Microbiology in Dairy Processing E-Book
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- Herausgeber: John Wiley & Sons
- Kategorie: Fachliteratur
- Serie: Institute of Food Technologists Series
- Sprache: Englisch
An authoritative guide to microbiological solutions to common challenges encountered in the industrial processing of milk and the production of milk products
Microbiology in Dairy Processing offers a comprehensive introduction to the most current knowledge and research in dairy technologies and lactic acid bacteria (LAB) and dairy associated species in the fermentation of dairy products. The text deals with the industrial processing of milk, the problems solved in the industry, and those still affecting the processes. The authors explore culture methods and species selective growth media, to grow, separate, and characterize LAB and dairy associated species, molecular methods for species identification and strains characterization, Next Generation Sequencing for genome characterization, comparative genomics, phenotyping, and current applications in dairy and non-dairy productions.
In addition, Microbiology in Dairy Processing covers the Lactic Acid Bacteria and dairy associated species (the beneficial microorganisms used in food fermentation processes): culture methods, phenotyping, and proven applications in dairy and non-dairy productions. The text also reviews the potential future exploitation of the culture of novel strains with useful traits such as probiotics, fermentation of sugars, metabolites produced, bacteriocins. This important resource:
- Offers solutions both established and novel to the numerous challenges commonly encountered in the industrial processing of milk and the production of milk products
- Takes a highly practical approach, tackling the problems faced in the workplace by dairy technologists
- Covers the whole chain of dairy processing from milk collection and storage though processing and the production of various cheese types
Written for laboratory technicians and researchers, students learning the protocols for LAB isolation and characterisation, Microbiology in Dairy Processing is the authoritative reference for professionals and students.
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Table of Contents
Cover
Title Page
List of contributors
Foreword
Preface
Acknowledgements
1 Milk fat components and milk quality
1.1 INTRODUCTION
1.2 CONCLUSIONS
References
2 Spore‐forming bacteria in dairy products
2.1 INTRODUCTION
2.2 THE BACTERIAL SPORE
2.3 SPORE‐FORMING BACTERIA IMPORTANT FOR THE DAIRY INDUSTRY
2.4 CONTROL STRATEGIES TO PREVENT POISONING AND SPOILAGE OF MILK AND DAIRY PRODUCTS BY SPORE‐FORMING BACTERIA
2.5 CONCLUSIONS
References
3 Psychrotrophic bacteria
3.1 INTRODUCTION
3.2 SOURCES OF PSYCHROTROPHIC BACTERIA CONTAMINATION OF MILK
3.3 IMPORTANT SPOILAGE PSYCHROTROPHIC BACTERIA IN MILK
3.4 MOLECULAR TOOLS TO CHARACTERIZE PSYCHROTROPHIC BACTERIA
3.5 INFLUENCE OF PSYCHROTROPHIC CONTAMINATION OF RAW MILK ON DAIRY PRODUCT QUALITY
3.6 REGULATION OF EXTRACELLULAR ENZYMES
3.7 CONTROL OF PSYCHROTROPHIC BACTERIA AND RELATED ENZYMES
3.8 CONCLUSIONS
References
4 Stabilization of milk quality by heat treatments
4.1 INTRODUCTION
4.2 THERMAL TREATMENTS OF MILK
4.3 MILK STERILIZATION
4.4 DISEASES ASSOCIATED WITH UNPASTEURIZED MILK, OR POST‐PASTEURIZATION DAIRY‐PROCESSING CONTAMINATION
4.5 CONCLUSIONS
References
5 Genomics of LAB and dairy‐associated species
5.1 INTRODUCTION
5.2 GENOMICS OF LAB AND DAIRY‐ASSOCIATED SPECIES
5.3 NGS PLATFORM APPLIED TO SEQUENCING OF MICROBIAL COMMUNITIES
5.4 METABOLOMICS AND PROTEOMICS
5.5 COMPARATIVE GENOMICS OF DAIRY‐ASSOCIATED BACTERIA: THE
LACTOBACILLUS
GENUS COMPLEX,
STREPTOCOCCI/LACTOCOCCI
,
ENTEROCOCCI
,
PROPIONIBACTERIA
AND
BIFIDOBACTERIA
5.6 CLUSTERED REGULARLY INTERSPACED SHORT PALINDROMIC REPEATS (CRISPR) IN ADAPTIVE IMMUNITY
5.7 REGULATION IN CARBON METABOLISM
5.8 CONCLUSIONS
References
6 Metabolism and biochemistry of LAB and dairy‐associated species
6.1 INTRODUCTION
6.2 CARBOHYDRATE SUBSTRATES, GLYCOLYSIS AND ENERGY PRODUCTION
6.3 PROTEOLYSIS, PROTEIN SUBSTRATES AND AMINO ACID AVAILABILITY INFLUENCING GENE EXPRESSION
6.4 LIPOLYSIS, LIPASES, ESTERASES
6.5 AROMA AND FLAVOUR PRODUCTS OF METABOLISM
6.6 NONENZYMATIC PRODUCTION OF FLAVOURS
6.7 METHODS OF ANALYSIS OF FLAVOURS IN DAIRY PRODUCTS: HPLC, GAS CHROMATOGRAPHY/MASS ANALYSIS (GC/MS)
6.8 NATURAL BIODIVERSITY OF STRAINS IN DAIRY PRODUCTIONS
6.9 CONCLUSIONS
7 Growth needs and culture media for LAB and dairy‐associated species
7.1 INTRODUCTION
7.2 ESTABLISHED CULTURE MEDIA FOR LACTOBACILLI
7.3 M17 MEDIUM FOR SELECTION AND ENUMERATION OF LACTOCOCCI AND STREPTOCOCCI
7.4 SELECTIVE MEDIA FOR LACTOBACILLI
7.5 MEDIA FOR THE ISOLATION OF BIFIDOBACTERIA
7.6 PHENOTYPING
7.7 CONCLUSIONS
References
8 LAB species and strain identification
8.1 INTRODUCTION
8.2 GENOTYPIC FINGERPRINTING METHODS
8.3 CULTURE‐DEPENDENT APPROACHES
8.4 NON‐GENOTYPIC FINGERPRINTING METHODS
8.5 CULTURE‐INDEPENDENT APPROACHES
8.6 NOVEL HIGH‐THROUGHPUT TECHNIQUES: SEQUENCING AND METAGENOMICS
8.7 CONCLUSIONS
References
9 LAB strains with bacteriocin synthesis genes and their applications
9.1 INTRODUCTION
9.2 BACTERIOCINS FROM LAB
9.3 POTENTIAL FOR USE OF LAB BACTERIOCINS AS FOOD PRESERVATIVES
9.4 BACTERIOCINS PRODUCED BY DAIRY LAB
9.5 IDENTIFICATION OF LAB‐PRODUCING BACTERIOCINS
9.6 A NOVEL APPROACH FOR SCREENING LAB BACTERIOCINS
9.7 BIOTECHNOLOGICAL INTERVENTIONS FOR BACTERIOCIN ENGINEERING
9.8 CONCLUSIONS
References
10 Starter strains and adjunct non‐starter lactic acid bacteria (NSLAB) in dairy products
10.1 INTRODUCTION
10.2 CONTROLLED FERMENTATION
10.3 ADJUNCT NON‐STARTER LACTIC ACID BACTERIA
10.4 CONCLUSIONS
References
11 Milk fat: stability, separation and technological transformation
11.1 INTRODUCTION
11.2 PHYSICAL INSTABILITY OF MILK FAT
11.3 MILK FAT SEPARATION
11.4 PARTIAL COALESCENCE
11.5 FOAM IN MILK AND CREAM
11.6 WHIPPED CREAM AND BUTTER
11.7 CHURNING PROCESS
11.8 CONCLUSIONS
References
12 Biological traits of lactic acid bacteria: industrial relevance and new perspectives in dairy applications
12.1 INTRODUCTION
12.2 SELECTING FERMENTING BACTERIA FOR THEIR ABILITY TO HAVE A RESPIRATORY METABOLISM
12.3 SELECTING GALACTOSE‐POSITIVE YOGURT CULTURES: WORKING “AGAINST THE NATURAL EVOLUTION OF THE SPECIES”
12.4 ACCELERATING THE MILK ACIDIFICATION PROCESS BY SELECTING PROTEINASE‐POSITIVE STRAINS
12.5 ACCELERATING THE MILK ACIDIFICATION PROCESS BY SELECTING UREASE‐NEGATIVE
S. thermophilus
STRAINS
12.6 PROTECTIVE CULTURES FOR DAIRY APPLICATIONS: “WORK BUT PLEASE DO NOT GROW AND DO NOT MODIFY THE SENSORY PROFILE OF THE PRODUCT”
12.7 SELECTION OF STARTER CULTURE FREE OF TRANSFERABLE ANTIBIOTIC‐RESISTANCE MECHANISMS
12.8 CONCLUSIONS
References
13 Lactic acid bacteria bacteriophages in dairy products: problems and solutions
13.1 INTRODUCTION
13.2 PHAGE CLASSIFICATION
13.3 PHAGE‐HOST INTERACTIONS
13.4 SOURCES OF CONTAMINATION
13.5 PHAGE DETECTION AND QUANTIFICATION
13.6 METHODS TO CONTROL PHAGE CONTAMINATION
13.7 CONCLUSIONS
References
14 Lactic acid bacteria: a cell factory for delivering functional biomolecules in dairy products
14.1 INTRODUCTION
14.2 VITAMINS
14.3 MINERALS
14.4 BIOACTIVE COMPOUNDS
14.5 LOW‐CALORIE SWEETENERS
14.6 EXOPOLYSACCHARIDES (EPS)
14.7 CONCLUSIONS
References
15 Dairy technologies in yogurt production
15.1 INTRODUCTION
15.2 YOGURT TYPES
15.3 YOGURT MANUFACTURING PROCESS
15.4 CONCLUSIONS
References
16 Milk protein composition and sequence differences in milk and fermented dairy products affecting digestion and tolerance to dairy products
16.1 INTRODUCTION
16.2 CASEINS
16.3 PROTEOLYTIC RELEASE OF BIOACTIVE PEPTIDES IN FERMENTED MILK AND CHEESE
16.4 MINOR MILK PROTEINS
16.5 PROTEINS WITH BIOACTIVE ROLES
16.6 MFGM‐ASSOCIATED PROTEINS
16.7 COW’S MILK PROTEIN ALLERGY (CMPA)
16.8 CONCLUSIONS
References
Index
End User License Agreement
List of Tables
Chapter 01
Table 1.1 Average values in literature for fat content, milk fat globules characteristics and fatty acid composition of milk from different species.
Table 1.2 Calculated average values for fat content and some fatty acids (g/l) in milk from different species and Dietary Reference Values.
Chapter 02
Table 2.1 Strong‐evidence foodborne outbreaks caused by
Bacillus
and
Clostridium
toxins reported by the EFSA and the ECDC from 2010 to 2014.
Chapter 03
Table 3.1 Heat‐stability of proteases secreted by
Pseudomonas
genus.
Chapter 06
Table 6.1 LAB metabolism and type of fermentation.
Table 6.2 LAB Peptidases.
Table 6.3 Major volatile odor‐active compounds in dairy products.
Table 6.4 Descriptions of some important flavour components and their thresholds.
Table 6.5 Flavour compounds from amino acids by transamination and α‐keto acid formation.
Chapter 07
Table 7.1 Rogosa composition (per litre of purified water).
Table 7.2 MRS composition (per litre of purified water).
Table 7.3 M17 composition (per litre of purified water).
Table 7.4 ST agar composition (per litre of purified water) and comparison with Elliker broth.
Table 7.5 Lc agar composition (per litre of purified water).
Table 7.6 M‐RTLV composition.
Table 7.7 MMV agar composition.
Table 7.8 Homofermentative‐heterofermentative differential (HHD) medium.
Table 7.9 GAM broth (g/l).
Table 7.10 NNLP medium composition.
Table 7.11 MRS‐ABC medium. A, B and C solutions composition.
Chapter 09
Table 9.1 Classes of bacteriocins produced by Lactic Acid Bacteria (LAB).
Table 9.2 PCR references for the detection of bacteriocin gene clusters.
Table 9.3 Revised classification for bacteriocins of LAB.
Chapter 11
Table 11.1 Composition in saturated fatty acids of bovine milk fat (I =
iso
; AI =
anteiso
).
Table 11.2 Composition in unsaturated fatty acids of bovine milk fat.
Table 11.3 Characteristics of polymeric films related to the polymer properties (data from various sources).
Table 11.4 Interfacial tensions (γ) at pure substances contact and in some potential interface in milk.
Table 11.5 Gross composition of different types of milk cream.
Chapter 13
Table 13.1 Methods of bacteriophage detection.
Table 13.2 Phage‐control strategies in dairy plants.
Chapter 14
Table 14.1 Comparison of lactic acid bacteria, GABA production conditions and final content in milk or dairy products.
Chapter 15
Table 15.1 Composition of milk of selected species.
List of Illustrations
Chapter 02
Figure 2.1 Scheme of bacterial spore structure (layers are not drawn to scale). DNA: deoxyribonucleic acid; DPA: dipicolinic acid; SASPs: small acid‐soluble spore proteins.
Figure 2.2 Life cycle of spore‐forming bacteria. DPA: dipicolinic acid; SASPs: small acid‐soluble spore proteins.
Figure 2.3 Hard ovine milk cheese with LBD.
Chapter 04
Figure 4.1 Diagram of milk heating vs. time length. Degree of loss in nutritional value in lysine.
Chapter 08
Figure 8.1 Flowchart of the current molecular techniques used singularly or in combination to study dairy microbiota.
Chapter 09
Figure 9.1 Structure of some lantibiotic encoding operons and function of the respective gene products. Panel I: gene arrangement in the nisin (a) (Lubelski et al., 2008), lacticin 481 (b) (Rincé et al., 1997), and lacticin 3147 (c) gene clusters (McAuliffe et al., 2001). T: terminator is often represented with a circle on the stem. Panel II: function and cellular localization of the nisin modification, export and immunity proteins.
Figure 9.2 Structure of nisin A, a 3353‐Da cationic, linear peptide of 34 amino acids, containing five intramolecular ring structures.
Figure 9.3 Rapid screening system for novel LAB bacteriocins.
Chapter 11
Figure 11.1 Percentage of surface skimming of milk fat over time. The geometry of surfacing basin is suited to lower the time needed in respect to the technological performance.
Figure 11.2 Efficiency factor and percentage of fat content in separated milk as a function of separation temperature. The dashed line corresponds to the theoretical fat percentage in separated milk, according to the formula of Stokes.
Figure 11.3 Section of a centrifugal separator.
Figure 11.4 Conical stainless steel discs of centrifugal separator.
Figure 11.5 Grade of coalescence (number of events per time unit) as a function of mean globule diameter and of fraction of fat crystallized. (Approximate data from various sources.)
Figure 11.6 Representation of two fat globules with different sizes immediately before collision. The bars with circle distributed on the globule surfaces represent small‐size surfactant (P‐lipids, for instance).
Figure 11.7 Schematic representation of a casein micelle C adhering to a fat globule (D) immersed in the whey phase (S). (Arrows
γ
SC
,
γ
SD
and
γ
CD
are vectors corresponding to the three interface tensions).
Figure 11.8 Oil droplet (A) and milk fat globule (C and E) interaction with air cell‐milk plasma different conditions. Interfaces with adsorbed surface active substances are indicated by a thick line.
Figure 11.9 Foam‐stabilizing network realized by clumps of partially coalescent fat globules linked to air cells. The represented structure is that of whipped cream. A: air cell; G: clump of globules.
Figure 11.10 Schematic representation of fat globules and air cell during churning.
Figure 11.11 Example of thermal cycles to which a cream is subjected before churning.
Figure 11.12 General scheme of physical and biological cream maturation before churning.
Figure 11.13A Initial steps of cream preparation and treatment for churning process up to the attainment of butter granules.
Figure 11.13B Steps following churning.
Chapter 15
Figure 15.1 Flow chart of yogurt manufacturing process
Figure 15.2 Typical graph of yogurt fermentation, pH vs time and viscosity vs time
Figure 15.3 Simplified representation of the metabolic route in yogurt starter culture
Chapter 16
Figure 16.1 Differences in milk proteins, lactose and fats in milks of 15 species.
Guide
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Microbiology in Dairy Processing
Challenges and Opportunities
Edited by Palmiro Poltronieri
Institute of Sciences of Food Productions (CNR‐ISPA)
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Library of Congress Cataloging‐in‐Publication Data
Names: Poltronieri, Palmiro, editor.Title: Microbiology in dairy processing : challenges and opportunities / edited by Palmiro Poltronieri.Description: Hoboken, NJ : John Wiley & Sons, 2017. | Series: IFT Press series | Includes index. | Identifiers: LCCN 2017016747 (print) | LCCN 2017035565 (ebook) | ISBN 9781119114970 (pdf) | ISBN 9781119114987 (epub) | ISBN 9781119114802 (cloth)Subjects: LCSH: Dairy microbiology. | Dairy processing.Classification: LCC QR121 (ebook) | LCC QR121 .M54 2017 (print) | DDC 579.3/7–dc23LC record available at https://lccn.loc.gov/2017016747
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List of contributors
Iolanda AltomonteDepartment of Veterinary Sciences, University of Pisa, Pisa, Italy
Maria AponteDepartment of Agricultural Sciences, Division of Microbiology, University of Naples Federico II, Portici, Italy
Stefania ArioliDepartment of Food Environmental Nutritional Sciences (DeFENS), University of Milan, Milan, Italy
Marta Ávila ArribasDepartamento de Tecnología de Alimentos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain
François BaglinièreDepartment of Microbiology, Federal University of Viçosa, Minas Gerais, Brazil
Giovanna BattelliISPA‐CNR, Institute of Sciences of Food Productions, Milano, Italy
Giuseppe BlaiottaDepartment of Agricultural Sciences, Division of Grape and Wine Sciences, University of Naples Federico II, Avellino, Italy
Milena BrascaISPA‐CNR, Institute of Sciences of Food Productions, Milano, Italy
Cinzia CaggiaDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Catania, Italy
Cesare CammàIstituto Zooprofilattico Sperimentale ‘G. Caporale’, Teramo, Italy
Domenico CarminatiCREA‐ZA, Centro di ricerca Zootecnia e Acquacoltura, sede di Lodi, Italy
Laura CavallarinCNR‐ISPA, Institute of Sciences of Food Productions, Dipartimento di Scienze Veterinarie dell’Università di Torino, Grugliasco, TO, Italy
Luca Simone CocolinSettore di Microbiologia agraria e Tecnologie alimentari, Dipartimento di Scienze Agrarie, Forestali e Alimentari, Grugliasco, Italy
Fabio Dal BelloSacco System, Sacco srl, Cadonago (CO), Italy
Maria Cristina Dantas VanettiDepartment of Microbiology, Federal University of Viçosa, Minas Gerais, Brazil
Marilù DecimoISPA‐CNR, Institute of Sciences of Food Productions, Milano, Italy
Elisabetta Di GiannataleIstituto Zooprofilattico Sperimentale, “G. Caporale”, Teramo, Italy
Paola DolciSettore di Microbiologia agraria e Tecnologie alimentari, Dipartimento di Scienze Agrarie, Forestali e Alimentari, Grugliasco, Italy
Sonia Garde Lopez‐BreaDepartamento de Tecnología de Alimentos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain
Giorgio GiraffaCREA‐ZA, Centro di ricerca Zootecnia e Acquacoltura, sede di Lodi, Italy
Marzia GiribaldiCREA‐IT, Research Centre for Engineering and Agro‐Food Processing, Turin, Italy
Maria Gabriella GiuffridaCNR‐ISPA, Institute of Sciences of Food Productions, Dipartimento di Scienze Veterinarie dell’Università di Torino, Grugliasco, TO, Italy
Natalia Gómez‐TorresDepartamento de Tecnología de Alimentos, Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA), Madrid, Spain
Solimar Gonçalves MachadoFederal Institute of Education, Science and Technology of North of Minas Gerais, Campus Salinas, Minas Gerais, Brazil
Nicoletta Pasqualina MangiaDepartment of Agriculture, University of Sassari, Italy
Mina MartiniDepartment of Veterinary Sciences, University of Pisa, Pisa, Italy
Giacomo MiglioratiIstituto Zooprofilattico Sperimentale, “G. Caporale”, Teramo, Italy
Diego MoraDepartment of Food Environmental Nutritional Sciences (DeFENS), University of Milan, Milano, Italy
Stefano MorandiISPA‐CNR, Institute of Sciences of Food Productions, Milano, Italy
Alessandra PinoDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Catania, Italy
Palmiro PoltronieriInstitute of Sciences of food Productions (CNR‐ISPA), CNR, National Research Council of Italy, Lecce, Italy
Francesco PomilioIstituto Zooprofilattico Sperimentale, “G. Caporale”, Teramo, Italy
Cinzia RandazzoDepartment of Agriculture, Food and Environment (Di3A), University of Catania, Catania, Italy
Franca RossiIstituto Zooprofilattico Sperimentale, “G. Caporale”, Teramo, Italy
Lorena SacchiniIstituto Zooprofilattico Sperimentale, “G. Caporale”, Teramo, Italy
Tiziana SilvettiISPA‐CNR, Institute of Sciences of Food Productions, Milano, Italy
Gianluigi ScolariIstituto di Microbiologia degli alimenti, Università Cattolica del Sacro Cuore, Piacenza, Italy
Panagiotis SfakianakisFood Chemistry and Technology, School of Chemical Engineering, National Technical University of Athens, Zografou, Greece
Federica SalariDepartment of Veterinary Sciences, University of Pisa, Pisa, Italy
Constantina TziaFood Chemistry and Technology, School of Chemical Engineering, National Technical University of Athens, Zografou, Greece
Koenraad van HoordeLaboratory of Brewing and Biochemistry, Faculty of Bioscience Engineering, Ghent University, Gent, Belgium
Miriam ZagoCREA‐ZA, Centro di ricerca Zootecnia e Acquacoltura, sede di Lodi, Italy
Foreword
Microbiology in Dairy Processing: Challenges and Opportunities is directed to the following: dairy scientists; dairy professionals in industry and academia; those in food science, dairy science and microbiology; intermediate course and post‐graduate students; trained laboratory personnel; and R&D and production personnel in dairy industry companies of all sizes. The idea to write this book came from the section “Questions” in the Researchgate community. I realised that there is a need to introduce lactic acid bacteria (LAB) growth media at various levels of expertise, from young researchers starting their laboratory work to food technologists devoted to microbiological analyses. Therefore, from this starting point, I searched the recent literature and produced a list of exceptionally interesting publications on how far the genomics field has advanced in its knowledge of LAB species in recent years. The chapters in this book reflect these advancements and offer a panoramic view of the research fields in which to apply these advancements in knowledge, either for LAB and dairy‐associated species and their applications in dairy productions and for the technologies to maintain the milk products safe and devoid of undesired pathogens and milk spoilage bacteria. The challenges of dairy microbiology are either to maintain the product safety devoid of undesired bacteria that may spoil the quality and change the taste or to the further advancement in the microbiota and the interaction among bacteria at community level. The opportunities remain in the exploration of the biodiversity of LAB and dairy‐associated species, either at genome rearrangements and horizontal gene transfer or at the biochemistry level, to produce novel dairy products that are low fat, low salt, or with beneficial properties for human health.
Preface
Microbiology in Dairy Processing: Challenges and Opportunities introduces and reviews the knowledge regarding dairy technologies and lactic acid bacteria (LAB) and dairy‐associated species in the fermentation of dairy products for laboratory technicians and researchers and students learning the protocols for LAB isolation and characterisation. It provides application notes useful in laboratories of food technology departments and for students and researchers studying all aspects of the milk‐processing industry, from microbiology to food productions.
The chapters deal with the industrial processing of milk – the problems solved and those still affecting the processes, from microfiltration to deterioration of stored milk in cold by psychrotrophic bacteria (such as Pseudomonas fragi) and by spore‐forming bacteria – and cheese‐manufacturing technologies. The book introduces culture methods and species‐selective growth media to grow, separate and characterise LAB and dairy‐associated species, molecular methods for species identification and strain characterization, Next Generation Sequencing for genome characterization, comparative genomics, phenotyping, and current applications in dairy and non‐dairy productions, as well as the potential future exploitation of the culture of novel strains with useful traits (probiotics, fermentation of sugars, metabolites produced, bacteriocins).
Chapter 1 introduces the quality and properties of milk fats and differences in milks of various origin. Chapter 2 overviews the spore‐forming bacteria associated with milk. Chapter 3 discusses the problem of psychrotrophic bacteria in milk deterioration. Chapter 4 presents the various types of industrial milk according to the freshness and quality. Chapter 5 presents the advancements in LAB and dairy‐associated species genomics and strain differences, related to gene content and their applications. Chapter 6 presents very broadly the biochemistry of LAB and dairy‐associated species. Chapter 7 reviews selective growth media for different species of LAB and non‐LAB dairy‐associated bacteria. Chapter 8 introduces the molecular tools for strain identification and characterization. Chapter 9 discusses the bacteriocin‐producing LAB species and their potential applications in food products. Chapter 10 analyses in detail the complex interactions among starter and non‐starter strains. Chapter 11 reviews the physical‐chemical properties of milk cream products and technological processes involving milk fats and cream‐derived products. Chapter 12 analyses technological traits of lactic acid bacteria, their industrial relevance and new perspectives. Chapter 13 overviews LAB bacteriophages in dairy products, their problems and solutions. Chapter 14 details the application of LAB as a cell factory for delivering functional biomolecules in dairy products. Chapter 15 reviews the dairy technologies applied to yogurt production. Finally, Chapter 16 introduces properties of milk proteins, the differences in amino acids of protein variants, and the potential to originate bioactive peptides and the proteolysis by co‐fermenting LAB species, a process that may ensure the safety and healthiness of the fermented products, as assessed by EFSA authority. Last, the potential for milks of different origin to be administered to individuals suffering of milk allergies or intolerance is discussed.
Palmiro PoltronieriInstitute of Sciences of Foods Productions (CNR‐ISPA)
Acknowledgements
The contributions of all members of staff at Wiley–IFT who were involved with writing and reviewing the draft of this book are thankfully acknowledged. This book was made possible thanks to the support of colleagues and professors from Italy, Spain, Belgium, Greece, and Brazil. Among others, I would like to thank Dr. Bruno Battistotti, whose course in dairy technologies opened my mind and my working perspectives during my first years of research; Dr. Pier Sandro Cocconcelli, who introduced me to LAB molecular methods; Drs. Giuseppe Zacheo, Franco Dellaglio and Marco Gobbetti, for supervising microbial technologies and project proposals of my Institute; and Dr. Maria Morea for including me in her projects on “Microbiology for food quality and safety”. A special appreciation goes to the contributors of the book chapters and to AITeL, the Italian Association of Milk Technologists, providing the opportunity to meet and include some of the last‐minute contributors. Finally, I would like to thank the scientific community, the European Research Council (previously ESF), for the opportunity to work and collaborate with high‐level scientists in several international projects, such as Ferbev, Riboreg, TransBio, and COST 853 “Agricultural biomarkers for array technology”. This book was made possible thanks to the scientific vision of many other colleagues who, because of their schedule, could not be directly involved at this time, but whose work has enlightened the process of writing the chapters and the organization of the book.
1Milk fat components and milk quality
Iolanda Altomonte, Federica Salari and Mina Martini
Department of Veterinary Sciences, University of Pisa, Pisa, Italy
1.1 INTRODUCTION
From a physico‐chemical point of view, milk is an emulsion of lipid globules and a colloidal suspension of protein and mineral aggregates in a solution of carbohydrates (mainly lactose). In Western countries, milk and dairy products, and in general food of animal origin, are often accused of causing adverse health effects, especially with regard their food lipid intake, since lipids have been implicated in several diseases such as obesity, insulin resistance and atherosclerosis (Olofsson et al., 2009). For these reasons, the number of studies on the physical and chemical structure of fat in several edible products of animal origin have increased. Although milk and dairy products contain saturated fatty acids, they also provide specific beneficial components for human health and also lipid components (phospholipids, some individual fatty acids (FAs) and fat‐soluble vitamins) that have a role in health maintenance. In addition, milk is a major source of dietary energy, especially in developing countries, where there is shortage of animal‐source food (FAO, 2013), and in childhood.
Milks of different origins have long been used, and they have been processed to dairy products for their longer shelf life. Due to the wide natural variability from species to species in the proportion of milk macronutrients and to variations along lactation, milk represents a flexible source of nutrients that may be exploited to produce a variety of dairy products.
Ruminant milk is the main source available for humans to use to manufacture dairy products and fermented milk. Besides cow’s milk and milk from other ruminants (such as buffalo, goat and sheep), research on milk from other species is still poorly exploited (FAO, 2013). More recently, equine milks have been suggested for use in children with severe IgE‐mediated cow milk protein allergy (CMPA) (Monti et al., 2007, 2012; Sarti et al., 2016), and local producers have established a niche for the application of donkey products with well‐characterised profile of its constituents (Martini et al., 2014a).
1.1.1 Milk fat globules
Milk lipids are composed of milk fat globules (MFGs) made up of triglycerides enveloped by a biological membrane. MFGs are responsible and/or contribute to some properties and phenomena in milk and dairy products and may affect milk fatty acid composition and the way in which fat is digested (Baars et al., 2016; Huppertz and Kelly, 2006; Martini et al., 2017). For the dairy industry it is of interest that changes in the morphometry of the MFGs lead to changes in milk quality, yields, and ripening and the nutritional quality of cheeses (Martini et al., 2004).
In milk of different species there are MFGs of various sizes, ranging from a diameter smaller than 0.2 µm to a maximum of about 15 µm, with an average diameter that varies as a function of endogenous (species, breed), physiological (parity, stage of lactation), and exogenous factors (feeding) (Martini et al., 2010a).
Different average diameters have been reported in the literature for ruminant species (3.5–5.5 µm for cows; 2.79–4.95 µm for sheep; 2.2 and 2.5–2.8 µm for goats and 2.96–5.0 µm for buffalos) (Table 1.1) (Martini et al., 2016b). However average diameter of globules in equids is considerably lower than other dairy species (about 2 µm in donkey) (Martini et al., 2014b), while regarding human MFGs, larger dimensions have also been found (4 µm) (Lopez and Ménard, 2011).
Table 1.1 Average values in literature for fat content, milk fat globules characteristics and fatty acid composition of milk from different species.
Cow
Buffalo
Goat
Sheep
Donkey
Horse
Human
Fat
%
3.70
8.14
3.90
6.50
0.36
1.48
3.34
Average diameter of the fat globules
µm
3.5–5.5
2.96–5.0
2.2–2.8
2.79–4.95
2
2–3
3.3
SFA
g/100g fat
71.24
65.9
70.42
71.85
55.55
45.18
41.77
MUFA
g/100g fat
25.56
31.4
25.67
26.04
22.21
31.88
38.73
PUFA
g/100g fat
3.20
2.70
4.08
2.10
21.08
22.93
16.96
UFA
g/100g fat
28.76
34.1
29.75
28.14
43.29
54.81
55.29
UFA:SFA ratio
0.40
0.52
0.42
0.39
0.78
1.20
1.32
SCFA
g/100g fat
10.52
9.72
17.51
17.13
12.29
10.79
1.87
MCFA
g/100g fat
52.81
53.70
48.28
45.87
40.08
42.47
37.94
LCFA
g/100g fat
34.38
32.73
32.64
35.87
47.64
46.75
57.72
CLA c9, t11
g/100g fat
0.65
0.45
0.70
1.00
–
0.09
0.19
C18:2 n6 (LA)
g/100g fat
2.42
1.71
2.72
1.20
9.5
16.17
12.96
C18:3 n3 (ALA)
g/100g fat
0.25
0.51
0.53
0.77
7.25
5.96
1.15
C18:2 n6: C18:3 n3 ratio
––
9.68
3.35
5.13
1.56
1.31
2.71
11.26
C20:4 (AA)
g/100g fat
0.13
0.10
0.16
0.10
0.09
0.10
0.4
C20:5 (EPA)
g/100g fat
0.05
0.03
nd
nd
0.26
–
0.11
C22:6 (DHA)
g/100g fat
nd
–
0.05
0.04
0.28
–
0.51
SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids; UFA: unsaturated fatty acids; SCFA: short chain fatty acids (≤10C); MCFA: medium chain fatty acids (≤11C, ≥17C); LCFA: long chain fatty acids (≥18C); nd: no data.
The MFG membrane (MFGM) is a triple membrane resulting from the mammary secretory cell that surrounds a core of triglycerides distributed in a lamellar way (Heid and Keenan, 2005).
The MFGM consists of different classes of lipids (phospholipids, triglycerides and cholesterol) and of several proteins and enzymes. Phospholipids, in the form of mixtures of fatty acid esters of glycerol and sphingosine, possibly containing phosphoric acid, and a nitrogen‐based compound (choline, ethanolamine or serine). These are natural emulsifiers able to maintain the milk lipids as discrete globules, ensuring high stability. MFGM contains about 1% of the total milk proteins. Most of them are present in very low amounts and are enzymes and proteins involved in milk synthesis. The principal proteins in the MFGM include mucins (MUC) 1 and 5, adipophilin (ADPH), butyrophilin (BTN), periodic acid‐Schiff glycoproteins (PAS) 6 and 7, fatty acid binding protein (FABP), and xanthine oxidoreductase (XOR), a metal (Mo, Fe) binding protein (Spertino et al., 2012). In the last few years, research on the composition and structure of the milk membranes have been increased and have improved the knowledge of the MFGM from species other than the bovine (Saadaoui et al., 2013; Pisanu et al., 2012; Lu et al., 2016; Martini et al., 2013).
These studies have increased also due to the fact that MFGM is a dietary source of functional substances and is considered a nutraceutical (Rosqvist et al., 2014; Timby et al., 2015; Hernell et al., 2016). The functionality of the MFGM seems to be provided by its content of phospholipids, sphingolipids, fatty acids and proteins with an antibacterial effect (such as xanthine oxidoreductase and mucins) and/or health benefits.
MFGM conveys fat in an aqueous environment and is damaged by some treatment, such as homogenization, whipping and freezing, affecting milk physicochemical properties, for example producing hydrolytic activity, rancidity, and oiling off, and low wettability of milk powders. MFGM composition also affects the creaming rate on the milk surface (Martini et al., 2017); in bovine milk this phenomenon is due to the effect of cryoglobulins, an M‐type immunoglobulin that aggregates globules during cold storage. Other types of milk are lacking these cryoglobulins and do not agglutinate. Homogenization reduces globule diameter, making globules insensitive to the action of cryoglobulins and prevents agglutination. During butter production, extensive agitation and kneading causes the MFGM to form the water‐in‐oil emulsion. The partitioning in the aqueous phase produces the loss of MFGM in the buttermilk.
1.1.2 Milk fat and fatty acid composition
Milk lipid content and fatty acid composition vary by virtue of various endogenous and exogenous factors. Among endogenous factors, the species, breed and stage of lactation are the main factors.
Regarding the species, buffalo and sheep milk contains higher fat percentages and are particularly suitable for processing, such as cheese making. Fat percentages vary in a range between 7 and 9% for buffalo, but can reach 15% under favourable conditions (Altomonte et al., 2013; Varricchio et al., 2007), whereas in sheep the range is between 6.5 and 9% depending on the breed (Haenlein, 2007; Martini et al., 2012). Regarding cow and goat milk, fat content are comparable; in fact cow total lipid ranges from 3.4% in Holstein to about 6% in Jersey breeds (Nantapo et al., 2014; Pegolo et al., 2016; Sanz Ceballos et al., 2009), and goat range from a minimum of 3.5% to a maximum of 5.6% in some native goats (Haenlein, 2007; Martini et al., 2010b). Equid milk has lower fat percentages compared to ruminant milk; the average values reported in literature are 0.30–0.53% in donkey and 1.5% in horse milk (Pikul and Wójtowski, 2008; Martemucci and D’Alessandro, 2012; Martini et al., 2014b; Salimei et al., 2004). Furthermore, some authors stated lower contents (1.04%, 0.92%, 0.8%) in the milk of Halfinger, Hucul and Wielkopolski mares, respectively (Salamon et al., 2009; Pieszka Huszczyński and Szeptalin, 2011). The low fat content in equid milk could be a limiting factor in its use in infant nutrition in a diet exclusively based on milk, thus an appropriate lipid integration should be introduced. On the other hand it is encouraging for studies on the possible use of donkey milk in dietotherapy.
Regarding human milk, fat content is more similar to cow milk, varying between 2.8 and 3.8% (Antonakou et al., 2013).
From a nutritional point of view, donkey milk leads to lower saturated fatty acid (SFA) intake, about 2.00 g/l (Table 1.2), than the other milks commonly used for human feeding. Despite being rich in unsaturated fatty acids (UFAs) and having a UFA:SFA ratio intermediate between ruminant and human milk, donkey provides a limited amount of fat; thus, the total intake of UFA per 1 l of milk is lower (1.56 g) than milk of other species (Martini et al., 2016a).
Table 1.2 Calculated average values for fat content and some fatty acids (g/l) in milk from different species and Dietary Reference Values.
Source: Data from EFSA (2010).
Dietary Reference Values
Cow
Buffalo
Goat
Sheep
Donkey
Horse
Human
Fat
g/l
Adults: 20–30% of the energy of the diet (E); Infants (6–12 months): % 40 E%; Children (2–3 years): 35–40 E%.
37.0
81.4
39.0
65.0
3.60
14.8
33.4
SFA
g/l
as low as possible
26.36
53.64
27.46
46.70
2.00
6.68
13.95
MUFA
g/l
Not set
9.45
25.56
10.01
16.93
0.80
4.71
12.94
PUFA
g/l
Not set
1.18
2.20
1.59
1.36
0.76
3.39
5.66
UFA
g/l
Not set
10.63
27.76
11.60
18.29
1.56
8.1
18.60
C18:2 n6 (LA)
g/l
Adequate Intake (AI): 4 E%
0.89
1.39
1.06
0.78
0.34
2.39
4.33
C18:3 n3 (ALA)
g/l
AI: 0.5% E
0.09
0.41
0.21
0.50
0.26
0.88
0.38
C20:4 (AA)
g/l
Not set
0.05
0.08
0.06
0.06
0.006
0.01
0.03
C20:5 (EPA)
g/l
Not set
0.02
0.02
–
nd
0.009
–
0.04
C22:6 (DHA)
g/l
Infants and young children (between 6 and 24 months) AI: 0.10 g DHA
–
–
0.02 (20% of AI)
0.03 (30% of AI)
0.010 (9% of AI)
–
0.17 (170.34% of AI)
C20:5 + C22:6 (EPA + DHA)
g/l
Adults. AI: 0.25 g
–
–
–
–
0.017 (6.80% AI)
–
0.21 (84% of AI)
SFA: saturated fatty acids; MUFA: monounsaturated fatty acids; PUFA: polyunsaturated fatty acids; UFA: unsaturated fatty acids; nd: no data.
In milk from ruminants, especially sheep and goats, triglycerides contain short chain fatty acids (SCFAs) such as butyric acid and hexanoic, octanoic and decanoic acid. On the contrary, human (Yuhas, Pramuk and Lien, 2006) and donkey milk (Martini et al., 2014b) are characterized by low amounts of SCFA—especially the chains shorter than C8—and high quantities of long chain fatty acids (LCFAs).
SCFAs are synthesized by the fermentation of dietary fibre, are water soluble and volatile, and contribute to the typical flavour of ovine and caprine milk. When freed by endogenous lipase or bacterial enzymes, SCFA can also give rancidity and quality deterioration. SCFAs and MCFAs, which are a source of rapidly available energy, are particularly relevant for people suffering from malnutrition or fat absorption syndromes and for elderly people (Raynal‐Ljutovac et al., 2008).
Recent studies have highlighted effects of SCFA at cellular and molecular levels in the organism; their presence or their deficiency may affect pathogenesis of some diseases (autoimmune, inflammatory diseases). In addition, SCFAs have antimicrobial activity and anti‐inflammatory effects in the gut (Tan et al., 2014).
Ruminant milk, in particular milk from sheep that feed in pastures, is the richest natural source of conjugated linoleic acids (CLA) and of C18:1 trans‐11 (vaccenic acid) (Bauman and Lock, 2005; Lim et al., 2014). The CLA content in milk varies depending on species, breed and individual, farming system, feeding and season. In sheep milk CLA varies from 1.2 to 2.9 g/100 g of fat; in goat between 0.5 and 1 g/100 g of fat (Parodi, 2003). Cow milk is generally reported to vary from 0.1 to 2.2 g/100 g total FA (Elgersma, Tamminga and Ellen, 2006), whereas human and equid milk are poor sources of CLA (Table 1.1).
Ninety percent of CLA isomers in milk is made up of cis‐9, trans‐11C18:2 (rumenic acid) produced mainly by stearoyl Co‐A desaturase (SCD) o‐∆9–desaturase enzyme in the mammary gland using vaccenic acid as precursor, but also by the rumen bacterium Butyrivibrio fibrisolvens as intermediate of biohydrogenation of linoleic and linolenic acids ingested with feed (Bauman and Lock, 2005). Rumenic acid vary between 0.29 and 0.71% of total human milk fatty acids, while in the horse it is between 0.07 and 0.10%. Moreover, in equids, cecum seems to contribute little to CLA synthesis (Markiewicz‐Keszycka et al., 2014).
Anticarcinogenic properties and modulation of immunological functions have been demonstrated for rumenic acid in animal models and cell cultures (Field and Schley, 2004; O’Shea et al., 2004). However, the most documented effects of CLA in humans are the gain of muscle mass at the expense of body fat, whereas in vivo studies on the effects on atherosclerosis and cholesterol have shown conflicting results in humans (Crumb, 2011).
Vaccenic acid has shown anticancer properties in human mammary adenocarcinoma cells (Lim et al., 2014).
Regarding the omega‐3 FAs, milk is not a good source of this family of FAs. However, among the mammalian species reared for milk production, horse, sheep and donkey are richest sources of C18:3 n3 (α‐linolenic acid (ALA))(g/l) (Table 1.2), in particular donkey and horse milk provide a good ALA intake (0.22–0.88 g/l) although they have low fat content. In adults minimum intake levels for ALA are recommended to prevent deficiency symptoms (0.5% of energy) (FAO‐WHO, 2010).
Linoleic acid (LA) and ALA are precursors of omega 6 and omega‐3 families, respectively, and their ratio is generally considered as indicative of their balanced intake in the diet. The interest in the LA:ALA ratio derives from the antagonistic effects between the two families of FAs observed in human body. In fact, the higher intake of n‐6 fatty acids may reduce the formation of anti‐inflammatory mediators from omega‐3 fatty acids. Observations on animal models suggest that raising the n‐6 to n‐3 fatty acids ratio (n6:n3) acts on adipogenesis and the risk of obesity in the offspring later in life (Rudolph et al., 2015). However, research is yet not supported by studies in humans, and an optimal ratio of these fatty acids in the diet has not yet been established (EFSA, 2010). Furthermore, the prevalence of n‐6 in human diets has increased over the decades while n‐3 fatty acids remain unchanged, thus increasing the n‐6/n‐3 milk fatty acid ratio (Rudolph et al., 2015). Thus, a reduction of omega‐6 in the diet is desirable, and donkey’s milk appears to have a balanced rapport of these two families (about 1) compared to other milks (Martini et al., 2014b).
Arachidonic acid (AA) C20:4 is essential component of cellular membranes and also of MFGM, where it may have an essential role (Fong et al., 2007; Martini et al., 2013).
AA is present in almost similar amounts in the milk of ruminants (Table 1.2), while it shows lower values in equids.
Despite the importance of AA for membrane integrity (Fong, Norris and MacGibbon, et al., 2007), it has been described as an adipogenetic‐, pro‐inflammatory‐ and hypertension‐promoting factor (Vannice and Rasmussen, 2014), and recommended intake levels have not been established.
C20:5 (EPA) and C22:6 (DHA) have showed evidence of both independent and shared effects in neuroprotection and in the treatment for a variety of neurodegenerative and neurological disorders. In particular, DHA is an important constituent of the retina and the nervous system, and it has unique and indispensable roles in neuronal membranes (Dyall, 2015).
There is still insufficient evidence to support beneficial effects of EPA and DHA in foetal life or early childhood on obesity, blood pressure, or blood lipids (Voortman et al., 2015).
Overall levels of DHA and EPA in milk are quite low, and in human milk DHA content is highly variable; values from 0.17 to 0.99 % have been reported, depending on the diets and on different countries (Yuhas, Pramuk and Lien, 2006). The recommended daily intake of EPA plus DHA is 0.25 g in adults (EFSA, 2010).
1.2 CONCLUSIONS
The transformation of milks of different origin may be the source of dairy products with different and peculiar characteristics. Since a role in health maintenance has been reported for several lipid components of milk, a deep knowledge of milk lipid constituents from different dairy species is of utmost relevance for both the nutritional uptake and effects on human health.
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