Protein aflatoxin

Aminobiphenol protein adducts (usually serum proteins) Aflatoxin Br/V7-guaninc adducts in urine... 

Pea.nuts, The proteins of peanuts are low in lysine, threonine, cystine plus methionine, and tryptophan when compared to the amino acid requirements for children but meet the requirements for adults (see Table 3). Peanut flour can be used to increase the nutritive value of cereals such as cornmeal but further improvement is noted by the addition of lysine (71). The trypsin inhibitor content of raw peanuts is about one-fifth that of raw soybeans, but this concentration is sufficient to cause hypertrophy (enlargement) of the pancreas in rats. The inhibitors of peanuts are largely inactivated by moist heat treatment (48). As for cottonseed, peanuts are prone to contamination by aflatoxin. FDA regulations limit aflatoxin levels of peanuts and meals to 100 ppb for breeding beef catde, breeding swine, or poultry 200 ppb for finishing swine 300 ppb for finishing beef catde 20 ppb for immature animals and dairy animals and 20 ppb for humans. 

Test preparation for both methods is similar. It comprises an extraction by a water-acetone mixture with its further degreasing protein isolation and redistribution of aflatoxin B into chloroform, concentrating by means of evaporation of the dissolvent on the rotary evaporator (t = 40°C). 

Toxic compounds polychlorinated biphenyls, polycyclic aromatic hydrocarbons, organochlorine pesticides, chlorinated pesticides, dioxins, veterinary drug residues, hormone residues, aflatoxins, toxic compounds in shellfish. Compoimds of nutritional significance in foods vitamins, fat, lipids, carbohydrates, protein, energy-calorific value, proximates, dietary fibre, ash. Other compounds hormones in blood serum... 

Induced mutagenesis in Escherichia coli is an active process involving proteins with DNA replication, repair, and recombination functions. The available evidence suggests that mutations are generated at sites where DNA has been damaged and that they arise via an error-prone repair activity. In an attempt to understand what specific contributions to mutagenesis are made by DNA lesions, we have studied the mutational specificity of some carcinogens, such as benzo[a]pyrene and aflatoxin, whose chemical reactions with DNA are... 

Aflatoxin Bi (AFB) is a mold metabolite which has been observed to be acutely toxic and carcinogenic to a wide variety of animals (5,6) and has been implicated in human primary hepatic carcinoma (7, 8). Diets deficient in protein have been reported to increase the susceptibility of mammals to acute AFB toxicity and the induction of cancer (2, 9, 10, 11, 12, 13). Increased dietary proteins have increased the carcinogenic activity of AFB fed to rats (1 4) and trout (15.). Supportive of this latter finding has been the reported direct relationship between dietary protein content and AFB-DNA adduct formation in vivo in rats (16, 17). 

A further remarkable finding in the hydrolysis of aflatoxin B1 exo-8,9-epoxide is the relative instability of the dihydrodiol, which under basic conditions exists in equilibrium with an aflatoxin dialdehyde, more precisely a furofuran-ring-opened oxy anionic a-hydroxy dialdehyde (10.134, Fig. 10.30). The dihydrodiol is the predominant or exclusive species at pH < 7, whereas this is true for the dialdehyde at pH >9, the pK value of the equilibrium being 8.2 [204], The dialdehyde is known to form Schiff bases with primary amino groups leading to protein adducts. However, the slow rate of dialdehyde formation at physiological pH and its reduction by rat and human aldo-keto reductases cast doubts on the toxicological relevance of this pathway [206]. 

Jayashree, T., Praveen-Rao, J., and Subramanyam, C. (2000). Regulation of aflatoxin production by Ca2- -/calmodulin-dependent protein phosphorylation and dephosphorylation. FEMS Microbiol. Lett. 83, 215-219. 

Immunotechniques have recently been developed to detect food contaminants, e.g., toxins, growth hormone, antibiotics, pesticides, and herbicides. Penicillin (62) in milk, aflatoxins and mycotoxins (63, 64, 65) in milk, cheeses, yogurt, corn have been detected by immunosensors. Characteristics of protein and receptors in or on the cell surface were used in detecting pathogens such as Listeria and Salmonella by immunosensors (11, 66). The principle of immunosensors has also been applied in pesticide determinations (67, 68). 

Aflatoxins are potent carcinogenic, mutagenic and teratogenic metabolites produced by molds. The major food affected with aflatoxins are corn, peanuts, rice, cottonseeds, dried fruit and milk from ingestion (103). The US action standards established by FDA are 20 pg/Kg for foods consumed by humans and 0.5 pg/kg for milk. In the case of animal feed, the levels are from 100 to 300 pg/kg. Therefore, assays capable of detecting at these levels have to be developed, (see Table 1 (104,105)). Detection of aflatoxins entails conjugation of these small molecules with carrier proteins like bovine serum albumin to produce antibodies (20). A number of commercial kits for aflatoxins are available (see sections on kits and immunoaflinity purification). 

Ingredients used to make duck diets are similar to those used in chicken diets, except that groundnut meal should be avoided because it is often contaminated with aflatoxin (a mycotoxin). Ducks are highly sensitive to this aflatoxin, particularly when diets are low in protein. In addition, canola meal may have to be used at low levels only because ducks are more sensitive to erucic acid and goitiogens than are chickens. 

The most common source of aflatoxins is moldy food, particularly nuts, some cereal grains, and oil seeds. The most notorious of the aflatoxins is aflatoxin B1( for which the structural formula is shown in Figure 19.1. Produced by Aspergillus niger, it is a potent liver toxin and liver carcinogen in some species. It is metabolized in the liver to an epoxide (see Section 7.3). The product is electrophilic with a strong tendency to bond covalently to protein, DNA, and RNA. Other common aflatoxins produced by molds are those designated by the letters B2, G1( G2, and M,. 

Hayes (ZZ) observed that in protein-deficient animals, aflatoxin B was more acutely toxic and not as carcinogenic. 

Figure 5 represents the in vitro metabolism of AFBi by rats fed diets containing either 5T or 20% casein. The production of the hydroxylated metabolites AFQi and AFMj is depressed in animals fed the low protein diet, the depression of metabolism is similar to that seen with the N-demethylation of ethylmorphine. An exception to the depressed production of AFMi and AFQi is the enhanced production of an unidentified aflatoxin, tentatively termed AFXi- This metabolite is retained at the origin on TLC... 

Figure 5. The effect of dietary protein levels on the metabolism of aflatoxin to hydroxylated metabolites. Rats were fed semifurified diets consisting of either 5 casein or 20 casein as the protein source.

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