Assessing the preservative efficacy of nanoencapsulated mace essential oil against food borne molds, aflatoxin B1 contamination, and free radical generation
Introduction
Mycotoxin contamination is one of the serious problems of stored food commodities viz., fruits, vegetables, spices, legumes, cereals, and their shelved products. Ingestion of the mycotoxin contaminated food commodities may cause hazardous health effect to the human being and livestock. Currently, 300 different mycotoxins have been reported, among all aflatoxin B1 produced by Aspergillus flavus, A. parasiticus, A nomius has been received the considerable attention to the industries and regulatory authority in view of its carcinogenic, genotoxic, and immunosuppressive effect. Currently, a range of chemical preservatives viz., benzoic acid, sorbic acid, propionic acid, formic acid, and fumigants have been used to retard the molds growth. However, in view of green consumerism and suspected negative effect such as resistance and residual toxicity the industries are looking towards safer alternatives. Nowadays, the use of plant products having strong antimicrobial potential have been paid considerable attention to the industries as the preferred alternative to synthetic antimicrobial agent (Prakash et al., 2018; Hammer, Carson, & Riley, 1999; Morris, Khettry, & Seitz, 1979). Among plant product, essential oils (EOs) of aromatic plants often showed strong antimicrobial and antioxidant activity (Nychas, 1995:; Prakash et al., 2018). However, due to their volatile nature most of them fail to exert their antimicrobial and antioxidant activity in the food system (Prakash et al., 2018). Thus, encapsulation of essential oils using food grade coating materials may enhance their antimicrobial potency and successfully address the existing limitations in the food system (Donsì, Annunziata, Sessa, & Ferrari, 2011). Encapsulation is a process of packing and protecting of the bioactive parts such as polyphenols, enzyme, antioxidants, and nutraceuticals within a wall material with improved stability and controlled release in the food system (Gouin, 2004). In the past few years, chitosan has been widely used as the wall material of various plant-based preservative agents owing to its good-film forming capacity, biocompatibility, biodegradability and non-toxic nature (Donsì et al., 2011; Harris, Lecumberri, Mateos-Aparicio, Mengíbar, & Heras, 2011; Hosseini, Zandi, Rezaei, & Farahmandghavi, 2013; Luo, Zhang, Whent, Yu, & Wang, 2011; Muzzarelli, 2010).
Mace (dry Myristica fragrans Houtt. fruit covered with orange lacy aril) is widely used as culinary products and has potent antioxidant and antimicrobial activity (Kulandhaivel et al., 2011; Singh, Marimuthu, Murali, & Bawa, 2005). Although, the antifungal and antioxidant activity of Mace essential oil (MEO) has been reported previously, the mechanism of action against fungal growth and aflatoxin B1 suppression of crude MEO and its nanoformulation is lacking in the literature so far. The aim of the present investigation was to develop and characterize chitosan-cinnamic acid based nanogel as a carrier agent of mace EO. The nano-encapsulated mace essential oil (Ne-MEO) has been characterized through SEM, XRD and FTIR analysis. Further, the mode of action of Ne-MEO has been elucidated in term of its effect on the C- source utilization, ergosterol content, membrane integrity (ion leakage) and interaction of major bioactive component of MEO (myristicin) with ver-1 gene responsible for aflatoxin B1 production by A. flavus.
Section snippets
Chemicals and equipment
All the chemicals used in the present study were procured from Sigma Chemical Co. (St. Louis, MO, USA), Hi-media, and Sisco Research Lab (Mumbai, India). The equipments used were gas chromatograph mass spectrometer GC-MS (PerkinElmer Turbomass Gold, USA), UV transilluminator (Zenith Engineers, Agra, India), spectrophotometer (Shimadzu UV-1800), atomic absorption spectrophotometer (AAS) (Perkin Elmer Analyst 800), pH meter (Eutech Instruments, cyberscan), probe sonicator (Labman Scientific
Statistical analysis
The experiments were performed in triplicate and the data presented are the mean ± SE. The results were subject to by one-way analysis of variance and Tukey's multiple-range tests to identify significant differences in the comparison of means. P values of≤0.05 was considered significant. The analysis of present data was performed with SPSS program version 16.0 for Windows (SPSS Inc., IBM Corp.).
Mycobiota analysis and detection of the toxigenic strain of Aspergillus flavus
Mycoflora analysis revealed that the legumes seed samples were profoundly infested with molds species including toxigenic species of Aspergillus flavus. Moisture content and pH of the commodities ranged between (6.2%–11.3%) and (pH 5.4 - pH 6.3) respectively (Table 1). Highest moisture content (11.3%) was recorded with Vigna unguiculata (L.) Walp and the lowest (6.2%) for Cajanus cajan (L.) Millsp. The magnitude of pH was found in acidic range. A total of 11 fungal species were recorded from
Conclusion
Ne-MEO exhibited strong antifungal, antiaflatoxigenic and antioxidant activity over the free MEO; hence chitosan could be used as a carrier agent of plant-based antifungal and antioxidant agent. Further, the decrease in ergosterol content, leakage of vital cellular ions and impairment in C- source utilization in A. flavus (CP-05) exposed to Ne-MEO could be the possible mechanism of action against the growth and aflatoxin B1 production.
Conflicts of interest
The authors declare that there are no conflicts of interest.
Acknowledgment
The author gratefully acknowledges the financial support by UGC, New Delhi, India as Junior Research Fellowship (Ref: 370673). Bhanu Prakash and Akshay Kumar are thankful to Science and Engineering Research Board (Scheme No. ECR/2016/000299) New Delhi, India. We are also thankful to the Head, CAS in Botany, Banaras Hindu University, Varanasi for Instrumental facilities. We are also thankful to Indian Institute of Technology, Banaras Hindu University, Varanasi for SEM and XRD analysis.
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