كتاب Pharmaceutical microbiology

PREFACE The textbook of ‘Pharmaceutical Microbiology’ specifically aims at the ever demanding thoughtful need of an absolutely well-documented compilation of factual details related to : theoritical principles, classifications, diagramatic profiles, graphic presentations, critical explanation, latest examples for the Pharmacy Degree (B. Pharm.,) throughout the Indian Universities, SAARC-countries, and similar curricula adopted abroad. Modern invigorative society, based on the overwhelming and overemphasized broad-spectrum importance vis-a-vis utilities of ‘Microbiology’ profusely gets benefited from the intricate species of scores of microorganisms in several ways and means, namely : antibiotics, vaccines, enzymes, vitamins etc. Nevertheless, a quantum-leap-forward in the field of ‘Modern Biotechnology’ rests predominantly upon reasonably sound microbiological foundation. Besides, microorganisms do modulate a plethora of vital and critical functionalities, such as : ( a ) enable completion of cycles of C, O, N and S which essentially occur in both terrestrial and aquatic systems ; ( b ) provide absolutely indispensable components of prevailing ecosystem ; and ( c ) serve as a critical source of ‘nutrients’ occurring at the grass-root of practically a large segment of ecological food webs and chains. The entire course-content presented in ‘Pharmaceutical Microbiology’ has been meticulously and painstakingly developed and expanded as per the AICTE-Approved Syllabus–2000. Each chapter has been duly expatiated in a simple, lucid, and crisp language easily comprehensible by its august readers. A unique largely acceptable style of presentation has been adopted, viz., brief introduction, principles, labeled figures, graphics, diagrams of equipments, descriptions, explanations, pharmaceutical applications, and selected classical examples. Each chapter is duly elaborated with adequate foot-notes, references, and ‘further reading references’ at the end. An exhaustive ‘Glossary of Important Microbiological Terminologies’ has been duly annexed at the end of the textbook. A fairly up to date computer-generated ‘Index’ in the textbook will surely enlarge the vision of its readers in gaining an easy access of subject enriched well documented text materials. Pharmaceutical Microbiology consists of Ten Chapters : (1) Introduction and Scope ; (2) Structure and Function : Bacterial Cells ; (3) Characterization, Classification and Taxonomy of Microbes ; (4) Identification of Microorganisms ; (5) Nutrition, Cultivation and Isolation : Bacteria- Actinomycetes-Fungi-Viruses ; (6) Microbial Genetics and Variations ; (7) Microbial Control by Physical and Chemical Methods ; (8) Sterility Testing : Pharmaceutical Products ; (9) Immune Systems ; and (10) Mi crobiological (Microbial) Assays : Antibiotics–Vitamins–Amino Acids. The text material essentially embodies not only an ample emphasis on the vivid coverage of fundamental principles of microbiology as a scientific discipline but also maintains a manageable length for the apprehension of brilliant students. CONTENTS 1. Introduction and Scope ... 1 1.1 Introduction ... 1 1.2 Historical Development of Microbiology ... 3 1.2.1. The Microscope ... 3 1.2.2. Spontaneous Generation Vs Biogenesis ... 4 1.2.3. Fermentation ... 6 1.2.4. Germ Theory ... 6 1.2.5. Classical Laboratory Methods and Pure Cultures ... 7 1.2.6. Immunity ... 8 1.2.7. Medical Microbiology ... 9 1.2.8. Pharmaceutical Microbiology ... 10 1.2.9. Industrial Microbiology ... 14 1.2.10. Emergence of Molecular Biology ... 15 1.2.11. Emergence of Virology ... 17 1.2.12. Microorganisms as Geochemical Agents ... 19 1.2.13. Microbiology in the New Millennium ... 19. 2. Structure and Function : Bacterial Cells ... 23 2.1 Introduction ... 23 2.2 Characteristic Features ... 23 2.2.1. Shape ... 23 2.2.2. Size ... 23 2.2.3. Reproduction ... 24 2.2.4. Formation of Colony ... 24 2.2.5. Mutation ... 24 2.2.6. Motility ... 24 2.2.7. Food and Oxygen Requirements ... 24 2.2.8. Temperature Requirements ... 24 2.3 Activities ... 25 2.4 Organization of Microbial Cells ... 25 2.4.1. Type of Cells ... 26 2.4.1.1. Eukaryotic Cells ... 27 2.4.1.2. Prokaryotic Cells ... 33 2.5 Archaeobacteria and Eubacteria ... 37 2.5.1. Methanogenic Bacteria [Methanogens] ... 38 2.5.2. Extreme Halophiles ... 40 ( ix ) 2.5.3. Thermoacidophiles ... 41 2.5.3.1. Thermoplasma ... 41 2.5.3.2. Sulfolobus ... 41 2.6 The Bacterial Cells ... 42 2.6.1. Typical Bacterial Cells ... 43 2.6.2. Capsules and Slimes ... 44 2.6.3. Flagella and Fimbria ... 46 2.6.3.1. Flagella ... 46 2.6.3.2. Fimbria [or Pili] ... 48 2.6.4. Cell Envelope ... 49 2.6.5. Gram-Positive and Gram-Negative Bacteria ... 51 2.6.6. Significance of Teichoic Acids ... 53 2.6.7. The Cell Membrane ... 54 2.6.8. Bacterial Cytoplasm ... 55 2.6.9. Ribosomes ... 57 2.6.10. Cellular Reserve Materials ... 58 3. Characterization, Classification and Taxonomy of Microbes ... 62 3.1 Introduction ... 62 3.2 Characterization ... 62 3.2.1. Morphological Characteristics ... 63 3.2.2. Chemical Characteristics ... 64 3.2.3. Cultural Characteristics ... 64 3.2.4. Metabolic Characteristics ... 66 3.2.5. Antigenic Characteristics ... 66 3.2.6. Genetic Characteristics ... 67 3.2.6.1. DNA Base Composition ... 67 3.2.6.2. Sequence of Nucleotide Bases in DNA ... 68 3.2.7. Pathogenecity ... 69 3.2.8. Ecological Characteristics ... 69 3.3 Classificiation ... 70 3.3.1. Difficulties Encountered in Classification of Microorganisms ... 70 3.3.2. Objectives of Classification ... 70 3.3.3. Genetic Methods of Classifying Microbes ... 71 3.3.3.1. Genetic Relatedness ... 71 3.3.3.2. The Intuitive Method ... 72 3.3.3.3. Numerical Taxonomy ... 72 3.3.4. Systemetized Classification ... 75 3.3.4.1. Natural Classification ... 75 3.3.4.2. Phyletic Calssification ... 75 ( x ) 3.3.4.3. Linnear Binomial Scheme ... 76 3.3.4.4. Phenotypic Classification ... 77 3.3.4.5. Microscopic Examination ... 79 3.3.4.6. Cataloguing rRNA ... 80 3.3.4.7. Computer Aided Classification ... 81 3.3.4.8. Bacterial Classification ... 82 3.4 Taxonomy ... 87 3.5 The Kingdom Prokaryotae ... 88 3.5.1. Actinomyctes ... 89 3.5.1.1. General Characteristics ... 89 3.5.1.2. Significance of Actinomycetes ... 90 3.5.1.3. Classification ... 91 3.5.1.3.1. Whole Cell Carbohydrate Patterns of Aerobic Actinomycetes ... 91 3.5.1.3.2. Major Constituents of Cell Wall Types of Actinomycetes ... 91 3.5.1.3.3. Groups of Actinomycetes Based on Whole Cell Carbohydrate Pattern and Cell Wall Type ... 92 3.5.1.3.4. Actinomycetes with Multiocular Sporangia ... 92 3.5.1.4. Actinomycetes and Related Organisms ... 93 3.5.1.4.1. Group ... 93 3.5.1.4.2. Genus ... 94 3.5.1.4.3. Order ... 97 3.5.1.4.4. Family ... 98 3.5.2. Bacteria ... 102 3.5.2.1. Salient Features ... 103 3.5.2.2. Structure and Form of the Bacterial Cell ... 104 3.5.2.2.1. Size and Shape ... 105 3.5.2.2.2. Structure ... 105 3.5.3. Rickettsia and Coxiella ... 107 3.5.4. Spirochaetes ... 108 4. Identification of Microorganisms ... 112 4.1 Introduction ... 112 4.2 Morphology ... 113 4.3 Selective and Diagnostic Media ... 113 4.3.1. Differential Media ... 116 4.3.1.1. Eosin Methylene Blue Agar [EMB-Agar] ... 116 ( xi ) 4.3.1.2. MacConkey Agar ... 116 4.3.1.3. Hektoen Enteric Agar [HE-Agar] ... 116 4.3.2. Enrichment Media ... 116 4.3.2.1. Blood Agar ... 116 4.3.2.2. Chocolate Agar ... 117 4.3.3. Characteristic Media ... 117 4.3.3.1. Triple Sugar Iron Agar [TSI-Agar] ... 117 4.4 Cultural Characteristics ... 119 4.5 Biochemical Tests (or Properties) ... 120 4.5.1. Carbohydrate (Sugar) Fermentation ... 120 4.5.2. Litmus Milk ... 120 4.5.3. Indole Production ... 120 4.5.4. Methyl Red Test [MR-Test] ... 121 4.5.5. Voges-Proskauer Test [VP-Test] ... 121 4.5.6. Citrate Utilization ... 121 4.5.7. Nitrate Reduction ... 122 4.5.8. Ammonia Production ... 122 4.5.9. Urease Test ... 122 4.5.10. Production of Hydrogen Sulphide ... 123 4.5.11. Reduction of Methylene Blue ... 123 4.5.12. Production of Catalase [Tube Catalase Test] ... 123 4.5.13. Oxidase Reaction ... 123 4.5.14. Egg-Yolk Reaction ... 124 4.5.15. Growth in Presence of Potassium Cyanide ... 124 4.5.16. Composite Media ... 124 4.6 Profile of Microbial Stains ... 127 4.6.1. Preparation of Bacterial Specimens for Light Microscopy ... 128 4.6.1.1. Standard Preparations ... 128 4.6.1.2. Preparation of Smears for Staining ... 128 4.6.1.3. Gram Staining ... 129 4.6.1.4. Differential Staining ... 131 4.6.1.4.1. Gram’s Stain ... 131 4.6.1.4.2. Acid-Fast Stain ... 131 4.6.1.5. Miscellaneous Staining ... 131 4.6.1.5.1. Capsule Staining ... 132 4.6.1.5.2. Endospore Staining ... 132 4.6.1.5.3. Flagella Staining ... 133 ( xii ) 4.6.2. Microscopy : The Differential Instruments ... 133 4.6.2.1. Concepts ... 133 4.6.2.2. Microscope Variants ... 134 4.6.2.2.1. Bright-Field Microscope ... 134 4.6.2.2.2. Dark-Field Microscope ... 136 4.6.2.2.3. Phase-Contrast Microscope ... 136 4.6.2.2.4. Differential Interference Contrast (DIC) Microscope ... 139 4.6.2.2.5. Fluorescence Microscope ... 139 4.6.2.2.6. Electron Microscope ... 141 4.6.2.2.6.1. Transmission Electron Microscope (TEM) ... 142 4.6.2.2.6.2. Scanning Electron Microscope (SEM) ... 143 5. Nutrition, Cultivation and Isolation : Bacteria-Actinomycetes-Fungi-Viruses ... 146 5.1 Introduction ... 146 5.2 Bacteria ... 146 5.2.1. Nutrition of Microorganisms ... 146 5.2.2. Cultivation of Bacteria ... 147 5.2.2.1. Binary Fission ... 148 5.2.2.2. Normal Growth Curve of Microorganisms ... 149 5.2.2.3. The Lag Phase of Microbial Growth ... 150 5.2.2.4. Translational Periods Between Various Growth Phases ... 150 5.2.2.5. Synchronous Growth ... 151 5.2.2.6. Effect of Nutritional Concentration Vs Growth Rate of Bacterial Culture ... 152 5.2.2.7. Growth Determining Techniques ... 152 5.2.3. Isolation of Bacteria ... 154 5.2.3.1. Selective and Diagnostic Media ... 154 5.2.3.2. Bismuth Sulphate Agar ... 154 5.2.3.3. Selective Media for Staphylococci ... 155 5.3 Actinomycetes ... 155 5.4 Fungi ... 156 5.4.1. Reproduction of Fungi ... 158 5.4.1.1. Asexual Reproduction ... 158 5.4.1.2. Sexual Reproduction ... 159 5.4.2. Industrial Importance of Fung Methods and specifications Testing of pharmaceutical products is carried out according to a Pharmacopeia of which there are a few types. For example: In America, the United States Pharmacopeia is used; in Japan there is the Japanese Pharmacopeia; in the United Kingdom there is the British Pharmacopoeia and in Europe the European Pharmacopeia. These contain a test method which is to be followed when testing, along with defined specifications for the amount of microorganisms allowed in a given amount of product. The specifications change depending on the product type and method in which it is introduced to the body. The pharmacopoeia also covers areas like sterility testing, endotoxin testing, the use of biological indicators, microbial limits testing and enumeration, and the testing of pharmaceutical grade water.
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