ZOO 120 Animal Physiology

EXERCISE 1. Translocation of Materials

Name: Anna Mikaela M. Noble Date: Aug. 23

Section/Group # ST2L Group A Score ______________

DIFFUSION

Agar concentration 2% 0.048 cm/hr

5% 0.042 cm/hr
Temperature Room Temp (27C) 0.0535 cm/hr
Ref Temp (6C) 0.0363 cm/hr
Dyes Methylene blue 0.016 cm/hr
Congo red 0.042 cm/hr
Methyl orange 0.077 cm/hr

Describe the trend of the observed diffusion rates and discuss the effects of the following factors on the movement of
the 3 dyes:
a. Agar concentration
It was observed that there was a higher rate of diffusion in the 2% tubes whose mean diffusion rate was
calculated at 0.048 cm/hr, as it has lesser concentration of the agar in the mixture. On the other hand, the mean
diffusion rate of those in 5% agar was calculated at 0.042 cm/hr. The results imply that agar concentration is one
factor that can affect the process of diffusion. Therefore, the less concentrate the medium, the more space there
is for the molecules to diffuse into, resulting into a faster rate of diffusion (Monte, n.d.).

b. Temperature
The six test tubes were stored in the refrigerator with the temperature of 6 degree Celsius while the
remaining six test tubes was stored in a room with the temperature of 27 degree Celsius. As seen in the results obtained,
a higher mean diffusion rate, at 0.0535 cm/hr was observed for those stored in room temperature compared to the
mean diffusion rate of those stored in the refrigerator (0.0363 cm/hr). The higher the temperature, the higher kinetic
energy there is and the collision of molecules is increased. Furthermore, it induces conformational changes in protein
channels causing the increased passage of molecules as seen in the faster diffusion rate (Monte, n.d.).

c. Molecular Weight (discuss observed diffusion rates of the 3 dyes at 2% agar, room temperature condition only)
Three dyes were used in the experiment namely, Methylene blue, Congo red, and Methyl orange. Their
mean diffusion rates were calculated with Methyl orange (0.077 cm/hr) having the slowest diffusion rate, followed by
the Congo red (0.042 cm/hr), and finally, Methylene blue (0.016 cm/hr). In this test, the molecular weight of the dyes
used affected the rate of diffusion. With Congo red (696.69 g/mol) being the heaviest, followed by Methyl orange
(327.24 g/mol), and by Methylene blue (320.00 g/mol). Considering their molecular weights, the results show that the
dyes with the larger molecular size pass through the pores of the semipermeable membrane slower than those with a
smaller molecular size. Thus, the higher the molecular weight, the slower the diffusion rate (Randall, 1998).
Colloids are heterogenous substances that depend on the size of its particles. This dictates that colloids
resist flow as the particles of the substance adhere to the water molecules in the agar, resulting in a slower diffusion
rate. Therefore, the smaller the molecular size, the more stable the particles are. This is observed in the experiment as
the fastest dye to diffuse was methylene blue (0.016 cm/hr; 320.00 g/mol) (Monte, n.d.).

MOVEMENT ACROSS MEMBRANE

How did the addition of formalin and boiling treatments affect the diffusion of Congo Red molecules in the yeast cells?

Relative number of stained yeast cells exposed to different treatments

Test tube Treatment Cells stained*
A Formalin 50%
B Heat All cells stained
C Control 25%

Living yeast cells use active transport- which is the movement of substances across a membrane from a
region of low concentration to a region of higher concentration with the use of ATP- to keep the dye particles from
entering its membrane (Campbell, 2008). In the experiment, the control showed little staining as the plasma membrane
of the untreated yeast cells were less permeable to the dye. By generating energy and gradients, the protein pumps in
the cell membrane were able to pump out the dye particles. For test tubes A and B, both the addition of formalin and
heating has a direct effect on the permeability of the cell. Formalin is an organic solvent that dissolves the cell
membrane, thus destroying its selective permeability. On the other hand, the yeast suspension that was subjected to
boiling showed that all cells were stained as it denatured the proteins of the cells (Hill, 2012). Once the protoplasm
and membrane were destroyed, diffusion- which requires no energy expenditure- could take place. As dead cells
cannot generate energy and maintain gradients, the mechanism of the protein pumps that keep the dye out were all
denatured. Thus, the molecules of the dye moved from an area of higher concentration outside the cells and into an
area of lower concentration inside the cells (Randall, 1998).

OSMOSIS IN BIOLOGICAL MEMBRANES

Weight (g) of eggs placed in different solutions for 24 hours

Solutions Day 0 Day 1 Day 2
 Water 86.04 g 26. 49 g Membrane burst
Vinegar 77.14 g 84.00 g 84. 87 g
Syrup 83.02 g 85.16 g 84. 57 g

How did osmosis happen in the different solutions after Day 1? What happened after replacement of solutions with
distilled water on Day 2? Discuss.
Osmosis is the passive transport of water across a selectively permeable membrane. In this process, water
can travel in different ways. In a hypotonic solution, the extracellular fluid has a lower concentration than the
intracellular fluid causing the water to diffuse into the cell until equilibrium is achieved. In a hypertonic solution, the
concentration of the extracellular fluid is higher than the intracellular fluid, it causes water to diffuse out of the cell
until the equilibrium is reached. Finally, in an isotonic solution, water diffuses in and out of the cell such that there is
no net movement (Hill et al., 2012).

As the data shows, the weight of the egg decreased from 85.16 g to 84.57 g in the syrup solution, which
leads us to the conclusion that the water molecules were released in the syrup. Thus, the syrup served as the
hypertonic solution in this set up. Syrup has a high concentration of dissolved sugar molecules, giving it a high density.
Albeit dissolved, these molecules are still too large for it to pass through the cell’s membrane via passive transport.
However, water molecules can easily move through the membrane. Therefore, the water moved from inside the egg
to the surrounding solution until equilibrium is reached, causing the egg to decrease in weight (Kramer, 2012).
Vinegar is a liquid that is primarily made up of acetic acid and water, which served as the hypotonic solution.
The weight of the egg increased from 84.00 g to 84.87 g. Osmosis caused some of the vinegar to permeate into the
egg, causing it to enlarge.
The egg submerged in water exhibited a high degree of permeability as water molecules can pass through
selective channels. The concentration of water inside the egg has lower concentration than that of the surrounding
water. To achieve equilibrium, the water molecules from the outside permeated into it, resulting to an enlarged egg as
observed on day 1. This tells us that water is a hypotonic solute in the first day that imbibed water into the egg.
However, a plasma membrane can only expand to a certain limit. upon observation on the second day, the egg
appeared to have bloated and exploded as the threshold of the egg for water was reached.

OSMOTIC PRESSURE IN RBC

Hemolyzing concentrations and isotonic coefficient i of different solutions

Solutions [Hemolyzing] i
KCl 0.10M 0.5M
CaCl2 0.02M 0.1M
Na2SO4 <0.02M 0.05M
Compare observed i values of KCl, CaCl2, and Na2SO4. How does each solution affect the osmotic pressure leading to
hemolysis of RBCs?

Electrolytes such as KCl, CaCl2, Na2SO4 form ions in aqueous solutions. The results show us that KCl has the
greatest isotonic coefficient (0.5), followed by CaCl2 (0.1), and Na2SO4 (0.05). when contrasted to their hemolyzing
concentration, it can be seen that there is an inverse relationship. This implies that electrolytes exhibit hemolysis at
lower concentrations as they breakdown into ions that exert greater osmotic pressure. Additionally, it can be inferred
that red blood cells have lower hemolyzing concentrations in electrolytes than non- electrolytes (Kramer, 2012).

What is the role of sucrose in the experiment?

Sucrose was used in the experiment as it does not form ions in solutions. When added to solution, it
produces 1 mol of sucrose particle unlike electrolytes such as KCL that produces 2 mol of particles (Kramer, 2012).

Why are osmoregulation and osmotic balance important in animals?

Osmotic regulation in animals refer to the maintenance of a nearly constant concentration of electrolytes
and water balance across membranes within the body fluids such as the blood plasma and cytosol. Animals exchange
water and nutrients by eating and drinking. To regulate osmotic pressure, excretion in the form of sweat, urine, and
feces are performed by biological organisms in order to avoid the accumulation of toxic substances. As blood plasma
is a fluid component, the maintenance of a constant blood pressure is important in maintaining a healthy blood
pressure. It also maintains homeostasis, a state of equilibrium with the cell’s surrounding environment which allows
the cell to perform its functional properties (Hill et al., 2012).

References:
Hill, Richard W et al. (2012). Animal physiology -- 3rd ed. p. cm.
Campbell, NA and JB Reece. 2007. Biology. 8th ed. San Francisco: Benjamin Cummings.
Kramer, E. M., & Myers, D. R. (2012). Five popular misconceptions about osmosis. American Journal of Physics, 80(8),
694–699. doi: 10.1119/1.4722325

Monte, H. D. Translocation of Materials. Retrieved from

https://www.academia.edu/20812759/Translocation_of_Materials
Randall, D., Burggren. W., French, k., & Eckert, R. (1998). Eckert animal physiology: Mechanisms and adaptations. New
York: Freeman