Photos of Maps of Four-Pole Creek

Getting Water Samples from Four-Pole Creek

My dad and I collected water samples from various locations at Four-Pole Creek. We did this to try to determine if bacteria was coming out of certain sewage pipes and if it had the potential of spreading fecal-oral diseases.

Trial Run on Examination of water in Four-Pole Creek for fecal contamination.

Purpose – Study impact of fecal contamination on Four-Pole Creek because of

potential fecal-oral spread diseases.

1. Method selected - MFc - membrane filtration method with incubation at 44.50C to select only mammalian coliforms vs. environmental.

2. Site selected – See map – approx. 50 m below bridge to tennis center.
3. Collected samples in sterile specimen cups with screw-capped lids.
4. Lab – Micro. Cons. - Passed 1ml, 10ml, and 25ml volumes of the sample through a membrane bacterial filter, which was placed on a pad in a small petri dish

soaked with MFc broth. This was incubated in a water bath at exactly 44.50C for 24hrs.

         Results – See photo of plates.  Counting could not be done on a colony counter because the plates were opaque. Instead, pictures of each plate were taken, and the colonies were counted on those.  

Tuesday's Samples

           

 25ml plate:  TNTC (Too High to Count)

            10ml plate: 157

              1ml plate: 16

25ml plate:  TNTC (Too High to Count)

            10ml plate: 144

              1ml plate: 16

            Confirmation that coliforms were fecal in origin:  Test for presence of Escherichia coli,which is found only in feces, and is not found in the environment except in feces.

            Method was to perform glucose fermentation, lactose fermentation, citrate utilization, sulfide production, motility, indole production, phenylalanine deaminase production, and oxidase production.  See photo of those tests, with the results:

Gram Stain Procedure

This was a gram stain procedure I did at Marshall with my dad around the dates February 4-7. He showed me how and what to do. It was interesting. Now I know how to do a gram stain procedure.


Here's what we did and the results:


Performed Gram stain on one G+ organism (Staphylococcus)

and one G- organism (E. coli).  Purpose - identification

1. Prepared smear using aseptic technique.
2. Air dried, heat fixed (gently).
3. Crystal Violet (10 sec.)
4. Iodine (1 min.)
5. Decolorizer (15sec,)
6. Safranin (1 min.)

Results: Staphylococcus appeared as purple spherical clumps, which was expected. The Escherichia coli was a faint pink bacillus, which was also expected.

Streak Isolation Technique

In Mr. McKee's lab I separated bacterial culture into isolated colonies, using a transfer loop and doing a sweeping motion across a plate of trypticase-soy agar, followed by incubation.


Though I don't remember the exact date, we did this I believe on the first week of February.

First Post

I started doing microbiological research about a week ago and will continue doing so until the end of the school semester and possibly into the summer. So far I have learned two procedures for identifying bacteria. I will post details and pictures of them soon. I will also be deciding exactly what research project I will be taking on very soon.

I am excited for being able to do research as well as the end results I might achieve. I've been wanting to do things like this since I was a little kid and I already find it very interesting.

Hopefully this research will be an adventure that myself, those involved, and those reading this blog will find exciting and rewarding!