revised lab prac 2 study guide.docm

revised lab prac 2 study guide.docm - Lab Practical#2 Study...

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Lab Practical #2 Study Guide for Microbiology 146 As you study for the lab practical, read the labs over again, especially the introductions, terms in bold print, results/observations and discussion questions. Also, look at any notes or photographs you took and drawings you did during each lab. You will not be asked about specific procedural steps for any lab; instead, the emphasis will be on the introductory information and your observations and experimental results. Please note that lab coats are required during the lab exam. Metabolic Testing of Bacteria 1. For ALL Metabolic Tests, go back and study your charts 2. For each metabolic test, know: 1) the test name, 2) what you are specifically testing for, 3) the medium +/- any reagents used, 4) a positive vs. negative result and 5) the reason for a positive result. 3. Given experimental tubes +/- any accessory materials, identify the specific test and interpret test results. Blood Smear 1. Identify the different leukocytes under the microscope. Leukocytes as part of the innate immune system are non-specific Leukocytes are the Second line defenses which help combat foreign invaders when first line defenses have been breached Non-specific Leukocytes (Innate) include Neutrophils Eosinophils Basophiles Monocytes Natural killer (NK) cells Specific Leukocytes (adaptive) include Lymphocytes (T and B) 2. Know the frequency of each leukocyte in blood. The most numerous Lymphocytes:
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N ever L et M onkey’s E at B ananas Neutrophils (50-70%) Lymphocytes (30%) Monocytes (5%) Eosinophils (2-3%) Basophiles (1% Unknowns 1. Know the steps you performed, in general, to identify your unknown. Day 1 : Streak Plate (to find gram neg. or gram pos.; motility and individual colonies) Day 2 : Dichotomous Key, Colonies morphology, easy specimen result* Day 3 : Inoculated media and compared media to dicho. Key; performed metabolic tests Day 4 : results from metabolic tests and find bacteria, result for hard specimen* 2. Apply information learned to determine a species name given test results and a dichotomous key.
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Bacteriophage Titer 1. Given a series of plates, identify the bacteriophage titer experiment. When performing the counts, it must be between 30-300, if greater than 300 it’s TMTC. 2. Explain how bacteriophages work in this experiment. We mixed the E. Coli with coliphages in the tryptone agar base plate. In the lytic phage (lytic cycle) they replicate and lyse the cell . In the temperate phage (lysogenic cycle) they integrate into the host chromosome as prophages. 3. Explain how serial dilution is used to estimate the number of phage in the sample. Serial dilution of phages is used so that a countable number of plaques will be available on at least one of the tryptone agar plates. 4. Know which plates to select to count plaques, and how to back-calculate the number of phage from the plaque (PFU) count .
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