| 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.
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| 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).
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| 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).
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| 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.)
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5.2
5.3 | - Importance of growth factors with use of siderophores as an example.
- Phenotypic variability according to environment.
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| 5.4 | - The bacterial growth curve.
| - Growth curves – explanation of graphing theory and formulas beyond what is in manual.
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| 6 | | |
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.
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8.1
8.2 | - Mutation – detecting antibiotic-resistant mutants.
- Recombination – "mating" two auxotrophs to form recombinants that resemble prototrophs.
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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.
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| 10.1 | - The antibiotic disc sensitivity test.
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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).
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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.
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| 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).
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| 17 | - The last great unknown – a review of isolation and identification with construction of a relevant dichotomous key.
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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.
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