scholarly journals Application of Immunoaffinity Column Cleanup to Aflatoxin M1 Determination and Survey in Cheese

1996 ◽  
Vol 59 (9) ◽  
pp. 1011-1013 ◽  
Author(s):  
S. DRAGACCI ◽  
J. M. FREMY

Milk products such as cheeses may be contaminated by aflatoxin M1 when manufactured with milk from dairy cattle that have consumed aflatoxin B1-contaminated feeds. The usefulness of immunoaffinity columns to determine aflatoxin M1 content in many kinds of cheeses with very good recoveries is demonstrated. The analysis of aflatoxin M1 in a 1990 to 1995 limited survey in France shows that the occurrence of this mycotoxin in cheeses is rather infrequent. With the exception of samples from 1989 to 1990 when aflatoxin B1-contaminated maize meals were incidentally imported to supplement dairy cattle feed, very few samples were found with above 0.200 μg of aflatoxin M1 per kg of cheese, the maximum acceptable level.

Aflatoxin M1 is one of mycotoxin derivatives, which is secreted in milk of dairy cattle fed on feed contaminated with Aflatoxin-B1 (AFB1). The current study was designed to prepare a vaccine against AFB1and to evaluate its efficacy in reducing or preventing secretion of AFM1 in milk. Aflatoxin-B1 was prepared, purified and transformed into oxime, then it was fixed on bovine serum albumins. The AFB1-BSA conjugate was adjuvanted with Gold Nano particles then Montanide ISA 206. The prepared vaccine was used for immunization of rabbits by S/c routes as 100 µg/dose and dairy cattle by I/M routes as 500 µg/dose. The vaccinated animals were boosted at 3 weeks post primary immunization. Serum samples were collected and examined for the anti-AFB1 using AGPT. A mean titer of 15.2 AGPU/ml was detected at 2 weeks post primary vaccination then significantly increased till reached to 76.8 AGPU/ml at 6 weeks post Booster vaccination. All vaccinated rabbits were challenged with dose of 0.3 mg AFB1 toxin/Kg. The vaccinated rabbit showed 100% protection and no AFB1 toxin residue was detected in their livers. Milk samples were collected from non-vaccinated and AFB1-immunized dairy cattle then examined with ELISA for quantitation of AFM1 residues before and after vaccination. The results showed that the prepared AFB1 vaccine was safe, potent and able to reduce AFM1 release in milk of vaccinated heifers by 70%. So the vaccination of lactating animals with the AFB1vaccine might represent a valid tool for the prevention of AFM1 contamination of milk and dairy products.


2021 ◽  
Vol 26 (4) ◽  
pp. 2759-2764
Author(s):  
DRAGAN GLAMOČIĆ ◽  
MIROSLAVA POLOVINSKI HORVATOVIĆ ◽  
IGOR JAJIĆ ◽  
SAŠA KRSTOVIĆ ◽  
MIRKO IVKOVIĆ ◽  
...  

Nutrition of dairy cattle is based on two components, concentrates and forages. The main forages in Vojvodina, north province of Serbia is silage made from the whole plant of corn. After the outbreak of aflatoxin B1 in corn in 2012, the occurrence of aflatoxin B1 in corn as a source of contamination of aflatoxin M1 in milk was very broadly investigated. There is no data regarding the occurrence of aflatoxin B1 in silage and how much silage can contribute to the overall intake of aflatoxin B1 in this region. This work is an attempt to estimate how much silage, in condition and practice used in Vojvodina, contributes to the intake of aflatoxin B1, and consequently aflatoxin M1 in milk. In total, 82 samples of corn grain and 72 samples of corn silage were analyzed on the occurrence of aflatoxin B1 during 2017-2018 period. Aflatoxin B1 was found in 13.41% of corn samples in the range from 6.82 to 187.5 ppb (average 63.5 ppb). All positive samples were from 2017, while no positive samples were found during 2018. Incidence of aflatoxin B1 in silage was 54.17% in the range of 3.5-58.0 ppb (12% moisture content) or 0.95-16.1 ppb in the fresh matter. Results suggest that silage can be a significant factor to overall intake of aflatoxin B1 and that further research is needed.


1985 ◽  
Vol 48 (1) ◽  
pp. 11-15 ◽  
Author(s):  
RALPH L. PRICE ◽  
J. H. PAULSON ◽  
OTIS G. LOUGH ◽  
CONRAD GINGG ◽  
ANDY G. KURTZ

Whole cottonseed determined to have aflatoxin B1 (AFB1) levels of 5, 31, 104, 280, and 560 μg/kg was fed as 15% of the total dairy ration to a commercial herd of 90 grade Holstein dairy cattle for 70 d. Milk from the bulk tank was sampled either daily or after each milking and analyzed for aflatoxin M1 (AFM1). The ratio for AFB1 in the dairy ration to AFM1 in the milk averaged 75 to 1 under conditions and at levels tested with no consistent relation to the level of AFB1 in the feed. Approximately 1.6% conversion occurred during the steady state of consumption and secretion. The federal action level of 0.5 μg AFM1/L of milk would be produced by cows consuming a ration containing 15% whole cottonseed contaminated with approximately 250 μg AFB1/kg.


2019 ◽  
Vol 13 (2) ◽  
pp. 19-23
Author(s):  
Masoumeh Mahmoodi Maymand ◽  
◽  
Mansooreh Mazaheri ◽  
Mahboobeh Talebi Mehrdar ◽  
◽  
...  

Background: Mycotoxins are the secondary metabolites of molds and have adverse effects on humans, animals, and crops, resulting in illnesses and economic losses. Aflatoxin M1 (AFM1) is a hepatocarcinogen found in the milk from animals that have consumed feeds contaminated with aflatoxin B1 (AFB1). Milk is a highly nutritious food and is a source of necessary macro- and micro-nutrients for the growth, development and maintenance of human health. Methods: The presence of AFM1 was investigated in 70 samples of imported pasteurized and powdered milk products available to the Iranian consumers. The level of AFM1 was determined by HPLC method equipped with immunoaffinity cleanup. Results: The results showed that 32% of the analyzed samples were positive for AFM1 at 0.05-3.31 μg/kg. Also, 16% of analyzed samples were positive for AFM1at concentrations higher than the limit permitted by the Iranian standards. Conclusion: The detection of AFM1 contamination in the analyzed samples indicates the importance of the health of animal feeds. Thus, monitoring the imported feed materials, especially those arriving at Iranian borders is crucial in the prevention of AFM1 and AFB1 contaminations spreading across the domestic market.


2019 ◽  
Vol 10 (1) ◽  
pp. 3-49
Author(s):  
Sandra Jakšić ◽  
Milica Živkov Baloš ◽  
Jasna Prodanov Radulović ◽  
Igor Jajić ◽  
Saša Krstović ◽  
...  

Aflatoxin M1 (AFM1) is a hepatocarcinogenic derivative of aflatoxin B1 excreted into the milk after ingestion of contaminated feed. The presenceof AFM1 in milk and milk products is of huge concern for human health. In this paper, the results on long term assessment of AFM1 in milk produced in Serbia are presented. In the period 2013 to 2016, 427 milk samples were examined for AFM1. In 34.4 % of samples, the content of AFM1was higher than 0.05 μg/kg. The article also offers a review of the fate of aflatoxin in milk products during the different operations in milk processing. The evaluation of the influence of processing on AFM1 stability can propose economic strategy for resolving cases of accidents due to AFM-1contamination of milk.


1982 ◽  
Vol 45 (8) ◽  
pp. 752-777 ◽  
Author(s):  
RHONA S. APPLEBAUM ◽  
ROBERT E. BRACKETT ◽  
DANA W. WISEMAN ◽  
ELMER H. MARTH

Aflatoxins are toxic and carcinogenic secondary metabolites produced by some common aspergilli during growth on feeds, foods or laboratory media. Aflatoxin B1 (AFB1) is a decaketide (C20-polyketide) which is synthesized by the mold from acetate units via the polyketide pathway. Methionine contributes the methoxy-methyl group. Six known intermediate compounds in the biosynthesis of AFB1 include norsolorinic acid, averantin, averufin, versiconal hemiacetal acetate, versicolorin A and sterigmatocystin. Other aflatoxins (B2, B2a, G1, G2 and G2a) appear to be conversion products of AFB1. When aflatoxins, and in particular AFB1, occur in feed and are consumed by dairy cattle, a variety of symptoms can occur, which includes unthriftiness, anorexia and decreased milk production. Changes in amounts of enzymes and other blood constituents also result from ingestion of AFB1. The hepatic microsomal mixed-function oxidase system of the cow converts some of the ingested AFB1 into aflatoxin M1 (AFM1), which is excreted in milk. AFM1 retains the toxicity of, but is less carcinogenic than AFB1. Certain heat treatments associated with milk processing appear to inactivate a portion of the AFM1 in milk. If raw milk contains AFM1, products (fluid products, nonfat dried milk, cultured milks, natural cheese, process cheese, butter) made from such milk also will contain AFM1. AFM1 appears to be associated with the casein fraction of milk, hence concentrating the casein in the manufacture of products (e.g. cheese, nonfat dry milk) is accompanied by concentrating of the AFM1. Methods involving thin-layer or high-performance liquid chromatography are commonly used to detect and quantify AFM1 in milk and milk products.


2003 ◽  
Vol 86 (6) ◽  
pp. 1179-1186 ◽  
Author(s):  
Joerg Stroka ◽  
Christoph von Holst ◽  
Elke Anklam ◽  
Matthias Reutter ◽  
A Barmark ◽  
...  

Abstract A collaborative study was conducted to evaluate the effectiveness of an immunoaffinity column cleanup liquid chromatography (LC) method for determination of aflatoxin B1 in cattle feed at a possible future European regulatory limit (1 ng/g). The test portion was extracted with acetone–water (85 + 15), filtered, diluted with water, and applied to an immunoaffinity column. The column was washed with water to remove interfering compounds, and the purified aflatoxin B1 was eluted with methanol. Aflatoxin B1 was separated and determined by reversed-phase liquid chromatography (RP–LC) and detected by fluorescence after post column derivatization (PCD) involving bromination. PCD was achieved with either pyridinium hydrobromide perbromide (PBPB), used by 14 laboratories, or an electrochemical cell and addition of bromide to the mobile phase, used by 7 laboratories. Both derivatization techniques were not significantly different when compared by the t-test; the method was statistically evaluated for all laboratories together (bromination and PBPB). The cattle feed samples, both spiked and naturally contaminatedwithaflatoxinB1, were sent to 21 laboratories in 14 different countries (United States, Japan, and Europe). Test portions were spiked at levels of 1.2 and 3.6 ng/g for aflatoxin B1. Recoveries ranged from 74 to 157%. Based on results for spiked samples (blind pairs at 2 levels) as well as naturally con-taminated samples (blind pairs at 3 levels), the relative standard deviation for repeatability (RSDr) ranged from 5.9 to 8.7%. The relative standard deviation for reproducibility (RSDR) ranged from 17.5 to 19.6%. The method showed acceptable within-and between-laboratory precision for this matrix, as evidenced by HORRAT values, at the target levels of determination for aflatoxin B1. No major differences in RSD were observed, showing that the composition of the feeds was not a factor for the samples tested and that the method was applicable for all materials used.


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