Name: Ruhi Kiflen Student ID: 999807641 Lab Section: P0401 Lab Assignment: Pre-Lab 1 Name of Lab Partner

: Qixing Da 1. Materials and Methods [21 marks]: Obtaining the Daphnia Magna and its validity This experiment was designed to test the effects on the heart rate of Daphnia magna when exposed to different concentrations of ethanol solution. Performed on October 10, 2013 in a closed environment at the University of Toronto, the Daphnia magna were collected from the Cell and Systems Biology (CSB) Daphnia Colony. The Daphnia sp. are a relatively common model organism that survive well in culture. Furthermore, they are able to quickly adapt morphologically, physiologically, and behaviorally during environmental changes ensuring their validity as a model organism. (Appendix 3) Preparing the Solutions To study the influence of the ethanol, 10% ethanol was used to prepare different concentrations at 2%, 4%, 6%, and 8% ethanol solution in 5 petri dishes. Measuring Baseline heart rate 6 animals were tested with each concentration ranging from 0% ethanol to 10% ethanol. To obtain the heart rate, the Daphnia magna were transferred with the use of a plastic wide-mouthed pipette to safeguard the species from damage. The species were placed onto a depression slide with a few fibers of cotton wool and a small volume of bathing solution (5-10 l). These methods were used to ensure the immobilization of the Daphnia magna. To ensure the ability to clearly and reliably count the heart rate, the slide was placed on a Daphnia magna cooling chamber with ice. Each animals heart rate was recorded three times at 10-minute intervals to reduce stress levels and establish a base line, each for a period of 20 seconds under a stereomicroscope and light. To maximize

results when counting, the number of heartbeats was dotted with the use of a sharpie and filter paper. Exposure to ethanol and recovery period The first treatment solution was added. After a wait time of 3-5 minutes which gives the Daphnia some time to adapt to the environment, the heart rate was observed three more times at 10-minute intervals. This process was repeated until all the animals were tested under each concentration. To remove the treatment, Kim wipes were used. The species were allocated in a Daphnia magna rehabilitation chamber for full stability of the animal. Data Analysis Prior to analysis, the heart activity was converted to beats per minute (bpm) through the multiplication of each heart rate obtained by a factor of 3. The heart rate over various concentrations was examined by a bar graph. A t-test was used to compare the differences in heart rate at the various concentrations and the p-value was used to determine whether these differences were major. To analyze the data, a paired t-test was utilized because data was collected after the manipulation of the heart beat count.

2. Hypotheses a. List your Hypothesis After adding ethanol at different concentrations, I expect the heart rate of the daphnia to decrease as the concentrations of the ethanol increase. b. Cite one peer-reviewed manuscript that provides a rationale for your prediction Corotto, F, Ceballos D, Lee A, Vinson L. 2010. Making the Most of the Daphnia Heart Rate Lab: Optimizing the Use of Ethanol, Nicotine & Caffeine. Ame. Bio. Teach. 72 (3): 176-179. c. Write two to four sentences explaining to your TA why you chose this manuscript. Ensure that you indicate why it supports your hypothesis This manuscript was chosen because it examined the effects of ethanol on the heart rate of Daphnia, which is what Id like to examine and observe in my lab. To see how another group of people have done the same experiment helps to bring perspective on new things that could be tried to see how it would affect the results. It supports my hypothesis because the results showed a decrease in the heart rate with the use of ethanol.