Discussion From the results, it shows that there is no inhibition zone present in any of the four Petri dishes.

The inhibition zones around the paper discs are not present. This makes it more difficult in concluding the results since we cannot compare any antibiotic effectiveness towards the bacteria since data cannot be recorded or observed. As introduced earlier, in this experiment, we are using five different types of antibiotics to act upon three different bacteria which include E.coli, bacillus, and staphylococcus aureus. Each antibiotic has its own mode of action or its own mechanism to destroy or inhibit certain bacteria. Take ampicillin for example, it inhibits cell wall synthesis by inhibiting formation of the peptidoglycan cross-link. Streptomycin on the other hand, stops the initiation of prokaryotic peptide chain and induces mRNA misreading leading to mutated bacteria. The other type of antibiotic used for investigating its antimicrobial properties is tetracycline that enables the inhibition of prokaryotic aminoacyl-tRNA binding to the ribosome small subunit. Theoretically, the inhibition zone should be present around each of the discs except for the control disc since it is soaked with distilled water containing zero contaminated substances or minerals. However, after checking the paper discs after leaving in the incubator for 48 hours, we notice that nothing appears around the discs. This could be caused by many reasons which can be explained further. The first and foremost reason for this experiment to be the exact opposite of the theory is due to the temperature of the surrounding and the incubator which alters the results. The Petri dishes are immediately used for experiment without equilibrating it to room temperature before use. Plus, the temperature in the incubator might not be suitable for the bacteria to grow as it may be set too high. The temperature of the incubator during the experiment is 36.5C which is not the average temperature since some bacteria are not thermophilic and can only survive within the range of 32C to 35C. Apart from the temperature, the condensation covers the top of agar surface. The condensation is the result of anaerobic respiration of the bacteria and accumulated on the top of the plate during incubation. To prevent the condensation to accumulate, usually the plates are placed upside down in the incubator. Even though, the Petri dishes are incubated upside down in the incubator but this does not cancel out the possibilities for the condensation to happen. Contaminated culture also one of the reason for not getting any results for the zone of inhibition. When handling the experiment, communication should be forbidden. Water

droplets falls into the agar medium when talking. The droplets could contain bacteria which sometimes more harmful than the one we are using. Furthermore, the forceps might be well sterilized. The Bunsen burner is used to sterilize the tip of the forceps. However, during the study, this step is only done once and the neglection leads to contaminated forceps when placing the discs on the agar surface with bacteria in it. Besides, the agar medium is prepared earlier than the date of experiment. The possibility of the agar medium to be old and expired could actually brings in other bacteria to invade the medium before the tested bacteria is added to the agar culture. To add the agar medium into the Petri dishes, measuring cylinder should be used as to get the same amount or volume in each dish. However, this step is replaced by adding it to half of the height of the plate. This makes the depth of the medium to vary in each Petri dish. In the clinical setting, the standard depth is 4 mm. Plates poured to a depth less than 4 mm will result in false resistant results creating no zone of inhibition around the paper discs. The depth of the agar can affect the concentration of the antibiotic as it diffuses from the disk. The rate of diffusion of the antimicrobial through the agar is dependent on the concentration of antibiotic, molecular weight of antibiotic, solubility properties of antibiotic, pH and ionization, binding upon agar. These factors, in combination result in each antimicrobial having unique breakpoint zone size indicating susceptibility to that antimicrobial compound. In addition, the weakness of zone of inhibition testing itself causes the result to be unidentified. The weaknesses are the microbial growth agars themselves may interfere with the function of some antimicrobial agents. The method has some natural variability, and the zones of microbial inhibition do not always have clear or regular boundaries. Even if the zone is present, the method is not classically quantitative and many other explanation or experiment should be done further.