Microbial Biotechnology in Food Processing and Health: Advances, Challenges, and Potential 1st Edition Deepak Kumar Verma (Editor)

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MICROBIAL BIOTECHNOLOGY
IN FOOD PROCESSING
AND HEALTH
Advances, Challenges, and Potential
MICROBIAL BIOTECHNOLOGY
IN FOOD PROCESSING
AND HEALTH
Advances, Challenges, and Potential
Edited by
Deepak Kumar Verma
Ami R. Patel, PhD
Sudhanshu Billoria, PhD
Geetanjali Kaushik, PhD
Maninder Kaur, PhD
First edition published 2023
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Library and Archives Canada Cataloguing in Publication

Title: Microbial biotechnology in food processing and health : advances, challenges, and potential / edited by
Deepak Kumar Verma, Ami R. Patel, PhD, Sudhanshu Billoria, PhD, Geetanjali Kaushik, PhD, Maninder Kaur, PhD.
Names: Verma, Deepak Kumar, 1986- editor. | Patel, Ami R., editor. | Billoria, Sudhanshu, editor. | Kaushik, Geetanjali, editor. |
Kaur, Maninder, editor.
Description: First edition. | Includes bibliographical references and index.
Identifiers: Canadiana (print) 20220204993 | Canadiana (ebook) 20220205051 | ISBN 9781774637289 (hardcover) |
ISBN 9781774637432 (softcover) | ISBN 9781003277415 (ebook)
Subjects: LCSH: Probiotics. | LCSH: Microbial metabolites. | LCSH: Functional foods. | LCSH: Microbial biotechnology. |
LCSH: Food industry and trade.
Classification: LCC RM666.P835 M53 2023 | DDC 615.3/29—dc23
Library of Congress Cataloging-in-Publication Data
CIP data on file with US Library of Congress
ISBN: 978-1-77463-728-9 (hbk)

ISBN: 978-1-77463-743-2 (pbk)
ISBN: 978-1-00327-741-5 (ebk)
Dedication
This book is dedicated to the eminent Professor Hari Niwas Mishra, of the
AgFE Department, IIT Kharagpur, West Bengal, India.
Dr. Mishra has made wonderful and remarkable contributions to the growth
of knowledge and research in the field of food science and technology with
his vast experience in the administration and management of industrial-
scale collaborative research projects ranging from new processes, product
development, and also creation of pilot-scale facilities. He focuses on devel
oping, enhancing, and commercializing food technologies by collaborating
with industry to promote product diversification and enhancement, increase
market growth, and aid competitiveness.
About the Editors
Deepak Kumar Verma, PhD

Department of Agricultural and Food
Engineering, Indian Institute of Technology
Kharagpur, West Bengal, India
Deepak Kumar Verma, PhD, is an agricultural

science professional and has received PhD
degree with specialization in Food Processing
Engineering from Agricultural and Food
Engineering Department, Indian Institute
of Technology Kharagpur (West Bengal),
India. In 2012, he received a DST-INSPIRE Fellowship for PhD study by
the Department of Science and Technology (DST), Ministry of Science
and Technology, Government of India. Mr. Verma is currently assigned
for research on “Aroma Volatile and Flavoring Compounds from Indian
Rice Cultivars” whereas during his master’s degree, his research was on
“Physicochemical and Cooking Characteristics of Azad Basmati (CSAR
839-3): A Newly Evolved Variety of Basmati Rice (Oryza sativa L.).” He
earned his BSc degree in agricultural science in 2009 from the Faculty of
Agriculture, Gorakhpur University, Gorakhpur, and MSc (Agriculture) in
Agricultural Biochemistry in 2011 with the first rank and also received a
department topper award from the Department of Agricultural Biochemistry,
Chandra Shekhar Azad University of Agricultural and Technology, Kanpur,
India. Apart from his area of specialization in plant biochemistry, he has
also built-up a sound background in plant physiology, microbiology, plant
pathology, genetics, and plant breeding, plant biotechnology, and genetic
engineering, seed science and technology, food science and technology, etc.
In addition, he is a member of various professional bodies, and his activities
and accomplishments include conferences, seminars, workshops, training,
and also the publication of research articles, books, and book chapters.
viii About the Editors
Ami R. Patel, PhD

Assistant Professor, Division of Dairy and
Food Microbiology, Mansinhbhai Institute of
Dairy & Food Technology, Dudhsagar Dairy
Campus, Gujarat, India
Ami R. Patel, PhD, is an Assistant Professor in

the Division of Dairy and Food Microbiology,
Mansinhbhai Institute of Dairy & Food
Technology-MIDFT, Dudhsagar Dairy Campus,
Gujarat, India. He has expertise in specialized
areas that involve isolation, screening, and
characterization of exopolysaccharides from potential probiotic cultures
and employing them for food and health applications. In addition, she is
engaged with teaching undergraduate, postgraduate, and research students
in food microbiology, microbial biotechnology, food biotechnology, food
science and technology, clinical microbiology and immunology, etc. She
has authored peer-reviewed papers and technical articles in international
and national journals as well as book chapters, books, proceedings, and
technical bulletins. She has received awards and honors, including the Vice
Chancellor Gold Medal for her PhD work, selection as a BiovisinNxt11
fellow to attend an international conference at Lyon, France, and an Erasmus
Mundus scholarship from the European Union for three years to visit Lund
University, Sweden, as a guest researcher. She is also serving as an expert
reviewer for several scientific journals. She is a member of several academic
and professional organizations, including the Indian Dairy Association and
the Swedish South Asian Network for Fermented Foods.
Dr. Patel earned her BSc (Microbiology) and her MSc (Microbiology)
degree from Sardar Patel University, Gujarat, and received her doctorate
degree in Dairy Microbiology from the Dairy Department of SMC College
of Dairy Science, Anand Agricultural University, Gujarat, India.
About the Editors ix
Sudhanshu Billoria, PhD

Assistant Professor of Production (Food
Processing), Center for Research, Consultancy
and Publication, Vaikunth Mehta National
Institute of Cooperative Management
(VAMNICOM), Pune, Maharashtra, India
Sudhanshu Billoria, PhD, is an Assistant

Professor of Production (Food Processing)
at the Center for Research, Consultancy and
Publication, Vaikunth Mehta National Institute
of Cooperative Management (VAMNICOM), Pune, Maharashtra, India,
where she teaches various postgraduate-level courses and guides research
projects. Her research interests include postharvest management of fresh
produce, mainly fruits and vegetables, by extending shelf life. She utilizes
her expertise for addressing value-chain and supply chain management
and industrial cooperatives such as dairy, sugar, and fruit and vegetable
cooperatives. She has published many research papers in peer-reviewed
international journals and conference proceedings along with many book
chapters. She is a member of various professional bodies and has attended,
presented papers, and delivered invited lectures at workshops, conferences,
and training programs.
Dr. Billoria earned an MSc degree in Food Science and Technology
from the University of Jammu, Jammu and Kashmir, India, and her PhD
in Food Process Engineering from the Agricultural and Food Engineering
Department, Indian Institute of Technology Kharagpur, wherein her
research work was "Shelf-Life Extension of Light Red Color Tomatoes
(Lycopersicon esculentum cv. Vaishali) with the Help of Active Packaging
and Edible Coating Technologies.” Her master’s project work was entitled
“Standardization and Preparation of Ghee based Butter.”
x About the Editors
Geetanjali Kaushik, PhD

Associate Professor, Department of Civil
Engineering and Head of R&D, Hi Tech
Institute of Technology, Waluj, Maharashtra,
India
Geetanjali Kaushik, PhD, is an Associate

Professor in Environment at Hi Tech Institute of
Technology Waluj Maharashtra. Dr. Geetanjali
Kaushik completed her PhD in Pesticide
residues in Food, from the prestigious Indian
Institute of Technology Delhi with more than 75 International papers and
chapters in Journals and Books of repute. She has participated in International
Conferences held in Germany (won first prize) and USA (received American
Society of Nutrition travel grant). Subsequent to her PhD she has also worked
with University of California, Berkley on an air quality monitoring project for
over 40 locations across Delhi. She has also edited 6 International books. Her
taught assignments include stints as Assistant Professor with the University
of Petroleum and Energy Studies, Dehradun, and Amity University, Noida.
In 2012, Dr. Kaushik received full scholarship from School of Management,
Swansea University UK to undertake MBA. She topped the MBA class and
has worked in UK and Turkey in various positions. She returned to India in
2015 and has worked as an Associate Professor with MGM’s Jawaharlal Nehru
Engineering College, Aurangabad. Currently she is working as an Associate
Professor in Dept of Civil Engineering and Head R&D at Hi Tech Institute
of Technology Waluj Maharashtra. She has prepared Air Action Plans for
cities of Aurangabad, Ulhasnagar, Jalna, Latur, Nashik, Thane, Jalgaon, and
Badlapur which have been approved for funding and further implementation
by CPCB under NCAP (National Clean Air Program). She has also prepared
and submitted the Environment Status Report 2017–2018, 2018–2019 for
Aurangabad city and 2018–2019 for Ulhasnagar city. Dr. Kaushik is also
undertaking Maha YUVA (Maharashtra Youth Understanding Value of Air)
Campaigns and Awards 2019–2020 in cities of Aurangabad, Jalna, and
Latur for school and college students. This project is sponsored by MPCB.
Dr. Geetanjali also organized 11 workshops in Aurangabad city in
collaboration with EPIC (Energy Policy Institute at University of Chicago).
Dr. Kaushik is also the nodal person for implementation of Unnat Bharat
Abhiyan for Hi-Tech Institute of Technology. This is a flagship program of
MHRD being implemented by IIT Delhi.
About the Editors xi
Maninder Kaur, PhD

Associate Professor, Department of Food
Science and Technology, Guru Nanak Dev
University, Amritsar, Punjab, India
Maninder Kaur, PhD, is working as an Associate

Professor in the Department of Food Science
and Technology, Guru Nanak Dev University,
Amritsar, Punjab, India. In 1997, She earned
her BSc (Home Science); in 1999, MSc (Food
Technology) degree and received her doctorate
degree in Food Technology from the Department of Food Science and
Technology, Guru Nanak Dev University, Amritsar, Punjab, India in the
year 2005. Dr. Kaur has her expertise in a specialized area which involves
Characterization of flours, starches, and proteins from different botanical
sources. In addition, she is engaged with teaching UG, PG, and PhD students
at Guru Nanak Dev University, Amritsar, India. She has authored a number
of peers reviewed papers, technical articles, book chapters, in international
and national journals, books, proceedings, and technical bulletins.
Prof. Kaur has received a number of awards and honors, such as: awarded
Post-Doctoral Research Fellowship by Korean Ministry of Education, South
Korea and selected for Young Scientist Award-2010 by Association of Food
Scientists and Technologists (India). She is serving as an expert reviewer for
several scientific journals. She is a member of academic and professional
organizations like Association of Food Scientists and Technologists (AFSTs),
Mysore, India, and Punjab Academy of Sciences (PASs).
Contents
Contributors........................................................................................................... xv
Abbreviations ........................................................................................................ xix
Symbols ............................................................................................................... xxiii
Preface ..................................................................................................................xxv
Part I: Probiotics and Their Metabolites: A Food Industry Perspective .......... 1

1. Action of Probiotic Microorganisms on Mycotoxin
Decontamination in Milk and Milk Products.............................................. 3
Katia Francine Wochner, Tânia Aparecida Becker-Algeri, Eliane Colla,
and Deisy Alessandra Drunkler
2. New Technological Trends in Probiotics Encapsulation for

Their Stability Improvement in Functional Foods and
Gastrointestinal Tract.................................................................................. 21
Majid Nooshkam and Zahra Zareie
3. Bioactive Peptides (BPs) as Functional Foods: Production Process,

Techno-Functional Applications, Health-Promoting Effects,
and Safety Issues .......................................................................................... 51
Afshin Babazadeh, Majid Nooshkam, and Mahnaz Tabibiazar
Part II: Probiotics and Potential Health Benefits ............................................. 79

4. Probiotics in Fruits and Vegetables: Challenges,
Legislation Issues, and Potential Health Benefits...................................... 81
Mamta Thakur, Deepak Kumar Verma, Sudhanshu Billoria, H. W. Deshpande,
Ami R. Patel, and Geetanjali Kaushik
5. Ice Cream as Probiotic Food and Its Potential Benefits in

Human Health ............................................................................................ 129
Tatiana Colombo Pimentel, Michele Rosset, Suellen Jensen Klososki,
Carlos Eduardo Barão, Vanessa Aparecida Marcolino,
Vanessa Guimarães Alves Olher, and Adriano Gomes da Cruz
Part III: Industrial Aspects of Microbial Food Technologies ........................ 165

6. Lactulose: A High Food Value-Added Compound and
Its Industrial Application in Food ............................................................ 167
Majid Nooshkam and Zahra Zareie
xiv Contents
7. Innovation in Technology for Processing of Sheep and

Goat Milk and Dairy Products: An Overview......................................... 199
Antonio José Trujillo and Bibiana Juan
8. Sustainable Use of Food-Grade Microorganisms in

Traditional Fermented Food Production................................................. 237
Kolawole Banwo, Omotade R. Ogunremi, and Adekemi T. Adesulu-Dahunsi
9. Advanced Molecular Tools and Techniques for Assessment of

Microbial Diversity in Fermented Food Products .................................. 263
Damanpreet Kaur, Sushma Gurumayum, Prasad Rasane, Sawinder Kaur,
Jyoti Singh, Navneet Kaur, Kajal Dhawan, and Ashwani Kumar
10. Industrial Prospects of Bacterial Microcompartment Technologies..... 297

Shagun Rastogi and Chiranjit Chowdhury
11. Management of Microbiological Hazards in the

Food Processing Industries ....................................................................... 323
Tejpal Dhewa and Anu Kumar
Index .................................................................................................................... 337

Contributors
Adekemi T. Adesulu-Dahunsi
Department of Microbiology, Landmark University, Omu-Aran, Kwara State, Nigeria,
E-mail: adesuluchemmy@yahoo.com
Afshin Babazadeh
Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Macquarie University,
Sydney, NSW – 2109, Australia, E-mail: babazadeh.afshin@gmail.com
Kolawole Banwo
Department of Microbiology, University of Ibadan, Oyo State, Nigeria, E-mail: kolabanwo@yahoo.com
Carlos Eduardo Barão

Federal Institute of Paraná (IFPR), Paranavaí – 87703-536, Paraná, Brazil,
E-mail: carlos.barao@ifpr.edu.br
Tânia Aparecida Becker-Algeri
Dra. Research in Food Science and Technology, Quality Management Department,
Cooperative Agro-Industrial Lar, Avenida 24 de Outubro, n 59, Industrial Area, Medianeira,
Paraná, CEP – 85884000, Brazil, E-mail: taniabecker86@yahoo.com.br
Sudhanshu Billoria
Center for Research, Consultancy, and Publication, Vaikunth Mehta National Institute of Cooperative
Management, Pune, Maharashtra – 411007, India, E-mail: sudharihant@gmail.com
Chiranjit Chowdhury
Amity Institute of Molecular Medicine and Stem Cell Research, Amity University Uttar Pradesh,
Sector-125, Noida – 201313, Uttar Pradesh, India,
E-mails: cchowdhury@amity.edu; cchowdhury82@gmail.com
Eliane Colla
Dra. Research in Food Science and Technology Post Graduate Program in Food Technology (PPGTA)
Department, Federal Technological University of Paraná (UTFPR), Medianeira, Paraná, Brazil,
E-mail: ecolla@utfpr.edu.br
Adriano Gomes da Cruz

Rio de Janeiro Federal Institute of Education, Science, and Technology (IFRJ),
Professional Master’s in Food Science and Technology of (PGCTA), Maracanã – 20270-021,
Rio de Janeiro, Brazil, E-mail: food@globo.com
H. W. Deshpande
Department of Food and Industrial Microbiology, College of Food Technology,
Vasantrao Naik Marathwada Krishi Vidyapeeth, Parbhani – 431402, Maharashtra, India,
E-mail: hemantd22@gmail.com
Kajal Dhawan
Research Scholar, Department of Food Technology and Nutrition, Lovely Professional University,
Phagwara – 144411, Punjab, India, E-mail: kajaldhawan42@gmail.com
Tejpal Dhewa
Department of Nutrition Biology, Central University of Haryana, Mahendergarh – 123031, Haryana,
India, E-mails: tejpal_dhewa07@rediffmail.com; tejpaldhewa@cuh.ac.in
xvi Contributors
Deisy Alessandra Drunkler

Dra. Research in Food Science and Technology, Post Graduate Program in Food Technology (PPGTA)
Department, Federal Technological University of Paraná (UTFPR), Medianeira, Paraná, Brazil,
E-mail: deisydrunkler@utfpr.edu.br
Sushma Gurumayum
Assistant Professor, Department of Basic Engineering and Applied Sciences, College of Agricultural
Engineering and Post-Harvest Technology, Central Agricultural University, Ranipool – 737135,
Sikkim, India, E-mail: sushmagurumayum@gmail.com
Bibiana Juan
Center for Innovation, Research, and Transfer in Food Technology (CIRTTA), XaRTA, TECNIO,
MALTA Consolider, Department of Animal Science and Food Sciences,
Faculty of Veterinary Medicine, Universitat Autònoma de Barcelona, Bellaterra – 08193, Spain,
Tel.: +34-935811447, Fax: +34-935812006, E-mail: bibiana.juan@uab.es
Damanpreet Kaur
Research Scholar, Department of Food Technology and Nutrition, Lovely Professional University,
Phagwara – 144411, Punjab, India, E-mail: preet.daman9624@gmail.com
Navneet Kaur
School of Engineering Technology and Applied Science, Centennial College, Toronto, Canada,
E-mail: nkaur253@my.centennialcollege.ca
Sawinder Kaur
Department of Food Technology and Nutrition, Lovely Professional University, Phagwara – 144411,
Punjab, India, E-mail: sawi_raman@yahoo.co.in
Geetanjali Kaushik
Department of Civil Engineering, HiTech Institute of Technology, Waluj Aurangabad – 431136,
Maharashtra, India, E-mail: geetanjaliac@gmail.com
Suellen Jensen Klososki

E-mail: suellen.jensen@ifpr.edu.br
Anu Kumar
Department of Biotechnology, Chandigarh University, Gharuan, Mohali – 140301, Punjab, India,
E-mails: anubiotech88cu@gmail.com; anubiotech88@gmail.com
Ashwani Kumar
Assistant Professor, Department of Food Technology and Nutrition, Lovely Professional University,
Phagwara – 144411, Punjab, India, E-mail: ashwanichandel480@gmail.com
Vanessa Aparecida Marcolino

E-mail: vanessa.marcolino@ifpr.edu.br
Majid Nooshkam
Department of Food Science and Technology, Faculty of Agriculture,
Ferdowsi University of Mashhad (FUM), Mashhad, Iran, Tel.: +98-51-38795620,
Mobile: +98-9169448412, Fax: +98-51-38787430, E-mail: Nooshkamma@gmail.com
Omotade R. Ogunremi
Department of Microbiology, Technical University, Ibadan, Oyo State, Nigeria,
E-mail: tadeogunremi@yahoo.com
Contributors xvii
Vanessa Guimarães Alves Olher

E-mail: vanessa.olher@ifpr.edu.br
Ami R. Patel
Assistant Professor, Mansinhbhai Institute of Dairy and Food Technology (MIDFT),
Dudhsagar Dairy Campus, Mehsana – 384002, Gujarat, India,
E-mails: amimicro@gmail.com; ami@midft.com
Tatiana Colombo Pimentel
E-mail: tatiana.pimentel@ifpr.edu.br
Prasad Rasane
Assistant Professor, Department of Food Technology and Nutrition,
Lovely Professional University, Phagwara – 144411, Punjab, India,
E-mails: rasaneprasad@gmail.com; prasadrasane4u@gmail.com
Shagun Rastogi
Amity Institute of Biotechnology, Amity University Uttar Pradesh, Sector-125, Noida – 201313,
Uttar Pradesh, India, E-mail: shagun177@gmail.com
Michele Rosset
Federal Institute of Paraná (IFPR), Colombo – 83403-515, Paraná, Brazil,
E-mail: michele.rosset@ifpr.edu.br
Jyoti Singh
Assistant Professor, Department of Food Technology and Nutrition, Lovely Professional University,
Phagwara – 144411, Punjab, India, E-mail: jyotisingh9377@gmail.com
Mahnaz Tabibiazar
Nutrition Research Center and Department of Food Science and Technology,
Faculty of Nutrition and Food Sciences, Tabriz University of Medical Sciences, Tabriz, Iran,
E-mail: mahnaz_tabibiazar@yahoo.com
Mamta Thakur
Department of Food Engineering and Technology, Sant Longowal Institute of Engineering and
Technology, Longowal – 148106, Punjab, India,
E-mails: thakurmamtafoodtech@gmail.com; mamta.ft@gmail.com
Antonio José Trujillo

Center for Innovation, Research, and Transfer in Food Technology (CIRTTA), XaRTA, TECNIO,
MALTA Consolider, Department of Animal Science and Food Sciences,
Faculty of Veterinary Medicine, Universitat Autònoma de Barcelona, Bellaterra – 08193, Spain,
Tel.: +34.935813292, Fax: +34-935812006, E-mail: Toni.Trujillo@uab.es
Deepak Kumar Verma

Agricultural and Food Engineering Department, Indian Institute of Technology, Kharagpur – 721302,
West Bengal, India, Tel.: +91-3222281673, Mobile: +91-7407170259, Fax: +91-3222282224,
E-mails: deepak.verma@agfe.iitkgp.ernet.in; rajadkv@rediffmail.com
Katia Francine Wochner

Ma. Research in Food Science and Technology, Post Graduate Program in Food Technology (PPGTA)
Department, Federal Technological University of Paraná (UTFPR), Avenida Brasil, 4232,
Bairro Parque Independência, Medianeira, Paraná, CEP – 85884-000,
Brazil, E-mail: katia_kfw29@hotmail.com
xviii Contributors
Zahra Zareie
Department of Food Science and Technology, Gorgan University of Agricultural Sciences and Natural
Resources, Gorgan, Iran, E-mail: Zareie91@gmail.com
Abbreviations
ACE angiotensin converting enzyme

AFB1 aflatoxin B1
AFLP amplified-fragment length polymorphism
AFM1 aflatoxin M1
ARDRA amplified ribosomal DNA restriction analysis
BEN Balkan endemic nephropathy
BLIS bacteriocin like inhibitory substances
BMV bacterial microcompartment vertex
BOD biological oxygen demands
BPs bioactive peptides
CAGR compound annual growth rate
CC codex committees
CCPs casein calcium peptides
CD Crohn’s disease
CE-MS capillary electrophoresis mass spectrometry
CF curd firmness
Cfu colony-forming unit
COD chemical oxygen demands
CODES colon targeted delivery system
CPC consumer protection commission
CPP casein phosphor-peptides
EC European Commission
EFSA European Food Safety Authority
EPS exopolysaccharide
EU European Union
Eut ethanolamine utilization
EutM Eut MCP
FAO Food and Agriculture Organization
FB1 fumonisin B1
FDA Food and Drug Administration
FOS fructo-oligosaccharides
FSSAI Food Safety and Standards Authority of India
FT-IR Fourier transform infrared
GC-MS gas chromatography-mass spectrometry
xx Abbreviations
GFP green fluorescent protein

GI gastrointestinal
GIT gastrointestinal tract
GMPs good manufacturing practices
GOS galactooligosaccharides
GRAS generally recognized as safe
GST glutathione S-transferase
HACCP hazard analysis and critical control point
HHP high hydrostatic pressure
HPH high pressure homogenization
HPMC hydroxyl propyl methyl cellulose
HPP high-pressure processing
HTS high-throughput sequence
HTST high temperature-short time
IAT impinging aerosol technology
IBD inflammable bowel disease
IBS irritable bowel syndrome
Igs immunoglobulins
IMO isomaltooligosaccharide
LAB lactic acid bacteria
LC-MS liquid chromatography-mass spectrometry
LTLT low-temperature long-time
MAP modified atmosphere packaging
MBP maltose-binding protein
MC moisture content
mCOLD-PCR modified CO-amplification at low denaturation tempera
ture PCR
MCP microcompartment
MEA malt extract agar
MLSA multilocus sequence analysis
MLST multilocus sequence typing
MRPs Maillard reaction products
MRS Man Rogosa Sharpe
MS mass-spectrometry
NAFDAC National Agency for Food and Drug Administration and
Control
NFSAP National Food Safety Action Plan
NGS next generation sequencing
NMR nuclear magnetic resonance
Abbreviations xxi
O/W oil-in-water
OTA ochratoxin
OTU operational taxonomic unit
PCA-BCP plate count agar with bromocresol purple
PCR-DGGE polymerase chain reaction-denaturing gradient gel
electrophoresis
Pdu propanediol utilization
PEFs pulsed electric fields
PMA propidium monoazide dye
PR pressure-treated
PVOH polymer of polyvinyl alcohol
qPCR quantitative PCR
RAPD random amplification of polymorphic DNA
RCF rate of curd firming
RCT rennet clotting time
RFLP restriction fragment length polymorphism
SAB Slanetz and Bartley medium
SDF soluble dietary fiber
SERB Science and Engineering Research Board
SHR spontaneously hypertensive rats
SMEs small and medium-scale enterprises
SPI soy protein isolate
SSCP single stranded confirmation polymorphisms
TCA tricarboxylic acid cycle
UC ulcerative colitis
UHPH ultra-high-pressure homogenization
UHT ultra-high temperature
UNL University of Nebraska-Lincoln
USA United States of America
UTI urinary tract infection
VBNC viable but non-culturable
W/O water-in-oil
W/O/W water-in-oil-in-water
WGPH wheat germ protein hydrolysates
WHO World Health Organization
WPC whey protein concentrate
WPI whey protein isolate
WTO World Trade Organization
YGC yeast glucose chloramphenicol
xxii Abbreviations
YPD yeast extract potato dextrose agar

ZEA zearalenone
α-La α-lactalbumin
α-ZOL α-zearalenol
β-Lg β-lactoglobulin
Symbols
% percentage
˚C degree Celsius
µm micrometer
h hour
Kg kilogram
mg milligram
ml milliliter
pH power of hydrogen
Tg glass transition temperature
w/v weight/volume
Preface
The use of foods to improve health and the state of well-being is progres
sively more accepted by both science and society apart from the primary role
of food to provide nutrients for performing normal physiological functions.
Probiotics (health beneficiary bacteria) or their metabolites such as bacte
riocins, bioactive peptides (BPs), etc., and prebiotics (usually functional
non-digestible oligosaccharides) are considered as a key biotechnological
field with tremendous potential for innovation. Additionally, knowledge of
microbial communities is essential for shaping the final characteristics of
diverse food products. Consequently, this book describes the understanding
and dissemination of knowledge surrounding probiotics and/or their
metabolites, prebiotics in various food products, and human health, along
with recent trends in microbial fermentation and innovative technologies in
academic researches.
This book Microbial Biotechnology in Food Processing and Health:
Advances, Challenges, and Potential, is divided into three main parts: Part
I: Probiotics and Their Metabolites: Food Industry Perspective; Part II:
Probiotics and Potential Health Benefits; and Part III: Industrial Aspects
of Microbial Food Technologies. Probiotic cultures are employed for the
manufacturing of dairy as well as non-dairy-based fermented or non-
fermented food products. Recently, screening and designing functional
starters with technological aid advantages are fascinating areas of investiga
tion in the food sector. In this context, chapters compiled in Part I discuss
the types and roles of beneficial microbes and/or their metabolites in food
products such as enhancing food safety by decontaminating or neutralizing
toxic components like mycotoxins associated with foods or improving their
stability in functional foods. The health-beneficial microorganisms known
as probiotics have been proven for many immunomodulatory and immuno
stimulatory effects in a number of chronic infections and diseases.
Hence, in Part II of the book, recent breakthroughs in the development
of novel probiotics-incorporated dairy and non-dairy (fruits and vegetables)
based food products, challenges associated with their commercialization,
and studied health benefits have been covered. Innovative approaches are
employed in food science for various means; hence in Part III, emerging
xxvi Preface
technologies dealing with assessing microbial diversities or management of

microbiological hazards in food products are discussed.
The book represents a concise state of the art addressing the current status
as well as various challenges for the development of new probiotics and
prebiotic-based foods and the niches for future research. All of the chapters
of the book have been contributed by a group of international contributors
who are well recognized and excellent in the frontier field of functional foods,
food microbiology, and microbial biotechnology. Together we have produced
an outstanding reference book that is expected to be a valuable resource for
researchers; teachers; students; food, nutrition, and health practitioners; and
all those working in the dairy, food, and nutraceutical industry. We extend
our sincere thanks to all the contributing authors whose co-operation has
made our task as editors a pleasure. We hope that this book will result in
enlightening and inspiring action for readers.
—Editors
Part I
Probiotics and Their Metabolites:
A Food Industry Perspective
CHAPTER 1
Action of Probiotic Microorganisms on

Mycotoxin Decontamination in Milk and
Milk Products
KATIA FRANCINE WOCHNER,* TÂNIA APARECIDA BECKER-ALGERI,
ELIANE COLLA, and DEISY ALESSANDRA DRUNKLER
*
Corresponding author. E-mail: katia_kfw29@hotmail.com
ABSTRACT
Milk is a foremost source of vital nutrients required for the development

plus human health maintenance. Nevertheless, it can also be a vehicle for
toxic agents, causing serious hazards in individuals who consume it, espe
cially children. Mycotoxins are among the main chemical contaminants
in milk, which are important secondary fungal metabolites in human and
animal health. When ingested by animals, they can be excreted in the
milk, and once present in milk, these mycotoxins can withstand most
treatments (pasteurization and ultra-high-temperature processing) to
obtain dairy products. Therefore, mycotoxins may be present in cheeses
and yogurts. It is imperative that milk and milk products should be free
from toxic components with regard to the high levels of consumption;
therefore, the study of efficient decontamination protocols that are safe
environmentally and precise is an important and current research topic.
Amongst the category of biological methods pertinent for the chemical
contaminants removal, many of the studies are mainly focused on
employing probiotic cultures, with promising outcomes. In this context,
the current chapter aims to review the literature on mycotoxins and their
decontamination processes from milk and milk products by means of
probiotics.
4 Microbial Biotechnology in Food Processing and Health
1.1 INTRODUCTION
There has been increasing concern related to the milk and dairy products
consumption because of contamination by mycotoxins, since, in recent
years, countless studies have demonstrated their incidence in fluid milk and
dairy products (Scaglioni et al., 2014; Bahrami et al., 2016; Becker-Algeri et
al., 2016; Michling et al., 2016; Al-Hilphy et al., 2016; Verma et al., 2017;
Kangethe et al., 2017; Zheng et al., 2017; Mao et al., 2018). Dairy products
consumption accounts for more than 80% of children’s dietary habits and
intake worldwide (Campagnollo et al., 2016). Mycotoxins are synthesized
as secondary metabolites by the molds of the genera Aspergillus, Fusarium,
and Penicillium, divided into six main classes: aflatoxins, ochratoxins
(OTAs), fumonisins, zearalenone (ZEA), trichothecenes, and ergot alkaloids
(Ahlberg et al., 2015). The structure of this secondary metabolites is depicted
in Figure 1.1.
FIGURE 1.1 Structure of six main classes of mycotoxins secondary metabolites synthesized
by the genera Aspergillus, Fusarium, and Penicillium.
Action of Probiotic Microorganisms on Mycotoxin Decontamination 5
The Food and Agriculture Organization (FAO) and the World Health
Organization (WHO) have emphasized mycotoxins in food because of the
great toxicological and carcinogenic potential that some of them pose to
animal and human health (International Agency for Research on Cancer
(IARC), 2012). Taking into account the high consumption of milk and dairy
products by the people and the risks to human health linked with the intake
of the most varied mycotoxins frequently present in them (Table 1.1), some
nations have established tolerance borders (Codex Alimentarius Commis
sion, 2001; Mercosul, 2002; European Commission, 2006). In addition,
socio-economic issues should be taken into account, since global legislation
is not compatible, as each country has its own regulations, which makes it
difficult to monitor, besides the lack of legislation for all milk-based products
manufactured and consumed around the world (Campagnollo et al., 2016).
TABLE 1.1 Mycotoxins of Milk and Dairy Products and it is Consumption Effects on
Human Health
Mycotoxins Chemical Nature Milk and Dairy Effects on Human
Products Health
Aflatoxin B1 Polyketide-coumarin Cheeses (Manchego, Acute toxicity,
(AFB1) (difurocoumaro Prato, Parmesan) immunosuppressive,
cyclopentenone) teratogenic,
mutagenic,
carcinogenic
Aflatoxin M1 Polyketide-coumarin Goat milk and Cow milk Immunotoxic,
(AFM1) (difurocoumaro (raw, pasteurized, ultra teratogenic,
cyclopentenone) high temperature UHT), mutagenic,
cream, butter, ice cream, hepatotoxic,
yogurt, cheeses (white carcinogenic
cheese, pickled cheese,
Tilsit, Brick, Parmesan,
Feta, Camembert,
Mozzarella, Manchego,
Kashar, Tulum, goat hard
cheese)
Andrastin A-D Meroterpenoids 21 brands of Blue-veined Impaired milk
cheeses (Danablue, production and
Castello, Klosterkrone, reproduction,
St. Clemens, Bleu inappetence,
D’auvergne, Castello, immunosuppression
Fourme d’Ambert,
Gorgonzola, Saint Agur
Bresse Bleu, Stilton, and
Magor)
TABLE 1.1 (Continued)

Products Health
Citrinin Polyketide Smear cheeses, surface Neurotoxicity,
(quinonemethine) mold semi-soft cheeses, hepatotoxicity
Prato, Parmesan, (occasionally),
goat soft cheese, and nephrotoxicity (acute
Saint Marcellin TM, and fatal)
(SoignonTM), Kashar
Cyclopiazonic Indole-tetramic acid White cheese, Immunotoxicity,
acid Camembert, Taleggio cytotoxicity, kodua
Italian, Kashar poisoning (acute
intoxication causing
nerve problems)
Isofumigacla Alkaloids Blue-veined cheeses Neurotoxic
vines A and B
Mycophenolic Phthalide German blue cheese, Immunosuppressive
acid Tilsit, Stilton, Roquefort, (A selective inhibitor
Bleu des Causses, Bleu of lymphocyte
d’Auvergne, Fourme proliferation)
d’Ambert, Manchego
Ochratoxin A Polyketide Cow milk-based infant Nephrotoxic,
(dihydroisocoumarin formulae, and Cheeses teratogenic, and
coupled with (different types of semi- immunotoxic,
L-phenylalanine) hard raw-milk cheese, decreasing
Gorgonzola, Roquefort). human growth
in malnourished
children. Endemic
toxicity-nephropathy
(Balkan region);
urothelial urinary
tract tumors
(Bulgaria) and
chronic interstitial
nephropathy (North
Africa)
Patulin Polyketide lactone Raw milk semi-hard Acute toxicity
cheese, Tulum, Blue- including
veined, Manchego neurotoxicity
(paralysis of motor
nerves, brain
damage, convulsion),
nephrotoxicity,
and hepatotoxicity,
teratogenicity
TABLE 1.1 (Continued)

Products Health
Penicillic acid Polyketide Cheddar, Tulum, Cytotoxic,
butyrolactone Roquefort, various hepatotoxic,
Swiss-type cheeses nephrotoxic,
genotoxic,
carcinogenesis
PR-toxin Eremophilane Various Blue-veined Carcinogenic,
sesquiterpene cheeses mutagenic, acute
toxicity (increase
of capillary
permeability and a
direct damage to the
heart, kidneys, lungs,
liver)
Roquefortine C Indole alkaloid Tulum, blue-white Neurotoxin (acute
(diketopiperazine) mold cheese (bovine) intoxication)
Manchego, Blue-veined
cheese (ewe, goat,
bovine, and mixture),
ripened blue, Roquefort
(ovine), Gorgonzola,
Stilton, Cabrales,
dressing of blue cheese
Source: Reprint from: Benkerroum (2016); with permission. © Elsevier.
There are different control measures for the exclusion of mycotoxins

from milk-based products; nonetheless, these processes do not provide
sufficient protection or are very costly to implement (El-Nezami et al., 1998;
Shcherbakova et al., 2015). In turn, the biological degradation of chemical
contaminants employing microbial cells or their metabolic products is a
specific alternative to reduce levels or eliminate these contaminants in food
while maintaining the food’s safety, since several foods contain microflora
added during their development (Ji et al., 2016; Sarlak et al., 2017). Several
lactic acid bacteria (LAB) and yeasts, because of their GRAS (generally
recognized as safe) standing as well as application as probiotics, are the most
studied to lessen the mycotoxins’ bioaccessibility in milk and its derivatives
(Bovo et al., 2013; Elsanhoty et al., 2014; Sarlak et al., 2017; Wochner et al.,
2018; Patel et al., 2018).
The aim of the current chapter is to examine the major mycotoxins

associated with milk and milk products and discusses the biological decon
tamination methods with the use of probiotic microorganisms.
1.2 MYCOTOXINS: AN OVERVIEW
Mycotoxins are among the most serious public health contaminants due
to their presence in food and the adverse effects they may cause in both
animals and humans (Benkerroum, 2016). They are a group of extremely
toxic secondary metabolites, which usually develop in places with extreme
conditions, such as low water availability, abrupt temperature variations,
and long periods of rain (Ahlberg et al., 2015). Contamination by myco
toxins occurs widely in foods of plant origin, all through their pre-and post
harvest or mainly during storage, especially in cereals, feed, fodder, and
other agricultural foodstuffs intended for animal or human consumption.
As said by the United Nations’ FAO, more than 25% of the food crops of
the world are appreciably polluted by means of mycotoxins (Campagnollo
et al., 2016). When ingested through contaminated food, mycotoxins are
usually transmitted to animal products including milk, egg, or meat, after
it get metabolized and biotransformed; most are chemically and thermally
stable in processing, including cooking, boiling, frying, and pasteurizing,
posing a potential risk, even in processed foods (Bruerton, 2001; Creppy et
al., 2002; Murphy et al., 2006).
Currently, more than 300 mycotoxins have been identified, and this number
continues to increase. Scientific attention is focused mainly on mycotoxins
that are carcinogenic and/or toxigenic, among which are aflatoxins, OTAs,
fumonisins, trichothecenes, and ZEA (Becker-Algeri et al., 2016). It is possible
to correlate many human diseases with an ingestion of mycotoxins, especially
the long-term consumption, the core lethal effects being carcinogenicity, neph
rotoxicity, hepatotoxicity, genotoxicity, dermal irritation, immunosuppression,
and reproductive disorders (Bovo et al., 2013; Lee and Ryu, 2015). According to
IARC (2012), aflatoxins such as aflatoxin B1 (AFB1) plus aflatoxin M1 (AFM1)
are basically classified as group 1 (carcinogenic to humans). Fumonisin B1
(FB1) and OTA belong to group 2B (most likely human carcinogen). Although
other toxins with ZEA and its derivatives (alpha-zeralanol, beta-zeralanol,
alpha-zearalenol, beta-zearalenol) are considered non-carcinogenic, they
cause other adverse effects, in particular, estrogenic effects, which may affect
reproduction in mammals (Huang et al., 2014).
Considering all mycotoxins present in foods, aflatoxin is the widely

distributed toxin in milk and food products and is expressed as the most toxic
among all known toxins (IARC, 2012). A total of 18 comparable compounds
are explained as aflatoxins, however the ones found naturally in foods are
six: B1 (AFB1), G1 (AFG1), M1 (AFM1), B2 (AFB2), G2 (AFG2), and M2
(AFM2) (Campagnollo et al., 2016; Patel et al., 2018). However, the foremost
prevalent type of form and the highest potent of among all mycotoxins are
AFB1 (Murphy et al., 2006; Corassin et al., 2013). These toxins are usually
soluble in organic solvents like benzene, chloroform, and methanol, have a
low molecular weight, are considered lipophilic molecules and are extremely
stable above 100°C temperatures (Murphy et al., 2006). It is important to
note that milk or dairy products can get contaminated through aflatoxin in
two ways: when lactating milk animals eat AFB1 polluted feed which will be
biotransformed in AFM1 and excreted in milk (indirect contamination), or
by direct contamination on milk-derived product as deliberate additives or
unintentional accidental contamination (Campagnollo et al., 2016).
The symptoms of aflatoxicosis (poisoning that results from ingesting afla
toxins) depend on the aflatoxin nature; the quantity ingested and the exposure
period; the sex, age, and health of the exposed person (Elsanhoty et al., 2014).
The main target organ is the liver due to the hydrophobic nature of aflatoxins,
once is a predominately lipophilic organ, and it is in hepatocytes where
each and every component that are transported through the blood stream are
accumulated and hence get concentrated (Campagnollo et al., 2016). Acute
aflatoxicosis where there is exposure to high-level of aflatoxins may result
in nausea and vomiting until serious liver damage such as cirrhosis or liver
failure, and death. Already the chronic aflatoxicosis can results in cancer,
especially liver cancer, immunosuppression, and interferes with metabolism
of protein as well as micronutrients that are vital to health (Bennet and Klich,
2003; Williams et al., 2004). Aflatoxins are of major concern in tropical and
subtropical climates, especially in developing countries where safe food
storage is not guaranteed (Aldars-García et al., 2016). Recently, the Centers
for Disease Control and Prevention reported cases of acute aflatoxicosis in
Kenya causing 125 deaths among 317 cases of poisoning caused by high levels
of AFB1 found in homegrown maize (CDC, 2004). Therefore, there is also a
great concern regarding the health of children major consumers of milk and
dairy products and whose sensitivity is remarkable and significantly higher as
compared to adults (Scaglioni et al., 2014). So, because of the regular utiliza
tion of milk or milk products at higher level, it is extremely essential that they
are free of poisonous ingredients (Flores-Flores et al., 2015).
1.2.1 MYCOTOXINS IN MILK AND MILK PRODUCTS
The incidence of mycotoxins in milk or dairy products has been accounted

for in different countries, demonstrating the global concern about these
toxins (Table 1.2).
TABLE 1.2 Incidence of Mycotoxins in Milk and Milk Products in Different Countries
Country Product Mycotoxin Number of Degree of References
Type Samples Contamination
(μg L–1/μg kg–1)
Argentina Raw milk AFM1 160 0.003–0.293 Michling et al.
(2016)
Brazil Cheese AFM1 58 0.01–0.3 Iha et al.
Yogurt 53 0.01–0.53 (2011)
Dairy drink 12 0.01–0.05
China Raw milk α-ZOL 30 0.024–0.073 Huang et al.
Pasteurized 12 0.036–0.045 (2014)
milk
Powdered 8 0.043–0.064
milk
Iran Traditional AFM1 360 0.05–0.30 Shahbazi et al.
cheese (2017)
Italy UHTa and raw FB1 10 0.26–0.43 Gazzoti et al.
milk (2009)
Mexico Pasteurized AFB1 290 0.05–0.42 Carvajal et al.
and UHTa milk (2003)
Serbia UHTa milk AFM1 438 0.025–1.00 Tomasevic et
Raw milk 678 0.025–>1.00 al. (2015)
White cheese 47 0.025–1.00
Yogurt 56 0.026–0.50
Note: UHT: Ultra-high temperature.
a
The mainly studied mycotoxin found in fluid milk and its derivatives
is aflatoxin AFM1, characterized as a hydroxylated derivative of AFB1 and
originating from the precursor’s ingestion by milch cattle by the consump
tion of feed, hay, and, mainly, contaminated silage. Hepatic biotransforma
tion occurs through enzymes of the cytochrome P450 complex partially
hydroxylated to AFM1 (Figure 1.2). According to Creppy (2002), in the milk,
about 0.3% to 6.2% of the entirety AFB1 ingested by the cattle are generally
converted into AFM1. The solubility of AFM1 in water facilitates its excre
tion in the biological fluids of animals, such as milk and urine, accumulating
in them (Oatley et al., 2000; Murphy et al., 2006).
Before some researchers investigated and confirmed AFB1 occurrence in
milk (Carvajal et al., 2003; Scaglioni et al., 2014), it was assumed that it
was fully biotransformed in other compounds, including its hydroxylated
AFM1. Due to the scarcity of these studies, only AFM1 is regulated and has
its maximum limits stipulated in milk-based food matrices. The European
Community and the Codex Alimentarius recommend a limit of 0.05 μg per
L in milk, milk powder, as well as other dairy products (Codex Alimentarius
Commission, 2001; European Commission, 2006). Other nations like the
United States of America (USA) and China set a boundary of 0.5 μg per L
for milk and dairy products. Brazil pursues the same limits established by the
Common Market of the South (Mercosul), who maximally permitted AFM1
concentrations in fluid milk, milk powder, and cheeses are 0.5 μg L–1, 5.0
μg per kg, and 2.5 μg per kg, respectively (Mercosul, 2002; Brasil, 2011).
On the other hand, there is still a lack of regulation of the definition of the
highest permitted levels of AFB1 in fluid milk and dairy products.
FIGURE 1.2 Hydroxylation of aflatoxin B1 in M1 by the P-450 system.
In addition to the problems related to the natural AFM1 contamination

of milk, further there is a great apprehension regarding the stability of
contaminant in foods (Iqbal et al., 2015), which includes thermal resistant
applied on the raw material during the manufacturing of dairy products,
such as pasteurization and sterilization, allowing the risk of this contamina
tion to be present also in products such as cheeses and yogurts (Tomasevic
et al., 2015; Li et al., 2017; Shahbazi et al., 2017). Furthermore, in some
cases, dairy products such as cheeses may present a higher concentration

(from three to five times greater) of AFM1 than in natural milk, because
AFM1 binds predominantly to the milk protein fraction and preferably in
hydrophobic regions of casein (Iha et al., 2011; Chavarría et al., 2017). In
milk-based products such as yogurt, the effect of fermentation reduces the
original concentrations of aflatoxin initially present in milk (Serrano-Niño et
al., 2013; Elsanhoty et al., 2014; Sarlak et al., 2017).
Few research articles show the incidence of other mycotoxins in milk
and derivatives, and unlike AFM1 there is no legislation for them. Gazzoti
et al. (2009); and Coffey et al. (2009) analyzed the mycotoxin FB1 pres
ence in milk, finding a mean level of 0.26 and 0.36 μg per L, respectively,
which, even at low concentrations, is worrisome, since the coexistence of
fumonisin and aflatoxin-mycotoxins could have toxicological effects and
synergistic carcinogens (Gelderblom et al., 2002). In Italy, Pattono et al.
(2011) found the incidence of mycotoxin OTA in samples of fluid milk
produced in organic farms, ranging between 0.07 and 0.110 µg L–1. Pattono
et al. (2013) analyzed the OTA mycotoxin in traditional cheeses produced
with raw milk, where six samples were contaminated with a concentra
tion of between 18.4 and 146.0 µg L–1 on the inside and between 1.0 and
262.2 µg L–1 on the outside. OTA is mainly linked with Balkan endemic
nephropathy (BEN) which is a fatal human renal syndrome (Bennet and
Klich, 2003). Moreover, when associated to AFM1, it presents a high
level of toxicity in human cells (Tavares et al., 2013). Huang et al. (2014)
detected mycotoxins ZEA, α-zearalenol (α-ZOL), and OTA, with their
respective mean concentrations 0.014, 0.024, and 0.056 μg per L in raw
milk; 0.025, 0.036, and 0.026 μg per L, respectively, in pasteurized milk;
and 0.011, 0.043, and 0.027 μg per L, respectively, in milk powder samples
collected from various milk farms and supermarkets in Beijing (China)
reinforcing the problem of the occurrence of other mycotoxins in milk and
dairy products.
1.3 BIOLOGICAL DECONTAMINATION OF MYCOTOXINS

IN MILK AND DAIRY PRODUCTS: USE OF PROBIOTIC
MICROORGANISMS
Since mycotoxins are present in milk like food products, their removal
can be a difficult step because of the several mycotoxins’ resistance to
tremendous ecological conditions (pH, temperature, radiation), in addition
to specific biological, chemical, and physical processes for their inhibition
(Benkerroum, 2016). There are different control measures for the removal
of mycotoxins in fluid milk plus related products; conversely, these methods
endow with an inadequate degree of protection or are difficult to implement
(El-Nezami et al., 1998; Shcherbakova et al., 2015). Mycotoxins’ biological
removal by making use of microorganisms (fungi, yeasts, and bacteria) and/
or enzymes is an alternative to reduce levels or eliminate mycotoxins in
foods, maintaining the safety and quality of mycotoxins (Sarlak et al., 2017;
Wochner et al., 2018).
Currently, attention has been paid to probiotics due to their ability to
reduce and/or inhibit the action of toxic compounds, including mycotoxins
(Bilandžić et al., 2011; Skrbic et al., 2014; Shaker and Elsharkawy, 2015;
Bahrami et al., 2016). Several species of LAB, bifidobacteria, and yeast
Saccharomyces cerevisiae var. boulardii, are considered as probiotics,
“living microorganisms which, when administered in suitable amounts, have
a beneficial effect on host health” (FAO/WHO, 2001). Although, the positive
actions linked with the regular probiotics consumption include improved
intestinal function by regulation of the endogenous beneficial microbes,
immunostimulation, increased bioaccessibility of nutrients (minerals and
vitamins), reduced symptoms of lactose intolerance, in addition to anticancer
and antimutagenic activity, acting in the colon cancer prevention and other
intestinal illnesses (Mallebrera et al., 2013: Shori, 2015).
Biological removal approach involves microbial cells (live or dead)
to eliminate mycotoxins, by cell wall absorbance, thus reducing their bio
accessibility (Kabak and Var, 2008; Serrano-Niño et al., 2013; Elsanhoty et
al., 2014). Bioaccessibility may be defined as ‘the amount of a compound
released from eaten food matrix into the gastrointestinal (GI) tract and made
accessible for consequent absorption by the intestine’ (Serrano-Niño et al.,
2013). The structure and strength of the microorganism-mycotoxin complex
are affected by various factors such as concentration, type, and specificity
of the microorganism, incubation temperature, pH, inactivation treatment,
nutrient addition, and food matrix (Bovo et al., 2013). The strains of micro
organisms most studied in in-vitro decontamination of mycotoxins involve
the genera of LAB such as Lactobacillus, Lactococcus, Streptococcus, and
Propionibacterium genera (Dalié et al., 2010; El-Khoury et al., 2011; Patel
et al., 2018). Besides bacteria, the yeast Saccharomyces cerevisiae has also
been successful in the detoxification of mycotoxins (Shetty and Jespersen,
2006; Armando et al., 2011; Karazhiyan et al., 2016).
Even though the exact mechanism of action for these microorganisms
on mycotoxins has not yet been known, the most relevant hypothesis is a
non-covalent physical adsorption of these microorganisms with the cell wall

components of bacterial cell or yeast cell, especially in specific peptidogly
cans and polysaccharides (Shetty and Jespersen, 2006). The peptidoglycan
layer contains mainly C-O, OH, and NH as functional groups and has been
reported as the main component involved in the biological adsorption of
mycotoxins (Zoghi et al., 2014). On the other hand, Serrano-Niño et al.
(2015) affirmed the significance of the teichoic acid during such interac
tion, where they suggested that the mode of interaction happened between
glucose and glycerol of the acid of the Lactobacillus strains examined with
the hydroxyl groups and with the carbonyl oxygen of AFB1. In addition, it is
believed that some treatments of inactivation, be they chemical, physical, or
enzymatic, are competent to enhance the aptitude of the microorganisms to
attach to the mycotoxin of the specific medium, since such disorders increase
the hydrophobic nature of the micro surface-organism, exposing the plasma
membrane components that were occupied when it was intact (Harkard et al.,
2001; Sarlak et al., 2017). The use of non-viable microbial cells is an advan
tage when fermentation is undesirable and the alteration of the product is not
desired. In addition, the different strains of LAB and S. cerevisiae yeasts are
not equivalent in terms of removal of toxin; in contrast, mycotoxin removal
aptitude is a feature of specific strains only, with their efficacy varying
distinctly. Such kinds of variations between strains propose dissimilarities
in cell wall or/and cell envelope structure as well as the existence of diverse
binding positions (Bovo, 2013; Karazhiyan et al., 2016).
El Khoury et al. (2011) evaluated the efficiency of AFM1 removal during
the production of yogurt (6 h) by cultures of Lactobacillus bulgaricus and
Streptococcus thermophilus, in isolation and together, obtaining a removal
of 58.5%, 37.7%, and 46.7% of mycotoxin, respectively. Using an in vitro
digestibility model, Serrano-Niño et al. (2013) evaluated the capability of
five probiotics (L. acidophilus, L. rhamnosus, L. johnsonii and Bifidobac
terium bifidum) to lower AFM1 aflatoxin bioaccessibility in milk that was
artificially spiked with it. The AFM1 bioaccessibility was decreased between
22.72 and 45.17%, depending on the used probiotic strain. Elsanhoty et
al. (2014) evaluated the prospective of five LAB strains (L. plantarum, L.
acidophilus, L. bulgaricus and S. thermophilus) and one strain of bifidobac
terium (Bif. angulatum) to eliminate AFM1 from fermented milk, yogurt.
The highest reduction found (87%) was the fermented test with 50% L.
plantarum and 50% yogurt (S. thermophilus and L. bulgaricus). Sarlak et
al. (2017) analyzed the effects of probiotic bacteria (L. casei, L. acidophilus,
L. rhamnosus, B. lactis) on the reduction of AFM1 in an established Iranian
fermented drinkable milk (Doogh), obtaining a result of 60.2% aflatoxin

reduction in 28 days of storage.
Taheur et al. (2017) investigated the biodegradation and/or adsorption
capacity of bacteria and yeasts isolated from kefir of Tunisia. The results
showed an adsorption of 82%, 94%, and 100% for the mycotoxins AFB1,
OTA, and ZEA, respectively; when kefir was grown in milk, with emphasis
on the L. kefiri strain KFLM3. These results propose that kefir consumption
may help to lessen mycotoxin toxic effects by reducing the GI absorption of
these. In addition to studies performed in a dairy matrix, there are studies
that demonstrate the potential for LAB decontamination in buffer solutions.
Sangsila et al. (2016) studied ZEA removal capability of 5 L. pentosus strains
from a sodium acetate buffer solution, also obtained good removal capaci
ties, ranging from 29.74 to 83.17%, indicating promise to also decrease the
contamination of mycotoxins from food.
The use of yeasts combined with LAB or even in isolation has also been
studied as a possible biological decontamination technique. Corassin et al.
(2013) assessed the aptitude of a strain of S. cerevisiae and a set of 3 bacteria
(Lactobacillus delbrueckii subsp. bulgaricus, L. rhamnosus, and B. lactis),
isolated or in mixture, to remove AFM1 from skimmed UHT milk samples.
The mean percentage of removal was 90% using isolated yeast, 11% for LAB,
and 100% at AFM1 levels when S. cerevisiae yeast was used in conjunction
with LAB. Karazhiyan et al. (2016) reported in their study a mean removal
of 75% of AFM1 in yogurt from the S. cerevisiae strain (PTCC 5177). For
comparison, viable and non-viable cells (acid, heat, and ultrasound) were
used. It was shown that the non-viable cells had a higher aptitude to bind
to aflatoxin and, consequently, to reduce it. For both Corassin et al. (2013);
and Karazhiyan et al. (2016), the aflatoxin removal method employing S.
cerevisiae alone or in conjunction with LAB, mainly strains used in several
food products, has prospective application to reduce AFM1 levels in milk and
other fermented foods such as yogurt.
1.4 SUMMARY AND CONCLUSION
Mycotoxin contamination in milk and dairy products plays a significant role

in food safety, due to ingestion in isolation or in combination, mainly in a
chronic way, related to serious health problems. The use of microorganisms,
such as LAB and some yeasts, is a promising approach for the safe, specific,
and effective decontamination of these foods. Several probiotic microor
ganisms have been studied and are of inquisitiveness for the expansion of
fermented dairy products intended for the consumption of human being and
currently have a novel probiotic attribute, the mycotoxin adsorption and
hence ultimately decontamination from food products.
KEYWORDS
• acid lactic bacteria

• aflatoxin
• bacteria cell wall
• Bifidobacterium spp.
• biological degradation
• contaminated milk
• dairy products
• decontamination methods
• fermentation
• food safety
• human health
• immunosuppression
• Lactobacillus spp.
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CHAPTER 2
New Technological Trends in Probiotics

Encapsulation for Their Stability
Improvement in Functional Foods and
Gastrointestinal Tract
MAJID NOOSHKAM* and ZAHRA ZAREIE
*
Corresponding author. E-mail: Nooshkamma@gmail.com
ABSTRACT
The ability of probiotics to alter gut community and render health-promoting

effect, induce food producers to use them as bioactive agents. However, they
are sensitive to adverse conditions that may encounter through production
and in the human stomach. As a matter of fact, researchers tried to increase
their bioavailability through various methods. Encapsulation is known as a
promising technique that has been frequently applied to protect and deliver
probiotic bacteria (i.e., Lactobacillus and Bifidobacterium species). The
present chapter discusses the encapsulation of probiotics in many delivery
systems such as emulsion, hydrogels, microgels, microcapsules, colloido
somes, and the Maillard reaction products (MRPs). It then highlights the
viability and bioavailability of the encapsulated probiotics in food products
and gastrointestinal (GI) tract.
2.1 INTRODUCTION
In recent years, in the light of significant demands for health-enhancing foods,

the concept of ‘functional foods’ has been emerged in the food industry to
fortify food products with bioactive ingredients having health benefits. In this
way, the incorporation of probiotic microorganisms into food is an outstanding
approach to produce functional foods. Probiotic strains have many docu
mented health-related properties and it is believed that the living LAB with
probiotic effect play an important and beneficial role in the intestinal tract of
the host (Heidebach et al., 2012). However, the loss of the number of living
probiotic cells (i.e., bioactivity loss) during production, storage, and gastroin
testinal (GI) transition is a key issue, and therefore several attempts have been
made to minimize the bioactivity loss of probiotics (Mattila-Sandholm et al.,
2002; Siuta-Cruce and Goulet, 2001; Shah, 2000).
In this area, the most prominent technique is encapsulation with potential
to significantly protect living probiotic cells or other microorganisms from
adverse conditions (Nualkaekul et al., 2013). Encapsulation has received
special attention in the food science to add functional ingredients, enzymes,
colorants, antioxidants, and microbial products into foods; in the medical
industry to produce drugs and vaccines; and in the tissue-engineered medical
products as scaffolds (Borges et al., 2012). Encapsulation is known as a
technology of packing compounds with solid, liquid, or gas nature in sealed
capsules to release them through a controlled mode in the right place or
time. The packed compounds or materials are commonly called fill, internal
phase, payload, actives, and core material; whilst, the coating material, wall
material, carrier, shell, capsule, and membrane are the terms used for the
packaging materials. It is worth to pointing out that polysaccharides (natural
or modified), proteins, lipids, synthetic polymers, gums, and sugars can be
employed as wall materials (Fang and Bhandari, 2010).
In general, the encapsulation technique is used in the pharmaceutical and
food sectors for different reasons; it is applied to: (i) decrease the reactivity of
core material to its environment and, in turn, lower its degradation rate; (ii)
lower the transfer/evaporation rate of core material to its surrounding environ
ment; (iii) provide an easier handling by modifying the physical properties
of the core material; (iv) separate the mixture components for lowering their
reactivity; (v) dilute the encapsulated compound when its low level is required;
(vi) cover or mask the unpleasant taste or flavor of the encapsulant; (vii)
ameliorate the food safety via suppressing the microbial proliferation; (viii)
boost water solubility of bioactive ingredients; and (ix) tailor the releasing
rate of the encapsulant for providing a slower release and in a controlled mode
(Cohen et al., 2011; Desai and Park, 2005; Wang et al., 2015; Zhu, 2017).
In probiotic encapsulation, the microbial cells are randomly embedded
and immobilized in a continuous matrix as core materials (Desai and Park,
2005), and therefore the terms “encapsulation,” “immobilization,” and
“entrapment” are commonly used as synonyms in this regard (Krasaekoopt
et al., 2003; Anal and Singh, 2007). Different microorganisms have been
encapsulated in biocompatible and semipermeable materials capable of
New Technological Trends in Probiotics Encapsulation 23
modulating the delivery of probiotic cells (Gbassi and Vandamme, 2012).

The aim of probiotic encapsulation is to decrease the loss of living cell
numbers in foods and finally in the human gut. In addition, the encapsulated
probiotic should be released completely from the capsule in the intestinal
tract. It is noteworthy that the size of the capsule should be remarkably small
to prevent adverse sensorial properties on the functional foods containing
probiotics (Naidu et al., 1999).
This chapter reviews the importance of probiotic microorganisms in food
systems, factor influencing their bioactivity, and different approaches of
probiotic encapsulation. In addition, the stability of the encapsulated probi
otics in food products and gastrointestinal tract (GIT) has been discussed.
2.2 THE IMPORTANCE OF PROBIOTICS IN FOOD SYSTEMS
In the last decades, there has been a great deal of attention in the beneficial role
of probiotic microorganisms in human health. Probiotics render their positive
health effects when they reach to the action sites alive and proliferate in certain
numbers. It has been recommended by the International Dairy Federation that
the probiotic bacteria should be present at least 107 CFU g–1 in active form in
the product to exert their health-promoting effects (Sultana et al., 2000).
Probiotics are recognized as live microbial supplements with the ability
to improve the microbial balance of the host intestine. They have many
claimed benefits such as, controlling cholesterol levels, inhibition of the
growth of food pathogenic and poisoning bacteria in the digestive tract, anti-
carcinogenic effect, acid production, bacteriocin formation, the improve
ment of immune system, and the production of β-galactosidase with lactose
hydrolyzing activity, suitable for people suffering from lactose intolerance
(Krasaekoopt et al., 2003).
The positive effects are dependent on the survivability and proliferation
of the probiotic strains in the GI of the host. Furthermore, most products
containing probiotics on the market have a very short shelf-life, do not
usually contain the required viable cell numbers to possess a prebiotic
effect, and viable cells content decreases significantly after consumption
due to bile secretions and acidic environment in the GI (Todorov et al.,
2012). The survivability of probiotic cells in food products is influenced
by many factors include storage temperature, pH, H2O2 production, and
post-acidification during the fermentation process. Hence, the protection of
probiotic bacteria within a physical barrier against harsh circumstances is
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And give away ere nightfall all he made.
“'Can I have lodging here?' once more I said.

He blew a whiff, and, leaning back his head,
'You come a piece through Bailey’s woods, I s’pose,
Acrost a bridge where a big swamp-oak grows?
It don’t grow, neither; it’s ben dead ten year,
Nor th' ain’t a livin' creetur, fur nor near,
Can tell wut killed it; but I some misdoubt
’Twas borers, there’s sech heaps on ’em about.
You did n' chance to run ag’inst my son,
A long, slab-sided youngster with a gun?
He’d oughto ben back more ’n an hour ago,
An' brought some birds to dress for supper—sho!
There he comes now. 'Say, Obed, wut ye got?
(He’ll hev some upland plover like as not.)
Wal, them’s real nice uns, an’ll eat A 1,
Ef I can stop their bein' over-done;
Nothin' riles me (I pledge my fastin' word)
Like cookin' out the natur' of a bird;
(Obed, you pick ’em out o' sight an' sound,
Your ma’am don’t love no feathers cluttrin' round;)
Jes' scare ’em with the coals,—thet’s my idee.'
Then, turning suddenly about on me,
'Wal, Square, I guess so. Callilate to stay?
I’ll ask Mis' Weeks; ’bout thet it’s hern to say.'
“Well, there I lingered all October through,

In that sweet atmosphere of hazy blue,
So leisurely, so soothing, so forgiving,
That sometimes makes New England fit for living.
I watched the landscape, erst so granite glum,
Bloom like the south side of a ripening plum,
And each rock-maple on the hillside make
His ten days' sunset doubled in the lake;
The very stone walls draggling up the hills
Seemed touched, and wavered in their roundhead wills.
Ah! there’s a deal of sugar in the sun!
Tap me in Indian summer, I should run
A juice to make rock-candy of,—but then
We get such weather scarce one year in ten.
“There was a parlor in the house, a room

To make you shudder with its prudish gloom.
The furniture stood round with such an air,
There seemed an old maid’s ghost in every chair,
Which looked as it had scuttled to its place
And pulled extempore a Sunday face,
Too smugly proper for a world of sin,
Like boys on whom the minister comes in.
The table, fronting you with icy stare,
Strove to look witless that its legs were bare,
While the black sofa with its horse-hair pall
Gloomed like a bier for Comfort’s funeral.
Each piece appeared to do its chilly best
To seem an utter stranger to the rest,
As if acquaintanceship were deadly sin,
Like Britons meeting in a foreign inn.
Two portraits graced the wall in grimmest truth,
Mister and Mistress W. in their youth,—
New England youth, that seems a sort of pill,
Half wish-I-dared, half Edwards on the Will,
Bitter to swallow, and which leaves a trace
Of Calvinistic cholic on the face.
Between them, o’er the mantel, hung in state
Solomon’s temple, done in copperplate;
Invention pure, but meant, we may presume,
To give some Scripture sanction to the room
To give some Scripture sanction to the room.
Facing this last, two samplers you might see,
Each, with its urn and stiffly-weeping tree,
Devoted to some memory long ago
More faded than their lines of worsted woe;
Cut paper decked their frames against the flies,
Though none e’er dared an entrance who were wise,
And bushed asparagus in fading green
Added its shiver to the franklin clean.
“When first arrived, I chilled a half-hour there,

Nor dared deflower with use a single chair;
I caught no cold, yet flying pains could find
For weeks in me,—a rheumatism of mind.
One thing alone imprisoned there had power
To hold me in the place that long half-hour:
A scutcheon this, a helm-surmounted shield,
Three griffins argent on a sable field;
A relic of the shipwrecked past was here,
And Ezra held some Old-World lumber dear.
Nay, do not smile; I love this kind of thing,
These cooped traditions with a broken wing,
This freehold nook in Fancy’s pipe-blown ball,
This less than nothing that is more than all!
Have I not seen sweet natures kept alive
Amid the humdrum of your business hive,
Undowered spinsters shielded from all harms,
By airy incomes from a coat of arms?”
He paused a moment, and his features took

The flitting sweetness of that inward look
I hinted at before; but, scarcely seen,
It shrank for shelter ’neath his harder mien,
And, rapping his black pipe of ashes clear,
He went on with a self-derisive sneer:
“No doubt we make a part of God’s design,
And break the forest-path for feet divine;
To furnish foothold for this grand prevision
Is good and yet to be the mere transition
Is good, and yet—to be the mere transition,
That, you will say, is also good, though I
Scarce like to feed the ogre By-and-by.
Raw edges rasp my nerves; my taste is wooed
By things that are, not going to be, good,
Though were I what I dreamed two lustres gone,
I’d stay to help the Consummation on,
Whether a new Rome than the old more fair,
Or a deadflat of rascal-ruled despair;
But my skull somehow never closed the suture
That seems to knit yours firmly with the future,
So you ’ll excuse me if I’m sometimes fain
To tie the past’s warm nightcap o’er my brain;
I’m quite aware ’tis not in fashion here,
But then your northeast winds are so severe!
“But to my story: though ’tis truly naught

But a few hints in Memory’s sketchbook caught,
And which may claim a value on the score
Of calling back some scenery now no more.
Shall I confess? The tavern’s only Lar
Seemed (be not shocked!) its homely-featured bar.
Here dozed a fire of beechen logs, that bred
Strange fancies in its embers golden-red,
And nursed the loggerhead whose hissing dip,
Timed by nice instinct, creamed the mug of flip
That made from mouth to mouth its genial round,
Nor left one nature wholly winter-bound;
Hence dropt the tinkling coal all mellow-ripe
For Uncle Reuben’s talk-extinguished pipe;
Hence rayed the heat, as from an in-door sun,
That wooed forth many a shoot of rustic fun.
Here Ezra ruled as king by right divine;
No other face had such a wholesome shine,
No laugh like his so full of honest cheer;
Above the rest it crowed like Chanticleer.
“In this one room his dame you never saw,

Wh i db t ld S li l
Where reigned by custom old a Salic law;
Here coatless lolled he on his throne of oak,
And every tongue paused midway if he spoke.
Due mirth he loved, yet was his sway severe;
No blear-eyed driveller got his stagger here;
'Measure was happiness; who wanted more,
Must buy his ruin at the Deacon’s store;'
None but his lodgers after ten could stay,
Nor after nine on eves of Sabbath-day.
He had his favorites and his pensioners,
The same that gypsy Nature owns for hers:
Loose-ended souls, whose skills bring scanty gold,
And whom the poor-house catches when they ’re old;
Rude country-minstrels, men who doctor kine,
Or graft, and, out of scions ten, save nine;
Creatures of genius they, but never meant
To keep step with the civic regiment.
These Ezra welcomed, feeling in his mind
Perhaps some motions of the vagrant kind;
These paid no money, yet for them he drew
Special Jamaica from a tap they knew,
And, for their feelings, chalked behind the door
With solemn face a visionary score.
This thawed to life in Uncle Reuben’s throat
A torpid shoal of jest and anecdote,
Like those queer fish that doze the droughts away,
And wait for moisture, wrapt in sun-baked clay;
This warmed the one-eyed fiddler to his task,
Perched in the corner on an empty cask,
By whose shrill art rapt suddenly, some boor
Rattled a double-shuffle on the floor;
'Hull’s Victory' was, indeed, the favorite air,
Though 'Yankee Doodle' claimed its proper share.
“'Twas there I caught from Uncle Reuben’s lips,

In dribbling monologue ’twixt whiffs and sips,
The story I so long have tried to tell;
The humor coarse, the persons common,—well,
From Nature only do I love to paint,
Whether she send a satyr or a saint;
To me Sincerity’s the one thing good,
Soiled though she be and lost to maidenhood.
Quompegan is a town some ten miles south
From Jethro, at Nagumscot river-mouth,
A seaport town, and makes its title good
With lumber and dried fish and eastern wood.
Here Deacon Bitters dwelt and kept the Store,
The richest man for many a mile of shore;
In little less than everything dealt he,
From meeting-houses to a chest of tea;
So dextrous therewithal a flint to skin,
He could make profit on a single pin;
In business strict, to bring the balance true
He had been known to bite a fig in two,
And change a board-nail for a shingle-nail.
All that he had he ready held for sale,
His house, his tomb, whate’er the law allows,
And he had gladly parted with his spouse.
His one ambition still to get and get,
He would arrest your very ghost for debt.
His store looked righteous, should the Parson come,
But in a dark back-room he peddled rum,
And eased Ma’am Conscience, if she e’er would scold,
By christening it with water ere he sold.
A small, dry man he was, who wore a queue,
And one white neckcloth all the week-days through,—
On Monday white, by Saturday as dun
As that worn homeward by the prodigal son.
His frosted earlocks, striped with foxy brown,
Were braided up to hide a desert crown;
His coat was brownish, black perhaps of yore;
In summer-time a banyan loose he wore;
His trousers short, through many a season true,
Made no pretence to hide his stockings blue;
A waistcoat buff his chief adornment was,
Its porcelain buttons rimmed with dusky brass
Its porcelain buttons rimmed with dusky brass.
A deacon he, you saw it in each limb,
And well he knew to deacon-off a hymn,
Or lead the choir through all its wandering woes
With voice that gathered unction in his nose,
Wherein a constant snuffle you might hear,
As if with him ’twere winter all the year.
At pew-head sat he with decorous pains,
In sermon-time could foot his weekly gains,
Or, with closed eyes and heaven-abstracted air,
Could plan a new investment in long-prayer.
A pious man, and thrifty too, he made
The psalms and prophets partners in his trade,
And in his orthodoxy straitened more
As it enlarged the business at his store;
He honored Moses, but, when gain he planned,
Had his own notion of the Promised Land.
“Soon as the winter made the sledding good,

From far around the farmers hauled him wood,
For all the trade had gathered ’neath his thumb.
He paid in groceries and New England rum,
Making two profits with a conscience clear,—
Cheap all he bought, and all he paid with dear.
With his own mete-wand measuring every load,
Each somehow had diminished on the road;
An honest cord in Jethro still would fail
By a good foot upon the Deacon’s scale,
And, more to abate the price, his gimlet eye
Would pierce to cat-sticks that none else could spy;
Yet none dared grumble, for no farmer yet
But New Year found him in the Deacon’s debt.
“While the first snow was mealy under feet,

A team drawled creaking down Quompegan street.
Two cords of oak weighed down the grinding sled,
And cornstalk fodder rustled overhead;
The oxen’s muzzles, as they shouldered through,
Were silver fringed; the driver’s own was blue
Were silver-fringed; the driver s own was blue
As the coarse frock that swung below his knee.
Behind his load for shelter waded he;
His mittened hands now on his chest he beat,
Now stamped the stiffened cowhides of his feet,
Hushed as a ghost’s; his armpit scarce could hold
The walnut whipstock slippery-bright with cold.
What wonder if, the tavern as he past,
He looked and longed, and stayed his beasts at last,
Who patient stood and veiled themselves in steam
While he explored the bar-room’s ruddy gleam?
“Before the fire, in want of thought profound,

There sat a brother-townsman weather-bound:
A sturdy churl, crisp-headed, bristly-eared,
Red as a pepper; ’twixt coarse brows and beard
His eyes lay ambushed, on the watch for fools,
Clear, gray, and glittering like two bay-edged pools;
A shifty creature, with a turn for fun,
Could swap a poor horse for a better one,—
He’d a high-stepper always in his stall;
Liked far and near, and dreaded therewithal.
To him the in-comer, 'Perez, how d’ye do?'
'Jest as I’m mind to, Obed; how do you?'
Then, his eyes twinkling such swift gleams as run
Along the levelled barrel of a gun
Brought to his shoulder by a man you know
Will bring his game down, he continued, 'So,
I s’pose you’re haulin' wood? But you’re too late;
The Deacon’s off; Old Splitfoot couldn’t wait;
He made a bee-line las' night in the storm
To where he won’t need wood to keep him warm.
’Fore this he’s treasurer of a fund to train
Young imps as missionaries; hopes to gain
That way a contract that he has in view
For fireproof pitchforks of a pattern new.
It must have tickled him, all drawbacks weighed,
To think he stuck the Old One in a trade;
His soul, to start with, wasn’t worth a carrot,
And all he’d left ’ould hardly serve to swear at.'
“By this time Obed had his wits thawed out,

And, looking at the other half in doubt,
Took off his fox-skin cap to scratch his head,
Donned it again, and drawled forth, 'Mean he’s dead?'
'Jesso; he’s dead and t’other d that follers
With folks that never love a thing but dollars.
He pulled up stakes last evening, fair and square,
And ever since there’s been a row Down There.
The minute the old chap arrived, you see,
Comes the Boss-devil to him, and says he,
“What are you good at? Little enough, I fear;
We callilate to make folks useful here.”
“Well,” says old Bitters, “I expect I can
Scale a fair load of wood with e’er a man.”
“Wood we don’t deal in; but perhaps you’ll suit,
Because we buy our brimstone by the foot:
Here, take this measurin'-rod, as smooth as sin,
And keep a reckonin' of what loads comes in.
You’ll not want business, for we need a lot
To keep the Yankees that you send us hot;
At firin' up they’re barely half as spry
As Spaniards or Italians, though they’re dry;
At first we have to let the draught on stronger,
But, heat ’em through, they seem to hold it longer.”
“'Bitters he took the rod, and pretty soon

A teamster comes, whistling an ex-psalm tune.
A likelier chap you wouldn’t ask to see,
No different, but his limp, from you or me'—
'No different, Perez! Don’t your memory fail?
Why, where in thunder was his horns and tail?'
'They’re only worn by some old-fashioned pokes;
They mostly aim at looking just like folks.
Sech things are scarce as queues and top-boots here;
’Twould spoil their usefulness to look too queer.
p q
Ef you could always know ’em when they come,
They’d get no purchase on you: now be mum.
On come the teamster, smart as Davy Crockett,
Jinglin' the red-hot coppers in his pocket,
And clost behind, ('twas gold-dust, you’d ha' sworn,)
A load of sulphur yallower’n seed-corn;
To see it wasted as it is Down There
Would make a Friction-Match Co. tear its hair!
“Hold on!” says Bitters, “stop right where you be;
You can’t go in athout a pass from me.”
“All right,” says t’other, “only step round smart;
I must be home by noon-time with the cart.”
Bitters goes round it sharp-eyed as a rat,
Then with a scrap of paper on his hat
Pretends to cipher. “By the public staff,
That load scarce rises twelve foot and a half.”
“There’s fourteen foot and over,” says the driver,
“Worth twenty dollars, ef it’s worth a stiver;
Good fourth-proof brimstone, that’ll make ’em squirm,—
I leave it to the Headman of the Firm;
After we masure it, we always lay
Some on to allow for settlin' by the way.
Imp and full-grown, I’ve carted sulphur here,
And given fair satisfaction, thirty year.”
With that they fell to quarrellin' so loud
That in five minutes they had drawed a crowd,
And afore long the Boss, who heard the row,
Comes elbowin' in with “What’s to pay here now?”
Both parties heard, the measurin'-rod he takes,
And of the load a careful survey makes.
“Sence I’ve bossed the business here,” says he,
“No fairer load was ever seen by me.”
Then, turnin' to the Deacon, “You mean cus,
None of your old Quompegan tricks with us!
They won’t do here: we’re plain old-fashioned folks,
And don’t quite understand that kind o' jokes.
I know this teamster, and his pa afore him,
And the hard working Mrs D that bore him;
And the hard-working Mrs. D. that bore him;
He wouldn’t soil his conscience with a lie,
Though he might get the custom-house thereby.
Here, constable, take Bitters by the queue,
And clap him into furnace ninety-two,
And try this brimstone on him; if he’s bright,
He’ll find the masure honest afore night.
He isn’t worth his fuel, and I’ll bet
The parish oven has to take him yet!” ’
“This is my tale, heard twenty years ago

From Uncle Reuben, as the logs burned low,
Touching the walls and ceiling with that bloom
That makes a rose’s calyx of a room.
I could not give his language, wherethrough ran
The gamy flavor of the bookless man
Who shapes a word before the fancy cools,
As lonely Crusoe improvised his tools.
I liked the tale,—’twas like so many told
By Rutebeuf and his brother Trouvères bold;
Nor were the hearers much unlike to theirs,
Men unsophisticate, rude-nerved as bears.
Ezra is gone and his large-hearted kind,
The landlords of the hospitable mind;
Good Warriner of Springfield was the last;
An inn is now a vision of the past;
One yet-surviving host my mind recalls,—
You’ll find him if you go to Trenton Falls.”
THE ORIGIN OF DIDACTIC POETRY.

When wise Minerva still was young
And just the least romantic,
Soon after from Jove’s head she flung
That preternatural antic,
’Tis said, to keep from idleness
Or flirting, those twin curses,
She spent her leisure, more or less,
In writing po——, no, verses.
How nice they were! to rhyme with far

A kind star did not tarry;
The metre, too, was regular
As schoolboy’s dot and carry;
And full they were of pious plums,
So extra-super-moral,—
For sucking Virtue’s tender gums
Most tooth-enticing coral.
A clean, fair copy she prepares,

Makes sure of moods and tenses,
With her own hand,—for prudence spares
A man-(or woman-)-uensis;
Complete, and tied with ribbons proud,
She hinted soon how cosy a
Treat it would be to read them loud
After next day’s Ambrosia.
The Gods thought not it would amuse

So much as Homer’s Odyssees,
But could not very well refuse
The properest of Goddesses;
So all sat round in attitudes
Of various dejection,
As with a hem! the queen of prudes
Began her grave prelection.
At the first pause Zeus said, “Well sung!—

I k Ph b h k ”
I mean—ask Phœbus,—he knows.”
Says Phœbus, “Zounds! a wolf’s among
Admetus’s merinos!
Fine! very fine! but I must go;
They stand in need of me there;
Excuse me!” snatched his stick, and so
Plunged down the gladdened ether.
With the next gap, Mars said, “For me

Don’t wait,—naught could be finer,
But I’m engaged at half past three,—
A fight in Asia Minor!”
Then Venus lisped, “I’m sorely tried,
These duty-calls are vip’rous;
But I must go; I have a bride
To see about in Cyprus.”
Then Bacchus,—“I must say good bye,

Although my peace it jeopards;
I meet a man at four, to try
A well-broke pair of leopards.”
His words woke Hermes. “Ah!” he said,
“I so love moral theses!”
Then winked at Hebe, who turned red,
And smoothed her apron’s creases.
Just then Zeus snored,—the Eagle drew

His head the wing from under;
Zeus snored,—o’er startled Greece there flew
The many-volumed thunder.
Some augurs counted nine, some, ten;
Some said ’twas war, some, famine,
And all, that other-minded men
Would get a precious——.
Proud Pallas sighed, “It will not do;

Against the Muse I’ve sinned, oh!”
And her torn rhymes sent flying through
Ol ’ b k i d
Olympus’s back window.
Then, packing up a peplus clean,
She took the shortest path thence,
And opened, with a mind serene,
A Sunday-school in Athens.
The verses? Some in ocean swilled,

Killed every fish that bit to ’em;
Some Galen caught, and, when distilled,
Found morphine the residuum;
But some that rotted on the earth
Sprang up again in copies,
And gave two strong narcotics birth,
Didactic verse and poppies.
Years after, when a poet asked

The Goddess’s opinion,
As one whose soul its wings had tasked
In Art’s clear-aired dominion,
“Discriminate,” she said, “betimes;
The Muse is unforgiving;
Put all your beauty in your rhymes,
Your morals in your living.”
THE FLYING DUTCHMAN.
Don’t believe in the Flying Dutchman?

I’ve known the fellow for years;
My button I’ve wrenched from his clutch, man:
I shudder whenever he nears!
He’s a Rip van Winkle skipper,

A Wandering Jew of the sea,
Who sails his bedevilled old clipper
In the wind’s eye, straight as a bee.
Back topsails! you can’t escape him;

The man-ropes stretch with his weight,
And the queerest old toggeries drape him,
The Lord knows how long out of date!
Like a long-disembodied idea,

(A kind of ghost plentiful now,)
He stands there; you fancy you see a
Coeval of Teniers or Douw.
He greets you; would have you take letters:

You scan the addresses with dread,
While he mutters his donners and wetters,—
They’re all from the dead to the dead!
You seem taking time for reflection,

But the heart fills your throat with a jam,
As you spell in each faded direction
An ominous ending in dam.
Am I tagging my rhymes to a legend?

That were changing green turtle to mock:
No, thank you! I’ve found out which wedge-end
Is meant for the head of a block.
The fellow I have in my mind’s eye

Plays the old Skipper’s part here on shore,
And sticks like a burr, till he finds I
Have got just the gauge of his bore.
This postman ’twixt one ghost and t’other,

With last dates that smell of the mould,
I have met him (O man and brother,
Forgive me!) in azure and gold.
In the pulpit I’ve known of his preaching,

Out of hearing behind the time,
Some statement of Balaam’s impeaching,
Giving Eve a due sense of her crime.
I have seen him some poor ancient thrashing
Into something (God save us!) more dry,
With the Water of Life itself washing
The life out of earth, sea, and sky.
O dread fellow-mortal, get newer

Despatches to carry, or none!
We’re as quick as the Greek and the Jew were
At knowing a loaf from a stone.
Till the couriers of God fail in duty,

We sha’n’t ask a mummy for news,
Nor sate the soul’s hunger for beauty
With your drawings from casts of a Muse.
CREDIDIMUS JOVEM REGNARE.
O days endeared to every Muse,

When nobody had any Views,
Nor, while the cloudscape of his mind
By every breeze was new designed,
Insisted all the world should see
Camels or whales where none there be!
O happy days, when men received
From sire to son what all believed,
And left the other world in bliss,
Too busy with bedevilling this!
Beset by doubts of every breed

In the last bastion of my creed,
With shot and shell for Sabbath-chime,
I watch the storming-party climb,
Panting (their prey in easy reach),
To pour triumphant through the breach
In walls that shed like snowflakes tons
Of missiles from old-fashioned guns,
B t bl ’ th th t th t
But crumble ’neath the storm that pours
All day and night from bigger bores.
There, as I hopeless watch and wait
The last life-crushing coil of Fate,
Despair finds solace in the praise
Of those serene dawn-rosy days
Ere microscopes had made us heirs
To large estates of doubts and snares,
By proving that the title-deeds,
Once all-sufficient for men’s needs,
Are palimpsests that scarce disguise
The tracings of still earlier lies,
Themselves as surely written o’er
An older fib erased before.
So from these days I fly to those

That in the landlocked Past repose,
Where no rude wind of doctrine shakes
From bloom-flushed boughs untimely flakes;
Where morning’s eyes see nothing strange,
No crude perplexity of change,
And morrows trip along their ways
Secure as happy yesterdays.
Then there were rulers who could trace
Through heroes up to gods their race,
Pledged to fair fame and noble use
By veins from Odin filled or Zeus,
And under bonds to keep divine
The praise of a celestial line.
Then priests could pile the altar’s sods,
With whom gods spake as they with gods,
And everywhere from haunted earth
Broke springs of wonder, that had birth
In depths divine beyond the ken
And fatal scrutiny of men;
Then hills and groves and streams and seas
Thrilled with immortal presences,
Not too ethereal for the scope
Of human passion’s dream or hope.
Now Pan at last is surely dead,

And King No-Credit reigns instead,
Whose officers, morosely strict,
Poor Fancy’s tenantry evict,
Chase the last Genius from the door,
And nothing dances any more.
Nothing? Ah, yes, our tables do,
Drumming the Old One’s own tattoo,
And, if the oracles are dumb,
Have we not mediums? Why be glum?
Fly thither? Why, the very air

Is full of hindrance and despair!
Fly thither? But I cannot fly;
My doubts enmesh me if I try,—
Each lilliputian, but, combined,
Potent a giant’s limbs to bind.
This world and that are growing dark;
A huge interrogation mark,
The Devil’s crook episcopal,
Still borne before him since the Fall,
Blackens with its ill-omened sign
The old blue heaven of faith benign.
Whence? Whither? Wherefore? How? Which? Why?
All ask at once, all wait reply.
Men feel old systems cracking under ’em;
Life saddens to a mere conundrum
Which once Religion solved, but she
Has lost—has Science found?—the key.
What was snow-bearded Odin, trow,

The mighty hunter long ago,
Whose horn and hounds the peasant hears
Still when the Northlights shake their spears?
Science hath answers twain, I’ve heard;
Choose which you will, nor hope a third;
y p
Whichever box the truth be stowed in,
There’s not a sliver left of Odin.
Either he was a pinchbrowed thing,
With scarcely wit a stone to fling,
A creature both in size and shape
Nearer than we are to the ape,
Who hung sublime with brat and spouse
By tail prehensile from the boughs,
And, happier than his maimed descendants,
The culture-curtailed independents,
Could pluck his cherries with both paws,
And stuff with both his big-boned jaws;
Or else the core his name enveloped
Was from a solar myth developed,
Which, hunted to its primal shoot,
Takes refuge in a Sanskrit root,
Thereby to instant death explaining
The little poetry remaining.
Try it with Zeus, ’tis just the same;

The thing evades, we hug a name;
Nay, scarcely that,—perhaps a vapor
Born of some atmospheric caper.
All Lempriere’s fables blur together
In cloudy symbols of the weather,
And Aphrodite rose from frothy seas
But to illustrate such hypotheses.
With years enough behind his back,
Lincoln will take the selfsame track,
And prove, hulled fairly to the cob,
A mere vagary of Old Prob.
Give the right man a solar myth,
And he’ll confute the sun therewith.
They make things admirably plain,

But one hard question will remain:
If one hypothesis you lose,
Another in its place you choose,

Microbial Biotechnology in Food Processing and Health: Advances, Challenges, and Potential 1st Edition Deepak Kumar Verma (Editor)

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Microbial Biotechnology in Food Processing and Health: Advances, Challenges, and Potential 1st Edition Deepak Kumar Verma (Editor)

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Microbial Biotechnology in Food

Processing and Health: Advances,

Food Processing Advances in Thermal Technologies

Food Processing Advances In Non Thermal Technologies

Advances in Probiotics: Microorganisms in Food and

Advances in Fisheries Biotechnology Pramod Kumar Pandey

Environmental Pollution and Remediation Environmental

Microbial Technologies for Wastewater Recycling and

Cambridge IGCSE and O Level History Workbook 2C - Depth

Agro-Processing and Food Engineering: Operational and

© 2023 by Apple Academic Press, Inc.

Library and Archives Canada Cataloguing in Publication

CIP data on file with US Library of Congress

ISBN: 978-1-77463-728-9 (hbk)

Deepak Kumar Verma, PhD

Deepak Kumar Verma, PhD, is an agricultural

Ami R. Patel, PhD

Ami R. Patel, PhD, is an Assistant Professor in

Sudhanshu Billoria, PhD

Sudhanshu Billoria, PhD, is an Assistant

Geetanjali Kaushik, PhD

Geetanjali Kaushik, PhD, is an Associate

Maninder Kaur, PhD

Maninder Kaur, PhD, is working as an Associate

Part I: Probiotics and Their Metabolites: A Food Industry Perspective .......... 1

2. New Technological Trends in Probiotics Encapsulation for

3. Bioactive Peptides (BPs) as Functional Foods: Production Process,

Part II: Probiotics and Potential Health Benefits ............................................. 79

5. Ice Cream as Probiotic Food and Its Potential Benefits in

Part III: Industrial Aspects of Microbial Food Technologies ........................ 165

7. Innovation in Technology for Processing of Sheep and

8. Sustainable Use of Food-Grade Microorganisms in

9. Advanced Molecular Tools and Techniques for Assessment of

10. Industrial Prospects of Bacterial Microcompartment Technologies..... 297

11. Management of Microbiological Hazards in the

Index .................................................................................................................... 337

Carlos Eduardo Barão

Adriano Gomes da Cruz

Deisy Alessandra Drunkler

Suellen Jensen Klososki

Vanessa Aparecida Marcolino

Vanessa Guimarães Alves Olher

Antonio José Trujillo

Deepak Kumar Verma

Katia Francine Wochner

ACE angiotensin converting enzyme

GFP green fluorescent protein

YPD yeast extract potato dextrose agar

technologies dealing with assessing microbial diversities or management of

Action of Probiotic Microorganisms on

Milk is a foremost source of vital nutrients required for the development

TABLE 1.1 (Continued)

TABLE 1.1 (Continued)

There are different control measures for the exclusion of mycotoxins

The aim of the current chapter is to examine the major mycotoxins

1.2 MYCOTOXINS: AN OVERVIEW

Considering all mycotoxins present in foods, aflatoxin is the widely

1.2.1 MYCOTOXINS IN MILK AND MILK PRODUCTS

The incidence of mycotoxins in milk or dairy products has been accounted

FIGURE 1.2 Hydroxylation of aflatoxin B1 in M1 by the P-450 system.

In addition to the problems related to the natural AFM1 contamination

cases, dairy products such as cheeses may present a higher concentration

1.3 BIOLOGICAL DECONTAMINATION OF MYCOTOXINS