Impact of Light Level on Biodiversity of Leaf Litter

Rationale
Biodiversity encompasses the variety of living organisms and ecosystems on Earth, which provide essential services
such as pollination, nutrient cycling, and water purification (Tilman, Isbell, & Cowles, 2014). Measuring biodiversity
is crucial to understand the health and functioning of an ecosystem, and can be done in many different ways, including
species richness, evenness, and Simpson's Diversity Index (SDI) (Magurran, 2013). SDI takes into account the number
of species present and their abundance and ranges from 0 to 1, with higher values indicating greater diversity. SDI is
calculated using the formula:
∑ 𝑛(𝑛 − 1)
𝑆𝐷𝐼 = 1 −
𝑁(𝑁 − 1)
where n is the number of individuals of a particular species and N is the total number of individuals across all species.
There are multifarious factors that can influence biodiversity, both biotic and abiotic in nature; climate, habitat
fragmentation, and pollution are some of the most prominent. Canopy type, owing to its ability to modify exposure to
light and heat, can influence the diversity of species in leaf litter (Stork, 1993). The presence of perennial streams in
forested areas can have a significant impact on local biodiversity, as they provide a stable source of water and support
a range of aquatic and terrestrial species (Boulton, et al., 2008). Leaf litter is an essential component of forest
ecosystems, made up of a cornucopia of dead leaves, twigs, and other organic material that accumulate on the forest
floor. Leaf litter, with its inherent diversity, provides habitat and serves as a source of sustenance for a plethora of
invertebrates (Sánchez‐Galindo, et al., 2021).
The Tullgren Funnel method is a popular technique for extracting invertebrates from leaf litter. Heat is applied to the
litter in a funnel, causing the invertebrates to fall through and into a container with a preserving liquid (Krebs, 2019).
In open canopy forests, the SDI is anticipated to ascend when lux values exceed 250, owing to the heightened
reception of sunlight. This increased luminosity has the potential to stimulate the proliferation of disparate flora
species, thereby endowing invertebrates with multifarious habitats. Conversely, closed canopy forests are expected to
exhibit a diminished SDI when lux values dip below 250, due to the concomitant attenuation of solar exposure. The
constricting light availability could lead to a reduction in the diversity of plant species, with invertebrates being
subjected to less diverse ecological niches consequently.
Research Question
To what extent does the canopy type impact the Simpson's Diversity Index (SDI) of the species found in a bucket of
leaf litter gathered from a forest that encompasses a perennial stream, when the canopy type is bifurcated based on
Lux values exceeding (open canopy) or falling below (closed canopy) 250?
Original Experiment
In the original experiment, a sample of leaf litter was collected from a 1m2 quadrat from both a natural and artificial
location to determine whether the extent of human interference affects the species richness. It was found that the
artificial site had lower species richness, whilst the natural site had higher species richness. However, species richness
is only a moderate indicator of biodiversity as it only measures one facet of biodiversity, therefore this indicator was
changed to SDI in the modified experiment. Furthermore, the dependant variable was changed to canopy type, as this
could be quantified using Lux whilst human interference was a qualitative variable.
Modifications

Modification
Increased number of trials from one to five
Poured insects on paper before counting as opposed to
counting them in a beaker.
Increased amount of time light was on from one hour to
one day
Justification
Improves reliability of data as it reduces the impact of
outliers and random error on the mean.
Improves accuracy of counting measure which improves
reliability and validity of results.
Ensures all insects have enough time to fall out of the
leaf litter, which reduces effect of error on the mean,
improving reliability.

Safety

Risk Prevention Management

Cuts from broken or chipped
glassware.
Potential skin, breathing, or eye
irritation from inhaling or coming
into contact with Methylated spirits
-Handle glassware and beakers with
care and caution.
-Keep glassware away from the edge
of lab benches.
-Have a sharps bin nearby and
dispose of any broken glass
immediately.
-Inspect and discard any chipped or
cracked beakers.
-Do not put organic solvents in
plastic containers.
-Handle the methylated spirits with
care when transporting it.
-Ensure methylated spirits is only
handled for 10–20-minute periods.
-Have a working eyewash nearby.
-Alert the supervisor.
-Apply the appropriate first aid and
seek additional medical attention if
required.
-Alert supervisor.
-Rinse off the area.
-Remove fabric drenched in
methylated spirits.
-Use eyewash.
-Seek extra medical attention if
required.

Ethical Considerations
It was determined necessary to kill insects for the experiment to make conclusions. To minimize the pain caused to the
insects, concentrated methylated spirits were used. This method was selected because it increased the speed of death,
making the experiment more humane.
Sample Calculations

Calculation Example
Simpson’s Diversity Index ∑ 𝑛(𝑛 − 1)
𝑆𝐷𝐼 = 1 −
𝑁(𝑁 − 1)
48
=1−
90
= 1 − 0.533
= 4.667
Average SDI (0.467 + 0.750 + 0.222 + 0.667 + 0.733)
𝑆𝐷𝐼𝑚𝑒𝑎𝑛 =
5
(2.839)
=
5
= 0.568
P Value The P value was calculated in excel by using the
“t-Test: Two-Sample Assuming Unequal Variances”
function and the five SDI values for both canopy types.

𝑃 = 0.306
SDI Uncertainty 1 − 𝑆𝐷𝐼
𝑢𝑛𝑐𝑒𝑟𝑡𝑎𝑖𝑛𝑡𝑦 =
√(𝑁 − 1)
1 − 0.467
=
√(10 − 1)
0.533
= 3
𝑢𝑛𝑐𝑒𝑟𝑡𝑎𝑖𝑛𝑡𝑦 = 0.17778
Processed Data

Lux (±0.05) Total Organisms SDI Average

7047 10 0.46667 ±0.17778
935 8 0.75000 ±0.09449
543 9 0.22222 ±0.27499
543 3 0.66667 ±0.23570
367 6 0.73333 ±0.11926 0.567777778
111 99 0.57205 ±0.04323
111 12 0.54545 ±0.13705
87 45 0.69798 ±0.04553
39 47 0.42461 ±0.08484
22 61 0.27158 ±0.09404 0.502334578

Trends, Patterns, Relationships

0.9

0.8

0.7

0.6
SDI

0.5

0.4

0.3

0.2

0.1

0
Open Canopy Closed Canopy
Canopy Type

Figure 1: Average SDI vs. Canopy Type Scatterplot

Interpretation: The relationship between SDI and Canopy Type was expected to show a significant difference in
biodiversity between the two canopy types. However, the data does not fit this trend, as the average SDI for open
canopy is only slightly higher than for closed canopy, but not significantly different. This is supported by the P value
of 0.306, which is above the threshold of 0.05 for statistical significance. Further experimentation and data analysis
may be necessary to confirm any potential relationship between canopy type and SDI.
Analysis: The bar graph comparing SDI values for open and closed canopies shows a negligible difference between
the two. The P value, which measures the probability of obtaining the observed results if there is no real difference
between the two groups, was 0.306. Any P value above 0.05 is considered not statistically significant, indicating that
there is no significant difference in SDI values between the two canopies. Additionally, the error bars for both groups
are quite large and overlap extensively, which raises concerns about the reliability of the data.

Sources of Error
Effecting reliability:
The mass, depth, and age of the leaf litter samples collected were not consistent, which could have affected the results
of the study and thus biasing the data.
Some samples of leaf litter were collected near ant nests, pathways, or areas that were more easily accessible, which
could have influenced the types of organisms collected. Additionally, the individuals who collected the leaf litter and
counted the organisms were not trained, which could have introduced human error. Finally, organisms crawled out of
the buckets when being transported or placed under a heat lamp, potentially resulting in the loss of organisms.

Effecting validity:
The lack of correlation between SDI and canopy type expected values are significant limitations to the validity of the
experimental process. The unreliable data and lack of correlation could have arisen from factors such as inconsistent
lighting conditions or inaccurate measurements, which could have influenced the results, leading to inaccurate
conclusions. Consequently, the validity of the data obtained from the experiment may be questioned.
The use of leaf litter as a measure of biodiversity and the selection of a 1m2 quadrat as the optimal area for sampling is
also a limitation. The selection of a single area to represent the biodiversity of the entire forest may not provide an
accurate representation of the overall biodiversity of the ecosystem. The use of leaf litter as a measure of biodiversity
is also limited, as it only accounts for the organisms living on or near the surface and may not account for those living
deeper within the soil. This limitation may lead to inaccurate conclusions regarding the overall biodiversity of the
ecosystem.
Suggested Refinements and Extensions
Refinements:
Ensure uniformity in leaf litter mass, depth, and age by collecting samples from the same location at the same time of
year and weighing and measuring the litter before placing it under the heat lamp. This will decrease variation in the
amount and composition of litter, which could have affected the results.
Standardize the leaf litter collection process by training all collectors to follow the same protocol and using the same
people for all collections. This will decrease the chance of external factors such as ant nests, pathways, or individual
collection techniques affecting the results.
Use a data logger to measure and record the actual lux levels at each quadrat over a longer period of time. This will
allow for a more accurate understanding of the lux levels in the forest and may reveal any trends or patterns in the
variation.
Extensions:
Conduct a comparative analysis of different sampling methods for measuring biodiversity in the same forest
ecosystem. For example, compare the results of the leaf litter method used in this experiment with results obtained
from pitfall traps or sweep nets. This will provide a better understanding of the strengths and weaknesses of each
sampling method, and which one may be more appropriate for different research questions or study sites. Additionally,
it will allow for a more robust assessment of the biodiversity in the forest ecosystem.
Expand the quadrat size to 2m2 or 4m2 and compare the results to those from the 1m2 quadrats. This will allow for a
better assessment of whether the 1m2 quadrats were the optimal size for this experiment and increase the
generalizability of the results to other forests.
Conclusion
In conclusion, the evidence suggests that there is very little correlation between SDI and canopy cover. This is
contrary to the literature suggesting that there should be an increase in SDI as canopy cover increases. The scatterplot
of SDI vs. Lux showed no trend and random points, with an R2 value of 0.0382. The bar graph of SDI vs. canopy type
showed a small difference between open and closed canopy, but the P value of 0.306 suggests that this difference is
not significant. The large error bars and overlapping standard error values in the processed data table also suggest low
precision and reliability of the findings. The experiment was flawed, and further analysis should be explored to better
understand the relationship between canopy type and SDI in the forest surrounding the perennial stream.

References
Boulton, A. J., Boyero, L., Covich, A. P., Dobson, M., Lake, S., & Pearson, R. (2008). Tropical Stream Ecology.
Inland Waters.
Convention on Biological Diversity. (2021, January 12). What is Biodiversity? Retrieved from Convention on
Biological Diversity: https://www.cbd.int/biodiversity/
Krebs, C. J. (2019). Ecology: The Experimental Analysis of Distribution and Abundance: Pearson New International
Edition. Pearson.
Magurran, A. E. (2013). Measuring Biological Diversity. John Wiley & Sons Ltd.
Sánchez‐Galindo, L. M., Sandmann, D., Marian, F., Krashevska, V., Maraun, M., & Scheu, S. (2021, January 29). Leaf
litter identity rather than diversity shapes microbial functions and microarthropod abundance in tropical
montane rainforests. Retrieved from Ecology and Evolution:
https://onlinelibrary.wiley.com/doi/10.1002/ece3.7208
Stork, N. E. (1993). How many species are there? Retrieved from Biodiversity and Conservation.
Tilman, D., Isbell, F., & Cowles, J. M. (2014). Biodiversity and Ecosystem Functioning. Retrieved from Annual
Review of Ecology, Evolution, and Systematics: https://www.annualreviews.org/doi/abs/10.1146/annurev-
ecolsys-120213-091917
Appendix
Appendix 1: Raw Data
Lux Bug
Amphipod Centipede Slater Mite Earwig Beetle Larva Beetle Ant Slug Millipede Symphylan White Worm Flatworm Heteroaryl Cockroach Springtail Total

7047 0 0 3 0 0 0 0 7 0 0 0 0 0 0 0 0 10
935 0 0 0 0 0 1 1 4 0 0 0 0 2 0 0 0 8
543 0 0 0 0 0 0 1 8 0 0 0 0 0 0 0 0 9
543 0 0 0 0 0 0 0 1 0 0 0 0 2 0 0 0 3
367 0 0 2 0 0 0 0 3 0 0 0 0 1 0 0 0 6
111 46 0 4 0 0 1 0 0 0 0 1 0 1 0 0 46 99
111 8 0 2 0 2 0 0 0 0 0 0 0 0 0 0 0 12
87 17 0 3 0 2 0 0 4 0 0 0 0 0 0 1 18 45
39 35 0 0 0 0 0 0 7 0 1 0 0 0 0 0 4 47
22 2 0 0 0 1 0 0 52 0 0 0 1 4 1 0 0 61