Introduction to Microbiology - Biology 6 Laboratory Syllabus Denise Lim, Instructor Spring, 2018 Required Materials: Lab Manual: Leboffe, M.J. & B.E. Pierce, 2016. Microbiology, Laboratory Theory and Application, 4 th ed., Customized for Cabrillo College, Morton Publishing Company. The 3 rd edition can be used as well, but the exercises are numbered differently. Weekly Lab Notes: These notes supplement the lab manual and include changes to s, 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 and print these notes from Canvas. Learner Outcomes: 1. Apply the principles of modern laboratory technology such as aseptic technique, standard laboratory procedures used to identify a bacterial culture, conditions that affect the growth and control of microbial life forms, and molecular biology. 2. Analyze the impact of diverse microbial life forms on human activity, especially in the context of health and disease. 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, 12 th 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 borrowed from Cabrillo or 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 41 st 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 Spring, 2018 Bio6, Microbiology Laboratory page 1
usually get you through the semester. You will need to have microscope slides by Week 4, February 22, 2018. 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 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 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 be 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 construct a dichotomous key, identify the organism, and write a 2 page report on that 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. Spring, 2018 Bio6, Microbiology Laboratory page 2
LAB GRADE BREAKDOWN Points Lab Exam #1 50 Lab Exam #2 50 Lab Final (comprehensive) 50 Culture ID Report 50 Total 200 6. How to Prepare for the Lab Exams a. Use the Lab Study Guide downloaded from Canvas b. Review biological concepts, especially all biochemical reactions c. Review lab s understand the purpose of each step in a procedure; understand how and when a particular 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. Spring, 2018 Bio6, Microbiology Laboratory page 3
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. 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. Spring, 2018 Bio6, Microbiology Laboratory page 4
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. Spring, 2018 Bio6, Microbiology Laboratory page 5
Biology 6 Schedule Spring 2018 Wk Date Laboratory Exercises Reading 1 Jan 30 Introduction to the Lab Feb 1 Ex. 2-1: Ubiquity of Microorganisms Ex. 1-3: Aseptic Transfer and Inoculation Ex. 1-4: Streak Plate Method of Isolation 2 Feb 6 Ex. 2-2: Colony Morphology Ex. 2-3: Growth Patterns on Slants Feb 8 Ex. 2-4: Growth Patterns in Broth 3 Feb 13 Ex. 2-7: Fluid Thioglycollate Medium Feb 15 Ex. 7-3: Antimicrobial Susceptibility *Disinfectants 4 Feb 20 *Ex. 2-9: Temperature Feb 22 Ex. 2-11: Osmotic Pressure Ex. 3-5: Smear preparation BRING MICROSCOPE SLIDES 5 Feb 27 Ex. 3-1: Introduction to the Microscope Mar 1 Ex. 3-5: Simple Stains Ex. 3-7: Gram Staining 6 Mar 6 Ex. 3-8: Acid Fast Staining Ex. 3-10: Endospore Staining Mar 8 Lab Exam #1 (Weeks 1-5) 7 Mar 13 Ex. 4-4: Mannitol Salts Agar Mar 15 Ex. 4-5: MacConkey Agar Ex. 5-3: Phenol Red Broth 8 Mar 20 Ex. 5-6: Catalase Test Mar 22 Mar 27, 29 Ex. 5-7: Oxidase Test Ex. 5-15: Lipid Hydrolysis test Ex. 5-16: Casein Hydrolysis Test Ex. 5-17: Gelatin Hydrolysis Test Ex. 5-4: Methyl Red and Voges-Proskauer Tests Ex. 5-8: Nitrate Reduction Test SPRING BREAK Text Ch. 6 Text pg. 568; Text Ch. 3 Spring, 2018 Bio6, Microbiology Laboratory page 6
Wk Date Laboratory Exercises Reading 9 Apr 3 Ex. 5-9: Citrate Test Apr 5 Ex. 5-18: Urea Hydrolysis Test Ex. 5-20: SIM Medium 10 Apr 10 Streptoccocus ID: Ex. 4-3: Bile Esculin Test *Ex. 5-24: Bacitracin Test Ex. 5-25: Blood Agar NaCl Broth Apr 12 Lab Exam #2 (Weeks 6-9) 11 Apr 17 Observe Last Tuesday's Cultures Apr 19 Staphylococcus ID: Ex. 4-4: Mannitol Salts Agar Ex. 5-14: DNA Hydrolysis Test Ex. 5-27: Coagulase Test *Ex. 5-24: Novobiocin Test Dichotomous Keys 12 Apr 24 * Apr 26 Rough draft of Dichotomous Key due 13 May 1 May 3 14 May 8 *Staphylococcus Quick Agglutination Test May 10 *Streptococcus Quick ELISA Test Ex. 5-30: EnteroPluri Tubes 15 May 15 Review and Lab Cleanup May 17 Culture Identification Reports Due LAB FINAL EXAM (COMPREHENSIVE) 16 May 22 Lecture Final No labs this week *Review the Kirby Bauer Method: Ex. 7-3 See Lab Notes for *Review the Kirby Bauer Method: Ex. 7-3 Spring, 2018 Bio6, Microbiology Laboratory page 7