Effect of Flow Speed and Leaf Type on

Decomposition of Sweetgum and

Sycamore Leaf Packs
BIOL 271L-02
Kaley Christman
Kylie Wells, Cicely Clark, Sophie Panagakos

St. Mary’s College of MD

04/26/2023
Abstract
Leaf decomposition plays an important role in a stream’s ecosystem, overall health, and
nutrient levels. Leaves in streams often end up in clumps called leaf packs. Studying the
composition of these packs by creating some artificially in a laboratory setting can provide
insights into the stream and the leaves themselves. In this study, we tested American sweetgum
and American sycamore leaves in fast riffle and slow pool sections of St. Johns Creek to see if
the water flow rate or the type of the leaf had a greater impact on overall leaf decomposition. We
hypothesized that the leaves in the riffle regions will show more decomposition than those in the
pool regions due to the faster-moving water aiding decomposition and macroinvertebrates, and
we also hypothesized that the sycamore leaf packs will lose more mass than the sweetgum leaf
packs due to the higher surface area of the sycamore leaves and the higher nutrient content. To
test this, our study included 60 artificial leaf packs, 30 of each leaf type. We placed half of each
leaf type in a fast riffle section of the stream, and the other half in a slow pool. Every other week
for 8 weeks, leaf packs were collected and processed in order to find the ash-free dry weight of
each sample. Using this data, a linear regression and four T-tests were performed. Based on these
results, the null hypothesis was rejected, and our first hypothesis stating that leaves in the riffle
regions will show more decomposition than those in the pool regions was accepted, however our
second hypothesis that the sycamore leaf packs will lose more mass than the sweetgum leaf
packs was not accepted. Our conclusions can be applied to future studies about nutrients and
eutrophication and can show how stagnant pools in streams affect nutrient levels compared to
faster riffle sections.

Introduction

Leaf packs occur naturally when leaves enter streams and build up between things such
as rocks or branches within the water (Department of Biology, 2023). A build up of leaves into
these leaf packs can have very negative effects on the health of a stream. Increased levels of
excess nutrients, such as nitrogen and phosphorus, leads to eutrophication which reduces the
level of oxygen in the body of water. This has a negative impact on the organisms living in the
stream, as well as those that drink the water (Thiele 2018). However, these bunches of leaves
create an ideal environment for macroinvertebrates such as shredders, which as their name
suggests, shred the leaves as they feed. One of the major roles of shredders is that as they feed,
they break down coarse particulate organic matter, CPOM, into smaller bits known as fine
particulate organic matter, FPOM (Cummins et. al, 1989). This leads to faster decomposition of
the leaves. Additionally, different parts of the stream's water flow can have different effects on
leaf decomposition. These leaves in lower-order streams are subject to degradation factors such
as physical and microbial degradation, as well as invertebrate fragmentation (Graça 2001).
The degradation of leaves has been studied in many different contexts in recent years, with many
noting the important connection between the leaves and factors in the environment such as water
speed and depth, temperature, and more. In our study, we are comparing two types of leaves,
sweetgum and sycamore, and their rate of decomposition in different flow rates of water. Our
goal was to see if the speed of the stream or the type of the leaf had a greater impact on overall
leaf decomposition. The speeds we are testing are a faster riffle section and a slower pool. Riffles
are the most common type of rough fast water in low gradient channels, and are shallower than
rapids (Hauer, 2017). Pools are areas of a stream where the water tends to be deeper and/or
wider, and the movement is very slow. By having leaf packs in these two different areas, we
measured how the movement of water affects the decomposition of each leaf species over time.
Past studies have shown that different leaf types decompose at different rates, but that almost all
showed significant decomposition in the riffle section of streams, even noting that significant
amounts of mass were lost in only 48 hours (Maloney 1992).

Based on this study and others, as well as knowledge of the river continuum concept, we
hypothesized that the leaves in the riffle regions will show more decomposition than those in the
pool regions due to the fact that faster moving water would break down the leaves faster, and
also more macroinvertebrates will have access to the leaves in these sections. Additionally, we
hypothesized that the sycamore leaf packs will lose more mass than the sweetgum leaf packs due
to the higher surface area of the sycamore leaves and the higher nutrient content.

Methods:

Leaf Collection & Site Location:

In order to conduct our study, we created artificial leaf packs, in two different parts of a
stream over the course of 8 weeks. We collected two different types of leaves, sycamore and
sweetgum. Sycamore leaves were identified by the fact they have 3 points and are fairly large.
The sweetgum leaves on the other hand were identified because they were smaller and had 5
points. These leaves were collected from the ground on the St. Mary’s College of Maryland
campus, specifically at the St. Johns Creek Upstream site, located at 38°11’09.9” N 76°25’37.7”
W.

Leaf pack assembly:

After collecting enough leaves to fill 8 total plastic bags, 4 for each leaf type, we brought them
into the laboratory and separated them by type into metal trays. These trays were then placed into
Model 40GCE-LT laboratory drying ovens for 24 hours, at 70° Celsius. Drying the leaves
ensured a more accurate initial mass measurement. After the 24 hours were complete, we
returned to the lab to create 60 leaf packs, 30 for each leaf type. Each leaf type and flow rate had
three replicates. We weighed out approximately 5 grams of leaves for every pack, recording the
exact weights on an Excel sheet. After weighing, we re-wetted the leaves gently, to allow us to
put them into the mesh bags with as little leaf breakage as possible. Each pack received two
labels, both detailing the treatments, week, and group name. One label was placed inside each
pack, and the other was tied around the top of the pack. Each received two labels instead of one,
to increase the likelihood that each pack could be properly identified.

Placing leaf packs:

Once the leaf packs were completed and labeled, all 48 of them were brought to the St.
John’s Creek Upstream Site, where they will be placed. A FLO-MATE™ model 200 portable
flow meter was then used to determine the two best pool and riffle sites in a section of the
stream. Based on this, we were able to settle on an area with fast flow, and an area with slow
flow. The location of the fast riffle treatment had an average flow of 0.735 m/sec, and the slow
pool had an average of 0.14 m/sec. The packs were then separated by week and flow type and
metal stakes were used to secure the packs into the stream. At the same time as the placement,
our week 0 data was collected. For this, 12 of the leaf packs were simply placed into their
respective positions and then immediately removed, as a control group for our data. They were
still dipped into the water to ensure that the control leaf packs underwent the same environmental
and experimental conditions as the rest of the leaf packs.
Collecting leaf packs:
Every two weeks for 8 weeks, twelve leaf packs, three from each treatment, were
removed from the stream. Before the leaf packs were collected, we measured and recorded the
dissolved oxygen in mg/L and percent, the salinity, and the temperature in Celsius were recorded
using a Pro 2030 Professional Series YSI. Additionally, the flow speed of each area of the stream
was collected using the same model flow meter as before. During each collection date, the packs
were removed from the stream and placed into ziplock bags, and then taken back to the
laboratory.

After collection:
Following leaf pack collection, the packs were immediately brought to the laboratory.
Here, we dumped the leaves and any other contents from the mesh bags into their own tub of
water. We rinsed mud off the leaf particles in these tubs, as well as sorted out any
macroinvertebrates, and removed any sticks or other unwanted items from the bags. The
macroinvertebrates were placed in labeled jars, and the other items were put aside. We then made
a tin foil “boat” and placed the wet leaves inside it. This process was repeated for every leaf pack
removed. Each tin foil boat was then labeled with the leaf type, treatment, week, and our group
name, and then they were placed into the drying ovens once again for 24 hours at 70° Celsius.

Crucibles:
After 24 hours, the boats were removed from the ovens and weighed on an analytical
balance. The weight of the dried leaves was recorded. We then took the dried leaves and placed
them into labeled ziploc bags, and crushed them until the pieces were small and mostly even in
size. An empty crucible was weighed on the Mettler AT261 DeltaRange precision scale, and the
weight was recorded. We then filled the crucible about two-thirds of the way full with the
crushed leaves, and recorded the weight of the full crucible. This was repeated for every leaf
pack, and then we had the TA fire them. Once the crucibles were fired, we reweighed them and
found the ash weight by subtracting the weight of the empty crucible from the weight of the
crucible containing the leaves.
Calculating the loss of organic matter:
To determine the difference in the loss of organic matter over time for the two different
types of leaves and flow speeds, we calculated the percent organic matter of each leaf pack using
this equation:
𝐷𝑊𝑆𝑢𝑏𝑠𝑎𝑚𝑝𝑙𝑒 − 𝐴𝑊𝑆𝑢𝑏𝑠𝑎𝑚𝑝𝑙𝑒
%𝑂𝑀 = 𝐷𝑊𝑆𝑢𝑏𝑠𝑎𝑚𝑝𝑙𝑒
* 100

Then, using the percent organic matter of each leaf pack, we calculated the Ash-Free Dry Weight
of each leaf pack using this equation:
𝐴𝐹𝐷𝑊 = 𝐷𝑊𝑙𝑒𝑎𝑓𝑝𝑎𝑐𝑘 * %𝑂𝑀

Finally, we calculated the percent of Ash-Free Dry Weight remaining in each of the leaf packs,
using the final and initial AFDW. To do this, we used the Ash-Free Dry Weight calculated for the
Week Zero leaf packs as the initial AFDW. This allowed us to determine the amounts of organic
matter lost in each of the leaf packs. The equation used was:

%𝐴𝐹𝐷𝑊 𝑅𝑒𝑚𝑎𝑖𝑛𝑖𝑛𝑔 = ( 𝐹𝑖𝑛𝑎𝑙 𝐴𝐹𝐷𝑊

𝐼𝑛𝑖𝑡𝑖𝑎𝑙 𝐴𝐹𝐷𝑊 ) * 100

Statistical Analysis:
In order to determine the relationship between time passed and amount of organic matter
in the leaf packs, we performed a linear regression on all four of the treatments. These
regressions resulted in R-squared values showing how strong the relationship between time and
amount of organic matter was for each leaf type and speed. We then completed four different
t-tests. We specifically chose t-tests in order to compare only the specific values that we wanted.
The four t-tests performed were: a t-test to compare the decomposition in fast v slow flow speed
for sycamore, a second one comparing fast v slow flow speed for sweetgum, and then a third for
leaf type in slow flow, and finally a fourth for leaf type in fast flow.

Results:

Table 1. Data collected from the fast riffle site in St. Johns Creek, measured at time of leaf
pack collection. Data labeled N/A was unable to be collected due to equipment malfunctions.

Week 0 Week 1 Week 2 Week 3 Week 4

 Dissolved N/A 9.72 6.49 9.68 9.23
oxygen (mg/l)

Dissolved N/A 90.1 63.0 90.8 88.2

oxygen (%)

Salinity (ppt) N/A 0.1 0.1 0.0 0.0

Temperature N/A 12.7 14.4 12.5 13.9

(°C)

Flow Rate N/A N/A N/A 0.53 0.94

(m/sec)

Table 2. Data collected from the slow pool site in the St. Johns Creek, measured at time of
leaf pack collection. Data labeled N/A was unable to be collected due to equipment
malfunctions.

Week 0 Week 1 Week 2 Week 3 Week 4

Dissolved N/A 9.13 6.68 9.53 9.43

oxygen (mg/l)

Dissolved N/A 84.85 63.6 89.2 88.9

oxygen (%)

Salinity (ppt) N/A 0.0 0.1 0.0 0.1

Temperature N/A 12.6 13.4 12.5 13.3

(°C)

Flow Rate N/A N/A N/A 0.14 0.14

(m/sec)

During each day of leaf pack collection, the dissolved oxygen (mg/L), the percent
dissolved oxygen, the salinity, the temperature, and the flow rate of a fast riffle area and a slow
pool area of St. John’s Creek was recorded using a YSI and flow meter. (Table 1, Table 2).
Figure 1. The linear regression of the ln%AFDW remaining over time in weeks of sweetgum
leaves in fast flow, sweetgum leaves in slow flow, sycamore leaves in fast flow, and sycamore
leaves in slow flow calculated across the 4 leaf pack collection weeks. (SG. Fast: P < 0.01; R2 =
0.8223) (SG. Slow: P < 0.01; R2 = 0.407) (Syc. Fast: P < 0.01; R2 = 0.7938) (Syc. Slow: P <
0.01; R2 = 0.2966).

Based on the linear regression, the null hypothesis stating that there is no relationship
between the percent ash-free dry weight remaining and time was rejected. This is due to the fact
that every treatment type shows an inverse linear relationship between the percent ash-free dry
weight and time (Figure 1). The slope of the regression line for sweetgum in the fast flow areas
showed the largest decrease in organic matter over time, at 78%. The second largest decrease
was the sycamore leaves in the fast flow, at 60%. Both of the leaf types in the slow flow areas
showed a lower decrease in organic matter over time, with the sweetgum in the slow areas
decreasing by 20%, and the sycamore in the slow areas only showed a 4% decrease (Figure 1). In
general, the linear regression showed that the leaves in the fast areas decreased much more than
in the slow areas, regardless of leaf type. Additionally, it showed that the sweetgum leaves
decomposed more than the sycamore leaves in their respective treatments.
Figure 2. The mean %AFDW ( ± SE) remaining for four different leaf pack treatments (n = 3):
sweetgum leaves in fast flow, sweetgum leaves in slow flow, sycamore leaves in fast flow, and
sycamore leaves in slow flow.

The sycamore leaves in the fast flow conditions displayed a loss of organic matter that
was 86.38% greater than the loss for the sycamore leaves in the slow conditions (Figure 2; Fast:
mean %AFDW = 11.77; Slow: mean %AFDW = 86.45). This difference in loss of organic matter
between the flow speeds was statistically significant (T-test: p= 0.0149). The sweetgum leaves
showed a similar trend. The loss of organic matter for the sweetgum leaves in the fast areas was
90.82% greater than those in the slow areas (Figure 2; Fast: mean %AFDW = 5.41; Slow: mean
%AFDW = 58.88). This difference was also statistically significant (T-test: p=0.0193).
In the area of slow flow, sweetgum leaves lost 31.89% more organic matter compared to
the sycamore leaves (Figure 2; SG: mean %AFDW = 58.88; Syc: mean %AFDW = 86.45). This
difference between the leaves was statistically significant (T-test: p= 0.03524). In the fast flow
area, there was a difference of 54.07% between the two leaf types (Figure 2; SG: mean %AFDW
= 5.41; Syc: mean %AFDW = 11.77). However, this difference was not statistically significant
(T-test: p= 0.175)
Discussion:
The purpose of this project was to study the decomposition rates of sycamore and
sweetgum leaves in fast and slow sections of the St. John’s stream over the course of 8 weeks.
The project looked at how decomposition rates varied between the leaf types as well as flow
types. The primary focus was on the amount of leaf mass lost, however, macroinvertebrates were
also collected and noted for each leaf type. This research has both small and large-scale impacts,
with both rates of decomposition and macroinvertebrate rates giving insight into the stream’s
health, nutrient levels, and characteristics. We hypothesized that the mass of the leaves in the
riffle regions will be less than those in the pool regions because faster moving water would break
down the leaves faster, and more organisms can reach the leaves in the faster regions.
Additionally, we hypothesized that the sycamore leaf packs will lose more mass than the
sweetgum leaf packs due to the higher surface area of sycamore leaves.
Our first hypothesis is supported by the data collected, and we reject the null hypothesis
which states that there is no relationship between the percent of ash-free dry weight remaining
and time. The leaves in the fast riffle regions lost more organic matter than those in the slow
regions, with sycamore in the fast regions losing over 86% more than in the slow, and with
sweetgum in the fast areas losing over 90% more than those in the slow (Figure 2). The
differences in organic matter lost were statistically significantly different between the fast and
slow treatments for both leaf types (T-test p<0.05). The results of the linear regression also
support this, with the two fast treatments both displaying a larger slope than the two slow
treatments (Figure 1). These results are consistent with the idea of faster moving water leading to
an increase in decomposition. However, our hypothesis that sycamore leaves will lose more mass
than sweetgum was not supported. When collecting the macroinvertebrates we noted that there
were more in the sycamore leaf packs, which aligns with our hypothesis, however, sycamore
leaves did not show a greater loss of mass than sweetgum. In fact, it was the complete opposite.
In the slow flow regions, the sweetgum leaves lost over 31% more organic matter than the
sycamore leaves (Figure 2), additionally, this difference was statistically significant (T-test: p=
0.03524). There was also a difference of over 54% between the leaves in the fast regions (Figure
2), however, this difference was not statistically significant (T-test: p= 0.175).
 Based on this data, we can conclude that the flow rate of water has more of an impact on
the decomposition of leaves in the fast regions, however in the slow regions, where there is less
decomposition due to the moving water, the leaf type has more of an impact.
Our findings of the fast flow being more impactful than the leaf type on the amount of
decomposition are supported by previous research. In one study, the researchers noted that after
only 48 hours all the leaf types had lost substantial mass, except for hemlock leaves which only
lost small amounts. That being said, the study also does note that the leaf type does have an
impact, as does ours. After the first 48 hours, the hemlock lost 7%, but the sugar maple lost 40%
(Maloney 1992). Our data is supported by this study, but one way we could further our study
taking inspiration from this one would be to conduct the same experiment again, but this time
with more types of leaves. Focusing on only sycamore and sweetgum allowed us to compare
those two types found on campus, but expanding to include more leaf types would allow us to
expand the research implications.
One limitation of our results is that we were unable to collect stream data such as
dissolved oxygen and flow rate for either week 0 treatment (Table 1, Table 2). Additionally, for
collection weeks 1 and 2, a flow rate was unable to be collected due to equipment malfunctions
(Table 1, Table 2). This would affect our average flow rate for the two sites. Additionally, the
weather may have drastically changed properties of the stream including but not limited to
increased flow rates after heavy rain. This may be reflected in the experiment or could have also
been lost due to a flow meter malfunction. Having a longer timeframe for the experiment would
help ensure that more weather types were included in the study. Additionally, since we collected
the leaves from the ground and based on our own knowledge, there was no way to 100%
guarantee that every single leaf was of the proper type. Although we were as accurate as
possible, this could affect leaf decomposition levels as well as macroinvertebrate feeding levels.
This study has broader implications in the realm of stream health and nutrients. Amounts
of different decaying leaves and plants can contribute varying nutrient levels to soil and streams
(Bastias et al., 2017), and both nutrient and macroinvertebrate levels are indicators of stream
health, among others. By studying how the trees around the streams affect the health of and who
lives in the streams, we can learn more about the health of streams and of the environment in
general.
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