1. Sediment Toxic Effects on Aquatic Micro Life By Patrick Ebbert, Central Catholic, PJAS 2008

2. 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.

3. Sediment Effects on Marine Life  There are two properties of sediment that could be potentially dangerous to aquatic life. 1. 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. 2. 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.

4. 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.

5. 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.

6. 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

7. Purpose  The purpose of this experiment was to see how various concentrations of parking lot sediment affect the growth of euglena and chlamydomonas.

8. 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.

9. 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

10. Procedure 1. Arranged 48 test tubes in test tube rack; pipetted 2ml of Euglena into 24, and 2ml of Chlamydomonas into 24. 2. Pipetted sediment and spring water into each tube as displayed in chart on following slide. 3. Absorbance readings at 430nm were recorded on days 1, 4, 7, 10, and 13.

11. Procedure (cont.) Sed. A: 0% Sed. A: 10% Sed. A: 40% Sed. B: 0% Sed. B: 10% Sed. B: 40% Spring Water 3ml2.5ml1ml3ml2.5ml1ml Algae 2ml Sediment 0ml0.5ml2ml0ml0.5ml2ml Total 5ml *Repeat this table 4 times for Euglena and 4 times for Chlamydomonas

12. Algal Growth Curve

13. More Graphical Comparisons

14. ANOVA Statistical Analyses ANOVA Source of VariationSSdfMSFP-valueF crit Sample136.3308333268.165421.3441360.2857160843.55455715 Columns30.60041667130.600420.6034010.4473717024.4138734 Interaction28.01083333214.005420.2761690.7618415883.55455715 Within912.83751850.71319 Total1107.77958323 2-Factor Comparison of Sediment on Chlamydomonas

15. ANOVA Statistical Analyses (cont.) 2-Factor Comparison of Sediment on Euglena ANOVA Source of VariationSSdfMSFP-valueF crit Sample1172326.6032586163.32.7730920.0891595583.55455715 Columns110215.70671110215.70.5214220.4795126084.4138734 Interaction264199.74332132099.90.6249540.5465056653.55455715 Within3804757.02518211375.4 Total5351499.07823 2-Factor Comparison of Sediment on Euglena

16. 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.

17. 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.

18. 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

19. Daphnia 1 st -Trial Results & Conclusions  Daphnia were compared in eight total tubes, four with Sediment A, four with Sediment B.  The concentrations of sediment used were 0%, 10%, 20%, and 40%.  To the right is a graph showing the survival percentage of each tube of daphnia over a period of one week.  The Daphnia exposed to no sediment appeared to have a slower decline rate compared to the ones with 10-30% concentration.  This experiment could be expanded further to test more effects of sediment on aquatic animal groups rather than plant groups such as algae.

20. References  “Chlamydomonas reinhardtii”. Wikipedia.org.. Dec. 16, 2007.  “About Chlamydomonas”. Chlamy.org.. Jan. 7, 2008.  “Euglena”. Wikipedia.org.. Jan. 21, 2008.  “Structure of a Euglena”. Infovisual.info.. Jan. 21, 2008  Newcombe, C.P., Macdonald, D.D. “Effects of Suspended Sediments on Aquatic Ecosystems.” North American Journal of Fisheries Management.. Dec. 4, 2007.  Dr. John Wilson, Biostatistician, University of Pittsburgh.  Clare, John, PhD. “Daphnia”. Caudata.org.. Jan. 23, 2008.  “Contaminated Sediment in Water”. EPA.gov.. Dec. 4, 2007.