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Feb 20, 2024

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BACTERIAL GROWTH A N D I N HIBITIO N – POST LAB Procedures adapted from: Microbiology Experiments, 4 th edition. J. Kleyn and M. Bicknell. McGraw-Hill, 2004. Microbiological Applications , 7 th edition. H.J. Benson. McGraw-Hill, 1998. PART 1 A N D 2 Examine your plate. DO N OT OPE N THE PLATES. It is possible that some environmental samples contained pathogens (disease-causing microbes) which are now present in very high numbers. Also, mold spores can escape from open Petri dishes and contaminate an entire lab. When observing your plate, you should see mounds of microorganism cells called colonies growing on the surface of the medium. Some colonies are bacteria; they will generally have a soft appearance and can be dull or shiny. Although most bacterial colonies will be white or cream-colored, it is not unusual to see yellow or orange colonies. Colonies that are relatively large and filamentous (fuzzy) are typical of fungi. Record your observations of bacterial and fungal colonies in Table 1 (manipulate the table to fit your data). Include the following criteria: – Relative size (compared to other colonies) – Shape (round or irregular) – Color – Surface (shiny or dull) – Consistency (dry, moist, or mucoid) – Elevation (flat, craterlike, or conical) Count the total number of colonies in each half, and record your results in Table 2. Table 1 - I N CLUDE PICTURES OF YOUR PLATE HERE: Independent Variable Qualitative Colony Descriptions Finger Print 25 colonies; round and irregular, variation of orange and yellow in color, shiny surface and opaque as you cannot see through them. Elevation majority umbonate Finger Print after antibacterial wipe 2 colonies; shape is round, one yellow; shiny surface; opaque; elevation is convex

Table 2 - . QUESTIO N S 1. What was your null hypotheses? a. There will be no difference in finger print bacteria when comparing regular finger print vs. after sanitizing wipe. 2. What was your alternate hypothesis? a. There will be fewer bacteria from the finger print after using an antibacterial wipe. 3. Which one of your hypotheses was tentatively supported and which was refuted (or not supported)? a. The null hypothesis was refuted while the alternate hypothesis was supported. 4. In your own words, did your cleaning method work or not work? Why do you think this is? Independent Variable Qualitative Colony Descriptions Phone Approx. 22 colonies; size is small compared to other variable. Round in shape, surface is shiny; opaque; flat elevation Shoe Approx. 250 colonies; much larger colony than previous variable, variety of opaque and translucent; dull; elevation is mostly flat

a. I would say the cleaning method did make a difference. Taking it from 25 colonies down to 2 colonies. The antibacterial agent in the wipe must have been effective on the bacteria that was present on my hands. 5. Comparing the different experiments conducted in the lab, which type of environment had the greatest number of bacterial colonies? Why do you think this is? a. The greatest bacteria growth came from the bottom of my shoe. I would say this is because my shoe is always touching surfaces commonly traveled by others, and so bacteria is very present on the floors and picked up as I walk. PART 3 Follow your instructor’s directions for obtaining access to your cultures. Do not open the Petri dishes. Measure the diameter of the clear area surrounding the drug disk in millimeters (mm). This clear area is called the zone of inhibition . Record your data in the table below. If a disk does not have a clear area surrounding it, the diameter of the zone is recorded as zero. Use the drug chart to determine the name of the drug you used (if you don’t already know it) and record it in the corresponding table. Use the drug chart to determine whether Escherichia coli is resistant or susceptible to the drug you tested. Record the expected result and your actual result in the table below. Obtain results from your classmates to fill in the rest of the table. Examine the zone of inhibition again very carefully. Note whether you see any colonies (individual mounds of cells) growing inside the zone of inhibition. If so, this means the cells in

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the colony are resistant to the drug. Dispose of plates in the designated container. Wash your hands thoroughly with soap and warm water before leaving the lab. Antibiotic Test Results I N CLUDE PICTURES OF YOUR PLATE HERE: B. Cereus E.coli QUESTIONS: 1. Did you notice any resistant colonies? If so, for which drugs? a. B Cereus was resistant to Ampicillin Drug name Diameter of zone of inhibition for B. Cereus Diameter of zone of inhibition for E. coli Ampicillin 0 mm 2 mm Streptomycin 2 mm 3 mm Ciprofloxacin 3 mm 4 mm

2. What do you think would happen if you took resistant colonies and subcultured them (transferred them to a fresh agar plate) and exposed them to the same drug again? a. I would imagine it would not affect the bacterias growth. 3. What questions/variables would you like to test in the future? a. If I could go back I would swab my keyboard, or surfaces from my car. I would like to see how much bacteria is present in these places and how they differ.

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