Aflatoxins in food systems: recent advances in toxicology, biosynthesis, regulation and mitigation through green nanoformulations.

Aflatoxins are hepatocarcinogenic and immunosuppressive mycotoxins mainly synthesized by Aspergillus flavus, A. parasiticus and A. nomius in food systems, causing negative health impacts to humans and other organisms. Aflatoxins contaminate most of the agri-products of tropical and subtropical regions due to hot and humid conditions and persist in food items even after food processing steps, causing major threat towards the food security. Different physical and chemical strategies have been applied to mitigate aflatoxin contamination. However, negative impacts of chemical preservatives towards health and environment limit their practical applicability. In this regard, plant-based preservatives, due to their economical, eco-friendly and safer profile, are considered as a sustainable approach towards food safety. Incorporation of nanotechnology would enhance the bio-efficacy of green preservatives by overcoming some of their major challenges, such as volatility. The present review deals with recent information on toxicology and molecular and enzymatic regulatory pathways in the biosynthesis of aflatoxins in food systems. A proper understanding of the role of different genes and regulatory proteins may provide novel preventive strategies for aflatoxin detoxification and also in development of aflatoxin-resistant food items. The review also emphasizes the role of green nanoformulations as a sustainable approach towards the management of aflatoxins in food systems. In addition, some technological challenges of green nanotechnology have also been discussed in this review, along with highlighting some future perspectives. © 2022 Society of Chemical Industry.

Emission modeling at toll plaza under mixed traffic condition using simulation.

Mixed traffic conditions, i.e., the presence of different vehicle classes in a toll lane, causes congestion, leading to extra delay and emissions. The determination of the emission levels in the field is tedious process and thus there is need to look over it. Further, the dynamic toll pricing schemes are well established in developed countries to mitigate the congestion, but its application in developing countries is not observed so far. Hence, an attempt is made in the present study to establish the dynamic toll rates based on emission at toll plazas operating under mixed traffic conditions. The sensitivity analysis is carried out by changing the traffic composition and the approach volume in simulation. After that, the traffic composition and approach volume-based emission estimation equations are developed that can be used to predict the emissions easily in the field. The methodology finally moves towards the development of emission-based toll rates (EBTR).

De Novo Genome Assembly Of Solanum Sitiens Reveals Structural Variation Associated With Drought And Salinity Tolerance.

MOTIVATION: Solanum sitiens is a self-incompatible wild relative of tomato, characterised by salt and drought resistance traits, with the potential to contribute through breeding programmes to crop improvement in cultivated tomato. This species has a distinct morphology, classification and ecotype compared to other stress resistant wild tomato relatives such as S. pennellii and S. chilense. Therefore, the availability of a reference genome for S. sitiens will facilitate the genetic and molecular understanding of salt and drought resistance. RESULTS: A high-quality de novo genome and transcriptome assembly for S. sitiens (Accession LA1974) has been developed. A hybrid assembly strategy was followed using Illumina short reads (∼159X coverage) and PacBio long reads (∼44X coverage), generating a total of ∼262 Gbp of DNA sequence. A reference genome of 1,245 Mbp, arranged in 1,483 scaffolds with a N50 of 1.826 Mbp was generated. Genome completeness was estimated at 95% using the Benchmarking Universal Single-Copy Orthologs (BUSCO) and the K-mer Analysis Tool (KAT). In addition, ∼63 Gbp of RNA-Seq were generated to support the prediction of 31,164 genes from the assembly, and to perform a de novo transcriptome. Lastly, we identified three large inversions compared to S. lycopersicum, containing several drought resistance related genes, such as beta-amylase 1 and YUCCA7. AVAILABILITY: S. sitiens (LA1974) raw sequencing, transcriptome and genome assembly have been deposited at the NCBI's Sequence Read Archive, under the BioProject number "PRJNA633104".All the commands and scripts necessary to generate the assembly are available at the following github repository: https://github.com/MCorentin/Solanum_sitiens_assembly. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Preparation and characterization of a novel nanoemulsion consisting of chitosan and Cinnamomum tamala essential oil and its effect on shelf-life lengthening of stored millets.

This study aimed to improve the stability of Cinnamomum tamala essential oil (CTEO) via encapsulating into chitosan nanoemulsion (CsNe) through an ionic-gelation technique and explore its food preservative efficacy against aflatoxigenic strain of Aspergillus flavus (AFLHPSi-1, isolated from stored millet), aflatoxin B1 (AFB1) contamination, and lipid peroxidation, causing qualitative deterioration of stored millets. The CTEO was characterized through gas chromatography-mass spectrometry (GC-MS) analysis that confirmed the presence of linalool as a major component occupying approximately 82.64% of the total oil. The synthesized nanoparticles were characterized through scanning electron microscopy (SEM), fourier transform infrared (FTIR) spectroscopy, and X-ray diffraction (XRD) analysis. The encapsulation efficiency (EE) and loading capacity (LC) of CTEO-CsNe were found to be 97.71% and 3.33%, respectively. In vitro release study showed a biphasic release pattern: with an initial burst release followed by a controlled release of CTEO. During investigation of efficacy, the CTEO-CsNe caused complete inhibition of A. flavus growth, and AFB1 biosynthesis at 1.0 and 0.8 µL/mL, respectively. The CTEO-CsNe exhibited its antifungal mode of action by altering fungal plasma membrane integrity (ergosterol inhibition) and permeability (leakage of important cellular constituents), and antiaflatoxigenic mode of action by inhibiting cellular methylglyoxal biosynthesis. CTEO-CsNe showed high free radical scavenging capacity (IC50 = 5.08 and 2.56 µL/mL) against DPPH•+ and ABTS•+ radicals, respectively. In addition, CTEO-CsNe presented remarkable preservative efficacy, inhibiting AFB1 and lipid peroxidation in model food system (Setaria italica) without altering their organoleptic properties. Based on overall results, CTEO-CsNe can be recommended as a novel shelf-life enhancer of stored millet samples.

A novel monopartite begomovirus and satellites associated with yellow mosaic disease of Sida spp. in India.

Begomoviruses (family Geminiviridae) cause severe diseases in many economically important crops and non-cultivated plants in the warmer regions of the world. Non-cultivated weeds have been reported to act as natural virus reservoirs. In January 2016, Sida plants with yellow mosaic symptoms were found at the edge of an agricultural field in Gujarat, India. Sequence analysis of the viral genomic components cloned from a diseased Sida plant indicated the presence of a distinct monopartite begomovirus (proposed as sida yellow mosaic Gujarat virus) along with a betasatellite (ludwigia leaf distortion betasatellite) and an alphasatellite (malvastrum yellow mosaic alphasatellite). Our results emphasize that this weed may harbor a begomovirus-alphasatellite-betasatellite complex. This host serves as a potential source of virus inoculum, which can be transmitted by whiteflies to other cultivated crops.

Bhendi Yellow Vein Mosaic Virus and Bhendi Yellow Vein Mosaic Betasatellite Cause Enation Leaf Curl Disease and Alter Host Phytochemical Contents in Okra.

Whitefly (Bemisia tabaci)-transmitted begomoviruses cause severe diseases in numerous economically important dicotyledonous plants. Okra enation leaf curl disease (OELCuD) has emerged as a serious threat to okra (Abelmoschus esculentus L. Moench) cultivation in the Indian subcontinent. This study reports the association of a monopartite begomovirus (bhendi yellow vein mosaic virus; BYVMV) and betasatellite (bhendi yellow vein mosaic betasatellite; BYVB) with OELCuD in the Mau region of Uttar Pradesh, India. The BYVMV alone inoculated Nicotiana benthamiana and A. esculentus cv. Pusa Sawani plants developed mild symptoms. Co-inoculation of BYVMV and BYVB resulted in a reduced incubation period, an increased symptom severity, and an enhanced BYVMV accumulation by Southern hybridization and quantitative real-time PCR. This is the first study that satisfies Koch's postulates for OELCuD in its natural host. Activities of various antioxidative enzymes were significantly increased in the virus-inoculated okra plants. Differential responses in various biochemical components (such as photosynthetic pigments, phenol, proline, and sugar) in diseased okra plants were observed. This change in phytochemical responses is significant in understanding its impact on virus pathogenesis and disease development.

Essential oils and their nanoformulations as green preservatives to boost food safety against mycotoxin contamination of food commodities: a review.

Postharvest food spoilage due to fungal and mycotoxin contamination is a major challenge in tropical countries, leading to severe adverse effects on human health. Because of the negative effects of synthetic preservatives on both human health and the environment, it has been recommended that chemicals that have a botanical origin, with an eco-friendly nature and a favorable safety profile, should be used as green preservatives. Recently, the food industry and consumers have been shifting drastically towards green consumerism because of their increased concerns about health and the environment. Among different plant-based products, essential oils (EOs) and their bioactive components are strongly preferred as antimicrobial food preservatives. Despite having potent antimicrobial efficacy and preservation potential against fungal and mycotoxin contamination, essential oils and their bioactive components have limited practical applicability caused by their high volatility and their instability, implying the development of techniques to overcome the challenges associated with EO application. Essential oils and their bioactive components are promising alternatives to synthetic preservatives. To overcome challenges associated with EOs, nanotechnology has emerged as a novel technology in the food industries. Nanoencapsulation may boost the preservative potential of different essential oils by improving their solubility, stability, and targeted sustainable release. Nanoencapsulation of EOs is therefore currently being practiced to improve the stability and bioactivity of natural products. The present review has dealt extensively with the application of EOs and their nanoformulated products encapsulated in suitable polymeric matrices, so as to recommend them as novel green preservatives against foodborne molds and mycotoxin-induced deterioration of stored food commodities. © 2021 Society of Chemical Industry.

Profiling of polyphenols in phalsa (Grewia asiatica L) fruits based on liquid chromatography high resolution mass spectrometry.

The aim of the study was to screen the metabolite profile of phalsa (Grewia asiatica), an underutilized fruit crop, using liquid chromatography-high resolution mass spectrometric analysis. A total of 50 compounds were tentatively identified based on their molecular mass and characteristic fragment ions, each with less than 5 ppm of mass error. These compounds included 21 flavonols, 2 dihydroflavonols, 7 flavones, 3 flavanols, 6 anthocyanins, 3 isoflavonoids, 2 phenolic acids, 2 flavanones, and 4 other phenolics. Flavonols were the predominant group of compounds, representing around 52.6% of the total phenolics. The paper has also discussed the potentiality of phalsa as an emerging functional food for the management of various human diseases in relation to the existing literature.

LC-MS/MS method optimization and validation for trace-level analysis of 71 crop protection chemicals in pulses.

An analytical method involving modified QuEChERS (quick, easy, cheap, effective, rugged and safe) technique coupled with LC-MS/MS (liquid chromatography-tandem mass spectroscopy) has been developed for simultaneous identification and quantification of 71 pesticides (insecticides/acaricides/nematicides (32), fungicides (12), herbicides (26), plant growth regulator (1)) in different pulses (edible seeds of leguminous plants), namely gram, lentil, black gram and pea. The analysis was done using Shimadzu LC-MS/MS-8030 instrument equipped with Zorbax Eclipse Plus C-18 column operating under electrospray ionization (ESI) in positive and negative modes. Validation of method was done as per a single laboratory validation approach. Nine-point linear calibration curves for each pesticide were obtained in the range of 0.005 to 2 µg/g with correlation coefficient of ≥ 0.98. Limit of detection (LOD) for all the pesticides was achieved in the range of 0.001-0.015 µg/g, whereas the limit of quantification (LOQ) were found in the range of 0.01-0.05 µg/g. Recovery studies were conducted at 0.01- and 0.05-µg/g level of fortification using modified buffered QuEChERS method standardized for low moisture foods. It was observed that due to matrix interference, only around 75% of the pesticides were recovered in an acceptable range of 70-120% when compared against the solvent standard. With matrix-matched standards, 95% of the recovery results came within acceptable range with highly acceptable HorRat ratio (between 0.2 and 0.8) indicating satisfactory precision. The global and expanded uncertainties for estimation of each pesticide in above-mentioned pulse matrices were calculated to evaluate the suitability of the developed method for quantification of pesticides in pulse matrices.

Genetic diversity of cucumber estimated by morpho-physiological and EST-SSR markers.

In the present study, genetic variation among 40 cucumber genotypes was analyzed by means of morpho-physiological traits and 21 EST-SSR markers. Diversity was observed for morpho-physiological characters like days to 50% female flowering (37-46.9, number of fruits/plant (1.33-5.80), average fruit weight (41-333), vine length (36-364), relative water content (58.5-92.7), electrolyte leakage (15.9-37.1), photosynthetic efficiency (0.40-0.75) and chlorophyll concentration index (11.1-28.6). The pair wise Jaccard similarity coefficient ranged from 0.00 to 0.27 for quantitative traits and 0.24 to 0.96 for EST-SSR markers indicating that the accessions represent genetically diverse populations. With twenty-one EST-SSR markers, polymorphism revealed among 40 cucumber genotypes, number of alleles varied 2-6 with an average 3.05. Polymorphism information content varied from 0.002 to 0.989 (mean = 0.308). The number of effective allele (Ne), expected heterozygosity (He) and unbiased expected heterozygosity (uHe) of these EST-SSRs were 1.079-1.753, 0.074-0.428 and 0.074-0.434, respectively. Same 21 EST-SSR markers transferability checked in four other Cucumis species: snapmelon (Cucumis melo var. momordica), muskmelon (Cucumis melo L.), pickling melon (Cucumis melo var. conomon) and wild muskmelon (Cucumis melo var. agrestis) with frequency of 61.9, 95.2, 76.2, and 76.2%, respectively. Present study provides useful information on variability, which can assist geneticists with desirable traits for cucumber germplasm utilization. Observed physiological parameters may assists in selection of genotype for abiotic stress tolerance also, EST-SSR markers may be useful for genetic studies in related species.

Chitosan nanoemulsion incorporated with Carum carvi essential oil as ecofriendly alternative for mitigation of aflatoxin B1 contamination in stored herbal raw materials.

The present investigation entails the first report on entrapment of Carum carvi essential oil (CCEO) into chitosan polymer matrix for protection of stored herbal raw materials against fungal inhabitation and aflatoxin B1 (AFB1) production. Physico-chemical characterization of nanoencapsulated CCEO was performed through Fourier transform infrared spectroscopy, dynamic light scattering, X-ray diffractometry, and scanning electron microscopy. The nanoencapsulated CCEO displayed improved antifungal and AFB1 suppressing potentiality along with controlled delivery over unencapsulated CCEO. The encapsulated CCEO nanoemulsion obstructed the ergosterol production and escalated the efflux of cellular ions, thereby suggesting plasma membrane as prime target of antifungal action in Aspergillus flavus cells. The impairment in methyglyoxal production and modeling based carvone interaction with Afl-R protein validated the antiaflatoxigenic mechanism of action. In addition, CCEO displayed augmentation in antioxidant potentiality after encapsulation into chitosan nanomatrix. Moreover, the in-situ study demonstrated the effective protection of Withania somnifera root samples (model herbal raw material) against fungal infestation and AFB1 contamination along with prevention of lipid peroxidation. The acceptable organoleptic qualities of W. somnifera root samples and favorable safety profile in mice (animal model) strengthen the application of nanoencapsulated CCEO emulsion as nano-fungitoxicant for preservation of herbal raw materials against fungi and AFB1 mediated biodeterioration.

Green facile synthesis of cajuput (Melaleuca cajuputi Powell.) essential oil loaded chitosan film and evaluation of its effectiveness on shelf-life extension of white button mushroom.

This study reports first time investigation on efficacy of cajuput essential oil loaded chitosan nanoparticle (CjEO-CSNP) on shelf-life of white button mushroom (Agaricus bisporus) stored at 4±1 °C for 7-days. CjEO-CSNP was characterized through scanning electron microscopy, X-ray diffraction, and dynamic light scattering. The nanoparticles exhibited spherical shapes with average particle size 43.17-97.03 nm. The nanoencapsulation efficiency and loading capacity were ranged between 45.86 and 92.26% and 0.69-8.87%, respectively. The release study confirmed that CjEO-CSNP showed biphasic release patterns at different pH. Positive results were unveiled when the effect of CjEO-CSNP on shelf-life of mushroom was validated by analyzing the visual appearance and firmness. Further, CjEO-CSNP prevented weight loss and respiration rate, and improved the antioxidant activity of mushrooms. CjEO-CSNP also exhibited high safety profile (LD50= > 1200 mg/Kg body weight) without altering the sensory quality of coated mushrooms. Overall, CjEO-CSNP might be used as promising candidate to lengthen the shelf-life of button mushroom.

Development, validation and a GAPI greenness assessment for the determination of 103 pesticides in mango fruit drink using LC-MS/MS.

A robust method was developed using LC-ESI-MS/MS-based identification and quantification of 103 fortified pesticides in a mango fruit drink. Variations in QuEChERS extraction (without buffer, citrate, and/or acetate buffered) coupled with dispersive clean-up combinations were evaluated. Results showed 5 mL dilution and citrate buffered QuEChERS extraction with anhydrous (anhy) MgSO4 clean-up gave acceptable recovery for 100 pesticides @ 1 µg mL-1 fortification. The method was validated as per SANTE guidelines (SANTE/11813/2021). 95, 91, and 77 pesticides were satisfactorily recovered at 0.1, 0.05, and 0.01 µg mL-1 fortification with HorRat values ranging from 0.2-0.8 for the majority. The method showed matrix enhancement for 77 pesticides with a global uncertainty of 4.72%-23.89%. The reliability of the method was confirmed by real sample analysis of different brands of mango drinks available in the market. The greenness assessment by GAPI (Green Analytical Procedure Index) indicated the method was much greener than other contemporary methods.

Diverse begomovirus-betasatellite complexes cause tomato leaf curl disease in the western India.

In the Indian sub-continent, tomato leaf curl disease (ToLCD) of tomato caused by begomoviruses has emerged as a major limiting factor for tomato cultivation. Despite the spread of this disease in the western India, a systematic study on the characterization of virus complexes with ToLCD is lacking. Here, we report the identification of a complex of begomoviruses including 19 DNA-A and 4 DNA-B as well as 15 betasatellites with ToLCD in the western part of the country. Additionally, a novel betasatellite and an alphasatellite were also identified. The recombination breakpoints were detected in the cloned begomoviruses and betasatellites. The cloned infectious DNA constructs cause disease on the tomato (a moderately virus-resistant cultivar) plants, thus fulfilling Koch's postulates for these virus complexes. Further, the role of non-cognate DNA B/betasatellite with ToLCD-associated begomoviruses on disease development was demonstrated. It also emphasizes the evolutionary potential of these virus complexes in breaking disease resistance and plausible expansion of its host range. This necessitates to investigate the mechanism of the interaction between resistance breaking virus complexes and the infected host.

Recent trends in the nanozeolites-based oxygen concentrators and their application in respiratory disorders.

Medical-grade oxygen is the basic need for all medical complications, especially in respiratory-based discomforts. There was a drastic increase in the demand for medical-grade oxygen during the current pandemic. The non-availability of medical-grade oxygen led to several complications, including death. The oxygen concentrator was only the last hope for the patient during COVID-19 pandemic around the globe. The demands also are everlasting during other microbial respiratory infections. The yield of oxygen using conventional molecular zeolites in the traditional oxygen concentrator process is less than the yield noticed when its nano-form is used. Nanotechnology has enlightened hope for the efficient production of oxygen by such oxygen concentrators. Here in the current review work, the authors have highlighted the basic structural features of oxygen concentrators along with the current working principle. Besides, it has been tried to bridge the gap between conventional oxygen concentrators and advanced ones by using nanotechnology. Nanoparticles being usually within 100 nm in size have a high surface area to volume ratio, which makes them suitable adsorbents for oxygen. Here authors have suggested the use of nano zeolite in place of molecular zeolites in the oxygen concentrator for efficient delivery of oxygen by the oxygen concentrators.

High speed homogenization assisted encapsulation of synergistic essential oils formulation: Characterization, in vitro release study, safety profile, and efficacy towards mitigation of aflatoxin B1 induced deterioration in rice samples.

Application of essential oils to mitigate aflatoxin B1 (AFB1) contamination in food is a current research hotspot; however, their direct incorporation may cause toxic effects, and changes in food organoleptic properties. This work aimed to synthesize novel synergistic formulation of Pinus roxburghii, Juniperus communis, and Cupressus sempervirens essential oils by mixture design assay (PJC) and encapsulation of PJC formulation into chitosan nanocomposite (Nm-PJC) with an aim to protect stored rice (Oryza sativa L., prime staple food) against fungi and AFB1 mediated loss of valuable minerals, macronutrients, and fatty acids. Nm-PJC was characterized through DLS, SEM, FTIR, and XRD analyses, along with controlled delivery from chitosan nanobiopolymer. Encapsulation of synergistic formulation into chitosan-nanomatrix improved antifungal (4.0 µL/mL), antiaflatoxigenic (3.5 µL/mL), and antioxidant activities (P < 0.05). Impairment in ergosterol and methylglyoxal biosynthesis along with in-silico-homology-modeling of major components with Ver-1 and Omt-A proteins advocated chemico-molecular interaction responsible for fungal growth inhibition and AFB1 secretion. In addition, in-situ efficacy against lipid-peroxidation, fatty acid biodeterioration, and preservation of minerals, macronutrients without affecting organoleptic attributes in rice and high mammalian safety profile (9874.23 µL/kg) suggested practical application of synergistic nanoformulation as innovative smart, and green candidate to mitigate AFB1 contamination, and shelf-life extension of stored food products.

Chemically characterized nanoencapsulated Homalomena aromatica Schott. essential oil as green preservative against fungal and aflatoxin B1 contamination of stored spices based on in vitro and in situ efficacy and favorable safety profile on mice.

Present study deals with the efficacy of nanoencapsulated Homalomena aromatica essential oil (HAEO) as a potent green preservative against toxigenic Aspergillus flavus strain (AF-LHP-NS 7), storage fungi, AFB1, and free radical-mediated deterioration of stored spices. GC-MS analysis revealed linalool (68.51%) as the major component of HAEO. HAEO was encapsulated into chitosan nanomatrix (CS-HAEO-Ne) and characterized through SEM, FTIR, and XRD. CS-HAEO-Ne completely inhibited A. flavus growth and AFB1 biosynthesis at 1.25 µL/mL and 1.0 µL/mL, respectively in comparison to unencapsulated HAEO (1.75 µL/mL and 1.25 µL/mL, respectively). CS-HAEO-Ne caused significant reduction in ergosterol content in treated A. flavus and provoked leakage of cellular ions (Ca+2, Mg+2, and K+) as well as 260 nm and 280 nm absorbing materials. Depletion of methylglyoxal level in treated A. flavus cells illustrated the novel antiaflatoxigenic efficacy of CS-HAEO-Ne. CS-HAEO-Ne exhibited superior antioxidant efficacy (IC50 (DPPH) = 4.5 µL/mL) over unencapsulated HAEO (IC50 (DPPH) = 15.9 µL/mL) and phenolic content. CS-HAEO-Ne depicted excellent in situ efficacy by inhibiting fungal infestation, AFB1 contamination, lipid peroxidation, and mineral loss with acceptable sensorial profile. Moreover, broad safety paradigm (LD50 value = 7150.11 mg/kg) of CS-HAEO-Ne also suggests its application as novel green preservative to enhance shelf life of stored spices.

Chitosan encompassed Aniba rosaeodora essential oil as innovative green candidate for antifungal and antiaflatoxigenic activity in millets with emphasis on cellular and its mode of action.

The present study demonstrates first time investigation on encapsulation of Aniba rosaeodora essential oil into chitosan nanoemulsion (AREO-CsNe) with the aim of improvement of its antifungal, and aflatoxin B1 (AFB1) inhibitory performance in real food system. The GC-MS analysis of AREO revealed the presence of linalool (81.46%) as a major component. The successful encapsulation of EO into CsNe was confirmed through SEM, FTIR, and XRD analysis. The in-vitro release study showed the controlled release of AREO. AREO-CsNe caused complete inhibition of Aspergillus flavus (AFLHPSi-1) growth and AFB1 production at 0.8 and 0.6 µl/ml, respectively, which was far better than AREO (1.4 and 1.2 µl/ml, respectively). Impairment of ergosterol biosynthesis coupled with enhancement of cellular materials leakage confirmed plasma membrane as the possible antifungal target of both AREO and AREO-CsNe. Significant inhibition of methylglyoxal (AFB1 inducer) synthesis in AFLHPSi-1 cells by AREO and AREO-CsNe confirmed their novel antiaflatoxigenic mode of action. In-silico molecular docking studies revealed effective interaction of linalool with Ver-1 and Omt-A proteins, leading to inhibition of AFB1 biosynthesis. Further, AREO-CsNe showed enhanced antioxidant activity with IC50 values 3.792 and 1.706 µl/ml against DPPH• and ABTS•+ radicals, respectively. In addition, AREO-CsNe caused 100% protection of stored millets (Setaria italica seeds) from AFB1 contamination and lipid peroxidation over a period of 1 year without compromising its sensory properties and exhibited high safety profile with LD50 value 9538.742 µl/kg body weight. Based on enhanced performance of AREO-CsNe over AREO, it can be recommended as a novel substitute of synthetic preservative for preservation of stored millets.

Mechanistic investigations on antifungal and antiaflatoxigenic activities of chemically characterised Carum carvi L. essential oil against fungal infestation and aflatoxin contamination of herbal raw materials.

This study aimed to investigate the efficiency of chemically characterised Carum carvi essential oil (CcEO) against aflatoxin B1 (AFB1) producing strain of Aspergillus flavus (AF-LHP-WS-4) causing deterioration of herbal raw materials (HRM). GC-MS analysis of the EO revealed the presence of carvone (69.85%) as a dominant component. CcEO caused complete suppression of A. flavus growth and AFB1 secretion at 0.7 and 0.6 µL/mL, respectively. The investigation on antifungal mode of action showed that CcEO inhibited fungal growth via abrogating ergosterol biosynthesis and triggered efflux of vital cellular ions. The inhibition of AFB1 biosynthesis was attributed to the inhibition of cellular methylglyoxal (MG) biosynthesis. In addition, CcEO showed remarkable antioxidant activity (IC50 = 10.564 µL/mL) against DPPH (2,2-diphenyl-1-picrylhydrazyl) radicals. Based on overall results, it can be concluded that the CcEO may be recommended as potential antifungal agent for protection of HRM from fungal infestation and AFB1 contamination.