May 28, 29, and 30 - Day 10, Day 11, and Day 12

Selective and Differenctial Media

Purpose: Isolate bacteria based on their salt tolerance and differentiate among theses isolates for mannitol fermentation.

Materials: Mannitol salt agar plate, blood plate, E and B plate, Phenol Alcohol plate, MacConkey plate and, Unknown bacteria in a slant

Procedure: For each plate:
1. Sterilize the loop and then gathered some bacteria from our original bacteria from the slant.
2. For each slant, Maggie inoculated each plate with a smear from our original bacteria.
3. Then we put each plate into the 30 degrees C incubator.
4. It will remain in the incubator for 24 hours.

Results:
Blood Plate- Some Growth, Gamma Hemolysis

Mannitol Salt Agar Plate- No Growth

Phenol Alcohol Plate- Growth

MacConkey Plate- No Growth

E and B Plate-No Growth

MRSA Test and Strep Throat Bacteria Test

Materials needed: Saline Solution, Swabs, a throat to swab, a nose to swab, Blood Agar Plate,

Procedure: (Strep Test)

1. Get a Blood Agar Plate
2. Swab a throat
3. Spread the bacteria on the plate
4. Put bacitracin on the plate
5. Put the Plate into 37 degrees C incubator
6. Look for lysis of the blood

Results: Much bacteria grew, Alpha Hemolysis


Procedure: (MRSA Test, Nose)

1. Get a Mannitol Salt Agar Plate
2. Dip a swab into the saline solution
3. Swab both sides of the nose
4. Spread what was gathered from the nose onto the Mannitol Salt Agar Plate
5. Spread bacitracin on the plate
6.Place plate into the 37 degree incubator

Results: Bacteria grew yellow in color, therefore positive for S. aureus.

Antibiotic Testing

Materials needed: Antibiotic disks: (Penicillin, Erythromycin, Tetracycline, Neomycin, and Chloramphenicol), Nutrient Agar Plate, Swab, Unknown Bacteria, Flame, Forceps

Procedure:
1. Get a Nutrient Agar Plate
2. Put Forceps into alcohol for purification
3. Differentiate 5 spots in the Agar plate for 5 different antibiotics: Penicillin, Erythromycin, Tetracycline, Neomycin, and Chloramphenicol.
4. Get a swab and gather unknown bacteria from the slant of bacteria and spread the bacteria all over the Nutrient Agar Plate.
5. Put the forceps into the alcohol then into the flame in order to sterilize them.
6. Pick up an antibiotic disk and place on the specified area in the Nutrient Agar Plate.
7. Put Plate into the 30 degrees C incubator
** Repeat Steps 5 and 6 until all antibiotic disks are placed on the Agar Plate.

Results: Penicillin, Erythromycin, Neomycin, and Chloramphenicol all killed the bacteria. Tetracycline, did kill some of the bacteria, but not entirely or as well as the other four did.

Result of Our Unknown Bacteria:

B. subtilis
-Gram Positive
-Bacilli
-Motile
-Aerobic
-Gaseous
-Endospores

Immunodetective  BioKit: Antibody- Antigen Reaction in Agar

Procedure:
1. Get a petri dish divide it into 3 sections
2. In one of the sections make 3 wells and fill them with various solutions:
     Well 1: Goat Anti-swine Albumin
     Well 2: Goat Anti-horse Albumin
     Well 3: Goat Anti-bowvine Albumin
     Well 4: Swine Albumin
2. Replace and and cover the dish at room temperature.
3. Record results after 16-48 hrs

1. Get an Elisa plate and label each section

2. Add antigen (green tube) to wells of microplate strip.
3. Incubate for 5 minutes
4.Wash off the strip with wash buffer
5. Add the serum (yellow tube), positive (purple tube), and negative control (clear tube).
6. Wait 5 minutes

7. Wash it again
8. Use enzyme (orange tube) to detect antigen- add it to each section
9. Wait 5 minutes
10.Wash it again and again
11.Add SUB (brown tube) into each section
12. Wait 5 minutes
13. Observe to see if there is a color change- from colorless to blue. If there is a color change it is positive

Results:
The positive control turned blue meaning it has antibodies towards this antigen (AIDS). The other two controls did not change in color meaning they do not have antibodies towards the antigen.

UV Light
-There is a mixture of all the bacteria in our class.
-Use UV rays to treat it: a UV ray wand was used by stirring and sitting in the beaker of bacteria.

Yogurt
-Boil milk till foaming
-Cool milk to room temperature
-Add spoonful of yogurt to the milk
-Incubate it in37 degrees C