Biological Control of Aflatoxin Contamination in U.S. Crops and the Use of Bioplastic Formulations of Aspergillus flavus Biocontrol Strains To Optimize Application Strategies

Aflatoxin contamination has a major economic impact on crop production in the southern United States. Reduction of aflatoxin contamination in harvested crops has been achieved by applying nonaflatoxigenic biocontrol Aspergillus flavus strains that can out-compete wild aflatoxigenic A. flavus, reducing their numbers at the site of application. Currently, the standard method for applying biocontrol A. flavus strains to soil is using a nutrient-supplying carrier (e.g., pearled barley for Afla-Guard). Granules of Bioplastic (partially acetylated corn starch) have been investigated as an alternative nutritive carrier for biocontrol agents. Bioplastic granules have also been used to prepare a sprayable biocontrol formulation that gives effective reduction of aflatoxin contamination in harvested corn kernels with application of much smaller amounts to leaves later in the growing season. The ultimate goal of biocontrol research is to produce biocontrol systems that can be applied to crops only when long-range weather forecasting indicates they will be needed

Effect of temperature, rainfall and planting date on aflatoxin and fumonisin contamination in commercial Bt and non-Bt corn hybrids in Arkansas

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.Le maïs (Zea mays) est sensible à la contamination par les aflatoxines, les fumonisines et d'autres mycotoxines, surtout dans le sud-est des États-Unis. En principe, la contamination par les mycotoxines pourrait être diminuée chez les hybrides commerciaux de maïs par des saisons de croissance plus courtes en plantant à des dates qui minimisent le stress sur les plantes au moment de la période critique du remplissage des grains. Pour évaluer cette stratégie, des hybrides commerciaux Bt et non Bt ont été semés en Arkansas de la mi-avril au début de mai 2002, 2004 et 2005. Pour toutes les années, tant pour le maïs Bt que le non Bt, le grain issu des semis de la mi-avril était moins contaminé aux aflatoxines que celui semé au début de mai. De même, la contamination supérieure aux niveaux légalement acceptés a été moindre en 2005 et dans l'ensemble pour les semis de la mi-avril. Avec les semis de la mi-avril, il y avait significativement moins de contamination par les fumonisines dans le grain récolté et moins de contamination supérieure aux niveaux légalement acceptés qu'avec les semis du début de mai pour deux des trois années et dans l'ensemble, tant pour le maïs Bt que celui non Bt. La quantité de tous les sous-types de fumonisines étudiés a été diminuée. La présence simultanée d'aflatoxines et de fumonisines a fréquemment été observée. Pour tous les semis, les quantités de fumonisines des hybrides Bt ont été inférieures en moyenne à celles des hybrides non Bt. Les moindres contaminations par les aflatoxines et les fumonisines avec les semis de la mi-avril n'ont pu être expliquées par aucune des mesures du stress causé par la chaleur lors de la période du remplissage des grains

CHARACTERIZATION STUDIES ON A TETRAHYDROCURCUMIN-ZINC COMPLEX

Objective: Preparation and characterization studies on tetrahydrocurcumin complexed with zinc, with particular reference to the location of zinc in the complex. Methods: Structural characteristics of tetrahydrocurcumin and its complex with zinc were compared using elemental analysis, mass spectrometry (MS), proton, and carbon-13 nuclear magnetic resonance spectroscopy (NMR), ultraviolet-visible (UV) absorption spectroscopy, and Fourier transform infrared (FTIR) spectroscopy. Results: MS data indicates a Zn molecule holds two THCur components together. NMR data provide evidence that the zinc ion is associated with the 1,3-diketone part of the linker region of the associated THCur. FTIR data is consistent with zinc interaction with the enol tautomer of the 1,3-diketone at the center of the linker region of THCur. UV data indicate that a zinc-dependent shift in absorbance maximum is consistent with changes in the structure of THCur resulting from complexation with zinc. Together, this data indicates the complexation of zinc with tetrahydrocurcumin is consistent with zinc linking two molecules of tetrahydrocurcumin together by binding to the enol forms of the 1,3-diketone moieties located in the linker regions between the aromatic rings. Conclusion: The spectral properties of the tetrahydrocurcumin-zinc complex are consistent with a structure in which zinc is encased in two tetrahydrocurcumin moieties. Additional studies are needed to determine if this structure results in altered bioavailability, antioxidant activity and other properties important for pharmaceutical development

Toxin production in soybean (Glycine max L.) plants with charcoal rot disease and by Macrophomina phaseolina, the fungus that causes the disease

Charcoal rot disease, caused by the fungus Macrophomina phaseolina, results in major economic losses in soybean production in southern USA. M. phaseolina has been proposed to use the toxin (-)-botryodiplodin in its root infection mechanism to create a necrotic zone in root tissue through which fungal hyphae can readily enter the plant. The majority (51.4%) of M. phaseolina isolates from plants with charcoal rot disease produced a wide range of (-)-botryodiplodin concentrations in a culture medium (0.14-6.11 μg/mL), 37.8% produced traces below the limit of quantification (0.01 μg/mL), and 10.8% produced no detectable (-)-botryodiplodin. Some culture media with traces or no (-)-botryodiplodin were nevertheless strongly phytotoxic in soybean leaf disc cultures, consistent with the production of another unidentified toxin(s). Widely ranging (-)-botryodiplodin levels (traces to 3.14 μg/g) were also observed in the roots, but not in the aerial parts, of soybean plants naturally infected with charcoal rot disease. This is the first report of (-)-botryodiplodin in plant tissues naturally infected with charcoal rot disease. No phaseolinone was detected in M. phaseolina culture media or naturally infected soybean tissues. These results are consistent with (-)-botryodiplodin playing a role in the pathology of some, but not all, M. phaseolina isolates from soybeans with charcoal rot disease in southern USA. © 2019 by the authors

Soybean Seed Sugars: A Role in the Mechanism of Resistance to Charcoal Rot and Potential Use as Biomarkers in Selection

Charcoal rot, caused by Macrophomina phaseolina, is a major soybean disease resulting in significant yield loss and poor seed quality. Currently, no resistant soybean cultivar is available in the market and resistance mechanisms to charcoal rot are unknown, although the disease is believed to infect plants from infected soil through the roots by unknown toxin-mediated mechanisms. The objective of this research was to investigate the association between seed sugars (sucrose, raffinose, stachyose, glucose, and fructose) and their role as biomarkers in the soybean defense mechanism in the moderately resistant (MR) and susceptible (S) genotypes to charcoal rot. Seven MR and six S genotypes were grown under irrigated (IR) and non-irrigated (NIR) conditions. A two-year field experiment was conducted in 2012 and 2013 at Jackson, TN, USA. The main findings in this research were that MR genotypes generally had the ability to maintain higher seed levels of sucrose, glucose, and fructose than did S genotypes. Conversely, susceptible genotypes showed a higher level of stachyose and lower levels of sucrose, glucose, and fructose. This was observed in 6 out of 7 MR genotypes and in 4 out of 6 S genotypes in 2012; and in 5 out of 7 MR genotypes and in 5 out of 6 S genotypes in 2013. The response of S genotypes with higher levels of stachyose and lower sucrose, glucose, and fructose, compared with those of MR genotypes, may indicate the possible role of these sugars in a defense mechanism against charcoal rot. It also indicates that nutrient pathways in MR genotypes allowed for a higher influx of nutritious sugars (sucrose, glucose, and fructose) than did S genotypes, suggesting these sugars as potential biomarkers for selecting MR soybean plants after harvest. This research provides new knowledge on seed sugars and helps in understanding the impact of charcoal rot on seed sugars in moderately resistant and susceptible genotypes

Advanced power plant training simulator for VVER-440/V230 nuclear power plants

An advanced, workstation based, nuclear power plant simulator has been developed for use in training the operational staff of the Bohunice Nuclear Power Plant. This training simulator uses state-of- the-art computer hardware and software and provides the capability to simultaneously include six members of the power plant operating staff in the training sessions. A detailed reactor model has been developed, representing the Bohunice VVER-44O/V230 plants, for use with the RELAP5 simulation software. In addition, a comprehensive validation program has been completed that compares the simulation results of the advanced simulator with the results from a current VVER-44O/V230 simulator. A summary of the training features and capabilities of the simulator is also provided

Analysis of a SBLOCA initiated by an ATWS event

The response of a four-loop Westinghouse pressurized water reactor to SBLOCAs initiated as a result of an anticipated transient without scram (ATWS) has been analyzed using the RELAP5 computer code. The ATWS is initiated by a loss-of-feedwater, and the small breaks were due to either one or three stuck-open safety valves or reactor coolant pump seal failure. For the cases analyzed, the results show that a LOF-ATWS followed by a SBLOCA does not have more safety significance than that found when each accident is analyzed independently of one another

Effect of Increased Nitrogen Application Rates and Environment on Protein, Oil, Fatty Acids, and Minerals in Sesame (Sesamum indicum) Seed Grown under Mississippi Delta Conditions

Information on the effect of nitrogen fertilizer rates and environment on sesame seed composition and nutrition is scarce. The objective of this research was to investigate the effects of nitrogen fertilizer application rates on sesame seed yield, protein, oil, fatty acids, and mineral nutrition. A two-year (2014, 2015) field experiment was conducted. Nitrogen fertilizer (urea ammonium nitrate) solution (UAN, 32% N) was applied by side dressing to four sesame varieties (S-34, S-35, S-38, S-39) at rates of 44.7, 67.2, 89.6 and 112.0 kg\ub7ha-1. Rate of 44.7 kg\ub7ha-1 was used as control since this rate is traditionally recommended in the region. Increasing nitrogen application rates resulted in higher protein and oleic acid contents in two varieties in 2014, and in all varieties in 2015. Increased protein and oleic acid were accompanied by lower total oil and linoleic acid. Increased nitrogen application also resulted in higher seed N, S, B, Cu, Fe, and Zn in 2014 in S-34 and S-35, but either a decline or no clear change was observed in seed levels of these nutrients in S-38 and S-39. In 2015, increased nitrogen application resulted in significantly higher seed N in all varieties, and higher S, B, Cu, Fe, and Zn in some varieties. A significant positive correlation was observed between nitrogen application rate and yield, and with seed levels of protein, oleic, acid, N, B, Cu, Fe, and Zn. A significant negative correlation was observed between nitrogen application rate and seed oil and linoleic acid. Thus, increased nitrogen fertilizer application resulted in higher seed protein, oleic acid, and some mineral nutrients, but lower oil and linoleic acid. However, this effect depended on variety and environmental conditions. Because higher protein and oleic acid are desirable traits for sesame seed nutritional value and oil stability, regional breeders should select sesame varieties for efficient fertilizer response

Field studies on the deterioration of microplastic films from ultra-thin compostable bags in soil

In recent years, some countries have replaced single-use plastic bags with bags manufactured from compostable plastic film that can be used for collecting food wastes and composted together with the waste. Because industrial compost contains undeteriorated fragments of these bags, application to field soil is a potential source of small-sized residues from these bags. This study was undertaken to examine deterioration of these compostable film microplastics (CFMPs) in field soil at three different localities in Italy. Deterioration of CFMPs did not exceed 5.7% surface area reduction during the 12-month experimental period in two sites located in Northern Italy. More deterioration was observed in the Southern site, with 7.2% surface area reduction. Deterioration was significantly increased when fields were amended with industrial compost (up to 9.6%), but not with home compost. Up to 92.9% of the recovered CFMPs were associated with the soil fungus Aspergillus flavus, with 20.1%–71.2% aflatoxin-producing isolates. Application of industrial compost resulted in a significant increase in the percentage of CFMPs associated with A. flavus. This observation provides an argument for government regulation of accumulation of CFMPs and elevation of hazardous fungi levels in agricultural soils that receive industrial compost

Use of a biochar-based formulation for coating corn seeds

The series of experiments summarized here were conducted with the objective to evaluate the benefits of using biochar for coating corn seeds. Seeds coated with a slurry containing bio-based ingredients and biochar were tested for germination and vigor, and for their potential to being infected by the fungus Aspergillus flavus, using a novel single seed incubator specifically designed for these purposes. Biochar-treated seeds were also planted for two years in experimental fields in the Mississippi Delta to evaluate their effect on corn yield and aflatoxin contamination of kernels. Experiments were conducted with two types of commercial biochar; one was obtained from hardwood residues and the other from coconut shells. Application of both types of biochar for coating the seeds did not affect seed germination and vigor. However, treated seeds showed increased wettability and a more rapid water uptake. This resulted in a 8.5% shortening of germination time. Microbiological analysis using plate culturing and qPCR methods showed that biochar was not conducive to the growth of A. flavus. This was also confirmed by analyzing soil samples that were collected from experimental fields located in the Mississippi Delta. Most importantly, although aflatoxin contamination was different in the two experimental years, aflatoxin contamination of corn kernels was not affected by biochar-based formulations