Download DerenzoFINAL PJAS PRESENTATION

Tap Water Intrusion
Effects on Algal Populations
Anthony DeRenzo
Grade 9
Pittsburgh Central Catholic High School
Surface Runoff
• Is part of the water
cycle and describes
the water that flows
over a land surface.
• It can come from
precipitation and flow
into these two
systems.
Types of Surface Runoff
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Petroleum
Pesticides
Many organic and inorganic chemicals
Silt
• Fertilizers
• Tap water
Tap Water
• Water directly drawn from
a faucet.
• Used in every day
activities such as
plumbing, washing
machines, and
dishwashers.
• Contains a wide variety of
contaminants.
• Goes through a
purification process to set
each contaminant at a
harmless standard.
Common Components in Tap Water
Calcium
9.84 mg/L
Iron
0.0034 mg/L
Potassium
1.36 mg/L
Magnesium
1.72 mg/L
Manganese
0.0002 mg/L
Sodium
8.76 mg/L
Sulfur
4.45 mg/L
Silicon
0.77 mg/L
Strontium
0.56 mg/L
Zinc
.0022 mg/L
Chlorine
0.2 mg/L
Fluoride
0.8 mg/L
Trace Contaminants in Tap Water
cryptosporidium
asbestos
giardia lamblia
barium
legionella
lead
coli forms (including fecal
coli form and E. coli)
mercury
carbofuran
nitrate and nitrite
bromate
atrazine
chlorite
benzene
antimony
many more various organic
contaminants
Eutrophication
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Eutrophication is the process
by which a body of water
becomes enriched in dissolved
nutrients that stimulate the
growth of aquatic plant life.
A eutrophic body of water
promotes the growth of algae
due to increased nutrients
which promotes population
growth.
This reflects the purpose
because it is being tested to
determine if tap water or tap
water and sodium nitrate would
have an effect on the nutrients
in the water. This would then
affect the growth of algae if
there is a negative effect.
Sodium Nitrate
• A colorless, odorless
crystalline compound that
has chemical properties
that resemble potassium
nitrate.
• Formed by a chemical
reaction of sodium
chloride and nitric acid.
• Used in the production of
explosives, fertilizers, and
potassium nitrate. It also
is used to preserve
meats.
Relationship
• People make use of sodium nitrate by growing
plants with fertilizers.
• People make use of tap water through sewage
systems, plumbing, and many other activities.
• A fraction of these two substances undoubtedly
enter natural water sources.
• There are numerous species of macroscopic
and microscopic organisms that could be
affected by runoff that contains either tap water
or tap water + sodium nitrate.
Algae
• All algae lack organic
structures such as roots,
leaves, seeds, and
flowers.
• All algae are
photosynthetic and
usually inhabit damp
places or bodies of water.
• Algae play very important
roles in aquatic ecology.
• There are two major
groups of algae;
prokaryotic and
eukaryotic.
Chlamydomonas
• Single celled green algae
that are common in soil
and freshwater.
• Have two flagella; one
visible that is used for
gliding and swimming, or
metaboly and one that is
not usually not visible.
• Typically can grow in the
light on a medium of
inorganic salts and.
Euglena
• There are many different
shapes of euglena such as
oval, ribbon-shaped, and
cylindrical.
• They also have two flagella;
one that is used to swim and
one that is not visible.
• Have a red pigmented eyespot
that is part of a light receptive
system.
• They reproduce through binary
fission and vary from 20 to 300
nanometers.
Purpose
• Test tap water and a mixture of tap
water + sodium nitrate on two types of
algae: Chlamydomonas reinhardtii and
Euglena sp.
Null and Alternative Hypotheses
• Null- tap water will not significantly affect the population
survivorship of Chlamydomonas reinhardtii or Euglena
sp.
• Null- tap water with sodium nitrate will not significantly
affect the population survivorship of Chlamydomonas
reinhardtii or Euglena sp.
• Alternative hypothesis- the addition of different
concentrations of tap water and tap water with sodium
nitrate will significantly (adversely) affect the algal
models.
Materials
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science kit educator spectrophotometer
orange test tube rack
64 13 X 100 Borosilicate culture tubes
5 ml pipette
100W incandescent light bulb
68 ml of a population of chlamydomonas reinhardtii
68 ml of a population of euglena
16 ml of spring water
8 ml of tap water
1 ml of sodium nitrate
syringe filter
Procedure 1
• Tap water was sterile filtered with a 0.22 micron
syringe filter.
• The following ingredients were added to the
culture tubes to create a total of 5 ml in each
tube: tap water, spring water, sodium nitrate, and
the two types of algae (chart on next page). The
ingredients were added at different
concentrations and were made into 8 separate
groups for each type of algae.
Procedure 2
Algae
2 ml
2 ml
2 ml
2 ml
2 ml
2 ml
2 ml
2 ml
Spring
Water
3 ml
2.5 ml 2 ml
.5 ml
2.95
ml
2.45
ml
1.95
ml
.45 ml
Tap
Water
0 ml
.5 ml
1 ml
2.5
0 ml
.5 ml
1 ml
2.5 ml
Sodium
Nitrate
0 ml
0 ml
0 ml
0 ml
.05 ml
[.1%]
.05 ml
[.1%]
.05 ml
[.1%]
.05 ml
[.1%]
Total
5 ml
5 ml
5 ml
5 ml
5 ml
5 ml
5 ml
5 ml
Procedure 3
• 4 equivalent tubes were created for each
treatment. 64 tubes were created in all.
• The 64 tubes were placed in the orange rack
and set on a table 1.5 meters below the 100W
incandescent light bulb. The light bulb was on
the entire time.
• The tubes were left in the rack for a total of 17
days.
• The absorbance at 430 nm was recorded every
other day. There were 9 recordings total.
Tap Water and Nitrate Effects on
Chlamydomonas
Absorbance at 430 nm
0.700
0.600
0% TW
0.500
10% TW
0.400
20% TW
50% TW
0.300
0% TW+N
0.200
0.100
10% TW+N
20% TW+N
0.000
50% TW+N
0
2
4
6
Time (days)
8
10
% Change in
Absorbance at 430 nm
Tap Water and Nitrate Effects on Chlamydomonas
(% Change from Day 1 to Day 9)
20
15
10
5
0
0% TW 10%
TW
20%
TW
50% 0% TW 10% 20% 50%
TW
+ N TW + N TW + N TW + N
Concentration of Tap Water
Average % Change
The Effects of Tap Water With and Without Sodium
Nitrate on Chlamydomonas
20
15
No Sodium Nitrate
10
Sodium Nitrate
5
0
0% Tap
Water
10% Tap 20% Tap 50% Tap
water
Water
Water
Concentration of Tap Water
ANOVA
Chlamy.
Source of Variation
SS
df
MS
F
P-value
F crit
Sample
428.084
3
142.6947
14.05985
1.7E-05
3.008787
Columns
188.083
1
188.083
18.53201
0.000243
4.259677
10.68706
3
3.562354
0.351002
0.788772
3.008787
Within
243.5782
24
10.14909
Total
870.4322
31
Interaction
Tap Water and Nitrate Effects on Euglena
Absorbance at 430 nm
0.5700
0.5600
10% TW
0.5500
20% TW
50% TW
0.5400
0% TW + N
0.5300
10% TW + N
0.5200
20% TW + N
0.5100
50% TW + N
0.5000
0
2
4
6
Time (days)
8
10
% Change in
Absorbance at 430 nm
Tap Water and Nitrate Effects on Euglena
(% Change from Day 1 to Day 9)
15
10
5
0
-5
0% TW 10% TW 20% TW 50% TW 0% TW + 10% TW 20% TW 50% TW
N
+N
+N
+N
Concentration of Tap Water
Absorbance at 430
nm
The Effects of Tap Water Effects With and Without
Sodium Nitrate on Euglena
15
10
No Sodium Nitrate
5
Sodium Nitrate
0
-5
0% Tap 10% Tap 20% Tap 50% Tap
Water Water Water Water
Concentration of Tap Water
ANOVA
Euglena
Source of Variation
SS
df
MS
F
P-value
F crit
Sample
45.89373437
3
15.29791146
2.247482
0.108665
3.008787
Columns
153.7819531
1
153.7819531
22.59277
7.78E-05
4.259677
353.2111844
3
117.7370615
17.29726
3.39E-06
3.008787
Within
163.360525
24
6.806688542
Total
716.2473969
31
Interaction
Results and Conclusions
Chlamydomonas
• The null hypothesis that tap water will not significantly affect the
population survivorship of Chlamydomonas reinhardtii was rejected
(p<.05).
• The null hypothesis that tap water + sodium nitrate will not significantly
affect the population survivorship of Chlamydomonas reinhardtii was
rejected (p<.05).
• However, the two factor ANOVA revealed no significant interaction
between the effects of tap water and sodium nitrate (no synergistic
effects).
Euglena
• The null hypothesis that tap water will not significantly affect the
population survivorship of Euglena was rejected (p<.05).
• The null hypothesis that tap water + sodium nitrate will not
significantly affect the population survivorship of Euglena was rejected
(p<.05).
• The two factor ANOVA did reveal significant interaction between the
effects of tap water and sodium nitrate. There were synergistic effects.
Limitations and Extensions
• There were only 4 trials for each test group. This
experiment could be furthered by using larger sample
sizes.
• The cultures of algae were not sterile.
• There could be a longer duration of data collection,
which could reveal more long term effects.
• There could be more concentrations of tap water and
sodium nitrate tested.
• Only spring water was used in testing. This experiment
could be expanded by the addition of true aquatic
sources.
• There could be tests with more types of algae.
References
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“About Chlamydomonas.’’ Chlamy.org. 11 Dec. 2007.
<http://www.chlamy.org/info.html
“Algae.” Tuberose.com. 11 Dec. 2007.
<http://www.tuberose.com/Algae.html
“Chlamydomonas Reinhardtii.” Wikipedia.org. 11 Dec. 2007.
<http://www.wikipedia.org/wiki/Chlamydomonas_reinhardtii
Cloem, James E. “Eutrophication.” 12 Jan. 2008.
<http://www.eoearth.org/article/Eutrophication
Dr. John Wilson, biostatistician, University of Pittsburgh
Dr. Stanley States, Water Quality Manager for the Pittsburgh Water Authority
“Euglena.” Silicasecchidisk.conncoll.edu. 11 Dec. 2007
<http://silicasecchidisk.conncoll.edu/LucidKeys/Carolina_Key/
html/Euglena_Main.html
“Euglena.” Jracademy.com. 12 Jan. 2008.
<http://www.jracademy.com/~mlechner/archive1999/euglena.html
“Introduction to the Green Algae.” Ucmp.berkley.edu. 12 Jan.
2008.<http://ucmp.berkley.edu/greenalgae.html
“Sodium Nitrate.” Encyclopedia.com. 11 Dec. 2007.
<http://www.encyclopedia.com/doc/1E1-sodiumni.html