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What are Daphnia, and why are

they used?

How does caffeine

affect the heart?

- Daphnia, also commonly known as water fleas, are

- Caffeine increases the activity of

small crustaceans that live in lakes, ponds, streams or most fresh water.

- They serve as an important part of the food chain, as

food to fresh water fish and other organisms living in
these fresh water conditions.

They are 1-5 millimetres long with a translucent

exoskeleton, that is split up into segments. Their
heart rate is approximately 180 bpm.
Their heart is located behind the head, at the top of
their back, which means it is easy to identify. The
heart beat can be clearly observed through a
telescope and so Daphnia are used to observe the
effect of caffeine.

- Their small bodies mean that caffeine quickly

diffuses into their blood stream, causing a very quick
change in heart rate.

- Daphnia can be easily raised and kept in an

aquarium in a classroom environment, with a

the heart.

- The sinoatrial node (controls the

beating of the heart) has an
increase in electric activity,
which causes the heart to beat at
a faster pace.

- There is an increased contraction

and relaxation rate which results
in more activity in the ventricles
and atria, as well as an increase
in the volume in blood pumped
during a specific length of time.


the heart rate

experiment using Daphnia

Ethical Issues & Considerations:

Daphnia are very small with a less sophisticated nervous
system and are small meaning their are easy to handle
carefully, which means that they are going to be less
susceptible to pain, as opposed to mammals or bigger
animals. They are easily and quickly accumulated in a
classroom environment, meaning they do not need to be
removed from their natural habit, such as a river, for the
experiment to take place. The heart beat of the Daphnia can
be recorded without the use of any invasive technique or
equipment, unlike those that would be required for a
mammal. Carrying out this process requires you to handle
the Daphnia very carefully. Unfortunately, Daphnia are not
protected by any law stating., therefore the vulnerability
for the invertebrate is not always considered. However, it is
important to be mindful of these animals and respect that
they are conscious animals who, undergoing this
experiment, would be significantly scared. These small
invertebrates are being taken from their pond water
environment and transferred in a narrow pipette, which
would be frightening, and so the transportation of the
Daphnia should be quick but careful.
The lamp from the microscope produces heat, which can
warm up the water that the Daphnia are surrounded by on
the slide which, as well as increasing their heart rate, could
harm the crustaceans. Therefore, the time that they are left
under the microscope must be monitored and kept short.
Daphnias natural environment is that of pond or river water
as it is of the correct pH and minerals. Tap water and the
caffeine in the solutions are an unnatural environment for
these invertebrates and could potentially harm them. The
time they spend in these solutions must also be kept brief to
ensure minimal harm comes to the Daphnia.

Carrying out the experiment

The equipment needed for this experiment includes:
Glass slides; Microscope; Caffeinated energy drink; Daphnia in pond
water; cotton wool; pipettes; timer; beakers;
Gather a sample of Daphnia in a beaker with enough of their pond water
to stay in. Take 2 beakers, in the first, place a quantity of purely
caffeinated energy drink, the next with a 1:1 ratio of energy drink and tap
water. On a glass slide, create a barrier with a small amount of cotton
wool, so that the invertebrates stay still. Using the pipettes, pick up one of
the largest Daphnias from the beaker of pond water and place in onto the
slide within the cotton wool barrier. Position the slide on the microscope
and locate the heart, so that you are able to clearly see the heart beats.
With a pen and paper, dot the paper each time the heart beats for a
duration of 30 seconds. Immediately remove the Daphnia from the slide
and place back into the pond water. Count up the dots and times the
number by 2 in order to get the bpm. Take the pipette and carefully
remove a different larger Daphnia. Place it into the 50% caffeine solution,
and time for 30 seconds. Once the time is up, remove the Daphnia and
place onto a glass slide and then repeat the method for recording the
heart rates. Repeat this method with the 100% caffeine solution.
Independent variable: Concentration of caffeine
Dependent variable: Heart rate of the Daphnia
Control variables: Time spent counting the heart rate; form of caffeine
used; time Daphnia spend in caffeine solution; age of daphnia; size of
Daphnia; pre-treatment of Daphnia; temperature of surroundings; oxygen
concentration in water; (every aspect besides independent variable
should be controlled)
To ensure VALID results, each control variable must be monitored and