Microbiology Lab Report

Christopher Baaklini (Biochemistry 2nd year English)

ID: 57396

ANTIBIOGRAM
PRINCIPLE:
Antibiotic discs of known concentrations are impregnated into the Muller-Hinton agar which diffuse into the
gel creating a concentration gradient. MIC is then determined by measuring the zone of inhibition around each
disc. Resistance characters of the bacteria will be deduced.

METHOD:
- Prepare in advance a bacterial suspension from an isolated colony inoculate in a nutrient broth.
- Take about 1 ml of this suspension using a Pasteur pipette and spread it on Mueller Hinton agar.
- Let the inoculated suspension dry for 1⁄2 hour in the incubator.
- Dispense and fix the antibiotic discs in a symmetrical way using a sterile forceps.
- Incubate for 24 hours at 37 °C, then read the results.

RESULT:
The bacteria studied is resistant to oxacillin as no zone of inhibition was observed around the respective disc.
The other four discs show zones of inhibition of different sizes with chloramphenicol 30 having the largest zone and
carbenicillin the smallest.
SPREADING
PRINCIPLE:
Check the presence of bacteria and visualize them by spreading a given mixture on the agar.

METHOD:
Form the glass rod into an L-shape using heat from the Bunsen burner.
Using a dropper put 2 drops of the mixture onto the agar.
Spread the mixture over the agar using the L-shaped rod.
Incubate and observe after 24 hours.

RESULT:
An important growth of bacteria is observed on the agar indicating it was present in the mixture. The extremely
small bacteria colonies formed seem to have a circular shape with a creamy consistency.
BACTERIAL IDENTIFICATION
PRINCIPLE:
Identify bacteria according to their biochemical characteristics. Tests are carried it with pure bacteria.

METHOD:
Using an inoculation loop, a small sample of bacteria is taken and spread in 3 different test tubes containing
Simmons Citrate, TSI (Triple sugar iron agar), and Mannitol Nitrate tests.

RESULT:
1. Simmons Citrate:
Green color persists in the tube indicating that the bacteria is citrate negative

2. TSI:
A homogeneous yellow color is observed throughout the tube indicating the fermentation of all 3 sugars;
glucose, lactose, and sucrose.
Lack of black color indicates absence of H 2S.
No bubbles indicate absence of CO2.
3. Mannitol Nitrate Motility test:
A yellow color is obtained indicating the fermentation of mannitol present in the medium. Thus the
bacteria are mannitol positive.
Additionally, visible growth away from the stab line indicate motility of bacteria.