RESULTS AND DISCUSSION

The circulatory system (cardiovascular system), is an organ system that permits blood to
circulate and transport nutrients, ions, gases, hormones, and blood cells to and from the cells in
the body, providing nourishment and help in fighting diseases, stabilize temperature and pH,
basically maintaining homeostasis (Reese and Campbell, 2011). Systemic circulation is the part
of the circulatory system which carries oxygenated blood away from the heart to the body, and
returns deoxygenated blood back to the heart.
Two types of cardiovascular system occurs in organisms. One is a closed circulatory
system, which is exhibited by all vertebrates and a few invertebrates. Blood flows within vessels
or connecting tubes within different size and wall thickness, thus called by its name. It does not
normally fill the body cavities which occur in open circulatory systems (Sakurai, 2011).
As advancement of organisms evolve, so does the circulatory system. In this exercise
toads are used in order to picture out systemic circulation since it is very similar that of the
circulatory system in humans. It differs by having a three chambered heart allows for the
advancement of a double circuit. First is the pulmocutaneous circuit which involves acquiring
oxygenated blood from the lungs and skin which then enters the left atrium. The other is the
systemic circuit where blood is pumped to the body organs and returns to the heart. In this
experiment, effects of different chemicals and temperature on the flow of blood in the systemic
circuit is observed. It is made possible by the diffusion of liquids with different treatments into
the toads skin. Movement of blood across vessels were made possible to be seen using a
microscope.

Figure 1. Amphibian blood circulation (Lifted from: Reese and Campbell, 2011).
Intervention of test solutions were carried out by diffusing across web epithelium. Its
effects largely depend upon the conditions prevailing at the time of administration, the dose used
and the duration itself. Four chemicals were applied in the toad; caffeine, acetic acid, nicotine,
and alcohol. These can act as vasoconstrictors and vasodilators. The latter, relax the smooth

muscle in blood vessels which causes dilation. The former, contract the smooth muscle in blood
vessels which causes constriction.
Caffeine has been recognized as the most widely used psychoactive drug in the world
(Gilbert, 1976). The said chemical has a wide range of pharmacological effects and is positively
associated with stimulation of cardiovascular system and central nervous system (CNS),
glandular tissue, action on the kidney to produce diuresis, and relaxation of smooth muscles
(Amer Suleman and Siddiqui, n.d.). At high concentrations (0.5 to 1 mM) it interferes with the
uptake and storage of Ca2+ by the sarcoplasmic reticulum in striated muscle, specifically resulting
to spastic contractions of the heart hastening the blood flow. Caffeine is a CNS stimulant of the
methylxanthine class of psychoactive drugs (Nehlig, 1992). Methylxanthines play a role in
translocation of intracellular calcium, increasing accumulation of cyclic nucleotides, and
obstruction of receptors for adenosine. Caffeine affects the blood flow by hastening it through
dilation of peripheral vessels (Amer Suleman and Siddiqui, n.d.).
Acetic acid is utilized in anaerobic respiration. It can be observed when a man undergoes
a physical activity then after sometime a buildup of lactic acid on the muscle reaches a high level
during the process. When there is shortage in oxygen available for the muscle to utilize during
the breakdown of glucose for energy, acetate is made where its build up is faster than burning it
off in your bloodstream thus creating a lower pH level and causes dilation in vessels. This
change in pH in peripheral capillaries cause dilation to allow more oxygen in the blood to
compensate the oxygen deficiency in the vessels due to acidosis, increasing heart rate, thus a
vasoconstrictor in cardiac capillaries.
Nicotine was also tested in the toads webbed foot. Nicotine initially mimics
parasympathetic actions where it inhibits the heart from contracting, but eventually blocks
parasympathetic activities and has the opposite effect which accelerates the heartbeat. According
to the research conducted by Horimonto and Koyama (1982), the mean pulsatile amplitude
significantly increased after immersing the toads web in the nicotine solution. After the nicotine
enters the body, it causes different changes like the release of hormones like adrenaline which
acts as a vasoconstrictor (Black et. al., 2001). It has an effect on the systemic circulation making
it harder for the heart to pump through the constricted arteries - and it causes the body to release
its stores of fat and cholesterol into the blood.
In toads it has been recorded that 50% alcohol diffusing into the webbed foot determines
an immediate though moderate constriction (Dixon, 1907). This phenomenon induces blood flow
to slow down. According to Dixon (1907) it is clear that alcohol possesses two entirely different
actions, it excites in small doses but depresses in larger ones. In the laboratory 95% of alcohol
was used. Alcohol administered in normal animals in large doses slows the heart through the
vagus nerve. It is a vasodilator which is an agent that induces vasodilation or the widening of
lumen of the blood vessels. In moderate doses alcohol causes dilatation of the superficial vessels
and some slight constriction, which after large doses is followed by dilatation, of internal vessels
(Dixon, 1907). It inhibits the antidiuretic hormone release or the impairment of vasoconstriction.
The volume of blood brought to the skins surface increases, leaving a warm feeling.

Temperature also have its own effects in systemic circulation. These were studied in
humans using laser Doppler flowmetry (Song et al., 1989). When skin was heated (40 degrees C)
red cell flow in the arms increased, when subjected to much lower temperature (5 degrees C)
flow momentarily decreased, but soon increased, surpassing the pre-cooling level. Then
continued to decline when cooling was prolonged more than 15 minutes.
On one hand warm water increases heart rate and causes vasodilation in peripheral
capillaries, allowing lots of blood to flow near the surface and heat is lost. On the other hand,
cold water induces constriction in which blood is kept away from the surface, narrowing the
blood vessels leading to the skin capillaries. In effect, very little blood flows through the
capillaries minimizing the loss of heat from the skin.
Amphibian Ringers solution played a vital role in carrying out the experiment in one
toad. It served as the washing agent to bring back homeostasis in the toads webbed hind foot.
The solution maintains an isotonic solution, providing enough moisture for the toad. Without it,
data to be gathered in the experiment will be extremely flawed. Note that the effects of the test
solutions were compared with the initial blood flow in vessels of the toad rinse with Ringers
solution.
Table 1. Blood flow of three different toads after subjected to different test solutions.
Test Solutions
Caffeine
Acetic Acid
Nicotine
Alcohol (95%)
Warm Water
Cold Water

1
Faster
Faster
Slower
Slower
Faster
Slower

2
Faster
Faster
Slower
Faster
Faster
Slower

3
Faster
Faster
Faster
Faster
Faster
Slower

The trend between the three toads experimented in applying water at different
temperatures are exactly the same as the ideal results. At higher temperature blood flow went
faster while at low temperature it slowed for a given time and then immediately rinsed with
amphibians Ringers solution.
Toads were under a lot of stress by testing different chemicals in its webbed hind foot. So
inconsistencies in the results were a staple. As for nicotine, the third group could have been not
observant or the toads web was not washed thoroughly with Ringers solution leaving a faster
effect in the blood flow in the peripheral vessels. Alcohol also met this problem for the first
groups toad. Alcohol has also a mechanism to initially constrict vessels then dilates it at high
doses. The result might have been to immediate for the group to conclude that it slowed down
the blood flow in the vessel.
For caffeine and acetic acid, effects were clearly seen in all toads experimented. All were
aligned with the expected results. Blood flow were faster than that of the normal.
REFERENCES

Amer Suleman, M. D., and Siddiqui, N. H. Effects of Temperature and Drugs on Toad Heart.
Black, C.E., N. Huang N. P.C. Neligan, R.H. Levine, JE Lipa, S. Lintlop, C.R. Forrest, and C.Y.
Pang. 2001. Effect of nicotine on vasoconstrictor and vasodilator responses in human skin
vasculature. Am J Physiol Regul Integr Comp Physiol 281 (4):1097-104
Dixon, W. E. 1907. The action of alcohol on the circulation. The Journal of physiology, 35(4),
346-366.
Gilbert R. M. 1976.Caffeine as a drug of abuse. In: Gibbins R. J., Israel Y., Kalant H., Popham R.
E., Schmidt W., Smart R. G. eds. Research advances in alcohol and drug problems. vol. 3. New
York: John Wiley & Sons.
Nehlig A, JL Daval, and G Debry. 1992. Caffeine and the central nervous system: mechanisms of
action, biochemical, metabolic and psychostimulant effects. Brain Res. Brain Res. Rev. 17 (2):
13970.
Horimoto, M. and T. Koyama. 1982. Acute microcirculatory response to nicotine in frog web.
The Japanese journal of physiology, 32(5), 771-782.
Reece, J. B., and N. A. Campbell. 2011. Campbell biology. Boston: Benjamin Cummings /
Pearson.
Song, C. W., L. M. Chelstrom, S. H. Levitt, and D.J. Haumschild. 1989. Effects of temperature
on blood circulation measured with the laser Doppler method. International Journal of Radiation
Oncology* Biology* Physics, 17(5), 1041-1047.