TIPS!

Be very clear about the difference between the terms:

Precise: A measurement with very little spread about the mean value. One with a lot of decimal
places.
Accurate: A measurement that is close to the true value.
Reliable: The results can be repeated. The reliability of data within a single investigation can be
improved by carrying out repeat measurements.
Valid: Measurements made are affected by a single independent variable only. They are not valid if
the investigation is flawed and control variables have been allowed to change.

When drawing a table to record data, be sure to include the original observed measurements that
would be recorded during the experiments. You might include additional columns for calculated
values such as a difference or percentage change.

One purpose of preliminary practical work is to determine appropriate conditions or values of the
controlled variables for the main data collection phase of the investigation. You should refer to
specific conditions and variables that are relevant to your investigation.

Hypotheses and statistical tests make use of specific and precise scientific language. Use the
correct terminology of significant difference (T test – when testing two different discrete data sets)
or significant correlation (Spearman’s Rank correlation test – when testing for correlation between
two variables) to formulate a good null hypothesis.

Make sure that you directly answer the question. If you are describing changes from the usual
course of events, be specific about the changes and describe their nature by using terms such as
more / fewer / greater / slower.

When describing a method the phrase 'record the results' is very vague and should be avoided.
Specify exactly what should be recorded: for example, in the case of habituation, record the time
taken for the snail to fully re-emerge from its shell.

An experiment is not valid if:

 X factor has not been taken into consideration e.g. health, age, gender, mass…
 Small sample size used
 Study only carried out on one occasion
 Study does not represent the whole population
 Oter factors may not have been taken into consideration
 Sample only taken at one time period
 Wide variability of data

Systematic errors in measurements of dependent variable can be reduced by:

 When measuring the colour of the solution make sure to use a suitable reference cuvette / solution
 Make sure you calibrate the apparatus
1. The Effect of Caffeine on Heart Rate

Why Daphnia?

 Daphnia are small and show the results of the  They are abundant so easy to get hold of / No
experiment quickly damage to environment when a few are
 They have simple nervous systems so are less removed from it
likely to feel pain  Transparent so easy to measure heartbeat

Variables to be controlled are:

 Temperature  Daphnia must be the same age / size
 Volume of solution  Daphnia must have the same health
 Time of acclimatisation  Counting time

Method:
Independent variable: Caffeine concentration
Dependent variable: Heart rate of Daphnia

1. Place a Daphnia in each of 5 different solutions (4 different concentrations of caffeine and one with
distilled water to act as a control)
2. Leave Daphnia for 5 minutes to acclimatise
3. Immobilize the Daphnia using a little cotton wool in a cavity slide and observe under microscope
4. Count and record the no. of heartbeats in one minute
5. Repeat 5 times at each concentration and allow means to be calculated

Outcome: As caffeine concentration increases, heart rate also increases

How does caffeine increase heart rate?

 Increasing caffeine concentration causes the electrical activity of the sinoatrial node to increase,
making it depolarize. As it depolarizes, the right and left atria contract and the impulse travels to the
atrioventricular node where, after a delay of about 0.13 seconds, the impulse continues to travel
towards the ventricles. This delay ensures that the atria have finished contracting and ventricles are
full. The signal then reaches the Purkyne fibres that conduct the impulses to the apex of the
ventricles where contraction begins and travels upwards towards the atria.
 Caffeine also affects the ventricles, leading to an increase in the rate of contraction and relaxation of
each heartbeat. This means that, as well as beating faster, the heart's individual beats are
associated with an increased cardiac output.

2. The vitamin C Content of Fruit Juice

Variables to be controlled:
 Mass of fruit / age / source / …  Volume / concentration of juice / DCPIP
 Time for storage  Temperature
 Method of juice extraction  Same end point colour

Method:
Independent variable: Fruit juice
Dependent variable: Volume of juice required to decolourise 1cm3 of DCPIP

1. Put 1cm3 of DCPIP solution into a test tube

2. Fill a plastic syringe with juice and add drops to the DCPIP until the blue of the DCPIP is lost.
3. Record the volume of juice added
4. Repeat 5 times for each juice to calculate means
5. To calculate the actual Vitamin C concentration, the DCPIP solution must be calibrated. A solution of
known Vitamin C concentration is added to 1cm3 of DCPIP until it is decolourised and the volume
recorded.
6. Conc. of Vitamin C in juice = (Con. of Vitamin C solution x Volume of Vitamin C solution needed to
decolourise 1cm3 DCPIP) ÷ Volume of fruit juice needed to decolourise 1cm3 DCPIP
Limitations:
 Difficulty in controlling temperature  Some loss of solution when transferring from
 End point difficult to judge as needs to be just one beaker to another
when blue colour disappears especially in  Accuracy of measuring equipment
highly coloured juices

Storing fruit at low temperatures slows down decay because:

 Low temperature reduces / prevents growth of microorganisms
 Low temperature reduces activity of enzymes
 Less kinetic energy means fewer collisions / fewer cell divisions

3. The effect of Temperature on Membranes

Factors that affect the permeability of the beetroot cell membrane are:
 Temperature  Duration  pH
 Age  Prior treatment with  Bile salt
 Storage solvent

Variables to be controlled are:

 Source / Species of beetroot  Time left in water or solution
 Age / Size of beetroot  pH
 Volume of water / solution used

Method:
Independent variable: Temperature of water
Dependent variable: % transmission of light through resulting solution

1. Using a cork borer and knife, cut 5 pieces of beetroot equal in mass and size
2. Rinse the beetroot pieces with water and gently pat them dry with tissue before using them as, when
cutting, pigment is released from broken cells and must be removed before starting or solutions will
be darker than they should
3. Place one piece into each of 5 tubes and add 5 x 1cm3 water to each one
4. Place each tube into a water bath of different temperature (e.g. 15, 20, 25, 30, 35)
5. Leave for 15 minutes
6. Remove beetroot and shake tubes to disperse dye.
7. Calibrate the colorimeter using distilled water in a cuvette as a reference / control.
8. Take readings of absorbance of the water in the tubes
9. Take 5 repeats at each temperature and calculate means

Results and reasons: As temperature increases, % transmission slightly increases. This is due to
membrane molecules gaining more heat energy and vibrating more, creating large gaps in the
membrane that enable dye to be released. Proteins in membrane may become denatured, leaving
large pores through which the dye leaks.

Limitations:
 Pigment is not equally distributed throughout  Some beetroot may have skin on affecting
the beetroot surface area
 Size of beetroot is difficult to control
4. The Effect of Enzyme Concentration on the Rate of Reaction

Variables to be controlled are:

 Temperature: use a water bath  Concentration of substrate
 Volume of enzyme  pH
 Volume of substrate

Method:
Independent variable: Concentration of enzyme
Dependent variable: Time taken for enzyme to break down substrate

1. Take 5 test tubes. In 4, place increasing volumes of trypsin solution e.g. (1, 2, 3, 4 cm3 ) and make
up the volume to 4 cm3 using distilled water. The other test tube should be filled with 4 cm3 of water
to act as a control.
2. Add 5 of milk powder (casein solution) as substrate and start the stopwatch
3. Measure the cloudiness of the solution over time using a colorimeter (every 30 secs for 10 minutes)
against water as a reference / control
4. Repeat at least 5 times at each concentration and calculate means

As concentration of enzyme increases, rate of reaction increases up to a point, where all enzyme has
metabolised all substrate immediately

Why do enzymes work better at higher temperatures?

At low temperatures the reaction is slow because the enzyme and substrate molecules don’t collide
very often and move slowly. Increasing temperature increases kinetic energy and so frequency of
collisions. The substrate binds to the enzyme’s active site more often thus increasing the rate of
reaction. After the optimum temperature bonds holding the 3D – shape of the enzyme together start
breaking so it loses its shape and the enzyme-substrate complex can no longer form. The enzyme is
denatured.

5. Observing Mitosis

Safety:
 Risk of injury to hands by sharp knife or mounted needle so wear thick gloves or cut away from body
 Acid is corrosive so wear gloves to reduce risk of injury and safety glasses to reduce risk of injury to
eyes
 Stain may stain clothes and skin so wear gloves and lab coat
 Glass coverslip may break and cut your fingers so wear gloves to protect hands

Method:
1. Cut the last 0.5 cm off the end of actively growing garlic or onion root tips
2. Treat with acid to soften tissue by breaking down the middle lamella so that the cells will separate
easily when squashed
3. Break up the tips gently using a mounted needle on a microscope slide
4. Add toluidine blue to stain the chromosomes, warming if needed to intensify the stain
5. Place a glass coverslip on top and squash gently
6. Observe under the microscope
6. Totipotency and Plant Tissue Culture

Safety: Bacteria could grow in agar

Variables to be controlled:
 Same age / size seeds  Time allowed to grow
 All seeds should come from the same plant  Atmospheric carbon dioxide concentration
 Temperature  Humidity
 Light Intensity

Method:
1. Use week-old mustard seedlings. Cut off the top 2cm (stem and leaves) and suspend in agar in a
test tube
2. Leave for a week and look for new roots / leaves forming
3. Cells at the bottom of the stem differentiate to become new roots which demonstrates pluripotency

7. The Tensile Strength of Plant Fibres

Safety:
 The fibre could enter and injure the eye when it snaps so wear safety glasses to protect eyes
 Place layers of cloth beneath the mass hanger to stop masses from falling onto the foot and injuring
it

Variables to be controlled:
 Length of fibre
 Size of each individual
mass
 Width / diameter / cross-
sectional area of fibre
 Fibres must come from  Same prior treatment
the same plant species  Temperature
 Same age  Humidity
 Same parent plant to
reduce genetic variation

Method:
1. Soak nettle plant stems in water for a week to soften the tissues and allow the fibres to be easily
extracted
2. Select adequate fibre (taking into account all variables) and attach one end to a clamp and stand
then progressively hang masses on the other end
3. Record the mass at which the fibre breaks
4. Repeat 5 times at each thickness to calculate a mean

Outcome: Wider fibres are stronger

8. Plant Mineral Deficiencies (Optimum concentration of substances

Safety:
 There are no significant safety issues Mineral / Plant / Enzyme allergies or irritants
 Hypodronics may provide good growing conditions for bacteria/fungi

Preliminary Work:
 See if proposed method will work
 See if the plant chosen will grow in
hydroponic unit
 Select a range of concentrations
 Check for suitable conditions for digestion
e.g. temperature, pH
 Find a suitable method of measuring growth
 Check for most suitable conditions for growth
of plants
 Select suitable time scale for measuring
growth / stain digestion
Things affecting enzyme action are: protein type, volume of solution, stirring, pH, temperature, surface
area, protein concentration…

Variables to be controlled:
 Volume of mineral solution  Light intensity  Genetically similar à Same
 Concentration of solution  Age / size of plant / seedling parent plant
 Species of plant  Temperature

Method:
Dependent variable: E.g. mass of plant tissue, mass of fruit, length of shoot, number / colour of
leaves. Description of method of measuring change in dependent variable
Independent variable: Concentration of magnesium. Range of suitable concentrations suggested (at
least 5) (0 (control), 10%, 30%, 50%, 70%, 90%, 100%)

1. Take six plants / seedlings and place each of them into a test tube with a different concentration of
solution (one with distilled water to act as a control)
2. Cover each tube with foil to exclude light and prevent algae growth that could affect concentration of
mineral ions
3. Leave the tubes for a week
4. Record changes in mass / height / root length
5. Repeat at each concentration / for each mineral ion 5 times and calculate mean
6. Use of graph to identify other values of concentration to test to identify optimum concentration

Recordings, Representation and Analysis:

 Table matching method with headings and units
 Change in growth calculated e.g. by measuring change in length
 Means calculated from repeat data
 Scatter / line graph format with correctly labelled axes
 Use of graph to estimate range for optimum / to identify other values of concentration to test to
identify optimum concentration
 Concentrations above those first reaching maximum rate of digestion would be wasteful

Limitations:
Difficult to control all variables affecting plant growth / protein digestion e.g. seeds do not germinate at
the same time, genetic differences between the plants… / surface area of stain, protein concentration

Limiting factor(s):
 Experimental conditions may not match those normally used
 More than one type of mineral for effective growth of plants
 Difficult to measure the dependent variable
9. The Antimicrobial Properties of Plants/Secretions

Ethical Issues:
 Welfare of frogs e.g. frogs should be kept in suitable conditions / not be harmed when collecting
secretions
 Return the frogs to their habitat after using them
 Avoid skin contact with frogs e.g. wear gloves when handling them / wash hands after handling them
/ wear eye protection
 Prevent the growth of harmful bacteria / exposure to harmful bacteria

Safety:
 Wipe working area with antiseptic solution / work close to a Bunsen Burner which sets up convection
currents of sterile air to prevent growth of unwanted harmful bacteria / contamination
 Secure lids with tape. Don’t seal completely to avoid pathogenic anaerobic bacterial growth
 Don’t use 37 ºC as this is human body temperature & could encourage pathogenic bacteria to grow

Preliminary Work:
 Practise proposed method / see if proposed method will work
 Allows selection of appropriate species of frog to be worked out
 Carry out experiments to determine a suitable method for collecting secretions from frog
 Carry out experiments to determine appropriate concentration / volume of frog secretion
 Carry out experiments to determine most appropriate method of applying the secretions to the plates
 Carry out experiments to determine best parameters for another named variable e.g. suitable
timescale for measuring inhibition of bacterial growth/conditions for growth of bacteria/type of
bacteria
 Determine best method of measuring dependent variable

Variables to be controlled:
 Concentration of plant  Same volume of plant material  Bacterial species (E. Coli)
material / antibiotic / antibiotic on each disc  Temperature
 Lawn of bacteria on petri dish  Disc size

Method:
Dependent variable: Zone of inhibition / absorbance of culture
Independent variable: secretions from different frogs / antimicrobial solution

1. Prepare, under sterile conditions, petri dishes with a thin layer of agar in them
2. Once the agar is set, spread a drop of bacterial culture (E. Coli) over the surface using a sterile
glass spreader to form a lawn
3. Prepare the extract by crushing material using a pestle and mortar with alcohol if necessary
4. Dip small disk of blotting paper and allow to dry
5. Minimally lifting the lid, place on the centre of the agar and press lightly
6. Secure lids with 2 pieces of cellotape but don’t seal completely in order to avoid pathogenic
anaerobic bacteria to grow
7. Incubate at 25 / 30 ºC for a week
8. Observe the plates and the zone of inhibition will be clear. Measure its diameter to give an idea of
relative antimicrobial strength / effectiveness against microbes
9. Repeats taken and means calculated

Recordings, presentation and analysis:

 Table which matches method described with  Graph type selected that matches the data to
headings and units be collected
 Change in bacterial growth calculated e.g.  Axes needs to be included
measure area of zone of inhibition /  Appropriate statistical test a t-test /
absorbance of culture Spearman’s Rank
 Means calculated from repeat data  Anomalous data ignored
Limitations:
 Difficult to control all variables (affecting bacterial growth)
 Other components of secretions may affect bacterial growth masking the effect of the antibiotics
 Difficult to standardise extraction of secretion
 Other variables related to frog e.g. age, size, gender
 Uneven spread of bacteria
 A variable may be acting as a limiting factor for bacterial growth (give example)
 Need to test effect on more than one type of bacteria

10. Study of the Ecology of a Habitat

Safety:
 Possible risk of coconuts falling so collect coconuts beforehand
 Possible risk from indigenous animals unidentified plants, insect bites…
 Possible irritant / allergenic material so wear gloves
 Working under sun so can be burnt. Wear protection against sunburn

Ethical:
 Minimise disturbance to the habitat  Change in food chain/food web

Preliminary Work:
 Practice proposed method to see if it will work
 Check for most suitable site/ time of the year
 Check for the most suitable method of measuring yield
 Select suitable intervals of sampling to give sufficient data for analysis
 Consider any other variables that need to be taken into account
 Consider impact of farming practices used like pesticides

Sampling Methods:
Random Sampling: Used when measuring density of a plant species or slow moving animals.
1. Set up grid using tape measure, use random numbers to generate points to place ± 10 quadrats
2. Count number of chosen species in each quadrat or estimate % abundance
3. Density = Total no. of plants counted / (Area of one quadrat x Total no. of quadrats taken)

Systematic Sampling: A line transect is used to study changes in plant species across an area.
1. A tape measure is laid along several zones to be looked
2. At least 10 quadrats are then placed at regular intervals
3. Record data

Variables to be controlled (Abiotic Factors):

 Light intensity  Temperature  O2 concentration
 Surrounding vegetation  Soil water  pH
 Slope  Humidity

Record, present and analyse:

 Clear table which matches method  Scatter/line/bar graph format with correctly
description with headings and units labelled axes
 Means calculated from repeat data  Use of correlation test (Spearman’s Rank, T
test)

Limitations:
 Difficult to control all variables (abiotic factors
affecting the variables being investigated)
 Difficult to standardise measurement of yield
 Difficult to harvest coconuts high on palm
 Storms will cause damage to palm trees close
to edge of sea
 Difficult sampling technique due to e.g. uneven
arrangement of palm trees
 Laboratory conditions may not relate to what  Movement of organisms
happens in reality/real life situation  Sampling taken within a small amount of time
 We assume that the species is evenly
distributed throughout the area and that the
placing of the quadrats is entirely random

11. Effect of Temperature / Conc. On Development of an Organism

Safety:
 Tissue culture may provide good growing conditions forbacteria
 Possibility of an allergic reaction to the plant growth regulators / plant material
 Use of sharp instruments

Preliminary Work:
 Practise proposed method to see if proposed
method will work
 Determine appropriate dependent variable
 Check most suitable conditions for growth of
plant tissue
 Select suitable timescale for measuring growth
rates
 Check for other variables that need to be taken
into account / controlled
 Check if type of plant / tissue used is affected
 Check for (suitable) range of concentrations of
plant growth regulator to be used

Variables to be controlled are:

 Same age / size of organism  O2 concentration  Food
 Same parent organism  Mineral Concentration  Time
 Light intensity  Water  pH

Method:
Dependent variable: e.g. % change in mass of plant tissue / no. of hatched shrimp / height of seedlings
Independent variable: Conc. of plant growth regulator / temperature (at least 5 different ones)

1. Take at least 5 repeats and calculate means

2. Maybe give a control if appropriate
3. Specific descriptions of plant tissue culture provided (e.g. need to grow on nutrient gel, aseptic
conditions, antibiotics in gel to prevent growth of microorganisms…) Same for Totipotency and Plant
Tissue Culture
4. Consideration of time period over which the growth will be measured

Limitations:
 Difficult to control all variables affecting tissue
growth + example e.g. exposure to bacteria
 Damage to plant tissue during preparation may
affect growth
 Other limiting factor for plant tissue growth
 Need for more than one type of plant growth
regulator for effective growth

Any difficulties in the method you have proposed?

An increase in environmental temp causes the yield of some crops to decrease because less material
is stored. As temp increases, so does respiration/photosynthesis/metabolism which causes increase in
GPP. Enzymes are more effective at higher temperatures below an optimum. Temp has a greater effect
on respiration than photosynthesis so NPP will be smaller as NPP = photosynthesis – respiration.
Photosynthesis may be limited by another factor.
12. DNA Amplification Using PCR

Method:
A mixture is prepared containing: the DNA sample, DNA polymerases (with high optimal temperatures),
DNA primers (short, single-stranded lengths of DNA that are complementary to those at start of STRs,
have fluorescent markers attached that aid the production of the final profile) and nucleotides
The mixture is the placed into a PCR machine where it undergoes the following cycle:
1. Sample is heated to 95 ºC. This separates the double helix into two strands.
2. Mixture is cooled to 55 ºC. Allows the primers to bind to the start of the STRs.
3. Further heating to 70 ºC. DNA polymerases attach to the primers and extend them, replicating the
STR sequence and the adjacent DNA.
4. Cycle is repeated for about 25-30 times, which takes about 3 hours, to produce a mixture of
different-length fragments unique to the individual.

The properties of the enzyme relevant to its biological activity in the amplification process:
 Enzyme is heat stable
 Its optimum temperature is 70-80 ºC
 It synthesises a new strand of DNA complementary to the template strand in one direction. A primer
is needed to begin synthesis of the complementary strand

Effects of temperature on production of DNA:

 In context of denaturation of DNA / 90 to 95 ºC / step 1; If temp too low DNA strands won’t separate
 In context of primer annealing / 40 to 70 ºC / step 2; If temp too high < binding of primers will occur
 In the context of extension / 70 to 80 ºC / step 3;If temp too low synthesis of new DNA strands would
not be completed. If temperature is higher than 95 ºC, the enzyme will denature.

Collect and analyse samples from more than one individual of each species because:
 One individual does not represent the whole species
 There will be genetic variation between individuals of the same species, testing more than one
sample will control for these differences
 Improves reliability of the data

Examine the DNA of 5 different genes because:

 Some genes might have little or no variation
 It will allow scientists to determine how closely they are related

13. Gel Electrophoresis

After PCR, gel electrophoresis is used to separate fragments by size and a band pattern is produced

Method:
1. Separating the fragments
2. Sample mixture mixed with coloured dye and placed carefully into wells at one end of agarose gel.
3. The gel is immersed in a buffer solution in a tank and a potential difference is set up across it.
4. DNA is –vely charged so will move towards the +ve electrode at the other end of the gel.
5. Smaller fragments will move faster so mixture is separated out into a pattern of bands.
6. The wells have a reference mixture of DNA fragments of known length to compare your samples to.

Viewing and comparing the fragments:

1. Fluorescent primers glow under UV light and allow a gel photo to be taken
2. DNA can be transferred from the gel to a nylon membrane by Southern blotting, which can be
treated with a DNA probe. This binds to the bands and carries either a fluorescent or radioactive
marker. Radioactive ones can be seen using autoradiography
3. Coloured DNA probes can be added to gels to see the bands directly

Graphs can be plotted of size of fragment against level of fluorescence (gives abundance of fragment)
14. Effect of Different Antibiotics on Bacteria

Same as Antimicrobial Properties of Plants

15. Investigating Respiration

Method:
Place known mass of organism (maggots) into the respirometer.
Allow time for them to acclimatise to their surroundings and then move the drop of coloured liquid back
to 0 on the scale using a syringe.
Start the stopwatch and note the position of the coloured liquid at regular intervals of 5 minutes.
Subtract the final value from the first to give the overall distance moved.
Usevolume of oxygen uptake = π r2l (l = distance moved by liquid in tube).
The volume divided by time will give the rate of respiration.

Variables affecting respiration are:

 Type/source of seeds  No. of organisms  Equilibration / acclimatisation
 Mass/number of seeds  Temperature of water baths e.g. Time left before
 Age of seeds between 20-40ºC measuring
 pH  Amount of soda lime  Moisture / humidity / water
 Time

Yeast will respire faster using glucose because glucose is the starting point for glycolysis reactions in
respiration; it is the first molecule to be phosphorylated. / Yeast will respire sucrose faster because it
can be broken down into molecules of glucose and fructose; providing double the substrate for
glycolysis / Yeast will respire sucrose more slowly because sucrose needs to be hydrolysed to glucose
and fructose in order to be used in glycolysis. / Rate of uptake of sugars differs: larger molecules may
be taken up more slowly.

Effects of:
 No oxygen during investigation = No/less movement of the liquid in the respirometer. No/less
change in volume/pressure of the gas. Aerobic respiration stops / Anaerobic respiration takes place.
Anaerobic respiration produces no carbon dioxide.
 Increasing temperature = Will increase rate of respiration (as it is enzyme controlled) but also the
volume of air in the apparatus
 Increasing air pressure = Will reduce the volume of air in apparatus

16. Effects of Exercise on Tidal Volume and Breathing Rate

Method: This uses a spirometer. Adding air to the chamber makes the lid of the chamber rise in the
water, and removing air makes it fall. Movements of the chamber are recorded using a kymograph (pen
writing on a rotating drum). The volume of air the person inhales and exhales can be calculated from
the distance the lid moves.
1. The apparatus can be calibrated so that the movement of the lid corresponds to a given volume.
2. The chart recorder can be set to move at a known speed.
3. A canister containing soda lime is inserted between the mouthpiece and the floating chamber. This
absorbs the CO2 that the subject exhales.
4. A disinfected mouthpiece is attached to the tube, with the tap positioned so that the mouthpiece is
connected to the outside air. The subject to be tested puts a nose clip on (to ensure no breathing
occurs via the nose), places the mouthpiece in their mouth and breathes the outside air until they
are comfortable with breathing through the tube.
5. Switch on the recording apparatus and at the end of an exhaled breath turn the tap so that the
mouthpiece is connected to the spirometer chamber. The trace will move down as the person
breathes in. After breathing normally the subject should take as deep a breath as possible and then
exhale as much air as possible before returning to normal breathing.
6. Repeats could be for same student at same time each day for a week or with 10 different students
(same age, gender, health, etc.)

Variables to be controlled:
 Same person / age / gender / time of day  Standardise exercise
 Temperature  Breathing must be measured over a set time
 Diet before testing… (e.g. 5 minutes)

How breathing is controlled by the nervous system in response to changing positions e.g. standing up
and sitting down: More energy is needed when standing up. The sympathetic nerve increases heart
rate. The ventilation centre in the medulla responds to chemoreceptors in the carotid that detect
changes in levels of carbon dioxide in blood. Motor cortex. Nerve impulses go to muscles involved in
breathing.

17. Investigating Habituation to a Stimulus

Ethical issues:
 Snails must be handled carefully so as not to  Snails should be released into the wild after use
harm or stress it

Safety:
Snail secretions may irritate skin or cause allergies or carry microbes à hands should be washed
thoroughly before and after handling snails

Variables that affect it are:

 Temperature  Light Intensity  Gender
 Background Noise  Species
 Humidity  Age
Method:
Dependent variable: Time taken to fully emerge
Independent variable: Number of pokes

1. Place snail on clean, firm surface

2. Allow time until snail has fully emerged from shell and has acclimatised
3. With a moistened cotton wool bud, firmly but carefully touch the snail between the eye stalks,
starting the stopwatch immediately
4. Record the time taken for the snail to fully re-emerge
5. Repeat the touch and timing for a total of 10 touches

Outcome: As the number of stimuli increase, the time taken for the snail to re-emerge decreases.

Limitations:
 Snails already handled before the experiment may not react in the same way
 Determining when a snail has fully emerged
 Lack of moisture may encourage snail to stay more in its shell

An increase in temperature increases the rate of chemical reactions in the snail, allowing it to re-
emerge more quickly. To control temperature use incubator/conditioner at constant temperature
suitable for snail

Calcium ion involvement in habituation: Repeated stimulation affects calcium channels. Fewer calcium
ions enter the pre-synaptic membrane and so less neurotransmitter is released into the synaptic cleft.
Less depolarisation of the post-synaptic membrane will occur and fewer sodium channels will open. No
action potential will be generated and so no impulse sent, no response is observed.