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Functional and medicinal beverages
 9780128172636, 0128172630, 9780128163979

Table of contents :
Content: 1. An Overview of Functional Beverages 2. The Emerging Trends of Functional and Medicinal Beverage Research and its Health Implication 3. Herbal Extracts --
New Trends in Functional and Medicinal Beverages 4. Bioactive Compounds Incorporated into Functional Beverages 5. Effects and Mechanisms of Antioxidant Rich Functional Beverages on Disease Prevention 6. Anti-Cancer Potential of Functional and Medicinal Beverages 7. Medicinal Properties and Functional Components of Beverages 8. Development of Functional Beverages: The Case of Plant Sterol-Enriched Milk-Based Fruit Beverages 9. Herbal Beverages and Brain Function in Health and Disease 10. Functional Beverages from Cereals 11. Oak as a New Potential Source for Functional Beverages: Their Antioxidant Capacity and Monomer Flavonoid Composition 12. Kombucha as a Functional Beverage 13. Probiotic and Prebiotic Beverages 14. Probiotic Beverages: A Treatment Alternative of Metabolic Syndrome 15. A New Generation of Probiotic Functional Beverages Using Bioactive Compounds from Agro-Industrial Waste.

Citation preview

FUNCTIONAL AND MEDICINAL BEVERAGES

FUNCTIONAL AND MEDICINAL BEVERAGES Volume 11: The Science of Beverages Edited by

ALEXANDRU MIHAI GRUMEZESCU ALINA MARIA HOLBAN

An imprint of Elsevier

Woodhead Publishing is an imprint of Elsevier The Officers’ Mess Business Centre, Royston Road, Duxford, CB22 4QH, United Kingdom 50 Hampshire Street, 5th Floor, Cambridge, MA 02139, United States The Boulevard, Langford Lane, Kidlington, OX5 1GB, United Kingdom © 2019 Elsevier Inc. All rights reserved. No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, i­ncluding photocopying, recording, or any information storage and retrieval system, without permission in writing from the publisher. Details on how to seek permission, further information about the Publisher’s permissions policies and our arrangements with organizations such as the Copyright Clearance Center and the Copyright Licensing Agency, can be found at our website: www. elsevier.com/permissions. This book and the individual contributions contained in it are protected under copyright by the Publisher (other than as may be noted herein). Notices Knowledge and best practice in this field are constantly changing. As new research and experience broaden our understanding, changes in research methods, professional practices, or medical treatment may become necessary. Practitioners and researchers must always rely on their own experience and knowledge in evaluating and using any information, methods, compounds, or experiments described herein. In using such information or methods they should be mindful of their own safety and the safety of others, including parties for whom they have a professional responsibility. To the fullest extent of the law, neither the Publisher nor the authors, contributors, or editors, assume any liability for any injury and/or damage to persons or property as a matter of products liability, negligence or otherwise, or from any use or operation of any methods, products, instructions, or ideas contained in the material herein. Library of Congress Cataloging-in-Publication Data A catalog record for this book is available from the Library of Congress British Library Cataloguing-in-Publication Data A catalogue record for this book is available from the British Library ISBN 978-0-12-816397-9 For information on all Woodhead publications visit our website at https://www.elsevier.com/books-and-journals

Publisher: Cockle, Charlotte Acquisition Editor: Osborn, Patricia Editorial Project Manager: Kubilis, Michelle Production Project Manager: Sojan P. Pazhayattil Cover Designer: Matthew Limbert Typeset by SPi Global, India

CONTRIBUTORS Pavlovich-Abril Alán  Laboratory of Biopolymers, Research Center for Food and Development, CIAD, A.C. Hermosillo, Sonora, Mexico Amparo Alegría  Nutrition and Food Science Area, Faculty of Pharmacy, University of Valencia, Valencia, Spain Zeynep Altintas  Technical University of Berlin, Berlin, Germany Reyes Barberá  Nutrition and Food Science Area, Faculty of Pharmacy, University of Valencia, Valencia, Spain Maria Bindea  Faculty of Food Science and Technology; Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania Marian Butu  National Institute of Research and Development for Biological Sciences, Bucharest, Romania Lavinia Florina Călinoiu  Faculty of Food Science and Technology; Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania Antonio Cilla  Nutrition and Food Science Area, Faculty of Pharmacy, University of Valencia, Valencia, Spain Anju Dhiman  Department of Pharmaceutical Sciences, M.D. University, Rohtak, India Irene Dini  Pharmacy Department, “Federico II” University, Naples, Italy Rohit Dutt  School of Medical and Allied Sciences, G. D. Goenka University, Gurgaon, India Ana Gabriela Flores-Rueda  Research Group on Functional Foods and Nutraceuticals, Department of Chemical and Biochemical Engineering, National Technological Institute of Mexico-Technological Institute of Durango, Durango, Mexico José Alberto Gallegos-Infante  Research Group on Functional Foods and Nutraceuticals, Department of Chemical and Biochemical Engineering, National Technological Institute of Mexico-Technological Institute of Durango, Durango, Mexico Claudia Ivette Gamboa-Gómez  Biomedical Research Unit, Mexican Institute of Social Security, Durango, Mexico Guadalupe Garcia-Llatas  Nutrition and Food Science Area, Faculty of Pharmacy, University of Valencia, Valencia, Spain

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Vandana Garg  Department of Pharmaceutical Sciences, M.D. University, Rohtak, India Gargi Ghoshal  S.S. Bhatnagar University Institute of Chemical Engineering & Technology, Panjab University, Chandigarh, India Rubén Francisco González-Laredo  Research Group on Functional Foods and Nutraceuticals, Department of Chemical and Biochemical Engineering, National Technological Institute of Mexico-Technological Institute of Durango, Durango, Mexico Jahidul Islam  Laboratory of Nutrition, Graduate School of Agricultural Science, Tohoku University, Japan R. Jayabalan  Food Microbiology and Bioprocess Laboratory, Department of Life Science, National Institute of Technology, Rourkela, India Yearul Kabir  Department of Biochemistry and Molecular Biology, University of Dhaka, Bangladesh Sushil Kumar Kansal  S.S. Bhatnagar University Institute of Chemical Engineering & Technology, Panjab University, Chandigarh, India Kiran  Department of Pharmaceutical Sciences, M.D. University, Rohtak, India María Jesús Lagarda  Nutrition and Food Science Area, Faculty of Pharmacy, University of Valencia, Valencia, Spain Armando M. Martín Ortega  Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Mérida, Mexico Edwin E. Martínez Leo  Unidad de Posgrado e Investigación, Universidad Latino; Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Mérida, Mexico Laura Mitrea  Faculty of Food Science and Technology; Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania Martha Rocío Moreno-Jiménez  Research Group on Functional Foods and Nutraceuticals, Department of Chemical and Biochemical Engineering, National Technological Institute of Mexico-Technological Institute of Durango, Durango, Mexico Onaolapo A.Y.  Department of Anatomy, Faculty of Basic Medical Sciences, Ladoke Akintola University of Technology, Ogbomosho, Nigeria Onaolapo O.J.  Department of Pharmacology and Therapeutics, Faculty of Basic Medical Sciences, Ladoke Akintola University of Technology, Osogbo, Nigeria Semih Otles  Department of Food Engineering, Faculty of Engineering, Ege University, Izmir, Turkey Vasfiye Hazal Ozyurt  Department of Food Engineering, Faculty of Engineering, Near East University, Lefkosa, Turkey

Contributors  xv

Andrea Mariana Păcurar  Faculty of Food Science and Technology; Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania Abigail Meza Peñafiel  Unidad de Posgrado e Investigación, Universidad Latino, Mérida, Mexico Gabriela Precup  Faculty of Food Science and Technology; Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania Cíntia Lacerda Ramos  Basic Science Department, Federal University of Jequitinhonha and Mucuri Valleys, Diamantina, Brazil Nuria Elizabeth Rocha-Guzmán  Research Group on Functional Foods and Nutraceuticals, Department of Chemical and Biochemical Engineering, National Technological Institute of Mexico-Technological Institute of Durango, Durango, Mexico Steliana Rodino  National Institute of Research and Development for Biological Sciences, Bucharest, Romania Rosane Freitas Schwan  Biology Department, Federal University of Lavras, Lavras, Brazil Maira R. Segura Campos  Facultad de Ingeniería Química, Universidad Autónoma de Yucatán, Mérida, Mexico Katalin Szabo  Faculty of Food Science and Technology; Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania Bianca Eugenia Ştefănescu  Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine ClujNapoca;Department of Pharmaceutical Botany, Iuliu Hațieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania Aysu Tolun  Food Engineering, Ankara University, Ankara, Turkey Blanca Denis Vázquez-Cabral  Research Group on Functional Foods and Nutraceuticals, Department of Chemical and Biochemical Engineering, National Technological Institute of Mexico-Technological Institute of Durango, Durango, Mexico Dan Cristian Vodnar  Faculty of Food Science and Technology; Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania Viduranga Y. Waisundara  Australian College of Business & Technology— Kandy Campus, Kandy, Sri Lanka

SERIES PREFACE Food and beverage industry accounts among the most developed sectors, being constantly changing. Even though a basic beverage industry could be found in every area of the globe, particular aspects in beverage production, processing, and consumption are identified in some geographic zones. An impressive progress has recently been observed in both traditional and modern beverage industries and these advances are leading beverages to a new era. Along with the cutting-edge technologies, developed to bring innovation and improve beverage industry, some other human-related changes also have a great impact on the development of such products. Emerging diseases with a high prevalence in the present, as well as a completely different lifestyle of the population in recent years have led to particular needs and preferences in terms of food and beverages. Advances in the production and processing of beverages have allowed for the development of personalized products to serve for a better health of overall population or for a particular class of individuals. Also, recent advances in the management of beverages offer the possibility to decrease any side effects associated with such an important industry, such as decreased pollution rates and improved recycling of all materials involved in beverage design and processing, while providing better quality products. Beverages engineering has emerged in such way that we are now able to obtain specifically designed content beverages, such as nutritive products for children, decreased sugar content juices, energy drinks, and beverages with additionally added health-promoting factors. However, with the immense development of beverage processing technologies and because of their wide versatility, numerous products with questionable quality and unknown health impact have been also produced. Such products, despite their damaging health effect, gained a great success in particular population groups (i.e., children) because of some attractive properties, such as taste, smell, and color. Nonetheless, engineering offered the possibility to obtain not only the innovative beverages but also packaging materials and contamination sensors useful in food and beverages quality and security sectors. Smart materials able to detect contamination or temperature differences which could impact food quality and even pose a hazardous situation for the consumer were recently developed and some are already utilized in packaging and food preservation.

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This 20-volume series has emerged from the need to reveal the current situation in beverage industry and to highlight the progress of the last years, bringing together most recent technological innovations while discussing present and future trends. The series aims to increase awareness of the great variety of new tools developed for traditional and modern beverage products and also to discuss their potential health effects. All volumes are clearly illustrated and contain chapters contributed by highly reputed authors, working in the field of beverage science, engineering, or biotechnology. Manuscripts are designed to provide necessary basic information in order to understand specific processes and novel technologies presented within the thematic volumes. Volume 1, entitled Production and management of beverages, offers a recent perspective regarding the production of main types of alcoholic and nonalcoholic beverages. Current management approaches in traditional and industrial beverages are also dissected within this volume. In Volume 2, Processing and sustainability of beverages, novel information regarding the processing technologies and perspectives for a sustainable beverage industry are given. Third volume, entitled Engineering tools in beverage industry dissects the newest advances made in beverage engineering, highlighting cutting-edge tools and recently developed processes to obtain modern and improved beverages. Volume 4 presents updated information regarding Bottled and packaged waters. In this volume are discussed some wide interest problems, such as drinking water processing and security, contaminants, pollution and quality control of bottled waters, and advances made to obtain innovative water packaging. Volume 5, Fermented beverages, deals with the description of traditional and recent technologies utilized in the industry of fermented beverages, highlighting the high impact of such products on consumer health. Because of their great beneficial effects, fermented products still represent an important industrial and research domain. Volume 6 discusses recent progress in the industry of Nonalcoholic beverages. Teas and functional nonalcoholic beverages, as well as their impact on current beverage industry and traditional medicine are discussed. In Volume 7, entitled Alcoholic beverages, recent tools and technologies in the manufacturing of alcoholic drinks are presented. Updated information is given about traditional and industrial spirits production and examples of current technologies in wine and beer industry are dissected. Volume 8 deals with recent progress made in the field of Caffeinated and cocoa-based beverages. This volume presents the great variety of

Series Preface   xix

such popular products and offers new information regarding recent technologies, safety, and quality aspects as well as their impact on health. Also, recent data regarding the molecular technologies and genetic aspects in coffee useful for the development of high-quality raw materials could be found here. In Volume 9, entitled Milk-based beverages, current status, developments, and consumers trends in milk-related products are discussed. Milk-based products represent an important industry and tools are constantly been developed to fit the versatile preferences of consumers and also nutritional and medical needs. Volume 10, Sports and energy drinks, deals with the recent advances and health impact of sports and energy beverages, which became a flourishing industry in the recent years. In Volume 11, main novelties in the field of Functional and medicinal beverages, as well as perspective of their use for future personalized medicine are given. Volume 12 gives an updated overview regarding Nutrients in beverages. Types, production, intake, and health impact of nutrients in various beverage formulations are dissected through this volume. In Volume 13, advances in the field of Natural beverages are provided, along with their great variety, impact on consumer health, and current and future beverage industry developments. Volume 14, Value-added Ingredients and enrichments of beverages, talks about a relatively recently developed field which is currently widely investigated, namely the food and beverage enrichments. Novel technologies of extraction and production of enrichments, their variety, as well as their impact on product quality and consumers effects are dissected here. Volume 15, Preservatives and preservation approaches in beverages, offers a wide perspective regarding conventional and innovative preservation methods in beverages, as well as main preservatives developed in recent years. In Volume 16, Trends in beverage packaging, the most recent advances in the design of beverage packaging and novel materials designed to promote the content quality and freshness are presented. Volume 17 is entitled Quality control in the beverage industry. In this volume are discussed the newest tools and approaches in quality monitoring and product development in order to obtain advanced beverages. Volume 18, Safety issues in beverage production, presents general aspects in safety control of beverages. Here, the readers can find not only the updated information regarding contaminants and risk factors in beverage production, but also novel tools for accurate detection and control.

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Volume 19, Biotechnological progress and beverage consumption, reveals novel tools used for advanced biotechnology in beverage industry production. Finally, Volume 20 entitled Nanoengineering in the beverage ­industry take the readers into the nanotechnology world, while highlighting important progress made in the field of nanosized materials science aiming to obtain tools for a future beverage industry. This 20-volume series is intended especially for researchers in the field of food and beverages, and also biotechnologists, industrial representatives interested in innovation, academic staff and students in food science, engineering, biology, and chemistry-related fields, pharmacology and medicine, and is a useful and updated resource for any reader interested to find the basics and recent innovations in the most investigated fields in beverage engineering.

Alexandru Mihai Grumezescu Alina Maria Holban

PREFACE In the current era, customers are very demanding for the innovative nutritious foods with enhanced functionality. Diet containing healthy foods and beverages plays a major role in disease prevention and healing of chronic dreading diseases. Functional and medicinal beverages are defined as food products which claim a health benefit. Health-promoting additives found in beverages include proteins, antioxidants, minerals, vitamins, fibers, ω-3 fatty acids, natural plant extracts, prebiotics, probiotics, and other functional ingredients, some of them with unrevealed health beneficial functions. Moreover, current progress made on the concept of modern medicine is increasingly making use of functional and medicinal foods and beverages to heal difficult-to-treat diseases. The scope of this book is to present and discuss the main concepts of functional beverages, novel approaches in their composition, use, and potential health effects. Antioxidant, antimicrobial, anticancer, antiaging and well-being implications of such products are highlighted in this work. The volume contains 15 chapters prepared by outstanding authors from Italy, India, Japan, Romania, Mexico, Turkey, Spain, Nigeria, and Brazil. The selected manuscripts are clearly illustrated and contain accessible information for a wide audience, especially food and beverage scientists, engineers, biotechnologists, biochemists, industrial companies, students and also any reader interested in learning about the most interesting and recent advances in beverage science. Chapter  1, entitled An overview of functional Beverages, by Irene Dini provides an updated review of the various applications of health and wellness drinks, discussing their impact in highly debated current issues such as food intolerance, organic fortified/functional, Better For You Foods, and naturally healthy drinks. Chapter  2, The emerging trends in functional and medicinal beverage research and its health implication, by Gargi Ghoshal et al., discusses the promising trends of usage of functional beverages with special attention on commercial beverages and their health benefits vs their health implications. In the current era, beverages are an ideal delivery vehicle for protein, antioxidants, minerals, vitamins, fibers, ω-3 fatty acids, natural plant extracts, prebiotics, probiotics, and other functional ingredients due to the convenience and prospect to fulfill the customers demand. Nonetheless, there is an increased apprehension over their security in most cases.

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Chapter 3, Herbal extracts—New trends in functional and medicinal beverages, by Steliana Rodino et al., reviews the main knowledge about plant materials, methods of production, biological activities, advantages and risks of using herbal extracts for functional and medicinal beverages. Also, own research results regarding antioxidant and antimicrobial effects of several herbal extracts are shown in this paper. Such herbs include: Equisetum arvense, Humulus lupulus, Tagetes patula, Sambucus spp., Taraxacum officinale, and others. Chapter 4, Bioactive compounds incorporated into functional beverages, by Pavlovich-Abril Alán et al., reviews the latest trends in functional food, attempting to provide a clear definition on what constitute a functional food and the nutriments that have some potentially positive effects on health beyond basic diet, in order to increase the evidence of the beneficial effects of its consumption. In addition, this chapter describes the scientific and technological developments that have the potential of modifying traditional foods and developing new food sources to meet the industrial requirements and the consumer expectations. It also focuses, on the multiple interrelated disciplines approach to examine the health-benefit properties of functional beverage. Chapter  5, Effects and mechanisms of antioxidant rich functional beverages on disease prevention, by Jahidul Islam et al., illustrates the recent studies on the antioxidant and radical scavenging activity of fruit and vegetable juices supplementation and their role in disease prevention, and special attention is paid to the mechanisms of action. Oxidative stress may lead to elevate reactive species and a decrease in antioxidant defenses, resulting in chronic diseases like cardiovascular disease, cancer, type-2 diabetes, and neurodegeneration. Consumption of natural compounds with an antioxidant profile may be a preventive substitute. Chapter 6, Anticancer potential of functional and medicinal beverages, by Vandana Garg et  al., reports on potential health benefits of functional and medicinal beverages related to their consumption as an alternative source of medicine to treat cancer. Functional beverages like tea, coffee, juices, and diverse type of fruit drinks are gaining immense popularity as an alternative source of nourishing human body with desired vitamins, minerals, antioxidant, fatty acids, prebiotics, and probiotics. Chapter  7, Medicinal properties and functional components of beverages, by Aysu Tolun et  al., presents an extensive review of medicinal properties and functional components of beverages with the recent developments in the field. It covers classification of functional beverages with a particular focus on fermented, dairy-based, ­nondairy-based, fruit-based, and herbal-based functional beverages. Furthermore, the significance of bioaccessibility of these products and

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their health ­benefits are discussed. Current in vitro and in vivo studies in the field are also covered. Chapter 8, Development of functional beverages: The case of plant sterol-enriched milk-based fruit beverages, by Antonio Cilla et al., focuses on the development of functional plant sterol-enriched milkbased fruit beverages considering: (i) PS ingredient selection; (ii) bioaccessibility: influence of matrix modification; (iii) bioavailability; and (iv) bioactivity. This integrated approach could serve as an example of the unequivocal demonstration of the efficacy of these functional beverages in promoting health benefits. Chapter 9, Herbal beverages and brain function in health and disease, by Onaolapo et  al., examines the history of herbal beverages, their presumed general health benefits, how their components may affect brain neurotransmitter functions, their potential applications in the management of central nervous system disorders, and adverse effects that may arise from their consumption, especially as it affects the brain. Chapter 10, Functional Beverages from Cereals, by Rosane Freitas Schwan et  al., assesses the relevant findings concerning the characteristics of traditional fermented beverages produced from cereals by focusing on their functional properties. Furthermore, the efforts to improve the health benefits of these beverages will be explored. Chapter 11, Oak leaves as a new potential source for functional beverages: their antioxidant capacity and monomer flavonoid composition, by Nuria Elizabeth Rocha-Guzmán et al., describes some of the health benefits and mechanisms of action of beverages obtained by leaves from several Quercus species, which have proven prophylactic properties from the Mexican traditional medicine. Beverages made of herbal infusions from this source are under development as a new therapeutic tool that may contribute to enhance conventional chemotherapeutic treatments or as a prophylactic consumption alternative. The progress on the biological assays and the challenges of processing these herbal teas are reported. Changes of antioxidant capacity, as well as metabolic profiles of phenolic acids and flavonoids are commented here. Chapter 12, Kombucha as a functional beverage, by Jayabalan et al., discusses kombucha—a beverage which has its origins in the east, while the enzymes, lactic acid bacterial, and other secondary metabolites produced by the microbes during the fermentation qualifies the beverage to be a functional drink. The beverage contains osmophilic yeasts and acetic acid producing bacteria living together symbiotically. Yeasts in particular, convert added sugar in tea to organic acids and ethanol, and acetic acid bacteria uses ethanol to produce cellulose fiber during the fermentation period. As a functional beverage, Kombucha has shown much promise as a drink which can be easily

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prepared in households as well as with the potential to upscale into a commercially produced beverage. Its sensory properties are generally well received by consumers, although areas for improvement such as reducing/modifying its presence of alcohol still exist. Chapter  13, Probıotıc and prebıotıc beverages, by Semih Otles et  al., presents the scientific studies related with probiotic and prebiotic beverages and the potential health benefits related to their consumption. The health effects of the probiotics include alleviation of lactose-­intolerance symptoms, treatment of viral and antibiotic-­ associated diarrhea, reduction of symptoms of antibiotic treatment of Helicobacter pylori, alleviation of atopic dermatitis symptoms in children, and prevention of the risk of allergy in infancy, alleviation of symptoms of inflammable bowel disease (IBD) and irritable bowel syndrome (IBS), and enhancing the immune response. Chapter 14, Probiotics beverages. An alternative treatment for metabolic syndrome, prepared by Martínez Leo et al., aims to review the gut microbiota modifications on people with metabolic syndrome, and discuss the role of probiotic beverages in the chronic inflammatory process, insulin resistance and the microbiome balance reestablishment. Chapter 15, A new generation of probiotic functional beverages using bioactive compounds from agro-industrial waste, by Dan Cristian Vodnar et al., highlights the functional beverage sector as nowadays consumers are more health-conscious and interested in the active role of food to prevent lifestyle diseases which are menacing the wellness of society. The development of new probiotic nondairy functional beverages, with an inflated demand from the lactose-intolerant and vegetarian population is in vogue. Therefore, an emerging trend with respect to designing a probiotic beverage is fortification with selected strains having a positive impact on gastrointestinal health, targeting the irritable bowel syndrome. The presence of bioactive molecules, such as fatty acids and phenolic compounds, in agro-industrial waste makes fruit and vegetable leftovers more valuable for the food industry. Fruits and vegetables contain bioactive compounds that impart health benefits beyond basic nutrition. These wastes are rich in bioactive compounds and can thus be improved and incorporated into food supplements.

Alexandru Mihai Grumezescu University Politehnica of Bucharest, Bucharest, Romania

Alina Maria Holban University of Bucharest, Bucharest, Romania

AN OVERVIEW OF FUNCTIONAL BEVERAGES

1

Irene Dini Pharmacy Department, "Federico II" University, Naples, Italy

1.1 Introduction Today, taste alone is not enough to satisfy consumers, who are looking for high-quality beverages dense in nutrients, therefore ­nonalcoholic beverage companies have been converging on low- or no-calorie drinks and put emphasis on organic ingredients in non­ alcoholic beverage. Functional beverages are an important segment of functional food products since they permit to include desirable ­nutrients and bioactive compounds in order to preserve human hydration and to have antiaging, energy supplying, relaxing, or beauty-­ enhancing effects (Table 1.1) (Corbo et al., 2014). Cinnamon extracts in soft drinks to tackle diabetes or omega-3 s for heart health until ‘beauty beverages’ with collagen, reunited the consumers’ demand for good taste and ingredients that are good for physical fitness and mental well-being. The nanotechnology has made possible to fortify delicate bases, such as water, with whey protein and minerals, causing a minimal effect on the overall taste and mouthfeel.

1.2  Functional Beverage Segment 1.2.1  Energy Drinks Energy drinks are the main segment in functional beverages followed by sports drinks and nutraceutical drinks. They are a group of beverages used by consumers, especially young people who commonly mix energy drinks with alcohol, to provide an extra boost in energy, a cognitive enhancement, to reverse fatigue effects, to maintain alertness, and for endurance. Generally, energy drinks contain caffeine, taurine, glucuronolactone, sugar, B vitamins, and herbal extracts (ginseng, guarana, yerba mate, and green tea extracts), therefore the habitual consumption may enhance: Functional and Medicinal Beverages. https://doi.org/10.1016/B978-0-12-816397-9.00001-7 © 2019 Elsevier Inc. All rights reserved.

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2  Chapter 1  An Overview of Functional Beverages

Table 1.1  Functional Beverages Functional Drink

Ingredients

Energy drink

Taurine, caffeine, and herbal extract such as ginseng, guarana, yerba mate, and green tea extracts Amino acids (GABA or l-theanine) and lemon balm Vitamins, minerals, antioxidants, amino acids and in some cases, fatty acid ingredients in water Protein-enriched drinks beyond weight trainers. Most popular drinks with whey protein isolates, collagen proteins, MCT, etc. Coconut water. It is the perfect sports drink providing you with all the minerals and vitamins you need after your exercise Are very popular to use vitamin blends that contains not only vitamins, but such functional ingredients as Lutein, Omega 3, goji extract, ginseng, etc. Sugar reduced products or naturally sweetened (Stevia, Monk fruit extract or so-called Lo-han, grape juice concentrate, Xylitol, or agave) Soluble dietary fibers (pectin, inulin, gums, and mucilages) Insoluble dietary fibers (cellulose, hemicelluloses, lignin-noncarbohydrate compounds) Goji juice concentrate, acerola, pomegranate Most trendy are those with a Brazilian origin. Acai, maca, camu camu, or Yerba mate and artichoke extracts

Drink and relax Fortified bottled water Proteins for muscles and well aging Simply natural Vitamin premixes Sweeteners Fibers

Juice concentrates Herbal extracts

✓ the risk for caffeine overdose in caffeine abstainers as well as habitual consumers of caffeine from coffee, soft drinks, and tea; ✓ the rate of alcohol-related injury in the case of combined use of caffeine and alcohol; and ✓ alcohol, nicotine, and another drug dependence (Table  1.2) (Abdulrahman, 2015).

1.2.2 Sports Drinks Sports drinks are flavored beverages that are formulated to help people to rehydrate during or after exercise. They are designed to block dehydration and a depletion of the body’s carbohydrate stores. They promote voluntary fluid intake, the emptying of the drink from the stomach, and the quick absorption into the intestine. Sports drinks are developed using essential minerals like sodium, potassium chloride, calcium, pHosphate, and magnesium, which are lost by sweating during exercise; amino acids are able to slow fatigue and improve

Chapter 1  An Overview of Functional Beverages   3

Table 1.2  Adverse Effect due to Consumption of High Levels of Caffeine Susceptible Individuals

Effects

Everyone consume energy drink

Reduce sleepiness Sexual assault and driving while intoxicated (mixing energy drinks) Underestimate true level of impairment (mixing energy drinks with alcohol) Cardiovascular problems Miscarriages, stillbirths, and small for gestational-age infants Elevated blood pressure, anxiety/nervousness, hyperactive behavior, and sleep disturbances

Pregnant women Adolescents

muscle function; B vitamins are used to boost metabolism and generate energy; simple carbohydrates are able to obtain a quick energy burst and complex carbohydrates employed for replenishing energy reserves during and after exercise. There are three main types of sports drinks, according to their osmolality: • Isotonic drinks They contain similar concentrations of salt (46–69 mg/100 mL) and sugar (6–8 g/100 mL) as in the human body. Sodium in the beverage determines the retention of the ingested fluids, stimulate thirst and increase voluntary fluid intake. It can be taken during middle and long-distance running or in team sports, where both the dehydration and the depletion of carbohydrate stores may limit performance. • Hypertonic drinks They contain a higher concentration of sugar (≥10 g/100 mL) than the human body and generally do not contain electrolytes. Hypertonic sports drinks should not be used to maintain hydration as the large amount of carbohydrate is known to slow the gastric emptying and the time for the drink to be absorbed in the intestine. They can be taken post-workout to top-up muscle glycogen stores and on ultra-distance. • Hypotonic drinks They contain a lower concentration of salt ( 70% in the fermented cashew apple juice sample. It was observed that during storage, enhancement of yellow color was observed and higher chroma values from 3.2 to 5.0 after storage for 42  days were presented. Results of sensory study of sweetened fermented beverage after storage period discovered that it was strongly recommended by the panelists with high sensory score. Sun-Waterhouse et al. (2013) studied the exploitation budding of Feijoa fruit wastes as constituent for functional foods. They established that Feijoa fruits have high AOA as they include significant amount of vitamins, PPs, and carotenoids. This study estimated the physicochemical properties like color, pH, total soluble solids (TSS), pectin, fiber content, TEPC, and total AOA of the fruit pomace residual after consumption of Feijoa flesh. Two different extraction temperatures were used such as room temperature and 50°C using hasten solvent extraction method applying water and acidified water of pH 3.5 as extraction media and aqueous ethanol of concentration 30%, 50%, and 80% (v/v). They explained that the characteristics of the extracts mainly dependent on the extraction media and the extraction temperature. A total of 80% (v/v) ethanolic extract produced brighter green in color whereas the more brown extracts were obtained when water and acidified water was used for extraction signifying a probable sensory damaging impacts for using in food. It was found that 50% ethanolic extract had the highest TEPC and AOA done by high-performance liquid chromatography at both extraction temperatures whereas TSS of the extract decreased in the order of 80%, 50%, and 30% concentration of ethanol and water. The produced extracts with lower ethanol

Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH   57

c­ oncentration or water extracts had higher pectin contents whereas water extract produced highest UA content (5.56% as GalA) at 20°C, and also 30% ethanol produced highest GalA (3.90%) at 50°C. They explored that higher extraction temperature (50°C) resulted in lower pectin contents. These results confirmed that the prospects of 50% and 80% ethanolic extracts of feijoa fruit waste can be used as a constituent for the functional food applications. Cassani et al. (2017) studied the combined use of US and vanillin to get excellent attributes and security of strawberry juice supplemented with prebiotic fibers. Prior optimization of preservation techniques using 1.25  mg/mL vanillin and 40 kHz, 180 W US for 7.5 min was used in case of inulin, oligofructose containing strawberry juice. The detection of physicochemical, nutritional properties, microbial quality, and sensory analysis of treated juices was studied. It has been established that no adverse effect was observed on the quality of fresh strawberry juice when inulin and oligofructose was incorporated. Moreover, sensory attributes of the prebiotic incorporated juice was persistent. The applied preservation method improved practically all quality criteria during storing, decreasing microbial growth, particularly lactic acid bacteria (LAB), yeast, and mold growth, which quickly multiply in control juices. Nutritional attributes was also enhanced with increasing amount of total polyphenols and total flavonoid contents and loss of ascorbic acid decreased throughout storage, representing higher antioxidant value. Overall, the estimated sensory attributes of treated juices were believed to be satisfactory (> 2.5). With the incorporation of vanillin pleasant flavor notes were savored for the juice, well matched with the fruit product. Therefore, the performance of treated juice was assessed with respect to postharvest contaminations with concern pathogens of the food industry in terms of health concern taking Listeria ­innocua as the surrogate using Escherichia coli O157:H7 and Listeria ­monocytogenes strain. The optimized treatment resulted in decreased microbial counts during storage reaching untraceable values after 7  days of storage. Thus, the permutation of vanillin content and extend of US treatment could be a practical option to guarantee security and superior quality attributes of strawberry juice supplemented with prebiotic fibers. Carbonell-Capella et  al. (2016) investigated the changes of antioxidant constituent in a fruit juice-stevia rebaudiana mix produced by pulsed electric technologies (PEF) and Ultrasound (USN). In this study papaya and mango juice blends along with sweetener stevia rebaudiana was used. The objective of their study was to assess the effects of the nonthermal emerging technologies on ascorbic acid, total carotenoids, total phenolic compounds, total anthocyanins, and antioxidant capacity of a fruit juice mix. The emerging technologies

58  Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH

used were high-voltage electrical discharges (HVED), PEF, USN, etc. In each technology experimental conditions were designed at two different input energy ranging from 32 to 256 kJ/kg. They applied principal component analysis (PCA) to understand the role of ascorbic acid on total carotenoids and ORAC values. Those factors were very well represented when PEF technology was used. Nonetheless, the use of HVED and USN technologies cannot be excluded, because they may increase the percentage of other functional bioactive compound, for example, in case of total phenolic compounds while HVED technology is useful at an input energy of 256 kJ/kg. The achieved data can be supplied during the optimization of process conditions for manufacturing of highly nutritious beverages.

2.3  Fermented Beverages vs. Nutrition Fruits and vegetables are the rich resource of vitamins e­ specially aqueous soluble vitamins such as C, B-complex, provitamin A, ­minerals, fibers, phytochemicals, and phytosterols beneficial for human health (Gebbers, 2007). As vegetables belong to neutral pH group, low in sugar, and rich in minerals and vitamins therefore vegetables can be used as a common medium for LA fermentation (Buckenhuskes, 1997). LA fermentation improves the organoleptic and nutritional value of the fermented fruits and vegetables and it preserves the vitamins, colored pigments, and other nutrients (Dahal et al., 2005). LA fermentation of vegetable products is one of the safe shelf life enhancement process for the fabrication of partially or completely processed products and is measured as a very important techniques and is furthermore explored due to the rising quantity of raw materials processed in the food industry, and these foods are appropriate to promote the positive health implications of probiotic (Montet et  al., 2006; Heller, 2001). Yamano et al. (2006) explored the utilization of fruits and vegetables fermented by LA facilitates to develop human nourishment in numerous techniques such as the understanding of appropriate nutrition given by that v­ itamins, minerals, and carbohydrates, and avoiding numerous deadening diseases such as diarrhea and cirrhosis of liver due to the presence of probiotic characteristics. According to Kaur and Kapoor (2001), several fermented fruits and vegetables restrain coloring pigments, antioxidants in nature like β-carotene, lycopene, anthocyanins, flavonoids, and glucosinolates they scavenge injurious free radicals, concerned to worsen diseases like aging, arthritis, cancer, etc. Lactobacilli are the extensively studied and most widely used ­probiotics among the LAB. Most Lactobacillus strains belong to the L. salivarius, L. paracasei, L. plantarum, L. reuteri, L. acidophilus group and it indicate the phylogenetic class, are well known to include

Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH   59

­ robiotic organisms. As it is beneficial for well-being of human, it p should accomplish numerous criteria. It must stay alive during passing all the way through the upper GIT and arrive at its location of action lively, and it should be able to survive and act inside the gut atmosphere. The convenient requirements of probiotics offer acceptance to human gastric juice and bile, adherence to epithelial exterior, resolution in the human GIT, stimulation, immunity, and hostile movement toward gut pathogens (such as Helicobacter pylori, Salmonella spp., Listeria monocytogenes, and Clostridium difficile), and the capability to become stable and modulate the intestinal microflora (MattilaSandholm et  al., 2002; Millette et  al., 2008; O’May and Mac Farlane, 2005; Parvez et al., 2006; Anandharaj and Sivasankari, 2014).

2.3.1  Improving Quality of Food and Safety by Fermentation Nutritional attributes of food can be improved by fermentation, which may provide better metabolism, digestion, and absorption and favorable constituent of fermented food. The raw ingredients improve the intensity of vitamin and mineral substances compared to their initial composition. Numerous valuable antimicrobial compounds such as hydrogen peroxide, diacetyls, organic acids, and bacteriocins are formed through the course of fermentation, which hinder the unwanted bacterial growth and therefore increase the shelf life of food. During consumption of fermented food product along with LA it may increase calcium, phosphorus, and iron consumption and also augment adsorption of iron and vitamin D. Diverse types of enzymes are present in fermented foods and all enzymes can contribute in diverse role to improve quality attributes of food. Lactase degrades lactose into galactose in fermented food product. Galactose is an essential constituent of cerebroside which chains brain growth in child. Likewise proteinases formed by LAB can degrade casein into tiny palatable molecules. Fermented foods are an affluent source of globular fats that can be straightforwardly assimilated.

2.3.2  Elimination of Antinutrient Compounds Several fruits and vegetables posses’ contaminant and antinutritional compounds may be in trace. Those components can be eliminated or inactivated by the accomplishment of microbes during the course of fermentation. Foods from plant origin include a sequence of compound, in a group considered as antinutrients that commonly hamper the absorption of various nutrients and in a number of cases may even give noxious or unwanted physiological properties. Those antinutrients consist of oxalate, protease, lectins, compressed t­ annins,

60  Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH

phytic acid, and α-amylase inhibitors. Several pretreatment and ­cooking methods have been shown to possibly reduce the amount of those antinutrients and eventually the unwanted properties left by them. Therefore it may be concluded that the method followed to make the food is consistently essential similar to uniqueness of the food itself. Presently research is focused on recognizing the consequences of numerous antinutritious components before practical consumptions of the fermented food and analyzing their outcomes.

2.3.3  Enhancement of the Human Health Benefits Numerous researches have illustrated the encouraging results of LAB which can alter the gut microflora completely and stop the growth of any other enteric pathogenic organisms. LAB strains also develop the metabolism, develop the immune system, diminish the hazard of colorectal cancer, handle the serum cholesterol concentration, and eradicate the unwanted toxic composite possess in raw or processed food. The common health benefits of LAB may also be explicated.

2.3.4  Inherent Preservation Currently, customers are mostly very aware of the health concern about using food additives. The health benefits of “organic” and “conventional” foods, processing without any externally added chemical preservatives, are fetching additional attraction. Chemical additives have been used generally to inactivate specific microorganisms by stopping their metabolism. For current millennia LAB was found crucial for fermented foods. LAB plays a crucial function for the protection and microbial safety of fermented foods, consequently endorsing the microbial strength to the final products produced during fermentation. Safeguard of foods is the outcome of manufacturing of diacetyl, antifungal compounds, organic acids, carbon dioxide, ethanol, bacteriocins, hydrogen peroxide, and fatty acids, phenyl LA, and antibiotics like reutericyclin (Settanni and Corsetti, 2008). The term “bacteriocin” was first introduced in 1953 to identify colicin formed by E. coli like LAB. Consumption of bacteriocins have also been known for millennia by human being as generated by LAB and, due to that reason, they may be regarded as natural food constituents. Cotter et al. (2005) defined bacteriocin as “bacteriocins can be used to c­ onfer a rudimentary form of innate immunity to foodstuffs.” Bacteriocins are produced ribosomally and extracellularly proteins or low m ­ olecular-mass ­peptides (usually 30–60 amino acids) released and that have bactericidal or bacteriostatic effects on another bacteria, either in the narrow spectrum of similar species or a broad spectrum of across g­ enus

Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH   61

(Settanni and Corsetti, 2008; Cotter et al., 2005; Garneau et al., 2002). Manufacturing of bacteriocin has been found in various genus of bacteria, which are considered “generally recognized as safe” (GRAS), LAB have attracted a great attention in terms of food safety. In fact, LAB bacteriocins benefit from a food grade and this recommends that food scientists permit the growth of desirable flora in fermented foods or inhibit the growth of specific unnecessary (spoilage causing and harmful pathogenic) bacteria in both fermented and nonfermented foods by using a narrow-host range and broad bacteriocins, respectively. According to Aasen et  al. (2003) concerning the use of bacteriocin-generating starter strains in food fermentation, the main problem is associated with the efficacy of in situ antimicrobial that can be negatively predisposed by numerous features, such as the fastening of bacteriocins with the food components (fat or protein particles) and or food additives (e.g., triglyceride oils), inactivation by inhibitors like proteases or other, alteration in solubility and electrical charge, and transform of the cell wrap of the targeted bacteria. The most current food application of bacteriocin includes their fascination to polymeric wrapper, the technology popular as active packaging. Generally bacteriocins possess cationic nature and simply interrelate to Gram-positive bacteria that contain a high amount of anionic lipids in the membrane identifying the pore formation (Settanni and Corsetti, 2008). Tamang et  al. (2009) explained that functional foods are comparable in manifestation to conventional foods but functional foods being eaten as part of the normal diet. On the contrary functional foods, however, they have established physiological reimbursement and can decrease the risk of chronic ailment beyond basic nutritional purpose, counting protection of gut fitness. When a particular food is being cooked or processed using “scientific intelligence” with or without awareness of how or why it is being consumed, then the food is entitled “functional food.” Thus, functional food offers the body with the requisite quantity of nutrients like vitamins, fats, proteins, carbohydrates, etc., compulsory for its healthy endurance. Spirits either plain or aromatized, liqueurs are the essential part of conventional regional cuisine in the Southeast European and Mediterranean region. Particularly herbal spirits and liquors are methodically unlimited and consumed. The aim of the present research was to progress the bioactive profile of traditional herbal liqueur, just to generate an appealing alcoholic beverages flavored with cocoa to consider the possible uses of cocoa raw materials (cocoa beans, ­cocoa nibs, or cocoa liquor). Thus, the polyphenolic profile, sensory properties, and antioxidant profile of herbal liquors substituted by the macerated cocoa-based ingredients were monitored for 6  months, to assess the kinetics of cocoa polyphenols release and optimization

62  Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH

of the mashing method to achieve the paramount cocoa-enriched herbal spirit compared with control herbal liquor without cocoa exhibited considerably a high quantity of polyphenolic antioxidants. Polyphenol content in terms of gallic acid equivalent (GAE/L) was calculated and it was found that polyphenols content in cocoa nibs, cocoa liqueurs, and cocoa bean maximum polyphenols is present in cocoa nibs.

2.4  Fermented Beverage and Their Characteristics Fermentation is a well-known method to make healthy and nutritionally rich with enhanced health beneficial attributes and reduced disease risk. Ample of innovative efforts have been taken to elevate the traditional fermented beverage industry to commercialize the nondairy fermented beverages made from cereals, fruits and vegetables, soya, etc. Those days are not very far when fermented beverages will prolong to be the major components within the functional food cell. Milk-based fermented yogurt-type beverages where kefir is one of the most popular functional fermented dairy products in North America, and Ymer in Denmark and Western Europe. Dairy-based product occupies 43% of the functional beverage market and consists mainly of fermented products reported by Ozer and Kirmaci (2010). In North America and Europe approximately €46 billion is the recent value of the fermented milks and milk product like yogurt market similarly in Asian market value for the same is approximately 77% of the market. Naturally fermented dairy products having consistency like yogurt have been produced throughout the world. Milk-based fermented products may be made either with pasteurized or unpasteurized skimmed milk from diverse source such as cow, goat, camel, sheep, yak, etc. and sometimes from nondairy plant-based coconut milk too. They are prepared with definite starter culture, back slopping, or endorsed naturally to ferment. Even though milk-based fermented beverages mainly contain LAB, the accurate composition of microflora may vary depending on the factors like supply of milk, its composition, and the type of final product to be made, pre treatment of the milk, starter strain, the l­ ocal environment, processing temperature, hygiene, and the type, and handling/treatment of the container used and length of the fermentation. Fermented beverages made from diverse sources popular in different parts of the world are shown in Table 2.2.

Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH   63

Table 2.2  Fermented Beverages Made From Diverse Sources Popular in Different Parts of the World (Marsh et al., 2014) Sl No.

Product

Substrates

Region

Microflora

1.

Amasi

Cow milk

Zimbabwe, Africa

2.

Aryan

Cow milk

Turkey

3.

Garris

Camel milk

Sudan, Africa

4.

Kefir

Cow milk

Caucasian region, Eastern Europe

5.

Kivuguto

Cow milk

Rwanda, Africa

6.

Koumiss/Airag

Horse milk

Russia, Asia

7.

Kumis

Cow milk

South America (Columbia)

8.

Nyarmie

Camel milk

Africa (Ghana)

9.

Rob

Milk (unspecified)

Africa (Sudan)

10.

Suusac

Milk (unspecified)

Africa (Kenya)

Lactococcus (L. lactis), Lactobacillus, Leuconostoc, Enterococcus. Uncharacterized fungal component LAB: Lactobacillus bulgaricus, Streptococcus thermophilus Bacteria: Lactobacillus (Lb. paracasei, Lb. fermentum and Lb. plantarum), Lactococcus, Enterococcus, Leuconostoc. Uncharacterized fungal component Bacteria: Lactococcus, Lactobacillus, Leuconostoc, Acetobacter; Yeast: Naumovozyma, Kluyveromyces, Kazachstania LAB: Leuconostoc (Leu. mesenteroides, Leu. pseudomesenteroides) and L. lactis. Uncharacterized fungal component LAB: Lactobacillus; Yeast: Kluyveromyces, Saccharomyces and Kazachstania Bacteria: Lb. cremoris, L. lactis, Enterococcus (E. faecalis, E. faecium); Yeast: Galactomyces geotrichum, Pichia kudriavzevii, Clavispora lusitaniae, Candida tropicalis LAB: Leu. mesenteroides, Lb. bulgaricus, Lb. helveticus, Lb. lactis, Lactococcus lactis; Yeast: Saccharomyces cerevisiae LAB: Lb. fermentum, Lb. acidophilus, L. lactis, Streptococcus salivarius; Yeast: Saccharomyces cerevisiae, Candida kefyr LAB: Leu. mesenteroides, Lactobacillus (Lb. plantarum, Lb. cruvatus, Lb. salivarius, Lb. Raffinolactis); Yeast: Candida krusei, Geotrichum penicillatum, Rhodotorula mucilaginosa (Continued)

64  Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH

Table 2.2  Fermented Beverages Made From Diverse Sources Popular in Different Parts of the World (Marsh et al., 2014)—cont’d Sl No.

Product

Substrates

Region

Microflora

11.

Shubat

Milk (camel)

China

12. 13.

Amazake Boza

Rice Various (barley, oats, rye, millet, maize, wheat or rice)

Japan Balkans (Turkey, Bulgaria)

14.

Bushera

Sorghum

Millet flour

15.

Koko Sour Water

Cereal (pearl millet)

Africa (Ghana)

16.

Kvass

Russia

17.

Mahewu

18.

Pozol

Rye bread rye and barley malt/flour Maize, sorghum/millet Maize

Bacteria: Lactobacillus (Lb. sakei, Lb. Helveticus, Lb. brevis) Enterococcus (E. faecium, E. faecalis), Leu. lactis and Weissella hellenica; Yeast: Kluyveromyces marxianus, Kazachstania unisporus, and Candida ethanolica Fungi: Aspergillus spp. LAB: Leuconostoc (Leu. paramesenteroides, Leu. sanfranciscensis, Leu. mesenteroides), Lactobacillus (Lb. plantarum, Lb. acidophilus, Lb. fermentum); Yeast: Saccharomyces (S. uvarum, S. cerevisiae), Pichia fermentans, Candida spp. Africa (Uganda) Bacteria: Lactobacillus, Streptococcus, Enterococcus. Uncharacterized fungal component Bacteria: Weissella confusa, Lb. fermentum, Lb. salivarius, Pediococcus spp. Uncharacterized fungal component LAB: Lb. casei, Leu. mesenteroides; Yeast: Saccharomyces cerevisiae

19.

Togwa

Africa (Tanzania)

20.

Hardaliye

Maize flour, finger millet malt Grapes/mustard seeds/cherry leaf

Africa (Zimbabwe)

Unknown

Mexico (Southeast)

Bacteria: L. lactis, Streptococcus suis, Lactobacillus (Lb. plantarum, Lb. casei, Lb. alimentarium, Lb. delbruekii), Bifidobacterium, Enterococcus. Uncharacterized fungal component LAB: Lactobacillus spp.; Yeast: Saccharomyces cerevisiea, Candida spp

Turkey

LAB: Lactobacillus spp. Uncharacterized fungal component

Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH   65

Table 2.2  Fermented Beverages Made From Diverse Sources Popular in Different Parts of the World (Marsh et al., 2014)—cont’d Sl No.

Product

Substrates

Region

Microflora

21.

Kombucha

Tea

China

22.

Water Kefir

Water/sucrose

Mexico

23. 24. 25.

Tari Fenny Kinnauri/ Angoori Chaulli

Date palm Cashew Apple Grapes

Eastern India Goa, India Himachal Pradesh, India India

Worldwide Bacteria: Gluconacetobacter (G. xylinus), Acetobacter, Lactobacillus; Yeast: Zygosaccharomyces, Candida, Hanseniaspora, Torulaspora, Pichia, Dekkera, Saccharomyces Worldwide Bacteria: Lactobacillus (Lb. casei, Lb. hilgardii, Lb. brevis, Lb. plantarum), L. lactis, Leu. mesenteroides, Zymomonas; Yeast: Dekkera (D. anomola, D. bruxellensis), Hanseniaspora (H. valbyensis, H. vineae) Saccharomyces cerevisiae, Lachancea fermentati, Zygosaccharomyces (Z. lentus, Z. florentina) Alcoholic fermentation Alcoholic fermentation Alcoholic fermentation

26. 27

Jack fruit Wine

Dried wild Apricot Jack fruit

India

S. cerevisiae S. cerevisiae

2.5  Health Benefits of Fermented Beverages Kefir and Koumiss are known for their effect on improvement in gastrointestinal health. de Oliveira et al. (2013) stated that kefir especially has shown positive impact on gastrointestinal region, stimulating effect on the immune system, and exhibit anti-inflammatory and anticarcinogenic property although not through medical test. Lactic fermented milk contains compounds not present in regular milk, e.g., exopolysaccharide. Kefiran in kefir is a natural enrichment, including vitamin (e.g., B12 and K2), folate, and riboflavin content. Fermented

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dairy products contain β-galactosidase activity and a reduced lactose content compared to milk, making them suitable for person suffering from lactose intolerance. Majority of the conventional fermented beverages are not investigated satisfactorily. Regardless of this, still people are aware that among these several beverages are health beneficial and mostly in societies where the beverage is occasional in regional custom. They contribute to their selling prospective and justify the spending in associated investigation. Whereas natural fermented milk has effect in controlling hypertension, enhance systematic immunity, lower cholesterol, and lower blood pressure. They can be used to treat IBS and eventually give relief in constipation. Kumar et al. (2012) and Tillisch et al. (2013) demonstrated that they have an anticancer potential and have manipulating effect on intelligence and appropriate for person having lactose intolerance. They are also good resources of bioactive peptides liberated during fermentation through photolytic culture which is related to digestion, endocrine secretion, cardiovascular functions, and the improvement of function of nervous systems enhancement of immunity. The incidence of organic acids that reduce the pH of the beverage may also indicate health beneficial effect. The existence of glucuronic acids in kombucha has detoxification effect by binding with toxin molecule and helps to remove as excreta through urine. They contain B vitamins. It was considered that during fermentation bacteria produce some organic acids, phenols, and antimicrobial agent that have therapeutic, antioxidant, and antimicrobial effects. The cereal-based beverages contain high amount of minerals and have lower fat content than dairy beverages making them nutritious but they lack in essential amino acids. Cereal-based beverages provide plant-based nutrient such as fibers, minerals, vitamins, phenolic compounds, and flavonoids that can affect hyperglycemia, carcinogenesis, inflammation, and oxidative stress. As mentioned earlier fermented foods are mainly common in Africa, they are accustomed to sour taste. Contributing safe, fermented cereal beverages with consistent probiotic cultures could help to reduce diarrhea and malnutrition sources by infected conventional beverages served to weaning children, and facilitate to reduce the extent to the sufferers and help them to recover. Regardless of health claim correlated to the microbial content of fermented beverages and the magnitude in which they survive in this type of beverages and there is a significant deficit in investigation relating to the microorganism attend and the amount in which they survive in such beverages. In order to deal with, it is essential for the purpose of reasonably standard method to evaluate beverages from diverse section to arrive at consent on the justification of microorganisms which comprise part of any specific beverage.

Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH   67

High sequencing and throughput-based microbial laboratory analysis, metabolomics, and bioinformatics look like appropriate technique to provide more appropriate image of these population, triumph problem related to relying on phenotypic-based advancement. Profundity molecular studies have the prospective to be principally helpful while carrying out investigation across different beverages in view to characterize unambiguous advantageous or non-­advantageous sensory and organoleptic characteristics with explicit microorganisms in attendance (Marsh et al., 2013). Such advances will also eventually assist correct species identification, principal to novel starter design, and the development of beverages with different and complex flavor profile. It will also be possible to more effectively monitor the change of proportion of different species throughout the fermentation and storage (Cocolin et al., 2013). Presently commercial Kefir is formed by definite starter with probiotic strains added to some commodities to improve apparent health claims. Nowadays remarkable effort has been taken for the utilization of food industry waste and its value addition whey is one of the valuable, nutrient (55% of milk nutrient and 0.36% fat)-rich by-product of cheese industry and has potential of further use in human diet. Probiotic bacteria can survive in whey therefore probiotic-rich beverage can be made. Prebiotic like oligofructose, inulin, etc., and hydrocolloids can be incorporated to improve viscosity and mouthfeel. Another example is to develop whey-based fruit drinks that have considerable market value and consumer acceptance. Fruit juices are often fortified with vitamins, minerals having high nutrient and antioxidants content and present a new method of nutrient and antioxidant delivery. Tomato, pineapple, pomegranate, orange, cashew, and apple are extensively used to make whey beverages. Probiotic organisms have impact on increasing flavanone and ­carotenoids in orange juice and shown a good survival rate during storage of beverages. Final content of such beverages should be very acidic and best suited to fermentation by probiotic organisms (L. acidophilus, L. plantarum, L. paracasei, L. delbruekii). Microencapsulation technology could aid in the d ­ elivery of other viable microorganisms. Example of commercially available probiotic containing fruit juices include Biola@, Bioprofit@. Analogous microorganisms have also been shown to effectively ferment a variety of vegetable juices like cabbage, beet, pumpkin, courgette, and carrot juices supplemented with ­probiotics. Mix juices, flavor enhancer, natural, or artificial sweeteners can be added to mask the unwanted flavor to improve sensory appeal. Inclusion of bioactive nutraceuticals such as ω-3 fatty acids, isoflavones, and phytosterols can be added. Isoflavones are powerful a­ ntioxidants equivalent to vitamin E and have lowering

68  Chapter 2  THE EMERGING TRENDS IN BEVERAGE RESEARCH

cholesterol e­ ffect. Variety of vitamin and minerals can be added including v­ itamins E, D, and C, calcium, magnesium, iron, especially in fermented milk for preschool children (Silva et al., 2008). It has been shown that taste, price, and base nutritional composition are more important than functional properties.

2.6 Conclusion Nowadays, urbanized civilization turns into more health attentive especially in case of rising obesity problems like an epidemic. The marketplace for functional food products emerges to be extended, defensible trends, for beverages comprising a considerable distribution of the same. Apart from selling to health cognizant and high earning customers, this indicates that functional beverages could act as health beneficial products, mainly as a technique of conveying nourishment, and recovering the healthiness of undernourished group of mass. The therapeutic impression may also be enlarged to emergent cluster of nutraceuticals, accumulation of cholesterol-affecting aspect, for probiotics, the enhancement of stomach ache along with retrieval from antibacterial performance. One feature that must be considered during the advancement of beverages is the necessity to correctly access the marketplace potential in case of a particular product. The apparent difficulty is customers’ eagerness to allow an unusual products and the correct amalgamation of starter and substrate, most favorable nourishment and flavor improvement and systematically sustainable health benefits need to be cautiously measured. Certainly this is the main obstacle in the selling of such goods particularly focusing on the growing alertness along with the customers and the presence of stringent regulations. Since the charges of growth and medical trials and uniqueness in the functional beverage market, there is requirement to have combined attempt among the industry collaborator and academic partner.

References Aasen, I.M., Markussen, S., Møretrø, T., Katla, T., Axelsson, L., Naterstad, K., 2003. Interactions of the bacteriocins sakacin P and nisin with food constituents. Int. J. Food Microbiol. 87 (1–2), 35–43. Anandharaj, M., Sivasankari, B., 2014. Isolation of potential probiotic Lactobacillus oris HMI68 from mother’s milk with cholesterol-reducing property. J. Biosci. Bioeng. 118 (2), 153–159. Bjerrum, K.S., 1996. New applications for pectins. Food Technol. 3, 32–34. Bollinger, H., 2001. Functional drinks with dietary fibre. Fruit Process 12, 252–254. Buckenhuskes, H.J., 1997. Fermented Vegetables. ASM Press, Washington, DC. Carbonell-Capella, J.M., Buniowska, M., Barba, F.J., Grimi, N., Vorobiev, E., Esteve, M.J., Frígola, A., 2016. Changes of antioxidant compounds in a fruit juice-Stevia ­rebaudiana blend processed by pulsed electric technologies and ultrasound. Food Bioprocess Technol. 9, 1159–1168.

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Cassani, L., Tomadoni, B., Ponce, A., Agüero, M.V., Moreira, M.R., 2017. Combined use of ultrasound and vanillin to improve quality parameters and safety of strawberry juice enriched with prebiotic fibers. Food Bioprocess Technol. 10, 1454–1465. Celli, G.B., Ghanem, A., Brooks, M.S., 2015. Bioactive encapsulated powders for functional foods—a review of methods and current limitations. Food Bioprocess Technol. 8, 1825–1837. Cocolin, L., Alessandria, V., Dolci, P., Gorra, R., Rantsiou, K., 2013. Culture independent methods to assess the diversity and dynamics of microbiota during food fermentation. Int. J. Food Microbiol. 167, 29–43. Cotter, P.D., Hill, C., Ross, R.P., 2005. Bacteriocins: developing innate immunity for food. Nat. Rev. Microbiol. 3 (10), 777–788. Dahal, N.R., Karki, T.B., Swamylingappa, B., Li, Q., Gu, G., 2005. Traditional foods and beverages of Nepal—a review. Food Rev. Int. 21 (1), 1–25. de Oliveira, L.A.M., Lemos, M.M.A., Silva, P.R., Soares, R.A., Trajano, S.J., Flosi, P.V.M., 2013. Microbiological, technological and therapeutic properties of kefir: a natural probiotic beverage. Braz. J. Microbiol. 44, 341–349. Dhingra, D., 2012. Recent research on nutraceuticals rich beverages. J. Food Sci. Technol. 49 (3), 255–266. do Carmo, C.S., Pais, R., Simplício, A.L., Mateus, M., Catarina, M.M., 2017. Duarte. Improvement of aroma and shelf-life of non-alcoholic beverages through ­cyclodextrins-limonene inclusion complexes. Food Bioprocess Technol. 10, 1297–1309. Dogan, M., Toker, O.S., Aktar, T., Goksel, M., 2013. Optimization of gum combination in prebiotic instant hot chocolate beverage model system in terms of rheological aspect: mixture design approach. Food Bioprocess Technol. 6, 783–794. Escobedo-Avellaneda, Z., Gutiérrez-Uribe, J., Valdez-Fragoso, A., Antonio Torres, J., Welti-Chanes, J., 2015. High hydrostatic pressure combined with mild temperature for the preservation of comminuted orange: effects on functional compounds and antioxidant activity. Food Bioprocess Technol. 8, 1032–1044. Formica-Oliveira, A.C., Martínez-Hernández, G.B., Aguayo, E., Gómez, P.A., Francisco Artés, F., Francisco Artés-Hernández, F., 2017. A functional smoothie from carrots with induced enhanced phenolic content. Food Bioprocess Technol. 10, 491–502. Garneau, S., Martin, N.I., Vederas, J.C., 2002. Two-peptide bacteriocins produced by lactic acid bacteria. Biochimie 84 (5–6), 577–592. Gebbers, J.O., 2007. Atherosclerosis, cholesterol, nutrition, and statins—a critical review. Ger. Med. Sci. 5, 1–11. Hashim, I.B., Khalil, A.H., Afifi, H.S., 2009. Quality characteristics and consumer acceptance of yogurt fortified with date fibre. J. Dairy Sci. 92 (11), 5403–5407. Hegenbart, S., 1995. Using fibres in beverages. Food Prod. Des. 5 (3), 68–78. Heller, K.J., 2001. Probiotic bacteria in fermented foods: product characteristics and starter organisms. Am. J. Clin. Nutr. 73. Kaur, C., Kapoor, H.C., 2001. Antioxidants in fruits and vegetables—the millennium’s health. Int. J. Food Sci. Technol. 36 (7), 703–725. Komes, D., Belščak-Cvitanović, A., Horžić, D., Drmić, H., Škrabal, S., Borislav, M., 2011. Bioactive and sensory properties of herbal spirit enriched with cocoa (Theobroma cacao L.) polyphenolics. Food Bioprocess Technol. https://doi.org/10.1007/ s11947-011-0630-7. Kumar, M., Verma, V., Nagpal, R., Kumar, A., Behare, P.V., Singh, B., Aggarwal, P.K., 2012. Anticarcinogenic effect of probiotic fermented milk and chlorophyllin on ­aflatoxin-B1-induced liver carcinogenesis in rats. Br. J. Nutr. 107 (7), 1006–1016. Larrauri, J.A., 1999. New approaches in the preparation of high dietary fibre powders from fruit by-products. Trends Food Sci. Technol. 10, 3–8. López-Marcos, M.C., Bailina, C., Viuda-Martos, M., Pérez-Alvarez, J.A., FernándezLópez, J., 2015. Properties of dietary fibers from agroindustrial coproducts as source for fiber-enriched foods. Food Bioprocess Technol. 8, 2400–2408.

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Marsh, A.J., O’Sullivan, O., Hill, C., Paul Ross, R., Cotter, P.D., 2013. Sequencing-based analysis of the bacterial and fungal composition of kefir grains and milks from multiple sources. PLoS ONE 8 (7), 69–71. Marsh, A.J., Hill, C., Paul, R., Paul, R., Cotter, D., 2014. Fermented beverages with health-promoting potential: past and future perspectives. Trends Food Sci. Technol. 38 (2), 113–124. Mattila-Sandholm, T., Myllärinen, P., Crittenden, R., Mogensen, G., Fondén, R., Saarela, M., 2002. Technological challenges for future probiotic foods. Int. Dairy J. 12 (2–3), 173–182. Millette, M., Luquet, F.M., Ruiz, M.T., Lacroix, M., 2008. Characterization of probiotic properties of lactobacillus strains. Dairy Sci. Technol. 88 (6), 695–705. Mishra, S., Mishra, H.N., 2013. Effect of symbiotic interaction of fructo-oligosaccharide and probiotics on the acidification profile, textural and rheological characteristics of fermented soy milk. Food Bioprocess Technol. 6, 3166–3176. Montet, D., Loiseau, G., Zakhia-Rozis, N., 2006. Microbial technology of fermented vegetables. In: Ray, R.C., Ward, O.P. (Eds.), Microbial Biotechnology in Horticulture. Vol. 1. Science Publishers, Enfield, NH, pp. 309–343. Nelson, A.L., 2001. High-fibre ingredients: Eagan press handbook series. Eagan press, St. Paul peroxide-treated lignocellulose from wheat straw. Cereal Chem. 66 (3), 213–217. O’May, G.A., Mac Farlane, G.T., 2005. Health claims associated with probiotics. In: Tamime, A.Y. (Ed.), Probiotic Dairy Products. Blackwell, Oxford, pp. 138–166. Ogundele, O.M.A., Awolu, O.O., Badejo, A.A., Nwachukwu, I.D., Fagbemi, T.N., 2016. Development of functional beverages from blends of Hibiscus sabdariffa extract and selected fruit juices for optimal antioxidant properties. Food Sci. Nutr. 4 (5), 679–685. Ozer, B.H., Kirmaci, H.A., 2010. Functional milks and dairy beverages. Int. J. Dairy Technol. 63, 1–15. Paganga, G., Miller, N., Rice-Evans, C., 1999. The polyphenolic content of fruit and vegetables and their antioxidant activities. What does a serving constitute? Free Radic. Res. 30, 153–162. Parvez, S., Malik, K.A., AhKang, S., Kim, H.Y., 2006. Probiotics and their fermented food products are beneficial for health. J. Appl. Microbiol. 100 (6), 1171–1185. Pereira, A.L.F., Almeida, F.D.L., de Jesus, A.L.T., da Costa, J.M.C., Rodrigues, S., 2013. Storage stability and acceptance of probiotic beverage from cashew apple juice. Food Bioprocess Technol. 6, 3155–3165. Sendra, E., Fayos, P., Lario, Y., Fernandez-Lopez, J., Sayas-Barbera, E., Perez-Alvarez, J., 2008. Incorporation of citrus fibers in fermented milk containing probiotic bacteria. Food Microbiol. 25, 13–21. Settanni, L., Corsetti, A., 2008. Application of bacteriocins in vegetable food biopreservation. Int. J. Food Microbiol. 121 (2), 123–138. Shanmugam, A., Ashokkumar, M., 2015. Characterization of ultrasonically prepared flaxseed oil enriched beverage/carrot juice emulsions and process-induced changes to the functional properties of carrot juice. Food Bioprocess Technol. 8, 1258–1266. Silva, M.R., Dias, G., Ferreira, C.L.L.F., Franceschini, S.C.C., Costa, N.M.B., 2008. Growth of preschool children was improved when fed an iron-fortified fermented milk beverage supplemented with Lactobacillus acidophilus. Nutr. Res. 28 (4), 226–232. Souilem, S., Fki, I., Kobayashi, I., Khalid, N., Neves, M.A., Hiroko Isoda, H., Sayadi, S., Nakajima, M., 2017. Emerging technologies for recovery of value-added components from olive leaves and their applications in food/feed industries. Food Bioprocess Technol. 10, 229–248. Staffolo, M.D., Bertola, N., Martino, M., Bevilacqua, Y.A., 2004. Influence of dietary fibre addition on sensory and rheological properties of yogurt. Int. Dairy J. 14 (3), 263–268.

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Sun-Waterhouse, D., Wang, W., Waterhouse, G.I.N., Wadhwa, S.S., 2013. Utilisation potential of Feijoa fruit wastes as ingredients for functional foods. Food Bioprocess Technol. 6, 3441–3455. Tamang, J.P., Tamang, B., Schillinger, U., Guigas, C., Holzapfel, W.H., 2009. Functional properties of lactic acid bacteria isolated from ethnic fermented vegetables of the Himalayas. Int. J. Food Microbiol. 135 (1), 28–33. Tillisch, K., Labus, J., Kilpatrick, L., Jiang, Z., Stains, J., Ebrat, B., Guyonnet, D., LegrainRaspaud, S., et  al., 2013. Consumption of fermented milk product with probiotic modulates brain activity. Gastroenterology 144 (7), 1394–1401. Uriot, O., Denis, S., Junjua, M., Blanquete-Diot, S., 2017. Streptococcus thermophilus: from yogurt starter to a new promising probiotic candidate. J. Funct. Foods 37, 74–89. Yamano, T., Iino, H., Takada, M., Blum, S., Rochat, F., Fukushima, Y., 2006. Improvement of the human intestinal flora by ingestion of the probiotic strain Lactobacillus johnsonii La. Br. J. Nutr. 95 (2), 303–312. Zare, F., Boye, J.I., Champagne, C.P., Orsat, V., Simpson, B.K., 2013. Probiotic milk supplementation with pea flour: microbial and physical properties. Food Bioprocess Technol. 6, 1321–1331. Zulueta, A., Barba, F.J., Esteve, M.J., Frígola, A., 2013. Changes in quality and nutritional parameters during refrigerated storage of an orange juice–milk beverage treated by equivalent thermal and non-thermal processes for mild pasteurization. Food Bioprocess Technol. 6, 2018–2030.

Further Reading Donoghue, C., Jackson, G., Koop, J.H., Heuven, A.J.M., 2012. Environmental Performance of the European Brewing Sector. Breweries of Europe, Brussels. Paternoster, A., Van Camp, J., Vanlanduit, S., Weeren, A., Springael, J., Braet, J., 2017. The performance of beer packaging: vibration damping and thermal insulation. Food Packag. Shelf Life 11, 91–97. Sun-Waterhouse, D., Bekkour, K., Wadhwa, S.S., Waterhouse, G.I.N., 2014. Rheological and chemical characterization of smoothie beverages containing high concentrations of fibre and polyphenols from apple. Food Bioprocess Technol. 7, 409–423.

HERBAL EXTRACTS—NEW TRENDS IN FUNCTIONAL AND MEDICINAL BEVERAGES

3

Steliana Rodino, Marian Butu National Institute of Research and Development for Biological Sciences, Bucharest, Romania

3.1 Introduction Medicinal and aromatic plants are plant species, also known as medicinal herbs, mainly used for therapeutic, aromatic, and culinary purposes from ancient times. Later, as the knowledge advanced, they began to be used as components in the manufacturing process for cosmetics, medicines, natural foods, and other natural health products. They are also the raw material for processed natural products such as nutraceuticals, food supplements, essential oils for folk medicines, pharmaceutical intermediates, dry and liquid extracts, and oleoresins chemical entities for synthetic drugs. Medicinal and aromatic plants are the main components of global flora biodiversity and reservoirs of bioactive compounds are important (Elisha et al., 2017). Obtaining these bioactive compounds has been a challenge for researchers who have contributed to the development of natural products, to increasing knowledge of the interdependence between the chemical structure of a particular compound and the biological properties of the plant (Atanasov et al., 2015). A large number of plants are known to have antibiotic properties, which is why they have been and are being used intensively in traditional medicine around the world. It is believed that nature gave one remedy (one way or another) of each disease (Dias et al., 2012). In the context of the spectacular evolution of phytotherapeutic use of natural products, the information provided in this chapter was directed to highlight their beneficial properties for human health and the role of plant extracts obtained in different forms (hydroalcoholic, aqueous, essential oils). The demand for medicinal and aromatic plants is constantly increasing due to the industrial development of the production of therapeutic formulations of plants, herbal Functional and Medicinal Beverages. https://doi.org/10.1016/B978-0-12-816397-9.00003-0 © 2019 Elsevier Inc. All rights reserved.

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c­ osmetics, and herbal supplements. Herbal products are more and more prescribed and sold over the counter and the global market of herbal supplements are expected to reach a value of around USD 86.74 billion by 2022, corresponding to an annual growth of 6.8% between 2017 and 2022 (Zion Market Research, 2017). It is estimated that about 3000 species of botanical raw materials are sold around the world. Approximately 350,000 higher plants exist on our planet and related to this number, we could say that only few medicinal plants were studied scientifically. The specialized research groups are trying to fill the information gaps on medicinal plants and herbal extracts, investigating the beneficial effects that their consumption might bring. However, there is scientific evidence of many medicinal plant extracts possessing immunomodulatory, immunostimulatory, antidiabetic, anticarcinogenic, antimicrobial, and antioxidant properties, thus being demonstrated their traditional use in popular medicine.

3.2  Herbal Extracts—Extraction Methods of Bioactive Compounds The first written records on medicinal applications of plants date back to 2600 BCE and report the existence of a sophisticated medicinal system in Mesopotamia, comprising about 1000 plant-derived medicines. Egyptian medicine dates back to about 2900 BCE, but its a most useful preserved record is the “Ebers Papyrus” from about 1550 BCE, containing >700 drugs, mainly of plant origin (Cragg and Newman, 2013). Herbal extracts are an important source of biologically active compounds with a central role in human health. To exert their specific function as a medicinal beverage, the herbal extracts must be carefully prepared, taking into account various parameters. Before the extraction procedure, the medicinal and aromatic plants are usually air dried, in shade, although sometimes the fresh vegetal material may be used. Depending on the plant part used, the material may be cut into pieces, and grinded to a powder (e.g., when using hard pars of the plant, such as rots, stems, rhizomes, or dried fruits) or only gentle crushed or used as such (usually for the softer parts of the plant, such as leaves and flowers). The extraction may be followed by other processes such as filtration, concentration, or drying. The extraction procedure applied is a key element involved in the extraction and identification of the bioactive components of medicinal plants, and not only. Plant-based natural constituents can be derived from any part of the plant like stem, bark, leaves, flowers, roots, rhizomes, fruits, seeds, etc. In practice, various methods of extracting and separating biologically active compounds from plant materials are currently employed.

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The biological activities of plant extracts have shown significant differences depending on the extraction methods applied, thus emphasizing the importance of selecting an appropriate extraction method (Thangaraj, 2016). Along with the traditional and simple conventional methods, several new methods were developed and optimized through the time, but up to this moment no single method is regarded as standard for extracting bioactive compounds from plants. The efficiencies of both conventional and nonconventional extraction methods mostly depend on the critical input parameters (Azmir et al., 2013). Examples of such parameters may be considered: length of the extraction period, solvent used, pH of the solvent, temperature, particle size of the plant tissues, the solvent-to-sample ratio, etc. The beneficial action of herbal extracts may depend on the nature of the plant material, the origin of the vegetal material, the degree of processing, the moisture content, and the particle size. The adjacent technological phases involved in the extraction of medicinal herbs, such as the prior preparation of vegetal material and optimization of the solution obtained, are as well important to be observed: 1. Crushing is accomplished for achieving the rupture of plant organ, tissue, and cell structures such as the constitutive ingredients are better exposed to the solvent chosen for extraction. In other words, this step involves the surface area maximization, thus enhancing the transfer of active compounds from plant material to the solvent. 2. Filtration represents a technological phase performed after extraction. The solution obtained is separated from the vegetal residues (exhausted plant material) by passing it through multiple layers of cheesecloth or specially designed filters, with or without the aid of a vacuum pump. 3. Concentration is performed under vacuum to produce a thick concentrated extract that will be further on dried to produce a solid mass free from solvent. The resulted concentrated solution will be used in a diluted form.

3.2.1 Conventional Extraction Methods Bioactive compounds from vegetal material can be extracted by various conventional extraction techniques, described in detail in the specialized literature. Most of these techniques are based on the extraction power of different solvents and the application of heat and/ or mixing. To obtain bioactive compounds in plants, existing conventional methods, also referred to as solvent-based methods, are considered to be: (1) Infusion (2) Decoction

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(3) Maceration (4) Percolation (5) Soxhlet extraction

3.2.1.1 Infusion Infusion is a method of extraction used to obtain biologically active principles for plant parts that have thinner cell walls—such as flowers, leaves, and thin aerial parts of plants, even some fruit after they have been pre-crushed, containing active thermostable and cold-soluble compounds. For this purpose, boiling or cold water is added to the vegetal material, then this mixture is boiled. The recipient is afterwards covered and left at room temperature for 10–15 min, with occasional shaking. The obtained product is filtered and the result will be dilute solutions of the readily soluble constituents of crude drugs. Infusions are the simplest aqueous extractive solutions obtained from the vegetable raw materials that have especially loose tissues: whole aerial parts of the plant, flowers, leaves, and young stems. The infusions are prepared as needed for up to 12 h. It is kept cold, because extraction in water favors the growth of bacterial mass.

3.2.1.2 Decoction Decoction consists in treating the chopped vegetal material within a specified volume of water (1:4) and boiling it. It is generally recommended for roots, rhizomes, bark, stems, hard fruits, and seeds, that is, those organs of the plant that have a thicker membrane and where the diffusion of the active substances becomes more difficult. Nevertheless, this procedure is also recommended for flowers, leaves, branches, and fruits, especially when extracting volatile oils in the aqueous extract. This procedure is suitable for extracting of ­water-soluble, heat-stable constituents. Usually, the final volume of the extraction solution should be reduced to one-fourth of the original. Also, known under the popular name of boiling, basically, decoction is the liquid obtained by boiling the hard plant parts in water. The extractive solution is filtered hot, the residue is washed with water, and the solution is filled to the initial volume indicated. The decoction solution may be used as such, within 24 h after preparation if kept refrigerated.

3.2.1.3 Maceration Some active substances are better extracted in the cold (heat is precipitated) than by infusion and decoction. Maceration is the simplest extraction method, and is done at the room temperature. The plant product is kept in contact with the solvent (water or water/alcohol mixture) for a varying time, shaking occasionally, after which

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the extract is separated. Cold maceration can be prepared from roots, stems, leaves, flowers, or seeds. This method is best suitable for use in case of the thermolabile compounds. Therefore, maceration consists in treating the chopped vegetable product with the amount of liquid (water or ethyl alcohol 50% or 70%) at a rate of 1:10 g:vol and keeping it at the usual temperature for a certain period of time, shaking from time to time. The final solution is separated by filtration, if possible on a Buckner funnel under vacuum. This extraction method is also used in the preparation of alcoholic extractive solutions or as a preliminary operation in obtaining extracts. For maceration, other solvents than water of alcohol, such as oil, wine, or vinegar depending on the purpose can also be used. A special form of maceration is the production of herbal extracts by digestion. This is a form of maceration in which gentle heat is used during the process of extraction. It is used when moderately elevated temperature is not objectionable.

3.2.1.4 Percolation The percolation method is used most frequently to extract biologically active compounds when the final product takes the form of tinctures and fluidextracts. In the preparation of herbal extracts by percolation, the solvent:solid material ratio, the concentration of the solvent, and the extraction time is usually well established for most of the plant species, and is to be found in Phamacopoea or specialized literature. The most used extraction solvent is 50% or 70% ethyl alcohol, and the usual solvent: vegetal material ratio of 10:1. For the actual extraction, 40–50-mm-percolating columns are used. The first step in preparing the raw material is to moisten the solid ingredients with an appropriate amount of solvent. This mixture is allowed to macerate for a period of time 4–12 h in sealed containers. Then, the solvent is added, and the mixture goes into the percolator columns, and the extraction solvent is passed at a flow rate of 30–50 mL/h. Additional menstruum is added as required, until the percolate measures about three-quarters of the required volume of the finished product (Fig. 3.1). The resultant percolates are concentrated under reduced pressure at a temperature not exceeding 50°C for partial or total removal of the extraction solvent to obtain a hydroalcoholic or aqueous solution. For the complete exhaustion of the plant material, a “battery” of two to three columns can be used.

3.2.1.5  Soxhlet Extraction Hot continuous extraction (Soxhlet) is suitable for large quantities of vegetal material. The raw material is grounded and introduced in the extractor, and after repeated contact between the solvent chosen to dissolve the solvent component, it is enriched in the extracted

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Fig. 3.1  Typical percolation system for the preparation of herbal extracts.

­component (Fig. 3.2). With each wicking, the solvent is enriched in the extract component. After several wrinkles, the heating is stopped, and the vegetal plant is allowed to cool (the water circuit is running during cooling), the cartridge is removed, it is dried and weighed. The main advantage of this method of extraction is that it is accomplished by a continuous process. Generally, the extraction time varies in the Pharmacopeia to a maximum of 30 min for an aqueous extraction and 10 days for an alcoholic extraction. In some cases, the extraction time for aqueous solutions may be extended up to 6 h. Aqueous extractive solutions are generally prepared shortly before use because they have a limited conservation overtime (several hours). The Pharmacopeia provides that 0.1% nipagin may be added to the infusions and decoctions. Also in the Pharmacopeia, it is mentioned that the aqueous extraction solutions can also be prepared by diluting the extracts.

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Fig. 3.2  Typical Soxhlet system for the preparation of herbal extracts.

Regardless of what conventional extraction method is used, it was estimated that the yield of extraction depends mainly on the choice of solvents (Iloki-Assanga et al., 2015). With regard to the preparation of herbal extracts for the production of functional and medicinal beverages, the recommended solvents are water and ethyl alcohol.

3.2.2 Unconventional Extraction Methods Unconventional extraction methods developed over the past 50 years are environmental friendly due to the low volume of synthetic and organic chemicals have a reduced operating time, generate increased yield and superior quality of the final extracts. However, there are both the disadvantage of transferring them on an industrial scale, as well as problems caused by the risk and toxicity to the

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­ perators. Thus, in order to enhance the overall yield and selectivo ity of the bioactive components extracted from the plant material, a variety of techniques have been developed as described in the literature as follows: ultrasound treatment (Chemat et al., 2017), field extraction pulsed electric field (Xue and Farid, 2015; Yan et al., 2017), extrusion, microwave-assisted extraction (MAE) (Karami et al., 2015), and ohmic heating, extraction with supercritical fluids, and extraction with accelerated solvents (Aleksic and Knezevic, 2014). At the same time, the conventional extraction methods such as Soxhlet extraction, percolation, or maceration are still considered to be the reference methods to compare the success of newly developed methodologies, with many papers on this topic (Dhanani et al., 2017; Savoia, 2012).

3.2.2.1  Microwave-Assisted Extraction MAE is a rather recent technique, which utilizes microwave energy to heat the solvent and the probe in order to increase the rate of mass transfer between dissolved substances from the probe matrix and the solvent, contributing to an easier passage in the solvent. The advantage of this technique over the other conventional extraction methods consists of a low extraction time, when a low-energy consumption and solvent is a must and a high-efficiency extraction is required.

3.2.2.2  Ultrasound-Assisted Extraction Ultrasound-assisted extraction (UAE) is one of the most important techniques for the extraction of desirable compounds from the plant material and is fairly adaptable at a small scale or a larger one (e.g., in a laboratory or at industrial scale). UAE is considered as a green method of extraction of compounds of interest, being in the same time an economically viable alternative to conventional techniques. The main benefits of UAE are shorter extraction time, lower amount of energy, smaller quantity of solvents needed, and decreased CO2 emissions (Chemat et al., 2017). Ultrasonic extraction is a modified maceration method, where extraction is facilitated by the use of ultrasounds. The procedure involves the use of ultrasounds, with frequencies from 20 to 2000 kHz, thus increasing the permeability of cell walls and causes their breakage, and favoring the extraction of biologically active compounds. Although the process is useful in some cases, such as root or bark extraction, its large-scale application is limited due to high costs. Another disadvantage of the procedure is known but notoriously harmful but very well-known effect of ultrasound energy (over 20 kHz), active elements of medicinal plants, the formation of free radicals, and consequently undesirable changes in their molecules.

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Therefore, extraction represents the activity of separating the biological active compounds from vegetal material using selective solvents. The final solutions obtained contain a complex mixture of metabolites, in liquid or semisolid state or in dry powder form, and are intended for oral or external use. Water is the universal solvent chosen for herbal extracts, and may be easily used for homemade herbal preparations, such as infusions, decoctions, and macerations. It is usually used for the extraction of constituents with antimicrobial activity. However, water-soluble flavonoids (mostly anthocyanins) have no or very low antimicrobial significance, while only water-soluble phenolics are implicated in the possible antioxidant activity of several plant extracts.

3.3  Biologically Active Compounds From Plants During the last decades, there are various reports showing e­ vidence that plants are rich sources of different classes of antimicrobial substances acting as defense systems to protect themselves against biotic (living) and abiotic (nonliving) stresses (Nabavi et al., 2015b). It is generally known that plants produce a variety of chemical compounds that can be divided into primary and secondary metabolites. Primary metabolites are those compounds that are essential for the emergence, development, and survival of the plant. They include carbohydrates, proteins, and lipids. Secondary metabolites were initially considered to be residual products. They are not essential to the plant, but in the long run their lack can influence survival (Atanasov et al., 2015). It has been shown that secondary metabolites play an important role in adapting plants to the living environment, while at the same time being an important source of natural ingredients for pharmaceuticals. Secondary metabolites, known as phytochemicals, natural products, or biologically active compounds of plants, are responsible for the medicinal properties of the plants in which they are incorporated. Among these secondary metabolites, polyphenols, terpenoids, alkaloids, lectins, polypeptides, and polyacetylenes are known to be the antimicrobial agents; most of these metabolites are also approved as a generally recognized as safe (GRAS) material for food products, showing negligible side effects (Nabavi et al., 2015a). The role they play in plant survival is not, until now, well known or understood, but is definitely more than a protective role (Kabera et al., 2014). The classification of secondary metabolites that can be extracted from plants is based on their chemical structure, composition, solubility in different solvents, or the pathway on which they are synthesized. The main classification system includes three major groups: terpenoids,

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Fig. 3.3  Classification of plant secondary metabolites.

alkaloids, and phenolic compounds, joined by other groups such as amines and glycosides. Each of these groups is divided into subclasses with a complex structure (Hassan et al., 2017; Kasote et al., 2015). Thus, the main biologically active compounds present in plants and which are investigated for possible medicinal properties can be classified according to the schematic representation in Fig. 3.3.

3.3.1 Terpenoids Terpenoids are molecules found in all plants. They are composed of isoprene units. Terpenoids (isoprenoids) are the largest and most diversified class of chemical compounds among plant-derived secondary metabolites, with >40,000 structures identified so far (Bohlmann and Keeling, 2008; Singh and Sharma, 2015). Traditionally, plant-derived terpenoids have been used by people in the food, pharmaceutical, and chemical industries and, more recently, have been exploited in the development of biofuels (Tholl, 2015). Terpenoids are classified according to the number of isoprene units they contain, as follows: isoprene, which is synthesized and released by the plants, comprises one unit and is classified as a hemiterpene. There are also monoterpenes, sesquiterpene, diterpenes, triterpene, and tetraterpene (Goto et al., 2010).

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Volatile oils give plants their perfume. Some plants use volatile compounds to discourage herbivores and protect the plant from certain pests. Essential oils are widely used in aromatherapy and medicine. In aromatherapy, essential oils are considered to improve the mood and mental functioning. In alternative medicine, essential oils are considered to have multiple benefits. In the scientific literature, it has been reported that essential oils derived from aromatic herbs (e.g., fennel, mint, thyme, lavender, etc.) have antimicrobial action against Grampositive and Gram-negative bacteria and yeasts, fungi, and viruses. These oils contain mixtures of volatile substances such as monoterpenes, sesquiterpene, and/or phenylpropanoids (Savoia, 2012).

3.3.2 Polyphenols Polyphenols are one of the most important and at the same time the most numerous of the secondary metabolite groups present in the plant kingdom. Phenolic compounds are virtually omnipresent in the plant kingdom. At present, over 8000 phenolic structures have been identified in a wide variety of forms, of which >4000 belong to the flavonoid class, and of these several hundred are present in edible plants (Gao and Hu, 2010; Wang et al., 2017). Research on vegetal material rich in polyphenols has received considerable attention, on the one hand, on the need to develop innovative drugs for the treatment, prevention, and control of various microbial infections and, on the other hand, for use in industry food, and preservatives to improve the quality and nutritional value of food. Among the polyphenols, a series of pigments with the quinonic structure that are responsible for the color of fruits and flowers (alizarin, purpurin, benzoquinone, juglona) are included. Phenolic compounds are very important for plants and can have multiple functions. These molecules are generally involved in the defense against ultraviolet radiation, oxidizing agents, or the aggression of some phytopathogenic agents (Cowan, 1999) having a role in adapting to biotic and abiotic stress. Polyphenols can be classified into different groups according to the number of phenolic rings they contain and the structural elements linking these rings (Saxena et al., 2012). They are classified into phenolic acids, flavonoids (flavonones, flavones, xanthones, catechins, anthocyanins, anthocyanidins), lignans, stilbenes, and other polyphenols with non-flavonoid structure (Fig. 3.1). Plant phenols include various molecules, ranging from simple, low-molecular-weight compounds to complex polyphenols. The main phenolic compounds in plants can be classified into two groups: flavonoids (polyphenolic compounds formed from 15 carbon atoms and 2 aromatic rings with a 3‑carbon bond) and non-flavonoids (phenolic acids, tannins, stilbene, etc.). Scientific studies have highlighted

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various and important biological activities of phenolic compounds in plants, such as antimicrobial, allelopathic, antioxidant, anti-­ inflammatory, cardioprotective, antimutagenic, anticarcinogenic, and antiaging activities. They also contribute to the induction of apoptosis in various ways, such as cell cycle arrest, migration, proliferation or differentiation, regulation of carcinogenic metabolism, inhibition of cell adhesion, signaling pathways blockade, and others. Phenolic acids are phenols possessing a carboxylic functional group, varying due to the hydroxylation or aromatic nucleus methoxylation. Gallic acid and vanilic acid are present in almost all plants. Caffeic acid is considered to be the most common phenolic compound distributed in the plant kingdom, followed by chlorogenic acid, which is known to cause allergic dermatitis (Marchese et  al., 2014; Saxena et al., 2012). Phenols are essentially a series of natural antioxidants, used as nutraceuticals. However, it is assumed that the total content of polyphenols in plants is underestimated and many of the phenolic compounds and their derivatives have not yet been identified due to limitations of the methods and techniques of analysis used. Along the history, several major groups of plant products with antioxidant and anti-inflammatory capacity have been identified. A particularly valuable class of health-enhancing plant compounds is flavonoids. These are polyphenolic molecules with properties that include free radical scavenging, inhibition of hydrolytic and oxidative enzymes, anti-inflammatory action, reduction of blood-lipids and glucose, and the enhancement of human immunity (Iloki-Assanga et al., 2015). The most common group of plant phenols are flavonoids, the structure from which these molecules are formed, consisting of two phenolic rings linked by a hydrogenated piranic heterocyclic ring. Flavonides or flavon derivatives are an important group of polyphenols widely distributed in the plant kingdom. Flavonoids are structures that are abundant in cells that produce photosynthesis. The main sources of flavonoids in the diet are fruits and vegetables. They are found in common edible plants such as fruit, vegetables, nuts, and seeds. They occur also in certain grains, seeds, and spices, as well as in wine, tea, coffee, cocoa, and herbal essences. Up to now, 14 classes of flavonoids have been identified. These differ depending on the chemical nature and position of the substituents on the different rings (Savoia, 2012). The common classes of flavonoids are flavones, flavonols, isoflavones, anthocyans, catechins (flavanols), and flavanones. There are over 4000 flavonoid identifiable (Amjad and Shafighi, 2013), most of them being pigments of superior plants. Quercetin, kaempferol, and quercitrine are common flavonoids found almost in most plants. Other flavonoids are: flavones (e.g., apigenin or apigenol), flavonols (quercetin and rutoside), calcone, catechins, etc.

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For the identification of flavonoids, the characteristic reaction of cyanidine or the Shibata reaction (treatment of the hydroalcoholic solution of flavonoids with magnesium in the hydrochloric acid medium) is used. Different colors are obtained, namely orange for flavones, red for flavonols, and red-violet for flavanones. A wide range of antioxidant and anti-inflammatory flavonoids are described and identified in the literature.

3.3.3 Alkaloids Alkaloids are natural substances of plant origin, heterocyclic compounds containing nitrogen in the molecule. They have an alkaline reaction and have strong pharmacodynamic properties, even in low doses (Simion et al., 2009). Most alkaloids are solid, such as atropine, or liquids, those containing carbon, hydrogen, and nitrogen. Most alkaloids are readily soluble in alcohol, and although they are poorly soluble in water, their salts are usually soluble. The ­alkaloid solutions are highly bitter. These nitrogen compounds function in protecting plants against herbivores and pathogens and are widely exploited as pharmaceuticals, stimulants, narcotics, and poisons because of their strong biological activity. In nature, there are alkaloids located in larger proportions in certain plant organs, seeds, leaves, peel, and roots. Alkaloids have pharmacological applications as anesthetics and stimulants of the central nervous system (Madziga, 2010). The first useful medical alkaloid was morphine, isolated in 1805 from poppy seed, Papaver somniferum (Cowan, 1999). Solamargine, a glycaalcaloid from Solanum khasianum beans, may be useful against HIV infection as well as other AIDS-related intestinal infections. The identification of alkaloids is accomplished by precipitation reactions with the general reagents containing metals or metalloids: mercury, bismuth, tungsten, and iodine (Table 3.1). The composition of plants regarding polyphenolic substances is unique to a particular species, even depending on subspecies. Moreover, this composition may also differ with respect to the geographical area and the pedoclimatic conditions. Generally, biosynthesis of secondary metabolites is a complex process that is influenced by a number of stress factors such as temperature, environmental pH, microelements, biological interactions, and the presence of other chemical agents. The composition of plants has been of particular interest to scientists, and as the methods of investigation have advanced and become more complex and complete, the subject has been repeated by many “masters” in the field, renowned specialists who have formed schools in this direction and their disciples.

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Table 3.1  Methods for Identifying the Alkaloids Reagent

Chemical Composition

Precipitate Properties

BOUCHARDAT DRAGENDORFF

Aqueous iodinated iodine solution Potassium tetraiodobombed aqueous solution

MAYER-VELTZER

Aqueous solution of biphasic ­tetrahydrate-buffered saline with chloral hydrate Aqueous silicotungstic acid solution

Brown-red Red-orange or orange-yellow, soluble in alcohol, ether and other solvents White, becomes yellowish, crystalline or microcrystalline, soluble in alcohol White, the alkaloid is released from the complex with alkali hydroxides Yellow-red in the presence of dilute sulfuric acid White

BERTRAND MARME SCHULTZE HAGER KNORR IONESCU-MATIU

Aqueous solution of double potassium iodide and cadmium Phosphoantitimonic acid in the presence of phosphoric acid or concentrated sulfuric acid Picric acid 1% Pycrolonic acid 2% Saturated picric acid solution in 5% alcohol, with glycerol

Yellow, soluble in hot environment Yellow or red, precipitate, decomposes in hot environment Yellow crystals

The antibacterial potential of secondary plant metabolites (phytochemicals) materialized in the form of herbal extracts was already demonstrated, both alone and as synergists or potentiators along with other antibacterial agents. The use of herbal extracts as a source of antimicrobial agents is an increasingly active subject of research worldwide. Of the plants listed in popular medicine, particular attention is paid to the use of plants rich in phenolic substances. Tannins are water-­ soluble phenolic substances of particular importance. Extracts from tannin-rich plants have been used in traditional human medicine to treat infectious diseases.

3.4  Herbal Extracts—Trends in Modern Medicine Natural nutraceuticals derived from plant extracts (also named as herbal extracts or botanicals) are used as adjuvants in preventing various diseases and maintaining the healthy state of the human ­organism. Overtime, it was already demonstrated the value of medicinal plants as a source of molecules with therapeutic potential. In the

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Fig. 3.4  Possible pharmaceutical variations of herbal tinctures.

­ resent, they still represent a critical source for the identification of p novel drug precursors (Atanasov et al., 2015). The herbal extracts used for medicinal purposes are based on the healing power of plants, used as such in the form of raw herbal extracts, or essences, powders, and tinctures. As it was already described in the previous section, the herbal extracts with practical application as functional or medicinal beverages can take simple or complex forms (Fig. 3.4). Plants act on pathologies or symptoms, according to the allopathic principles. Conventional, modern medicine uses drugs based on the herbal extracts (e.g., aspirin has been synthesized in the past from the bark of willow tree). Unfortunately, there is a huge industry of the socalled over the counter (OTC) plant-based drugs, sold without prescription. Usually, these natural remedies, including different forms of herbal extracts, are used without a medical prescription, but based on social media or other informal channels, most of which have no scientific evidence. On the other hand, there is an ancient traditional medicine based on old concepts on human health and treatment of ailments. This type of medicine addresses a general state of health, using a combination of remedies targeted on a specific person or patient. Phytotherapy represents all the possibilities of preventing and treating diseases by using products obtained exclusively from plants, such as herbal extracts. Relieving pains, stopping bleeding, and wound healing following the use of natural remedies belongs to the first empirical knowledge. Overtime, this knowledge has begun to be recorded, and with typographic multiplication, they have become ­assets

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of the whole society. The first organized form of the study of herbal medicines was established in the 1500s in Padova. Phytotherapy is basically based on two types of documentary sources: first records, archeological vestiges and works of art that have immortalized the use of plants as medicinal beverages, and on the other hand traditional medicine nowadays. The latter ensures the reconstitution of many archaic prescriptions of medication. Popular medicine always had a special, universal character, and at the same time specialized in the regions. This is due to the use of two types of medicinal herbs − on the one hand, old species, well known, which have penetrated even in cult medicine, doctrine based on scientific evidence (e.g., Equisetum arvense, Hypericum perforatum, Sambucus nigra, Achillea millefolium, and Chamomilla matriciaria) and − on the other hand, species with ethnobotanic attributes specific to the geographical area. In our days, folk medicine is not so much a value in itself, but acts as a reliable source of inspiration for experimental research, as will be demonstrated in the following sections. Self-medication is already a habit overpassed by the progress of science. The folk remedy that has survived since ancient times to reach the present days is likely to become a scientifically recognized drug. The condition is to achieve pharmacognostic studies, with regard to the chemical composition (biologically active substances) their mechanism of action, the beneficial effect they bring on human health, the therapeutic dose, their eventual toxicity. Moreover, the treatment should always be based on a clear diagnosis and mandatory to be prescribed by a doctor. The modern medicine based on natural remedies, such as herbal extracts, aims to provide evidence-based scientific knowledge, validating curative properties, and clinically observing the therapeutic effect. Phytotherapy research is mostly using the data provided by traditional medicine, but it does not exclude the discovery of new pathways.

3.4.1  Herbal Extracts for Cardiovascular Disorders Herbal extracts have been used for congestive heart failure, systolic hypertension, angina pectoris, atherosclerosis, cerebral insufficiency, venous insufficiency, and arrhythmia since centuries. At this moment, the increasing popularity of natural products has revived interest in traditional remedies for the treatment of cardiovascular diseases (Rastogi et al., 2016). A literature review was carried out on the plants that were most frequently cited by a research group based at the University of Navarra, Spain. Official sources [European Scientific Cooperative on Phytotherapy (ESCOP), German Commission E, World Health Organization (WHO), European Medicines Agency (EMA), European

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Pharmacopeia (pH. Eur.), and Real Farmacopea Española] were consulted in order to establish the therapeutic efficacy of the reported uses and to obtain further details about quality and safety aspects. In the same time, information was also collected using interviews with 667 respondents from different locations. Over 450 pharmaceutical uses were reported by the respondents. The most frequently used parts of the plants were the aerial parts followed by leaves and flowers. The authors estimated that 19% of the 90 reported plant species, corresponding to 45% of the pharmaceutical uses had already been pharmacologically validated for their therapeutic efficacy and safety aspects. Eventually, the author’s proposal was the validation of only five species for their use in cardiovascular diseases, as follows: Rhamnus alaternus L. (Italian buckthorn or Mediterranean buckthorn), Potentilla reptans L. (European cinquefoil or creeping tormenti), Equisetum telmateia Ehrh. (great horsetail or northern giant horsetail), Centaurium erythraea Rafn (common centaury), and Parietaria judaica L. (spreading pellitory) (Calvo and Cavero, 2014). Another group reviewed the scientific literature, aiming to provide updated and comprehensive information on the history and traditional uses of herbal extracts for disorders of the cardiovascular system. The electronic literature search was performed using specialized platforms for journals (Pubmed, SciFinder, Scirus, GoogleScholar, JCCC@INSTIRC, and Web of Science) with no restrictions on language of publication. The results showed the possible cardiovascular effects of four plants: Allium sativum (garlic), Commiphora wightii (Guggul), Crataegus oxyacantha (Hawthorn), and Terminalia arjuna (Arjuna). Authors considered that there is enough evidence of these herbal extracts efficacy in various cardiovascular disorders, including ischemic heart disease, congestive heart failure, arrhythmias, and hypertension, although the mechanisms of action are not very clear. However, the potential synergistic and possible adverse side effects of herbal extracts use should be studied, before establishing them as remedies for cardiovascular diseases (Rastogi et al., 2016). More recent, the available evidence based on randomized controlled trials (RCTs) of polyphenol-based supplements was overviewed. The polyphenol-based nutraceuticals and functional foods might be indeed used as adjunct therapy of cardiovascular disease, but additional long-term RCTs with adequate numerosity and with clinically relevant end points are needed to provide unequivocal evidence of their clinical usefulness (Fig.  3.5) (Tome-Carneiro and Visioli, 2016). Pharmacological studies have revealed that several species of plants are effective in cardiovascular health with various mechanisms.

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Fig. 3.5  Evidence-based use of nutraceuticals for cardiovascular disorders. From Tome-Carneiro, J., Visioli, F., 2016. Polyphenol-based nutraceuticals for the prevention and treatment of cardiovascular disease: review of human evidence. Phytomedicine 23, 1145–1174.

Among them, Cinnamomum cassia (L.) J. Presl, Crocus sativus L., Elettaria cardamomum (L.) Maton, Ocimum basilicum L., Melissa officinalis L., Phyllanthus emblica L., and Punica granatum L. were the most efficient (Sobhani et al., 2017). According to the literature survey of Walden and Tomlinson (2011), the most studied herbal extracts of cardiovascular disorders, for which there is some evidence, if not the final proof are as follows: Hawthorn (Crataegus species), Garlic (A. sativum), Danshen (Salvia miltiorrhiza), Lingzhi (Ganoderma lucidum), Maidenhair Tree (Ginkgo biloba), Foxglove (Digitalis purpurea/lanata), and Ginseng (Panax species). However, a systematic review of ginseng practical applications in the prevention and treatment of cardiovascular disorders concluded that there is an insufficient evidence for this herb’s efficacy (Buettner et al., 2006). Although having a long history of use in traditional medicine and showing promising in vitro biological actions, herbal extracts remain clinically unproved for the treatment or prevention of cardiovascular disorders, and are yet often insufficiently standardized to be recommended as therapy and be included in mainstream of healthcare system. The evidence supporting the use of herbal extracts from c­ linical trials is not yet secure, but custom and practice make it likely that they will continue to be used for the prevention or treatment of cardiovascular diseases, together with other indications (Walden and Tomlinson, 2011).

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3.4.2  Herbal Extracts for Digestive System Disorders Herbal extracts are successfully used in chronic disorders of the digestive system, for stomach, liver and biliary tract, colitis, and chronic constipation. Whether it is a simple infusion (tea) obtained from plant mixtures specific to these diseases, be it pharmaceutical products obtained by the most modern technologies available. Not only the chronic diseases of the digestive system and but also for a number of acute cases of symptomatic treatment were achieved very good results. Herbal extracts used in acute stomach and intestinal disorders are characterized by rapid action especially in spastic states, in gastralgia, intestinal colic, or acute dyspepsia. The herbal extracts mostly used for acute stomach and intestinal disorders are: Mint (Mentha piperita), Lemon balm (M. officinalis), chamomile (Matricaria chamomilla L.), caraway (Carum carvi), and butterbur (Petasites hybridus). The medicinal plant species are commonly used for medicinal purposes against digestive disorders out of which most frequently occurring are: stomach ache, diarrhea, indigestion, constipation and inflammation, etc. In a recent study on traditional knowledge and cultural drivers of Manoor Valley, Northern Pakistan, herbaceous plant species are the dominant among plants studied which were 64% of the total plants, followed by trees (20%) and shrubs (16%). Lamiaceae was the leading family among collected medicinal plant species (13.6%). Maximum medicinal plant species were used for the treatment of stomach ache (11.7%), diarrhea, and indigestion (10.9% each) (Rahman et al., 2016). Because Escherichia coli plays an important role in causing diarrhea, it is one of the most investigated microorganism for testing the possible antibacterial activity of various herbal extracts. Diarrhea is a major worldwide health problem, especially among children. Infectious diarrhea is one of the main causes of disease and mortality in the developing countries and is one of the most prevalent causes of mortality in children (Bahmani et  al., 2014). However, when performing experimental studies, the herbal extracts are tested for their antimicrobial activity against a wide range of Gram-positive and Gramnegative bacteria, as well as fungal pathogens. Most of the extracts possess in vitro effect against more than one microbial strain. Therefore, in the following paragraphs the antimicrobial effect of herbal extracts against several bacterial strains will be discussed. An emphasis will be given to bacteria involved in gastrointestinal disorders. In a very recent study, herbal extracts from leaves of nine medicinal plant species with previously demonstrated antibacterial activity against E. coli were evaluated for their efficacy against Gram-positive

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(Staphylococcus aureus, Enterococcus faecalis, and Bacillus cereus) and Gram-negative (E. coli, Salmonella Typhimurium, and Pseudomonas aeruginosa) bacteria. The herbal extracts showed antibacterial activity against all the bacterial pathogens taken into study. Cremaspora triflora and Maesa lanceolata herbal extracts showed the highest antibacterial activity. Some of the herbal extracts had relatively low cytotoxicity with LC50 > 20 μg/mL, while others (M. lanceolata, Elaeodendron croceum, and Calpurnia aurea) had overcome this value. However, the authors pointed that the further investigation is in progress on C. triflora and Hypericum roeperianum, both of which had promising activities and potential safety based on cytotoxicity (Elisha et al., 2017). Al Mariri et al. observed in vitro the antibacterial effect of 28 plant extracts and oils against bacterial strains E. coli O157:H7, Yersinia enterocolitica O9, Proteus spp., and Klebsiella pneumoniae. Among the evaluated herbal extracts, only Laurus nobilis showed antibacterial activity. With regard to the essential oils tested, Origanum syriacum L., Thymus syriacus Boiss., Syzygium aromaticum L., and Cinnamomum zeylanicum L. essential oils were the most effective, leading to conclusion that these oils could act as bactericidal agents against the selected bacterial strains (Al-Mariri and Safi, 2014). Helicobacter pylori is classified as a Gram-negative, spiral, and microaerophilic bacterium that specifically colonizes the gastric mucosa and affects more than half of the world’s population (Bonifácio et al., 2014). H. pylori infection is usually acquired in the early childhood and it can persist throughout the life without antibiotic treatment, affecting about 20% of the population in the developed countries and >90% in the developing world (Calik et al., 2016). Various herbal extracts (including partial purified fractions and isolated compounds) were investigated for their anti-H. pylori effect. In  vitro experiments highlighted a strong anti-H. pylori activity of herbal extracts, some of them being almost equal to antibiotic controls, usually used in the mainstream healthcare system. Further on, the in vivo investigations on animals, revealed than the effectiveness of the plant products decreased H. pylori colonization in the stomach (Wang, 2014). The ethanol extracts of Bixa orellana L. (Bixaceae) seed, Chamomilla recutita L. (Asteraceae) inflorescence, Ilex paraguariensis A. (Aquifoliaceae) leaves, Malva sylvestris L. (Malvaceae) inflorescence and leaves, Plantago major L. (Plantaginaceae) above ground parts, and Rheum rhaponticum L. (Polygonaceae) root exhibited significant activity against the in vitro growth of H. pylori (Cogo et al., 2010). H. pylori eradication with antibiotic regimens has a limitation mostly due to antibiotic resistance. Medicinal plant compounds and other natural products provide another choice or opportunity to eradicate H. pylori infection. However, potential cytotoxicity and adverse

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side effects might present from those medicinal plant products (Wang, 2014). Yesilada et al. have reported that aqueous and methanolic extracts from the Sambucus ebulus aerial parts presented an inhibitory activity against the H. pylori (Yesilada et al., 2014). Min-Chi Lu et  al. determined whether three types of extracts (aqueous extract, ethanol extract, and chloroform extract) obtained from Ixeris chinensis (Thunb.) Nakai (Taiwan) present antioxidative, anti-inflammatory and anti-adhesive effect on H. pylori-induced gastric adenocarcinoma in an adenocarcinoma gastric cell line (AGS) cell model. I. chinensis is a traditional herbal medicine used for treating stomach ache, common cold, and diarrhea. The aqueous extract showed highest antioxidative activity while the chloroform resulted in maximum antibacterial [minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC)], anti-adhesive (urease test), and anti-inflammatory [interleukin-8 (IL-8), nitric oxide (NO), tumor necrosis factor alpha (TNF-α)] activities (Lu et al., 2018). Espinosa-Rivera et  al. assessed five different polarity herbal extracts from roots and aerial parts of Parthenium hysterophorus against H. pylori growth by the broth dilution method. P. hysterophorus is a medicinal plant used to treat gastrointestinal disorders, such as gastritis. The organic extracts inhibited in vitro growth of H. pylori. The dichloromethane extract showed a MIC of 15.6 μg/mL while the aqueous extracts showed low or null activity. All five herbal extracts tested inhibited adherence in different ranges but the dichloromethane-­ methanol ones possessed the highest efficiency, with a 70% maximal inhibition at 1 mg/mL. The results provided by the authors highlighted the antimicrobial activity of P. hysterophorus extracts, that could act synergistically against H. pylori (Espinosa-Rivero et al., 2015). The antibacterial activity of Anisomeles indica extract, and its isolated constituents against H. pylori growth were examined by Rao et al. (2012). Among the tested solutions, ethanol extract and one pure constituent, ovatodiolide (OVT) showed the highest antimicrobial activity. It highlighted the bactericidal effect of the OVT on H. pylori standard strain, as well as multidrug-resistant strains. Moreover, it was demonstrated that OVT inhibited the H. pylori bacteria adhesion and invasion to human gastric epithelial (AGS) cells. These results indicate that OVT might be useful as a food supplement or drug development for H. pylori complications (Rao et al., 2012). In the same direction, other studies have also focused on the antihelicobacter action of pure constituents, based on the herbal extracts already evidence-based activity. Therefore, Sisto et  al. studied artemisone (artemisinin derivative) to examine its in vitro activity against H. pylori. Some other molecules of the same class were included in the study, together with their combination with standard antibiotics usually prescribed for H. pylori infections.

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The possible antibacterial effect was assessed against a number of 24H. pylori strains isolated from clinical cases, with different antibiotics susceptibilities. The authors found that artemisone own synergistic effects with amoxicillin in 60% of strains, with clarithromycin in 40%, and with metronidazole in 20%. They also claim that this was the first in  vitro experimental demonstration of the significant activity of artemisinin derivatives against intracellular H. pylori, thus confirming the potential of this compound for the treatment of H. pylori infection, ­especially in combination with antibiotics (Sisto et al., 2016).

3.4.3  Herbal Extracts With Activity Against Respiratory Diseases The number of plant extracts used in respiratory diseases is very high. It is important that it corresponds to the phase or form of coughing. As is known, coughing is only a symptom in which a category of plants is indicated in the initial stage of irritation, another in larynopharyngitis or incipient bronchitis and another category of plants are useful in chronic bronchitis. Smooth-action plants have as basic active ingredients the mucilagies. In popular medicine, the most widely used mucilaginous species are common marsh-mallow (Althaea officinalis), sea-buckhorn (Hippophae rhamnoides), plantain (Plantago lanceolata), and hedera (Hedera helix leaves). These plants have action to soothe the mucosal irritations and to alleviate their excitability. A. officinalis is recommended in case of lipemia, inflammation of nasal and oral cavities, gastric ulcer, platelet aggregation, cystitis, and irritating coughs. Its antioxidant activity has also been demonstrated, accounting for approximately 69% of the activity of the reference compound alpha-tocopherol (Gautam et  al., 2015; Sadighara et  al., 2012). A. officinalis is one of the most spread medicinal plants from Malvaceae family. It has been used in the traditional medicine all over the world when treating inflammatory diseases of the upper respiratory system and irritational coughing. It is also used for oral tissue inflammations and gingival abscesses (Gautam et al., 2015). P. lanceolata (plantain) as one of the major species used as a medicinal plant is commonly used in the treatment of common colds, and the associated symptoms to soothe and suppress cough, or as an antiviral, antimicrobial, or antioxidant agent (Boskabady et al., 2006; Ferrazzano et al., 2015). According to Zhou et al., an extract obtained from root of Sorghum bicolor (L.) (RSB) proved to be effective for the treatment of cough with potent antitussive, expectorant, and bronchodilating activities. S. bicolor a medicinal plant used in the traditional medicine as a remedy for cough and asthma. In the antitussive tests performed, the

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t­ reatment with the aforementioned herbal extract significantly inhibited the frequency of cough, and prolonged the cough latent period in animals. An exact dose of 200 mg/kg in mice and 100 mg/kg in guinea pigs resulted in therapeutic effect equal to codeine phosphate 20 mg/ kg (standard antitussive drug). In the expectorant evaluation, 50, 100, and 200 mg/kg of extract significantly increased the amount of phenol red output for 0.39-, 1.18-, and 1.96-folds in mice tracheas. In the bronchodilating test, the RSB treatment at 100 mg/kg extended the preconvulsive time for 44.84% compared with that of before treatment in guinea pigs (Zhou et al., 2013). A significant antitusive and expectorant effect was observed by Tao Guo et  al. with the herbal extract of Potentilla anserina. The root of P. anserina L. (Rosaceae) is an herbal medicine that has been used as an antitussive and expectorant drug for thousands of years in Chinese folk medicine. The study estimated the antitussive and expectorant effects of P. anserina extract to validate its traditional use (Guo et al., 2016). A standardized combination of aqueous ethanolic extracts is already approved in Scadinavia for the treatment of infections of respiratory tract. The herbal beverage is named as Kan Jang and is obtained by a mix of aqueous ethanolic extracts of the following plants: Echinacea purpurea (L.) root, Justicia adhatoda L. leaf, and Eleutherococcus senticosus root. A clinical 5 days trial comparing the effect of Kan Jang, placebo, and bromhexime—a standard drug usually prescribed in the upper respiratory infections, was set up by Barth et al. The study investigated two directions of research in the same time, namely, cough relief—evaluated by the cough index (change of cough frequency) and second, safety issues were observed. The safety was assessed in terms of reported adverse events and hematological data. The results support the therapeutic use of the standardized herbal extract, as Kan Jang presented good tolerability and safety profile, and the patients response to the treatment was an improvement of the cough index in the upper respiratory infections (Barth et al., 2015). Krawitz et al. (2011) have studied the antimicrobial activity of standardized extract prepared from elderberry fruits. S. nigra L. fruits are well known as agents to support the body against cold and common flu. It is also known that bacterial superinfection during a flu virus infection can lead to severe pneumonia. A standardized elderberry fruit extract (produced by Rubini, BerryPharma AG) was analyzed to establish its antimicrobial and antiviral activity against three Gram-positive bacteria and one Gram-negative responsible for the upper respiratory tract infections, and experiments on cell cultures for two different strains of influenza virus. The antimicrobial activity of the elderberry extract was determined by experiments carried out on bacterial cultures in liquid medium using the plant extract at concentrations of 5%,

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10%, 15%, and 20%. The inhibitory effects were determined on human pathogens isolated from clinical cases. The experiments carried out demonstrated for the first time that a standardized fruit extract has antimicrobial activity against Streptococcus pyogenes or Branhamella catarrhalis strains in liquid cultures.

3.4.4  Herbal Extracts for the Prevention of Dental Disorders Dental caries is considered to be a preventable disease, and various antimicrobial agents have been developed for the prevention of dental diseases; however, many bacteria show resistance to existing agents. Yim et  al. reported the antimicrobial action of 14 medicinal against five common oral bacteria as a screen for potential candidates for the development of natural antibiotics. Aqueous herbal extracts were tested against E. faecalis, Actinomyces viscosus, Streptococcus salivarius, Streptococcus m ­utans, and Streptococcus sanguis. Most of the aqueous herbal extracts demonstrated antimicrobial activity against the five types of pathogenic oral bacteria. The extracts of Sappan lignum, Coptidis rhizoma, and Psoraleae semen significantly inhibited the growth of oral bacteria (Yim et al., 2013). S. mutans and Lactobacillus acidophilus are the two main microorganisms responsible for dental caries (Cagetti et al., 2013). Cariogenic properties of S. mutans are due to its adhesion to dental surfaces, and ability of living in acidic environment. According to Haghgoo et  al. (2017), the root ethanolic extract of A. officinalis exhibited antibacterial effects on S. mutans and L. acidophilus, but this effect was less than those of chlorhexidine mouthwash and penicillin. The antibacterial effect increased with an increase in the concentration of the extract. S. mutants and L. acidophilus response to herbal extracts was also reported by Nagarajappa et  al. More exactly herbal extract prepared from Hibiscus rosa-sinensis and of Jasminum grandiflorum leaves significantly inhibited the in vitro growth of L. acidophilus and S. mutans (P ≤ 0.05). L. acidophilus and S. mutans are two cariogenic bacteria of medical importance in the dental disorders, especially identified at children. The herbal extracts tested were obtained by the hot and cold extraction methods. Both plant extracts proved good in vitro antimicrobial effects. Nevertheless, the authors concluded that, clinical trials on the effect of these plants are essential before advocating large-scale therapy (Nagarajappa et al., 2015). Ajagannanavar, S. L. evaluated the in vitro activity of aqueous and alcoholic Glycyrrhiza glabra (licorice) extract against S. mutans and L. acidophilus in comparison with chlorhexidine. The antibacterial activity of the licorice root extract against the growth of S. mutans and

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L. acidophilus was higher than that of the aqueous form and chlorhexidine. The reasons may be better solubility of the licorice compound in alcohol or the very presence of alcohol (Ajagannanavar et al., 2014). An extensive review of Shekar et al. pointed not  0.9 and women > 0.85 Urinary albumin excretion ≥20 μg/min or albumin: creatinine ≥30 mg/g

Dyslipidemia Central obesity Microalbuminuria

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Table 14.2  Diagnostic criteria of ATP III for the diagnosis of metabolic syndrome Risk Factor

Criterion

Abdominal obesity Triglycerides HDL Blood pressure Fasting blood glucose

Waist circumference in men > 102 cm and women > 88 cm >150 mg/dL or pharmacological treatment for hypertriglyceridemia Men > 40 mg/dL and women > 50 mg/dL >130/85 mmHg or pharmacological treatment for hypertension >100 mg/dL or pharmacological treatment for glucose elevation

­ xidative pathways in mitochondria, due to a high intake of simple caro bohydrates in a chronic manner, leading to an increase in free oxygen, responsible for the nonenzymatic production of ROS and decrease in the production of ATP, given the oxidative alterations generated in the mitochondria, this state of toxicity is termed as mitochondrial dysfunction. The collapse of the mitochondria due to the establishment of oxidative stress causes disturbances in the level of subcellular organelles and cellular lesions that ends with the rupture of the plasma membrane and cell necrosis (Hernández et al., 2013). The necrosis comprises an irreversible state of the cell, where the integrity of the plasma membrane cannot be maintained and there is an escape of cytoplasmic elements, denaturation of the proteins by autolysis or coming from lytic enzymes of neighboring leukocytes, since the necrosis attracts the components of inflammation. The severity of the above is exacerbated when the damage is in cells that do not have regenerative capacity, such as neurons, nephrons, and retinocytes, associated with the microvascular complications of DM: neuropathy, nephropathy, and retinopathy, respectively, that within the framework of MS patients usually show signs of alterations and dysfunction in the neurological and renal system. On the other hand, the excessive circulation of free fatty acids, mainly saturated ones, as part of the lipotoxicity contributes to the metabolic activation of cells of the immune system mediated by tolllike receptors (TLRs) which induces the activation of the nuclear factor Kappa beta (NF-kB) and with it the synthesis of proinflammatory cytokines such as tumor necrosis factor alpha (TNF-α), interleukin-1 (IL-1), interleukin-6 (IL-6), and the chemoattractant protein of ­macrophages-1 (MCP)-1) as well as the reduction or inhibition of the expression of anti-inflammatory cytokines (Hirai et  al., 2010), generating a low-grade pro-inflammatory state. The chronic inflammatory state, generated by obesity and glycolipotoxicity, is the main

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triggers of IR. Inflammatory cytokines and lipotoxicity bring about the activation of intracellular kinases such as C-Jun N-Terminal kinase (JNK) and nuclear factor-κB (IκB) kinase (IKK) β, activating enzymes of the inflammatory pathways and catalysts of phosphorylation of the insulin receptor substrate (IRS-1) in specific serine residues. The phosphorylation of IRS-1 inhibits the signal transduction mediated by this substrate and catalyzes its proteosomal degradation, causing partial inhibition of insulin signaling (Nieto-Vazquez et al., 2008). On the other hand, the increased circulation of free fatty acids increases the synthesis rate of triacylglycerides in the liver, thus increasing the concentration of plasma lipoproteins and contributing to the dyslipidemia characteristic of MS (Ebbert and Jensen, 2013). Likewise, the increase in plasma concentrations of proinflammatory cytokines activates the vascular endothelium (VE), which will express adhesion molecules (ICAM/VACM) and the alteration of its metabolism in its apical membranes. The activation of the VE induces the adhesion of leukocytes and with it induces the ROS generation; this act favors the endothelial dysfunction, the atherosclerosis, and the increase in the arterial pressure. In the VE, cell signaling mediated by IRS-1 is necessary for the production of nitric oxide synthase (NOS) stimulated by insulin. Therefore, the phosphorylation of IRS-1 brings about the inhibition of synthesis of NOS, the main enzyme producing nitric oxide (NO), regulator of vasodilation. In turn, other vasoconstrictor molecules such as endothelin-1 (ET-1) and angiotensin-II maintain the VE stimulation, promoting alterations in the regulation of vascular diameter and thus blood pressure (Arce-Esquivel et al., 2013). Inflammatory cytokines regulate a large number of cell signaling processes. Part of the effect of these cytokines is the increase in the hepatic synthesis of acute phase proteins and coagulation factors. Other tissues and systems regulate other signaling pathways involved in proteolysis, at the muscle level, appetite regulation and modification of nervous tone in the central nervous system; increased secretion of ­catecholamines and cortisol, in the endocrine system; and modification in the leukocyte profile, at hematological and immune level (Gruys et al., 2005; Lumeng and Saltiel, 2011). The pathophysiology of MS is really complex since a large number of cell signaling pathways are altered, in the response to insulin being essential for clinical manifestations. However, the chronic inflammatory state and oxidative stress play a major role in the degenerative process and in the exacerbation of the signs and symptoms of MS. Currently, the role of various environmental factors in the development of the disease as well as nutrition is recognized, such as the level of physical activity, drug use, and access to health services, among others. One of the environmental factors that has become

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more ­important in recent years is the microbiome, whose dysbiosis has been widely related to the development of diseases that present a degree of chronic inflammation (Slingerland et al., 2017).

14.3  Role of the Microbiota in the Development of the MS The term microbiota refers to the community of microorganisms living in a certain ecological niche. Among the main functions of the microbiota are: (a) supply of essential nutrients; (b) development and modulation of the immune system; and (c) microbial antagonism (Young, 2017). The microorganisms that make up the microbiota are determined by the types of nutritional sources, the profiles of omnivores, carnivores, and herbivores being different. The characteristics of the diet, together with the genetic factors, influence the predominance of some microorganisms over others. The initial microbiota generates metabolites that can have a beneficial effect on the carrier such as anti-inflammatory and antioxidant actions, regulation of the intestine barrier function, as well as the production of vitamins and energy sources (HMPC, 2012). The microbial ecosystem of the intestine (intestinal microbiota) includes many native species that permanently colonize the gastrointestinal tract, and a variable series of microorganisms that only do so transiently. To the set formed by the microorganisms, their genes and their metabolites are called microbiome. The human being has 100 trillion microorganisms in the intestine, a figure that is calculated to be 10 times higher than the number of cells in the human body, so the commensal bacteria and the fungi that inhabit the body greatly outnumber the cells in human beings (Icaza, 2013). The number and variety of bacteria increase exponentially from the proximal end of the gastrointestinal tract to the distal end, with the colon harboring most of the intestinal microbiota. The anatomical, histological, chemical, and biochemical conditions of the digestive system are a determining factor for the development of microorganisms, thus generating specific niches in different portions of said apparatus (Icaza, 2013). Intestinal colonization is a dynamic process influenced by factors such as the nutritional status and microbiota of the mother, the gestational age of the newborn, the type of delivery, breastfeeding and its microbiological quality, the complementary feeding of the infant, and the use of antibiotic therapy in the mother or child (Browne et al., 2017). The colonization begins at birth when the newborn, practically sterile, meets its environment and the first microbial biofilms.

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The newborn acquires bacteria such as bifidobacteria and lactobacilli from the birth canal. Children born by caesarean section are initially colonized by bacteria from the hospital environment and the skin, exhibiting a lower pattern of protective bacteria (Sánchez et al., 2017). The intestinal microbiota of newborns fed only breast milk has a predominance of bifidobacteria, while children who receive artificial lactation have a more complex and diverse microbiota, with members of the families Enterobacteriaceae and Enterococcus. It is speculated that this differential colonization has a protective effect against the inflammatory microenvironment induced by the immunogens transferred through artificial lactation, important in sensitivity or resistance to the development of chronic diseases of inflammatory origin (Gritz and Bhandari, 2015). Microbiota plays an important role in maintaining the homeostasis of the individual. Competes for nutrients, receptors, and displaces pathogens, produces antimicrobial factors, regulates the turnover rate of enterocytes, promotes the development and differentiation of epithelial cells, fortifies the intestinal barrier, and maintains the proper functioning of mucosal immune system by inducing IgA secretion (Gritz and Bhandari, 2015). The physical, chemical, and immune union of the intestinal barrier are pillars in the maintenance of the number and location of the microbial population and of the beneficial effects on health that this entails. The intestinal epithelial cells are constantly subjected to cytotoxic, metabolic, and pathogenic stress, situations that can produce a break in the intestinal barrier causing the passage of microbial components into the bloodstream resulting in a pro-inflammatory response (Sommer et al., 2017). The dysbiosis (changes in the intestinal microbiota and the adverse host response) has been associated with important conditions for the development of the characteristic alterations of the MS. In people with MS an increase in the Firmicutes/Bacteroidetes ratio is exhibited. The Bifidobacteria and the Bacteroides spp. seem to be protective against the development of chronic inflammation characteristic of the MS. This could have a microbial component, with probable therapeutic implications (Sommer et al., 2017). The MS has been associated with an increase in the relative abundance of Firmicutes and proportional reductions in Bacteroidetes, by comparing the composition of the intestinal microbiota of genetically obese (leptin-deficient mice ob/ob) and skinny mice (Ley et al., 2005). This was confirmed by Waldram et  al. (2009) who stated that there is also a decrease in Bifidobacterium and an increase in Halomonas and Sphingomonas in the intestinal microbiota of genetically obese Zucker rats (fa/fa), in comparison with control rats. Changes in the relative proportions between Firmicutes and Bacteroidetes of the gut

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­ icrobiota have also been associated with obesity in humans. In m addition, obese humans, following a hypocaloric diet (low in carbohydrates or fats), showed significant increases in the proportions of Bacteroidetes parallel to weight loss during a 1-year intervention period (Ley et al., 2006). Mice genetically modified for the development of obesity (ob/ob), present 50% less Bacteroidetes and more Firmicutes than their slender brothers. On the other hand, the colonization of mice with the microbiota of normal mice produces a decrease in fat in 10–14 days, independent of a variation of the intake of simple carbohydrates and saturated fatty acids (Ley et al., 2005). By supplying normal-weight mice with a typical western diet high in simple carbohydrates for 8 weeks, a marked reduction in Bacteroidetes and a marked increase in Firmicutes were observed. Jumpertz et  al. (2011) administered 12 thin people and 9 people with obesity, variable diets in caloric content and compared the calories ingested with the fecal calories. The modification of the secondary microbiota to the diet showed a 20% increase in Firmicutes and corresponding decrease in Bacteroidetes, accompanied by an increase in energy recovery of approximately 150 kcal. Findings such as those described above have led to the hypothesis that the microbiota of people with obesity may be more efficient in extracting energy than the microbiota of thin individuals. By virtue of the above, situations that occur around birth and modify the composition of the intestinal microbiota, increase the risk of developing obesity, diabetes, and CVD in adulthood (Martínez et al., 2017). The intestinal microbiota, although beneficial, must be maintained within specific limits. It has been argued that modifications in the intestinal microbiota lead to a chronic state of endotoxemia having as a consequence a process of chronic inflammation, which is a key factor associated with the increase in adiposity. In Table  14.3, some mechanisms are described where dysbiosis is related to the development of MS. The main association between the development of a proinflammatory response and the intestinal microbiota lies in the recognition of bacterial recognition receptors. Currently, two types of receptors have been studied, the so-called TLRs and NOD-type receptors (NLRs). Most TLRs are surface receptors whereas NODs (NOD1 and NOD2) are cytoplasmic (Yiu et al., 2017). Cell recognition receptors of cells of the innate immune system, such as TLR receptors, constitute a starting point for immunity, which is activated in response to microbial- or dietary-derived stimuli (proteins or lipids) and informs cells of the immune system so that they respond appropriately to these. After activation by a ligand, the TLRs interact with different proteins that activate the transcription of different factors (such as MAPKs and NF-kB), and

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Table 14.3  Mechanisms of dysbiosis related to the development of metabolic syndrome Mechanism

Description

Bacterial fermentation

Alterations in the bacterial fermentation of the carbohydrates of the diet, which cannot be digested by the host, with the consequent decrease in the production of monosaccharides and short-chain fatty acids (SCFA). The SCFA are substrates of the colonocytes that bind to specific receptors of endocrine intestinal cells (GRP43 and GRP41) which increases the YY peptide, leading to the delay of intestinal transit, increasing the absorption of nutrients and leptin levels. The decrease in SCFA in people with metabolic syndrome is related to a lower satiety response The dysbiosis in metabolic syndrome is associated with greater gene expression that promotes lipogenesis and fat deposition in adipocytes. The decrease in intestinal expression of the adipose factor induced by fasting (FIAF) favors the capture of fatty acids and the expansion of adipose tissue The microbiota increases the vascularization induced by inflammation and mucosal blood flow, which increases the absorption of nutrients. The intestinal microbiota is able to promote a state of low-grade systemic inflammation, insulin resistance, and increase cardiovascular risk through mechanisms that include exposure to bacterial products, in particular, LPS derived from Gram negative bacteria. This is called metabolic endotoxemia

Microbial regulation

Increase of vascularization

the synthesis of different cytokines and immunological mediators of inflammation (Caricilli et al., 2011). In this regard, it has been shown that lipopolysaccharide (LPS) of the cell membrane of Gram-negative bacteria, such as Firmicutes, can act as a receptor ligand TLR4 and TRL2, which have the function of stimulating the release of endogenous inflammatory cytokines such as TNF-α, IL-6, and other pro-inflammatory cytokines related to the induction of IR. Therefore, the increase in Firmicutes implies an important factor in the development of inflammation (Ghoshal et al., 2017). Histochemical studies show that in people with MS there is an increase in LPS in the colonocytes. The LPS from the death of Gramnegative bacteria is transported to the bloodstream by chylomicrons (synthesized in greater quantities as a result of a high-fat diet). In the bloodstream, LPS is recognized by the CD14/TLR4 receptor of macrophages, activating a signaling pathway that ends with the activation of NF-κB and subsequent synthesis and release of cytokines TNF-α, IL-1, and IL-6, leading to a state of chronic inflammation, ABCD, and IR. The blood LPS increases when there is a rupture in the integrity of

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the intestinal barrier, a high permeability and less degradation of LPS by intestinal alkaline phosphatase (Ghoshal et al., 2017; Ubeda et al., 2012). Based on the foregoing, a restoration of Firmicutes/Bacteroidetes relationship, through the administration of probiotics, is suggested as an alternative in the treatment of diseases of inflammatory origin. The use of probiotics, as food or nutraceutical, is a viable therapeutic option, which should be considered within the framework of the diet therapy treatment of people with MS.

14.4  Characteristics of Probiotics The WHO defines probiotics as “live microorganisms that, when administered in adequate amounts, confer a benefit to the consumer’s health” (FAO, 2001). In principle, any component of the occupation microbiota could be a candidate to become a probiotic, since all of them potentially participate in the benefits granted by the group. However, in practice they mainly belong to two microbial groups: the lactobacilli and the bifidobacteria. The reason for this is that they are probably the only ones, among those that colonize the mucous membranes, that are innocuous under almost any circumstance and that, therefore, have been recognized as GRAS (generally regarded as safe) organisms and QPS (qualified presumption of safety) by the Food and Drug Administration of the United States and the European Food Safety Authority, respectively (Szajewska et al., 2014). Among the desirable characteristics of a probiotic are: (1) adaptation to the conditions of the target cavity and a good adherence to the epithelium that covers it (that is why organisms with the same origin are preferred), (2) generation of antimicrobial substances; (3) absence of resistance transmissible to antibiotics; and (4) existence of clinical trials that certify the beneficial effect on health. Probiotics include a large number of genera of microorganisms, such as Lactobacillus, Bifidobacterium, Saccharomyces, Streptococcus, and Enterococcus. In turn, the genus Lactobacillus comprises >90 species, the most commonly used include Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus bulgaricus, Lactobacillus plantarum and Lactobacillus reuteri. Most of the clinical trials reported to date have used mixtures of different probiotics mainly Lactobacillus spp. in combination with another probiotic (Hill et al., 2014). The dose of probiotics needed for a biological effect varies enormously depending on the strain and the product. Although many over-the-counter products provide between 1 and 10 billion CFU/ dose, some products have been shown to be effective at lower ­levels, while others require larger amounts, so it is not possible to ­establish

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a general dose for probiotics. The minimum criteria required for ­probiotic products are that the probiotic must: (1) be specified by ­gender and strain, research on specific probiotic strains cannot be applied to any product marketed as probiotics; (2) contain live bacteria; (3) be administered in adequate dose until the end of its useful life (with minimum variability from one batch to another); and (4) have been shown to be effective in controlled studies in humans (ILSI, 1999). Since standards for content and label declarations on products are not universally established and/or not universally applied, industry must maintain integrity in formulation and labeling so that consumers can rely on this category of products.

14.5  Mechanisms of Action of Probiotics The intestinal microbiota is responsible for producing metabolites that function as signaling molecules at the systemic level, affecting the host’s metabolism. These metabolites directly affect the function of different organs, including the intestine, liver, brain, adipose tissue, and muscle (Tremaroli, 2012). Likewise, the microbiota contributes to the enzymatic digestion in the digestive tract, through the production of enzymes capable of degrading polysaccharides and bile acids (Tremaroli, 2012), and capable of modifying the phytochemicals consumed in the diet (Laparra and Sanz, 2010). Thus, the mechanisms by which they offer a health benefit are several, and will be discussed more thoroughly in this section.

14.5.1 Fermentation of Polysaccharides Foods of vegetable origin provide nondigestible carbohydrates for human enzymes. However, these carbohydrates are susceptible to enzymatic degradation and fermentation by the intestinal microbiota. Some species of microorganisms such as bacteroides ovatus have double the enzymes glucosidases and liases encoded in their genome, compared to humans (Tremaroli, 2012). The nondigestible poly-oligosaccharides are considered prebiotics, which have a favorable effect on the growth of the commensal intestinal microbiota. Likewise, some microorganisms such as lactobacilli and bifidobacteria are able to ferment the said prebiotics and produce short-chain fatty acids (SCFAs) (Laparra and Sanz, 2010). The SCFAs are composed of acetic, butyric, and propionic acids. These compounds have a wide variety of biological effects, such as the regulation of immune function, liver metabolism, and as an energy substrate of intestinal epithelial cells (Laparra and Sanz, 2010). The GPR41/FFAR3 and GPR43/FFAR2 receptors are two G-protein-coupled receptors that have affinity for SCFA, found in

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e­ nteroendocrine, immunological, and adipocyte cells. At the intestinal level, SCFA binds to the GPR41 receptor in enteroendocrine cells, stimulating the secretion of peptide YY, an intestinal hormone that induces satiety and reduces food consumption (Samuel et  al., 2008; Cani et al., 2009). On the other hand, the binding of SCFA to intestinal receptors GPR43 promotes the secretion of incretins, increasing the sensitivity and secretion of the host’s insulin (Tolhurst et al., 2012; Ezcurra et al., 2013). In adipose tissue, adipocytes express both receptors, GPR41 and GPR43. The stimulation of the first by SCFA stimulates the synthesis and secretion of leptin, generating a feeling of satiety mediated by the arquato nucleus of the hypothalamus. On the other hand, the binding of SCFA with GPR43 suppresses insulin signaling, reducing adipogenesis and lipogenesis (Li et al., 2017). Also, butyrate is a potent inhibitor of the enzymes histone acyltransferase and deacetylase, thus regulating epigenetically the proliferation and differentiation of the immune system (Chang et al., 2013), and the biogenesis of the mitochondria of different cells and increasing their capacity for beta oxidation. In turn, it directly stimulates pancreatic beta cells to increase insulin secretion (Li et al., 2017). On the other hand, SCFAs have a regulatory effect on the metabolism of fatty acids. Through the activation of AMP-activated protein kinase (AMPK) in liver and muscle tissue cells (Hu et al., 2010), SCFA increases the expression of the peroxisome proliferator-activated receptor gamma coactivator (PGC)-1α, which controls the transcriptional activity of diverse transcription factors, including the peroxisome ­proliferator-activated receptor (PPAR) α, PPARγ, and PPARδ, among the main ones, which regulate the metabolism of cholesterol, glucose, and lipids. As a result of the regulation of transcription factors, it is possible to reduce the de novo fatty acid synthesis in the liver, and increase the oxidation of fatty acids at muscular and hepatic levels, as well as reduce the synthesis of cholesterol (Den Besten et al., 2013; Hu et al., 2010). Likewise, the synthesis of SCFA by probiotic strains reduces the colon pH, which favors the growth of beneficial bacteria, among them are lactobacillus and bifidobacterium. In turn, the latter increases the production of SCFA, favors a healthy immune response, and reduces the growth of pathogenic microorganisms, thus reducing inflammation and the frequency of gastrointestinal disorders (McLoughlin et al., 2017). The consumption of probiotics with microorganisms that favor the fermentation of nondigestible carbohydrates or prebiotics, promotes the generation of SCFA which in turn provides a variety of biological effects. Likewise, it is important to highlight the need to consume an adequate amount of fiber in order to encourage the synthesis of SCFA, and not the consumption of probiotics alone. Thus, it is possible to

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f­avor the metabolic control of the patient with MS by means of the wide variety of biological effects exerted by the SCFA (McLoughlin et al., 2017).

14.5.2 Regulation of Bile Acid Metabolism As mentioned, the composition of the gut microbiota is altered by different factors, with bile acids being one of the main regulators. Studies on patients with liver cirrhosis, whose bile acid production has reduced with the advancement of the disease, have shown bacterial dysbiosis and a significant reduction in biological acids in feces (Ridlon et al., 2014). On the other hand, the intestinal microbiota plays a bilateral role in the synthesis and degradation of bile fatty acids. Through the expression of hydrolases of bile salts, intestinal microorganisms promote deconjugation of bile acids, thus preventing their enterohepatic resorption. The reduction in bile acids, due to its degradation, stimulates feedback mechanisms at the hepatic and intestinal level, increasing the expression of the nuclear receptor farnesoid x receptor (FXR). This receptor regulates the expression of the hepatic enzyme 7α-hydroxylase (CYP7A1), which is the limiting enzyme in the synthesis of bile acids (Ridlon et al., 2014). As a result, the synthesis of bile acids increases, while maintaining a reduction in the pool of bile acids from the gallbladder, which increases the excretion of cholesterol by fecal route and reduces the risk of vesicular lithiasis. Likewise, the degradation and conversion of bile acids by bacteria in the microbiota reduces the absorption of lipids and intestinal cholesterol. However, Bacteroides intestinalis, Bacteroides fragilis, and Escherichia coli are potent generators of enzymes that convert bile acids into possible carcinogens. Therefore, the restoration of a healthy microbiota, through the use of probiotics, can reduce the absorption of lipids and the production of metabolites with carcinogenic activities (Laparra and Sanz, 2010).

14.5.3 Choline Microbial Metabolism Choline is a component of various biomolecules, but fundamental in the phospholipids of cell membranes and play an important role as a methyl-group donor in methionine metabolism, it also participates in the synthesis of the neurotransmitter acetylcholine (Michel et  al., 2006). This nutrient is obtained mainly from dietary sources, since in spite of existing endogenous production, this appears to be insufficient in some cases. Also, choline is essential for lipid metabolism and the synthesis of very low-density lipoproteins in the liver, so a consumption, absorption, or poor production is associated with

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alterations of the intestinal microbiota and development of hepatic steatosis (Tremaroli, 2012). The alteration of the gut microbiota in obesity and/or MS may be accompanied by an increase in the microorganism involved in the transformation of choline to toxic metabolites such as trimethylamine and reduce nutrient absorption. The decrease in the bioavailability of choline suggests a triggering factor for nonalcoholic fatty liver, which is also related to IR and alteration in the blood lipid profile (Tremaroli, 2012).

14.5.4 Regulation of Permeability and Inflammation The intestinal permeability refers to the capacity of this organ to allow the passage of substances from the epithelium or mucosal layer to the submucosa or enter the systemic circulation. In recent years, it has been discovered that obesity and other diseases with a background of chronic inflammation have an alteration (increase) in intestinal permeability. Thus, allowing the passage of unwanted substances, such as LPS, which trigger an inflammatory response that can become systemic. Likewise, some enteropathogenic microorganisms obtain access to the body through the alteration in intestinal permeability (Bischoff et al., 2014; Boulangé et al., 2016). The intestinal permeability depends on the tight junctions, which seal the paracellular space, regulating the passage of water, ions, and small molecules. The adherence junctions are another type of junction that is found in this space, and these in turn are important in cell– cell signaling processes (Bischoff et al., 2014). Some probiotics, such as the bacterium E. coli Nissle 1917, have been shown to reduce alterations in intestinal permeability due to pathogenic microorganism species. Said probiotic increases the expression of ZO−2 proteins, which play an important role in tight junction. On the other hand, said probiotic also increases the expression of other proteins related to tight junction, such as claudin−14. Therefore, the increase in the expression of said proteins reduces intestinal permeability (Bischoff et al., 2014). The reduction in intestinal permeability is an important protection mechanism to prevent translocation of bacteria or bacterial products (e.g., LPS) to the submucosa or circulation. Thus, contributing to the reduction in the activation of the immune system, mediated by TLR, and reduction in the consequent inflammatory response (Bischoff et al., 2014).

14.5.5 Metabolism of Dietary Phytochemicals Many of the dietary phytochemicals, which offer biological activities beneficial to health, are in the form of glycoconjugates, polymers, or esters, which are not bioavailable in that form. Most polyphenols, a class

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of phytochemicals, are bound to highly hydrophilic polar molecules, which does not allow their absorption by passive diffusion into the small intestine enterocytes (Laparra and Sanz, 2010; Rossi et al., 2013). Many reactions that transform phytochemicals to bioactive molecules require enzymes produced by colonic microbiota. Probiotics such as lactobacilli and bifidobacteria can affect the bioavailability, biological activity, and kinetics of the phytochemicals consumed (Rossi et al., 2013). So, knowing the main ways in which the microbiota and/ or probiotics can alter the phytochemicals play an important role in the context of functional nutrition. Some bacterial strains, including bifidobacteria, synthesize a variety of glycosyl hydrolases, mainly β-glucosidase, which are necessary to process the nondigestible polysaccharides that reach the large intestine. Among the bifidobacteria that produce these enzymes Bifidobacterium adolescents, Bifidobacterium animalis, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium longum subsp. infantis, and Bifidobacterium pseudocatenulatum are the main ones. Thus, these enzymes are important part of the digestion of food and the processing of polyphenols that improve their bioavailability (Rossi et al., 2013). In addition, lactobacillus species that produce β-glucosidase enzymes have also been identified. Among the main species are L. acidophilus, L. casei, Lactobacillus paracasei, L. rhamnosus, and L. plantarum. However, lactobacilli have been less studied than bifidobacteria because of their ability to hydrolyze glucoconjugates from food (Rossi et al., 2013).

14.5.6 Antioxidant Properties Several studies with different probiotic bacterial strains have found that they exert some antioxidant capacity. Although little is known about the mechanism of bacterial action to modulate the formation of free radicals, systematic reviews have shown that the use of probiotics improves the oxidative status of the host (Mishra et al., 2015; Wang et al., 2017). Several mechanisms have been proposed, including the ability to chelate metal ions. Some ions (Cu+2 and Fe+2) catalyze the conversion of H2O2, a reactive oxygen species, to a more reactive and oxidizing form. The study of Streptococcus thermophilus 82 and L. casei KCTC 3260 showed that both bacterial strains have antioxidant effects on the host, by chelation of Cu+2 and Fe+2 (Wang et al., 2017). On the other hand, probiotics, like humans, have an antioxidant enzyme system. Among the main antioxidant enzymes produced by probiotics are the superoxide dismutase and catalase. Bacteria express the said enzymes to adapt to media where there is an increased amount of ROS. Studies that have evaluated the usefulness of ­probiotics that

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express these enzymes have found beneficial effects in pathological conditions where there is an increase in ROS (De LeBlanc et al., 2008; Wang et al., 2017).

14.5.7 Antimicrobial and Immunomodulatory Properties Chronic inflammation is a characteristic of MS and has a bilateral interaction with the intestinal microbiota, that is, the pro-inflammatory environment promotes a remodeling of the microorganisms, at the same time that said remodeling induces greater inflammation. Some bacterial strains have been shown to have anti-inflammatory effects, including L. reuteri. Through the production of different inhibitory factors of the immune activation mediated by LPS, the said lactobacillus reduces the secretion of TNF-α (Jones and Versalovic, 2009). On the other hand, L. reuteri is also recognized for expressing peptides with antimicrobial properties, including reuterin (Langa et al., 2013). The said peptides reduce the growth of pathogenic bacterial strains, reducing the inflammation and the condolence time of gastrointestinal diseases. The antiinflammatory activity of probiotics is specific to each strain. Among the probiotics with antiinflammatory properties are L. rhamnosus GG, Propionibacterium freudenreichii ssp. shermanii JS, and Bifidobacterium animalis ssp. lactis Bb12. A study conducted by Kekkonen et al. (2008) showed that L. rhamnosus and P. freudenreichii are able to reduce the levels of highly sensitive C-reactive protein in healthy people, while B. animalis reduced the plasma levels of IL-2. The mechanisms by which these strains reduce the markers of inflammation are not yet well understood. Although it has been recognized that a significant variety of probiotics possess anti-inflammatory activities, not much is known about their mechanisms of action. An important part has been attributed to modulate the activation of TLRs (Plantinga et  al., 2011). Also, some studies have found that the interaction of probiotic strains with dendritic cells in the intestine induces the production of IL-10, which has anti-inflammatory and immunomodulatory activities (Kang and Im, 2015). On the other hand, some probiotics induce an increase in the synthesis of immunoglobulins M, A, and G, and induce a higher production of IL-12, favoring a Th1-type response (mediated by cells) and reducing a Th2 response (Ng et al., 2009).

14.6  Probiotic Beverage as a Therapeutic Option in MS Probiotics can be consumed through a food or in nutraceuticals, in a balanced and adequate way. There is a wide variety of probiotic

Chapter 14  Probiotics Beverages: An Alternative Treatment for Metabolic Syndrome   475

dairy products (Kanmani et al., 2013). In the present review, probiotic beverages and their benefits in the MS have been pointed out, specifically in the reduction or improvement of some biochemical markers. Owing to their nutritional contribution, and sensory and functional benefits in food, during the last decades, the use of probiotic microorganisms, such as L. acidophilus, L. paracasei among others, has increased in several types of food, especially in fermented milk drinks (Lee et al., 2015). Due to the above, clinical research has been expanded by evaluating the effectiveness of probiotic beverages on MS markers (Table 14.4). In people with moderate hypercholesterolemia [total cholesterol (TC) 5.17–7.76 mmol/L], a decrease in blood TC levels was obtained after consuming 300 g of yogurt enriched with 106 CFU of L. acidophilus and B. lactis, for 6 weeks (Asal, 2009). In turn, a double-blind placebo-controlled study conducted in Portugal on healthy women with BMI  25, LDL-c > 100 mg/dL

Group of Japanese with visceral adiposity

Dose

Results

Reference

150 mL (10  cfu/ mL) for 3 weeks 300 g yogurt daily for 6 weeks

Improvement of the total concentration of antioxidants in the blood Cholesterol-lowering effect in patients with hypercholesterolemia

Songisepp et al. (2005) Asal (2009)

125 mL of milk fermented 3 times a day for 4 weeks 106 to 108 CFU/ day for 6 weeks 200 g of milk for 12 weeks with a content of 107 CFU 1.25 × 107 CFU/g, 80 mL for 90 days. 3.7 × 106 CFU/mg 300 g for 8 weeks

LDL and HDL cholesterol levels were reduced significantly in those individuals who had cholesterol levels above 190 mg/dL Reduction of basal glucose (fasting) and glycosylated hemoglobin. Reduction of visceral adiposity.

Andrade and Borges (2009)

9

Significant reduction of cholesterol, glucose, homocysteine and IL-6. Decreased LDL levels and a significant increase in HDL levels

Ejtahed et al. (2012) Yukio et al. (2013)

Barreto et al. (2014) Mohamadshahi et al. (2014)

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in the superoxide dismutase (SOD) (antioxidant) was observed, without significant changes in the concentration of insulin (Ejtahed et al., 2012). Concluding that probiotic yogurt is a promising agent for the treatment of DM. Previously, the antioxidant power of probiotic beverages had already been demonstrated in the study conducted by Songisepp et al. (2005), which consisted of the administration of milk fermented with L. fermentum ME-3 to a group of healthy volunteers with an age range of 35–60 years, giving them the 150 mL portion daily for 3 weeks. At the end of the study, the total number of antioxidants in the blood has improved. In people with obesity, the consumption of 200 g of fermented milk with a probiotic content of 107 CFU of Lactobacillus gasseri SBT2055 (LG2055) for 12 weeks showed a reduction in visceral adiposity, with significant decreases in the abdominal perimeter (Yukio et al., 2013). Likewise, Bordalo et  al. (2015) reported a decrease in glycemic levels, TC, and triglycerides in people with MS who consumed milk fermented with L. bulgaricus and S. thermophilus. Jones et al. (2017) conducted a study to evaluate the effectiveness of microencapsulated L. reuteri NCIMB 30242 (bile salt hydrolase-active probiotic) to reduce the blood cholesterol in a formulation with yogurt, in 114 hypercholesterolemic subjects. The results were positive, after a period of 6 weeks of treatment, a significant reduction in LDL cholesterol (8.92%), TC (4.81%), and non-HDL cholesterol (6.01%) was found, compared to placebo (regular yogurt). There were no changes in serum concentrations of triglycerides or HDL cholesterol. Although clinical studies show the potential use of probiotic beverages as an adjuvant to the treatment of MS, more clinical information is still needed on the role of the microbiota and probiotics in this inflammatory disease. The beneficial effects of specific bacteria on the characteristics of MS have been described. However, given that the intestinal microbiota represents a modifiable characteristic in the intervention for the improvement of the metabolic profiles of the MS, the study of probiotics is an important tool that should be implemented in the dietotherapeutic intervention (Martínez et al., 2017).

14.7 Conclusion The microbiota plays an important role in the development and chronicity of inflammation in people with MS, which represents a modifiable feature in the intervention for the improvement of the metabolic profile in MS. Although more information is still needed to elucidate the role of probiotics as part of the treatment of MS, the findings with which they are counted, highlighted the clinical usefulness of probiotics as a strategy of nutritional intervention in the inflammatory and metabolic alterations of MS.

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References Andrade, S., Borges, N., 2009. Effect of fermented milk containing Lactobacillus acidophillus and Bifidobacterium longum on plasma lipids of women with normal or moderately elevated cholesterol. J. Dairy Res. 469–474. https://doi.org/10.1017/ S0022029909990173. Arturo A. Arce-Esquivel, Aaron K. Bunker, Catherine R. Mikus and M. Harold Laughlin (2013). Insulin Resistance and Endothelial Dysfunction: Macro and Microangio­ pathy, Type 2 Diabetes, prof. Kazuko Masuo (Ed.), InTech, DOI:https://doi. org/10.5772/56475. Available from https://www.intechopen.com/books/type-2-­ diabetes/insulin-resistance-and-endothelial-dysfunction-macro-and-microangiopathy. Asal, B.H., 2009. Cholesterol-lowering effect of probiotic yogurt in comparison with ordinary yogurt in mildly to moderately Hypercholesterolemic subjects. Nutr. Food Technol. 22–27. https://doi.org/10.1159/000203284. Barreto, F.B., et al., 2014. Beneficial effects of lactobacillus plantarum on glycemia and homocysteine levels in postmenopausal women. Nutr. J. 30 (7–8), 939–942. https:// doi.org/10.1016/j.nut.2013.12.004. Bennett, B.J., Hall, K.D., Hu, F.B., McCartney, A.L., Roberto, C., 2015. Nutrition and the science of disease prevention: a systems approach to support metabolic health. Ann. N. Y. Acad. Sci. 1352, 1–12. https://doi.org/10.1111/nyas.12945. Bischoff, S.C., Barbara, G., Buurman, W., Ockhuizen, T., Schulzke, J.-D., Serino, M., Wells, J.M., 2014. Intestinal permeability—a new target for disease prevention and therapy. BMC Gastroenterol. 14, 189. https://doi.org/10.1186/s12876-014-0189-7. Bordalo, T.L., Olbrich, M., Licursi, L., Rocha, S.M., Duarte, H.S., 2015. Clinical application of probiotics in diabetes. Crit. Rev. Food Sci. Nutr. 4–64. https://doi. org/10.1016/j.clnu.2015.11.011. Boulangé, C.L., Neves, A.L., Chilloux, J., Nicholson, J.K., Dumas, M.-E., 2016. Impact of the gut microbiota on inflammation, obesity, and metabolic disease. Genome Med. 8, 42. https://doi.org/10.1186/s13073-016-0303-2. Browne, P., Claassen, E., Cabana, M., 2017. Microbiota: In Health and Disease: From Pregnancy to Childhood. Wageningen Academic Publishers, ISBN: 978-90-8686-294-8. Cani, P.D., Lecourt, E., Dewulf, E.M., Sohet, F.M., Pachikian, B.D., Naslain, D., Delzenne, N.M., 2009. Gut microbiota fermentation of prebiotics increases satietogenic and incretin gut peptide production with consequences for appetite sensation and glucose response after a meal. Am. J. Clin. Nutr. 90 (5), 1236–1243. https://doi.org/10.3945/ajcn.2009.28095. Caricilli, A., Picardi, P., Abreu, L., Ueno, M., Prada, P., Ropelle, E., et al., 2011. Gut microbiota is a key modulator of insulin resistance in TLR 2 knockout mice. PLoS Biol. 9 (12), https://doi.org/10.1371/journal.pbio.1001212. Chang, P.V., Hao, L., Offermanns, S., Medzhitov, R., 2013. The microbial metabolite butyrate regulates intestinal macrophage function via histone deacetylase inhibition. Proc. Natl. Acad. Sci. U. S. A. 111 (6), 2247–2252. https://doi.org/10.1073/ pnas.1322269111. Daskalopoulou, S.S., Mikhailidis, D.P., Elisaf, M., 2004. Prevention and treatment of the metabolic syndrome. Angiology 55 (6), 589–612. https://doi.org/10.1177/00033197 040550i601. De LeBlanc, A.M., LeBlanc, J.G., Perdigon, G., Miyoshi, A., Langella, P., Azevedo, V., Sesma, F., 2008. Oral administration of a catalase-producing Lactococcus lactis can prevent a chemically induced colon cancer in mice. J. Med. Microbiol. 57, 100–105. https://doi.org/10.1099/jmm.0.47403-0. De Lorenzo, D., 2012. Present and future perspectives of nutrigenomics and nutrigenetics in preventive medicine. Nutrición Clínica y Dietética Hospitalaria 32 (2), 92–105.

Chapter 14  Probiotics Beverages: An Alternative Treatment for Metabolic Syndrome   479

Den Besten, G., van Eunen, K., Groen, A.K., Venema, K., Reijngoud, D.-J., Bakker, B.M., 2013. The role of short-chain fatty acids in the interplay between diet, gut microbiota, and host energy metabolism. J. Lipid Res. 54 (9), 2325–2340. https://doi. org/10.1194/jlr.R036012. Ebbert, J.O., Jensen, M.D., 2013. Fat depots, free fatty acids, and dyslipidemia. Nutrients 5 (2), 498–508. https://doi.org/10.3390/nu5020498. Ejtahed, M., et al., 2012. Probiotic yogurt improves antioxidant status in type 2 diabetic patients. Nutrition 28 (5), 539–543. https://doi.org/10.1016/j.nut.2011.08.013. Ezcurra, M., Reimann, F., Gribble, F.M., Emery, E., 2013. Molecular mechanisms of incretin hormone secretion. Curr. Opin. Pharmacol. 13 (6), 922–927. https://doi. org/10.1016/j.coph.2013.08.013. FAO, 2001. Probiotics in food. Food Nutr. 85, 71. Ghoshal, U., Shukla, R., Ranjan, P., Ghoshal, U., 2017. 735- expression of TLR-2, 4 and 5 and pro-inflammatory (IL-6, CXCL-11 AND CXCR-3) and anti-inflammatory cytokines (IL-10) among patients with IBS and controls in relation to the gut microbiota. Gastroenterology 152 (5), S161–S168. Gritz, E.C., Bhandari, V., 2015. The human neonatal gut microbiome: a brief review. Front. Pediatr. 3, 17. https://doi.org/10.3389/fped.2015.00017. Gruys, E., Toussaint, M.J.M., Niewold, T.A., Koopmans, S.J., 2005. Acute phase reaction and acute phase proteins. J. Zhejiang Univ. Sci. B 6 (11), 1045–1056. https://doi. org/10.1631/jzus.2005.B1045. Hernández, A., Rull, A., Rodríguez, E., Riera, M., Luciano, F., Camps, J., et  al., 2013. Mitochondrial dysfunction: a basic mechanism in inflammation-related non-­ communicable diseases and therapeutic opportunities. Mediators Inflamm. 1–13. https://doi.org/10.1155/2013/135698. Hill, C., Guarner, F., Reid, G., Gibson, G.R., et  al., 2014. Expert consensus document: the international scientific association for probiotics and prebiotics consensus statement on the scope and appropriate use of the term probiotic. Nat. Rev. Gastroenterol. Hepatol. 11, 506–514. Hirai, S., Takahashi, N., Goto, T., Lin, S., Uemura, T., Yu, R., et al., 2010. Functional food targeting the regulation of obesity-induced inflammatory responses and pathologies. Mediators Inflamm. 367838 (8). https://doi.org/10.1155/2010/367838. Hu, G., Chen, G., Xu, H., Ge, R., Lin, J., 2010. Activation of the AMP activated protein kinase by short-chain fatty acids is the main mechanism underlying the beneficial effect of a high fiber diet on the metabolic syndrome. Med. Hypotheses 74 (1), 123– 126. https://doi.org/10.1016/j.mehy.2009.07.022. Human Microbiome Project Consortium, 2012. Structure, function and diversity of the healthy human microbiome. Nature 486, 207–214. Hunter, D., 2005. Gene–environment interactions in human diseases. Nat. Rev. Genet. 6, 287–298. Icaza, M., 2013. Gut microbiota in health and disease. Rev. Gastroenterol. Mex. 78 (4), 240–248. ILSI, 1999. Scientific concepts of functional foods in Europe. Consensus document. Br. J. Nutr. 81 (1), S1–S27. Jones, S.E., Versalovic, J., 2009. Probiotic lactobacillus reuteri biofilms produce antimicrobial and anti-inflammatory factors. BMC Microbiol. 9, 1–9. https://doi. org/10.1186/1471-2180-9-35. Jones, M.L., Martoni, C.J., Parent, M., Prakash, S., 2017. Cholesterol-lowering efficacy of a microencapsulated bile salt hydrolase-active Lactobacillus reuteri NCIMB 30242 yoghurt formulation in hypercholesterolaemic adults. Br. J. Nutr. 2012, 1505–1513. https://doi.org/10.1017/S0007114511004703. Jumpertz, R., Le, D., Turnbaugh, P., Trinidad, C., Bogardus, C., Gordon, J., Krakoff, J., 2011. Energy-balance studies reveal associations between gut microbes, caloric load, and nutrient absorption in humans. Am. J. Clin. Nutr. 94 (1), 58–65. Kang, H.J., Im, S.H., 2015. Probiotics as an immune modulator. J. Nutr. Sci. Vitaminol. (Tokyo) 61, S103–S105.

480  Chapter 14  Probiotics Beverages: An Alternative Treatment for Metabolic Syndrome

Kanmani, P.K., et al., 2013. Probiotics and its functionally valuable products: a review. Crit. Rev. Food Sci. Nutr. 53 (6), 641–658. https://doi.org/10.1080/10408398.2011.5 53752. Kekkonen, R.A., Lummela, N., Karjalainen, H., Latvala, S., Tynkkynen, S., Järvenpää, S., Korpela, R., 2008. Probiotic intervention has strain-specific anti-inflammatory ­effects in healthy adults. World J. Gastroenterol. 14 (13), 2029–2036. https://doi. org/10.3748/wjg.14.2029. Langa, S., Landete, J.M., Martín-cabrejas, I., Rodríguez, E., Arqués, J.L., Medina, M., 2013. In situ reuterin production by lactobacillus reuteri in dairy products. Food Control 33 (1), 200–206. https://doi.org/10.1016/j.foodcont.2013.02.035. Laparra, J.M., Sanz, Y., 2010. Interactions of gut microbiota with functional food components and nutraceuticals. Pharmacol. Res. 61 (3), 219–225. https://doi. org/10.1016/j.phrs.2009.11.001. Lee, N.K., et al., 2015. Multifunctional effect of probiotic Lactococcus lactis KC24 isolated from kimchi. LWT-Food Sci. Technol. 64 (2), 1036–1041. https://doi.org/10.1016/j. lwt.2015.07.019. Leslie, B.R., 2005. Metabolic syndrome: historical perspectives. Am J Med Sci 330 (6), 264–268. https://doi.org/10.1097/00000441-200512000-00002. Ley, R., Bäckhed, F., Turnbaugh, P., Lozupone, C., Knight, R., Gordon, J., 2005. Obesity alters gut microbial ecology. Proc. Natl. Acad. Sci. U. S. A. 102 (31), 11070–11075. Ley, R., Turnbaugh, P., Klein, S., Gordon, J., 2006. Microbial ecology: human gut microbes associated with obesity. Nature 444 (7122), 1022–1023. https://doi. org/10.1038/4441022a. Li, X., Shimizu, Y., Kimura, I., 2017. Gut microbial metabolite short-chain fatty acids and obesity. Biosci. Microb. Food Health 36 (4), 135–140. https://doi.org/10.12938/ bmfh.17-010. Lumeng, C.N., Saltiel, A.R., 2011. Inflammatory links between obesity and metabolic disease. J. Clin. Invest. 121 (6), 2111–2117. https://doi.org/10.1172/JCI57132. Martínez E, Martín A, Segura M. (2017). Microbiome alterations in people with obesity: the probiotics double role. Diet, Microbiome and Health, Editores: Grumezescu A, Holban A. first ed.. Ed. Academic Press. ISBN: 9780128114407. Mcloughlin, R.F., Berthon, B.S., Jensen, M.E., Baines, K.J., Wood, L.G., 2017. Short-chain fatty acids, prebiotics, synbiotics, and systemic inflammation: a systematic review and meta-analysis. Am. Soc. Nutr. 1–16. https://doi.org/10.3945/ajcn.117.156265. Short-chain. Michel, V., Yuan, Z., Ramsubir, S., Bakovic, M., 2006. Choline transport for phospholipid synthesis. Exp. Biol. Med. 231 (5), 490–504. https://doi. org/10.1177/153537020623100503. Mishra, V., Shah, C., Mokashe, N., Chavan, R., Yadav, H., Prajapati, J., 2015. Probiotics as potential antioxidants: a systematic review. J. Agric. Food Chem. 63 (14), 3615–3626. https://doi.org/10.1021/jf506326t. Mohamadshahi, M., Veissi, M., Haidari, F., Javid, A.Z., Shirbeigi, F.M., 2014. Effects of probiotic yogurt consumption on lipid profile in type 2 diabetic patients: A randomized controlled clinical trial. J. Res. Med. Sci. 19 (6), 531–536. Available from, https:// www.ncbi.nlm.nih.gov/pmc/articles/PMC4155708/. Ng, S.C., Hart, A.L., Kamm, M.A., Stagg, A.J., Knight, S.C., 2009. Mechanisms of action of probiotics: recent advances. Inflamm. Bowel Dis. 15 (2), 300–310. https://doi. org/10.1002/ibd.20602. Nieto-Vazquez, I., Fernández-Veledo, S., Krämer, D.K., Vila-Bedmar, R., Garcia-Guerra, L., Lorenzo, M., 2008. Insulin resistance associated to obesity: the link TNF-alpha. Arch. Physiol. Biochem. 114 (3), 183–194. https://doi.org/10.1080/13813450802181047. Parikh, R.M., Mohan, V., 2012. Changing definitions of metabolic syndrome. Indian J. Endocrinol. Metab. 16 (1), 7–12. https://doi.org/10.4103/2230-8210.91175.

Chapter 14  Probiotics Beverages: An Alternative Treatment for Metabolic Syndrome   481

Plantinga, T.S., van Maren, W.W.C., van Bergenhenegouwen, J., Hameetman, M., Nierkens, S., Jacobs, C., Netea, M.G., 2011. Differential toll-like receptor recognition and induction of cytokine profile by Bifidobacterium breve and lactobacillus strains of probiotics. Clin. Vaccine Immunol. 18 (4), 621–628. https://doi.org/10.1128/ CVI.00498-10. Rani, V., Deep, G., Singh, R., Palle, K., Yadav, U., 2016. Oxidative stress and metabolic disorders: pathogenesis and therapeutic strategies. Life Sci. 148 (1), 183. https:// doi.org/10.1016/j.lfs.2016.02.002. Ridlon, J.M., Kang, D.J., Hylemon, P.B., Bajaj, J.S., 2014. Bile acids and the gut microbiome. Curr. Opin. Gastroenterol. 30 (3), 332–338. https://doi.org/10.1097/ MOG.0000000000000057. Rodríguez, D., Reyes, A., León, P., Sánchez, H., Mosti, M., Aguilar, C., et  al., 2016. Assessment of two different diagnostic guidelines criteria (National Cholesterol Education Adult Treatment Panel III [ATP III] and international diabetes federation [IDF]) for the evaluation of metabolic syndrome remission in a longitudinal cohort of patients undergoing roux-en-Y gastric bypass. Surgery 159 (4), 1121–1128. Rossi, M., Amaretti, A., Leonardi, A., Raimondi, S., Simone, M., Quartieri, A., 2013. Potential impact of probiotic consumption on the bioactivity of dietary phytochemicals. J. Agric. Food Chem. 61, 9551–9558. https://doi.org/10.1021/jf402722m. Sales, N.M.R., Pelegrini, P.B., Goersch, M.C., 2014. Nutrigenomics: Definitions and advances of this new science. J. Nutr. Metab. 2014, 202759. https://doi. org/10.1155/2014/202759. Samuel, B.S., et al., 2008. Effects of the gut microbiota on host adiposity are modulated by the short-chain fatty-acid binding G protein-coupled receptor, Gpr41. Proc. Natl. Acad. Sci. U. S. A. 105 (43), 16767–16772. https://doi.org/10.1073/pnas.0808567105. Sánchez, B., et  al., 2017. Probiotics, gut microbiota, and their influence on host health and disease. Mol. Nutr. Food Res. 61 (1), 1–15. https://doi.org/10.1002/ mnfr.201600240. Slingerland, A.E., Schwabkey, Z., Wiesnoski, D.H., Jenq, R.R., 2017. Clinical evidence for the microbiome in inflammatory diseases. Front. Immunol. 8, 400. https://doi. org/10.3389/fimmu.2017.00400. Sommer, F., Moltzau, J., Bharti, R., Raes, J., Rosenstiel, P., 2017. The resilience of the intestinal microbiota influences health and disease. Nat. Rev. Microbiol. 15, 630–638. Songisepp, E., Kals, J., Kuliisaar, T., Mandar, R., Hutt, P., et al., 2005. Evaluation of the functional efficacy of an antioxidative probiotic in healthy volunteers. Nutr. J. 4–22. https://doi.org/10.1186/1475-2891-4-22. Srinivasan, V.A.R., Raghavan, V.A., Parthasarathy, S., 2012. Biochemical basis and clinical consequences of glucolipotoxicity: a primer. Heart Fail. Clin. 8 (4), 501–511. https://doi.org/10.1016/j.hfc.2012.06.011. Szajewska, H., Guarino, A., Hojsak, I., Indrio, F., et al., 2014. Use of probiotics for management of acute gastroenteritis: a position paper by the ESPGHAN working Group for Probiotics and Prebiotics. J. Pediatr. Gastroenterol. Nutr. 58, 531–539. Tolhurst, G., Heffron, H., Lam, Y.S., Parker, H.E., Habib, A.M., et al., 2012. Short-chain fatty acids stimulate glucagon-like peptide-1 secretion via the G-protein-coupled receptor FFAR2. Diabetes 61 (2), 364–371. https://doi.org/10.2337/db11-1019. Tremaroli, V., 2012. Functional interactions between the gut microbiota and host metabolism. Nature 489, 242–249. https://doi.org/10.1038/nature11552. Ubeda, C., Lipuma, L., Gobourne, A., Viale, A., Leiner, I., et al., 2012. Familial transmission rather than defective innate immunity shapes the distinct intestinal microbiota of TLR-deficient mice. J. Exp. Med. 209 (8), 1445–1456. Waldram, A., Holmes, E., Wang, Y., Rantalainen, M., Wilson, I.D., et al., 2009. Top-down systems biology modeling of host metabotype-microbiome associations in obese rodents. J. Proteome Res. 8 (5), 2361–2375. https://doi.org/10.1021/pr8009885.

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Wang, Y., Wu, Y., Wang, Y., Xu, H., Mei, X., et al., 2017. Antioxidant Properties of Probiotic Bacteria. Nutrients 9 (5), 521. https://doi.org/10.3390/nu9050521. WHO, 2017. Obesity: Preventing and Managing The Global Epidemic. Report of a WHO Consultation. WHO Technical Report Series 894. World Health Organization, Geneva. Young, V., 2017. The role of the microbiome in human health and disease: an introduction for clinicians. Br. Med. J. 356, https://doi.org/10.1136/bmj.j831. Yukio, K., Sato, M., Ogawa, A., Miyoshi, M., Uenichi, H., et al., 2013. Effect of Lactobacillus gasseri SBT2055 in fermented milk on abdominal adiposity in adults in a randomised controlled trial. Br. J. Nutr. 110 (9), 1696–1703. https://doi.org/10.1017/ S0007114513001037.

Further Reading Gibson, G.R., Nicholson, J.K., 2009. Top-down systems biology modeling of host metabotype microbiome associations in obese rodents. J. Proteome Res. 8 (5), 2361– 2375. https://doi.org/10.1021/pr8009885.

A NEW GENERATION OF PROBIOTIC FUNCTIONAL BEVERAGES USING BIOACTIVE COMPOUNDS FROM AGRO-INDUSTRIAL WASTE

15

Dan Cristian Vodnar⁎,†, Lavinia Florina Călinoiu⁎,†, Laura Mitrea⁎,†, Gabriela Precup⁎,†, Maria Bindea⁎,†, Andrea Mariana Păcurar⁎,†, Katalin Szabo⁎,†, Bianca Eugenia Ştefănescu†,‡ ⁎

Faculty of Food Science and Technology, University of Agricultural Sciences and Veterinary Medicine Cluj-Napoca, Cluj-Napoca, Romania, †Institute of Life Sciences, University of Agricultural Sciences and Veterinary Medicine ClujNapoca, Cluj-Napoca, Romania, ‡Department of Pharmaceutical Botany, Iuliu Hațieganu University of Medicine and Pharmacy, Cluj-Napoca, Romania

15.1 Introduction The market of functional beverages is continuously increasing so the key research priority in this field is to develop new products rich in bioactive compounds targeting lifestyle diseases. The development of new probiotic nondiary functional beverages with an inflated demand from the lactose-intolerant and vegetarian population is in vogue. Clinical studies have been done in order to test the efficacy of these probiotic strains. Moreover, the valorizing of Romanian agro-industrial wastes is another objective of this research, due to the fact that they contain highvalue natural compounds which may have health-promoting characteristics and their disposal may harm the environment by contributing to pollution. The presence of bioactive molecules, such as fatty acids and phenolic compounds, in agro-industrial waste makes fruit and vegetable leftovers more valuable for the food industry. Fruits and vegetables contain bioactive compounds that impart health benefits beyond basic nutrition (Oomah and Mazza, 2000). These wastes are rich in bioactive compounds and can thus be improved and incorporated into food supplements (European Commission Final Report, 2010). Functional and Medicinal Beverages. https://doi.org/10.1016/B978-0-12-816397-9.00015-7 © 2019 Elsevier Inc. All rights reserved.

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15.2  Functional Beverages Market 15.2.1  The Concept of Functional Food Since the early 21st century, our society has been witnessing a continuous growth in life expectancy and, at the same time, greater attention to quality. Consumers are increasingly concerned about their health and are paying extra attention to their lifestyle and the overall healthiness of their eating habits (Szakály et al., 2012) hence their demands in the field of food production has changed considerably. In this context, it has been observed a growing recognition of the key role of foods and beverages in disease prevention and treatment, since consumers more and more believe that what they eat reflects directly on their health. As a consequence, the focus of scientific research has moved from the primary function of food as the source of energy and nutrients to the more subtle action of biologically active food components having the ultimate function to improve the physical and mental well-being of the consumers. As a result, a new term—functional food—was proposed (Granato et al., 2010). However, the idea of health-promoting foods is not new: Hippocrates wrote 2400 years ago “Let food be thy medicine and medicine be thy food” (Otles and Cagindi, 2012), and the concept of functionality of food products and herbs was a commonplace for the Asian communities (Valls et al., 2013). Nonetheless, the term “functional food” itself was first used in Japan, in the mid-1980s, and later on became a concept, that was first promoted by the Japanese scientists who studied the relationships between nutrition, sensory fulfilment, fortification, and modulation of physiological systems, resulting in the distinction of a specific health-related food category called FOod for Specified Health Uses (FOSHU) (Lau et al., 2012). The Japanese Ministry of Health, Labor, and Welfare defined FOSHU as “foods which are expected to have certain health benefits, and have been licensed to bear a label claiming that a person using them for specified health use may expect to obtain the health use through the consumption thereof” (Bigliardi and Galati, 2013). There is no doubt that the Japanese interest in functional foods has also brought awareness for the need of such products in places like Europe and the United States (Siro et al., 2008). A large number of definitions currently exist for the term “functional foods” (Bigliardi and Galati, 2013), however, none of them is universally accepted due to the fact that functional foods are, perhaps, more accurately viewed as a concept and a marketing terminology than a well-defined group of food products (Ashwell, 2002; DelgadoAndrade, 2017; Roberfroid, 2002). Moreover, in most countries, there is

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no legislative definition of the term and making a distinction between conventional and functional foods is challenging even for nutritionists and food experts (Niva, 2007). In Europe, the interest in functional foods started in the latter half of the 1990s. The European Commission initiated an activity called Functional Food Science in Europe (FuFoSE) to examine the concept of functional foods through a science-based approach. As a result, the European Commission advocated that “a food product can only be considered functional if together with the basic nutritional impact it has beneficial effects on one or more functions of the human organism thus either improving the general and physical conditions or/and decreasing the risk of the evolution of diseases” (Ozen et al., 2012; Pravst, 2012). Therefore, a functional food could be: a plain natural food; a food in which a component has been upgraded through special growing conditions, breeding, or biotechnological means; a food to which a component has been added to provide benefits; a food from which a component has been removed by technological or biotechnological means so that the food provides benefits otherwise nonexistent; a food in which a component has been replaced by an alternative component with complementary properties; a food in which a component has been customized by enzymatic, chemical, or technological means to provide a benefit; a food in which the bioavailability of a component has been modified; or a combination of any of the above (Pravst, 2012). Functional foods are the first to bear health claims. According to the Codex Alimentarius, a claim means “any representation which states, suggests or implies that a food has particular characteristics relating to its origin, nutritional properties, nature, production, processing, composition, or any other quality.” Health claims are the key factors for the expansion of the functional food market (Corbo et al., 2014). In December 2006, the Council and Parliament adopted the Regulation 1924/2006 on nutrition and health claims made on foods. Without precedent, this Regulation lays down harmonized rules across the European Union for the use of nutrition claims such as “low fat,” “high fiber,” or health claims such as “reducing blood cholesterol” (European Parliament and Council, 2006). Functional foods represent one of the most interesting areas of research and innovation in the food industry and their development is a key research priority for food design and a challenge for both industry and science sectors (Annunziata and Vecchio, 2011). Their objectives are numerous: they improve the general conditions of the body (e.g., pre- and probiotics), decrease the risk of some diseases (e.g., ­cholesterol-lowering products), and could be used for curing some ailments (Mark-Herbert, 2004). Overall, functional foods add value to the products that are otherwise basic, they individualize products that would otherwise be

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commodities, they appeal to a basic human motive—health—and the presumption was that they would lead markets that grow, if not in volume, but in value (Grunert, 2010).

15.2.2  Functional Beverages Categories Currently, the range of functional foods includes products such as baby foods, dairy foods, confectionery, ready meals, baked goods and cereals, snacks, spreads, meat products, and beverages (Ofori and Peggy, 2013). In particular, beverages are by far the most effective functional food category due to several reasons: convenience and possibility to meet consumer requirements for container contents, size, shape, and appearance, as well as ease of delivery and storage for refrigerated and shelf-stable products. Moreover, they are an excellent delivering means for nutrients and bioactive compounds counting prebiotics, probiotics, vitamins, minerals, antioxidants, ω-3 fatty acids, plant extracts, and fiber (Corbo et al., 2014). The different types of commercially available products could be clustered as follows: dairy-based beverages including probiotics and minerals-/ω-3-enriched drinks, vegetable and fruit beverages, and sports and energy drinks (Corbo et al., 2014). Functional foods are guided notably by a main sector: dairy. The reasons of this choice by consumers are mainly due to the strong dairy tradition of many countries, but also to the attention paid to these products (Vicentini et  al., 2016). Within dairy beverages, fresh milk, fermented milk, and yogurt drinks are the most common products, as they are considered as excellent transporters for probiotics, bioactive compounds, vitamins, and minerals (Corbo et al., 2014; Vijaya Kumar et al., 2015). However, with an increase in the consumer vegetarianism throughout the developed countries, there is also a demand for the vegetarian functional products. Furthermore, lactose intolerance and the cholesterol content are two major drawbacks related to the fermented dairy products, granting nondairy beverages a big importance, worldwide (Granato et al., 2010). In recent years, plant sources (cereals and legumes) are accepted as functional food and nutraceuticals due to the presence of health-promoting components such as dietary fibers, minerals, vitamins, and antioxidants (Das et al., 2012). It has been recently indicated that fruit juices could be ideal media for probiotic due to their content of essential nutrients (Granato et al., 2010). Some examples of novel functional beverages are whey-based prickly pear (Baccouche et al., 2013) and grape-based beverages (Di Cagno et al., 2010), cereal-based probiotic drink (Rathore et al., 2012), fruit-­beverages (Gad et al., 2013; Gunathilake et al., 2013), and some vegetable beverages (Awe et al., 2013; Gironés-Vilaplana et al., 2012).

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In today’s world, beverages are no longer considered simply as thirst-suppressors; consumers look for specific functionality in these drinks, which form a part of their lifestyle. Functionality in these beverages could be to address different needs and lifestyles—to boost energy, fight aging, fatigue and stress, and target-specific diseases and the sector is still spreading (Sethi et al., 2016). Hence, there is a continuous interest by the functional beverage industry to launch new beverage products that would be similarly attractive to the consumers and would target the desired requirements.

15.2.3  Market Evolution Functional food development engages several distinct stages from concept to successful market implementation. In particular, consumer acceptance has been recognized as a key factor to successfully debate market opportunities (Siro et al., 2008). Due to the lack of an internationally accepted definition of functional food, researchers stated that it is very difficult to study the worldwide functional food markets (Corbo et  al., 2014; Ozen et  al., 2012). However, despite the shortfall of precision concerning the data of global sales, functional foods have undeniably been reported as the top trend in the food industry (Valls et al., 2013). Japan is a leader in the field, followed by the United States and Europe (Bleiel, 2010). It is not surprising that in Japan this market is significant due to the fact that it is regarded as the birthplace of functional food: only between 1988 and 1998, more than 1700 functional food products have been launched there (Bigliardi and Galati, 2013). As more recently reported, the global launches of functional products between 2005 and 2009 have more than doubled, from 904 to 1859. Between 2008 and the first half of 2009, the United States was the forerunner in “healthy” product launches (881 products), followed by Japan (314), Italy (325), the United Kingdom (237), Germany (235), and France (150) (Sorenson and Bogue, 2009). These products represent a sustainable trend in the food market. In particular, the functional food market is growing worldwide, as demonstrated by the fact that new functional products are being launched continuously (Bigliardi and Galati, 2013). As previously stated, the main market for functional foods is Asia Pacific. Recent studies evaluated that the revenues for the field of functional foods in Asia and the Pacific islands constitute as much as 34% of total revenue worldwide. The second largest market in the world is the North American one, essentially consisting of the United States and Canada. These two countries alone reach 25% of total ­revenue worldwide. Particularly, in the United States, the field of functional foods is considered a real business (Vicentini et al., 2016).

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As far as Europe is concerned, the demand for functional foods varies considerably from country to country. The market development has been relatively stable and profitable, with a 16% of total revenue worldwide. The main source of revenue is the United Kingdom with 20% of total revenues which corresponds to about US$ 7.4 billion, followed by Germany with 14% and France with 13% of total revenues, which corresponds both to about US$ 4.9 billion. Spain and Italy account for 12% and 11% of total revenues, respectively, which correspond to about US$ 3.7 billion. In Latin America, the functional foods and food production are relatively “new,” but very promising. Lastly, the markets of Australia, New Zealand, Africa, the Middle East, and Eastern Europe can be considered negligible, since all together they make up just 8% of total revenue worldwide. Based on the studies conducted so far, it can be concluded that these products have had an immediate success in most of the industrialized countries as well as in the developing ones, and it has been highlighted that the main trends that drive the success of functional foods could be identified as: age, sex, education, and demographic changes (Urala and Lähteenmäki, 2007), the restraint of health-care costs (Milner, 2000), media, increased information accessibility, nutrition labeling (Kotilainen et al., 2006), increasing emphasis on healthy diet and global prevention of chronic diseases, innovations in food technology and brand differentiation, and greater emphasis on value for money (Vicentini et al., 2016). The future of functional beverages depends on the unequivocal demonstration of their efficacy in promoting health. Thus, a joint venture between food producers and researchers is advisable, as a tool to provide scientific evidence of many health claims, as well as a way to find successful strategies to improve the appeal of functional beverages (Corbo et al., 2014).

15.3  Health Benefits of Functional Beverages The emerging trend in the functional beverage market is to design new beverages that can meet the changing needs and the present demands of the consumer, by offering alternatives of health beverages. In today’s globalized world, the growing urbanization, lifestyle changes, and dietary habits can have negative impacts on human health, therefore, the consumer became more health conscious and aware of the possible beneficial effects of functional foods (Puiggròs et al., 2017). Functional beverages are gaining more and more acceptance, being the first choice for grab-and-go foods among consumers, who are looking for particular functional effects like energy boosting, ­counteracting

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stress, fatigue, or targeting specific illnesses. Therefore, the food industry is continuously focusing on the designing innovative and functional beverages which can satisfy the consumer’s requirements. A major development within this field pertains to beverages containing prebiotic and probiotic bacteria, as health-promoting substances, designed to improve gut flora. This segment of the functional beverages market is increasing in popularity considering the fact that probiotics are live microorganisms that can exert beneficial effects on the intestinal flora, if administered in adequate amounts (Hill et  al., 2014). The major interest now is to create novel probiotic formulations in order to tackle health problems as lactose intolerance and milk protein allergy, gastrointestinal disorders like irritable bowel syndrome (IBS), inflammatory bowel disease, or control cholesterol levels, so as to alleviate the disadvantages of diary-based beverages (Stanton et al., 2005). Therefore, researchers have identified alternative matrices that could serve as carriers for probiotic bacteria, like cereals, fruit, and vegetable juices, or even by-products from the agro-food industry, which represent sustainable and economically friendly substrates (Kandylis et al., 2016).

15.3.1  Irritable Bowel Syndrome Within the last decade, there is a growing body of evidence highlighting the effects of probiotic bacteria in the management of gastrointestinal disorders, such as IBS. IBS is one of the most common gastrointestinal disorder in the industrialized world, with a high prevalence of 11.2%, based on a meta-analysis and a continuously increasing incidence, 1%–2% annually (Chey et  al., 2015; Lacy et  al., 2016). IBS could be defined as recurrent abdominal pain, with a frequency of at least once a week and associated with two or more of the following: changes in stool frequency, changes in appearance of the stool, or related to defecation, as updated according to the Rome IV criteria for IBS (Palsson et al., 2016). For diagnosis, other diseases with a common symptomatology are excluded, considering the fact that there is no specific biomarker. IBS is classified into three main subtypes according to the predominant disorder in bowel habits: patients who tend to have predominant diarrhea, patients who tend to have predominant constipation, or those with mixed bowel habits (Lacy, 2016). Women are more prone to have IBS than men and young people are more affected than those older than 50 years old (Lacy et al., 2016). IBS could be presented as a multifactorial disease, as the etiology is not yet known, several triggers factors being guilty for the expression of the disease, like several foods, stress, lifestyle, gastrointestinal infections, alterations in the intestinal flora, low-grade inflammation,

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visceral hypersensitivity, and genetic factors, varying from patient to patient (Zhuang et al., 2017). Among the accused foodstuffs, we could mention: milk and dairy products, pastries, coffee, cabbage, onions, beans, spicy spices, roasted, and smoked products (Böhn et al., 2013). There are several treatment options for IBS, but the probiotic approach seemed to be promising as there is a valuable body of evidence that certain living organisms, when ingested in an adequate amount, can confer health benefits to the most, by modulating the gut microbiota (Hill et al., 2014). In order to be effective, probiotics must fulfil several requirements as to be nonpathogenic, nontoxic, viable in a certain minimum level as to exert the beneficial effects in the gut, but also to maintain their viability during different stressful conditions such as under environmental factors (variations in temperature, pH, oxygen, and humidity), production and storage, as well as the exposure to the acidic conditions from the stomach (Hill et al., 2014; Calinoiu et al., 2016; Yao et al., 2017). Their ability to survive to all these conditions is a key factor for providing the health benefits, albeit probiotics could lose their viability during production, storage as well as through their passage in the gastrointestinal tract (Sarao and Arora, 2017). Therefore, different strategies had to be developed in order to protect them from the above mentioned factors, by using the encapsulation methods and incorporate them in carefully designed microcapsules (Vodnar et al., 2010; Vodnar and Socaciu, 2014; Yao et al., 2017). The microencapsulation methods of different probiotic bacteria will be extensively presented within this chapter.

15.3.2  Clinical Studies for IBS Over the past decades, the number of research studies focusing on the dietary management of IBS by administering probiotics increased dramatically. The probiotics used in the clinical trials were mainly lactobacilli and bifidobacteria, in a single strain or in combination with other strains, in different dosages. A series of meta-analyses pooled together the studies evaluating different probiotic species/strain and the latest studies from 2016 and 2017 tackling probiotic intervention in IBS are presented in Table 15.1. To exemplify, a probiotic beverage containing 400 mL of fruit juice fortified with 5 × 107 UFC/mL of Lactobacillus plantarum 299v, revealed a decrease in the frequency of abdominal pain and bloating in patients with IBS symptoms, after 4 weeks of intervention (Niedzielin et  al., 2001). Moreover, another clinical study from 2017 tested the efficacy of a double-coated supplement containing. Five billion bacteria per capsule of multispecies probiotics (Lactobacillus acidophilus, L. plantarum, Lactobacillus rhamnosus, Bifidobacterium breve, Bifidobacterium lactis, Bifidobacterium longum, and Streptococcus

Table 15.1  The Effect of Different Probiotic Bacteria in Irritable Bowel Syndrome Study Type

Intervention

Patients

Outcome

Reference

Randomized ­double-blind controlled trial

Dual-coated or noncoating probiotic supplement (Duolac Care): 5 × 109 Lactobacillus acidophilus, L. plantarum, L. rhamnosus, Bifidobacterium breve, B. lactis, B. longum, and Streptococcus thermophiles, two capsules, twice/day

50 IBS-D

Han et al. (2017)

Randomized ­double-blind controlled trial

Capsule with: Group 1: 5 × 109 CFU/d L. acidophilus PBS066; 5 × 109 CFU/d L. reuteri PBS072; Group 2: 5 × 109 CFU/d L. plantarum PBS067; 5 × 109 CFU/d L. rhamnosus LRH020; 5 × 109 CFU/d B. animalis spp. lactis BL050. 60 days, 30 days follow-up

150 IBS-C patients

Significant improvement in overall discomfort in the dual-coating group and the ratio of normal stools to hard or watery stools had a better effect from dual-coated probiotics compared to noncoated probiotics The overall symptoms improved (pain, flatulence, and bloating), improvements in the quality of life The numbers of Lactobacillus spp. and Bifidobacterium spp. of the mixtures increased and all the probiotic strains remained in the gut also 30 days after the follow-up from the last intake, except for Bifidobacterium

Mezzasalma et al. (2016)

Continued

Table 15.1  The Effect of Different Probiotic Bacteria in Irritable Bowel Syndrome—cont’d Study Type

Intervention

Patients

Outcome

Reference

Randomized ­double-blind controlled trial

Capsule with 11 different strains: 3 × 109 CFU/d L. rhamnosus LR5; 2 × 109 CFU/d L. casei LC5; 1 × 109 CFU/d L. paracasei LPC5; 1 × 109 CFU/d L. plantarum LP3; 5 × 109 CFU/d L. acidophilus LA1; 4 × 109 CFU/d B. bifidum BF3; 1 × 109 CFU/d B. longum BG7; 2 × 109 CFU/d B. breve BR3; 1 × 109 CFU/d B. infantis BT1; 2 × 109 CFU/d S. Thermophiles ST3; 3 × 109 CFU/d L. bulgaricus LG1 3 × 109 CFU/d Lactococcus lactis SL6. Twice daily for 8 weeks B. longum NCC3001 Unclear dose 6 weeks

107 IBS-D patients

Symptoms improved, but no difference between the probiotic and the placebo groups in pain intensity’s tool consistency or overall responder rates

Hod et al. (2017)

44 IBS-D and IBS-M patients with mild-to–moderate anxiety or depression

No effect on gut symptoms, but beneficial effects on mild-to-moderate comorbid depression

Pinto-Sanchez et al. (2017)

Randomized ­double-blind controlled trial

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thermophiles) on the patients with diarrhea-predominant IBS, and after 4 weeks the symptoms improved (Han et al., 2017). A meta-­analysis recently revealed that multispecies probiotics with Bifidobacterium infantis strains considerably reduced the abdominal pain and bloating, but the treatment with the single probiotic B. infantis did not have a positive impact on IBS symptoms (Yuan et al., 2017). In addition, another mix of probiotic formulation containing lactobacilli and bifidobacteria highlighted the efficiency in IBS-C patients, who received two different formulation of probiotics once daily for a period of 60 days and were followed up for a further period of 30 days. The severity of symptoms decreased (pain, flatulence, and bloating), and the improvements in the quality of life were significant when comparing to placebo, with maintained effects even after 30 days of the last intake (Mezzasalma et al., 2016). Conversely, the study of Hod, Sperber et al. showed no significant superiority of a probiotic treatment with lactobacilli and bifidobacteria in IBS-D patients. Similarly, Pinto-Sanchez et al. did not find an improvement in gut symptoms associated with IBS after intake of B. longum, but beneficial effects on depression were observed. Therefore, the current evidence suggests a potential beneficial effect of specific probiotic strains in certain IBS symptoms, but the studies have their limitations in terms of number of patients, loss of follow-up, exact knowledge of the optimal strain, dose, and duration of treatment with longer beneficial effects.

15.3.3  Lactose Intolerance Syndrome Lactose is a type of natural sugar found in milk, breast-milk, and dairy products. This disaccharide is broken down in the small intestine into glucose and galactose under the action of enzyme lactase. Lactose intolerance is a gastrointestinal disorder with symptoms caused by decreased ability to digest lactose. Most often symptoms may include bloating, abdominal pain, cramps, gas, and diarrhea installing between 30 min and 2 h after milk consumption. Severity of the symptoms depends on the amount of lactose intake although a substantial variability in response can be expected as the manifestation of lactose intolerance varies among individuals. The development of the symptoms depends on several individual factors like diet, fermentation capacity of the gut flora, oro-cecal transit time, and psychological factors (Roškar et al., 2017). Lactose intolerance is a consequence of lactase deficiency in the lining of duodenum. Terms like “lactose malabsorbtion” or “lactase nonpersistence” is referring to the condition of impaired production of the enzyme β-galactosidase (lactase). Hypolactasia indicates insufficient lactase level in the organism while alactasia illustrates the

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complete absence of lactase. This fact conducts to the individuals who cannot digest any milk products and people who can eat or drink small amounts of products containing lactose without problems. This type of intolerance occurs in modest proportion in European descent, Scandinavian population manifesting the lowest lactose intolerance in the world (Vuorisalo et  al., 2012) on contrary in some communities from Asia >90% of adults are influenced of lactose malabsorbtion, comprising the highest incidence. Lactose intolerance can be defined as sensitivity to dairy products caused by lactase deficiency and it is not an allergy. Milk allergy occurs in only 4% of the population and it manifests through an immune reaction when milk proteins are present in the organism. The principal symptom of lactose intolerance is an adverse reaction to the products containing lactose, evidenced by a general abdominal discomfort with specific manifestations: abdominal pain, flatulence, rumble, diarrhea, nausea, and/or vomiting, the intensity of these symptoms being directly proportional with the amount of lactose consumed. According to its causes, lactose intolerance is classified in three categories. Primary lactase deficiency or primary hypolactasia has genetic origins and it is caused by the absence of a lactase persistence allele. A genetic mutation developed in the human organism several thousand years ago activated LCT gene—responsible for lactase production—maintaining it functional even after breast feeding is stopped. This phenomena called lactase persistence swept through Europe about 4300 years ago (Gibbons, 2015). Lactose intolerant people do not have this genetic mutation in their DNA, thereby less lactase is produced by the organism overtime, leading to hypolactasia in adulthood. Congenital lactase deficiency is a very rare form of lactose intolerance manifested from birth by a complete lack of lactase. This type of disorder is inherited from parents and it blocks the expression of lactase enzyme making lactose digestion impossible. Secondary hypolactasia refers to an injury to the small intestine and in general it is not a permanent condition. The causes of secondary lactase deficiency could be ulcerative colitis, chemotherapy, acute gastroenteritis, celiac disease, infections, intestinal parasites, or other causes that might injure the small intestine. Usually, by treating the underlying cause of secondary hypolactasia the lactose intolerance can be reversible. The bacterial fermentation of undigested lactose in the colon and osmotic processes provokes symptoms of lactose intolerance although diseases like IBS, celiac disease, or inflammatory bowel disease may produce similar symptoms. In order to diagnose properly, medical tests along with physical exams and medical, family and diet history is required. The self-reported symptoms after ­lactose ­ingestion could

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be highly susceptible of a subjective perception, therefore, medical tests can provide more accurate results. Medical tests for lactose intolerance diagnosis can include hydrogen breath test, blood test, stool acidity test, intestinal biopsy, or a noninvasive molecular marker test. Most commonly used tests are hydrogen breath test and stool acidity test. The first method detects the amount of hydrogen in the exhaled air by a person who ingested a known amount of lactose. In the consequence of undigested lactose, high amount of hydrogen is produced by a lactose intolerant patient. Yet, the sensitivity of this test has been evaluated as suboptimal (Roškar et  al., 2017). Stool acidity test is mostly used in infants with lactose intolerance suspicions as it represents a facile and noninvasive detection method. The undigested lactose mixed with the bacteria from the colon causes acidity in stools, pH value being situated below 5.5 in case of lactose intolerance. Milk and milk products are the major sources of calcium and other nutrients necessary for an equilibrated diet. Lactose intolerant people may not get enough calcium or vitamin D which may predispose them to decreased bone accrual, osteoporosis, and other adverse health outcomes, even though, in most cases, individuals do not need to eliminate dairy consumption completely (Suchy et al., 2010). Scientific work demonstrated that the equivalent of one cup of milk consumed together with other foods can be tolerated by individuals with lactose malabsorbtion without significant symptoms. However, dairy foods are an excellent source of calcium, magnesium, potassium, protein, and vitamin D, these nutrients are available in other foods and supplements. In the management of lactose intolerant diagnosed individuals, there are no definitive recommendations and more scientific research is needed on potential therapeutic interventions. Individualized strategies could combine colonic adaptation, where lactose intake is increased gradually in time, with lactase-treated products or alternative nutrient sources to overcome negative effects of milk and milk products absence from the diet. A promising direction of therapeutic approaches proposed could be represented by probiotic supplementation (Almeida et al., 2012). Probiotics are microorganisms providing health benefits and may be an option for the improvement of lactose intolerance-related symptoms. By modulating the composition and metabolic activities of the colonic microbiota with different bacterial strains may help to alleviate lactose intolerance (Roškar et al., 2017). People diagnosed with lactose intolerance might develop other health outcomes because of dairy exclusion diets. The lack of calcium can affect the growth and development of skeleton, leading to the risk of osteoporosis and fragility fractures in adulthood. In order to

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prevent secondary affections of lactose intolerance and to temper the unpleasant symptoms, more scientific work is necessary in therapeutic interventions evolvement.

15.4  Probiotic Bacteria The importance of the gastrointestinal microbiota was efficiently highlighted by recent findings in the field of biology and microbiology, and by different clinical studies, through pointing out that the gut population regulates the human health and the appearance of diseases (Anselmo et  al., 2016). Thereby, the structure of the gastrointestinal microbiota can be influenced and modified by the administration of probiotics, impacting the cure process of an impressive number of illnesses, by strengthening and stimulating the immune system (Mitrea et al., 2017; Ștefănuț et al., 2015; Vodnar and Socaciu, 2014). Regarding the antimicrobial activity of probiotic strains, especially of some lactic bacteria, it has been shown that most of them present inhibitory potential over the pathogenic strains, mainly on the Gram-positive strains, but also on the pathogenic yeast strains, too. So, it is considered that the probiotic bacteria can be used in the reestablishment of the intestinal microflora (Sârbu et al., 2015). In general terms, the probiotics can be named as viable microorganisms which are able to promote and to support a helpful balance of bacteria to live in the gastrointestinal tract (Holzapfel et al., 2001). The good functioning of gut microbiota is influence by the presence of probiotics through a complex network of events which can also influence even the nervous system mechanisms, causing the appearance and the progress of mood disorders like depression or anxiety (Grant and Baker, 2017). The administration of probiotics could also ease the gas and the bloating produced by other digestive disorders, including inflammatory bowel disease, chronic stomach inflammation, or ulcers. Several studies have shown plausible benefits for reducing symptoms of intolerance to lactose, which can include gas and bloating. These affections can be avoided by using certain strains of probiotics (Holzapfel et al., 2001).

15.4.1  Lactobacillus casei L. casei—is one of the numerous bacteria species belonging to the genus Lactobacillus, and it is a well-known probiotic strain for its beneficial impact on human health (Cai et al., 2007). L. casei strain can be isolated from different niches, like raw or fermented dairy products, intestinal tracts or reproductive systems of humans and animals, and fresh and fermented plant products

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(Cai et  al., 2007). L. casei synthesize lactic acid through fermentation processes, a very important property due to the fact that L. casei can be used for multiple purposes, as: manufacturing of cheeses and yogurts, reducing the cholesterol levels, enhancing the immune system’s response, and inhibiting the intestinal pathogens colonies, it can be used in the diarrheal episodes control, it can ease the lactose intolerance, and it can be successfully used as a probiotic strain for food applications like functional and nondairy beverages (Borşa et al., 2015; Mishra and Prasad, 2005; Vodnar et al., 2012). Actually, L. casei has been extensively connected with a number of gastric and intestinal benefits that can apparently be acquired only by this organic channel of the probiotic type. For example, there are several studies which are presenting the L. casei’s ability to adhere to the mucosal layer of the gastrointestinal tract. Besides, these colonies are capable to survive to multiple digestive cycles and also to contribute to the proteins synthesis (Holzapfel et al., 2001; Oozeer et al., 2002). L. casei is presented as a very efficient metabolizer of milk sugars, which means that the quality of life of people suffering from lactose intolerance could be even more improved (Trani et al., 2017; Vodnar et al., 2012). L. casei has been connected to the improvement of more serious intestinal disorders like Crohn’s disease and IBS, by reducing the intestinal inflammation, by normalizing the bowel motility patterns, and by ameliorating the symptoms commonly generated by these intestinal imbalances. The cases of diarrheal episodes, mostly in children, have similarly been shown to be improved under L. casei administration (Sarjapuram et al., 2017).

15.4.2  Lactobacillus plantarum L. plantarum strain is one of the most important and versatile species of its group, and it can be defined as being part of the microbiota of starchy foods, cereals, meat, dairy products, vegetables, fruits, and beverages (Da Silva Sabo et al., 2014). Various strains of L. plantarum have been isolated from various ways such as fermented milk, cheeses, pickled cucumbers, olive oils, pineapple, grapefruit, sorghum, beer, and kefir. These strains have been shown to be able to cross the gastric juice barrier and reach the intestine surface, and for this reason, they are considered as natural members of the intestinal microflora (Da Silva Sabo et al., 2014). L. plantarum strains are presented as saprophyte microorganisms, usually associated with plants and fermentative materials, playing an important role in the preservation of fermented products due to their ability to synthesize antimicrobial substances such as diacetyl and hydrogen peroxide (Daeschel et al., 1990). Some authors suggest that L. plantarum possess the ability to adapt to different environment conditions because of their capacity

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to ferment various carbon sources. L. plantarum is frequently used in starter cultures in the fermentation process of foods, and it is also considered as a basic ingredient of probiotic foods; the strain of L. plantarum 299V is one of the most commonly marketed model (Da Silva Sabo et al., 2014). Literature studies underline that L. plantarum strains, like S25, S2-6, and S4-1 are very tolerable with the gastrointestinal transit (Table 15.2), having a strong resistance at low pH values (Table 15.3) and an impressive supportability toward bile secretions—oxgall (Table 15.4) (Yu et al., 2013). The Lactobacillus antimicrobial potential is mainly due to the organic acids production (i.e., lactic acid, acetic acid, and succinic acid), and to the specific bactericidal metabolites

Table 15.2  The Viability of L. plantarum Strains in the Presence of Gastric and Intestinal Substances (log CFU*) (Yu et al., 2013) The Number of Viable Cells in the Presence of Simulated Gastric Juice: Pepsin 3 mg/mL

The Number of Viable Cells in the Presence of Simulated Intestinal Juice: Pancreatin 1 mg/mL

Strains

0 min

pH 3.0 (180 min)

0 min

pH 8.0 (240 min)

S2-5 S2-6 S4-1

8.88 8.85 8.88

4.01 5.23 3.48

8.79 8.74 8.97

8.65 8.69 8.82

Table 15.3  The Viability of L. plantarum Strains at Different pH Values (log CFU*) (Yu et al., 2013) pH 2.0

pH 3.0

Strains

0 min

60 min

0 min

60 min

180 min

S2-5 S2-6 S4-1

7.70 7.80 8.10

5.59 5.79 5.99

7.80 7.80 8.00

6.89 7.69 7.80

6.71 7.50 7.70

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Table 15.4  The Viability of L. plantarum Strains in the Presence Different Concentrations of Bile Juice— Oxgall (log CFU*) (Yu et al., 2013) 240 min Strains

0 min

0.3% oxgall

0.5% oxgall

1.0% oxgall

S2-5 S2-6 S4-1

8.80 8.80 8.40

8.50 8.50 8.00

8.60 8.40 7.80

7.70 8.30 7.60

(plantaricin, acidophylin, lactocin, hydrogen peroxide, diacetyl, etc.) of each individual strain (Bajaj et al., 2015; Milioni et al., 2015). L. plantarum strains exert a strong antifungal activity too, by presenting a very large inhibitory area against molds (i.e., Penicillium expansum and Aspergillus parasiticus), after 5 days of incubation on Petri dish (Luz et al., 2017). The gastrointestinal disturbances, in most cases, are generated by the overpopulation of the gut microflora with enteric pathogens (i.e., Escherichia coli), which are first responsible for diarrheal episodes among the world (Kumar et al., 2016; Presterl et al., 1999). By incorporating probiotic strains in new functional foods or vegetable beverages, the well-being of human health could efficiently be improved.

15.4.3  Bifidobacterium infantis B. infantis—a “friendly” lactic acid bacteria belonging to the Lactobacillus group as well. It is naturally found in the oral cavity and in the gastrointestinal tract. B. infantis is one of the beneficial strain which maintain a healthy digestive tract (Imperatore et  al., 2017; Vodnar and Socaciu, 2012). Some evidence suggests that taking probiotics strains like B. infantis could help in treating certain health conditions as IBS, by reducing the gas and bloating (Allen et al., 2017; Yuan et al., 2017). Several studies on animal models have reported in “Gut Microbes” Journal that the B. infantis strain significantly ameliorated the symptoms associated with ulcerative colitis, chronic fatigue syndrome, and psoriasis, by reducing the abdominal pain and inflammation. The particular strain of B. infantis 35624 exerts beneficial immunoregulatory effects by mimicking commensal-immune interactions. It was

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demonstrated on clinical trials that B. infantis 35624 has the ability to reduce the systemic pro-inflammatory biomarkers in both gastrointestinal and nongastrointestinal conditions (Groeger et al., 2013). The probiotics influence various mediators throughout the braingut axis, fact that was demonstrated with a specific strain of B. infantis that lowered the levels of pro-inflammatory cytokines in animal models tested for IBS symptoms. It was also noticed that the same strain has the potential to increase the level of polyunsaturated fatty acids, and showed antiinflammatory effects when it was administrated with α-linoleic acid compared with α-linoleic acid administration alone (Saulnier et al., 2013). B. longum subsp. infantis ATCC 15697 presents a good survival rate and viability at different gastrointestinal conditions when it is administered encapsulated with alginate-poly-l-lysine-alginate to the human gut microbiota. The alginate layer protects the probiotic strain against gastric acid and bile insults, besides, the microencapsulation increase the B. infantis ATCC 15697 cells survival at various pH values (Rodes et al., 2014). B. longum spp. infantis 35624 has been proved to efficiently survive transit through the gastrointestinal tract and to exert potent antiinflammatory effects in vivo (in animal models). Additional homeostatic effects, like the enhancement of gut barrier function have also been demonstrated, moreover, this microorganism has been shown to accelerate the intestinal transit and to diminish the visceral hypersensitivity. Therefore, clinical trials have mainly been focused on the IBS where consistent and clinically eloquent benefits have been demonstrated (Quigley, 2017).

15.4.4  Bifidobacterium breve B. breve—a bacterial strain mostly found in the human breast milk, in the gut flora of infants, but it is also found within the fully grown human intestinal microflora. This strain develops a symbiotic relationship with their host by manipulating their unique metabolic pathways, in order to catabolize certain types of carbohydrates, such as the oligosaccharides, which are present in human breast milk but are not digestible by their host (Bottacini et al., 2014). In one particular clinical study (Braga et al., 2011) was observed that probiotics like B. breve in combination with L. casei are able to prevent the necrotizing enterocolitis because of their positive effects on intestinal motor function, modulation of inflammatory response, and mucosal barrier function. It was also demonstrated that B. infantis administered together with L. casei have improved the intestinal motility in the preterm infants with a very low birth weight associated with the occurrence of necrotizing enterocolitis as a primary outcome (Braga et al., 2011).

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Clinical trials performed on infants accusing acute diarrhea episodes, confirm the fact that a fermented formula for infants using B. breve c50 and Streptococcus thermophillus 0.65, have the ability to alleviate the severity of diarrheal episodes. It is considered that this positive result could be associated with the bifidogenic and the immunostimulatory properties of the fermentation products contained in the fermented product (Aggett et al., 2001; Thibault et al., 2004).

15.5  Microencapsulation of Probiotics An increased number of disorders (i.e., obesity, diabetes, neurological disorders, and cancer) was identified to be enhanced by the imbalance of the microbiota’s structure from the gastrointestinal tract. Many factors like medication usage (antibiotics), differences in diet, Clostridium dificile, and other factors, influence the microbiome composition and lead to the disease induction and its progression (Anselmo et al., 2016). There are some biological impediments encountered when the probiotic strains are delivered through the oral pathway, thereby new ways of introducing the probiotic species into the microbiota structure present huge interest in the research field (Anselmo et al., 2016; Borșa et al., 2015). The novel probiotic-introducing technologies need to confront the oral delivery impediments, like: the acidic pH at the stomach level, the presence of the bile salts which are able to degrade probiotics, and the gastrointestinal transit which limits the adhesion and the growth of probiotics on the intestine surface. Technologies like nanoparticles, enteric coatings, pills, and polymer gels, are specifically designed to prevent the chemical and physical degradation of the interest compounds by the action of acids or enzymes, and to facilitate the mucoadhesion on the gut surface, in order to ensure the interest compounds absorption and its controlled release (Călinoiu et  al., 2016). The delivery of live probiotics to the human microbiome encounters specific challenges due to their large size and their viability and growth-­specific regimes (Pop et al., 2016). For a better protection of probiotics against the gastrointestinal damages, some methods for the probiotics encapsulation (layer-bylayer encapsulation; encapsulation in biopolymeric system; spray drying and freeze drying; and extrusion and emulsion) (Anselmo et al., 2016; Arslan-Tontul and Erbas, 2017; Ashwar et  al., 2018; Huq et  al., 2013; Vodnar et al., 2015) in order to facilitate the passing, the adhesion, and the growth of the bacterial strains to the intestinal surfaces were developed. The encapsulation process presume the wrapping of the interest compound (i.e., live probiotic strains, bioactive ­compounds,

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and drugs) with a special film in order to ensure its viability and its release into the targeted areas of the gastrointestinal tract (Rokka and Rantamaki, 2010). Also, encapsulation represents a process by which a certain material (solid, liquid, or gas) or an amalgam of materials is coated with, or entrapped within another material or system (Huq et al., 2013; Arslan-Tontul and Erbas, 2017). Several technologies of microencapsulation are further described: a. Emulsion—a microencapsulation method with a great potential for the industrial production of the microspheres in a much shorter time. For the emulsification method, aqueous solutions of polysaccharides are scattered in oil phase in order to form a water/oil emulsion, and after that a solution of CaCl2 is added with an uninterrupted stirring for both the emulsification and the encapsulation of probiotics (Ashwar et  al., 2018; Mokarram et  al., 2009). L. plantarum and L. casei can be successfully encapsulated through the emulsion method, when RS-4 (phosphorylated starch type 4) is used. These probiotics are well protected against the gastrointestinal conditions showing a good survival ability on adverse heat treatments and on long-time storage (2 month) (Ashwar et al., 2018). b. Layer-by-layer encapsulation—this novel technology improves the probiotic in  vivo delivery by directly addressing chemical, physical, and probiotic-specific microencapsulation challenges. The most used polymers with great mucoadhesive properties for oral delivery usage are the cationic polysaccharide chitosan and the anionic polysaccharide alginate. These polymers are sequentially layered on the investigated probiotic strains through electrostatic interactions, up to three doubled layers. From the experimental trials when Bacillus coagulans was encapsulated via this technology is outlined that the layer-by-layer encapsulation presents significant advantages considering the survival rate against acids and bile damages, the mucoadhesion, and the direct growth on the gut surfaces. More than that, layer-by-layer microencapsulation could be used also for delivering any valuable and charged compounds, like polyelectrolytes, proteins, or polysaccharides in combination with any other probiotic strains (Anselmo et al., 2016). c. Encapsulation using a bio-polymeric system—is a method to encapsulate probiotics for the food applications. Using this method is expected that the shelf life of probiotics at room temperature in various matrices will be much more increased, and also the heat and stress resistance, and acid tolerance will be even more effective. Mixtures of biopolymers in different concentrations (alginate, chitosan, gelatin, whey protein isolate, cyclodextrin, maltodextrin, xanthan gum, gelatin, and starch) might have the best potential for the encapsulation of probiotics (Ashwar et al., 2018; Huq et al., 2013; Pop et al., 2015).

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The mucilage and the soluble proteins extracted from chia seed and flaxseed in a mixture with maltodextrin are considered potential agents for the encapsulation of probiotics, conferring them a high survival rate and a good viability after spray drying and long-term storage. Also, the mucilage-maltodextrin-soluble proteins mixture ensures the resistance of the probiotics to the simulated bile solution and to gastric juice (Bustamante et al., 2017). Probiotic strains like L. plantarum ATCC 8014 together with the B. infantis ATCC 15679 present high viability (>9 Log10 CFU/g) when they are encapsulated with a mixed material of mucilage-maltodextrin-soluble proteins incorporated in instant juice powder after 45 days of refrigerated storage (Bustamante et al., 2017). Encapsulation with a mixed material reduces the particle size of the probiotic powders thereby offering supplementary functional benefits. d. Spray drying, freeze drying, and spray chilling. Spray drying method helps to produce smaller capsules than by employing other methods. As a side effect, spray drying could produce irreversible damages to the functional entirety of the probiotic cell’s proteins and membranes leading to its inactivation, when high temperatures and pressure (over 130°C, 0.3 bar) are reached. Freeze drying instead might negatively affect the probiotics due to the very low freezing temperatures. Anyhow, a major challenge of spray and freeze drying is to ensure a high stability and viability for probiotics (Arslan-Tontul and Erbas, 2017). The spray chilling method is quite similar with the spray drying, but the significant difference between them is represented by the atomization of the wall materials to the cool chamber. The spray chilling technique mechanism assume the injection of melted matrix to the cold chamber and then a rapidly solidification of the droplets when they come in contact with the cold air flux. The droplets are transformed into microcapsules containing probiotics. So, the important separating factor of the spray chilling from other microencapsulation methods (spray drying and freeze drying) consists in the wall of the used materials (Arslan-Tontul and Erbas, 2017).

15.6  Probiotic Bacteria Viability Under Different Conditions 15.6.1  Processing, Storage, and Food Composition To provide health benefits to the host, it is very important for the probiotic foods to maintain a number of viable and active bacterial cells (over 7 log CFU/g) during their shelf life. Unfortunately, there are several harsh conditions that have a negative impact on the viability

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of probiotic bacteria, such as oxidative stress, storage temperature, humidity, acidity, osmotic pressure, light, etc. Consequently, a major challenge for the food industry is to stabilize the viability of bacteria during production and storage, so the encapsulation process is highly studied in this purpose. There are some techniques used for encapsulation (extrusion, emulsification, spray drying, and fluid bed drying) (Sanchez et al., 2017), the most common being spray drying, because is the cheapest, fastest, and easiest process that can be achieved on industrial scale (Lourenço et al., 2017). As mentioned above, there are a lot of harmful environmental factors which decrease the viability of probiotic bacteria during the biomass production, conservancy, food addition, food storage, and gastrointestinal transit, thus minimizing their health beneficial effect (Burns et al., 2014). These factors can be divided into three categories: food chemical composition (pH, oxygen, salt and sugar content, water activity, chemicals like hydrogen peroxide, artificial flavoring, and heavy metals) (Călinoiu et al., 2016; Murtaza et al., 2017), processing parameters (temperature and exposure time), and microbial parameters (strains of probiotics, growth phase at harvesting, and medium pH). Survival of probiotic cells is limited by a low pH because they have to use their own energy [adenosine triphosphate (ATP)] to maintain intracellular pH. Due to this energy loss, bacteria cannot effectively metabolize nutrients in the environment and gradually begin to die. In addition, the viability of bacteria is constrained by oxygen toxicity, their oxygen exposure causes the formation of toxic metabolites in cells leading to cell death by oxidative damage (Călinoiu et  al., 2016). According to the literature, culture conditions during the fermentation process as well as harvest time can affect the physiological properties of the cells in a way that can contribute to their resistance and stability in the food matrix (Burns et al., 2014). Vinderola et al. revealed in a study conducted in 2012 on Bifidobacterium animalis subspecies lactis INL1 that freeze-dried cells of this species were more resistant to a low pH when grown at pH 5 than when grown with pH 6 (Vinderola et al., 2012). Also, cells harvested in the stationary growth phase are generally more resistant to stressful conditions than those harvested in the exponential phase (Burns et al., 2014). A high survival rate of bacteria can be ensured by their microencapsulation in polymer matrix. The potential of this technique that protects cells from external factors during processing and storage or resistance in gastrointestinal transit has been proven by a vast number of publications (Cook et al., 2012; Huang et al., 2017). An important role in the composition of the food is played by the matrix that will be supplemented with probiotic cells, since the viability of the added bacterium depends on many factors such as the

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acidity of the product, food ingredients and additives, protein, fat, etc. (Profir et al., 2015; Sarao and Arora, 2017). In order to obtain a survival rate of probiotic cells, about 1 × 106–1 × 107, under unfavorable conditions encountered in food matrices, an encapsulation method is required. The most used bacteria in the probiotic preparations are lactic acid bacteria, which include the genera Lactobacillus and Bifidabacterium. Lactobacillus species are naturally found in traditional fermented foods, so they adapt more easily to a lower pH and food substrate like milk (Freire et al., 2017). Probiotic products today are found in supermarkets generally as fermentative dairy products, since the process of juice supplementation is more complicated than the process of dairy products supplementation. Due to the fact that fermented dairy products are not suitable for lactose intolerant consumers, there is an increasing desire to replace that with new food matrices such as fruit and vegetable beverages. These new matrices are valuable as a substrate for probiotics because they already contain beneficial nutrients such as minerals, vitamins, antioxidants, and dietary fiber, and these functional foods can also be consumed by people with lactose intolerance (Profir et al., 2015). The interest for incorporating probiotics into other food matrices, not only in dairy products are increasing, since they are safe for the people who are lactose intolerant (Sarao and Arora, 2017). Bacterial robustness is a key feature to consider when choosing a species for a specific beverage formula. It is proved that in general Lactobacillus are more robust than Bifidobacterium Acidification processes applied to functional beverages limit the selection of strains (Gawkowski and Chikindas, 2013). Several studies have tested the resistance of a number of species of Lactobacillus and Bifidobacterium to acidity by adding them to different juices. In an research conducted by Ravinder et  al. (2012), it was observed that two strains of Latobacillus, namely L. plantarum and L. acidophillus, are able to survive in fermented fruit juices at low pH and high acidity. Besides the fact that these two strains survive in the juice, they are capable to use the fruit juices for their cells synthesis, which is showed by the decrease in sugar and the pH of the fruit and the increase in acidity. It turned out that L. acidophilus consume sugar faster than L. plantar, the decrease in sugar content and pH being faster in the first 24 h and becoming slower over the next 48 h (Ravinder et al., 2012). Watermelon juice has proven to be an effective food matrix for four Lactobacillus species, namely L. casei, L. acidophilus, Lactobacillus fermentum, and L. plantarum. This juice was pasteurized for 30 min at 63°C and was inoculated with a 24-h culture. After an incubation

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of 48 h to 37 degrees, all Lactobacillus species were able to grow in watermelon juice and reach the density of 108 CFU/mL (Sarao and Arora, 2017). A suitable raw material for probiotic beverages has been proven to be tomato juice. Four lactic acid bacteria L. acidophilus LA39, L. plantarum C3, L. casei A4, and Lactobacillus delbrueckii D7 were inoculated in tomato juice, and after 4 weeks of storage at 4°C the number of viable cells ranged from 106 to 108 CFU/mL (Sarao and Arora, 2017). Cashewapple juice has proven to be a good carrier of probiotic bacteria L. casei and can thus be considered as a healthy alternative functional food supplemented with probiotics (Sarao and Arora, 2017). Sheehan et al. (2007) demonstrated that the B. animalis ssp. lactis Bb-12, L. rhamnosus GG, and Lactobacillus paracasei NFBC43338 species remained viable for 12 weeks to over 107 CFU/mL in orange juice and up to 106 CFU/mL in pineapple juice. Although the pH was adjusted to 2.5–3.5, the cranberry juice was found to be lethal within 4 days for viable these probiotic bacteria, the cells were drop under critical value of 106 CFU/mL. The low survival rate in cranberry juice can be attributed to the high benzoic acid content, which in some varieties, is of about 34 mg/L (Sheehan et al. 2007). Survival of probiotic bacteria in fruit juices can be improved by microencapsulation technique in chitosan-coated alginate beads, but consumers’ opinion needs to be considered. These encapsulated probiotics may affect the sensory attributes of the product due to the dimensions of the beads (Sarao and Arora, 2017). Krasaekoopt and Kitsawad conducted in 2010 a study in Thailand that analyzed consumer opinion and sensory evaluation of orange juice and grapefruit juice improved with probiotics. The study found that although the sensory quality of the product was affected by the addition of probiotics, most consumers accepted orange and grapefruit juices enhanced with probiotics beads (82.3% and 84.3%, respectively) (Krasaekoopt and Kitsawad, 2010). In a study conducted by Ding and Shah (2008), the survival rate of eight different species of probiotic bacteria for both free and microencapsulated cells in apple and orange juice has been investigated. The inoculated species were: L. rhamnosus, B. longum, Lactobacillus salivarius, L. plantarum, L. acidophilus, L. paracasei, B. lactis-type Bi-04, and B. lactis-type Bi-07, and their viability tested weekly for 6 weeks. The study found that free bacteria survived in juices only 5 weeks, while encapsulated bacteria were viable during the 6 weeks of storage (Ding and Shah, 2008). As outlined above in line with literature, fruits and vegetables can be used as a food substrate for probiotic bacteria (Vasudha and Mishra, 2013).

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In a previous study, was reported that some cereal ingredients such as lentil and soy flour have increased lactobacilli fractions from the yogurt (Zarea et al. 2012). Over a 10-day storage period, an increase in the viability of Bifidobacterium bifidum bacteria (7.10 log CFU/g) in supplemented yogurt with 2% dates of syrup was observed compared to simple ­yogurt (6.81 log CFU/g), due to a high nutritional value in vitamins, dietary fiber, amino acids, proteins, and carbohydrates (Calinoiu et al., 2016). Chocolate has proven to be a good medium for the growth of probiotic bacteria, but the highest number of probiotic cells could remain in dark chocolate. An explanation might be given by the fact that black chocolate contains a higher amount of cocoa and antioxidant compounds than milk chocolate and white chocolate (Kemsawasd et al., 2016). A study conducted by Dos Santos Cruxen et al. (2017), it has been showed that ice cream, a product considered to be poor in bioactive compounds, can become a functional food by adding fruit that containing bioactive compounds, such Butia odorata, and probiotic bacteria (Dos Santos Cruxen et al., 2017). The bacteria used in this study was B. lactis, showed no significant reduction in concentration within the first 24 h of freezing, remaining viable with a number exceeding 10 over a 90-day period at 18°C (Dos Santos Cruxen et al., 2017).

15.6.2  Freezing, Tawing, and Refrigeration An important role in probiotic cells stability also has its storage temperature, and it was previously reported that encapsulation may have a positive effect on cell viability, especially when exposed to freezing temperatures. Freezing and thawing can affect the integrity and morphology of bacteria, and these negative effects may compromise the protective function of these particles on the viability of probiotics when passing gastrointestinal tract (Sousa et al., 2015). The temperature of −20°C was shown to have a negative effect on the stability of probiotics (L. casei 01, L. paracasei L26, L. acidophilus KI, and B. animalis BB-12) microencapsulated in alginate. After a 60-day storage, some physical changes were observed, such as, spherical shape deformation, increase in microparticle size, therefore, encapsulation in alginate had no protective role during the storage of probiotic cells at the freezing temperatures (Sousa et al., 2015). As mentioned above, an important effect in maintaining the viability of bacteria is its storage temperature and it is known that probiotic yogurts should be stored at the refrigeration temperature. Mortazavian et al. (2007) carried out a study in which it tested the effect of the refrigerated storage temperature on the viability of

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­ robiotics bacteria (L. acidophilus, B. animalis subsp. lactis BB-12) p inoculated in yogurt. The results obtained show that after 20 days of storage, the highest viability for L. acidophilus was obtained when yogurt was stored at 2°C, while for B. lactis the highest viability was obtained when yogurt was kept at 8°C (Mortazavian et al., 2007). Survival of probiotic bacteria, L. casei 01 and L. acidophilus TISTR 450 at low storage temperatures has been investigated in commercial fruit juices such as grapefruit juice, apple, apple, red, and mandarin. Probiotic cells both in the free state and microencapsulated in chitosan-coated alginate beads were added to the five types of juice mentioned above, the products were stored for 4 weeks at 4°C. After the 4 weeks of storage, it was observed that the encapsulated probiotic bacteria had a higher survival rate than the free ones, in addition no significant difference was observed between the types of juice analyzed (Sarao and Arora, 2017). Storage at low temperatures has been shown to be effective in maintaining a survival rate for probiotic bacteria during the storage period (Dianawati et al., 2016).

15.7  Agro-Industrial Wastes and Their Health Potential The market of functional beverages is continuously increasing so the key research priority in this field is to develop new products rich in bioactive compounds targeting lifestyle diseases. Agro-industrial waste is one of the future sector, which must be explored for their bioactive potential. Lately, there is a major issue with respect to the use of agro-industrial waste, industries needing a confirmation of their bioactive potential after thermal processes. This kind of waste is usually produced during the manufacturing processes of the raw materials. Fruit and vegetable juice and pulp industries are generating the highest amount of agro-industrial waste (Dulf et al., 2016, 2017). Waste products deriving from fruits and vegetable processing industries could be even more valorized mainly for two important reasons; (a) due to their low price and their considerable existing amounts and (b) due to their valuable bioactive potential (Vodnar et  al., 2017). Therefore, the following wastes, namely apple peels, carrot pulp, white and red grape peels, and red beet peels and pulp, are in focus being massively discarded; however, this waste is rich in bioactive compounds and can thus be improved and incorporated into food supplements (European Commission Final Report, 2010). According to the experimental study of Vodnar et al. (2017), the demonstrated presence of fatty acids and phenolic compounds in abovementioned agro-­industrial waste, after exposure to a ther-

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mal treatment (10 min, 80°C), enables fruits and vegetable leftovers to become very valuable for the food industry while providing an extra source of income.

15.7.1  Apple Peel Waste Apple peel waste represents one of the major discarded waste within the food industry. The real problem is that this waste has still an impressive bioactive potential, which is not fully exploited, even though many studies reported its capacity to prevent lipid oxidation, to manifest cardioprotective and anticancer properties (Knekt et  al., 2002). According to Boyer and Liu (2004), the responsible bioactive compounds for the reported health-related benefits are the flavonols, the anthocyanins, the flavan-3-ols, the phenolic acids, and the dihydrochalcones, which are present in a huge amount in the apple peel. The results of this study were validated by recent experimental research, which demonstrated that around 80% of the polyphenols are found in the apple peel (Leccese et al., 2009), having a fivefold to sixfold higher antioxidant capacity when compared to apple flesh, also due to unique flavonoids, like quercetin glycosides (Calinoiu et al., 2017).

15.7.2  Carrot Peel and Pulp Waste The carrot peel and pulp waste is another massive generated agrofood waste, which drew attention due to its high amounts of phenolic compounds and dietary fiber. All these bioactive compounds offer particular physical properties to the carrot. For instance, the aroma with specific bitterness, as well as the particular color of carrots are given by anthocyanins and carotenoids (Gonçalves et al., 2010). According to the experimental study of Arscott and Tanumihardjo (2010), carrot waste presents several beneficial health-related properties, like antioxidant potential due to phenolic acids, antiinflammatory effects, and lipid oxidation inhibition due to the anthocyanins presence in cardiovascular disease.

15.7.3  Red and White Grape Peel Waste The next agro-industrial waste in focus is the grape peel waste, both red and white species, due to the huge amount generated during the winemaking process. It is already known the high percentage of grapes, which are used for this purpose, namely 80%, therefore, waste disposal represents a serious issue and practical alternatives for its use and valorization is mandatory. Not to mention its bioactive potential which can be lost due to the strong presence of phenolic compounds (Lafka et  al., 2007), which act as inhibitors of human low-density

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lipoprotein oxidation and have many health-related benefits (Folts, 2002). The strong report among the antioxidant capacity and the total phenolic compounds present was again underlined and demonstrated in the study of Vodnar et al. (2017).

15.7.4  Red Beet Peel and Pulp Waste The red beets are highly appreciated for the betalains content and are considered as important antioxidant vegetables. It is mandatory to underline that around 50% of total phenolics, which are responsible for the antioxidant effect, are present in the peel (Kujala et al., 2000). The red beet peel and its pulp generated by the juice industry represent around 15%–30% from the raw material being considered waste, therefore, discarded as animal feed. Betalains are composed of two types of pigments, namely the red betacyanins and the yellow betaxanthins, which are responsible for the specific color of the beet (Vodnar et  al., 2017). The phenolic compounds present in the peel are as follow: l-tryptophan, p-coumaric and ferulic acids, and cyclodopa glucoside derivatives. Considering its bioactive potential and the health-related benefits, the red beet by-products must be further valorized as functional ingredients or incorporated in different food products/supplements. Also, the juice industry should become aware of their potential and act vigorously by implementing the several waste practical applications demonstrated in the scientific literature: resistance to different thermal processing (Vodnar et al., 2017); atomization under powder form as functional ingredients (Santos et al., 2017).

15.8  Probiotic Nondiary Beverages Based on Targeted Agro-Industrial Wastes The development of new probiotic nondiary functional beverages, with an inflated demand from the lactose-intolerant and vegetarian population is in vogue. Therefore, an emerging trend with respect of designing a probiotic beverage is fortification with selected strains (L. casei, L. plantarum, B. infantis, and B. breve) having a positive impact on gastrointestinal health, targeting the IBS. Clinical studies were done in order to test the efficacy of these probiotic strains. Moreover, the valorizing of agro-industrial wastes by their incorporation into these types of beverages, due to their high-valuable bioactive potential, which may have health-promoting characteristics, and their disposal may harm the environment by contributing to pollution, comprises the innovative idea lying behind the future of

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beverages science. The presence of bioactive molecules, such as fatty acids and phenolic compounds, in agro-industrial waste makes fruit and vegetable leftovers more valuable for the food industry. Fruits and vegetables contain bioactive compounds that impart health benefits beyond basic nutrition (Oomah and Mazza, 2000). These wastes are rich in bioactive compounds and can thus be improved and incorporated into food supplements (European Commission Final Report, 2010). Thus, the design of probiotic functional beverages, valorizing the agro-industrial wastes with microencapsulated bacteria (L. casei, L. plantarum, B. infantis, and B. breve), using smart bottles for product stability and controlled release of the microcapsules is the innovative approach for the emerging market of functional beverages. In this way, based on Vodnar et al. (2017) experimental research study, where the bioactive potential of several agro-industrial waste, after exposure to the thermal treatment, was validated, even enhanced by the process, and continuing with his demonstrated research studies on probiotic microencapsulation (Vodnar et al., 2015; Vodnar and Socaciu, 2014), all packed in a smart packaging bottle with a controlled cap release, was designed and is illustrated in Fig. 15.1.

Fig. 15.1  Design of a nondairy probiotic functional beverage based on agroindustrial waste.

512  Chapter 15  A NEW GENERATION OF PROBIOTIC FUNCTIONAL BEVERAGES

15.8.1  Bioactive Potential of Waste as Fresh and Thermally Processed Matrices In order to underline the bioactive potential of targeted agro-­ industrial waste, for its further valorization, the latest experimental research studies of Vodnar et al. (2017) and Calinoiu et al. (2017) will be the starting point. In Vodnar et al. (2017) study, the total phenolic content, the total flavonoids content, and the antioxidant activity of the waste extracts, both fresh and thermally processed (10 min, 80°C), were examined, as illustrated in Fig. 15.2. For some of the waste extracts, the thermal process enhanced their bioactive potential, while for others was opposite. With respect to the total phenolic content (Fig. 15.2), the thermally processed samples of apple and red grapes had higher total phenolic content, red grape waste had the highest (1990 ± 52.9 mg GAE/100 g dry weight) while for the carrot and red beet waste extracts the thermal process decreased the phenolic content. The increase of phenolic content was possible due to the intracellular contents extraction enhanced by the thermal treatment, where Wang et al. (2014) underlined the reason behind it, namely the hydrolysis of polysaccharides. According to the literature, the decrease [in closure time (CT) and bleeding time (BT)] could be attributed to the partial degradation of lignin, responsible for the release of phenolic acids, or to the thermal degradation of the phenolic acids (Morales de la Pena et al., 2011).

Fig. 15.2  Smart bottle for functional probiotic beverages.

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Considering further the total flavonoids content (Fig. 15.2) according to Vodnar et al. (2017) study, the thermal process enhanced the red grape waste content (up to 1050 ± 62.1 mg), while for apple and white grape waste no significant differences among fresh and thermally processed was observed. In accordance with the increased flavonoids content after the process exposure is also the study of Medina-Meza and Barbosa-Cánovas (2015), where the exposure to an electric field increased by 96% the flavonoid content of grape peel. The antioxidant capacity of targeted waste (Fig. 15.2) was significantly enhanced by the thermal process, for instance red grape waste showed a 59% increase in radical inhibition capacity. In Table 15.5, from Vodnar et al. (2017) experimental study, the identification of the phenolic compounds in the targeted waste extracts was done in order to validate their bioactive potential. In almost all phenolic cases, the fresh and thermally processed samples had significantly different values. Therefore, a significant increase in malvidin glucoside (13.68%) was registered after thermal processing for red grape waste; another major example was reported for caffeic acid 4-O-glucoside from apple waste, with a 27% [from 2.492 (mg%) to 3.165 (mg%)] increase after the thermal treatment. With respect to red beet waste extract, all the reported betanidin compounds increased being enhanced by the thermal treatment exposure. The same result was reported by Harivaindaran et al. (2008) for dragon fruit where the exposure at 100°C for 5 min increased the yield of betacyanin content. The fatty acid profile of each targeted waste sample is shown in Table 15.6, results reported by Vodnar et al. (2017). The thermal treatment increased the fatty acids content in almost all the cases, probably due to cell walls breaking and release of fatty acids. Also, the thermally processed apple waste had the highest content of fatty acids, and the red and white grape waste significantly increased their fatty acids composition/content after exposure. It is known that linoleic acid is very sensitive to the heat, therefore, after exposure to the thermal process, it significantly decreased. In the study of Dulf et al. (2016), it was demonstrated that exposure to certain fermentations increased significantly the fatty acids composition and content. Therefore, different treatments were validated as beneficial for exploiting and enhancing the bioactive potential of several wastes. These results offer sustainable solution for the utilization of food industry biowaste that has bioactive potential after thermal treatment (10 min at 80°C). The apple peels, carrot pulp, red and white grape peels, and red beet peels and pulp can be exploited for their bioactive compounds, whose bioavailability increased and can be added in food formulations, like functional beverages, as health-promoting products.

Table 15.5  Identification of the Phenolic Compounds (mg/100 g DW) in the Waste Extracts Via HPLC-DAD-ESI-MS Method (License Number: 4187141441540) Phenolic Compounds Anthocyanins

Cinnamic acid

Petunidin 3-O(6′-p-coumaroyl)glucoside) Malvidin 3-O-(6′p-coumaroyl)glucoside Cyanidin 3-O-arabinoside Peonidin 3-O-glucoside Malvidin 3-O-glucoside Caffeic acid Caffeic acid-4-O-glucoside 5-Caffeoylquinic acid 3-Caffeoylquinic acid 3,4-Dicaffeoylquinic acid

[M+H]+ Ion Fragments

Samples (mg/%DW) AF

AT

CF

CT

WGF

WGT

RGF

RGT a

BF b

625, 317

2.454

2.356

639, 331

7.358b

8.365a

419, 287

7.745 b

7.625 a

463, 301

2.984a

1.892b

493, 331

13.015b

13.958a

181, 163 343, 181,163

2.492

b

3.165

355, 181, 163 355, 181,163 515, 355

1.27b

2.138b

a

14.146a

4.265b

2.457a

0.263b

2.65a

2.56a

BT

Dihydrochalcones

Flavan-3-ols

Flavonol

Betacyanins Betanidin

Phloridzin (phloretin 2′-O-glucoside) Phloretin 2′-Oxylosyl-glucoside Epicatechin

437, 275 569, 437, 275

5.714a 1.125a

4.652b 0.958b

291

2.368

a

a

2.262

2.261 b

2.265a

Catechin 3-O-glucose Rutin (Quercetin 3-O-rutinoside) Quercitrin (Quercetin 3-O-rhamnoside) Quercetin 3,4′-O-diglucoside

453, 291

Betanidin Isobetanidin Betanidin-5-O-βglucoside Isobetanidin-5-O-βglucoside

389,345 389, 345 551, 389

3.866b 2.353b 7.006b

3.952a 2.456a 7.100a

551, 389

10.061a

10.074a

611, 303

4.318 a

4.316a

449, 303

2.12b

2.46a

627, 465,303

0.903b

1.362a

Values (mean ± SD, n = 3) in the same row followed by different superscript letters (a, b) indicate significant differences (P