Table Of Content

New Features Make the Micro Lab More Engaging and Applicable to Real Life Two newly added features in the Tenth Edition of Microbiology: A Laboratory Manual speak directly to student interest and engagement: A new experiment on hand washing and a new Clinical Applications feature that appears in each experiment. Hand-Washing Experiment A brand-new first experiment emphasizes the importance of effective hand washing and helps introduce microbiology in an engaging way. By learning about the presence of microorganisms on their own hands, this experiment gives students a practical understanding of why aseptic techniques in the lab are so important. A fun, practical lab, this experiment also gets students excited about the course to come. Clinical Applications Students often wonder how learning microbiological lab techniques will help them in their future careers. New Clinical Applications added to each experiment make the connection between lab, lecture, and future careers in nursing or other allied health fields. CLINICAL APPLICATION C GTccsocwtrshtatuueLbriuhhhaeinaslstlletealltIa p tum Gub ssapNiaeprnre rtieepmca irdae nS,saIdptt mdageiaa,Ciotre n y sd on amntncs ah,pAdibie ttlnunmaiea.r e no cion qLiia pisnrnrhncdutipel e ee ag iembeqAmsn sls:c o cu ota loTtfahadPiiirr mrrohdboteyoeeinPi oie emd.aifcr .nlnnC a gueLO fF .apodlnia idnoiTbInireorrdscf hCseistis cl,deeaecy ttdtaeAri n e ictrfmeDslnhtrsel isyTtostesbiiseiu natmefi iuraImboieigscmnl Oiatt sn pabginswicctinotu i anoNoowt ihernoerigfpstneedhen ra tstipin ,ie hnnai icor iecdegtlitohf n s c e g rgTec2t Gtef,ecaohdhwree ex°telvtrsaaesdnoce CapnhRscadttehiTumprtdw Coh onne.er Cttemrubnii hlaioacineL c os dt ooft ratfetsbtra ia his ,Lerlreiynssahot licl t eloiptt geid fap teetd rcann mtItieiaamr4emeeerhes s-eatgaNoor.tei Ror l-2a,rntu pirldmafiaese°e ettse Itdst’Cmodpssies rCoo pf it, agspiss c inuaasfntr aAteooa ubotantgtfrhloe rlawendd faiLe Gor Tdnlmn t uteft-ctg guoh tosr-tpsaeo Arel cr leieieaneKn t klictsarae.t riehP ias co ratHllnp eectotl liniPcbudlooe dlhwlsclnear.pwr hLeaegi Les cuntioest iress tlmIsttoarve asombtChtewtrae -i eiaininptrogyartAe,ln oie ohn hs asrbgdvrw lmlTo peeoayr emcmo av wt lI4cdsua owet me.Ootu trnn4rgoen e neye° dvorrrbs-NC c toeaacn3teh.eo wrgnys6a oyr dt egarothdn oeshe edfosoegla rao bbgu Mfde Isooadvt wnipwtaeerlf nthunufaelowaoso tycluneaacaslhionr,wko,yxvites ullthntwiietidb eiltb h,rnsbeacsgu t r,aiaaiiegri pg itsmcalaen ss-l edelya-.lfo iel tfop cdoooshe srrvwDfafecooe as,a ttsrbsosneymum i p etnaoioganuppaelfn g l ioTstsrtec heigtiryphaeihte linoluclsreeerit fntnha her ipds cia pentsdayro ypstterutnri uiiycdffygseefog e.esnrpo r,gCg c tafmwd.i rot tskasrahhmmau.dyo ingRcbe gaba-hie enrglem ssenrar eoeusyatPa iidcsmaosrseuitlnnc-eladcon ,hutn ewehwt dthosge ohb a uaortmiasaasctvi ckepoehvset iehnr eoet a oaharfscl iwsedl ae n- lnl Photos of Protozoa, Yeast, and Fungi Parts 6 and 7 of the manual now contain even more photos and micrographs of protozoa, yeast, and fungi to help students visualize these important organisms in the lab. ® MasteringMicrobiology Takes the Microbiology Laboratory Course to the Next Level The Mastering online homework, assessment, and tutorial system delivers self-paced tutorials that provide students with individualized coaching set to the instructor’s course objectives. MasteringMicrobiology® helps students arrive better prepared for lecture and lab with practice quizzes, coaching activities, tutorials, and more. Instructors can now assign MicroLab Tutors, MicroLab Practicals, Lab Technique Videos, and Lab-specific Quizzes within the MasteringMicrobiology item library. Students can access the MicroLab Tutors, MicroLab Practicals and Lab Technique Videos for review and self-study in the StudyArea of MasteringMicrobiology. MicroLab Tutors help instructors and students get the most out of lab time by providing addi- tional opportunities to reinforce labconcepts and techniques. MicroLab Tutors introduce important lab techniques and include each technique’s background, purpose, and clinical applications before walking students through the procedure itself. Using a combination of micrographs, video footage in the lab, and, for select topics, 3D molecular animations, MicroLab Tutors help students visualize important lab techniques and processes and make the connection between lecture and lab. MicroLab Practical assessments give students extra practice in observing and analyzing test results–putting knowledge into practice. These Lab Practical questions give users an extra opportunity to analyze and interpret important lab tests, procedures, and results in order to get ready for the lab practical! Lab Technique Videos are 3–5 minute videos, demonstrating specific lab techniques. These videos cover important procedures, such as aseptic technique, Gram staining, and smear preparation. The videos and their associated assessments help students prepare for the wet lab and allow them to review techniques on their own time. Lab-Specific Quizzes are newly created for each experiment in the manual. A total of ten multiple-choice and true/false questions per experiment allow instructors to quiz students on microbiological principles, lab techniques, and the theory behind lab procedures. Each quiz question is tied to a Learning Objective for the lab. What Instructors are Saying “These are the types of lab questions I am looking for. Hallelujah!” ▶ Pele Rich North Hennepin Community College “I think they are really great!! It puts things in a place where the students can observe firsthand what they should be seeing. I definitely would use them.” ▶ Tanya Crider East Mississippi Community College Pearson Custom Library: Lab Manuals Tailored for Your Course WithThe Pearson Custom Library for the Biological Sciences, instructors can create a lab manual that meets their specific content needs and organization exactly. But that’s not all! Instructors now have the power to customize their favorite Pearson microbiology lab manuals. View and select just the labs you need, in the sequence you want, deleting experiments you don't use: Your students pay only for the materials you choose. You can also include your own original content, like your syllabus, course sched- ule or unique labs. Then preview your custom manual online (with net price and ISBN), and request an evalua- tion copy to examine before you order. The Pearson Custom Library is the perfect option to put you in control of your course materials. To create the perfect lab manual, visit: www.pearsoncustomlibrary.com Build Your Manual in Three Easy Steps In just three easy steps, you can build your ideal customized lab manual that is aligned to your teaching objectives and designed to promote student achievement. SEARCH (cid:116)(cid:1) Search Pearson’s vast library of content, including Pearson textbooks, lab manuals, editorially-selected third party readings, and cases. (cid:116)(cid:1) Search by author, title, topic, or content type. SELECT & PERSONALIZE (cid:116)(cid:1) Select only the experiments you use and arrange them in the order you want. (cid:116)(cid:1) Upload any original content. (cid:116)(cid:1) Choose from a set of diverse and engaging covers. PREVIEW & FINALIZE (cid:116)(cid:1) Preview Table of Contents (cid:116)(cid:1) View a digital review copy instantaneously, or have a bound print copy sent to you. (cid:116)(cid:1) Once you decide to adopt, use your automatically generated custom ISBN to place an order at your campus store. 540 Contents M I C R O B I O L O G Y A L A B O R A T O R Y M A N U A L TENTH EDITION James G. Cappuccino SUNY Rockland Community College Natalie Sherman SUNY Rockland Community College Boston Columbus Indianapolis New York San Francisco Upper Saddle River Amsterdam Cape Town Dubai London Madrid Milan Munich Paris Montréal Toronto Delhi Mexico City São Paulo Sydney Hong Kong Seoul Singapore Taipei Tokyo Acquisitions Editor: Kelsey Churchman Director of Development: Barbara Yien Senior Project Development Editor: Marie Beaugureau Editorial Assistant: Ashley Williams Managing Editor: Deborah Cogan Production Project Manager: Megan Power Production Management and Composition: Integra Interior Designer: Jerilyn Bockorick Cover Designer: Yvo Riezebos Image Management: Travis Amos Manufacturing Buyer: Stacey Weinberger Executive Marketing Manager: Neena Bali Text Printer: Webcrafters, Inc. Cover Printer: Lehigh-Phoenix Color Cover Photo Credit: Andreas Reh / Getty Images Credits and acknowledgments borrowed from other sources and reproduced, with permission, in this textbook appear on the appropriate page within the text or on p. 531. Copyright © 2014, 2011, 2008 Pearson Education, Inc. All rights reserved. Manufactured in the United States of America. This publication is protected by Copyright, and permission should be obtained from the publisher prior to any prohibited reproduction, storage in a retrieval system, or transmission in any form or by any means, electronic, mechanical, photocopying, recording, or likewise. To obtain permission(s) to use material from this work, please submit a written request to Pearson Education, Inc., Permissions Department, 1900 E. Lake Ave., Glenview, IL 60025. For information regarding permissions, call (847) 486-2635. Many of the designations used by manufacturers and sellers to distinguish their products are claimed as trademarks. Where those designations appear in this book, and the publisher was aware of a trademark claim, the designations have been printed in initial caps or all caps. Library of Congress Cappuccino, James G. Microbiology : a laboratory manual/James G. Cappuccino, Natalie Sherman.—10th ed. p.; cm. Includes bibliographical references and index. ISBN-13: 978-0-321-84022-6 ISBN-10: 0-321-84022-4 I. Sherman, Natalie. II. Title. [DNLM: 1. Microbiology—Laboratory Manuals. QW 25] LC Classification not assigned 579.078—dc23 2012037762 Student edition ISBN 10: 0-321-84022-4 ISBN 13: 978-0-321-84022-6 Instructor’s Review Copy ISBN 10: 0-321-88451-5 ISBN 13: 978-0-321-88451-0 www.pearsonhighered.com 1 2 3 4 5 6 7 8 9 10 11 12—WBC—17 16 15 14 13 Contents Preface ix PART 3 Bacterial Staining 55 Laboratory Safety xiii Introduction 55 Laboratory Protocol xv Experiment 8: Preparation of Bacterial Smears 59 PART 1 Basic Laboratory Techniques Experiment 9: Simple for Isolation, Cultivation, and Staining 65 Cultural Characterization of Experiment 10: Negative Microorganisms 1 Staining 71 Experiment 11: Gram Stain 75 Introduction 1 Experiment 12: Acid-Fast Experiment 1: Effectiveness of Stain 81 Hand Washing 7 Experiment 13: Differential Experiment 2: Culture Transfer Staining for Visualization of Techniques 13 Bacterial Cell Structures 87 Experiment 3: Techniques for Part A: Spore Stain (Schaeffer- Isolation of Pure Cultures 19 Fulton Method) 87 Part A: Isolation of Discrete Part B: Capsule Stain (Anthony Colonies from a Mixed Method) 89 Culture 19 Part B: Isolation of Pure PART 4 Cultivation of Microorganisms: Cultures from a Spread-Plate or Streak-Plate Preparation 22 Nutritional and Physical Experiment 4: Cultural Requirements, and Characteristics of Enumeration of Microbial Microorganisms 29 Populations 97 Introduction 97 PART 2 Microscopy 35 Experiment 14: Nutritional Introduction 35 Requirements: Media for the Experiment 5: Microscopic Routine Cultivation of Examination of Stained Cell Bacteria 99 Preparations 37 Experiment 15: Use of Experiment 6: Microscopic Differential, Selective, and Examination of Living Enriched Media 105 Microorganisms Using a Experiment 16: Physical Factors: Hanging-Drop Preparation or Temperature 115 a Wet Mount 45 Experiment 17: Physical Experiment 7: The Factors: pH of the Extracellular Microscopic Measurement of Environment 121 Microorganisms 51 iii Experiment 18: Physical PART 6 The Protozoa 221 Factors: Atmospheric Oxygen Introduction 221 Requirements 125 Experiment 34: Free-Living Experiment 19: Techniques for Protozoa 223 the Cultivation of Anaerobic Microorganisms 131 Experiment 35: Parasitic Protozoa 229 Experiment 20: Serial Dilution– Agar Plate Procedure to Quantitate Viable Cells 137 PART 7 The Fungi 237 Experiment 21: The Bacterial Introduction 237 Growth Curve 145 Experiment 36: Cultivation and Morphology of Molds 239 PART 5 Biochemical Activities of Part A: Slide Culture Microorganisms 153 Technique 239 Introduction 153 Part B: Mold Cultivation on Solid Surfaces 242 Experiment 22: Extracellular Enzymatic Activities of Experiment 37: Yeast Morphology, Microorganisms 155 Cultural Characteristics, and Reproduction 247 Experiment 23: Carbohydrate Fermentation 161 Experiment 38: Identification of Unknown Fungi 255 Experiment 24: Triple Sugar–Iron Agar Test 167 Experiment 25: IMViC Test 171 PART 8 The Viruses 261 Part A: Indole Production Introduction 261 Test 172 Experiment 39: Cultivation Part B: Methyl Red Test 173 and Enumeration of Part C: Voges-Proskauer Bacteriophages 265 Test 174 Experiment 40: Isolation of Part D: Citrate Utilization Coliphages from Raw Sewage 271 Test 175 Experiment 26: Hydrogen Sulfide PART 9 Physical and Chemical Agents Test 183 for the Control of Microbial Experiment 27: Urease Test 187 Growth 277 Experiment 28: Litmus Milk Reactions 191 Introduction 277 Experiment 29: Nitrate Reduction Experiment 41: Physical Agents Test 197 of Control: Moist Heat 279 Experiment 30: Catalase Experiment 42: Physical Agents Test 201 of Control: Environmental Osmotic Experiment 31: Oxidase Pressure 285 Test 205 Experiment 43: Physical Agents Experiment 32: Utilization of of Control: Electromagnetic Amino Acids 209 Radiations 289 Part A: Decarboxylase Test 209 Experiment 44: Chemical Agents of Control: Chemotherapeutic Part B: Phenylalanine Agents 293 Deaminase Test 211 Part A: The Kirby-Bauer Experiment 33: Genus Antibiotic Sensitivity Test Identification of Unknown Procedure 294 Bacterial Cultures 215 iv Contents Part B: Synergistic Effect of Experiment 55: Bacterial Drug Combinations 296 Conjugation 371 Experiment 45: Determination of Experiment 56: Isolation Penicillin Activity in the Presence of a Streptomycin-Resistant and Absence of Penicillinase 303 Mutant 377 Experiment 46: Chemical Agents Experiment 57: The Ames Test: A of Control: Disinfectants and Bacterial Test System for Chemical Antiseptics 309 Carcinogenicity 381 PART 10 Microbiology of Food 317 PART 14 Biotechnology 387 Introduction 317 Introduction 387 Experiment 47: Microbiological Experiment 58: Bacterial Analysis of Food Products: Transformation 389 Bacterial Count 319 Experiment 59: Isolation of Experiment 48: Wine Bacterial Plasmids 397 Production 323 Experiment 60: Restriction Analysis and Electrophoretic PART 11 Microbiology of Water 327 Separation of Bacteriophage Lambda DNA 407 Introduction 327 Experiment 49: Standard PART 15 Medical Microbiology 417 Qualitative Analysis of Water 329 Experiment 50: Quantitative Introduction 417 Analysis of Water: Membrane Filter Experiment 61: Microbial Flora Method 337 of the Mouth: Determination of Susceptibility to Dental Caries 419 PART 12 Microbiology of Soil 343 Experiment 62: Normal Microbial Flora of the Throat and Skin 423 Introduction 343 Experiment 63: Identification Experiment 51: Microbial of Human Staphylococcal Populations in Soil: Pathogens 431 Enumeration 345 Experiment 64: Identification Experiment 52: Isolation of Human Streptococcal of Antibiotic-Producing Pathogens 439 Microorganisms and Experiment 65: Identification of Determination of Antimicrobial Streptococcus pneumoniae 447 Spectrum of Isolates 351 Experiment 66: Identification Part A: Isolation of Antibiotic- of Enteric Microorganisms Using Producing Microorganisms 352 Computer-Assisted Multitest Part B: Determination of Microsystems 453 Antimicrobial Spectrum of Experiment 67: Isolation and Isolates 353 Presumptive Identification of Experiment 53: Isolation of Campylobacter 463 Pseudomonas Species by Means Experiment 68: Microbiological of the Enrichment Culture Analysis of Urine Specimens 467 Technique 357 Experiment 69: Microbiological Analysis of Blood Specimens 473 PART 13 Bacterial Genetics 363 Experiment 70: Species Identification of Unknown Introduction 363 Bacterial Cultures 479 Experiment 54: Enzyme Induction 365 Contents v