标准偏差为1.3%~8.6%。%Silica dioxide bound graphene oxide ( GO-SiO2 ) was applied as an effective adsorbent for determination and quantiflcation of aflatoxin B1 , B2 in edible oil by HPLC. The optimized conditions were GO-SiO2 0. 15 g, extraction time 20 min, elution reagent acetonitrile, elution cycles two times. Results showed under the optimum conditions, the detection limits of aflatoxin B1 , B2 were 0. 17 and 0. 05 μg/L, respectively. The method was successfully applied to the detection of the actual edible oil, the spiked recoveries of aflatoxin B1 and B2 were 81. 4%-105. 3% and the relative standard deviations were 1. 3%-8. 6%.

  19. 黄曲霉毒素解毒酶制剂对饲喂黄曲霉毒素B1饲粮的断奶仔猪生长性能及肝脏生化指标的影响%Effects of Aflatoxin-Detoxifizyme on Growth Performance and Hepatic Biochemical Indexes of Weaner Piglets Fed Diets Containing Aflatoxin B1

    Institute of Scientific and Technical Information of China (English)

    于会民; 梁陈冲; 陈宝江; 蔡辉益; 王勇; 刘世杰

    2013-01-01

    本试验旨在通过研究黄曲霉毒素解毒酶制剂(aflatoxin-detoxifizyme,ADTZ)对饲喂黄曲霉毒素B1(AFB1)饲粮的断奶仔猪生长性能及肝脏生化指标的影响,探讨其应用效果.选用日龄相差不超过3d、品种相同的断奶仔猪108头,按照遗传背景相同、体重相近、性别比例一致的原则随机分为3个组,分别为对照组(基础饲粮)、AFB1组(基础饲粮+0.1 mg/kg AFB1)、ADTZ组(基础饲粮+0.1 mg/kg AFB1+0.2%ADTZ),每组6个重复,每个重复6头仔猪.试验期30 d.结果表明:1)与对照组相比,AFB1组仔猪平均日增重、平均日采食量有下降趋势(P>0.05),料重比有上升趋势(P>0.05);肝脏中谷胱甘肽还原酶、过氧化氢酶、琥珀酸脱氢酶活性显著下降(P<0.05),超氧化物歧化酶、谷胱甘肽过氧化物酶与胆碱酯酶活性下降,但差异不显著(P>0.05),碱性磷酸酶活性和丙二醛含量有上升趋势(P>0.05);2)与对照组相比,当添加0.2%的ADTZ后,仔猪平均日增重、平均日采食量均有所改善,同时有降低料重比的趋势(P>0.05);以上相关生化指标均恢复到正常水平.因此,在基础饲粮中添加AFB1可导致断奶仔猪生长性能下降,肝脏生理功能受损;添加ADTZ可有效消除AFB1对断奶仔猪的危害,改善其生长性能,保护肝脏生理功能.%This experiment was conducted to evaluate the effects of aflatoxin-detoxifizyme (ADTZ) on the growth performance and hepatic biochemical indexes of weaner piglets fed diets containing aflatoxin B1 (AFB1) , and investigated the application effect. A total of 108 piglets, whose age difference was not more than three days, and with the same genetic background, body weight, and consistent sex ratio, were randomly divided into 3 groups; control group (basal diet), AFB1 group (basal diet +0.1 mg/kg AFB1) and ADTZ group (basal diet +0.1 mg/kg AFB1 +0.2% ADTZ), and with 6 replicates per group. The experiment lasted for 30 days. The results

  20. Efecto diferencial de la intoxicación crónica por aflatoxina B1 en el crecimiento y en la incidencia de lesiones hepáticas en truchas diploides y triploides (Oncorhynchus mykiss Differential effect of chronic aflatoxin B1 intoxication on the growth performance and incidence of hepatic lesions in triploid and diploid rainbow trout (Oncorhynchus mykiss

    Directory of Open Access Journals (Sweden)

    S ARANA

    2002-01-01

    Full Text Available El propósito del presente estudio fue comparar el crecimiento y la incidencia de lesiones hepáticas en truchas triploides y diploides tratadas con aflatoxina B1(AFB1. 240 truchas fueron divididas en 4 grupos: DC: truchas diploides alimentadas con ración sin AFB1; TC: truchas triploides alimentadas con ración sin AFB1; DT: truchas tratadas con ración con 80 ppb de AFB1 y TT: truchas triploides alimentadas con ración con 80 ppb de AFB1. Durante doce meses, mensualmente, cinco ejemplares de cada grupo fueron anestesiados y sacrificados. Con posteiroridad a la obtención del peso y medición del tamaño de los pesces, muestras hepáticas fueron fijadas en solución de formalina salina 10% y procesadas para análisis histopatológico. El análisis comparativo del rendimiento en crecimiento indicaron diferencias significativas entre truchas diploides del grupo control y del tratado, sugiriendo que AFB1 afecta el crecimiento de las truchas diploides. En truchas triploides no se observaron diferencias entre pesces del grupo control y los pesces tratados. El análisis histopatológico señaló que truchas triploides son más resistentes a AFB1, ya que ambos grupos tratados presentaron lesiones preneoplásicas, sin embargo, el grupo TT demostró menor incidencia de lesiones e igualmente un desarrollo más lento de las mismas. En cuanto a la ocurrencia de neoplasia, en el grupo DT 4 pesces desarrolaron carcinoma hepatocelular en el último trimestre del experimento, mientras que ningún animal triploide desarrolló lesión neoplásicaTriploid trout has been considered to be more resistant than diploid trout to many diseases and to some adverse aquaculture conditions. Considering the common problems with animal food contamination by aflatoxins, the purpose of this research was to compare the incidence of liver lesions and growth performance in triploid and diploid trout (O. mykiss exposed to chronic contamination with aflatoxin B1. A total of 240

  1. 黄曲霉毒素新型抗体制备研究进展%Research progress of aflatoxin B1 antibody preparation

    Institute of Scientific and Technical Information of China (English)

    江涛; 马良; 张宇昊; 吴春生; 蒋黎艳; 戴芳芳

    2014-01-01

    免疫学检测方法具有快捷、灵敏、特异性高的特点,在毒素的定量和定性方面已获得了快速的发展,成为毒素检测方法的研究热点。而高质量抗体的制备是建立特异性强、灵敏度高的免疫分析方法关键。目前主流研究和应用的抗体是单克隆抗体,其性质稳定,特异性强,灵敏度高。随着抗体技术的发展,重组抗体在免疫检测领域也逐渐应用。与多克隆抗体、单克隆抗体相比较而言,重组抗体具有独特的优势,可以在原核表达体系里短时间内大量生产且生产费用低廉,对黄曲霉毒素的低成本、大规模检测有重要的应用价值。本文重点阐述和分析了黄曲霉毒素单克隆抗体、重组抗体制备过程中存在的影响因素及问题,并对未来黄曲霉毒素抗体的发展前景进行了展望。%Immunological detection methods with fast, sensitive and highly specific characteristics had gained a rapid development and become a hot detection methods. High quality antibody was the key of aflatoxins determination by specific and sensitive immunoassay methods. Monoclonal antibody was the most popular antibody in current researches and commercial products for their stability, high specificity and high sensitivity. More and more recombinant antibodies were researched and applied with the development of antibody technology. Compared to these two types of antibodies, an abundance of recombinant antibody can be produced in prokaryotic expression systems in a very short time with low cost. It will be helpful to the low-cost and high throughput detection method of aflatoxins. In this paper, the preparation processing of monoclonal and recombinant antibodies in aflatoxin detection were summarized and the key factors in the preparation of aflatoxins antibody were analyzed.

  2. Determination of Aflatoxin B1 in Pharmaceutical Excipient Oil in Soft Capsules by LC-MS/MS%液-质串联法测定软胶囊中药用油辅料内黄曲霉毒素B1的含量

    Institute of Scientific and Technical Information of China (English)

    甘盛; 赖青鸟; 李志成; 韩婷; 吴超权

    2016-01-01

    目的::尝试利用液相色谱-串联质谱法对软胶囊中药用油辅料内黄曲霉毒素B1含量进行测定。方法:以甲醇-0.1%甲酸水溶液为溶剂提取软胶囊中花生油所含黄曲霉毒素B1,离心后取上清液过中性氧化铝固相萃取小柱净化,浓缩后进样测定,以甲醇-0.1%甲酸流动相梯度洗脱,流速为0.3 ml·min-1,柱温:30℃,进样量:25μl。采用电喷雾离子化四极杆串联质谱,多反应监测方式测定样品的浓度。结果:黄曲霉毒素B1在0.098~1.960μg·L-1范围内线性关系良好(r=0.9995),检出限为0.05μg·L-1,平均加样回收率为97.73%,RSD=4.625%(n=6)。结论:此法灵敏准确,专属性强,干扰少,重现性佳,对含油药物制剂中黄曲霉毒素B1含量检测有参考意义。%Objective:To assay aflatoxin B1 in the oil as a pharmaceutical excipient in soft capsules by LC-MS/MS. Methods:Aflatoxin B1 was extracted from the peanut oil in soft capsules by the solvent composed of methanol and 0. 1% formic acid solution, and then centrifuged and the supernatant was purified by neutral alumina cartridges and tested after the concentration with the mobile phase consisting of methanol and 0. 1% formic acid solution with gradient elution at the flow rate of 0. 3 ml·min-1 . 25μl of the tested solu-tion was injected for the analysis at the column temperature of 30℃. Electrospray ionization ( ESI) source was applied and operated in the position ion mode. Multiple reactions monitoring ( MRM) mode was used to quantify the samples. Results:Aflatoxin B1 was in good linearity within the range of 0. 098-1. 960 μg·L-1(r=0. 999 5). The limit of detection was 0. 05 μg·L-1. The average sampling recovery was 97. 73% (n=6) with RSD of 4. 625%. Conclusion:The method is proved to be sensitive, accurate, specified and re-producible, which is referential for the assay of aflatoxin B1 in oily preparations.

  3. 2014年黄曲霉毒素在饲料产品中的污染分布规律%Distribution laws of aflatoxin B1 contamination in feedstuffs in 2014

    Institute of Scientific and Technical Information of China (English)

    程传民; 李云; 周朝华; 柏凡; 王宇萍; 林顺全; 廖峰

    2016-01-01

    针对 1 655 个检测样品,通过酶联试剂盒初筛和液相色谱法验证两种形式进行饲料产品中黄曲霉毒素 B1污染状况调查.黄曲霉毒素B1 在雏禽配合饲料和蛋禽配合饲料中超标最严重,超标率分别为 1 1.0%和 1 4.0%.饲料产品中黄曲霉毒素B 1 污染在不同地区中存在较大差异,华中地区蛋禽配合饲料超标率达到 3 4.1%,而华北和东北地区的蛋禽配合料超标率为0.不同规模的企业饲料产品受黄曲霉毒素 B1 污染程度同样存在差异,中小企业的超标率明显高于大企业.%The aflatoxin B1 pollution status survey was studied in feedstuffs,a total of 1 655 samples were detected by ELISA initial detection and HPLC method to validation.The aflatoxin B1 in chicken feed and laying hens compound feed exceeded the target value most serious,exceeding standard rate of 11.0% and 14.0%,respectively.There was a big difference between the AFB1 pollution in different regions,in central China the exceeding standard rate was 34.1% in laying hens compound feed,but in north and northeast regions the exceeding standard rate was 0.There were also differences in the scale of different enterpri-ses,the exceeding standard rate of small and medium-sized enterprises was significantly higher than that of large enterprises.

  4. Simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 and ochratoxin A in breast milk by high-performance liquid chromatography/fluorescence after liquid-liquid extraction with low temperature purification (LLE-LTP).

    Science.gov (United States)

    Andrade, Patricia Diniz; Gomes da Silva, Julyane Laine; Caldas, Eloisa Dutra

    2013-08-23

    The aims of this study were to optimize and validate a methodology for the simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 (AFB1, AFB2, AFG1, AFG2, AFM1) and ochratoxin A (OTA) in breast milk, and to analyze these mycotoxins in samples obtained from human milk banks in the Federal District, Brazil. The optimized analytical method was based on liquid-liquid extraction with low temperature purification (3.25mL of acidified acetonitrile+0.75mL of ethyl acetate), followed by analysis by high-performance liquid chromatography with fluorescence detector (HPLC/FLD) and a photochemical post-column reactor. Limits of quantification (LOQ) ranged from 0.005 to 0.03ng/mL, recoveries from 73 to 99.5%, and relative standard deviations (RSD) from 1.8 to 17.3%. The LLE-LTP extraction method was shown to be simple and cost-effective, since no columns were needed for clean-up. Only 2 of the 224 breast milk samples analyzed were positive for the mycotoxins, both samples containing AFB2 at the LOQ level (0.005ng/mL). The identity of the mycotoxin detected was confirmed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). This result indicates that infants who are fed with breast milk from the milk banks are not at risk from aflatoxin and ochratoxin exposure.

  5. Simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 and ochratoxin A in breast milk by high-performance liquid chromatography/fluorescence after liquid-liquid extraction with low temperature purification (LLE-LTP).

    Science.gov (United States)

    Andrade, Patricia Diniz; Gomes da Silva, Julyane Laine; Caldas, Eloisa Dutra

    2013-08-23

    The aims of this study were to optimize and validate a methodology for the simultaneous analysis of aflatoxins B1, B2, G1, G2, M1 (AFB1, AFB2, AFG1, AFG2, AFM1) and ochratoxin A (OTA) in breast milk, and to analyze these mycotoxins in samples obtained from human milk banks in the Federal District, Brazil. The optimized analytical method was based on liquid-liquid extraction with low temperature purification (3.25mL of acidified acetonitrile+0.75mL of ethyl acetate), followed by analysis by high-performance liquid chromatography with fluorescence detector (HPLC/FLD) and a photochemical post-column reactor. Limits of quantification (LOQ) ranged from 0.005 to 0.03ng/mL, recoveries from 73 to 99.5%, and relative standard deviations (RSD) from 1.8 to 17.3%. The LLE-LTP extraction method was shown to be simple and cost-effective, since no columns were needed for clean-up. Only 2 of the 224 breast milk samples analyzed were positive for the mycotoxins, both samples containing AFB2 at the LOQ level (0.005ng/mL). The identity of the mycotoxin detected was confirmed by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). This result indicates that infants who are fed with breast milk from the milk banks are not at risk from aflatoxin and ochratoxin exposure. PMID:23871563

  6. Determination of Aflatoxin G2, G1, B2, and B1 in Yushangling Capsules by HPLC%高效液相色谱法测定愈伤灵胶囊中黄曲霉毒素 G2,G1,B2,B1含量

    Institute of Scientific and Technical Information of China (English)

    张正锋; 张毅

    2015-01-01

    目的:建立测定愈伤灵胶囊中黄曲霉毒素 G2,G1,B2,B1含量的高效液相色谱(HPLC)柱后光化学衍生法。方法样品经70%甲醇提取,免疫亲和柱净化,HPLC 柱后光化学衍生,荧光检测器定量,并用超高效液相色谱-串联质谱(UPLC - MS / MS)法对阳性样品进行确认。结果黄曲霉毒素 G2,G1,B2,B1含量线性范围分别是3.54~35.40 pg,7.08~70.80 pg,2.10~21.00 pg,6.24~62.40 pg( r >0.999),回收率均在70%~120%。结论该法简便、灵敏、结果准确,适用于愈伤灵胶囊中黄曲霉毒素的检测。%Objective To establish a method for determination of aflatoxin G2, G1, B2, B1 in Yushangling capsules by HPLC. Methods The samples were extracted with 70% methanol and purified with immunoaffinity column, then analyzed by HPLC with fluorescence de-tection. The positive samples were identified by ultra performance liquid chromatography - tandem mass spectrometry. Results Aflatoxin G2, G1, B2, B1 showed a good linear relationship at a range of 3. 54 - 35. 40 pg, 7. 08 - 70. 80 pg, 2. 10 - 21. 00 pg, 6. 24 - 62. 40 pg ( r > 0. 999) . The recoveries were between 70% - 120% . Conclusion The method is simple, sensitive and accurate, which is suitable for determination of four aflatoxins in Yushangling Capsules.

  7. Enzymolysis of aflatoxin B1 by Pseudomonas stutzeri F4 and analysis of the degrading products%施氏假单胞菌F4对黄曲霉毒素B1的酶解作用及其降解产物的初步分析

    Institute of Scientific and Technical Information of China (English)

    李文明; 杨文华; 李海星; 刘晓华; 曹郁生

    2013-01-01

    施氏假单胞菌F4能高效降解黄曲霉毒素B1(aflatoxin B1,AFB1).研究了F4的AFB1降解活性、降解动力学以及蛋白酶K和SDS对其降解性能的影响.F4细胞悬液与毒素共培养72 h后降解率达80.03%;蛋白酶K处理对降解率没有影响,SDS处理的细胞悬液基本丧失了降解活性.不同时间点的降解液上清液仍能有效降解残留AFB1,其中以60 h的降解液上清液活性较好,与残留AFB1继续作用48 h后降解率达84.30%;而经蛋白酶K处理后降解率仅为45.42%.低浓度AFB1诱导对菌体的降解活性没有影响.上述结果提示,F4通过胞内酶作用降解AFB1.高效液相色谱对产物分析表明,F4可将AFB1酶解为至少2种产物.%Pseudomonas stutzeri F4 is capable of removing aflatoxin B1 efficiently.The properties and the kinetics of aflatoxin B1 hydrolyzed by F4, and the influence of proteinase K and SDS on its degradation performance were investigated.After 72 h of cultivation, 80.03% of AFB1 was degraded by F4 cell suspension.Proteinase K had no affects on its degradation efficiency, while the degradation ability almost lost by SDS treatment.In addition, the cell-free supernatant of the degradation solution taken from different time also had the detoxification activity, and in which 60 h sample showed a better activity (84.30% AFB, was degraded after continuous cultivation for 48 h).When the supernatant treated with proteinase K, the degradation efficiency was decreased to 45.42%.Low concentration of AFB1 induction had no effect on the degradation activity.These results indicated that a F4 intracellular enzyme was responsible for the degradation of AFB1.Meanwhile, HPLC chromatograms demonstrated that AFB1 was degraded to at least two products.

  8. Detoxification of Aflatoxin B1 by Saccharomyces cerevisiae Mutants of Anti-Oxidative Relating Genes%酿酒酵母抗氧化相关基因突变体对黄曲霉毒素B1的清除作用

    Institute of Scientific and Technical Information of China (English)

    史锋; 黄宇啸; 李永富

    2012-01-01

    黄曲霉毒素(AF)是粮食作物和饲料原料中容易污染的一种强毒性和强致癌性物质,酿酒酵母具有毒素清除功能.利用HPLC分析了酿酒酵母野生菌BY4742及三株关键的抗氧化相关基因缺失茵zwf1△、sod2△、glr1△对黄曲霉毒素B1的清除能力.结果表明,在PBS缓冲液中存活和死亡的细胞对AFB1的清除率分别为74%~76%和71%~73%,说明酵母细胞对AFB1的清除以生物吸附作用为主.在培养基中,3种突变菌活细胞对AFB1的清除率发生不同程度的降低,其中glr1△的AFB1清除能力下降最明显,其次是sod2△,而zwf1△下降最少,说明这些关键的抗氧化基因的缺失会影响细胞在生长状态下对AFB1的清除作用.%Aflatoxins are a group of mycotoxins with strong mutagenic and carcinogenic properties. Various commodities including crop and feed materials are easy to be contaminated with aflatoxin. Saccharomyces cerevisiae have been reported to bind or degrade aflatoxin. Here, detoxification of aflatoxin B1 (AFB1 ) by wild-type strain of S. cerevisiae (BY4742) and three mutants of anti-oxidative relating genes (zw/l△, sod2△and girl △) were determined by HPLC. In PBS buffer, AFBi binding abilities of viable and dead cells were 74% -76% and 71% — 73%, respectively, indicating AFB1 was removed by yeast cells mainly through cell adsorption. In YPD medium, clearance of AFB1 by three mutant viable cells reduced, while that by wild-type BY4742 remaining high. AFB1 binding ability of g/rlA decreased most seriously, then was that of sod2△ and zwfl△. Thus, the deletion of critical anti-oxidative relating genes would decrease the AFB1 binding ability of S. cerevisiae growing cells.

  9. Comparison Experiment of Aflatoxin B1 in Inspection and Analysis%比对实验中黄曲霉毒素B1的检验与分析

    Institute of Scientific and Technical Information of China (English)

    邓建英; 孙旭峰; 凌颖敏

    2012-01-01

    本文综合国标法,色谱法及试剂盒法三种方法进行修正,把1∶1的甲醇水改为7∶3的甲醇水直接加进样品中,删除国标法及试剂盒自带方法中采用石油醚转移步骤,把手动振摇、蒸发等步骤改为漩涡振荡及离心方法,改良了样品的提取方法,操作方便,减少交叉污染,使结果更稳定.%In this paper, three methods, including GB method, chromatography method and reagent box method, were corrected for detection of Aflatoxin Blcontent The ratio of methanol to water (1: 1) was changed to 7: 3 methanol water. And the mixture was directly added into the sample. The petroleum ether-transferation step was removed from the GB and reagent kit methods. The manual shaking and evaporation were changed to vortex oscillation and centrifugal method, respectively. The sample extraction method was also improved to be more convenient,, to reduce the pollution and to make the result more stable.

  10. Determination of Aflatoxin G2, G1, B2, B1 in 34 Batches of Chinese Herbs by HPLC Associated with Post Column Photochemical Derivatization%免疫亲和柱净化HPLC柱后光化学衍生法测定34批中药材中黄曲霉毒素G2、G1、B2、B1

    Institute of Scientific and Technical Information of China (English)

    杨文武; 熊凌云; 王瑞芳; 刘岩; 孙启生; 雷雨; 王强

    2013-01-01

    目的 建立HPLC柱后光化学衍生法检测中药材中黄曲霉毒素G2、G1、B2、B1方法.方法 样品经过70%甲醇提取、免疫亲和拄净化后,采用HPLC柱后光化学衍生-荧光检测器检测中药材黄曲霉毒素含量.对疑似成分进行液质确认.结果 黄曲霉毒素G2和B2,G1和B1分别在0.75 ~ 22.5 pg和5~ 75Pg线性关系良好,方法准确稳定.检测的34批次药材中,3批酸枣仁检出黄曲霉毒素,其中1批酸枣仁黄曲霉毒素B1超过5μg·kg-1.结论 该方法简便、准确,适用于中药材黄曲霉毒素的检测.%Objective To establish HPLC methods associated with post column Photochemical Derivatization to determine aflatoxin G2, Gl, B2, Bl in Chinese herbal. Method Aflatoxins were extracted by 70% methanol and purified by an immunoaffinity column. Then the samples were analysed by HPLC fluorescence detector with post column Photochemical derivatization. Confirm the suspected components with LC-MS. Results The method with the great linear concentration range of 0.75-22.5 pg for aflatoxins G2,B2,and 5-75 pg for aflatoxins G1,B1 respectively, was stable and accurate. In the result of 34 batches of Chinese herbs,Aflatoxins were detected in 3 batches of Semen Ziziphi Spinosae among which Aflatoxin Bl of one batch exceed 5 μg·kg-l. Conclusion The method is simple and accurate,which is suitable for the determination of aflatoxins in Chinese herbal.

  11. Determination of aflatoxin B1 in propolis soft capsules by high performance liquid chromatography-tandem mass spectrometry%高效液相色谱-串联质谱法测定蜂胶软胶囊中黄曲霉毒素B1

    Institute of Scientific and Technical Information of China (English)

    苏昭仑; 叶少文; 黄康惠; 张喜金

    2015-01-01

    目的:建立测定蜂胶软胶囊中黄曲霉毒素B1的高效液相色谱-串联质谱法。方法样品经甲醇超声提取,用氰基柱将黄曲霉毒素 B1分离,用高效液相色谱-串联质谱进行测定,外标法定量。结果黄曲霉毒素 B1标准曲线在0.1~2.0 ng/mL范围内有良好的线性, r>0.9998,回收率为80.1%~87.8%, RSD0.9998), the recovery was 80.1%~87.8%, RSD<3.0%, and the limit of quantification was 1.0 μg/kg. Conclusion The established method is accurate and reliable, and it can save the pretreatment time of sample and reduce the test cost. It is suitable for the limited determination of aflatoxin B1 in propolis soft capsules of health food.

  12. 大曲中黄曲霉毒素B1提取方法的研究%Study on the Extraction of Aflatoxin B1 from Daqu

    Institute of Scientific and Technical Information of China (English)

    胡竹行; 沈才洪; 敖宗华; 王松涛; 周军; 曾娜; 丁海龙; 张强

    2014-01-01

    主要探讨了酶联免疫法检测大曲中黄曲霉毒素B1的提取条件;分别考察了超声波提取时间、超声波提取功率、甲醇浓度对黄曲霉毒素B1提取效果的影响;用正交设计找出了最佳工艺参数:甲醇浓度为55%(v/v),提取时间为25 min,提取功率为200W.采用本提取工艺所得黄曲霉毒素B1提取率明显高于国标.本研究可为大曲中黄曲霉毒素B1的检测提供简便、准确、可靠的分析方法.

  13. Degradation of Aflatoxins B1 Which in Peanut by Ammonia Gas Method%氨气熏蒸法降解花生中的黄曲霉素B1

    Institute of Scientific and Technical Information of China (English)

    赵国斌

    2014-01-01

    Peanuts which contaminated by aflatoxin was treated by ammonia gas method and AFB1 was degraded. Effect of concentration of ammonia gas, temperature, moisture of peanuts and time of degradation rate of AFB1 was investigated by single factor experiments , and the optimal condition was determined. The results show that the optimum conditions for ammonia gas method were concentration of ammonia gas 10 %, temperature 40 ℃, moisture of peanuts 30%, ammonia smoked time 96 h, the highest degradation rate 95.06%, the degradation effect was remarkable. Ammonia gas method is easy to operate, low cost, high degradation rate, and the ammonia's source is rich, it is suitable for large-scale practical application.%采用氨气熏蒸法对污染黄曲霉毒素AFB1的花生进行处理,使其中的AFB1得到降解,并通过单因素实验考察氨气浓度、氨熏温度、花生含水量和蒸熏时间等条件对降解率的影响,得到最优条件。结果表明,氨气熏蒸法最佳条件为:氨气浓度10%,熏蒸温度40℃,花生含水量30%,氨熏时间96 h,最高降解率为95.06%,降解效果显著。氨气熏蒸法易于操作、成本低廉、降解率高,且氨气来源广泛,适合大规模实际应用。

  14. 降解黄曲霉毒素B1酶的提取及降解作用研究%Study on extraction and degradation effect of enzyme which can degrade Aflatoxin B1

    Institute of Scientific and Technical Information of China (English)

    霍婷; 陶莎; 段欣; 张惠; 薛文通

    2009-01-01

    目的:从能够降解黄曲霉毒素B1的嗜麦芽窄食单胞菌中提取主要降解成分,并对其降解作用进行研究.方法:采用硫酸铵沉淀法对发酵液进行酶的提取,并对酶的提取条件进行优化实验.结果:确定种子培养基培养12h,发酵24h后,采用65%硫酸铵沉淀后的酶活性最大,对黄曲霉毒素B1的降解率达到80.25%.结论:降解黄曲霉毒素B1的菌株于40℃下种子培养12h,发酵24h,经过65%硫酸铵沉淀得到粗酶提取液,其蛋白含量为532.9μg/mL,与黄曲霉毒素B1反应72h后酶活最大,对毒素的降解率可达到81.23%,并且经过全波长扫描得到此酶在240~267nm有最大吸收峰.

  15. A reappraisal of fungi producing aflatoxins

    DEFF Research Database (Denmark)

    Varga, János; Frisvad, Jens Christian; Samson, Robert A.

    2009-01-01

    Aflatoxins are decaketide-derived secondary metabolites which are produced by a complex biosynthetic pathway. Aflatoxins are among the economically most important mycotoxins. Aflatoxin B1 exhibits hepatocarcinogenic and hepatotoxic properties, and is frequently referred to as the most potent natu...

  16. 闽南地区食源性动物组织中黄曲霉毒素B1残留量调查研究%Investigation of aflatoxin B1 residues in foodborne animal tissue in minnan region

    Institute of Scientific and Technical Information of China (English)

    朱苏琴; 张志刚; 叶葆衡; 黄印尧; 李国平

    2015-01-01

    Enzyme-linked immunosorbent (ELISA) technique was used to detect the aflatoxin B1 residual quantity in livers and mus-cles of foodborne animals which contain chicken, duck and pig in minnan region.In addition ,imported pigs were also detected.The re-suLts show that aflatoxin B1 detectable rate in pig livers and muscles were 100%,but 65% in chicken liver and 75% in duck liver, which all didn't exceed the national limited standard. The highest of AFB1 content was tested in pork liver, significantly higher than pig muscles , chicken livers and duck livers (P0.05). Besides that , AFB1 residues in imported pork though not overweight but also exist AFB1 pollution phenomenon. The resuLt of AFB1 Investigating of foodborne animal tissues suggested that problem of contamination of AFB1 has become widespread.%应用酶联免疫吸附(ELISA)技术对闽南地区食源性动物的鸡、鸭、猪肝脏和肌肉组织以及国外进口猪肌肉组织中的黄曲霉毒素B1(AFB1)残留量进行抽样检测。结果显示猪肉猪肝黄曲霉毒素B1检出率100%,鸡肝65%、鸭肝75%,但都末超过限量标率;猪肝中AFB1含量最高,显著高于猪肌肉和鸡、鸭肝中AFB1含量(P0.05);此外,国外进口猪肉中AFB1虽未超标,但也存在AFB1污染问题。调查结果表明,食源性动物组织中黄曲霉毒素B1残留问题普遍存在。

  17. Determination of Aflatoxin B1 in Liubao Tea by HPLC-Tandem Mass Spectrometry%高效液相色谱-串联质谱法测定六堡茶中黄曲霉毒素B1的研究

    Institute of Scientific and Technical Information of China (English)

    刘慧妍; 王华; 罗达龙; 陈学松

    2015-01-01

    Objective Applied in modified QuEChERS method with nano Bamboo Charcoal(NBC) as sorbents ,to establish a determination method for the Aflatoxin B1 components in Liubao tea by HPLC-Tandem Mass Spectrom-etry . Methods AFB1 in Liubao tea were extracted with acetonitrile ,The extrat was cleaned up using the mixture of NBC-PSA(primary secondary amine)-MgSO4 in dispersive solid-phase step . Results The calibration curve showed a good linearity in the range of 2.6~41.6μg/kg .The recoveries of AFB1 in Liubao tea were 83.8% ,and the relative standard deviations (RSD ,n=10) were 4.7% .The quantification (LOQ) limits were 2.6 μg/kg . Conclusion The method could meet the requirements for Aflatoxin B1 analysis in Liubao tea .%目的 建立以多壁碳纳米管作为 QuEChERS 吸附剂 ,高效液相色谱-串联质谱(UPLC-MS/MS)法测定六堡茶中黄曲霉毒素B1 的分析方法.方法 采用乙腈提取 ,并在样品提取液中加入分散固相基质净化后进行测定.结果 在 2.6 ~41.6 μg/kg浓度范围内具有良好线性关系 ,黄曲霉毒素B1 的回收率为83.8% ,相对标准偏差(RSD )4.7% ,n= 10 ,方法检测限为2.6 μg/kg.结论 该法有较好的灵敏度 ,能够满足六堡茶中黄曲霉毒素B1 测定的要求.

  18. 奶牛饲料黄曲霉毒素B1污染及控制技术研究进展%The Aflatoxin B1 Contamination in Dairy Cattle Feed and Research Progress in Control Technology

    Institute of Scientific and Technical Information of China (English)

    马美蓉; 熊江林; 傅春泉; 徐玉花

    2014-01-01

    黄曲霉毒素(AFs)具有极强毒性,在奶牛业中危害最大的是AFB1和AFM1.论文从饲料种类、季节及地域不同分析饲料黄曲霉毒素B1污染现状,并综述了物理、化学及生物等脱毒解毒技术研究进展.

  19. La exposición a la aflatoxina B1 en animales de laboratorio y su significado en la salud pública Exposure to aflatoxin B1 in experimental animals and its public health significance

    OpenAIRE

    Doralinda Guzmán de Peña

    2007-01-01

    En México se ha detectado la presencia AFB1 en humanos: como mutación en el gene p53 en hepatocarcinomas de pacientes de Monterrey, Nuevo León, México, en 1996 y como aducto AFB1-lisina en suero de pacientes del Instituto Mexicano del Seguro Social de Matamoros, Tamaulipas, México, en 2003. La aflatoxina B1 ha sido clasificada por la Agencia Internacional para Investigación en Cáncer como un agente carcinogénico para humanos. Este compuesto es un contaminante natural encontrado en alimentos y...

  20. CYP1A1 and CYP1B1 in human lymphocytes as biomarker of exposure: effect of dioxin exposure and polymorphisms

    Energy Technology Data Exchange (ETDEWEB)

    Duursen, M. van; Sanderson, T.; Berg, M. van den [Inst. for Risk Assessment Sciences, Utrecht (Netherlands)

    2004-09-15

    There are several known genetic polymorphisms of the CYP1A1 and CYP1B1 genes. A polymorphism in the 3'-untranslated region of the CYP1A1 gene (CYP1A1 MspI or CYP1A1 m1) is often studied in relation with breast or lung cancer, but little is known about the functional effect of this polymorphism. An amino acid substitution in codon 432 (Val to Leu) of the CYP1B1 gene is associated with a lower catalytic activity of the enzyme. However, the involvement of these polymorphisms on the inducibility of CYP1A1 and CYP1B1 gene expression is unclear. CYP1A1 and CYP1B1 mRNA expression levels can be determined in peripheral blood lymphocytes. This makes them potential candidates for use as biomarker of exposure to environmental compounds. Interindividual variations in mRNA expression patterns, catalytic activity and polymorphisms are very important factors when CYP1A1 and CYP1B1 expression patterns are used as biomarker of exposure, but little is known about it. Spencer et al. showed a concentration-dependent increase of CYP1B1 mRNA in lymphocytes upon exposure in vitro to 2,3,7,8-tetrachloro-p-dibenzodioxin (TCDD), the most potent dioxin. Yet, only a few studies describe the in vivo correlation between polymorphisms, mRNA expression level and exposure to environmental factors. In this study, we wanted to obtain a better insight in the CYP1A1 and CYP1B1 mRNA expression and enzyme activity in human lymphocytes. We determined the constitutive CYP1A1 and CYP1B1 mRNA expression in lymphocytes of ten healthy volunteers and the variability in sensitivity toward enzyme induction by TCDD. Further, the CYP1A1 m1 and CYP1B1 Val432Leu polymorphisms were determined.

  1. 柱前衍生高效液相色谱法测定食品中黄曲霉毒素B1、B2、G1、G2%The aflatoxin B1 ,B2 ,G1 and G2 in food were decontaminated by high performance liquid chromatograph with a sample was derived before column

    Institute of Scientific and Technical Information of China (English)

    王阳; 曹忠波

    2011-01-01

    目的:通过采用多功能小柱,利用高效液相色谱仪对食品中黄曲霉毒素B1、B2、G1、G2同时进行检测.方法:样品经体积分数为90%的乙腈溶液提取,提取液通过多功能小柱净化、浓缩,三氟乙酸(TFA)柱前衍生,C18色谱柱分离,荧光检测器检测,外标法定量.结果:4种黄曲霉毒素经过衍生后线性良好,对添加黄曲霉毒素的样品进行加标回收,回收率在80.4%~94.5%,精密度<10%.结论:该方法操作简便,线性范围广,效果良好.%Objective:The aflatoy in B1, B2, G1 and G2 in food were decontaminated by a many- sided column and detected simultaneously by high performance liquid chromatograph. Methods: The sample was extracted by acetonitrile solution with a volume fraction 90%, and then the extracted solution was decontaminated and concentrated by column, derived before trifluoroacetic acid (TFA) column, separated by C18 chromatographic column ,detected by fluorescence detector and quantified by external standard method,as seen,the four aflatoxins after derivation had good linearity. Results: The standard recovery rate of food samples with addition of aflatoxins was 80.4% ~ 94.5%, the RSD was below 10%. Conclusion: It shows the method is simple, the linear scope is wide, the result is good.

  2. Suppression of the SOS-inducing activity of Trp-P-1 and aflatoxin B1 by meso-dihydroguaiaretic acid from Machilus thunbergii in the Salmonella typhimurium TA1535/pSK1002 umu test.

    Science.gov (United States)

    Miyazawa, M; Okuno, Y; Oshiro, K; Kasahara, H; Shimamura, H; Nakamura, S; Kameoka, H

    1998-07-01

    The methanol extract from Machilus thunbergii showed a suppressive effect on umu gene expression of the SOS response in Salmonella typhimurium TA1535/pSK1002 against the mutagen, 3-amino-1,4-dimethyl-5H-pyrido[4,3-b]indole (Trp-P-1), which requires liver metabolizing enzymes. The methanol extract from M. thunbergii was successively re-extracted with chloroform, butanol and water. A suppressive compound in the chloroform extract fraction was isolated by SiO2 column chromatography and identified as meso-dihydroguaiaretic acid by GC-MS, and 1H- and 13C-NMR spectroscopy. Meso-dihydroguaiaretic acid inhibited of the SOS-inducing activity of Trp-P-1 in the umu test. Gene expression was suppressed by 62% at less than 0.18 mumol/ml, the ID50 value being 0.08 mumol/ml. Compound 1 was also assayed with aflatoxin B1 (AfB1) and showed a suppressive effect. PMID:9720227

  3. Aflatoxin in Tunisian aleppo pine nuts.

    Science.gov (United States)

    Boutrif, E; Jemmali, M; Pohland, A E; Campbell, A D

    1977-05-01

    Twenty-six of 50 Aleppo pine nuts samples collected throughout Tunisia showed relatively high levels of contamination by aflatoxin. Some samples contained as much as 2000 ppb aflatoxin B1, and very few contained less than 100 ppb. Total aflatoxins as high as 7550 ppb were found. A traditional pudding, widely consumed in Tunisia, which was prepared from contaminated nuts still contained more than 80% of the aflatoxin originally present in the nuts.

  4. To Optimize the Extraction Method of Aflatoxin B1 from Naturally Contaminated Rice%大米中黄曲霉毒素B1的提取方法优化

    Institute of Scientific and Technical Information of China (English)

    孙秀兰; 赵晓联; 汤坚

    2004-01-01

    通过ELISA检测,对多种溶剂提取大米中黄曲霉毒素B1的方法进行了优化,结果显示对于不同污染程度的大米样品,50%丙酮/水溶液和50%甲醇/水溶液提取效率最高,0.1、1、10μg/kg三个浓度水平的加标回收率为80.4%~140%,方法的重复性和稳定性很好,相对标准偏差2.99%~9.61%.

  5. Amperometric biosensor based on acetylcholinesterase for the detection of aflatoxin B1%基于乙酰胆碱酯酶传感器的黄曲霉毒素B1检测方法研究

    Institute of Scientific and Technical Information of China (English)

    袁蓓; 赵凤娟; 卫敏

    2016-01-01

    基于黄曲霉毒素B1(AFB1)对乙酰胆碱酯酶(AChE)的抑制作用,建立了抑制型酶传感器快速检测AFB1的方法.对实验参数进行优化,得到PBS缓冲溶液的最优pH为7.3、戊二醛的最佳质量分数为0.5%、最佳抑制时间为1 8 min.在最优条件下,建立标准曲线,并对样品进行检测.该方法对AFB1的检测线性范围为3.00~14.00 μg/mL,线性方程为y=2.509x+13.857(R2=0.994),检出限为0.86μg/mL,花生油中AFB1的加标回收率为87.49%~ 109.51%,玉米中AFB1的加标回收率为89.00%~105.50%.

  6. 黄曲霉毒素B1(AFB1)降解菌的筛选鉴定%Screening and Identification of Aflatoxin B1(AFB1)Degradation Strains

    Institute of Scientific and Technical Information of China (English)

    陈晓飞; 周伏忠; 孙玉飞; 宁萌; 常丽

    2011-01-01

    Using cumarin plate as the priliminaiy screening procedure,then co-culture and AFB, degradation test as the second screening step of the strains. It was found that four strains of which have the AFB1 degradation ability: the degradation ratio of AFB1 of G3-21 and Tl-4-1 reached 65 %.the degradation ratio of AFB1 of the other two strains Y2-30 and Tl-3-7 reached 45 %. From the analysis of colony morphology and 16S rDNA gene sequence,the strains G3-21 and Y2-30 were finally identified as Pseudomonas putida. The strain Tl-4-lwas Streptomyces sp. The strain Tl-3-7was Firmicutes bacterium.%以香豆素为唯一碳源和能源的培养基进行初筛,然后将菌株与黄曲霉毒素B1(AFB1)共培养后测定AFB1降解率的方法进行复筛,最后筛选出4株高活性AFB,降解菌;其中G3-21和T1-4-1对AFB1降解率可达到65%,Y2-30和T1-3-7对AFB1的降解率达到45%以上,形态和16S rDNA序列分析鉴定结果表明,G3-21和Y2-30为恶臭假单胞菌(Pseudomonas putida),T1-4-1为链霉菌(Streptomyces sp.),T1-3-7为厚壁菌(Firmicutes bacterium).

  7. 免疫亲和柱净化-柱后衍生高效液相色谱荧光法快速检测玉米粉中黄曲霉毒素B1%Determination of Aflatoxin B 1 in Corn Flour by Immune Affinity Column Purification and HPLC-FLD with Post-Column Photochemical Derviatization

    Institute of Scientific and Technical Information of China (English)

    郑睿行; 韩丹; 祝华明; 王怡念; 郑微波; 鲍利锋

    2014-01-01

    本文建立了免疫亲和柱净化-高效液相色谱荧光法快速检测玉米粉中黄曲霉毒素B1aflatoxin B1, AFB1)的方法。样品经甲醇/水(4∶1, v/v)浸提,免疫亲和柱净化,高效液相色谱分离,光化学柱后衍生,荧光检测器测定。结果表明: AFB1在2.5~150μg/L 具有良好的线性关系,相关系数为R2=0.9998,检测限为1.0μg/kg,平均回收率为84.0%~96.3%, RSD范围为0.7%~2.5%。该方法具有快速、准确和重复性好的优点,适用于大批次玉米及玉米粉制品中AFB1的检测。%A method for determination of aflatoxin B1 ( AFB1) in corn flour using immune-affinity column purification and HPLC with post column photo-chemical derivatization was developed. Samples were extracted with methanol-water solution (4∶1, v/v), and purified by immune-affinity columns. Seperation of AFB1 were conducted by HPLC and determination was carried out by fluorescence detector after photochemical derivatization. Results showed that this method had good linearity range from 2.5-150 μg/L, R2=0.9998, detection limit was 1.0μg/kg, recoveries of analytes were from 84.0%~96.3%, RSD was 0.7%~2.5%. Method is quick, precise, high sensitive and good repeatability, which is suitable for the determination of AFB1 in corn flour.

  8. Effects of Glucose Oxidase on Growth Performance, Serum Parameters and Slaughter Performance of Meat Ducks and Its Antidotal Effect on Aflatoxin B1%葡萄糖氧化酶对肉鸭生长性能、血清指标和屠宰性能的影响及其解除黄曲霉毒素 B1效果

    Institute of Scientific and Technical Information of China (English)

    汤海鸥; 高秀华; 姚斌; 张广民; 王振兴; 李晓存

    2015-01-01

    This experiment was conducted to investigate the effects of diets supplemented with glucose oxidase ( GOD) on growth performance, serum parameters and slaughter performance of meat ducks and its antidotal effect on aflatoxin B1. A total of 100 Cherry Valley ducks (1-day-old) were randomly assigned to five groups with four replicates of fifty ducks in each replicate. The control group ( groupⅠ) was fed on a basic diet. The two test groups ( groupsⅡandⅢ) were fed on basic diet supplelemted with 100 and 200 g/t GOD. Aflatoxin B1 control group ( groupⅣ) was fed on basic diet with moldy corn. The groupⅤwas fed on the contaminated diet with 200 g/t GOD. The experiment lasted for 46 days. The results showed as follows:1) the average dai-ly gain and ratio of feed to gain of ducks in groupsⅡ andⅢ were significantly better than those in the control group ( P0.05) . The experiment indicated that the growth performance of ducks fed on basic diet can be significantly im-proved by supplementing GOD. The growth performance, serum parameters, slaughter performance and mor-tality rate of ducks fed diet with aflatoxin B1 can be improved by supplementing GOD.%本试验旨在研究饲粮中添加葡萄糖氧化酶( GOD)对肉鸭生长性能、血清指标和屠宰性能的影响及其解除黄曲霉毒素B1(AFB1)的效果。选用1000只健康的1日龄樱桃谷鸭,随机分为5个组,每组4个重复,每个重复50只鸭。Ⅰ组为对照组,饲喂基础饲粮;Ⅱ、Ⅲ组分别在基础饲粮中添加100和200 g/t GOD;Ⅳ组为混有AFB1超标玉米的攻毒组;Ⅴ组在Ⅳ组饲粮中添加200 g/t GOD。试验期46 d。结果表明:1)与对照组相比,Ⅱ、Ⅲ组显著提高了平均日增重( P0.05),显著低于其他各组(P<0.05)。由此可见,常规饲粮中添加GOD能显著改善肉鸭生长性能,添加GOD可改善AFB1攻毒肉鸭的生长性能、屠宰性能和血清指标,降低死亡率。

  9. 电化学免疫传感器检测样品中黄曲霉毒素B1的误差分析%Study on error analysis during detecting aflatoxin B1 in food by using electrochemical immunosensor

    Institute of Scientific and Technical Information of China (English)

    冯甜; 张弦; 杨弦弦; 李朝睿

    2014-01-01

    目的:探讨扩大电化学免疫传感器在检测食品及饲料中黄曲霉毒素B1(A FB1)的应用范围,分析抗体孵育时间及各种前处理方法对检测结果的影响。方法利用A FB1和羧基化单壁碳纳米管构建的双层免疫传感器,采用循环伏安法对传感器进行验证,研究了抗体孵育时间和样品前处理方法对检测结果的影响。结果 AFB1抗体和二抗的最佳孵育时间为90 min ,且不同的前处理方法对检测结果影响较大,粗提溶液经过A FB1亲和柱纯化、浓缩能够有效排除样品基质干扰物对测定结果的影响。结论电化学免疫传感器检测A FB1具有快速、简便、检测限低等特点,其稳定性受到抗体孵育时间以及样品前处理等因素影响。%Objective To expand the application of electrochemical immunosensor during deleting aflatoxin B1 in foods and feeds through analyzing impacts of the time of antibody incubation and sample preparation .Methods T he double self-assembly immu-nosensor combined with aflatoxin B1 and carboxylated single-walled carbon nanotubes (SWNTs) was characterized by cyclic volta-mmetry and impacts of the time of antibody incubation and sample preparation methods were investigated .Results The signal in-creased gradually following the increasing time of antibody incubation and reached a plateau at 90 min and sample preparation meth-ods showed a comparatively large impact on results .Additionally ,the crude extractions purified through removing interfering com-pounds by immunoaffinity column could effectively eliminate the interference effects of sample matrix .Conclusion Deleting aflatox-in B1 by electrochemical immunosensor is characterized by various features ,such as fast ,simple and low detection limits .The pres-ent study shows that stability of the electrochemical immunosensor is affected by the time of antibody incubation and sample prepa-ration .

  10. Preparation and Application of Graphene/Conductive Polymer/Ionic Liquid Immunosensor for Determination of Aflatoxin B1%石墨烯/导电高分子/离子液体修饰的黄曲霉毒素B1免疫传感器的制备及应用

    Institute of Scientific and Technical Information of China (English)

    周琳婷; 李在均; 方银军

    2012-01-01

    依次电沉积氧化石墨烯、2,5-二(2噻吩)-1-对苯甲酸吡咯和氯金酸于金电极表面,以EDC/NHS为活化剂,将黄曲霉毒素B1(AFB1)抗体共价连接在导电高分子膜上,最后滴涂1,3-二丁基咪唑六氟磷酸盐离子液体于上述修饰电极表面,制得AFB1免疫传感器.以Fe(CN)63-/4-的磷酸盐缓冲溶液(pH 7.0)为测试底液,采用循环伏安法和交流阻抗法考察此免疫传感器的电化学行为.研究表明:石墨烯和纳米金的引入明显提高了修饰层的电子转移速率,使电极的表观活性面积由裸金电极的0.1772 cm2增加到0.2188 cm2和0.2640 cm2.当AFB1浓度在3.2×10-15~3.2×1013mol/L范围内,传感器的交流阻抗响应值与浓度呈线性关系,相关系数R2=0.994,检出限为11×10-15 mol/L.传感器在4℃下保存20周以上,电化学响应保持基本不变.本方法的灵敏度和稳定性优于现有文献报道,并应用于花生样品中痕量AFB1的测定.%To fabricate the aflatoxin B1 immunosensor, graphene oxide, 2, 5-di-(2-thienyl)-l-pyrrole-l-(p-benzoic acid) and chloroauric acid were electrodeposited on the gold electrode surface in sequence. Then aflatoxin B1 (AFB1) antibody was covalently connected to the conducting polymer film with 1-ethyl-(3-dimethyl-aminopropyl)-carbodimide/N-hydroxysuccimimide (EHC/NHS) as activator and 1,3-dibutyliminazolium hexafluorophosphate ionic liquid was finally coated on the modified electrode. The Fe(CN)63-/4-phosphate buffer solution (pH 7. 0) was employed as base solution for investigating electrochemical performances of the immunosensor by cyclic voltammetry and electrochemical impedance spectroscopy. Research revealed the introduction of graphene and gold nanocomposite obviously improved electron transfer rate of the modified layer, and the apparent electroactive surface areas of the electrode also increased up to 0. 2188 cm2 and 0. 2640 cm2 from 0. 1772 cm2 of bare gold electrode, respectively. When aflatoxin B1 concentration

  11. Rapid Determination of Aflatoxin B1,B2,G1,G2 in wine with HPLC and Immunoaffinity Column%免疫亲和柱HPLC荧光检测酒中黄曲霉毒素B1、B2、G1、G2

    Institute of Scientific and Technical Information of China (English)

    李佐卿; 谢东华; 孙大为; 康继韬; 俞雪钧

    2001-01-01

    Monoclonal antibody immunoaffinity technology was used for specific method of extracting and purging aflatoxin from samples directly. After evaporating the extract to dryness, the residue were derived and detected by HPLC fluorescence detector. The average recoveries (n=10) are G1 73.8%、B1 97.3%、G2 61.7%、B2 90.5% respectively;the RSD of ten determination are G1 4.50%、B1 3.80%、G2 3.68%、B2 4.77% respectively; in the rang of 25—1250pg the analytical response is linear, the correlation coefficients are G1: r =0.9990 ,B1: r=0.9994,G2: r=0.9995,B2 :r=0.9992 respectively;the limit of determination was 6.25pg .%采用单克隆抗体免疫亲和技术作为直接从样品中分离提纯黄曲霉毒素的特效手段,提取液挥发干后,经衍生用HPLC荧光检测器测定。本法在样品中添加2.5μg /kg黄曲霉毒素时进行10次测定,平均回收率分别为G1 73.8%、B1 97.3%、G2 61.7%、B2 90.5%;2.5μg /kg 10次测定的精密度分别为:G1 4.50%、B1 3.80%、G2 3.68%、B2 4.77%,本方法在25—1250pg 范围内呈线性,相关系数分别为G1: r =0.9990 、 B1: r=0.9994、 G2: r=0.9995、 B2 : r=0.9992。测定的最低检出限为6.25pg。

  12. Characterization of the rat glutathione S-transferase Yc2 subunit gene, GSTA5: identification of a putative antioxidant-responsive element in the 5'-flanking region of rat GSTA5 that may mediate chemoprotection against aflatoxin B1.

    Science.gov (United States)

    Pulford, D J; Hayes, J D

    1996-08-15

    We have isolated and characterized genomic DNA encoding the rat glutathione S-transferase Yc2 subunit. This protein is now referred to as rGSTA5 and is noteworthy because of its high activity towards aflatoxin B1-8,9-epoxide, its marked inducibility by chemoprotectors, its sex-specific regulation, and its over-expression in hepatoma and preneoplastic nodules. The rGSTA5 gene, which was isolated on two overlapping bacteriophage lambda clones, is approx. 12 kb in length and, unlike other class Alpha genes described to date, it comprises six exons. The transcription start site has been identified 228 bp upstream from the ATG translational initiation codon, and is situated 51 bp downstream from a consensus TATA-box. Deletion analysis, using luciferase reporter constructs, has shown that the region between -177 bp and +65 bp from the transcriptional start site contains a functional promoter. Computer-assisted analysis of the upstream sequence has indicated the presence of an antioxidant-responsive element (ARE), and several elements thought to be required for tissue-specific expression of the enzyme. In addition, several putative oestrogen-responsive half sites were observed in both upstream and intronic sequences. PMID:8761455

  13. Efficacy of aqueous garlic extract on growth, aflatoxin B1 production, and cyto-morphological aberrations of Aspergillus flavus, causing human ophthalmic infection: topical treatment of A. flavus keratitis.

    Science.gov (United States)

    Ismaiel, Ahmed A; Rabie, Gamal H; Kenawey, Saied E M; Abd El-Aal, Marwa A

    2012-10-01

    By using agar well diffusion assay, antifungal activity of aqueous extract prepared from Egyptian garlic (Allium sativum L.) was evaluated in vitro against two strains of Aspergillus flavus (OC1 and OC10) causing human ocular infection. The recorded minimum inhibitory concentration (MIC) for growth inhibition of both strains was 3.60 mg/ml. Aqueous garlic extract (AGE) was used in successive in vivo tests as an attempt to cure rabbit's fungal keratitis caused by A. flavus OC1. Findings showed that diluted preparation of AGE was effective topical antifungal agent and succeeded to cure severe A. flavus keratitis in a time course less than 10 days without any observable side effects. Microscopic examination showed that AGE induced deleterious cyto-morphological aberrations in A. flavus target cells. AGE applied to Czapek's broth via contact method was more effective on growth, spores and aflatoxin B1 production than AGE applied to the same broth at the same concentration via fumigation method. PMID:24031964

  14. A fabricated electro-spun sensor based on Lake Red C pigments doped into PAN (polyacrylonitrile) nano-fibers for electrochemical detection of Aflatoxin B1 in poultry feed and serum samples.

    Science.gov (United States)

    Babakhanian, Arash; Momeneh, Tahereh; Aberoomand-azar, Parviz; Kaki, Samineh; Torki, Mehran; Hossein Kiaie, Seyed; Sadeghi, Ehsan; Dabirian, Farzad

    2015-11-21

    The aim of this work was to fabricate a novel nano-fiber modified electrode, involving Lake Red C (LRC) pigments doped into electrospun polyacrylonitrile (PAN) fibrous films. Cyclic voltammetry (CV), scanning electron microscopy (SEM), electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV) techniques were used for electrochemical and morphological characterization of the composite fibers. This sensor responds to Aflatoxin B1 (AFB1) over the concentration range of 40-120 nM with high accuracy and precision in analysis. The modified electrode exhibited an excellent electrocatalytic ability (α = 0.42, log K(s) = 4.21 s(-1), and Γ = 1.49 × 10(-5) mmol cm(-2)) for reduction of AFB1 at the optimum pH of 6 and working potential of -0.75 V (vs. SCE). The common substances accompanying AFB1 had no serious interferences on the response of the modified electrode to AFB1. The modified electrode indicated reproducible behavior and a high level stability during the experiments, making it particularly suitable for the analytical determination of AFB1 in poultry feed and serum samples. PMID:26460282

  15. Determination of Aflatoxins in Taditional Chinese Medicine by Cleaning up the Immunoaffinity Column and HPLC with Post Column Photochemical Derivatization%免疫亲和柱-HPLC柱后光化学衍生法测定中药饮片中黄曲霉毒素B1、B2、G1、G2的含量

    Institute of Scientific and Technical Information of China (English)

    朱迪; 谭丹; 向文英; 孙佳; 李勇军; 王爱民

    2015-01-01

    目的:建立中药饮片中黄曲霉毒素( AF)B1、B2、G1、G2含量测定的方法。方法:收集中药饮片87批,用免疫亲和柱进行AF的分离处理,采用高液相色谱( HPLC)柱后光化学衍生化法测定中药饮片中AFB1、AFB2、AFG1、AFG2含量并进行方法学分析。结果:被测定的AFB1、AFB2、AFG1、AFG2的检出限分别为0.7、0.3、0.6和0.3μg/kg,在选定的范围内线性关系良好( r≥0.999),精密度试验、重复性试验、准确度试验及标准样品验证试验结果均良好,87批中药饮片均未检出黄曲霉毒素。结论:免疫亲和柱-HPLC柱后光化学衍生化法操作简便,专属性强、灵敏度高,检测中药饮片中AFB1、AFB2、AFG1及AFG2的含量准确可靠。%Objective:To establish a method for determination of aflatoxins in Chinese medicinal de-coction pieces and to promote quality control of Chinese medicinal decoction pieces. Methods:Eighty-seven batches of Chinese medicinal decoction pieces were collected and immunoaffinity column and HPLC with post column photochemical derivatization was adopted to measure aflatoxins content,inclu-ding aflatoxin B1,aflatoxin B2,aflatoxin G1,aflatoxin G2 in all samples. Results:The detection lim-it of aflatoxin B1 ,aflatoxin B2 ,aflatoxin G1 ,aflatoxin G2 was 0 . 7 ,0 . 3 ,0 . 6 and 0 . 3 μg/kg respec-tively. There was a a good linear relationship(r≥0. 999)in selected range of content. The results of precision test,repeatability test,accuracy test were good as well. In 87 different batches of Chinese medicinal decoction pieces,aflatoxins were not detected. Conclusion:The mothod of Immunoaffinity column and HPLC with post column photochemical derivatization is convenient,specific,sensitive,ac-curate and reliable in measuring aflatoxins content of Chinese medicinal decoction pieces.

  16. HPLC-柱后光化学衍生法检测花生酱中黄曲霉毒素%Determination of Aflatoxin B1,B2,G1 and G2 in Peanut Butter by HPLC with On-Line Post-Column Photochemical Derivatization

    Institute of Scientific and Technical Information of China (English)

    邵丽; 王晓; 滕振勇

    2015-01-01

    To determine the contents of aflatoxin B1,B2,G1 and G2 in peanut butter using on-line post-column photo-chemical derivatization-HPLC-FLD method. The samples were extracted with acetonitril-H2O (80:20) and purified with inmunoafinity column,aflatoxins were analyzed by HPLC -FLD with post -column photochemical derivatizaton. On optimum conditions,aflatoxin B1,G1 ranging 0.30 mg/L-10 mg/L showed a good linear relationship with aflatoxin B2,G2 ranging 0.06 mg/L-3.0mg/L with r>0.998. The recoveries ranged between 80 % and 101 % ,with RSDs all bellow 5.9 %. LOD of aflatoxin B1,B2,G1 and G2 were 0.10,0.03,0.15,0.04μg/kg,respectively.%建立高效液相色谱-在线柱后光化学衍生-荧光检测器检测花生酱中黄曲霉毒素B1、B2、G1、G2的含量.样品以乙腈-水(80:20)溶液提取,经免疫亲和柱净化后,利用在线柱后光化学衍生-HPLC-FLD进行分析测定.结果:在优化条件下,黄曲霉毒素B1、G1在0.30 mg/L~10 mg/L,黄曲霉毒素B2、G2在0.06 mg/L~3.0mg/L线性关系良好,r>0.998 ,回收率80%~101%,RSD<5.9%.黄曲霉毒素B1、B2、G1、G2的检测限(LOD)分别为0.10、0.03、0.15、0.04μg/kg.

  17. 高效液相色谱-串联质谱法同时测定花生制品中的黄曲霉毒素B1、B2、G1、G2%Simultaneous Determination of Aflatoxin B1, B2, G1 and G2 in Peanuts by HPLC-MS

    Institute of Scientific and Technical Information of China (English)

    冯伟科; 罗佳玲; 赖毅东

    2011-01-01

    采用高效液相色谱-串联质谱建立了高效液相色谱-串联质谱法同时测定花生制品中4种黄曲霉毒素(B1、B2、G1、G2)的方法.结果表明,在ESI正离子模式下,高效液相色谱-串联质谱法的最低检出限为0.2μg/kg,定量限为0.6 μg/kg;标准工作液在0.5~50.0μg/kg的范围内线性良好,相关系数达到0.9990.%Liquid chromatography-tandem mass spectrometry was used for the simultaneous quantitative analysis of aflatoxin B1, B2, G1 and G2 in peanuts. The eluted extract was analyzed by HPLC-MS/MS in positive ion mode using multiple reactions monitoring with a triple-quadruple MS using an electrospray ionization source. The limits of detection (LODs, S/N=3) and the limits of quantification for the target compounds were in the range of 0.2 and 0.6 μg/kg, respectively. The calibration curve was linear in the concentration range of 0.5~50 μg/kg with the regression coefficient of 0.9990.

  18. Evaluation of the Uncertainties in the Determination of Aflatoxins B1 in Dry Cassava by HPLC%高效液相色谱法测定木薯干中黄曲霉毒素B1的不确定度评定

    Institute of Scientific and Technical Information of China (English)

    宋卫得; 厉雪芹; 高尧华; 刘培海; 刘冰; 卢志晓; 孙灿

    2016-01-01

    The uncertainties in the determination of aflatoxins B1 in dry cassava by HPLC according to GB/T 18979-2003 were evaluated. The uncertainty components were classified, the relative standard uncertainty of each component was calculated, and the combined standard uncer-tainty and expanded uncertainty were calculated eventually. The evaluation results suggested that the component with the maximum uncertainty influence was recovery rate and the component with the minimum uncertainty influence was sample weighing, with the relative combined stan-dard uncertainty of 10.5%.%依据GB/T18979—2003《食品中黄曲霉毒素的测定》,对免疫亲和层洗净化高效液相色谱法测定木薯干中黄曲霉毒素B1的不确定度进行了评定.对检测过程中产生的不确定分量进行了分类,计算各分量的相对标准不确定度,从而计算出合成标准不确定度和扩展不确定度.通过评估发现,对测定不确定度影响最大的分量是检测方法的回收率,影响最小的分量是样品质量的称量,合成相对标准不确定度为10.5%.

  19. Sensitive quantification of aflatoxin B1 in animal feeds, corn feed grain, and yellow corn meal using immunomagnetic bead-based recovery and real-time immunoquantitative-PCR.

    Science.gov (United States)

    Babu, Dinesh; Muriana, Peter M

    2014-01-01

    Aflatoxins are considered unavoidable natural mycotoxins encountered in foods, animal feeds, and feed grains. In this study, we demonstrate the application of our recently developed real-time immunoquantitative PCR (RT iq-PCR) assay for sensitive detection and quantification of aflatoxins in poultry feed, two types of dairy feed (1 and 2), horse feed, whole kernel corn feed grains, and retail yellow ground corn meal. Upon testing methanol/water (60:40) extractions of the above samples using competitive direct enzyme linked immunosorbent assay, the aflatoxin content was found to be animal and human health, the RT iq-PCR method described in this study can be useful for quantifying low natural aflatoxin levels in complex matrices of food or animal feed samples without the requirement of extra sample cleanup.

  20. QuEChERS-酶联免疫快速检测法测定茶叶中黄曲霉毒素B1%Simultaneous determination for aflatoxin B1 in tea leaf by QuEChERS -enzyme-linked immunosorbent assay

    Institute of Scientific and Technical Information of China (English)

    刘辉; 张燕

    2015-01-01

    目的:建立QuEChERS-酶联免疫快速检测茶叶中黄曲霉毒素B1的方法,并对样品前处理条件进行优化。方法茶叶样品用70%乙腈水提取溶液进行提取目标物,离心后取上清液并用PSA+MgSO4进行净化处理后,采用酶联免疫快速检测方法进行分析测定。结果本方法的检出限为0.078µg/kg,线性范围为0.125~0.854µg/kg, IC50值为0.327 ng/mL。在三个不同添加水平下,样品的平均回收率为87.66%~97.17%,相对标准偏差为4.89%~7.16%。检测结果与高效液相色谱法(high performance liquid chromatography, HPLC)方法的相关系r2=0.9854,线性相关性良好。结论该方法更加简便、快速、高效,能够用于茶叶中黄曲霉毒素B1的检测。%Objective To establish a QuEChERS-enzyme-linked immunosorbent assay (ELISA) method for determining the aflatoxin B1 (AFB1) in tea leaf rapidly, and optimizing the sample pre-treatment conditions. Methods The samples were extracted with 70% acetonitrile in water, and the supernatant liquid was directly purified with PSA+MgSO4 for detection of ELISA.Results The limit of detection (LOD) of method was 0.078 µg/kg, with linear range between 0.125 and 0.854 µg/kg, and IC50 value was 0.327 ng/mL. Average recovery rate of samples was between 87.66% and 97.17% at 3 adding levels, and the relative standard deviation (RSD) was 4.89%~7.16%. The results showed a good coefficient with high performance liquid chromatography (HPLC) (r2=0.9854).Conclusion This method proved to be suitable for the screening of tea leaf samples for the presence of AFB1, which was easier, faster and more efficient.

  1. 免疫亲和柱净化-在线柱后光化学衍生-HPLC-FLD同时测定甘草中黄曲霉毒素B1,B2,G1,G2和赭曲霉毒素A的含量%Simultaneous determination of aflatoxin B1, B2, G1, G2 and ochratoxin A in Glycyrrhiza uralensis by HPLC-FLD after immunoaffinity column with online post-column photochemicai derivatization

    Institute of Scientific and Technical Information of China (English)

    韦日伟; 杨小丽; 仇峰; 杨美华; 覃洁萍

    2011-01-01

    目的:建立同时检测甘草中黄曲霉毒素B1,B2,G1,G2和赭曲霉毒素A的免疫亲和柱净化-在线柱后光化学衍生-HPLC- FLD测定的方法.方法:样品经甲醇-水(80:20)超声提取后,用免疫亲和柱净化和富集;以甲醇和0.5%乙酸溶液为流动相进行梯度洗脱,通过柱后光化学衍生,荧光检测器测定.结果:黄曲霉毒素G2,G1,B2,B1和赭曲霉毒素A的检测限分别为0.02,0.06,0.015,0.03,0.25μg·kg-1,平均加样回收率为76.0%~103%,RSD低于13%.结论:该方法快速简便、准确,可用于甘草中同时测定黄曲霉毒素B1,B2,G1,G2和赭曲霉毒素A的含量.%Objective: To develop a method for the simultaneous determination of aflatoxin B1, B2, G1, G2 and ochratoxin A in Glycyrrhita walensis by HPLC-FLD after immunoaffinity column with online post-column photochemical derivatization. Method; Sample was extracted with MeOH:- H2O (80:20) and cleaned up by immunoaffinity column. The toxins were separated by reversed-phase HPLC and the mobile phase was consisted of methanol and 0. 5% acetic acid solution with gradient elution. The determination was carried out by fluorescence detector after photochemical derivatization. Result; The detection limits of aflatoxin G2 , C,, B2 , B, and ochratoxin A were 0.02, 0. 06, 0. 015, 0.03 and 0. 25 μg · kg-1, respectively. The recoveries of analytes were from 76.0% to 103% and the relative standard deviations (RSDs) were below 13%. Conclusion; The method is a simple, accurate and can be used to determine the contents of aflatoxin B1, B2, G1, G2 and ochratoxin A in G. Uralensis simultaneously.

  2. Inhibition of aflatoxin production by selected insecticides.

    OpenAIRE

    Draughon, F A; Ayres, J. C.

    1981-01-01

    The insecticide naled completed inhibition production of aflatoxins B1, B2, G1, and G2 by and growth of Aspergillus parasiticus at a 100-ppm (100 microgram/ml) concentration. The insecticides dichlorvos, Landrin, pyrethrum, Sevin, malathion, and Diazinon significantly (P = 0.05) inhibited production of aflatoxins at a 100-ppm concentration. However, at a concentration of 10 ppm, significant inhibition in production of aflatoxins was found only with naled, dichlorvos, Sevin, Landrin, and pyret...

  3. 高效液相色谱-串联质谱法同时测定地龙中4个黄曲霉毒素%HPLC-MS/MS simultaneous determination of aflatoxins B1 , B2 , G1 and G2 in Pheretima

    Institute of Scientific and Technical Information of China (English)

    杨小丽; 仇峰; 韦日伟; 覃禹; 杨美华; 欧阳臻

    2012-01-01

    Objective:To establish a sensitive and accurate liquid chromatography -tandem mass spectrometry method(LC -MS/MS) for simultaneous determination of four aflatoxins in Pheretima. Methods;The samples were firstly extracted with methanol - water solution ( 80:20, v/v), and then cleaned up by immunoaffinity columns. The mass spectrometer was operated in the positive ionization electrospray( ESI) mode using multiple reaction monitoring (MRM) for analysis of four aflatoxins. The transitions of m/z 313-241 (aflatoxin B, ,CE 50 eV) ,m/z 315-259 (aflatoxin B2,CE 43 eV) ,m/z 329-243 (aflatoxin G1,CE 38 eV) and m/z 331-245(aflatoxin G2,CE 40 eV) were used to quantify four aflatoxins,respectively. Results:The detection limits of aflatoxin B1,B2 ,G1 and G2 were 0. 03,0. 02,0. 03 and 0. 02 μg o kg-1 .respectively. The recoveries of four analytes ranged from 88. 0% to 100. 3% and the relative standard deviations were all below 6. 1 %. Conclusion; The method is sensitive, simple and accurate, and proved to be suitable for the simultaneous determination of four aflatoxins in Pheretima.%目的:建立同时测定地龙中4个黄曲霉毒素含量的高效液相色谱串联质谱法.方法:样品经甲醇-水(80∶20,v/v)提取,通过免疫亲和柱净化后,采用高效液相色谱串联质谱法测定其中4个黄曲霉毒素的含量,以多反应监测(MRM)方式分别监测离子对m/z 313→241(黄曲霉毒素B1,CE 50 eV),m/z 315→259(黄曲霉毒素B2,CE 43 eV),m/z329→243(黄曲霉毒素G1,CE 38 eV)和m/z 331→245(黄曲霉毒素G2,CE 40 eV).结果:黄曲霉毒素B1、B2、G1、G2的检测限分别为0.03,0.02,0.03,0.02μg·kg-1,回收率在88.0%~100.3%范围内,RSD均低于6.1%.结论:该方法快速、灵敏,结果准确,适用于地龙中4个黄曲霉毒素的同时检测.

  4. Determination of aflatoxins B1, B2, G1, and G2 in olive oil, peanut oil, and sesame oil using immunoaffinity column cleanup, postcolumn derivatization, and liquid chromatography/fluorescence detection: collaborative study.

    Science.gov (United States)

    Bao, Lei; Liang, Chengzhu; Trucksess, Mary W; Xu, Yanli; Lv, Ning; Wu, Zhenxing; Jing, Ping; Fry, Fred S

    2012-01-01

    The accuracy, repeatability, and reproducibility characteristics of a method using immunoaffinity column (IAC) cleanup with postcolumn derivatization and LC with a fluorescence detector (FLD) for determination of aflatoxins (AFs; sum of AFs B1, B2, G1, and G2) in olive oil, peanut oil, and sesame oil have been established in a collaborative study involving 15 laboratories from six countries. Blind duplicate samples of blank, spiked at levels ranging from 0.25 to 20.0 microg/kg for AF, were analyzed. A naturally contaminated peanut oil sample was also included. Test samples were extracted with methanol-water (55 + 45, v/v). After shaking and centrifuging, the lower layer was filtered, diluted with water, and filtered through glass microfiber filter paper. The filtrate was then passed through an IAC, and the toxins were eluted with methanol. The toxins were then subjected to RPLC-FLD analysis after postcolumn derivatization. Average recoveries of AFs from olive oil, peanut oil, and sesame oil ranged from 84 to 92% (at spiking levels ranging from 2.0 to 20.0 microg/kg); of AFB1 from 86 to 93% (at spiking levels ranging from 1.0 to 10.0 microg/kg); of AFB2 from 89 to 95% (at spiking levels ranging from 0.25 to 2.5 microg/kg); of AFG1 from 85 to 97% (at spiking levels ranging from 0.5 to 5.0 microg/kg); and of AFG2 from 76 to 85% (at spiking levels ranging from 0.25 to 2.5 microg/kg). RSDs for within-laboratory repeatability (RSD(r)) ranged from 3.4 to 10.2% for AF, from 3.5 to 10.9% for AFB1, from 3.2 to 9.5% for AFB2, from 6.5 to 14.9% for AFG1, and from 4.8 to 14.2% for AFG2. RSDs for between-laboratory reproducibility (RSDR) ranged from 6.1 to 14.5% for AF, from 7.5 to 15.4% for AFB1, from 7.1 to 14.6% for AFB2, from 10.8 to 18.1% for AFG1, and from 7.6 to 23.7% for AFG2. Horwitz ratio values were < or = 2 for the analytes in the three matrixes. PMID:23451385

  5. Aflatoxins and ochratoxin A in maize of Punjab, Pakistan.

    Science.gov (United States)

    Iram, Wajiha; Anjum, Tehmina; Abbas, Mateen; Khan, Abdul Muqeet

    2014-01-01

    Aflatoxin and ochratoxin levels were determined in maize samples collected from store houses of 15 districts belonging to three agro-ecological zones of Punjab, Pakistan. Toxins were extracted by Aflaochra immunoaffinity columns and analysed by high-performance liquid chromatography (HPLC). Mean moisture content of maize kernels was recorded above the safe storage level of 15%. Results indicated that aflatoxin B1 and B2 contamination was found in 97.3% and 78.9% of the collected samples, respectively. Aflatoxin G1, aflatoxin G2 and ochratoxin A were not detected in any sample. Among positive samples, 77.3% contained aflatoxin B1 and 28% aflatoxin B2, exceeding the legal limits as set by the European Union (EU) and the United States Food and Drug Administration (USFDA). It was concluded that a significant number of samples contained aflatoxin B1 and B2 above the legal limits.

  6. Development of methods for determining aflatoxins in biological material

    OpenAIRE

    Kussak, Anders

    1995-01-01

    In this thesis, it is shown how aflatoxins can be determined in biological material. The thesis is a summary of five papers. Aflatoxins are carcinogenic mycotoxins produced by Aspergillus moulds. Methods were developed for the determination of aflatoxins in samples of airborne dust and human urine collected at feed factories. For the dust samples from such agricultural products as copra, cotton seed and maize, methods were developed for the determination of aflatoxins B1, B2, G1 and G2. For u...

  7. Calcium montmorillonite clay reduces AFB1 and FB1 biomarkers in rats exposed to single and co-exposures of aflatoxin and fumonisin

    Science.gov (United States)

    Mitchell, Nicole J.; Xue, Kathy S.; Lin, Shuhan; Marroquin-Cardona, Alicia; Brown, Kristal A.; Elmore, Sarah E.; Tang, Lili; Romoser, Amelia; Gelderblom, Wentzel C. A.; Wang, Jia-Sheng; Phillips, Timothy D.

    2014-01-01

    Aflatoxins (AFs) and fumonisins (FBs) can co-contaminate foodstuffs and have been associated with hepatocellular and esophageal carcinomas in humans at high risk for exposure. One strategy to reduce exposure (and toxicity) from contaminated foodstuffs is the dietary inclusion of a montmorillonite clay (UPSN) that binds AFs and FBs in the GI tract. In this study, the binding capacity of UPSN was evaluated for AFB1, FB1 and a combination thereof in Fischer-344 rats. Rats were pre-treated with different dietary levels of UPSN (0.25 or 2%) for 1 week. Rats were gavaged with a single dose of either 0.125 mg AFB1 or 25 mg FB1/kg b.w. and a combination thereof in the presence and absence of an aqueous solution of UPSN. The kinetics of mycotoxin excretion were monitored by analyzing serum AFB1-albumin, urinary AF (AFM1), and FB1 biomarkers over a period of 72 hr. UPSN decreased AFM1 excretion by 88-97%, indicating highly effective binding. FB1 excretion was reduced, to a lesser extent, ranging between 45 to 85%. When in combination, both AFB1 and FB1 binding occurred, but capacity was decreased by almost half. In the absence of UPSN, the combined AFB1 and FB1 treatment decreased the urinary biomarkers by 67 and 45% respectively, but increased levels of AFB1-albumin, presumably by modulating its cytochrome metabolism. UPSN significantly reduced bioavailability of both AFB1 and FB1 when in combination; suggesting that it can be utilized to reduce levels below their respective thresholds for affecting adverse biological effects. PMID:24193864

  8. 免疫亲和柱萃取-液相色谱柱后衍生荧光法检测黄曲霉毒素B1、B2、G1、G2%Determination of aflatoxin B1, B2, G1 and G2 based on immunoaffinity column extraction and liquid chromatography with postcolumn derivatization and fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    郝尚东; 欧战功

    2013-01-01

    Objective:This study presents a method for simultaneous determination of aflatoxin B1,B2,G~,and G2.Methods:The samples were purified and enriched with immunoaffinity column,with the mobile phase consisting of water,methanol and acetonitrile (6∶3∶2),350 μl 4 mol/L nitric acid and 0.120 g potassium bromide,determined by fluorescent method after high performance liquid chromatography with postcolumn derivation.Results:The method is highly selective with good.linearity (r > 0.999),the linear range of the method was 0.07 μg/kg ~9.08 μg/kg for AFB1,0.02 μg/kg ~ 2.70 μg/kg for AFB2,0.07 μg/kg ~ 9.0 μg/kg for AFG1,0.03 μg/kg ~2.79 μg/kg for AFG2,the detection limit and limit of quantification values (μg/kg) were:aflatoxin B1,0.02,0.07 ; aflatoxinB2,0.01,0.02 ; aflatoxin G1,0.02,0.07 ; and aflatoxin G2,0.01,0.03,the recoveries of the samples ranged from 80% to 105%.Condusion:The results show that the method is rapid,sensitive and accurate,and it can determine AFB1,AFB2,AFG1,AFG2 simultaneously.%目的:本文研究了同时测定黄曲霉毒素B1、B2、G1和G2(AFB1、AFB2 、AFG1、AFG2)的方法.方法:使用免疫亲和柱富集净化,以每升流动相含有0.120 g溴化钾和350μ4 mol/L硝酸的水:甲醇:乙腈(6:3:2)为洗脱液,高效液相色谱电化学柱后衍生,荧光法测定.结果:方法的选择性较好,线性关系(r>0.999),线性范围:AFB1为0.07 μg/kg ~ 9.08 μg/kg; AFB2为0.02 μg/kg~2.70 μg/kg;AFG1为0.07 μg/kg~9.0 μg/kg; AFG2为0.03 μg/kg~2.79 μg/kg,相对标准偏差0.5% ~ 3.5%之间,检测限和定量限值(μg/kg):AFB1为0.02,0.07;AFB2为,0.01,0.02;AFG1为0.02,0.07;AFG2为0.01,0.03,回收率为80%~105%.结论:该方法快速、准确,能同时测定AFB1、AFB2 、AFG1、AFG2.

  9. Simultaneous Measurement of Aflatoxin B1 and Zearalenone in Gastrointestinal Chyme of Pigs and Assessment of Binding Efficacy of Mycotoxin Adsorbents%猪胃肠道食糜中黄曲霉毒素 B1和玉米赤霉烯酮检测方法的改进及其在霉菌毒素吸附剂吸附效果评价中的应用

    Institute of Scientific and Technical Information of China (English)

    安亚南; 王丹阳; 丁立人; 杨新岗; 邓亚军; 刘强

    2015-01-01

    This experiment was conducted to develop a high-performance liquid chromatographic ( HPLC ) method for quantitative determination of aflatoxin B1 and zearalenone in supernatant liquor of gastric and intesti-nal of pigs, and the method was used to evaluate binding efficacy of aflatoxin B1 and zearalenone by mycotoxin absorbents in different media. In order to eliminate the influence of complex composition in the supernatant, the experiment optimized extraction and purification of the supernatant, detection wavelengths and mobile phases of liquid chromatography. Binding efficacy of 4 mycotoxin adsorbents in a fixed mycotoxin concentration was e-valuated respectively in purified water,the supernatant liquor of gastric and intestinal chyme of pigs. The results showed as follows:1) the supernatant liquor samples were extracted with n-hexane and dichloromethane, the optimal chromatographic condition was that the fluorescent emitted light and excitation light wavelength were set as 360, 440 nm within 0 to 9 minutes and 235, 418 nm within 9 to 14 minutes, respectively, and the mo-bile phase was the mixture of acetonitrile and water with acetonitrile percentage in the range of 40% to 60%. The analytical method was quantitative determination for aflatoxin B1 from 1. 00 to 10 000. 00 ng/mL and zearalenone from 20.00 to 5 000.00 ng/mL, respectively. The linear correlation coefficient of aflatoxin B1 was 1; the limit of detection was 0.16 ng/mL, the intra-day and inter-day precision were 2.37% and 2.74%, with recoveries ranging from 89. 1% to 100. 3%. The linear correlation coefficient of zearalenone was 0. 999; the limit of detection was 2.00 ng/mL, the intra-day and inter-day precision were 4.50% and 8.84%, with recov-ery ranging from 98. 7% to 104. 1%. 2) The adsorption rates of aflatoxin B1 on P3, M3, Y1 and Y2 were 72.10%, 84.18%, 83.17% and 66.01% from gastric supernatant of pigs, and 78.15%, 88.08%, 88.12% and 67.34% from intestinal supernatant,respectively. The binding

  10. Simultaneous Detection of Aflatoxin B1、B2、G1、G2 in Fried Foods By Ultra Performance Liquid Chromatography%超高效液相色谱法同时测定油炸食品中4种黄曲霉毒素

    Institute of Scientific and Technical Information of China (English)

    张英; 蔡志斌; 郑志伟

    2011-01-01

    Objective To establish a rapid, sensitive and simple method for the determination of aflatoxin B1 、B2 、G1、G2 in fried foods by ultra performance liquid chromatography (UPLC). Methods Sample was extracted with 84% acetonitrile and cleaned- up with the MFC. The purified solution was concentrated and evaporated to dryness. Aflatoxins were derivatized with TFA and detected and quantitated by UPLC with fluorescence detection. Results The four afltoxins could be separated in 5 minutes. The linear range of the method is 0.10~ 100. 00μg/L for aflatoxin B1 ,G1, and 0.05~50. 00μg/L for aflatox B2, G2, and the lowest detection limit was 0. 04μg/kg for aflatoxin B1 、G1 and 0.02μg/kg for B2, G2. The method has been successfully applied to determination of fried foods. The recoveries of aflatoxins was in the range of 82.0% ~ 101. 2%and the relative standard deviation was < 5 %,The correlation coefficient was ≥0.999. Conclusion This method is simpie,high sensitive, accurate and rapid for aflatoxin B1 、B2 、G1 、G2 detection in fried foods.%目的 建立一种快速、灵敏、简便的超高效液相色谱法(UPLC)同时测定油炸食品中黄曲霉毒素B1(AFB1),B2 (AFB2),G1 (AFG1),G2 (AFG2)水平的方法.方法 样品用乙腈水(84:16)提取液提取后过滤,滤液经黄曲霉毒素多功能净化柱(MPC)净化,纯化液经浓缩挥干后,加三氟乙酸(TFA)衍生后,用带有荧光检测器的超高效液相色谱仪测定.结果 4种黄曲霉毒素5 min完全分离,线性范围质量浓度分别为0.10~100.00μg/L(AFB1,AFG1),0.05-50.00μg/L(AFB2,AFG2),最低检出质量分数分别为0.04μg/kg(AFB1,AFG1),0.02μg/kg(AFB2,AFG2),油炸食品回收率为82.0%~101.2%,相对标准偏差<5%,r≥0.999.结论 该方法简单、快速、灵敏度高,适用于油炸食品中4种黄曲霉毒素水平的同时测定.

  11. The effects fo complex enzy mes for detoxicated aflatoxin B1 on production performance and egg quality of laying hens%复合酶制剂降解黄曲霉毒素 B1对蛋鸡生产性能和蛋品质的影响

    Institute of Scientific and Technical Information of China (English)

    胡常英; 胡科峰; 秦慧娟; 范星; 王云鹏

    2015-01-01

    In this study, we investigated effects of aflatoxin B1 degradation complex enzymes(GOD and CAT) on production perform-ance and egg quality of laying hens fed with naturally moldy corn with AFB 1 .Two hundred and ten 180-day-old Jing Fen laying hens were randomly divided into seven groups (CK, BA1, BA2, BA3, BD1, BD2 and BD3) .The seven dietary treatments consisted of con-ventional feed without moldy corn or complex enzymes as a control diet (CK), AFB 1 content of moldy corn were 17.22,53.27 and 134.56 g/kg (BA1, BA2 and BA3), respectively and moldy corns were added into 0.5%complex enzymes(BD1, BD2 and BD3). Results showed the production performance and egg quality of laying hens in BA 1,BA2 and BA3 groups were all decreased in different degrees, the production performance and egg quality of laying hens in BD 1,BD2 groups at three phases(T1,T2 and T3) and BD3 at the first phase(T1) of experimental time continuously were all increased compared to that in CK groups .In BD3 group was compared with in BA3 group at third phase(T3) of experimental time continuously , and the monthly output of eggs has gone up from 9 to 17, the hatching rate of fertilized eggs has gone up from 41%to 57%, the yolk weight has gone up from 14.34 g to15.53 g , the egg weight has gone up from 42.39 g to 48.49 g, the eggshell thickness has gone up from 0.28mm to 0.31mm.As a result , the complex enzymes could reduce or almost eliminate the negative effects of AFB 1 on production performance and egg quality of laying hens .%试验旨在研究葡萄糖氧化酶(Glucose Oxidase, GOD )和过氧化氢酶(catalase, CAT)复合酶制剂降解污染饲料中黄曲霉毒素B1Aflatoxin B1, AFB1)对蛋鸡生产性能和蛋品质的影响。选用210只180日龄京粉蛋鸡,随机分为7组( CK、BA1、BA2、BA3、BD1、BD2和BD3),7个处理组分别是饲喂不添加污染玉米和复合酶的CK组,污染饲料中含有17.22,53.27和134.56μg/kg AFB1的BA1,BA2和BA3

  12. Aflatoxin M1 contamination of human breast milk in Isfahan, Iran

    Science.gov (United States)

    Jafarian-Dehkordi, Abbas; Pourradi, Nasibeh

    2013-01-01

    Background: During the last decades there has been great attention paid to aflatoxins. They are highly toxic, immunosuppressive, mutagenic, teratogenic, and carcinogenic compounds. Aflatoxin M1 (AFM1), a hydroxylated metabolite of aflatoxin B1 (AFB1), is formed in the liver and excreted into the breast milk. It is considered to cause certain hygienic risks for infant health. The aim of this study was to evaluate the presence of the AFM1 in the breast milk using AFM1 in milk as a biomarker for exposure to aflatoxin B1 and determine the level of AFM1 contamination in the lactating mothers in Isfahan, Iran. Materials and Methods: This study was carried out on 80 lactating women randomly selected from two urban health centers. Mother's milk samples and information on food intake were collected from the participants using structured food-frequency questionnaire. Breast milk samples were tested for AFM1 by a competitive ELISA technique. Results: Our findings showed that only one sample was contaminated with AFM1 with concentrations of 6.8 ng/L. However, the AFM1 level in this sample was lower than the maximum tolerable limit (25 ng/L) accepted by the European Communities and Codex Alimentarius. Conclusion: Although the concentration of AFM1 in none of the samples was higher than the acceptable level, the presence of AFM1 in only one of them confirms the need for developing strategies to reduce exposure to aflatoxin in foods and to carry out biological monitoring of aflatoxins as a food quality control measure routinely. PMID:24524032

  13. Determination of aflatoxin B1, B2, G1, G2 and M1 in corn by high performance liquid chromatographytandem mass spectrometry%固相萃取-高效液相色谱/串联质谱检测谷物中黄曲霉毒素B1、B2、G1、G2和M1

    Institute of Scientific and Technical Information of China (English)

    王岩松; 范世华; 李华; 张凤清; 贺明睿; 崔相勇

    2011-01-01

    建立了高效液相色谱/串联质谱(LC-MS/MS)同时检测谷物中的黄曲霉毒素B1、B2、G1、G2和M1的方法.并优化了液相色谱条件和质谱的相关参数.谷物样品经研磨成粉末后,直接经甲醇-水( V∶V=10∶90)提取,Oasis HLB固相萃取净化,乙腈-水(0.2%甲酸)梯度洗脱,选择电喷雾离子源(ESI),正离子扫描多反应监测( MRM)模式,外标法定量.黄曲霉毒素B1、B2、G1、G2和M1的定量下限分别为0.1、0.1、0.2、0.3、0.2 ng/g,平均回收率在74.6%~89.6%之间,测试精密度(RSD)在5.2%~11.3%之间.方法已用于谷物样品检测,黄曲霉毒素残留量最高达到B1 5.86 ng/g、G1 8.6 ng/g、M1 4.0 ng/g,B2和G2未检出.%A sensitive method was developed for the determination of aflatoxin B,, B2, G,, G2 and M, in corn by high performance liquid chromatography-tandem mass spectrometry ( HPLC-MS/MS). Analysis conditions of liquid chromatography and mass spectrometry were optimized. Aflatoxins were extracted with 10% ( V/V) methanol-water after corn was grinded into powder, purified through an Oasis HLB cartridge, eluted by methanol and water containing 0. 2% formic acid, and determinated by electrospray ionization in positive ion mode and using multiple reaction monitoring (MRM). The limits of quantification for aflatoxin B,, B2, G1, G2 and M1 were 0. 1 ng/g, 0. 1 ng/g, 0.2 ng/g, 0.3 ng/g and 0.2 ng/g, the correlation coefficients were more than 0.9975 in respective linear ranges, and recoveries for analytes in different matrixs were 74. 6% ~ 89. 6% and the relative standard deviations (RSD) in different concentration levels were all less than 11. 3%. The proposed method was sensitive to meet the requirements for monitoring aflatoxin B1, B2, G1, G2 and Ml in corn.

  14. Incidence and Level of Aflatoxins Contamination in Medicinal Plants in Korea

    OpenAIRE

    Lee, Sung Deuk; Yu, In Sil; Jung, Kweon; Kim, Yeon Sun

    2014-01-01

    During 2011~2013, a total of 729 samples for 19 types of medicinal plant were collected from Seoulyekryungsi in Seoul, Korea, and investigated for the presence of aflatoxins. The samples were analyzed using immunoaffinity column cleanup and high-performance liquid chromatography coupled to a fluorescence detector after post-column derivatization. Aflatoxins were found in 124 out of the 729 analyzed samples: 65 containing aflatoxin B1 (AFB1), 24 with aflatoxin B2 (AFB2), 15 with aflatoxin G1 (...

  15. The occurrence of aflatoxins in selected spices and dried fruits

    Directory of Open Access Journals (Sweden)

    Renata Stanisławczyk

    2013-03-01

    Full Text Available Mycotoxins are toxic substances formed by fungi which are a potential danger for human and animal health. The aim of this study was to determine the contamination with the aflatoxin B1 and aflatoxins sum B1, B2, G1, G2 in selected spices and dried fruits available in trade in the Podkarpackie province. The studies confirm the widespread occurrence of aflatoxin B1 and aflatoxins sum B1, B2, G1, G2 in spices, spices mixtures and dried fruits, however, in none of the tested samples there was exceeded the permissible concentration of aflatoxin B1 and aflatoxins sum B1, B2, G1, G2. The highest content of aflatoxin B1 was determined in red pepper and universal spices mixture while the smallest in figs and raisins. The monitoring of aflatoxins sum B1, B2, G1, G2  showed similar results. The highest level was determined in universal spices mixture and in red pepper.

  16. Effect of gamma radiation on the inactivation of aflatoxin B1 in food and feed crops Efeito da radiação gama na inativação de aflatoxina B1 em alimentos e ração

    Directory of Open Access Journals (Sweden)

    I. Ghanem

    2008-12-01

    Full Text Available Samples of food crops (peanut, peeled pistachio, unpeeled pistachio, rice, and corn and feed (barley, bran, corn were autoclave-sterilized, and inoculated with 10(6 of spore suspension of an isolate of Aspergillus flavus fungus known to produce aflatoxin B1 (AFB1 . Following a 10-day period of incubation at 27 C to allow for fungal growth, food and feed samples were irradiated with gamma radiation at the doses 4, 6, and 10 kGy. Results indicated that degradation of AFB1 was positively correlated with the increase in the applied dose of gamma ray for each tested sample. At a dose of 10 kGy percentages of AFB1 degradation reached highest values at 58.6, 68.8, 84.6, 81.1 and 87.8% for peanuts, peeled pistachios, unpeeled pistachios, corn and rice samples, respectively. In feed samples percentages of AFB1 degradation were 45, 66, and 90% in barley, 47, 75, and 86% in bran, and 31, 72, and 84% in corn for the doses of 4, 6, and 10 kGy, respectively. AFB1 degradation in food samples correlated negatively with oil content in irradiated samples. Thus, in peanuts, which contained the highest oil content, percentage of AFB1 degradation at 10 kGy was not more than 56.6%, whereas, the corresponding value in corn, which contained the lowest oil content, reached as high as 80%. The above results indicate the possibility of using gamma radiation as a means of degradation of AFB1 in food and feed crops to levels lower than the maximum allowed levels.Amostras de alimentos (amendoim, pistache descascada, pistache com casca, arroz e milho e de ração (cevada, farelo de trigo e milho foram esterilizadas por autoclavação e inoculadas com uma suspensão de esporos (10(6 de um isolado de Aspergillus flavus produtor de aflatoxina B1 (AFB1. Após incubação por 10 dias a 27ºC para multiplicação do fungo, as amostras foram irradiadas com radiação gama nas doses de 4, 6 e 10 kGy. Os resultados indicaram que a degradação da AFB1 correlacionou-se positivamente

  17. Preliminary study on the DNA damage of human embryo liver cell in vitro induced by aflatoxin B1%黄曲霉毒素B1致体外人胚肝细胞DNA损伤的初步研究

    Institute of Scientific and Technical Information of China (English)

    方宁烨; 刘丹丹; 肖德强; 陈敏玫; 王志清; 曾高峰; 鲁力

    2012-01-01

      目的研究黄曲霉毒素 B1(AFB1)对人胚肝细胞(L-02细胞)的毒性作用特点,为建立 AFB1致L-02细胞 DNA 损伤的体外实验模型打下基础.方法以 L-02细胞为靶细胞,采用完全随机实验设计将其分为3组:(1)空白对照组;(2)溶剂对照组;(3)AFB1染毒组,分别使用5,10,20,40mg/L 4个剂量染毒.各组细胞染毒培养24h 后,收集细胞进行单细胞凝胶电泳(SCGE)试验,观察细胞 DNA 损伤情况;同时收集细胞培养上清液,用全自动生化分析仪测定谷草转氨酶(AST)和谷丙转氨酶(ALT)含量.结果(1)SCGE 试验结果显示,染毒浓度达到40mg/L 时 L-02细胞出现 DNA 损伤,其尾长和 Olive 尾矩值与空白对照和溶剂对照相比有显著差异(P 0.05).结论 AFB1染毒浓度达到40mg/L 的浓度时,就能够诱导 L-02细胞 DNA 出现损伤,但肝功能 AST 和 ALT 却未见异常.实验结果表明,在 L-02细胞未出现急性损伤的低浓度 AFB1染毒状态下,其致肝细胞 DNA 损伤的作用就已显现.%  Objective To study the toxic effect of aflatoxin B1 on the human embryo liver cell(L-02 cell) in order to set up an in vitro model of L-02 cell DNA damage induced by AFB1. Methods L-02 cell was choosed as a target cell, and using a completely randomized design, the cell was divided into three groups:(Ⅰ)blank control group;(Ⅱ)solvent control group;(Ⅲ)AFB1 exposed groups, with 5, 10, 20, 40mg/L four dosage contaminations. After each group of cells was cultured for 24 hours, SCGE was performed on the collected cell,and the situation of cell damage was also observed . At the same time, the cell culture supernatant of each group was collected ,the values of AST and ALT were determined by the automatic biochemistry analyzer. Results (Ⅰ) SCGE showed that only 40mg/L exposure group occurred a cell DNA damage, Taillength and Olive tailmoment in this group( P 0.05) compared with blank control group. Conclusion SCGE has shown that 40mg/L AFB1 is able

  18. Contamination of aflatoxin B1 in peanuts around harvest from counties of Linyi in Shandong province%山东临沂部分县区花生收获前后黄曲霉毒素B1污染情况研究

    Institute of Scientific and Technical Information of China (English)

    苏祥; 刘建伟; 韩小敏; 秦泽明; 温红玲; 宋艳艳; 李凤琴; 赵丽

    2015-01-01

    目的:了解花生从收获前到收获、储存期间黄曲霉毒素B1(AFB1)污染的环节,为预防花生AFB1的污染提供科学依据。方法2014年,在山东临沂市选择花生种植范围较广的5个县,分别于收获前1~2周、收获时、收获晾干后1个月和储存3个月采集花生样本共260份,酶联免疫法(ELISA)检测花生 AFB1的污染情况,并对结果进行分析。结果260份花生样品中,收获前、收获时样品中AFB1均小于2μg/kg,收获晾干后1个月和储存3个月各有1份样品AFB1含量严重超标,分别是81.07μg/kg和为254.27μg/kg,阳性率均为1.27%;总阳性率为0.77%。结论2014年临沂部分地区花生中AFB1阳性率较低,污染主要发生在收获后的晾晒和储存期,提示科学晾晒和储存是预防花生AFB1污染的重要措施之一。%Objective To investigate aflatoxin B1 (AFB1) contamination of peanuts from pre-harvest, harvest period to storage period, and to provide a scientific basis to prevent contamination of AFB1 in peanuts. Methods In 2014, five counties of Linyi in Shandong province were chosen, and 260 peanut samples were collected in 1~2 weeks before harvest, harvest, post-harvest 1 month and 3 months storage, standardized enzyme-linked immunosorbent assay (ELISA) kits were used to detect AFB1 content in peanuts, then, the results were analyzed. Results In 260 peanut samples, AFB1 concentrations from all pre-harvest and harvest peanuts were less than 2μg/kg, AFB1 of 2 samples from 1 month after drying and 3 months storage were out of limits, the values were 81.07 μg/kg and 254.27 μg/kg, respectively. The positive rate was 1.27%, and total positive rate was 0.77%. Conclusion AFB1 contamination of peanuts in this region occurred mostly in drying and storage period after harvest, scientific drying and storage were one of the most important measures to prevent AFB1 contamination in peanuts.

  19. Comparison of methods by TLC and HPTLC for determination of aflatoxin M1 in milk and B1 in eggs Comparação de metodologia para análise de aflatoxina M1 em leite e aflatoxina B1 em ovos por CCD e CCDAE

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    V. M. Scussel

    2003-12-01

    Full Text Available Milk and egg matrixes were assayed for aflatoxin M1 (AFM1 and B1 (AFB1 respectively, by AOAC official and modified methods with detection and quantification by thin layer chromatography (TLC and high performance thin layer chromatography (HPTLC. The modified methods: Blanc followed by Romer, showed to be most appropriate for AFM1 analysis in milk. Both methods reduced emulsion formation, produced cleaner extracts, no streaking spots, precision and accuracy improved, especially when quantification was performed by HPTLC. The use of ternary mixture in the Blanc Method was advantageous as the solvent could extract AFM1 directly from the first stage (extraction, leaving other compounds in the binary mixture layer, avoiding emulsion formation, thus reducing toxin loss. The relative standard deviation (RSD% values were low, 16 and 7% when TLC and HPTLC were used, with a mean recovery of 94 and 97%, respectively. As far as egg matrix and final extract are concerned, both methods evaluated for AFB1 need further studies. Although that matrix leads to emulsion with consequent loss of toxin, the Romer modified presented a reasonable clean extract (mean recovery of 92 and 96% for TLC and HPTLC, respectively. Most of the methods studied did not performed as expected mainly due to the matrixes high content of triglicerides (rich on saturated fatty acids, cholesterol, carotene and proteins. Although nowadays most methodology for AFM1 is based on HPLC, TLC determination (Blanc and Romer modified for AFM1 and AFB1 is particularly recommended to those, inexperienced in food and feed mycotoxins analysis and especially who cannot afford to purchase sophisticated (HPLC,HPTLC instrumentation.Aflatoxinas M1 (AFM1 e B1 (AFB1 foram analisadas em leite e ovos respectivamente, por diferentes métodos oficiais da AOAC e modificações usando detecção por cromatografia em camada delgada (CCD e CCD de alta eficiência (CCDAE. Os métodos modificados: Blanc e Romer

  20. β-环糊精及其衍生物对黄曲霉毒素B_1荧光增强机理研究%Mechanisms Underlying Fluorescence Enhancement of Aflatoxin B_1 by β-Cyclodextrin and Its Derivatives

    Institute of Scientific and Technical Information of China (English)

    张敏; 郭婷; 刘馨; 肖洁; 张宇昊; 马良

    2012-01-01

    The mechanisms underlying fluorescence enhancement of aflatoxin B1(AFB1) by β-cyclodextrin(β-CD) and its derivatives were explored using spectroscopy,Benesi-Hidebrand analysis and thermodynamics.Moreover the effects of methanol volume percentage,M-β-CD concentration,reaction time and interfering ions on fluorescence enhancement were analyzed.β-CD and its six derivatives showed an inclusion ratio to AFB1 of 1:1 at low concentrations and 2:1 at high concentrations.The thermodynamic parameters entropy change(ΔS),enthalpy change(ΔH) and free energy change(ΔG) for maximum inclusion constant between M-β-CD and AFB1 were negative,suggesting that the inclusion reaction is exothermic and can occur spontaneously.Enthalpy change was the dominant force during the formation of inclusion complexes.UV absorption spectra and KI quenching experiments showed that AFB1 could enter M-β-CD cavity to protect fluorescence.Therefore,β cyclodextrin and its derivatives can allow considerable enhancement in the fluorescence emission intensity of AFB1 by the formation of supramolecular inclusion complexes,resulting in increased sensitivity of AFB1fluorescence analysis.%利用光谱法、Benesi-Hildebrand法、热力学方法研究β-环糊精(β-CD)及其衍生物对黄曲霉毒素B1(AFB1)荧光增强机理,探讨溶剂中甲醇体积比、M-β-CD浓度、时间、干扰离子等因素对荧光增强作用的影响。根据Benesi-Hildebrand法确定7种β-环糊精及其衍生物在低浓度时与AFB1包络比为1:1,高浓度时包络比为2:1。采用热力学方法计算了包合常数最大的M-β-CD与AFB1包合过程的熵变(ΔS)、焓变(ΔH)及自由能变化(ΔG)均为负值,说明包合反应是放热反应且能自发进行,焓变是形成超分子包络物的主要驱动力。紫外吸收光谱及KI淬灭实验表明AFB1进入M-β-CD空腔从而使荧光得到保护。得出结论:7种β-环糊精及其衍生物与AFB1通过形成超分子

  1. EFFECT OF GINKGO BILOBA LEAF EXTRACT ON HEPATOCARCINOGENESIS INDUCED BY AFLATOXIN B1 IN WISTAR RATS%银杏叶提取物对黄曲霉毒素B1诱发Wistar大鼠肝癌的影响

    Institute of Scientific and Technical Information of China (English)

    蒿艳蓉; 杨芳; 曹骥; 欧超; 李媛; 杨春; 段小娴; 苏建家

    2010-01-01

    [目的]通过建立黄凿霉毒素B1(Aflatoxin B1,AFB1)诱发Wistar大鼠肝细胞癌(HCC)模型,用银杏叶提取物(extract of Ginkgo Biloba Leaf,EGb761)进行干预,观察EGb761对AFB1致大鼠HCC作用的影响. [方法]实验大鼠按体重随机分成3组:A组(AFB1组)、B组(AFBI+EGb761组)和C组(空白对照组).于实验第14、28、42及55周给予肝活检并于第64周全部处死大鼠,观察大鼠肝脏γ-GY灶及HCC发生情况. [结果]在出癌前期,第42及第55周时,A组每个γ-GT阳性灶的面积分别为(7.95±0.30)mm2及(17.87±0.71)mm2,单位面积(cm2)灶的个数分别为0.35±0.006及0.26±0.004,灶总面积分别为(2.54±0.05)mm2/cm2及(4.68±0.12)mm2/cm2;B组在相同时段的相应指标,分别为(4.65±0.16)mm3及(9.03 4±0.35)mm2, (0.21 4±0.006)及(0.20±0.005),(0.97±0.03)mm2/cm2及(1.62±0.06)mm2/cm2.除了55周A、B两组的单位面积γ-GY阳性灶的个数无统计学差异外,各时段B组各项指标都显著小于A组(P=0.000).两个实验组共有28只大鼠发生恶性肿瘤,对照组无肿瘤发生.B组HCC诱发率(7/26,26.92%)明显低于A组(19/25,76%) (P=0.000).另外,HCC及其他肿瘤的发生时间,B组明显推迟.[结论]EGb761能明显抑制AFB1诱发大鼠肝的癌前病变及延迟癌前病变向癌发展.

  2. Aflatoxins as a cause of hepatocellular carcinoma.

    Science.gov (United States)

    Kew, Michael C

    2013-09-01

    Aflatoxins, metabolites of the fungi Aspergillus flavus and Aspergillus parasiticus, are frequent contaminants of a number of staple foods, particularly maize and ground nuts, in subsistence farming communities in tropical and sub-tropical climates in sub-Saharan Africa, Eastern Asia and parts of South America. Contamination of foods occurs during growth and as a result of storage in deficient or inappropriate facilities. These toxins pose serious public health hazards, including the causation of hepatocellular carcinoma by aflatoxin B1. Exposure begins in utero and is life-long. The innocuous parent molecule of the fungus is converted by members of the cytochrome p450 family into mutagenic and carcinogenic intermediates. Aflatoxin-B1 is converted into aflatoxin B1-8,9 exo-epoxide, which is in turn converted into 8,9-dihydroxy-8-(N7) guanyl-9-hydroxy aflatoxin B1 adduct. This adduct is metabolized into aflatoxin B1 formaminopyrimidine adduct. These adducts are mutagenic and carcinogenic. In addition, an arginine to serine mutation at codon 249 of the p53 tumor suppressor gene is produced, abrogating the function of the tumor suppressor gene, and contributing to hepatocarcinogenesis. Aflatoxin B1 acts synergistically with hepatitis B virus in causing hepatocellular carcinoma. A number of interactions between the two carcinogens may be responsible for this action, including integration of hepatitis B virus x gene and its consequences, as well as interference with nucleotide excision repair, activation of p21waf1/cip1, generation of DNA mutations, and altered methylation of genes. But much remains to be learnt about the precise pathogenetic mechanisms responsible for aflatoxin B1-induced hepatocellular carcinoma as well as the interaction between the toxin and hepatitis B virus in causing the tumor.

  3. Aflatoxins and heavy metals in animal feed in Iran.

    Science.gov (United States)

    Eskandari, M H; Pakfetrat, S

    2014-01-01

    The occurrence of aflatoxin (aflatoxin B1, aflatoxin B2, aflatoxin G1 (AFG1) and aflatoxin G2 (AFG2)) and heavy metal (Pb, Cd, As and Hg) contamination was determined in 40 industrially produced animal feed samples which were collected from the southwest of Iran. The results indicated that 75% of samples were contaminated by four aflatoxins and the level of AFB1 and sum of aflatoxins were higher than the permissible maximum levels in Iran (5 and 20 µg kg(-1), respectively) in all feed samples. A positive correlation was found between four types of aflatoxins in all the tested samples (p < 0.01) and the positive correlation between AFG1 and AFG2 was significant (r(2) = 0.708). All feed samples had lead concentrations lower than the maximum EU limit, while 5%, 17% and 42.5% of feed samples had As, Cd and Hg concentrations higher than the maximum limits, respectively.

  4. Aflatoxins in foods

    Directory of Open Access Journals (Sweden)

    Amedeo Pietri

    2007-03-01

    Full Text Available Aflatoxins are mycotoxins produced by Aspergillus flavus and A. parasiticus. The aflatoxin group is comprised of aflatoxin B1 (AFB1, B2, G1 and G2. In addition, aflatoxin M1 (AFM1, a hydroxylated metabolite of AFB1, is excreted in the milk of dairy cows consuming an AFB1-contaminated ration. AFB1 has shown extreme acute and chronic toxicity and carcinogenic activity in animals; the acute toxicity of AFM1 is nearly equal to that of AFB1, but its potential carcinogenic hazard is about one order of magnitude less than that of AFB1. The International Agency for Research on Cancer classified AFB1 as a human carcinogen (group 1 and AFM1 as a possible carcinogen (group 2A. Recently, the possibility of a synergistic carcinogenic interaction between HBV chronic infection and dietary exposure to AFB1 arose from the observation of their co-existence in countries with high incidences of HCC and was confirmed by further experimental and epidemiological studies. However, the carcinogenic potency of AFB1 is considered much lower in populations where chronic hepatitis infections are rare. For the first time in 2003, significant problems arose in Italy, due to the aflatoxin contamination of maize. The summer was extremely hot and dry and A. flavus is very competitive under these conditions as the plants are stressed. Maize grain is normally utilized in the food supply for dairy cows and as such led to the severe and widespread contamination of milk with AFM1. In the following years (2004-2006, different climatic conditions as well as better compliance with guidelines by farmers, led to a dramatic reduction of the problem.

  5. Hepatitis infections, aflatoxin and hepatocellular carcinoma

    Directory of Open Access Journals (Sweden)

    Pierre Hainaut

    2007-02-01

    with hot, humid climates, including maize, peanuts and cottonseeds. The toxins are present at significant levels in crops at the time of harvest but their concentration further increases under poor conditions of long term food storage, in particular during the rain season. Thus, in these regions, most inhabitants of rural areas are highly exposed to aflatoxins, with seasonal variations reflecting the consumption of stored versus fresh foodstuff. Population-based surveys have demonstrated the presence of serum aflatoxin-albumin adducts in over 95% of the normal population in The Gambia, West Africa. Exposure starts in the perinatal period, through in utero transfer and breast-feeding, and continues throughout life, mainly from consumption of peanuts. Time patterns of aflatoxin-albumin adduct levels correlate with the seasonal availability of peanuts.

    There is strong experimental evidence that aflatoxins are potent hepatocarcinogens in rodents. In humans, there are good ecological correlations between the risk of HCC and the presence of biomarkers of aflatoxin exposure in serum or in urine .The most significant carcinogenic aflatoxin is B1 (AFB1, which is the most abundant in the diet. AFB1 is metabolized in the liver by several CYP450 enzymes (mainly 1A2 and 3A4 to a reactive AFB1-8,9- exo-epoxide (Mace et al. 1997. This metabolite generates a primary DNA adduct (8-9, dihydro-8-(N7-guanyl-9-hydroxyaflatoxin; AFB1-N7-Gua, naturally converted to two secondary lesions, an apurinic (AP site and a stable, AFB1-formamidopyrimidine (AFB1-FAPY adduct The latter is considered as the most mutagenic lesion (Smela et al. 2002. The sequence context of codon 249 (AGGCC represents a site of intermediate affinity for the formation of AFB1-induced lesions. Other codons in TP53, including some codons that are ";hotspots"; in many cancers (codon 245, 248 and 273, have a similar or even greater affinity

  6. Development and in-house validation of a robust and sensitive solid-phase extraction liquid chromatography/tandem mass spectrometry method for the quantitative determination of aflatoxins B1, B2, G1, G2, ochratoxin A, deoxynivalenol, zearalenone, T-2 and HT-2 toxins in cereal-based foods.

    Science.gov (United States)

    Lattanzio, Veronica M T; Gatta, Stefania Della; Suman, Michele; Visconti, Angelo

    2011-07-15

    A sensitive and robust liquid chromatography/tandem mass spectrometry (LC/MS/MS) method was developed for the simultaneous determination of aflatoxins (B(1), B(2), G(1), G(2)), ochratoxin A, deoxynivalenol, zearalenone, T-2 and HT-2 toxins in cereal-based foods. Samples were extracted with a mixture of acetonitrile/water (84:16, v/v) and cleaned up through a polymeric solid-phase extraction column. Detection and quantification of the nine mycotoxins were performed by reversed-phase liquid chromatography coupled with electrospray ionization triple quadrupole mass spectrometry (LC/ESI-MS/MS), using fully (13)C-isotope-labelled mycotoxins as internal standards. The method was validated in-house for five different cereal processed products, namely barley, oat and durum wheat flours, rye- and wheat-based crisp bread. Recoveries and repeatability of the whole analytical procedure were evaluated at contamination levels encompassing the EU maximum permitted levels for each tested mycotoxin. Recoveries ranged from 89 to 108% for deoxynivalenol, from 73 to 114% for aflatoxins, from 85 to 114% for T-2 and HT-2 toxins, from 64 to 97% for zearalenone, from 74 to 102% for ochratoxin A. Relative standard deviations were less than 16% for all tested mycotoxins and matrices. Limits of detection (signal-to-noise ratio 3:1) ranged from 0.1 to 59.2 µg/kg. The trueness of the results obtained by the proposed method was demonstrated by analysis of reference materials for aflatoxins, deoxynivalenol, zearalenone. The use of inexpensive clean-up cartridges and the increasing availability of less expensive LC/MS/MS instrumentation strengthen the potential of the proposed method for its effective application for reliable routine analysis to assess compliance of tested cereal products with current regulation. PMID:21638363

  7. Effects of Ginkgo biloba extract on relative protein expressions during hepatocarcinogenesis aflatoxin B1-induced in Wistar rats%银杏叶提取物对黄曲霉毒素B1所致大鼠肝癌相关蛋白表达影响的研究

    Institute of Scientific and Technical Information of China (English)

    杨涛; 莫钦国; 欧超; 苏建家

    2011-01-01

    OBJECTIVE: To investigate the effects of Ginkgo biloba extract (extract 761 from Ginkgo giloba, EGb761) on relative protein expressions during hepatocarcinogenesis induced by aflatoxin Bl (AFB1) in rats. METHODS: The 71 rats were divided into three groups: Group AFB1 (28), Group AFBl + EGb761(29) and Group control(14). The animals were sacrificed at the end of the experiment. The incidence of liver cancers in each group was stated. The expressions of heparan sulfate proteoglycan(MXR7), Cycloxygenase-2 (COX-2 ) and Cyclin dependent kinase inhibitor (PI6) were observed by immunohistochemistry staining in above samples. RESULTS: The rate of hepatocellular carcinoma (HCC) in group AFB1 were significantly higher than group AFB1 + EGb761(76% vs 28%, X2 = 10. 602,P<0. 05). No tumor developed in the control animals. The expression of MXR7 protein in group AFB1 + EGb761 were significantly lower than group AFB1 (1. 488±1. 178) vs (5. 133±2. 725),(F=18. 638,P<0. 001). Pl6 protein in group AFB1 + EGb761 were significantly higher than group AFB1 (2. 456 ±1.014) vs (1. 533 ±0. 856), (F = 24. 091,P=0. 03). The difference of COX-2 protein expression in group AFB1 and group AFB1 + EGb761 had no significant (3. 305±0. 566) vs (2. 704 ± 0. 431) , (F= 9. 352, P = 0. 783). CONCLUSION: Ginkgo biloba extract reduces MXR7 protein expression in rats and enhance the expression of p16 protein, and it may be inhibited the occurrence of liver cancer induced by AFB1.%目的:探讨银杏叶提取物(extract 761 from Ginkgo giloba,EGb761)对黄曲霉毒素B1(aflatox B1,AFB1)致大鼠肝癌后相关蛋白表达的影响.方法:71只大鼠随机分为AFB1组(28只),AFB1+EGb761组(29只),空白对照组(14只).第64周观察大鼠肝癌发生率,应用免疫组化法检测硫酸乙酰肝素类蛋白聚糖(MXR7)、环氧化酶(COX-2)以及细胞周期蛋白依赖性蛋白激酶抑制蛋白(p16)的表达情况.结果:AFB1组肝癌发生率明显高于AFB1+EGb761组(76%vs28%),X2=10.602,P<0

  8. Reduction of toxic effects of aflatoxin B1 by using baker yeast (Saccharomyces cerevisiae in growing broiler chicks diets Redução dos efeitos tóxicos da aflatoxina B1, utilizando-se levedura de panificação (Saccharomyces cerevisiae, na dieta de pintos de corte em crescimento

    Directory of Open Access Journals (Sweden)

    Kemal Çelýk

    2003-06-01

    Full Text Available This study was carried out to investigate the effects of adding baker yeast (BY, chlortetracycline (CTC and both BY + CTC to a control diet containing 200 ng/g of aflatoxin B1 (C + AFB1 on performance, serum parameters and pathologyc alterations of broilers. A total 100 chicks (Ross PM 3 were divided into five groups in individual cages and each containing 20 animals. BY, a rich source of protein and vitamin B complex, was mixed into the diets at 2.0 %, CTC was mixed into the diet at 2.5 ng/g. Feed consumption, body weight and feed efficiency were recorded weekly. Serum parameters and pathologyc alterations were determined at the end of the study. Dead animals were recorded daily. Liver changes were clearly apparent in the C+AFB1and C+ AFB1+CTC most of the livers were enlarged, yellow and had pethecial hemorrhages. Canalicula cholestosis was absent in group C+AFB1 and C+ AFB1+CTC, but not others. When compared to the control (C group, alkaline phosphatase (ALP, appear to be significantly increased in the C+AFB1 and C+CTC+ AFB1 groups. Serum glutamic oxalacetic transaminase (GOTwas increased in C+AFB1 birds. Serum alphaphetoprotein was not affected by the treatments. Feed consumption and body weight were significantly reduced in group AFB1. Birds receiving BY + AFB1, CTC + AFB1 and BY + CTC + AFB1 had a significantly higher body weight than group C+AFB1. Feed efficiency was better in group CTC + AFB1 than the others. The findings of this research suggest tha BY (2% can partly counteract some of the toxic effects of AFB1.Este estudo foi desenvolvido para avaliar os efeitos da adição de Levedura de panificação (BY e cortetraciclina (CTC e ambos BY+CTC a uma dieta controle © contendo 200 ng/g de aflatoxina B1 (C+AFB1 sobre desempenho, parâmetros séricos e alterações patológicas de frangos de corte. Um total de 100 pintinhos (Ross PM3 foi dividido em cinco grupos, em gaiolas individuais, contendo 20 animais para cada grupo. A levedura de

  9. Inhibition of aflatoxin production by selected insecticides.

    Science.gov (United States)

    Draughon, F A; Ayres, J C

    1981-04-01

    The insecticide naled completed inhibition production of aflatoxins B1, B2, G1, and G2 by and growth of Aspergillus parasiticus at a 100-ppm (100 microgram/ml) concentration. The insecticides dichlorvos, Landrin, pyrethrum, Sevin, malathion, and Diazinon significantly (P = 0.05) inhibited production of aflatoxins at a 100-ppm concentration. However, at a concentration of 10 ppm, significant inhibition in production of aflatoxins was found only with naled, dichlorvos, Sevin, Landrin, and pyrethrum. Dichlorvos, Landrin, Sevin, and naled inhibited growth of A. parasiticus by 28.9 , 18.9, 15.7, and 100%, respectively, at 100 ppm. Stimulation of growth was observed when diazinon was added to cultures. Aflatoxin B1 was most resistant to inhibition by insecticides, followed by G1, G2, and B2, respectively. PMID:6786222

  10. Fate of Aflatoxin M1 during cheese whey processing

    OpenAIRE

    Mendonça, Carla; Venâncio, Armando

    2004-01-01

    Aflatoxins are a group of naturally occurring toxins, which are secondary metabolites of some Aspergillus spp. When lactating animals ingest aflatoxin B1 (AFB1) contaminated feedstuffs, aflatoxin M1 (AFM1) may be excreted to milk. Thus, AFM1 represents a potential hazardous to humans via consumption of milk and milk products. AFM1 is less mutagenic and carcinogenic than AFB1 but it exhibits high genotoxic activity. The maximum admissible level of this mycotoxin in raw milk, ...

  11. HPLC-柱后光化学衍生法检测花生酱中黄曲霉毒素%Determination of Aflatoxin B1,B2,G1 and G2 in Peanut Butter by HPLC with On-Line Post-Column Photochemical Derivatization

    Institute of Scientific and Technical Information of China (English)

    邵丽; 王晓; 滕振勇

    2015-01-01

    建立高效液相色谱-在线柱后光化学衍生-荧光检测器检测花生酱中黄曲霉毒素B1、B2、G1、G2的含量.样品以乙腈-水(80:20)溶液提取,经免疫亲和柱净化后,利用在线柱后光化学衍生-HPLC-FLD进行分析测定.结果:在优化条件下,黄曲霉毒素B1、G1在0.30 mg/L~10 mg/L,黄曲霉毒素B2、G2在0.06 mg/L~3.0mg/L线性关系良好,r>0.998 ,回收率80%~101%,RSD0.998. The recoveries ranged between 80 % and 101 % ,with RSDs all bellow 5.9 %. LOD of aflatoxin B1,B2,G1 and G2 were 0.10,0.03,0.15,0.04μg/kg,respectively.

  12. Fungal Biodeterioration, Aflatoxin Contamination, and Nutrient Value of "Suya Spices".

    Science.gov (United States)

    Jonathan, Segun Gbolagade; Adeniyi, Mary Adejoke; Asemoloye, Michael Dare

    2016-01-01

    This work aimed to analyze the nutrient values, examine the biodeteriorating fungi biota, and analyze the mycotoxin contents of "Suya spices." Fungi with highest percentage occurrence on all the samples are Aspergillus niger, Aspergillus flavus, Aspergillus parasiticus, Aspergillus ochraceus, Fusarium sp., Rhizopus stolonifer, yeast, and Trichoderma koningii. Nutrient composition of the samples is significantly different statistically (P Aflatoxin analysis of the samples revealed that they all contain aflatoxin in varying amount but no detectible aflatoxin content in the control. 59.54% of the detected aflatoxin is aflatoxin B1 with highest recorded in Agbowo, Mokola, and Sango samples (i.e., 28.03, 22.44, and 13.8 μg/kg, resp.). 4.78% of the aflatoxin is aflatoxin B2 which is only found in Sango and Mokola samples (3.59 and 2.6 μg/kg, resp.). 32.76% of aflatoxin is aflatoxin G1 with the highest found in Agbowo and Mokola samples (i.e., 18.63 and 10.41 μg/kg, resp.). 2.93% of the aflatoxin is aflatoxin G2 which is only detected in Sango and Agbowo samples (i.e., 1.19 and 2.65 μg/kg, resp.). PMID:27092289

  13. Streptomyces-Aspergillus flavus interactions: impact on aflatoxin B accumulation.

    Science.gov (United States)

    Verheecke, C; Liboz, T; Anson, P; Zhu, Y; Mathieu, F

    2015-01-01

    The aim of this work was to investigate the potential of Streptomyces sp. as biocontrol agents against aflatoxins in maize. As such, we assumed that Streptomyces sp. could provide a complementary approach to current biocontrol systems such as Afla-guard(®) and we focused on biocontrol that was able to have an antagonistic contact with A. flavus. A previous study showed that 27 (out of 38) Streptomyces sp. had mutual antagonism in contact with A. flavus. Among these, 16 Streptomyces sp. were able to reduce aflatoxin content to below 17% of the residual concentration. We selected six strains to understand the mechanisms involved in the prevention of aflatoxin accumulation. Thus, in interaction with A. flavus, we monitored by RT-qPCR the gene expression of aflD, aflM, aflP, aflR and aflS. All the Streptomyces sp. were able to reduce aflatoxin concentration (24.0-0.2% residual aflatoxin B1). They all impacted on gene expression, but only S35 and S38 were able to repress expression significantly. Indeed, S35 significantly repressed aflM expression and S38 significantly repressed aflR, aflM and aflP. S6 reduced aflatoxin concentrations (2.3% residual aflatoxin B1) and repressed aflS, aflM and enhanced aflR expression. In addition, the S6 strain (previously identified as the most reducing pure aflatoxin B1) was further tested to determine a potential adsorption mechanism. We did not observe any adsorption phenomenon. In conclusion, this study showed that Streptomyces sp. prevent the production of (aflatoxin gene expression) and decontamination of (aflatoxin B1 reduction) aflatoxins in vitro. PMID:25632796

  14. miR-20b, miR-98, miR-125b-1*, and let-7e* as new potential diagnostic biomarkers in ulcerative colitis

    DEFF Research Database (Denmark)

    Coskun, Mehmet; Bjerrum, Jacob Tveiten; Seidelin, Jakob Benedict;

    2013-01-01

    and controls were found. The miRNAs with the strongest differential power were identified (miR-20b, miR-99a, miR-203, miR-26b, and miR-98) and found to be up-regulated more than a 10-fold in active UC as compared to quiescent UC, CD, and controls. Two miRNAs, miR-125b-1* and let-7e*, were up-regulated more...... than 5-fold in quiescent UC compared to active UC, CD, and controls. Four of the seven miRNAs (miR-20b, miR-98, miR-125b-1*, and let-7e*) were validated by qPCR and found to be specifically upregulated in patients with UC. Using in silico analysis we found several predicted pro-inflammatory target...... genes involved in various pathways, such as mitogen-activated protein kinase and cytokine signaling, which are both key signaling pathways in UC. CONCLUSION: The present study provides the first evidence that miR-20b, miR-98, miR-125b-1*, and let-7e* are deregulated in patients with UC. The level...

  15. Longitudinal metabolic imaging of hepatocellular carcinoma in transgenic mouse models identifies acylcarnitine as a potential biomarker for early detection

    OpenAIRE

    Jadegoud Yaligar; Wei Wei. Teoh; Rashidah Othman; Sanjay Kumar Verma; Beng Hooi Phang; Swee Shean Lee; Who Whong Wang; Han Chong Toh; Venkatesh Gopalan; Kanaga Sabapathy; S. Sendhil Velan

    2016-01-01

    The cumulative effects of hepatic injury due to hepatitis B virus (HBV) infections and aflatoxin-B1 (AFB1) exposure are the major risk factors of HCC. Understanding early metabolic changes involving these risk factors in an animal model closely resembling human hepatocellular carcinoma (HCC) is critical for biomarker discovery and disease therapeutics. We have used the hepatitis B surface antigen (HBsAg) transgenic mouse model that mimics HBV carriers with and without AFB1 treatment. We inves...

  16. The Molecular Epidemiology of Chronic Aflatoxin Driven Impaired Child Growth

    Science.gov (United States)

    Turner, Paul Craig

    2013-01-01

    Aflatoxins are toxic secondary fungal metabolites that contaminate dietary staples in tropical regions; chronic high levels of exposure are common for many of the poorest populations. Observations in animals indicate that growth and/or food utilization are adversely affected by aflatoxins. This review highlights the development of validated exposure biomarkers and their use here to assess the role of aflatoxins in early life growth retardation. Aflatoxin exposure occurs in utero and continues in early infancy as weaning foods are introduced. Using aflatoxin-albumin exposure biomarkers, five major studies clearly demonstrate strong dose response relationships between exposure in utero and/or early infancy and growth retardation, identified by reduced birth weight and/or low HAZ and WAZ scores. The epidemiological studies include cross-sectional and longitudinal surveys, though aflatoxin reduction intervention studies are now required to further support these data and guide sustainable options to reduce the burden of exposure. The use of aflatoxin exposure biomarkers was essential in understanding the observational data reviewed and will likely be a critical monitor of the effectiveness of interventions to restrict aflatoxin exposure. Given that an estimated 4.5 billion individuals live in regions at risk of dietary contamination the public health concern cannot be over stated. PMID:24455429

  17. Theoretical characterization of aflatoxins and their phototoxic reactions

    Science.gov (United States)

    Guedes, Rita C.; Eriksson, Leif A.

    2006-05-01

    Key molecular properties are calculated for the 8 most common aflatoxins at the B3LYP/6-31 + G(d,p) level. Special attention is given the possibility of aflatoxins to generate reactive oxygen species (ROS). It is concluded that the excited triplet states of the aflatoxins have properties that make them very potent ROS generators, in addition to direct photoinduced addition reactions. The elevated toxicity of aflatoxin B1 is discussed in terms of its lower ionization potential, and the coincidence of higher lying triplet states with dominant low-lying singlet excitations, which enables rapid intersystem crossing and decay along the triplet channel to the T 1 state.

  18. Use of Cold Atmospheric Plasma to Detoxify Hazelnuts from Aflatoxins

    Directory of Open Access Journals (Sweden)

    Ilenia Siciliano

    2016-04-01

    Full Text Available Aflatoxins, produced by Aspergillus flavus and A. parasiticus, can contaminate different foodstuffs, such as nuts. Cold atmospheric pressure plasma has the potential to be used for mycotoxin detoxification. In this study, the operating parameters of cold atmospheric pressure plasma were optimized to reduce the presence of aflatoxins on dehulled hazelnuts. First, the effect of different gases was tested (N2, 0.1% O2 and 1% O2, 21% O2, then power (400, 700, 1000, 1150 W and exposure time (1, 2, 4, and 12 min were optimized. In preliminary tests on aflatoxin standard solutions, this method allowed to obtain a complete detoxification using a high power for a few minutes. On hazelnuts, in similar conditions (1000 W, 12 min, a reduction in the concentration of total aflatoxins and AFB1 of over 70% was obtained. Aflatoxins B1 and G1 were more sensitive to plasma treatments compared to aflatoxins B2 and G2, respectively. Under plasma treatment, aflatoxin B1 was more sensitive compared to aflatoxin G1. At the highest power, and for the longest time, the maximum temperature increment was 28.9 °C. Cold atmospheric plasma has the potential to be a promising method for aflatoxin detoxification on food, because it is effective and it could help to maintain the organoleptic characteristics.