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4. Using the stopwatch, count the heart beats of the Daphnia by putting paper to
pen every time you see the heart of the Daphnia beat. This is useful as it ensures
minimal human error due to the fact that you arent clicking anything which could
malfunction. Make sure that you are dotting a different part of the paper at every time.
This will be a control to be compared with the other results as it they are all affected by
caffeine. (Make sure the same person does it everytime to reduce Random errors)
5. Place a couple of Daphnias in another small container with pond water and place
0.1% of Caffeine (or the other concentrations of caffeine) in that measuring cylinder.
Keep the Daphnia in it for 3 minutes.
6. Then repeat steps 1-5 for every concentration of Daphnia.
7. Repeat the experiments 3 times with different organisms of Daphnia for the
different concentrations.
8. Obtain the results and record the average.
Safety Precautions:
The Daphnias must be handled carefully and not roughly. Handle all the experiments carefully
and if any chemical touches skin, ensure that the area exposed is immediately washed. Wear
safety goggles to prevent chemicals from splashing into the eye. Make sure hair is tied back at
all times and it does not touch any of the chemicals. When handling liquid with temperatures,
take precaution needed (such as using tongs) to prevent scalding of hands. Also, when handling
liquid, ensure that the liquid doesnt spill anywhere that might cause someone to trip and fall.
Hygienic precautions are also very necessary as a culture is used. Always make sure that the
microscope is carried by 2 hands, to ensure that it doesnt fall and break.
Graph:
Analysis/Discussion:
The results from our experiment shows us that the caffeine concentration has various effects on
the Daphnias. At the start, there is a 3.5 % decrease of the mean beats/minute of the Daphnias
from the Caffeine concentrations of 0.1 % to 0.2 % which suggests that caffeine is a depressant
rather than a stimulant. The trend then increases from 0.2% to 0.4% with there being an 11.8%
increase of the Daphnias heart rate from 0.2 % to 0.4 %. However, after that, there is a
decrease in the heart rate, from 186 to 170, which is an 8.6 % decrease in the mean
beats/minute of the Daphnias. The results of the control experiments, however, were much
more varied. There is a 38.6% difference between the fastest heart rate of the Daphnias and the
slowest heart rate. However, the results of the Daphnia with the mean of 194 beats/minute can
be considered an anomaly as it is extremely high and does not match the other results and has
a 14 beat/minute difference with the average heart beat of a Daphnia, which can be considered
to be quite high. However, the slowest heart rate of the Daphnias that were under the control
condition, with only 140 beats/minute can also be considered an anomaly as it is too small and
is too low compared with the other results, with there being a 22 beats/minute difference with
the second lowest result and is also 20 beats/minute off the average heartbeat of a Daphnia,
which is at 180 beats/minute.
The result of this experiment also does not match our hypothesis in the sense that the average
heart beat of the Daphnia is at 180 beats/minute but almost every result, except 1, is lower than
the average heart rate, despite the concentration of the caffeine increasing. The only result that
matches our hypothesis could then be regarded as an anomaly as it is the only one that has
surpassed the average heart rate. However, based on the control in this experiment (as these
Daphnias could have been obtained in a different pond or a different region to the Daphnias that
were tested to prove the average heart rate and those Daphnias could have a higher tolerance
to caffeine), which then suggests that the average heart rate is too high for the Daphnias that
were used in these experiments.
The science behind this experiment should be that the caffeine acts as a stimulant to the
Daphnias and increase their heart rate, thus making them more active. As their heart rate
increases, their heart should pump faster, which should be observable under the light
microscope. The higher the concentration of caffeine, the faster the heart rate as there is more
of the caffeine running in the bloods of the Daphnia.
Conclusion:
The results from this experiment do not support our data by much due to the Daphnias and that
they can be affected easily by temperature (which could easily change due to the surroundings
and the fact that this experiment was not done in a professional laboratory. The temperature
could also change due to the light from the microscope, which increases the temperature of the
water). The anomalies, illustrated by the control (194 beats/min), could also be due to the high
stress levels as the Daphnias are not in their natural environment. However, if an average of all
the control is taken, it can be concluded that the hear rate of the Daphnia does increase, as a
general trend, as the concentration of caffeine increases.
Evaluation:
Due to time constraints, we couldnt select the Daphnia that was the biggest, which would have
helped us as we measured the heart rate as the heart would have been bigger or it would have
been more obvious, which would have been more obvious. Besides that, due to the fact that all
the Daphnias were of different size, some heart rates were easier to measure than others.
However, larger daphnia will have a slower response to change in caffeine than a smaller
daphnia as the surface area to volume ratio is smaller due to the longer diffusion distance. It
also would have been better if we purchased or kept the age of the Daphnia the same (as far as
possible), by keeping them from when they have been hatched. This is because the various
caffeine concentrations could have a different effect as the age of some Daphnias increase as
(with humans), the arteries harden and lose its elasticity, and therefore it is harder for the
arteries to accommodate to the higher blood pressure surging through. This then reduces the
validity of our results as the Daphnias are of different age and react to the caffeine
concentrations differently and we do not know the age of the Daphnia sample that we
experimented on.
Besides that, the validity of the experiment could have been improved on as the temperature of
the solutions are kept the same. This is because a high temperature could affect the Daphnia by
making it more active (the enzymes would be closer to its optimum temperature, thus working
faster and making the reactions in the Daphnia to happen at a quicker rate), thus increasing its
heart rate. If the temperature gets too high, then the Daphnias could potentially die due to the
high temperature and the inability for the body of the Daphnia to accommodate to it.
We controlled the reliability of our results by ensuring that the confounding variables are kept
the same as far as possible. This could include the temperature of the water, the pond that the
Daphnias were obtained from and this could also include the type of Organism.
Besides that, to improve the accuracy of our results, we could have also made sure that the
Daphnias were kept at 10 degree celcius before experimenting on them as this is when the
heart rate of the Daphnias are at their slowest rate (this also ensures that the heart rate of the
Daphnias before they are mixed with the caffeine is as similar as possible). To improve the
accuracy of our results, we also need to make sure that the used Daphnias are not returned
back into the container with the original Daphnias as this could cause a mix up and we could
accidentally re use the same Daphnia for another experiment of a different caffeine
concentration. To also improve on the reliability of our results, the volume of caffeine placed with
the Daphnia needs to be kept as similar as possible to ensure that there is no overexposure of
the caffeine to certain Daphnia molecules, which could affect the heart rate of the Daphnia by
increasing it, thus decreasing the reliability of our results as the result wouldnt be as accurate
as possible. We also have to make sure that we do not keep the Daphnias under the
microscope for too long as if left too long under light microscope, temperature increases due to
heat of lamp and causes increase in heart rate.
To improve the precision of our results, we had used a stopwatch instead of a clock to ensure
that the time is read to the last second to make sure that it is as similar as possible for every
repeat. Also make sure to turn off the light on the microscope when not observing daphnia
during period of acclimatization to prevent overheating of daphnia. Start stop clocks at the same
time to ensure that the result is as precise and as accurate as possible. We could also improve
the precision of our results by using a clicker instead of just putting pen to paper and this could
minimize human error.
In addition to that, since the Daphnia is not observed under its natural habitat, its stress 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. Weak daphnia or pregnant
daphnia may be more easily stressed than normal healthy daphnia.
Besides this, standard deviation shows how close the values are to the mean (how spread the
data is). Our data lacks accuracy as the values are spread and far from the mean.
Further Experiments:
To further analyse the effect of a drug, like caffeine, on the heart rate of a living organism, like
the daphnia, these are some changes in the experiment that could have been done:
Use another species of daphnia, to see if the results are the same. This could be
done on brine shrimps or other organisms with a similar circulatory system.
Use different concentrations of caffeine, such as 0.3% and 0.6%. This would
validate our results even more as it proves our results to be more reliable.
The results of the experiment would be more accurate and valid if a blind study
is done. This is because the experimenter who is counting the heart beat of the Daphnia
is unaware of the concentration of caffeine in the solution it is in. This can prevent bias
due to the observers expectations. Studies have shown that observers expectations
can significantly influence the results.
Bibliography:
http://www.sciencebuddies.org/Files/3462/5/Pharm_img004.gif
http://www.ukessays.com/essays/biology/the-effect-of-caffeine-on-heart-rate-biologyessay.php#ixzz3Jsddsbmv
http://www.chem.purdue.edu/gchelp/molecules/caff.gif
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http://people.uwec.edu/janikds/biol312/lab%20discussion%20docs/312%20daphnia%20lab.pdf
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