Survey of total aflatoxins in camels sera by enzyme linked immunosorbent assay (ELISA)

A survey of total aflatoxins was carried out on samples of camels blood sera in order to assess aflatoxin hazards in camels following some deaths among animals during July through September, 1990 in Al Ain area, about 16 km from Abu Dhabi City. Epidemiological and laboratory based studies indicated that the outbreak was associated with the consumption of mould damaged feed.A flavus andA parasiticus and varying amount of aflatoxin were detected in samples tested. 53 samples of camel sera were collected for the analysis: 16 out of 53 samples were collected from apparently clinically healthy racing camels. All these samples were examined for total aflatoxins. All of the 16 samples were found to contain aflatoxins ranging from 5 to 50ng/mL total aflatoxin and 28 out of the 37 samples were found to contain aflatoxin at the range of 2 to 12 pg/mL total aflatoxins.     

Aflatoxin contamination of maize in flooded areas of Bhagalpur, India.

Maize in flood-affected areas in the Bhagalpur district of India in 1985 demonstrated heavy infestations of Aspergillus flavus and aflatoxins. Sixty samples collected from various field lots were positive in the bright greenish yellow fluorescence test. However, only 42 of those samples were found to be contaminated with deleterious levels of aflatoxins. Factors affecting aflatoxin contamination are discussed.     

Reduction of mutagenic activity of aflatoxins after UV-irradiation.

Solutions of aflatoxins B1, B2, G1 and G2 and solid films of aflatoxin B1 were irradiated with UV light of wavelength corresponding to the absorption band of aflatoxins at approximately 365 nm (approximately 27 500 cm-1). The reaction was followed spectrophotometrically. Simultaneously the mutagenicity of samples of aflatoxins was determined using Salmonella typhimurium/mammalian microsome test. UV-irradiation caused a modification in all aflatoxins studied (in aqueous and nonaqueous solutions as well as in a solid film); the final photoproducts lost completely the mutagenic activity.     

Occurrence of Aflatoxins and Aflatoxin-Producing Strains of Aspergillus spp. in Soybeans

Above average rainfall in Maryland during August, September, and October 1971 resulted in heavy mold growth in soybeans while still in the field. Of 28 samples of soybean seed, aflatoxins were found in 14, 2 of which had been used in poultry feed. Aflatoxins were identified by thin-layer chromatography, spectrophotometry, and chicken embryo bioassay. Aspergillus spp. were isolated from 11 samples, and 5 of these isolates produced aflatoxins when grown in liquid culture.     

Detection and estimation of aflatoxins using both chemical and biological techniques

Production of aflatoxins on both natural (rice and corn) and semisynthetic (YES) media was conducted using an identified toxin-producing strain ofAspergillus flavus. TheA flavus strain was able to produce 4 types of aflatoxins, namely B₁, B₂, G₁, and G₂ on rice, corn, and YES media. Quantitative data showed that the concentrations of aflatoxins B₁ and G₁ produced were 52, 40.3, and 39.6; and 64.7, 45.0, and 58.0jug for 50g of rice, corn, and YES media, respectively. In comparison, the yielded amounts of aflatoxins B₂ and G₂ were much lower: 11.5, 17.9, and 17.5; and 28.S, 40.3, and 39.5 μg for 50 g of rice, corn, and YES media, respectively. A bioassay was conducted using the following 5 standard bacterial strains:Bacillus megaterium. Bacillus subtilis, Streptococcus faecal is, Staphylococcus epidermidis, andParacoccus denitrificans as well as a field strain of Candida albicans. All strains exceptP denitrificans showed varied degrees of inhibition when applied with crude aflatoxins at 5 to 40μg/mL. The minimum concentration of crude aflatoxins needed to inhibitP denitrificans was 10μg/mL. Moreover,Candida albicans was not inhibited at any concentration of aflatoxins applied in this work. Both undiluted and diluted (1/10, 1/100, and 1/1000) bacterial broth cultures showed a direct relationship between the diameter of inhibition zones and the concentrations of crude aflatoxins. Mean diameters of (7.0–20.5), (5–14), (4.5–13.0), (3.0–12.0), and (1.5–11.0) mm were observed when various concentrations of aflatoxins were applied usingB megaterium, S epidermidis, S faecal is, B subtilis, andP denitrificans, respectively. Field trials were applied to testify the validity of our data. A 1/100 dilution was prepared from each strain of 4 different species to estimate aflatoxins in samples of contaminated corn. Both chemical and biological assays were carried out at the same time. Data revealed that the most sensitive organism inhibited by as low as 7.5μg aflatoxins/mL wasB megaterium giving an inhibition zone of 10.5 mm, followed byS epidermidis with an inhibition zone of 7.5mm. In relation, the other 2 organisms were less sensitive to crude aflatoxins. Similarly, the biological assay was applied to detect aflatoxins in some samples of wheat, corn, peanut, rice, and poultry rations. Of the 14 wheat and 10 corn samples, only 4 wheat and 2 corn samples were found to be positive. The same results were obtained using TLC analysis.     

Aflatoxin contamination in different fractions of rice from Pakistan and estimation of dietary intakes

The purpose of this study was to determine the distribution of aflatoxins in rice milling fractions and to estimate dietary intakes. A total of 413 rice samples (paddy 58, parboiled 69, brown 84, white 93, and broken 109) were analyzed by HPLC with fluorescence detector. The results showed that 64?% paddy (16.35?±?1.67???g/kg), 38?% parboiled (14.20?±?2.04???g/kg), 33?% brown (9.85?±?1.25???g/kg), 42?% white (7.10?±?1.39???g/kg), and 50?% broken (8.5?±?1.71???g/kg) rice samples were contaminated with aflatoxins. It was found that paddy rice was most contaminated with aflatoxins while white rice was least contaminated. The percentage of samples exceeding EU maximum contents for total aflatoxins in rice (4???g/kg) varied from 14 to 36?% when compared to the analyzed rice fractions. The total estimated amount of aflatoxin intake for average rice consumers ranged from 19.1 to 26.6?ng/kg body weight/day, much higher than the reference value of 1?ng/kg body weight/day. This is the first report discovering that rice is a major contributor to the dietary intake of aflatoxins in Pakistan.     

Aflatoxins isolated by immunoaffinity chromatography from foods consumed in The Gambia, West Africa.

An aflatoxin-specific, monoclonal antibody-based immunoaffinity chromatography method has been developed for the rapid isolation of aflatoxins from human foods. Aflatoxins were isolated by immunoaffinity chromatography from a variety of cooked foods, including maize, rice, millets, groundnut sauces, and leaf sauces, collected in The Gambia, West Africa. The aflatoxins were measured by direct fluorescence or high-pressure liquid chromatography. The highest levels were found for groundnut sauces, mean 162 ppb (range 18 to 943 ppb) for 18 positive samples, but aflatoxins were found in other foods; e.g., maize, mean 9.7 ppb (range 2 to 35 ppb) for nine positive samples. The food analysis results were used with records of the amounts of cooked food to estimate a mean daily intake for an individual of the order of 3.5 micrograms of aflatoxins per day. This approach for exposure assessment is considered in relation to other biomarkers of aflatoxin exposure using biological fluids.     

Fungal flora and aflatoxin contamination of feedstuff samples in Beida Governorate,Libya

Fungi of 19 genera, 30 species, and one variety were isolated from 25 samples of sheep-, cattle- and camel feedstuffs collected from different farms in the Beida Governorate, Libya.Aspergillus, Penicillium andFusarium were the most common genera in the three substrates tested. TLC was used to establish the identity of aflatoxins in the chloroform extract of all samples and the ability to produce aflatoxins byAspergillus flavus in a synthetic liquid medium. Twenty samples out of 25 tested were naturally contaminated and 21 isolates ofA. flavus out of 30 produced at least one of the following aflatoxins: B₁; B₁, G₁; and B₁, B₂, G₁, G₂. This is the first report about the natural occurrence of aflatoxins and aflatoxin-producers of the genusAspergillus in Libya.     

A limited survey of aflatoxins and fumonisins in retail maizebased products in the UK using immunoassay detection

A total of 27 maize-based products destined for human consumption were collected from retail outlets within the city of Glasgow in the UK and were analysed for the presence of aflatoxins using immunoaftinity column chromatography with fluorescence detection and for fumonisins by competitive ELISA. Aflatoxins were detected at a trace level below 4 in eight (30%) of the 27 samples tested, no sample contained aflatoxins at a high level although one sample of sweetcorn did contain aflatoxins at a level of 5-10 Fumonisins were detected in eight (30%) of the samples at levels from 1 to 8mgkg^-1 and a further eight samples contained fumonisin at a level below 1 mgkg^-1 but above the detectable level. The highest concentration of fumonisins was found in a sample of fine corn meal at 8-12mgkg^-1.     

Aflatoxin contamination of groundnuts in Sudan

Groundnut samples, collected soon after harvest, from different districts in the irrigated region (Central Sudan) were free from aflatoxins with the method used. Samples collected from the rainfed region (Western Sudan) showed variable levels of aflatoxin ranging from 100% sample contamination in El Hamdi to only 10% in Casgeal.Damaged pods were highly contaminated with A. flavus and accumulated large amounts of aflatoxins. However, sound intact pods, recorded lower fungal contamination and were almost free of aflatoxins. Groundnut products collected from Khartoum North (Bahri) have higher levels of aflatoxins than those collected from Khartoum and Umdorman. Gray and red roasted pods showed higher amounts of aflatoxins, while the groundnut paste was the least contaminated.None of the three varieties of groundnuts tested in this work was completely resistant to aflatoxin production. A temperature of 30°C and 86.3% relative humidy (RH) are the optimum conditions for both A. flavus growth and aflatoxin production in groundnuts.     

Development and validation of a method for the quantitative determination of aflatoxin contaminants in Maytenus ilicifolia by HPLC with fluorescence detection

A method for the quantification of aflatoxins B1, G1, B2 and G2 in the medicinal herb Maytenus ilicifolia was developed and validated. The method used immunoaffinity columns for sample clean-up and HPLC with fluorescence detection without any derivatisation step. The method showed good inter-day accuracy (bias values in the range 4.5-10.7%) and precision (5-16% RSD) when applied to the determination of levels of aflatoxins ranging from 7 to 20 ppb in the plant material. The detection limits for samples of the plant material spiked with aflatoxins were 3.5 ng/g for B1 and G1 and 0.1 ng/g for B2 and G2. The method was successfully applied to commercial samples of Maytenus ilicifolia for the screening of aflatoxin contaminants.     

A note on the screening of dried shrimps, shrimp paste and raw groundnut kernels for aflatoxin-producing Aspergillus flavus

All of 33 samples of dried shrimps, shrimp paste, peanut butter and raw groundnut kernels were contaminated with fungi. Aspergillus and Penicillium spp. were the predominant types in dried shrimps and raw groundnut kernels but no Aspergillus spp. were present in peanut butter or shrimp paste samples. Among 81 Aspergillus isolates obtained from dried shrimps and raw groundnut kernels, 10 were A. flavus/A. parasiticus, of which five were potential aflatoxin-producing A. flavus strains. No aflatoxins were detected in the food samples although some were visibly mouldy and some had high mould counts. The occurrence of aflatoxin-producing strains of A. flavus in dried shrimps and raw groundnut kernels warrants further investigation of these foods and their products as potentially significant sources of aflatoxins.     

Determination of aflatoxin levels in pistachio and cashew nuts using immunoaffinity column clean-up with HPLC and fluorescence detection

A technique for the detection of aflatoxins in pistachio and cashew nuts using immunoaffinity column clean-up with HPLC and fluorescent detection is presented. Recoveries were in the range of 79–99% for pistachio samples artificially contaminated with 10 g total aflatoxins kg–1 of food sample. For cashew samples recoveries ranged from 80–106%. This method is proposed as an accurate technique for aflatoxin detection in the range of g aflatoxins kg–1 nuts.     

Incidence of aflatoxin producing strains and aflatoxin contamination in dry fruit slices of quinces (Cydonia oblonga Mill.) from the Indian state of Jammu and Kashmir

An investigation was undertaken to obtain data on the occurrence of aflatoxins and the aflatoxin producing potential of Aspergillus flavus strains isolated from dry fruit slices of quinces produced in jammu and Kashmir, India. A total of 147 A. flavus isolates recovered from dr fruit slices were grown in liquid rice flour medium and screened for the production of various aflatoxins by thin layer chromatography. The results showed that 23.14% of the tested isolates were aflatoxigenic, producing aflatoxins B1 and B2 in varying amounts. Aflatoxins G1 and G2 were not detected. All 25 of the investigated market samples were also found to be aflatoxin B1 positive and the level of contamination ranged from 96 to 8164 micrograms/kg of the dry fruit which is quite high in comparison to the permissible level of 30 ppb. As per these results biochemical composition of dry fruit slices of quinces, along with climatic conditions seem to be very favourable for aflatoxin production by the toxigenic A. flavus strains. Therefore, monitoring of aflatoxins in dry fruit slices of quinces is recommended for this region.     

Aflatoxin in corn: ammonia inactivation and bioassay with rainbow trout.

Four samples of corn were compared with respect to their hepatocarcinogenicity in rainbow trout. One corn sample was found by chemical analysis to contain no detectable aflatoxin. A second sample was contaminated with aflatoxins at a level of 180 microgram/kg. Each of the above-mentioned samples was divided, and one-half of each was ammoniated. These four samples were added to a semipurified basal diet and fed to a sensitive strain of rainbow trout. It was found that ammoniation inactivated the aflatoxins and reduced the carcinogenicity of the contaminated corn to a level that was not significantly different from that with the basal control diet. It was also found that the ammoniation process did not reduce the nutritive value of the corn.     

Aflatoxin in corn: ammonia inactivation and bioassay with rainbow trout.

Four samples of corn were compared with respect to their hepatocarcinogenicity in rainbow trout. One corn sample was found by chemical analysis to contain no detectable aflatoxin. A second sample was contaminated with aflatoxins at a level of 180 microgram/kg. Each of the above-mentioned samples was divided, and one-half of each was ammoniated. These four samples were added to a semipurified basal diet and fed to a sensitive strain of rainbow trout. It was found that ammoniation inactivated the aflatoxins and reduced the carcinogenicity of the contaminated corn to a level that was not significantly different from that with the basal control diet. It was also found that the ammoniation process did not reduce the nutritive value of the corn.     

Ultrasensitive nanogold probe-based immunochromatographic assay for simultaneous detection of total aflatoxins in peanuts

An ultrasensitive immunochromatographic (IC) assay for simultaneous detection of total aflatoxins (AFB1, AFB2, AFG1, and AFG2) was developed to meet the requirement for rapidly monitoring aflatoxins in agro-products. The assay was based on a competitive format and its sensitivity was improved by using a novel criterion to screen the optimal amount of monoclonal antibody (MAb) labeled to nanogold particles. The visual detection limits (VDLs) for aflatoxins B1, B2, G1, and G2 in peanut matrix were 0.03, 0.06, 0.12, and 0.25 ng mL(-1), respectively, which were lower than those of published literatures. The results of IC assay were in good agreement with those of high performance liquid chromatography (HPLC) in the analysis of aflatoxins in peanuts, demonstrating the practical applicability of the developed assay in real samples. This qualitative test based on the visual evaluation of results did not require any equipment. Overall, to our knowledge, this is the first report of qualitative detection for total aflatoxins by immunochromatographic assay.     

Contamination of Red Chilli with Aflatoxin B1 in Pakistan

A survey of red chilli (Capsicum indicum) for contamination with aflatoxins was performed on different samples comprising whole, crushed and powdered red chilli collected from various stores located in the city of Karachi, Pakistan. Red chilli required rather rigorous clean-up procedure for removal of adulterants and interference resulting from various types of compounds. A modified Romer method followed by bi-directional thin layer chromatography (TLC) was used for the detection of aflatoxins and confirmatory tests were performed by spraying the TLC plates with 50% sulphuric acid and making the derivative with trifluoroacetic acid. Of all the 176 samples of red chilli examined, 66% were found to be contaminated with aflatoxin B₁. Generally, samples of red chilli exammined were found to be fairly low in aflatoxin B₁ content, whereas only seven samples were found to contain concentrations greater than 25 μg/kg of aflatoxin B₁.     

Ochratoxin A and aflatoxins in breast milk samples from Sierra Leone

Breast milks from 113 mothers in two Under-Five Clinics in the Southern Province of Sierra Leone, namely, Njala and Bo, were examined for their mycotoxin content. Only 10 were mycotoxin-free. Eighty-eight per cent of samples contained various aflatoxins and 35% contained ochratoxin A (OTA). Few samples (15%) had a single mycotoxin. Thirty-six (32%) had two mycotoxins and 50 (40%) had three or more. The occurrence of OTA in combination with various aflatoxins was recorded. It is concluded that infants in Sierra Leone are exposed to OTA and aflatoxins at levels which in some cases far exceed those permissible in animal feed in developed countries.     

Occurrence of aflatoxin in some liver curative herbal medicines

Fifty herbal medicine samples of seven different taxa known to cure liver disorders were analysed for aflatoxin contamination. Twenty-three samples, out of 50, were contaminated with various levels of aflatoxins. Amongst the 23 contaminated samples the maximum level of aflatoxin B¹ recorded was 2.23 μg g^-1 in Asparagus racemosus and the minimum 0.28 μg g^-1 in Emblica officinalis . Aflatoxin G¹ was only found in one species, Terminalia belarica . Aflatoxin production of the isolates of Aspergillus flavus was also examined and the highest levels were produced by isolates from A. racemosus (1.07-2.47 μg ml^-1). Aflatoxin contamination of herbal drugs may be a risk for patients because the level of aflatoxins is much higher than the tolerance level fixed by the WHO for foods.