Kleyn−Bicknell: Front Matter Preface © The McGraw−Hill Microbiology Experiments: Companies, 2003 A Health Science Perspective, 4/e Preface To the Student ten to use as little expensive media and equipment as possible. The manual contains more exercises A microbiology laboratory is valuable because it ac- than can be done in one course so that instructors tually gives you a chance to see and study microor- will have an opportunity to select the appropriate ganisms firsthand. In addition, it provides you with exercises for their particular students and class. We the opportunity to learn the special techniques hope that the instructors find these laboratories an used to study and identify these organisms. The enjoyable component of teaching microbiology. ability to make observations, record data, and ana- lyze results is useful throughout life. It is very important to read the scheduled exer- Acknowledgments cises before coming to class, so that class time can We would like to acknowledge the contributions of be used efficiently. It is helpful to ask yourself the the lecturers in the Department of Microbiology at purpose of each step as you are reading and carrying the University of Washington who have thought- out the steps of the experiment. Sometimes it will fully honed laboratory exercises over the years until be necessary to read an exercise several times be- they really work. These include Dorothy Cramer, fore it makes sense. Carol Laxson, Mona Memmer, Janis Fulton, and Conducting experiments in microbiology labora- Mark Chandler. Special thanks to Dale Parkhurst tories is particularly gratifying because the results for his expert knowledge of media. We also thank can be seen in a day or two (as opposed, for instance, the staff of the University of Washington media to plant genetics laboratories). Opening the incuba- room for their expertise and unstinting support. tor door to see how your cultures have grown and We also want to thank Eugene and Martha how the experiment has turned out is a pleasurable Nester, Nancy Pearsall, Denise Anderson and moment. We hope you will enjoy your experience Evans Roberts for their text Microbiology: A Human with microorganisms as well as acquire skills and un- Perspective. This text was the source of much of the derstanding that will be valuable in the future. basic conceptual material and figures for our labo- ratory manual. And with great appreciation, many To the Instructor thanks to our editor, Deborah Allen, for her sugges- tions, assistance, and ever cheerful support. The manual includes a wide range of exercises— Additional thanks to Meridian Diagnostics in some more difficult and time-consuming than oth- Cincinnati for their generous offer to make diag- ers. Usually more than one exercise can be done in nostic kits available for some exercises. We also a two-hour laboratory period. In these classes, stu- thank the following instructors for their valuable dents can actually see the applications of the prin- input on the revision of this manual. ciples they have learned in the lectures and text. We have tried to integrate the manual with the Reviewers text Microbiology: A Human Perspective, Fourth Edition by Eugene Nester et al. Barbara Beck Kathleen C. Smith The exercises were chosen to give students an Rochester Community Emory University opportunity to learn new techniques and to expose and Technical College Evert Ting them to a variety of experiences and observations. Mark Chatfield Purdue University Calumet It was not assumed that the school or department West Virginia State College Robert Walters had a large budget, thus exercises have been writ- James Madison University vii Kleyn−Bicknell: Front Matter Laboratory Safety © The McGraw−Hill Microbiology Experiments: Companies, 2003 A Health Science Perspective, 4/e Laboratory Safety To be read by the student before beginning any lab- 9. Avoid creating aerosols by gently mixing oratory work. cultures. Clean off the loop in a sand jar 1. Do not eat, drink, smoke, or store food in the before flaming in the Bunsen burner. laboratory. Avoid all finger-to-mouth contact. 10. If a culture is dropped and broken, notify 2. Never pipette by mouth because of the danger the instructor. Cover the contaminated of ingesting microorganisms or toxic chemicals. area with a paper towel and pour disinfec- 3. Wear a laboratory coat while in the laboratory. tant over the material. After ten minutes, Remove it before leaving the room and store it put the material in a broken glass container in the laboratory until the end of the course.* to be autoclaved. 4. Wipe down the bench surface with disinfectant 11. Carefully follow the techniques of handling before and after each laboratory period. cultures as demonstrated by the instructor. 5. Tie long hair back to prevent it from catching 12. When the laboratory is in session, the doors fire in the Bunsen burner or contaminating and windows should be shut. A sign should be cultures. posted on the door indicating that it is a 6. Keep the workbench clear of any unnecessary microbiology laboratory. books or other items. Do not work on top of 13. Be sure you know the location of fire the manual because if spills occur, it cannot extinguishers, eyewash apparatus, and other be disinfected easily. safety equipment. 7. Be careful with the Bunsen burner. Make sure 14. Wash your hands with soap and water after that paper, alcohol, the gas hose, and your any possible contamination and at the end of microscope are not close to the flame. the laboratory period. 8. All contaminated material and cultures must 15. If you are immunocompromised for any reason be placed in the proper containers for (including pregnancy), it may be wise to autoclaving before disposal or washing. consult a physician before taking this class. * Other protective clothing includes closed shoes, gloves (optional), and eye protection. viii Kleyn−Bicknell: Front Matter Laboratory Safety © The McGraw−Hill Microbiology Experiments: Agreement Companies, 2003 A Health Science Perspective, 4/e Laboratory Safety Agreement To be read by the student before beginning any lab- 9. Avoid creating aerosols by gently mixing oratory work. cultures. Clean off the loop in a sand jar 1. Do not eat, drink, smoke, or store food in the before flaming in the Bunsen burner. laboratory. Avoid all finger-to-mouth contact. 10. If a culture is dropped and broken, notify 2. Never pipette by mouth because of the danger the instructor. Cover the contaminated of ingesting microorganisms or toxic chemicals. area with a paper towel and pour disinfec- 3. Wear a laboratory coat while in the laboratory. tant over the material. After ten minutes, Remove it before leaving the room and store it put the material in a broken glass container in the laboratory until the end of the course.* to be autoclaved. 4. Wipe down the bench surface with disinfectant 11. Carefully follow the techniques of handling before and after each laboratory period. cultures as demonstrated by the instructor. 5. Tie long hair back to prevent it from catching 12. When the laboratory is in session, the doors fire in the Bunsen burner or contaminating and windows should be shut. A sign should be cultures. posted on the door indicating that it is a 6. Keep the workbench clear of any unnecessary microbiology laboratory. books or other items. Do not work on top of 13. Be sure you know the location of fire the manual because if spills occur, it cannot extinguishers, eyewash apparatus, and other be disinfected easily. safety equipment. 7. Be careful with the Bunsen burner. Make sure 14. Wash your hands with soap and water after that paper, alcohol, the gas hose, and your any possible contamination and at the end of microscope are not close to the flame. the laboratory period. 8. All contaminated material and cultures must 15. If you are immunocompromised for any reason be placed in the proper containers for (including pregnancy), it may be wise to autoclaving before disposal or washing. consult a physician before taking this class. * Other protective clothing includes closed shoes, gloves (optional), and eye protection. I have read and understood the laboratory safety rules: __________________________________________________________ ______________________ Signature Date ix Kleyn−Bicknell: I. Basic Microbiology Introduction to © The McGraw−Hill Microbiology Experiments: Microbiology Companies, 2003 A Health Science Perspective, 4/e PART ONE BASIC MICROBIOLOGY I NTRODUCTION to Microbiology When you take a microbiology class, you have an breaking down dead plant and animal material into opportunity to explore an extremely small biologi- basic substances that can be used by other growing cal world that exists unseen in our own ordinary plants and animals. Photosynthetic bacteria are an world. Fortunately, we were born after the micro- important source of the earth’s supply of oxygen. scope was perfected so we can see these extremely Microorganisms also make major contributions in small organisms. the fields of antibiotic production, food and bever- A few of these many and varied organisms are age production as well as food preservation, and pathogens (capable of causing disease). Special more recently, recombinant DNA technology. The techniques have been developed to isolate and principles and techniques demonstrated here can identify them as well as to control or prevent their be applied to these fields as well as to medical tech- growth. The exercises in this manual will empha- nology, nursing, or patient care. This course is an size medical applications. The goal is to teach you introduction to the microbial world, and we hope basic techniques and concepts that will be useful to you will find it useful and interesting. you now or can be used as a foundation for addi- Note: The use of pathogenic organisms has been tional courses.