NyQuil and its Effects on Paramecia

       One of my more interesting courses I'm enrolled in this semester is Genetics and I think most of the interest comes from the lab portion of the course curriculum. In particular the Paramecia lab that we just finished. The goal was to induce a mutation onto a culture of Paramecia but, the methods and materials were up to us to determine and experimentally figure out. My lab partner and I chose NyQuil cold and flu liquid to be our mutagen becuase my partner frequently uses NyQuil for her colds and flu symptoms so we wanted to see what it would do. We introduced the Nyquil to our culture of Paramecia at a variety of concentrations that were determined after a few test trials.

       Paramecia are generally studied for observation of their cilia on their outer membrane and their simple lifestyle. They are single-celled organisms that live in fresh water environments such as ponds (Genoscope, 2007). Paramecia are on average 120 micrometers in length and covered in hair-like projections called cilia, which are easy to observe under a light microscope in a laboratory (Fraga, 2001). The cilia on the Paramecia allow the organisms to swim in one direction, under normal conditions they swim only forwards but, when under toxic or abnormal conditions they reverse and swim backward to try to find a homeostatic environment (Fraga, 2001). When they are in a normal environment calcium channels are closed in the ciliated membrane so the concentration in higher outside the cell than inside the cell (Genoscope, 2007).When they are under abnormal conditions the calcium channels open and the calcium ions flush into the cell to reach an equilibrium between the outside environment and the inside environment (Genoscope, 2011). Once regular calcium levels are restored the calcium channels are closed again and Paramecia swim forward again.



Figure 2: Experimental wells
with NyQuil concentrations 

        The Paramecia culture was made with a wheat media and inoculated with the bacteria Klebsiella pneumoniae as the food source for the Paramecia. A flask containing 50mL of wheat media was inoculated with the bacteria and left to incubate for twenty-four hours, after the incubation period 1mL of Paramecia cells were added to the flask and left at room temperature to grow and multiply for a week. The prime time for experimentation on the Paramecia cells is 6-8 days old, this is when they are most healthy and abundant before they start to die off and become less viable. To keep a viable culture of Paramecia we remade cultures of cells each weeks for the following lab period.

Figure 2: Control Paramecia.

        Once the week was over the experimentation began. We did simple calculations to determine concentrations of NyQuil to Paramecia, starting with 50%, 10% and 5% solutions. After seeing that a 50% solution of Nyquil to Paramecia left the cells stunned and 100% dead we knew the concentrations we were looking for were much less than that.

       In the 10% solution the cells were still alive when transferred to a slide for observation. These cells had very different swimming movements, almost drunk-like. They were spinning and barrel-rolling around in the solution, after several minutes elapsed of the erratic swimming the cells started to slow down and eventually stop moving entirely. The cells seemed to have a slight change in shape and appearance as well as the abnormal swimming patterns, as seen in the photograph of the control cell to the cells in NyQuil solutions there are several morphological changes.

       These changes couldn't be defined as mutations in the actual genetic code or just situational mutations because the morphed cells didn't survive overnight to be transferred to a new culture medium. The 10% solution cells died rather quickly so we tried 5% which caused the same changes both behavior and morphological but again these cells didn't survive overnight. The next week we tried solutions with concentrations of 4%, 3%, 2%, and 1% NyQuil. With more time to keep these
cells in these concentrations we saw the cells start to appear to be exploding their membranes. In the picture the the left, you can see the bubbled out  of the membrane of the cells displaying the lysis process of cells. The behavior and morphological changes seen in the previous concentrations were also seen in these solutions but, with the decreased concentration of NyQuil the cells were swimming erratically for longer and longer periods of time before their swimming got to be sluggish.

        I think that if we had more time to experiment with these Paramecia we could have gotten a concentration more exact that would have sustained the life of the cells in the solution so we could have concluded that an actual mutation had taken place. Although I think we made some kind of change with the shape of the membrane of the cells as well as the swimming patterns there is no statistical data that we caused an actual mutation to the Paramecia cultures.

          References

Fraga, D. (2001, July 2). How ion channels control paramecia behavior. Retrieved from http://www3.wooster.edu/biology/Ciliates/citc/Para_behavior.html

Genoscope. (2007, Sept. 11). Paramecium teraurelia. Retrieved from http://www.cns.fr/spip/Paramecium-a-model-ciliate.htm

** All photos taken by myself (Abby Chauvin) in the lab using Image J software**