Introduction to Microbiology - Biology 6
Laboratory Syllabus
Denise Lim, Instructor
Spring, 2016
Required Materials:
• Lab Manual: Leboffe, M.J. & B.E. Pierce, 2010. Microbiology, Laboratory Theory and
Application, 4th ed., Customized for Cabrillo College, Morton Publishing Company. The 3rd
edition can be used as well.
• Weekly Lab Notes: These notes supplement the lab manual and include changes to protocols, a
list of the organisms we will be using, and any special notes or warnings you need to be aware of
before conducting the exercise. You must download these notes from lab website at
http://www.cabrillo.edu/~dlim/bio6/bio6labs.html.
Before coming to class each week, read the lab notes, the Introduction to the lab exercises from the
Lab Manual and additional readings from the textbook Microbiology, An Introduction, 12th ed.
by Tortora, Funke, & Case. Lab exams will include information from the reading
assignments.
Laboratory Policies:
1.
Equipment – required by Thursday of the first week. The Cabrillo Bookstore conveniently
carries all of these items, but you may be able get them for a better price at other stores.
•
Padlock for the storage drawer you will be sharing with your lab partner. Be sure to get a
lock small enough to fit the hole of the hasp.
Box of disposable latex or nitrile gloves; can be purchased at most drug stores,
supermarkets, or Costco.
Lab coat; can be purchased at California Coast Uniforms in Capitola. Your lab coat should
be long enough to cover and protect your lap and arms.
Safety glasses or goggles; Safety glasses or goggles from Chemistry lab can be used, or
they can be purchased at any hardware store (eg. Orchard Hardware & Supply on 41st
Ave.).
Lab marker like a Sharpie pen
Box of microscope slides. The microscope slides can be purchased at the Cabrillo
bookstore or online at Amazon (do a search for "microscope slides"). Cover slips are
unnecessary and we will not be using them. One box shared between four lab partners will
usually get you through the semester. You will need to have microscope slides by Week 4,
February 18, 2016.
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2.
Attendance: The first week of lab meetings are mandatory because they cover basic skills that
are the foundation for the rest of the semester. Inability to attend any of these labs will result in
being dropped from the course. Students need to be on time, as there will be days when you will
barely have enough time to finish. If you are late and miss the introductory lecture, you will not
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Bio6, Microbiology Laboratory
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be allowed to participate in that day's exercise. Because of the nature of the laboratory
exercises, the labs cannot be made up at another time.
3.
Be prepared for class. Before coming to class, always read through the lab introduction
provided in the lab manual, the experiments themselves, and the Lab Notes (download them
from the Bio 6 website). Do the background reading and WRITE UP A TO-DO LIST for the
day's experiments. This will help you avoid wasting valuable lab time orienting yourself. It is
also important for safety reasons: it will forewarn you of any special lab hazards and will help
eliminate rushing which is the primary reason for most avoidable lab accidents. Write out your
own bulleted list of tasks to be completed before each class meeting. Check off each item on
this list as you complete each part of the exercise in lab.
Occasionally there may be last minute changes from the protocol described in the Lab Notes
and lab manual. These changes will always be posted on the board at the beginning of class
during the lab introduction.
4.
Anyone caught tampering with or removing the cultures of other students will be dropped from
the course. This includes any cultures in either the incubators or the refrigerators.
5.
Grading. Your laboratory grade contributes 35% towards your final course grade.
a. Two Lab Exams: The lab exams will consist of half written questions and half practical
questions. The written questions will be comprised of a mixture of multiple choice, fill-in,
and short answer essay questions. The practical questions will based on your observation of
bacterial specimens. You will not be required to demonstrate any laboratory techniques.
You can prepare for these questions by taking good notes on the results for each lab
exercise and answering the questions in the Lab Study Guide.
b. Culture Identification Report: Each student will receive a pure culture of a bacterium to
identify. Using the techniques you have learned during the semester you will identify the
organism.
c. Lab Final Exam: The lab final will be a comprehensive practical exam based on the
experiments done in class throughout the semester. The final will consist of 50 practical
questions requiring identification of various specimens and interpretation of data that is
presented to you. Questions about any experiment that was performed during the semester
may be on the exam. Therefore, it is important that you keep good notes on your lab
observations.
LAB GRADE BREAKDOWN
Points
Lab Exam #1
50
Lab Exam #2
50
Lab Final (comprehensive)
50
Culture ID Report
50
Total
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Bio6, Microbiology Laboratory
200
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6.
How to Study for the Lab Exams
a. Use the Lab Study Guide downloaded from the BIO 6 Lab webpage
(http://www.cabrillo.edu/~dlim/bio6/bio6labs.html)
b. Review biological concepts, especially all biochemical reactions
c. Review lab protocols – understand the purpose of each step in a procedure; understand how
and when a particular protocol should be used
d. Review results – know what the cultures and media looked like both before and after
inoculation
7.
Accommodations. All students needing accommodations should inform the instructor as soon
as possible. Veterans may qualify for accommodations. Wounded Warriors may have acquired
injuries which through the American with Disabilities Act (ADA) entitles the use of
accommodations to ensure equal access for students with verified disabilities. To determine if
you qualify or need assistance with an accommodation, please contact ACCESSIBILITY
SUPPORT CENTER (Formerly DSPS), Room 1073, (831) 479-6379.
Laboratory Safety Rules
The microorganisms used in this course are of little or no risk to healthy people working with them,
provided that standard microbiological practices are followed. In order to maintain safe conditions in
the laboratory, the following rules must be observed.
1. No drinking or eating at any time. Period. Absolutely NO use of cell phones or hand-held
electronic devices (iPads or tablets) while gloved.
2. Clothing and Hair. A lab coat, gloves, goggles, and shoes that protect the tops of your feet
MUST be worn whenever you are working with cultures. You will not be allowed to participate
in lab activities if you are not properly attired. Hair must be tied back or secured so that it does
not hang in your face. Hair hanging loose is a contamination hazard and can catch fire in the
Bunsen burner flame. Securing your hair will also keep you from needing to touch it while
gloved.
3. Hand Washing. Gloves must be removed and hands must be washed with soap EACH TIME
you leave the lab for any reason (especially before using the restroom). When removing your
gloves, pull them off at the wrist so that they turn inside out and dispose of them in the biohazard
waste. Gloves should never to reused.
4. Work Area. Disinfect your bench top at the beginning and end of each laboratory period with
the disinfectant provided. Dispose of the paper towels used for cleaning bench tops in the
garbage can. When setting up your Bunsen burner, tape down the rubber hose to the table top to
prevent the hose from tangling with other equipment or tipping the burner over. Use the Bunsen
burners carefully and avoid reaching over the flame. Loop any electrical cords (especially for the
microscopes) once around the gas outlet to prevent them from being pulled off the table.
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Bio6, Microbiology Laboratory
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5. Culture Handling. You must assume that all the cultures you work with are potentially
pathogenic and handle them appropriately. Culture tubes and petri dishes are to be marked on the
bottom with your name or initials, experiment number and date. All cultures are always carried
in a rack or a basket. Culture tubes must be kept in a rack at the bench. Never prop up cultures
along the edge of the countertop or the black stain box. Cultures stored in the incubator or the
refrigerator must be in mesh baskets or racks. Place your cultures only on those shelves marked
for your lab section.
6. Culture Disposal. Unless you are told otherwise, old cultures are to be disposed of in the small
red biohazard bags and placed in the large "Biohazard" container. Be sure to place your bag in
the large liner bag as all biohazard waste must be double-bagged.
Used microscope slides must be disposed of in the red Biohazard Sharps container located by
the main sink. DO NOT dispose of slides in the red biohazard bags.
No cultures should be left in the 37º C incubators. Grown cultures may the TEMPORARILY
stored in the refrigerator if not all lab partners have observed the results, but be sure to throw
them out by the next class meeting. Check regularly to remove old cultures.
Cultures, pipettes, slides, media, and any other lab materials should NEVER to be discarded
in garbage cans, even if they are not contaminated. Cultures are NOT to be removed from the
laboratory under any circumstances.
7. Spills and Safety Equipment. Notify the instructor immediately if there is a culture spill, glass
broken, a fire, or if any injuries occur. Note the location of the eye wash, fire alarm, fire
extinguisher and the emergency shower. Make a note of the recommended evacuation route on
the safety cards posted by the door.
Spilled cultures should be flooded with disinfectant and covered with a paper towel. Leave the
disinfectant on the spill for at least 15 minutes before wiping up. Dispose of the paper towels in
your biohazard bag, NOT in the garbage can. Immediately change your gloves before touching
anything else.
8. Visitors. None.
9.
Personal Care. Be sure to notify the instructor if you are or become pregnant, have allergies,
are taking immunosuppressive drugs, have an immune system disorder, are diabetic, or have any
other medical condition which might necessitate special precautions in the lab.
10. Expectations. The knowledge and application of good aseptic technique is essential and will be
expected. Always remember that following these rules will lessen the hazards of working with
microorganisms not only to yourself, but to those around you. Keep in mind that the safety and
health of your fellow students rests in large part on the expectation that you will follow these
rules and apply them in the lab.
FAILURE TO OBSERVE THESE SAFETY RULES WILL RESULT IN IMMEDIATE
DISMISSAL FROM THE CLASS.
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Bio6, Microbiology Laboratory
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Biology 6 Schedule – Spring 2015
Wk
1
Date
Jan 26
Jan 28
2
Feb 2
3
Feb 4
Feb 9
4
Feb 11
Feb 16
Feb 18
5
Feb 23
6
Feb 25
Mar 1
7
Mar 3
Mar 8
8
Mar 10
Mar 15
Mar 17
Spring , 2016
Laboratory Exercises
Introduction to the Lab
Ex. 2-1: Ubiquity of Microorganisms
Ex. 1-3: Aseptic Transfer and Inoculation
Ex. 1-4: Streak Plate Method of Isolation
Ex. 2-2: Colony Morphology
Ex. 2-3: Growth Patterns on Slants
Ex. 2-4: Growth Patterns in Broth
Observe Tuesday's Cultures
Ex. 2-7: Fluid Thioglycollate Medium
Ex. 7-3: Antimicrobial Susceptibility
*Disinfectants
Observe Tuesday's Cultures
*Ex. 2-9: Temperature
Ex. 2-11: Osmotic Pressure
Observe Tuesday's Cultures
Ex. 3-5: Smear preparation
BRING MICROSCOPE SLIDES
Ex. 3-1: Introduction to the Microscope
Ex. 3-5: Simple Stains
Ex. 3-7: Gram Staining
Ex. 3-8: Acid Fast Staining
Ex. 3-10: Endospore Staining
Lab Exam #1 (Weeks 1-5)
Ex. 4-4: Mannitol Salts Agar
Ex. 4-5: MacConkey Agar
Ex. 5-3: Phenol Red Broth
Observe Tuesday's Cultures
Ex. 5-6: Catalase Test
Ex. 5-7: Oxidase Test
Ex. 5-15: Lipid Hydrolysis test
Ex. 5-16: Casein Hydrolysis Test
Ex. 5-17: Gelatin Hydrolysis Test
Observe Tuesday's Cultures
Ex. 5-4: Methyl Red and Voges-Proskauer Tests
Ex. 5-8: Nitrate Reduction Test
Bio6, Microbiology Laboratory
Reading
Text Ch. 6
Text pg. 568;
*See Lab Notes for
protocol
*See Lab Notes for
protocol
Text Ch. 3
page 5
Wk
9
Date
Mar 22
Mar 24
Mar 29 & 31
10 April 5
11
April 7
Lab Exam #2 (Weeks 6-9)
Apr 12
Observe Last Tuesday's Cultures
Streptoccocus ID:
Ex. 4-3: Bile Esculin Test
*Ex. 5-24: Bacitracin Test
Ex. 5-25: Blood Agar
†NaCl Broth
Observe Tuesday's Cultures
Dichotomous Keys
*Culture Identification Exercise
Rough draft of Dichotomous Key due
Culture Identification Exercise
Culture Identification Exercise
Apr 14
12
Apr 19
13
Apr 21
Apr 26
Apr 28
14
Laboratory Exercises
Ex. 5-9: Citrate Test
Ex. 5-18: Urea Hydrolysis Test
Ex. 5-20: SIM Medium
Observe Tuesday's Cultures
SPRING BREAK
Staphylococcus ID:
Ex. 4-4: Mannitol Salts Agar
Ex. 5-14: DNA Hydrolysis Test
Ex. 5-27: Coagulase Test
*Ex. 5-24: Novobiocin Test
May 3
May 5
15
May 10
16
May 12
May 17
Spring , 2016
Ex. 5-30: EnteroPluri Tubes
Culture Identification Exercise
*Staphylococcus Quick Agglutination Test
*Streptococcus Quick ELISA Test
Culture Identification Exercise
Ex. 11-6: ELISA
Culture Identification Exercise
Review and Lab Cleanup
Culture Identification Reports Due
LAB FINAL EXAM (COMPREHENSIVE)
Bio6, Microbiology Laboratory
Reading
*Review the Kirby Bauer
Method: Ex. 7-3
*Review the Kirby Bauer
Method: Ex. 7-3
†See Lab Notes for
protocol
*See Lab Notes for
protocol
*See Lab Notes for
protocol
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