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

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

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

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

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

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

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

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

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

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

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

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 20-21 degrees Celsius. 3. The following ingredients were added to the tubes to create the following experimental concentrations of sulfuric acid.

Tube concentrations (ml) Molarity 10-2 10-3 10-4 Sulfuric Acid 10-3 Substock 10-5 10-3 Substock 0.5 0.05 0.5 0.05 0 Euglena 1 1 1 1 1 Soil Water 1 1 1 1 1 Control 0 Spring Water Total Volume 2.5 2.95 2.5 2.95 3 5 5 5 5 5

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.

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

430nm P-values: Day 1: 0.3795 Day 3: 10-13 Day 7: 10-11 Day 13: 10-22

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

T-crit= 2.689 10-2 Day 3 T-value Sulfuric Acid 10-3 10-4 10-5 12 5.1.632 1.288 Significance Significant Significant Not Sig. Not Sig. Day 7 T-value 13.748 4.208 0.705 2.114 Significance Significant Significant Not Sig. Not Sig. Day 13 T-value 29.2599 9.592 9.11 4.69 Significance Significant Significant Significant Significant

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

Limitations and Extensions! Limitations-! 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

Works cited! http://www.epa.gov/acidrain/index.html! http://www.physics.ohio-state.edu/~kagan/ phy367/p367_articles/acidrain/effects-onlakes.html! http://www.fcps.edu/islandcreekes/ecology/ euglena.htm! http://www.chemspider.com/chemical-structure. 1086.html! Mark Krotec, PTEI

Anova: Single Factor SUMMARY Groups Count Sum Average Variance 1 7 1.313 0.187571 6.7E-05 2 7 1.803 0.257571 0.000119 3 7 1.523 0.217571 0.000237 4 7 1.339 0.191286 8.36E-05 5 7 1.26 0.18 9.87E-05 ANOVA Source of Variation SS df MS F P-value F crit Between Groups 0.028157 4 0.007039 58.16652 1.05E-13 2.689628 Within Groups 0.003631 30 0.000121 Total 0.031788 34

Anova: Single Factor SUMMARY Groups Count Sum Average Variance 1 7 1.313 0.187571 6.7E-05 2 7 1.803 0.257571 0.000119 3 7 1.523 0.217571 0.000237 4 7 1.339 0.191286 8.36E-05 5 7 1.26 0.18 9.87E-05 ANOVA Source of Variation SS df MS F P-value F crit Between Groups 0.028157 4 0.007039 58.16652 1.05E-13 2.689628 Within Groups 0.003631 30 0.000121 Total 0.031788 34

Anova: Single Factor SUMMARY Groups Count Sum Average Variance 1 7 1.176 0.168 0.000108 2 7 1.739 0.248429 0.000386 3 7 1.34 0.191429 0.000301 4 7 1.107 0.158143 7.35E-05 5 7 1.057 0.151 0.000108 ANOVA Source of Variation SS df MS F P-value F crit Between Groups 0.043526 4 0.010882 55.70477 1.85E-13 2.689628 Within Groups 0.00586 30 0.000195 Total 0.049386 34

Anova: Single Factor SUMMARY Groups Count Sum Average Variance 2 7 2.093 0.299 6.97E-05 3 7 1.162 0.166 0.000938 4 7 1.049 0.149857 0.000167 5 7 0.913 0.130429 7.86E-05 0 0 #DIV/0! #DIV/0! ANOVA Source of Variation SS df MS F P-value F crit Between Groups 0.122942 4 0.030735 93.99736 6.64E-14 2.795539 Within Groups 0.007521 23 0.000327 Total 0.130462 27