User:Katelyn R. Porter/Notebook/0703, T4, transect protist observation

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Identifying Protists and Plating Serial Dilutions


Purpose: To practice identifying unknown organisms, to examine protists and algae from a transect, and to prepare and plate serial dilutions from the transect.


Materials and Methods:

-A wet mount was made of a sample containing several organisms and was observed with a compound microscope to practice identification

-A sample was taken from the farm hay infusion culture to create a wet mount for microscopic observation

-100 µL from the farm hay infusion culture was put into a 10 mL broth tube to make a 102 dilution

-100 µL of that broth was added to a different 10 mL broth tube to make a 104 dilution

-The above procedure was repeated to create a 106 and 108 dilution

-100 µL of each dilution was spread on an agar plate and tetracycline plate to create four dilutions of 103, 105, 107, 109 each on an agar plate and tetracycline plate


Observations:

The practice protozoa observed was large, colorless, and ovular in shape. It exhibited apparent motion using flagella and when it moved it became elongated with a broad, rounded posterior and appeared to be vibrating when in motion. Using a dichotomous key, this was classified to be of the Peranema sp.

The hay infusion culture had a strong smell to it and there was no apparent life on the surface of the liquid, only soil. Organisms may differ close to versus away from any plant matter in the culture because an organism may or may not utilize the by-products of plant metabolism or the other organisms that live around plants. If the hay infusion culture had been observed for another two months, there would have been more organisms growing and more variability in species.

The first potozoa observed was circular, about 60 µm in diameter, colorless, and crept slowly used cilia as its organ for motion. Because it was colorless, this protist was non-photosynthesizing. Its shape remained constant and it possessed a flattened shell without attached material, which was a pale brown color. This protist was classified as an Arcella: Image:Transect_uknown_protist.jpeg‎

The second protozoa observed was colorless and about 40 µm long. Since this protist was also colorless, it was non-photosynthesizing as well. It appeared to use two flagella for motion, moving rather quickly, and was elongated with a narrow posterior. This organism was classified to be of the Chilomonas sp.:

Image:Transect_unknown_protist2.JPG‎


Serial Dilutions:

Image:Serial_dilutions1.JPG‎

Example of plate spread:

Image:Serial_dilutions2.JPG‎

(100 µL of the dilution was used for each plate)

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