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.

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.

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.

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.

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.

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.

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.

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.

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.