I am a college student studying microbiology. My peers and I just
recently performed an experiment where we tested hamburger meat for
bacteriophages using different strains of E.coli. The procedure we used was
the hydrogen peroxide treatment with crystal violet agar plates for
selective recovery of bacteriophages. For our lab report we would like to
compare our results and procedures to ones that might have been possibly
done in the past. We have tried the internet, library, and the Journal of
Applied and Environmental Microbiology. So far we have had next to no luck
finding any articles that pertain to this test and food. I was wondering if
anyone might know of a source that we might not have tried or a experiment
that was done that was similar to ours.
Here is a detailed explanation of our procedure with a reference for
anyone who might not be familiar with it. First we inoculated 10mL of TSB
with five different E.coli host strains. We then let them incubate for 24
hrs in a 37 degree celsius water bath. The next day we took 50g of a meat
sample, and added water to bring the sample up to 100ml. We added 3.3mL of
3% hydrogen peroxide per 100mL of sample. We then shook them for one hour at
room temperature. After the hour period, we inactivated the peroxide by
adding 5mg Catalase per 100mL. We took five different meat samples. We
added 100ml of water and then blended to liquid. We then added 0.1 mL of the
E.coli host and 0.1 mL of the sample into 4mL top agar. The top agar was
then mixed and poured over crystal violet plates. The plates were then
incubated at 37 degrees Celsius for 24 hrs and were checked after 8 hours
and at 24 hrs.
Our results were inconclusive because no phages were found. A similar
experiment was performed last year at St. Francis and phages were found. We
are trying to find a new procedure or similar ones to try to explain our
lack of data. Our professor would like to see us find as much information
as possible so that we can put together a report for next year's students to
research. Unfortunately, all we can find is the recovery of phages from
water samples and sewage. Very little has been recovered so far an solid
sampling.
The experiment was derived by my peers and I using an experiment that was
in The Journal of Applied and Environmental Microbiology. Issue is April
1992 p 1159-1163 title is Use of Hydrogen Peroxide Treatment and Crystal
Violet Agar Plates for Selective Recovery of Bacteriophages from Natural
Environments. It was published by Abdolkarim Asghari.
Thank you,
Michelle Kerr
Sophomore at the University of St. Francis
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