Biology A2 Snab Coursework

GCE
Edexcel Advanced Subsidiary GCE in

Biology (Salters-Nuffield) (8048)
First examination 2006
Edexcel Advanced GCE in

Biology (Salters-Nuffield) (9048)
First examination 2007
February 2006
February 2006
Biology (Salters-Nuffield) (8048/9048)
Edexcel Advanced Subsidiary/Advanced GCE

in
Coursework guide
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Acknowledgements
This specification has been produced by Edexcel on the basis of consultation with
teachers, examiners, consultants and other interested parties. Edexcel acknowledges
its indebtedness to all those who contributed their time and expertise to the
development of Advanced Subsidiary/Advanced GCE specifications.
Authorised by Jim Dobson

Prepared by Kath Skillern
Publications code UA016953
All the material in this publication is copyright
Edexcel Limited 2006
Contents
Introduction
Visit or issue report (Unit 6133/01)
Introduction
Exemplars
Identifying and describing two aspects of biology
Account of biological principles
Communication
16
Student checklist (Unit 6133/01)
21
Practical work review paper (Unit 6133/02)
23
Safety and use of apparatus
24
Producing reliable and valid results
24
Significance of results/data
24
Presenting data
24
Using results to draw conclusions
25
Final advice
34
Choosing the best practical
34
Coursework investigation (Unit 6135/02)
37
Assessment criteria Research and rationale
37
Exemplar 1 Jennys work
38
Exemplar 2 Michaels work
40
Assessment criteria Planning
42
Exemplar 3 Samirs work
43
Exemplar 4 Robyns work
47
Assessment criteria Implementing
51
Exemplar 5 Marcos work
52
Exemplar 6 Shazias work
55
Assessment criteria Observing and recording
60
Exemplar 7 Susans work
61
Exemplar 8 Gabriels work
65
Assessment criteria Interpreting and evaluating
68
Exemplar 9 Jatinders work
69
Exemplar 10 Davids work
75
Assessment criteria Communicating
78
Exemplar 11 Rosannas work
79
Exemplar 12 Jans work
95
Exemplar 13 (maximum mark) Katharinas work
Student checklist for project skills (Unit 6135/02)
104
38
Introduction
Edexcel is pleased to offer this coursework guide to support teachers in their work
with students for the assessment of the AS units 6133/01, 6133/02 and the A2 unit
6135/02.
Examples of students work have been obtained from the pilot cohort September
2002 to June 2004. The assessment is based on the criteria published in the Advanced
Subsidiary and Advanced GCE in Biology (Salters-Nuffield) 8048/9048 specification.
The examples used in this guide have been assessed by Edexcels senior examiners.
Edexcel wishes to thank the schools, teachers and students who participated in the
production of the exemplar material.
All of the examples used in this coursework guide have been made anonymous and
any names given are fictional.
In addition to this coursework guide there will be further support from a consultancy
service and a series of professional development meetings. For more information
please contact Edexcel on 0870 240 9800 or visit our website at www.edexcel.org.uk
or the Heinemann page at www.advancedbiology.org.
UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

Issue 1 February 2006

Visit or issue report (Unit 6133/01)
Introduction
Those schools involved in the pilot of these qualifications (20022005) should note
that these new specifications are slightly different for Section B.
One strand, (a), still deals with the biology involved for the visit or issue and should
be straightforward.
However, another strand, (b), now looks at future developments and also refers to
uses which can be taken as methods employed by biologists. Students can either
look entirely at future developments, entirely at uses (meaning methodology) or
at a combination of the two as long as the future developments are clearly linked
to the uses described.
The third strand, (c), now requires students to evaluate critically the validity of the
information gained whether from websites, books or the person(s) conducting the
visit.
Note: quotes from students work in bold italic are only a small percentage of the
actual work submitted and are for illustrative purposes only.
Exemplars
We have provided five examples (A-E) of student work that the moderator has
commented upon. Quotes from the students' work are highlighted in bold. The
moderator has identified where and why marks have been awarded in relation to the
assessment guidance, and has offered a final commentary with suggestions for
improvement.
Exemplar A A visit to Batemans Brewery.
Exemplar B A visit to Cranfield University for prospective A-level students to give
an insight into biology as a career.
Exemplar C Deforestation: the global assault continues.
Exemplar D Information leaflet on glaucoma.
Exemplar E Biology issue report.

Identifying and describing two aspects of biology

A Identifying and describing two aspects of biology
a
The purpose or significance of two aspects of biology are described

within the context of the chosen visit or issue.
There is some appreciation of the ethical, social, economic or

environmental implications of the biology encountered within the
context of the visit or issue.
The significance of both aspects of biology are described clearly

within the context of the chosen visit or issue.
There is a clear explanation of the ethical, social, economic or

environmental implications of the biology encountered within the
context of the visit or issue.
Total marks
2 marks
4 marks
Exemplar A A visit to Batemans Brewery

Student A produced one of the best visit or issue reports in the first two years of
the GCE in Biology (Salters-Nuffield) pilot, gaining the maximum mark of 20. The two
aspects of biology they connect were The Raw Materials for Brewing and
Fermentation. On the first page of student As report they not only identified both
of these but also indicated on which pages they were to be found.
Target Audience AS/A-level biology students
First Biological Principle The Raw Materials for Brewing (pages 2-4)
Second (major) Biological Principle Fermentation (pages 5-8)
Word Count 1435 words (not including Bibliography or Figures 1-4). Note
Figure 1 contains 65 words.
Student A then went on to give quite a detailed account of malt (and how it is
produced), water, hops and finings for their discussion of The Raw Materials for
Brewing.
The embryo must be small (ie low nitrogen content) so that most of the ear is
starch and large amounts of sugar can be extracted for fermentation.
Water composes 90% of the beer and therefore substantially affects flavour.
Hops are added in the copper, where the rolling boil physically breaks down
the hops releasing the oil (containing the bitter flavour compounds lupulin and
humulone) into solution.
Isinglass finings (collagen from the swim bladder of fish) are added during
conditioning.

The main aspect for student A was Fermentation. They gave a very detailed
account of:
the importance of glycolysis, with diagrams and details of the biochemistry

involved
the three strains of yeast used and how infections by wild yeast or bacteria were
prevented
the importance of temperature for fermentation, ie warmer temperatures mean

more unpleasant by-products such as esters and diketones
specific gravity changes
secondary fermentation and other uses of fermentation.

Fermentation takes place in large vessels at a controlled temperature (lower
temperatures produce fewer by-products, warmer more eg esters and
diketones, which have low flavour thresholds ie humans can taste them at
very low concentrations therefore if concentrations change, sales may be
affected) and pH (optimum conditions).
The yeast is checked every week for infection under the microscope.
These details meant student A gained two marks for strand (a).
Student A also gained two marks for strand (b). Ergot contamination was
discovered, meaning the malt had to be checked to ensure it was safe for human
consumption. The environmental advantages of using organic grain for malt, ie no
chemical fertiliser or pesticides, were also considered.
Environmentally speaking, this was good (no added fertilisers or pesticides)
but organic grain is inconsistent with varying size, nitrogen and starch
content.
The economics of using organic grain, which is more expensive, and the use of
isomerised hops were also considered. They both improve the consistency and
quality of bitterness but are more expensive for the small brewery.
For Fermentation, student A showed how excess yeast is sold to farms or the local
Marmite factory, ie disposed of in an environmentally friendly way, with economic
benefits for the brewery:
Yeast is reused at the brewery. Excess yeast is sold to farms or the nearby
Marmite factory ie yeast is disposed in an environmentally way and fewer of
the earths resource are used.
Student A also talked of mechanical carbonation of the beer being too expensive
for the small brewery.
Total: 4/4

Moderators comments
Maximum marks were gained for strand (a) because both aspects were discussed
and analysed in some detail. The importance of both for the successful brewing
of beer was also made clear throughout the report.
Maximum marks were gained for (b) because at least two economic,
environmental, social or ethical considerations were looked at for both aspects.
Exemplar B A visit to Cranfield University for prospective Alevel students to give an insight into biology as a career
Student B identifies the Significance of Two Aspects of Biology: Animal Testing
and Tumour Marking on page 2 of their work, but then on page 4, they identify two
more biological principles as Polymerase Chain Reaction and Gel Electrophoresis
there was no hint in the title and it was difficult to work out what the report was
concentrating on.
For strand (b), there was some discussion of ethics in that the Christian view of
animal experimentation was briefly discussed and there was some mention of the
cost of research projects:
Testing on animals for cosmetic purposes takes place for products like
shampoo using the draize test. Many people believe that animals should not
be used in this way for commercial purposes.
However, neither of these was actually tied to the two principles highlighted, namely
Polymerase Chain Reaction or Gel Electrophoresis. Consequently, strand (b)
gained zero marks.
Total: 1/4
Moderators comments
Student B gained only one mark for strand (a) because four principles were
described yet only Gel Electrophoresis was dealt with in any great detail,
showing how it was relevant to the work of Cranfield.
Strand (b) should have had at least two of social, economic, environmental or
ethical issues discussed for each of the two biological principles. In this case, it
was impossible to work out which two aspects the student was concentrating on
since four were unidentified.
In order to gain higher marks the student would need to:
make the two principles more obvious, preferably on page 1 in the title
make sure that both principles are discussed in reasonable detail, and also make
sure that it is obvious how important both are to the work undertaken at
Cranfield
discuss at least two of social, economic, environmental or ethical issues for each
of the two biological principles. The ethics of Gel Electrophoresis or
Polymerase Chain Reaction themselves are hardly an issue, but the costs might
be, as would the social implications of treating cancer successfully.

Exemplar C Deforestation: the global assault continues

Although student C uses subtitles such as Environmental Impacts or The Economic
Struggle, they also used other subtitles which made it impossible to identify the two
main biological principles under investigation. Consequently strand (a) scored zero
marks.
However, strand (b) scored two marks because, although the two aspects were not
identified clearly, there was a lot of discussion on the environmental impact,
economic effect and also many social aspects such as urban migration and
employment generation.
Another extremely concerning deforestation impact is genetic erosion. This is
the loss of genetic diversity within a species caused by human activities and
environmental changes.
Faced with political decisions about urban migration, food production,
employment generation, national security, economic structural adjustment
and all the other issues that demand their attention, many governments have
opted to ignore deforestation.
Total: 2/4
Moderators comments
Student C did not clearly identify the two aspects of biology under
investigation.
identify clearly the two aspects of biology being considered and then discuss at
least two of ethical, social, economic or environmental implications for each
aspect.
Exemplar D Information leaflet on glaucoma

Student D produced an information leaflet on glaucoma and how cannabis could be
used in its treatment. Two marks were awarded for strand (a) because glaucoma and
cannabis were described as the two biological aspects to be covered in the leaflet.
This is an informative leaflet that explains the problems associated with
glaucoma, and explains how the use of cannabis can help.
Also, both aspects were dealt with in reasonable detail and the significance of each
was spelt out. Only one mark was awarded for strand (b) because, although both the
ethics and social advantages of using cannabis were discussed, there was no mention
of the social or economic issues associated with untreated eye diseases such as
glaucoma.
Total: 3/4

Moderators comments
Student D clearly identifies the two aspects of biology under investigation.
In order to gain a higher mark the student would need to:
mention that untreated diseases, such as glaucoma, could result in premature

blindness and remove a significant number of people from the workforce. This
would then cost a lot in benefits and other taxpayers would have to support
them. In addition, the social disadvantages of having more people dependent on
others could have been looked at.
Exemplar E Biology issue report

Student Es report clearly identified the two aspects of biology under investigation
and so scored two marks.
The purpose of this report is to investigate links between one form of cancer
treatment and its associated influence on female infertility.
Only one mark was awarded for strand (b) because there was a discussion of the
social implications and a reference to ethics but only for the idea of freezing ovarian
tissue.
Opponents to this form of treatment may hold the view that it is interfering
with nature
Total: 3/4
Moderators comments
Both aspects are identified and the relevance of both of these to female fertility
is discussed.
There is nothing on the economics of the treatments.
expand on the ethical discussion of the treatment and also include a look at the
social implications for the individuals involved.


B
There is a description of the biology relevant to their chosen aspect.
There is a reference to future developments or uses in this area of

biology.
There is reference to information or arguments obtained from three

or more sources.
There is a clear description of the biology relevant to their chosen

aspect.
There is some speculation as to future developments or uses in this

area of biology.
There is reference to the validity of the information or arguments

obtained from three or more sources.
There is a discussion showing some understanding of the biology

relevant to their chosen aspect.
There is informed speculation as to future developments or uses of

biology in this area of biology.
There is some evaluation of the validity of the information or

arguments obtained from three or more sources.
There is a clear discussion showing a thorough understanding of the

biology relevant to their chosen aspect.
There is discussion of possible implications of future developments or

uses in this area of biology.
There is a clear evaluation of the relative validity of the information

or arguments obtained from three or more sources.

Total
marks
3 marks
6 marks
9 marks
12 marks

Student A gained four marks for strands (a) and (b) because they not only identified
their main aspect as Fermentation on the first page, but went on to discuss a great
deal more biological detail such as:
well set-out equations and discussion of glycolysis showing how ethanol is

produced from glucose
a good discussion of the three strains of top-fermenting yeast used.

Batemans uses 3 strains (mixed) of Saccharomyces cerevisiae (topfermenting) yeast, the advantage of this being that the yeast collects on the
top of the vessel, protecting the alcohol from oxygen in the air therefore
preventing oxidation of the alcohol to acetic acid or vinegar.
The strain of yeast used must remain consistent resulting in consistent
flavours.
the importance of temperature control for flavours
the importance of the original specific gravity for final alcohol concentration
the use of secondary fermentation to carbonate the beer
other uses of fermentation to produce cheese, yoghurts, antibiotics or metal

extraction from low-grade ores.
Student A also gained four marks for strand (b), future developments or uses.
For future developments, they pointed out that:
The yeast genome was sequenced in 1996, therefore genetic engineering
may be used to create new strains of yeast with improved characteristics to
increase the efficiency of fermentation and the flavour of the beer.
Student A speculated that this might also mean a decline in continuous
fermentation and the use of higher gravity worts.
The term uses clearly refers to methods, ie what biologists actually do. Student A
linked their discussion of biological principles to what actually happens at the
brewery. Student A showed how the brewery used three strains of yeast and
highlighted the advantage of top-fermentation. In addition, student A discussed the
importance of controlling the temperature and how this is done, how the specific
gravity falls and how the brewery uses a secondary fermentation in order to
carbonate the beer.
Batemans only produces cask-conditioned beer ie beers that undergo a
secondary fermentation whilst in a sealed container.
For strand (c), student A referred to her bibliography at regular intervals. For
example, student A discussed how carbonation from a secondary fermentation is an
advantage for a small brewery because buying in carbon dioxide is too expensive
www.brewing.com/beer-carbonation/carbonation-methods.htm. Student A gained
four marks for strands (a) and (b).
10

In addition, as well as showing how the deliberate lowering of the temperature to

9oC once the specific gravity has reached 1.010o helps to keep some glucose available
for the secondary fermentation, student A compared the fall in specific gravity with
that of another brewery.
Also data from another brewery (Highwood) is similar, with specific gravity
falling from 1.045o to 1.009o overall.
Student A also used reference material to question the future use of continuous
fermentation as a result of genetic engineering of yeast,
www.mbaa.com/TechQuarterly/Abstracts/1977/tq77ab33.htm.
Total: 12/12
Moderators comments
Maximum marks were obtained because student A not only provided a great
deal of biological information about glycolysis and fermentation but also
speculated about the implications of genetic engineering for the future of
brewing.
Student A also looked at the methods actually employed at the brewery and
critically used her reference material either to make a point or to compare the
claims of one brewery with those of another.
In his report on the visit to Cranfield University, student B did not state the main
biological principle they were investigating, but it was taken to be Gel
Electrophoresis. Student B gained two marks for strand (a) because, although
there was quite a lot of biological detail in the report as a whole, only a small
amount was actually relevant to Gel Electrophoresis, ie structure of DNA or size of
DNA fragments after cutting with restriction enzymes.
Gel Electrophoresis is used for the identification of particular DNA molecules
by the band patterns they yield after being cut with various restriction
enzymes.
For strand (b), there was no mention of future developments for Gel
Electrophoresis or PCR but there was a brief discussion of how Gel Electrophoresis
was carried out, which gained one mark.
An agarose tank is prepared, acting as a support for separation of the
fragments of DNA. Holes in the gel hold the DNA solution.
There was a brief look to the future for alternatives to toxicology, but this was not
identified as the main biological principle and so could not be credited with a mark.
For strand (c) there were five references but none were mentioned in the text.
Acknowledgement would have gained an extra mark for strand (b) but, in this case,
there was no critical evaluation.
Total: 3/12

11
Moderators comments
The discussion of Section B was too descriptive.
The main biological principle was not identified and a considerable amount of the
biological detail involved was not relevant to Gel Electrophoresis, the principle
chosen.
There was no look at the future, and methods or uses were described only
briefly.
Three or more references/sources/materials were not analysed critically.
identify the main biological principle of the visit or issue
give far more biological detail relevant to the main biological principle. In fact, it
might have been better to have chosen Polymerase Chain Reaction as the main
principle for discussion
discuss future developments or improvements relating to Gel Electrophoresis,

or the methods involved, with more detailed analysis
look at far more critically at least three references or sources. For example, the
Cranfield University information pack is mentioned, but there is no questioning of
the claim that a safe effective drug results from the clinical trials? A cancer
research website was listed and, although examples of tumour markers were
given, there was no indication of how effective these were or indeed how they
worked.
Exemplar C
Deforestation: the global assault continues
The main principle under investigation was not identified by student C, but in the
deforestation report there was some discussion of:
drought and soil erosion
climate change
loss of biodiversity
loss of plants as a potential for medicines
genetic erosion
poverty.
The antiseptic phrase loss of biodiversity masks the fact that the annual
destruction of millions of hectares of tropical forests means the extinction of
thousands of species and varieties of plants and animals
However, the discussion was brief and somewhat descriptive, with insufficient
explanation of biological treatment, and scored only two marks for strand (a). Strand
(b) scored zero marks because there was no discussion of what biologists were
actually doing about deforestation and there were only a couple of lines about the
future, with a brief mention of the World Wide Fund for Nature.
Environmental organisations like WWF and Friends of the Earth are tying to
help save forests by campaigning for their urgent protection.
12

Strand (c) also scored zero marks because, although there were several reference
sources, none were used in the text and there was no attempt to use this reference
material to check on the validity of the information used. The report, as a whole,
was descriptive rather than analytical.
Total: 2/12
Moderators comments
The report was actually far too short, being only about 1000 words.
use the full 2500 word limit. This would allow far more detailed discussion,
especially of the biology involved. This could have been a more detailed look at
climate change or the importance of the forest habitat to species such as the
orang-utan, eg what tree species are most important
clearly identify the main biological principle under investigation
speculate on the future developments associated with deforestation, eg

reforestation or a more detailed look at what WWF or Friends of the Earth are
actually doing. In addition, look at methods used by biologists. If biodiversity is
the main aspect, then it would be reasonable to include reference to seed banks
or zoos as a way of preserving some species
use the reference material, ie both refer to it and examine it critically. For
example, a website reference to the World Resource Institute is given as
www.wri.org but there is no discussion of what this organisation is doing. Another
is www.rcfa-cfan.org/english/info.tree.html and includes details of how the
Canada Fund for Africa supports forestry and agroforestry research in Africa. Yet
this is not mentioned in the text or commented on.

This information leaflet on glaucoma gained only two marks for strand (a). Firstly,
student D did not identify the main principle that this leaflet was describing. Since
the details of glaucoma were rather descriptive, the use of cannabis was taken to be
the main principle. There was a brief discussion of the chemistry of THC (tetra-hydro
cannabinol) together with some of the medical problems that it could be used to
treat, eg as an antinausea drug or as treatment for glaucoma.
Once THC has entered the blood stream, it travels around the body until it
reaches the brain. Here it attaches itself to cannabinoid receptors and its
effect is felt.
Strand (b) only gained two marks because there was no mention of future
developments at all, but there was some discussion of how cannabis is used
medicinally.
If cannabis is smoked through a vaporiser, the THC is vaporised and nearly
all the carcinogens are left behind.

13
Strand (c) scored zero marks because there were only five reference sources, which
were impossible to follow up because of lack of detail, and in the text they were not
acknowledged or discussed critically.
Total: 4/12
Moderators comments
The discussion of Section B was too descriptive.
identify the main aspect clearly, in this case, how cannabis is used to treat
diseases such as glaucoma
provide a much more detailed biological background for cannabis itself, how it
affects the body and, also, how it alleviates some of the disease symptoms. For
example, student D does discuss how cannabis dilates blood vessels and explains
how this might help with glaucoma. However they could have expanded on this to
explain how this might help with the other diseases mentioned, ie asthma,
obesity
either give more examples of how cannabis could be used or speculate more
about its possible future use. For example, if it dilates blood vessels, could it
help with hypertension?
critically analyse some of the reference material and certainly make it possible
for the reader to follow it up. For example, the word proven is used for
cannabis easing the effects of glaucoma did only one reference suggest this or
several, and what sort of agreement was there?

The main principle was not identified by student E but strand (a) gained three
marks because there was a reasonable discussion of the biology associated with
ovarian tissue banking and the problem of cancer cells remaining in the tissue sample
taken.
Ovarian tissue contains follicles and immature eggs and the preserved tissue
can be transplanted back to the donor to allow the women to produce human
eggs.
Strand (b) gained four marks because there was quite a lot of detail on methodology
and, also, there was a comment on future developments.
Unfortunately further studies are required to determine the best option for
fertility treatment as there are safety concerns.
Whilst this experiment has not yet been successful on humans, a monkey in
Oregon has recently become pregnant following the treatment, and
researchers are confident that their findings show that there is potential for
normal ovarian function to be restored to cancer patients.
14

Strand (c) only just received one mark for the following comment. However, the
comment was not followed up.
The fertilised embryo developed to the four cell stage and was implanted into
her womb but she did not become pregnant. Despite this, researchers say the
findings prove the value of long-term tissue banking.
Total: 8/12
Moderators comments
There was a good discussion of the biology involved.
There was quite a detailed look at the methods used and possibilities for the
future.
There was a very brief look at the reference material used.
clearly identify the main principle being considered so that relevant discussion of
the biology involved can then be credited
be much more critical of the reference material being used, eg when discussing
the work of Kuluk Otkay on ovarian tissue banking
use research methods to try to find out who else is working on this and compare
their success claims or methods used.

15
Communication
C
Communication
The organisation and layout of the report shows evidence of some

planning for the intended audience, with use of some appropriate
graphs, tables, diagrams or photographs and technical terms, used
mostly correctly, where appropriate.
Spelling, punctuation and grammar are largely correct, all sources

used are acknowledged and not used excessively and the
presentation is mostly clear and logical. This is the maximum mark
available for reports which exceed 2000 words.
The organisation and layout of the report is well planned for the
target audience and is enhanced by carefully selected graphs,
tables, diagrams or photographs and correct use of appropriate
terminology where appropriate. Sub-headings of graphs, diagrams
and tables are appropriate and helpful.
Total marks
2 marks
4 marks
Spelling, punctuation and grammar are correct, all sources used are
fully acknowledged and used selectively and the presentation is
logical and concise. The report is within the word limit of 2000
words.

On the first page, where student A identified two aspects, they also noted the
audience to be AS/A-level students. Although not particularly original, they did write
reasonably simply and include lots of tables, pictures, graphs and diagrams. Their
impact was also broken up with subheadings in more of a magazine format and none
of the pages was just solid text. Student A gained two marks for strand (a).
In addition, for strand (b), the spelling, punctuation and grammar were excellent and
the report was concise, well-written and easy to read (1435 words). Not only did
student A include seven references to books or websites, they also acknowledged
these in the text. For example, when discussing clarification of beer, a reference was
given in the text using a footnote 2 for the second reference, in this case to
www.beer-brewing.com.
Total: 4/4
Moderators comments
The audience was identified and the write-up appropriately targeted.
Not only were several websites or book references given, but they were also
referred to individually in the text, ie it was easy to see where the information
had come from and it was easy to follow it up.
16

The visit by student B to Cranfield University was directed at prospective A-level
students and the report was reasonably well set-out with diagrams and subtitles.
This scored two marks for strand (a).
The report aimed at giving prospective A-level students an insight into
careers associated with biology.
However, although there were five references to websites, which gained one mark,
none were acknowledged or discussed in the text.
Total: 3/4
Moderators comments
See above.
ensure that references used were acknowledged and referred to in the text.
Exemplar C Deforestation: the global assault continues

The audience was identified clearly as GCSE students by student C, but the report
was not particularly well set-out. Only two pictures were used, with a few
subheadings and no graphs or diagrams to illustrate the point being made. It would
not have been a very interesting read for the intended audience, and so scored only
one mark for strand (a). Strand (b) also scored one mark because although there
were nine references to books or websites, none were acknowledged or referred to in
the text.
Total: 2/4
Moderators comments
See above.
make sure that the report is suitable for the intended audience. In this case,
more pictures or diagrams could have been used and more subheadings would
have broken up the text
acknowledge the references in the text. For example, a website address could
have been included for the photograph of the orang-utan representing the loss of
biodiversity.

17

The audience was identified by student D as older people who might be in an
opticians or doctors waiting room, but the leaflet was not that well-written or
clear. It is unlikely that older people would have found it easy to follow and
strand (a) scored only one mark. Strand (b) also scored one mark, because references
were difficult to follow up and were not acknowledged in the text.
Refer madness book.
Two pictures were used, but it was not obvious where they had come from.
Total: 2/4
Moderators comments
See above.
ensure that, if the audience is to be older people, the leaflet must be easier to
read and more attractively laid out with subheadings etc. Potential glaucoma
sufferers might also find the print size and font difficult to read
ensure references are clear and acknowledged in the text. For example, the two
diagrams could have had a website or book reference. Also, fewer than 1500
words were used and extra words could have been useful in giving greater detail
of the biology involved in cannabis action.

The audience was not identified clearly by student E.
It is a report intended for newspaper publication.
Not only was the target audience unclear, but the presentation of the report was
very dull. There were no subheadings or diagrams and only one photograph of the
doctor himself, which was not particularly informative. Consequently, strand (a)
scored zero marks. Strand (b) scored two marks, but only just. This was because
there were five website references, which were referred to.
With Otkays work, we appear to be much closer to the day when a young
woman diagnosed with cancer can both survive and subsequently have
children said Marian Damewood MD.
Total: 2/4
Moderators comments
18
See above.

be much more specific about the intended audience, ie tabloid newspaper or

broadsheet, general front page story or specialist article?
make sure that the style, presentation and readability of the article are suitable
for the audience
ensure that references are used and acknowledged in the text. In this case, by
simply putting the website address next to the relevant paragraph or using a
footnote.

19
20

Student checklist (Unit 6133/01)
Make sure that two aspects of biology are clearly identified and that the
relevance of both of these to the issue or visit is described.
Ensure that at least two of ethical, social, environmental or economic issues are
described fully for both of the biological aspects.
Clearly identify the main biological principle under investigation for Section B.
Thoroughly examine the biology involved for this main aspect.
Consider, in detail, the future developments for this aspect and/or the methods
used.
Comment critically on the information obtained from references given in the

bibliography.
Clearly identify the intended audience for the report and make sure that the
report is suitable for that audience.
Break up the text using relevant pictures, diagrams or subheadings so that the
report is a good read.
Ensure that several references are given and that they are acknowledged in the
text.

21
22

Practical work review paper (Unit 6133/02)

As an alternative to the GCSE style of practical coursework, the A/AS GCE in Biology
(Salters-Nuffield) specification offers the practical work review paper. The idea is to
allow students to enjoy and learn from their practical work, without having the worry
of constant assessment. Assessment is later on and is in a form which can build on
work done in the course.
Ten pieces of practical work are designated in the specification as core practicals.
1.1.17
Describe how the effect of caffeine on heart rate in Daphnia can be

investigated practically.
1.2.3
Describe how the effect of temperature on membrane structure can be

investigated practically.
1.2.11
Describe how enzyme concentrations and substrate concentrations can

affect the rates of reactions and how the effect of enzyme concentration on
reaction rate can be investigated practically.
1.2.17
Describe how Gel Electrophoresis can be used to separate DNA fragments of

different length.
2.3.6
Describe the stages of mitosis and how they can be observed practically.
2.3.13
Describe how the expression of a gene can be demonstrated practically by

induction of galactosidase.
2.4.7
Describe how to determine the strength of fibres.
2.4.9
Describe how to investigate the antibacterial properties of plants.
2.4.17
Discuss the possible relationship between CO2 levels and global warming and
how this can be investigated practically.
2.4.22
Describe how to investigate the effects of temperature on the development

of organisms (eg plant growth or brine shrimp hatch rates).
Strictly speaking, the definition of a core practical is one that can be used as the
basis for a question on the theory paper (Unit 1 or Unit 2). In practice, these 10
pieces of work, which have to be carried out, are likely to form the basis for work on
paper 6133/02. Also, students have to choose a piece of practical work to support
their answers to each of Questions 2 and 3 on this paper. This can be the same piece
of work but, again in practice, it is unlikely that this would be the best approach. For
Question 1, any practical work done in the course can be referred to.
Paper 6133/02 will assess students abilities to see elements within their practical
work that are common to most good science in this subject at this level. In that
sense, it builds on the work that most students will have done at GCSE level on the
nature of scientific enquiry. The specific requirements are on pages 2829 of the
specification (January 2005) and are reproduced here.

23
1 Safety and use of apparatus

Students should:
be able to comment critically on the suitability of apparatus for carrying out

specific practical tasks in order to achieve measurements appropriate to the
investigation
have an awareness of safety in the use of apparatus
during their practical work, consider the ethical issues arising from the use of
living organisms and for the environment.
This could be in the context of an evaluation of practical work they have taken part
in or when planning a new investigation.
2 Producing reliable and valid results

Students should:
appreciate that appropriate observations have to be made to validly test a

hypothesis or idea
appreciate that variables need to be identified, including both dependent and

independent variables, and, where possible, controlled or otherwise allowed for
be aware that valid results are derived through precise, repeatable

measurements or observations, made with apparatus and experimental
procedures that are suitable for the task
be aware that errors in readings can be systematic (values differing from the true
value by the same amount) or random (values lying equally above or below the
true value).
3 Significance of results/data
Students should realise that the significance of differences or trends within data is
dependent on the degree of error within the experiment.
4 Presenting data
Students should be able to discuss the most appropriate methods for presenting data
in order to identify trends and patterns clearly, and to an appropriate degree of
accuracy.
24

5 Using results to draw conclusions

Students should appreciate that statements, explaining trends and patterns in data
from an investigation, should be supported by evidence from the data and from their
own biological knowledge. The validity of a conclusion is dependent on the
experimental method used and the quality of the results obtained.
The review will require students to refer to the accounts they have made of the
practical activities carried out in Units 1 and 2. Students may have to include
examples of data tables, graphs or other relevant information in carrying out their
practical work review. They will have to submit to Edexcel the completed practical
work review and the complete accounts of two practical activities that have been
used as evidence.

25
Examples of questions, taken from June 2004 Unit 6133 Paper 02; these have
been used in the specimen papers.
Question 1
General principles of good science are tested in this question. Examples may be
asked for and can be taken from anywhere in the AS specification.
Topics covered to date (June 2005) include:
accuracy in measuring dependent variables
pre-treatment of experimental organisms
ethical issue of using organisms
reliability
types or error (as identified in the specification).
In answering this question students have, in the past, referred to a wide range of
practical work. Below is a list of the most popular ones:
heart rate of Daphnia
beetroot membrane
enzyme concentration
CO2 and temperature rise.
June 2004, Question 1:

(a) Describe what is meant by random error and give an example where your
experimental results were affected by random error.
(b) Distinguish between the accuracy and the reliability of results.
(c) Explain how you would ensure that any results you obtain are reliable.
Example 1 Part 1 (a):
Unfortunately the above answer does not gain any marks because it does not show
understanding of the meaning of the required term and, therefore, neither addresses
it nor gives a suitable example.
26

However, the answer below shows a clear understanding of the relevant term and
gives a good example.
Example 2 Part 1 (b):
The above answer does not attempt to define the terms asked for and, therefore, it
does not score any marks.
However, the answer below provides clear definitions for both terms and, therefore,
scores two marks.

27
Example 3 Part 1 (c):
The above answer struggles to score one mark, which has been awarded for the idea
that the experiment needs to be repeated many times. The student shows little
evidence of having understood what reliability means in this context.
However, in the answer below a clear understanding is shown.
Question 2
This requires a specific practical activity to be referred to and then used to answer
the questions. In this way, it does not differ from Question 3 and, therefore, both
questions are two of a kind.
Practicals used in the past to support this question (and Question 3) include:
heart rate of Daphnia
beetroot membrane
enzyme concentration
CO2 and temperature rise
antibacterial properties of plant extracts
strength of plant fibres.

From the practicals that you have completed during the course, select one suitable
practical report to support your answers to the following questions:
(a) explain what is meant by the following terms:
(i) Dependent variable.
(ii) Independent variable. (2 marks)
(b) identify the dependent variable in your report, and state how you ensured it was
measured to a suitable level of accuracy. (3 marks)
(c) describe how one other variable was or could have been controlled, and how it
may have affected your results. (2 marks)
28

The above is a full mark answer, but the student has spent a lot of time getting
there, largely because they have misjudged the amount of detail needed (only two
marks are available) and have not noticed that examples do not have to be quoted.
However, the answer below does not include anything useful and just quotes the two
terms at each other.

29
The answer on page 29 is a good answer, with reference to a suitable piece of

apparatus, a suitable variable and detail about its proper use. Students need to
constantly be looking at mark allocations for questions and parts of questions in
order to provide the necessary amount of detail. There are three marks available
here and it is clear what is wanted. One is obviously for correctly identifying the
dependent variable, which then leaves two marks to be gained. Just identifying the
apparatus used is not enough to gain both marks. In addition to identifying the
apparatus, students should be aware that accuracy is ensured by choosing suitable
equipment and using it properly, which will give students the two marks. Many failed
to get three marks for missing this last point.
The answer below fails to get any marks because of the fundamental mistake of
choosing the wrong variable as dependent. In one way this may seem harsh, but it
has to be remembered students have the paper for a week before they need to
answer the question.
Example 6 Part 2 (c):
The above answer achieves one mark for the fairly poor attempt to suggest the
effect that changes in temperature may have in an investigation where the
independent variable is caffeine. No attempt has been made to suggest how the
variable might be controlled. This could be a failure to read the question properly
and take account of what it required. It is a common error in this type of question
about confounding variables.
30

The above answer is clearly weak at most levels. However, laboratory air movements
might cause some problems. So the real difficulty here is not the rather poor choice
of variable but the lack of any attempt to specifically identify what it might do to
the data or how it might be controlled. Creating anomalies in my results or (in
other answers) affecting my results or similar vague comments will not achieve any
marks on their own. Something specific in relation to the chosen practical is always
going to be needed. This answer seems to be hinting that temperature might not
have been controlled as desired and so a temperature-related effect on, for
example, heart rate or rate of enzyme reaction could have been discussed.
Question 3
This question is similar in type to Question 2.
From the practicals that you have completed during the course, select another
suitable practical report to support your answers to the following questions:
(a) Explain two procedures or precautions that you took to ensure safe working. You
are reminded that these should specifically refer to the practical you have
chosen. (2 marks)
(b) Explain why the graph you have drawn is the most appropriate way of displaying
your results. (5 marks)

31

Relatively simple questions about safety can sometimes yield less than full marks.
Clearly, the practical chosen to support the answer can have a bearing on this. It
might be that the safety issues involved are few and not a great deal of explanation
is needed.
In this case, it might have been better to choose a different piece of work. It is
appreciated that any such choice will be a compromise. An answer such as the one
below suggests that the student was making up the safety points as they went along.
One just convinces the examiner, but the other one is judged to be too trivial. More
forethought might have led to a better discussion of ideas.
Any practical work involving the manipulation of bacteria is likely to yield a good
score in terms of safety questions, as in the answer below.
32


Scattergraph was crossed by the examiner in responses to this question. This is a
mistake in MS Excel, where line graphs are referred to as scattergraphs. Students
should be alerted to this.
Below is a full mark answer (as were many on this question).
Below is another full mark answer.

This student has made a reasonable, but vague, point about visual representation in
the first paragraph; however, the marking point (9) is gained later. The mark is given
for a line graph and this is often where the examiner has to check the appended
experimental report to make sure that this is true. After that, the student simply
repeats themselves and gains no further credit.

33
Final advice
It is important to review the relevant sections throughout the course. Students who
know exactly what they are to be examined on will do much better; they will avoid
making false statements and making things up.
Care should be taken when selecting practicals in light of the questions set in any
particular year. Guidance is given in the table on page 35 as to the appropriate
core practicals to choose.
Students must make good use of the week prior to sitting the paper to make
appropriate choices. Teachers can give generic advice, but must not, of course, tell
students what to do at any stage.
Choosing the best practical

Summary of the main assessable topics
A Choice of apparatus/method etc, to ensure appropriate/precise measurements.
B
Safety.
Ethics of experiments and observations.
D Reliability (aspect of validity).

E
Identification, control and/or measurement of independent variable and

dependent variable.
Identification, control and/or measurement of other variables.
G Error types.
H Dependency of significance of trends etc on degree of error.
I
Data presentation methods (graphs, tables, bar charts, etc).
Drawing conclusions from data.
34

Main assessable topics

Core practicals Units 1 and 2
1.1.17 Describe how the effect of caffeine

on heart rate in Daphnia can be
investigated practically
3?
1.2.3 Describe how the effect of

temperature on membrane structure can
be investigated practically
3?
1.2.11 Describe how enzyme

concentrations and substrate
concentrations can affect the rates of
reactions and how the effect of enzyme
concentration on reaction rate can be
investigated practically
1.2.17 Describe how Gel Electrophoresis

can be used to separate DNA fragments of
different length
3?
2.3.6 Describe the stages of mitosis and

how they can be observed practically
3?
3?
2.3.13 Describe how the expression of a

gene can be demonstrated practically by
induction of galactosidase
2.4.7 Describe how to determine the

strength of fibres
2.4.9 Describe how to investigate the

antibacterial properties of plants
3?
2.4.17 Discuss the possible relationship

between CO2 levels and global warming
and how this can be investigated
practically
3?
2.4.22 Describe how to investigate the

effects of temperature on the
development of organisms (eg plant
growth or brine shrimp hatch rates)
3?
Key:
3 = Yes
3? = Maybe
? = Probably not
= No

35
36

Coursework investigation (Unit 6135/02)
Assessment criteria Research and rationale

A Research and rationale
a
There is some attempt to provide a rationale for the choice of

investigation in terms of its scope and its relation to biological
principles.
Few sources are consulted and their scope is limited in

providing a context for the investigation, to assist with the
planning or execution of laboratory or field work, or in
informing the interpretation of results.
There is a partial rationale for the choice of investigation. The

biological background to the investigation is developed to
some extent.
Information gathered from some relevant sources has some

bearing on the context for the investigation, and assists, in a
limited way, with the planning or execution of laboratory or
field work, or to inform the interpretation of results.
The rationale for the investigation is clear, in terms of its

scope and relationship to biological principles.
Several relevant sources are consulted, and are used to

provide a context for the project, to assist with the planning
or execution of laboratory or field work, and to inform the
interpretation of results.
The rationale for the investigation is clearly justified in terms

of its scope and appropriate biological principles are
discussed.
Additional sources, beyond those that were most readily to

hand or were initially suggested by the teacher are selected.
The material chosen is selected for its relevance to the
investigation and it is used effectively to provide a context for
the project, to assist with the planning or execution of
laboratory or field work, and to inform the interpretation of
results.
Total marks
2 marks
4 marks
6 marks
8 marks

37
Exemplar 1 Jennys work

The effect of different concentrations of alcohol on
streptomycin
Abstract:
Within this experiment the relationship between different concentrations of alcohol and the
effectiveness of antibiotics is studied. This is done using lawns of bacteria, agar
impregnated with different alcohol concentrations and antibiotic discs. The results showed
an increase in the efficiency of the antibiotic with increase in alcohol concentration although
the result was not significant at the 95% level.
Introduction:
Does it matter if you drink alcohol whilst on antibiotics? Does it have any direct affect on
the antibiotic? Does the amount you drink have any correlation with antibiotic efficiency?
It is generally accepted that alcohol should not be consumed whilst taking any form of
antibiotic, but is this fact or an old wives tale?
This experiment uses the antibiotic streptomycin, the bacterium E.coli and absolute ethanol
at different concentrations to investigate these questions.
After preliminary research, it was recognised that high concentrations of alcohol would
interfere with bacterial growth and would never be found in the body. I therefore
researched the legal limit for alcohol concentration in the blood and decided to centre my
experiment on these values. The values are 50mg per 100cm3 in some countries and 80mg per
100cm3 in others (17).
Streptomycin (above) is a human antibiotic drug derived from the soil bacterium
Streptomyces griseus and is active against both Gram-positive and Gram-negative bacteria.
It was originally isolated by Selman A. Waksman and Albert Schatz in 1947 and was found to
be effective as a tuberculosis therapy (5). Streptomycin belongs to a group of drugs called
aminoglycosides, which are effective against aerobic bacteria, viruses and fungi. This
antibiotic works by inhibiting protein synthesis and damaging cell membranes in susceptible
microorganisms, although it is not clear exactly how (5). It passes into the cell through Porin
channels and attaches to the cells ribosomes causing it to produce ineffective proteins that
are vital for the cells survival, so the cell dies (10). This antibiotic seemed appropriate to
use in the experiment as it is effective against the bacterium E.Coli and is a human drug. It
is likely, therefore, that both alcohol and this antibiotic could be present in the blood of the
body.
The bacterium used was Eshcherichia coli. In optimal conditions this bacterium can divide
every 20 minutes, so can multiply from one cell to 108 in just 12 hours. It contains a 5 million
base pair chromosome, so the copying of DNA has to be fast and accurate to result in only
one mutation every billion nucleotides. Within the body E.Coli can be found living harmlessly
in the large intestines of humans, living off any unabsorbed organic material (16).
38

Preliminary Experiments.
Aims:
To ensure the alcohol does not kill bacteria at these concentrations.

To make sure the agar will solidify with the 5cm of solution.
Bibliography:
All websites accessed in March and April 2004

1. http://techniques.geog.ox.ac.uk/mod_2/tables//_pearsons_full.htm
2. http://health.iafrica.com/doconline/qa/pharmacology/800528.htm
3. http://www.acad.org.uk/04.html
4. http://pmep.cce.cornell.edu/profiles/exotoxnet/pyrethrins-ziram/streptomycinext.html
5. http://www.factmonster.com/ce6/sci/A0846951.html
6. http://www.atdn.org/access/drugs/stre.html
7. http://www.cancer.org/docroot/CDG/content/CDGstreptomycin sulfate.html
8. http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&listiuds=1037
5408&dopt=Abstract
9. http://www.wikipedia.com
10. http://www.chclibrary.org/micromed/00037050.html
11. http://www.madsci.org/posts/archives/1068575560.Bc.r.html
12. http://ntri.tamuk.edu/cell/ribosomes.html
13. http://www.sciencestuff.com/nav/instructions/agar3.htm
14. Salters-Nuffield student book 2
15. Introduction to advanced biology Clegg
16. Biology sixth edition - Campbell and Reece
17. The Highway code - The stationery office
18. Chemistry data book- Longman
19. Micro-organisms and biotechnology - Taylor
Total: 6/8
Moderators comments
Strand (a) the student has used biological principles to give a clear reason for
doing the work. Sources were referenced to help develop the rationale.
Strand (b) some sources were referenced to help plan the work and interpret
results that were not predicted.
Strand (b) was better than strand (a).
use research methods to help make some suggestions as to how alcohol might
inhibit the action of streptomycin
justify the use of E-coli as a model.

39
Exemplar 2 Michaels work

Investigating the antibacterial effect that an ivy extract has
on Bacillus subtilis
Abstract
This investigation was designed to demonstrate the antibacterial properties of ivy, this was
done using a bioassay technique by pipetting 0.4mls of B.subtilis onto an agar plate and then
carefully placing on the plate six pre-impregnated filter disks with different concentrations
of 0% (control), 25%, 50%, 100%, 200% and 330% ivy extract. This was repeated on five
other plates. The effect the ivy extract had on the bacteria could be seen after two days.
The results showed that the lower concentrations 25+50%, encouraged bacteria growth
where as the higher concentrations 100-330% did not stimulate bacteria growth but did not
kill it either showing a bacteriostatic effect. The higher the concentration after 100% the
smaller the bacteria growth around the filter disks. This method is easy to follow and
implement. The ivy extract showed an effect on the bacteria, although it was not
antibacterial at these concentrations, at the higher concentrations it showed a diminishing
bacteriostatic effect and if concentrations were increased I think a bactericidal effect may
well be found. If ivy extract was used as an antibacterial wipe the starting extract solution
would have to be reduced to less than half the original volume.
Aim
The aim of this investigation is to demonstrate the antibacterial properties of ivy leaves.
Rationale
The majority of medicine originates from plants, the World Health Organization estimates
that 75-80% of the worlds population uses plant medicines either in part or exclusively, an
example of this is aspirin which was derived from willow bark [1].
I have chosen the ivy plant as it is available all year and there is a sustainable source of it
nearby. I will be using the ivy leaves rather then the stems or roots as using the stems or
digging up roots could kill the whole plant.
I think a bioassay technique would be very useful if you were trying to find a selection of
plants that could be antibacterial if you needed to have some sort of wipe to contain an
infection and you were in an unknown area without medical support. This experiment is
therefore designed to give results over a short period of time that are decisive. In this
investigation I will be using a bioassay technique where by the presence of the ivy extract is
quantified by using living organisms. This will then show the strength of the antibacterial
effect of ivy by showing areas of inhibition around the filter disks.
The active ingredient must be bactericidal to produce zones of inhibition on nutrient agar.
This would show that the bacteria in a wound would be killed and stop an infection [11].
Ivy is a bitter aromatic herb; its leaves contain the compound emetine and triterpene
saponins and are antibacterial [4].
40

This bacterium is not considered pathogenic or toxic to animals, humans or plants therefore
suitable for this experiment. They are rod-shaped bacterium and have a flagella and are
gram positive in the early stages of growth [2]. This bacterium can be cultured on solid agar.
A culture B.subtilis will be grown on agar in a Petri dish on which discs of filter paper,
impregnated with different concentrations of ivy extract, will be placed. The key to getting
good results will be preparing and doing the experiment in as sterile environment as possible
in a school environment.
Null hypothesis: the ivy solution will have no effect on the bacteria this would be indicated
by having normal bacteria growing right up to the filter paper. There will be no significant
difference between the two means.
Working hypothesis: the ivy extract will have a negative effect on the bacteria growth
producing zones of inhibition as the ivy extract should contain a substance that is
bactericidal.
Bibliography
[1] Salters-Nuffield Advanced Biology book 2, (Heinemann 2002), pg134
[2] www.epa.gov
[3] Advanced Biology, Michael Kent, 2000, pg370
[4] www.scs.leeds.ac.uk
[5] Geography An Intergrated Approach, 2000, pg 636 for Spearmans rank significance
graph.
[6] Cleapss laboratory handbook, 1989
[7] Salters-Nuffield Advanced Biology book 3 (Heinemann 2003), pg122
Total: 7/8
Moderators comments
Strand (a) the rationale is clear and related to appropriate biological

principles. However, these could have been developed further.
Strand (b) additional sources were carefully selected and were used to help in
planning the work and in discussing the results.
develop the biological principles of bacterial agents and/or bacterial growth.

41
Assessment criteria Planning

B
Planning
There is some attempt to plan and to select the method or

apparatus chosen. Some relevant variables are identified.
Some potential safety hazards and the steps to avoid or minimise

them are identified.
A trial experiment may be carried out.
There is a plan for the investigation, with some explanation of the

selection of apparatus and methods. There are some details of how
variables are to be controlled, manipulated or taken into account
and how relevant observations are to be made.
Most potential safety hazards, and the steps to avoid or minimise

them, are identified.
A trial experiment is performed that has some bearing on the

planning of the project.
There is a clear plan of action, both for an initial trial phase and for
the main period of data collection. Apparatus selected and methods
chosen are appropriate to the investigation. There is discussion
about how variables are controlled, manipulated or taken into
account and about the collection of relevant observations or data.
All potential safety hazards are identified, and suitable steps taken
to avoid or minimise them.
A well-thought out trial is conducted in advance of the main data

collection phase, and is used to inform the planning of the
investigation.
a, b, c There is evidence of thought and ingenuity in the design of

experiments or the recording of data, with good attention to
detail including the way in which variables are controlled,
manipulated or taken into account and how relevant
observations are made or data collected. Apparatus is devised
or modified to suit the project as necessary.
42
Total marks
2 marks
4 marks
6 marks
8 marks

Exemplar 3 Samirs work

A study to investigate differing pHs on the growth of pea
plants
Aim:
To investigate the affects of pH on the growth of pea plants.
Introduction:
In this investigation I intend to study the affect of pH on pea plant growth therefore
ascertaining the best soil pH growing conditions. To carry out this experiment I must
understand the mechanisms in a plant that are affected by differing pHs. I chose to study
this investigation because of its significance in everyday life and pollution as an increasing
concern.
By studying the affect of pH on plant growth I am also therefore studying and acquiring a
small amount of knowledge into the affects of acid rain.
It is the industrial processes like the production of steel and iron and burning fossil fuels
that are the main contributors to acid rain. This is because they produce the gases sulphur
dioxide (SO2) and nitrogen dioxide (NO2) which are primary causes of acid rain. Acid rain
can be defined as a general term encompassing dry deposition and precipitation which has an
increased acidity due to contamination by carbon dioxide, sulphur dioxide and nitrogen
dioxide. (K.Byrne Environmental Science)
It is when nitrogen dioxide and sulphur dioxide react with the water and oxygen in our
atmosphere form an acidic solution, which falls back down to earth as acid rain.
Sulphur
SO2(g) + O2(g)
SO3(g)
Dioxide
SO3(g) + H2O (l)
H2SO4(l)
Nitrogen
3NO2(g) + H2O (l)

NO + NO2 + H2O
2H + 2NO3 +NO
Dioxide
2H + 2NO2
When deciding on a range of pHs to do in my experiment I took my inspiration from both

previous investigations and nature. I found values as low as pH3 have been recorded in
Southeastern USA (environmental protection agency). And pHs as high pH12 in the saline
lakes of Kenya/Tanzania (B.E.Jones, W.D.Grant).
I decided to do a range of pH 2 pH10 as a compromise.
To carry out this experiment it is essential to understand the mechanisms within a plant
that may be affected by any changes in pH.
Background information:
Photosynthesis:
6CO2+ 6H2O
C6H12O6 + 6O2
Plants gain their source of energy and material to synthesise their growth materials in form
of glucose, made by the light requiring process of photosynthesis.
There are two parts to the photosynthetic process; both take place in the choloroplasts
inside the cell. The two stages are the light dependent stage, which takes place in the
thylakoid membranes of the grana, and the light independent stage.

43
The Calvin cycle, in the light independent stage, relies on the help of enzymes where ATP
and reduced NADP help turn carbon dioxide into glucose, amino acids, lipids e.t.c which
provide the plants with its food.
The Calvin cycle: (diagram omitted)

The Calvin cycle cannot take place without the action of the enzyme ribulose biphosphate
carboxylase (RuBISCO), the commonest enzyme in the world. The RuBISCO fixes carbon by
combining Ribulose Biphosphate (RuBP) with CO2. Studies have shown that the optimum pH
for the enzyme RuBISCO is 8.2.
(www.clas.ufl.edu/users/gbowes/bot3503/psdkrx.html)
If the optimum pH is 8.2 then pHs significantly higher or lower will affect (denature) the
enzyme and therefore the rate of photosynthesis.
If the rate of photosynthesis slows down it will decrease the rate of production of glucose,
amino acids and lipids. This will decrease the growth rate of the plant.
Enzymes:
Enzymes can be described as biological catalysts as they catalyse (speed up without

changing) almost every metabolic reaction. A factor affecting the ability of enzymes to
fulfil their potential is pH. Different enzymes work best at different pHs, for example,
pepsin works best in the acidic environment of the stomach.
Each enzyme has an optimum pH and extreme changes can cause a change in the ionisation
state of the amino acid side chains in the enzyme molecules and therefore a change in the
ionic bonds within the tertiary structure of the enzyme (protein molecule) creating a change
in the shape of the molecule and its active sites, inhibiting the action of the enzymes on the
original substrate by rendering them unable to bind with the substrate molecules.
Enzymes are globular proteins, meaning they are protein molecules (chains of amino acids)
coiled into specific 3 dimensional shapes, creating their distinguishing feature, the active
site. This site is a particular shape to which the targeted substrate can bind, formed by the
bonding of various groups along the amino acid chain (ionic bonds, disulphide bonds, hydrogen
bonds and hydrophobic bonds). This is called the lock and key principle and is one theory of
how an enzyme works.
Another theory is the induced fit model. This puts forward that the walls of the active site
move a little to wrap around the substrate, this stretching putting stress on the active site
and the bonds within the substrate making them weaker and therefore lowering the
activation energy for the reaction. Apart from pH, enzymes are affected by a number of
other factors, including temperature because if too cold they do not have enough energy to
collide with their substrate or enough activation for it to be a successful collision, if too hot
then the enzymes denature as the hydrogen bonds within the active site break, changing the
shape of the molecule.
Respiration:
6CO2 + C6H12O6
6CO2 + 6H2O
Respiration relies on the use of enzymes to catalyse the metabolic reactions taking place.
Through the process of respiration plants gain their energy used for, among other things,
growth.
44

It is the Krebs cycle and oxidative phosphorylation which, massively rely on the action of
enzymes. During oxidative phosphorylation an electron carrier chain made of cyctochrome
(protein molecules) transports electrons down various energy levels to eventually be received
by an oxygen molecule. During the Krebs cycle, Acetyl co-enzyme A (ACoA) is combined with
a 4 carbon acid to form a 6 carbon acid which is converted (via a series of enzyme controlled
reactions, through various intermediates) to Oxaloacetate and eventually back to a four
carbon acid which combines with another ACoA to complete the cycle.
The protein molecules would be affected by a change in pH, altering their three dimensional
structure, leading to an interruption of the electron transport chain and therefore (due to a
feedback system) the whole of aerobic respiration.
Hypothesis:
The growth of pea plants will be affected by differing pHs (measured by the average length
of shoot (of three plants) and by the dry masses of each plant).
The greatest amount of growth will occur at pH7 and the least amount will occur at the two
extremes, pH2 and pH10.
Null hypothesis:
There will be no difference in the growth of the pea plants at different pHs and any
difference will be due to chance only.
Method:
Eight plant pots were filled with compost and three pea plants (each seed from the same
female parent plant) were placed an equal distance apart from each other and the sides
of each pot.
The eight plant pots were then labelled pH2, pH3, pH4, pH5, pH6, pH7, pH9 and pH10.
A mixture of 20cm3 of the corresponding acidic solution/distilled water was distributed
as evenly as possible (to make sure each plant is receiving the same amount of water also
using a measuring cylinder) over the three seeds in each pot,
The eight plant pots were then placed in the same greenhouse making sure they are all
receiving the same amount of light and are not shadowed by other objects.
Over the next four weeks all the plant pots were watered with distilled water at the same
time as each other (when they looked as though they needed water) with the same amount
to each (10cm3).
At various intervals during the weeks, the pea plants/shoots were measured and the
data recorded on a table.
Every seven days each plant pot was given 10cm3 of the corresponding acidic solution.
At the end of the four weeks the plants were carefully removed from the pots and the
soil was washed away with a gently running tap.
The wet weights of each plant was taken the plants were then placed in a pre-weighed
crucible and the crucibles were placed in a drying oven for 24 hours and then reweighed.
The values were then compared, length of shoot/plant and wet and dry masses at each pH.
Extraneous variables:
The genetic make up of each pea plant cannot be controlled so that they are each exactly
the same (unless each pea was a clone of one original pea). So difference in height of the pea
plants may be purely due to the fact that one plant had the genetic makeup coding for it to
be short and therefore nothing to do with the varying pHs.

45
Controls:
The temperature and amount of light exposure was the same for each plant pot as they
were all kept in the same greenhouse on an open shelf where they didnt experience and
shadowing from other plants.
The amount of water given to each plant pot was the same (10cm3).
The volume of acidic solution given to each plant pot was the same (10cm3 each week).
The amount of nutrients the plants received was kept the same by using the same
compost (vermiculite) and the same amount was used in each plant pot.
All the peas came from the same parent plants to make them as genetically similar as
possible.
Three pea plants were grown at each different pH so as to allow for varying effects of
the different genetic make-up of the plant.
Each plant was watered and measured at the same time.
The distance apart from each other and the sides of the plant pot was the same for
each plant (as far as possible) so that each pea plant had the same amount of space in
which to grow and were also therefore competing for the same amount of as each other.
Each pea plant pot was grown for the same amount of time i.e. four weeks.
Safety precautions:
As the experiment involved dealing with an acid, care was taken when handling the acid
and eye protection was worn.
Total: 4/8
Moderators comments
Strand (a) the student had a clear plan of action but some details were not
considered or carried out. For example, the layout of the plants in the
greenhouse should have been randomised, vermiculite is described as a compost
and volume should have been used instead of amount.
Strand (b) one relevant safety precaution is stated. This experiment does not
have major hazards but a little more might have been added.
Strand (c) there is no record of a trial experiment.
report the trial experiment
increase the number of pea plants given each treatment
obtain more performance data from the pea plants, such as leaf length or leaf
area, as evidence of good attention to detail.
46

Exemplar 4 Robyns work

An Investigation into the Anti-bacterial Properties of Allicin
supplements.
Abstract:
This experiment was done in order to investigate the antibacterial properties of garlics
active ingredient, allicin. It was also to see if allicin could work in synergism with the
antibiotic streptomycin. It was found that at low and high concentrations allicin inhibits the
antibiotic and then at high concentrations is starts to have its own anti-bacterial effect.
Research and Rationale:
It has long been believed that garlic is good for colds and general sickness; I decided to
make the aim of my investigation to determine if allicin, garlics active ingredient, has any
anti-bacterial properties. I found a supplement that is 100% allicin and its website [see
bibliography] claims ALLIMAX may be used to reduce symptoms of bacterial infections and
also kills E. coli. A garlic supplement is often recommended by herbalists as a supplement
worth taking whilst on antibiotics, so this investigation is seeing if there is any truth in this.
Garlic [Allium sativum]; produces allicin when its injured through an enzyme based reaction
possibly to protect itself from insects and fungi.
Allicin is known as 2-propene-1-sulfinothoic acid S-2-propenyl ester; thio-2-proene-1-sulfinic
acid S-allyl ester. The enzyme, alliinase, is stored in separate compartments to the
compound alliin in garlic. When its damaged the two combine and produce allicin:
[picture from: www.allicin.com]

Allicin is theorised to destabilise enzymes that are used by bacteria to infect the body by
attacking their active sites particularly the sulphydral groups by forming disulphide bridges.
[See bibliography from information source]
Bacterial cell walls contain a unique material called peptidoglycan, this is made of polymers
of modified sugars cross-linked by short polypeptides that vary with species, a build up of
peptidoglycan results in a single laminated structure that encloses and protects the
bacterial cell from attack. The aminoglycoside antibiotic used in this investigation is
Streptomycin which works by entering through the bacterial cell through an oligopeptide
transport system in its membrane. Once inside the cell it prevents the two units of a
ribosome from joining together, and therefore preventing protein synthesis.

47
[picture from website (see bibliography)]

The aminogycoside, in this case streptomycin, prevents the larger part of the ribosome from
joining onto the smaller and therefore engaging in protein synthesis. [see bibliography for
information source]
Allicin is highly unstable and quickly changes into other products. ALLIMAX is a one-a-day
supplement that contains 180mg of allicin in each capsule.
As allicin is meant to be a supplement I decided to test it with an antibiotic, to see if allicin
aided antibiotics works better than antibiotics alone.
The best agar after testing with 3 drops of E.coli spread on each with aseptic technique
[see appendix note i] I found to be Nutrient Agar.
Planning
Preliminary:
My first plan was to dissolve the allicin capsules in water with pepsin to break down the
actual capsules. 2 molar hydrochloric acid was added to simulate the acidic stomach
conditions that the Allimax tablet first comes into and to give the pepsin its best working
conditions. There were different amounts of water to make the capsules dilute to different
concentrations.
Individual agar plates that had been prepared by aseptic technique [see appendix note i].
Another aspect of the investigation was to determine if the allicin would work better with
the antibiotic inside the agar or on top of it. So I made two of all the following:
Quantity of water
10 cm
15 cm
20 cm
25 cm
30 cm
Quantity of pepsin
1 cm
1 cm
1 cm
1 cm
1 cm
Allimax tablets
1
1
1
1
1
The pepsin solution = I spatula of pepsin and 20cm of HCL

Whilst pouring the agar plates in my aseptic technique [see appendix note i] I added one set
of the final mixtures in the table to the agar and then let them set, the second set of the
above mixtures I spread over the top of the agar plates after they had set. I then spread 3
drops of E.coli over every plate and placed them into the incubator over night at no less than
35oc. I then measure the growth of E.coli by tracing the growth pattern onto a piece of OHP
transparent sheet and then put graph paper underneath and counted the millimetre squares
so that I end with an inhibition area in mm2.
48

The results showed that mixing the allicin into the liquid agar somehow inactivated it or
didnt allow it to reach the E.coli. With the other dishes with the allicin spread on top the
concentrations of the allicin seemed to be too weak as the inhibition was only just
recordable. The other problem with this was that the gelatine capsule didnt completely
break down and the solutions therefore had to be filtered, this was difficult and time
consuming as it had to be done with each test tube with new filter paper.
As a result of the preliminary test [see appendix note ii for results] I decided that I should
investigate the concentrations that would be found in the blood and work from that point to
find appropriate concentrations. [See observing and recording for calculations of
concentrations] I also decide that the filtering of the solutions was having an adverse
effect so I decided that I should split the Allimax tablets in a sterilised plate and rinse the
powder out. I basically evolved the following method:
Method:
One Allimax tablet is cut into using a sterile [see appendix note i] scalpel and emptied out
onto the top of an empty agar plate.
It then has to be washed down into the 1 litre volumetric flask with distilled water.
Then make up the mark in the volumetric flask with the distilled water, but stop to shake
the volumetric flask periodically so that the allicin mixes in with the water.
Then 5 beakers have the following placed into them:
Beaker:
1
2
3
4
5
Quantity of allicin:
10 cm from the 1L conc.
Quantity of water:
10 cm
15 cm
20 cm
25 cm
30 cm
Then place 3 antibiotic disks into each of the beakers to soak.

Then using aseptic technique [see appendix note i] prepare six agar plates with nutrient agar
and let them set.
Next put three 0.5cm2 drops of E.coli onto each plate.
Then place 3 antibiotic streptomycin disks onto one of the E.coli agar plates to act as a
control, then with a pair of sterile forceps take the antibiotic streptomycin disks from
beaker one and place then onto a different agar plate with the E.coli.
Repeat this step with beakers 2-5 and the other E.coli agar plates.
The plates should then be incubated overnight at no less than 30oC and then placed in the
refrigerator in the morning to stop growth.
The results were recorded by:
First tracing the areas of inhibited growth and the ring onto a piece of OHP transparent
sheet.
Then place a piece of graph paper underneath and count the squares of the inhibition area.
Total: 7/8

49
Moderators comments
Strand (a) this was marred by the use of a spatula of pepsin rather than a
defined mass. Further details of some aspects of the methods were given in an
appendix, eg aseptic technique.
Strand (b) safety hazards are identified in the appendix. However, these should
have been given a little more emphasis in the report.
Strand (c) the student gave thought to the trial experiments and there is
evidence of attention to experimental details.
describe the control of variables as a section of planning
make some comments on safety in the plan.
50

Assessment criteria Implementing

C
Implementing
Apparatus is used with some skill and attention to health and

safety.
Some previously learned techniques and procedures are carried out

appropriately.
The initial plan has some bearing on the execution of the work.
Apparatus is generally used appropriately and with confidence, care

and skill.
Techniques are usually applied correctly and extended or modified

if necessary. Work is generally methodical and well-organised.
There is some reviewing of the initial plan as the project proceeds.
Apparatus is used confidently, carefully and skilfully. There is good

awareness of health and safety and all reasonable steps taken to
minimise risk throughout the investigation.
Techniques are always applied correctly and extended or modified

if necessary. Work is always methodical and well-organised.
The initial plan is reviewed frequently in the course of the project,

and modified or maintained according to results obtained.
Total marks

2 marks
4 marks
6 marks
51
Exemplar 5 Marcos work

A Comparison of Insect Species Diversity of an Uncut Meadow
with a Woodland Area
Abstract:
In any woodland where succession has taken place, an ecosystem exists. Within the
ecosystem, only certain species can survive due to the role of survival of the fittest.
In a meadow which is left to grow, a different ecosystem exists, maybe with different
species able to survive in it.
This study is aimed at investigating whether the same species can exist in two different
ecosystems and whether the species diversity of those ecosystems are different. This will
be done using the method of pitfall trapping. These traps will be used to collect and count
the number and type of species in each ecosystem at 15 places in each site.
Tests were carried out on the data to compare the two groups. A Simpsons diversity Index
was found for the two sets of data. It was found the species diversity of the meadow was
larger than that of the woodland. A Mann-Whitney U-Test was also carried out on the sets
of data. This showed that there was no significant difference in the two sets of data,
proving the null hypothesis.
Hypothesis:
I hypothesis that there will be a higher species diversity in the uncut meadow that in the
woodland. My belief comes from the fact that in a woodland, the light intensity in the
woodland will be less than in the uncut meadow.
Null Hypothesis:
My null hypothesis is that there will be no significant difference between the two sets of
data from the two sites.
Method:
Find suitable sites to set the pitfall traps, possibly away from obstructions that may
aggravate the grid system that I will use.
Set some pitfall traps near the area that I have chosen. This will help to perfect the
technique of planting them and possibly catch a few samples of insect, to gain an idea of
what I should expect to catch.
Calculate grid numbers using the purpose-made computer program.
To keep the results accurate and reliable, continuity must be using in planting the traps and
uprooting them the next day. The following method was followed to ensure that this
happened.
52
Place a marker where you plan to place the origin of the grid.
Measure 90 to the right for the x-coordinate.
From the x-coordinate, measure until you reach the specified y-coordinate.
Set the pitfall trap in the ground at that point, being careful to set it following the
method.
Set all of the pitfall traps in that area and note the time that you finished.
Use the same method to set pitfall traps at the next site, being careful to note the time
that all the traps were set.
During the next day, start checking the pitfall traps at the time that you finished
planting the traps for that site. This will ensure that all of the traps have been set for
24 hours.
Check the next sites traps at the time they were finished being laid.
At each cup, the contents should be emptied into a deep tray. Using a magnifying glass,
identify each species that was caught in the trap. Once this has been done, empty the
contents of the tray in the same area as your traps, but away from them so the same insect
is not caught twice. In the event that the species is unidentifiable with the reference that
you have, take the insect, in a sample pot, back to the field centre and use extra references
to identify the insect. When this is done, place the insect back into the test site.
Ethics:
The main ethical question that may arise from this investigation is that catching the insects
and only checking the traps the next morning may leave them without a food source for up
to 24 hours. Some insects may be able to survive this but, inevitably there may be some
death of insects. This is unfortunately unavoidable and so if an insect dies, it should be
placed back into its original environment to be either eaten by one of its prey or digested by
soil-dwelling bacteria.
The hole that is made for the pitfall trap must also be filled in after the trap is emptied.
This prevents any insects or mammals from falling down them once the investigation is
finished.
Variables:
The main variable that I am testing is the effect of sunlight on the species diversity of 2
areas. The sunlight will affect the species diversity because if there is more sunlight, more
biomass will exist in the area, meaning more species can survive there. With less sunlight,
more specific insects will live where only some plants can live.
Pitfall Traps:
To carry out this investigation, I will use pitfall traps, set in a grid of 5m x 5m, to catch the
insects that inhabit the ecosystems of each site, the woodland and the uncut meadow. Pitfall
traps are one of the most effective methods of capturing ground-living insects
(www.marion.ohio-state.edu). A smooth-sided container is buried flush with the ground. The
insects will walk into the trap and will not be able to climb out because of the smooth sides.
The best and cheapest way of pitfall trapping is to use disposable plastic cups. The sides are
smooth enough so the insects cannot climb out and they are cheap and disposable.
To lay a pitfall trap
Dig a hole in the soil using the soil corer
Collect the soil in a bucket for use later
Place a cup in the hole. The cup should be flush with the ground and any gaps should be
filled in with spare soil from the bucket
Once the trap has been used, remove the cup and fill in the hole with soil from the
bucket
Apparatus:
The apparatus I will use are:
30 plastic cups (used as pitfall traps)

53
5m measuring tape (used to mark the x-axis of the grid)

metre rule (used to measure up the y-axis)
soil corer (used to make the hole for the pitfall trap)
Trial Experiment:
Before carrying out this investigation, I lay 10 pitfall traps in random places in the meadow,
well away from the site I had chosen to carry out my actual investigation. This trial
experiment was to familiarise myself with the working of the apparatus and to practice
laying a pitfall trap.
Total: 4/6
Moderators comments
Strand (a) the apparatus was simple, but seems to have been used with
confidence.
Strand (b) the work seems to have been carried out methodically.
Strand (c) there is no evidence of the initial plan having been reviewed.
describe how to minimise the health risk to students
present a clear map to show the location of the samples
repeat the trapping or leave the traps for longer to catch greater numbers of
organisms as evidence of reviewing the plan.
54

Exemplar 6 Shazias work

Investigating the effect of flow rate on the diversity and
abundance of organisms in a freshwater river
Rationale:
The freshwater world has more than 500,000 different species of insects. They occur in
habitats that range from hot springs, discarded tin cans, temporary ponds, rivers, lakes to
arctic and mountain pools. Take a walk down to a lake. It doesnt matter what time it is
there will be insect activity somewhere!
R Narf, 1997 (1)
Freshwater streams contain a wealth of organisms many onlookers take for granted.
Freshwater streams have a salinity of less than 0.005% and contain a number of distinct
microhabitats. Successful freshwater organisms must be able to withstand the hazards and
exploit the benefits of their habitat to survive. The organisms present depend upon three
main factors, depth, flow rate and chemical composition of the water. (2)
Stream water travels at a particular velocity and this has a great bearing on the organisms
that occupy the ecosystem and the ecological niche in which they exist. (6) Organisms living
in fast flowing streams will have evolutionary adaptations acquired through natural selection
that enable them to survive. Some of the adaptations are listed below:
1) Suckers used as a temporary means of attachment.
2) Hooks/claws used to cling onto rocks.
3) Streamlining to reduce drag.
4) Body-flattening.
5) Flight.
Null Hypothesis:
Biodiversity within the river will be constant regardless of flow rate. The abundance of
freshwater organisms within certain areas of the river (the areas being equal in size) will be
constant regardless of flow rate.
Preliminary Work:
My preliminary work will involve choosing two sites at which to make a comparison. It is
important that as many variables as possible, preferably all the variables, remain constant to
maintain a fair test, enabling a fair comparison to be made. I have decided to compare two
parts of Totley Brook River which have different flow rates, therefore many variables are
likely to be constant e.g. the chemical composition of the water.
After visiting the river I decided the areas being investigated should be approximately 5
metres in width, as this was the average width of the river, and approximately 5 metres in
length. This will give a large enough area to obtain a fair representation of the river.
Site
Width (m)
1
4.95
2
5.00
3
4.85
4
5.10
5
5.05
Average (m)
4.99
I determined the flow rate of the 5 sites as follows:

1. Using a tape measure mark out a length of 5 metres.
2. Place a cork in the river at one of the marked ends, start a stop clock immediately.
3. Stop the stop clock when the cork has traveled 5 metres.
4. Repeat 4 times, find the average. This ensures the result obtained is accurate and
reduces the amount of error in the final value.

55
Site
Time (s)
Average Time
(s)
Speed
(m/s )
1
43.89
43.58
43.43
43.98
43.54
43.7
2
60.80
60.47
60.39
60.57
60.40
60.5
3
20.70
20.59
20.65
20.31
20.46
20.5
4
20.59
20.40
20.49
20.96
20.60
20.6
5
22.70
22.39
22.67
22.59
22.19
22.5
5 / 43.7 =
0.114
5 / 60.5 =
0.0826
5 / 20.5 =
0.244
5 / 20.6 =
0.243
5 / 22.5 =
0.222
The average time and speed are given to 3 s.f.

After determining the flow rates I decided to compare sites 2 and 4 as the flow rate of site
2 is approximately 3 times slower than site 4. This is also true for site 3, however the
difference in widths of sites 2 and 4 is smaller, only 0.1m. (The difference in width of sites
2 and 3 is 0.15m). By comparing sites 2 and 4, a fairer comparison can be made, leading to
more reliable results.
From now on site 2 will be referred to as site 1 and site 4 will be referred to as site 2.
Data was then collected on the remaining variables. Light intensity was measured using a
light meter. Dissolved oxygen, p.H. and temperature were measured using a multimeter that
had a corresponding probe for each variable. The depth was measured using a metre ruler.
Measurements were taken at 1 metre intervals. As the variables can vary, it is important
they are as close to constant as possible so a fair comparison can be made and for the
experiment to be a fair test. If the variables differed, a difference in abundance or
diversity would not be due solely to flow rate, but also due to the differing variable e.g. if
site 2 was faster flowing and more alkaline than site 1 but the other variables remained
constant, then it would be a combination of the 2 variables that caused the difference in
abundance or diversity.
Site
Light intensity (lx)
Dissolved oxygen (mg/l)
Temperature ( C)
p.H.
Depth (m)
0
1
2
3
4
5
1
4125
9.6
10.6
9.2
2
4300
11.3
10.6
9.3
0.07
0.09
0.10
0.29
0.35
0.08
0.04
0.07
0.16
0.25
0.20
0.11
The amount of dissolved oxygen differs at each site, which is expected. The oxygen content
and flow rate at site 2 is greater than site 1. Any oxygen used by freshwater organisms is
replaced due to a constant influx of water rich in oxygen resulting in an increased oxygen
content. The water at site 2 is not replaced as frequently therefore any oxygen used by
freshwater organisms is not being replaced as often resulting in a reduced oxygen content.
56

The amount of vegetation also affects oxygen content, however I didnt have the amenities
or a large enough time scale to accurately determine the amount of plant cover/biomass and
its affect on oxygen content without destroying the site (and ecosystem).
In order to familiarize myself with the freshwater organisms at the 2 sites I carried out
one random kick sample at each site.
Hazards and Safety:

Ensure you wash your hands after carrying out the practical as the water may be
contaminated. Dont drink the water.
The Consideration of Living Organisms and the Ecosystem:

It is important to treat living organisms with respect throughout the investigation. As few
organisms as possible should be removed from their natural habitats. They should be
returned unharmed to the exact position they were taken from, as they will have adapted to
their habitat and may suffer/die if returned to an incorrect habitat. It is important to keep
the organisms and water cool as overheating, even by a few degrees, can rapidly kill
organisms. Keep collecting trays in the shade, away from direct sunlight and warmth.
Protect the ecosystem by not removing or damaging vegetation. If while carrying out a kick
sample stones are overturned return them to their original position. Dont drop litter.
Ensure you leave the site how you found it.
Practical:
Apparatus.
0.5m x 0.5m quadrat
Collection tray
Net
Ice-cube tray
Petri dishes
Pipettes
Plastic tea-spoons
Diagram:
Collecting tray
Quadrat
Ice-cube tray
Plastic teaspoon
Petri-dish

57
Method:
1) Randomly throw a 0.5m x 0.5m quadrat.
2) Hold the net in the stream and carry out a kick sample (i.e. kick the area within the
quadrant 20 times).
3) Empty the contents of the net into a collection tray by turning the net inside out. Wash
the net thoroughly with water from the stream to ensure that the net is completely
emptied.
4) Carefully sort through the rocks and debris in the collection tray and gently wash any
animals clinging onto rocks/stones into the tray.
5) Allow the water in the collection tray to settle. Using pipettes and plastic teaspoons
catch and separate the organisms into groups in separate compartments of an ice-cube
tray.
6) Transfer individual organisms into petri dishes and examine them. Record the number of
species and abundance of each.
9) Return all organisms to the stream.
10) Repeat 5 times ensuring the quadrat is thrown randomly within the site.
11) Find the average.
Results:
Num be r of
orga nism s in
qua dra t
S ite 1
S pe cie s
A
B
C
D
E
A verage
0
0
0
0
6
1
0
0
0
0
7
2
0
0
0
0
6
3
0
0
0
0
5
4
0
0
0
0
7
5
0
0
0
0
8
6
2
3
2
4
6
30
7
4
2
1
5
4
29
7
2
3
2
3
5
27
6
3
2
1
6
7
32
8
3
3
3
4
5
34
7
3
3
2
4
5
30
S ite 2
S pe cie s
A
B
C
D
E
F
G
H
I
J
4
1
1
1
0
0
0
0
0
0
5
2
2
1
0
0
0
0
0
0
6
2
2
3
0
0
0
0
0
0
6
0
3
2
0
0
0
0
0
0
4
2
1
0
0
0
0
0
0
0
5
1
2
1
0
0
0
0
0
0
Tota l
10
13
11
10
F
G
H
I
J
K
Tota l
Values in the table are to the nearest whole number.
Total: 5/6
58

Moderators comments
Strand (a) the apparatus is generally used appropriately.
Strand (b) the work seems to be methodical and well-organised.
Strand (c) the initial plan seems to have been carried out rather than reviewed
but the student did carry out a trial sampling to practise organism identification.
give more details about minimising risk
review the plan more clearly.

59
Assessment criteria Observing and recording

D Observing and recording
a
Some appropriate measurements and observations are recorded

which are adequate for the method used and reasonably accurate.
There is some repeating or checking of values obtained.
Measurements and observations are recorded methodically and

accurately in appropriate units, and some thought is given to
precision and repeatability.
A reasonable number and range of observations and measurements

are carried out. Any anomalous results are noted. There is some
appropriate modification of procedures for data collection if
necessary.
Observations and measurements are carried out over a suitable

range of values or conditions. Sufficient observations and
measurements are made to allow a conclusion. Numerical results
are recorded to an appropriate degree of precision.
Measurements and observations are repeated as appropriate. Any

anomalous results are noted and investigated. If problems arise in
the making of measurements or observations, procedures are
adapted to ensure data are reliable.
60
Total marks
2 marks
4 marks
6 marks

Exemplar 7 Susans work

Individual Investigation: Comparing the population of damselfly
nymphs in a managed and unmanaged pond.
Abstract:
This investigation was designed to compare the population of damselfly nymphs in a managed
and unmanaged pond. It involves keeping certain variables constant if possible. The abiotic
factors, light intensity and pH and oxygen availability, were measured and then the results
used as to whether any differences could account for the differences in the population of
damselfly nymphs in the two ponds. The results showed that in the managed pond there were
more damselfly nymphs than in the unmanaged pond. In the managed pond the light intensity
was a lot higher which could be the reason for the larger number of damselfly nymphs as the
light would benefit the vegetation and therefore increase the oxygen availability in the
pond, making it a better environment for the nymphs.
A t-test was used which proved to be significant because this value was higher than the
value for the degree of freedom, revealing that the results are 99.99% accurate.
Introduction:
This investigation took place at Scadbury Park. It is a LNR (Local Nature Reserve) covering
around 300 acres of land. It consists of meadowland, woodland and freshwater. Scadbury
Park has been named as a SSSI (Site of Special Scientific Interest). On visiting the park on
the first day, there was a recognisable difference between the light intensities of an
unmanaged pond and a managed pond so an investigation into the effect the light intensity on
the diversity of a certain species seemed interesting. The species studied was damselfly
nymphs because they are easy to identify and certain factors are likely to affect their
population.
Damselfly nymphs can measure from 2 to 3 cm in length and they live and breathe
underwater. They live among the weeds in still water and are carnivorous. (2) They catch
animals that come near them by the quick action of their jaws. They see animals coming with
their large compound eye that is made up of many single eyes. (1) They have wing buds, leaflike tracheal gills for breathing. (4) These are air tubes which collect oxygen from the
water. They have 6 legs and have 3 feathery tails that are flat or fan-like and usually oval in
shape. (5)
Planning:
The two ponds that will be investigated are a managed pond and an unmanaged pond. The
managed pond was a small manmade pond with a lot of vegetation squashed into a small space.
It covered most of the pond. This pond was in a very open space with no shelter.

61
The unmanaged pond was a lot bigger than the managed pond and duckweed covered the
surface. The banks were muddy and there were some reeds around the edge of the pond.
This pond was in the closing of the forest and so had shelter from the sun unlike the
managed pond.
A trial investigation was carried out to test out the necessary method. The results were as
follows:
Sample
1
2
3
No. of damselfly nymphs in

unmanaged pond
1
1
2
No. of damselfly nymphs in

managed pond
5
2
4
This showed that this investigation may show a significant difference and so the variables to
be kept constant and the variables to be changed were planned.
Without having done the pilot experiment, the method wouldnt have been adapted in order
to remove the duckweed without disturbing the wildlife underneath. It also seemed clear
that more samples needed to be taken to make sure that the difference seen in the trial
experiment was not anomalous.
A factor that could affect the data is the size of the pond and the depth of the pond.
Since the unmanaged pond is a lot larger, this may reflect in the results.
The independent variables are oxygen concentration, a chemical factor, pH, an edaphic
factor, and light intensity, a climatic factor. These variables are abiotic factors. (6) The
dependent variable was the number of damselfly nymphs and the controlled variables
were those such as using the same net, keeping the length of the sweep constant.
In order to analyse my data, I will use the t-test. This is the most appropriate test
because I will have a lot of samples to analyse and the t-test allows for that. From the
result of the t-test, I can then work out the reliability of my results and whether there
is a significant difference or not.
Hypothesis:
After the experiment had been planned, the null hypothesis was not likely to apply. The null
hypothesis says that there will be no difference between the populations of damselfly
nymphs in both ponds. The experimental hypothesis would apply in this case where the aim is
to prove that the managed pond has a higher population than the unmanaged pond. This is
because the managed pond has more plants and has more light reaching the pond therefore
meaning that more oxygen is in the water which would provide the damselfly nymphs with a
better environment to live in since they need oxygen to survive. Even though the unmanaged
pond has duckweed, there are not many plants that root from the bottom of that pond. Also,
the unmanaged pond does not get much light as it is shaded by the canopy.
Results:
This table contains the results from the experiment. 18 samples were taken from each of
the two ponds and the number of damselfly nymphs was recorded from each sample. From
this table a graph can be plotted and used to deduce which of the two ponds had the larger
population of damselfly nymphs.
62

Sample
No. of damselfly nymphs in the

unmanaged pond
No. of damselfly nymphs in the

managed pond
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
1
1
1
1
2
1
1
1
1
2
1
1
1
3
1
1
2
1
2
5
1
0
8
4
0
2
8
5
5
2
7
1
2
6
2
4
Figure 1: The number of damselfly nymphs recorded from each sample.

This table shows the results of the dissolved oxygen kit, the pH meter and the light
intensity meter.
Dissolved oxygen level (mg/l)
pH
Light intensity (LUX)
Managed Pond
6.9
7.3
17730
Unmanaged pond
6.8
7.3
8880
Figure 2: The oxygen level, pH and light intensity of each pond.

This frequency table shows the number of samples that contained a specific number of
damselfly nymphs.
No. of damselfly nymphs
0
1
2
3
4
5
6
7
8
No. of samples in unmanaged

pond
0
14
3
1
0
0
0
0
0
No. of samples in managed

pond
2
2
5
0
2
3
1
1
2
Figure 3: Frequency table

63
Frequency graph to show the number of samples that contain a

certain number of damselfly nymphs.
16
14
Number of samples
12
10
Samples in Unmanaged Pond
Samples in Managed Pond
6
4
2
0
1
Num ber of dam s elfly nym phs
Figure 4: The number of samples that contain a specific number of damselfly nymphs.
Total: 3/6
Moderators comments
Strand (a) the observations are recorded methodically and counting whole
organisms is an appropriate level of precision for this type of work.
Strand (b) the number of observations is only just adequate. Repeating the
whole experiment on another day was not discussed. The possibility of anomalous
results is also not discussed.
sample more ponds or sample the same pond over time
discuss the reliability of the data due to adapting the collecting procedure in one
pond.
64

Exemplar 8 Gabriels work

Investigating the effect of ethene on the ripening of fruit
Variables that need to be considered
Both containers need to be stored in the same place under the same conditions to ensure
the experiment is a fair test. The area where the containers are placed needs to be
cool and shaded as this will prevent other factors, such as sunshine and heat from
affecting the ripening of the fruit.
The size of the containers needs to be the same to ensure there is an equal volume of
space for the gases to diffuse into.
The weight of the bananas used need to be roughly equal and from the same bunch to
ensure that they are at the same stage of ripening.
The amount of strawberry tested needs to be weighed and be exactly the same for all
strawberries electronic scales can be used to ensure a weight of 15g is used in the
tests.
Use strawberries from the same pack to ensure similar levels of ripeness. However
these strawberries are still likely to vary in age and this should therefore be
considered when analysing results.
The experiments need to be carried out at the same time each day so that the period of
ripening between each set of experiments is the same.
Results:
The raw experimental results can be found in Appendix 6. Below are the average results that
have been produced as a result of the repetitions of each stage of the experiment.

65
Hours
0 hrs
24 hrs
48 hrs
72 hrs
96 hrs
Container
No ethene
Ethene
No ethene
Ethene
No ethene
Ethene
No ethene
Ethene
Benedicts test observations

Dark orange sugar present
Average Colorimeter reading

0.684
0.728
1.022
1.470
1.722
1.286
0.782
1.090
0.702
Anomalous results:
Some slightly anomalous results were produced in this experiment such as the colorimeter
reading of 1.38 on the strawberries without ethene after 24 hours, and also the reading of
0.43 on the strawberries after 96 hours. It is almost certain that the strawberries will not
all be at the same level of ripeness and this could have caused some of the anomalous
recordings. The riper strawberries will obviously have a higher sugar level than the others
and this explains the high reading of 1.38. Later on in the experiment there was a problem
with mould growing on and into the fruit. As the mould grew into the strawberries it had the
effect of changing the colour of the fruit to brown, in turn this had the effect of lowering
the colorimeter reading. This would therefore explain why later on in the experiment all of
the colorimeter readings have lowered, as reflected in the line graph, due to the growth of
mould on and into the remaining strawberries.
Appendix 5: Raw Data produced from the experiment

Results Day 1
Benedicts test - observations
Unripe strawberries beginning of
experiment
After 0 hours
1
2
3
4
5
Average
Colorimeter
Reading (OD)
0.73
0.70
0.65
0.59
0.75
0.684
Day 2
Strawberries without Ethene present
After 24 hours
1
2
3
4
5
Average
Strawberries with Ethene

After 24 hours
1
2
3
4
5
66
Colorimeter
Reading (OD)
0.87
0.77
0.71
1.38
0.91
0.728
Colorimeter
Reading (OD)
1.04
0.92
0.84
0.95
1.36
1 022

Day 3
After 48 hours
1
2
3
4
5
Average
Colorimeter
Reading (OD)
1.29
1.44
1.54
1.58
1.50
1.470
Total: 6/6
Moderators comments
Strand (a) sufficient observations were made to allow a conclusion and the
results were recorded with an appropriate degree of precision.
Strand (b) measurements were repeated and anomalous results noted and
investigated.

67
Assessment criteria Interpreting and evaluating

E
Interpreting and evaluating
There is some data processing. Statistical analysis is only completed

with detailed guidance. Application of calculated statistical values
is present, though limited or confused.
There is an attempt to apply biological principles.
Some conclusions are stated. There is some awareness of the

limitations of experimental results and conclusions.
Data are processed with some thought as to choice of method. The

chosen statistical test(s) may be inappropriate or provide limited
analysis of the stated hypothesis. Calculations are clearly set out
but the interpretation of calculated values lacks detailed
explanation. Some trends and patterns are identified.
Some attempt is made to interpret results using biological

principles, and to draw conclusions based on experimental results.
Conclusions are supported by the results. The limitations of the

results, and conclusions based upon them, are recognised. Any
limitations of the methods are recognised.
Data are processed using appropriate methods that reveal trends

and patterns. The chosen statistical tests are appropriate to the
data to be analysed and the hypothesis to be tested. Calculations of
statistical tests are clearly set out and interpreted, using a null
hypothesis and 5% confidence levels where appropriate. Trends and
patterns are identified.
Results are interpreted using biological principles and concepts of

Advanced GCE standard. Relevant biological principles are applied
correctly throughout.
Conclusions are supported by the results. The limitations of the

results, and conclusions based upon them, are recognised and
evaluated. Any limitations of the procedure are commented upon,
and sensible modifications suggested.
68
Total marks
2 marks
4 marks
6 marks

Exemplar 9 Jatinders work

The distribution of Glasswort on a salt marsh in relation to
the compaction of soil
Abstract:
The aim of this investigation was to determine the relationship between the growth of the
salt marsh plant Glasswort and the compaction of soil. The alternative hypothesis was that
there would be more glasswort growing in areas of high soil compaction, as only in these
areas can the plant become securely rooted against the tide. Statistical analysis showed the
results to be significant, proving that the alternative hypothesis was correct.
Alternative hypothesis:
More Glasswort will grow in areas of greater soil compaction.
Null hypothesis:
There is no link between soil compaction and Glasswort growth.
Prediction:
I predict that in areas of high soil compaction, more Glasswort will grow. This is because
Glasswort seeds need to be able to set out roots and become secure against the tide during
a period of only three days. As soil compaction levels decrease, less glasswort will grow, as
seedlings will find it increasingly difficult to take root.
Graph to show the relationship between the growth of Glasswort and soil compaction
100
90
frequency of Glaasswort
80
70
60
frequency of Glasswort
50
Log. (frequency of Glasswort)
40
30
20
10
0
0
10
11
12
cm into soil

69
Analysis:
My results show that Glasswort frequency is highest with high soil compaction levels. At the
central soil compaction levels there is variation in Glasswort frequency - it grows but with
only weak correlation between the frequency and the soil compaction. The results also show
that below a certain soil compaction level, no Glasswort grows at all.
Statistics test:
In order to be certain that my results are of importance, I have to know that they are over
95% significant.
To work out their significance I can use the Spearmans rank statistics test.
Spearmans rank:
Sample
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
sample
1
2
3
4
5
6
7
8
9
10
11
12
13
14
15
16
17
18
19
20
70
Distance into
soil/cm
6.9
9.9
6.1
4.2
4.7
4.4
3.2
1.6
9.2
1.2
4.0
5.7
5.8
5.9
4.5
6.6
5.6
4.2
8.2
3.3
rank 1
17
20
15
6.5
10
8
3
2
19
1
5
12
13
14
9
16
11
6.5
18
4
rank 1
17
20
15
6.5
10
8
3
2
19
1
5
12
13
14
9
16
11
6.5
18
4
rank 2
15
2
5
11
13
16
7
8
2
20
9
17
6
12
18
4
10
14
2
19
frequency
rank 2
38
0
6
23
34
41
8
16
0
84
17
45
7
24
48
3
19
37
0
69
R1 - R2 (D)
2
18
10
-4.5
-3
-8
-4
-6
17
-19
-4
-5
7
2
-9
12
1
-7.5
16
-15
15
2
5
11
13
16
7
8
2
20
9
17
6
12
18
4
10
14
2
19
D2
4
324
100
20.25
9
64
16
36
289
361
16
25
49
4
81
144
1
56.25
256
225

D = 0
D2 = 2080.5
rs = 1 - 6D2
n(n - 1)
= 1 - 6 x 2080.5
20(20 - 1)
= 1 - 12483
20 x 399
= 1 - 12483
7980
= 1 - 1.56
= - 0.56
= 0.56 (critical value = 0.45)
= 98% significant
My number for Spearmans rank is above the critical value therefore I can see that my
results are significant. My percentage significance can be worked out from the number and
shows over 95% significance.
Anomalous results:
On my graph are marked two anomalous results which are a long way from the line of best
fit. They show a far lower frequency of glasswort than expected at such high soil
compaction levels. When these two results are taken out, Spearmans rank can be repeated
to test whether or not significance is higher without them.
Sample
1
2
3
4
5
6
9
10
11
12
13
14
15
16
17
18
19
20
Distance into
soil/cm
6.9
9.9
6.1
4.2
4.7
4.4
9.2
1.2
4.0
5.7
5.8
5.9
4.5
6.6
5.6
4.2
8.2
3.3
rank 1
frequency
rank 2
15
18
13
4.5
8
6
17
1
3
10
11
12
7
14
9
4.5
16
2
38
0
6
23
34
41
0
84
17
45
7
24
48
3
19
37
0
69
13
2
5
9
11
14
2
18
7
15
6
10
16
4
8
12
2
17

71
Sample
1
2
3
4
5
6
9
10
11
12
13
14
15
16
17
18
19
20
rank 1
rank 2
R1 - R2 (D)
D2
15
18
13
4.5
8
6
17
1
3
10
11
12
7
14
9
4.5
16
2
13
2
5
9
11
14
2
18
7
15
6
10
16
4
8
12
2
17
2
16
8
-4.5
-3
-8
15
-17
-4
-5
5
2
-9
10
1
-7.5
14
-15
4
256
64
20.25
9
64
225
289
16
25
25
4
81
100
1
56.25
196
225
D = 0
D2 = 1660.5
rs = 1 - 6D2
n(n - 1)
= 1 - 6 x 1660.5
18(182 - 1)
= 1 - 9963
18 x 322
= 1 - 9963
5814
= 1 - 1.71
= - 0.71
= 0.71
= 99% significance
Significance of results is higher without the anomalies, but either with or without them, my
results have a higher than 95% significance. This disproves my null hypothesis and indicates
that my alternative hypothesis is correct.
72

Glasswort grows best at high soil compaction levels and not at all at very low levels. This is
due to the reasons stated in my introduction and prediction. I feel that the variation in
glasswort frequency at the intermediate soil compaction levels is brought about by other
factors besides soil compaction (e.g. presence of other plants and nearness to a creek).
Evaluation:
The conclusion that I have reached during this investigation is a firm one. The only results
which could be placed against it are the two marked on the graph and I feel that they are
anomalous, caused by variables which I was unable to control. The main uncontrolled variable
was the weather. It was very hot, and I think that these two samples, which were taken
some time after the others in the same section of the sample area, gave unreliable results
due to the sun having dried out and hardened the soil. There is no way of controlling this
variable, unless the investigation is carried out on a day selected for ideal weather
conditions. I was limited here as there were only two days in which to carry out the
investigation, both of which were hot and dry. I have however, made allowances for the fact
that there are anomalies in my results, and have repeated the statistics test without them,
to show the increased significance of my results when they are removed. I followed my
method as I had planned and found that it was efficient and gave me the results that I
needed to see a pattern and to be able to carry out a Spearmans rank statistics test. The
main limitation with the procedure was the lack of accuracy in measuring soil compaction.
The use of the rod left room for human error both when dropping it and when measuring how
far into the soil it had fallen and meant that although I could gain a relative value which
allowed me to see different areas of soil compaction, I could not gain values which could be
expressed using a precise unit. This was, however, the only method available to me for
taking this measurement, and although it was not strictly accurate, it gave me an indication
of the compaction of the soil, which was all that I needed to conclude that there is a
relationship between soil compaction and Glasswort.

73
Bibliography:
(1) Saltmarshes - Morphodynamics, conservation and engineering significance
J.R.L.Allen and K.Pye
Cambridge university press
1992
(2) A handguide to the sea coast
John Barrett
Treasure press
1981
(3) Effects of competition, disturbance and herbivory on Salicornia europaea
Aaron M.Ellison
Ecology vol. 68
1987
(4) Secrets of the seashore
Consultant - Derek Hall
Readers digest publication
1984
(5) www.geocities.co.uk
Type of sediments in tidal flats
Compiled by J. Casas
Image of Glasswort found at: www3.gateway.ne.jp/~natis/pages_e/genus/salicornia.htm
Total: 4/6
Moderators comments
Strand (a) the trend is identified and an appropriate statistical test was used.
Strand (b) there are biological principles that could have been used to
interpret the results. Some attempts were made in applying biological principles.
Strand (c) conclusions are supported by results and the limitation of the
method of measuring soil compaction is recognised.
74

Exemplar 10 Davids work

Investigation into the amount of polysiphonia lanosa in relation
to its position growing on the Ascophyllum nodosum
Abstract:
Polysiphonia is epiphatic to Ascophyllum nodosum which grows in eullitorial zones including

the rocky shore at Aberystwyth. This investigation shows that the distribution of the
Polysiphonia lanosa increases towards the tip of the Ascophyllum nodosum until a certain
point.
A graph to show the dry mass of Polysiphnia lanosa growing at different positions on
the Ascophyllum nodosum
2.5
dry mass of Polysiphonia lanosa /g
1.5
Series1
Poly. (Series1)
0.5
0
0
10
12
14
16
18
-0.5
position of the Polysiphonia lanosa on the Ascophyllum nodosum / proportion 16ths

75
Analysis:
Looking at the graph, there is some strong positive correlation, which shows the values
increasing until it reaches a peak in the twelfth segment. It then decreases almost forming
an inverted U shape. The results up until 12th segment suggest that the alternative
hypothesis is correct and that there could be another reason why the Polysiphonia lanosa
grows until a certain point. One possibility is that, because the Ascophyllum nodosum grows
from its tip, the Polysiphonia lanosa has not settled on to the new growth.
The U shape of the graph means that a statistical test cannot be used for the entire
results. However, in order to prove the new theory correct, the results can be separated
into two sections
positive correlation up until the peak at the twelfth segment,
negative correlation after that peak.
Unfortunately, to use Spearmans Rank one must have 7-30 pairs of measurements, but
there are only four for the negative correlation.
Appendix III.
Results:
if rs is greater than or equal to the critical value then there is a significant correlation.7
Significance level
no. of pairs
12
5%
0.591
2%
0.712
1%
0.777
The value from the figures came out as 0.9720273

The first 12 results are highly significant with less than 1% probability than the results
being due to chance.
Conclusion:
There is significant positive correlation which disproves the null hypothesis and must
therefore support the alternative hypothesis.
There are several reasons why there is much less P. lanosa past the 12th section on the
Ascolphyllum nodosum
As the A. nodosum grows from its tip outwards, the polysiphonia, as yet, has not settled on
the new growth.

Also, Ascophyllum nodosum is attached to the rocks by a holdfast, therefore can only be
moved around this point. The tip of the seaweed will be move furthest by the waves and
tide it causing lacerations as it gets beaten across the rocks. As a result of this damage,
the Polysiphonia lanosa will not grow at these points.
76

Another possibility is photoinhibition. The inhibition of photosynthesis at high intensities

of light must be taken into account in ecological studies, since the intensities typically
experienced in the surface layer of natural waters in sunny weather are in the range that
can produce photoinhibition8. However, with increasing depth and diminishing light
intensity, photoinhibition lessens and the maximum, light-saturated but not inhibited,
photosynthetic rate is achieved9 This could explain why there is a drop in the amount of the
Polysiphonia lanosa found after a certain distance. The peak of Polysiphonia lanosa growth
could be where the light intensity is great enough to reach the maximum photosynthetic
rate and therefore more will grow there.
Evaluation:
Problems with the experiment.
By picking samples by this method the Ascophyllum nodosum may not the best samples
for testing. For example, three of the samples collected did not have any Polysiphonia
lanosa on it. However, this method did rule out observer bias.
There was another epiphytic green seaweed growing on some of the Ascophyllum
nodosum samples. Some may have been scraped off and weighed with the Polysiphonia
lanosa as it was impossible to separate, therefore giving it an incorrect mass.
Digital Balance - the balance used to obtain the mass was highly sensitive and kept
changing. It measured to the nearest 100th of a gram and was highly sensitive.
All the masses obtained were averages from the 10 samples taken from the rocky shore.
Therefore, it is not possible to identify any anomalous results from one specific plant
and exclude them. I could not do each individually because there was insufficient time
to do this individually. To get more reliable results, one could measure each 10
Ascophyllum nodosum samples separately as this would not mask any anomalous results
from one single or many seaweed samples.
For a further study, one could examine the different kinds of growth of Polysiphonia
lanosa on the Ascophyllum nodosum at different points on the shore. The Polysiphonia
lanosa growing at the high water mark seemed to clump together near the top in a
different shape more like a pompom.
It was not possible to show any negative correlation which limited the veracity of the
study. This would make an interesting further investigation.
Total: 6/6
Moderators comments
Strand (a) the graph shows the trend in the data and the statistical test is
appropriate to the hypothesis.
Strand (b) biological principles are applied correctly.
Strand (c) conclusions are supported by results and limitations, with possible
modifications suggested.

77
Assessment criteria Communicating

F
Communicating
The layout of the report largely conforms to that expected of a

scientific paper. The organisation of the report produced shows
evidence of some thought and the aim of the investigation is
stated. Images, when used, are relevant to the points made.
Data are presented in graphs, tables or diagrams, which are mostly

appropriate and follow scientific conventions for presentation.
Spelling, punctuation and grammar are generally correct, some

technical terms are used appropriately and most sources used are
acknowledged in a bibliography.
The layout of the report mostly conforms to that expected of a

scientific paper with sub-headings used effectively. The aim(s) and
conclusion(s) of the investigation are stated. Images, when used,
illustrate points clearly.
Data are presented in well-chosen graphs, tables or diagrams,

which usually follow scientific conventions and mostly use SI units,
where appropriate, correctly.
Spelling, punctuation and grammar are correct, appropriate

technical terms are usually used and sources used are referred to
within a properly constructed bibliography. This is the maximum
mark for reports which exceed 3,000 words.
The layout of the report conforms to that expected of a scientific

paper with appropriate and helpful sub-headings The organisation
of the report shows evidence of thoughtful planning and the aim(s)
and conclusion(s) of the project are clearly stated and discussed.
Images illustrate the points effectively and enhance the clarity of
the report.
Data are presented effectively in graphs, tables or diagrams which

follow scientific conventions and are clearly and accurately labelled
using SI units where appropriate.
Spelling, punctuation and grammar are correct, and appropriate

technical terms are used throughout. Sources are selected and used
appropriately and are correctly and clearly referenced within a
properly constructed bibliography. The report is within the word
limit of 3,000 words.
78
Total marks
2 marks
4 marks
6 marks

Exemplar 11 Rosannas work

Investigating the effect of ethene on the ripening of fruit
Contents
Main investigation
Page Introduction
Method
Variables to consider
Results
Graph to show results
Statistical analysis
Limitations
Conclusion and modifications
References
Appendix 1 Initial plan for investigation
9, 10, 11, 12
Appendix 2 Pilot investigation and modifications
12, 13
Appendix 3 Details of Benedicts test
13

79
Appendix 4 Details of colorimeter
13, 14
Appendix 5 Raw experimental data
14, 15, 16
Appendix 6 Statistical test workings
16, 17
Aim:
To investigate the effect of Ethene on the rate of ripening of strawberries, using the rate
of starch hydrolysis into sugar as an indicating factor.
Introduction:
Ethene is an organic compound that consists of hydrogen and carbon atoms, contains a
double bond and belongs to the alkene family. The structure of ethene is: H2C === CH2 (7).
Ethene is a plant growth substance that regulates the growth and the ripening of certain
fruits. For many years ethene has played an important part in the fruit marketing business
as it allows for unripe fruit to be picked and transported globally without ripening too early.
The fruit sellers can then choose when to ripen the fruit at the time of sale with the
addition of ethene gas, which initiates the ripening process.
Naturally plants produce their own supply of ethene that, when released, serves as a
ripening signal, therefore the rate of production of ethene is higher for plants during the
ripening period. When the plant releases ethene it brings about the production of many
enzymes that contribute in differing ways to the ripening process. These enzymes include
amylases to accelerate the hydrolysis of starch into sugar along with other enzymes such as
pectinases to catalyse the degradation of pectin (1) (6) (10). It is the effect that the presence
of ethene has on the hydrolysis of starch into sugar that this experiment will investigate.
Hypothesis The presence of ethene will increase the hydrolysis of starch into sugar in
fruit.
Null Hypothesis The presence of ethene will have no significant effect on the hydrolysis
of starch into sugar.
The Spearmans Rank coefficient statistical test will be used to compare the correlation
between the rate of the ripening of the fruit and the time taken for this ripening. In order
to accept the above hypothesis the test will need to show that there is a clear correlation
between the two variables. If the statistical test shows that no clear correlations have been
produced then it could be said that the results are inconclusive and therefore, not reliable,
nor clear enough, to firmly support any conclusions.
In this investigation the effect that the ethene produced by bananas has on the ripening
process of strawberries will be investigated. Bananas produce lots of ethene when ripening
and for this reason it is not recommended to keep bananas with other fruit as they cause
them to ripen too quickly and go bad. Potassium permanganate is a substance that absorbs
80

ethene (5) (9) and will be used as a control that will eliminate ethene from one of the
containers. The rate of the hydrolysis of starch into sugar will be used to determine the
rate of ripening. This will be measured through a simple test for sugar called the Benedicts
test. When sugar is present in a solution there is a colour change from blue to a red
precipitate (see Appendix 3). The strength of this colour change will be determined through
the use of a colorimeter and this should indicate the amount of sugar present in the ripening
fruit.
Apparatus:
4 Bananas
45 strawberries (firm/unripe)
2 Sealed containers/plastic bags
Potassium Permanganate crystals
Benedicts solution
45 test tubes
46 100ml beakers
Bunsen Burner
Tripod and Gauze
Heatproof mat
Distilled water
Electronic scales
Colorimeter
45 Cuvettes
Centrifuge
Food Blender/Processor
2 pipettes with pipette droppers
Knife and tile
See Appendix 1 for a full list of the apparatus and justifications of use in the investigation.
Method:
1. 1 banana and 20 strawberries were placed in a sealed container.

2. Placed in the other container were 1 banana, 20 strawberries and 200g of potassium
permanganate crystals.
3. Containers were stored in a cool, shady area.
4. 15g of each of the 5 remaining strawberries were then blended and the sediment was
separated using centrifugation.
5. 2cm3 of the solution and 2cm3 of Benedicts solution were placed test tube in a beaker of
water and heated to test for sugar.
6. The optical density of the above solution was then determined through the use of a
colorimeter.
7. Every day for the next 5 days, 5 strawberries from each container were taken and the
above steps of 4, 5 and 6 were repeated on each strawberry and the results recorded.
Variables that need to be considered:

81
The weight of the bananas used need to be roughly equal and from the same bunch to
ensure that they are at the same stage of ripening.
The amount of strawberry tested needs to be weighed and be exactly the same for all
strawberries electronic scales can be used to ensure a weight of 15g is used in the
tests.
Use strawberries from the same pack to ensure similar levels of ripeness. However
these strawberries are still likely to vary in age and this should therefore be
considered when analysing results.
The experiments need to be carried out at the same time each day so that the period of
ripening between each set of experiments is the same.
Results:
The raw experimental results can be found in Appendix 6. Below are the average results that
have been produced as a result of the repetitions of each stage of the experiment.
82

Hours
0 hrs
24 hrs
48 hrs
72 hrs
96 hrs
Container
No ethene
Ethene
No ethene
Ethene
No ethene
Ethene
No ethene
Ethene
Average Colorimeter
reading
0.684
0.728
1.022
1.470
1.722
1.286
0.782
1.090
0.702
Statistical Analysis.
Evaluation of graph:
The two lines produced from the average results of this experiment do follow a similar
pattern and also appear to support the earlier hypothesis that ethene enhances the rate of
ripening as it is quite noticeable that the fruit surrounded by the ethene did show quicker
rates of ripening (reflected by the high optical density that represents the sugar density).
The pattern shows that for the first 24 hours into the experiment there is a steady
increase in the amount of light absorbed by the solution. An increase in the amount of light
absorbed by the solution indicates that the fruits being tested have a higher sugar content
than those previously. Solutions with higher absorbencies are the solutions containing higher
concentrations of the red Cu+ ions. The Cu+ ions come from the sugars present in the
centrifuged strawberry solution reducing the blue Cu2+ ions in the Benedicts solution and
therefore a higher concentration of these ions indicates more sugar was present in the
solution being tested. After 24 hours the OD readings start to rapidly increase for both
sets of data and the strawberries with ethene reach a peak of 1.722 and the strawberries
without ethene reach a peak of 1.470. After 48 hours both lines begin to fall, with the
ethene line dropping most rapidly. The reason for the line that showed quickest ripening to
also show the quickest fall of sugar content is largely due to the development of mould on,
and, inside the fruit (see below). It should be expected that the fruits that ripen most
quickly would also go off most quickly. After 96 hour the fruits without ethene maintained a
higher sugar content than initially started with, whereas the fruits that had been
surrounded by ethene had sugar contents only marginally above that of which they started
at.
Spearmans rank correlation:
(See Appendix 7 for workings and discussion) The results of this statistical test produced a
Spearmans rank correlation coefficient of 0.35. In order for the calculated coefficient to
be accepted it has to be within a 5% significance level that varies depending on the number
of pairs of variables being used. The 5% significance level when 10 pairs of variables have
been used is 0.65 and above. Therefore the calculated coefficient of 0.35 from the results
is not high enough to accept a firm correlation and therefore supports the acceptance of
the null hypothesis that the presence of ethene will have no significant effect on the
hydrolysis of starch into sugar.

83
The trouble with the results produced is that although the amount of time continues to rise,
the measured sugar content reaches a peak in the middle of the experiment and then starts
to drop off towards the end. This creates problems with the ranking of the results and
therefore results in large differences between the rankings of the variables being
calculated. These large differences could explain the relatively low coefficient produced and
therefore, why a clear correlation could not be detected. For this reason the results from
the Spearmans rank test cannot be entirely conclusive with the acceptance of the null
hypothesis.
Anomalous results:
Some slightly anomalous results were produced in this experiment such as the colorimeter
reading of 1.38 on the strawberries without ethene after 24 hours, and also the reading of
0.43 on the strawberries after 96 hours. It is almost certain that the strawberries will not
all be at the same level of ripeness and this could have caused some of the anomalous
recordings. The riper strawberries will obviously have a higher sugar level than the others
and this explains the high reading of 1.38. Later on in the experiment there was a problem
with mould growing on and into the fruit. As the mould grew into the strawberries it had the
effect of changing the colour of the fruit to brown, in turn this had the effect of lowering
the colorimeter reading. This would therefore explain why later on in the experiment all of
the colorimeter readings have lowered, as reflected in the line graph, due to the growth of
mould on and into the remaining strawberries.
Limitations of the experimental procedure:
84
The ages of the strawberries were unknown and more than likely to be different. Ideally
unripe strawberries of exactly the same age would have been purchased, as these would
have produced the clearest results.
The fact that the strawberries differed in size is also a factor that will have had an
impact on the results as the surface area to volume of the fruit ratio would vary
considerably and therefore the impact of the ethene gas on the fruit is likely to have
had more of an effect on the smaller strawberries.
The distance of each individual strawberry from the banana in the container varied.
Although the ethene gas produced by the banana would have diffused around the
container it was evident that the strawberries immediately next to, or touching, the
banana were showing clear signs of ripening more quickly than those further away.
Ideally it would have been preferable to place each of the strawberries at an equal
distance from the banana to ensure that the experiment is a fair test.

Conclusion:
In conclusion, bearing in mind the results of the statistical test and the graph, it can be said
that the results from this investigation were not conclusive enough to firmly accept or
reject either the hypothesis or the null hypothesis. Although the results from the graph
seem to clearly support the hypothesis that the presence of ethene increases the hydrolysis
of starch into sugar in fruit, the results from the Spearmans rank correlation show that the
results were not conclusive enough to show clear correlations between the overall rates of
ripening and the time taken. As mentioned in the statistical analysis the results from this
investigation were not ideal for use with the Spearmans rank correlation and therefore the
results from the graph are more accurate to deduce conclusions from. Taking this into
consideration the results produced, which are best presented in the graph, do support,
although not conclusively, the hypothesis that ethene does increase the rate of sugar
production in strawberries.
Modifications and suggestions for further work:
Ideally another container that contained only strawberries would be used, as this would
monitor their natural rate of ripening. This would provide a basis to compare the results
with and also help determine if other substances released by the bananas were causing
ripening.
Also, all of the fruits used would have been surface sterilised, as this would prevent
contamination of the results by ensuring that any rates of ripening are only to do with the
substances inside the containers.
The effect of using unripe and ripe bananas to see which ones helped to ripen the fruit more
quickly could also have been investigated. During the background research to this
investigation it was discovered that fruits produce more ethene when stressed/wounded so
it would have therefore been interesting to investigate the effect that damaged bananas
had on the ripening of the strawberries compared to the effect of normal, healthy bananas.
References:
1. http://plantphys.info/plants_human/frruitgrowripe.html
2. Biology for life, M.B.V. Roberts, published by Nelson 1986, Food and Diet, Pg.56. ISBN
0-17-448096-2
3. Practical Skills in Biology, Jones, Reed and Weyers, published by Longman group UK
1994.ISBN 0-582-06699-9
4. http://images.google.com/images?hl=en&lr=&ie=UTF-8&oe=UTF-8&q=centrifuge
5. http:www.ars.usda.gov/is/kids/ plants/story7.html
6. http:www.ezthemes.com/previews/ b/bananas.jpg
7. http://www.tiscali.co.uk/reference/encycolaedia/hutchinson.html
8. http://www.sciencenet.org.uk/database /bio/plants/otherplant/b00707d.html
9. http://syi.hkcampus.net/syi-kc/expll.html
10. http://www.saps.plantsci.cam.ac.uk/osmoweb/ethenemenu.htm
Appendix 1:
Plan Investigating the effects of ethene on fruit ripening

85
Introduction:
Ethene is an organic compound that consists of hydrogen and carbon atoms, contains a
double bond and belongs to the alkene family. The structure of ethene is: H2C === CH2.
Ethene is a plant growth substance that regulates the growth and the ripening of certain
fruits. For many years ethene has played an important part in the fruit marketing business
as it allows for unripe fruit to be picked and transported globally without ripening too early.
The fruit sellers can then choose when to ripen the fruit at the time of sale with the
addition of ethene gas, which initiates the ripening process.
Naturally plants produce their own supply of ethene that, when released, serves as a
ripening signal, therefore the rate of production of ethene is higher for plants during the
ripening period. When the plant releases ethene it brings about the production of many
enzymes that contribute in differing ways to the ripening process. These enzymes include
amylases to accelerate the hydrolysis of starch into sugar along with other enzymes such as
pectinases to catalyse the degradation of pectin. It is the effect that the presence of
ethene has on the hydrolysis of starch into sugar that this experiment will investigate.
Hypothesis The presence of ethene will increase the hydrolysis of starch into sugar in
fruit.
Null Hypothesis The presence of ethene will have no significant effect on the hydrolysis
of starch into sugar.
Statistical test - I will use the Spearmans Rank coefficient statistical test to compare
the correlation between the rate of the ripening of the fruit and the time
taken for this ripening. In order to accept the above hypothesis the test will need
to show that there is a clear correlation between the two variables of time and rate of
ripening. Clear correlations will show that the results produced from this investigation are
significant enough for conclusions to be deduced from. However, if the statistical test
shows that no clear correlations have been produced then it could be said that the results
are inconclusive and therefore, not reliable, nor clear enough, to firmly support any
conclusions.
86

In this investigation the effect that the ethene produced by bananas has on the ripening of
tomatoes will be investigated. Bananas tend to produce lots of ethene when ripening and for
this reason it is not recommended to keep bananas with other fruit as they cause the other
fruits to ripen too quickly and go bad. Potassium permanganate is a substance that absorbs
ethene and will be used as a control that will eliminate ethene from the containers. The rate
of the hydrolysis of starch into sugar will be used to determine the rate of ripening. The
rate of this hydrolysis will be measured through a simple test for sugar called the Benedicts
test. When sugar is present in a solution there is a colour change from blue to a red
precipitate. The strength of this colour change will be determined through the use of a
calorimeter and this should indicate the amount of sugar present in the ripening fruit.
Apparatus
4 Bananas
45 tomatoes (firm/unripe)
2 Sealed containers/plastic bags

Potassium Permanganate crystals
Benedicts solution
45 test tubes
46 100ml beakers
Bunsen Burner
Tripod and Gauze
Heatproof mat
Distilled water
Electronic scales
Colorimeter
45 Cuvettes
Food Blender/Processor
2 pipettes with pipette droppers
Knife and tile
Justification
Fruit known to produce large quantities of ethene gas
Quite easy to obtain unripe tomatoes therefore good
for noticeable differences between ripe and unripe
fruit.
Airtight do not allow other gases that may affect
the results of the experiment enter.
Crystals that absorb ethene gas (5).
Solution used to test for sugar.
For use in the Benedicts test.
To hold the water that is heated in the Benedicts test
and also to hold the blended strawberry and distilled
water solution.
To heat water in the Benedicts test.
To safely hold and support the beaker of water being
heated during the Benedicts Test.
Safety when using Bunsen burner
For dilution and rinsing.
For accuracy in measurement when weighing the
strawberries.
Used to test optical density of a solution.
Holders for the solution when placed inside the
colorimeter.
To liquidise the strawberries and distilled water.
One to accurately measure the Benedicts solution
and the other to measure the centrifuged strawberry
solution.
To cut the strawberries safely ready for weighing.
Method:
1. In a sealed container place 2 bananas and 20 of the unripe tomatoes.

2. In the other sealed container place 2 bananas, 20 of the unripe tomatoes and 200g of
potassium permanganate crystals.
3. Ensure that both containers are firmly sealed and store in cool, shady area i.e. a
cupboard.
4. The five remaining tomatoes need to be tested for their sugar content.
5. First Take one tomato, remove the green stalk and using a knife and electronic scales
cut and weigh the strawberry to 15g.

87
6. Place the tomato in a blender with 50ml of distilled water and blend until the solution is
smooth and all large pieces are removed.
7. Pour the blended solution into a beaker and then rinse the inside of the blender with
25ml of distilled water and pour this into the same beaker.
8. Using a funnel and filter paper, filter the blended solution into a beaker.
9. Pipette 2ml of the filtered solution into a boiling tube and using a separate pipette,
pipette 2ml of Benedicts solution into the same tube, perform the Benedicts test (see
Appendix 3).
10. Now with the solution that was produced by the Benedicts test the colorimeter (see
Appendix 4) can be used to determine the optical density of the solution. This will give
an indication to the concentration of sugar present in the solution. The units for the
colorimeter reading OD for optical density.
11. Every day for the next four days take five tomatoes from each container and follow the
above process, recording the results in a table like the one below.

Container with Ethene present
After 24 hours
Colorimeter
Reading (OD)
1
2
3
4
5
Average
Variables that need to be considered:
The weight of the bananas in each container need to be roughly equal and from the same
bunch to ensure that they are at the same stage of ripening and releasing similar
amounts of ethene.
The amount of tomato tested needs to be weighed and be exactly the same for all
tomatoes electronic scales can be used to ensure a weight of 15g is used in the tests.
Ideally all of the tomatoes will be at exactly the same stage of ripening at he beginning
of the experiment and it is therefore important to use tomatoes from as few stalks as
possible. However it will not be possible to get 45 tomatoes from one stalk and so the
tomatoes will therefore be of varying age.
Ideally the experiments need to be carried out at the same time each day so that the
period of ripening between each set of experiments is the same.
Risk Assessment:
There is only a small amount of chemical risk involved with this experiment and that involves
the use of the potassium permanganate and the Benedicts solution. Both of which are
harmful on consumption and can be irritants. It is therefore important that protective
safety items and clothing is worn when dealing with these substances. This includes safety
coats, protective eyewear and gloves. If there are any spillages take care when cleaning up
and avoid placing any objects near mouth.
88

The experiment also involves the use of a Bunsen burner and therefore the necessary
precautions of heatproof mats and the protective clothing mentioned above must be worn at
all times to reduce the risk of burns.
The experiment involves the use of lots of fragile glassware and so therefore great care
needs to be taken at all time to avoid any breakages. If a breakage does occur, warn other
nearby, use a dustpan and brush clear up the broken glass as this will avoid cuts and place
the broken glass in the special glass bins in the lab.
Modifications:
Modifications have been made to this initial plan with the respect that strawberries will be
used instead of tomatoes. The reason for this is because strawberries show a more
noticeable change in sugar content from the unripe to ripe stage. Tomatoes, even when fully
ripened, are not as sweet and therefore it would be slightly harder to detect changes in the
sugar level. Appendix 2 below shows the pilot investigation involving strawberries and
further modifications that arose as a result of this.
Appendix 2.
Pilot Investigation:
In order to test the method of this investigation a trial method was necessary to see if any
modifications would need to be made to improve the accuracy of the results. The following
process is the original process that was decided upon and is the same process that is to be
followed for each individual strawberry (along with the modifications below), taking care to
wash all apparatus before the use of a new strawberry.
a) Take one strawberry and carefully remove the green stalk.
b) Using a knife and electronic scales cut and weigh the strawberry to 15g.
c) Place the strawberry in a blender with 50ml of distilled water and blend until the
solution is smooth and all large pieces are removed.
d) Pour the blended solution into a beaker and then rinse the inside of the blender with
25ml of distilled water and pour this into the same beaker.
e) Using a funnel and filter paper, filter the blended solution into a beaker.
f) Pipette 2ml of the filtered solution into a boiling tube and using a separate pipette,
pipette 2ml of Benedicts solution into the same tube, perform the Benedicts test (see
Appendix 3).
g) Now with the solution that was produced by the Benedicts test the colorimeter (see
Appendix 4) can be used to determine the optical density of the solution. This will give
an indication to the concentration of sugar present in the solution.
Modifications:
Whilst doing the trial test it was realised that some modifications would be needed. The
blended strawberry solution was too substantial too pass through the filter paper. Tissue
paper was then used but this too proved impermeable for the solution. In order to
successfully separate the particles from the solution a process called centrifugation was
decided upon. Step e) needs to be replaced with
e) Measure 5ml of the solution and place in a centrifuge tube and centrifuge the solution.

89
Also, after the Benedicts test, the solution that was left for use in the colorimeter was too
dense to produce any readings. After some trial and error it was decided that 20ml of
distilled water was an appropriate amount to dilute the solution by. Therefore in the main
investigation g) needs to be replaced by
g) Dilute the solution from the Benedicts test with 20ml of distilled water and use
thecolorimeter (see Appendix 4) to determine the optical density of this solution. This
will give an indication to the concentration of sugar present in the solution.
Appendix 3: Benedicts test.
Benedicts solution contains blue copper sulphate Cu2+ ions that when heated are reduced by
sugar to the red Cu+ of copper (I) oxide (2). Sugars that do this include glucose and fructose
and lactose and are called reducing sugars. When there is insufficient sugar present only the
granules are seen through the solution and it may appear green, slightly more precipitate
gives a yellow appearance and therefore different colours can be used to determine the
amount of sugar present in the solution being tested.
The test 1) The food needs to be in liquid form use the solution present after the
centrifugation.
2) Pour 2 cm3 of the food solution into a test tube.
3) Add 2 cm3 of the Benedicts solution to the test tube and shake.
4) Boil some water in a beaker over a Bunsen burner.
5) Put the test tube in the beaker of boiling water and leave it there for a minute or two and
note the colour change that occurs.
Appendix 4: Colorimeter.
This process can be used with solutions where the substance of interest is highly coloured
and present as the major constituent i.e. the red Cu+ of the copper (1) sulphate.
How to use a colorimeter 1) Switch on and stabilise allowing at least 5 min for the lamp to
warm up.
2) Choose a complementary filter to the colour being measured. The colour we are measuring
is red as it absorbs light within the blue/green regions of the spectrum, therefore a blue
filter should be used.
3) Set the scale zero using distilled water as the blank solution.
4) Adjust the sensitivity to give a reasonable deflection on the galvanometer/readout
device.
5) Fill the cuvette to the appropriate level and place in the colorimeter ensuring the
outside of the cuvette is dry and that the correct side of the cuvette is facing the correct
way. Record the reading in a results table; the units are OD for optical density.
6) Clean out the cuvette between samples with distilled water.
90

Appendix 5: Raw Data produced from the experiment.

Results Day 1
Unripe strawberries beginning of
experiment
After 0 hours
1
2
3
4
5
Average
Colorimeter
Reading (OD)
0.73
0.70
0.65
0.59
0.75
0.684
Day 2
After 24 hours

1
2
3
4
5
Average

After 24 hours
1
2
3
4
5
Average
Colorimeter
Reading (OD)
0.87
0.77
0.71
1.38
0.91
0.728
Colorimeter
Reading (OD)
1.04
0.92
0.84
0.95
1.36
1.022

91
Day 3
After 48 hours
1
2
3
4
5
Average

After 48 hours.
1
2
3
4
5
Average
Colorimeter
Reading (OD)
1.29
1.44
1.54
1.58
1.50
1.470
Colorimeter
Reading (OD)
1.69
1.89
1.49
1.68
1.86
1.722
Day 4
Strawberries without Ethene
present
After 72 hours
1
2
3
4
5
Average

After 72 hours
Average
92
1
2
3
4
5
Colorimeter
Reading
(OD)
0.83
1.58
1.14
1.28
1.60
1.286
Colorimeter
Reading (OD)
0.96
0.60
0.98
0.74
0.63
0.782

Day 5
After 96 hours
1
2
3
4
5
Average

After 96 hours
1
2
3
4
5
Average
Colorimeter
Reading (OD)
1.75
0.99
0.98
1.12
0.61
1.090
Colorimeter
Reading (OD)
0.94
0.72
0.43
0.74
0.68
0.702
Appendix 6: Statistical test.
As mentioned earlier in the plan, the statistical test that will be used in this investigation is
the Spearmans Rank coefficient and this will be used to determine the strength of the
correlation between the rates of ripening of the fruit and the time taken. In order to
accept the hypothesis the Spearmans rank correlation coefficient needs to show that the
results have 5% or less probability that chance alone could have produced the correlation.
The Spearmans rank correlation coefficient is slightly less likely to reveal the existence of
a correlation than other more detailed correlation tests such as the Pearson productmoment correlation coefficient. However, if a clear correlation is present within the data
then this will be shown from the results of the Spearmans rank correlation coefficient, and
will be supportive of any conclusions drawn.
Spearmans Rank Correlation
Coefficient: rs = 1 - 6d2
(n3-n)
Where: rs = Spearmans rank coefficient
d = difference between ranks
n = (hours)
number ofRank
pairs of variables
Time
Colorimeter
0
24
48
72
96
0
24
48
72
96
9.5
7.5
5.5
3.5
1.5
9.5
7.5
5.5
3.5
1.5
reading (OD)
0.634
0.728
1.470
1.286
1.090
0.684
1.022
1.722
0.782
0.702
Rank
d2
9.5
7
2
3
5
9.5
4
1
6
8
0
0.5
3.5
0.5
3.5
0
3.5
4.5
2.5
6.5
0
0.25
12.25
0.25
12.25
0
12.25
20.25
6.25
42.25

93
Using - rs = 1 - 6d2
(n3-n) when n = 10
rs = 1 6 118.25
1000-10
rs = 1 709.5 = 0.35
1090
So rs = 0.35.
The critical value for n = 10 at the 5% significance level 0.65 and above, therefore on the
basis of this statistical close there is approximately a 35% probability that chance alone
produced the correlation and so the hypothesis that the presence of ethene will increase
the hydrolysis of starch into sugar in fruit has to be rejected and the null hypothesis that
the presence of ethene will have no significant effect on the hydrolysis of starch into sugar
is accepted.
The problem with using the Spearmans Rank correlation coefficient for the results of this
experiment is that the number of n is quite low at 10; ideally a minimum of 15 pairs of
variables would be used. The low number of variables means that there is less chance of
proving that a correlation exists with this test even though a correlation may be present.
Total: 5/6
Moderators comments
Strand (a) the layout is in the style of a scientific paper with subheadings, but
the images did not illustrate points clearly. An abstract was very helpful.
Strand (b) the graph could have had the points joined by a ruler and the data
could have been combined in one table.
Strand (c) the bibliography was well-constructed and referenced in the text. To
conform to the word limit there is an extensive range of appendices. With a more
concise style of writing, some information could have been included in the main
body of the report.
produce the graph by electronic means and improve the table of raw data
use a more concise style of writing.
94

Exemplar 12 Jans work

Investigating the effect of organic pollution
on the dissolved oxygen levels in the Ouse river
Abstract
This study looks at the localised effect of a point source continuous discharge of organic
waste on a freshwater ecosystem, focussing closely on changes in Biological Oxygen Demand
(BOD) and dissolved oxygen content. I predicted a rise in BOD around the outfall and a fall
in dissolved oxygen content shortly after which would then gradually recover. I found a
strong negative correlation between BOD and dissolved oxygen levels which supported my
hypothesis. It was evident from the results that other factors were affecting the dissolved
oxygen levels. I concluded that further work with a revised approach would yield more
conclusive results.
Rationale
I decided to investigate freshwater pollution as the river Ouse that flows through
Buckingham, where I live, would be perfect for that kind of investigation. It is a rural
freshwater ecosystem with abundant wildlife and is subjected to various sources of
pollution. I settled on investigating a change in BOD and dissolved oxygen as these can be
tested for in the lab and would be a direct result of the nearby sewage treatment plant.
Effects of pollution in freshwater ecosystems

Freshwater environments support a diverse range of insect life, fish, birds and amphibians.
This is partly because the conditions in this environment, like temperature, flow rate, ph and
oxygen content, can change quickly and fluctuate over short distances. Insects and aquatic
plants have evolved to survive in particular conditions within the freshwater ecosystem and
have a low tolerance for changes in these conditions. Larger animals have a higher tolerance
for environmental change but still depend on the insects and plants for survival.
Pollution can be divided into that which is directly toxic to the organisms and that which
affects the physical environment in which those organisms live, affecting them indirectly.
Where there is a point source of pollution with a continuous discharge like an industrial
outfall or a sewage works, the chemical effects on the water gradually lessen further
downstream as the pollutant is broken down by micro organisms and becomes less
concentrated. This can create a biological succession of species with those most tolerant to
the effects of the pollution closest to the outfall.
One of the principle effects of organic pollution from a sewage works is to cause a build up
organic matter causing a bloom of aerobic microbes which break down the organic waste
using up the dissolved oxygen in the water. This creates a high Biological Oxygen Demand
which leads to a drop in the dissolved oxygen level. This breakdown is a relatively slow
process so the lowest level of dissolved oxygen is usually some way downstream of the
outfall. This is shown in diagram (a). The amount of drop in the dissolved oxygen level
depends on the BOD; defined as the amount of oxygen consumed by a known volume of
effluent over five days in the dark at 20C, and the rate of recovery depends on other
environmental factors like temperature, extent of re-aeration, numbers and types of
microbes and total organic load of the river. A high drop in oxygen levels can make the water
anoxic, killing most life from the lower trophic levels up.

95
Diagram from Biology of Habitats series, The Biology of Streams and Rivers
Paul . S . Giller and Bjorn Malmqvist. Oxford University Press
These diagrams represent the

longitudinal zonation established
downstream of a continuous
effluent discharge (a) and (b) are
physical and chemical changes;
(c) changes in micro organisms
and plants; (d) changes in larger
organisms.
[Labeled aerial photograph of effluent outfall and Sewage

Treatment Works omitted by Edexcel]
Aerial Photograph of the study site -The Ouse River just west of Buckingham, Bucks, UK
Hypothesis
As the aerobic microbes bloom on the effluent from the sewage works the BOD will increase
and they will use up the dissolved oxygen in the water. This will then gradually replenish
when the effluent is broken down, the microbes die back, and the BOD falls. The distance
of the point of lowest oxygen from the outfall will depend on the rate of flow. The rate of
recovery will depend on the quantity of photosynthetic plants in the water and effects of
flow rate and churning. The more plant life, the more churning and the slower the current,
the closer to the outfall normal oxygen levels will be restored. BOD will be highest closest
to the outfall and decrease downstream from the outfall.
96

Null Hypothesis
Any observed changes in dissolved oxygen level do not correlate with the position of the
sewage outfall. There is no fall in oxygen levels downstream from the outfall. BOD does not
decrease the further downstream from the outfall.
[Relevant OS map of area omitted by Edexcel]
Method
I took ten water samples from the river at one hundred metre intervals. I chose intervals
of this size based on the rate of flow; they are far enough apart to show a change but not
too far as to miss showing the change in BOD. I filled screw top pots by immersing them
slowly into a bucket sample from the river and screwing on the lids underwater so no air was
trapped inside. The samples were taken back to the lab to test for oxygen levels. The
samples must be tested soon after they are taken or the microbes begin to break down too
much of the effluent in the sample, using up oxygen, this gives inaccurate BOD readings.
They must also be kept in the dark to prevent photosynthetic algae from raising the oxygen
content. I had originally planned to use a dissolved oxygen meter with an electrolytic sensor
which would have given accurate results quickly. However none of the colleges meters were
functioning so I had to resort to the more time consuming Winkler test. This meant this
first set of samples was lost as I had to prepare the solutions for the titration.
Taking a second set of samples meant I didnt have time to incubate them for the full five
days, I incubated them for only two days, I expect this may produce inaccuracys in the final
BOD result as the shorter incubation time means a larger percentage error.
Each sample was divided into two sub-samples for the Winkler titration, one to measure the
initial dissolved oxygen level and one for comparison after incubation, the difference being
the BOD.
Variables to control
Conditions can change quickly throughout the day affecting the quantity of dissolved oxygen
considerably. To allow for accurate comparison all the water samples must be taken at the
same time of day. Samples must be kept air-tight from the moment they are taken until they
are tested for oxygen to avoid any extra oxygen dissolving into the sample.
Temperature of the samples when tested must be the same.

97
Dependant variables
The BOD will depend on the number of microbes, sewage fungus and other aquatic plants in
the area of the outfall, the more life the higher the demand for oxygen.
The level of dissolved oxygen depends on the aeration of the water by photosynthetic plants
and churning on the surface allowing oxygen to dissolve more readily.
The change in dissolved oxygen depends on the BOD, the higher the demand for oxygen the
greater the fall in oxygen levels downstream.
Materials and apparatus for the Winkler Titration

Stage 1
Manganese(II) sulphate 48% (IRRITANT)

Potassium iodide 15% in potassium hydroxide 70% (CORROSIVE)
Sample bottles of river water
Latex gloves
Safety glasses
Stage 2
Sulphuric acid 50% (CORROSIVE)

Sodium thiosulphate 0.31%
Starch solution 0.1%
Burette
Burette stand
10ml pipette and pipette filler
100ml conical flasks
Filter paper
thermometer
Latex gloves
Safety glasses
Method for the Winkler Titration

Stage 1
I divided each sample into sub-samples (a) and (b). I added 0.1ml of manganese(II) sulphate
solution to sub-samples (a), and mixed carefully, trying not to let in air. I then added 0.2ml of
alkaline potassium iodide, and mixed in the same way. A brown precipitate formed. Then the
(a) sub-samples were refrigerated and the (b) sub-samples were stored in the dark at room
temperature for two days.
Stage 2
At this point I performed stage one on sub-samples (b). I then added 0.3ml sulphuric acid to
all samples, and mixed. after the samples had stood for three minutes the precipitate had
dissolved. I filled the burette with thiosulphate solution and adjusted to zero then
transferred 10ml of the first subsample to a conical flask, and added a few drops of starch
solution. The sub sample turned blue. I titrated the sub sample with thiosulphate and used a
white tile under the flask to judge when the solution had turned clear. I repeated the
titration with another 10ml of the same sub-sample. I repeated the above with each subsample.
98

Each 1ml of thiosulphate titre was equivalent to 0.1mg of oxygen in the 10ml sub-sample. Thus
1ml of thiosulphate is equivalent to 1mg oxygen per 100ml fresh water.
The BOD is the difference between the dissolved oxygen of sub-sample (a) and that of subsample (b).
Conclusion
At the first glance the results seem to show a negative correlation and the main fluctuation in
BOD and dissolved oxygen is in the correct position to be a result of the outfall. As the BOD
rises just past the outfall the dissolved oxygen level in the river falls and the lag in response
seems to fit with the rate of flow which increases by the outfall. After this, further
downstream the readings begin to fluctuate but the negative correlation appears to continue.
Because the negative correlation continues I am led to believe these fluctuations could be
caused by other factors which affect dissolved oxygen rather than a fault in my readings.
These factors could include distribution of photosynthetic plants and isolated incidences
where churning water mixes in more oxygen. When the results were put into a Least Squares
linear correlation r = -0.683 and p = 0.029 proving this correlation with 97% surety. Y=1.012
X + 51.479 showing a shallow negative correlation. The direct effect of BOD on dissolved
oxygen is lessened by the slow rate that the microbes break down the effluent.

99
Evaluation
Human error
Slight discrepancy when judging the turning point in the titration could have caused
significant errors in such small samples. Letting in varying amounts of air when mixing the
chemicals for the titration was unavoidable with the method I used and could have thrown
off the results.
Problems in Sampling
Even within the width of the river there can be variations in dissolved oxygen and the
quantity of microbes. Areas where the flow doubles back can create isolated pockets with
different conditions than the rest of the river. When sampling I tried to collect the water
from as close to the centre of the river as possible to give a reading of average conditions
but this often means using sample pots on a pole which is clumsy and tends to create
churning which mixes the water with air. In areas where the river is broadens and deepens
effluent becomes more dilute decreasing its effect.
Further work
If I were to repeat the experiment the method could be revised for greater accuracy in the
results. Airtight sample pots using a siphon system to draw in water would eliminate the
problems of samples mixing with air. They could be attached to a pole to take samples from
the centre of the river. A portable electric oxygen meter could be used to take first set of
readings on site, before the oxygen levels are altered by aerobic microbes and
photosynthetic algae.
Smaller samples for titration give a larger margin of error, if I had to use the Winkler
titration larger samples would give better results. However my preference would be to use a
electronic oxygen meter as the samples would not suffer from mixing with air when the
chemicals are added for the titration.
Further work into the effects of the organic pollution could include surveys investigating
how animal and plant populations vary along the river to find which species are most
resistant to a fall in dissolved oxygen and tests for other chemical changes in the water
quality like acidity. The direct effect of algal blooms, triggered by fertiliser run off from
nearby farms, could also be investigated. As well as the secondary effects from the
subsequent anaerobic conditions on the plant and animal life in and around the water.
2054 Words
Bibliography
100
Biology of Habitats series, The Biology of Streams and Rivers. Paul . S . Giller and Bjorn
Malmqvist. Oxford University Press
Practical Ecology series, freshwater Studies
John H.R Gee
www.getmapping.plc
www.fishace.demon.co.uk
http://lorien.ncl.ac.uk/ming/polmon/eutrophication.pdf

Appendix 1
Planning:
I decided to investigate some form of freshwater pollution as the river Ouse that flows
through Buckingham Where I live would be perfect for that kind of investigation. I
searched the library and the internet for information on freshwater ecosystems and the
way they can be polluted. I settled on a change in BOD and dissolved oxygen as a factor
which could be tested in the lab and would be a direct result of the nearby sewage
treatment plant.
I went to the site and located the sewage outfall. I took practice samples, one from above
and one from just below the outfall, and took them back to the lab as a preliminary test to
check the range of decrease in dissolved oxygen. I didnt have time to incubate the test
samples for five days to find the BOD before starting the actual investigation.
Background:
Fast flowing water, because of its turbulence, will tend to be saturated with air, or nearly
so, in contrast to static water where the oxygen supply may become depleted, particularly
during winter.
The chemical composition
Changes in nitrate levels resulting from organic pollution may affect the ecology of a river or
stream directly. The use of nitrate fertilisers, or the break down of sewage, silage can lead
to an algal bloom which may cause changes to the effective substrate by trapping and
creating organic matter altering the ecology and oxygen levels.
Close to a point source of continuous organic pollution like the outfall from a sewage works a
biological succession is usually established with those species most adapt to the anaerobic
conditions created by the pollution nearest the outfall and those susceptible to lack of
oxygen dying out or migrating away from the source.
Method:
I decided to use a dissolved oxygen meter with an electrolytic sensor as this would give
accurate results quickly so I could take many samples and get a detailed picture of how
these environmental factors changed along the course of the river.

101
I decided to take a series of ten water samples from the river at one hundred metre
intervals. I chose intervals of this size based on the rate of flow; they are far enough apart
to show a change but not too far as to miss showing the change in BOD. I decided to take
two readings from above the outfall as a comparison to the polluted water. The samples
would be taken in plastic screw top pots with an airtight seal. I had devised a method of
extending my reach closer to the centre of the river so results are not thrown by isolated
pockets with different conditions created by the bank. This involved a pole with a hole
drilled on one end for a loop of thick string which could be wrapped securely round the rim
of the sample pot. The pot could then reach away from the bank and be removed afterwards.
However this method when tested proved too difficult to bring back the pots without
leaving an air gap at the top which would dissolve into the sample in transport. the sample
pots needed to be closed while completely submerged to avoid this problem of trapped
bubbles. A more complex sampling container with a siphon system could be constructed for
total surety tat the samples do not mix with air but for the purposes of my experiment a
simpler technique will give adequate results. I would use a bucket to take enough water from
each test site to submerge the sample pots.
Variables to control:
Conditions can change quickly throughout the day affecting the quantity of dissolved oxygen
considerably. To allow for accurate comparison all the water samples must be taken at the
same time of day. Samples must be kept air-tight from the moment they are taken until they
are tested for oxygen to avoid any extra oxygen dissolving into the sample. Temperature of
the samples when tested must be the same.
Dependant variables:
The BOD will depend on the number of microbes, sewage fungus and other aquatic plants in
the area of the outfall, the more life the higher the demand for oxygen. The level of
dissolved oxygen depends on the BOD, the higher the demand for oxygen the greater the
fall in oxygen levels downstream.
Risk assessment for taking samples:
The largest danger here involves getting wet, the current is not strong enough to be
dangerous. Swans should be given there space and treated with respect. The waters pretty
cold though, bring gloves and a hat and Wellington boots.
Risk assessment for the Winkler test:
Gloves, lab coats and goggles must be warn to protect against irritant or corrosive chemicals
and the test should be performed standing to reduce the risk of chemicals being spilled onto
your lap. Care must be taken when handling fragile glassware like pipettes, burettes and
thermometers.
Total: 5/6
102

Moderators comments
Strand (a) the layout was in the style of a scientific paper with helpful
subheadings. The images gave clarity to the report.
Strand (b) the graphs were effective, but the raw data should have been
tabulated either in the main text or as an appendix.
Strand (c) there are not many sources used and the bibliography lacks some
details. The sources were not referenced in the text.
tabulate the data
reference sources and give source details.

103
Exemplar 13 (maximum mark) Katharinas work

BIOLOGY COURSEWORK (A2)
An investigation into the effect of sodium chloride on the
action of antibiotics on bacterial growth.
Introduction:
People with the genetic condition cystic fibrosis (CF) are more prone to lung and airway
infections than non-sufferers.1 Airway secretions that include antimicrobial proteins such as
lactoferrin and lysozyme normally protect the airways and lungs from airborne bacteria
which land in the airway surface liquid (mucus). However, the effectiveness of these
naturally-occurring antimicrobial proteins, so-called natural antibiotics, is often reduced in
CF patients, and this is thought to be due to a higher than normal concentration of sodium
chloride.2 I plan to investigate whether the concentration of salt has an effect on medicinal
antibiotics by measuring the area of bacterial growth inhibition around antibiotic discs on
agar containing salt at different concentrations.
Figure 1
Scanning electron micrograph photograph showing
bacteria on the surface of airway epithelium cells.
Cilia are on the cell surface as well as attached
bacteria. 3
Hypothesis:
The antibiotic streptomycin will inhibit the growth of the bacterium E. coli to a greater
extent in lower salt concentrations. The higher the salt concentration, the smaller the
effect of inhibition.
Several variables will affect the experiment:
Temperature will affect the rate and extent of bacterial growth on agar plates. The plates
will therefore be incubated at a constant temperature (40 oC). This will ensure that
fluctuations in air temperature will not affect bacterial growth during experiments
conducted over a long period.
The incubation time will affect the extent of growth. The diameter of the area of inhibition
will be measured at the same time interval for each plate (twenty-four hours after each
plate is inoculated).
The volume of bacterial broth placed on the plate will affect the rate and extent of
bacterial growth. The volume used will be kept to three drops for each plate (using a teat
pipette).
Safety considerations: (see also risk assessment, Appendix A)
Safety spectacles were worn when using dangerous equipment such as Bunsen burners,
bacterial broth, and ethanol.
A lab coat was worn when using bacteria.
The bench was wiped with disinfectant after practical work was completed.
104

Care was taken when using boiling water.

Ethanol was kept covered and not used in close proximity to a naked flame.
Plastic petri dishes containing bacterial cultures were chemically sterilised before being
disposed of. Other contaminated equipment was autoclaved.
Method
After appropriate preliminary work (Appendix B) the following method was devised.
Preparation of agar plates with varying salt concentrations:
Standard solutions of varying concentrations of sodium chloride were made. The appropriate
mass of salt (measured using an electronic balance, accurate to one hundreth of a gram) was
dissolved in distilled water in a 100 cm3 volumetric flask, ensuring all vessels containing salt
were rinsed with distilled water into the flask. (See Appendix C). Solutions of the following
concentrations were made: 0.0 M, 0.1 M, 0.2 M, 0.3 M, 0.4 M, 0.5 M, 0.6 M, 0.7 M, 0.8 M, 0.9
M, and 1.0 M. The investigation was later extended, and stronger concentrations were used
in addition (1.5 M, 2.0 M, 2.5 M, 3.0 M). Using a graduated pipette and pipette filler, samples
with a volume of 5 cm3 were taken from each of the standard salt solutions and put into
McCartney bottles. These bottles were autoclaved, along with bottles containing 15 cm3 of
agar solution. The agar and salt solutions were kept in a water bath at approximately 50 oC
until ready to be used. The agar and salt solutions were mixed (ensuring the necks of the
bottles were flamed to burn any dust present). The agar/salt solutions were then poured
into sterile petri dishes, and the agar was left to set at room temperature. The petri dishes
were turned upside down to prevent condensation (which forms on the lid) dropping onto the
surface of the agar. The dishes were refrigerated until required.
Treatment of agar with bacteria using sterile technique:
All equipment was autoclaved before use. Three drops of bacterial broth containing E. coli
were placed in the middle of each agar surface using a sterile teat pipette. Before use, the
tip of the pipette was flamed. A sterile glass spreader was dipped in ethanol, and the
ethanol burned off immediately before the spreader was used. The broth was then spread
around the agar surface. Paper discs impregnated with antibiotic (streptomycin) were
dropped onto the surface of the agar using a dispenser. Four discs were placed in each petri
dish. Three plates for each concentration were treated with antibiotic discs, and one dish
for each concentration was treated with a control. The control consisted of discs of filter
paper (cut using a hole-punch). These were sterilised in an oven at 120 oC for an hour, and
were thereafter handled with sterilised forceps. After inoculation the petri dishes were
labelled, sealed, and incubated at 37 oC for twenty-four hours. After incubation, the
diameter of the circular areas around the discs which were free of bacterial growth were
measured using a pair of callipers.
Results:
Table 1 shows the diameter of each of the bacteria-free areas in each of the petri dishes.
The averages of these measurements are also shown.
Figure 2 shows the average diameter of the bacteria-free area against concentration of
sodium chloride in agar solution.

105
Table
1 Diameter
of bacteria-free
areas
when E.
coli was
incubated
with theOverall
antibiotic
Concentration
Streptomycin
Diameter
of bacteria
free-area
around
Average
Streptomycin
concentrations
sodium
solution,
twentyof salt in agar,in different
present
discs, (mm). of
Four
discs inchloride/agar
each petri
diameter
per foraverage
four
(molhours
dm-3) at 37 oC.
dish.
dish, (mm).
diameter,
[3 s.f.]
(mm).
0.0
No (control)
No growth inhibition
Yes
27
27
28
27.5
27.4
26.2
Yes
27
26
26
Disc fell
26.3
off
Yes
28
25
24
23
25.0
0.025
No (control)
Yes
26
27
26
26.5
26.4
24.8
Yes
25
25
25.5
25
25.1
Yes
24
25.5
25
17
22.9
0.0500
No (control)
Yes
27
25.5
27
28
26.9
25.4
Yes
26.5
26
26
25.5
26.0
Yes
24
23.5
23
23
23.4
0.0750
No (control)
Yes
28
27
27.5
26
27.1
25.9
Yes
26
26
25.5
Disc fell
25.8
off
Yes
24.5
25
25
25.5
25.0
0.100
No (control)
Yes
25.5
26
26
25
25.6
25.4
Yes
25
25.5
26
25.5
25.5
Yes
25
25
25.5
25
25.1
0.0125
No (control)
Yes
27
26.5
27
26
26.6
25.3
Yes
25
25
24.5
25
24.9
Yes
25
23
24.5
24.5
24.3
0.150
No (control)
Yes
27
27
26
25
26.3
25.7
Yes
27
27
26.5
26
26.6
Yes
24.5
24
24
24
24.1
0.175
No (control)
Yes
25.5
27
27
26.5
26.5
24.8
Yes
24
24
23.5
23.5
23.8
Yes
24
23
25
24
24.0
0.200
No (control)
0.225
0.250
0.375
106
Yes
No (control)
Yes
Yes
Yes
No (control)
Yes
Yes
Yes
No (control)
Yes
Yes
Yes
25
25
26
24.5
25
25
25
24.5
24.5
24
23
23
25
23
24
24
24
24.5
23
23
21
22
23
23
22
21.5
22
22
21.5
22
25
25.3
24.7
24.5
24.5
23
24.8
24.6
23.3
24.2
24.5
24.5
23
24.1
24.3
22.5
23.6
22
22
21.5
22.5
21.9
21.8
22.1

0.5
0.5
No
No (control)
(control)
Yes
Yes
Yes
Yes
Yes
Yes
No
No (control)
(control)
Yes
Yes
Yes
Yes
Yes
Yes
No
No (control)
(control)
Yes
Yes
Yes
Yes
Yes
Yes
0.625
0.625
0.75
0.75
27
No
No growth
growth inhibition
inhibition
21
21
22
21
21.3
21
21
22
21
21.3
21.3
21.3
22
21.5
21
21
21.4
22
21.5
21
21
21.4
20.5
21
21
22
21.1
20.5
21
21
22
21.1
No
No growth
growth inhibition
inhibition
No
No measurable
measurable growth.
growth. Light
Light smearing
smearing over
over plates.
plates. No
No visible
visible
inhibition.
inhibition.
No
No growth
growth inhibition
inhibition
23
21
23
21
21.5
21.5
21.5
21.5
21.5
21.5
21.5
21.5
22
22
21
21
21
21
22
22
21
21
21
21
22.0
22.0
21.3
21.3
21.3
21.3
21.5
21.5
Figure 2 Diameter of bacteria-free areas when E. coli was incubated

with the antibiotic streptomycin in different concentrations of sodium
o
chloride/agar solution, for twenty-four hours at 37 C
Diameter of area of bacterial growth

inhibition (mm)
26
25
24
23
22
21
20
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
-3
Concentration of sodium chloride in agar solution (mol dm )
Analysis
Figure 2 suggests a significant negative correlation between the concentration of salt in the
agar used, and the distance over which the antibiotic streptomycin is effective in preventing
growth of E. coli. The salt concentration is the only variable that was changed, and the
control plates (i.e. those with no antibiotic) showed growth of bacteria all over the surface
of the plates at every concentration. For example, the average diameter of the area of
growth inhibition with a salt concentration of 0.75 M was 21.5 mm, compared with 26.2 mm
for zero salt concentration. The data strongly suggest that salt concentration has a negative
effect on the action of the antibiotic used.
A linear regression line (R2 = 0.8312) was added to the plot of diameter of area of inhibition
against salt concentration. It shows a clear relationship; as the concentration of salt
increases, the area of inhibition decreases.
The Student t test can be applied to the means of two sets of data to show whether the
means are significantly different. The test was used to disprove the null hypothesis that

107
there is no significant difference between the means of the two sets of data for the
extremes of sodium chloride concentration.
t is defined by
(x1 x 2 )
t=
s12 s 22
+
n1 n 2
where s2 = variance
x = mean of diameter of area of inhibition
n = number of pieces of data
The variance (see Appendix D) of the diameters for 0.0 M salt is
s1 =
83232.25
11
(11 1)
7593.25
= 2.668
and for 0.75 M salt is
s2 =
2
66564
12
(12 1)
5551
= 0.3636
The value of t for the two extreme salt concentrations (i.e. 0.0 M and 0.75 M), is therefore:
t=
(26.2 21.5)
2.668 0.3636
+
11
12
= 9.00
For 21 degrees of freedom (the number of degrees of freedom is equal to the total number
of pieces of data used minus two (n1+ n2 2)), t = 9.00 is well above the value needed (2.080)
to ensure a confidence level of 5% in the rejection of the null hypothesis. (See Appendix D).
Similarly, there is a significant difference between the means of the diameters for the salt
concentrations of 0.1 M and 0.2 M. The value for t is 2.69 which is greater than the value
needed for a 5% confidence level at 22 degrees of freedom (2.074). Thus for each of these
pairs of data the null hypothesis is rejected. Increasing sodium chloride concentration
decreases the effectiveness of streptomycin against the growth of E. coli.
It is possible that the ions present in sodium chloride solution interfere with the functional
groups of the streptomycin molecule. Relatively electronegative ions such as chloride ions
may distort the shape of the streptomycin molecule by attracting electrons which are part
of the antibiotic molecule, thus changing the shape and/or properties of the streptomycin
molecule. Alternatively, it may be the sodium cation which causes the distortion. Some
divalent cations have been found to have a greater inhibiting effect on natural antimicrobial
proteins compared with monovalent cations.3 It has also been suggested that cations bind to
108

bacteria and so shield them from streptomycin molecules.3 It is plausible that these theories
also apply to medicinal antibiotics such as streptomycin. Streptomycin works by inhibiting
protein synthesis in the bacteria,4,5 which in turn prevents bacteria from growing. It is this
action of inhibiting protein synthesis which appears to be affected by the presence of salt
in the agar upon which the bacterial culture is growing. If this synthesis is no longer being
disrupted, then the bacterial growth will no longer be affected.
The effect of salt concentration on the action of antibiotics is relevant in a medical context.
The airway surface liquid (mucus) of patients with cystic fibrosis is thick, sticky and salty,
compared with the airway surface liquid of non-sufferers. The build up of this mucus can
lead to regular and persistent infections and CF sufferers are often prescribed antibiotics
to deal with these. The outcome of my investigation leads me to believe that the
effectiveness of antibiotics against bacterial infection may be reduced in CF patients
because the antibiotic is required to work in saltier conditions than normal. In people with
abnormal cystic fibrosis transmembrane regulatory (CFTR) proteins, chloride ions pumped
into epithelial cells of the airway cannot leave. The chloride concentration in the cell
increases, and consequently so does the sodium ion concentration, due to the attracting
electrical charges on these ions. The increased sodium chloride concentration in the cell
causes water from the mucus lining the airway to move into the cells due to osmosis. This
movement of water from the mucus lining the airway causes the mucus to be more
concentrated in sodium chloride than would be the case if the CFTR protein was normal.6
Evaluation:
The investigation was successful in disproving the null hypothesis to a 5% level of
confidence, but the results obtained do have limitations. The range of concentrations of salt
used was wide enough to show a trend. However, in order to be more confident of the
conclusion, I believe a greater range of concentrations would be needed. The graph of
diameter of the area of inhibition against salt concentration shows a certain degree of
scatter around the linear regression line. In order to minimise this scatter, more repeats of
each measurement should be carried out. This would increase the reliability of the results.
The outcome of the Student t test would be more reliable if there were more than twelve
data points for each salt concentration. If the investigation were to be repeated, more
experiments would be carried out between 0.375 M and 0.75 M salt concentrations. Fewer
different concentrations were made between these values because of time limitations on the
number of experiments that could be carried out.
The practical technique used was designed to ensure a fair test. The distribution of salt in
agar solution on each plate was homogenous, as the salt solutions were mixed thoroughly with
the agar before the plates were poured. The petri dishes were taken from a sterile pack.
Callipers were used to measure the diameters of the areas of inhibition, which is more
accurate than using a ruler. However, because the callipers can only measure to within 0.5
mm, a travelling microscope could be used instead in a repeat of the investigation to allow a
greater degree of accuracy. The area of inhibition was assumed to be circular, and the
density of growth to be uniform. An improvement to the method could be to measure the
area covered by bacterial growth using a light intensity technique. This would allow slight
growth in the area of inhibition to be taken into account.
A possible explanation for the relatively poor growth of bacteria all over the plates at 0.625
M salt concentration is that the bacterial broth was not entirely homogenous, hence drops
of bacterial broth used to inoculate some plates were much less concentrated than others,
resulting in differences in the growth of bacteria.

109
Word count: 1997

REFERENCES:
1. http://alwaysyourchoice.com/ayc/childrens/children/fibrosis.php (viewed on 8/4/04).

2. http://www.uihealthcare.com/news/news/2000/10/1016cfxylitol.html (viewed on
8/4/04).
3. Travis, S. M., Conway, B.-A. D., Zabner, J., Smith, J. J., Anderson, N. N., Singh, P. K.,
Greenberg, E. P. and Welsh, M. J., Activity of Abundant Antimicrobials of the Human
Airway, American Journal of Respiratory Cell and Molecular Biology, 20 (1999) 872-879.
4. Bauman, R. W., Microbiology, Pearson Education, 2004
5. Murray, P. R., Rosenthal, K. S., Kobayashi, G. S. and Pfaller, M. A., Medical Microbiology,
4th ed., Mosby, 2002.
6. Salters-Nuffield, Advanced Biology AS Student Book 1, pilot edition, Heinemann
Educational Publishers, 2002.
APPENDIX A.
[Risk assessment supplied by student]
APPENDIX B.
Preliminary experiments:
A preliminary experiment was carried out to determine which bacterial culture to use.
Escherichia coli and Staphylococcus albus were initially both grown with discs of penicillin as
the antibiotic. Sufficient growth of S. albus bacteria did not occur, even in the control
plates (i.e. those with 0 M concentration of sodium chloride). Growth of E. coli was greater,
so I therefore decided to use Escherichia coli in the investigation in order to obtain
measurable results. The antibiotic discs did not seem to have a substantial effect on the
growth of the bacteria. E. coli is a gram-negative organism, and so it is relatively resistant to
penicillin, due to the absence of a thick peptidoglycan layer in the cell wall 5. As penicillin
inhibits bacterial growth by disrupting the synthesis of cell walls, a different antibiotic was
needed to act against the bacteria. Streptomycin inhibits protein synthesis, and is
therefore effective against gram-negative bacteria such as E. coli 4. Because there was
relatively little bacterial growth after several days, I decided to use three drops of
bacterial broth per plate, rather than only one. Another advantage of doing preliminary work
is to gain experience in practising sterile technique.
APPENDIX C.
Example calculation for mass of sodium chloride to be dissolved to make standard solutions.
Where n = number of moles of NaCl
c = concentration of solution (mol dm3)
v = volume of solution (dm-3)
m = mass of NaCl (g)
M = molar mass of NaCl
110

For 100 cm3 of 0.1 M solution:
n = cv
n = 0.1100 = 0.01 moles

1000
m = nM
m = 0.01 (23 + 35.5) = 0.585 g

Therefore 0.585 g of sodium chloride was dissolved in 100 cm3 to form a solution of
concentration
0.1 mol dm-3.
Mass of sodium chloride added to 100 cm
Concentration
of solution
before adding
to agar
(mol dm-3)
Mass of
sodium
chloride
dissolved (g)
of water
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
0.9
1.0
0.585
1.17
1.76
2.34
2.93
3.51
4.10
4.68
5.27
5.85
Concentration
of solution
before adding
to agar
(mol dm-3)
1.5
2.0
2.5
3.0
Mass of
sodium
chloride
dissolved (g)
8.78
11.7
14.6
17.6
APPENDIX D.
The variance of a population is found using the following formula:
( x )
s =
2
(n 1)
where s2 = variance
s = standard deviation
x = dependant variable (diameter of area of inhibition)
n = number of pieces of data

111
The values of t needed for each confidence level at the appropriate number of degrees
of freedom.
Number of
degrees of
freedom
21
22
20%
10%
Confidence level
5%
2%
1%
0.1%
1.323
1.321
1.721
1.717
2.080
2.074
3.819
3.792
2.518
2.508
2.831
2.819
Total 6/6
Moderators comments
112
This exemplar achieved maximum marks.

Student checklist for project skills (Unit 6135/02)

While doing your project and before finally submitting your coursework you may find
it useful to check the points listed below.
Preliminary work
Have you:
read your teachers comments on your AS coursework
looked at the Edexcel website
read the coursework criteria
thought of a title/aims?
Research and rationale

Have you:
consulted several sources
consulted websites, noted their addresses and the date you accessed them
used books as well as the internet and recorded details of the books used
discussed your ideas with your teacher
selected material carefully
said how you have used the information you have gathered in your planning
stated your rationale clearly
stated the aims of the experiment clearly
derived or explained any relevant theory
checked that the theory is correct and at full A-level standard?
Planning
Have you:
listed the apparatus you will use
drawn and labelled a diagram
considered safety
stated the variables which could affect your experiment
explained which variables will vary and which will be kept constant
explained what you will measure
said what instruments/apparatus you will use and why
described clearly any apparatus you have devised
considered the sensitivity of the measuring instruments

113
explained the accuracy you will need to measure to
described how you will treat your results
carried out a pilot experiment
reviewed your plan in view of the pilot
selected a suitable statistical test to apply to the results of your experiment?
Implementing
Have you set up the apparatus correctly?
Is your working area well organised?
Have you tables ready drawn for your results?
Are you working with care?
Are you being methodical?
Are you looking for trends as you take results?
Have you reviewed your planned procedure and made any required modifications?
Observing and recording

Have you:
included units
taken at least five sets of measurements
checked for anomalous readings
repeated measurements if appropriate
recorded measurements to an appropriate number of significant figures
limited the number of significant figures, if you used a spreadsheet for your
results
been consistent in the number of significant figures used
explained any changes to your planned procedures?
Interpreting and evaluating
Have you drawn at least one graph?
Is it at least half an A4 sheet in size?
Is the graph a straight line or a curve?
If you used a computer to draw the graph, does the graph have a grid and a
sensible scale?
Have you labelled your graph and included units?
Have you put multiple graphs on to the same axes to help comparisons?
Have you identified trends and patterns?
Have you carried out a suitable statistical test?
Have you explained the biology involved (you may have done this in your plan)?
114

Have you stated a conclusion?
Have you linked your conclusion to your results and to your aims?
Did you include units in any calculated values?
Have you discussed limitations and conclusions qualitatively (words) or

quantitatively (numbers)?
Has any calculated value been stated to a sensible number of significant figures?
Have you numbered the pages?
Communicating
You may find it useful to add an abstract (summary) along with a contents section at
the start.
Is your report word processed?
Can you submit it on disc or electronically?
Is the structure of your report clear?
Have you used subheadings?
Are the aims and conclusions clear?
Have you included labelled graphs, tables and diagrams?
Is there a bibliography?
Have you numbered the pages?
Have you proofread the report?
Is your report no longer than 2000 words?
1024rl150206:TPD\SUPPORT\UA016953 GCE IN BIOLOGY (SALTERS-NUFFIELD) CG.DOC.1-42/4

115
Further copies of this publication are available from

Edexcel Publications, Adamsway, Mansfield, Notts, NG18 4FN
Telephone 01623 467467
Fax 01623 450481
Email: publications@linneydirect.com
Publications Code UA016953 February 2006
For more information on Edexcel and BTEC qualifications please contact
Customer Services on 0870 240 9800
or http://enquiries.edexcel.org.uk
or visit our website: www.edexcel.org.uk
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Registered Office: One90 High Holborn, London WC1V 7BH

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GCE

Edexcel Advanced Subsidiary GCE in

Edexcel Advanced GCE in

Biology (Salters-Nuffield) (8048/9048)

Edexcel Advanced Subsidiary/Advanced GCE

Authorised by Jim Dobson

Visit or issue report (Unit 6133/01)

Identifying and describing two aspects of biology

Account of biological principles

Student checklist (Unit 6133/01)

Practical work review paper (Unit 6133/02)

Safety and use of apparatus

Producing reliable and valid results

Using results to draw conclusions

Choosing the best practical

Coursework investigation (Unit 6135/02)

Assessment criteria Research and rationale

Exemplar 1 Jennys work

Exemplar 2 Michaels work

Assessment criteria Planning

Exemplar 3 Samirs work

Exemplar 4 Robyns work

Assessment criteria Implementing

Exemplar 5 Marcos work

Exemplar 6 Shazias work

Assessment criteria Observing and recording

Exemplar 7 Susans work

Exemplar 8 Gabriels work

Assessment criteria Interpreting and evaluating

Exemplar 9 Jatinders work

Exemplar 10 Davids work

Assessment criteria Communicating

Exemplar 11 Rosannas work

Exemplar 12 Jans work

Exemplar 13 (maximum mark) Katharinas work

Student checklist for project skills (Unit 6135/02)

UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

Visit or issue report (Unit 6133/01)

UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

Identifying and describing two aspects of biology

The purpose or significance of two aspects of biology are described

There is some appreciation of the ethical, social, economic or

The significance of both aspects of biology are described clearly

There is a clear explanation of the ethical, social, economic or

Exemplar A A visit to Batemans Brewery

UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

the importance of glycolysis, with diagrams and details of the biochemistry

the importance of temperature for fermentation, ie warmer temperatures mean

specific gravity changes

secondary fermentation and other uses of fermentation.

UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

In order to gain higher marks the student would need to:

UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

Exemplar C Deforestation: the global assault continues

In order to gain higher marks the student would need to:

Exemplar D Information leaflet on glaucoma

UA016953 Coursework guide Edexcel Advanced Subsidiary/Advanced GCE in Biology (Salters-Nuffield)

Student D clearly identifies the two aspects of biology under investigation.

In order to gain a higher mark the student would need to:

mention that untreated diseases, such as glaucoma, could result in premature

Exemplar E Biology issue report

There is nothing on the economics of the treatments.

In order to gain higher marks the student would need to: