Rapid identification of peanut seed quality is crucial for public health. In this study, we present a terahertz wave imaging system using a convolutional neural network (CNN) machine learning approach. Terahertz waves are capable of penetrating the seed shell to identify the quality of peanuts without causing any damage to the seeds. The specificity of seed quality on terahertz wave images is investigated, and the image characteristics of five different qualities are summarized. Terahertz wave images are digitized and used for training and testing of convolutional neural networks, resulting in a high model accuracy of 98.7% in quality identification. The trained THz-CNNs system can accurately identify standard, mildewed, defective, dried and germinated seeds, with an average detection time of 2.2 s. This process does not require any sample preparation steps such as concentration or culture. Our method swiftly and accurately assesses shelled seed quality non-destructively.

Fungal infections are becoming a severe threat to the security of global public health due to the extensive use of antibiotic medications and the rise in immune-deficient patients globally. Additionally, there is an increase in the development of fungus resistance to available antifungal medications. It is necessary to focus on the development of new antifungal medications in order to address these problems. The wide range of chemical structures, low cost, high availability, high antimicrobial action, and lack of adverse effects are the characteristics of plant secondary metabolites. In order to find and develop new antifungal medications, plant secondary metabolites like glucosinolate (GSL) derivatives are crucial sources of information. These natural compounds are enzymatically transformed into isothiocyanates (ITCs), nitriles, epithionitriles, oxazolidin-2-thion, and thiocyanate when they get mechanically damaged. The current review offers a thorough understanding of how isothiocyanates affect fungi with detailed mechanism. Along with this antifungal activity of nitriles, epithionitriles, oxazolidin-2-thion, and thiocyanate are mentioned. The review summarizes our present understanding of the following subjects: role of isothiocyanate by inhibiting aflatoxin biosynthesis, effect of isothiocyanate on transcriptomes, isothiocyanate targets cell membrane, role of isothiocyanate in efflux, and the role of isothiocyanate in synergistic activity. Antifungal activity of nitrile, epithionitrile, oxazolidine-2-thion, and thiocyanate is mentioned. Cytotoxicity study and clinical trials data were also added. More extensive studies will be needed in this field to assess safety concerns and clinical efficacies of GSL derivatives.

Aflatoxins are carcinogens that can contaminate food and affect various body organs especially liver and kidney. When consumed, aflatoxin B1 (AFB1) is partially metabolised into aflatoxin M1 (AFM1), which is excreted in the urine.Breast milk may also contain AFM1 due to maternal dietary intake from contaminated food. This cross-sectional study aimed to determine the levels of AFM1 in both urine and breast milk among breastfeeding mothers (n = 256). The mother\'s demographic information was collected during recruitment. Mothers were then scheduled for an appointment to provide a morning urine sample along with five to ten mL samples of breast milk. AFM1 levels in both samples were analysed using an enzyme-linked immunosorbent assay (ELISA). Spearman\'s rho and Chi-square were used to determine the associations between mean levels of AFM1 in urine and breast milk. Findings show 68.0% of urine samples were contaminated with AFM1 (mean levels = 0.08 ± 0.04 ng/mL), while 14.8% of breast milk samples had AFM1 (mean levels = 5.94 ± 1.81 ng/kg). Urine AFM1 levels were not significantly associated with AFM1 levels in breast milk (p > 0.05). This study can act as a baseline for future research examining long-term aflatoxin exposure among both mothers and infants.

The potential of Raman hyperspectral imaging with a 785 nm excitation line laser was examined for the detection of aflatoxin contamination in corn kernels. Nine-hundred kernels were artificially inoculated in the laboratory, with 300 kernels each inoculated with AF13 (aflatoxigenic) fungus, AF36 (nonaflatoxigenic) fungus, and sterile distilled water (control). One-hundred kernels from each treatment were subsequently incubated for 3, 5, and 8 days. The mean spectra of single kernels were extracted from the endosperm side and the embryo area of the germ side, and local Raman peaks were identified based upon the calculated reference spectra of aflatoxin-negative and -positive categories separately. The principal component analysis-linear discriminant analysis models were established using different types of variable inputs including original full spectra, preprocessed full spectra, and identified local peaks over kernel endosperm-side, germ-side, and both sides. The results of the established discriminant models showed that the germ-side spectra performed better than the endosperm-side spectra. Based upon the 20 ppb-threshold, the best mean prediction accuracy of 82.6% was achieved for the aflatoxin-negative category using the original spectra in the combined form of both kernel sides, and the best mean prediction accuracy of 86.7% was obtained for the -positive category using the preprocessed germ-side spectra. Based upon the 100 ppb-threshold, the best mean prediction accuracies of 85.0% and 89.6% were achieved for the aflatoxin-negative and -positive categories separately, using the same type of variable inputs for the 20 ppb-threshold. In terms of overall prediction accuracy, the models established upon the original spectra in the combined form of both kernel sides achieved the best predictive performance, regardless of the threshold. The mean overall prediction accuracies of 81.8% and 84.5% were achieved with the 20 ppb- and 100 ppb-thresholds, respectively.

Aflatoxins (AFs) are highly toxic mycotoxins produced by the fungi, Aspergillus flavus and Aspergillus parasiticus. AFs pose severe health risks owing to their acute toxicity and carcinogenic properties. The control of AF contamination remains significantly challenging despite the extensive efforts toward controlling it. Here, we investigated the potential of mushroom extracts as a source of AF biosynthetic inhibitors. The A. parasiticus mutant strain, NFRI-95, that accumulates an AF biosynthesis intermediate, norsolorinic acid, was used in the bioassay to detect the inhibitory activity against AF biosynthesis. The screening of 195 mushroom extracts revealed that the culture filtrate extract of Chondrostereum purpureum exhibited strong inhibitory activity against AF biosynthesis. Next, large-scale culturing of C. purpureum was performed to isolate the compounds accounting for the inhibitory activity. The culture filtrate was extracted with ethyl acetate, after which the active compound was isolated by silica gel column chromatography and preparative high performance liquid chromatography (HPLC). The active compound was identified as cyclo(Val-Pro) by spectroscopic analyses. Further, four stereoisomers of cyclo(Val-Pro) were synthesized by the condensation of the N-Boc derivatives of d- and l-valine with the methyl esters of d- and l-proline. The naturally isolated compound was identified as cyclo(l-Val-l-Pro) by comparing its retention time with those of synthetic compounds by chiral HPLC analysis and CD spectra. The IC50 value of cyclo(L-Val-L-Pro) was 2.4 mM, whereas the LD, DL, and DD isomers exhibited weaker activities, with IC50 values of >5 mM.

Non-genetic variation limits the identification of novel maize germplasm with genetic markers for reduced Aspergillus flavus infection and aflatoxin contamination. Aflatoxin measurements can vary substantially within fields containing the same germplasm following inoculation with A. flavus. While some variation is expected due to microenvironmental differences, components of field screening methodologies may also contribute to variability in collected data. Therefore, the objective of this study is to test the effects of three different shelling methods (whole ear (WE), ear end removal (EER), and inoculation site-surrounding (ISS)) to obtain bulk samples from maize on aflatoxin measurements. Five ears per row of three inbred lines and two hybrids were inoculated with A. flavus, then shelled using the three different methods, and aflatoxin was quantified. Overall, EER and ISS resulted in reduced coefficients of variance (CVs) in comparison to WE for both inbred and hybrid maize lines, with two exceptions. Susceptible B73 showed increased CVs with both EER and ISS compared to WE, and resistant Mp719\'s EER CVs marginally increased compared to WE. While WE is the standard practice for most breeding programs due to its technical simplicity, EER and ISS may allow for finely phenotyping parental lines for further breeding applications.

Chemical pesticides help reduce crop loss during production and storage. However, the carbon footprints and ecological costs associated with this strategy are unsustainable. Here, we used three in vitro models to characterize how different Trichoderma species interact with two aflatoxin producers, Aspergillus flavus and Aspergillus parasiticus, to help develop a climate-resilient biological control strategy against aflatoxigenic Aspergillus species. The growth rate of Trichoderma species is a critical factor in suppressing aflatoxigenic strains via physical interactions. The dual plate assay suggests that Trichoderma mainly suppresses A. flavus via antibiosis, whereas the suppression of A. parasiticus occurs through mycoparasitism. Volatile organic compounds (VOCs) produced by Trichoderma inhibited the growth of A. parasiticus (34.6 ± 3.3%) and A. flavus (20.9 ± 1.6%). The VOCs released by T. asperellum BTU and T. harzianum OSK-34 were most effective in suppressing A. flavus growth. Metabolites secreted by T. asperellum OSK-38, T. asperellum BTU, T. virens OSK-13, and T. virens OSK-36 reduced the growth of both aflatoxigenic species. Overall, T. asperellum BTU was the most effective at suppressing the growth and aflatoxin B1 production of both species across all models. This work will guide efforts to screen for effective biological control agents to mitigate aflatoxin accumulation.

Aspergillus flavus produces aflatoxin, a carcinogenic fungal toxin that poses a threat to the agricultural and food industries. There is a concern that the distribution of aflatoxin-producing A. flavus is expanding in Japan due to climate change, and it is necessary to understand what types of strains inhabit. In this study, we sequenced the genomes of four Aspergillus strains isolated from agricultural fields in the Ibaraki prefecture of Japan and identified their genetic variants. Phylogenetic analysis based on single-nucleotide variants revealed that the two aflatoxin-producing strains were closely related to A. flavus NRRL3357, whereas the two non-producing strains were closely related to the RIB40 strain of Aspergillus oryzae, a fungus widely used in the Japanese fermentation industry. A detailed analysis of the variants in the aflatoxin biosynthetic gene cluster showed that the two aflatoxin-producing strains belonged to different morphotype lineages. RT-qPCR results indicated that the expression of aflatoxin biosynthetic genes was consistent with aflatoxin production in the two aflatoxin-producing strains, whereas the two non-producing strains expressed most of the aflatoxin biosynthetic genes, unlike common knowledge in A. oryzae, suggesting that the lack of aflatoxin production was attributed to genes outside of the aflatoxin biosynthetic gene cluster in these strains.

Consumption of milk is linked to improved nutrient intake and reduced risk of child malnutrition in low and middle-income countries. However, these benefits are contingent on the safety and quality of the milk. Milk consumption may alleviate the widespread risk of malnutrition in rural Ethiopia, but milk-borne contaminants may also compromise child health. We aimed to: i) identify the types of dairy feeds used, their storage conditions, and potential risk of aflatoxin contamination; ii) assess stakeholders\' knowledge about aflatoxin contamination along the value chain; and iii) assess parental practices on feeding milk to infants and young children. This qualitative study was conducted in the Sidama region, southern Ethiopia. In-depth interviews (n = 12) and key-informant interviews (n = 18) were conducted with actors along the dairy value chain. Focus-group discussions were conducted with farmers (9FGD/n = 129) and child caregivers (9FGD/n = 122). Study participants were selected to represent a rural-urban gradient, as well as low- and high- dairy cow holdings. We found that while animal-feed processors and their distribution agents had relatively good knowledge about aflatoxin, farmers and retailers did not. Feed storage conditions were poor. Many respondents linked moldy feeds to animal health but not to human health. Farmers\' feed choice was influenced by cost, seasonality, and herd size. Small-holding farmers had limited access to commercial feed. Children\'s consumption of milk was limited to skim milk, as butter was extracted and sold for income. The high cost of dairy products also led some parents to dilute skim milk with water before feeding children, compromising the nutritional value and safety of the milk. Our findings underscore the need to address the gaps in aflatoxin and food safety knowledge, improve storage conditions, and ensure the availability of quality feed to increase the sector\'s productivity, but most importantly to protect consumers\' health and well-being, especially infants and young children.

The target of rapamycin (TOR) signaling pathway is highly conserved and plays a crucial role in diverse biological processes in eukaryotes. Despite its significance, the underlying mechanism of the TOR pathway in Aspergillus flavus remains elusive. In this study, we comprehensively analyzed the TOR signaling pathway in A. flavus by identifying and characterizing nine genes that encode distinct components of this pathway. The FK506-binding protein Fkbp3 and its lysine succinylation are important for aflatoxin production and rapamycin resistance. The TorA kinase plays a pivotal role in the regulation of growth, spore production, aflatoxin biosynthesis, and responses to rapamycin and cell membrane stress. As a significant downstream effector molecule of the TorA kinase, the Sch9 kinase regulates aflatoxin B1 (AFB1) synthesis, osmotic and calcium stress response in A. flavus, and this regulation is mediated through its S_TKc, S_TK_X domains, and the ATP-binding site at K340. We also showed that the Sch9 kinase may have a regulatory impact on the high osmolarity glycerol (HOG) signaling pathway. TapA and TipA, the other downstream components of the TorA kinase, play a significant role in regulating cell wall stress response in A. flavus. Moreover, the members of the TapA-phosphatase complexes, SitA and Ppg1, are important for various biological processes in A. flavus, including vegetative growth, sclerotia formation, AFB1 biosynthesis, and pathogenicity. We also demonstrated that SitA and Ppg1 are involved in regulating lipid droplets (LDs) biogenesis and cell wall integrity (CWI) signaling pathways. In addition, another phosphatase complex, Nem1/Spo7, plays critical roles in hyphal development, conidiation, aflatoxin production, and LDs biogenesis. Collectively, our study has provided important insight into the regulatory network of the TOR signaling pathway and has elucidated the underlying molecular mechanisms of aflatoxin biosynthesis in A. flavus.