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Biology Lab Report

NAME NRIC GROUP STUDENT ID TITTLE OF PRACTICAL DATE LECTURER

: : : : : : :

NURAINA SAKINA BINTI ZAKRI 951010-11-5076 11SC2 1311171218 THE EFFECT OF CAFFEIN ON HEART RATE 1ST OCTOBER 2013 MR WILLIAM NGU

THE EFFECT OF CAFFEIN ON HEART RATE OBJECTIVE To investigate the effect of caffeine on the heart rate of Daphnia sp. (water fleas) To highlight experimental and investigative skills

INTRODUCTION PROBLEM STATEMENT What is the effect of concentration of caffeine on the heart rate of water fleas (Daphnia sp.)? HYPOTHESIS There is a relationship between the concentrations of caffeine on the heart rate of water fleas (Daphnia sp.). In this experiment, the greater the concentration of caffeine, the higher the heart rate of water fleas (Daphnia sp.). VARIABLE Independent variable Dependent variable Constant variable : : : The concentration of caffeine solution The heart rate of Daphnia sp. Temperature and pH of the environment of Daphnia sp., time to record the heartbeat of Daphnia sp., the type of Daphnia sp. used APPARATUS Pippeters, cavity slides, scientific calculator, spatula, weighing dish, electronic mass balance, 250 ml beakers, 100 ml measuring cylinders, glass rod, stopwatch and light microscope. MATERIALS A culture of Daphnia sp., filter paper or tissue paper, distilled water or pond water, caffeine powder (coffee powder) PROCEDURE

1. 5 beakers containing 100 ml distilled water is prepared by measuring 100 ml distilled water using a 100 ml measuring cylinder and placing it in a 250 ml beaker. 2. Using an electronic balance, caffeine powder of mass 0.1g, 0.2g, 0.3g, 0.4g and 0.5g is measured. Firstly, an empty weighing dish is placed on the electronic balance and the mass of the weighing dish is noted. Then, with the weighing dish still on the electronic balance, the measured mass is set to 0g. After that, by using a spatula, a small amount of caffeine powder is placed inside the weighing dish and the desired mass of caffeine powder is obtained. 3. To prepare 0.1% caffeine solution, 0.1g of caffeine powder is dissolved in 100ml distilled water. A glass rod is used to ensure that all the caffeine powder has dissolved. This step is repeated by dissolving caffeine powder of mass 0.2g, 0.3g, 0.4g and 0.5g into 100ml distilled water respectively to obtain caffeine solutions of 0.2%, 0.3%, 0.4% and 0.5%. 4. A light microscope is set up. A low power objective lens of 40 magnification is used. 5. One single Daphnia sp. is selected from the Daphnia sp. culture. By using a pippeter, the selected Daphnia sp. is carefully sucked out of the culture and then transferred to a cavity slide along with an appropriate amount of pond water. 6. By using a filter paper, the excess water on the cavity slide is absorbed so that the Daphnia sp. lays on its side and has limited movement so that it can be easily viewed under the microscope. However a small amout amount if water is left so that the Daphnia sp. can survive. 7. The cavity slide is then place on the microscope stage and held in position using stage clips. 8. The microscope is adjusted by first adjusting the coarse focusing knob and the fine focusing knob until a fine image of the Daphnia sp. is observed. The position of the cavity slide is adjusted until the heart of the Daphnia sp. can be seen clearly. 9. .One student from the group is assigned to observe the Daphnia sp. using a microscope and count the number of heart beat or leg beat made by the Daphnia sp. within 15 seconds. A scientific calculator is programmed so that every successive tapping of the = button records one heart beat or beating of legs. Another student is assigned to record the time by using a stopwatch. This step is repeated again to obtain a second reading. The average of value of the readings is worked out.

10. The Daphnia sp. is then returned to a beaker containing the same pond as the Daphnia sp. culture. 11. Then another Daphnia sp. from the Daphnia sp. culture is obtained using the same methods in step 5 and 6. 12. A few drops of caffeine solution of concentration 0.1% is added to the Daphnia sp.. The Daphnia sp. is left in the caffeine solution for 1 minute. After that, its heartbeat is counted by the same student and same methods as in step 9. 13. Steps 10 to 12 are repeated by using caffeine solutions 0.2%, 0.3%, 0.4% and 0.5% respectively. 12. Each average value of heart beat of Daphnia sp. is multiplied by four to obtain the value of heart beat of Daphnia sp. in unit of beats per minute. 13. The results of the experiment are tabulated in Table 1. 14. A bar graph showing the comparisons of the heartbeat of Daphnia sp. with treatments (addition of caffeine solution) and without treatments is plotted 15. A line graph of the heartbeat of Daphnia sp. per minute against the concentration of caffeine solution is drawn

RESULT Concentration of caffeine solution (%) 0.0 (control) 0.1 0.2 0.3 0.4 0.5 The heart rate of Daphnia sp. (beats per minute) Reading 1 328 376 392 396 376 372 Reading 2 336 384 400 412 384 368 Average 332 380 396 404 380 370

Bar Graph of Heart Rate of Daphnia sp. (beats per minute) against The Concentration of Caffeine Solution (%)
Heart rate of Daphnia sp. (beats per minute) 450

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0 0 0.1 0.2 0.3 0.4 0.5 Concentration of caffeine solution (%)

Graph of heart rate of Daphnia sp. (beats per minute) against the concentration of caffeine solution (%)
Heart Rate Of Daphnia sp. (beats per minute) 450

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0 0 0.1 0.2 0.3 0.4 0.5 0.6 Concentration of Caffeine solution (%)

Increment of Heart Rate of Daphnia against the Concentration Of Caffeine Solution (%)

Increment of Heart Rate of Daphnia 80

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0 0 0.1 0.2 0.3 0.4 0.5 0.6

Concentration Of Caffeine Solution (%)

DISCUSSION
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The aim of this experiment is to investigate the effects of concentration of caffeine on the heart rate of living organisms. In this case, to investigate the effect of concentration of caffeine on the heart rate of Daphnia sp. or more commonly known as water fleas. The hypothesis is that the greater the concentration of caffeine solution the higher the heart rate of Daphnia sp.. While conducting this experiment, we can also equip ourselves with laboratory skills and investigative skills. The independent variable of this experiment is the concentration of caffeine solution. This variable is controlled by measuring different amounts of caffeine powder (coffee powder) that is 0.1g, 0.2g, 0.3g, 0.4g and 0.5g using an electronic balance and dissolving the caffeine powder in exactly 100ml of distilled water. By using this method, we obtain caffeine solutions of different known concentrations that is 0.1%, 0.2%, 0.3%, 0.4% and 0.5%. Distilled water is used instead of tap water because distilled water is pure and does not contain any other organisms that may disturb our view of the Daphnia sp.. The concentration of caffeine solution is set from 0.1% to 0.5% only because in this concentration the Daphnia sp. can still stay alive and carry out its metabolic functions as usual thus making the experiment possible. If caffeine solution of concentration higher than 1.0% is used, the Daphnia sp. will automatically die when exposed to this solution, making it impossible to carry out the experiment. The dependent variable in this experiment is the heart rate of Daphnia sp.. The technique used to measure this is by observing the heart beats of the Daphnia sp. which can be seen clearly when viewed under a microscope. The heart rate of Daphnia sp. when in its natural habitat that is in the pond water without any treatments is set as a control so that comparisons between the heart rate in different caffeine concentration can be made. The heart beats of the Daphnia sp. can sometimes be hard to observe. Another method that can be used to count the heart beat is by observing the leg beat of Daphnia sp. which is proportional to its heart beat. To make observations and counting of the heart beat easier, a scientific calculator is used. Another method involves tapping a pencil on a piece of paper and counting the dots made afterwards. The best way to count the heartbeat is actually by capturing a video of the Daphnia sp. in the 15 seconds time frame and replays it in slow motion to count the heart beats. However because of limited resources, the calculator method is chosen. The constant variables in this experiment include temperature and pH of the environment of Daphnia sp., time to record the heartbeat of Daphnia sp. and the type of
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Daphnia sp. used. The temperature and pH of the environment of Daphnia sp. is controlled by carrying out the experiment in an air conditioned laboratory where the temperature is set to 25oplacing the Daphnia sp. in a culture containing pond water from its natural habitat. Other than that, distilled water is used to make the caffeine solutions to ensure a stable pH where the Daphnia sp. can survive in. the time to record the heart beats of the Daphnia sp. is set to 15 seconds by using a stopwatch. The counted heart beats in 15 seconds is then multiplied by 4 to obtain the number of heart beats per minute. The same type of Daphnia sp. from the same species and breed is used for every repetitions of the experiment.

FURTHER STUDIES

SAFETY PRECAUTION Throughout the experiment, some safety precautions are exercised to ensure the accuracy and reliability of the results. 1. Lab coats and closed toe shoes must be worn at all times inside the laboratory when carrying out the experiment to protect ourselves from any injuries caused by broken apparatus. 2. Eye protections must be worn to protect ourselves from any toxic chemicals or any microorganisms that is found inside the pond water of the Daphnia sp. culture. 3. The experiment must be done in an air conditioned room which temperature is set to below 30oC. This is because Daphnia sp. are cold climate creatures and may die from high temperatures. 4. When carrying out the experiment, students must treat the Daphnia sp. very carefully as the number of Daphnia sp. available in the laboratory is very limited. Therefore students need to be extra careful when handling the Daphnia sp. to avoid them from dying. 5. When observing the Daphnia sp. under microscope, ensure that the light used is not so bright. This is to prevent the Daphnia sp. from dying from overheating due to the exposure to bright light.

6. When placing the Daphnia sp. on the cavity slide, make sure that there is only a small amount of water on it. This is to prevent the Daphnia sp. from moving around on the cavity slide which makes it hard to observe under the microscope. 7. In contrast, ensure that there is enough water on the cavity slide to ensure that the Daphnia sp. can survive. Too little water causes the Daphnia sp. to suffocate and die. 8. Filter paper should be used in this experiment instead of muslin cloth as suggested by the manual. This is because the muslin cloth is too absorbent and may remove the Daphnia sp. from the glass side. In addition, muslin cloth has fibres that may stain the cavity slide and make observations of Daphnia sp. harder. 9. The experiment for each concentration must be repeated at least twice to obtain the average count of heart beat. This is to increase the validity and reliability of the experiments 10. A new Daphnia sp. should be used for every different concentration of caffeine solution. This is to avoid the Daphnia sp. from developing tolerance towards caffeine which cab affect the results greatly.

LIMITATIONS There is unavoidable existence of limitations that could affect the outcomes of the experiments. They are 1. When the Daphnia sp. is placed on the microscope stage and exposed to the microscope light, it can cause the Daphnia sp. to be overheated. This can increase the heart rate of Daphnia sp. which affects the validity of the results. This can also cause the Daphnia sp. to die from overheating. 2. The heartbeat of Daphnia sp. when immersed in caffeine solution of 0.3% to 0.5% is very fast. Missing a few heartbeats when counting is inevitable. Therefore the results obtained may or may not be accurate to the actual number of heartbeats of the Daphnia sp.. 3. The use of scientific calculators may also affect the number of heartbeats counted. Because the tapping of the calculator is very fast because of the fast Daphnia sp. heartbeat, the number of tapping and the number that is shown on the calculator screen may not be the same thus affects the results of the experiment.
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4. The same Daphnia sp. is used by two or more groups therefore it may be injured or hurt while being transferred from one container to the other. Therefore the heartbeat rate may change and affects the results. 5. Long exposure of the Daphnia sp. to caffeine solution causes the Daphnia sp. to develop tolerance against the caffeine solution where the heart rate does not increase even when the concentration of caffeine solution is increased. Therefore the results is not accurate or obey the theory/hypothesis anymore. 6. Since the Daphnia sp. is not observed under its natural habitat, its stress level may increase which causes a fluctuation in its heart rate. Therefore the counting of the heartbeat must be done as soon as the Daphnia sp. is ready to be observed.

CONCLUSION REFERENCE 1. Edexcel AS Biology, Students book, Pearson company, 2008 2. Edexcel Biology for AS, CJ Clegg, Hodder Education, 2008 3. Text PRE U Biology Volume 1, Lee Ching, PEARSON Longman, 2008 4. International Edition Biology Seventh Edition, Campbell, Reece, Pearson Benjamin Cummings, 2005 5. Advanced Biology Principles & Applications Study Guide, CJ Clegg with DG Mackean, PH Openshaw and RC Reynolds, John Murray (Publishers) Ltd, 1996 6. Carpenter, SR, & Kitchell, JF (1993) The Trophic Cascade In Lakes. Cambridge University Press, London, England. 7. Davidson College Department of Biology (1999)

http://www.bio.davidson.edu/index.html. 8. Rith, J (1988) Plant succession on abandoned railways in rural New York State. Proceedings of the 73rd Annual Meeting of the Ecological Society of America, Davis, CA. 9. www.sciencebuddies.org/mentoring/project_ideas/Pharm_p009.shtml 10. http://en.wikipedia.org/wiki/Daphnia sp. 11. http://en.wikipedia.org/wiki/Caffeine 12. http://www.caudata.org/Daphnia sp./

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