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Single and combination effects of administering salinomycin and aflatoxin to broiler chicks.

A factorial design (2 by 3) was used to evaluate the interaction between aflatoxin (0, 2.5, and 5 mg per kg) and salinomycin [0, 60 g per ton (909 kg)]. There were four replicates of 10 chicks per treatment. The chicks were maintained in batteries from 0 to 3 wk of age, with feed and water available for ad libitum intake. Aflatoxin at both levels (2.5 and 5 mg per kg) with and without salinomycin decreased body weight. The efficiency of feed utilization was affected only at the 5 mg per kg level of aflatoxin. Feeding 5 mg per kg of aflatoxin alone decreased the hemoglobin level. The inclusion of salinomycin (60 g per ton) in the diet with 2.5 or 5 mg per kg of aflatoxin initiated no significant change in body weight or feed efficiency. No significant interaction was observed between aflatoxin and salinomycin on any of the parameters measured.

Aflatoxins↗

Effect of cold acclimation on the broiler chicks' resistance to dietary aflatoxin.

The effect of acclimation to environmental temperatures of 10 to 12 or 28 to 30 C on the resistance of broiler chicks to dietary aflatoxin was examined. Broiler chicks were acclimated from day-of-age for 2 wk to environmental temperatures of 10 to 12 or 28 to 30 C. On Day 14, a single oral dose of aflatoxin (8 mg per kg of body weight) was administered to 50 chicks in each environment. An increase in aflatoxin resistance, as assessed by survival rate, was conveyed by acclimation to cold temperatures. In each environmental chamber, a separate group of chicks was maintained for 2 additional wk, but those groups received 5 mg of aflatoxin per kg feed. By the end of the study, aflatoxicosis was characterized by: 1) a significant (P less than or equal to .05) decrease in body weight; 2) increases in spleen weight, liver weight, liver lipid, and liver dry-matter content; 3) changes in the serum levels of total protein, albumin, glucose, cholesterol, uric acid, potassium, phosphorus, iron and calcium; and 4) increased hepatic hyperplasia. Acclimation to 10 to 12 C was characterized by: 1) an increase in body weight, liver weight, spleen weight and bursa weight; 2) changes in the serum glucose and potassium levels; and 3) a decrease in glutamic-oxaloacetic transaminase activity. Significant aflatoxin by temperature interactions were evident only in serum levels of glucose and phosphorus, and in the serum activity of glutamic-oxaloacetic transaminase. These data suggest that acclimation to cool temperatures does not play a significant role in the resistance by broiler chickens to chronic aflatoxin exposure.

Adaptation, Physiological↗

Interaction of naturally occurring aflatoxins in poultry feed and immunization against fowl cholera.

A total of 1,175 poultry feed samples originating from different farms were analyzed for aflatoxin. Poor growth rate and reduced egg production were the main complaints. The rate of contamination with aflatoxin ranged from 10 to 54% of all samples. Of samples examined 30.7% proved positive for aflatoxin with a concentration ranging from 1 to 2,000 ppb. Outbreaks of fowl cholera were diagnosed on two farms where aflatoxin was detected in the rations used. The impact of aflatoxin in the feed on the efficacy of immunization against fowl cholera was monitored by a hemagglutination test and the titers of the involved farms were compared with experimental groups fed on aflatoxin-free rations and vaccinated with the same polyvalent fowl cholera bacterin. The antibody titers of the experimental groups were 4 to 15 times higher than those of the involved farms.

Aflatoxin B1↗

Efficacy of hydrated sodium calcium aluminosilicate to reduce the individual and combined toxicity of aflatoxin and ochratoxin A.

A 2 x 2 x 2 factorial arrangement of treatments consisting of dietary aflatoxin (3.5 micrograms/g), ochratoxin A (2.0 micrograms/g), and hydrated sodium calcium aluminosilicate (HSCAS, .5%) was used to evaluate the individual and combined effects of these treatments. There were six replicate pens of 10 broilers per pen for each of the eight treatments. The broilers were maintained on these treatments from 1 day to 3 wk of age with feed and water available for ad libitum intake. Aflatoxin and ochratoxin A each significantly decreased body weight, serum protein, albumin, and cholesterol and increased the relative weight of the liver, kidney, and proventriculus. Aflatoxin increased the relative weight of the heart and decreased serum aspartate aminotransferase activity and ochratoxin A increased serum uric acid. The toxicity resulting from the combination of aflatoxin and ochratoxin A was more severe than when either of these mycotoxins were present alone. Addition of HSCAS alone did not alter any of the parameters evaluated. The HSCAS reduced the toxicity of aflatoxin, but had little effect on either the toxicity of ochratoxin A alone or the toxicity resulting from the combination of aflatoxin and ochratoxin A.

Aluminum Silicates↗

Occurrence of aflatoxins, fumonisin B1, and zearalenone in foods and feeds in Botswana.

Sorghum and maize form the main dietary staple foods in Botswana. Other products such as peanuts, peanut butter, phane (an edible larval stage of an emperor moth Imbrasia belina Westwood), and pulses (cowpeas and beans) are also widely used as food and for the manufacture of feeds. These important food and feed commodities were analyzed for the presence of aflatoxins, fumonisin B1, and zearalenone. Aflatoxins were detected in 40% of the samples analyzed. The concentration of total aflatoxins ranged from 0.1 to 64 microg/kg. The mean concentration ranged from 0.3 microg/kg in sorghum to 23 microg/kg in peanut butter. Peanut butter samples were the most contaminated (71%). No aflatoxins were detected in maize. Fumonisin B1 was detected in 36% of the samples. Maize samples were the most contaminated (85% of the samples) with the concentration ranging from 20 to 1,270 microg/kg. No fumonisin B1 was detected in peanuts, phane, and beans. Zearalenone was only found in 2.6% of the samples analyzed at 40 microg/kg. Aflatoxins were the most common toxins detected in foods and feeds in Botswana. However, fumonisin B1 was more prevalent in maize than aflatoxins or zearalenone.

Aflatoxins↗

Restriction fragment length polymorphism assessment of the heterogeneous nature of maize population GT-MAS:gk and field evaluation of resistance to aflatoxin production by Aspergillus flavus.

Aflatoxin, produced by Aspergillus flavus, is one of the most toxic and carcinogenic substances known and contaminates many agricultural commodities such as corn, peanuts, cottonseed, and tree nuts. The challenge to breeders/plant pathologists is to identify lines that have resistance to aflatoxin production. Maize population GT-MAS:gk has been identified and released as a germplasm with resistance to aflatoxin contamination. In the present study, we assessed genetic divergence in the GT-MAS:gk population using restriction fragment length polymorphism (RFLP) DNA markers to survey 11 selfed inbred lines and conducted field evaluations for the dissimilarities in aflatoxin production among these inbred lines in comparison with a sister population, GT-MAS:pw.nf. The 11 selfed inbred lines were assayed for DNA polymorphism using 113 RFLP markers in 10 linkage groups covering 1,518.2 centimorgans (cM; unit of gene or chromosome size). Considerable variation among the inbreds was detected with RFLP markers, of which 42 probe-enzyme combinations gave 102 polymorphic bands. Cluster analysis based on genetic similarities revealed associations and variations among the tested lines. Three polymorphic groups were distinguished by cluster analysis. Two years of field evaluation data showed that aflatoxin concentrations among the lines were significantly different in both years (P < 0.001). Maturity data were also different. Thus, this study demonstrates that the maize population GT-MAS:gk is heterogeneous and that individuals may be different in resistance to A. flavus infection and aflatoxin production. Therefore, the most resistant lines should be inbred to increase homogeneity, and resistance should be confirmed through progeny testing.

Aflatoxins↗

Determination of aflatoxin B1 in food and feedstuffs in Cuba (1990 through 1996) using an immunoenzymatic reagent kit (Aflacen).

The presence of aflatoxin B1 was analyzed in imported food and feedstuffs of national production in the period of 1990 through 1996, destined to animal and human consumption using an immunoenzymatic reagent kit (Aflacen, Ckure, la Habana, Cuba) with a detection limit of 0.3 microg/kg. It was found that the 17.04% of a total of 4,594 analyzed samples presented aflatoxin B1, and the biggest percentages were in sorghum and peanut with an 83.3 and 40.4%, respectively. The corn, oat, wheat, and soy are fundamental raw ingredients in the elaboration of concentrates. Percentages of contamination with aflatoxin B1 of 23.3, 10.7, 25, and 4.6 were found in corn, oat, wheat, and soy, respectively. Other analyzed foods like rice, beans, and peas presented percentages of contamination with aflatoxin B1 inferior to 5% of the analyzed samples. It was found that more than 455 samples surpassed the value of 10 microg/kg. Corn and peanut products present a high demand in population showing levels of contamination superior to 50 microg/kg. The 11.3% of the samples contaminated with aflatoxin B1 have values between 1 and 20 microg/kg, where peanut and concentrates show the highest percentages (21.9 and 18.7), respectively. These results show levels of aflatoxin B1 in the population that constitute a great risk for human and animal health.

Aflatoxin B1↗

Aflatoxin production in peanut lines selected to represent a range of linoleic acid concentrations.

To determine whether concentrations of linoleate in peanut (Arachis hypogaea L.) seed oil could be used to predict an ability to support aflatoxin production, seeds of genotypes representing a range of linoleate content were inoculated with Aspergillus flavus Link ex Fries and assayed for aflatoxin content. Seeds were blanched and quartered, inoculated with conidia of A. flavus, placed on moistened filter paper in petri dishes, and incubated for 8 days at 28 degrees C. Multiple regression analysis was used to account for the variation among lines with the use of fatty acid concentrations as independent variables. In test 1, linoleate accounted for 39 to 44% of the variation among lines for aflatoxin B1 and B2 and total aflatoxin (26 to 27% after log transformation). Oleate accounted for substantial additional variation (27 to 29%) among lines (20 to 23% after log transformation). Other fatty acids accounted for small fractions of among-line variation. In test 2, linoleate accounted for about 35 to 44% of the variation among entries across traits (29 to 37% for log-transformed data); arachidate accounted for 19 to 29% (27 to 33% after log transformation). Eicosenoate accounted for a small part of the total entry variation. In both experiments, residual variation among entries was significant. Low-linoleate lines consistently contained more aflatoxin, whereas normal- to high-linoleate lines contained variable amounts. Although fatty acid concentrations accounted for significant portions of genetic variation, it is not practical to use them as predictors for susceptibility to aflatoxin contamination, especially for lines in the normal range for oleate and linoleate.

Aflatoxins↗

Aflatoxin exposure and DNA damage in the comet assay in individuals from the Gambia, West Africa.

The single cell gel electrophoresis assay (Comet assay) was used to measure DNA damage in peripheral lymphocytes from a group of individuals from The Gambia in order to determine whether such damage could be associated with increased exposure to aflatoxin in this population. Responses obtained were correlated to responses previously obtained [1] in a cross-sectional study in the same individuals of various cytogenetic alterations [chromosomal aberrations, micronuclei (crest positive and negative staining), and sister chromatid exchanges], and aflatoxin-albumin adducts. Analysis of variance methods were used to assess the effects of smoking, GSTM1 genotype, sex, age, and smoking status. A comparison was also made between The Gambian individuals and a group of healthy, non-smoking volunteers in the United Kingdom where aflatoxin exposure would be expected to be low. From the earlier study [1], it was determined that the levels of the sister chromatid exchanges and micronuclei were higher in The Gambian group than in a European group where aflatoxin exposure was lower, but that there were no correlations between the adduct levels and the cytogenetic abnormalities at the individual level. In the present study, DNA damage as measured in the Comet assay was not significantly higher than in the healthy United Kingdom volunteers. In addition, there were no associations between cytogenetic damage, GSTM1 genotype, age, sex, lifestyle factors (smoking and aflatoxin exposure), and Comet response at the individual level. Comet response was higher in females than males in The Gambia if one outlier was excluded from analysis and not taking into account other sources of variability. It would appear that DNA damage as measured in the Comet assay in peripheral blood lymphocytes is not a sensitive genotoxic marker of aflatoxin exposure in this population.

Adolescent↗

The effect on performance and biochemical parameters when soil was added to aflatoxin-contaminated poultry rations.

The effects of silty clay loam soil on the performance and biochemical parameters of chicks were investigated when the soil was added to aflatoxin B1 (AFB1)-contaminated diets. One hundred 14-d-old White Leghorn chicks were fed a control ration (clean corn), a low aflatoxin-contaminated ration (120 ng AFB1/g), a high aflatoxin-contaminated ration (700 ng AFB1/g), or high aflatoxin-contaminated rations (700 ng AFB1/g) +10% or 25% soil. Body weight, feed consumption and blood samples were monitored weekly. Decreased feed consumption, body weight gain and efficiency of feed utilization, increased SGOT and LDH activities, and cholesterol and triglyceride concentrations, and decreased uric acid concentrations and ALP activity were observed in the chicks fed the high aflatoxin-contaminated ration without soil. Hepatomegaly was prominent in chicks fed the high aflatoxin-contaminated ration without soil, and some livers had extensive hepatocyte vacuolation, hepatocellular swelling, fatty change and hydropic degeneration, and stained positive for fat accumulation. Addition of soil reduced the detrimental effects of AFB1 for some parameters, although the reduction was less when 10% soil was fed compared with the 25% soil feeding.

Aflatoxin B1↗

Comparative mutagenicity of palmotoxin Bo and aflatoxins B1 and M1.

The mutagenicity of palmotoxin Bo and of aflatoxin M1 relative to that of aflatoxin B1, the potent mutagen, was studied in five Ames' Tester Strains of Salmonella typhimurium (TA-98, TA-100, TA-1535, TA-1537, TA-1538). Aflatoxins B1 and M1 are both highly mutagenic in a microsome-mediated system in TA-100. The prediction of the relative carcinogenicity of aflatoxin M1 to aflatoxin B1 posed by the mutation of TA-100 is probably more authentic than TA-87. The mutagenic potency of palmotoxin Bo is less than that of either aflatoxin B1 or M1.

Aflatoxins↗

Testing shelled corn for aflatoxin, Part I: estimation of variance components.

The variability associated with testing lots of shelled corn for aflatoxin was investigated. Eighteen lots of shelled corn were tested for aflatoxin contamination. The total variance associated with testing shelled corn was estimated and partitioned into sampling, sample preparation, and analytical variances. All variances increased as aflatoxin concentration increased. With the use of regression analysis, mathematical expressions were developed to model the relationship between aflatoxin concentration and the total, sampling, sample preparation, and analytical variances. The expressions for these relationships were used to estimate the variance for any sample size, subsample size, and number of analyses for a specific aflatoxin concentration. Test results on a lot with 20 parts per billion aflatoxin using a 1.13 kg sample, a Romer mill, 50 g subsamples, and liquid chromatographic analysis showed that the total, sampling, sample preparation, and analytical variances were 274.9 (CV = 82.9%), 214.0 (CV = 73.1 %), 56.3 (CV = 37.5%), and 4.6 (CV = 10.7%), respectively. The percentage of the total variance for sampling, sample preparation, and analytical was 77.8, 20.5, and 1.7, respectively.

Aflatoxins↗

Simultaneous quantitation of Aspergillus flavus/A. parasiticus and aflatoxins in peanuts.

A method was developed for simultaneous quantitation of Aspergillus flavus/A. parasiticus and aflatoxins in peanuts. Peanut samples were ground with an equal weight of water in a vertical cutter mixer to produce a slurry. Separate subsamples were taken for dilution-plating to determine total colony forming units (CFU)/g of A. flavus/A. parasiticus and for liquid chromatographic analysis to determine aflatoxin concentrations. Dry-grinding peanuts for homogenization of aflatoxins produced high temperatures that killed most of the A. flavus/A. parasiticus propagules. Addition of water to produce a slurry kept the temperature from rising above levels that killed the fungi. A 7 min grind time provided optimal homogenization for both the fungi and aflatoxins, so long as the temperature of the slurry did not exceed 45 degrees C. In the analysis of 60 shelled peanut samples, total aflatoxin concentrations ranged from 0 to 10,000 ng/g and total A. flavus/A. parasiticus ranged from 1.4 x 10(3) to 3.2 x 10(6) CFU/g. Regression analysis showed a significant positive correlation (p < 0.0001) between the quantities of A. flavus/A. parasiticus and aflatoxin (R2 = 0.82).

Aflatoxins↗

[Specific features of albumin interactions with hemiacetals of aflatoxins and sterigmatocystin].

Aflatoxin B2a (AB2a), aflatoxin G2a (AG2a), and hemiacetal of sterigmatocystin have been shown to form immunoreactive conjugates with albumin. The conjugates were formed following incubation of solution mixtures at room temperature for 1 h, as demonstrated by spectrophotometry and enzyme immunoassay. Anti-AB2a antibodies reacted with AB2a, aflatoxin B1, and aflatoxin AB2 (100, 8.8, and 5.9%, respectively); a similar result was obtained for anti-AG2a antibodies reacting with AG2a, aflatoxin G1, and aflatoxin AG@2 (100, 2.5, and < 1.0%, respectively). Binding of anti-AB2a and anti-AG2a antibodies to solid-phase conjugates of AB2a or AG2a exhibited similar analytical characteristics.

Aflatoxins↗

[Studies on chemical analysis of mycotoxin (XXI). A rapid analytical method for aflatoxins by immunoaffinity column chromatography and high performance liquid chromatography].

An analytical method for aflatoxins using solvent extraction, affinity column (containing monoclonal antibodies against aflatoxins bound onto agarose) clean-up and high performance liquid chromatographic determination was applied to the analysis of aflatoxins in peanuts, pistachio nuts, white and black peppers and corn. Recoveries of aflatoxins B1, B2, G1 and G2 spiked to peanuts, pistachio nuts and corn at the level of 20 ppb were 77.4-81.4, 70.1-77.9 and 72.3-94.8%, respectively. In the case of white and black peppers, because the spiked aflatoxin recovery rate was low, further purification steps should be studied. The stability of the affinity column was good. This procedure is recommended as a rapid and reliable method for the analysis of aflatoxins in the three kinds of foods, peanuts, pistachio nuts and corn.

Aflatoxins↗

Studies of aflatoxins in Chiang Mai, Thailand.

Aflatoxins become an economical problem in our country due to its contamination in agricultural commodities for export. The toxin may also cause hepatoma and liver diseases in the Thais as well. It is, therefore, necessary to search for and to develop efficient technology to combat and control such dangerous mold and mycotoxins. This paper is a collection of our previous and present studies towards reduction of risk from aflatoxins in foods and feedstuffs. The investigation of mold and aflatoxin contamination in local foods and feedstuffs in Chiang Mai area was made. The inhibitory effect of garlic extract on growth of Aspergillus flavus and its aflatoxin production was demonstrated. Detoxification of aflatoxins by chemicals such as ammonium carbonate, ammonium bicarbonate and ammonium benzoate was also shown. Preventions of toxigenic mold growth and its aflatoxin production by means of some food preservative were reported. Modifications of such effective chemicals were investigated for safety in future application.

Aflatoxins↗

Immunoaffinity column cleanup with liquid chromatography using post-column bromination for determination of aflatoxin B1 in cattle feed: collaborative study.

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 contaminated with aflatoxin B1, 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 contaminated 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.

Aflatoxin B1↗

The role of aflatoxin-contaminated food materials and HCV in developing hepatocellular carcinoma in Al-Sharkia Governorate, Egypt.

Aflatoxins, particularly aflatoxin B1 (AFB1) have been recognized as one of the most potent chemical carcinogen. In Egypt, HCV is prevalent. The progressive nature of HCV-related liver diseases was found to be influenced by other factors. In this paper, the role of aflatoxin contamination in the onset of liver cancer in HCV-infected patients was studied. The quantitative identification of the possible aflatoxins contamination in six urban and eleven rural areas using high performance liquid chromatography technique, revealed that corn, wheat, pea nut, lupine "termis", white rice, cowpea "lobiya", fava bean and brown rice showed the prevalence of AFB1 to be 64.7%, 53%, 53%, 47%, 47%, 41%, 29.4% & 29.4% respectively. A positive correlation was found between aflatoxin and positive HCV-PCR together with liver disease progression to G3S3, the indicative of hepatocellular carcinoma. Such correlation was not fully understood, but the oncogene amplification caused by HCV-infection may be aggravated by the consumption of aflatoxin contaminated raw food materials or their products.

Aflatoxins↗