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General Microbiology II

Laboratory Exercises - Spring 2009

January 12 and 16

Introduction and Overview

Laboratory Safety

Microbial Microcosm (Exercise 1) - setup sulfate-enriched and nitrate-enriched Winogradsky columns

January 19 and 23

Martin Luther King Day Week - No Class

January 26 and 30

Microbial Microcosm (Exercise 1) - unwrap columns, examine column appearance

Selection (Exercise 2) - heat, dilute and plate bacterial mixture

Cyanobacteria (Exercise 3) - begin critical examination

February 2 and 6

Microbial Microcosm (Exercise 1) - examine sulfate-enriched columns for black spots

Selection (Exercise 2) - count colonies, analyze results, plan experiment

Cyanobacteria (Exercise 3) - complete critical examination

February 9 and 13

Microbial Microcosm (Exercise 1) - examine sulfate-enriched columns for black spots; examine nitrate-enriched columns

Selection (Exercise 2) - perform experiment

Fungi (Exercise 4) - begin critical examination

Notebook Assessment 1 (5 points)

February 16 and 20

Microbial Microcosm (Exercise 1) - isolate purple photosynthetic bacteria and inoculate Desulfovibrio agar tubes from sulfate-enriched columns; isolate and test nitrate-enriched column microbes

Selection (Exercise 2) - analyze and discuss results of experiments

Fungi (Exercise 4) - complete critical examination

February 23 and 27

Microbial Microcosm (Exercise 1) - examine sulfate-enriched columns for green and purple bacteria; examine Desulfovibrio agar tubes, analyze and discuss microcosm results

Bacterial Synergism (Exercise 5) - streak plates to set uo experiment

Succession in Pond Water (Exercise 6) - examine day 1 slides, setup for day 4

Report 1 Due (15 points ... Selection - Exercise 2)

March 2 and 6

Microbial Microcosm (Exercise 1) - examine sulfate-enriched columns for green and purple bacteria; examine Desulfovibrio agar tubes, analyze and discuss microcosm results

Bacterial Synergism (Exercise 5) - analyze and discuss results

Succession in Pond Water (Exercise 6) - examine day 4 slides, analyze and discuss results

March 9 and 13

Spring Break - No Class

March 18 (Wednesday evening)

Examination 1 (30 points ... LaboratorySafety and Exercises 1-6)

March 23 and 27

Skin and Respiratory Tract Microbiota (Exercise 7) - inoculate plates

Microscopic Examination of Leukocytes (Exercise 8) - critical examination

March 30 and April 3

Skin and Respiratory Tract Microbiota (Exercise 7) - characterize bacteria

Functions of Antibodies (Exercise 9) - bacterial agglutination

Antibiotic Sensitivity (Exercise 10) - inoculate plates and add antibiotic discs

April 6 and 10

Antibiotic Sensitivity (Exercise 10) - analyze and discuss results

Transmission of Microbes (Exercise 11) - perform experiment

Bacteriological Analysis of Urine (Exercise 12) - dilute samples and inoculate plates

April 13 and 17

Transmission of Microbes (Exercise 11) - examine plates, analyze and discuss results

Bacteriological Analysis of Urine (Exercise 12) - count plates, inoculate Enterotubes and inoculate antibiotic sensitivity plates

Bacteriological Analysis of Food (Exercise 13) - dilute samples and inoculate plates

April 20 and 24

Bacteriological Analysis of Urine (Exercise 12) - analyze and discuss results

Bacteriological Analysis of Food (Exercise 13) - analyze and discuss results

April 29 (Wednesday evening)

Examination 2 (35 points ... Exercises 7-13)

Notebook Assessment 2 (15 points)

Report 2 Due (15 points ... Bacteriological Analysis of Urine - Exercise 12)

• Graduate Teaching Assistants (GTAs)
  • Jenna Dolhi
    • Section A (Monday 9:00-10:50 am)
    • Office: 84 Pearson Hall
    • Phone: 529-7263 or 529-5422
    • E-mail: dolhijm@muohio.edu
  • Elizabeth French
    • Section B (Monday 3:00-4:50 pm)
    • Office: 82 Pearson Hall
    • Phone: 529-7266 or 529-5422
    • E-mail: frenche2@muohio.edu
  • Brock Arivett
    • Section B (Monday 3:00-4:50 pm)
    • Office: 50 Pearson Hall
    • Phone: 529-7263 or 529-5439
    • E-mail: arivetba@muohio.edu
  • Anand Prakash
    • Section C (Friday 3:00-4:50 pm)
    • Office: 82 Pearson Hall
    • Phone: 529-7266 or 529-5442
    • E-mail: prakasa@muohio.edu
  • Gloria Wada
    • Section D (Friday 9:00-10:50 am)
    • Office: 64 Pearson Hall
    • Phone: 529-7266 or 529-5422
    • E-mail: wadaga@muohio.edu

Laboratory Preparation

For best performance of laboratory exercises and best understanding of the material, you must approach the exercises in an organized fashion. This includes reading each exercise and writing a lab prep paragraph on what you are going to do during class before you come to the laboratory. This paragraph must include background material as well as a statement of the objective or purpose of the exercise, and it must be turned in to your GTA as you enter the laboratory. This paragraph may also be used as the Introduction section of your notebook entry for that exercise.

In addition, you will have five unannounced quizzes, which are intended to help provide incentive for you to prepare for lab each day.

You will be required to generate two formal laboratory reports during the semester. The exercises on which they will be based are listed on the laboratory outline. Each report should have a name, be well integrated and follow this format:

• Introduction
  • Background: What is the scientific basis of the approach or technique used--i.e., How does it work?
  • Purpose: What will/did the experiment(s) accomplish?
• Materials and Methods
  • Materials: What essential supplies were used?
  • Methods: How (exactly, in a stepwise manner) did you perform the experiment?
• Results
  • Tables/Graphs: What data were obtained? (Present the data in the most appropriate format for later interpretation.)
  • Text: What results are indicated by the data?
• Discussion
  • Interpretation of results: What do your results mean, in the context of both the current experiment and the field of microbiology in general?
  • Expectations: Did you get the results you expected to? Why or why not?
• Literature Cited
  • Cite the complete references of all the sources you used in preparing your lab report

You may use appropriate material from the textbook, laboratory manual, Exercises and lecture notes when preparing your reports, provided that you cite it appropriately. You should not, however, simply "regurgitate" any of this material (especially the procedural details), because the object of these reports is for you to integrate the information and present it in a clear and thoughtful manner in your own words. These reports must be generated using a computer-based word processor and inkjet or laser printer. The Department of Microbiology has a computer laboratory set up for student use. Ask your laboratory instructors when you can use this facility to complete your reports.

The laboratory portion of your course grade (25%) will be determined by your performance on two laboratory examinations (30 points for the first and 35 points for the second), two laboratory notebook assessments (5 points for the first and 15 points for the second), two 10-point laboratory reports, and five unannounced quizzes (10 points total). Although you are expected to participate in every laboratory exercise, you will be allowed one excused absence. Each additional absence will result in deduction of 10 points from your cumulative course points.

Your course grade will be determined using a plus/minus grading scale, as indicated on the evaluation web page.

Plagiarism and other infractions of Miami University rules and regulations will be dealt with as described in the Miami University Handbook (pay particular attention to Part V Academic Misconduct and especially to SECTION 01.502).

Cheating on any aspect of this course may make you subject to immediate dismissal from the class with a grade of F.

Laboratory Safety

Safety is of paramount importance in the microbiology laboratory.

Many of the microbes with which you will be working are capable of infecting humans under certain circumstances. In addition, you will be using disinfectants, carcinogens, caustics and flames during this course, all of which may be hazardous to your health. The instructions for each exercise in which it is necessary to work with either a caustic or a carcinogenic compound will warn you of the hazard and explain how to avoid harm while using that particular compound. You will not be warned repetitively about other hazards. Instead, you will be expected to abide by the following safety rules.

While you are in this laboratory ...

• Do not eat or drink or mouth pipette ... indeed, do not put anything in your mouth; mechanical pipetting devices will be provided for your use.
• Wear your laboratory coat at all times when you are in this laboratory, but do not wear it anywhere else.
• Wear goggles when using hazardous liquids.
• Wear disposable plastic gloves when using hazardous liquids.
• Use an absorbent pad soaked with disinfectant to cover your work area when working with infectious agents.
• Dispose of all hazardous materials in the receptacles provided.
• Use your burner flame cautiously ... and turn it off when you will not be using it for a long period of time, especially when you will be working with your microscope.
• Notify your instructor of any emergency situation as soon as it arises.
• Sponge off your laboratory bench with disinfectant at the finish of any exercise in which you have used microbes.
• Before leaving the laboratory ... remove your laboratory coat and hang it on the coat rack in the laboratory, then thoroughly wash your hands using the microbicidal detergent solution provided.