Acknowledgments

We thank the University of Notre Dame

Biology Department, Dr. Mark Olsen,
Graduate TAMatt, and Undergraduate TAs
Michael and Linnea.
Introduction
P. Clarkii has a neurogenic heart that is controlled by nervous input, and can therefore be affected by exposure to
different chemicals. Neurogenic hearts are different from myogenic hearts that are controlled by muscle., and are
therefore less susceptible to changes in its chemical environment.
We chose to study the effects of ethanol and caffeine on the heart rate of P. clarkii because in myogenic hearts
like the human heart they have opposite effects, with ethanol acting as a depressant and caffeine acting as a
stimulant. However, the effects of some chemicals on heart rate in neurogenic hearts are reversed from their
effects on myogenic hearts. In a study by Judd et al. it was shown that caffeine has a depressing effect on the
neurogenic heart in P. clarkii, lowering its heart rate, which is the opposite of its effect on myogenic hearts such
as the human heart. There were no previous studies on the effects of ethanol on heart rate in P. clarkii.
The objective of our study was to determine if ethanol and caffeine had depressing or stimulating effects on
the heart rate of P. clarkii, and if the dosage of the chemical exposure had a correlation with the magnitude in
change of heart rate. We predicted that ethanol would act as a depressant, decreasing heart rate, and caffeine
would act as a stimulant, increasing heart rate, as in myogenic hearts. We conducted our study by first exposing
P. clarkii to normal saline solution and recording its heart rate, and then exposing the same P. clarkii to one of
our chosen chemicals in varying concentrations and recording the heart rate again. We then calculated the
percent change in heart rate after chemical exposure and ran an ANOVAtest to check for significance.
Materials and Methods

Vivisection was performed on crayfish samples obtained from the University of Notre Dame Biology
department. These had been anesthetized on ice until largely motionless. The thoracic ganglion was severed and
the legs, abdomen and head were removed. We placed each sample in saline solution and took its heart rate over
a minute. We recorded this value in beats per minute. We then took a second heart rate in the same manner but
adding out test solutions dropwise onto the sample beforehand. The concentrations of chemical we used were 5,
10, and 20 mM caffeine and 20, 40 and 80 mM ethanol. Each concentration was tested on three different
samples. We also included a control group that was not exposed to any chemical.
Results
7 out of 9 of the crayfish exposed to caffeine experienced a decrease in heart rate (Fig. 3).
All crayfish exposed to ethanol experienced a decrease in heart rate (Fig. 4).
None of our results can be viewed as statistically significant.
For caffeine: P=0.589
For ethanol: P=0.0837
This can be interpreted as a non-significant trend.
Conclusions

Ethanol
Ethanol showed a decrease in heart rate in all trials. An ANOVAp value of .0837 was calculated which does not allow
us to confirm our hypothesis that ethanol decreases heart rate in P. clarkii (Fig. 3). This p value shows a non-
significant trend with regards to the change in heart rate. With a larger sample size a significant result will likely be
possible to show that ethanol decreases heart rate in P. clarkii. If this change produces a significant result, we could
examine if ethanol works the same in P. clarkii as in humans because it causes the same decrease in heart rate in both
(Weng et al., 1999).
Caffeine
Caffeine caused a decrease in heart rate in 7 out of 9 trials which would seem like the opposite of our predicted
hypothesis that caffeine would increase heart rate in P. clarkii. An ANOVAp value of 0.589 was calculated which
means that the decrease in heart rate which we observed was not statistically significant (Fig. 4). Amuch larger
sample size would likely be needed to increase the significance and get rid of the effects of several large outliers in
our data. Astatistical test could also be run to identify these outliers. The seemingly unexpected decrease in heart rate
observed in the P. clarkii after application of caffeine is actually supported by a study by Judd et al (2007). Because of
this study and our results we would change our hypothesis that caffeine would increase heart rate for future
experiments.
Improvements
In order to improve the results of our experiment We should monitor our controlled variables more closely, such as
the amount of time on ice and the temperature of the saline solution. We should also increase our sample size for all
concentrations of ethanol and caffeine as well as the sample size of our control in order to produce more significant
results.
Future Studies
In order to understand the neurogenic hearts of P. clarkii better we could examine the effects of a chemical which
increases heart rate in P. clarkii. We could examine the effects which combinations of chemicals have on the heart
rate. For example, we could see if the effects of 2 chemicals which decrease heart rate compound their effects or we
could see if the effects of 2 chemicals with opposing effects cancel each others effects.
The Effects of Ethanol and Caffeine on the Heart Rate of Procambarus Clarkii
Patrick Bottone, Brian Roddy, John Salomone
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Fig. 2. P. clarkii body diagram






Fig 3. Comparison of different concentrations of ethanol on the heart rate of P. clarkii . Heart rate was taken over one minute. These represent the average percent change.
Each trial showed a decrease in heart rate.
Fig. 1. P. clarkii specimen

Fig. 4. Comparison of different concentrations of caffeine on the heart rate of P. clarkii. Heart rate was taken over one minute. These
represent the average percent change. All but one trial concentration decreased the heart rate on average.