Nanomycotoxicology: Treating Mycotoxins in the Nano Way

By Mahendra Rai

Nanomycotoxicology: Treating Mycotoxins in Nanoway discusses the role of nanotechnology in the detection, toxicity and management of different types of mycotoxins. Sections cover the topic of nanomycotoxicology, the application of nanotechnology for quicker, more cost-effective and precise diagnostic procedures of mycotoxins and toxicogenic fungi, and the application of nanotechnology for the management of mycotoxigenic fungi. New topics, such as the application of nanotechnology in disease management, disease forecasting, and disease resistance, mycotoxin detection, and nanodiagnostic and molecular techniques are also presented.

With chapter contributions from an international group of experts, this book presents an interdisciplinary reference for scientists and researchers working in mycotoxicology, nanotechnology, mycology, plant science, and food safety. In addition, it will be a useful tool for industrial scientists investigating technologies to update their nanotoxicology and nanosafety knowledge.

• Discusses the role of nanotechnology in mycotoxicology
• Explores the application of nanomaterials for detection of mycotoxins
• Covers the role of nanotechnology in the management of mycotoxins and mycotoxigenic fungi

• Language: English
• Publisher: Academic Press
• Release date: Aug 24, 2019
• ISBN: 9780128179994

Book preview

Nanomycotoxicology

Treating Mycotoxins in the Nano Way

Editors

Mahendra Rai

Nanobiotechnology Laboratory, Department of Biotechnology, SGB Amravati University, Amravati, Maharashtra, India

Department of Chemistry, Federal University of Piaui, Teresina, Brazil

Kamel A. Abd-Elsalam

Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza, Egypt

Table of Contents

Cover image

Title page

Copyright

List of contributors

Preface

Chapter 1. An introduction to nanomycotoxicology

1. Introduction

2. Mycotoxicogenic fungi

3. Mycotoxins

4. Mycotoxicoses

5. Nanomycotoxicology

Section I. Detection

Chapter 2. Role of nanotechnology in the detection of mycotoxins: a smart approach

1. Introduction

2. Important mycotoxins: occurrence and toxicity

3. Conventional methods for the detection of mycotoxins

4. Role of nanotechnology in the detection of mycotoxins: a smart approach

5. Conclusion

Chapter 3. Aptamer-based biosensors for mycotoxin detection

1. Introduction

2. Mycotoxins—general characterization

3. Aptamers for mycotoxin binding

4. Aptasensors for mycotoxin determination

5. Conclusion

Chapter 4. Immunochromatographic techniques for mycotoxin analysis

1. Introduction

2. Immunochromatographic test strip

3. Advantages of lateral flow immunoassay

4. Materials in lateral flow immunoassay

5. Lateral flow immunoassay types

6. Application of immunochromatographic test strip

7. Conclusions

Chapter 5. Magnetic nanomaterials for purification, detection, and control of mycotoxins

1. Introduction

2. Mycotoxin extraction and purification

3. Mycotoxin detection

4. Mycotoxin control

5. Conclusion and future trends

Section II. Synthesis, toxicity, and management

Chapter 6. Mycotoxin-induced toxicities and diseases

1. Introduction

2. Toxicity of aflatoxin

3. Toxicity of ochratoxin

4. Toxicity of fumonisin

5. Toxicity of zearalenone

6. Conclusion

Chapter 7. Green nanotechnology: nanoformulations against toxigenic fungi to limit mycotoxin production

1. Introduction

2. Major mycotoxins

3. Medicinal plants

4. Green chemistry principles

5. Nanofunigicides mode of action against toxigenic fungi

6. Conclusion

Chapter 8. Mycotoxins: decontamination and nanocontrol methods

1. Introduction

2. Prevention of contamination

3. Mycotoxin detoxification

4. A new approach for detoxification with nanoparticles

5. Conclusion

Chapter 9. Heat resistant fungi, toxicity and their management by nanotechnologies

1. The ecology of heat resistant fungi

2. Heat resistance of fungi and the affecting factors

3. Food spoilage caused by heat resistant fungi

4. Growth control of the heat resistant fungi in foods

5. Metabolites of heat resistant fungi

6. Enzymes

7. Mycotoxins

8. Less common heat resistant fungi

9. Nanotechnology and food/feed contamination by fruit-related fungi or their metabolites

10. Nano-aptasensing for analysis of mycotoxins common in fruits

11. Conclusion

Chapter 10. Application of nanoparticles in inhibition of mycotoxin-producing fungi

1. Introduction

2. Mycotoxin-producing fungi

3. Types of nanoparticles and their application for inhibition of fungal growth

4. Action mechanism of nanoparticles against mycotoxic fungi

5. Future perspectives

6. Conclusion

Chapter 11. Metal nanoparticles for management of mycotoxigenic fungi and mycotoxicosis diseases of animals and poultry

1. Introduction

2. Synthesis and characterization of metal nanoparticles

3. Mycosis by Mycotoxic fungi in animal and poultry

4. Mycotoxicosis in animal and poultry

5. Efficacy of metal nanoparticles in management of mycotic diseases in animals

6. Amelioration of the toxic effects of mycotoxins in animals by metal nanoparticles

7. Toxicity of metal nanoparticles

8. Conclusion and future perspectives

Chapter 12. The efficacy of mycotoxin-detoxifying and biotransforming agents in animal nutrition

1. Introduction

2. Controlling mycotoxins by nontoxic mold strains

3. Nanoapproaches for mycotoxin risk elimination

4. Conclusion

Chapter 13. Nanomaterials and ozonation: safe strategies for mycotoxin management

1. Introduction

2. Zinc nanoparticles

3. Silver nanoparticles

4. Selenium nanoparticles

5. Copper nanoparticles

6. Nanoemulsion

7. Nanoadsorbents

8. Superoxide agent (ozonation)

9. Ozonation advantages

10. Ozonizers

11. Ozonation for detoxification of aflatoxins

12. Ozonation for detoxification of trichothecenes

13. Ozonation for detoxification of fumonisin

14. Ozonation for detoxification of zearalenone

15. Ozonation for detoxification of ochratoxin A

16. Conclusion and future trends

Chapter 14. Impact of nanoparticles on toxigenic fungi

1. Introduction

2. Impact of essential oils on toxigenic fungi and production of toxins

3. Impact of metal nanoparticles on toxigenic fungi

4. Nonmetal nanoparticles

5. Carbon-based nanoparticles

6. Conclusions

Chapter 15. Nanocomposites: synergistic nanotools for management of mycotoxigenic fungi

1. Introduction

2. Adverse effects of widespread mycotoxins on human and animal health

3. Polymeric matrices

4. Silica-based NCPs and their hybrids

5. Carbon-based NCPs and their hybrids

6. Metal-based NCPs and their hybrids

7. Conclusions

Chapter 16. Nanotechnological methods for aflatoxin control

1. Introduction

2. Nanoparticles and their properties

3. Effect of nanoparticles on aflatoxin reduction

4. Conclusion

Chapter 17. Antifungal and filmogenic properties of micro- and nanostructures of chitosan and its derivatives

1. Introduction

2. Antifungal activity of chitosan and its oligomers

3. Antifungal activity of micro- and nanostructures of chitosan

4. Postharvest quality of plant products treated with of chitosan

5. Chitosan treatment in some fungal species and changes in hyphal morphology

6. Defense mechanisms in plants elicited by chitosan, microchitosan, nanochitosan, and chitooligomers treatment

7. Antimicrobial properties and mode of action of micro- and nanostructures of chitosan

8. Conclusions and future perspectives

Chapter 18. Nanoparticles and gene silencing for suppression of mycotoxins: what we know and what we should know?

1. Introduction

2. How does RNAi work?

3. RNAi mechanism in management of toxicogenic fungi

4. Transport of siRNA between the host plant cells and the mycotoxigenic fungi

5. Vesicle-mediated RNA transport

6. Transporter-mediated RNA uptake

7. RNAi transmission inside plants

8. RNAi-based approaches in control of toxicogenic fungi

9. RNAi in the field

10. Nanotools to improve RNAi efficiency

11. Conclusion and future perspectives

Chapter 19. Nanostructure self-assembly for direct nose-to-brain drug delivery: a novel approach for cryptococcal meningitis

1. Introduction

2. Cryptococcal meningitis

3. Treatment

4. Flucytosine

5. Azoles

6. General approach

7. Nose-to-brain delivery

8. Nose-to-brain stimuli-responsive systems

9. Liposomes

10. Self-emulsifying drug delivery systems

11. Liquid crystals

12. Dendrimers

13. Nose-to-brain dendrimer drug delivery

14. Concluding remarks and perspectives

Chapter 20. Potent application of nitric oxide–releasing nanomaterials against toxigenic fungi and their mycotoxins

1. Introduction

2. Chemistry and biology of NO

3. NO donors

4. S-nitrosothiols

5. Organic nitrates (nitroglycerin (glyceryl trinitrate) and isosorbide mononitrate)

6. Sodium nitroprusside

7. N-diazeniumdiolates

8. NO and nanomaterials

9. Functionalized metallic nanoparticles

10. Porous silica nanoparticles

11. Polymeric nanoparticles

12. Dendrimers

13. Micelles

14. NO and fungi

15. How does NO exert its antifungal activity?

16. Conclusions

Index

Copyright

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List of contributors

Kamel A. Abd-Elsalam

Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza, Egypt

Unit of Excellence in Nano-Molecular Plant Pathology Research, Plant Pathology Research Institute, Giza, Egypt

Hassan Almoammar,     National Centre for Biotechnology, King Abdulaziz City for Science and Technology (KACST), Riyadh, Saudi Arabia

Carolina Alves dos Santos,     Laboratory of Biomaterials and Nanotechnology, University of Sorocaba, Sorocaba, São Paulo, Brazil

Pierce Bloebaum

Department of Radiology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Institute of Green Nanotechnology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Nahla A. Bouqellah,     Taibah University, Al-Madinah Al-Munawarah, Saudi Arabia

Kemal Çelik,     Çanakkale Onsekiz Mart University, Agricultural Faculty, Çanakkale, Turkey

Marco Vinícius Chaud,     Laboratory of Biomaterials and Nanotechnology, University of Sorocaba, Sorocaba, São Paulo, Brazil

Ana Maria de Oliveira,     Department of Chemistry, Biotechnology and Bioprocess Engineering, Federal University of São João del Rei, Ouro Branco, Brazil

Enio Nazaré de Oliveira Junior,     Department of Chemistry, Biotechnology and Bioprocess Engineering, Federal University of São João del Rei, Ouro Branco, Brazil

Ahmed M.A. El-Hamaky,     Department of Mycology and Mycotoxins, Animal Health Research Institute, Agriculture Research Center, Cairo, Egypt

Medhat A. El-Naggar

Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza, Egypt

Research Central Laboratory, SAGO, Riyadh, Saudi Arabia

Gennady Evtugyn,     Kazan Federal University, Kazan, Russia

Juliana Ferreira de Souza,     Laboratory of Biomaterials and Nanotechnology, University of Sorocaba, Sorocaba, São Paulo, Brazil

Mohamed Amine Gacem

Laboratory of Ecosystems Protection in Arid and Semi-Arid Area, University of Kasdi Merbah, Ouargla, Algeria

Helmholtz Centre for Infection Research, Braunschweig, Germany

Hiba Gacem,     Epidemiology Service and Preventive Medicine, Hassani Abdelkader University Hospital Center, Faculty of Medicine, University of Djillali Liabes, Sidi-Bel-Abbes, Algeria

Ahmed Ghannouchi,     CIHEAM IAMB, Mediterranean Agronomic Institute of Bari, Bari, Italy

Mária Globanová,     Slovak Medical University in Bratislava, Bratislava, Slovakia

Indarchand Gupta,     Department of Biotechnology, Government Institute of Science, Aurangabad, Maharashtra, India

Atef A. Hassan,     Department of Mycology and Mycotoxins, Animal Health Research Institute, Agriculture Research Center, Cairo, Egypt

Tibor Hianik,     Comenius University, Bratislava, Slovakia

Mohamed I.M. Ibrahim,     Food Toxicology and Contaminants Department, National Research Centre, Cairo, Egypt

Avinash P. Ingle,     Department of Biotechnology, Engineering School of Lorena, University of Sao Paulo, Lorena, SP, Brazil

Josef Jampílek

Department of Analytical Chemistry, Faculty of Natural Sciences, Comenius University in Bratislava, Slovakia

Division of Biologically Active Complexes and Molecular Magnets, Regional Centre of Advanced Technologies and Materials, Faculty of Science, Palacky University, Olomouc, Czech Republic

Priti Jogee

Nanobiotechnology Laboratory, Department of Biotechnology, Sant Gadge Baba Amravati University, Amravati, Maharashtra, India

Department of Botany, Prof. Ramkrishna More Arts, Commerce and Science College, Pune, Maharashtra, India

Alice Raphael Karikachery

Department of Radiology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Institute of Green Nanotechnology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Kavita K. Katti

Department of Radiology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Institute of Green Nanotechnology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Kattesh V. Katti

Department of Radiology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Department of Physics, University of Missouri, Columbia, MO, United States

Biological Engineering, University of Missouri, Columbia, MO, United States

Medical Pharmacology and Physiology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Institute of Green Nanotechnology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Menka Khoobchandani

Department of Radiology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Institute of Green Nanotechnology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Katarína Kráĺová,     Institute of Chemistry, Faculty of Natural Sciences, Comenius University, Bratislava, Slovakia

Renáta Lehotská,     Slovak Medical University in Bratislava, Bratislava, Slovakia

Mohamed A. Mohamed,     Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza, Egypt

Emmanuel Njukwe,     International Institute of Tropical Agriculture (IITA), Bujumbura, Burundi

Noha H. Oraby,     Department of Mycology and Mycotoxins, Animal Health Research Institute, Agriculture Research Center, Cairo, Egypt

Aminata Ould El Hadj Khelil,     Laboratory of Ecosystems Protection in Arid and Semi-Arid Area, University of Kasdi Merbah, Ouargla, Algeria

Elena Piecková,     Slovak Medical University in Bratislava, Bratislava, Slovakia

Joana C. Pieretti,     Center for Natural and Human Sciences (CCNH), Federal University of ABC (UFABC), Santo André, SP, Brazil

Mahendra Rai

Nanobiotechnology Laboratory, Department of Biotechnology, Sant Gadge Baba Amravati University, Amravati, Maharashtra, India

Department of Chemistry, Federal University of Piaui, Teresina, Brazil

Mohamed M. Ramadan,     Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza, Egypt

Thais Francine Ribeiro Alves,     Laboratory of Biomaterials and Nanotechnology, University of Sorocaba, Sorocaba, São Paulo, Brazil

Alessandra Cândida Rios,     Laboratory of Biomaterials and Nanotechnology, University of Sorocaba, Sorocaba, São Paulo, Brazil

Wallace R. Rolim,     Center for Natural and Human Sciences (CCNH), Federal University of ABC (UFABC), Santo André, SP, Brazil

Rasha M. Sayed-Elahl,     Department of Mycology and Mycotoxins, Animal Health Research Institute, Agriculture Research Center, Cairo, Egypt

Amedea B. Seabra,     Center for Natural and Human Sciences (CCNH), Federal University of ABC (UFABC), Santo André, SP, Brazil

Alia Telli

Laboratory of Ecosystems Protection in Arid and Semi-Arid Area, University of Kasdi Merbah, Ouargla, Algeria

Department of Biology, Faculty of Naturel Life and Earth Sciences, University of Ghardaïa, Ghardaïa, Algeria

Velaphi C. Thipe

Department of Chemistry, University of Missouri, Columbia, MO, United States

Institute of Green Nanotechnology, University of Missouri, One Hospital Drive, Columbia, MO, United States

Cecilia Torqueti de Barros,     Laboratory of Biomaterials and Nanotechnology, University of Sorocaba, Sorocaba, São Paulo, Brazil

Patchimaporn Udomkun,     International Institute of Tropical Agriculture (IITA), Bujumbura, Burundi

Preface

Many fungi play noteworthy role in spoilage of crops, fruits, and vegetables as a pathogen or by contamination of the harvested products. Within the distinct phases of pathogenesis, however, these fungi can produce diverse secondary metabolites which can have several lethal effects in both animals and humans by invasion of food chain sometimes directly from plant-based food elements infected with mycotoxins or by indirect contamination from the growth of toxigenic fungi on food.

The most significant toxicogenic fungi include Aspergillus, Penicillium, Fusarium, and Alternaria. A major part of the worldwide people depends on cereals as an essential food, and therefore, there is a huge risk of mycotoxin contamination. Furthermore, mycotoxin contamination often has a significant economic and social impact, particularly when its incidence in agricultural commodities is beyond the regulation limits set by national and transnational establishments. The modern advances in nanotechnology have revealed a wide range of applications in detection, diagnosis, and control of pathogens because of the new properties of nanomaterials. Several articles and patents that deal with applications of nanotechnology tools of food testing and safety for mycotoxin contaminations are being published annually.

With its well-known international team of contributors, Nanomycotoxicology: Treating Mycotoxins in the Nano Way discusses the role of nanotechnology in the detection, toxicity, and management of different types of mycotoxins. This book is organized into 20 chapters in three parts: it opens with an introduction to the topic of nanomycotoxicology; Section I examines the role of nanotechnology in the detection and analysis of mycotoxins; and Section II describes the application of nanotechnology for quicker, more cost-effective, and precise diagnostic procedures as well as the synthesis, toxicity, and management of mycotoxigenic fungi. The book covers new topics such as application of nanotechnology in disease management, disease forecasting, disease resistance, mycotoxin detection, and nanodiagnostic and molecular techniques.

This book is an interdisciplinary reference for scientists and researchers working in the field of mycotoxicology, nanotechnology, mycology, plant science, and food safety and is a useful tool for industrial scientists investigating technologies to update their nanotoxicology and nanosafety knowledge.

It is an excellent introduction to this complex topic or a useful supplement to courses in the field of nanomycotoxicology. The purpose of this book is to provide basic knowledge and information to postgraduate students and scientists interested in the upstream research on food safety aspects such as the role of nanotechnology in mycotoxicology, application of nanomaterials for detection of mycotoxins and, finally, cover the role of nanotechnology in the management of mycotoxins and mycotoxigenic fungi.

We are not able to end without acknowledging the authors, who have made significant contribution to this book. Elsevier publisher, who also offered an incredibly great level of professionalism, reliability, and tolerance during the entire procedure, is likewise significantly commended. We thank Kattie Washington and Pat Gonzalez, publishing process managers, for offering the prospect for publishing this book. Last, and by no means least, we express our gratitude to the expert reviewers for their particular informative commentary on this book chapters.

Chapter 1

An introduction to nanomycotoxicology

Kamel A. Abd-Elsalam¹,⁴, and Mahendra Rai²,³     ¹Plant Pathology Research Institute, Agricultural Research Center (ARC), Giza, Egypt     ²Nanobiotechnology Laboratory, Department of Biotechnology, Sant Gadge Baba Amravati University, Amravati, Maharashtra, India     ³Department of Chemistry, Federal University of Piaui, Teresina, Brazil     ⁴Unit of Excellence in Nano-Molecular Plant Pathology Research, Plant Pathology Research Institute, Giza, Egypt

Abstract

Mycotoxins are secondary metabolites of fungi, which live as parasites or saprophytes on plant foods or livestock forage. They mainly include the members of Aspergillus, Penicillium, and Fusarium. Globally, the population depends on cereals, which are usually contaminated by fungi that produce mycotoxins. The contamination of food crops and products by mycotoxin can have a tremendous financial and social impact. There are various techniques for mycotoxin analysis. These include mainly ELISA test, the lateral flow test, the screening cards, and immunoaffinity columns. Recently, the application of nanotechnology in biosensors has led to the rapid detection of mycotoxins. This chapter is concerned with mycotoxicogenic fungi, mycotoxins, mycotoxicoses, mycotoxicology, and nanomycotoxicology.

Keywords

Aflatoxins; Mycotoxicoses; Nanomycotoxicology; Toxigenic fungi

1. Introduction

According to Food and Agriculture Organization, mycotoxins are responsible for contamination of approximately 25% of the world's food crops. Globally, 100 million tons of the agricultural products, such as maize, peanuts, coconut, betel nuts, and oilseeds, are at high risk, of which 20 million tons come from the developing nations [1]. Mycotoxins are secondary metabolites of fungi, which can be parasites or saprophytes of crop plants or livestock forage. The most important toxicogenic fungi are Aspergillus, Penicillium, Fusarium, and Alternaria. A major part of the universal population in all over the world depends on cereals as a main food, and therefore, there is a high risk of mycotoxin. Moreover, mycotoxin contamination can have a large financial and social impact, specifically when the incidence of mycotoxin in food commodities is beyond the regulation limits set up by national and transnational establishments. There are various techniques for mycotoxin analysis. These include mainly ELISA test, the lateral flow test, the screening cards, and immunoaffinity columns. Lately, the emergence of nanotechnology in biosensors has enabled scientists for detection of mycotoxins rapidly. The development of sensors by using nanomaterials affords exquisite benefits inclusive of miniaturization of devices. These nanomaterial-based sensors are rapid, sensitive, economically viable, and hence useful for the food enterprises for the detection of mycotoxins and maintenance of food quality [2]. The utilization of nanotechnology in maintenance of plant health particularly for the control of secondary metabolites, fast detection and management of the diseases, enhanced capability for uptake of nutrients, and formulation and use of effective nanofertilizers has proved it to be a powerful technology, and therefore, it has the capability to revolutionize food and agriculture [3]. The emergence of nanotechnology and the development of nanoscale materials have made vital changes in agriculture [4,5]. The progress in nanotechnology is responsible for the invention of many methods beneficial for the detection and sensing of mycotoxin in livestock [6–8]. Furthermore, the researchers are involved in the fabrication of most sensitive systems for the detection and control of mycotoxigenic fungi and their toxins [9]. The usage of nanotechnology for improvement of nanobiosensors will be a novel strategy for the fast detection of mycotoxins. The production of nanobiosensors and their use for the recognition of the mycotoxins in food and feed would be immensely helpful [10,11].

Thus, there is a huge demand to develop a feasible approach to manage toxigenic fungi and their mycotoxins. Nanotechnological claims in mycotoxicology are still in its primary stage. Currently, research has been focused on the development of new nanomaterials to inhibit pathogenic fungi and mycotoxins [12]. Nanotechnology can precisely target specific food safety problems in agriculture, such as its application against toxicogenic fungi, and provide new techniques for detection and management of mycotoxins, for example, using bio- and nanosensors for detection of mycotoxins. A new nanobiotechnology method describes a novel plant gene transfer technique for improvement of resistance in crops against plant pathogens to enhance food security. In addition, quantum dots play an important role for rapid recognition of a specific biological marker with great precision. Nanobiosensor, quantum dots, nanoimaging, and nanopore DNA sequencing have proved their potential to increase specificity, sensitivity, and rapidity in disease detection and management of food quality and safety. Now, nanofungicides and nanocomposites are being used widely in agriculture and environmental applications.

Mycotoxicology is the branch of mycology that focuses on analyzing and finding out the secondary metabolite produced by fungi, known as mycotoxins, whereas nanomycotoxicology deals with the treatment of the mycotoxins in nanoway. Throughout the last years, nanoresearchers published 57,558 articles in agriculture, 161,029 articles in foods, and 351,547 articles in environmental nanotechnology. Today, there are more than 847 nanotechnology patents used for identification, purification, and management of aflatoxins, and 679 different types of nanoparticles patent used to identify, purify, and manage different mycotoxins (Fig. 1.1).

2. Mycotoxicogenic fungi

The incidence of fungi and production of mycotoxins in agricultural crops occur under favorable conditions in the field (preharvest), at harvest, and during handling, transport, and storage. Some Fusarium species are communal plant pathogenic fungi occurring globally, particularly associated with cereal crops. There are more than 100 secondary metabolites produced by Fusarium species, a number of which can critically affect human and animal health [13]. Despite the fact that more than 100 species of molds had been described, more than 500 mycotoxins have been diagnosed due to analytical advances in the analysis of many mycotoxins in crop yields [14–16].

Figure 1.1  Number of patents registered and published articles in nanomycotoxicology based on the type of keywords (1   =   Nano, Aflatoxins; 2   =   Nano, Mycotoxins, 3   =   Nanoparticles, Aflatoxins, 4   =   Nanoparticles, Mycotoxins; 5   =   NMs, Aflatoxins, 6   =   NMs, Mycotoxins; 7   =   Nanotechnology, Aflatoxins; 8   =   Nanotechnology, Mycotoxins). 

Data obtained from google patents https://patents.google.com/ and Nano Nature Database https://nano.nature.com/.

3. Mycotoxins

The mycotoxins (Greek mukos or mykes means fungus and toxikon refers to poison) are secondary metabolites secreted by fungi, which inhabit plants during pre- and postharvest period [17]. Mycotoxins are low molecular weight (MW ∼700  Da) secondary metabolites particularly formed by Aspergillus, Fusarium, and Penicillium. These are notably deleterious to animals and human beings. The fungal toxins can be categorized into the following: (1) mycotoxins formed by mycelia of common molds and (2) mushroom toxins produced inside the fruiting bodies of higher fungi. On the basis of their molecular characters, fungal toxins may be divided into two groups: nonpeptidic toxins and poisonous peptidic toxins. Most of the fungi can secrete more than a single mycotoxin, but a given mycotoxin can also be produced by some species that belong to distinct genera. There are certain mycotoxins which have demonstrated acute toxic effects, particularly when consumed at high concentrations, whereas others have toxic effects only after a long-term exposure to lower doses, i.e., chronic effects. The main mycotoxins include aflatoxin B1 (AFB1), aflatoxin G1 (AFG1), ochratoxin A (OTA), deoxynivalenol (DON), nivalenol (NIV), fumonisin (FUM), zearalenone (ZEA), patulin (PAT), and citrinin (CIT), which are mostly produced by Aspergillus, Fusarium, and Penicillium. It has been assessed that the cost of crop loss due to mycotoxins (aflatoxins, fumonisins, and deoxynivalenol) in United States alone is $932  million per year, which is very high. Moreover, medication costs of $466  million and livestock costs of $6  million are additionally required [18].

4. Mycotoxicoses

Mycotoxicoses are human or animal diseases caused by consumption of fungal contaminated foods, skin contact with mold-infested substrates, and inhalation of toxins secreted by fungal spores. Mycotoxins can have an effect on the animals either alone or additively, it's fundamental to differentiate between mycotoxicosis and mycosis. Mycotoxicosis is commonly used to define the effect of mycotoxin (s), which is often facilitated through many organs, particularly kidney, liver, and lungs, and consequently endocrine and immune system, whereas mycosis refers to general infections caused by fungi in human and animals due to different environmental and physiological conditions.

Mycotoxins and mycotoxicoses are an especially oversize problem for human and animal health because below certain conditions crop and foodstuffs can provide a good medium for growth of the fungi leading to toxin production. Exposure to mycotoxins may also generate toxic syndromes on affected parts referred to as mycotoxicoses. Mycotoxicoses occur commonly in tropical regions due to high humidity and temperature required for fungal growth and secretion of mycotoxin [19].

Mycotoxicoses in humanlike other toxicological syndromes can be labeled as acute or chronic. Acute toxicity has a fast onset and an obvious poisonous response, at the same time as continual toxicity is characterized by low dose exposure over a longtime duration leading to cancer and other commonly reversible effects [20]. The extent of detrimental consequences of mycotoxins on the health of people or animals depend on dose and length of exposure, type of mycotoxin, physiological and dietary status, and possible synergistic outcomes of different chemical compounds to which the human beings are exposed [21]. The study concerning the evolution of mycotoxins and mycotoxicoses is receiving considerable attention with the overall thrust for prevention of mycotoxin production and to save our food/feed products from contaminating and fungi-causing mycotoxicoses.

Of approximately 500 recognized mycotoxins, only a small variety are documented to motive mycotoxicoses in human and animals. The organs such as liver and kidneys are mostly affected as these are the ones in which mycotoxins are metabolized; however, they may also affect different systems of the body. Historically, acute mycotoxicoses have been common even in mild temperature zones, causing epidemics that devastated entire regions, from time to time influencing the direction of human documents. Growth taking place on human beings is referred to as mycoses, whereas human pathogenesis as a result of fungal metabolite (toxin) is referred to as mycotoxicoses [22].

The mycotoxins such as patulin, PR toxin, roquefortine, and mycophenolic are known to possess the high potential of causing mycotoxicoses in animals. Among these, PR toxin is thought to have prompted mycotoxicoses in cattle that had consumed Penicillium roqueforti-contaminated feed [23]. Ergotism (mycotoxicosis caused by long-term exposure of ergot alkaloids) is responsible for the deaths of a massive number of human beings and has no longer been visible in temperate zones for hundreds of years [24].

Mycotoxin can cause toxicity, may develop cancer, affect the liver, kidney, and nervous and immune system. In addition, the mycotoxicity may be teratogenic, estrogenic, and hemorrhagic [25]. Mycotoxicoses signs and symptoms rely on the type of mycotoxin; the amount and period of the exposure; the age, fitness, and intercourse of the exposed individual; and several poorly understood synergistic consequences regarding genetics, dietary status, and interactions with other toxic insults. In acute mycotoxicoses, symptoms appear rapidly, and if exposure continues, the final results may be deadly. Chronic mycotoxicosis is the result of longtime exposure to smaller quantities of mycotoxins. The preliminary stage of contact is often insidious and without apparent preliminary symptoms [26].

Fungal genera causing mycotoxicoses are ideally identified by a polyphasic technique, which will keep away from errors, beginning at genus stage and similarly to species degree using a combination of morphological, physiological, nutritional, and chemical statistics. Generally, the identification is confirmed by using PCR-based molecular strategies which may be taken into consideration under two important complementary strategies: by means of focus on conserved functional genes or regions of taxonomical attention or via focusing at the mycotoxigenic genes [27].

5. Nanomycotoxicology

There is a growing need for multidisciplinary reports across both natural and social sciences, for the evolution of sustainable nanotechnology. The study of mycotoxins is a subdiscipline called mycotoxicology, whereas the animal and human diseases caused by mycotoxins are known as mycotoxicoses. As a matter of fact, nanomycotoxicology is a new and emerging science and presently it is at the embryonic stage. Nanomycotoxicology (nanotechnology  = the creation and exploitation of materials in the size range of 1–100  nm, mycotoxicology  =  study of mycotoxins) is a brand new term that is offered here for the first time. There are no books available on applications of nanotechnology for the detection and management of mycotoxins. The foremost goal of this book is to furnish up-to-date knowledge to researchers working on nanotechnology to address the mycotoxin problem. This book has been divided into three sections: Section I introduces mycotoxicology, mycotoxicoses, mycotoxicogenic fungi, mycotoxins, and nanomycotoxicology; Section II deals with nanotechnology for rapid, cost-effective, diagnostic techniques of mycotoxins and toxicogenic fungi; and Section III describes the use of nanotechnology for the management of mycotoxigenic fungi and mycotoxins.

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Section I

Detection

Outline

Chapter 2. Role of nanotechnology in the detection of mycotoxins: a smart approach

Chapter 3. Aptamer-based biosensors for mycotoxin detection

Chapter 4. Immunochromatographic techniques for mycotoxin analysis

Chapter 5. Magnetic nanomaterials for purification, detection, and control of mycotoxins

Chapter 2

Role of nanotechnology in the detection of mycotoxins

a smart approach

Avinash P. Ingle¹, Indarchand Gupta², Priti Jogee³,⁴, and Mahendra Rai³,⁵     ¹Department of Biotechnology, Engineering School of Lorena, University of Sao Paulo, Lorena, SP, Brazil     ²Department of Biotechnology, Government Institute of Science, Aurangabad, Maharashtra, India     ³Nanobiotechnology Laboratory, Department of Biotechnology, Sant Gadge Baba Amravati University, Amravati, Maharashtra, India     ⁴Department of Botany, Prof. Ramkrishna More Arts, Commerce and Science College, Pune, Maharashtra, India     ⁵Department of Chemistry, Federal University of Piaui, Teresina, Brazil

Abstract

Mycotoxins are toxic secondary metabolites produced by certain filamentous fungi. They can cause various hazardous effects in both animals and humans by entering in food chain either directly from plant-based food components contaminated with mycotoxins or by indirect contamination from the growth of toxigenic fungi on food. Different conventional methods are available for the detection of mycotoxins, but their limitations associated with sensitivity and specificity restrict their uses up to some extent. However, the recent progress in nanotechnology has demonstrated wide range of applications in detection, diagnosis, and management of pathogens due to the novel properties of nanomaterials. One of the applications that has emerged in the utilization of nanotechnology in agriculture is the detection of mycotoxins and management of mycotoxigenic fungi. The high surface-to-volume ratio of nanoparticles provides improved support for the devolvement of nanosensors through the immobilization of desired biomolecules for signal amplification for the detection of mycotoxins. Among the various nanomaterials, metallic and magnetic nanoparticles are most preferably used in the fabrication of nanosensors for mycotoxin detection. This chapter provides an overview of mycotoxins including their various types and conventional methods available for the detection of mycotoxins. However, special emphasis has been made on the various nanotechnological approaches used for the detection of mycotoxins.

Keywords

Conventional methods; Mycotoxicoses; Mycotoxins; Nanomaterials; Nanosensors; Nanotechnology

1. Introduction

The term mycotoxin has been evolved from Greek word (μύκης, i.e., mykes, mukos) fungus and Latin word (toxicum) meaning poison It is a toxic secondary metabolite produced by an organisms of the kingdom fungi (mycota), including mushrooms, molds, and yeasts [1,2]. The term mycotoxin is usually reserved for the toxic chemical products produced by fungi that readily colonize in crops [3]. Mycotoxins are the structurally diverse group of mostly small molecular weight compounds, ubiquitous in a broad range of commodities and feed and are toxic to mammals, poultry, and fish [4]. Mycotoxins can cause acute or chronic toxicity in animals and humans owing to ingestion of mycotoxin-contaminated feed or food, which is commonly referred to as mycotoxicoses.

Mycotoxins are a large and diverse group of naturally occurring chemicals, produced by a variety of fungi, but mycotoxins produced by the members of the genera Aspergillus, Penicillium, and Fusarium are of prime importance due to their direct effect on public health. Various feeds and food crops including cereal grains are mostly contaminated with several fungal species, and the initial infestation, growth, and subsequent mycotoxin production mainly occur during the cultivation and/or storage of the crops [5]. Because of the fungal infestation, cell membrane is dissolved, which protects cell from invading organisms. Moreover, the production of mycotoxins depend on a variety of causes including environmental factors such as temperature and moisture at any stage of plant growth [3]. It is not necessary that infestation of mycotoxin-producing fungi in cereal grains should always be conducive to contamination by mycotoxins because it requires stress due to several factors for production of mycotoxins [6].

It is estimated that over 300 mycotoxins have been identified worldwide. However, according to some reports, the approximate number of mycotoxins may be in the range of 20,000 to 300,000 [7–9]. However, regardless of a number of estimated mycotoxins, some groups such as aflatoxins (AFs), ochratoxins (OTs), trichothecenes [T-2 toxin, HT-2 toxin, deoxynivalenol (DON), nivalenol (NIV), diacetoxyscirpenol (DAS)], zearalenone (ZEN), and fumonisins (FBs) are considered as most important due to their major concerns in public health [4]. The diverse chemical nature of these mycotoxins may selectively play important role in their toxicities and the target organs in animals and humans [1]. These mycotoxins generally showed carcinogenic, teratogenic, hepatotoxic, nephrotoxic, and immunosuppressive properties [10]. Among the above-mentioned mycotoxins, AFs are commonly produced by some species of the genus Aspergillus. However, OTs are the secretory product of Aspergillus spp. or Penicillium spp. depending on the type of agriculture commodity and environmental conditions. In addition, some Fusarium species are particularly responsible for the production of trichothecenes, ZEN, and FBs.

Food and Agriculture Organization of the United Nations reported that about 25% of food production worldwide gets contaminated by mycotoxins. Furthermore, because of the ever-growing population, world is facing the challenges of safe food security [11]. From the last many decades, world hunger continues to increase and the number of undernourished people was raised from 777 million to 815 million in 2016. Moreover, the climate change and hot and humid conditions generally favor fungal growth, which leads to increased mycotoxin contamination with detrimental effects, where one-fourth of children under the age of 5  years suffer from stunting because of the intake of contaminated foods [11,12].

Similarly, it was claimed that mycotoxin-related threats to livestock production have augmented in most regions of the world over the first quarter of 2017, and it was proved from the analysis of more than 14,000 samples including 3715 finished feed and raw commodity samples sourced from 54 countries from January to March 2017 [13]. According to this report, during the first half of 2017, the Northern Hemisphere showed the relatively high risk. However, in the second half of 2017, the risk level increased in the northern part of Europe (on average, from 44% increased to 70% of samples at a level above the risk threshold). Increases were also seen in North America (from 59% in the second half of 2016 to 77% in the second half of 2017), East Asia (88%–92%), Oceania (24%–26%), Middle East (from 47% to 69%), and Africa (from 48% to 73%). Fig. 2.1 shows the global map of mycotoxin occurrence and risk in different regions.

Figure 2.1  Global map of mycotoxin occurrence and risk in different regions. 

Adapted from BIOMIN. World Mycotoxin Survey, 2017, Annual Report No. 14; with copyright permission.

It is a well known fact that mycotoxins are most important contaminants in foods and feeds. Therefore, their early and fast detection is essential to maintain good quality of food and to reduce its negative impact on human and animal health. The contamination of food products by mycotoxins can be avoided by monitoring and having control at different critical steps of the food chain in pre- and postharvest stage. It mainly includes monitoring of raw materials and food supply, monitoring during food processing, monitoring of final products, and also during storage [14]. Advanced technologies are required to assess the safety and quality of the foods to achieve a multisensing system that integrates analytical requirements of sensitivity and selectivity in mycotoxin detection. Considering the widespread use of nanotechnology in various sectors, it is believed that the application of many nanomaterials in the detection of mycotoxins will be pathbreaking strategies.

The main objective of this chapter is to describe the role of nanotechnology in the detection of mycotoxins. In addition, a brief discussion has also been provided on various types of mycotoxins, their health hazards, and the conventional methods available for their detection.

2. Important mycotoxins: occurrence and toxicity

As mentioned above, approximately 300 to 300,000 different types of mycotoxins have been investigated from various fungi [7–9]. However, among these, only a few groups of mycotoxins have been reported to cause diseases to human and animals.

2.1. Aflatoxins

AFs are generally produced by some species of Aspergillus, which are broadly divided into three different groups, i.e., aflatoxin B (AFB1 and AFB2) (Fig. 2.2A and B), aflatoxin G (AFG1 and AFG2) (Fig. 2.2C and D), and aflatoxin M (AFM1 and AFM2) [15] (Fig. 2.2E and F). AFs can be found in a wide variety of foods such as cereals (maize, rice, barley, oats, and sorghum), peanuts, groundnuts, pistachio nuts, almonds, walnuts, and cotton seeds [16,17]. Of these, AFB1 is considered as the most health hazardous due to its carcinogenic nature. The biotransformation of AFB1 and AFB2 results in the production of their hydroxylated metabolites, i.e., AFM1 by hepatic microsomal cytochrome P450 in cows fed on contaminated feed. Therefore, AFM1 and AFM2 are mostly found in milk and milk products due to ingestion of Aspergillus flavus and Aspergillus parasiticus-contaminated feed consumed by livestock [18]. AFB1 and AFM1 are classified as a Group 1 carcinogen by International Agency for Research on Cancer (IARC) because it was reported to cause human primary liver cancer synergistically with hepatitis B virus [19]. The first outbreak of AFs toxicity (aflatoxicosis) affecting humans was reported in India, which resulted in the death of 100 people [20]. In addition, high consumption of AFs by children show evidence of growth impairment and stunting, which makes such children more susceptible to other illnesses. High exposure levels of AFs may lead to acute poisoning and even deaths.

Figure 2.2  Chemical structure of various aflatoxins: (A) aflatoxin B 1 , (B) aflatoxin B 2 , (C) aflatoxin G 1 , (D) aflatoxin G 2 , (E) aflatoxin M 1 , (F) aflatoxin M 2 .

2.2. Ochratoxins

OTs are another group of mycotoxins for the first time discovered in South Africa in 1965, and it is produced by different Aspergillus and Penicillium species [21]. It is mainly classified into ochratoxin A (OTA), ochratoxin B (OTB), ochratoxin C (OTC), and ochratoxin α (OTα) [22] (Fig. 2.3A–D). Among these, OTA is reported to be highly toxic, showed nephrotoxic and nephrocarcinogenic effects [23], and is predominantly found in cereals, coffee, dried fruits, spices, grape juice, and animal feeds [24,25]. Moreover, OTs can be found in various animal-derived products such as meat and milk and can be present in human milk [26]. It was proposed that after ingestion, OTA binds to serum albumin [27], and it was suspected to cause Balkan endemic nephropathy which affected southeastern Europeans [28]. It is a renal disease that can cause kidney and liver failure. According to IARC, OTA is a Group 2B carcinogen (possible human carcinogen), which induces oxidative DNA damage and also causes immunotoxicity, genotoxicity, neurotoxicity, teratogenicity, and embryotoxicity in both human and animals [29].

Figure 2.3  Chemical structure of different ochratoxins: (A) ochratoxin A, (B) ochratoxin B, (C) ochratoxin C, (D) ochratoxin α.

2.3. Fumonisins

FBs were discovered in 1988 and belong to non-fluorescent mycotoxin's group [30]. Structurally these mycotoxins are different from most other mycotoxins and are hydrophilic. Moreover, these can be dissolved completely in organic solvents. Fusarium spp. (Fusarium verticillioides and Fusarium proliferatum) are generally responsible for the production of FBs, and it is ubiquitously found in corn. There are different types of FBs, i.e., fumonisin B1 (FB1), fumonisin B2 (FB2), and fumonisin B3 (FB3) (Fig. 2.4A-C), which are predominantly found in contaminated corn [31]. Among all the FBs, FB1 is the most commonly found, and it comprises about 70%–80% of the total FB family. In addition to corn, FBs can also be found in many other crops and medicinal plants including sorghum, wheat, barley, soybean, asparagus spears, figs, black tea, etc. [16]. FBs are reported to have hepatotoxicity, nephrotoxicity, and also affect immune system of both animals and humans. In 1970, an outbreak of leukoencephalomalacia was reported in equine in South Africa, and pulmonary edema was reported in pigs due to consumption of FB-contaminated feed [32]. FB1 is also classified as a Group 2B carcinogen and has been suspected to esophageal cancer in many cases reported in Egypt, China, South Africa, and the United States of America [33].

Figure 2.4  Chemical structure of different types of fumonisins: (A) fumonisin B 1 , (B) fumonisin B 2 , (C) fumonisin B 3 .

2.4. Trichothecenes

Trichothecenes are another class of mycotoxins, which are classified into two groups (type A and type B trichothecenes). The type A trichothecenes mainly includes T-2, HT-2, and DAS (Fig. 2.5A–C) as mycotoxin components, whereas type B trichothecenes include DON, NIV, 3-acetyldeoxynivalenol, and 15-acetyldeoxynivalenol [34] (Fig. 2.6A–D). Above mentioned trichothecenes are usually produced by a variety of Fusarium species. However, some other fungi such as Acremonium (Cephalosporium), Cylindrocarpon, Dendrodochium, Myrothecium, Trichoderma, Trichothecium, and Stachybotrys were also reported to produce trichothecenes [35,36].

The fungal species responsible for these mycotoxins are mainly colonies in cereal crops such as wheat, barley, oats, rye, maize, and rice [32]. In addition, these mycotoxins are also found in some other crops such as soybeans, potatoes, sunflower seeds, peanuts, bananas, and in some processed foods derived from cereals such as bread, breakfast cereals, noodles, and beer [16]. Trichothecenes showed the toxic effects such as poisoning in both humans and animals. It was also responsible for anorexia, gastroenteritis, emesis, and hematological disorders [37]. Trichothecenes like T-2 and DAS reported to have cytotoxic and immunosuppressive effect in human beings, which results in decrease in resistance to infectious microbes [38,39]. Moreover, T-2 and HT-2 toxins showed higher myelotoxicity (bone marrow damage), inhibition of protein synthesis, and reduction in white blood cells [18,40]. Chickens were found to be more sensitive to trichothecenes than ruminants and pigs [41]. Some other general symptoms of trichothecenes toxicity in animals include weight loss, decreased feed transformation, feed refusal, vomiting, bloody diarrhea, severe dermatitis, hemorrhage, declined egg production, abortion, and death [41–43].

Figure 2.5  Chemical structure of type A trichothecenes: (A) T-2 toxin, (B) HT-2 toxin, (C) Diacetoxyscirpenol.

Figure 2.6  Chemical structure of type B trichothecenes: (A) deoxynivalenol, (B) nivalenol, (C) 3-acetyldeoxynivalenol, (D) 15-acetyldeoxynivalenol.

2.5. Zearalenone

ZEN is another mycotoxin produced by Fusarium species such as Fusarium graminearum and Fusarium semitectum [44] (Fig. 2.7). Structurally, it is very close to naturally occurring estrogens; hence, it is also now regarded as an estrogenic mycotoxin that induces estrogenic effects in human and animals [16]. It is mainly found in crops such as corn, wheat, barley, sorghum, and rye. The high humidity and low-temperature conditions are favorable for the production of ZEN. It is considered as quite stable at regular cooking temperatures; however, elimination of this mycotoxin requires high temperatures [45].

Figure 2.7  Chemical structure of zearalenone.

This mycotoxin is considered as a Group 3 carcinogen by IARC. The health concerns in public due to ZEN are associated with its strong estrogenic activity. It is proposed that generally it binds competitively to some estrogen receptors in various animals leading to changes and lesions in the female reproductive system [46]. In addition, it has the ability to displace the estradiol from its uterine-binding protein, eliciting an estrogenic response [32]. ZEN was discovered as the causative agent of reproductive disorder in pigs known as vulvovaginitis [47]. It also causes reproductive problems in cattle and sheep [48]. It has been used to treat postmenopausal symptoms in women [49]; hence, it was used as oral contraceptives [50]. Other health problems such as infertility, swelling of the uterus and vulva, increased embryo lethal resorptions, and atrophy of ovaries was also reported in various animals such as mice, rats, guinea pigs, and rabbits [51].

Considering the global toxicological effects of abovementioned mycotoxins, various national and international institutions like US Food and Drug Administration (FDA), World Health Organization, Food Agriculture Organization, and the European Food Safety Authority are paying serious attention to mycotoxin contamination in food and feed and released very strict regulatory guidelines for major mycotoxin classes in food and feed [16]. Till date, more than 100 countries have established limits on the presence of these mycotoxins in food and feed [5,15,52]. Table 2.1 shows the list of these important mycotoxins, their sources, and some commonly contaminated food commodities along with the US FDA and EU regulatory limits for mycotoxin levels in food and animal feed.

3. Conventional methods for the detection of mycotoxins

Contamination of food commodities by mycotoxigenic fungi and presence of mycotoxins in food and feed is well studied; however, the problem persists to its detection. Although, there are different traditional methods used for the detection of mycotoxins, the development of new methods is a pressing need. Some techniques used in general for mycotoxin detection are ranged from chromatographic and immunochemical-based techniques to some microarrays performed for the detection of multiple mycotoxins simultaneously. All these techniques need prerequisites for the determination of mycotoxin contamination level in food, feed, and food products: these are sampling, extraction, and cleanup methods [53]. Sampling is very important for determination of mycotoxins. This is because fungal growth is not uniform in the whole sample and may give some misleading results. To overcome such problems, Commission Regulation (EC) No 401/2006 was established for standardization of sampling procedure in the testing of mycotoxins [54]. In the extraction method, mycotoxin is extracted with a suitable solvent and further analysis is carried out. In this procedure, the solvent and the method used for the extraction of mycotoxins from foodstuffs plays a vital role. Zhang et al. [53] have enlisted solvents that are generally used for extraction of mycotoxins. Usually, solid–liquid extraction method is used for extraction of mycotoxins that includes shaking, vertexing, blending, homogenization, and ultrasonic extraction. The choice of method is dependent on the type of sample matrix [55]. The last step is a cleanup method, which is required for concentrating the extracted mycotoxins and to remove any impurity if present. For this purpose, mostly solid phase extraction and immunoaffinity column are in use [56,57].

Table 2.1

The chromatographic methods widely used for the detection of mycotoxins are thin layer chromatography (TLC), liquid chromatography, high performance/high pressure liquid chromatography (HPLC), and gas chromatography often coupled to an ultraviolet detector, fluorescence detector (FLD), or mass spectrometric detector for the quantification of mycotoxin [53]. Wu et al. [58] have reviewed the use of imaging and spectroscopic techniques for the detection of mycotoxigenic fungi present in nuts and some dried fruits. According to the authors, near-infrared spectroscopy (NIRS), mid-infrared spectroscopy, fluorescence spectroscopy/imaging, hyperspectral imaging (HSI), and conventional imaging techniques (color imaging) are mostly used for quantification of mycotoxins.

Another group of scientists has evaluated multiplex polymerase chain reaction (mPCR) assays for the detection of the presence of mycotoxin ZEN in two varieties of rice, i.e., white rice and brown rice, which is commonly secreted by fungi Fusarium spp [59]. The authors have used a set of four primers targeting the ZEA biosynthesis genes, PKS3, PKS13, ZEB1, and ZEB2; in addition, two methods of PCR amplifications were employed that are indirect (DNA isolated from fungi and then amplified) and direct (amplification of target DNA directly from rice samples). And both the mPCR methods had shown good detection ability. These results were then confirmed by comparing with HPLC results, and authors had shown that the mPCR technique is a good alternative for the detection of ZEN mycotoxins. Similarly, another author has given nondestructive methods for the detection of mycotoxins, e.g., NIRS, NIR HSI, and electronic nose (E-nose) [60], whereas for the same purpose, Ge et al. [61] used terahertz spectroscopy. HPLC with an FLD (HPLC-FLD) and photochemical reaction device was developed by Kim et al. [62] for the detection of all four types of aflatoxins (B1, B2, G1,