Wilson Huete

1/18/2019

Purpose: The purpose of this lab was to become more accustomed to reading and classifying scientific literature.  The article we received from Dr. Adair was The D1-D2 region of the large subunit ribosomal DNA as a barcode for ciliates. This was classified as a primary source.

Goals: To earn an A in the class. To learn more about ciliates and learn more about research and how to use it in my undergraduate career.

Wilson Huete

11/16/2018

Purpose: The purpose of this lab was to identify what kind of soil we collected through the identification of percent soil composition. This is important because certain ciliates are only found in certain environments. We also continue to search for ciliates in our non-flooded plates.

Procedure:

1. Collect your falcon tube.
2. Observe the different layers that have formed and use a ruler and take a picture so you may calculate your percent composition.
3. After finding out your percent composition use the chart to identify what kind of soil you have.
4. When you have obtained that information return you falcon tube to the rack
5. Obtain your none- flooded plate
6. Use a pipettor to place 3 20ul drops on a slide
7. Then using a compound microscope search for ciliates and when found take a picture
8. Then use a 3ul drop of methylcellulose or iodine to stain the sample to make the ciliates stop moving.

Data:

Conclusion:

through the soil composition analysis, the type of soil I had silty clay loam. I have consistently been finding ciliates everylab.

Future Steps:

My future goal is too go somehwere else in a different kind of soil and cllect a sample and see what possible ciliates could be found in that kind of soil.

Wilson Huete

11/9/2018

Objective:

The objective of this lab was too further analyze our soil samples and to continue trying to identify ciliates. Another objective was to learn how to classify soil through its different compositions.

Purpose: The purpose of this lab was to continue trying to find ciliates in our soil samples.

Procedure:

1. Obtain a clean falcon tube and fill with soil until it reaches 4ml
2. Then using 6ml of water filling the tube until the 10 ml mark
3. Add 1 drop of dispersing solution
4. Use the spin vortex for at least 30 second
5. Mark the tube with your initials and group number and place on the rack in the front of the class.

Ciliate observation:

1. Place a 25ul drop on a slide
2. Search for ciliates and if found go to a higher power on the compound microscope
3. When found you can use iodine drop or methylcellulose to slow down the ciliate
4. Take picture of the ciliate.

Data:

This picture was taken by Christina Clark my lab partner

Conclusion:

At the end of this lab, I was able to see how ciliates were able to be dormant for extended periods of time but they can come out of dormancy. This lab also helped me further my microscope skills as trying to locate minuscule ciliates is very complicated.

Future Steps:

In the future, I would to keep studying ciliates and see what other things these organisms are capable of.

Wilson Huete

11/2/2018

Purpose:

The purpose was to determine the soil ciliate diversity in the soil samples previously collected.

Data:

Mass of Empty Petri Dish

5.8g

Mass of Petri Dish + wet soil

30.8g

Calculate the mass of the wet soil

25g

Mass of petri dish + dry soil

22.8g

Calculate the mass of dry soil

17g

Procedure:

Percent water lost:

1. When you first collected the soil it was weighted in the Petri dish with and without the lid.
2. After a couple weeks the soil was reweighted without the lid.
3. we can calculate the percentage of water lost through the equation : (wet mass-dry mass)/(wet mass) x 100= percent water loss.

PH Of Soil:

1. Remove 1ml of liquid from the soil to a microfuge tube.
2. Microfuge it for 15 seconds
3. Obtain a strip of ph paper(5.0-9.0) and submerge in the tube and see the color change.
4. Compare the strip to the key and determine ph.

Ciliate Hunt:

1. Scrape the dirt to one side of the Petri dish and use a dissecting microscope to look for ciliates.
2. When seen use a pipettor to pick up the ciliates.
3. Transfer to a clean slide.
4. Using a compound microscope observe and look for ciliates.
5. If ciliates move to fast use iodine or methyl drops to slow down.

Results:

Percent water lost: 32%

PH:6.5

Conclusion: I was able to observe ciliates but when trying to take a picture I was unable to take a clear one. This experiment helped us understand ciliates better due to the fact we were able to observe them in their own environment and living conditions.

Future steps: With the completion of this lab I hope to understand ciliates better and further explore them.

Wilson Huete

10/26/2018

Objective/ Purpose:

The purpose of this lab was to help us in writing our scientific research paper as well as to help us to better format our research rough draft that we will present.

Sections Of The Paper:

Titles And Authors:

The title should be long and should help give the reader what was researched and or tested. In the author’s section everyone who helped be mentioned, however only your group should be listed as authors.

Abstract:

The abstract section should be no more than 250 words and it should give the reader a summary of the whole experiment it should also include certain information such as small snippets of important information, hypothesis, data, conclusion. At the end of the section, the reader should understand how the results were obtained and what they mean.

Introduction:

Should include at least 3 primary references (APA format). The introduction needs a specific title that should catch a readers attention. The purpose of the introduction is to go into further detail in the experiment as a whole as well as relevant background information.

Materials And Methods:

This should be a concise and explicit version of everything you did in the experiment. It should be written to the point where anyone could recreate the steps you did exactly. Should be written in past tense.

Results:

This should include all your figures as long as a small description for the reader to understand what the data shows. There should not be any explanation in this section.

Discussion:

This is where you describe the results of all your figures. You tie everything together and you explain if the data supported your hypothesis and if it did why. Also talk if any errors may have occured. Also talk about outlieres.

Future Goals:

I hope to be able to write my rough draft and over time become more precise in writing them as well as continuing research.

Wilson Huete

10/12/2018

Purpose/ objective: The purpose of this lab was to familiarize oneself with excel and be able to create a histogram, f-test, t-test, and descriptive statistics.

materials: excel with analysis pack

Procedure:

1. Open a downloaded excel from canvas that has the section 34 data
2. Place all the treatment data for cell counts in a column and do the same for the control as well
3.  Then obtain the descriptive statistics for it
4. Then create a histogram for both the control and treatment you will need to create bins for both of them and it will be trial and error making those
5. Then you will do the f- test and variance 1 has to be the one with the greater variance.  the f test is for both treatment and control.
6. After that, you will do a t-test doing the same thing
7. Repeat steps 2-7 for swim speed as well.

Data:

Cell Count:

Swimm Speed:

conclusion:

Through the analysis of all the data the null hypothesis has to be rejected and the mean for treatment group is higher than the control.

Future Steps:

My future steps is to be able to become more familiarixed with excel and see how time furthers affect the treament group of tetrahymena.

Wilson Huete

10/05/2018

Objective:

The objective of this lab is to compare the absorbancy between the control and treatment group as well as doing your assay with them.

Purpose:  Is to do your assays with the control and treatment groups.

Materials:

• sterile glass jar
• .5g of hay bailing twine (the brown one)
• ppt that was microwaved for one hour and stored for 168 hours

Methods:

Spectrometer:

Absorbancy was tested for both the control and treatment group at 600nm

Cell Count:

You take 3 drops of both the control and treatment group in 2ul drops and you fix them with a 1ul drop of iodine and you count the number of Tetrahymena in each drop and record them.

Swim Speed Assay:

You take a 2oul drop of both groups in two separate flat slides and put them under a dissecting microscope. You place the camera on the microscope and record both samples while you have a ruler there so you can time how long it takes 10 cells for both groups travel one millimeter.

Megan did the direction change.

Christina did the vac assay

Data:

swim speed assay

Storage: All microscopes were put up and all slides used were cleaned and stored for reuse.

Future Steps:

I hope to be able to see what other experiments we can perform with Tetrahymena as wells if any other assays could be used for our experiment.

Wilson Huete 34

9/21/2018

Objective: To learn how to 100p pipettors and also how to dilute ppt and to start planning out your research experiment.

Procedure:

1. Go to canvas find and follow the instructions on how to collect a soil sample depending on what section you are in.
2. Pick where you want to collect your soil sample from the area you are assigned.
3. Take pictures of the tree, the tree leaves, the soil, and the surroundings of the tree.
4. Get your sandwich bag and using a plastic spoon obtain the soil sample which should fill up the bag up to half( begin collecting at 6in of depth).
5. complete everything required online
6. Obtain an empty Petri dish and cover the side where you view specimen with dirt and using a sharpie label the petri dish also label the sandwich bag.
7. Before you fill the petri dish with soil measure the weight and also measure the weight after the soil has been added.
8. Obtain a 1000p pipettor and practice obtaining and releasing liquid with water.
9. Obtain a 24 well plate with a stock solution of Tetrahymena.
10. Use a dissecting microscope to see if any ciliates are alive in the stock solution.
11. Using the 1000p transfer 900ul of media to 4 wells in the 24 well plate
12. then using a 200p transfer 100ul of stock to that one well which make a dilution of 10^-1
13. Then obtain a 100ul of solution from the 10^1 well to another well which would make it a 10^2
14. repeat this step to keep diluting until you reach a dilution of 10^-4
15. Using a compound microscope and a concave slide using the 10p drop a 5ul drop of the 10^-1 dilution and observe under the microscope.
16. repeat this step two more times.
17. calculate the cell/mm.
18. Go to the computer lab with your group so you can start planning the research experiment.

Data/ observations:

Mass of empty petry dish: 5.7g

mass of full petry dish: 34.1g

Equation: (number of cells)/5ul X dilution factor X 1000

We went to the computor lab and were discusing how to set up our expirement and what was going to be our expirement. My group decided to test if the concentration of microplastics affacted the survival rate of Tetrahymena. Our hypothesis is If microplastics affect the survival rate of tetrahymena then the higher concentration of microplastics will decrese the survival rate of tetrahymena.

Storage: the petry dish was labbeld WH34F18 and the rest of the soil sample was labled the same, however the petry dish was stored in the fume hood and the rest of the samples were put in plastic bins.

Conclusion: The expiriment me and my lab partners designed seems testable. As the experament continues we will see if our procedure works and if the experiment is testable. Diluting was an important part of our experement and more pipeting will also be crucial for our expirement. This lab has helped us further develop important lab skills.

Objective: The objective of this lab is to give students the opportunity to familiarize themselves with how to work a dissecting microscope, and how ciliates behave. Another objective is for students to be able to identify a different kind of ciliates.

Procedure:

1. obtain a clean well plate
2. obtain 6 clean pipets and the 6 specimens
3. using the pipets transfer the specimen to an individual well
4. Use the dissecting to observe all 6 specimens and write down observations
5. Sketch what the ciliates look like and identify them using the information provided by the data sheets.
6. repeat steps 1-5 for all samples
7. pour bleach solution into well plates and rinse thoroughly leave upside down on paper towels so they can be dried and clean up lab area completely making sure all equipment used is put up correctly.

Conclusion: I believe my partners and I identified all the ciliates correctly we could not identify the first one due to the fact it was extremely small and hard to see. Something interesting I saw is how all the ciliates have the same organelles to move however, the shape and length make them move in many different ways and at different speeds.