Sediment Toxic Effects
on Aquatic Micro Life
By Patrick Ebbert, Central Catholic, PJAS 2008
Introduction



Algal food production could be inhibited by a
variety of pollutants in the marine ecosystem.
Some of this pollution can even be found in the
runoff of our own parking lots into streams and
rivers.
Algae populate the oceans as well as freshwater
systems abundantly and provide a basis for the
entire world’s food web due to their
photosynthetic abilities.
Two different species of algae (Chlamydomonas
and Euglena) were tested with two different
parking lot sediment samples to examine the
effects.
Sediment Effects on Marine Life

1.
2.
There are two properties of sediment that
could be potentially dangerous to aquatic
life.
The runoff into water ecosystems can cloud the
water, making it harder for light to pass through
and therefore harder for autotrophs to provide
themselves with sufficient energy via
photosynthesis.
The contents of the sediment, possibly abundant
with chemicals leaked from cars or other
industrial locations, could have some adverse
affect on the organisms because of its foreign
molecular structure.
Runoff Properties


Runoff from parking lot sediment can contain
many chemicals harmful to life.
May contain herbicides, pesticides,
polychlorinated biphenyls (PCBs), polycyclic
aromatic hydrocarbons (PAHs) and more.
Location of Sediment and Possible Impacts



Sediment A was taken
from a school parking lot.
Chemicals from fuel or
rubber particles from tires
could have possibly leaked
into the ground.
Sediment B was taken
from the driveway of a
public park, containing
similar particles to
Sediment A.
Both of these zones were
located near woodland
streams containing aquatic
life.
Characteristics of Algae
Euglena





Have one large flagellum
Typically 35-55um long
Over 100 species
Survive best in rich organic
waste
Contain many chloroplasts
Chlamydomonas





Have two flagella
Typically 10-30um wide
Eyespot helps them
gravitate toward light
Found in calm waters and
damp soil
Contain only one
chloroplast
Purpose

The purpose of this experiment was to see
how various concentrations of parking lot
sediment affect the growth of euglena and
chlamydomonas.
Hypotheses

Null: The growth rates of the algae
exposed to parking lot sediments will not
vary significantly from the controls.

Alternative: The growth rates of the algae
will vary due to different concentrations of
parking lot sediment.
Methods and Materials
Basic Experimentation


Four sets of each euglena
and chlamydomonas were
put into test tubes with
various concentrations of
rock sediment and kept
under favorable lighting
conditions
Spectrophotometer set at
blue-green wavelength
(430nm) was used to
measure absorbency of
algae every 3 days for 2
weeks.
Materials
 Test tube rack (1)
 Test tubes (56)
 200µl micro-pipette (1)
 10ml macro-pipettes (3)
 5ml macro-pipettes (3)
 Spectrophotometer (1)
 Permanent marker (1)
 48ml of Euglena
 48ml of Chlamydomonas
 160ml of spring water
 Daphnia magna (At least
64)
 Mini pipette (1)
 Sediment from 4 different
parking lot locations
Procedure
1.
2.
3.
Arranged 48 test tubes in test tube rack; pipetted 2ml of
Euglena into 24, and 2ml of Chlamydomonas into 24.
Pipetted sediment and spring water into each tube as
displayed in chart on following slide.
Absorbance readings at 430nm were recorded on days 1,
4, 7, 10, and 13.
Procedure (cont.)
Sed. A:
0%
Sed. A:
10%
Sed. A:
40%
Sed. B:
0%
Sed. B:
10%
Sed. B:
40%
Spring
Water
3ml
2.5ml
1ml
3ml
2.5ml
1ml
Algae
2ml
2ml
2ml
2ml
2ml
2ml
Sediment
0ml
0.5ml
2ml
0ml
0.5ml
2ml
Total
5ml
5ml
5ml
5ml
5ml
5ml
*Repeat this table 4 times for Euglena and 4 times for
Chlamydomonas
Algal Growth Curve
0.9
Algal Growth Curve
0.8
Average Absorbance at 430nm
0.7
Euglena A-0%
Euglena A-10%
0.6
Euglena A-40%
Euglena B-0%
0.5
Euglena B-10%
Euglena B-40%
0.4
Chlamy A-0%
Chalmy A-10%
0.3
Chlamy A-40%
Chlamy B-0%
0.2
Chlamy B-10%
Chlamy B-40%
0.1
0
Day 1
Day 4
Day 7
Time
Day 10
Day 13
More Graphical Comparisons
Sediment Effects on Chlamydomonas Populations
Sediment Effects on Chlamydomonas Populations
0
0%
10%
0
40%
0%
10%
40%
-20
Sediment B
Sediment A
-60
Sediment B
-80
Perecent Change in Absorbency at 430 nm
Sediment A
-40
-100
-150
-100
-200
-120
Concentration of Sediment
-250
Concentration of Sediment
Sediment Effects on Euglena Populations
Sediment Concentration Effects on Euglena Populations
250
150
200
Sediment B
Sediment A
Sediment B
50
0
0%
10%
-50
40%
Sediment A
Percwent Change of Absorbency at 430nm
100
Percent Change of Absorbency at 430nm
Percent Change in Absorbency at 430nm
-50
150
100
50
0
0%
10%
-50
-100
-150
Concentration of Sediment
-100
Concentration of Sediment
40%
ANOVA Statistical Analyses
2-Factor Comparison of Sediment on Chlamydomonas
ANOVA
Source of Variation
SS
df
MS
F
P-value
F crit
Sample
136.3308333
2
68.16542
1.344136
0.285716084
3.55455715
Columns
30.60041667
1
30.60042
0.603401
0.447371702
4.4138734
Interaction
28.01083333
2
14.00542
0.276169
0.761841588
3.55455715
912.8375
18
50.71319
1107.779583
23
Within
Total
ANOVA Statistical Analyses (cont.)
2-Factor
2-FactorComparison
ComparisonofofSediment
Sedimenton
onEuglena
Euglena
ANOVA
Source of Variation
SS
df
MS
F
P-value
F crit
Sample
1172326.603
2
586163.3
2.773092
0.089159558
3.55455715
Columns
110215.7067
1
110215.7
0.521422
0.479512608
4.4138734
Interaction
264199.7433
2
132099.9
0.624954
0.546505665
3.55455715
Within
3804757.025
18
211375.4
Total
5351499.078
23
Results and Conclusions



The null hypothesis that the euglena would not
vary due to sediment concentrations was
accepted (P-value >.05).
Also, the null hypothesis that the chlamydomonas
would not vary due to sediment concentrations
was accepted (P-value >.05).
In conclusion, the alternative hypothesis was
rejected because there was no significant
difference between either of the sediments when
compared to their effects on both euglena and
chlamydomonas.
Limitations and Extensions




The sediment could have slightly clouded the test
tube solution, giving false readings of
absorbency.
The algae also may have been an unhealthy
sample (as indicated by the decline in population
density)
More trials could have been used, or maybe
trying the readings on more than one
spectrophotometer.
Other algal samples, including natural aquatic
samples could be tested.
Further Research: Daphnia Magna


For an external
experiment, the
water-flea Daphnia
Magna was tested
with the rock
sediment in only one
trial.
Daphnia were put in
tubes of water and
sediment and
compared by survival
percentage
Daphnia 1st-Trial Results & Conclusions




Daphnia were compared in eight
total tubes, four with Sediment A,
four with Sediment B.
The concentrations of sediment 120%
used were 0%, 10%, 20%, and
40%.
100%
To the right is a graph showing
the survival percentage of each
tube of daphnia over a period of 80%
one week.
The Daphnia exposed to no
sediment appeared to have a
60%
slower decline rate compared to
the ones with 10-30%
concentration.
40%
This experiment could be
expanded further to test more
effects of sediment on aquatic
20%
animal groups rather than plant
groups such as algae.
Sediment Effects on Daphnia Survivorship
Perecntage of Daphnia Alive

A-0
A-10
A-20
A-30
B-0
B-10
B-20
B-30
0%
Day 1
Day 2
Day 3
Day 4
Time
Day 5
Day 6
Day 7
References








“Chlamydomonas reinhardtii”. Wikipedia.org.
<http://en.wikipedia.org/wiki/Chlamydomonas_reinhardtii>. Dec. 16, 2007.
“About Chlamydomonas”. Chlamy.org. <http://www.chlamy.org/info.html>. Jan. 7,
2008.
“Euglena”. Wikipedia.org. <http://en.wikipedia.org/wiki/Euglena>. Jan. 21, 2008.
“Structure of a Euglena”. Infovisual.info.
<http://www.infovisual.info/02/001_en.html>. Jan. 21, 2008
Newcombe, C.P., Macdonald, D.D. “Effects of Suspended Sediments on Aquatic
Ecosystems.” North American Journal of Fisheries Management.
<http://afs.allenpress.com/perlserv/?request=getabstract&doi=10.1577%2F1
5488675(1991)011%3C0072%3AEOSSOA%3E2.3.CO%3B2&ct=1>. Dec. 4,
2007.
Dr. John Wilson, Biostatistician, University of Pittsburgh.
Clare, John, PhD. “Daphnia”. Caudata.org. <http://www.caudata.org/daphnia/>.
Jan. 23, 2008.
“Contaminated Sediment in Water”. EPA.gov.
<http://www.epa.gov/waterscience/cs/>. Dec. 4, 2007.