Biology Practical Report Name:

Intake: April 2012

Student I.D: 100123

Title: Investigation of the effect of caffeine on heart rate.

Date of Experiment: 25/7/2012

Date of Submission of Report: 1/8/2012

Topic: To investigate the effect of caffeine on heart rate. Introduction: Caffeine is an odourless and slightly bitter alkaloid. It is most commonly consumed by humans mostly by drinking coffee or tea, as well as from various foods and drinks containing products derived from the kola nut. Early experiments showed that low concentrations of caffeine may produce small decreases in heart rate in human, whereas higher concentrations may make the heart beat abnormally fast. In the brain it constricts the cerebral blood vessels. If one is used to drinking several cups of coffee a day but quit drinking later, those blood vessels will dilate, the dosage perhaps is enough to give one a major headache. It is one of the best known withdrawal symptoms.

When caffeine is used in moderation, it acts as a mild stimulant to the nervous system, blocking the neurotransmitter adenosine. This results in a feeling of well-being and alertness. Excessive intake of caffeine can lead to restlessness, insomnia, and heart irregularities. The effects of caffeine vary from person to person, as people excrete it at different rates and each individual has a different genetic structure. Physical dependence and unpleasant symptoms upon withdrawal (headache, fatigue and depression) are common in regular caffeine users. A study of effect of caffeine in human heart rate will be very valuable. However, there are many difficulties in conducting an experiment that involves human as guinea pig due to the low birth rate, long life cycle and the violation against human rights. Thus, an alternative is to use a model organism to represent the human. The results of the experiment can then be used as a reference to the effect of caffeine on the heart rate of human. Daphnia are selected as the model as they can be easily handled in many ways in the investigation. Between 0.2 to 5mm in length, Daphnia are small, planktonic crustaceans which are also commonly known as freshwater cladocerans, often classified with other tiny crustaceans as microcrustaceans. Cladocerans are commonly known as water fleas. This is not only due to their salutatory swimming style, but also because of their resemblance to real fleas, though real fleas are insects and shares only an extremely distant common ancestry with Daphnia, since both crustaceans and insects are arthropods. All species of Daphnia occur in different strains - sometimes the same species can look completely different, both in terms of size and shape, depending on its origin, and environmental factors at that location. The life span of Daphnia, from the release of the egg into the brood chamber until the death of the adult, is highly variable depending on the species and environmental conditions. Usually, the lifespan of a Daphnia does not exceed one year and it is largely related to the surrounding temperature. It is discovered that the life span increases as temperature decreases, due to lowered metabolic activity. Daphnia reproduce through parthenogenicity, which is the ability to self-replicate without fertilisation of any form, during the late spring, summer and early autumn, depending on temperature, food availability and presence of waste products of their metabolism. Once thought of as an animal of polluted waters, Daphnia have been proven to be very sensitive to poor water conditions and a number of research and industrial groups have used these organisms as test subjects to evaluate the water quality. For example, they are very sensitive to halide concentration, like the chloride or fluoride in tap water, which are extremely toxic to Daphnia. They are also sensitive to metal ion concentration, like sodium, potassium, magnesium and calcium, which in increased concentrations can cause immobility and death, and Daphnia are extremely sensitive to copper, zinc and most dissolved toxins. Often, they are used to monitor water quality so that only safe water is released into the environment by industry and water treatment plants. Similar to many animals, Daphnia are prone to alcohol intoxication, which makes them excellent subjects for studying the effects of the depressant on the nervous system due to the translucent exoskeleton, and the visibly altered heart rate. They are tolerant of being observed live under a cover slip and appear to suffer no harm when returned to open water. In this experiment, it is intended to study the effect of different concentration of caffeine solutions on the heart rate of Daphnia.

Method: 1. Two cavity slides are prepared. The first cavity slide is for the tap water and another one is for the Daphnia. 2. Caffeine solutions of concentrations of 0.5%, 1.0% and distilled water are prepared. 3. One Daphnia is selected and scooped cautiously from the container using a Petri dish along with moderate amount of water. 4. The Daphnia is transferred from the Petri dish onto a cavity slide, by using a dropper, along with a small amount of pond water.

5. The surrounding of the cavity slid is dapped dry in order to restrict the movement of the Daphnia when placed under the microscope to be observed. However, there are still drops of distilled water present in the cavity slide to prevent the Daphnia from dying. 6. Cavity slide Daphnia is placed on top of cavity slide tap water and the microscope is switched on. For the 1st minute, the Daphnia is allowed to adapt to its surrounding. After 1 minute, the stopwatch is started. For the next 30 seconds, as the heart of the Daphnia beats, a dot is tapped on a piece of paper. Once the 30 seconds is up, the Daphnia is allowed to rest for another 30 seconds and the tapping on the paper also stops. The same procedure is repeated for 3 minutes. 7. If the Daphnia doesnt die, steps 1 to 6 are repeated using the 0.5% and 1.0% caffeine solution.

Results: Mean of Heart Rate/30 sec Readings 1 Solution Distilled water 0.5% caffeine 1.0 % caffeine 55 136 159 56 132 130 57 132 160 56 133.33 149.67 2 3 Average Mean of heartbeat per minute 112 266.67 299.33

Discussions: 1. The result of the experiment is correct because the rate of heartbeat of Daphnia increases in 0.5 % caffeine solution and 1% caffeine solution. 2. As the concentration of the caffeine solution increases from 0.5% (control) to 1.0%, the graph shows an upward trend. 3. As the concentration of caffeine increases, heart rate of Daphnia increases as caffeine concentration up to 0.5% till 1.0% because caffeine is a stimulant which raises the heart rate of animals. Assumption that a similar effect would result in human can be made. 4. Excess water is removed by using tissue paper from the cavity slide as too much solution will result in vigorous movement of Daphnia. Counting of heart beat or beatings of the legs would then be more difficult. 5. There are a few unavoidable limitations in this experiment which may affect the accuracy of the results. The following are the limitations and their respective improvements: Temperature of the environment of the Daphnia is one of the factors that affect the heart rate of it. Prolong exposure of the Daphnia to the microscope light will overheat the Daphnia. To prevent the Daphnia from overheating, the microscope light should be turned off between observations and a cavity slide with tap water is placed beneath the cavity slide containing the Daphnia is used. The Daphnia may be hurt and injured while it was being transferred from one container to the other. This may affect the experimental results. The only step can be taken is to handle the Daphnia carefully to avoid injuring it. Since the Daphnia is not observed under its natural habitat, its stressed level in the experiment may be higher than usual. This might cause fluctuations in its heart rate or an undesired change

in the heart rate. The only thing to be done is to count the heart beat or beating of legs of the Daphnia as soon as it is ready to be observed. It should be understood that any experiment involving living organism always rises up bioethical issues. While performing experiment involving living organism can bring greater advances in science, it should not be forgotten that the well-being of the living organisms involved in the experiment should be taken care of, so that they do not suffer for the experiment or at least their sufferings are minimized. Therefore, the Daphnia being used in the experiment ought to be treated with great care to avoid hurting or even killing them. If possible, the Daphnia should be returned back to their natural habitat after the experiment so that they can return to their normal lives. This step also avoids any disruption in the food chain which may affect the ecosystem.

Conclusion: As a result, as the concentration of the caffeine solution increases, the heart rate of Daphnia increases.