The effect of caffeine on heartbeat of Daphnia

Introduction
In this experiment, I am going to investigate the effect of caffeine on
heartbeat of daphnia. The aim is to see whether the caffeine will affect the
heart rate of the daphnia or not. Also I will be doing a blind study as it is
hard to record the number of heart beats per minute. I will also check the
reliability of the experiment in certain ways, and also check for
improvements that can be done.
Hypothesis
As the concentration of caffeine increases in the solution containing
daphnia, the daphnias heart rate increases.
Background Biological Information
Daphnia is a genus of planktonic crustaceans. It is usually 1-5mm long and
lives under water in various water environments e.g. acidic swamps, fresh
water lakes, ponds and rivers. They are also called water fleas because of
their saltatory swimming style which resembles of fleas. Daphnias body is
divided into segments. It has a head fused into the body and is bent down
towards it, with a notch which separates the head from the body. Most of
the species have their body covered by carapace with a ventral gap where
the six pairs of legs lies on. It also has got compound eyes, the antennae
and a pair of setae. In most species of daphnia, the skin is translucent, and
sometimes transparent.
The daphnias average heart rate is normally 180 bpm. When it gets a dose of
caffeine, its heart rate will increase because of the stimulants that are triggered by
the coffein.

Caffeine is medically known as trimethylzanthine and its chemical formula

is C8H10N4O2. When in pure form, caffeine is a white crystalline powder
that tastes very bitter. The most common way of acquiring pure caffeine is
the process of decaffeinating coffee and tea. Caffeine is useful as a cardiac
stimulant and also as a mild diuretic. Cardiac stimulants increase the heart
rate, and diuretics increase urine production. Caffeine is also used to
provide a boost of energy or a feeling of heightened alertness.
Independent Variable:
The independent variable is the variable which I will be changing, and in
this case it will be the concentration of caffeine in the solution that I will
drop on the daphnia. I have chosen a range of 6 values which are 0% 0.1%,
0.2%, 0.3%, 0.4% and 0.5% of caffeine in water.
Dependant Variable
The dependant variable is the variable that depends on the values that I
choose, which in this case will be the results that I produce from the
experiment, and it will be the number of heartbeats in one minute.

Variables to control
In this experiment, I will have to set some variables that I should keep
constant to get reliable results:
- Temperature, as I need to maintain the same temperature in every
trial trough the experiment, and it will affect my results if it changes.
To do this, I will make sure that I stay at the same room doing this
experiment through every trial, at normal room temperature. Also the
light bulb that I will use in this experiment with the microscope heats
up and increases the temperature of the surroundings including the
daphnia, so
- Amount of caffeine solution I will need to add to the daphnia when its
under the microscope. It should be two drops from the pipette. This
could affect the results because of the spreading of the caffeine
molecules throughout the water solution.
- The size of the daphnia used, as it needs to be the same throughout
the experiment. It affects the results because the surface area to
volume ratio could differ between the daphnias which leads to
differentiation in the diffusion of the caffeine solution and therefore
leading to unpredicted results.
Apparatus
In this experiment, I will need:
Culture of daphnia
Cavity slides
Pipettes
Distilled water or pond water
Normal water
Caffeine solution of concentrations 0.1%, 0.2%, 0.3%, 0.4% and 0.5%
Cotton wool
Standard glassware (beakers, measuring cylinders etc.)
Stopclock
Paper towels
Microscope and lamp
Method
1.
Use a pipette to transfer the daphnia to a cavity slide containing
pond water of known volume.
2.
Add some cotton fibres to the cavity slide and move them in such a
way that the daphnia cant move.
3.
Use a paper and a pencil to put a dot on the paper every time the
heart beats.
4.
One student will use a stopwatch to count 60 seconds which is the
time during which the measurement of the heart rate will take
place.

5.
6.
7.
8.

Turn the microscope lamp on and locate the heart of the daphnia
through the microscope.
Measure the beat rate by putting the dots on the paper every time
the heart beats until the stop watch reaches 1 minute.
Repeat steps 1-6 at least 5 times and get an average. Then count
the heart beats per minute.
Clean the cavity slide using a tissue and make sure there is no
pond water left.

Control
The control I have chosen for this experiment is the 0% concentration that
easily gives us an estimate of the normal heart rate of the daphnia. From
this control, I will be able to compare the other results that I can get, and I
will be able to manipulate data.

Risk Assessment
In this experiment there are some risks that I should be conscious of when
carrying out the experiment which can be fatal to both me and the daphnia:
- When I carry out the experiment I should be cautious not to touch the
lamp with wet hands since it uses electricity, and it also heats up and
it could burn my skin.
- The pond water should not be inhaled as it might contain bacteria and
viruses that cause diseases. Also it might affect someone else who is
sensitive to some of the particles found in the pond water, so I will be
careful when moving around with the pond water.
- As for the daphnia, there is a chance for them to die when they are
exposed to light from the bulb which transfers heat which they arent
used to getting exposed into, so I will switch off the bulb between
measurements and calculations.
- When the daphnia are given caffeine of a too high concentration their
heart may beat so fast that it will cause their death, so I will be
choosing the largest daphnia from the culture of daphnia, and make
sure to choose the right concentration of water.
- Furthermore, there is the risk of the daphnia to dry up when not
exposed into water, which may kill them in the middle of the
experiment, so I should make sure that they get enough water or
caffeine-containing water.

Table of Results

Concentration of caffeine in the tube

0 beat per
0.1
0.2
0.3
0.4
Heart
99 (bpm)126
229
247
257
minute
76
121
241
118
120
145
229
111
136
233
287
55
88
62
75
87
124
142
238
257
267
182
276
196
206
176
181
224
Average
110
136
226
231
254

(%)
0.5
289
283
276
52
291
180
264

Calculations
I used the results of the trials to calculate an average of them. The formula I
used was:
Average= Sum of trial results
Number of sums
Trends and Patterns
The graph shows that generally, the heart rate per minute increases as the
concentration of the caffeine increases. There were outliers that didnt seem
to fit into the graph, so I took them out of the range bars. However, the
range bars were still big in the first two and the last two.

Conclusions
So far I have done the experiment and I found out that although there was
an increase in the heartbeat with the increasing of the concentration of the
caffeine in the water, the range bars were big and they didnt quite support
the hypothesis. However, if we look at the averages of the trials, we will
realise that there is an increase in heartbeats, which supports the
hypothesis and what was included in the biological knowledge.
Explanation of Results
The results gained from the experiment and their trend and patterns on the
graph could be explained somehow. The outliers identified could have
happened because of human error as it couldnt be easy to catch up with
the fast heart rate of the daphnia and record it all in one minute. Also the
daphnia may have stood still in a small amount of water for a long time

leaving it stressed which could have affected its heart rate. Also it may have
been exposed to too much light and heat from the bulb which affected its
heart rate.
Reliability of conclusion
This experiment could be repeatable, but not that reliable because of the
limitations of the method. This could be explained as results support the
hypothesis, but some of the patterns found dont support the hypothesis,
which made it unreliable.
Variability of Data
This data that I have collected seems to have some variability of over 10%,
which I identified as outliers, and some of these that I have included in the
range bars may as well have gone beyond that point. But the average seems
to be well enough as it followed my hypothesis and background information.
Limitations of the method
This method seems to be hard to go through as it does not give accurate
readings, and it depends solely on whether the person using this method is
able to catch up with the heart beats of the daphnia and record them all.
This was limited because this method doesnt require a recorder of the
heartbeats of the daphnia, which could have solved the whole problem.
However, I was happy with the repetitions of the experiment as I could do at
least 5 repetitions per concentration, which lead me to collect much data to
rely on.
References
http://en.wikipedia.org/wiki/Daphnia
http://en.wikipedia.org/wiki/Caffeine
http://www.education.com/science-fair/article/effect-of-caffeine-onheartbeat/
http://web.as.uky.edu/Biology/faculty/cooper/ABLE/Daphnia_Concentration_
Paper.pdf