On-line Practical: Acorn barnacle (Semibalanus balanoides) and the

effects of microplastics on feeding rate.

Submission date: Upload to Turnitin by: Week 6, 1630 11th March
Answer the questions, have the completed table and bar graph in a
new word document.

Objective of this Practical Assessment is to a) develop your ability

to present data and b) critically assess how an experiment has been
conducted.
Background information:
Barnacles are important crustaceans that can be found in shallow-waters, down to the
ocean depths. In coastal waters they can be found in very high densities attached to rocks.
They are suspension-feeders removing plankton and detritus from the water-column.
However, large quantities of fine plastics are now present in coastal waters as well as their
natural prey.
With my lock-down boredom, I decided to conduct an experiment looking at cirral beating
rate in barnacles in relation to plastics exposure. I used digital-video camera equipment and
conducted this experiment in my ‘home-lab’ with limited other equipment. Half-way
through, I realised there was a boxset I had not watched on Amazon yet and gave up!!
I have left you to analyse the rest of the videos, complete the data table & answer some
questions! The full details of the Materials and Methods used, and Procedure followed is
detailed below.

Dr Cowie’s, home microplastics experiment procedure

Materials and Methods:
1. Species: Semibalanus balanoides; collected from the intertidal zone of Lunderston Bay,
Inverkip
2. Barnacles were on small stones, all different sizes of barnacle.
3. Natural seawater was collected at the same time, salinity 32.
4. Experiments conducted at water-temperatures between 10 – 15 0C
5. Grated polyvinyl chloride lids, particle-size of dust = 150-400 µm.
6. Forceps, pipettes
7. Digital, HD video-recording microscope (up to x200 magnification)
8. ‘Clicker-counter’ free app for counting the number of beats.
Procedure:
1. A small stone with barnacles on it, was placed in one of the small cylindrical
containers.
2. Seawater (100C – 150C) was then poured into the container so that it just covered the
barnacles allowing them to feed.
3. The barnacles were given 5 minutes to settle down, then recording was started.
4. 10 close-up recordings of different, randomly selected, feeding barnacles were
taken.
5. After the 10 control recordings were taken, the barnacles were exposed to
microplastics.
6. A ‘pinch’ of plastic polyvinyl chloride dust (150 -400µm), was added to the water
surrounding the barnacles.
7. 10 more, close-up recordings of barnacles feeding in the water seeded with plastics
were taken.
8. 14 of these videos were analysed by recording the number of cirral beats over the
two minutes and average no. of beats per minute. Was then determined. Counts
were made using a free clicker-counter app.
9. The average beats per minute were then inserted into a table.

Microplastics Exercise:
Resources available to you: This assessment guidance document
1. Workshop 3 power-point slides.
2. Six, 2-minute video clips (3 x Control, 3 and Experimental)
3. Website link to credible source with information about barnacles:
https://www.marlin.ac.uk/species/detail/1376
4. Research paper given to you previously.
5. The week 2 lecture section on the impacts of plastics in the environment.
 Practical assessment elements:
Where possible support your answers with referenced information.
Element 1: (20 marks total)
a) You need to watch the remaining 6 videos. Watch each video, and record the number of
times that the selected barnacle beats over the two minutes of the video. Then divide
this value by two to give the average number of beats over 1 minute e.g. beats per
minute-1.
b) Fill in the partially completed table below, Table 1 (when marking I will have a ± 2%
tolerance for no. of beats).

Table 1: Partial table of raw data, from recordings of barnacle (Semibalanus

balanoides) beats minute-1. Control recordings (no plastics) and Experimental
recordings (plastics introduced).

Replicate number Control (bpm) Experimental (bpm)

1. 90 51
2. 75 49
3. 65 38
4. 69 53
5. 23 77
6. 75 34
7. 52 45
8. Video C2: 5 Video E2: 78
9. Video C3: 28 Video E3: 14
10. Video C1: 10 Video E1: 56
Mean value 49.20 49.50
95% confidence 17.83 11.22
intervals

Element 2: (20 marks total)

a) Present the mean values and 95% confidence intervals from the table in a vertical
bar chart. N.B. Look at slide 19 in the Week 3 PowerPoint to see the elements that
should be included.
Figure 1. The bar chart of the mean of beats per minute-1 (+/- 95% CL). There is no significant
difference between the controlled and experimental beats as the confidence limits overlap.
Error bars are standard deviation. N=10.

Element 3: (15 marks total)

a) Examine your graph, referring to the mean values and 95% confidence intervals,
explain what it shows about the response of the barnacles to exposure to
microplastics in the water (10 marks).
95 % Confidence intervals were used to check a statistical difference of beating rates
between barnacles that were not exposed to microplastics and barnacles that were exposed
to microplastics. The plotted bar chart of the mean of beats per minute -1 (+/- 95% CL)
showed that there is no significant difference between the controlled and experimental
beating rates as the confidence limits overlap. The mean values between the controlled and
experimental beating rates show no response of the barnacles to exposure to microplastics
in the water. The beating rate of individual barnacles may affect the results.
b) How has the behaviour of individual barnacles effected the results? (5 marks).
The beating rates of randomly selected, feeding barnacles vary between individual
barnacles. I would perform a different statistical test to draw a valid conclusion of statistical
significance. Each barnacle may have a different response of beating rates between
barnacles that were not exposed to microplastics and barnacles that were exposed to
microplastics.

Element 4: (10 marks total)

a) What statistical test could you use to establish whether there is a significant difference
between the mean number of beats displayed by control barnacles and experimental
barnacles (5%).
I would use the T-test to establish a significant difference between the mean of beating
rates between controlled and experimental barnacles. This statistical test would compare
the mean and standard deviation to find out a significant difference between two samples.
This would enable to see whether the differences between the controlled and experimental
barnacles are most likely not due to chance or sampling error.
b) Explain why you chose this test (5%).
I chose the T-test because it enables to produce a T-value which shows how many standard
units are separated by the means of the two groups. The higher the T-value, the more likely
the two samples are to be significantly different from each other. Calculation of T-value
enables to accept or reject the null hypothesis.

Element 5: (15 marks total)

In one of the videos, the introduced plastics are having a direct effect on the behaviour of
the shallow-feeding barnacle’s behaviour. Answer the questions below:
a) Describe, using scientific terms, what happens when the barnacle encounters floating
microplastics (5 marks)
When the barnacle encounters floating microplastics, a reduced uptake of oxygen can be
identified. The barnacle may take up microplastics while respiratory pumping which may be
the cause of limited oxygen uptake.
b) What could this behavioural response mean for overall future barnacle populations in the
seas? (5 marks)
Barnacle populations can be affected negatively by the long-term consumption of
microplastics. Encountering floating microplastics by barnacles cause a reduced uptake of
oxygen which may cause death when consumed for some long period of time. Therefore,
long term consumption of microplastics may reduce the number of barnacles in the seas
causing them to become endangered species.
c) What could the consequences of microplastic consumption be to barnacles? (5 marks)
The consequences of microplastic consumption to barnacles may include negative effects of
plastic ingestion such as limited oxygen uptake or intestinal blockage. Ingestion of
microplastics could also lead to an increase toxic exposure caused by high levels of toxins
contained by microplastics.

Element 6: (20 marks total)

a) Critically assess the materials, methods and procedures used to carry out the experiment.
Pay particular attention to the number of replicates used, environmental conditions, and
suggested improvements, use 3 referenced sources to support the points you are making.
N.B. Think also about the videos and the behaviour of individual barnacles when you are
talking about replicates.
The aim of the experiment was to see the effect of cirri beating rates in Semibalanus
balanoides. The number of beating rates were recorded between the barnacles that were
not exposed to microplastics and barnacles that were exposed to microplastics. The
barnacles feed by extending cirri (thoracic appendages) out from their shell for filtering
zooplankton and some similar sized particles from the water (White, 2008). The behaviour
of barnacles feeding changes over different flow rates (Tillin, 2015). However, the
experiment was conducted using still water only. In the absence of current the barnacle
creates its own by rhythmically beating the cirri to cause the water to move (Jones, 2011).
Flow tank with different velocities could also be used to see the effect of feeding rates on
barnacles. Barnacle feeding in the flow tank would increase with increasing flow speed.
Therefore, flow rate is an important factor which may affect feeding rates (Tillin, 2015).
Barnacle feeding activity is strongly dependent on environmental conditions. Food
concentration is another important factor which may affect feeding rates. Barnacle feeding
increases with increased food concentration. Therefore, qualitative and quantitative
differences in food supply also influences feeding activity. In the actual experiment only one
type of water was used. Barnacle feeding may vary across high and low food concentrations
and therefore, different types of water should be used to improve the experiment. (Sanford
et al., 1994). Furthermore, the mean values between the controlled and experimental
beating rates showed no response of the barnacles to exposure to microplastics in the
water. The number of replicates used was too small as the beating rate of individual
barnacles could affect the results. Therefore, the sample size should be increased to
improve the conducted experiment. Increased sample size can estimate the population
mean with more precision than with smaller samples (Mc Leod, 2019).
References
Jones, C, (2011). 'Acorn Barnacle (Semibalanus balanoides)', Ireland’s Wildlife, (May).
Available at: https://irelandswildlife.com/acorn-barnacle-semibalanus-balanoides/
(Accessed: 10 March 2021).
McLeod, L. (2019). 'What are Confidence Intervals in Statistics?' Simply Psychology, (June).
Available at: https://www.simplypsychology.org/confidence-interval.html (Accessed: 10
March 2021).
Sanford, E., Bermudez, D., and Gaines, S. (1994) 'Flow, food supply and acorn barnacle
population dynamics', Marine Ecology-progress Series, 104(1-2), pp. 49-62. DOI:
10.3354/meps104049
Tillin, H.M. (2015). Semibalanus balanoides and Littorina spp. on exposed to moderately
exposed eulittoral boulders and cobbles. In Tyler-Walters H. and Hiscock K. (eds) Marine Life
Information Network: Biology and Sensitivity Key Information Reviews, [on-line]. Plymouth:
Marine Biological Association of the United Kingdom. DOI:
https://dx.doi.org/10.17031/marlinhab.1026.1
White, N. (2008). Semibalanus balanoides. An acorn barnacle. In Tyler-Walters H. and
Hiscock K. (eds) Marine Life Information Network: Biology and Sensitivity Key Information
Reviews, [on-line]. Plymouth: Marine Biological Association of the United Kingdom.
(Accessed 10 March 2021). Available at: https://www.marlin.ac.uk/species/detail/1376