Wine Microbiology: Practical Applications and Procedures / Edition 2

by Kenneth C. Fugelsang

Overview

Since publication of the first edition of Wine Microbiology in 1997, the volume of new information and concepts has dramatically increased. Even with the tremendous increase in available information, a comprehensive understanding regarding the role of individual microorganisms towards wine quality as well as the impact of complicated interactions between microorganisms and processing techniques is lacking.

Wine Microbiology, Second Edition, fills this void. Like the first edition, the book addresses real world problems such as characterization and enumeration of yeast, bacteria and molds common to juice and wine environments and their impact on wine quality and stability. In addition, the potential of solving processing problems through rapid, real-time molecular methods and new applications for using starter cultures of non-Saccharomyces yeast are discussed. The book is a great resource for professionals and students in the field of enology and viticulture.

About the Author:
Kenneth C. Fugelsang has been associated with California State University (Fresno) since 1970 and is now winemaster and professor of enology

About the Author:
Charles G. Edwards has been a professor with the Department of Food Science and Human Nutrition at Washington State University

Product Details

ISBN-13:	2900387333419
Publication date:	11/14/2006
Pages:	394
Product dimensions:	6.50(w) x 1.50(h) x 9.50(d)

About the Author

Kenneth C. Fugelsang has been associated with California State University (Fresno) since 1970 and is now winemaster and professor of enology. He is an author of 12 books and over 100 technical papers and presentations. Professor Fugelsang has been recipient of the Claude Laval Award for Innovative Technology, the University Award for Research and Scholarly Activity, as well as the Outstanding Alumnus Award from the College of Science and Mathematics. In addition to teaching and research responsibilities, he is responsible for operation of the university’s commercial 50,000 gallon winery where student–produced wines have received over 150 super, gold, and silver awards at national and international wine competitions.

Charles G. Edwards received his Bachelor of Science, Master of Science, and Doctor of Philosophy degrees in food science from Oregon State University (1982), Cornell University (1985), and The Pennsylvania State University (1989), respectively. He is currently a professor with the Department of Food Science and Human Nutrition at Washington State University (Pullman, Washington, USA). Dr. Edwards is an author of 150+ technical papers and presentations and is sole author of Illustrated Guide to Microbes and Sediments in Wine, Beer, and Juice. His laboratory was award the best research paper published in the American Journal of Enology and Viticulture in 1991 and best student presentations at the American Society for Enology and Viticulture annual meeting in 2004. Dr. Edwards currently teaches courses in food chemistry and wine microbiology/processing.

Table of Contents

Preface to the Second Edition     xv
Acknowledgments     xvii
Introduction     xix
Grape and Wine Microorganisms
Yeasts     3
Introduction     3
Reproduction     3
Sexual Reproduction     4
Asexual Reproduction     4
Taxonomy     7
Candida     8
Dekkera     9
Hanseniaspora     11
Issatchenkia     11
Metschnikowia     11
Pichia     12
Saccharomyces     12
Saccharomycodes     13
Schizosaccharomyces     14
Zygosaccharomyces     14
Nutritional Requirements     15
Nitrogen     16
Growth and Survival Factors     17
Metabolism     18
Glucose     18
Sulfur     23
Odor/Flavor Compounds     23
Glycosidases     26
Mannoproteins     28
Lactic Acid Bacteria     29
Introduction     29
Taxonomy     29
Lactobacillus     30
Oenococcus     32
Pediococcus     33
Nutritional Requirements     34
Metabolism     36
Glucose     36
Arginine     38
Malate     39
Mannitol and Erythritol     40
Diacetyl and Other Odor/Flavor Compounds     41
Acetic Acid Bacteria     45
Introduction     45
Taxonomy     45
Nutritional Requirements     47
Metabolism     47
Carbohydrates     47
Ethanol     49
Molds and Other Microorganisms     52
Introduction     52
Ecology Habitats     53
Taxonomy     54
Aspergillus (Black Mold)     54
Botrytis (Gray Mold)     54
Penicillium (Blue-Green Mold)     56
4.4 Nutritional Requirements     56
4.5 Metabolism     56
Glucose     56
Mycotoxins     57
Odor/Flavor Compounds     58
Other Microorganisms     59
Bacillus     59
Clostridium     59
Streptomyces     60
Vinification and Winery Processing
Managing Microbial Growth     65
Introduction      65
Preservatives and Sterilants     65
Sulfur Dioxide     66
Dimethyl Dicarbonate     70
Lysozyme     72
Sorbic Acid     73
Other Preservatives and Sterilants     75
Filtration     77
Perpendicular-Flow Filtration ("Nominal" or "Depth")     77
Perpendicular-Flow Filtration ("Absolute" or "Sterile")     78
Cross-Flow/Tangential Filtration     80
Microbial Ecology During Vinification     82
Introduction     82
Non-Saccharomyces and Saccharomyces Yeasts     84
Grapes and Musts     84
Alcoholic Fermentation     85
Post-fermentation     87
Dekkera/Brettanomyces     87
Grapes and Musts     87
Alcoholic and Post-fermentation     87
Lactic Acid Bacteria     88
Grapes and Musts     88
Alcoholic and Malolactic Fermentations     89
Post-fermentation     91
Acetic Acid Bacteria     91
Grapes and Musts     91
Alcoholic and Post-fermentation     91
Microbial Interactions     93
Saccharomyces and Oenococcus     93
Saccharomyces and Lactobacillus     96
Oenococcus, Pediococcus, and Lactobacillus     98
Other Interactions     100
Harvest and Pre-fermentation Processing     102
Introduction     102
Harvest and Transport     102
Fruit Quality Assessment     103
Soluble Solids     103
pH and Titratable Acidity     104
Microbial Spoilage     105
Must Processing     106
Enzymes     106
Suspended Solids     107
Pre-fermentation Maceration (Cold-Soak)     108
Thermovinification     109
Inert Gassing     109
Processing Microbially Deteriorated Fruit     110
Enzymatic Browning     110
Fermentation Difficulties     111
Clarification Concerns     112
Management Strategies     112
Juice Storage (Mute)     113
Fermentation and Post-fermentation Processing     115
Introduction     115
Must Supplementation     115
Alcoholic Fermentation     118
Historical Perspective of Starter Cultures     118
Preparation of Starter Cultures     119
Strain Selection      121
Temperature     122
Immobilized Yeast     122
"Natural" Fermentations     123
Fermentation Problems     124
Sluggish/Stuck Fermentations     124
Hydrogen Sulfide     128
Malolactic Fermentation     130
Preparation of Starter Cultures     130
Strain Selection     132
Timing of Inoculation     132
Use of Schizosaccharomyces     135
Post-Fermentation Processing     135
Aging and Storage     135
Adjustment of Volatile Acidity     136
Bottling     137
Winery Cleaning and Sanitizing     139
Introduction     139
Safety Issues     140
Water Quality     140
Preliminary Cleaning     141
Detergents, Cleaners, and Surfactants     142
Alkali     142
Acids     143
Surfactants     143
Rinses     143
Sanitizers     144
Iodine     145
Quaternary Ammonium Compounds     145
Acidulated Sulfur Dioxide     146
Peroxides     147
Chlorine and Chlorinated Formulations      148
Hot Water and Steam     148
Ozone     149
Sanitation Monitoring     150
Schedules and Documentation     151
Quality Points Program     153
Introduction     153
Developing QP Programs     154
Processing and Flowchart     155
Quality Factor Analysis     156
Examples of Quality Factors     156
Preventive Measures     157
Critical Quality Points     157
Critical Limits     158
Monitoring Procedures     159
Corrective Actions     160
Record-Keeping and Documentation     160
Verification     161
Wine Spoilage     162
Introduction     162
Spoilage Microorganisms     162
Acetohacter     162
Dekkera/Brettanomyces     164
Film Yeasts     166
Saccharomycodes     167
Zygosaccharoviyces     168
Wine Faults     168
Volatile Acidity     168
Ethyl Carbamate     170
Mousiness     172
Post-MLF Bacterial Growth     172
Geranium Odor/Tone      173
Biogenic Amines     174
Acrolein     176
Mannitol     178
Ropiness     178
Tartaric Add Utilization     179
Laboratory Procedures and Protocols
Basic Microscopy     183
Introduction     183
Microscopes     183
Magnification     184
Resolution     184
Contrast     185
Fluorescence Microscopy     185
Using Microscopes     187
Components     187
General Use     189
Calibration     189
Preparing Smears     190
From Liquid Media     190
From Solid Media     191
Preparing Wet Mounts     191
Preparing Mold Slide Cultures     192
Media Preparation and Culture Techniques     194
Introduction     194
Physical/Chemical Requirements for Cultivation     195
Carbon and Nitrogen     195
Oxygen     196
Hydrogen Ion Concentration (pH)     196
Moisture and Water Activity     197
Incubation Temperature and Conditions     198
Selective Agents     198
Sterilization of Laboratory Media and Supplies     199
Boiling Water     199
Steam Sterilization     200
Dry Heat     201
Sterile Filtration     202
Chemical Sterilization     202
Storage of Prepared Media     204
Media for Yeasts and Molds     205
Wort Medium     206
Grape Juice Medium     207
WL Medium     207
Brettanomyces Medium A     207
Brettanomyces Medium B     208
Brettanomyces Medium C     208
Brettanomyces Medium D     209
Lysine Medium     209
Zygosaccharomyces Medium     210
Media for Lactic Acid Bacteria     211
Apple Juice Rogosa Medium     211
Tomato Juice-Glucose-Fructose-Malate Medium     212
Heterofermentation-Arginine Broth     212
Media for Acetic Acid Bacteria     213
Glucose-Yeast Extract-Carbonate Medium     213
Mannitol-Yeast Extract-Peptone Medium     214
Yeast Extract-Peptone-Ethanol Medium     214
Aseptic Transfer Techniques     214
Transfers from Solid to Solid Media     215
Transfers from Solid to Liquid Media     216
Transfers from Liquid to Solid Media     217
Isolation of Microorganisms     219
Maintenance and Storage of Cultures     221
Preparation of Agar Slants and Stabs     221
Glycerol Suspensions     221
Freeze-drying (Yeasts)     221
Freeze-drying (Lactic Acid Bacteria)     222
Estimation of Population Density     224
Introduction     224
Sampling     225
Sample Dilution     226
Direct Microscopic Count     228
Using a Microscope Counting Chamber     229
Methylene Blue     231
Ponceau-S     232
Wolford's Stain     232
Direct Plating     233
Pour Plates     234
Spread Plates     235
Membrane Filtration     236
Bioluminescence     238
Nephelometry and Optical Density     239
Identification of Wine Microorganisms     241
Introduction     241
Identifying Microorganisms     242
Yeasts and Molds     244
Assimilation of Carbon and Nitrogen     244
Demonstration of Ascospores     250
Demonstration of Mycelia/Pseudomycelia      252
Fermentation of Carbohydrates     253
Bacteria     254
Ammonia from Arginine     255
Catalase     256
Dextran from Sucrose     257
Fermentation of Carbohydrates     257
Gas from Glucose     260
Gram Stain     262
Ketogenesis     264
Lactate from Glucose     264
Malate Utilization (Monitoring MLF)     266
Mannitol from Fructose     269
Oxidation of Ethanol     269
Oxidation of Lactate     271
Other Technologies for Identification and Enumeration     273
Introduction     273
Phenotypical Identification     274
Biolog System     274
Fatty Acid Methyl Ester Analysis     274
Protein Characterization     275
Electrophoretic Characterization of Isozymes (Zymograms)     275
Immunochemical Techniques     276
Enzyme-Linked Immunosorbent Assay     276
Immunochemical Fluorescence Microscopy     278
Phylogenetic Analyses     278
Polymerase Chain Reaction     279
Gel Electrophoresis     281
Quantitative Polymerase Chain Reaction      282
Hybridization Probes     283
Fluoresence In Situ Hybridization     283
Ribonucleic Acid Analysis     283
Probe Detection Systems     284
TaqMan Probes     284
Molecular Beacons     284
Scorpions     286
Peptide Nucleic Acid Chemiluminescent In Situ Hybridization Probes     287
Other Molecular Methods     287
Restriction Fragment Length Polymorphisms     287
Pulsed Field Gel Electrophoresis     288
Additional Methods     288
Extrachromosomal Elements (Satellites)     289
Chemical and Physical Instabilities     290
Introduction     290
Crystalline Instabilities     290
Tartrates     290
"Crystal-Like" Instabilities     293
Cork Dust     293
Diatomaceous Earth     294
Fibrous Instabilities     295
Amorphous Instabilities     296
Protein     296
Phenolics     298
Glucans, Pectins, and Starch     299
Metal Casse     300
Laboratory Setup     303
Introduction     303
Microscope and pH Meter      304
Autoclave     305
Centrifuge and Filters     305
Incubators     306
Water Baths     306
Glass and Plasticware     307
Media     308
Photometers     308
Laminar Flow Hoods     308
Miscellaneous     309
Laboratory Safety     311
Introduction     311
Injury and Illness Prevention Program     311
Training     312
Information     312
Communication     314
Eyewash Stations and Safety Showers     314
Fire Alarms and Prevention     316
First Aid     316
Personal Protective Equipment     316
Examples of Safety Issues     318
Biohazard and Chemical Waste     319
Electricity     319
Heat and Steam     319
Machinery Safeguards     320
Storage     320
Ultraviolet Light     320
Glossary     322
References     331
Index     381

Editorial Reviews

Fugelsang (enology, California State U.-Fresno) provides a step-by- step procedural guide to the isolation and characterization of bacteria and yeast common to the juice and wine environment, discussing not only routine methodology, but also the interpretation of results and how they affect the winemaker. He outlines specific factors, including sanitation, that impact the winemaking process at all stages from the primary processing of grapes through the must, juice, and fermentation, to bottling. He also describes the equipment and supplies needed to add a microbiological unit to an existing analytical laboratory. Annotation c. by Book News, Inc., Portland, Or.

Booknews

From the reviews of the second edition:

"This second edition takes into account the tremendous increase in the amount of information that has become available since the publication of the first edition. … Given that the authors are authorities in their field, this book is self recommending." (Wine East, 2007)

From the Publisher