Temperature, pH and Salinity: Limiting Factors Affecting Range of

Tolerance
Objectives
1. To investigate the effects of temperature changes on the metabolic or respiratory
rates of fish.
2. Effects of changing pH and salinity levels on the growth, development and survival of
selected plants and animals.

Methodology
Temperature effects
1. Secure 3 wide-mouth bottles of uniform sizes and label as 1, 2 and 3.
2. Half-fill each bottle with tap water.
3. Using a laboratory thermometer, adjust the water temperature in bottle 2 with ice
water at around 10C - 15C and maintain the temperature during the entire
experiment.
4. Adjust the water temperature in bottle 3 with boiled water at around 35C - 40C
and maintain the temperature during the entire experiment.
5. Determine the water temperature in bottle 1 and this will serve as the control.
6. Place 1 catfish in each bottle (of uniform size) and leave for a minute.
7. Observe the rate of opercular movement for 3 minutes and record your
observations in tabular form. Make 3 replicates.
pH tolerances
1. Prepare 1L each of test solution (0.01M NaOH) adjusted to pH 2, 6, 7, 8 and 10.
2. Transfer (up to half mark) each solution in separate wide-mouth container and label
properly each container.
3. Carefully place 10 guppies of approximately the same size, age and health status in
each of the test solutions.
4. Observe the guppies within 15 minutes and take note of morphological and
behavioral responses such as swimming movements, breathing, change in body
color, etc.
5. At the end of 15 minute exposure period, count the number of dead guppies in each
test solution.
6. Determine the % mortality of guppies for each pH level.
7. Plot the mortality data with pH levels and analyze and interpret the results.

Activity 2: Limiting Factors affecting Range of Tolerance

Fundamental of Ecology (laboratory)
Department of Biology, College of Science
Polytechnic University of the Philippines
A.S. Coronado

Page 1

Salinity effects
A. Freshwater snails
1. Prepare 500 mL each of salt solutions at 2.5 ppt, 5.0 ppt, 10.0 ppt, 15.0 ppt
and 20.0 ppt.
2. Prepare 6 250 mL capacity plastic cups and fill with each salt solution until its
ream. Make a control using tap water. Reserve the remaining salt solution
for the next experiment.
3. Put 3 snails of uniform sizes in each container.
4. Cover each cup with nylon net and secure with rubber band.
5. Observe for 4 days and take note the behavioral responses of the snails in
each set-up.
6. Count the number of deaths for the entire duration of the experiment and
present your results in graphical form.
B. Corn
1. Soak for overnight about 100 corn seeds, adding a small amount of fungicide.
2. Secure 6 sets of Styrofoam lunch boxes and laden with 4 tiers of tissue paper.
3. Label each set corresponding to the 5 salt solutions and tap water as control.
4. Sprinkle each salt solutions and tap water correspondingly to the Styrofoam
boxes.
5. Transfer 10 ungerminated corn grains to each box and continue to sprinkle
the salt solutions daily for 5 days.
6. Observe the % germination of corn seeds after 5 days for each set-up.
7. Count the number of seeds successfully germinated (a portion of the embryo
is already visible from the coat).
8. Calculate % germination for each set-up using the formula:
(no. of seeds germinated / total number of placed seeds) x 100
9. Record all the values in tabular form and plot the values obtained with
salinity levels on the x-axis and % germination on y-axis.
10. After 5 days, measure also the root length (R) and the shoot length (S) for all
the germinated seedling in each set up.
11. Derive the root to shoot ratio (R/S) from the previous data and compare the
results. Record all the values to the nearest tenth of a millimeter. Plot the
obtained values.

Activity 2: Limiting Factors affecting Range of Tolerance

Fundamental of Ecology (laboratory)
Department of Biology, College of Science
Polytechnic University of the Philippines
A.S. Coronado

Page 2

Guide questions
1. Outline and briefly discuss the cellular and physiological effects of temperature change
on living organisms.
2. What is meant by Vant Hoffs rule? Is this true for both terrestrial and aquatic
ecosystems? Explain.
3. Differentiate the following terms: ectotherms, endotherms, poikilotherms,
homoitherms, oscomoconformers, osmoregulators, eurytopic and stenotopic.
4. Provide an explanation for the observed behavioral responses of fish to pH changes. In
general, what are the physiological effects of pH changes on living organisms? Would
you say that your observations typify the conditions on acidic lakes?
5. Discuss the relationship between salinity and mortality among freshwater snails.
Outline the cellular mechanism that could possibly explain the observed effects.
6. Provide a feasible explanation for the observed effect of salinity on % germination and
growth rate in corn seeds.

Activity 2: Limiting Factors affecting Range of Tolerance

Fundamental of Ecology (laboratory)
Department of Biology, College of Science
Polytechnic University of the Philippines
A.S. Coronado

Page 3