The Project Gutenberg eBook of The Elements of Bacteriological Technique, by John William Henry Eyre This eBook is for the use of anyone anywhere at no cost and with almost no restrictions whatsoever. You may copy it, give it away or re-use it under the terms of the Project Gutenberg License included with this eBook or online at www.gutenberg.org Title: The Elements of Bacteriological Technique A Laboratory Guide for Medical, Dental, and Technical Students. Second Edition Rewritten and Enlarged. Author: John William Henry Eyre Release Date: January 5, 2009 [eBook #27713] Language: English Character set encoding: ISO-8859-1 ***START OF THE PROJECT GUTENBERG EBOOK THE ELEMENTS OF BACTERIOLOGICAL TECHNIQUE*** E-text prepared by Suzanne Lybarger, Brian Janes, Josephine Paolucci, and the Project Gutenberg Online Distributed Proofreading Team (http://www.pgdp.net) Transcriber's note: For numbers and equations: parentheses have been added to clarify fractions. Minor typographical errors have been corrected. THE ELEMENTS OF BACTERIOLOGICAL TECHNIQUEA LABORATORY GUIDE FOR MEDICAL, DENTAL, AND TECHNICAL STUDENTS BYJ. W. H. EYRE, M.D., M.S., F.R.S. (Edin.) Director of the Bacteriological Department of Guy's Hospital, London, and Lecturer on Bacteriology in the Medical and Dental Schools; formerly Lecturer on Bacteriology at Charing Cross Hospital Medical School, and Bacteriologist to Charing Cross Hospital; sometime Hunterian Professor, Royal College of Surgeons, England SECOND EDITION REWRITTEN AND ENLARGED PHILADELPHIA AND LONDON W. B. SAUNDERS COMPANY 1913 Copyright, 1902, by W. B. Saunders and Company Revised, entirely reset, reprinted, and recopyrighted July, 1913 Copyright, 1913, by W. B. Saunders Company Registered at Stationers' Hall, London, England PRINTED IN AMERICA PRESS OF W. B. SAUNDERS COMPANY PHILADELPHIA TO THE MEMORY OF JOHN WICHENFORD WASHBOURN, C.M.G., M.D., F.R.C.P. Physician to Guy's Hospital and Lecturer on Bacteriology in the Medical School, and Physician to the London Fever Hospital MY TEACHER, FRIEND, AND CO-WORKERPREFACE TO THE SECOND EDITION Bacteriology is essentially a practical study, and even the elements of its technique can only be taught by personal instruction in the laboratory. This is a self-evident proposition that needs no emphasis, yet I venture to believe that the former collection of tried and proved methods has already been of some utility, not only to the student in the absence of his teacher, but also to isolated workers in laboratories far removed from centres of instruction, reminding them of forgotten details in methods already acquired. If this assumption is based on fact no further apology is needed for the present revised edition in which the changes are chiefly in the nature of additions—rendered necessary by the introduction of new methods during recent years. I take this opportunity of expressing my deep sense of obligation to my confrère in the Physiological Department of our medical school—Mr. J. H. Ryffel, B. C., B. Sc.—who has revised those pages dealing with the analysis of the metabolic products of bacterial life; to successive colleagues in the Bacteriological Department of Guy's Hospital, for their ready co-operation in working out or in testing new methods; and finally to my Chief Laboratory Assistant, Mr. J. C. Turner whose assistance and experience have been of the utmost value to me in the preparation of this volume. I have also to thank Mrs. Constant Ponder for many of the new line drawings and for redrawing a number of the original cuts. John W. H. Eyre. Guy's Hospital, S. E. July, 1913.PREFACE TO THE FIRST EDITION In the following pages I have endeavoured to arrange briefly and concisely the various methods at present in use for the study of bacteria, and the elucidation of such points in their life-histories as are debatable or still undetermined. Of these methods, some are new, others are not; but all are reliable, only such having been included as are capable of giving satisfactory results even in the hands of beginners. In fact, the bulk of the matter is simply an elaboration of the typewritten notes distributed to some of my laboratory classes in practical and applied bacteriology; consequently an attempt has been made to present the elements of bacteriological technique in their logical sequence. I make no apology for the space devoted to illustrations, nearly all of which have been prepared especially for this volume; for a picture, if good, possesses a higher educational value and conveys a more accurate impression than a page of print; and even sketches of apparatus serve a distinct purpose in suggesting to the student those alterations and modifications which may be rendered necessary or advisable by the character of his laboratory equipment. The excellent and appropriate terminology introduced by Chester in his recent work on "Determinative Bacteriology" I have adopted in its entirety, for I consider it only needs to be used to convince one of its extreme utility, whilst its inclusion in an elementary manual is calculated to induce in the student habits of accurate observation and concise description. With the exception of Section XVII—"Outlines for the Study of Pathogenic Bacteria"—introduced with the idea of completing the volume from the point of view of the medical and dental student, the work has been arranged to allow of its use as a laboratory guide by the technical student generally, whether of brewing, dairying, or agriculture. So alive am I to its many inperfections that it appears almost superfluous to state that the book is in no sense intended as a rival to the many and excellent manuals of bacteriology at present in use, but aims only at supplementing the usually scanty details of technique, and at instructing the student how to fit up and adapt apparatus for his daily work, and how to carry out thoroughly and systematically the various bacterioscopical analyses that are daily demanded of the bacteriologist by the hygienist. Finally, it is with much pleasure that I acknowledge the valuable assistance received from my late assistant, Mr. J. B. Gall, A. I. C., in the preparation of the section dealing with the chemical products of bacterial life, and which has been based upon the work of Lehmann. John W. H. Eyre. Guy's Hospital, S. E.CONTENTS Page I. Laboratory Regulations 1 II. Glass Apparatus in Common Use 3 The Selection, Preparation, and Care of Glassware, 8—Cleaning of Glass Apparatus, 18—Plugging Test-tubes and Flasks, 24. III. Methods of Sterilisation 26 Sterilising Agents, 26—Methods of Application, 27—Electric Signal Timing Clock, 38. IV. The Microscope 49 Essentials, 49—Accessories, 57—Methods of Micrometry, 61. V. Microscopical Examination of Bacteria and Other Micro-fungi 69 Apparatus and Reagents used in Ordinary Microscopical Examination, 69—Methods of Examination, 74. VI. Staining Methods 90 Bacteria Stains, 90—Contrast Stains, 93—Tissue Stains, 95—Blood Stains, 97—Methods of Demonstrating Structure of Bacteria, 99—Differential Methods of Staining, 108. VII. Methods of Demonstrating Bacteria in Tissues 114 Freezing Method, 115—Paraffin Method, 117—Special Staining Methods for Sections, 121. VIII. Classification of Fungi 126 Morphology of the Hyphomycetes, 126—Morphology of the Blastomycetes, 129. IX. Schizomycetes 131 Anatomy, 134—Physiology, 136—Biochemistry, 144. X. Nutrient Media 146 Meat Extract, 148—Standardisation ofMedia, 154—The Filtration of Media, 156—Storing Media in Bulk, 159—Tubing Nutrient Media, 160. XI. Ordinary or Stock Culture Media 163 XII. Special Media 182 XIII. Incubators 216 XIV. Methods of Cultivation 221 Aerobic, 222—Anaerobic, 236. XV. Methods of Isolation 248 XVI. Methods of Identification and Study 259 Scheme of Study, 259—Macroscopical Examination of Cultivations, 261—Microscopical Methods, 272—Biochemical Methods, 276—Physical Methods, 295—Inoculation Methods, 315—Immunisation, 321—Active Immunisation, 322—The Preparation of Hæmolytic Serum, 327—The Titration of Hæmolytic Serum, 328—Storage of Hæmolysin, 331. XVII. Experimental Inoculation of Animals 332 Selection and Care of Animals, 335 —Methods of Inoculation, 352. XVIII. The Study of Experimental Infections During Life 370 General Observations, 371—Blood Examinations, 373—Serological Investigations, 378—Agglutinin, 381—Opsonin, 387—Immune Body, 393. XIX. Post-mortem Examination of Experimental Animals 396 XX. The Study of the Pathogenic Bacteria 408 XXI. Bacteriological Analyses 415 Bacteriological Examination of Water, 416—Examination of Milk, 441—Ice Cream, 457—Examination of Cream and Butter, 457—Examination of Unsound Meats, 460—Examination of Oysters and Other Shellfish, 463—Examination of Sewage and Sewage Effluents, 466—Examination of Air, 468—Examination of Soil, 470—Testing Filters, 478—Testing of Disinfectants, 480.Appendix 492 Index 505
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