Bacteriology 102:
Lab Manual Contents
and Topics

YOU ARE HERE:
John L's Bacteriology Pages >
Bact.102 Website–Fall 2006 >
Lab Manual Contents & Topics
Page Last
Modified:
8/6/03

The lab manual referred to herein is referenced here.


This page undergoes frequent fine-tuning and might still be a bit "rough" but hopefully useful nonetheless.

Here is a list of related items (click on the X):

  • X  General outline of topics covered in this course.
  • X  Some questions to think about!
  • X  A list of important terms and concepts which may not be explained well in the manual.
  • X  Our older "lab manual index" of commonly-referred-to items (which is also on the syllabus with the course introduction, schedule and point system).
  • X  Places where a lot of us tend to backslide!

Following are the Table of Contents of the Manual followed by a General List of Topics usable as a study guide with links to various web pages.


EXPERIMENTS

  • General introduction to the laboratory

  • Rules for laboratory cleanup

  • 1.  MICROORGANISMS IN THE ENVIRONMENT

  • 2.  MICROSCOPIC OBSERVATIONS OF BACTERIA
    • Results sheet for gram stain unknown

  • 3.  BASIC PURE CULTURE TECHNIQUES

  • 4.  QUANTITATIVE MICROBIOLOGY

  • 5.  SELECTED ASPECTS OF BACTERIAL GROWTH AND NUTRITION
    • 5.1  OXYGEN RELATIONSHIPS AND RELATED THINGS
    • 5.2  EXAMPLE OF A GROWTH FACTOR REQUIREMENT
    • 5.3  EXAMPLE OF PHENOTYPIC VARIATION
    • 5.4  THE BACTERIAL GROWTH CURVE

  • 6.  BACTERIAL MOTILITY
    • Results sheet for motility unknown

  • 7.  CHARACTERIZATION, DIFFERENTIATION AND IDENTIFICATION OF BACTERIA
    • 7.1  CHARACTERIZATION OF TWELVE KNOWN BACTERIA
    • 7.2  CHARACTERIZATION AND IDENTIFICATION OF TWO UNKNOWN CULTURES  
    • Appendix 7A.  Morphology and arrangement of bacterial cells
    • Appendix 7B.  Summary of biochemical tests in Experiment 7
    • Results sheet for mixed unknowns

  • 8.  AN INTRODUCTION TO BACTERIAL GENETICS
    • 8.1  SELECTION OF MUTANTS
    • 8.2  CONJUGATION AND GENETIC RECOMBINATION

  • 9.  BACTERIOPHAGES, THE VIRUSES OF BACTERIA
    • 9.1  PHAGE ASSAY
    • 9.2  PHAGE-TYPING
    • 9.3  DETECTION OF BACTERIOPHAGES IN THE ENVIRONMENT

  • 10.  INHIBITION AND KILLING OF BACTERIA
    • 10.1  THE ANTIBIOTIC DISC-SENSITIVITY TEST
    • 10.2  ISOLATION OF STREPTOMYCES

  • 11.  ISOLATION OF BACTERIA FROM NATURAL SOURCES
    • 11.1  ENRICHMENT AND ISOLATION OF ANOXYGENIC PHOTOTROPHS
    • 11.2  ISOLATION OF BACILLUS
    • 11.3  ENRICHMENT AND ISOLATION OF NITROGEN-FIXING BACTERIA
    • Appendix 11A.  Guidelines for preparing formal laboratory reports and posters

  • 12.  LACTIC ACID BACTERIA AND AN INTRODUCTION TO FOOD MICROBIOLOGY

  • 13.  SELECTED GENERA OF MEDICAL IMPORTANCE
    • 13.1  STAPHYLOCOCCUS, STREPTOCOCCUS AND NEISSERIA
    • 13.2  OTHER PATHOGENIC BACTERIA OF INTEREST

  • 14.  THE ENTERIC BACTERIA (THE FAMILY ENTEROBACTERIACEAE)
    • 14.1  ISOLATION AND IDENTIFICATION OF TWO ENTERIC "UNKNOWNS"
    • 14.2  SEROLOGICAL IDENTIFICATION OF A SALMONELLA CULTURE
    • Appendix 14A.  Outline of plating and preliminary screening media used for enterics
    • Appendix 14B.  Key for enteric unknowns
    • Results sheet for enteric unknowns

  • 15.  THE BACTERIOLOGICAL EXAMINATION OF WATER
    • Appendix 15A.  Using the MPN technique to estimate microbial concentration
    • Water bacteriology data sheet

  • 16.  PATHOGENICITY AND VIRULENCE: KOCH'S POSTULATES
    • 16.1  SOFT ROT OF POTATOES
    • 16.2  WILT OF TOMATO PLANTS

  • 17.  THE FINAL UNKNOWN IDENTIFICATION
    • Results sheet for unknowns

APPENDICES:  GENERAL

  • APPENDIX A.  USE OF THE MICROSCOPE

  • APPENDIX B.  INTRODUCTION TO PURE CULTURE TECHNIQUES
    • B.1  PROPER STERILIZATION OF LOOPS AND NEEDLES
    • B.2  PREPARATION OF A BACTERIAL SMEAR FOR STAINING
    • B.3  TUBE-TO-TUBE TRANSFERS
    • B.4  PLATE-TO-TUBE TRANSFERS
    • B.5  STREAK PLATES
    • B.6  USE OF PIPETTORS AND PREPARATION OF SPREAD PLATES
    • B.7  PREPARATION OF POUR PLATES

  • APPENDIX C.  INTRODUCTION TO DILUTION THEORY

  • APPENDIX D.  NUTRITION AND CULTIVATION OF MICROORGANISMS
    • D.1  MEETING THE NUTRITIONAL NEEDS OF MICROORGANISMS
    • D.2  STERILIZATION OF CULTURE MEDIA AND ASSOCIATED ITEMS

  • APPENDIX E.  SUMMARY OF CULTURE MEDIA
    • E.1  SOME COMMONLY-USED CONSTITUENTS OF MICROBIOLOGICAL MEDIA
    • E.2  FORMULATIONS OF MEDIA USED IN THIS MANUAL

  • APPENDIX F.  SUMMARY OF SELECTED BIOCHEMICAL TESTS

  • APPENDIX G.  STAINING OF BACTERIA FOR MICROSCOPIC EXAMINATION
    • G.1  THE SIMPLE STAIN
    • G.2  THE GRAM STAIN
    • G.3  THE CAPSULE STAIN
    • G.4  THE ENDOSPORE STAIN
    • G.5  THE ACID-FAST STAIN

  • APPENDIX H.  MISCELLANEOUS ITEMS
    • H.1  PROPER USAGE OF TERMS AND SCIENTIFIC NAMES
    • H.2  DICHOTOMOUS KEYS AND FLOW CHARTS

APPENDICES:  HELPFUL REVIEW MATERIAL

  • APPENDIX W.  LIST OF BACTERIOLOGICAL TERMS TO KNOW

  • APPENDIX X.  SELECTED QUESTIONS FROM OLD QUIZZES

  • APPENDIX Y.  SAMPLE DILUTION PROBLEMS

  • APPENDIX Z.  EXAMPLE OF A FINAL EXAMINATION

 

Exp.# Major Items and Concepts Explanatory Material in the Lab Manual Appendices and on the Web
1
2
  • Concept of cell vs. CFU vs. colony.
  • Shapes and arrangements of cells.
  • Environmental sampling (swabbing plates).
  • Elementary bacterial quantitation (plate counts of soil and water samples).
  • Preparation of smears and wet mounts.
  • Simple, gram, capsule and acid-fast stains.
  • Use of regular and phase-contrast microscopes.
3
  • Aseptic transfer techniques involving loops, needles, tubes and plates.
  • Some basic techniques.
  • Appendix B (basic transfer techniques, streak-plating).
  • Appendix F (Glucose Fermentation Broth).
4
  • Dilution-plating of hamburger.
  • Dilution Theory:  Determining the "total aerobic" and gram-negative plate counts (with use of formulas to obtain CFU/ml).
  • Appendix B (use of pipettors, Vortex machines and hockey sticks with dilution-making and spread-plating).
  • Appendix C (all one would ever need to know about interpreting results from dilution-plating).
  • More dilution theory (explaining things in App. C a better way).
  • General practice problems on pages 122, 168 and 169 – solutions are here.
  • MacConkey Agar – first example of a selective-differential medium (also on page 126).
5.1
  • Basic Catabolism and Oxygen Relationships:  Connecting growth responses in the oxygen relationship test to aerobic respiration and fermentation; understanding the narrow focus and limitations of the concept; correlation of results with other tests such as the catalase test.
  • The use and value of Glucose O/F Medium.
  • (Consideration of anaerobic respiration and anoxygenic phototrophy come later in Exps. 7 and 11.1, respectively.)
5.2
5.3
  • Importance of growth factors with use of siderophores as an example.
  • Phenotypic variability according to environment.
 
5.4
  • The bacterial growth curve.
  • Growth curves – explanation of graphing theory and formulas beyond what is in manual.
6
  • Bacterial motility.
7.1
7.2
  • Bacterial Identification:  Comparative bacterial morphology and physiology and the use of phenotypic tests.
  • Introduction to extracellular enzymes (with starch hydrolysis) and anaerobic respiration (with nitrate reduction).
  • Generation of a database (on a table) for the identification of unknowns.
  • Brief discussion of dichotomous keys and flow charts.
  • Brief discussion of differentiation and identification by genotypic tests; construction and meaning of phylogenetic trees.
8.1
8.2
  • Mutation – detecting antibiotic-resistant mutants.
  • Recombination – "mating" two auxotrophs to form recombinants that resemble prototrophs.
9.1
9.2
  • Bacteriophages – quantitation and the concept of "plaque-forming units"; specificity and the technique of "phage-typing."
  • Sample phage quantitation problem in Appendix Y; solution is here.
10.1
  • The antibiotic disc sensitivity test.
 
11.1
11.2
11.3
10.2
9.3
  • Enrichment and Isolation of Microorganisms – general principles.
  • Taking and tabulating notes on procedures and observations; more about flow charts and tables.
  • Preparation of formal reports and posters.
  • Enrichment and isolation of purple non-sulfur photosynthetic bacteria; testing isolates to confirm anoxygenic phototrophy (Exp. 11.1).
  • Isolation of Bacillus (Exp. 11.2).
  • Enrichment and isolation of N2-fixing bacteria; testing isolates to confirm nitrogen-fixing ability (Exp. 11.3).
  • Isolation of Streptomyces; testing for antibiotic production (Exp. 10.2).
  • Isolation of bacteriophages (Exp. 9.3).
12
13.1
14.1
14.2
  • The use of dichotomous keys.
  • The lactic acid bacteria – including selective isolation; using starter cultures and wild fermentations in the food industry (with yogurt and sauerkraut as examples).
  • Selected cocci of medical relevance – Staphylococcus, Streptococcus (and Enterococcus), and Neisseria – general characteristics and specific tests such as oxidase, coagulase and hemolysis.
  • The enteric bacteria – selective enrichment and isolation; phenotypic vs. genotypic tests with more about phylogenetic trees; demonstration of the API-20E system; use of serology in Salmonella identification.
  • Differential Media:  "Programming" isolation and test media to differentiate certain physiologial types of bacteria from others.
15
  • Water analysis – "total aerobic plate count" revisited, plus the enrichment, detection, enumeration, isolation and identification of coliforms.
  • Dilution Theory:  Use of the most-probable number (MPN) method to enumerate certain types of bacteria such as coliforms.
  • The concept of indicator organisms – in general and specifically regarding water quality (coliforms).
17
  • The last great unknown – a review of isolation and identification with construction of a relevant dichotomous key.
 

Additional items in Appendices and/or on Web:

  • Appendix E.2: Formulations (not to memorize!) and brief information about culture media used in the manual.
  • Appendix F: Selected biochemical tests.
  • Appendix H: Proper usage of terms and scientific names (p. 149) – reproduced here.

  • Appendix W: List of terms to know for each experiment.
  • Terms and concepts not defined too well in the manual are listed here.
  • Appendix X: Sample questions from earlier quizzes. Answers are here.
  • Appendix Z: An old lab exam. Answers are here.
  • More questions to think about.
  • Site Outline of related pages.

GO TO:
Home Page of Bacteriology 102 Web Site
Index of the General Bacteriology Pages  

Page last modified on 8/6/03 at 3:30 PM.

John Lindquist, Department of Bacteriology

University of Wisconsin – Madison