BIOL 3053 Lab 7 Assignment (Winter 2022)

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Memorial University of Newfoundland *

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Memorial University of Newfoundland Department of Biology Biology 3053 Lab 7 Assignment ANTIBIOTIC SUSCEPTIBILITY TESTING AND API 20E MULTI-TEST Winter 2022 Name: Julie Hollahan Student #: 201311107 General Instructions: Type your name and student number in the above spaces. This assignment is due March 21, 2022 and is worth 2% of your lab grade . Read each question carefully. Type your answers in the space provided. When completed save your document as a pdf file and upload it into the Lab 7 Assignment Folder under Assessments – Assignments in the Brightspace shell. Once uploaded no changes to the assignment can be made. If you experience technical difficulties, contact the client Support Team of the Centre for Innovation in Teaching and Learning at www.citl.mun.ca/support Notice from Memorial University: You agree to the following; “All members of the Memorial University of Newfoundland community, including students, faculty, and staff, shall treat others with respect and fairness, be responsible and honest, and uphold the highest standards of academic integrity. By submitting this assignment, I unequivocally state that all work is entirely my own and does not violate Memorial University's Academic Integrity policy.” 1

OTHER INSTRUCTIONS : 1. As you go through the lab exercises, have your textbook and the appendices required for this lab available as these sources may help you to do the written assignment. 2. DO NOT copy and paste information from the lab or other material when answering your questions! This will be considered plagiarism and you will receive a grade of zero with possible further disciplinary action as per the University Calendar. Type your assignment in your own words ! 3. Read each question carefully and put your answers in the space provided. The spaces assigned for each question was just arbitrarily set to divide up the questions in the worksheet. You may use more or less of that space. 4. SAVE your document frequently. Once completed, save your assignment as a PDF document and upload your assignment to the Assignment Folder entitled “ Lab 7 Assignment ” in the pull down menu under Assessments. 5. If you have any questions, please contact your lab instructor through Brightspace OR through email: Sylvia at sbartle@mun.ca ; Kate at katecarson@mun.ca or Elizabeth at ediegor@mun.ca . 2

EXERCISE 14: ANTIBIOTIC SUSCEPTIBILITY TESTING 1. Fill up the table with your Results and Observations: Antimicrobial Agent Disk Code S. epidermidis E. coli Zone size (mm) S, I or R* Zone size (mm) S, I, or R Ampicillin Penicillin Tetracycline Streptomycin * S= sensitive; I = intermediate; R = resistant. For the purposes of this Exercise, we will consider antibiotics that affect both species (i.e. demonstrate a sensitivity) to be broad spectrum. 2. What is a broad spectrum antibiotic? (0.25 mark) - an antibiotic that is effective on gram-positive and gram-negative bacteria. 3. From your results in the table above , which of the antibiotic(s) you have used has/have a broad spectrum activity? Which do not? (1 mark) Broad Spectrum Not Broad Spectrum 3

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4. What is the mode of action and the range of the different antibiotics used in this exercise (see textbook, pp. 561-570)? (2 marks) For mechanism or mode of action: Be specific must explain exactly how this is accomplished e.g. don’t just say “inhibits protein synthesis” but give details. For range of antimicrobial: Type of organism affected by these antibiotics e.g. Chloramphenicol – broad spectrum; effective for Gram positive and Gram negative bacteria. a. Ampicillin b. Penicillin c. Tetracycline d. Streptomycin 5. Based on your results, which organism ( S. epidermidis or E. coli ) was more susceptible to Penicillin? Why? (1 mark) 6. What are the requirements of an antibiotic that would make it suitable for medical care? (1 mark) 7. When a bacterium is isolated from a patient’s specimen, it will be identified and its antibiotic sensitivity will be determined. If the bacterial specimen’s identity is known, why must its sensitivity also be determined? (1 mark) 4

EXERCISE 15: API 20E MULTITEST 1. Fill up the API # and the identity of the unknown. (1 marks) API 20E PROFILE SHEET RESULTS: UNKNOWN #: ______________________ Fill API#: Identity of Unknown: . 2. In the initial exercises of this course we examined the bacterial cells for size, shape, arrangement and then reactions to various stains. What aspect of the cells does that API Index examine to identify the various bacterial species? (1 mark) 3. What effect would a contaminated culture have on the ability of the API Index to identify a patient’s bacterial disease? (0.5 mark) 4. The bacterial cells were vortexed in the saline solution. Why was this done? (0.5 mark) 5. What enzyme was tested for by adding hydrogen peroxide to the Mannitol (MAN), Inositol (INO) and Sorbitol (SOR) tubules? What is the significance of this enzyme to the bacteria possessing it? (0.5 mark) 6. Why is sterile mineral oil added to some of the compartments of the API strip after inoculation with the test organism? (0.25 mark) 5

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