An isolate of Fusarium heterosporum Nees ex Fr. that parasitizes the honeydew and immature sclerotia of Claviceps paspali was shown to be highly toxigenic to day-old chickens. The major toxicity of culture extracts of the fungus was due to the presence of several trichothecene mycotoxins. Six trichothecenes were isolated and chemically identified by spectroscopic methods as T-2 toxin (1), HT-2 toxin (2), T-2 tetraol (6), 3alpha,4beta,dihydroxy-15-acetoxy-8alpha-(3-hydroxy-3-methylbutyryloxy)-12,13-epoxytrichothec-9-ene (3), 3alpha,4beta,15-trihydroxy-8alpha-(3-hydroxy-3-methylbutyryloxy)-12,13-epoxytri chothec-9-ene (5), and 3alpha,4beta,8alpha-trihydroxy-15-acetoxy-12,13-epoxytrichothec-9-ene (4). The acute toxicities of the trichothecenes were compared to the effects of T-2 toxin in day-old chickens and on the back skins of rabbits. The comparative inhibitory effects of the trichothecene mycotoxins in the Triticum coleoptile bioassays were also determined.

Animals↗

PubMedDate not supplied

Comparison of two ELISA screening tests with liquid chromatography for determination of aflatoxins in raw peanuts.

A study was conducted to evaluate the performance of 2 enzyme-linked immunosorbent assays (ELISA) for rapidly screening samples of peanuts for the presence of aflatoxin. The EZ-Screen Quick Card Test and the Afla-10 Cup Test were compared with liquid chromatography in duplicate analyses of common extracts of peanuts contaminated in the range of 0-70 ppb (ng/g). Each assay properly identified 95% of samples containing no detectable aflatoxin as negative and greater than 97% of samples containing greater than 10 ppb aflatoxin as positive. The card test, which had a 20 ppb detection threshold, identified as positive 32 of 34 samples in the 11-20 ppb range. This indicates that the card test might actually have a detection threshold closer to 10 ppb. Most of the errors associated with the assays occurred on samples containing less than 10 ppb aflatoxin. The cup and card tests identified 76 and 67% of the samples, respectively, as negative, in the range of 4-10 ppb. For samples either negative or contaminated above their detection thresholds for the assays, the methods are well suited for use as rapid screening tests.

Aflatoxins↗

PubMedDate not supplied

Interaction of aflatoxin B1 and cyclopiazonic acid toxicities.

Toxic properties of the mycotoxins cyclopiazonic acid and aflatoxin B1 have been analyzed separately and in combination by monitoring their effects on luminescence in the marine bacterium Photobacterium phosphoreum, Strain NCMB 844. Genotoxicity was analyzed with a dark mutant of this organism whose reversion to the bioluminescent condition is stimulated by compounds attacking guanine sites in deoxyribonucleic acids. In this assay, cyclopiazonic acid, unlike aflatoxin B1, is not enhanced by cyclopiazonic acid when the two mycotoxins are assayed in combination. Cytotoxicity was assessed by the diminution of bioluminescence in a separate assay system with strain NRRLB-1177 of P. phosphoreum. Cyclopiazonic acid is more cytotoxic than aflatoxin B1, and concentrations of cyclopiazonic acid required for cytotoxicity decreases with time, whereas aflatoxin B1 cytotoxic expression does not change significantly with time under most assay conditions. Aflatoxin B1 and cyclopiazonic acid assayed as a dose pair indicate that these mycotoxins elicit their effects by independent modes of action.

Aflatoxin B1↗

PubMedDate not supplied

Rapid determination of aflatoxins in raw peanuts by liquid chromatography with postcolumn iodination and modified minicolumn cleanup.

A method is described for rapid cleanup followed by reverse-phase liquid chromatographic (LC) quantitation of aflatoxins in raw peanuts. A modified minicolumn cleanup is used for sample preparation, and a preliminary estimation of aflatoxin content by minicolumn can be made so that highly contaminated samples can be diluted before LC analysis. The use of the simple, quick minicolumn cleanup eliminates the need for further column or cartridge cleanup, thus greatly reducing sample preparation time. Sensitive quantitation is achieved using a phenyl column, a mobile phase of water-tetrahydrofuran (80 + 20, v/v), and postcolumn derivatization with water-saturated iodine followed by fluorescence detection. The recoveries of aflatoxins B1, B2, G1, and G2 from peanut meal spiked at 3 levels ranged from 71.7 to 88.3% (average 80%) with coefficients of variation from 2.7 to 10.4%.

Aflatoxins↗

PubMedDate not supplied

Liquid chromatographic determination of cyclopiazonic acid in poultry meat.

A liquid chromatographic procedure has been developed for the determination of cyclopiazonic acid (CPA) in poultry meat. CPA is extracted from ground meat with chloroform-methanol (80 + 20), partitioned into 0.1N sodium hydroxide, acidified, and extracted into dichloromethane. An interfering component of meat is removed by transferring the dichloromethane extract to a minicolumn containing silica gel and washing the column with petroleum ether and chloroform. CPA is eluted with methanol-acetic acid (99 + 1), and subjected to ligand-exchange liquid chromatography. Recovery of CPA from 40 separate samples of meat spiked with CPA at levels from 0.016 to 15.6 ppm was 70.4 +/- 14.1%. Analysis of meat from a chicken orally dosed with 10 mg CPA/kg body weight revealed that 14.5% of the dose was in muscle 48 h after administration.

Animals↗

PubMedDate not supplied

Extraction of aflatoxins from naturally contaminated peanuts with different solvents and solvent/peanut ratios.

A study was conducted to compare the extraction efficiencies of aflatoxins (B1, B2, G1, and G2) from low, medium, and high concentrations (approximately 10, 100, and 1000 ppb, respectively) of naturally contaminated peanut samples by using different solvents and solvent-to-peanut sample ratios. Solvents used were 80:20 methanol/water at 2:1, 3:1, and 5:1 solvent to sample ratios; 60:40 methanol/water at 5:1 solvent to sample ratio; and 90:10 acetonitrile/water at 2:1 and 4:1 solvent to sample ratios. The solvents 80:20 methanol/water at a 3:1 ratio of solvent to sample and 90:10 acetonitrile/water at a 2:1 ratio were consistently more efficient than all other solvents, regardless of the aflatoxin concentration. The solvent 90:10 acetonitrile/water at a 4:1 solvent-to-sample ratio was consistently the least efficient regardless of aflatoxin concentration based on analytical results with liquid chromatography (LC). The results are relevant in selecting an appropriate extraction solvent for use at peanut grading points.

Aflatoxins↗

PubMedDate not supplied

Variability among peanut subsamples prepared for aflatoxin analysis with four mills.

The variability in aflatoxin concentration among peanut subsamples ground with 4 different mills was evaluated. Twenty 2 kg samples of naturally contaminated peanuts were ground in a Dickens subsampling mill (DM), a Stephan model UM-12 vertical cutter mixer (SM), and a Robot Coupe model RS16Y-1 vertical cutter mixer (RC1). Twenty 4 kg samples were ground in the DM, SM, and a Robot Coupe model R10P vertical cutter mixer (RC2). From each 2 kg sample, ten 100 g subsamples were withdrawn, and from each 4 kg samples, ten 200 g subsamples were withdrawn. Subsamples were analyzed for aflatoxin by liquid chromatography. The coefficient of variation (CV) among each set of 10 subsamples was determined for each sample, and the CVs for each sample size were ranked and analyzed by the Kruskal-Wallis test of ranks. For 2 kg samples, the CVs for the samples ground in RC1 ranked significantly lower than those for samples ground in DM and SM. For 4 kg samples, the CVs for samples ground in RC2 and SM were significantly lower than that for samples ground in DM. The averages of the CVs for 2 kg samples were 17.2% (RC1), 32.8% (SM), and 40.6% (DM). The averages of the CVs for 4 kg samples were 21.2% (RC2), and 26.0% (SM), and 47.0% (DM).

Aflatoxins↗

PubMedDate not supplied

Performance of two immunochemical assays in the analysis of peanuts for aflatoxin at 37 field laboratories.

A study was conducted to measure the precision of 2 rapid aflatoxin assay systems in use at 37 peanut buying points during the 1991 harvest season. Aflatoxin laboratories were established at the 37 buying points to analyze peanut samples from all incoming farmers' stock loads as part of a joint project sponsored by various segments of the U.S. peanut industry and the U.S. Department of Agriculture. Eighteen laboratories were equipped with Neogen's veratox FSP rapid assay system, whereas 19 laboratories used Vicam's Aflatest rapid assay system. To monitor the performance of the field laboratories during the project, 3 portions of each of six 27 kg samples of ground peanuts were sent to each laboratory for analysis over a period of 6 weeks. Aflatoxin concentrations ranged from 0 to 300 ng/g when eight 200 g subsamples of each sample were analyzed by liquid chromatography (LC). For the 5 samples contaminated with aflatoxin, relative standard deviations for repeatability (RSDr) for laboratories using veratox FSP ranged from 18.66 to 53.29%, and the relative standard deviations for reproducibility (RSDR) ranged from 22.79 to 59.29%. For laboratories using the Aflatest system, RSDr values ranged from 18.70 to 41.48%, and RSDR values ranged from 23.84 to 47.56%. Horwitz ratios < 2.0 were found for 4 of the 5 contaminated samples for both methods, indicating that the overall precision of the 2 methods used in the project was good. Mean aflatoxin concentrations, as determined with the rapid assay systems, were generally lower than those determined by LC, particularly for more highly contaminated samples.(ABSTRACT TRUNCATED AT 250 WORDS)

Aflatoxins↗

PubMedDate not supplied

Estimating aflatoxin in farmers' stock peanut lots by measuring aflatoxin in various peanut-grade components.

Five, 2 kg test samples were taken from each of 120 farmers' stock peanut lots contaminated with aflatoxin. Kernels from each 2 kg sample were divided into the following grade components: sound mature kernels plus sound splits (SMKSS), other kernels (OK), loose shelled kernels (LSK), and damaged kernels (DAM). Kernel mass, aflatoxin mass, and aflatoxin concentration were measured for each of the 2400 component samples. For 120 lots tested, average aflatoxin concentrations in SMKSS, OK, LSK, and DAM components were 235, 2543, 11,775, and 69,775 ng/g, respectively. Aflatoxins in SMKSS, OK, LSK, and DAM components represented 6.9, 7.9, 33.3, and 51.9% of the total aflatoxin mass, respectively. Cumulatively, 3 aflatoxin risk components--OK, LSK, and DAM--accounted for 93.1% of total aflatoxin, but only 18.4% percent of test sample mass. Correlation analysis suggests that the most accurate predictor of aflatoxin concentration in the lot is the cumulative aflatoxin mass in the high 3 risk components OK + LSK + DAM (correlation coefficient, r = 0.996). If the aflatoxin in the combined OK + LSK + DAM components is expressed in concentration units, r decreases to 0.939. Linear regression equations relating aflatoxin in OK + LSK + DAM to aflatoxin concentration in the lot were developed. The cumulative aflatoxin in the OK + LSK + DAM components was not an accurate predictor (r = 0.539) of aflatoxin in the SMKSS component. Statistical analyses of 3 other data sets published previously yielded similar results.

Aflatoxins↗

PubMedDate not supplied

Immunoaffinity column as cleanup tool for a direct competitive enzyme-linked immunosorbent assay of cyclopiazonic acid in corn, peanuts, and mixed feed.

An immunoaffinity column (IAC) for cyclopiazonic acid (CPA) was prepared by coupling a CPA-specific monoclonal antibody to CNBr-activated sepharose 4B. A direct competitive enzyme-linked immunosorbent assay (dc-ELISA) was used to study the chromatographic behavior of a 0.2 mL gel column with a binding capacity of 4 micrograms CPA/column as well as to evaluate its efficacy as a cleanup tool for analysis of naturally occurring CPA. Sample extract either in buffer solution or in a solution containing up to 35% methanol could be loaded onto the column. After the column is washed with 5 mL deionized water and 5 mL 50% methanol, CPA could be quantitatively eluted with 2 mL 100% methanol. The column could be regenerated at least 10 times by washing with 10 mL equilibrating buffer and then storing in a cold room overnight before reuse. Recoveries of CPA added to corn, peanut, and mixed feed extracts in the range 10-200 ng/g were 88-105, 86-100, and 90-110%, respectively. Detection limits were 2.0, 4.4, and 4.7 ng/g for corn, mixed feed, and peanuts, respectively. Twenty-two peanut samples naturally contaminated with CPA were subjected to both IAC and solvent partition cleanup followed by dc-ELISA. Although a good correlation between data obtained from IAC-dc-ELISA and from SP-dc-ELISA (r = 0.75, p < 0.0001) was obtained, the slope of the linear regression was low (0.67), indicating loss during solvent partition cleanup. The overall data showed that the combination of IAC and dc-ELISA is an effective method for CPA analysis.

Animal Feed↗