Abby Taylor per-3 Lab #3

Introduction: In 1982 Alexander Fleming, a Scottish scientist discovered the antibiotic penicillin. He observed a mold growing on a bacterial culture in his lab. He noticed a clear zone around the mold in which no bacteria was growing. He discovered that this mold produced a protective chemical which inhibited or killed bacteria. The mold was identified as penicillium notatum. Since that time many organisms have been identified that that produce similar compounds. These compounds are called antibiotics. These chemicals are harvested in lab and help to fight infectious diseases caused by bacteria. Due to the frequent use of antibiotics, it has been discovered that any bacteria are mutating and becoming antibiotic resistant.

MRSA- methicillin resistant Stahpylocccus aureus or hospital staph is a bacterial strain that had mutated and become antibotic resistant.
Objective: Indentify which antibiotics and antiseptics are most effective against Echerichi Coli and Aka E. Coli

Problem: Due to the frequent use of antibiotics, it has been discovered that many bacteria are mutating and becoming antibiotic resistant.
Scientific Theory: 1: Bacteria is a domain of unicellular prokaryotes that have a cell walls containing peptidoglycans 2: An antibiotic is a compound that blocks the growth and reproduction of bacteria. (tetracycline) Antiseptic is an antimicrobial substance which are appled to living tissues skin to decrease the likeliness of infection or putrefaction. 3. Antibiotics kill the bacteria growth because, they bind to specific sites in the bacteria cell wall to prevent the bacteria

making new cell walls. So the cell wall breaks down and the cell dies. Hypothesis: If tetracycline and alcohol work together, then, tetracycline will prevent protein growth and alchol prevents E-coli growth Materials: Markers Aluminum Foil 2 sterile cotton swabs 2 sterile paper disks Sterile forcepts beaker with alcohol metric ruler parafilm culture of E.Coli 2 sterile nutrient agar plates 3 antibiotic disks: erythromycin streptomycin, tetracycline 3 paper disks dipped in different antiseptics Safety: goggles, aprons, gloves,tie back long hair Procedure: 1. sterilize the working area by cleaning with alcohol or clorox. Do not place any personal materials on the desk 2. lay out a sheet of aluminum foil 3. without opening the petri dishes,mark the bottom of the dish. Use a metric ruler to divide the dish into 4 areas of equal size. Label the areas 1-4, put your period #, table number and date around the edges of the dish 4. go to the inoculation table where your teacher will help you to inoculate your dishes with the bacterial culture 5. dip the sterile cotton swab in bacterial culture, streak the agar as instructed. Be careful not to crack the agar 6. place the cotton swab into the beaker containing alcohol for diasposal. 7. repeat for the second dish

Antibiotics 1. sterilize a pair of forceps in the beaker of alcohol 2. take a blank disk and gently press onto the agar in area 1 of your plate. (control) 3. sterilize forceps and repeat with 3 antibiotic disks. Label which area has which antibiotic 4. sterilize forceps and seal the petri dish with parafilm 5. place dish upside down in the incubator for 48 hours Antiseptics 1. sterilize a pair of forceps in the beaker of alcohol 2. take a blank dish that has been dipped in water and gently press onto the agar in area 1 of your plate. (control) 3. sterilize forceps, take a blank disk and dip into an antiseptic and gently press onto the agar, repeat for 2 other antiseptic solutions. Label which area hs which antiseptic 4. sterilize forceps and seal the the petri dish with parafilm 5. place dish upside down in the incubator for 48 hours Clean Up 1. wrap all materials in the foil and dispose of foil in the garbage 2. clorox the lab table 3. remove and dispose of gloves and wash hands 4. put apron away Data Collection 1. observe the petri dish after 24-48 hours DONT OPEN THE PETRI DISH! 2. notice any clear areas called the zone of inhibition surrounding the paper disks. A clear area indicates that the antibiotic or antiseptic inhibited bacterial growth. If there is no zone of inhibition, then the antibiotic or antiseptic was not effective in inhibiting bacterial growth 3. with a metric ruler measure to the nearest millimeter the size of the clear zone surrounding each disk. Record the

measurements in the data table

Effects Of Antibiotic on Growth Of Antibiotic Disk 1. Tetracycline 2. Streptomycin 3. Erythromycin 4. Control (water)

Zone Of Inhibition (mm 13mm 10mm 6mm 0mm

Effects Of Antiseptic on Growth Of Antiseptic Disk 1. Control 2. Windex 3. Alcohol 4. Amonia

Zone of Inhibition (m 0mm 0mm 4mm 5mm

Use the formal lab report and be sure to include your data, analysis and conclusion Data: 1. Include a diagram of your plate with the zones of inhibition (mm) clearly identified and labeled for each antibiotic and antiseptic 2. Use your data to make a bar graph demonstrating the effectiveness of each antibiotic and antiseptic. Dont forget to title your graph and label the axis Analysis:

1. It is important to use sterile techniques while inoculating agar plates because, no extra bacteria gets on the ogar, so you are able to test the antibiotics and antiseptics against the bacteria you put on.

2. Antibiotic is a compound that breaks the growth and reproduction of bacteria. Antibiotics work against bacteria because, it kills the bacteria cell walls and makes it explode. When they try to make more cell walls, it becomes weaker. 3. The effectivness of an antibiotic and antiseptic can be measured by the distance from the bacteria to the edge of the ring. 4. The tetracycline was the best because, it had the largest distance from the bacteria to the edge of the circle. The least controlling of the growth rate in the antibiotic was, the control because, it had a 0mm distance. 5. The antiseptics arent as effective of controlling the growth rate of the antibiotics, because a lot of the zone inhibition

was very small. Also, the alchol was the highest in my data, but compared to the antibiotic table, it was the least effective. 6. We know that the inhibition on the disks is caused by the
antibiotics/antiseptics because we dipped the qu-tips into substances and wiped it on our to trigger, plate and our bateria. After we dipped it in, we put them in the incubator to let it grow

Conclusion: My hypothesis was correct because, the tetracycline was the most effective in the inhibition zone. They both had the largest ring and this shows the E-coli was the most effective.