Green nanotechnology

Chapter 7

Green nanotechnology: nanoformulations against toxigenic fungi to limit mycotoxin production Velaphi C. Thipe1, 6, Pierce Bloebaum2, 6, Menka Khoobchandani2, 6, Alice Raphael Karikachery2, 6, Kavita K. Katti2, 6, Kattesh V. Katti2, 3, 4, 5, 6 1

Department of Chemistry, University of Missouri, Columbia, MO, United States; 2Department of Radiology, University of Missouri, One Hospital Drive, Columbia, MO, United States; 3 Department of Physics, University of Missouri, Columbia, MO, United States; 4Biological Engineering, University of Missouri, Columbia, MO, United States; 5Medical Pharmacology and Physiology, University of Missouri, One Hospital Drive, Columbia, MO, United States; 6 Institute of Green Nanotechnology, University of Missouri, One Hospital Drive, Columbia, MO, United States

1. Introduction Fungi are the most problematic microorganisms that cause significant agricultural losses due to their mycotoxigenic ability (produce mycotoxins-toxic secondary fungal metabolites) that frequently contaminate various agricultural products (e.g., cereals, peanuts, soybeans, sorghum, maize, and their by-products) [1e3]. Fungi have the ability to proliferate in a wide range of temperatures and pH and thus contaminate food and animal feed. Kaur et al. [1] reported that the chemical and thermal stability of mycotoxins during food processing (boiling, cooking, and pasteurization) makes it difficult to control. Mycotoxigenic fungi pose challenges to the agricultural sector through the decline in crop yield and the deterioration of harvest quality due to the presence of mycotoxins that affect human and animal health [4]. The consumption of mycotoxin-contaminated foods and feeds can result in acute or chronic toxicity in humans and animals. Additionally, this results in diseases that include actinomycosis, aspergillosis, candidiasis, mycoses, mycotoxicoses, otomycosis, and penicillinosis. Moreover, there is also a growing concern of consuming mycotoxins from animal-derived food products (meat, milk, and eggs) that contain residues/metabolites of mycotoxins [2]. Mycotoxigenic members of the Nanomycotoxicology. https://doi.org/10.1016/B978-0-12-817998-7.00007-0 Copyright © 2020 Elsevier Inc. All rights reserved.

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fungi genera Aspergillus, Fusarium, and Penicillium produce a wide array of mycotoxins which include aflatoxins (AFs), fumonisins, ochratoxins (OT), zearalenone (ZEA), and trichothecenes including deoxynivalenol (DON), nivalenol (NIV), and T-2 toxin; over 400 mycotoxins have been identified and reported. The prevalence of mycotoxins hinders international trading because approximately 25% of harvested crops worldwide is contaminated by mycotoxins annually [2]. This results in huge agricultural losses of countries, which affects the economy significantly. The United States’ corn industry suffered significant losses due to AF contamination from $52.1 million to $1.68 billion dollars. Mycotoxins in animal feeds significantly cause economic losses in the livestock industry due to stunted growth, reduced immunity to infections, decreased production of eggs, meat, and milk, liver and kidney damage, poor feed conversion, and increased mortality [5e7]. Although there is a variety of commercial synthetic antifungal agents, their limitation is governed by their toxicity and the onset of multidrug resistance strains; therefore, alternative ecofriendly, effective agents are continuously probed. The occurrence of mycotoxins has a wide distribution within the food chain. Most of the human diet is composed of one or more of the following staple foodsdcereals (maize [corn], millet, rice, sorghum, soybeans, and wheat), roots and tubers (cassava, plantain, potatoes, taro, and yams), and animal products such as eggs, cheese, meat, and milk [8,9]. Yuan et al. [10] reported the variation of mycotoxigenic fungi of trichothecenes and AFs in wheat grains during storage in various producers of wheat (China, Italy, Iran, Argentina, Kazakhstan, and Saudi Arabia) predominantly manifested by Fusarium graminearum (F. graminearum), DON, NIV, and Aspergillus flavus (A. flavus), AFs [7]. Staple foods are mainly susceptible to mycotoxin contamination; thus, mycotoxin contamination in food and feed is a global issue and has attracted attention from many national and international public health and governmental authorities [10]. These authorities include, but not limited to, the Food Agriculture Organization, World Health Organization (WHO), European Food Safety Authority, US Food and Drug Administration (FDA), and the Joint Expert Committee on Food Additives. This chapter explores the antifungicidal properties of phytoextracts and phytochemicals from plant against fungi. We further summarize the potential application of Green Nanotechnology for the production of nanoformulations using phytochemicals derived from staple foods and plant materials for their effectiveness against mycotoxigenic fungi and subsequent reduction in mycotoxin production.

2. Major mycotoxins This section briefly discusses the major mycotoxins, their occurrence, and toxicity toward human and animal health. The contamination of food and feed

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stuff products by mycotoxins can occur at several stages during the food chain cycledthis includes during pre- and postharvest, packaging, distribution, and storage [9]. Scientists, researchers, and representatives from industry, government, and academia focus on the importance of addressing concerns about all aspects of mycotoxin research during the International Symposium on Mycotoxins and Toxigenic Fungi (MYTOX) meetings. A report by Saeger et al. [11] highlighted the urgency for multidisciplinary efforts to alleviate mycotoxin contamination within the food chain, from the field to consumerda “One Health” concept [11].

2.1 Aflatoxins AFs are the most toxic mycotoxins produced mainly by A. flavus and Aspergillus parasiticus causing aflatoxicosis. A. flavus produces aflatoxin B1 (AFB1) and B2 (AFB2), whereas A. parasiticus produces AFB1, AFB2, G1 (AFG1), and G2 (AFG2) that contaminates staple foods as eluded previously. Aflatoxigenic fungi can proliferate in a variety of foods such as cereals (maize [corn], millet, rice, sorghum, soybeans, and wheat) and nuts (almonds, cottonseeds, groundnuts, peanuts, pistachio nuts, and walnuts). Aflatoxin M1 (AFM1) and AFM2 contaminate dairy products such as milk and cheese due to ingestion of contaminated feed with AFB1 and AFB2 by animals which are translated by hepatic microsomal cytochrome P450, respectively [2,12]. The AFs have carcinogenic, hepatotoxic, immunosuppressive, mutagenic, and teratogenic properties with the liver being the target organ. Among the AFs, AFB1 is classified as a Group 1 carcinogen according to the International Agency of Research on Cancer (IARC) and the most toxic of all AFs because it has been reported to cause hepatocellular carcinoma (Fig. 7.1) [9,13]. The cytotoxic effect of AFB1 is due to the binding of bioactivated AFB18,9-epoxide to cellular macromolecules (i.e., mitochondrial, nuclear nucleic acids, and nucleoproteins), resulting in general cytotoxic effects [12]. Several studies have reported that exposure to AFs is associated with stunted growth in children and immune system disorders.

2.2 Ochratoxins OTs are secondary metabolites produced by members of Aspergillus and Penicillium genera. Ochratoxin A (OTA), B (OTB), and C (OTC) pose health

FIGURE 7.1 Chemical structure of aflatoxin B1.

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FIGURE 7.2 Chemical structure of ochratoxin A.

problems in humans and animals as shown in Fig. 7.2 [14]. The most important OT is ochratoxin A (OTA) and is produced mainly by Aspergillus ochraceus, Penicillium verrucosum, and some species of Penicillium. OTA is classified as a Group 2B carcinogen according to the WHO and IARC responsible for affecting human and animal health [13,15,16]. Lappa et al. [14] described OTA as a hepatotoxic, immunosuppressive, nephrotoxic, mutagenic, teratogenic, and carcinogenic agent. It has been confirmed that OTA accumulates in the kidney and is considered to have a major role in Balkan endemic nephropathy [15]. There had been some contrivances about the cytotoxic mechanism of OTA reported by researchers and the reported data are contradictory. A consensus was reached that suggests that OTA unlikely acts through a single pathway to achieve its toxicity [16].

2.3 Fumonisins Fumonisins are produced by toxigenic Fusarium spp. but mainly produced by Fusarium verticillioides and Fusarium proliferatum [17]. Fumonisins are composed of fumonisin B1 (FB1), fumonisin B2 (FB2), and fumonisin B3 (FB3), where FB1 is the most toxic which is associated with esophageal cancer (Fig. 7.3) [9]. FB1 has been classified as a Group 2B carcinogen by the IARC due to its toxicity. Eluded previously, maize is one of the staple foods, the third

FIGURE 7.3 Chemical structure of fumonisin B1.

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most important cultivated crop after wheat and rice and is susceptible to the proliferation of F. verticillioides and F. proliferatum that are responsible for the production of fumonisins [13]. Bryta et al. [18] described the effects of the consumption of fumonisins-contaminated food and feed induces leukoencephalomalacia in horses, pulmonary edema in swine, liver injury, nephrotoxicity, liver cancer in rats, arteriosclerosis in primates other than humans, and esophageal cancer in humans [17].

2.4 Zearalenone The mycotoxin ZEA is a secondary metabolite produced by toxigenic Fusarium spp. and predominantly frequent in contaminated animal feed. Fertility and reproductive toxicity by ZEA are attributed by ZEA being an estrogenic toxin that has a similar structure with 17 b-estradiol for estrogen receptor binding (Fig. 7.4) [7]. ZEA is an endocrine disruptor that affects normal steroid hormone synthesis (estradiol, progesterone, and testosterone) [19]. Chronic exposure to ZEA can lead to hyperestrogenic syndromes because of its immuno-, hepato-, and genotoxicity [20].

2.5 Trichothecenes Trichothecenes mycotoxins are secondary metabolites produced by Fusarium spp. There are two types of trichothecenes that cause significant public health concerns due to their toxicity and occurrence (Type A and B). Type A group includes T-2 toxin (T-2), HT-2 toxin (HT-2), neosolaniol (NEO), and diacetoxyscirpenol (DAS) and Type B group includes DON, NIV, and 3- and 15-acetyldeoxynivalenol (15-ADON) as shown in Table 7.1 [6]. The general structure of trichothecenes is shown in Fig. 7.5. T-2 and DON are the most prevalent and toxic of the trichothecenes and T2 can be metabolized into HT-2 [21]. Type A trichothecenes are produced mainly by Fusarium sporotrichioides and Fusarium poae, whereas Type B trichothecenes are mainly produced by Fusarium graminearum and Fusarium culmorum [22]. DON and T-2 are immunosuppressive agents that induce apoptosis and inhibit protein synthesis by disrupting the functioning of ribosomal subunits and brain neurochemical changes [19,23].

FIGURE 7.4 Chemical structure of zearalenone.

Type

Trichothecenes

Abbr.

R1

R2

R3

R4

R5

A

T-2 toxin

T-2

OH

OAc

OAc

H

OCOCH2CH(CH3)2

HT-2 toxin

HT-2

OH

OH

OAc

H

OCOCH2CH(CH3)2

Neosolaniol

NEO

OH

OAc

OAc

H

OH

Diacetoxyscirpenol

DAS

OH

OAc

OAc

H

H

Deoxynivalenol

DON

OH

H

OH

OH

¼O

Nivalenol

NIV

OH

OH

OH

OH

¼O

3-Acetyldeoxynivalenol

3-ADON

OAc

H

OH

OH

¼O

15-Acetyldeoxynivalenol

15-ADON

OH

H

OAc

OH

¼O

B

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TABLE 7.1 Basic chemical structure of trichothecenes.

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FIGURE 7.5 Chemical structure of trichothecenes.

T-2 and HT-2 have been reported to potentially induce anorexia and impair immune function by targeting the appetite center. This inhibits DNA, RNA, and protein synthesis and leads to hemorrhaging and neurological disorders; this was confirmed through in vivo analysis in various animal models such as mice, rodents, cats, pigs, Holstein cows, and broiler chickens [7,22,24e26]. A study by Zhang et al. [21] authenticated that Type A trichothecenes T-2, HT-2, NEO, and DAS induced a dose-dependent anorectic response in mice via oral and intraperitoneal injection (IP) administration. The anorectic response was ranked based on the mode of administration, where for oral exposure T-2, HT-2, and NEO > DAS and IP administration DAS > T-2, HT-2, and NEO.

3. Medicinal plants This section discusses the use of plant-derived products as such phytofungicides that are being used to reduce the proliferation of mycotoxigenic fungi with a subsequent decrease in mycotoxin production.

3.1 Phytochemicals against mycotoxigenic fungi Medicinal plants have been used for centuries to treat various diseases; the effectiveness of the treatment is attributed by the composition of bioactive compounds present in the plant that includes phenylpropanoids, quinones, terpenoids, steroids, alkaloids, and their derivatives. Phytochemicals is a term generally used for various nonnutritive biologically active chemical compounds of plant origin (Fig. 7.6) [27]. A number of studies have been investigated in the use of bioactive compounds isolated from medicinal plants against different mycotoxigenic fungi. Maize and its derivatives are good sources of various nutritional compounds which include starch, proteins, lipids, and different bioactive compounds that can be used as antifungal agents [8]. Phytofungicides are formulations made by extracts from different parts of the plants (leaves, bark, stem, and roots), which have a different distribution of the phytochemicals. Essential oils and plant extracts can be derived from cinnamon, clove, garlic, ginger, green tea, ground red pepper, spices, herbs, potato, mint, neem leaf and oil, onion bulb, olive oil, oregano, grapefruit peel, lemon, orange, etc. Razzaghi-Abyaneh et al. [28] evaluated the efficacy of

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FIGURE 7.6 Major phytochemicals from various plants.

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neem leaf and seed extracts against A. parasiticus which inhibited fungal mycelia growth by 90%. Recently, Reddy et al. [29] evaluated the efficacy of antifungal component in clove plant extracts on mycelial growth of A. flavus, A. parasiticus, Aspergillus niger, and Aspergillus ochraceus. There is a considerable interest to develop effective fungicidal modalities against toxigenic fungi with subsequent low/no mycotoxin production without affecting the growth and productivity of the staple foods and feeds. Essential oils extracted from anise, basil, caraway, chamomile, cinnamon, fennel, marigold, spearmint, thyme, and alyssum were tested against A. flavus, A. parasiticus, A. ochraceus, and Fusarium moniliforme. The data reported by Soliman and Badeaa [30] revealed that essential oils from anise, cinnamon, spearmint, and thyme at 500 ppm were more effective against the tested fungi and reduced mycotoxin production in wheat grains treated with the oils. Anjorin et al. [27] mentioned that essential oils extracted from clove have shown some antifungal activities. The use of bioactive compounds for antifungal applications has proved to be a beneficial approach for the alleviation of contamination caused by mycotoxigenic fungi with subsequent mycotoxin reduction. The high level of natural antioxidants found in plant material is useful for a variety of applications that stem from cancer therapy, biofuel, and recently in the agricultural sector. The bioactive components and essential oils extracted from plants consist of numerous antioxidants, as shown in Table 7.2. The major bioactive components are phenolic compounds, which are common secondary metabolites in plants that have health benefits facilitated by their rich-electron density. The properties of phytochemicals (anticarcinogenic, antiinflammatory, antibacterial, and antifungal) make them promising ingredients in the development of ecofriendly products that can be utilized in the food industry and for health applications [31]. Shehata et al. [31] studied the use of oil-bioactive films from agricultural wastes (immature fig fruit (ImFF), fig leaves (FL), and pomegranate husks (POH)) for their antifungal properties contra toxigenic fungi. Their approach used jojoba oil (gadoleic C20:1 fatty acid), which a liquid extracted from the Simmondsia chinensis plant seeds, as a carrier for coating soybean grains with ImFF, FL, and POH. The results obtained showed that ImFF extract had low toxicity against brine shrimp but the highest antifungal efficacy against all tested mycotoxigenic strains (A. parasiticus, A. fumigates, P. chrysogenum, and P. expansum) demonstrated that the formulation was selective for the fungi strains. This was attributed by the high total phenolic and flavonoid content in the ImFF extract compared with the FL and POH extracts. Moreover, the ImFF degraded AFs (AFB1, AFB2, AFG1, and AFG2) by 52.7%. The jojoba oil-ImFF extract film protected soybean grains against fungal contamination and mycotoxin production. A study by Liao et al. [32] reported the beneficial effects dietary supplementation of L-arginine (Arg) on piglets fed DON-contaminated feed.

Databases Database

Description

Website

Dr. Duke’s Phytochemical and Ethnobotanical Databases

Facilitates an in-depth understanding of the plant, chemical, specific activity, and ethnobotany searches by using either scientific or common names.

https://phytochem.nal.usda.gov/phytochem/search

USDA Phytochemical & Ethnobotanical Databases

Extensive information about chemical single plant synergy (specific activities), toxicity, and chemical composition.

http://www.leffingwell.com/plants.htm

Phenol-Explorer

Comprehensive database about natural phenols and polyphenols in food.

http://phenol-explorer.eu/links

PhytoHub

Detailed information about phytochemicals and their human metabolites.

http://phytohub.eu/

Phytochemica

A platform to explore phytochemicals of medicinal plants.

http://faculty.iiitd.ac.in/wbagler/webservers/Phytochemica/

Chemical Entities of Biological Interest (ChEBI)

A dictionary focused on small bioactive chemical compounds.

https://www.ebi.ac.uk/chebi/

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TABLE 7.2 Medicinal plants and respective phytochemical databases.

A phytochemical database that contains data approximately 16,332 chemical compounds including quantitative data were available, localization and their specific activities.

Beckstrom-Sternberg et al. [67]

Natural Medicines Comprehensive Database

Database for effectiveness and drug interaction of natural products on various diseases.

http://naturaldatabase.therapeuticresearch.com/home.aspx? cs¼&s¼ND&AspxAutoDetectCookieSupport¼1

Herb Research Foundation

Detailed information about herbs.

http://www.herbs.org/herbnews/

American Herbal Products Association

Advocates for Pharmacopeia for effective laws and regulations that promote the responsible commerce of herbal products.

http://www.ahpa.org/

American Herbalists Guild: An association of herbal practitioners

Promotes clinical herbalism as a viable profession for translational medicine rooted in ethics, competency, diversity, and freedom of practice.

https://www.americanherbalistsguild.com/

A platform to educate the public about herbal medicine as an accepted part of healthcare.

http://abc.herbalgram.org/site/PageServer

Subscription-based American Botanical Council

Continued

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Databases Database

Description

Website

Medical Herbalism: A Journal for the Clinical Practitioner

A journal that strengthens the herbal practitioner to work with medical doctors for preserving and developing the science and understanding of herbal medicine. Thereby, promoting communication and sharing of clinical methods and experiences.

http://www.medherb.com/MHHOME.SHTML

Natural Product Reports

A review journal that stimulates progress in all areas of natural products research.

http://pubs.rsc.org/en/journals/journalissues/np#! recentarticles&adv

Journals

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TABLE 7.2 Medicinal plants and respective phytochemical databases.dcont’d

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The results revealed that Arg exerted a protective role in pigs fed DONcontaminated diets that was facilitated by the ability of Arg to stimulate phagocyte activity thereby accelerating the elimination of endotoxin in the gut. There has been a major focus in the development of new antifungal drugs from natural sources as alternatives to synthetic chemicals [27]. Several studies have reported on the inhibitory effect of various plant extracts [27]. Clove is rich in eugenol and been reported to be useful in controlling mycotoxigenic fungi and mycotoxins [27]. Kumar et al. [33] studied the efficacy of Holy Basil (Ocimum sanctum) which is herb used within Ayurvedic medicine against the fungi responsible for the biodeterioration of food during storage. Results demonstrated that the essential oils extracted from O. sanctum (eugenol being the major component) exerted an inhibitory effect at 0.2 mL/mL against A. flavus. Moreover, this significantly inhibited AFB1 production. This was supported by results reported by Reddy et al. [34] who demonstrated that rice grains treated with 2.4 mg eugenol per g of grains inhibited A. flavus growth and reduced AFB1 biosynthesis. A study by Dambolena et al. [35] reported the effectiveness of the antifungal activity of essential oils extracted from Ocimum basilicum L. and Ocimum gratissimum L. against F. verticillioides, which reduced the production of fumonisins. Main functional groups are composed of phenol and carboxyl that are involved in some different adsorption mechanisms [36]. The high surface to volume ratio of the nanoparticles can facilitate the increased adsorption of mycotoxins. Medicinal plants have been widely used worldwide, estimated 80% herbal remedies and 25%e50% raw materials in the pharmaceutical industries for the prevention and treatment of disorders and diseases [13]. However, the use of medicinal plants has its own limitations and is not exempted from contamination by various fungi. AFs, FBs, OTA, and ZEA contamination in medicinal plants has been reported from various countries [37]. On the basis of a review by Ashiq et al. [13], plant materials destined for medicinal uses should be carefully evaluated before being used in formulations in the development of drugs. Plant-based products, especially the bioactive phytochemicals, are recognized as one of the most promising alternative routes for the development of effective and safe antifungal agents.

3.2 Challenges of phytofungicides The use of phytochemicals as antifungal agents is hindered by several factors which primarily include availability of plants materials, low bioactive phytochemical extraction (which impacts on the quality of the product), reproducibility, lack of phytopesticide analysis facilities, and lack of infrastructure for upscale production [27]. Anjorin et al. [27] stated that the transfer of knowledge from herbalists/traditional healers to academia to industries and government is limited by the lack of fostering a cohesive research quality network as shown in Fig. 7.7 [38].

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FIGURE 7.7 Knowledge transfer framework for research quality in modern medicine.

The phytochemicals that exhibit inhibitory effects on most toxigenic fungi which will be enhanced to prevent photodegradation and increase the shelf life of the compounds. The combination of the two or more bioactive phytochemicals in a formulation could increase the fungicidal spectrum against the toxigenic fungi. Anjorin et al. [38] demonstrated the combination of two different plant extracts was more effective than the plant extracts used separately and reduces the development of resistance by toxigenic fungi further supported these results. In recent years, a number of organizations have developed great tools such as phytochemical databases that have collective information about the plant material, phytochemical composition, major bioactive compounds, structure, and properties of the phytochemical as shown in Table 7.2. However, there are no databases that provide information about plant materials and bioactive phytochemical with antifungal potential against toxigenic fungi. As mentioned above, the lack of a knowledge transfer framework limits the translation of phytofungicides in agricultural practices. Establishing a database for a shared knowledge transfer from indigenous herbalists about promising plants that require further studies by academia and industries would vastly improve the development of antifungal agents. This knowledge and data are of profound importance in pharmaceutical, nutritional, and biomedical research toward alternative therapies and herbal products.

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A number of chemical commercial fungicides affect the integrity of the food and feed, thereby compromising the health and safety of human and animals. This has led to legal acts and regulations that govern the pest management requirements to limit the use of harmful fungicides. Another disadvantage of organic synthetic fungicides besides high toxicity is the propensity of residuals remaining in the food and feed after treatment. Thus, new strategies that are eco-friendly, economically feasible, and present no toxic effects on human and animal health are required. In organic farms, chemical substances used to limit fungi manifestation with subsequent mycotoxin production are prohibited; therefore, a new generation of fungicides that capitalizes the bioactive compounds present in plants with the integration of green nanotechnological approaches can be beneficial [22].

4. Green chemistry principles Nanotechnology has revolutionized a number of disciplines, and nanomaterials have attracted tremendous attention due to their properties. In the agricultural sector, the main focus is on pathogen detection using highly selective and sensitive biosensors and development of novel antimicrobial agents. Phytochemicals have been reported to have antimicrobial, antiinflammatory, antimodulatory, and anticancer properties. The high surface to volume ratio allows for a high payload of phytochemicals that provides a strong antimicrobial effect. Metallic and polymeric nanoparticles have the benefit of ease in production. Metallic nanoparticles that have been reported to be effective against toxigenic fungi include gold, silver, copper, iron, and zinc nanoparticles and/or alloys of these metals. Multidrug-resistant strains are problematic in all sectors (agriculture and medicine), therefore multiple modes of action are required to deal with thisdsilver nanoparticles (AgNPs) have been reported to possess multiple modes of inhibitory action against a microorganism [1]. There are a number of suggested mechanisms about the mode of action for antifungal activity of the synergistic effects of the nanoformulations. The mode of action includes the formation of reactive oxygen species (ROS) that disrupt the cell membrane integrity assisted by the activity of the attached phytochemicals.

4.1 Metallic nanofungicides In this section, we discuss the application of green nanotechnological solutions for migrating fungal growth and reducing the occurrence of mycotoxin contamination through the use of nanomaterials. The general strategies to inhibit and eliminate the growth of mycotoxins are discussed below with the utility of metallic nanoparticles. It has been observed from several years of research that metallic nanoparticles are extensively used as antimicrobial agents for antibiotic resistance of pathogenic bacteria and fungi [39e54].

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Bacteria are prokaryotic and fungi are eukaryotic organisms, which make them possess unique characteristics; therefore, it requires effective drug moieties tailored for fungi, which could inhibit fungal growth and resurgence of mycotoxins. Sierra et al. [55] provided evidence that among metallic nanomaterials, zinc and silver oxide are effective against mycotoxins. Metal oxide nanoparticles are characterized by high surface area to volume ratio. Inorganic compounds such as Zn have attracted interest due to their nontoxicity at low concentrations that present strong antimicrobial activity. The advantage of Zn is that it serves as an essential element utilized in dietary supplements. Moreover, the FDA considers ZnO (21 CFR 182.8991) as generally recognized as a safe (GRAS) substance [56]. Savi et al. [23] described that Zn is essential for several physiological and metabolic pathways especially the immune system. Moreover, Zn is also an essential micronutrient for plants that is supplemented in fertilizers. Savi et al. [23] demonstrated the use of zinc compounds (ZnO-NPs, ZnO, ZnSO4, Zn(ClO4)) as antifungal and antimycotoxin agents against A. flavus, F. graminearum, and P. citrinum. The results revealed that ZnSO4 and Zn(ClO4)2 treatments at 100 mM exhibited stronger antifungal and antimycotoxin activities compared with ZnO-NPs and ZnO. Moreover, an increase in the production of ROS in the fungi hyphae treated with Zn compounds was observed, thus affecting the fungal cellular metabolism. Zinc salt has been used for the treatment of zinc deficiency. De Romana et al. [55] deliberated the influence of ZnO nanoparticles on Botrytis cinerea and P. expansum. ZnONP produces ROS, which led to the inhibition of the bilayer of the cell membrane and induces cell death [55]. The antifungal effect of ZnO-NPs against the growth of mycotoxigenic fungi and mycotoxins production in feed and food has been evaluated [57]. The authors Sawai and Yoshikawa [57] discussed antifungal and antibacterial efficacies of metal NPs (ZnO, MgO, and CaO) against Staphylococcus aureus, E. coli, and fungi. Hernandez-Melendez et al. [56] produced flower-shaped ZnO nanostructures (ZnONs) via aqueous precipitation route at room temperature. The antifungal and antiaflatoxigenic activity properties of the resultant nanostructures was investigated against A. flavus. ZnONs morphology demonstrated as structurally arranged flattened tipped hexagonal nanorods at 120 nm in size. The antifungal and antiaflatoxigenic activity of ZnONs was observed at 1.25 mM for mycelial reduction up to 78% and a 98.7% AF content decrease, respectively. Maize treated with 100 mg ZnONs per g of maize were protected from fungal proliferation with relatively low AF production. The nanoparticle’s shape affects various conditions such as binding affinity to a cell’s surface membrane, so one can expect that the shape of the nanoparticle also contributes to its antifungal effects [58]. Barua et al. [59] showed that oblate, rodlike nanoparticles increase the inhibition of eukaryotic cell growth five times as much as nanospheres of the same dose.

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Among metallic nanoparticles, AgNPs have received serious attention as an antifungicidal agent. AgNPs inhibit fungal growth and metabolic changes to inhibit mycotoxins production [60,61]. The application of 45 ppm AgNPs was reported to inhibit mycotoxin production (up to 80%), and changes in the enzymatic profile in A. niger and P. chrysogenum [60]. In another study, the effects of AgNPs on growth and AFB1 production of AF-producing A. parasiticus were reported at concentration 90 ppm [62]. Work by Deabes et al. [63] evaluated the effectiveness of AgNPs synthesized by fungi (A. terreusHA1N (KR364880) and P. expansum HA2N (KR269857)) against toxigenic A. flavus on the production of AFB1. The data obtained revealed that the synthesized AgNPs-HA2N and AgNPs-HA1N reduced the proliferation of A. flavus with the diminution of AFB1 production. This was found to be attributed by the effect of AgNPs on omt-A enclosed in the AF biosynthetic pathway that regulates AF aflR gene production [63]. Abdel-Hadi et al. [64] synthesized AgNPs using A. terreus MALEX and investigated the biological activity against mycotoxigenic fungi (A. flavus, A. ochraceus, F. solani, and Alternaria alternate). F. solani and A. alternate were more susceptible to the treatment at 20 mM AgNPs. The AgNPs may be useful to control AF contamination of susceptible crops in the field. The high phytochemical payload anchoring on the surface of the nanoparticles increases the efficacy of the antifungal activity, thereby inducing fungal death as shown in Fig. 7.8. Mofilikoane [65] investigated the use of copper nanoparticles (CuNPs) as enhancers for the improvement of conventional antibiotics. The reported data revealed that the incorporation of CuNPs increased the antifungal activity of selected antimicrobial drugs (Amphotericin B, Clotrimazole, Nystatin, Fluconazole, and Ciprofloxacin) by 1.6 folds. Another study by Savi et al. [66] investigated the zeolite derivatives against A. flavus and AFB1 as an antifungal and absorbent material, respectively. Ion-exchanged zeolite, a microporous crystalline hydrated aluminosilicates composed of tetrahedral linked [SiO4]4 and [AlO4]5 through a shared O2 atom, has been reported to have excellent absorbent and catalytic properties used as carriers support matrix for antifungal agents and as a mycotoxin binder for decontamination. High antifungal activity against A. flavus was observed using ion-exchanged zeolites with Cu2þ and Liþ at 2 mg/mL. Furthermore, zeolite-Cu2þ and zeolite-Liþ significantly inhibited conidia germination, AFB1 production, and zeolite-Li þ showed superior AFB1 adsorption capacity at 100% in all concentrations. Gold nanoparticles (AuNPs) are mainly used as a screening of mycotoxins [62]. However, some of colloidally stabilized AuNPs having sizes in the range of 3e20 nm represented that antifungal effects were investigated for Penicillium [67]. The authors Eid et al. [62] discussed and provided strong evidence that the consideration of AuNPs as antifungal material could evade the passive immune effects of other antifungal material along with the ability of targeting specific sites [67]. Carbon nanomaterials are one of excellent stable NPs with high adsorptive properties and large surface are per weight for their application for mycotoxin elimination.

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FIGURE 7.8 Antifungal activity of phytochemical-conjugated nanoparticles.

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4.2 Polymeric nanofungicides Most of the polymeric nanoparticle research in literature focuses on antibacterial applications rather than antifungal. Several synthetic and natural polymers have been studied as potential candidates for the production of nanoparticles as vehicles for antifungal actives in agriculture, plant-based applications, biomedical applications, and food packing industry. Past findings in the field of antifungal nanoparticles are dominated by chitosan (CS) or their derivatives [47,68,69]. CS is a linear polysaccharide (D-glucosamine and N-acetyl-D-glucosamine) and has been demonstrated to have antifungal activity against various plant pathogens. CS’s mode of action involves the disruption of the fungal cell membrane [47]. The source of polymers used as a support matrix for nanoparticles can be classified into the following: (1) synthetic and (2) natural polymers (also known as biopolymers). Some studies use synthetic polymers (e.g., PEG) as a scaffold for biodegradable and plant-based bioactive preparation [70]. Other studies use naturally occurring compounds or biopolymers to enhance the antifungal effect of fungicides. Of these, CS is among the most organismfriendly and cost-effective [71]. Polymeric nanoparticles are further classified based on their role of combating fungal proliferation and mycotoxin contamination: l

l

Functionaldthis involves the production of nanoparticles with polymers like CS that are inherently antifungal in nature. This enhances the efficacy of the encapsulated antifungal agent. Structuraldpolymeric nanoparticles release sustained and therapeutic amounts of bioactive phytochemicals. The bioactive compound encapsulated in a polymeric nanocage will be released under the appropriate physiological conditions (e.g., the presence of enzymes, higher temperature, and pH change) [71] as shown in Fig. 7.9. The structural polymeric nanoparticle can be further classified based on preparation and architecture, into nanospheres, nanocapsules, and polymeric micelles [70].

CS is the most popular biopolymer, and there are multiple CS-based polymeric particles in the literature for antifungal applications. CS nanoparticles (CSNPs) were effective at inhibiting fungal proliferation and their role has been studied in decreasing the pathogenicity of infection [72]. Other biopolymers like polylactic acid (PLA) synthesized from bioderived monomers can be applied as a film for antimicrobial food packaging. Another study used poly(lacticco-glycolic acid) (PLGA), PLA, and alginate (Alg) nanoparticles to efficiently encapsulate antifungal drug Nystatin and improve its interaction with mucin and human colonic cell line [67]. These formulations were promising in terms of their mucoadhesive capacity while being nontoxic/nonirritating at the same time [72]. Biodegradable PLGA nanoparticles have been studied as a potential nanodelivery system for the food antifungal compound natamycin [73].

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FIGURE 7.9 Agricultural application of resveratrol-loaded chitosan nanoparticles (Res-CSNPs). (A) In-field crop plantation, (B) unhealthy AFB1 contaminated corn, (C) healthy decontaminated corn, and (D) the synergistic mechanism of Res-CSNPs, where the antifungal activity of resveratrol mitigates fungal proliferation that subsequently reduces mycotoxin production and the high adsorption capacity of chitosan allows for mycotoxin decontamination by remediation.

Spirulina seems to have polyhydroxyalkanoates, a family of biopolymers predominantly extracted from cyanobacteria, including spirulina, and used to provide non-toxic biocompatible scaffolding for biomedical applications [74]. Antifungal polymeric nanoparticles were synthesized through ring opening polymerization and atom-transfer radical polymerization reactions of poly(epsilon-caprolactone) (PCL) and poly(N,N-dimethylamino-2-ethyl methacrylate) (PDMAEMA), or methoxy polyethylene glycol (PEG). These polymeric nanoparticles incorporating Amphotericin B (AmB) showed improved antifungal activity against Candida albicans, whereas polymeric nanoparticles themselves were not effective [75]. Hydrophobic antifungal drug Itraconazole incorporated in lipomers which are polymeric lipid hybrid nanoparticles improved intestinal permeability [76]. In addition to the biopolymeric and synthetic approaches, the antifungal strategy can include one or more of the following directions: l

One can alter synthetic experimental conditions: polymer length and charge of the polymeric nanoparticles, to change properties like chemical composition, surface charge, size, and drug release profiles by manipulating experimental conditions and polymer length and charges, e.g., food packaging [70].

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l

l

l

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Polymeric nanoparticles can be made functionalized for desired drugs and target ligands via surface conjugation [71]. Use polymeric compounds for the formation of hybrid antifungal nanostructures [76]. Biopolymer capping has been studied for enhanced biocompatibility of nanoplatforms. One study used it for maintaining entrapped curcumin [77]. A combination approach using biopolymer and synthetic chemistry.

In another study, a combination of CS and AgNPs was evaluated for synergistic antifungal activity against Pyricularia oryzae extracted from blastinfected leaves [78]. Kaushik et al. reported the amphiphilic polymer thiram formulation and that the fungicidal effect of thiram is responsible for limiting moisture, thereby reducing the rate of seed deterioration. The encapsulation and administration of bioactive plant extracts (phytochemicals) can be facilitated using polymeric nanoparticles to provide targeted drug delivery and to solve stability-related problems [79]. Moreover, some polymer-based nanoparticle system enables the use of smaller quantities of a fungicide such as flavonoid compounds and therefore offers a more environmentally friendly method of controlling fungal pathogens in agriculture. For example, lecithin/ CS nanoparticles have been used to overcome problems such as poor dissolution and bioavailability for flavonoid compounds that are strong antioxidant and antifungal agents [80]. Fouda et al. [81] reported a superior fungicides (biopolymer composite incorporated with k-carrageenan) showed higher activity against pathogenic fungi when compared with fluconazole.

4.3 Hybrid nanofungicides Hybrid nanofungicides are a unique subgroup of nanoparticles which are composed of two types of nanofungicides. Typically, these are metallic nanospheres encapsulated or doped into a polymer matrix [82e85]. Kaur et al. [1] showed that Ag/CS nanoformulation displayed superior antifungal activity as compared with the sole treatment of AgNPs and CS-NPs against tested fungi. Alone, polymeric and metallic nanoparticles demonstrate toxicity to microbes that are harmful to plants, so using these hybridized particles together amplify the antifungal properties these particles normally exhibit [33,84e86]. Among the various precursors used for hybrid particles today, the most common include silver, zinc, and copper (for metals), and CS polypropylene, silicon, copolyester, polyethylene, and polyether sulfone for polymers [82e84,87]. With all the different combinations of substituents possible, each outcome provides unique benefits supplementary to fungicidal activity. The key to revolutionizing this approach is to synthesize the particles in a green method, leaving the environment unharmed. Ag/CS-NPs are a candidate for widespread fungicide use because their synthesis is exactly this, green; these particles come in the form of having Ag both doped into the CS matrix

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and internalized in the CS matrix sphere [1]. In fact, Ag/CS nanoformulations can display superior antifungal activity as compared with the sole treatment of AgNPs and CS-NPs against tested fungi [1]. When tested against three common fungicides toxic to humans, the Ag/CS-NPs provided a zone of inhibiting microbial growth nearly twice the size of that of AgNPs and CS-NPs alone [1]. Aside from CS, other hybrid Ag/polymer-NPs synthesized by green methods not only are environmentally safe but more antimicrobial and less cytotoxic than AgNPs alone [88]. As shown in Fig. 7.10, they are many modes of antifungal activity that can be mediated by light activation, increased antifungal activity by integrating other moieties, and decontamination of mycotoxin. Hybrid ZnO-NPs were effective in preventing mold and fungal growth after being exposed to a fluorescent light source [89]. Adjusting the heat levels on thermoresponsive polymer coatings shows new ways to exploit the engineering of materials that require fungicides in various environments [88]. Use of current pesticides and fungicides are causing many health concerns around the world. The potential impact these nanofungicides have on humans after use must be considered. One of the ways to assess the threat to human health rests in the release of the antimicrobial growth agent. Preliminary studies of hybrid AgNPs and hybrid CuNPs show correlations between the release of agents and biological activities. Certain nanostructured polymeric coats could complement these NPs if there exhibit a controlled release of the fungicides [85]. Additionally, ZnO-NPs possess few health concerns as there were no immunological or cytotoxic responses when exposed to in vitro cell studies against health human monocytes lines [89]. Favorable cytotoxicity against bacterial cells and healthy human colon cells has been shown with the use of polymer-AgNPs [88]. The administration of the nanofungicides is typically imagined analogous to the current method, in a liquid spray; however, other methods are being explored. Silica-Ag-carbon-based nanoparticles infused into cotton fabrics demonstrate high antifungal activity [84]. Although many researchers have shown hybrid nanofungicidal use in vitro is successful, these compounds are beginning to be readily used for disinfecting water, food packaging, and food preservation [83,86].

5. Nanofunigicides mode of action against toxigenic fungi The mode of action for nanofungicides is still being assessed today. Because of a large number of different phytochemicals and phytochemical derivatives (see Fig. 7.6, for examples), one can expect a number of different methods for different phytochemicals. Below, some of the most common methods are discussed and comprehensively depicted in Fig. 7.11. Note that some of these can act alone, or supplemental to other actions.

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177 FIGURE 7.10 Hybrid nanofungicides composed of various components (1) metallic alloy for photocatalytic activity, (2) bioactive phytochemical with antifungal activity, (3) antifungal agent loading to increase of antifungal activity, and (4) biopolymer for mycotoxin adsorption.

178 SECTION | II Synthesis, toxicity and management FIGURE 7.11 Modes of antifungal activity against toxigenic fungi facilitated by (1) disturbance of cell wall/membrane integrity, (2) generation of reactive oxygen species, (3) mitochondrial disruption, (4) caspase cascade, and (5) chromatin condensation leading to apoptosis.

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5.1 Disturb cell wall integrity Some phytochemicals inhibit cell wall synthesis or the cell regulatory mechanisms, causing the contents of cells to be dumped and deflate. These cells result in appearing “mulberry-like” with areas that have deep grooves, implying the beginning of the cells collapse. Together, these suggest the phytochemicals induce the formation of pores and inhibit the integrity of the plasma membrane and cell wall [90,91]. Phytochemicals (e.g., eugenol) increase the fluidity and permeability of the membrane of microorganisms. They interfere with ion transport, unbalance osmotic conditions and making the associated proteins embedded in the membrane inefficient [90]. Other phytochemicals (e.g., isoquercitrin) induce the release of dextran from membrane lipids and eventually lead to cell death [92,93].

5.2 ROS accumulation A majority of phytochemicals induce the
excessive production of ROS [94e97] including the superoxide anion O2 $ , hydrogen peroxide (H2O2), and radical hydroxyl (OH,). Large quantities of these ROS are ubiquitous with apoptosis and are used as early detectors for apoptosis [98e100]. Pathogenic microbes exposed to phytochemicals show significantly larger numbers of ROS versus a control. Under normal mitochondrial respiration, O2 $ is generated then subsequently converted into H2O2 by superoxide dismutase [101]. H2O2 interacts with Feþ2 to give OH, which is extremely reactive (see Fenton and HaberWeiss reactions) [102]. OH, nonselectively interacted with cellular components like fatty acids, amino acids, and DNA. Moreover, when introduced to cells, phytochemicals yield very high levels of the OH, [103,104].

5.3 Mitochondrial disruption Phytochemicals have been shown to interact with mitochondria in ways that are characteristic of apoptosis: increase ROS, decrease mitochondrial membrane potential, and release cytochrome c and lactate dehydrogenase [105]. The release of cytochrome c (and the apoptosis-inducing factor) into the cytosol causes the degradation of mitochondrial membranes, depolarization of mitochondrial membrane, and ultimately mitochondrial fragmentation [106]. Normally, cytochrome c is found within the mitochondrial membrane; however, an increase in ROS presence causes the mitochondria to detach cytochrome c from the inner membrane and extrude it into the cytosol. In the cytosol, cytochrome c activates caspase-9, which is characteristic of the cascade that finalizes the apoptosis process [107,108].

5.4 Caspase cascade Caspases are activated at the early stages of apoptosis. Certain phytochemicals such as amentoflavone and curcumin have been shown to significantly increase

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the presence of caspases and induce their activation [94,98]. Whether the activation of the cascade is a runoff of cytochrome c leakage or direct action by the phytochemicals themselves, the end result is the cascade that leads to cell death.

5.5 Phosphatidylserine externalization, DNA fragmentation, and chromatin condensation When apoptosis is induced, phosphatidylserine (PS) is transported from inner leaflet of the cell’s membrane to the outer leaflet, so the presence of PS on the membrane’s external leaflet implies apoptosis has begun [90]. When treated with certain phytochemicals (e.g., berberine), PS is translocated from the inner to outer leaflet without disturbing the membrane permeability [95]. At the latter stages of apoptosis, DNA begins to fragment, and the cell’s nucleus condenses. When exposed to a-tomatine (highly fungicidal) DNA became highly fragmented and inhibited the growth of fungi whose membranes are low in sterols [98]. Additionally, cells stained with a dye that is used to examine cells in late apoptosis were treated with a-tomatine. After treatment, the high fluorescence of the dye indicated nuclear condensation and, ultimately, cell death [98,104,106].

5.6 Other activity: biofilm growth inhibition A less discussed mode of fungicidal activity that extends beyond an individual cell is the action against biofilms. Phytochemicals from aromatic plants such as oregano and thyme demonstrated high fungicidal activity against fungal biofilms, regardless of the stage of the biofilm’s life (adhesion, growth, maturation) [106e108]. This suggests that even in the late stages of a fungal invasion, phytochemicals still possess the ability to inhibit malignant actions (Fig. 7.11).

6. Conclusion The role of Green Nanotechnology through the utilization of phytochemicals has immensely improved a number of sectors such as applications in medicine, energy, and most recently within the agricultural sector. The manifestation of fungal growth and mycotoxin production is a challenge due to the huge agricultural losses associated with it, and the use of conventional fungicides has proven to be ineffective. It is imperative that the fungicides used have miniscule effects on the individual using them and the surrounding environment. Toxic fungi contaminate human food and animal feed and have tremendous economic impacts on the world. Many acute and chronic ailments can occur due to ingesting contaminated foods and food products. About one-quarter of all crops worldwide are lost due to fungal contamination, annually. This leads to

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detrimental effects on poor nations relying on agriculture both for domestic food sources and exports as their main source of revenue, in addition to nations importing all of these contaminated crops. There exist a plethora of phytochemicals within plants that offer enormous potential to not only fight the malicious fungi but mediate the green synthesis of nanomaterials. The green nanomaterials reviewed mitigate the current issue of lack of bioactivity that phytochemicals possess when considered individually. These fungicidal nanocompounds can include metal-based, polymer-based, or hybrid materials in conjunction with phytoactive compounds. Green Nanotechnological compounds derived from nature can introduce a new era of fungicides that eliminate major agricultural challenges we face today, as well as preventing future famine, food contamination, and ecological degradation.

References [1]

P. Kaur, R. Thakur, A. Choudhary, An in vitro study of the antifungal activity of silver/ chitosan nanoformulations against important seed borne pathogens, Int. J. Sci. Technol. Res. 6 (2012) 83e86. [2] A. Alshannaq, J.H. Yu, Occurrence, toxicity, and analysis of major mycotoxins in food, Int. J. Environ. Res. Public Health. 14 (6) (2017) 632. [3] H. Shanakhat, A. Sorrentino, A. Raiola, A. Romano, P. Masi, S. Cavella, Current methods for mycotoxins analysis and innovative strategies for their reduction in cereals: an overview, J. Sci. Food Agric. 98 (11) (2018) 4003e4013. [4] M.J. Kasprowicz, M. Kozioł, A. Gorczyca, The effect of silver nanoparticles on phytopathogenic spores of Fusarium culmorum, Can. J. Microbiol. 56 (3) (2010) 247e253. [5] M. Saminathan, J. Selamat, A. Abbasi Pirouz, N. Abdullah, I. Zulkifli, Effects of nano-composite adsorbents on the growth performance, serum biochemistry, and organ weights of broilers fed with aflatoxin-contaminated feed, Toxins (Basel) 10 (9) (2018) 345. [6] J. Park, H. Chang, D. Kim, S. Chung, C. Lee, Long-term occurrence of deoxynivalenol in feed and feed raw materials with a special focus on South Korea, Toxins (Basel) 10 (3) (2018) 1e14. [7] R. Ma, L. Zhang, M. Liu, Y.T. Su, N.Y. Zhang, J.F. Dai, Y. Wang, S.A. Rajput, D.S. Qi, N.A. Karrow, L.H. Sun, Individual and combined occurrence of mycotoxins in feed ingredients and complete feeds in China, Toxins (Basel) 10 (3) (2018) 1e13. [8] F. Vanara, V. Scarpino, M. Blandino, Fumonisin distribution in maize dry-milling products and by-products: impact of two industrial degermination systems, Toxins (Basel) 10 (9) (2018) 357. [9] N. Nleya, M.C. Adetunji, M. Mwanza, Current status of mycotoxin contamination of food commodities in Zimbabwe, Toxins (Basel) 10 (5) (2018) 1e12. [10] Q.S. Yuan, P. Yang, A.B. Wu, D.Y. Zuo, W.J. He, M.W. Guo, T. Huang, H.P. Li, Y.C. Liao, Variation in the microbiome, trichothecenes, and aflatoxins in stored wheat grains in wuhan, China, Toxins (Basel) 10 (5) (2018) 1e14. [11] S. De Saeger, K. Audenaert, S. Croubels, Report from the 5th international symposium on mycotoxins and toxigenic moulds: challenges and perspectives (MYTOX) held in Ghent, Belgium, May 2016, Toxins (Basel) 8 (5) (2018) 146. [12] N. Aslam, M.Y. Tipu, M. Ishaq, A. Cowling, D. McGill, H.M. Warriach, P. Wynn, Higher levels of aflatoxin M1 contamination and poorer composition of milk supplied by informal milk marketing chains in Pakistan, Toxins (Basel) 8 (12) (2016) 347.

182 SECTION | II Synthesis, toxicity and management [13] S. Ashiq, M. Hussain, B. Ahmad, Natural occurrence of mycotoxins in medicinal plants: a review, Fungal Genet. Biol. 66 (2014) 1e10. [14] I.K. Lappa, D. Kizis, E.Z. Panagou, Monitoring the temporal expression of genes involved in ochratoxin a production of Aspergillus carbonarius under the influence of temperature and water activity, Toxins (Basel) 9 (10) (2017) 296. [15] B. Paola, C.L. Marco, OTA-grapes, A mechanistic model to predict ochratoxin a risk in grapes, a step beyond the systems approach, Toxins (Basel) 7 (8) (2015) 3012e3029. [16] D. Lerda, Ochratoxin a (OTA) and public health, in: Ochratoxins : Biosynthesis, Detection and Toxicity, NOVA, 2015, pp. 38e56. [17] R. Santiago, A. Cao, A. Butro´n, Genetic factors involved in fumonisin accumulation in maize kernels and their implications in maize agronomic management and breeding, Toxins (Basel) 7 (8) (2015) 3267e3296. [18] M. Bryła, A. Waskiewicz, K. Szymczyk, R. Jedrzejczak, Effects of pH and temperature on the stability of fumonisins in Maize products, Toxins (Basel) 9 (3) (2017) 1e16. [19] A. Szabo´, J. Szabo-Fodor, H. Febel, M. Mezes, K. Balogh, G. Bazar, D. Kocso, O. Ali, M. Kovacs, Individual and combined effects of fumonisin B1, deoxynivalenol and zearalenone on the hepatic and renal membrane lipid integrity of rats, Toxins (Basel) 10 (2018) 1e17. [20] C. Pietsch, S. Kersten, H. Valenta, S. Danicke, C. Schulz, P. Burkhardt-Holm, R. Junge, Effects of dietary exposure to zearalenone (ZEN) on carp (Cyprinus carpio L.), Toxins (Basel) 7 (9) (2015) 3465e3480. [21] J. Zhang, H. Zhang, S. Liu, W. Wu, H. Zhang, Comparison of anorectic potencies of type a trichothecenes T-2 toxin, HT-2 toxin, diacetoxyscirpenol, and neosolaniol, Toxins (Basel) 10 (5) (2018) 1e14. [22] U. Wachowska, D. Packa, M. Wiwart, Microbial inhibition of Fusarium pathogens and biological modification of trichothecenes in cereal grains, Toxins (Basel) 9 (12) (2017) 1e14. [23] G.D. Savi, A.J. Bortoluzzi, V.M. Scussel, Antifungal properties of Zinc-compounds against toxigenic fungi and mycotoxin, Int. J. Food Sci. Technol. 48 (9) (2013) 1834e1840. [24] S. Gaige´, M. Djelloul, C. Tardivel, C. Airault, B. Felix, A. Jean, B. Lebrun, J.D. Troadec, M. Dallaporta, Modification of energy balance induced by the food contaminant T-2 toxin: a multimodal gut-to-brain connection, Brain. Behav. Immun. 37 (2014) 54e72. [25] M.C. Ferreras, J. Benavides, C. Garcia-Pariente, L. Delgado, M. Fuertes, M. Munoz, J.F. Garcia-Marin, V. Perez, Acute and chronic disease associated with naturally occurring T-2 mycotoxicosis in sheep, J. Comp. Pathol. 148 (2e3) (2013) 236e242. [26] L. Bruno, C. Tardivel, B. Felix, A. Abysique, J.D. Troadec, S. Gaige, M. Dallaporta, Dysregulation of energy balance by trichothecene mycotoxins: mechanisms and prospects, Neurotoxicology 49 (2015) 15e27. [27] T.S. Anjorin, E.A. Salako, H.A. Makun, Control of toxigenic fungi and mycotoxins with Phytochemicals : potentials and challenges, Mycotoxin Food Saf. Dev. Ctries. (2013) 181e202. [28] M. Razzaghi-Abyaneh, A. Allameh, T. Tiraihi, M. Shams-Ghahfarokhi, M. Ghorbanian, Morphological alterations in toxigenic Aspergillus parasiticus exposed to neem (Azadirachta indica) leaf and seed aqueous extracts, Mycopathologia 159 (4) (2005) 565e570. [29] K.R.N. Reddy, C.R. Raghavender, B.N. Reddy, B. Salleh, Biological control of Aspergillus flavus growth and subsequent aflatoxin B 1 production in sorghum grains, Afr. J. Biotechnol. 9 (27) (2010) 4247e4250.

Green nanotechnology Chapter | 7

183

[30] K.M. Soliman, R.I. Badeaa, Effect of oil extracted from some medicinal plants on different mycotoxigenic fungi, Food Chem. Toxicol. 40 (11) (2002) 1669e1675. [31] M. Shehata, A. Badr, A. Abdel-Razek, M. Hassanein, H. Amra, Oil-bioactive films as an antifungal application to save post-harvest food crops, Annu. Res. Rev. Biol. 16 (4) (2017) 1e16. [32] L. Wu, P. Liao, L. He, Z. Feng, W. Ren, J. Yin, J. Duan, T. Li, Y. Yin, Dietary L-arginine supplementation protects weanling pigs from deoxynivalenol-induced toxicity, Toxins (Basel) 7 (4) (2015) 1341e1354. [33] A. Kumar, R. Shukla, P. Singh, N.K. Dubey, Chemical composition, antifungal and antiaflatoxigenic activities of Ocimum sanctum L. essential oil and its safety assessment as plant based antimicrobial, Food Chem. Toxicol. 48 (2) (2010) 539e543. [34] V.K. Reddy, M. Srinivas, A.R. Reddy, G. Srujana, M. Surekha, S.M. Reddy, Plant extracts in the management of aflatoxin production by Aspergillus flavus, Int. J. Pharma Bio Sci. 2 (2) (2011) 492e498. [35] J.S. Dambolena, M.P. Zunino, A.G. Lopez, H.R. Rubinstein, J.A. Zygadlo, J.W. Mwangi, G.N. Thoithi, I.O. Kibwage, J.M. Mwalukumbi, S.T. Kariuki, Essential oils composition of Ocimum basilicum L. and Ocimum gratissimum L. from Kenya and their inhibitory effects on growth and fumonisin production by Fusarium verticillioides, Innov. Food Sci. Emerg. Technol. 11 (2) (2010) 410e414. [36] L. Petruzzi, A. Baiano, A. De Gianni, M. Sinigaglia, M. Corbo, A. Bevilacqua, Differential adsorption of ochratoxin a and anthocyanins by inactivated yeasts and yeast cell walls during simulation of wine aging, Toxins (Basel) 7 (10) (2015) 4350e4365. [37] Y. Li, Y.C. Zhou, M.H. Yang, Z. Ou-Yang, Natural occurrence of citrinin in widely consumed traditional Chinese food red yeast rice, medicinal plants and their related products, Food Chem. 132 (2) (2012) 1040e1045. [38] S.T. Anjorin, H. a Makun, T. Adesina, I. Kudu, Effects of Fusarium verticilloides, its metabolites and neem leaf extract on germination and vigour indices of maize (Zea mays L.), Afr. J. Biotechnol. 7 (14) (2008) 2402e2406. [39] M. Shenashen, A. Derbalah, A. Hamza, A. Mohamed, S. El Safty, Antifungal activity of fabricated mesoporous alumina nanoparticles against root rot disease of tomato caused by Fusarium oxysporium, Pest Manag. Sci. 73 (6) (2017) 1121e1126. [40] S.T. Khalili, A. Mohsenifar, M. Beyki, S. Zhaveh, T. Rahmani-Cherati, A. Abdollahi, M. Bayat, M. Tabatabaei, Encapsulation of Thyme essential oils in chitosan-benzoic acid nanogel with enhanced antimicrobial activity against Aspergillus flavus, LWT - Food Sci. Technol. 60 (2015) 502e508. [41] S.S. Boxi, K. Mukherjee, S. Paria, Ag doped hollow TiO 2 nanoparticles as an effective green fungicide against Fusarium solani and Venturia inaequalis phytopathogens, Nanotechnology 27 (8) (2016), 085103. [42] J. Zhao, L. Wang, D. Xu, Z. Lu, Involvement of ROS in nanosilver-caused suppression of aflatoxin production from Aspergillus flavus, RSC Adv. 7 (37) (2017) 23021e23026. [43] A.M. Rashed, A.A.R. Mohamed, M.M. Abobakr, Wheat protection from root rot caused by Fusarium culmorum using silver nanoparticles, J. Chem. Soc. Pak. 38 (5) (2016) 898e903. [44] M. Razzaghi-Abyaneh, M. Shams-Ghahfarokhi, A. Eslamifar, O.J. Schmidt, R. Gharebaghi, M. Karimian, A. Naseri, M. Sheikhi, Inhibitory effects of AkacidÒplus on growth and aflatoxin production by Aspergillus parasiticus, Mycopathologia 161 (4) (2006) 245e249.

184 SECTION | II Synthesis, toxicity and management [45] S. Majumder, N.A. Shakil, J. Kumar, T. Banerjee, P. Sinha, B.B. Singh, P. Garf, Ecofriendly PEG-based controlled release nano-formulations of Mancozeb: synthesis and bioefficacy evaluation against phytopathogenic fungi Alternaria solani and Sclerotium rolfsii, J. Environ. Sci. Heal. Part B 51 (12) (2016) 873e880. [46] H. Li, Q. Shen, W. Zhou, H. Mo, D. Pan, L. Hu, Nanocapsular dispersion of cinnamaldehyde for enhanced inhibitory activity against aflatoxin production by Aspergillus flavus, Molecules 20 (4) (2015) 6022e6032. [47] A. Kheiri, S.A. Moosawi Jorf, A. Malihipour, H. Saremi, M. Nikkhah, Synthesis and characterization of chitosan nanoparticles and their effect on Fusarium head blight and oxidative activity in wheat, Int. J. Biol. Macromol. 102 (2017) 526e538. [48] K.P. Yu, Y.T. Huang, S.C. Yang, The antifungal efficacy of nano-metals supported TiO2and ozone on the resistant Aspergillus Niger spore, J. Hazard. Mater. 261 (2013) 155e162. [49] M. Beyki, S. Zhaveh, S.T. Khalili, T. Rahmani-Cherati, A. Abollahi, M. Bayat, M. Tabatabaei, A. Mohsenifar, Encapsulation of Mentha piperita essential oils in chitosancinnamic acid nanogel with enhanced antimicrobial activity against Aspergillus flavus, Ind. Crops Prod. 54 (2014) 310e319. [50] S. Zhaveh, A. Mohsenifar, M. Beiki, S.T. Khalili, A. Abdollahi, T. Rahmani-Cherati, M. Tabatabaei, Encapsulation of Cuminum cyminum essential oils in chitosan-caffeic acid nanogel with enhanced antimicrobial activity against Aspergillus flavus, Ind. Crops Prod. 69 (2015) 251e256. [51] R.J.B. Pinto, A. Almeida, S.C. Fernandes, C.S. Freire, A.J. Silvestre, C.P. Neto, T. Trindade, Antifungal activity of transparent nanocomposite thin films of pullulan and silver against Aspergillus Niger, Colloids Surfaces B Biointerfaces 103 (2013) 143e148. [52] E. Babaei, A. Dehnad, N. Hajizadeh, H. Valizadeh, S.F.S. Reihani, A study on inhibitory effects of titanium dioxide nanoparticles and its photocatalytic type on Staphylococcus aureus, Escherichia coli and Aspergillus flavus, Appl. Food Biotechnol. 3 (2) (2016) 115e123. [53] S.I.I. Abdel-Hafez, N.A. Nafady, I.R. Abdel-Rahim, A.M. Shaltout, J.A. Daros, M.A. Mohamed, Assessment of protein silver nanoparticles toxicity against pathogenic Alternaria solani, 3 Biotech. 6 (2) (2016) 199. [54] C. Mitra, P.M. Gummadidala, K. Afshinnia, R.C. Merrifield, M. Baalousha, J.R. Lead, A. Chanda, Citrate-coated silver nanoparticles growth-independently inhibit aflatoxin synthesis in Aspergillus parasiticus, Environ. Sci. Technol. 51 (14) (2017) 8085e8093. [55] A. Sierra-Fernandez, S.C. De la Rosa-Garcia, L.S. Gomez-Villalba, S. Gomez-Cornelio, M.E. Rabanai, R. Fort, P. Quintana, Synthesis, photocatalytic, and antifungal properties of MgO, ZnO and Zn/Mg oxide nanoparticles for the protection of calcareous stone heritage, ACS Appl. Mater. Interfaces 9 (29) (2017) 24873e24886. [56] D. Herna´ndez-Mele´ndez, E. Salas-Te´llez, A. Zavala-Franco, G. Te´llez, A. Me´ndezAlbores, A. Va´zquez-Dura´n, Inhibitory effect of flower-shaped zinc oxide nanostructures on the growth and aflatoxin production of a highly toxigenic strain of Aspergillus flavus link, Materials (Basel) 11 (8) (2018) 1265. [57] A.A. Hassan, M.E. Howayda, H.H. Mahmoud, Effect of zinc oxide nanoparticles on the growth of mycotoxigenic mould, Stud. Chem. Process Technol. 1 (4) (2013) 66e74. [58] G. Sharma, D.T. Valenta, Y. Altman, S. Harvey, H. Xia, S. Mitragotri, J.W. Smith, Polymer particle shape independently influences binding and internalization by macrophages, J. Control. Release 147 (3) (2010) 408e412. [59] S. Barua, J. Yoo, P. Kolhar, A. Wakankar, Y.R. Gokarn, S. Mitragotri, Particle shape enhances specifity of antibody-displaying nanoparticles, Pnas 110 (9) (2013) 3270e3275.

Green nanotechnology Chapter | 7

185

[60] K. Pietrzak, M. Twaruzek, A. Czyzowska, R. Kosicki, B. Gutarowska, Influence of silver nanoparticles on metabolism and toxicity of moulds, Acta Biochim. Pol. 62 (4) (2015) 851e857. [61] H.H. Lara, D.G. Romero-Urbina, C. Pierce, J.L. Lopez-Ribot, M.J. Arellano-Jime´nez, M. Jose-Yacaman, Effect of silver nanoparticles on Candida albicans biofilms: an ultrastructural study, J. Nanobiotechnology 13 (2015) 1e12. [62] D. Gupta, P. Chauhan, Fungicidal activity of silver nanoparticles against Alternaria brassicicola, AIP Conf. Proc. 1724 (2016) 020031e020034. [63] M.M. Deabes, W.K.B. Khalil, A.G. Attallah, T.A. El-Desouky, K.M. Naguib, Impact of silver nanoparticles on gene expression in Aspergillus flavus producer aflatoxin B1. Open access maced, J. Med. Sci. 6 (4) (2018) 600e605. [64] A.M. Abdel-Hadi, M.F. Awad, N.F. Abo-Dahab, M.F. Elkady, Extracellular synthesis of silver nanoparticles by aspergillus terreus: biosynthesis, characterization and biological activity, Biosci. Biotechnol. Res. Asia 11 (3) (2014) 1179e1186. [65] K.K.A. Santos, E.F.F. Matias, S.R. Tintino, C.E.S. Souza, M.F.B.M. Braga, G.M.M. Guedes, J.G.M. Costa, I.R.A. Menezes, H.D.M. Coutinho, Enhancement of the antifungal activity of antimicrobial drugs by Eugenia uniflora L, J. Med. Food. 16 (7) (2013) 669e671. [66] G.D. Savi, W.A. Cardoso, B.G. Furtado, T. Bortolotto, L.O.V. Da Agostin, J. Nones, E.T. Zanoni, O.R.K. Montedo, E. Angioletto, New ion-exchanged zeolite derivatives: antifungal and antimycotoxin properties against Aspergillus flavus and aflatoxin B1, Mater. Res. Express. 4 (8) (2017), 085401. [67] A.A. Pirouz, J. Selamat, S.Z. Iqbal, H. Mirhosseini, R.A. Karjiban, F.A. Bakar, The use of innovative and efficient nanocomposite (magnetic graphene oxide) for the reduction on of Fusarium mycotoxins in palm kernel cake, Sci. Rep. 7 (2017) 1e9. [68] K. Xing, Y. Liu, X. Shen, X. Zhu, X. Li, X. Miao, Z. Feng, X. Peng, S. Qing, Effect of O-chitosan nanoparticles on the development and membrane permeability of Verticillium dahliae, Carbohydr. Polym. 165 (2017) 334e343. [69] M. Sathiyabama, R. Parthasarathy, Biological preparation of chitosan nanoparticles and its in vitro antifungal efficacy against some phytopathogenic fungi, Carbohydr. Polym. 151 (2016) 321e325. [70] G.M. Soliman, Nanoparticles as safe and effective delivery systems of antifungal agents: achievements and challenges, Int. J. Pharm. 523 (2017) 15e32. [71] P. Horky, S. Skalickova, D. Baholet, J. Skladanka, Nanoparticles as a solution for eliminating the risk of mycotoxins, Nanomaterials 8 (9) (2018) 727. [72] A. Fernandes Costa, D. Evangelista Araujo, M. Santos Cebral, I. Teles Brito, L. Borges de Menezes Leite, M. Pereira, A. Correa Amaral, Development, characterization, and in vitroein vivo evaluation of polymeric nanoparticles containing miconazole and farnesol for treatment of vulvovaginal candidiasis, Med. Mycol. (2018) 1e11. [73] C. Bouaoud, S. Xu, E. Mendes, J.G.J.L. Lebouille, H.E.A. De Braal, G.M.H. Meesters, Development of biodegradable polymeric nanoparticles for encapsulation, delivery, and improved antifungal performance of natamycin, J. Appl. Polym. Sci. 133 (31) (2016) 1e11. [74] M.G. De Morais, B.D.S. Vaz, E.G. De Morais, J.A.V. Costa, Biological effects of spirulina (arthrospira) biopolymers and biomass in the development of nanostructured scaffolds, Biomed. Res. Int. (2014) 1e9. [75] Y.H. Shim, Y.C. Kim, H.J. Lee, F. Bougard, P. Dubois, K.C. Choi, C.W. Chung, D.H. Kang, Y.I. Jeong, Amphotericin b aggregation inhibition with novel nanoparticles prepared with

186 SECTION | II Synthesis, toxicity and management

[76]

[77]

[78]

[79]

[80]

[81]

[82]

[83] [84]

[85]

[86] [87] [88]

[89]

[90]

poly(ε-caprolactone)/poly(N,N-dimethylamino-2-ethyl methacrylate) diblock copolymer, J. Microbiol. Biotechnol. 21 (2011) 28e36. B. Gajra, C. Dalwadi, R. Patel, Formulation and optimization of itraconazole polymeric lipid hybrid nanoparticles (Lipomer) using box behnken design, DARU J. Pharm. Sci. 23 (2015) 1e15. K.V. Jardim, A.F. Palomec-Garfias, B.Y.G. Andrade, J.A. Chaker, S.N. Bao, C. MarquezBeltran, S.E. Moya, A.L. Parize, M.H. Sousa, Novel magneto-responsive nanoplatforms based on MnFe2O4nanoparticles layer-by-layer functionalized with chitosan and sodium alginate for magnetic controlled release of curcumin, Mater. Sci. Eng. C. 92 (2018) 184e195. D.C. Pham, T.H. Nguyen, U.T.P. Ngoc, N.T.T. Le, T.V. Tran, D.H. Nguyen, Preparation, characterization and antifungal properties of chitosan-silver nanoparticles synergize fungicide against Pyricularia oryzae, J. Nanosci. Nanotechnol. 18 (8) (2018) 5299e5305. B. Armenda´riz-Barraga´n, N. Zafar, W. Badri, S.A. Galindo-Rodriguez, D. Kabbaj, H. Fessi, A. Elaissari, Plant extracts: from encapsulation to application, Expert Opin. Drug Deliv. 13 (8) (2016) 1165e1175. S. Ilk, N. Saglam, M. Ozgen, Kaempferol loaded lecithin/chitosan nanoparticles: preparation, characterization, and their potential applications as a sustainable antifungal agent, Artifi. Cells Nanomed. Biotechnol. 45 (2017) 907e916. M.M.G. Fouda, M.R. El-Aassar, G.F. El Fawal, E.E. Hafez, S.H.D. Masry, A. AbdelMegeed, K-Carrageenan/poly vinyl pyrollidone/polyethylene glycol/silver nanoparticles film for biomedical application, Int. J. Biol. Macromol. 74 (2015) 179e184. L.-S. Wang, C.Y. Wang, C.H. Yang, C.L. Hsieh, S.Y. Chen, C.Y. Shen, J.J. Wang, K.S. Huang, Synthesis and anti-fungal effect of silver nanoparticles-chitosan composite particles, Int. J. Nanomedicine 10 (2015) 2685e2696. N. Bumbudsanpharoke, S. Ko, Nano-food packaging: an overview of market, migration research, and safety regulations, J. Food Sci. 80 (5) (2015) R910eR923. I. Katerine, A.R. Arreche, E.J. Sambeth, N. Bellotti, J.R. Vega-Baudrit, C. RedondoGomez, P.G. Vazquez, Antifungal activity of cotton fabrics finished modified silicasilver- carbon-based hybrid nanoparticles, Text. Res. J. (2018), 0040517518755792. N. Cioffi, L. Torsi, N. Ditaranto, G. Tantillo, L. Ghibelli, L. Sabbatini, T. Bleve-Zacheo, M. D’Alessio, P.G. Zambonin, E. Traversa, Copper nanoparticle/polymer composites with antifungal and bacteriostatic properties, Chem. Mater. 17 (21) (2005) 5255e5262. A.M. Carmona-Ribeiro, L.D. de Melo Carrasco, Cationic antimicrobial polymers and their assemblies, Int. J. Mol. Sci. 14 (5) (2013) 9906e9946. T. Singh, S. Shukla, P. Kumar, V. Wahla, V.K. Bajpai, Application of nanotechnology in food science: perception and overview, Front. Microbiol. 8 (2017) 1e7. S.K. Verma, E. Jha, K.J. Kiran, S. Bhat, M. Suar, P.S. Mohanty, Synthesis and characterization of novel polymer-hybrid silver nanoparticles and its biomedical study, Mater. Today Proc. 3 (6) (2016) 1949e1957. A. Auyeung, M.A. Casillas-Santana, G.A. Martinez-Castanon, Y.N. Slavin, W. Zhao, J. Asnis, U.O. Hafeli, H. Bach, Effective control of molds using a combination of nanoparticles, PLoS One 12 (2017) e0169940. F. de Oliveira Pereira, J.M. Mendes, E. de Oliveira Lima, Investigation on mechanism of antifungal activity of eugenol against Trichophyton rubrum, Med. Mycol. 51 (5) (2013) 507e513.

Green nanotechnology Chapter | 7

187

[91] M.A.M. Sitheeque, G.J. Panagoda, J. Yau, A.M.T. Amarakoon, U.R.N. Udagama, L.P. Samaranayake, Antifungal activity of black tea polyphenols (catechins and theaflavins) against Candida species, Chemotherapy 55 (3) (2009) 189e196. [92] W. Lee, D.G. Lee, An antifungal mechanism of curcumin lies in membrane-targeted action within Candida albicans, IUBMB Life 66 (11) (2014) 780e785. [93] J. Yun, H. Lee, H.J. Ko, E.R. Woo, D.G. Lee, Fungicidal effect of isoquercitrin via inducing membrane disturbance, Biochim. Biophys. Acta - Biomembr. 1848 (2) (2015) 695e701. [94] J. Lee, D.G. Lee, Novel antifungal mechanism of resveratrol: apoptosis inducer in Candida albicans, Curr. Microbiol. 70 (3) (2015) 383e389. [95] S. Dhamgaye, F. Devaux, P. Vandeputte, N.K. Khandelwal, D. Sanglard, G. Mukhopadhyay, R. Prasad, Molecular mechanisms of action of herbal antifungal alkaloid berberine, in Candida albicans, PLoS One 9 (8) (2014) e104554. [96] M. Sharma, R. Manoharlal, N. Puri, R. Prasad, Antifungal curcumin induces reactive oxygen species and triggers an early apoptosis but prevents hyphae development by targeting the global repressor TUP1 in Candida albicans, Biosci. Rep. 30 (6) (2010) 391e404. [97] H. Choi, D.G. Lee, Lycopene induces apoptosis in Candida albicans through reactive oxygen species production and mitochondrial dysfunction, Biochimie 115 (2015) 108e115. [98] K.-U. Fro¨hlich, F. Madeo, Apoptosis in yeast e a monocellular organism exhibits altruistic behaviour, FEBS Lett. 473 (2000) 6e9. [99] P. Ludovico, F. Madeo, M.T. Silva, Yeast programmed cell death: an intricate puzzle, IUBMB Life 57 (3) (2005) 129e135. [100] G.G. Perrone, S.X. Tan, I.W. Dawes, Reactive oxygen species and yeast apoptosis, Biochim. Biophys. Acta - Mol. Cell Res. 1783 (7) (2008) 1354e1368. [101] H.S. Wong, P.A. Dighe, V. Mezera, P.A. Monternier, M.D. Brand, Production of superoxide and hydrogen peroxide from specific mitochondrial sites under different bioenergetic conditions, J. Biol. Chem. 292 (41) (2017) 16804e16809. [102] B. Halliwell, O.I. Aruoma, DNA damage by oxygen-derived species its mechanism and measurement in mammalian systems, FEBS Lett. 281 (1e2) (2018) 9e19. [103] J.D. Zhang, Z. Xu, Y.B. Cao, H.S. Chen, L. Yan, M.M. An, P.H. Gao, Y. Wang, X.M. Jia, Y.Y. Jiang, Antifungal activities and action mechanisms of compounds from Tribulus terrestris L, J. Ethnopharmacol. 103 (2006) 76e84. [104] A. Rao, Y. Zhang, S. Muend, R. Rao, Mechanism of antifungal activity of terpenoid phenols resembles calcium stress and inhibition of the TOR pathway, Antimicrob. Agents Chemother. 54 (12) (2010) 5062e5069. [105] R. Al Wafai, W. El-Rabih, M. Katerhi, R. Safi, M. El Sabban, O. El-Rifai, J. Usta, Chemosensitivity of MCF-7 cells to eugenol: release of cytochrome-c and lactate dehydrogenase, Sci. Rep. 7 (2017) 1e13. [106] T. Eisenberg, S. Bu¨ttner, G. Kroemer, F. Madeo, The mitochondrial pathway in yeast apoptosis, Apoptosis 12 (5) (2007) 1011e1023. [107] D. Carmona-Gutierrez, T. Eisenberg, S. Bu¨ttner, C. Meisinger, G. Kroemer, F. Madeo, Apoptosis in yeast: triggers, pathways, subroutines, Cell Death Differ. 17 (5) (2010) 763e773. [108] C. Pereira, N. Camougrand, S. Manon, M.J. Sousa, M. Coˆrte-Real, ADP/ATP carrier is required for mitochondrial outer membrane permeabilization and cytochrome c release in yeast apoptosis, Mol. Microbiol. 66 (3) (2007) 571e582.

188 SECTION | II Synthesis, toxicity and management

Further reading [1] P. Kaushik, N.A. Shakil, J. Kumar, M.K. Singh, M.K. Singh, S.K. Yadav, Development of controlled release formulations of thiram employing amphiphilic polymers and their bioefficacy evaluation in seed quality enhancement studies, J. Environ. Sci. Heal. Part B 48 (8) (2013) 677e685.