Integrated Science 2 TOC # Name. Antibiotics Effect on Bacteria

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1 Integrated Science TOC # Name Antibiotics Effect on Bacteria Per Antibiotics Effect on Bacteria Introduction: By definition, an antibiotic is a biochemical produced by a microorganism that inhibits the growth of or kills another microorganism. The use of antibiotics has revolutionized clinical medicine. Antibiotics allow modern doctors to treat a far wider range of diseases more effectively and economically. One way to test the effectiveness of an antibiotic against a specific microorganism is the Bauer-Kirby test. This test measures how well antibiotic disks (disks which contain a known amount of antibiotic) inhibit bacterial growth when placed on an agar dish swabbed with the desired microorganism. The antibiotic disks produce zones of inhibition (clear areas of no growth) which are measured in order to determine the susceptibility of a microorganism to the different antibiotics used in the test. Purpose: Microscopic bacteria are pervasive as both disease-causing agents and as helpful organisms. In this activity, you will be conducting two separate experiments on bacteria. Experiment : comparing bacterial populations from a number of different locations. Experiment : comparing the inhibitory capabilities of a variety of antibiotics on various bacteria; to learn which antibiotics will best slow bacterial growth. Hypothesis: Read through the procedure and write a hypothesis for each experiment: Experiment : Experiment : Materials: large petri dishes with lids labeled C,, or bacterial broth cultures, one Gram-negative and one Gram-positive Agar Permanent pen Q-tip with water circular antibiotic disks--each soaked in a different inhibitor: o Penicillin o Tetracycline o Chloramphenicol Forceps

2 Procedures: Day Today, you will prepare BOTH experiments. All three petri dishes need incubate before you can make observations. Preparing experiment :. Preparing the Petri Dish One of your Petri dishes will be labeled C for control. There will be three () bottles of antibiotics labeled either,, or in the front of the room. Important: Note the numbers of the bottles. These numbers correspond with the number on the petri dish Using a pen, carefully divide the outside of the bottom of the control petri dish into two sections, as shown, and number them and. (aka, do not write on the lid or in the agar). Inoculating the CONTROL Petri Dish Note: DO NOT collect samples from toilets. This is not only disgusting, it s highly unsanitary. Using a moistened Q-tip swab, collect a sample of bacteria from a site of your choice and touch the tip of the swap gently to the agar on section of the plate. Do not touch the swab with your fingers. Open the dish only long enough to inoculate the agar and then replace the cover. Using a moistened Q-tip swab, collect a second sample of bacteria from a different site of your choice and touch the tip of the swap gently to the agar on section of the plate. Do not touch the swab with your fingers. Open the dish only long enough to inoculate the agar and then replace the cover. Record the inoculation sources here and on your Data table:... Incubation Label your petri dish on the top with your first names and class period. Use a couple of pieces of tape to loosely attach the top of the dish to the bottom. Place the dish upside down in an area designated by your teacher. The dish is incubated upside down so that any liquid that collects will drip down onto the cover and not collect on the surface of the agar.

3 Preparing experiment :. Preparing the Petri Dishes and (day ) Use a permanent pen to divide each petri dish into three sections labeled,, and Now label the bottom of one dish with the source from which you collected sample for the control dish. Label the other petri dish with the source from which you collected sample for the control dish Using a moistened Q-tip swab, collect a sample of bacteria from the same site from which your collected sample for the control petri dish. Touch the tip of the swap gently to the agar on all three () sections of the petri dish labeled with the same collection source area. Do not touch the swab with your fingers. Open the dish only long enough to inoculate the agar and then replace the cover. Using a moistened Q-tip swab, collect a sample of bacteria from the same site from which your collected sample for the control petri dish. Touch the tip of the swap gently to the agar on all three () sections of the petri dish labeled with the same collection source area. Do not touch the swab with your fingers. Open the dish only long enough to inoculate the agar and then replace the cover.. Inoculating the Petri Dish and placing the Inhibitors Using the forceps, place one () antibiotic disk containing penicillin in section of the petri dish Repeat this procedure, placing one () disk containing tetracycline in section and one () disk containing Erythromycin in section of the petri dish. Penicillin Tetracycline Erythromycin. Incubation (same as Experiment ) Label your petri dish on the top with your first names and class period. Loosely secure the top to the bottom with a few pieces of tape and place the dish upside down in an area designated by your teacher. The dish is incubated upside down so that any liquid that collects will drip down onto the cover and not collect on the surface of the agar.

4 Day Retrieve all three of your lab group s petri dishes from the incubator. DO NOT open the dishes!! Live bacteria are growing in these dishes. You DO NOT want to breathe the bacteria into your body.. Observing Growth (day ) First, make observations of the CONTROL petri dish Measure (in cm) and record the diameter of the area of bacterial growth in both sections. Record data about your petri dish on your Data table by drawing the correct size of the bacterial colonies in the section provided. Draw the colonies the correct size and in the correct position, as you observed them on the petri dish. Note color, shape and thickness. DATA TABLE Amount of Growth (Small, Medium or Large).. 4. Observing Growth (Day ) Now, make observations of the two petri dishes with antibiotics Measure (in cm) and record the diameter of the area of bacterial growth in all three sections. Record data about your petri dish on your Data table by drawing the correct size of the bacterial colonies in the section provided. Draw the colonies the correct size and in the correct position, as you observed them on the petri dish. Note color, shape and thickness. Diameter of Inhibition Area (cm) Source of bacteria... 4

5 Source of bacteria Diameter of Inhibition Area (cm)... Discussion/ Conclusion: Experiment (Control):. Look at your petri dish. In which sections are the bacterial colonies the largest? Give a reason for your answer.. What is the importance of a control in a scientific experiment?. What does the nutrient agar provide for the bacteria? 4. Why didn't you seal shut the petri dish after inoculating the agar? 5. The petri plate was turned upside down during the incubation period so liquid would not accumulate on the surface of the agar. Why is it important that water not collect on the surface of the agar? 5

6 6. Was your hypothesis correct? 7. How could the information you gathered in this experiment be useful in "the real world"? Experiment : 8. For each petri dish, which antibiotic was the best inhibitor of bacteria growth? Explain. 9. Which antibiotic was the worst inhibitor of bacteria growth? Explain 0. Do the antibiotics kill the bacteria (bactericidal) or only inhibit growth (bacteriostatic)? Explain your answer.. If the antibiotic concentration is doubled, will the growth zone be twice as large? Why?. Was your hypothesis correct?. How could the information you gathered in this experiment be useful in "the real world"? 6

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