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Pharmaceutical-Microbiology.Pdf This page intentionally left blank Copyright © 2008 New Age International (P) Ltd., Publishers Published by New Age International (P) Ltd., Publishers All rights reserved. No part of this ebook may be reproduced in any form, by photostat, microfilm, xerography, or any other means, or incorporated into any information retrieval system, electronic or mechanical, without the written permission of the publisher. All inquiries should be emailed to [email protected] ISBN (13) : 978-81-224-2867-4 PUBLISHING FOR ONE WORLD NEW AGE INTERNATIONAL (P) LIMITED, PUBLISHERS 4835/24, Ansari Road, Daryaganj, New Delhi - 110002 Visit us at www.newagepublishers.com In the recent past, the world has witnessed the smooth transition of the ‘quantum state’ of a trapped Ca2+ ion to another Ca2+ ion via meticulously teleported means in a critically controlled manner thereby mustering enough hope that in the near future ‘teleportation’ of individual atoms and molecules would pave the way to teleportation of molecules and ‘microorganisms’. ‘‘Dreams will only help you actualize your goals. I started dreaming when I was a child..... I became a scientist because I dreamt’’. —APJ Abdul Kalam Hon’ble President of India This page intentionally left blank 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) Microbiological (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. (vii) (viii) Microbial assays for antibiotics, vitamins, and amino acids have been treated at length with sufficient experimental details to enable students, research scholars, and budding scientists to pursue their objectives in the field of Pharmaceutical Microbiology. The author earnestly believes that ‘Pharmaceutical Microbiology’ may prove to be of paramount importance for B. Pharm. (Pharmacy Degree), M. Sc., (Food Microbiology), M. Sc., (Microbiology), and M. Sc., (Environmental Science) students as well. I extend my sincere thanks to Shri Saumya Gupta, MD and his excellent production wing to have the project completed in a record time frame. Gurgaon Ashutosh Kar 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) (x) 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 (xi) 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 (xii) 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] ..
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