Core Practical 12: Investigate the effects of temperature on the

development of organisms (such as seedling growth rate or brine

shrimp hatch rates), taking into account the ethical use of
organisms

Objectives
• To investigate the effect of temperature on the hatching success of brine shrimp /
germination success of seeds
• To know and understand how to use a wide range of apparatus, materials and
techniques safely and appropriately
• To understand ethical issues related to the use of living organisms in investigations
Safety
• Take care when handling glassware.
• Wear eye protection.
• Do not handle lamps, plugs, sockets or switches with wet hands.
• Filament lamps will get hot enough to burn skin and may explode if splashed with
water.
• Wash hands with soap and water after the practical work.
Maths skills
• Find arithmetic means.
• Understand the terms mean, median and mode.
• Plot two variables from experimental or other data.
Equipment
Brine shrimp
• brine shrimp egg cysts
• 2 g of sea salt
• dechlorinated water
• 100 cm3 measuring cylinder
• six beakers (250 cm3)
• stirring rod
• waterproof marker
• water baths (25 °C, 30 °C and 35 °C)
• access to refrigerators set at 15 °C and 20 °C
• thermometer
• forceps
• lamp or bright light source
• pipettes
• graph paper
Seeds
• seeds (mustard or cress seeds work well)
• distilled water
© Pearson Education Ltd 2019. Copying permitted for purchasing institution only. This material is not copyright free.
Practical activities have been safety checked but not trialled. Users may need to adapt the risk assessment
information to local circumstances. This document may have been altered from the original. 1
 • 100 cm3 measuring cylinder
• Petri dishes
• cotton wool
• waterproof marker
• incubators set at 25 °C, 30 °C and 35 °C
• access to refrigerators set at 15 °C and 20 °C
• forceps
• thermometers
• graph paper
Ethics
There are certain ethical considerations you must take into account when experimenting
on animals or carrying out dissections. The benefits (in terms of advancing knowledge)
must be balanced against any potential harm to a living thing. One of these investigations
involves examining the effects of variables on the hatching success of an organism (figure
A), and handling the live organisms experimentally. These skills and knowledge cannot
fully be provided by other means.
As a student, you have a responsibility to maintain an ethical approach to the use of
animals. This includes a responsibility to derive the maximum possible learning benefit
from any use of animals or animal tissues in your classes. You also have a duty of care for
the welfare of any live animals you use.

figure A Brine shrimp

Procedure
When choosing which living organism to work with, availability is a key consideration. For
this investigation, you must consider both the ethics and the practicalities of obtaining the
organism.
It is recommended that you carry out this investigation using brine shrimp because this will
require a higher degree of practical skill. However, you can work with seedlings instead, if
necessary. Whichever practical you complete, you must make sure you know the
procedure for the other.
Option 1: Using brine shrimp
Many organisms’ ability to reproduce is dependent on the environmental temperature. C
Brine shrimp cysts will hatch when kept in salt water at a suitable concentration and
temperature.
- -

1. Take five beakers and use a waterproof marker to label them with the temperatures
15 °C, 20 °C, 25 °C, 30 °C and 35 °C.

© Pearson Education Ltd 2019. Copying permitted for purchasing institution only. This material is not copyright free.
Practical activities have been safety checked but not trialled. Users may need to adapt the risk assessment
information to local circumstances. This document may have been altered from the original. 2
 2. Create a salt water solution in each beaker by dissolving 2 g of sea salt in 100 cm3 of
dechlorinated water. Use a stirring rod to ensure all the salt dissolves.
3. Slightly dampen a piece of graph paper with some salt water. Pinch a small number of
brine shrimp eggs and carefully sprinkle them onto the damp graph paper. Count 40
eggs (you will need to use a magnifying glass) and use scissors to cut around the eggs
on the graph paper.
4. Place the cut piece of graph paper face down in the first beaker of salt water. The eggs
should fall off the paper into the water in approximately 2 minutes. Use forceps to
remove the paper.
5. Repeat steps 3 and 4 for each beaker.
6. Place each beaker in the correct refrigerator or water bath. Store the beakers for
24 hours.
7. Prepare another beaker of salt water by dissolving 2 g of salt in 100 cm3 of
dechlorinated water. This beaker will be used to count the brine shrimp.
8. To count the hatched brine shrimp, place a lamp or other bright light source next to
each beaker. Use a pipette to remove the brine shrimp that swim towards the light
source and place them in the beaker of salt water you prepared in step 7.
9. Record the number of hatched brine shrimp at each temperature in a suitable table.
10. Repeat counting every day for two further days.
Option 2: Using seedlings
1. Take five Petri dishes and use the waterproof marker to label them with the
temperatures 15 °C, 20 °C, 25 °C, 30 °C and 35 °C.
2. Mould some cotton wool so that the bottom of each Petri dish is completely covered
with a layer of cotton wool approximately 1 cm thick.
3. Use forceps to count 20 seeds for each Petri dish. Ensure the seeds are spread out
evenly around each Petri dish.
4. Use a measuring cylinder to measure and add 30 cm3 of water to each Petri dish.
5. Place each dish in the correct refrigerator or incubator.
6. After 24 hours, add another 20 cm3 of water to each dish. Check the dishes every 24
hours to ensure they do not dry out, adding water as needed. Ensure that the same
volume of water is added to all Petri dishes.
7. After 5 days, observe the seeds and record the number that have germinated at each
temperature in a suitable table.
Analysis of results
Plot a suitable graph of your results.
Learning tips
• When removing the graph paper from the beaker of salt water, inspect it carefully to
ensure no eggs remain.
• Brine shrimps are small and very delicate. They will need to be handled with care. Ask
your teacher where to place the hatched brine shrimp that have been counted.
Questions
1. Use your graph to predict the optimum temperature for brine shrimp hatching / seed
germination success.
2. Suggest one reason why 35 °C was chosen as the maximum temperature for this
investigation.
3. Suggest two reasons – apart from temperature – why some of the organisms you used
may not have developed.

© Pearson Education Ltd 2019. Copying permitted for purchasing institution only. This material is not copyright free.
Practical activities have been safety checked but not trialled. Users may need to adapt the risk assessment
information to local circumstances. This document may have been altered from the original. 3
 4. The surface temperature of the Earth is increasing. Use your data to suggest how this
organism may be affected if the average temperature of its habitat increases from
27 °C to 32 °C.
5. Describe one of the approaches that could be taken to ensure the ethically responsible
use of animals in this practical activity.
6. Briefly suggest how, when using brine shrimp for this experiment, it would be possible
to investigate the effect of salt concentration on the development of the organism.
Number of 7. Explain why, when using seeds for this experiment, the volume of water added to the

germinated
seeds must be controlled.
seeds
Exam-style questions
i 1. In an investigation, radish seeds were sown on moist filter paper soaked in 30 ml buffer
F ***

Tintston
solution. A range of buffer solutions was used to investigate the effect of pH on seed
germination. In total, 20 seeds were used in each dish. At 2-day intervals, 20 ml of
E buffer solution was added to each dish.
The results are shown in the table below.
8-
6- Number of seeds germinating after 7 days

E X
pH
Number of seeds
2 3 4 5 6 7 8 9

0 0 2 4 14 16 15 3
8 germinated

(a) Plot a suitable graph using the data shown in the table.
(3)
(b) State the independent variable and dependent variable in this investigation.
⑪H value Number of seeds geminated (2)
(c) As well as the volume of buffer solution used, state three other variables that
should be controlled in this investigation.The concentration of the salt solutions
The quantity of food administered to the brine shrimp (3)
Amount of water administered to the nedlings -

(d) A student concluded that pH 7 is the optimum pH for germination for this species of
plant. Evaluate this conclusion. The number of needs
germinated
at puF is the compared to other pl.
highest (3)
(e) Explain why there was no germination at pH 2 and 3.
It is too acidic so the required for gemination have
aenatured :
enzymes (4)
(f) Suggest reasons why there was not 100% germination at pH 7.
some seeds have
already spoiled or are dormaut- (3)
(g) Calculate the percentage of seeds that germinated at pH 7. Show your working.
-x185 982
=

(2)
2. A horticulturalist has been asked by a local council to provide salt-tolerant flowering
plants for planting near roadsides and beaches. Roadside verges can become salty
due to salt being added to roads in icy weather and sprayed by cars onto the verges.
Too much salt in soil can prevent many plants from taking up water properly.
Outline how the horticulturalist would investigate which plants to supply.
(8)

© Pearson Education Ltd 2019. Copying permitted for purchasing institution only. This material is not copyright free.
Practical activities have been safety checked but not trialled. Users may need to adapt the risk assessment
information to local circumstances. This document may have been altered from the original. 4