October 11

Tasks for the day:
  1. Culturing Anaerobic Bacteria by using Thioglycollate Broth and the GasPak Anaerobic System.
  2. Learn how to preserve bacteria


Two systems were used to test what conditions my unknown K bacteria grows in.

Purpose:  Does my bacteria use oxygen to grow? If it does then does it need a little oxygen to grow, or does the bacteria simply tolerate oxygen? Or not?

OVERALL RESULTS: My bacteria is facultated because it grew in the presence of oxygen and without.

 THIOGLYCOLLATE BROTH

Materials:

Thioglycollate Broth tube
 Bacterial culture in slant

Procedure:
1. Transfer bacteria from the slant to thioglycollate broth using a sterile inoculating loop.
2. Place broth in the incubator at 37 degrees C for 48 hours.

Thiglycollate broth pre contamination

Below is a photo of the thiglycolate broth after 48 hour growth in the incubator at 37 degrees C.

Note the three layers of bacteria growth.

1. There is a thick layer on the top  - Lots of oxygen present.

2.Lots of growth in the middle -some oxygen present.

3. Growth in the lowest levels - no oxygen present.

     

GASPAK ANAEROBIC SYSTEM 

Materials:
Unknown bacteria slant
Inoculating loop
New agar plate
Anaerobic jar with lid and screw clamp
GasPak envelope
distilled water
Pipette, 10 mL
Pipettor
GasPak anaerobic indicator strip in a packet
agar plates, inoculated with bacteria

Procedure: 

With the inoculating loop take a sample from unknown "K" bacteria and streak on new petri plate. (For the experiment I shared half of an agar surface with another group to save space.)

Place the inoculated plates agar side up in the anaerobic jar.

Contaminated agar Plate ready 

GasPak

GasPak bacteria results after 48 hours

-NOTE Dr. Pathakamuri completed the rest of the experiment by himself- 

Cut off one corner of the GasPak envelope and place the envelope in the jar. Add 10 mL of distilled water to the envelope.

Open the packet with the indicator strip and place the strip someplace visible in the jar.

Secure the lid on the jar with a screw clamp.

Place the jar in the incubator at 35 degrees C for 48 hours.  (within two hours the strip should have no color. This indicates that no oxygen is present in the jar)

PRESERVING BACTERIA

Task: To preserve bacteria for years to come

Materials:

300 m glycerol - used to protect bacteria
700 m Bacterial broth

Procedure: Into a little capsule place 300m glycerol and 700m bacterial broth. Place in a freezer at -80 degrees C.

Tool used to measure broth and glycerol

Taking contaminated bacteria broth

finished product of preserved bacteria

CANDLE DEMO

This was done to show us how to create a jar with no oxygen.

Simply take a jar, place a candle in it lit. Close the jar and wait until the flame goes out.
By the flame going out = no oxygen is present anymore.

Place lit candle in jar

Close jar and wait

No flame, no oxygen