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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 near-
infrared 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
0mL 0 mL 0.5 mL 0 mL 1 mL 0.5 mL 0.5 mL 1 mL 1 mL
Spring
Water 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.
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7
Ab
sorb
ance
Time
Euglena Growth
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
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
0.1
0.15
0.2
0.25
0.3
0.35
0.4
0.45
0.5
0.55
Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7
Ab
sorb
ance
Time
Chlamydomonas Growth
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
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
0.69 3.03 Not Significant
1% Fertilizer 2% Salinity
0 3.03 Not Significant
2% Fertilizer 1% Salinity
1.38 3.03 Not Significant
2% Fertilizer 2% Salinity
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/green-algae-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
