1. Tap Water Intrusion Effects on Algal Populations
Anthony DeRenzo
Grade 9
Pittsburgh Central Catholic High School

2. 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.

3. Types of Surface Runoff
Petroleum
Pesticides
Many organic and inorganic chemicals
Silt
Fertilizers
Tap water

4. 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.

5. Common Components in Tap Water
Calcium
9.84 mg/L
Iron
mg/L
Potassium
1.36 mg/L
Magnesium
1.72 mg/L
Manganese
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

6. 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

7. Eutrophication
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.

8. 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.

9. 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.

10. 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.

11. 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.

12. 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.

13. Purpose
Test tap water and a mixture of tap water + sodium nitrate on two types of algae: Chlamydomonas reinhardtii and Euglena sp.

14. 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.

15. Materials 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

16. 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.

17. Procedure 2 Algae Tap Total 2 ml Spring Water 3 ml 2.5 ml .5 ml
Sodium
Nitrate
.05 ml [.1%]
Total
5 ml

18. 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.

22. ANOVA Chlamy. 0.000243 Interaction 10.68706 3.562354 0.351002 0.788772
Source of Variation SS df MS F
P-value
F crit
Sample 3
1.7E-05
Columns 1
Interaction
Within 24
Total 31

26. ANOVA Euglena 7.78E-05 Interaction 353.2111844 117.7370615 17.29726
Source of Variation SS df MS F
P-value
F crit
Sample 3
Columns 1
7.78E-05
Interaction
3.39E-06
Within 24
Total 31

27. 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.

28. 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.

29. References “About Chlamydomonas.’’ Chlamy.org. 11 Dec. 2007.
<
“Algae.” Tuberose.com. 11 Dec
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“Chlamydomonas Reinhardtii.” Wikipedia.org. 11 Dec
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Cloem, James E. “Eutrophication.” 12 Jan
<
Dr. John Wilson, biostatistician, University of Pittsburgh
Dr. Stanley States, Water Quality Manager for the Pittsburgh Water Authority
“Euglena.” Silicasecchidisk.conncoll.edu. 11 Dec < html/Euglena_Main.html
“Euglena.” Jracademy.com. 12 Jan
<
“Introduction to the Green Algae.” Ucmp.berkley.edu. 12 Jan <
“Sodium Nitrate.” Encyclopedia.com. 11 Dec
<