Week 10: Start of Identifying Our Wastewater Bacteria!

 Happy Almost-end-of-March!

This week, the focal point was the starting of the Biolog testing. 

The first step was deciding on two isolates per stage of the wastewater system (since there are 5 collected stages, we selected 10 isolates in total). 

Our selection process was primarily based on how unordinary the bacteria species grew on the general media (I noticed I  preferred orange bacteria because of their unique color). Once we determined the 10 isolates we wanted to test, Eveyln and I inoculated the bacteria from the patch plates (grown by the microbiology students) onto TSB under the fumehood and incubated for 24+ hours. 

We had a laboratory and environmental control (Stage 3, Isolate 3). 

The laboratory control was Escheria coli. Using a sterile loop, I inoculated an E. coli-containing broth onto BUG (Biolog Universal Growth) agar to be incubated as well. 

Once I saw growth on the BUG agar, I then inoculated that bacteria onto another broth (IF-A) to begin the metabolic profile testing using the Biolog. 

We measured 100 microliters of the IF-A broth with the E. coli and placed that volume into each weir of the microplate. Each weir consists of a specific substrate that the Biolog will read to determine if the bacteria is metabolizing it or not, and from that information, the Biolog matches our environmental isolates to their microbiology database!

I let the microplate incubate for 24+ hours before reading it with the Biolog system. 

The control read by the biology successfully identified the bacteria as Escherichia coli!

The exact process for environmental control was executed, and the Biolog was able to identify the bacteria as Aeromonas eucrenophila.  

With the controls providing a green light to a functioning and happy biology machine, we can now repeat these processes for each stage!

I attached a screenshot of the Biolog results for the environmental control (Stage 3, Isolate 3) below!