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Analysis of the Presence of Macroinvertebrates on White Oak (Quercus sp.) Leaves in

Cedar Run Bog in Comparison to Cedar Run Outflow
Sophia Barnes, Cailean Connolly, Theodore Cooke, Joshua Goldman, Gianna Hoheb, Andrey
Khariv, Maximilian Midili
Abstract
Macroinvertebrates, being unable to travel large distances, are used as bioindicators in freshwater
ecosystems. They play major roles as both prey and decomposers, keeping aquatic food chains
intact. Analyzing macroinvertebrates can give clues to the dynamics and structure of a habitat.
White oak (Quercus sp.) leaf packs were placed in two distinct environments to determine
differences in the diversity of macroinvertebrates and how it would affect the contrasting
arrangements of these environments. Two leaf packs were placed in Cedar Run Bog, and the
other two were placed in the Cedar Run Outflow on May 5, 2022. The leaf packs were left for
approximately three weeks until collection on May 26, 2022. It was hypothesized that more
macroinvertebrates would be present in the packs in the outflow in comparison to the bog.
Comparing Simpson’s Dominance and Simpson’s Diversity values, it was determined that the
outflow had a higher diversity of species. The outflow had a diversity value of 0.80 in
comparison to the bog’s value of 0.71. A two-tailed T-test was run and a p-value of 0.315059
was calculated, classifying the results as insignificant.

Introduction
Leaf litter in water bodies are utilized by a variety of organisms as a form of shelter and
food (Konig, Hepp U., Santos, 2014). Different types of leaves, however, play a role in the
variety of macroinvertebrates that will inhabit them. The type of water flow, either lentic or lotic,
also determines the quantity and species of insects that will be present in leaf litter. The presence
of macroinvertebrates can be used as bioindicators for an ecosystem. It is possible to observe the
presence of eutrophication, nutrient flow, decomposition, and types of sediment through the
analysis of macroinvertebrates (National Park Service, n.d.).
Macroinvertebrate species prefer certain types of leaves. The leaf packs in this
experiment are meant to mimic leaf packs found in nature, generally those found near riffles in
lotic streams. The palatability of a leaf, or essentially how “good” it tastes to an organism, is a
factor determining which species prefer to consume it (Stubbs, Blundell, 2013). The leaves of
white oak species generally produce less tannins than that of black or red oak species. Since
tannins produce a bitter sensation to macroinvertebrates, white oak leaves are better in providing
palatability to potential organisms that eat them. The leaves of oak trees in general are also
difficult to break down as a result of their waxy surface and high density. Other trees like maple
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are known to decompose more rapidly since they are easier to tear. Shredder macroinvertebrates,
for example, might prefer white oak leaves due to their palatability and toughness.
The movement of the water body is a huge factor in this experiment, as the mobility of
most macroinvertebrates are dependent on water flow (Leaf Pack Network, n.d.). In order to drift
into a leaf pack, insects require some sort of water movement at the minimum; the motility of an
aquatic insect is limited by its small size. In stagnant water, it would be more likely to find a
smaller number of macroinvertebrates due to a lack of flow limiting them. In flowing water, it
would be expected to find a larger abundance and variety of macroinvertebrates including the
larvae of damselflies, stoneflies, dragonflies, caddisflies, and mayflies.
It is hypothesized that there will be a larger abundance of macroinvertebrates on leaf
packs in flowing water (outflow). There could also be a higher concentration of shredder
macroinvertebrates such as caddisflies and mayflies due to the structure of the white oak leaf.
Since small aquatic insects and larval species are not able to travel large distances, it would be
difficult for them to navigate to a leaf pack on their own (EPA, n.d.). Instead, flowing water
would allow them to be swept downstream into leaf packs. The leaf packs, which consist of
white oak leaves (Quercus sp.), will provide shelter from predators and provide a food source for
the macroinvertebrates. Therefore, it is hypothesized that more macroinvertebrates will be
present in the packs in the outflow versus the bog.

Methodology
Study Site
The tests performed for this study were located in an aquatic habitat containing a bog
running into an outflow. Cedar Run Bog used to be a cranberry bog, owned by Nathaniel Holmes
Bishop. In 1880, Cranberry harvesting became an industry in this area because of experiments
showing that these bogs flooded in order to keep the vines from freezing (Stafford History, n.d.).
The Cedar Run Bog and Outflow are located in the Pine Barrens of Ocean County, NJ (Figure
1). The location of Cedar Run makes it part of the Barnegat Bay Watershed, which comprises
several sub-watersheds and encapsulates all of Ocean County (Ocean County Soil Conservation
District, 2022).
Leaf Sampling
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White oak leaves were chosen because white oaks are commonly found in the area and it
was important to make the leaf packs relate to the environment. White oak leaves (Quercus sp.)
were collected from the Oak-Pine forest near Ridge Road, Stafford, NJ (Figure 2), by sorting
and collecting 120g of leaves into a reusable plastic bag.
Stream Characterization
Cedar Run is a first-order freshwater stream that flows ~1.5 miles through West Creek,
NJ. The stream flows through pine barrens and out into the Barnegat Bay. When the leaf packs
were placed into the stream on May 5, 2022, temperature, conductivity, and DO levels were
tested using a YSI (Figure 6). The bog had a temperature of 14.1 degrees Celsius, a conductivity
rating of 72.9 SPC, and a Dissolved Oxygen level of 9.64 mg/L. The outflow of the stream had a
temperature of 14.3 degrees celsius, a conductivity rating of 73.2 SPC, and a Dissolved Oxygen
level of 9.5 mg/L.
Experimental Design
The collected leaves were equally distributed from the collection bag into four mesh bags
(Figure 4), each receiving 30 grams of leaves per bag (Figure 3) on May 3, 2022. The mesh
bags were joined to a concrete brick through tying twine the bricks and the bags. This was done
so that the bags were weighed down when placed into the lotic water system, and in order to
prevent the bags from being washed downstream. The leaf packs were transported to the site
location and placed into flowing water near the old cranberry bog located along Cedar Run in
West Creek, NJ (Figure 1) on May 5, 2022. Each leaf pack was left untouched for three weeks.
After the three week period, the quantity of macroinvertebrates in each leaf pack was recorded.
Collection and Identification
On May 26, 2022, the leaf packs were collected from the Bog and Outflow (Figure 1),
and taken back to the lab to be analyzed. Each leaf pack was placed in a bucket and opened. The
leaves were then scraped individually to remove any macroinvertebrates within them. The
macroinvertebrates were separated into an ice cube tray and then analyzed and identified under a
microscope using A Guide to Common Freshwater Invertebrates of North America.

Results
There was a larger abundance and diversity of species in the outflow. The predominant
species captured was the flathead mayfly (Heptageniidae sp.), with 57 organisms found in the
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outflow packs (Table 1). 30.8% of the organisms recorded were flathead mayflies, followed by
28.6% isopods, 8.6% net spinning caddisflies, 7.0% midges, and so on (Figure 9). More
shredder macroinvertebrates were found in these packs, including mayflies, isopods, caddisflies,
and midges. The outflow had a Simpson’s Dominance value of 0.20 and a Simpson’s Diversity
value of 0.80 (Table 2). The outflow and bog had an n-value of 22, representing the 22 unique
species recorded between both sites.
The leaf packs in the bog were dominated by isopods with 41 organisms found (Table 2).
Amphipods (Gammarus sp.) followed with 30 organisms (Table 2). 43.2% of the organisms
recorded were isopods and 31.6% were amphipods, totalling almost three-fourths of the species
collected (Figure 10). Other species including damselflies and dragonflies were found in these
packs in much smaller numbers. Similar to the outflow, species with shredder feeding behaviors
dominated the bog leaf packs. The bog had a Simpson’s Dominance value of 0.29 and a
Simpson’s Diversity value of 0.71 (Table 2).
In the outflow packs, there were 18 distinct species consisting of 185 total organisms
(Table 1). The Simpson’s Dominance value of 0.20 (Table 3) represents a well-spread number of
organisms among these species and shows the diversity of this data. In the bog packs, there were
10 distinct species, with 95 total organisms (Table 2). There was slightly more dominance in this
group, with a Simpson’s Dominance value of 0.29 (Table 3). However, this value is still
particularly low, meaning that numbers were still fairly spread out in each species. The T-test
showed that the results are insignificant, with a p-value of 0.315059. This could partly be the
result of the sampling size being too small or a possible unknown factor affecting the data.

Discussion
Combining all of the data, the two main factors affecting the diversity of species in the
two sites were flow rate and pollution. Due to the larger quantity of organisms found in the
outflow compared to the bog, it can be said that the flow of the outflow provided the necessary
transportation for macroinvertebrates considering that they cannot move large distances (Voshell,
J. R., 2003). Amounts of pollution in both sites were determined by the larger amount of
pollution-sensitive species in the outflow, where there were more mayflies, stoneflies, and
net-spinning caddisflies (Table 1). On the contrary, the bog had 0 mayfly and stonefly species.
Thus, it can be concluded that pollution did play a role in the types of species recorded and their
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distribution. The constant in this experiment was the palatability of the white oak (Quercus sp.)
leaf, which made up all four leaf packs in both sites .
The higher flow rate of the outflow provides the environment and the necessary motility
that macroinvertebrates require (Leaf Pack Network, 2020). Since these organisms are mostly
unable to move large distances, flow is a key factor in transporting macroinvertebrates (EPA,
2021). The flow from the outflow was thought to have provided more transportation for these
species, thus providing support as to why there might have been more organisms in the outflow
leaf packs. This is contrary to the bog, where the primary environment was more lentic. As a
result, less organisms would be able to be transported into the leaf packs. This is supported by
the spread out distribution of species in the outflow, whereas the bog had slightly more
dominance with almost half of the organisms being isopods (Figure 10). The outflow could have
also been a more desirable environment for organisms like mayflies and stoneflies since they
have a preference for clinging on to leaves in streams with higher flow rates (Stumpf, S.,
Valentine-Darby, P., & Gwilliam, E., 2015). This also coincides with the fact that the outflow had
more riparian area, like the bank of the stream. This type of riparian zone is necessary for
macroinvertebrate reproduction cycles, especially since adult macroinvertebrate species like
insects inhabit wooded areas near water (König, R., Hepp, L. U., & Santos, S., 2014).
Additionally, the 57 flathead mayflies (Heptageniidae sp.) found in the outflow can be
used to compare the levels of pollution in the outflow versus the bog (Table 1). Contrary to this
result, the bog hosted 0 flathead mayflies (Table 1). According to the Pollution Tolerance Index
(PTI), mayflies are ranked under Class 1 in terms of sensitivity to pollution. Their high
sensitivity to pollution can indicate that the bog potentially consisted of larger quantities of
pollution than the outflow (EPA, 2021). 1 perlodid stonefly was also found in the outflow versus
0 found in the bog, further representing this idea since stoneflies are also in Class 1 (Table 1).
There were 24 more scuds (Gammarus sp.) in the bog, however, which are ranked under Class 2
of the PTI (Table 1). This ranking indicates a higher tolerance to pollution than Class 1
organisms, portraying the differing pollution levels in the Cedar Run Bog and Cedar Run
Outflow. In general, the outflow was home to more sensitive macroinvertebrate species, hinting
at the pollution levels in both ecosystems.
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Figure 1: map of testing site: Figure 2: Students collect

Blue pinpoint - site 1 (bog) white oak leaves from an
Yellow pinpoint - site 2 (outflow) oak-pine forest near Ridge
Road, Stafford, NJ.

Figure 3: 30 grams of Figure 4: Each mesh bag packed and

leaves were measured and shaped similarly to each other.
put into four mesh bags.

Figure 5: YSI testing of Figure 6: 2 leaf packs placed at Figure 7: 2 leaf packs placed
the outflow. site 1 (bog). at site 2 (outflow).
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Figure 8: Macroinvertebrates
from each pack were identified
in the lab

Figure 9: Pie chart of the collected species of the outflow

Figure 10: Pie chart of the collected species of the bog

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Table 1: Total species list from both leaf packs

Table 2: Simpson’s Dominance and Simpson’s Diversity values for the bog and outflow

Conclusion
The hypothesis was supported, as there was a larger quantity and diversity of species
found in the outflow. This was shown by comparing Simpson's Dominance and Simpson’s
Diversity values between both the Cedar Run Outflow and Cedar Run Bog. Visually juxtaposing
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the tables and pie charts, it is evident that there were more species and a larger abundance of
organisms in the outflow leaf packs. This could be the result of the outflow’s faster flow rate,
providing transportation and easier access to the leaf packs. The bog, however, did host many
organisms, but less of them were insect larvae. This represents the motility of macroinvertebrates
and their subsequent use as bioindicators. The results also showed that pollution might have
played a role in the types of species found since there were more pollution-sensitive species.
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