Joseph Lee, Nicholos Joseph, Rohan Kondetimmanhalli

Per. 6 and 7
April 5th, 2013
Brine Shrimp Lab #1
Purpose: To determine how salinity affects the development of brine shrimp
Background: Adaptations to the land and other benefits allow organisms to survive in the

ecosystem/environment in which they are placed. The environment selects certain individuals to

have a greater chance of reproducing. However, environmental conditions change, like salinity of

waters. A few mass extinctions have already occurred throughout Earths history, causing

speciation to take place. The new species eventually develop characteristics that aid in surviving

in new environments. One example is the brine shrimp. They dwell in salt water lakes and are

greatly affected by changes in salinity. Under ideal conditions, the brine shrimp will go through

development very quickly, while changes in the environment may force the shrimp to dwell in

their eggs. Changes in the salinity of the water will enable us to study the ideal conditions and

the effects of low and high salinity on the shrimp.

Hypothesis: Increased salinity will decrease the development time of brine shrimp.
Materials:

Brine shrimp eggs Double-sided tape Scale

Scissors Purified water 5 beakers
5 pipets 5 petri dishes Sodium chloride
1 stereomicroscope Permanent marker Lab spoons
Paintbrush Stirring rod Weigh boats
5 microscope slides Graduated cylinder
Procedures:
Day 1

1. Prepare and label five beakers of 30-ml salt solutions, using sodium chloride and

dechlorinated water. The solutions should be 0%, 0.5%, 1%, 1.5%, and 2% NaCI. Use the

space in the Analysis section that follows to determine what mass of sodium chloride to

add to each solution.

2. Label five petri dishes, 0%, 0.5%, 1%, 1.5%, 2%.
3. Use a graduated cylinder to measure 30 ml of 0% salt solution and pour it into the petri

dish labeled 0%.

4. Likewise measure and pour 30 ml of each of the remaining concentrations into their

corresponding petri dishes.

5. Measure and cut 1.5 cm of double-sided tape.
6. Stick the double-sided tape to a microscope slide
7. Lightly touch the paintbrush to the side of the bag containing the brine shrimp eggs. Your

goal is to collect only approximately 20 eggs on the brush. You do not want to cover the

tip of the brush in eggs.

8. Dab the paintbrush onto the tape on the microscope slide.
9. Examine the slide under a stereomicroscope. Count the number of eggs on the slide and

record this number in Table 1 under "0 hours."

10. Place the microscope slide in the 0% petri dish, tape-side up, and place the lid on the

dish.
11. Follow steps 5-11 for the remainin_g slides and dishes, until you have prepared five

microscope slides of eggs, recorded the numbers in Table'{ and placed each slide in its

appropriate salt solution.

12. Allow the dishes to sit at room temperature undisturbed for 24 hours.
Day 2
1. Examine one petri dish with a stereomicroscope
2. Count the number of swimming brine shrimp. With a pipet, gently catch them and move

them into another container-your instructor may have prepared a beaker of saltwater for

this purpose. Try not to remove excess water with the pipet. Record the number of

swimming shrimp in Table 1 under "24 Hours."

 3. Count the number of dead or partially hatched shrimp and record this number in Table 1

under "24 Hours."

4. Count the number of unhatched eggs and record this number in Table 1 under "24 Hours."
5. Repeat steps 1-4 for each of the petri dishes. Use a separate pipet for each petri dish.
Day 3
1. Examine each petri dish with a stereomicroscope
2. Count the number of swimming brine shrimp. With a pipet, gently catch them and move

them into a beaker. Record the number of swimming shrimp in Table 1 under "48 hours."
3. Count the number of dead or partially hatched and record this number in Table 1 under

"48 hours."
4. Count the number of unhatched eggs and record this number in Table 1 under "48 hours."
5. Repeat steps 1-4 for each of the petri dishes. Use a separate pipet and beaker for each

petri dish.
6. Clean up petri dishes.

0 hours 24 hours 48 hours

% # Dead or # Dead or
NaC Partially # Partially Hatching
Dish # l # Eggs # Eggs Hatched # Swimming Eggs Hatched # Swimming Viability
1 0 59 46 6 7 30 21 8 13.56%
2 0.5 43 35 3 5 26 9 8 18.60%
3 1 53 40 2 11 22 7 24 45.28%
4 1.5 42 32 2 8 11 6 25 59.52%
5 2 39 11 5 23 2 7 30 76.92%

Results: As evidenced by the clear trend, an increase in concentration of NaCl in the solution in

which the brine shrimp breed results in a higher level of hatching viability. With no NaCl, the

viability was just 13.56%, but this percentage skyrocketed to 76.92% in the 2% NaCl solution.

Conclusion: The proper habitat for brine shrimp to breed would be in salt water, such as in

oceans or salinized lakes. Freshwater is not a viable habitat, as it results in high levels of failed

birth for the brine shrimp. This is probably due to evolutionary patterns which resulted in

placement of brine shrimp in primarily salt water settings, and caused the brine shrimp to adapt
to that environment in time over many generations. In addition, salt water often harbors a variety

of nutrients, such as plankton, krill, and other microscopic organisms, which are not often found

in freshwater lakes.

Brine Shrimp Lab #2

Purpose: To analyze how temperature of the salt solution affects the development of brine shrimp
Materials: 5 beakers

150 mL of purified Hot plate Tape

Brine shrimp eggs Microscope
water 5 petri dishes Paintbrush
Thermometer 5 microscope slides
Procedures:
Day 1
1. Measure out 30 mL of purified water for each beaker and pour into the beakers.
2. Attach a piece of double sided tape to each slide.
3. Use a paintbrush to collect approximately 50 brine shrimp eggs and brush them onto the

tape.
4. Heat each beaker to 21,23,25,27, and 29 degrees Celsius respectively.
5. Empty the water into each respective petri dish and place slides into the dishes.
6. Allow dishes to sit for 24 hours.
Day 2
1. Count the number of dead, hatched, and eggs in the petri dish and record.
2. Remove the dead and the hatched shrimp from the dishes.
3. Heat the petri dishes back to their original temperatures.
4. Allow the dishes to sit for 24 hours.
Day 3
1. Repeat the steps listed for Day 2.
2. Clean up the lab.

0 hours 24 hours 48 hours

# Dead or # Dead or
Temp Partially # Partially Hatching
Dish # (C) # Eggs # Eggs Hatched # Swimming Eggs Hatched # Swimming Viability
1 21 56 43 2 11 30 5 21 37.5%
2 23 51 45 1 5 35 7 9 17.65%
3 25 43 38 2 3 27 10 6 13.95%
4 27 49 46 2 1 26 20 3 6.12%
5 29 57 55 1 1 32 18 7 12.28%

Results: Temperatures nearing 30 degrees Celsius cause brine shrimp to perish or be unable to

hatch from their eggs. For instance, 29 degrees had a hatching viability of just 12.28%,

indicating that the temperature is obviously not optimal. Even 21 degrees Celsius, at 37.5% for

its hatching viability, suggests that optimal temperatures are probably slightly lower than the

ones provided.
Conclusion: Temperatures that extend high are not optimal for brine shrimp hatching or growth.

This makes sense, as an ocean environment is often cold due to indirect sunlight, and brine

shrimp have grown accustomed to such a setting. In addition, higher temperatures often lower
metabolism and activity, which is a fact supported by the data collected. Lower temperatures aid

in regulation for an optimal setting for growth of brine shrimp.