LAB REPORT 5: SWAB SAMPLE Objectives: 1. To learn the steps how to process a real patient sample in the lab.

2. To detect the disease or infection that the patient was having. 3. To find the suitable antibiotics to use to kill the bacteria infection. Introduction: Swab sample is sample taken from patient using sterile swab. It can be used for skin infection, throat swab, vagina swab, anal swab, ear swab and many kind of swab. After the physician swabbing the patient infected area, the swab was stored in an appropriate container to avoid from contamination of surrounding environment. Then it will sent direct to microbiology lab to be process. In microbiology lab, the swab will examine in many kind of test. It will be observed macroscopically for its characteristics like its colour. Then the swab will undergo Gram staining to see its microscopic characteristics. Then it will be cultured in an appropriate medium that will inhibit the growth of normal flora. Lab technician need to know what the normal floras are and what not in the swab culture based on the colony count on the agar plate so they will not make mistake during the diagnosis process. Last but not least, the colony from swab sample culture need to undergo antibiotic sensitivity test in order to determine which antibiotics are suitable to kill the bacteria and safe to prescribed to the patients. Materials: 1. Swab sample 2. McConkey agar 3. Blood agar 4. Mueller Hinton agar 5. Wire loop 6. Bunsen burner 7. Grams stain 8. Antibiotic Metrodinazole (BAN) 9. Slides 10. Microscope 11. Toothpick 12. Oxidase reagent 13. Filter paper 14. Catalase reagent 15. Peptone water 16. Horse serum

17. Dropper 18. Antibiotics: a) Fucidini acid (FD 10) Method: Day 1;

b) Gentemicium (Gen) c) Erythromycin (E 15) d) Penicillin (Pen)

1. The swab sample cultured in two plate of blood agar and a plate of MacConkeys agar. Then the swab sample was swab on a glass slide. 2. Then the slide was stain using Grams staining. 3. Then it was observed under the microscope and the shape, color of the stain and the organization of the microbes were observed. 4. Then, the cultured sample on the MacConkey agar and blood agar were incubated for 24 to 48 hours in the incubator at temperature of 37C. 5. The other cultured blood agar was inserted with Antibiotic Metrodinazole (BAN) at the second striking and was incubated in CO2 incubator for 24 to 48 hours at temperature of 37C. Day 2; 1. After 24 to 48 hours incubation, the cultured microbes were observed it culture characteristics which in MacConkeys agar are there any bacteria growth or not and if there are growth, it was checked whether it ferment lactose or not. 2. While in blood agar, if there are any growths in it, it was checked whether it was beta, alpha or gamma hemolytic. 3. In the blood agar for CO2, it was checked if there are any clear area around the antibiotics disc. 4. If there are any clear areas around the antibiotics, it indicates that there are present of anaerobic bacteria. 5. Then a small colony from each plate was taken and staining using Gram stain then it was observed under microscope for identification. 6. Then, a few biochemical tests were run to observed it reactions.

7. These include oxidase test, catalase test and coagulase test for Gram positive bacteria. 8. IMViC test, motility test and TSI test for Gram negative bacteria. 9. If there are present of yeast, germ tube test was run. 10. After that, antibiotics sensitivity test were run. The antibiotics used were Fucidini acid (FD 10), Gentemicium (Gen.), Erythromycin (E 15), and Penicillin (Pen.). 11. Then the plate was incubated for 24 to 48 hours in the incubator at temperature of 37C.

Day 3; 1. After 24 hours, the results of antibiotics sensitivity test were measured and recorded. 2. All the results of biochemical test data that were obtained before was compared with the biochemical reactions table that was given earlier and the type of microorganisms was identified. Result: Macroscopic view: The contain pus and yellowish

Microscopic view:

Types = GNC, GPC (1+), GND (3+) and epithelial cell.

Types = Gram positive cocci (Blood agar)

Biochemical test: Oxidase = Negative Catalase = Negative Slide Coagulase = Negative Tube Coagulase = Positive

Sensitivity test:Antibiotics Penicilin G Gentamicin (CN 10) Fusidic asid (FD10) Erythromycin (E15) Discussion: From this experiment, after the swab was stain using Gram stain, the slide that was observed under the microscope. It contains bacteria of Gram negative cocci, Gram positive cocci (1+), and Gram negative diplococci (3+). Including non-bacteria, epithelial cells. This showed that the GND is more compared to GPC. This could showed that GND could be the cause of infection. However more detailed test need to perform to confirm the types of bacteria causing the infection that is culture and sensitivity (C/S). After culture the swab sample on blood agar overnight, the growth colony then was also stained using Gram stain. Under microscope, Gram positive cocci were obtained. Then a few biochemical tests were run that is catalase test, coagulase test and oxidase test. The entire test showed negative result. Then since coagulase slide was also negative, coagulase tube test were done and the result obtained is positive. Diameter of clear zone (mm) 20 21 28 8

Then the isolated colony was cultured on Mueller Hinton agar together with four different types of antibiotics. The result showed that the GPC are most sensitive towards Fusidic acid that is 28mm and less sensitive or almost resistant towards erythromycin that is 8mm. it is moderately sensitive towards Penicillin G and Gentamicin. Conclusion: As a conclusion, from the experiment above the bacteria that might be cause of disease on this patient is Staphlococcus aureus and the antibiotic that can be used to kill this bacteria infection is Penicillin.