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A Rural Environment Compared to an Urban Environment:

Effect of Human Environment Type on the Benthic Macroinvertebrate Diversity of Rivers
By Chloe Leonard

Purpose
Benthic macroinvertebrates are an essential part of an aquatic ecosystem, but many of these invertebrate
communities can easily be stressed by a change in their natural environment. What with the widespread increase of
urban and agricultural activity, aquatic ecosystems may be increasingly pressured by these human influences.
I have always been interested in wildlife and the environment, and the biology and ecological role of invertebrates
have especially intrigued me. Therefore, I aimed to investigate and compare the water quality of two different river
systems – a rural water system and an urban water system – by examining the biodiversity of benthic
macroinvertebrate communities in each. This was done by analyzing three variables: the richness, or total number of
different taxa in a collected organism sample, the abundance of these organisms, and the different tolerance levels of
these taxa.

Background
The physical and chemical state of a river is impacted by its drainage area, which includes all land that collects and
drains water to the river and its smaller branching streams (United States Geological Survey 2018). Also drained is the
collection of nutrients, pollutants, and sediments from the surrounding area, which in varying amounts have direct or
indirect negative impacts on the aquatic organisms that are limited to their habitat. This especially includes benthic
macroinvertebrate communities, since certain types of these organisms are more susceptible to change than others
(Zimmerman 1993).
A susceptibility to change refers to the organisms’ tolerance levels, which estimates how capable they are of surviving
in an environment with conditions (i.e. dissolved oxygen levels) less than optimal to their survival preference
(Regional Aquatics Monitoring Program 2008). By this idea, organisms that are less tolerant to unfavorable conditions
will be less in numbers, if at all present. A decrease in the number of taxa, or organism types, in an ecosystem results
in less species richness. Oppositely, organisms that are tolerant of these conditions will persist and even thrive in an
open ecological habitat. They can spread to available spaces not inhabited by the otherwise intolerant species, and in
result there is a lack of evenness in the different populations and there is clearly a dominant set of certain organisms
(Covich, Palmer, Crowl 1999).
The tolerance levels of benthic macroinvertebrates vary depending on the animal’s taxonomic group. A biotic index
shows the tolerance levels of certain groups by numerical values and uses them to calculate an approximate value for
biodiversity (Zimmerman 1993); however, these values differ per different type of biotic index since many index
values apply to only specific conditions and regions of the world. For this study, the Stream Quality Monitoring (SQM)
method developed by the Ohio Department of Natural Resources was used for identifying certain taxa and for
applying tolerance values to them. The SQM method divides a total of 20 different taxa into three groups of tolerance
levels: Group 1, the intolerant organisms, including all mayfly larvae (Ephemeroptera), caddisfly larvae (Tricoptera),
stonefly larvae (Plecoptera), water penny larvae (Psephenidae), dobsonfly larvae (Megaloptera), riffle beetles
(Elmidae), and gilled snails (Pleuroceridae); Group 2, containing somewhat tolerant organisms, including all other
beetle larvae (Coleoptera), damselfly larvae (Zygoptera), dragonfly larvae (Anisoptera), cranefly larvae (Tipulidae),
crayfish (Cambarus), scuds (Amphipoda), sowbugs (Isopoda), and clams (Unionoida); and Group 3, containing tolerant
organisms including blackfly larvae (Simuliidae), midge larvae (Chironomidae), leeches (Hirudinea), pouch snails
(Physidae), and aquatic worms (Annelida, Platyhelminthes, Nematomorpha) (Zimmerman 1993). Any organism in a
sample that did not fit into these taxonomic groups were omitted from being counted into the data.
Additionally, the number of organisms collected per taxa was considered: organism types that were found in a sample
but in numbers less than 10 were considered “rare”, while organisms ranging between 10 and 99 were “common”,
and organisms that were found in numbers of at least a hundred were “dominant” (Environmental Protection Agency
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Office of Water 1997). For the further and better organization of data, the SQM method was adapted to assign certain
values to the organism numbers that were identified per taxa. Number values were used by the EPA Volunteer
Stream Monitoring: A Methods Manual, and they assign certain values to the rarity of taxa by their tolerance
groupings (see Table 3.1). This specifically allowed for a better quantitative comparative analysis between the
different values taken from the two different rivers.
Riffle microhabitats provide ideal conditions for many benthic macroinvertebrate species, since oxygen is easily mixed
below the surface of water that is broken by submerged or exposed rocks (RAMP 2008). These locations therefore
provide an excellent source for collecting a variety of sensitive organisms in need of higher oxygen levels, since they
would unlikely be found in microhabitats with less dissolved oxygen. Furthermore, riffle areas provide easier and
more consistent data collection than other microhabitats, since they provide better control of certain variables such
as depth and water flow that affect the types of organism communities that live there (Zimmerman 1993).
All organisms were collected from riffle areas by benthic stream monitoring techniques. For each trial, a sample was
taken from three different riffle sites of an area approximately 1 m², and no site was used more than once. Samples
were collected from both rivers within a 200 m area lengthwise, to limit collection within a certain area of both rivers
and to not extend to a possibly different set of physical or biological characteristics. For each riffle site, the
temperature, depth, and velocity of the riffle were measured so that values could be compared; total averages
amounted to very similar values for the two river locations. Both rivers were visited a total of five times each, totaling
to fifteen different riffles sampled per river.

Research Question
How does a rural environment versus an urban environment affect the biodiversity ratings of benthic
macroinvertebrate communities from riffle microhabitats in an aquatic river system?

Hypothesis
If a river within an urban environment is compared with its biodiversity to a river within a rural environment, then the
biodiversity ratings for benthic macroinvertebrate communities in an urban river will be less diverse, and therefore
less healthy, than the values collected from a rural river. This is because an urban environment potentially holds a
greater variety and amount of pollution sources than a rural environment, what with its overall greater density of
human activity.

Variables
– Independent Variable – n/a
The main cause of change in biodiversity for a river is, in this case, human activity. This includes the introduction of
pollutants such as pesticides or heavy metals, sediments, or nutrients such as phosphorus or nitrates that in excess
amounts may cause eutrophication, or oxygen-loss in an aquatic system (EPA Office of Water 1997). In this
investigation, the presence or abundance of these substances were not measured, but rather the effects they could
give on a system’s biological diversity were.
– Dependent Variable – biodiversity rating of benthic macroinvertebrates from riffle habitats
– Constants –

 Rivers are within the same watershed (Upper Scioto region) – Since the watershed includes all land that
drains water into a central river system, rivers of the same watershed will more likely share similar
characteristics, such as temperature, precipitation, and pH, all of which that can affect biotic factors.
 Both rivers were visited on the same day – Weather conditions such as precipitation will temporarily affect
the physical characteristics of a river, and consequentially the ability to collect data (EPA Office of Water
1997). Visiting the collection sites on the same day prevents any irregularities between the data collection of
both rivers.
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 Data was collected within the same season (mid-summer) – Different types of benthic macroinvertebrates
respond to the seasons in different ways, having greater or less appearance depending on the time of year
(EPA Office of Water 1997). Collecting all data within the same season keeps a consistency in the variety of
available organisms.
 Same type of microhabitat collected from – Since all collection was restricted to riffle areas, only organisms
expected to be found in riffles were part of the data. This allows easier organization of data, since only one
population type per river was considered.
 Same area used for data collection for both rivers – Riffles were accessed within a 200-meter area
lengthwise along both rivers from a point selected on a map; it is possible that locations further upstream or
downstream of these areas could contain different physical or biological characteristics that could skew that
river’s data.
 Same number of riffles (3) collected from per river and per collection day – This keeps a constant on the
cumulative measurement of collected organisms, since a relationship is expected to appear between the two
different river types.
 Same amount of time used for collecting organisms per riffle site – Substrate stirring was timed to 80
seconds at each site of collection. This allowed a constant in the possible number of burrowing organisms
collected.

Safety
Supplies: Sunscreen, bug spray, closed-toed water shoes, bandages (in case of injury), drinking water for hydration
The areas along both rivers that were visited are part of public property within parks and required no prior permission
to be accessed from any land-owner. Furthermore, both areas were suggested by employees of the Ohio Department
of Natural Resources for being ideal locations of data collection from riffles (Szymanski, Doherty 2018).
An assistant was used whose entire purpose was to hold the seine net during the collection of organisms. Data was
collected from riffles by the comfort of the collector. Any area in a river that was considered too deep, rapid, or
otherwise too harsh by the collector was avoided. Likewise, if weather conditions were considered too harsh, data
was not collected that day.
Data was collected by proper benthic monitoring procedure as suggested by experienced individuals and by
documentation from the Ohio Department of Natural Resources (ODNR 2010) and the US Environmental Protection
Agency (EPA 1997). All collected organisms were returned to the stream after identification.

Materials
For collecting organisms: Seine net (1x1 m, 1/16 inch mesh), 2 medium-sized buckets (with handles for easier
carrying), measuring cup or container (for scooping/ pouring water and easy carrying)

For measuring riffle characteristic data: water-proofed thermometer (± 1C), plastic ruler (cm), water-resistant string
(measured 1 meter in length), fishing bob, timer/ stop watch
For identifying organisms/ recording: 2 white plates (shallow, slightly bowl-shaped for holding organisms in water;
white for easy identification), small plastic shovel/ spoon, magnifying glass (for easier identification of organisms),
waterproof notebook, pen, clipboard and paper (for recording data), organism ID key (if needed), calculator

Procedure
1. Three riffle locations of different types (differing depths or velocities) were planned out to collect data from.
2. With the organism-collection and measuring materials, the furthest riffle downstream was approached; this
was so the riffle habitat and organisms were not disturbed in order to prevent the possibility of data being
influenced.
3. Benthic macroinvertebrates were collected using stream monitoring techniques.
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a. A 1x1 meter riffle area was chosen, and an assistant positioned the seine net just downstream from
it.  The seine net was angled so that its front remained in contact with the bottom substrate while its
back remained well above the water.
b. Within the 1x1 meter riffle area, rocks were removed with any clinging invertebrates brushed off into
the net. Some rocks were placed on the bottom of the net so that the net remained in full contact to
the substrate, which prevented benthic macroinvertebrates from flowing under the net. The
remaining rocks were placed outside the sampling riffle (they were returned after data collection was
finished).
c. Within the 1x1 meter riffle area, the bottom substrate was stirred by shuffling the feet; this allowed
burrowing invertebrates within several centimeters of the substrate to be dislodged and carried into
the net by the flow of the stream. This was done by starting from the far end of the riffle, upstream,
working toward the net so that the benthic organisms were less likely to flow outside the net. The
substrate was stirred for 80 seconds, until the area was thoroughly mixed.
d. While the net held the collected organisms, the rocks were quickly removed from the bottom lip of
the net, and the net was carefully lifted from the stream. With the help of the assistant, the net was
carried to the bucket horizontally and folded to prevent organisms from falling out.
e. All, if any, non-invertebrates collected (i.e. fish) were returned to the stream.
f. The net was positioned vertically into the bucket so that the side with the collected organisms was
folded inward. New debris that clearly had no organisms on them were picked off by hand. The
measuring cup was used to wash as much of the contents of the net as possible into the bucket with
water from the stream.
g. After washing was finished, the assistant held the net up horizontally, so that the side holding any
remaining clinging organisms or debris was directed upward. With the waterproof notebook and
pen, all remaining organisms and taxa on the net were made note of and tallied. This was done for
the efficiency of time, since some organisms, e.g. caddisfly larvae, were capable of strongly clinging
to surfaces and were difficult to wash off with the water. Furthermore, with less water accumulated
in the bucket, there was generally less time required for later organism identification.
h. After all organisms on the net were recorded, the net was returned to the stream and carefully and
thoroughly washed of all remaining organisms back into the water.
i. Physical characteristics of the riffle were measured. This raw data was later used for comparing the
average characteristics between each stream.
i. The thermometer was completely submerged for 2 minutes. The temperature was recorded
in the waterproof notebook.
ii. The depth (cm) of the riffle from the deepest part was measured with the ruler.
iii. The velocity of the riffle was measured. This was done by first measuring out a meter
distance with the string between two physical points. The fishing bob was dropped and
collected after it traveled the designated distance while the stopwatch measured the time.
This was repeated for a total of 3 trials.
j. Steps a. through i. were repeated for the next two riffles, while moving upstream. All three samples
were composited into the same bucket.
4. The collected macroinvertebrates were identified.
a. The filled sampling bucket and the other empty bucket were set next to each other.
b. With the measuring cup, water was scooped from the sampling bucket onto the white plate. The
organisms and taxa were identified and counted, and the numbers were tallied on paper. The plastic
shovel/ spoon was used to scoop out certain organisms onto the second white plate if needed.
c. After as many organisms were identified on the plate as possible, the contents of the plate were
poured into the empty bucket so that the counted organisms were organized as such and were kept
in water.
d. These steps were repeated until all the contents of the sampling bucket were completely gone
through.
5. After as many organisms were identified as possible, they were returned to the stream.
6. This data collection process was repeated for the other remaining river.
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Results
Table 1.1: Total Organisms Collected Per Tolerance Level

Rural River Urban River

Somewhat
Somewhat
Day TOTA Day Intolerant Tolerant Tolerant TOTAL
Intolerant Tolerant Tolerant
L
1 299 17 49 365 1 213 11 47 271
2 378 31 83 492 2 216 9 77 302
3 678 19 86 783 3 131 12 113 256
4 677 31 52 760 4 331 11 136 478
5 717 44 28 789 5 212 21 122 355
TOTAL 2749 142 298 3189 TOTA 1103 64 495 1662
L

Table 1.2: Number of Different Taxa Collected

Day Rural River Urban River

1 14 13
2 13 7
3 13 8
4 12 8
5 13 10
Average: 13 9.2
SD: 0.71 2.39

Table 1.3: Number of Taxa according to Rarity (number of organisms collected)

Rural River Urban River

Rare Common Dominant Rare Common Dominant
Day (1-9 org.) (10-99 org.) (100+) Day (1-9) (10-99) (100+)
1 8 5 1 1 9 3 1
2 6 6 1 2 3 3 1
3 5 6 2 3 4 3 1
4 4 6 2 4 3 4 1
5 5 6 2 5 5 4 1
Average: 5.6 5.8 1.6 Average: 4.8 3.4 1
SD: 1.52 0.45 0.55 SD: 2.49 0.55 0

Immediate Observations:
As shown in Table 1.1, there was an overall greater number of organisms collected from the rural river. This is
noteworthy since the same collecting procedures were used for both environments. It can be assumed by this fact
alone that there is some difference in the physical or chemical conditions of the two rivers — the conditions in the
rural river are able to support more organisms than the conditions in the urban river. Furthermore, the urban river
had notably more tolerant-class organisms than the rural river. This is significant since the urban river had nearly half
the total collected organisms of the rural river. The urban population sample had a significantly greater portion of
tolerant organisms when compared to the quantity of intolerant organisms, which suggests a more prominent
dominance of tolerant organisms in that environment.
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According to Table 1.2, there is a notable difference in the number of different taxa collected. The rural river had an
average of 13 different taxa collected with little variability between each day, with a standard deviation value less
than 1. The urban river had an average of 9 different collected taxa with greater variability between each day, with a
standard deviation of approximately 2. This shows that, while the urban environment can hold nearly as many
different organism types as the rural river, it is possible that the population of benthic macroinvertebrates in the
urban river has more abundance of organisms in certain types of taxa rather than having more of an even spread.

Table 1.3 shows the number of taxa that were in rare, common, or dominant occurrence. As stated by the EPA, “It is
never ideal for any given type of organism to dominate a sample, and… it is best to have a wide variety of organisms
including a few pollution-tolerant individuals” (EPA Office of Water 1997). According to this information, an aquatic
system should ideally consist of organisms that are “common”, since it would suggest a more equal spread of these
organism types rather than having a taxon that is overly abundant or too few. All collections from both rivers
contained a dominant taxon, mayflies, and the rural river also had dominant caddisflies. Both rivers had somewhat
inconsistent “rare” taxa, as shown by the standard deviation values which are both greater than a value of 1. In
contrast, both rivers had consistent “common” taxa, with standard deviation values less than 1. This suggests a
prominence of these organisms within both rivers, but not to an overwhelming extent in the overall benthic
macroinvertebrate populations. It must be noted, however, that the rural river has a greater average of “common”
taxa than the urban river.

Data Processing and Analysis

 Percentage:
Value ∈sample
%= ∙100
Total
Percentage of total intolerant organisms collected out of the total rural river sample:

717
%= ∙ 100 = 86.20%
3189

Urban
Rural River:
River: Percentage
Percentage of
of Organism
Organism Types
Types  Average:
9%
4%
30% Intolerant
Intolerant
Somewhat
Somewhat
Tolerant
Tolerant
Tolerant
Tolerant
A=
∑ of numbers
Number of terms
4%
66%

86%

Average for the number of taxa collected for the rural river:

14 +13+13+12+13
A= = 13
5
 Standard Deviation:
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Equation for standard deviation for a sample, where x is the value of the data set, μ is the mean of the
data set, and n is the number of data points in the sample.

∑∣ x−μ∣ ²
SDsample =
√ n−1
Standard deviation of different taxa from the rural river:
2 2 2 2 2
SDsample = ∣ 14−13∣ +∣13−13∣ +∣13−13∣ +∣12−13 ∣ + ∣13−13 ∣ = 0.71
√ 5−1

 Chi Square Test for Homogeneity:

While there is clearly a difference in the number of organisms collected in the two rivers, it is difficult to
determine whether there is a significant difference between the spread of numbers of the two river’s
tolerance groups. This can be determined by a chi square test for homogeneity.

In the test for homogeneity, two variables were tested: the quantitative values of the populations from
the rural river and the urban river. Two hypotheses were tested, as shown:

Null hypothesis: both rivers have an even distribution of organism types

Alternative hypothesis: there is a significant difference in the distribution of organism types between the two rivers.

DF = degrees of freedom
Degrees of Freedom: DF =(r – 1)∙( c – 1) r = total rows
c = total columns
Er,c = expected frequency counts
nr = total number of observations
Expected Frequency Counts: (nr ∙ nc) from population r
Er,c = nc = total number of observations at
n
treatment c
n = total sample size
Χ² = chi squared
Test Statistic: ( ¿ , c−Er , c )2 Or,c  = observed frequency value
Χ² = Σ[ ] Er,c = expected frequency value
Er , c

Table 2.1: Total Observed Organisms

Somewhat
Intolerant Tolerant Tolerant TOTAL
Rural River 2749 142 298 3189
Urban River 1103 64 495 1662
TOTAL 3852 206 793 4851

Table 2.2: Total Expected Organisms

Somewhat
Intolerant Tolerant Tolerant
Rural River 2532 135 521
Urban River 1319 70 271
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Degrees of Freedom: Expected Frequency Counts for Rural Intolerant:

(3189 ∙3852)
DF = (2 – 1) ∙ (3 – 1) = 2 Er,c ¿ = 2532
4851

After the expected values were calculated, they were plugged into the equation for the test statistic:

(2749−2532) ² (142−135) ² (298−521) ² (1103−1319)² (64−70)² ( 495−271)²

Χ² ¿ + + + + +
2532 135 2532 1319 70 271
Χ² = 18.6 + 0.36 + … + 185.15
Χ² = 335.66
The p-value used was 0.05 (5%). According to the critical values chart for chi square, the value of use is 5.991. The
value calculated from the collected data was 335.66. Therefore, the statement can be made:
335.66 > 5.991
Since the calculated value is greater than the critical value, the null hypothesis was rejected. Instead, the alternative
hypothesis was accepted: there is a significant difference in the distribution of organism types between the two
rivers.
While this does not determine the biodiversity ratings, it proves that the rural river has a different spread of
organisms than the urban river, and vice versa. This shows that, because the two rivers have different drainage areas
and environments, they subsequently contain different aquatic environments that can support their own range of
benthic macroinvertebrates.

 Biodiversity Ratings:
The biodiversity or stream quality rating is calculated from certain weighting variables, depending on
locational region. These were the weighting variables used for this investigation:

Table 3.1: Weighting Variables

Somewhat
Organism Count: Intolerant Tolerant Tolerant
Rare (1-9) 3.0 2.2 1.2
Common (10-99) 3.6 2.4 1.1
Dominant (100+) 3.3 2.0 1.0
(EPA Office of Water 1997), (ODNR 2010)

The formula to calculate biodiversity rating for the method used is:

BR = ∑ [(value for taxa tolerance per rarity) x (weighting variable)]

For example, if 4 different intolerant taxa were present but each counted to values less than 10, they are considered
“rare” with a rating factor of 3.0. The number of taxa, 4, is multiplied by that weighting factor. The biodiversity rating
for the rural river on Day 1 was calculated as such:

( 4 ∙3 )+ (2 ∙ 3.6 ) + ( 1∙ 3.3 ) + ( 2 ∙2.2 ) + ( 1 ∙2.4 ) + ( 2 ∙1.2 ) + ( 2 ∙1.1 )=33.9

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Table 3.2: Biodiversity Ratings

Day Rural River Urban River
1 33.9 29.5
2 34.3 16.5
3 34.6 17.7
4 31.4 18.7
5 33.6 23.1
Average: 33.56 21.2
SD: 1.27 5.32

According to the calculated values in Table 3.2, the rural river had all biodiversity scores greater than 30, suggesting
very good water quality, while all values in the urban river had values greater than 15 but less than 30, suggesting
good or fair water quality. The rural river had an average score of 33.56 with a standard deviation of 1.27, while the
urban river had a lesser average score of 21.2 with a greater standard deviation of 5.32. This larger range of ratings is
due to the different numbers of taxa that were collected per day.

A greater variance in the calculated values for the urban river indicates that, while a certain taxon was counted one
day, it was not seen often or again on other days. This discrepancy in taxa found may suggest an unequal distribution
of benthic macroinvertebrate types, since if there were a prevalence of certain organisms in the river, they would
likely be found in multiple riffle areas.

Biodiversity Rating Per Day

40
35
Biodiversity Rating

30
25
20
15
10
5
0
0.5 1 1.5 2 2.5 3 3.5 4 4.5 5 5.5
Day

Rural River Urban River

Linear (Urban River)

Conclusion
The purpose of this investigation was to determine if there was a difference between the benthic macroinvertebrate
communities of a rural river environment versus an urban river environment; and, if there was a difference, if the
macroinvertebrate diversity ratings were significantly stronger in either population sample. Furthermore, it was
predicted that the rural river environment contained a greater biodiversity rating than the urban river environment,
and therefore contained more ideal water quality. According to the data, this hypothesis was supported.

The value that was calculated for chi square was significantly greater than the critical value for 5%, which rejected the
null hypothesis. This adds proof that, even though either river had a different total value of collected organisms, the
two rivers had a different spread in the types of organisms that were collected. As seen from the raw data, the urban
river provided a significantly greater quantity of tolerant organisms when compared to the rural river. According to
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the concept of the biotic index, if there is a greater number of organisms that are capable of surviving in poorer water
conditions, this suggests that the water quality of the stream is less than ideal (Zimmerman 1993). While there were
less tolerant organisms collected than intolerant organisms in the urban river, there was overall a greater percentage
of tolerant organisms in the urban river.

The percentages calculated for the total organisms in the three tolerance groupings show that the benthic
macroinvertebrate samples collected from the rural environment are dominated mainly by intolerant invertebrates.
While the urban river likewise had a significant number of intolerant invertebrates, there was also a greater
proportion of tolerant invertebrates to the total. This fact alone would propose a more even spread of different
organism types in the urban river, except the number of different taxa collected suggest otherwise.

The average values of the number of taxa collected per river show that the rural river consistently provides a higher
quantity of taxa than the urban river. The higher standard deviation value for the urban river demonstrates the
unequal spread of collected taxa per day, which suggests that an inconsistency in the type of organisms collected
relate to the lack of occurrence of these certain taxa. This may add to the idea that resilient organism types are more
prevalent because there is less prevalence of more susceptible organisms that would otherwise fill a space in the
ecological web.

Both rivers show a similar trend in taxon rarity; the rarity values show that there is at least one “dominant” taxon, a
mostly consistent number of “common” taxa, and an overall greater number of “rare” taxa. It must be noted,
however, that benthic macroinvertebrate populations may change depending on the season, and that certain seasons
may yield an abundance of certain kinds of benthic macroinvertebrates (EPA Office of Water 1997). This can be an
explanation to the overall strong appearance of a taxon. The overall larger number of taxa in the “rare” category
shows that there were many taxa that made only an occasional appearance within the composited samples taken; the
organisms of these taxa might be outcompeted by others of the same benthic macroinvertebrate population, are
restricted in numbers by environmental factors, biologically require less numbers to sustain a population of taxon, or
that less were collected due to the error of the collectors – for example, it is possible that some taxa may prefer
certain areas of a riffle where the collectors did not go to. The first two possibilities refer to some limiting factor on
the organism’s prosperity, which can be caused by poor water quality or competition from more successful
organisms. A consistency in “common” taxa suggests that there is a balance in the numbers of these organisms; there
is no establishment of dominance for the organism type in the microhabitat, and an even spread of different
organisms contribute to the invertebrate population’s evenness and overall biodiversity. The rural river, however,
clearly shows a greater number of “common” taxa, which suggests that, not only does the stream sustain a richness in
the number of taxa present, but there is evenness in the distribution of these organisms.

The calculated biodiversity ratings tentatively reflect the water quality of the stream. As shown by the values for each
day, there are consistently better ratings for the rural river when compared to the urban river. The average of the
ratings shows that there is about a numerical difference of 12.46 between the ratings of the two rivers, which is about
a third of the average rating for the rural river, or 33.56%. This is a significant difference between the two rivers. This
fact, as well as the other supporting information from the tables, graphs, and calculations, give confirmation that the
river in the rural environment has a greater biodiversity in its benthic macroinvertebrate communities than the river
in the urban environment, since the urban environment has many sources within the water basin that can drain
harmful pollutants; the rural environment, therefore, seems to deliver less pollution, and that would overall
contribute to a better stream quality.
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Evaluation and Improvement

The drainage area of each river is of different size. Depending on what is within the drainage area will likely influence
river quality, which may affect biodiversity ratings. According to the United States Geological Survey, the point
investigated on the rural river has a water basin of 695.2 km² while the point investigated on the urban river has a
water basin of 300.9 km². A water basin that is larger can include more area that water and nutrients can accumulate
in a central channel, possibly providing a more suitable environment for a greater array of organisms which may be
partially reflected on the greater number of organisms collected from the rural river. While the urban river basin
contains half the land of the rural river basin, 35% of this land, or 106.2 km², is used by urban activity, while the rural
river basin is 7.7% urban activity, or 53.4 km². This urban activity is significantly more dispersed, however, since most
of the land in the rural river basin is used for agricultural activity, which is likewise a general source of pollution. It is
difficult to say, therefore, whether the overall size of the two river basins have a significant impact on the collected
data and results.

For further consideration is the SQM method that was used for identifying certain types of taxa. This method, as
stated by members of the Ohio Department of Natural Resources, is not as accurate as other biotic indexes. The SQM
method was chosen since it allowed for rapid identification of broader taxa, since the invertebrates were to be
identified in the field and returned to the stream as soon as possible; this method does not require the use of a
microscope, which is a necessary component when identifying taxa such as Ephemeroptera to the family or genus
level. Furthermore, by the investigation’s requirements, visiting both rivers on the same day required raw data to be
collected as efficiently as possible. For more accurate and trustworthy results for the calculated biodiversity ratings,
the Hilsenhoff Family Biotic Index can be used, as it provides tolerance levels for many taxa in the family level of
organization (Zimmerman 1993); this method however would require organisms to be preserved and taken to the lab
for identification, which would not accept the standards for the IB Policy on Animal Experimentation.

Because invertebrates were identified in the field as efficiently as possible, there is the likelihood that there is some
error in the counting or identifying of organisms; either the same organisms were counted more than once or missed,
or certain organisms were mistaken to be of a different taxon (beetle larvae, for example, looking very similar to
certain caddisfly larvae). This would result in specific organism counts to be slightly off than the reality.

According to the EPA, a river will have a change in flow rate and depth within several days after rain, since benthic
macroinvertebrates can wash downstream. Collection Days 3 and 5 took place within this time span of rain, and both
rivers were observed to have a change in flow rate and depth, more so in the urban river. Consequentially, there were
the least amount of collected organisms on Day 3 for the urban river. If data were collected outside of at least a 72-
hour timespan of rain for each collection day, there would be no chance of outliers from this variable.

Benthic macroinvertebrates, like other organisms, require nitrogen and phosphorous for vital living processes. Having
more or less quantities of these nutrients will affect the quality of living for aquatic organisms; a lack of nutrients will
inhibit life functions, while excess amounts can result in eutrophication and limit the quantities of dissolved oxygen,
which is likewise vital for the functioning of aquatic organism (Khan Academy 2018). Nitrogen and phosphorous levels
were not measured in this investigation. Other substances that may affect the quality of water (e.g. pesticides, heavy
metals, sediments) were also unmeasured. Data collected for the presence of these variables would enhance the
quantitative comparison between the two rivers, since it would compare the direct chemical makeup of the different
aquatic habitats.
 12

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