MUIH Microbial Analysis Experiments

Spring 2018 I began my internships. I wanted at least one of them to be focused on scientific testing and lab work since I have a keen interest in the science behind the plants I study and work with.  To meet this goal, I did two formal internships with MUIH faculty, on the Microbial Analysis Experiment.

The Microbial Analysis Experiment was a joint experiment between the University of Maryland’s Joint Institute for Food Safety and Applied Nutrition (JIFSAN) and the Maryland University of Integrative Health (MUIH) Dispensary.  The research objective of the overarching study was to compare the results of microbial testing conducted in a laboratory setting versus multiple non-laboratory settings that use a bunsen burner to create a sterile field. The study was broken into 3 phases, with my first internship occurring during phase 1: the Research Phase.

My assignment was to conduct research remotely to: determine the regulatory requirements for microbial testing, develop the steps for selecting and preparing a laboratory environment in a remote location, and develop a list of materials that would be required to conduct microbial testing in an aseptic home/remote lab.  All work was due by April.

I conducted an initial literature review and evaluated the existing body of research. Each of the sources in my literature review provided information as to industry standards for testing or how to properly prepare a lab environment.  This research provides small business owners with the education necessary to properly test botanical matter. The sanitization techniques are also applicable for product development and manufacturing.

Phase 2, the Bunsen Burner Experiment and Phase 3, Microbial Testing were both conducted Summer 2018, however they were non-credit internships. I had already signed up for a separate internship for Summer 2018 (working at the U.S. National Arboretum National Herb Garden), however I was excited about the prospect of participating in a study between two universities, with the possibility of the findings being published so I did it without receiving academic credit (I got credit as a contributor). There were two distinct parts of this phase of the experiment: the bunsen burner experiment and the echinacea testing.

We were testing the theory that a bunsen burner could create a sterile field. The reason this would be cool is because many product makers are small businesses that cannot afford to rent lab space for product creation.  The execution of the experiment required learning and using aseptic lab techniques (limiting variables), meticulously documenting each step and then counting colonies.

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The bunsen burner was turned on for five minutes to create the sterile field.  Measurements on the table indicate the distance of each TSA plate (a.k.a. a petri dish to most) to the burner.  After 5 minutes the lids were removed to expose them to the microbes in the air.
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After the predetermined amount of time the lids were replaced and the dishes were wrapped in lab film to keep them secure.
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Close up of the dishes before the incubation period.
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Here is a couple of days in.
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This is after 7 days of incubation. Look at all of those organisms!

Results of the experiment were presented at the 2018 MUIH Research Symposium.

The last portion of the experiment I participated in involved testing botanical extracts against various bacteria, yeast, and mold.  Specifically, I got to test hydro-alcoholic extracts of Echinacea purpurea (commonly known as Purple Coneflower or Echinacea), of varying strengths to see if it inhibited the growth of the bacterias, yeast, and mold.  To do this, I created dilutions using 3 different Echinacea tinctures to inoculate, incubate, and interpret results from 3M Petrifilms.

I learned aseptic lab techniques over the summer doing the first part of the experiment. This time around, it was even more important because I was handling petrifilm loaded with yeast and mold spores, and one with E.Coli (yuck!).   Having the experience gained from the first round made it much easier to confidently carry out the steps while limiting exposure.  Some of the additional daily tasks involved taking ambient temperatures, monitoring samples, counting colonies, and reporting results.

The results of this work were combined with later testing to present at the 2019 MUIH Research Symposium.

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