Attachment_ PDF_ Bacterial Safari Lab! 2023

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St. John's University *

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Biology

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Jan 9, 2024

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Uploaded by BaronSteelHippopotamus25

Join us as we go on our bacterial safari! We will be focussing on some of the most numerous and important organisms on Earth – bacteria. Bacteria live in mixed populations almost everywhere on earth, even within us! This means they should be easy to find. Although bacteria are often associated with disease, most bacteria contribute tremendously to human health and well being. For example, bacteria in the human intestine produce vitamin K, while bacteria in the environment play an important role in nutrient cycles. Introduction As on any trip, proper preparation is essential. Your safari leader (teacher) has dictated that you must understand the following terms before you can begin your safari. 1. What is the difference between an antibiotic, antiseptic, and disinfectant? [3 marks A] 2. In the past decade, the number of antibiotic resistant strains of bacteria has increased dramatically. How does bacterial resistance occur? [1 marks A] How are antibiotic resistant genes transferred from one bacterium to another? [1 marks A] 3. List two ways in which antibiotic resistance in bacteria can be prevented? [2 marks A] Problem: Part A: To examine the predominance of bacteria in our environment. Part B: To test the effectiveness of various agents against inhibiting bacterial growth. Hypothesis: Part 1: Where do you think you would find the highest level of bacteria in the school? Explain your reasoning using an IF & THEN statement. [2 mark T&I] Part 2: You will predict which Antiseptic/Antibiotic/Disinfectant will be most effective at limiting bacteria growth. (Options: Anti-Bacterial Soap, Household Bleach, Household Disinfectant, Amoxicillin, Erythromycin, Penicillin) State your Hypothesis using an IF & THEN statement & explain your reasoning. [2 marks T&I] Materials/Apparatus: ▪ Nutrient agar Petri dishes ▪ Sterile cotton swabs ▪ Marker ▪ Computer Safety Be careful when working with live bacterial cultures. Wash your hands with soap immediately after handling any bacterial culture. Once Petri dishes have been inoculated, replace the lids immediately. Do not take bacterial samples from coughing or from other bodily sources. Carefully clean and disinfect your work area after you compete the lab. Follow your teacher’s directions about disposal of swabs, cultures, and Petri dishes. Good luck on the safari!!! Part 1 Procedure: Bacteria Around You! 1. Turn the Petri dish upside down and using a marker, label each half of the dish. Add your group #, cohort and date at the edge to identify the dish. 2. Collect bacteria from 2 different locations in the classroom. Wet one sterile swab for each sample. Gently wipe the surface to be sampled (for example the floor, a door knob, a keyboard) and transfer your sample to your sterile Petri dish. 3. Slide the lid off the Petri dish and streak the agar using the swab. Be careful not to damage the agar. Replace the lid immediately after you have streaked the dish. Label the sample on the corresponding quadrant. 4. After streaking three quadrants, turn it upside down and return it to your teacher. The dishes will be placed in an incubator overnight. T&I: /23 marks App: /20 marks Name: __________________________

Part 2 Procedure: What Kills Germs?? *Note: Procedure below is for information purposes only. Please Note: video instructor does not measure the zones of inhibition correctly. Consult your teacher for guidance on this issue. Link: https://youtu.be/dgMi18WbHLk 1. Inoculate the agar in the petri dish by clicking one of the test tubes containing pathogenic bacterial stock culture- Staphylococcus aureus, Hemophilus influenzae, or Streptococcus pneumoniae. 2. Vials 1 through 7 contain filter paper disks that have been soaked in antimicrobial agents such as antibacterial soap, household bleach, household disinfectant, penicillin, amoxicillin, and erythromycin, or in sterile water (as a control). Drag a disk from each vial and place it in the petri dish. To avoid contamination, disks should not be moved after they have been dropped into the petri dish. 3. At any time in the Virtual Lab, click the Microbiology book to find out about specific pathogenic bacteria and antimicrobial agents. Click the fingers pointing left and right to page through the information. 4. Click the incubator to place the petri dish in it. 5. Click the red button on the incubator to turn it on. When the timer shows that 24 hours have passed, click the incubator to remove the petri dish. 6. Examine the patterns of bacterial growth. The colored area that covers most of the surface of the petri dish is the lawn culture of the bacteria-a visible layer of thousands of bacterial cells. 7. Drag the ruler to measure the diameters of the zones of inhibition around the disks (the tan areas). Some disks may be surrounded by large zones of inhibition, where no bacteria grew due to the strong inhibitory effect of the antibiotic, antiseptic, or disinfectant on the disks. Other disks may have caused little or no inhibition-meaning that the bacteria are partially or completely resistant to the antimicrobial agent on them. To find out which antimicrobial agent corresponds to a specific number, move the cursor over the number. In the Table, enter the measurement for each antimicrobial agent. 8. Click the Reset button and repeat the Virtual Lab until you have tested all the antimicrobial agents on all three types of pathogenic bacteria. 9. Use the data in the Table to compare the effectiveness of different antimicrobial agents on different bacteria. Complete the Journal questions. Observations Table 1: Bacteria Around You! [6 marks T&I] After incubation describe the following characteristics for the specified set of bacterial plates. Record the observations for an average bacterial colony or the most numerous one if several different colonies are present. Use the diagrams below to help you describe the form and margin of the predominant colonies that are present. Source # of Colonies Average Colony Size (mm) Colour Form (see below) Elevation (see below) Margin (see below) CONTROL Table 2: What Kills Germs? [9 marks T&I] Table 2: Zones of Inhibition for Various Agents [9 marks T&I]

Table 2: Zones of Inhibition for Different Anti-Microbials [9 marks T&I] Diameter of Zone of Inhibition (mm) Bacteria Species Sterile Filter Paper (Control) Anti- Bacterial Soap Household Bleach Household Disinfectant Amoxicillin Erythromycin Penicillin Hemophilius influenzae 0 mm Staphylcoccus aurenus 0 mm Streptococcus pneumoniae 0 mm Analysis: Part 1: 1. Which areas around the school appeared to have large populations of bacteria? Suggest reasons for these findings? [2 marks A] 2. What in the bacteria’s environment gave it the ability and energy to grow? [1 mark A] Part 2: 3. What anti-microbial was the most effective at controlling bacterial growth? How do you know? [2 marks A] 4. Compare the general effectiveness of the different antibiotic drugs (Penicillin, Amoxicillin, Erythromycin) and chemical disinfectants (Anti-bacterial soap, Household disinfectant, household bleach). Which seem to be better at controlling bacterial growth? Why do you think this is so? [2 marks A] Conclusion: Part A: restate purpose; accept/reject hypothesis; state 2 improvements [2 marks T&I] Part B: restate purpose; accept/reject hypothesis; state 2 improvements [2 marks T&I] Discussion: 1. The local hospital has been infected with a highly infectious form of Staphylococcus bacteria. The pathogen is spread through infected instruments and by direct contact. How would you use antiseptics and disinfectant to stop the spread of the pathogen to uninfected parts of the hospital? [2 marks A] 2. What future complications could arise from the overuse of antibacterial mouthwashes or cleaning agents? [2 marks A] 3. If you were a doctor treating a patient infected with Staphylococcus aureus, a bacterium that causes mild to moderate skin infections, which antibiotic would you prescribe? Why? [2 marks A]

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