Hamed K. Abbas

Corn (maize, Zea mays) is susceptible to contamination with aflatoxins, fumonisins and other mycotoxins, particularly in the southeastern USA. In principle, mycotoxin contamination could be reduced in commercial corn hybrids with shorter growing seasons by planting at dates which minimize plant stress during the critical kernel-filling period. To evaluate this strategy, commercial Bt and non-Bt hybrids were planted in Arkansas in mid-April and early May of 2002, 2004 and 2005. The mid-April planting date resulted in lower aflatoxin contamination in harvested corn each yr and in significantly less frequent contamination above a regulatory action level in 2005 and overall than did the early-May planting date in both Bt and non-Bt corn. The mid-April planting date resulted in significantly lower total fumonisin contamination in harvested corn and in less frequent contamination above a regulatory advisory level than the early May planting date in 2 of 3 yr and overall in both Bt and non-Bt corn. All fumonisin subtypes studied were reduced. Frequent co-occurrence of aflatoxin and fumonisin was observed. Fumonisin levels averaged lower in Bt hybrids than in non-Bt hybrids at all plantings. Reduced aflatoxin and fumonisin contamination with mid-April planting could not be explained by any measure of heat stress during the kernel-filling period.

The ability of two non-aflatoxigenic Aspergillus flavus Link isolates (CT3 and K49) to reduce aflatoxin contamination of corn was assessed in a 4-year field study (2001–2004). Soil was treated with six wheat inoculant treatments: aflatoxigenic isolate F3W4; two non-aflatoxigenic isolates (CT3 and K49); two mixtures of CT3 or K49 with F3W4; and an autoclaved wheat control, applied at 20 kg ha⁻¹. In 2001, inoculation with the aflatoxigenic isolate increased corn grain aflatoxin levels by 188% compared to the non-inoculated control, while CT3 and K49 inoculation reduced aflatoxin levels in corn grain by 86 and 60%, respectively. In 2002, the non-toxigenic CT3 and K49 reduced aflatoxin levels by 61 and 76% compared to non-inoculated controls, respectively. In 2001, mixtures of aflatoxigenic and non-aflatoxigenic isolates had little effect on aflatoxin levels, but in 2002, inoculation with mixtures of K49 and CT3 reduced aflatoxin levels 68 and 37% compared to non-inoculated controls, respectively. In 2003 and 2004, a low level of natural aflatoxin contamination was observed (8 ng g⁻¹). However, inoculation with mixtures of K49 + F3W4 and CT3 + F3W4, reduced levels of aflatoxin 65–94% compared to the aflatoxigenic strain alone. Compared to the non-sclerotia producing CT3, strain K49 produces large sclerotia, has more rapid in vitro radial growth, and a greater ability to colonize corn when artificially inoculated, perhaps indicating greater ecological competence. Results indicate that non-aflatoxigenic, indigenous A. flavus isolates, such as strain K49, have potential use for biocontrol of aflatoxin contamination in southern US corn.

Multiple seeded cocklebur has been found in the last decade in Texas, and described as a biotype of Xanthium strumarium L with up to 25 seeds per bur instead of the usual two. The multiple seeded bur typically produces up to nine seedlings, causing concern that it may be harder to control than normal seeded common cocklebur. The efficacies of a series of fungal and conventional commercial herbicides have been compared in the greenhouse on seedlings of multiple seeded cocklebur from Texas (MSC‐TX) and normal common cockleburs from Texas (NCC‐TX), Arkansas (NCC‐AR), Illinois (NCC‐ILL) and two from Mississippi (NCC‐MS#1, NCC‐MS#2). Three measures of herbicidal activity (reductions in plant height and dry weight, and mortality) were used. The fungal herbicide Alternaria helianthi (Hansf) Tubaki & Nishihara at 1 × 10⁵ conidia ml⁻¹ + 2 g liter⁻¹ Silwet L‐77 with an 8‐h dew period was an effective herbicide with all biotypes, as were the commercial chemical herbicides chlorimuron (14.8 g ha⁻¹), imazaquin (29.6 g ha⁻¹), sodium hydrogen methylarsonate (MSMA; 279.1 g ha⁻¹) and imazethapyr (39.5 g ha⁻¹). The membrane‐disrupting organic arsenical MSMA was effective with all biotypes, whereas commercial chemical herbicides which act by inhibiting branched‐chain amino acid synthesis (chlorimuron, imazaquin and imazethapyr) were less effective against normal seeded common cocklebur biotypes with short stature. These studies showed that multiple seeded cocklebur was at least as susceptible to the biological agent A helianthi and to the conventional commercial herbicides studied as were normal seeded cockleburs, suggesting that existing methods should be adequate to control this novel biotype. Copyright © 2005 Society of Chemical Industry

This study compared cultural and analytical methods for detecting aflatoxin production by Aspergillus species. Aspergillus isolates were obtained from various Mississippi Delta crops (corn, peanut, rice, cotton) and soils. Most of the isolates (99%) were A. flavus and the remainder comprised A. parasiticus and A. nomius. The following three cultural methods were evaluated on potato dextrose agar: fluorescence (FL) on β-cyclodextrin-containing media (CD), yellow pigment (YP) formation in mycelium and medium, and color change after ammonium hydroxide vapor exposure (AV). Aflatoxins in culture extracts were confirmed by thin-layer chromatography (TLC) and quantified by enzyme-linked immunosorbent assay (ELISA). Of the 517 isolates, 314 produced greater than 20 ng/g of total aflatoxin based on ELISA, and 180 produced greater than 10 000 ng/g of aflatoxin in the medium. Almost all the toxigenic isolates (97%) were confirmed by TLC as producers. Of the toxigenic isolates, as determined by ELISA, 93%, 73%, and 70% gave positive FL, YP, and AV responses, respectively. Of the 203 isolates producing less than 20 ng/g of aflatoxin, 20%, 6%, and 0% of respective FL, YP, and AV methods gave false-positive responses. The 9% false-positive results from TLC fall within this range. This study showed good agreement among all tested cultural methods. However, these cultural techniques did not detect aflatoxin in all cultures that were found to produce aflatoxins by ELISA, LC/MS, and TLC. The best results were obtained when the AV color change and CD fluorescence methods were used together, yielding an overall success rate comparable to TLC but without the need for chemical extraction and the time and expense of TLC.Key words: aflatoxins, analytical methods, Aspergillus flavus, Aspergillus parasiticus, cultural methods, β-cyclodextrin, fluorescence enhancers, mycotoxins, yellow pigment.

The phytotoxicity and mammalian cytotoxicity of four nontrichothecene mycotoxins (apicidin, sambutoxin, wortmannin, HC-toxin) were compared. Phytotoxicity was evaluated in terms of electrolyte leakage, growth inhibition, and reduction in chlorophyll content. Based on the parameters evaluated, the relative order of phytotoxicity to duckweed (Lemna pausicostata L.) was wortmannin > HC-toxin > apicidin >> sambutoxin. A 48-hr exposure to 10 microM wortmannin, HC-toxin or apicidin caused electrolyte leakage from duckweed. The IC50 values for growth inhibition and chlorophyll reduction for wortmannin, HC-toxin, and apicidin were 0.2 and 2.6 microM, 15.4 and 12.6 microM, and 27.7 and 45.3 microM, respectively. Based on the parameters measured, a 72-hr exposure to 100 microM sambutoxin was not toxic to duckweed. Kudzu (Pueraria lobata L.) leaf disc assays revealed a similar trend in relative toxicities, but higher mycotoxin concentrations were required to elicit phytotoxic effects compared to duckweed. All four mycotoxins were cytotoxic to four mammalian cell cultures. However, in contrast to plants, wortmannin was the least toxic (IC50 = 10 to 20 microM) and sambutoxin exhibited a high level of toxicity (IC50 = 0.5 to 1 microM).

Corn samples suspected of causing refusal-to-eat syndrome in dairy cattle were examined mycologically. Fusarium moniliforme (14 isolates) and F. proliferatum (12 isolates) were the predominant fungi present. These isolates were tested for mycotoxin production on rice at 25 degrees C. Each strain of F. moniliforme produced fumonisin B1 (FB1: 378-15,600 ppm) and fumonisin B2 (FB2: 2-1050 ppm). Each strain of F. proliferatum produced moniliformin (45-16,000 ppm), FB1 (27-6140 ppm), and FB2 (5-1550 ppm). In addition, a new Fusarium metabolite of molecular composition C21H38N2O6 was produced by 10 of the F. moniliforme isolates and 7 of the F. proliferatum isolates. The metabolite's 1H- and 13C-NMR, HRFAB/MS and IR spectra indicate an alpha amino acid. It is toxic to Lemna minor L. duckweed (LD50 100 micrograms/mL).