Tuhina Kazi

BIO 10200 Section 3EG2

Lab Partner: Nicholas Molohides

November 1, 2017

The Effects of Common Household Drugs on Circulatory Functions

Abstract

This experiment studies the effects of common household drugs on circulatory functions,

focusing primarily on how caffeine and nicotine affects pulsation rates in Lumbriculus

variegatus. Lumbriculus variegatus is an ideal specimen for this experiment due to their thin

cuticle and epidermis, which allows substances in their environment to easily penetrate and cause

a reaction with their circulatory system. This experiment will follow six levels of treatments,

three levels of increasing concentration of caffeine and three levels of increasing concentration

of nicotine, to see how varying concentration will affect pulsation rate.

Introduction

The circulatory system is a network of organs that are in charge of pumping blood,

nutrients, and hormones to the different areas of the body. It is essential to the body in order to

maintain homeostasis, optimal internal living conditions. Many things can affect the physiology

of our bodies, such as caffeine and nicotine. Caffeine is a common household drug that has the

ability to counter the effects of inhibitory neurotransmitters while nicotine mimics the effects of

the neurotransmitter, acetylcholine (Lab Manual, 2017). To see the effects of caffeine and

nicotine on one facet of physiology, pulsation rate in Lumbriculus variegatus will be observed. It
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is predicted that if the blackworms are administered the caffeine and nicotine, then the pulsation

rate will increase.

Lumbriculus variegatus, more commonly known as the blackworm, is a species of worm

that reside in Northern America and Europe in freshwater. The blackworm has a pair of lateral

commissural vessels in the anterior segment (Lesiuk et al. 1999). Commissural vessels are the

lateral hearts of earthworms that pumps blood from the dorsal to the ventral vessel (Crisp et al.

2010).

Due to the transparent bodies of the blackworm, pulsation rate can be easily tracked

through the use of a microscope. Since caffeine counters the inhibitory neurotransmitters and

nicotine mimics acetylcholine, it is hypothesized that both the caffeine and nicotine will increase

pulsation rate of the blackworms being observed. The null hypothesis of this experiment is that

neither the caffeine nor the nicotine will have an effect on the pulsation rate of blackworms.

Methods

To test the hypothesis, the variables must be identified. The independent variables of the

experiment are the different concentrations of caffeine (1mM, 3mM, and 10mM) and nicotine

(0.05mM, 0.25mM, and 1.0mM). The dependent variable is the pulsation rate of the blackworms.

The standardized variables are the number of worms per treatment (3 worms per treatment) and

the type of worms (Lumbriculus variegatus), the materials used to conduct the experiment, like

the microscope and pipets. In the experiment there were a total of six levels of treatments (the

different concentrations of caffeine and nicotine) and each level contained a sample size of three.

There was only one replication for this experiment.

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To conduct the experiment, a pipet was used to retrieve the blackworm and place it onto a

parafilm slide to record it’s basal pulsation rate (rate before treatment) for twenty seconds with

the aid of a stopwatch, which will be converted to beats per minute, while under the microscope.

Readings shall be taken three times with a thirty-second interval between each reading and then

averaged. This is repeated two more times for worm B and worm C. Worm A is then placed in

its designated treatment for at least fifteen minutes before getting briefly rinsed with spring

water. Pulsation rates are recorded again by following the same steps.

Results

Average Change in Pulsation Rate

(Caffeine)
Change in Mean Pulsation Rate

20

15
(#beats/min

10

0
0 2 4 6 8 10 12
Caffeine Concentration (mM)

Figure 1: The graph shows the average change in pulsation rate in blackworms for increasing

concentrations of caffeine where each point represents the mean value.

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Pulsation Rate in Caffeine

18
Change in Rate (#beats/min)

16
14
12
10
8
6
4
2
0
1 mM 3 mM 10 mM
Treatment

Figure 2: The graph represents the mean of the blackworms pulsation rate in all three caffeine

treatments. The error bars represent the standard deviation.

The graph shows the average change in mean of pulsation rate in each treatment

respectively. The average change for 1mM, 3mM, and 10mM are 4.45 (beats/min), 3.07

(beat/min), and 15.43 (beats/min) respectively. The error bars represent the standard deviation

for all three treatments. 1mM had a standard deviation of 0.042, 3mM had a standard deviation

of 0.304, and 10mM had a standard deviation of -0.037.

T-Table for Caffeine:

|t-calculated| 8.44

t-critical for 95% confidence 2.776

D.F. 4

Confidence Level >99.9%

Table 1: The table shows the t-test results of the comparison of changes in mean pulsation rate

within 10mM of caffeine.

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The table shows the absolute value of the t-calculated value, the t-critical value for a 95%

level of confidence, the degree of freedom in this experiment, and the confidence level in the

data collected for the experiment. The t-calculated value is 8.44, the t-critical value is 2.776, the

degree of freedom is 4, and the level of confidence for this experiment is over 99.9%.

Average Change in Pulsation Rate

(Nicotine)
25
Change in Mean Pulsation Rate

20

15
(#beats/min)

10

0
0 0.2 0.4 0.6 0.8 1 1.2
-5
Nicotine Concentration (mM)

Figure 3: The graph shows the average change in pulsation rate in blackworms for increasing

concentrations of nicotine where each point represents the mean value.

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Pulsation Rate in Nicotine

30
Change in Rate (#beats/min)

25

20

15

10

0
0.05 mM 0.25mM 1 mM
-5
Nicotine Concentration (mM)

Figure 4: The graph represents the mean of the blackworms pulsation rate in all three nicotine

treatments. The error bars represent the standard deviation.

The graph shows the average change in mean of pulsation rate in each treatment

respectively. The average change for 0.05mM, 0.25mM, and 1mM are -1.67 (beats/min), 13.83

(beat/min), and 20.67 (beats/min) respectively. The error bars represent the standard deviation

for all three treatments. 0.05mM had a standard deviation of -0.627, 0.25mM had a standard

deviation of 4.537, and 10mM had a standard deviation of 7.043.

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T-Table for Nicotine:

|t-calculated| 12.85

t-critical for 95% confidence 2.776

D.F. 4

Confidence Level >99.9%

Table 4: The table shows the t-test results of the comparison of changes in mean pulsation rate

within 1mM of nicotine.

The table shows the absolute value of the t-calculated value, the t-critical value for a 95%

level of confidence, the degree of freedom in this experiment, and the confidence level in the

data collected for the experiment. The t-calculated value is 12.85, the t-critical value is 2.776, the

degree of freedom is 4, and the level of confidence for this experiment is over 99.9%.

Discussion

By increasing the concentration of caffeine and nicotine, it was assumed that there would

be a change in the pulsation rate of Lumbriculus variegatus. According to the results, the

hypothesis is supported. The absolute value of the t-calculated value is larger than the t-critical

value in both t-tests for caffeine and nicotine. This indicates that the results from all treatments

have significant difference. From this information we are able to determine that our null

hypothesis can be rejected and that our alternative hypothesis is supported (Course Supplement,

2017).

Sources of error in this experiment can be attributed to human error in reading pulsation

rate. Demonstrated in Figure 1, between 1mM caffeine and 3mM caffeine, there is a decrease in

change in mean pulsation rate before it begins to increase once more between 3mM caffeine and
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10mM caffeine. A similar occurrence can be seen in Figure 4 where there is a small drop in the

change in mean pulsation rate going from 0mM of nicotine to 0.05mM of nicotine. Values after

0.05mM of nicotine then increase drastically.

Due to the fragile nature of the Lumbriculus variegatus, certain parts of the experiment

had to be redone because fragmentation had caused the worm to reproduce. This can be

considered a limitation as the results could have been skewed because of their hermaphroditic

properties.
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Literature Cited:

 City College of New York, 2017. Laboratory Manual for Biological Foundations II Bio

10200. New York, NY.

 Crisp, K. M., Grupe, R. E., Lobsang, T. T., Yang, X. (2010). Biogenic amines modulate

pulse rate in the dorsal blood vessel of Lumbriculus variegatus. Comparative

Biochemistry and Physiology Part C: Toxicology & Pharmacology, 151(4), 467-472.

doi:10.1016/j.cbpc.2010.02.003

 Lesiuk, N. M., & Drewes, C. D. (1999). Blackworms, Blood Vessel Pulsations & Drug

Effects. The American Biology Teacher, 61(1), 48-53. doi:10.2307/4450609

 City College of New York, 2017. Course Supplement for Biological Foundations I Bio

10100. New York, NY.