Lab Report 3

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University of Bridgeport *

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106

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Biology

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Dec 6, 2023

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1 Rose Shalani Rose BIOL-106-17 11/17/23 Lab 7: Antimicrobial Susceptibility testing Lab 8: Aerotolerance Lab 9: Biochemical Characterization Part 1 Lab 10: Biochemical Characterization Part 2

2 Rose Purpose: Lab 7: The purpose of this lab is for students to learn the origins of antibiotics and how resistance and sensitivity are tested. Students learned how to differentiate bactericidal and bacteriostatic antibiotics in this lab. Students also did a Kirby-Bauer experiment and made clear the results. Lab 8: In this lab, students learned how various microbes react to oxygen. Students performed an aerotolerance experiment making them understand how aerotolerance is classified. Lab 9: The purpose of the lab was for students to understand the basics of microbial identification. In the lab, students learned, performed, and interpreted catalase tests. During the lab, students could describe the biochemical purpose based on microbial organisms. Lab 10: In this lab, students could identify bacteria by knowing combinations of biochemical tests. In the lab, students could understand the purpose of an IMViC test and perform and examine the results of an IMViC. Introduction: Lab 7: In this lab, students performed a Kirby-Bauer test. A Kirby test is a type of disk diffusion test. A disk diffusion test is used to test the sensitivity of a microbe toward antibiotics. Antibiotics are a chemotherapeutic type used to stop the growth of bacteriostatic or kill bactericidal. Lab 8: In this lab, students learned about aerotolerance. Aerotolerance is the ability or inability of bacteria to grow with oxygen. Microorganisms are classified by their aerotolerance. There are 5 different microbial aerotolerant growth patterns. Obligate aerobes require oxygen for growth. Obligate anaerobes require no oxygen for growth. Facultative anaerobes grow in either oxygen or no oxygen but prefer to grow in oxygen. Aerotolerant anaerobes do not use oxygen to grow. Microaerophiles grow and survive in low oxygen. Lab 9: In this lab, students performed a Catalase and Oxidase test. A catalase test is used to detect catalase enzymes in bacteria. When a catalase test is positive a group of bubbles will be present. An oxidase test is used to see the presence of cytochrome c oxidase. When an oxidase test is positive there is a specific color that the result will turn. Lab 10: In this lab, students completed three different tests. Students completed an MR-VP test, an Indole test, and a Citrate Utilization Test. MR-VP stands for Methyl Red and Voges-Proskauer Test, which is a combination of the medium used for both tests. The MR part of the test is used to identify organisms that are capable of doing mixed acid fermentation. The VP part of the test is used to find organisms that ferment glucose. An indole test Is used to test if a bacteria can produce indole. A Citrate Utilization Test is used to determine if a bacteria can utilize citrate as a carbon source. Material: Lab 7:  Four Muller-Hinton Plates

3 Rose  Sterile Cotton Swabs  Antibiotic Stamp o Stamp 1  Ampicillin 10, Chloramphenicol 30, Erythromycin 15, and Kanamycin o Stamp 2  Penicillin 10, Streptomycin 10, Tetracycline 30, and Vancomycin 30.  E. Coli TSB culture  S. epidermidis Lab 8:  Fluid Thioglycolate Medium tubes  2 TSA plates  Anaerobic jar and GasPak  Clostridium sporogeneses culture  Alcaligenes faecalis culture  S. epidermidis culture  Bunsen burner  Inoculating loop Lab 9:  E. coli TSA slant o Grown overnight.  Enterococcus faecalis TSA slant o Grown overnight.  Alcaligenes Faecalis TSA slant o Grown overnight.  3% Hydrogen peroxide solution  Glass slide  Bunsen burner  Inoculating loop  Sterile cotton swab  Oxidase reagent Lab 10:  Three MR-VP broths  Three Deep Gelatin Tubes  Three Simmons Citrate Slants  VP reagents A and B  Kovacs Reagent  Six empty test tubes with caps  Three plastic 1 mL transfer pipettes  E. coli TSA slant o Grown overnight.  E. aerogenes TSA slant o Grown overnight.  Bunsen burner

4 Rose  Inoculating loop Procedures: Lab 7: 1. Label each MH plate with a different bacteria ( e. coli 1 and 2, s. epidermidis 1 and 2), initials, and date. 2. Use a new sterilized cotton swab and spread the bacteria all over the plate. 3. Use the antibiotic disk dispenser 1 for E. coli 1 and S. epidermis 1. 4. Use the antibiotic disk dispenser 2 for E. coli 2 and S. epidermis 2. 5. Incubate plates. Lab 8: Part A: 1. Label tubes with initials, date, and the correct name of the bacteria. 2. Using a sterilized inoculating loop, inoculate three broths with the different bacteria. Part B: 1. Label both plates with initials and date. Draw 3 quadrants on the plate and label each quad with each bacteria's name for both plates. 2. Using a sterilized loop inoculate each quad with the correct bacteria. 3. Place the plates in the correct spots (anaerobic and aerobic). Lab 9: Part A: 1. Transfer bacteria growth to a glass slide using a sterilized inoculating loop. 2. Add one or two drops of hydrogen peroxide to the glass slide. Cover with half of a Petri plate to minimize contamination. 3. Observed bubbles. 4. Repeat for the second bacteria. Part B: 1. Using a sterilized cotton swab apply oxidase. 2. Wipe the swab on the sample's surface to pick up bacterial growth. 3. Observe to see if there is any color change. 4. Repeat for the remaining organisms. Lab 10: Part A: o Day 1: 1. Inoculate 2 MR-VP cultures, one with E. coli and the next with E. aerogenes, then leave one test tube as a control. o Day 2 2. Mix each test tube well. 3. Put 1 ml of each culture into 2 test tubes. Be careful only to use one pipette per culture. 4. In the VP tubes add 15 drops of the VP reactant, then 5 drops of VP reagent b, and mix well after each reagent.

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