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BENTHIC MACROINVERTEBRATE

SAMPLING AS INDICATORS FOR


WATER QUALITY

Grant Hackworth
STOCKTON UNIVERSITY

Benthic Macroinvertebrate sampling as indicators for water quality

Contents
Abstract ......................................................................................................................................................... 1
Introduction .................................................................................................................................................. 2
Materials ....................................................................................................................................................... 3
Methods ........................................................................................................................................................ 4
Experiment Preparation ............................................................................................................................ 4
Leaf Pack Placement ................................................................................................................................. 4
Leaf Pack Removal and Analysis ............................................................................................................... 4
Results ........................................................................................................................................................... 5
Discussion...................................................................................................................................................... 6
Appendix I: Maps .......................................................................................................................................... 7
Historical Map ........................................................................................................................................... 7
ArcGIS Map ............................................................................................................................................... 8
Appendix II: Tables and graphs ..................................................................................................................... 9
Appendix III: Photographs........................................................................................................................... 19
References .................................................................................................................................................. 28

Abstract
This report was very valuable in the observation to the quality for a freshwater ecosystem
within close-proximity to an urbanized area, a zoo, and a city park. The aims were to study the
pollution sensitive macroinvertebrates, total biological richness, biodiversity, and the abiotic
water quality. Initial observations that the raceway, river, and lake would be in fair or poor health
seemed to hold true throughout the experiment. The lack of species richness and biodiversity, as
well as low EPT indices did not favor good or excellent habitat conditions. Water quality was
classified as good in regards to nitrogen, pH, and dissolved oxygen levels, but had very high
phosphate levels. Due to the number of other students involved there could have been error in
sampling or the samples did not pick up the pollution that was causing the lack of biodiversity
among the macroinvertebrate community. Ultimately the experiment established a starting point
for more research of similar nature to be carried out and will now have a base of indices and
standards to be compared to.

Benthic Macroinvertebrate sampling as indicators for water quality

Introduction
The purpose of this experiment is to determine the health of the waterbodies using
macroinvertebrate counts within the benthic layer of the waterbody by method of leaf pack
sampling. Benthic macroinvertebrates are bottom dwelling organisms such as aquatic insects,
insect larvae, crayfish, clams, snails, aquatic worms, and diatoms They are categorized by the
four ecological roles they play in a freshwater system; shredders, filter-collectors, grazers, and
predators. These benthic macroinvertebrate species are an integral part of the freshwater
ecosystem by eating algae and other nutrients and decomposing leaf litter and other materials
while also serving as a food source to birds, fish, reptiles, and amphibians in the ecosystem. The
benthic community also helps transport energy downstream in a watershed (Vannote et al., 1980)).
See Figure 1
Benthic macroinvertebrates serve as indicators for water quality because they are always
living within the water system and allow researchers to study the long-term health of the stream.
There have been many studies conducted using these organisms over the last few decades which
has allowed for researchers to thoroughly document the different pollution tolerances of the
many different species. Each species reacts to the biotic and abiotic factors in the system with
different sensitivity. The EPT index is based on three pollution intolerant species;
Ephemeroptera, Plecoptera, and Trichoptera. The index is used as ecological indicator when
determining the impacts to freshwater sources like streams and lakes (Rios, 2006). There are
many types of EPT indices for evaluating water quality and for this report three were employed
on the collected data. EPT/C index is a ratio of the amount of Ephemeroptera, Plecoptera, and
Trichoptera individuals found versus the amount of Chironomidae (Mandaville, 2002).
Chironomidae are also known as midges and have high pollution tolerance so finding high
amounts of this insect is a sign ecosystem disturbance. The ratio is a quantitative example for
how many pollution intolerant flies there are compared to the amount of high pollution tolerant
midges. The EPT percentage index is a measurement of the total amount of EPT species found
versus the total number of macroinvertebrates found in the sample. The last EPT index is
calculated simply by adding the amount of families found out of each species and the total
number of different families corresponds to a numerical rating. All three of these tests illustrate
the species richness in these very sensitive insects (Mandaville, 2002).

Benthic Macroinvertebrate sampling as indicators for water quality


There is a total of five indices that make up one of the metrics for the DEPs use of
benthic macroinvertebrates when examining water quality. The EPT index involving the amount
of families counted and the EPT percentage make up two of the criteria. The other indices
include percent dominance, taxa richness, and the biotic index. Percent dominance is the ratio of
the dominant species found in the sample compared to the total number of individuals found in
the sample. Taxa richness is simply the number of taxonomic families found in the sample and
correlates to the overall biodiversity of the stream or lake system. The biotic index is an index
where each species or family is given a score associated with certain chemical and physical
requirements in the water. The number of individuals in each species is multiplied by its
respective pollution tolerance value and the sum of each calculation is taken. The index score is
then derived by dividing the sum of pollution tolerance values and dividing them by the total
numbers of individuals in the sample (Tzilkowski, 2008). A well-balanced biological community
is a good indicator of a healthy stream and can give a better understanding of the long-term
health for the system. However, the overall biological health of a waterbody is best studied
through implementation of the above indices in conjunction with tests for pH, nitrogen,
phosphorus, and dissolved oxygen levels. Therefore, understanding the ratios of pollution
tolerant/sensitive organisms and overall biodiversity can better illustrate the water quality.

Materials

YSI meter

Bucket

15 Mesh Bags

Pair of overall waiter boots

Nylon rope

6 Dissection trays

0.45 kg of leaves from survey area

2 pairs of Tweezers

Sharpee

Magnifying glass

Rebar/Stones

Large fine mesh sifter

Camera

3 Sorting buckets

Zip ties

Macroinvertebrate Identification sheet

6 Test Tubes

Pollution Tolerance Identification sheet

Scissors

Dissecting microscope

Benthic Macroinvertebrate sampling as indicators for water quality

Methods
Experiment Preparation
Collected leaves from the nearby area and set them out to air dry before weighing them.
Weighed 30 grams of leaves for each bag to ensure all bags are the same weight, and placed the
leaves in their respective mesh bag. Tied off each mesh bag full of leaves with a zip tie and
secured a piece of 6-10 inch nylon rope for attachment at the sample location. Attach waterproof
labels/markers and assign each a number to be used for site map identification purposes.

Leaf Pack Placement


Selected locations of the stream where the leaf packs can be weighed down and tied off
with the pack facing upstream, riffle areas of the stream work best. Due to the canals soft sandysilt bottom, rebar was hammered into the bottom and each bag was secured using the attached
rope and zip-ties. The leaf packs placed in the Cohansey River were weigh down by being
attached to a small rock and tied off to nearby trees for easy location and removal. Sunset Lake
leaf packs were note placed by me directly and therefore not included in this section. Each leaf
pack was placed in the stream making sure that the most surface area possible is exposed to the
current and the packs are completely submerged and securely tied down and weighted. Packs
remained in the water for 3 weeks and were checked once halfway through to ensure they were
still secured. At time of placement and at the halfway mark, pH, nitrogen, phosphorus, dissolved
oxygen, and temperature were all sampled for each location. The date, time, and ambient air
temperature, were also documented the day the packs were placed and removed. All locations
were marked with assigned leave pack numbers to the constructed site map of the sampling area.

Leaf Pack Removal and Analysis


Leaf packs were removed three weeks after placement at their respective sites and time
intervals. Water temperature, air temperature, dissolved oxygen content, and pH level were
documented the day of the packs removal. No packs were lost for any reason, however, river
pack 3 was out of the water upon arrival due to significantly reduce water level from low tide
and is a potential source of error. Each pack was removed by first placing a few inches of stream
water in the zip-lock bag and readying it for when the bag was detached. Attached strings or zip
ties were cut and leaf packs were gently lifted to the surface and slid into zip lock bag. Each bag
was then put into the cooler with ice for transport. This ensures all the organisms are kept alive
for easier identification. A bucket worth of stream water was collected to use in the rinsing and
4

Benthic Macroinvertebrate sampling as indicators for water quality


sorting process to not shock the macros. Bring back to lab and document each leaf packs number
and location on data sheets. Sort through each leaf pack separately, removing every
macroinvertebrate you find and keeping tally of how many and what species using the
identification sheets. Created an Excel sheet with all the individuals counted at their respective
sample locations and ran the indices using excel. Used the pollution toleration worksheet to
determine the quantity of each species per tolerance level and determine the quality of water with
the results. Apply results to EPT %, EPT/C indices, biotic index, biodiversity distribution, and
overall macroinvertebrate ratios and water quality ratings.
(Procedures from LPN Resources, 2015)

Results
Cohansey River had a total EPT of 1.2%, the raceway with 0.3%, and Sunset lake with
3%, all well below the unhealthy urban standard (Figure 2). For EPT index involving number of
families counted only one EPT species was found in each site location. The raceway samples
only contained Trichoptera, Sunset Lake contained Ephemeroptera, and the Cohansey River
contained Plecoptera (Figure 2). For the EPT/C ratio, the Cohansey River had the highest with
9.1%, Sunset Lake with 6.7%, and the raceway with just 1.4% (Figure 2). Taxa richness for the
Sunset Lake and the raceway both saw a total of seven different species (Figure 4 & 6) The
Cohnasey River had the most overall species counts with 12 (Figure 5). The overall biodiversity
was weighted towards Chironomidae, Amphipoda, and Oligochaeta with these dominant species
making up 80% of the overall sample population (Figures 7-9). Comparison of EPT species,
other insect species, and non-insect species also show very low numbers of the highly sensitive
EPT species throughout all three waterbodies (Figures 14-16). Pollution tolerance values for the
Cohansey River were between 6.7-7.3 receiving a poor-quality rating at all four sample sites
(Figure 10 & 13) Pollution tolerance values for the raceway were between 6.6-7.1 receiving a
poor-quality rating at all six sample sites (Figure 11 & 13) Pollution tolerance values for Sunset
Lake were between 6.1-6.7 receiving a fair-quality rating at four sample sites and poor-quality at
the fifth site (Figure 12 & 13). When applying these indices to the metric criteria for rating water
quality, the canal scored a 12, the Cohansey River scored a 15, and Sunset Lake scored a 9 all
receiving a moderately impaired classification (Figures 17-19).
5

Benthic Macroinvertebrate sampling as indicators for water quality

Discussion
Analysis of the experiment show that the water quality in the Bridgeton raceway,
Cohansey River, and Sunset Lake are moderately to severely impaired. Similar studies have
shown that healthy forested waterbodies contained close to 50% Ephemeroptera, Plecoptera, and
Trichoptera (EPT) percentage index, whereas unhealthy urban waterbodies were less than 5%.
(Johnson, et al., 2013). The Cohansey River, raceway, and Sunset lake all contained less than 5%
making them all well below the unhealthy urban standard. The other indices involving EPT were
all very low finding only one species of Ephemeroptera, Plecoptera, or Trichoptera in the entirety
of each sample location. These low percentages signify that overwhelming abundance of
Chironomidae populations in comparison to all three EPT species populations. Healthy streams
have even distribution of all four of these species, so when the benthic community is
disproportionately weighted in favor of Chironomidae species it indicates environmental stresses
in the ecosystem (Mandaville, 2002).
There is a clear lack of species richness illustrated by the overall biodiversity observance
in both individual species and EPT, other insect and, non-insect species. Percent dominance
indices agree with this fact as well with the top three dominate species all be highly tolerant
worm and crustacean species. In fact, 80% of all the individuals counted were either midge,
scud, or aquatic worm. The biotic index showed fair water quality with a substantial amount of
organic pollution. The Cohnasey River and raceway scored poor water quality values in all
samples. Sunset Lake scored fair water quality values for the first four sample locations and poor
for the fifth. These values with the lakes average measured nitrogen levels being 3 mg/L and the
average phosphate concentration of 2.0 mg/L there is evidence to conclude the abundance of
biological pollution in the raceway and the river as well. Further supporting this conclusion is
that the dissolved oxygen and pH levels were not indicating poor water quality, therefore there
must be another source causing poor health within the benthic community (Mandaville, 2002).
Many studies of riparian vegetation have shown the lack thereof being the reason for an
influx of nutrients and suspended matter. In fact, riparian vegetation is essential to the health of a
freshwater ecosystem in a few ways. The first role it plays is to act as a buffer for the pond by
absorbing any nutrients or contaminants that would otherwise run directly into the water.
6

Benthic Macroinvertebrate sampling as indicators for water quality


Secondly, they provide shade to help maintain a stable temperature for the aquatic ecosystem.
While the last benefit is providing organic materials for the macroinvertebrates to use in their
energy cycle of a freshwater ecosystem (Rios et al., 2006). Future studies should take samples
upstream from the lake as well as during spring and summer for the best overall understanding of
the benthic community. Potential sources of error include misidentification of species, missed
individuals in sorting process, improper handling of samples from locations to the lab, and river
3 leaf pack being out of water upon discovery for removal.

Appendix I: Maps
Historical Map Historical map of Bridgeton park, raceway, Sunset Lake and Cohansey River.

(Map courtesy of Jim Bergman)

Benthic Macroinvertebrate sampling as indicators for water quality


ArcGIS Map ArcGIS map of macroinvertebrate sample locations at Bridgeton, NJ

(Map made using layers from Bureau of GIS, 2016)

Benthic Macroinvertebrate sampling as indicators for water quality

Appendix II: Tables and graphs

Figure 1

(Vannote et al., 1980)

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 2. Numbers of individuals found at all three locations, with calculated EPT percentages and EPT/C
index

Figure 3. Graphical representation of total biodiversity in species found over all three locations

10

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 4. Individuals found in across all six sample locations in the Bridgeton raceway.

Figure 5. Individuals found in across all four sample locations in the Cohansey River.

Figure 6. Individuals found in across all five sample locations in the Sunset Lake.
11

Benthic Macroinvertebrate sampling as indicators for water quality

Figure 7. Biodiversity representation of Bridgeton Raceway.

12

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 8. Biodiversity representation of the Cohansey River.

Figure 9. Biodiversity representation of Sunset Lake.

13

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 10. Bridgeton raceway pollution tolerance value chart for biotic index
Pollution
Macroinvertbrate

Species

Midge
Scud
Snail
Aquatic Earthworm
Ribbon Worm
Leech
Springtail
Damselfly
Caddisfly
Sowbug
Black Beetle
Stonefly
Crab
Mayfly
Hellagrammite
Dragonfly
TOTAL

Chironomidae
Amphipoda
Gastropoda
Oligochaeta

Canal 1

Canal 2

Canal 3

Canal 4

Canal 5

Canal 6

Tolerance Value Count TTV Count TTV Count TTV Count TTV Count TTV Count TTV

Nemertea

Hirudinea
Collembola

Zygoptera
Trichoptera
Isopoda
Coleoptera
Plecoptera
Decopoda
Ephemeroptera
Megaloptera
Anisoptera

6
6
7
8
8
10
5
7
2.8
8
4.6
1
8
3.6
2
4

20
18
3
27
0
0
0
3
0
0
0
0
0
0
0
0
71

Total Tolerance Value(TTV)/Count = 6.845

120
108
21
216
0
0
0
21
0
0
0
0
0
0
0
0
486

34
62
6
41
0
0
0
0
0
0
0
0
0
0
0
0
143

204
372
42
328
0
0
0
0
0
0
0
0
0
0
0
0
946

6.615

42
34
20
50
0
1
0
5
0
0
0
0
0
0
0
0
152

252
204
140
400
0
10
0
35
0
0
0
0
0
0
0
0
1041

6.849

14
18
6
25
0
4
0
3
0
0
0
0
0
0
0
0
70

84
108
42
200
0
40
0
21
0
0
0
0
0
0
0
0
495

7.071

19
21
7
47
0
1
0
1
2
0
0
0
0
0
0
0
98

114
126
49
376
0
10
0
7
5.6
0
0
0
0
0
0
0
688

7.016

6.656

Figure 11. Cohansey River pollution tolerance value chart for biotic index
Pollution
Macroinvertbrate

Species

Midge
Scud
Snail
Aquatic Earthworm
Ribbon Worm
Leech
Springtail
Damselfly
Caddisfly
Sowbug
Black Beetle
Stonefly
Crab
Mayfly
Hellagrammite
Dragonfly
TOTAL

Chironomidae
Amphipoda
Gastropoda
Oligochaeta
Nemertea

Hirudinea
Collembola

Zygoptera
Trichoptera
Isopoda
Coleoptera
Plecoptera
Decopoda
Ephemeroptera
Megaloptera
Anisoptera

River 1

River 2

River 3

River 4

Tolerance Value Count TTV Count TTV Count TTV Count TTV
6
6
7
8
8
10
5
7
2.8
8
4.6
1
8
3.6
2
4

12
34
12
17
9
7
5
5
0
13
3
3
0
0
0
0
120

Total Tolerance Value(TTV)/Count = 6.823

14

72
204
84
136
72
70
25
35
0
104
13.8
3
0
0
0
0
818.8

9
19
5
25
0
9
0
7
0
1
0
0
0
0
0
0
75
7.333

54
114
35
200
0
90
0
49
0
8
0
0
0
0
0
0
550

2
5
3
2
0
1
0
0
0
0
0
0
0
0
0
0
13
6.846

12
30
21
16
0
10
0
0
0
0
0
0
0
0
0
0
89

10
16
2
5
0
2
0
1
0
0
0
0
6
0
0
0
42
6.786

12
26
6
16
0
0
0
4
0
0
0
0
0
0
0
0
64

60
96
14
40
0
20
0
7
0
0
0
0
48
0
0
0
285

72
156
42
128
0
0
0
28
0
0
0
0
0
0
0
0
426

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 12. Sunset Lake pollution tolerance value chart for biotic index
Pollution
Macroinvertbrate

Species

Midge
Scud
Snail
Aquatic Earthworm
Ribbon Worm
Leech
Springtail
Damselfly
Caddisfly
Sowbug
Black Beetle
Stonefly
Crab
Mayfly
Hellagrammite
Dragonfly
TOTAL

Chironomidae
Amphipoda
Gastropoda
Oligochaeta
Nemertea

Hirudinea
Collembola

Zygoptera
Trichoptera
Isopoda
Coleoptera
Plecoptera
Decopoda
Ephemeroptera
Megaloptera
Anisoptera

Lake 1

Tolerance Value Count TTV


6
6
7
8
8
10
5
7
2.8
8
4.6
1
8
3.6
2
4

17
0
0
0
0
0
0
4
0
0
0
0
0
1
0
0
22

102
0
0
0
0
0
0
28
0
0
0
0
0
3.6
0
0
133.6

Total Tolerance Value(TTV)/Count = 6.073

Lake 2

Lake 3

Lake 4

Count TTV Count TTV Count TTV


10
18
0
0
0
1
0
1
0
0
0
0
0
0
0
0
30
6.167

60
108
0
0
0
10
0
7
0
0
0
0
0
0
0
0
185

10
12
0
0
0
1
0
7
0
0
0
0
0
1
1
0
32
6.144

60
72
0
0
0
10
0
49
0
0
0
0
0
3.6
2
0
197

2
2
0
0
0
0
0
3
0
0
0
0
0
1
0
0
8
6.075

Figure 13. Biotic Index grades for all sampled locations

15

12
12
0
0
0
0
0
21
0
0
0
0
0
3.6
0
0
48.6

Lake 5
Count TTV
6
0
0
0
0
2
0
1
0
0
0
0
0
0
0
1
10
6.7

36
0
0
0
0
20
0
7
0
0
0
0
0
0
0
4
67

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 14. Bridgeton Raceway ratio of EPT species, other insects, and non-insects.

Figure 15. Cohansey River ratio of EPT species, other insects, and non-insects.

16

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 16. Sunset Lake ratio of EPT species, other insects, and non-insects.

Figure 17. DEP metric for water quality using the five indices for all three locations.

Category
Canal
Taxa Richness
EPT Index
% Dominance
EPT %
Biotic Index
Total Score

River
3
0
6
0
3
12

Lake
6
0
6
0
3
15

3
0
3
0
3
9

Figure 18. Water Quality Metric score criteria


Metric
Taxa richness (# of species)
EPT Index (# of Ephemeroptera,
Pleceoptera, and Trichoptera
families)
Percent Dominance (%
contribution of individual in
dominant species to the total # in a
Percent EPT (% of total individuals
from Ephemeroptera, Pleceoptera,
and Trichoptera families)
Biotic Index (Pollution tolerance
value)

17

Metric Score
6
3
>10
10-5

0
4-0

>5

5-3

2-0

< 40

40-60

>60

>35

35-10

< 10

<5

5-7

>7

Benthic Macroinvertebrate sampling as indicators for water quality


Figure 19. Water Quality Metric Classification
Non-Impaired = 24-30
Moderately Impaired = 9-21
Severely Impaired = 0-6

Figure 20. Average nitrogen and phosphorus sampled

(Courtesy of the Sunset Lake water quality group)

18

Benthic Macroinvertebrate sampling as indicators for water quality

Appendix III: Photographs


(all photos taken by Grant Hackworth)

Photo 1

Preparation of leaf packs

Photo 2

Finished leaf packs ready for placement


19

Benthic Macroinvertebrate sampling as indicators for water quality


Photo 3

Canal sampling location 1

Photo 4

Canal sampling location 3


20

Benthic Macroinvertebrate sampling as indicators for water quality


Photo 5

Canal sample location 4

Photo 6

Canal sample location 5


21

Benthic Macroinvertebrate sampling as indicators for water quality


Photo 7

Canal sample location 6

Photo 8

River sampling site 3 at first placement


22

Benthic Macroinvertebrate sampling as indicators for water quality


Photo 9

River sampling site 3 at retreival, low tide is approximately 5 feet lower and the leaf pack is shown here
being out of water, source of error

Photo 10

River sample location 4


23

Benthic Macroinvertebrate sampling as indicators for water quality


Photo 11

In lab leaf pack sorting and identification set-up.

Photo 12

Damselfly (left), Scud (center), Midge (right)

24

Benthic Macroinvertebrate sampling as indicators for water quality

Photo 13

Sorted benthic macroinvertebrates from leaf pack. Scud (Amphipoda) bunched on the left and right,
midge (Chironomidae) bottom and top center and brown casings shown, aquatic earthworms
(Oligochaeta) mid center, and three damselflies (Zygoptera).

Photo 14

Riparian beetle (Coleoptera) found in River 1 leaf pack bag


25

Benthic Macroinvertebrate sampling as indicators for water quality

Photo 15

Ribbon worm (Nemertea) found in River 1 leaf pack bag

Photo 16

Close-up of sowbug (Isopoda) top, damselfly (Zygoptera) in middle, and scud (Amphipoda) on bottom
26

Benthic Macroinvertebrate sampling as indicators for water quality

Photo 17

Two tiny crabs (Decopoda)

27

Benthic Macroinvertebrate sampling as indicators for water quality

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