Michael Rieth received his biology doctorate from the Institute of Microbiology University of Gottingen. He entered into the pharmaceutical industry in 1988. From 1999 to 2014 he was Head of Biological Quality Assurance at Merck KGaA, Darmstadt. He then worked in the field of Global Regulatory Affairs until his retirement in 2020. He has written four books, including Pharmaceutical Microbiology in the German language, which was awarded the Wallhausser Prize in 2013.

Preface to the English Edition xvii

Preface to the First German Edition xix

List of Abbreviations xxi

Glossary xxvii

1 Introduction to Microbiology 1

1.1 Historical 1

1.2 Importance 2

1.3 World of Microorganisms 3

1.4 The Bacterial Cell 9

1.5 Taxonomy of Microorganisms 18

1.6 Medical Microbiology 22

References 26

Further Reading 27

2 General Conditions for the Operation of Microbiological Laboratories 29

2.1 Laws and Technical Regulations 29

2.2 Medical Care for Employees 33

2.3 Operating Description for Microbiological Laboratories 36

2.4 Establishment of Microbiological Laboratories 37

2.4.1 Required Devices/Equipment 37

2.5 Nutrient Media 39

2.6 Media Preparation 47

References 78

3 Calibration and Qualification of the Devices 81

3.1 Calibration Error 83

3.2 Calibration 83

3.3 Balance 84

3.4 pH Meter 85

3.5 Piston Pipettes 85

3.6 Stopwatch 86

3.7 Devices for Achieving Specific Temperatures 87

3.8 Clean Bench 90

3.9 Air Sampler 93

3.10 Particle Counter 95

3.11 Measuring Device for Determining the Water Activity 96

3.12 Photometer/Reader 96

3.13 Tube Reader for Endotoxin Determinations 98

3.14 Fluorescence Reader for Endotoxin Determinations 98

References 98

4 Stock Collection 101

4.1 Reference 101

4.2 Shipping 105

4.3 Storage 105

4.4 Cultivation 106

References 107

5 Industrial Hygiene 109

5.1 Hygiene 109

5.2 Microbiological Basics for Hygiene 112

5.3 Hygiene Measures 115

5.4 Sterilization, Disinfection, and Aseptic Production 127

5.5 Hygiene Protocol for Microbiological Laboratories 136

5.6 Pest Control 137

5.7 Hygiene Officer 140

5.8 Implementation of Hygiene Training Courses 140

References 143

6 Environmental Monitoring 145

6.1 Methods 145

6.2 Microbiological Monitoring in the Sterility Test Isolator 150

6.3 Physical Monitoring in Sterile Production 154

6.4 Physical Operation 155

6.5 Evaluation of the Microorganisms 158

6.6 Register of Microorganisms 159

References 170

Further Reading 172

7 Quality Control 173

7.1 Pharmacopeial Methods (Compendial Methods) 173

7.2 Non-Compendial Methods 214

7.3 Tests Using Animal Models 228

7.4 Cell Culture Methods 231

7.5 Validation of Pharmacopeial Methods 235

References 263

Further Reading 266

8 Process Validations 267

8.1 Media Fill 267

8.2 Depyrogenation 274

8.3 Validation of Sterilization with Dry Heat 276

8.4 Validation of Sterilization Using Moist Heat (Autoclave) 278

8.5 Validation of Sterile Filtration 279

8.6 Container Closure Integrity Test 282

8.7 Cleaning Validation 286

References 289

Further Reading 290

9 Microbiological Examination of Water 291

9.1 Sampling 293

9.2 Specimen Transport 294

9.3 Use of the Different Water Qualities (Table 9.6) 297

9.4 Purified Water 297

9.5 Highly Purified Water 301

9.6 Water for Injection 301

9.7 Water for Diluting Concentrated Hemodialysis Solutions 302

9.8 Water for the Preparation of Extracts 302

9.9 Drinking Water 303

9.10 Legionella 305

References 308

10 Rapid Microbiological Methods 309

10.1 Determination via ATP Content 309

10.2 Determination Via the Incorporation of Fluorescent Markers 313

10.3 Flow Cytometry 315

References 316

Further Reading 317

11 Automation in the Microbiology Laboratory 319

11.1 Automatic Dyeing Machines 319

11.2 Devices for Counting Colonies 320

11.3 Automatic Nutrient Media Filling Machine 320

11.4 Automation of the Endotoxin Test 321

References 322

12 Quality Assurance 323

12.1 Structure of an SOP System 323

12.2 Training 324

12.3 Audits and Inspections 326

12.4 Procedure for OOS and OOE Results 330

References 337

13 Microorganism Identification 339

13.1 Growth Curve 339

13.2 Generation Time 340

13.3 Preparation of Pure Cultures 340

13.4 Sensory and Macroscopic Characteristics 340

13.5 Microscopic Examination 341

13.6 Staining 347

13.7 Principle of the "Colored Row" 349

13.8 Immunological Procedures 351

13.9 Polymerase Chain Reaction 351

13.10 Gas Chromatography (FAME) 352

13.11 FT-IR Spectroscopy 352

13.12 Maldi-tof 353

References 353

14 Cleaning, Sterilization, Decontamination, and Disposal 355

14.1 Cleaning 355

14.2 Sterilization 355

14.3 Laboratory Cleaning and Disinfection 360

14.4 Disposal of Infectious Waste 362

14.5 Disinfection Measures in Case of Accidents 363

References 363

15 Contract Testing (Outsourcing) 365

References 366

Further Reading 366

16 Microbiological Networks 367

16.1 CPM 367

16.2 VAAM Expert Group Quality Management 367

16.3 Subcommittee Microbiology in the VfA 368

16.4 DGHM 368

References 369

17 Addresses 371

Literature 379

Index 389