BACKGROUND: Aflatoxin M1 (AFM1) is the main monohydroxylated derivative of aflatoxin B1 (AFB1) formed in liver and excreted into milk. AFM1  creates  certain hygienic risks for human health. OBJECTIVES: The aim of this study was to determine AFM1 level in raw milk samples in Yazd province. METHODS: This investigation was a descriptive-cross sectional study. Eighty raw milk samples were collected from four cities (Yazd, Taft, Mehriz and Sadogh) in Yazd province in winter and spring seasons. The concentration of AFM1 was determined by ELISA method. The analysis of the results was performed using ANOVA and Chi-square tests. RESULTS: All samples (100%) were contaminated with AFM1, with the concentrations ranging from 3.18 to 92.24 ng/l with a mean concentration of 22.07 ng/l. AFM1 level in 13.7% of raw milk samples was higher than the maximum tolerance limit of 50 ng/l accepted by the European Union (EU). The contamination level of AFM1 in winter samples (28.21 ng/l) was higher than spring samples (15.92 ng/l). Also, the highest and lowest contamination levels were observed in milk samples collected from Sadogh (mean 42.21 ng/l) and Yazd (12.79 ng/l) cities, respectively. CONCLUSIONS: Results demonstrated AFM1 was detected with a mean concentration of 22.07 ng/l in milk samples of Yazd province. Moreover, 13.7% of samples contained AFM1 at hazardous levels for human health.

Hydrated sodium calcium aluminosilicate (HSCAS), an anticaking agent for agricultural feeds, was added to aflatoxin (AF)-contaminated diets of 3 lactating dairy cows and evaluated for its potential to reduce aflatoxin M1 (AFM1) residues in milk. During phase I, cows were fed alternating diets that consisted of 200 microgram of AF/kg of feed for 7 days, 0.5% HSCAS plus 200 microgram of AF/kg of feed for 7 days, and feed with the HSCAS removed for a final 7 days. The AFM1 milk concentrations from the intervals with HSCAS added to diets were compared with those times when HSCAS was absent. The presence of 0.5% HSCAS in feed containing 200 microgram of AF/kg reduced AFM1 secretion into the milk by an average of 0.44 microgram/L (from pretreatment of 1.85 microgram/L to 1.41 microgram/L with HSCAS, a 24% reduction). Following a 10-day period of noncontaminated feed consumption and no AFM1 residues in the milk, phase II of the study was begun. The same experimental design as phase I was used, but the dosages of HSCAS and AF were changed to 1.0% and 100 microgram/kg of feed, respectively. The addition of 1.0% HSCAS in feed containing 100 microgram of AF/kg decreased AFM1 content in the milk by an average of 0.40 microgram/L (from a pretreatment of 0.91 microgram/L to 0.51 microgram/L when HSCAS was present, a 44% reduction). These findings suggest that HSCAS, a high-affinity sorbent compound for AF in vitro, is capable of reducing the secretion of AFM1 into milk.

A study was conducted to determine aflatoxins in tissues and non-tissues of 2 Holstein cows given oral doses of 0.35 mg of purified aflatoxin B1/kg of body weight/day for 3 consecutive days. Cow 1 was slaughtered 24 hours after the 3rd dose, and cow 2, after day 3, was fed aflatoxin-free rations for 7 additional days before slaughter. Tissue samples of brain, gallbladder and bile, heart, intestine, kidney, liver, lung, mammary gland, skeletal muscle, spleen, supramammary lymph nodes, thymus, and tongue, and nontissue samples of blood, feces, milk, rumen content, and urine were examined. Aflatoxins B1 and M1 were found in all samples of cow 1, except the thymus. Kidney, liver, and mammary gland had the highest concentrations of total aflatoxins (57.9, 13.2, and 25.1 ng/g, respectively), with the aflatoxin M1 concentration 40 times more than the aflatoxin B1 level in kidney. Aflatoxin residues were present (0.02 to 0.11 ng/g) only in kidney, liver, and intestine of the tissues from cow 2 (fed aflatoxin-free feed for 7 additional days). Aflatoxin B1 was not present in nontissue samples, but aflatoxin M1 (0.10 and 1.5 ng/ml) was found in the last milk and urine samples from the same cow. Urine assays are a possible way to monitor the presence of aflatoxin residues in meat tissues.

Two of 3 groups of Holstein-Friesian steers (groups II and III; n = 5 each) were fed a ration containing corn naturally contaminated with 800 ng of aflatoxin/g. The other group of steers (group I; n = 5) was fed a ration containing noncontaminated corn. The respective rations were fed for 17.5 weeks, except the ration given to group III; the latter's first diet (contaminated with aflatoxin) was changed to a noncontaminated diet after 15 weeks, continuing for the remaining 2.5 weeks. All steers were killed and tissues and fluids were obtained for aflatoxin analysis. Although aflatoxin B1 and M1 could be detected in blood and urine at several sampling times during the experimental period in groups II and III steers (given the diets containing aflatoxin), there appeared to be no effects on body weight gains and immune phenomena, such as lymphoblastogenesis and antibody production, but there was a waning of the delayed cutaneous hypersensitivity in steers given aflatoxin-contaminated diets. In group III animals (diet was changed to noncontaminated ration at 15 weeks), aflatoxin B1 and M1 disappeared from urine before they were slaughtered. All tissues and fluids, except the rumen contents from these group III steers, were void of detectable aflatoxins B1 and M1 at necropsy. The concentrations of aflatoxin B1 in the rumen content of the latter steers were low. All tissues collected at necropsy from the group II steers fed the aflatoxin diet throughout the 17.5 weeks had detectable aflatoxins B1 or M1 present.

This work was planned to qualitatively assess AFM1 residues in milk as well as milk product samples. Cow’s raw milk, Aged Roomy cheese, Domiati and Kariesh cheese (50 of each type) were collected from various supermarkets in Mansoura governorate in Egypt during 2020-2022. Samples were tested using the TLC method, positive samples from TLC method were exposed to the HPLC method for detection of their levels and their concentrations were compared with the permissible limits. The obtained results showed high incidence in raw cow milk (36%) followed by fresh Kariesh cheese (28%), then aged roomy cheese (24%), and then Damietta cheese  (12%) collected from the local markets in Mansoura by TLC. The obtained concentrations in analyzed dairy samples were compared with the Permissible Limits (PL) set by the concerning agents [US, EU] by quantitative detection using HPLC. The detectable concentration of AFM1 in raw milk and cheese by HPLC showed high concentration of aflatoxin M1 in aged roomy cheese  (121.08) followed by raw cow milk  (80.83), followed by fresh Kariesh cheese  (50.14), and then Damietta cheese  (35.33) collected from the local markets in in Dakahlia governorate. The current study's findings showed that raw milk and cheese sold in Mansoura city are occasionally contaminated with AFM1. As a result, it is strongly advised to effectively heat treat raw milk and give animals nutrition free of aflatoxins.

Enzyme-linked immunosorbent assay screening of antibody produced against aflatoxin was accomplished by a new and simple procedure. To demonstrate the new indirect ELISA technique used, antibody against aflatoxin M1 was produced in female BALB/CJ mice by immunization with an aflatoxin M1-bovine serum albumin conjugate. Instead of coating test-plate wells with purified antibody (direct ELISA) or synthesizing a second protein-aflatoxin conjugate (aflatoxin M1-poly-L-lysine) to coat test-plate wells, wells were coated with the readily available aflatoxin M1-bovine serum albumin and aflatoxin B1-bovine vine serum albumin. This method, applicable for any aflatoxin conjugated by the common cyclopentano-carboxymethoxyl-oxime technique, eliminates the more time-consuming and technically difficult portions of earlier direct and indirect ELISA. The new technique can be valuable in continued efforts toward development of new and improved immunoassays against aflatoxin metabolites.