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Laboratory Applications in Microbiology A Case Study
Approach 2nd Edition Barry Chess Digital Instant
Download
Author(s): Barry Chess
ISBN(s): 9780073402376, 0073402370
Edition: 2
File Details: PDF, 88.39 MB
Year: 2011
Language: english
Laboratory Applications
in Microbiology
A CASE STUDY APPROACH

Second Edition

Barry Chess
Pasadena City College

TM
 TM

LABORATORY APPLICATIONS IN MICROBIOLOGY: A CASE STUDY APPROACH, SECOND EDITION

Avenue of the
Americas, New York, NY All rights r
Previous editions © 2009. No part of this publication may be reproduced or distributed in any form or by
any means, or stored in a database or r consent of The McGraw-Hill
electr
or broadcast for distance learning.

outside

ee paper.

1 2 3 4 5 6 7 8 9 0 QDB/QDB 1 0 9 8 7 6 5 4 3 2 1

ISBN 978-0-07-340237-6
MHID 0-07-340237-0

Vice President & Editor

Vice President EDP/Central Publishing Services: Kimberly Meriwether David
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Design Coor Brenda A. Rolwes
Cover Designer: Studio Montage, St. Louis, Missouri
Photo Research Coordinator: Lori Hancock
Cover Images: (Scientist Peering into Microscope): © Getty Images RF; (Arrangements of Rod-Shaped Bacteria,
Bacillus Anthracis Bacterial Colonies, Posteroanterior (PA) Chest X-Ray): © Center for Disease Control
Buyer: Susan K. Culbertson
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All credits appearing on page or at the end of the book are considered to be an extension of the

Some of the laboratory experiments included in this text may be hazardous if materials are handled improperly
or if procedures are conducted incorrectly. Safety precautions are necessary when you are working with
microorganisms, chemicals, glass test tubes, hot water baths, sharp instruments, and the like, or for any
procedures that generally require caution. Your school may have set regulations regar ocedures
oblems with materials or procedures, please
ask your instructor for help.
Contents
Preface vii MANIPULATION, STAINING, AND OBSERVATION
OF MICROORGANISMS
ESSENTIAL LABORATORY SKILLS
xerc ep c ec n ques
Case Study Exercise 1 Safety Considerations in the CASE SYNOPSIS
Microbiology Laboratory 1 Multiple Misdiagnoses of Tuberculosis Resulting from
CASE SYNOPSES Laboratory Error—Wisconsin, 1996
Laboratory-Acquired Infection with Escherichia coli O157: Case Study Exercise 8 Pure Culture Techniques 67
H7—New York, 2004
CASE SYNOPSIS
Ocular Vaccinia Infection of a Laboratory Worker—
Neonatal Tetanus—Montana, 1998
Philadelphia, 2004
Laboratory Researcher Dies after Suffering Burns—Los Case Study Exercise 9 Simple Staining, Negative
Angeles, California, 2009 Staining, and Gram Staining 79
Case Study Exercise 2 Microscopy and Measurement CASE SYNOPSIS
of Microscopic Specimens 9 Identification of Bacteria Responsible for the Outbreak of
Gastrointestinal Disease
CASE SYNOPSIS
Excerpt of Letters from Anton van Leeuwenhoek to the Case Study Exercise 10 Capsular Staining 91
Royal Society of London for the Improvement of Natural CASE SYNOPSIS
Knowledge Pneumococcal Sepsis after Autosplenectomy—2005
Case Study Exercise 11 Acid-Fast and Endospore
SURVEY OF MICROORGANISMS Staining 95
CASE SYNOPSES
Case Study Exercise 3 Identification and
Mycobacterium tuberculosis Transmission in a Newborn Nursery
Classification of Algae 21
and Maternity Ward—New York City, 2003
CASE SYNOPSIS Inhalation Anthrax Associated with Dried Animal Hides—
Oregon Harmful Algal Bloom Monitoring Project London, 2008
Case Study Exercise 4 Su ey of Medically Important
Protozoa 33
CASE SYNOPSIS ENVIRONMENTAL INFLUENCES ON THE GROWTH
Acanthamoeba keratitis—Multiple States
OF MICROORGANISMS

Case Study Exercise 5 Identification and xerc a e oun

Classification of Fungi 41 CASE SYNOPSIS
CASE SYNOPSIS Salmonella Typhimurium Infection Traced to Contaminated
Outbreak of Histoplasmosis among Travelers Returning from Milk
El Salvador, 2008 Case Study Exercise 13 Cultivation of Anaerobes 109
Case Study Exercise 6 Ubiquity of CASE SYNOPSIS
Microorganisms 53 Botulism Associated with Commercially Canned Chili Sauce—
CASE SYNOPSIS Texas and Indiana, July 2007
Sorcerer II Global Ocean Sampling Expedition—2007–2009 Case Study Exercise 14 Temperature Effects on
Bacterial Growth and Survival 117
CASE SYNOPSIS
Outbreak of Gastroenteritis Associated with Consumption of
Alaskan Oysters—2004

iii
iv Contents

Case Study Exercise 15 pH and Microbial Growth 123 Case Study Exercise 25 The Ames Test 203
CASE SYNOPSIS CASE SYNOPSIS
Botulism: Episode Leads to New FDA Regulation West Nile Virus Update—United States, January 1–December 31,
2009
Case Study Exercise 16 Effects of Osmotic Pressure
on Bacterial Growth 131
CASE SYNOPSIS
Vibrio vulnificus Infection Traced to Sewage Spill—Hawaii, 2006
APPLIED MICROBIOLOGY
Case Study Exercise 26 DNA Extraction from
Bacterial Cells 209
CONTROL OF MICROBIAL GROWTH CASE SYNOPSIS
At the Limits of Science: 9/11 ID Effort Comes to an End
Case Study Exercise 17 Lethal Effects of Ultraviolet
Case Study Exercise 27 DNA Profiling 213
Light 137
CASE SYNOPSIS
CASE SYNOPSIS
Multistate Outbreak of Salmonella Infections Associated with
Gastrointestinal Outbreak Traced to Interactive Fountain—
Peanut Butter and Peanut Butter-Containing Products—
New York, March 2006
United States, 2008–2009
Case Study Exercise 18 Evaluation of Disinfectants:
Case Study Exercise 28 Measures of Water Quality:
Use-Dilution Method 143
Most Probable Number Procedure 225
CASE SYNOPSIS
CASE SYNOPSIS
Anaphylaxis Following Cystoscopy Caused by a High-Level
E. coli Contamination of Water Supply—Frazier Park,
Disinfectant—2004
California, 2007
Case Study Exercise 19 Effectiveness of Hand
Case Study Exercise 29 Measures of Water Quality:
Scrubbing 151
Membrane Filtration Method 233
CASE SYNOPSIS
CASE SYNOPSIS
Puerperal Fever—Vienna, Austria, 1847
Fecal Contamination of Airline Drinking Water—2005
Case Study Exercise 20 Antimicrobic Sensitivity
Case Study Exercise 30 Measures of Milk Quality:
Testing: Kirby-Bauer, Tube Dilution, and E-Test
Methylene Blue Reductase Test 241
Methods 159
CASE SYNOPSIS
CASE SYNOPSES
Salmonella Typhimurium Infection Associated with Raw Milk
Methicillin-Resistant Staphylococcus aureus Infections among
and Cheeses Consumption—Pennsylvania, 2007
Tattoo Recipients—Ohio, Kentucky, and Vermont,
2004–2005 Case Study Exercise 31 Bacterial Counts of
New Antibiotic Discovered—Germany, 2008 Food 247
CASE SYNOPSIS
Food Poisoning among Inmates at a County Jail—Wisconsin,
EPIDEMIOLOGY August, 2008
Case Study Exercise 21 Phage Typing of Bacteria 171
CASE SYNOPSIS
Salmonella Serotype Enteritidis Infections among Workers MEDICAL MICROBIOLOGY
Producing Poultry Vaccine—Maine, November–December,
Case Study Exercise 32 Epidemiology of
2006
Gastrointestinal Illness: Differentiation of
Case Study Exercise 22 Simulated Epidemic 177 Enterobacteriaceae 253
CASE SYNOPSIS CASE SYNOPSIS
Import-Associated Measles Outbreak—Indiana, May–June, 2005 Gastroenteritis among Evacuees from Hurricane Katrina—
Houston, Texas, 2005
Case Study Exercise 23 Morbidity and Mortality
Weekly Report 185 Case Study Exercise 33 Isolation and Identification
CASE SYNOPSIS of Staphylococci 263
Google Used to Predict Influenza Outbreaks—2009 CASE SYNOPSIS
Methicillin-Resistant Staphylococcus aureus Skin Infections from
an Elephant Calf—San Diego, California, 2008
MICROBIAL GENETICS
Case Study Exercise 34 Isolation and Identification
Case Study Exercise 24 Bacterial of Streptococci 273
Transformation 193 CASE SYNOPSIS
CASE SYNOPSIS Invasive Streptococcus pyogenes after Allograft Implantation—
Multidrug-Resistant Acinetobacter Infections in Soldiers—2004 Colorado, 2003
 Contents v

Case Study Exercise 35 Blood Typing 285 Exercise 54 Mannitol Salt Agar 395
CASE SYNOPSIS Exercise 55 MacConkey Agar 399
Transfusion Reaction Leads to Death due to ABO
Exercise 56 Desoxycholate Agar 403
Incompatibility—Florida, 2008
Exercise 57 Endo Agar 407
Case Study Exercise 36 Differential White Blood
Cell Count 291 Exercise 58 Eosin Methylene Blue Agar 411
CASE SYNOPSIS Exercise 59 Hektoen Enteric Agar 415
Screening for Parasitic Infection of Refugees—United States, Exercise 60 Xylose Lysine Desoxycholate
2008 Agar 419
Case Study Exercise 37 Slide Agglutination 297 Exercise 61 Blood Agar 423
CASE SYNOPSIS Exercise 62 Motility Media 427
Leptospirosis Infection—Hawaii, 2005
Exercise 63 SIM Medium 431
Case Study Exercise 38 Enzyme-Linked
Exercise 64 Kligler’s Iron Agar 435
Immunosorbent Assay (ELISA) 301
CASE SYNOPSIS Exercise 65 Triple Sugar Iron Agar 439
Hepatitis C Virus Transmission at an Outpatient Hemodialysis Exercise 66 Lysine Iron Agar
Unit—New York, 2001–2008 Exercise 67 Litmus Milk 447

Commonly Used Biochemical Tests

IDENTIFICATION OF UNKNOWN BACTERIA
Exercise 68 Oxidation-Fermentation Test 451
Case Study Exercise 39 Identification of Bacterial Exercise 69 Phenol Red Broth 455
Unknowns 309
Exercise 70 Purple Broth 459
CASE SYNOPSIS
Respiratory Disease Strikes Legionnaires’ Convention— Exercise 71 Methyl Red and Voges-Proskauer
Philadelphia, 1976 Tests 463
Exercise 72 Catalase Test 467
Exercise 73 Oxidase Test 471
LABORATORY TECHNIQUES, REAGENTS, AND
ASSAYS Exercise 74 Nitrate Reduction Test 473
Exercise 75 Coagulase Test 477
Analysis of Bacterial Cultures Based on Morphological
Exercise 76 Citrate Test 479
Characteristics
Exercise 40 Colony Morphology 329 Exercise 77 Malonate Test 483

Exercise 41 333 Exercise 78 Decarboxylation Test 487

Exercise 79 Phenylalanine Deaminase Test 491
Staining Techniques Used for the Microscopic
Exercise 80 Bile Esculin Test 493
Examination of Bacteria
Exercise 81 Starch Hydrolysis 497
Exercise 42 Simple Stain 337
Exercise 82 ONPG Test 501
Exercise 43 Negative Stain 343
Exercise 83 Urease Test 505
Exer Capsule Stain 347
Exercise 84 Casease Test 509
Exercise 45 Gram Stain 351
Exercise 85 Gelatinase Test 513
Exercise 46 Endospore Stain 357
Exercise 86 DNase Test 517
Exercise 47 Acid-Fast Stain 361
Exercise 87 Lipase Test 519
Exercise 48 Motility Methods: Wet Mount and
Hanging Drop 365 Exercise 88 CAMP Test 523
Exercise 49 Flagella Stain 369 Exercise 89 PYR Test 525

Techniques for Inoculation of Media Commercial Identification Systems

Exercise 50 Streak-Plate Isolation 373 Exercise 90 API 20E System 527
Exercise 51 Loop Dilution 381 Exercise 91 Enterotube II System 535
Exercise 52 Spread-Plate 387
Antimicrobial Susceptibility Tests
Commonly Used Differential and Selective Media Exercise 92 Antibiotic Disk Sensitivity Tests 543
Exercise 53 Fluid Thioglycollate Medium 391 Exercise 93 𝛃-Lactamase Test 547
vi Contents

Quantitative Techniques Appendix C: Preparation of Culture Media 563

Exercise 94 Viable Plate Count 549 Appendix D: Media, Reagents, and Stain Formulas 568
Exercise 95 Direct Cell Count 555 Appendix E: Data Sheet for Unknown Identification 576
Glossary 577
Appendix A: Spectrophotometric Determination of Bacterial
Growth: Use of the Spectrophotometer 557 Credits 595
Appendix B: Use of Pipettes in the Laboratory 559 Index 597
Preface
“Did you see the interesting article in the newspaper Case Studies
about . . . ?” Every instructor of micr
lecture with these, or similar, words. And always for the same The first 39 exercises begin with actual cases taken from the
reason: to impress upon our students that microbiology is scientific literature. After reading each case, several ques-
not only important but is, in fact, relevant to all our lives. For tions help to define the most important issues and how they
microbiologists, and those who already intend to enter the should be addressed. As the exercise is completed, new
field, this battle of relevance has already been won, but for techniques, media, and observational tools are introduced,
obiology as all with the goal of solving the issues presented in the case.
a pr Evidence has shown that the use of case studies boosts
pr eases reten-
to everyday experiences. Usually we, as microbiologists, rec- tion of students in the classroom, and even reduces the inci-
dence of academic dishonesty
living example of the impact of micr more, learn faster, and retain more with case studies than
, news with traditional instruction methods. Although this seems
stories do not always adhere to our syllabi, and many of these e the day’s
news story with our class, the results are backed up by
uctors who
use cases, 97 percent reported that students who were
For Whom Is This Lab Manual taught with cases learned new ways to think about an issue;
Written? 95 percent reported that students took a more active part in
the learning process; and 92 percent reported that students
W Labora- were mor uly remarkable numbers.
Case Study Approach, is
designed to use real-life examples, or case studies, as the
basis for exercises in the laboratory. Over the past few years,
Changes to the Second Edition
the number of lecture texts utilizing case studies has grown The second edition of
rapidly, and for good reason—case studies work! This book has incorporated a number of new featur
is the only lab manual focusing on this means of instruction, keep the manual up-to-date, make it easier to use, and
an approach particularly applicable to the micr improve the experience of both students and instructors.
oratory. All the micr These changes include:
little if students cannot understand the importance of a
ocedure. • Five new exercises have been added to the manual,

What Sets This Lab Manual Apart? exer cises cover DNA profiling,
, differential white blood cell
eated to make the micr e count, slide agglutination, and the use of ELISA.
valuable experience by r what and how of • A e than 300 images has
microbiology with the sometimes forgotten why. Although
a clear visual reference for every exercise, test result,
always be a part of the curriculum, the context of the exer- and organism they will encounter.
cises has been expanded so that the r • Student learning outcomes (SLOs) have been included
specific task will be clear fr es of for the first 39 exercises in the manual, outlining the
the book are used to accomplish this goal and serve to dis- skills and theory a student should master as they
tinguish it from other microbiology lab manuals. complete each exercise.

vii
viii Preface

• Seventeen completely new case studies have been

revised to impr
information.
• The importance of laboratory safety has been -
emphasized through the inclusion of an additional case oce-
study to Exercise 1. dur
• Exer
has been moved so that it falls immediately after
exercises devoted to the identification of algae and e will r ocedures for
protozoa, providing a more unified look at eukaryotic A data sheet in the appendix provides
microorganisms. ecor esults, r
for ecor .
• The procedures in Exercise 17—Lethal Effects of
Ultraviolet Light—and Exercise 18—Evaluation of
Disinfectants—have been modified to produce more Extensive Flowcharts for Bacterial
consistent results. Identification
• Exer eekly
Exercise 39 introduces the concept of bacterial identification,
Reports—contains a particularly inter
Legionella
on the use of Google to track H1N1 influenza.
pneumophila es’ disease.
Additionally, Table 23.1 has been updated to reflect
Within this exercise, 31 flowcharts are used to help identify
bacterial unknowns commonly seen in the microbiology
• Recipes for media have been removed fr cise laboratory, a far more extensive collection than the one or
themselves but may still be found in Appendix D for two found in most manuals. This exercise also serves as an
those who desire them. intr
• All micr eference to the
ar e.
students to compare the images they obtain with those
in the book.
A Self-Contained Resource for the
Progression of Exercises Promotes Microbiology Laboratory
Active Learning
In the workplace, allied health professionals are expected to
evaluate a situation and find a solution using whatever
Material in each of the first 39 exercises has been car
resources are available to them. This book serves as a self-
or
contained resource, with everything a student needs to solve
ough the
a problem in the microbiology laboratory. A pro-
case study
vides definitions of all microbiological terms used in the
. Immediately following the
Appendices contain the formula
oductory material, pre-lab questions help stu-
of every media and reagent used, in addition to tutorials
dents to focus on the important aspects of the case, develop-
covering universal techniques such as the use of pipettes
ing a framework for what they will need to do prior to the lab,
and spectrophotometers as well as the preparation of media.
most of which require two or thr
Each exercise also includes a link to applicable websites,
multiday labs, questions are posed to ensure that the stu-
such as the CDC homepage for each pathogenic microor-
dents understand what they have just done, the results they
ganism encountered. In short, this book will help students
esults. Post-lab
develop the ability to solve problems.
questions requir om the

cise and the real-life application. Teaching and Learning Supplements

why
obiology, the how of the subject has not been for
ough compendium of
Digital Tools for Your Lab Course
cises are New! McGraw-Hill Connect Microbiology for Chess’
pr allows instruc-
tors and students to use art and animations for assignments
ferential and lectures. Instr
media or biochemical tests, are grouped together resources including assignable and gradable lab questions
exer . By r from the lab manual, additional pre- and post-lab activities,
 Preface ix

case study activities, interactive questions based on atlas your students are ready for an alternative, McGraw-Hill
eBooks offer a less expensive and eco-friendly alternative
uctor resources are available to traditional printed textbooks and laboratory manuals.
thr This laboratory manual is available as an eBook at www.
CourseSmart.com. At CourseSmart, your students can take
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downloaded to a computer. The eBooks allow students to

do full text searches, add highlighting and notes, and share
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resentative or visit www.CourseSmart.com to learn more.

Personalize Your Lab

Craft your teaching resour
With McGraw-Hill Create™, www.mcgrawhillcreate.com,
Blackboard, the web-based course management system, you can easily rearrange chapters, combine material from
ces, and quickly upload content you have
and faculty to use online materials and activities to comple- written like your course syllabus or teaching notes. Find
d features exciting the content you need in Create by searching through thou-
social learning and teaching tools that foster more logical, sands of leading McGraw-Hill textbooks. Arrange your
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ence how McGraw-Hill Create empowers you to teach your
Create, you also get deep integration of McGraw-Hill con- students your way.
tent and content engines right in Blackboard. Whether
you’r
assignments, all the tools you need are right where you Student Resource
want them—inside of Blackboard. Annual Editions: Micr 1 (0-07-738608-6) is a series
Gradebooks are now seamless. When a student com- of over 65 volumes, each designed to provide convenient,
inexpensive access to a wide range of current articles from
assignment automatically (and instantly) feeds your Black- some of the most respected magazines, newspapers, and
board grade center. journals published today. Annual Editions are updated on a
McGraw-Hill and Blackboard can now offer you easy regular basis through a continuous monitoring of over 300
periodical sources. The articles selected are authored by
your campus hosts it, or we do. Be sure to ask your local pr esearchers, and commentators writing
McGraw-Hill representative for details. Annual Editions volumes have a
Digital Lecture Capture. Tegrity Campus™ is a service
make them particularly useful in the classroom: a general
that allows class time to be any time by automatically cap-
introduction; an annotated table of contents; a topic guide;
turing every lecture in a searchable video format for stu-
orld Wide Web sites; and a
dents to review at their convenience. Educators know that
isit www
the more students can see, hear, and experience class
for more details.
resources, the better they learn. Help turn all your stu-
dents’ study time into learning moments by supplying
them with your lecture videos. Acknowledgments

Electronic Book—GO GREEN! (with apologies to all the moms out there), then I suppose
Green. . . . it’s on everybody’s mind these days. It’s not only producing the second edition is a bit like raising a child. You
about saving trees, it’s also about saving money. If you or want to be sur
x Preface

you send him on his way he r Debra M. Adair

not complaints. Well, so far there have been mostly compli- Paradise V
ments, the complaints have been generally minor, and to
take the par , it has, for sure, Anthony Arment
e due. Central State University

ar Michelle Badon
University of Texas at Arlington
e is no way I
ed feedback. Please Ranjit Banerjee
New York Medical College
PCC, where we seem to have gr
from a small mom-and-pop outlet to a microbiology super- Tesfaye Belay
store, a great number of people have supplied ideas, cri- Bluefield State College

Eric L. Buckles
Igoe, Sonya V ementioned Dillard University
Ray Burke. Of course, nothing happens in the lab without the
Erin A. Christensen
support of Mary Timmer
who has the ability to keep us all on track, supplied with Middlesex County College
what we need before we even know we need it. Finally, Iris M. Cook
it would seem karmically unwise not to thank Dr. Dave
Westchester Community College
Douglass, who has made sure the microbiology program has
r Lauren Cross
haven’t mentioned it, thanks for the new microscopes!
Wor-Wic Community College
Of course saying that one person “wrote” a book glosses
over the contributions of the many people who had a hand Natasha Dean
La Sierra University
a file in my computer. My deepest appreciation to sponsor-
ing editor L eithaupt, developmental editor Darlene Valencia Community College
Schueller, project manager Lisa Br
manager Amy Reed. Enjoyed the photos? Thank Lori Han- Denise Ferguson
Carolinas College of Health Sciences
ds, all of these people
had a hand in cr Robert Gessner
Lastly, there are thr Valencia Community College
than I could ever deliver e
pr ector for Carl Hamby
several of the photos you’ll see in the pages that follow while New York Medical College
my wife took on single par
ont por Daniece Harris-Williams
Hinds Community College Rankin Campus
at all hours of the day and night, and did so with good cheer,
fer encourage- Julie A. Huggins
ment, and when to of Arkansas State University

Tarrant County College

Reviewers James Masuoka
Midwestern State University
evision process, I was lucky enough to
obi-
ologists, who provided their take on the content, procedures, Long Island University – C.W. Post Campus
depth, and even order of the exer
have helped to make this a better book than I could ever Murad M. Odeh
have written on my own, and I thank them wholeheartedly. South Texas College
 Preface xi

Connie Pitman To the Student

University of Colorado at Colorado Springs

Narayanan Rajendran As an introductory student in microbiology, you may find

that the reasons behind a particular exercise appear overly
Kentucky State University
complex. Such is the nature of science, but the reasons
Lois Sealy should, at the very least, be apparent. The first step in clos-
Valencia Community College
understand, always, how each step relates to the overall
Peter Sheridan objective. It is just as important to understand why you are
Idaho State University doing something as it is to understand what it is you are
doing. If you can master both the why and the what, then
Jane Slone your success in microbiology will be assured.
Cedar Valley College/ DCCCD This book was written with you in mind. In other words,
everything in this book has been written to support some-
Janice Yoder Smith thing. That means that the introductory material helps to
Tarrant County College Northwest Campus explain the case study, the photos and diagrams are used to
clarify procedures, the glossary contains definitions of
Gabriel Swenson
microbiological terms, and websites are provided if you
Paine College would like further information on a topic. When you are
Stephen Wagner using this book, please, use this book. If the meaning of a
sentence is unclear, look to the accompanying figure; if a
Stephen F. Austin State University
word is a mystery, use the glossary; if space is provided for
Van Wheat a detailed drawing, give it your best shot—it will all be
South Texas College important soon. A well-used book becomes weathered as
om the book to the reader, and a lab
Dean E. Willis book is no different in this regard. Dog-eared pages, draw-
University of North Florida ings, notes, and circled definitions are all part of learning,
and the physical process of making the book yours parallels
Robin Woodard the intellectual process of making the information yours.
University of Virginia This is as true with microbiology as it is with any other
interest, job, or hobby. Take the steps to own the book, and
Linda Young you’ll own the information within.
Ohio Northern University

Jianmin Zhong
Humboldt State University

Brenda Zink
Northeastern Junior College
About the Author
Barry Chess has been a microbiologist at Pasadena City College since 1996. He received
his Bachelors and Masters degrees from California State University, Los Angeles, and

where his research centered on the expression of genes involved in the development of
muscle and bone.
At Pasadena City College, Barry developed a new course in human genetics and
helped to found a biotechnology program at the campus. He regularly teaches courses
in microbiology, biology
pendent research projects in biology and microbiology. Over the past several years
s interests have begun to focus on innovative methods of teaching that lead to
greater student understanding. He has written and reviewed cases for the National
Center for Case Study Teaching in Science and presented papers and talks on the use
of case studies in the classroom. He is a coauthor of the microbiology textbook Founda-
tions in Microbiology, having recently joined Kathy Talaro on the project.
Barry is a member of the American Society for Microbiology and regularly attends
meetings in his fields of interest, both to keep current of changes in the discipline and
to exchange teaching and learning strategies with others in the field.

xii
 C ASE STU DY E XERCISE
1
Safety Considerations in the Microbiology Laboratory

Escherichia coli O157:H7. J. Clini. Microbiol. 43:

2938–2939.
, Philadelphia, 2004. Emerging Infect. Dis. 12: 134–137.
e. Los Angeles Times.

to infection in the laboratory, and it is not an exaggeration

to say that proper safety procedures can be a matter of life
and death.
Safe laboratory procedures revolve around contain-
ment of microorganisms. Primary containment concerns
the protection of personnel and the laboratory environment
from exposure to infectious microbes. Proper microbiologi-
cal techniques, such as the safe transport and disposal of
cultures, along with the correct use of personal safety
equipment (e.g., gloves and safety glasses) go a long way
toward accomplishing the goal of personal containment.
deals with protecting the outside
environment from exposure to infectious organisms and
INTRODUCTION depends principally on the design of the laboratory and the
availability of equipment. There is generally little that can
The micr esents a number of unique be done to influence the physical aspects of the laboratory
challenges. Not only are the normal hazards of a laboratory other than not disabling safety features, such as keeping
envir e) open a door that should remain closed, turning off an
present, but so too ar ganisms. In fact, the exhaust fan, or removing a fire extinguisher.
microbiology lab is devoted to gr
very organisms that may cause us such harm! As the cases e mandated. W
above illustrate, laboratory workers are by no means immune deadly viruses obviously requires a gr
1
2 Case Study Exercise 1 Safety Considerations in the Microbiology Laboratory

e nonpathogenic. T • e the safety equipment is in the lab. Note

o- the location of the eye wash, safety shower, fire
or extinguisher, and first aid kit. Take a moment to learn
their operation; remember, if you need to use the eye
wash, you very well may not be able to see at the time!
regar At one
end, BSL-1 or
healthy person and requir During the Lab
BSL-4 or
• Always wear a lab coat while in the lab. Although you
may not be working yourself, another person in the lab
. Table 1.1 ecom- could have an accident. This garment should only be
used during lab and should remain in the lab. Even
The vast majority of introductory microbiology labora- discounting potential biohazards, a lab coat will protect
tories ar ganisms, e is a reason many of the chemicals
and the rules that apply in these laboratories have a very you will be working with are called stains.
common sense and feel about them. Your instructor may • W
eg- before leaving as well. Also wash when removing gloves
ulations as well as the organisms you are likely to work with and if you feel you may have contaminated yourself. If
as part of your course. Adhering to these guidelines will your laboratory sink has a hands-free method of
help to ensur obiology lab.
• Tie back any long hair, it is both a source of contamina-
tion and a fire hazard.
Prior to the Lab • ysol, or 10%
• Dress appropriately for the lab. No open toed shoes or e leaving
sandals. Clothing with baggy sleeves that could catch . If time permits, allow the disinfectant to
fire or hinder your movements should be avoided.

TABLE 1.1
 Case Study Exercise 1 Safety Considerations in the Microbiology Laboratory 3

• e book

will not consume precious bench space and where

they will be less likely to be contaminated by an
inadvertent spill.
• Nothing should go into your mouth while you are in
the laboratory. Do not smoke, eat, or drink in the lab,
even if no work is being done at the time.
• epresent a common portal of entry for
pathogens. Do not apply makeup, and never handle

• Organize your workplace before beginning (Figure 1.1).

Store culture tubes upright in a rack, never on their
side. Caps on tubes are generally not liquid tight, and
liquid will leak out (even fr e), leading

• The open flame produced by a Bunsen burner presents

an obvious danger in the laboratory
set up away from overhead equipment or shelving and
the immediate area should be free of combustible
materials such as notes or books. Prior to lighting the
burner, quickly inspect the hose for holes, cracks, or
leaks, and be sure it fits securely on both the gas valve Figure 1.2 Autoclaves use steam under pressure to sterilize
and the burner. biohazardous materials prior to disposal.

Disposal of Contaminated Materials

disposal of lab materials depends on whether or not it will
Most material in the microbiology laboratory must be decon-
be reused. In any event, the contents of plates or tubes
taminated prior to being disposed of or reused, and this is
should never be touched by hand.
most often accomplished using an autoclave (Figure 1.2),
which uses steam under high pressure to kill even the most •
resistant organisms. e tubes, gloves, and similar nonreusable items in the biohazard
glass pipettes, and the like are washed and reused. Plastic bag (Figure 1.3a
Petri dishes melt during the decontamination process and but there is no need to remove labels or tape from
are discar items.
such as tongue depressors, needles, and swabs. In general, • Reusable supplies such as culture tubes and glass
pipettes should have all labels removed and placed in a

• Used micr

e being discarded.

Safety Considerations
• Be r

essant
drugs, or being pregnant should be candidly discussed
.
• Always wear gloves when handling blood or blood
products. Blood-borne pathogens have special
procedur
should only be done with the explicit knowledge of
Figure 1.1 uctor.
of a lab coat, gloves, and eye protection. Also note that long hair is • W
tied back and the work area is free of clutter. microorganisms as a result of a spill.
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Order Blank
 SALES BOOK
9. The sales book used in this set exhibits some features not heretofore
shown. At the right are three columns for the distribution of sales. At the
left, in addition to columns for number, date, and folio, are two columns
headed cash and sales ledger. All cash sales are entered in the cash column,
and all sales on account are entered in the sales ledger column. At the end
of the week or month the total of the sales ledger column is posted to the
debit of the sales ledger account in the general ledger, while the totals of the
sales columns at the right are posted to the credit of the sales account in the
general ledger.
 INVOICE REGISTER
10. A form of purchase book, which also combines an invoice register, is
shown in this set. Unlike the forms of purchase book with which the student
has been made familiar, this invoice register gives full particulars as to terms,
discount, when due, and when and how paid.
The combined footings of the two department columns must of course agree
with the footing of the amount column. At the end of the month the total of
the amount column is posted to the credit of purchase ledger account in the
general ledger, and the totals of the department columns are posted to the
debit of the purchase account in the general ledger.
The details of payment are kept in the invoice register as a memorandum
only. This provides a convenient record of unpaid invoices, showing when
each is due.
 CASH BOOK
11. In this set we introduce a columnar cash book which also serves as a
journal for cash transaction and is known as a cash journal. The principal
advantage of a columnar book lies in the opportunity to introduce columns
with special headings for accounts to which entries are frequent. Not only
does this permit of carrying footings to the end of the month with one
posting to the ledger account, but it provides a convenient classification of
receipts and expenditures with a complete segregation of items of a given
class.
In the form illustrated, columns are provided on the debit side for cash,
purchase ledger (subdivided for discount and amount), bank deposits, and
sundries; on the credit side, cash, sales ledger (subdivided for discount and
amount), cash sales, bank withdrawals, and sundries. At first glance it might
appear that this form is a departure from the regular form of cash book, but
it should be remembered that the cash columns are the only ones having
anything to do with the cash account. A cash receipt is entered in the cash
debit column, but the amount is credited to its source through the proper
credit column; thus a payment received on account is debited to cash and
credited through the sales ledger column. A deposit is credited to cash and
debited to bank deposits; the payment of a purchase ledger account by
check is credited to bank withdrawals, and debited to purchase ledger
account.
The discount columns are memorandum columns only, the net cash being
entered in the amount columns under purchase and sales ledger. These
columns are included that the total payment may be posted to personal
accounts in purchase or sales ledger. The totals of these columns are to be
posted to discount and interest columns at the end of the month.
The total amount to be posted to the debit of the purchase ledger account
and to the credit of sales ledger account is made up of the totals of the
discount and amount columns.
The sundries columns are provided for all entries for which there are no
special columns and are used principally for transactions affecting general
ledger accounts. These columns are sometimes used for ordinary journal
entries not involving an exchange of cash, but their use for this purpose is
strongly advised against. The cash book should be used exclusively for
recording cash transactions. When columnar purchase, sales, and cash books
are used, the journal is only needed for adjusting and closing entries, and for
this purpose it is best to provide an ordinary two-column journal.
 SUBDIVISION OF EXPENSE ACCOUNT
12. In every business there are several classes of expense and it is very
useful to know the exact amount of each class. When all expenses are
charged under one head, it is impossible to determine without considerable
checking, whether or not any particular class of expense is more than it
should be. It is customary, therefore, to subdivide expense and to open
accounts in the ledger for different classes of expense. Some subdivisions in
common use are rent (paid), insurance, taxes, interest and discount, in
freight, out freight, salaries, labor, fuel, office supplies, telegraph and
telephone, postage, general expense, etc. The exact subdivisions used must
of necessity be governed by the nature of the business. For instance, the
item of telegraph and telephone charges may be of importance in one
business, while in another, the number of such charges would be so small
that a separate account is not warranted.
 PETTY CASH VOUCHER
13. A form of envelope voucher for petty cash is illustrated. A strong manilla
envelope in what is known as size #10 takes the place of the petty cash
book. It is ruled for a record of payments, and a receipt for each payment is
placed in the envelope. At the bottom is a space for a distribution of the
amounts to the proper accounts. When the petty cash fund is depleted—or
at stated intervals—a check is drawn for the amount expended and it is
charged through the cash book, leaving petty cash intact. The amount of the
petty cash fund is considered as cash on hand, and the voucher envelope
accounts for any part of the fund not actually in the cash drawer. Petty cash
should be used sparingly, as it is intended only for small expense items when
it is inconvenient to give a check. When the books are closed, the petty cash
expenditures may be charged through the cash book as cash payments,
instead of drawing a check.
Petty Cash Voucher
 TREATMENT OF PROTESTED PAPER
14. When a note, draft, or check is protested, the bank will charge us with
the protest fee in addition to the face of the paper. The total amount must
then be charged to the one from whom the paper was received. Suppose the
check of Jones & Laughlin for $100.00 goes to protest and is returned to us
with a protest fee of $2.50—the entry will be:
Jones & Laughlin $102.50
Bank $102.50
Check No. 16 given to First Nat.
Bank to cover J. & L. check for
$100.00, protest fee $2.50.
 SAMPLE TRANSACTIONS
15. D. A. Hall is engaged in the business of a wholesale dealer in men's and
boys' clothing. On Feb. 1st. his balance sheet is as follows:
Balance Sheet, Feb. 1st, 1909.
Assets
Cash
In Bank $1,765.20
In Office 125.00

Total Cash $1,890.20

Accounts and Bills Receivable

Bills Receivable 850.00
Henry James, Due
Feb. 5 350.00
David Traver & Co.,
Due
Feb. 15 500.00
Accounts Receivable 1,124.00
Frank Weitz 234.00
John Gorham 150.00
George Golden 300.00
Clayton & Co. 275.00
Henry Ames 165.00

Total Accounts and Bills 1,974.00

Rec.

Inventory
Men's clothing $2,240.00
Boys' " 1,200.00
 Total Inventory $3,440.00

Total Assets $7,304.20

Liabilities
Accounts and Bills Payable
Bills Payable 650.00
Henry Weir & Co., Due
Feb. 7 450.00
A. Stein & Co., Due
March 1 200.00

Accounts Payable 675.00

D. Meyer & Bro. 150.00

Altman & Sons 350.00
Garson & Co. 175.00

Total Accounts and

Bills
Payable 1,325.00
Total Liabilities 1,325.00

Present Worth 5,979.20

The following transactions are entered on the books:
—Feb. 1st—
Sold to D. A. Marcus & Son
10 overcoats 7.50 75.00
10 men's suits 6.75 67.50 142.50

—1st—
Sold to H. A. Branch
15 boys' suits 3.50 52.50
 —1st—
Rec'd from Geo. Golden
Cash on account 150.00

—2nd—
Deposited in 1st Nat. Bank 150.00

—2nd—
Sold to John Gorham
10 men's suits 7.00 70.00
10 men's suits 6.50 65.00 $135.00

—2nd—
Sold to Larson & Anderson
5 boys' suits 2.75 13.75
10 men's suits 6.50 65.00 78.75

—2nd—
Sold for cash
Men's clothing 37.50

—3rd—
Sold to Tallman & Co.
15 men's overcoats 7.25 108.75

—3rd—
Paid 1 month's rent, Ck.
No. 1 75.00

—3rd—
Received from John Gorham
Cash on account 150.00
 —4th—
Bought from Carson & Scott
36 men's corduroy coats 3.00 108.00
12 men's corduroy coats 3.50 42.00 150.00
Terms 2/10, 1/30, n/60

—4th—
Deposited in 1st Nat. Bank 187.50

—4th—
Sold to Harris & Rogers
12 men's corduroy coats 3.75 45.00
5 overcoats 8.00 40.00 85.00

—4th—
Received from Frank Weitz
Note at 30 days, 6% 234.00

—5th—
Sent to D. Meyer & Bro. Ck.
No. 2 $150.00

—5th—
Received from Henry James
Cash to apply on note 200.00
Cash for interest 1.75 $201.75
New note 30 days, 6% 150.00

—5th—
Bought from Adler & Co.
50 men's suits 6.25 312.50
Terms 3/10, 1/30, n/60

—5th—
Paid salesman's salary Ck.
No. 3 25.00

—5th—
Drew for personal use Ck.
No. 4 50.00

—7th—
Sold to Henry Ames
20 men's suits 7.50 150.00
10 boy's suits 2.75 27.50 177.50

—7th—
Sold to Ackley & Son
10 boy's overcoats 3.00 30.00
10 boy's suits 2.75 27.50 57.50

—7th—
Received from Clayton & Co.
Check to apply on acct. 200.00

—7th—
Deposited in 1st Nat. Bank 401.75

—7th—
Paid our note to H. Weir & Co.
Check No. 5 450.00

—8th—
Sold to H. J. Andrews
10 men's overcoats 8.00 80.00
12 men's corduroy coats 4.50 54.00 $134.00

—8th—
Paid express on shipment from
Carson & Scott, Ck. No. 6 .90

—8th—
Received from Henry Ames
Cash on account 165.00

—8th—
Sold for cash
1 job lot boy's clothing 87.50

—9th—
Deposited in 1st Nat. Bank 252.50

—9th—
Received from bank, check of
Clayton & Co., protested
for non-payment.
Amount of check 200.00
Protest fees 2.50

—10th—
Sold to Harris & Landis
10 men's overcoats 7.75 77.50

—10th—
Sold to Frank Weitz
12 men's corduroy coats 3.75 45.00

—10th—
Paid Carson & Scott
Check No. 7 147.00
Discount 2% 3.00
 —10th—
Paid electric light bill
Check No. 8 3.75

—10th—
Received from Clayton & Co.
Cash to redeem protested
check $202.50

—10th—
Inventory at close of business,
Feb. 10
Men's clothing 1,898.75
Boy's clothing 1,247.75
Journal entries are to be made to get the accounts, as shown on Feb. 1st,
recorded on the books. The transactions are properly entered in journal,
cash book, sales book, and invoice register, and posted to ledger. The
accounts in the general ledger are closed into trading, and profit and loss—
the net profit is credited to proprietor's account—a trial balance is taken after
the ledger is closed, and a balance sheet is made. Statements are prepared
from sales and purchase ledgers, which agree with the balances of their
controlling accounts. All accounts in the general ledger are properly ruled
and balances carried forward.
Opening Entry in Journal
Adjusting Journal Entries
Sales Book and Invoice Register
SCENE IN SOUTH WATER STREET, WHERE THE COMMISSION HOUSES OF
CHICAGO'S GREAT PRODUCE MARKETS ARE LOCATED

Columnar Cash Journal

Columnar Cash Journal
Sales Ledger
Sales Ledger
Sales and Purchase Ledgers
Purchase and General Ledgers
General Ledger
General Ledger
General Ledger
Statement of Sales and Purchase Ledger
Balance Sheet and Trial Balance of General Ledger
 EXERCISES
16. The following transactions are recorded on the books of Parker and
Hoadley, Omaha, Neb., wholesale dealers in tea and coffee. In recording
these transactions use is made of the books and forms illustrated in this
section.

January 2nd, 1909

A partnership is formed on this date between K. J. Parker and D. C.

Hoadley for the purpose of conducting a wholesale tea and coffee
business, in the name of Parker & Hoadley, the principal place of
business to be Omaha, Neb. Parker invests $3,000.00 cash. Hoadley
invests $2,000.00 cash. It is agreed that profits are to be shared on the
basis of capital invested, capital to draw interest at 6%, and interest at
6% to be paid on withdrawals. The books are to be closed monthly and
the profits divided between the partners. Hoadley is to assume the
entire responsibility for the conduct and management of the business,
for which he is to receive a salary of $150.00 per month, payable in
installments of $75.00 on the 15th and 31st of each month.
Deposited in Western National Bank $5,000.00.
Withdrew from bank, Ck. No. 1 petty cash $25.00.
Bought from Leggitt & Co., New York, 30 chests Japan tea, 1,455# at
.37½, 20 chests Oolong tea, 972# at .40; terms net 30, 2/10, f. o. b. N.
Y.
Bought from Laughlin & Co., Chicago, 20 sacks Rio coffee, 1,020# at
.22½, 20 sacks Java coffee, 985# at .25; 20 sacks Mocha coffee, 970#
at .25; terms net 30, 2/10, f. o. b. Omaha.
Paid rent of store 1 month to James Roberts, Ck. No. 2, $60.00.

—3rd—

Sold to Ames & Johnson, 92 12th St., on account, 3 chests Japan tea,
149# at .48; sack Rio coffee, 50# at .28; 1 sack Java coffee, 52# at .32.
Sold to Landis & Snow, So. Omaha, on account, 2 chests Oolong tea,
101# at .52; 1 sack Mocha coffee, 47# at .32; 2 sacks Rio coffee, 98#
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