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

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

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

Utilisation of an operative difficulty grading scale for laparoscopic cholecystectomy

Background A reliable system for grading operative difficulty of laparoscopic cholecystectomy would standardise description of findings and reporting of outcomes. The aim of this study was to validate a difficulty grading system (Nassar scale), testing its applicability and consistency in two large prospective datasets. Methods Patient and disease-related variables and 30-day outcomes were identified in two prospective cholecystectomy databases: the multi-centre prospective cohort of 8820 patients from the recent CholeS Study and the single-surgeon series containing 4089 patients. Operative data and patient outcomes were correlated with Nassar operative difficultly scale, using Kendall’s tau for dichotomous variables, or Jonckheere–Terpstra tests for continuous variables. A ROC curve analysis was performed, to quantify the predictive accuracy of the scale for each outcome, with continuous outcomes dichotomised, prior to analysis. Results A higher operative difficulty grade was consistently associated with worse outcomes for the patients in both the reference and CholeS cohorts. The median length of stay increased from 0 to 4 days, and the 30-day complication rate from 7.6 to 24.4% as the difficulty grade increased from 1 to 4/5 (both p < 0.001). In the CholeS cohort, a higher difficulty grade was found to be most strongly associated with conversion to open and 30-day mortality (AUROC = 0.903, 0.822, respectively). On multivariable analysis, the Nassar operative difficultly scale was found to be a significant independent predictor of operative duration, conversion to open surgery, 30-day complications and 30-day reintervention (all p < 0.001). Conclusion We have shown that an operative difficulty scale can standardise the description of operative findings by multiple grades of surgeons to facilitate audit, training assessment and research. It provides a tool for reporting operative findings, disease severity and technical difficulty and can be utilised in future research to reliably compare outcomes according to case mix and intra-operative difficulty

Functional Analysis of an Acid Adaptive DNA Adenine Methyltransferase from Helicobacter pylori 26695

HP0593 DNA-(N6-adenine)-methyltransferase (HP0593 MTase) is a member of a Type III restriction-modification system in Helicobacter pylori strain 26695. HP0593 MTase has been cloned, overexpressed and purified heterologously in Escherichia coli. The recognition sequence of the purified MTase was determined as 5′-GCAG-3′and the site of methylation was found to be adenine. The activity of HP0593 MTase was found to be optimal at pH 5.5. This is a unique property in context of natural adaptation of H. pylori in its acidic niche. Dot-blot assay using antibodies that react specifically with DNA containing m6A modification confirmed that HP0593 MTase is an adenine-specific MTase. HP0593 MTase occurred as both monomer and dimer in solution as determined by gel-filtration chromatography and chemical-crosslinking studies. The nonlinear dependence of methylation activity on enzyme concentration indicated that more than one molecule of enzyme was required for its activity. Analysis of initial velocity with AdoMet as a substrate showed that two molecules of AdoMet bind to HP0593 MTase, which is the first example in case of Type III MTases. Interestingly, metal ion cofactors such as Co2+, Mn2+, and also Mg2+ stimulated the HP0593 MTase activity. Preincubation and isotope partitioning analyses clearly indicated that HP0593 MTase-DNA complex is catalytically competent, and suggested that DNA binds to the MTase first followed by AdoMet. HP0593 MTase shows a distributive mechanism of methylation on DNA having more than one recognition site. Considering the occurrence of GCAG sequence in the potential promoter regions of physiologically important genes in H. pylori, our results provide impetus for exploring the role of this DNA MTase in the cellular processes of H. pylori

Mammal responses to global changes in human activity vary by trophic group and landscape

Wildlife must adapt to human presence to survive in the Anthropocene, so it is critical to understand species responses to humans in different contexts. We used camera trapping as a lens to view mammal responses to changes in human activity during the COVID-19 pandemic. Across 163 species sampled in 102 projects around the world, changes in the amount and timing of animal activity varied widely. Under higher human activity, mammals were less active in undeveloped areas but unexpectedly more active in developed areas while exhibiting greater nocturnality. Carnivores were most sensitive, showing the strongest decreases in activity and greatest increases in nocturnality. Wildlife managers must consider how habituation and uneven sensitivity across species may cause fundamental differences in human–wildlife interactions along gradients of human influence.Peer reviewe

Population‐based cohort study of outcomes following cholecystectomy for benign gallbladder diseases

Background The aim was to describe the management of benign gallbladder disease and identify characteristics associated with all‐cause 30‐day readmissions and complications in a prospective population‐based cohort. Methods Data were collected on consecutive patients undergoing cholecystectomy in acute UK and Irish hospitals between 1 March and 1 May 2014. Potential explanatory variables influencing all‐cause 30‐day readmissions and complications were analysed by means of multilevel, multivariable logistic regression modelling using a two‐level hierarchical structure with patients (level 1) nested within hospitals (level 2). Results Data were collected on 8909 patients undergoing cholecystectomy from 167 hospitals. Some 1451 cholecystectomies (16·3 per cent) were performed as an emergency, 4165 (46·8 per cent) as elective operations, and 3293 patients (37·0 per cent) had had at least one previous emergency admission, but had surgery on a delayed basis. The readmission and complication rates at 30 days were 7·1 per cent (633 of 8909) and 10·8 per cent (962 of 8909) respectively. Both readmissions and complications were independently associated with increasing ASA fitness grade, duration of surgery, and increasing numbers of emergency admissions with gallbladder disease before cholecystectomy. No identifiable hospital characteristics were linked to readmissions and complications. Conclusion Readmissions and complications following cholecystectomy are common and associated with patient and disease characteristics