Results

Effect of Toxin A on Paramecium Velocity

100.00
90.00
80.00
70.00
60.00
50.00 Toxin A
Average Velocity (ms-1) 40.00 Control
30.00
20.00
10.00
0.00

Time of measurement (seconds)

Figure 1: Effect of Toxin A on Paramecium Velocity: The average velocity the

average of the 8 Cell Motility lab sessions measurements - of Paramecia over a
period of 1800 seconds, in a solution containing toxin A (Ni 2+ ions), methyl cellulose
and the control measurement - without toxin. Paramecia velocity drops from the baseline
velocity to 0 ms-1, over the period of 1800 seconds.
Effect of Toxin A on the number of stationary Paramecia
35.00

30.00

25.00

20.00
Control
Number of stationary cells 15.00
Toxin A
10.00

5.00

0.00

Time of measurement (seconds)

Figure 2: Effect of Toxin A on the number of stationary Paramecia: The

average number the average of the 8 Cell Motility lab sessions measurements - of
stationary Paramecia over a period of 1800 seconds, in a solution containing toxin A
(Ni2+ ions), methyl cellulose and the control measurement without toxin. The number
of stationary cells which was 0 in the control setting, increases to 29 in 1800 seconds.

Effect of Toxin B on Paramecium Velocity

100.00
90.00
80.00
70.00
60.00
50.00 Control
Average Velocity (ms-1) 40.00 Toxin B
30.00
20.00
10.00
0.00

Time of measurement (seconds)

Figure 3: Effect of Toxin B on Paramecium Velocity: The average velocity

the average of the 8 Cell Motility lab sessions measurements - of Paramecia over a
period of 1800 seconds, in a solution containing toxin B (alcohol), methyl cellulose,
and the control measurement - without the toxin. Paramecium velocity drops from the
baseline velocity to 0 ms-1, over the period of 1440 seconds.

Effect of Toxin B on the number of stationary Paramecia

30.00
25.00
20.00
Control
15.00 Toxin B
Number of stationary cells 10.00
5.00
0.00

Time of measurement (seconds)

Figure 4: Effect of Toxin B on the number of stationary Paramecia: The

average number the average of the 8 Cell Motility lab sessions measurements - of
stationary Paramecia over a period of 1800 seconds, in a solution containing toxin B
(alcohol), methyl cellulose and the control measurement without toxin. The number
of stationary cells increases to 25 over the period of 1800 seconds.

Effect of Toxin C on Paramecium Velocity

100.00
90.00
80.00
70.00
60.00
50.00 Control
Average Velocity (ms-1) 40.00 Toxin C
30.00
20.00
10.00
0.00

Time of measurement (seconds)

Figure 5: Effect of Toxin C on Paramecium Velocity: The average velocity the

average of the 8 Cell Motility lab sessions measurements - of Paramecia over a
period of 1800 seconds, in a solution containing toxin C (unknown compound),
methyl cellulose, and the control measurement - without the toxin. Paramecia velocity
drops to 30ms-1 in 900 seconds, and then increases to the value of 90 ms-1 -the same velocity
as in the control setting- in another 900 seconds.

Effect of Toxin C on the number of stationary Paramecia

18.00
16.00
14.00
12.00
10.00
Control
8.00
Number of stationary cells Toxin C
6.00
4.00
2.00
0.00

Time of measurement (seconds)

Figure 6: Effect of Toxin C on the number of stationary Paramecia: The

average number the average of the 8 Cell Motility lab sessions measurements - of
stationary Paramecia over a period of 1800 seconds, in a solution containing toxin C
(unknown compound), methyl cellulose and the control measurement without
toxin. The number of stationary cells increases to 15 over 900 seconds and then decreases to 0 in
another 900 seconds.
Conclusions

The effects of toxin A on paramecium velocity are due to the effects of Ni2+ on the
axonema of the cilium. Ni2+ affects the dynein motors of the cilium, inhibiting their ATPase
activity. The toxin inhibits the translocation of microtubules facilitated by the dynein motors,
which in turn stops the bending of the cilia, and therefore their beating. All cilia are stopped in a
hands down position curved towards the posterior end of the cell. Of the 22S and 14S species
of dynein, Ni2+ has a much greater effect on the 14S species. The 22S species can function at up
to 10 times the concentration needed to inhibit the activity of the 14S species. The concentration
needed to stop the movement of cilia must be above 0.1 mM, and the concentration of Ni2+ toxin
in our experiment was 0.1 mM too (Larsen and Satir, 1991).
The effects of toxin B on paramecium velocity are due to the effects of alcohol on cilia.
Alcohol has ciliostasis effects, known of after studying its effects on respiratory ciliated cells. In
small concentrations (220 mM) alcohol has stimulating effects on cilia motility, but in higher
concentrations (> 400mM), it has an inhibitory effect. Also, prolonged exposure to alcohol
causes a desensitization of the cilia, no longer being able to respond to external stimuli (Sisson,
2007). Although the concentration in our experiment was 19mM, which should be too low, the
toxin still had a strong effect. This could be attributed to the alcohol not being in blood,
metabolised by the liver, but in direct contact with the paramecia. Alcohol acts on the outer
dynein arm, targeting specific components, like the and heavy chain motor subunits, reducing
beating frequency. (Yang et al., 2015).
Comparing the data from toxins A and B, alcohol has a quicker effect than Ni2+ on
reducing the velocity of paramecia, reaching its full effect in 1440 seconds vs. 1800 for the Ni2+.
However, toxin A affected on average 4 more paramecia than toxin B did.
Unlike the effects of toxins A and B, which were both irreversible, the effects of toxin C
reached their peak at 900 seconds. After that, the effects of toxin C started to revert, and the
paramecia regained their control-setting velocity after another 900 seconds. Also, the amplitude
of toxins C effect was lesser then the amplitude of the other toxins effects. The velocity only
dropped to 30 ms-1 vs. 0 ms-1 in the case of the other 2 toxins, and the number of stationary
cells only reached 15 vs. 25-29 for toxins A and B. This can suggest that toxin C is a compound
that can be metabolised by paramecia, and might not directly target the cilia, but the whole cell,
allowing the process of metabolising.
These results pose environmental concerns, as heavy metals like Ni and alcohol, if they
get into waters, will stop the eukaryotic cilia motility, affecting both microorganisms and more
evolved species alike, leading to an ecological imbalance.
 References

Larsen, J. and Satir, P., 1991. Analysis of Ni(2+)-induced arrest of Paramecium

axonemes. Journal of Cell Science, 99(1), p.33 Available at:
<http://jcs.biologists.org/content/99/1/33.abstract> .

Sisson, J.H., 2007. Alcohol and Airways Function in Health and Disease. Alcohol (Fayetteville,
N.Y.), 41(5), pp.293307 Available at:
<http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2081157/> .

Yang, F., Pavlik, J., Fox, L., Scarbrough, C., Sale, W.S., Sisson, J.H. and Wirschell, M., 2015.
Alcohol-induced ciliary dysfunction targets the outer dynein arm. American Journal of
Physiology - Lung Cellular and Molecular Physiology, 308(6), pp.L569L576. Available at:
<http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4360061/> .