1. The Effects of Acid Rain on Algal Growth
Alex Zapinski
Pittsburgh Central Catholic
PJAS 2015
Grade 9

2. Acid Rain Contains high amounts of nitric and sulfuric acids
Created by emissions of sulfur dioxide and nitrogen oxides.
Emissions come from both natural and man made factors
Can be deposited onto the earth in two forms: dry and wet deposition

3. Algae
Large and diverse group of eukaryotic organisms, typically autotrophs
Base of all aquatic food chains
Used as a bio-indicator for aquatic ecosystems

4. Euglena gracilis Generally cylindrical in shape with many flagella
Common algal like protist
Partial heterotroph- endocytosis and photosynthesis
Capable of surviving in both salt and freshwater ecosystems
A common lab subject

5. Spectrophotometry
Measures the amount of light that passes through a medium at a set wavelength
Commonly used in physics, chemistry, biochemistry, material science, and molecular biology

6. Past studies
Some limiting factors such as sulfuric acid can cause algal blooms
Algal blooms can lead to the deoxygenization of the pond and death of other aquatic life
High concentrations of sulfuric acid have been shown to be a hindrance to the growth of algae

7. Interaction of Components
Algae in nature is commonly affected by acid rain, which can have devastating effects on the surrounding environment
The sulfuric acid affects the growth of the euglena
The spectrophotometer is used to measure the growth of the algae

8. Purpose
To determine if sulfuric acid has significant effects on the growth of Euglena gracilis.

9. Hypothesis
Null hypothesis- the sulfuric acid will not have significant effects on the growth of Euglena gracilis
Alternative hypothesis- the sulfuric acid will have significant effects on the growth of Euglena gracilis

10. Materials Euglena gracilis
35 test tubes (13 x 100 mm culture tubes borosilicate)
Micro-pipettes
Micro-pipette tips
Spectrophotometer
Test tube racks
0.1 M sulfuric acid
Spring water
Soil water

11. Procedures
1. Racks with tubes were set up on a desk 46 cm away from a lamp. 2. The temperature of the room was set to degrees Celsius. 3. The following ingredients were added to the tubes to create the following experimental concentrations of sulfuric acid.

12. Tube concentrations (mL)
Molarity
10-2
10-3
10-4
10-3 Substock
10-5
Control
Sulfuric
Acid
0.5
0.05
Euglena 1
Soil Water
Spring
Water
2.5
2.95 3
Total
Volume 5

13. Procedures (cont.)
4. The top of each tube was covered with wax paper and mixed by inversion. 5. The absorbance was taken at 430nm using a spectrophotometer. 6. Absorbance readings were taken every two days for 14 days.

14. Anova: Single Factor
Analysis of data that compares variation within groups to variation between the groups
Alpha: 0.05
If the p-value is below the alpha, then the effects are significant

15. P-values:
Day 1:
430nm
Day 3: 10-13
Day 7: 10-11
Day 13: 10-22

16. Dunnett's Test Compares experimental groups back to the control
Determines which experimental groups produced significant variation
If T value > T-crit then the results were significant

17. 10-2 10-3 10-4 10-5 Day 3 T-value 12 5.1 .632 1.288 Significant
T-crit= 2.689
10-2
Sulfuric Acid
10-3
10-4
10-5
Day 3
T-value 12
5.1
.632
1.288
Significance
Significant
Not Sig.
Day 7
13.748
4.208
0.705
2.114
Day 13
9.592
9.11
4.69

18. Conclusion
Null hypothesis would be rejected for the concentrations of 10-2 and 10-3
Null hypothesis would be accepted for the concentrations of 10-4 and 10-5 except for day 13

19. Limitations and Extensions
Health of Euglena?
Controlled temperature
Extensions-
Use more species of algae
Use more replicates
Use more concentrations of sulfuric acid
Use concentrations of nitric acid
The synergistic effects of sulfuric acid and other chemicals

20. Works cited http://www.epa.gov/acidrain/index.html
state.edu/~kagan/phy367/P367_articles/AcidRain /effects-on-lakes.html
ena.htm
Structure.1086.html
Mark Krotec, PTEI

21. Anova: Single Factor
SUMMARY
Groups Count Sum Average Variance
E-05
E-05
E-05
ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups E
Within Groups
Total  

22. Anova: Single Factor
SUMMARY
Groups Count Sum Average Variance
E-05
E-05
E-05
ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups E
Within Groups
Total

23. Anova: Single Factor
SUMMARY
Groups Count Sum Average Variance
E-05
ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups E
Within Groups
Total

24. Anova: Single Factor
SUMMARY
Groups Count Sum Average Variance
E-05
E-05
0 0 #DIV/0! #DIV/0!
ANOVA
Source of Variation SS df MS F P-value F crit
Between Groups E
Within Groups
Total