Activity 1.23: Does Caffeine affect heart rate?

What treatments are you investigating in this experiment?

Control (distilled water) Vs 0.2% caffeine solution

Prior to examining the Daphnia under the microscope you should allow time (e.g. 35 minutes) for the Daphnia to acclimatise to the solution in which you have placed
them.
1. Place a few strands of cotton wool on a cavity slide to restrict the movement of
the Daphnia. Using a pipette transfer one large Daphnia to the cavity slide,
keeping plenty of water surrounding the flea. Do NOT use a cover slip! Using the
microscope on the lowest magnification, focus on the Daphnias heart, which can
be see through its translucent body.
2. Use a pencil and paper (or whiteboard and pen) and a stop clock to record the
number of heartbeats in 30 seconds.
3. Repeat the process a minimum of two more times and calculate the mean.
Remember that if you identify any outliers as you are doing your repeats you
should discount the outlier and repeat again.
4. Repeat steps 1-3 for a different treatment (i.e. control vs caffeine OR five
different concentrations of caffeine) using a different Daphnia. Work in pairs
using the blind technique.
Topic One Core Practical: Effect of caffeine on Daphnia
Hypothesis
H: Daphnia heart rates will be higher in the 0.2% caffeine solution compared with the control
as the caffeine acts as a stimulant, causing stimulatory neurotransmitters to be released.
NH: Daphnia heart rates will be unaffected by the higher concentration of caffeine in the 0.2%
solution

Variables
Independent Variable Caffeine concentration
Dependent Variable Daphnia heart rate
Control Variables
-

Temperature
Volume of solution
Stress of daphnia
Time of acclimatisation

Control Variable
Temperature

Volume of solution

Stress of daphnia

Time of
acclimatisation

How it will be controlled?

Temperature will be controlled
by carrying out each test in
the same room and keeping
the door shut to reduce risk of
temperature change between
experiments.
Volume of caffeine solution will
be controlled by taking each
beaker of solution from the
same 2.0% mix so that there is
no variation between
experiments.
Stress of daphnia will be
controlled by only using nonimpregnated daphnia that are
all the same size. Their time
under the microscope must
also be the same throughout
the tests so as to keep stress
levels caused by the light of
the microscope the same.
Time of acclimatisation will be
controlled by timing the
amount of time each daphnia
spends acclimatising to the
solution/control; each daphnia
will spend 5 minutes in their
respective solution/control.

Why are we using Daphnia in this experiment?

Why must it be controlled?

Amount of caffeine in the daphnias

blood will affect its heart rate, meaning
that variation of caffeine solutions will
skew results.

Increased stress levels will increase the

heart rate of the daphnia, meaning
that a variation in stress of each
daphnia could skew results.

Amount of caffeine in the daphnias

blood will affect its heart rate and the
time taken to acclimatise to the
caffeine solution will affect the amount
f caffeine in the daphnias blood. This
means that a variation in
acclimatisation time will skew results.

Ethics
When using living organisms in experiments it is important to be aware of the ethical implications. For
example, what precautions did you take to ensure their safety? Is it appropriate to use the animals in
scientific research? Etc.
This webpage should help your understanding of ethics. Have a quick look to check your
understanding: http://www.bbc.co.uk/ethics/introduction/intro_1.shtml

1. What is ethics?

2. Think of at least three ethical arguments which are either for or against the use of
Daphnia in this practical.

Argument

Ethical Point

Further explanation this point/argument

3. Ethical Question: What is the utilitarian argument for using living organisms in scientific
experiments? Do you think this ethical argument supports the use of Daphnia in this particular
experiment?

Now complete the following and add to your core practical report
Results Table (including repeat columns, mean and standard deviation).
Graph of results (Including error bars representing standard deviation)
Discussion (Refer to the Practical Support 1 document Planning and writing up an
experiment which is available under the tab skills support on SNABonline.com to help you
complete this section)
Evaluation (Discuss the possible limitations of this technique and how they could be
overcome in this section, not in the discussion)
References (Any references e.g. journals etc., cited in your discussion should be
referenced in FULL in this section. You are expected to add such research in to your report
particularly as it is a skill required for your AS coursework. There is a document dedicated
to referencing on the Moodle AS Biology page accessible via the SMRT website).
Super Sauce Question (This should be added in between the evaluation and the references
section)
A)
Caffeine affects neurotransmitter levels in the brain. Daphnia have no brain, so how did the
caffeine produce its effect?
B)
Can we extrapolate our findings on Daphnia directly to humans? Give a reason for your
answer.