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University of Florida
Microbiology and Cell Science
MCB 3020L

Chloe Evetts (8990-4674) Cameron Jacobs (7908-1325) Combating Microbes Purpose Being able to control and stop microbial growth is of utmost importance to the health and wellbeing of human beings and animals alike. Unknown disease causing microbes could wreak havoc if they are allowed to replicate interrupted with no possibility of combating these invaders. The purpose of this lab is to learn the different methods of antimicrobial control, including biological, physical, mechanical, and chemical methods and how to use these to control foreign invaders. The Kirby-Bauer Antibiotic Disk Diffusion will be used to determine the efficacy of the antibiotics Penicillin, Vancomycin, Oxacillin, Gentamicin, Tetracycline, Chloramphenicol and various reagents including mouthwash, disinfectants, and cleaning reagents against Escherichia coli and Staphylococcus aureus. Hypothesis We believe that by correctly using proper aseptic techniques, by properly inoculating Escherichia coli and Staphylococcus aureus onto agar plates, and by using the Kirby-Bauer Antibiotic Disk Diffusion method, we will be able to determine the efficacy of each agent against the two species. Efficacy will be able to be determined by analyzing the zone of clearing, a clear area around the disk that equates to no microbial growth. The larger the zone of clearing, the smaller the minimum inhibitory concentration. Procedure: 1 Lab 1a: a Label the bottom of each Mueller-Hinton agar plate with your initials, date, medium type and test microorganism. b Using aseptic technique, inoculate each plate with their respective test organisms as follows: i Immerse the swab into the culture tube and squeeze excess culture on the inner wall of the tube. ii Using the swab, streak the entire surface of the plate three times, (rotating the plate 60° each time). iii Finally, run the swab around the outer edge of the plate. iv Allow each plate to dry for 5 to 10 minutes at room temperature. c Apply the antibiotics individually to the surface with forceps dipped in alcohol and flamed. Make sure that contact is made with the surface. DO NOT PRESS THEANTIBIOTIC INTO THEAGAR. DO NOT REMOVE THEANTIBIOTIC ONCE IT HAS BEEN PLACED ON THEAGAR. d Incubate the Mueller-Hinton agar plates for 24 to 48 hours in an inv
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