Bill Boehner Pittsburgh Central Catholic PJAS 2014 Grade 11

Bill Boehner Pittsburgh Central Catholic PJAS 2014 Grade 11

The technical term for saltiness in aquatic environments is halinity, from the fact that halides, or chloride, are the most abundant anions in the mix of dissolved elements Salinity is frequently reported in mg/l or ppm (parts per million)

Fertilizer runoff enters aquatic ecosystems each day. Contains high amounts of nitrogen which may cause eutrophication. From aiding in cell division to providing essential nutrients, algae can use fertilizers the same way many other plants do. Miracle-Gro

Contains the active ingredients: Urea Phosphate, Ammonium Phosphate, & Potassium Chloride. Only aids plant growth at intended concentrations. Is used often as a fertilizer in homes and at businesses.

Large diverse group of simple, and usually autotrophic, organisms Basis of the aquatic food chain Used as a bio-indicator for aquatic environments

Flagellated, unicellular, green, algae. Commonly found in fresh water and sometimes even in soil. Can grow on a simple medium of salts in the light, using photosynthesis to provide energy. Common experimental cell model.

Generally cylindrical in shape with many flagella Common algal like protist Partial heterotroph- endocytosis and photosynthesis. Nutrient-rich freshwater or in sewage systems Capable of survival in both salt and fresh water environments Common experimental model

Quantitative study of electromagnetic spectra Visible light, near-ultraviolet, and nearinfrared and the use of a spectrophotometer: can measure intensity as a function of color or light absorption Important features include the spectral bandwidth and linear range Commonly used in scientific fields such as chemistry, biochemistry, and molecular biology

To test the effects of various concentrations of salt and fertilizer on the population growth rates of algae.

Null: Miracle-Gro and Salt will not have a significant effect on the growth rate of Chlamydomonas reinhardtii and Euglena gracilis. Alternative: Miracle-Gro and Salt will have a significant negative impact on the growth rate of Chlamydomonas reinhardtii and Euglena gracilis.

Algae Chlamydomonas reinhardtii Euglena gracilis 90 Test tubes (13 x 100 mm culture tubes borosilicate) Micro-Pipettes Micro-Pipette tips Spectrophotometer (Carolina Educator) Test tube rack Miracle-Gro Sodium Chloride Soil Water Spring Water

1. Racks with tubes were set up on a table 54 cm away from a window. 2. Temperature of the room was set to a range of 21-22 Celsius. 3. The following ingredients were added to the tube to create the following experimental variable concentrations of fertilizer and salt

Control 1% Salinity 1% Fertilizer 2% Salinity 2% Fertilizer 1% Fertilizer 1% Salinity 1% Fertilizer 2% Salinity 2% Fertilizer 1% Salinity 2% Fertilizer 2% Salinity Euglena 2 ml 2mL 2mL 2mL 2mL 2mL 2mL 2mL 2mL 10% NaCl solution 0mL 0.5 ml 0 ml 1 ml 0 ml 0.5 ml 1 ml 0.5 ml 1 ml 10% Miracle Gro solution Spring Water 0mL 0 ml 0.5 ml 0 ml 1 ml 0.5 ml 0.5 ml 1 ml 1 ml 2mL 2.5mL 2.5 ml 2 ml 2 ml 2 ml 1.5 ml 1.5 ml 1 ml Soil Water 1mL 1mL 1mL 1mL 1mL 1mL 1mL 1mL 1mL Total Volume 5mL 5mL 5mL 5mL 5mL 5mL 5mL 5mL 5mL

4. The top of each tube was covered with a piece of wax paper and mixed by inversion. 5. The absorbance was taken at 430 nm using a spectrophotometer. This wavelength is within a maximum absorbance peak for chlorophyll. Thus, population density was measured indirectly by quantifying the amount of chlorophyll present in the sample. 6. Absorbance readings were taken each day for 7 days.

Absorbance Euglena Growth 0.5 Control 0.45 1% Fertilizer 0.4 1% Salinity 0.35 2% Fertilizer 0.3 2% Salinity 0.25 0.2 0.15 0.1 Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Time 1% Fertilizer 1% Salinity 2% Fertilizer 1% Salinity 1% Fertilizer 2% Salinity 2% Fertilizer 2% Salinity

Analysis of variance that compares between and within the means of the groups to determine significance Alpha=0.05 Lower p-value than alpha provides evidence to reject the null hypothesis Single factor p-value= 0.937094 Two factor p-value= 0.930769

Absorbance Chlamydomonas Growth 0.55 0.5 0.45 0.4 0.35 0.3 0.25 0.2 Control 1% Fertilizer 1% Salinity 2% Fertilizer 2% Salinity 1% Fertilizer 1% Salinity 2% Fertilizer 1% Salinity 1% Fertilizer 2% Salinity 2% Fertilizer 2% Salinity 0.15 0.1 Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Time

Single factor P-value= 0.989601 Two factor P-value= 3.54E-05 Well below the alpha value of 0.05 so this means that there was a significant effect upon the Chlamydomonas Dunnett s test was performed to see which groups were significant

Compares experimental groups back to the control group Used to determine which of the experimental groups were significant and which groups were not If T value > T-Crit value than the results were significant. T-Crit= 3.03

Chlamydomonas T value T-Crit Conclusion 1% Fertilizer 1% Salinity 1% Fertilizer 2% Salinity 2% Fertilizer 1% Salinity 2% Fertilizer 2% Salinity 0.69 3.03 Not Significant 0 3.03 Not Significant 1.38 3.03 Not Significant 3.11 3.03 Significant

Reject null hypothesis for 2% Fertilizer 2% Salinity solution with Chlamydomonas Accept null hypothesis for all other solutions Accept alternative hypothesis for 2% Fertilizer 2% Salinity solution with Chlamydomonas

Limitations Algal health and longevity might vary Mixing of salt and fertilizer with solution Extensions Use more diverse groups of algae Higher concentrations of fertilizer and salt Use more replicates

http://www.buzzle.com/articles/greenalgae-facts.html http://www.fcps.edu/islandcreekes/ecology/ euglena.html Mr. Mark Krotec, PTEI

Anova: Single Factor SUMMARY Groups Count Sum Average Variance Control 8 2.47 0.30875 0.010813 1% Fertilizer 8 2.58 0.3225 0.012879 1% Salinity 8 2.33 0.29125 0.01007 2% Fertilizer 8 2.66 0.3325 0.013564 2% Salinity 8 2.39 0.29875 0.00987 ANOVA Source of Variation SS df MS F P-value F crit Between Groups 0.009115 4 0.002279 0.19921 0.937094 2.641465 Within Groups 0.400363 35 0.011439 Total 0.409478 39

Anova: Two-Factor Without Replication SUMMARY Count Sum Average Variance Row 1 5 0.8 0.16 0 Row 2 5 0.97 0.194 0.00033 Row 3 5 1.23 0.246 3E-05 Row 4 5 1.45 0.29 5E-05 Row 5 5 1.62 0.324 8E-05 Row 6 5 1.88 0.376 3E-05 Row 7 5 2.07 0.414 8E-05 Row 8 5 2.27 0.454 0.00013 Column 1 8 2.47 0.30875 0.010813 Column 2 8 2.45 0.30625 0.01177 Column 3 8 2.46 0.3075 0.011764 Column 4 8 2.44 0.305 0.0102 Column 5 8 2.47 0.30875 0.010498 ANOVA Source of Variation SS df MS F P-value F crit Rows 0.382478 7 0.05464 539.6508 3.96E-28 2.35926 Columns 8.5E-05 4 2.12E-05 0.209877 0.930769 2.714076 Error 0.002835 28 0.000101 Total 0.385398 39

Anova: Single Factor SUMMARY Groups Count Sum Average Variance 0.14 7 2.1 0.3 0.008933 0.14 7 2.16 0.308571 0.009848 0.14 7 2.11 0.301429 0.008514 0.14 7 2.3 0.328571 0.011914 0.14 7 2.11 0.301429 0.009181 ANOVA Source of Variation SS df MS F P-value F crit Between Groups 0.004017 4 0.001004 0.103769 0.980316 2.689628 Within Groups 0.290343 30 0.009678 Total 0.29436 34

Anova: Two-Factor Without Replication SUMMARY Count Sum Average Variance Row 1 5 0.7 0.14 0 Row 2 5 0.83 0.166 0.00013 Row 3 5 1.05 0.21 1E-04 Row 4 5 1.3 0.26 0.0001 Row 5 5 1.51 0.302 0.00022 Row 6 5 1.74 0.348 7E-05 Row 7 5 1.91 0.382 7E-05 Row 8 5 2.21 0.442 0.00002 Column 1 8 2.24 0.28 0.010857 Column 2 8 2.29 0.28625 0.011227 Column 3 8 2.32 0.29 0.011943 Column 4 8 2.23 0.27875 0.011527 Column 5 8 2.17 0.27125 0.011498 ANOVA Source of Variation SS df MS F P-value F crit Rows 0.398198 7 0.056885 1367.202 9.49E-34 2.35926 Columns 0.001675 4 0.000419 10.06438 3.54E-05 2.714076 Error 0.001165 28 4.16E-05 Total 0.401038 39