Handbook of Probiotics and Prebiotics, 2nd Edition

Preface xv

Contributors xvii

Part I Probiotics 1

1 Probiotic Microorganisms 3

1.1 Definitions 3

1.2 Screening, Identification, and Characterization of Lactobacillus and Bifidobacterium Strains 4

1.2.1 Sources of Screening for Probiotic Strains 5

1.2.2 Identification, Classification, and Typing of Bifidobacterium Strains 7

1.2.2.1 Taxonomy 7

1.2.2.2 Identification and Typing 8

1.2.3 Identification, Classification, and Typing of Lactobacillus Strains 14

1.2.3.1 Taxonomy 14

1.2.3.2 Identification and Typing 15

1.2.4 Characterization of Probiotic Properties in Bifidobacterium and Lactobacillus Strains 19

1.2.4.1 Survival to GIT Stressing Conditions 19

1.2.4.2 Adhesion/Colonization to/of GIT 23

1.2.4.3 Antimicrobial Activity 24

1.2.4.4 Other Probiotic Properties 24

1.2.5 Conclusion 24

1.3 Detection and Enumeration of Gastrointestinal Microorganisms 25

1.3.1 Methods for Intestinal Microbiota Assessment 25

1.3.1.1 Culture-Dependent Methods 25

1.3.1.2 Culture-Independent Methods 30

1.3.2 Detection and Enumeration in Dairy Products 37

1.3.3 Detection and Enumeration of Specific Probiotics in the Gut 38

1.3.4 The Problem of the Viability and Physiological State of Intestinal Bacteria 41

1.3.5 Conclusions 42

1.4 Enteric Microbial Community Profiling in Gastrointestinal Tract by Terminal-Restriction Fragment Length Polymorphism (T-RFLP) 43

1.4.1 T-RFLP 43

1.4.2 Universal and Group-Specific Primers 44

1.4.3 Fluorescent Dyes 44

1.4.4 DNA Extraction 46

1.4.5 PCR Amplification 46

1.4.6 Generation of Terminal Restriction Fragments (TRF) by Digestion of Amplicons with Restriction Enzymes 46

1.4.7 Software and Data Processing 47

1.4.8 Microbial Diversity in Different Intestinal Compartments of Pigs 47

1.4.9 Tracking the Fate of Orally Delivered Probiotics in Feces 48

1.4.10 Conclusion 51

1.5 Effective Dosage for Probiotic Effects 52

1.5.1 Acute (Rotavirus) Diarrhea in Children 53

1.5.2 Antibiotic-Associated Diarrhea 54

1.5.2.1 Combination of L. acidophilus + bifidobacteria or Streptococcus thermophilus 54

1.5.2.2 L. rhamnosus GG or Saccharomyces boulardii Applied Singly 57

1.5.3 Helicobacter pyroli 58

1.6 Incorporating Probiotics into Foods 58

1.6.1 Probiotic Ingredients 60

1.6.2 Factors Affecting the Viability of Probiotics in Foods 60

1.6.2.1 Choice of Probiotic Organism/Food Combinations 61

1.6.2.2 Physiologic State of the Probiotic 63

1.6.2.3 Temperature 63

1.6.2.4 pH 64

1.6.2.5 Water Activity 64

1.6.2.6 Oxygen 65

1.6.2.7 Toxicity of Ingredients 66

1.6.2.8 Growth Factors, Protective, and Synergistic Ingredients 67

1.6.2.9 Freeze–Thawing 67

1.6.2.10 Sheer Forces 67

1.6.3 Synbiotics 67

1.6.4 Delivery Systems 68

1.6.4.1 Microencapsulation 68

1.6.4.2 Delivery Devices 69

1.6.5 Probiotic Foods 69

1.6.6 Conclusions 69

1.7 Safety of Probiotic Organisms 75

1.7.1 Current Proposals for Probiotic Safety 77

1.7.2 Taxonomic Identification 79

1.7.3 Pathogenicity 81

1.7.4 Antibiotic Resistance and Susceptibility 83

1.7.5 Immune Modulation 87

1.7.6 Clinical Studies 90

1.7.7 Postmarket Surveillance 92

1.7.8 GMO Probiotics 93

1.7.9 Conclusion 94

1.8 Legal Status and Regulatory Issues 95

1.8.1 Human Probiotics 95

1.8.1.1 Asia 95

1.8.1.2 Europe 106

1.8.1.3 The United States of America 111

1.8.2 Animal Probiotics 123

1.8.2.1 United States 123

1.8.2.2 European Union 123

1.8.2.3 China 125

1.8.2.4 Japan 125

1.8.2.5 Korea 125

1.8.2.6 Thailand 125

1.8.2.7 Australia 125

1.8.2.8 New Zealand 135

1.8.2.9 Indonesia, Malaysia, Philippines, and Vietnam 139

References 139

2 Selection and Maintenance of Probiotic Microorganisms 177

2.1 Isolation of Probiotic Microorganisms 177

2.2 Selection of Probiotic Microorganisms 178

2.2.1 Manufacturing Criteria (General Criteria) 179

2.2.2 Shelf Life and Gut Transit (General Criteria) 179

2.2.2.1 Shelf Life of Viable Probiotics Under Different Storage Conditions 179

2.2.2.2 Tolerance to Digestive Juices 180

2.2.2.3 Adhesion and Colonization onto Specific Site of Body Surface 181

2.2.3 Health Properties (Specific Criteria) 181

2.2.4 Safety 182

2.2.5 Identification 182

2.3 Maintenance of Probiotic Microorganisms 184

References 187

3 Genetic Modification of Probiotic Microorganisms 189

3.1 Mutants Obtained from Probiotic Microorganisms by Random Mutagenesis 189

3.2 Plasmids 202

3.3 Vectors for Lactobacilli and Bifidobacteria 212

3.4 Genetic Recombination 222

References 229

4 Role of Probiotics in Health and Diseases 257

4.1 Cell Line Models in Research 259

4.2 Laboratory Animal Models in Research 263

4.3 Effects on Human Health and Diseases 267

4.3.1 Nutritional Effects 267

4.3.1.1 Lactose Maldigestion 268

4.3.1.2 b-Galactosidase in Fermented Milk Products 269

4.3.2 Prevention and Treatment of Oral Infection and Dental Caries 270

4.3.3 Prevention and Treatment of Diarrhea 272

4.3.3.1 Acute (Rotavirus) Diarrhea in Children 272

4.3.3.2 Antibiotic-Associated Diarrhea 276

4.3.3.3 Clostridium difficile Associated Diarrhea 279

4.3.3.4 Radiation-Induced Diarrhea 279

4.3.3.5 Traveler’s Diarrhea 280

4.3.3.6 Diarrhea in Tube-Fed Patients 281

4.3.4 Treatment of Irritable Bowel Syndrome 282

4.3.5 Prevention and Treatment of Inflammatory Bowel Diseases 287

4.3.6 Treatment of H. pylori Infection 292

4.3.7 Prevention of Postoperative Infections 295

4.3.8 Prevention and Treatment of Respiratory Tract Infections 299

4.3.9 Prevention and Treatment of Allergic Diseases 302

4.3.10 Antitumor Effects 310

4.3.11 Reduction of Serum Cholesterol 313

4.3.12 Enhancement of Vaccine Responses 318

4.4 Effects on Farm Animals 321

4.4.1 Poultry 322

4.4.2 Swine 323

4.4.3 Ruminants 331

4.4.4 Rabbits 339

4.4.5 Pets 339

References 350

5 Mechanisms of Probiotics 377

5.1 Adhesion to Intestinal Mucus and Epithelium by Probiotics 377

5.1.1 Adhesion to Gastrointestinal Epithelial Cell Lines 378

5.1.2 Adhesion to Intestinal Mucus 378

5.1.3 Colonization of Probiotics in Human Intestine as Assessed by Biopsies 379

5.1.4 Comparisons Between In Vitro and In Vivo Results 379

5.1.5 Adhesins 379

5.1.6 Factors Affecting the Adhesion Properties of Probiotics 379

5.1.7 Adhesive and Inhibitory Properties of Nonviable Probiotics 380

5.1.8 Role of Age and Diseases on Adhesion 383

5.2 Combined Probiotics and Pathogen Adhesion and Aggregation 384

5.2.1 Aggregation 385

5.2.2 Adhesion 385

5.2.3 Assay for Adhesion 386

5.2.4 Assay for Aggregation 386

5.2.5 Factors that Determine Adhesion 389

5.2.6 In Vitro Models 389

5.2.7 Probiotics in Combination 390

5.2.8 Conclusion 391

5.3 Production of Antimicrobial Substances 391

5.3.1 Organic Acids 392

5.3.2 Hydrogen Peroxide 392

5.3.3 Carbon Dioxide 393

5.3.4 Bacteriocins 393

5.3.5 Low Molecular Weight Antimicrobial Compounds 394

5.3.6 Other Antimicrobial Agents 394

5.4 Immune Effects of Probiotic Bacteria 395

5.4.1 The Neonatal Intestinal Microbiota 395

5.4.2 The Importance of the Intestinal Microbiota in Immune Development 395

5.4.3 Interaction of Commensal and Pathogenic Bacteria with the Intestinal Immune System 396

5.4.4 Probiotic Effects on Immune Responses 396

5.4.5 Probiotic Effects on Epithelial Cells 397

5.4.6 Probiotic Effects on DCs 397

5.4.7 Probiotic Effects on Adaptive Immune Responses: T Helper Cells and T Regulatory Cells 397

5.4.8 Delivery of Probiotic Bacteria 398

5.4.9 The Specificity of Probiotic Effects 399

5.4.10 Summary 399

5.5 Alteration of Microecology in Human Intestine 399

5.5.1 Impact on Human Health: in Infants and the Elderly 399

5.5.1.1 Stepwise Establishment of Microbiota 400

5.5.1.2 Methodological Improvements in Microbiota Assessment 401

5.5.1.3 Microbiota After Infancy 403

5.5.1.4 Host–Microbe Cross Talk 403

5.5.1.5 Microbiota in the Elderly 404

5.5.1.6 Maintenance of Healthy Microbiota 405

5.5.1.7 Conclusion 405

5.5.2 Impact on Animal Health: Designer Probiotics for the Management of Intestinal Health and Colibacillosis in Weaner Pigs 406

5.5.2.1 The Farrowing Environment 406

5.5.2.2 The Weaning Environment 406

5.5.2.3 Colibacillosis in Pigs 407

5.5.2.4 Control of Colibacillosis 408

5.5.2.5 Mechanism of Action 408

5.5.2.6 Pathogenic and Commensal E. coli—the Concept of Gene Signatures 409

5.5.2.7 Mosaicism and Genome Plasticity in Porcine E. coli (Clone Gene Signatures) 410

5.5.2.8 Population Gene Signatures in Epidemiological Study 412

5.5.2.9 Designer Lactic Acid Bacteria as Probiotics 415

5.5.2.10 Population Gene Signatures as a Measure of Probiotic Bioefficacy 417

5.5.2.11 Creation of Enteric Microbial Communities for Sustainable Intestinal Health (Probiosis) 419

References 421

6 Commercially Available Human Probiotic Microorganisms 441

6.1 Lactobacillus acidophilus, LA-5^® 441

6.1.1 Gastrointestinal Effects 441

6.1.1.1 Intestinal Microbial Balance 441

6.1.1.2 Diarrhea 442

6.1.1.3 Other Gastrointestinal Effects 442

6.1.2 Immunomodulatory Effects 443

6.1.2.1 Nonspecific Immune Responses 443

6.1.2.2 Specific Immune Responses 443

6.1.3 Other Health Effects 443

6.1.4 Safety 444

6.2 Lactobacillus acidophilus NCDO 1748 444

6.2.1 Origin and Safety 445

6.2.2 In Vitro and Animal Studies 445

6.2.3 Human Studies 446

6.3 Lactobacillus acidophilus NCFM^® 447

6.3.1 L. acidophilus NFCM Basic Properties 447

6.3.2 Survival of Intestinal Transit and Change in Intestinal Microbiota Composition and Activity 447

6.3.3 Lactose Intolerance 448

6.3.4 Relief of Intestinal Pain 448

6.3.5 Prevention of Common Respiratory Infections and Effects on Immunity 449

6.3.6 Application 449

6.3.7 Conclusion 449

6.4 Lactobacillus casei Shirota 449

6.4.1 Effects on Intestinal Environment 450

6.4.2 Adhesive Property 450

6.4.3 Intestinal Physiology 451

6.4.4 Immunomodulation 452

6.4.5 Effects on Cancer 453

6.4.6 Prevention of Infectious Diseases 454

6.4.7 Prevention of Life Style Diseases 454

6.4.8 Clinical Application 455

6.4.9 Safety Assessment 456

6.5 Lactobacillus gasseri OLL2716 (LG21) 457

6.5.1 Helicobacter pylori 458

6.5.2 Selection of a Probiotic for H. pylori Infection 458

6.5.3 Effects of LG21 on H. pylori Infection in Humans 458

6.5.4 Mechanisms of Therapeutic Effects of LG21 on H. pylori Infection 461

6.5.5 Conclusion 462

6.6 Lactobacillus paracasei ssp. paracasei, F19^® 462

6.6.1 Identification and Safety 462

6.6.2 In Vitro Studies 463

6.6.3 Global Gene Expression 463

6.6.4 Human Studies 464

6.7 Lactobacillus paracasei ssp paracasei, L. casei 431^® 466

6.7.1 Adhesion and Survival Through the GI Tract 466

6.7.2 Gastrointestinal Effects 466

6.7.2.1 Intestinal Microbial Balance 466

6.7.2.2 Diarrhea 466

6.7.3 Immunomodulatory Effects 468

6.7.4 Other Health Effects 468

6.7.5 Safety 468

6.8 Lactobacillus rhamnosus GG, LGG^® 469

6.8.1 Storage Stability 469

6.8.2 Gastrointestinal Persistence and Colonization 469

6.8.3 Health Benefits 469

6.8.4 Source of LGG^® 470

6.9 Lactobacillus rhamnosus, GR-1^® and Lactobacillus reuteri RC- 14^® 470

6.9.1 The Strains 471

6.9.2 In Vitro Properties 471

6.9.3 Animal Safety, Toxicity, and Effectiveness Studies 471

6.9.4 Clinical Evidence 472

6.9.4.1 Safety, Effectiveness, and Efficacy 472

6.9.5 Summary 473

6.10 Lactobacillus rhamnosus HN001 and Bifidobacterium lactis HN019 473

6.10.1 Basic Properties of L. rhamnosus HN001 and B. lactis HN019 473

6.10.2 Survival During the Intestinal Transit and Modulation of the Intestinal Microbiota 474

6.10.3 Modulation of the Immune System 474

6.10.4 Reduction of Disease Risk 477

6.10.5 Application 477

6.10.6 Conclusions 477

6.11 LGG^®Extra, A Multispecies Probiotic Combination 477

6.11.1 Strain Selection for the Combination 477

6.11.2 Adhesion and Gastrointestinal Survival 478

6.11.3 Health Benefits 478

6.11.4 Technological Characteristics 479

6.11.5 Source of LGG^® Extra 480

6.12 Bifidobacterium animalis ssp. lactis, BB-12^® 480

6.12.1 Adhesion and Survival Through the GI Tract 480

6.12.2 Gastrointestinal Effects 480

6.12.2.1 Intestinal Microbial Balance 480

6.12.2.2 Diarrheas 481

6.12.2.3 Gastrointestinal Health of Infants 482

6.12.2.4 Other Gastrointestinal Effects 482

6.12.3 Immunomodulatory Effects 483

6.12.3.1 Nonspecific Immune Responses 483

6.12.3.2 Specific Immune Responses 483

6.12.3.3 Other Immunomodulatory Effects 484

6.12.4 Other Health Effects 484

6.12.5 Safety 485

6.13 Bifidobacterium breve Strain Yakult 485

6.13.1 Effects on Intestinal Environment 485

6.13.2 Intestinal Physiology 485

6.13.3 Effects on Cancer 486

6.13.4 Prevention of Infectious Diseases 486

6.13.5 Prevention of Life Style Diseases 486

6.13.6 Clinical Application 487

6.14 Bifidobacterium longum BB536 488

6.14.1 Evaluation of Safety of BB536 488

6.14.2 Physiological Effects of BB536 489

6.14.2.1 Improvement of Intestinal Environment 489

6.14.2.2 Effects on Immunity and Cancer 490

6.14.2.3 Antiallergic Activity 490

6.14.3 Technologies in BB536 Applications 491

6.15 Bifidobacterium longum Strains BL46 and BL2C—Probiotics for Adults and Ageing Consumers 492

6.15.1 Safety of BL2C and BL46 492

6.15.2 The Health Effects of BL2C and BL46 493

6.15.2.1 BL2C and BL46 Stabilize the Gut Function in the Elderly 493

6.15.2.2 Modulation of Gut Microbiota by BL2C and BL46 493

6.15.2.3 BL46 is Effective Against Harmful Bacteria 493

6.15.2.4 Effects of BL2C and BL46 on the Immune System and Infections 493

6.15.2.5 BL2C and BL46 Can Bind Toxic Compounds 493

6.15.3 Technical Properties and Sensory Qualities of BL2C and BL46 494

6.15.4 Conclusions 494

References 494

Part II Prebiotics 533

7 Prebiotics 535

7.1 The Prebiotic Concept 535

7.2 A Brief History of Prebiotics 536

7.3 Advantages and Disadvantages of the Prebiotic Strategy 536

7.4 Types of Prebiotics 537

7.5 Production of Prebiotics 540

7.6 Prebiotic Mechanisms 546

7.7 Modulating the Intestinal Microbiota in Infants 546

7.7.1 Breast Milk 546

7.7.2 Infant Milk Formulas 547

7.8 Modulating the Intestinal Microbiota in Adults 548

7.8.1 Effects at the Genus Level 548

7.8.2 Effects at the Species Level 548

7.8.3 Altering the Physiology of the Microbiota 549

7.9 Modifying the Intestinal Microbiota in the Elderly 549

7.10 Health Effects and Applications of Prebiotics 549

7.10.1 Laxatives 550

7.10.2 Hepatic Encephalopathy 550

7.10.3 Primary Prevention of Allergy in Infants 551

7.10.4 Amelioration of Inflammatory Bowel Disease 551

7.10.5 Prevention of Infections 555

7.10.6 Mineral Absorption 556

7.10.7 Prevention of Colorectal Cancer 556

7.10.8 Reduction in Serum Lipid Concentrations 559

7.10.9 Use in Weight Management and Improving Insulin Sensitivity 559

7.11 Functional Foods for Animals 559

7.12 Safety of Prebiotics 560

7.13 Regulation of Prebiotics 560

7.14 Conclusion 561

References 562

Author index 583

Subject index 585