Table Of ContentNew 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 microbiol-
ogy 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 prac-
tice 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 impor-
tant lab techniques and include
each technique’s background,
purpose, and clinical applications
before walking students through the procedure itself. Using a combination of mi-
crographs, 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 ques-
tions 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 as-
sessments 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 microbiologi-
cal 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
