1. Algal Growth in Water Sources
Luke Petro CCHS ‘22

2. Purpose
To test the survivorship of Euglena gracilis and Chlamydomonas reinhardtii in the water sources of soil + spring, river, creek, and tap.

3. Rationale Water is the most valuable natural resource in the world
Much energy and cost is devoted to analyzing and purifying water
This experiment is designed to find out how two common algae survive in different types of water sources

4. Research (Euglena gracilis)
A green, single-celled, freshwater organism with a flagellum, sometimes forming a green scum on stagnant water
They usually live, in eutrophic ponds and puddles, though they are capable of surviving in fresh and saltwater.
Euglena can absorb nutrients from their environment as a survival technique, when favourable light conditions are decreasing

5. Research (Chlamydomonas reinhardtii)
A common single-celled green alga that lives in water and moist soil and typically has two flagella for swimming
Chlamydomonas is a single celled chlorophyte. Highly adaptable, these green algae live in many different environments throughout the world
Normally deriving energy from photosynthesis, with an alternative carbon source, chlamydomonas can also thrive in total darkness.

6. Research (Water Sources)
During the disinfection of tap water, Chlorine or chloramine is added to kill parasites, bacteria, viruses and germs
Creeks are characterized by slow water velocity resulting in buildup of fine, organic sediment in wetlands
Soil water is composed of mineral particles, organic matter, and air
Spring water comes from an underground source from which water naturally rises to the surface, bringing with it magnesium, calcium, sodium and potassium, which are good for algal growth
Allegheny River water has a higher level of salts due to Marcellus shale drilling waste and is slightly more polluted due to factory pollution

7. Hypotheses
Null: The varying water sources will not have a significant effect on the growth of the algae
Alternative: The varying water water sources will have a significant effect on the growth of the algae

8. Materials Euglena gracilis Chlamydomonas reinhardtii
Water samples (River, Stream, Soil and Spring, tap)
13x 100 mm Borosilicate culture tubes
Sterile 50 ml conical tube
Thermo Centra 1-5k rpm tabletop Centrifuge
Test tube rack
Micropipette
Sterile pipette tips
Carolina Student Spectrophotometer
Feit Electric 800 lumens 8.8 watts LED light bulb

9. Procedure The algae was centrifuged at 4000g for 5 minutes.
The supernatant was then removed and the pellet was placed in the variable wearers of soil + spring, river, creek, and tap at 30 ml of each sample.
5ml of the algae-water mix was then pipetted into each of the 5 culture tubes for each water and algae type. (waters were sterile filtered using a 0.2 micron syringe filter)
The culture tubes were then placed in a rack in illuminated by two Feit Electric LED light bulb for approximately 12 hours per day from approximately 6 meters away.
On December 16, 17, 18, 19, and 20, the absorbance at 430 nm was recorded.

10. P=1 P<10E-10 P<10E-10 P<10E-10 P<10E-10
7.089
*Variance
20.434
*Variance
48.375
*Variance

11. P=1 P<10E-8 P<10E-10 P<10E-10 P<10E-20
8.013
*Variance
.471
*No Variance
5.656
*Variance

12. Conclusion
In BOTH the cases of Euglena gracilis and Chlamydomonas reinhardtii, the null hypothesis, stating that the varying water sources will not have a significant effect on the growth of the algae, was rejected due to the significant variation between groups.

13. Analysis (Euglena)
As seen in the graph presenting the data for euglena, the soil and spring water (control) had the highest absorbance at the end of the five days and tap water had the lowest absorbance at the end of the five days.
Tap water could have had the lowest absorbance because the water has significant filtration and is infused with chemicals such as chlorine, making it have poor conditions for growth.
Soil + Spring could have had the highest absorbance because the water presents growth support conditions (which is why it is used for many years in algal research)

14. Analysis (Chlamydomonas)
As seen for in the graph presenting the data for chlamydomonas, the creek water had the highest absorbance at the end of the five days, and tap water had the lowest absorbance at the end of the five days.
As chlamydomonas is a very adaptive algae, the variance between the groups is lower than that of euglena.
Although this algae is very adaptive, the chemicals such as chlorine in tap water are not good for boosting growth.
Chlamydomonas could have grown the best in creek water because that is where it is commonly found, in creeks and common natural water sources.

15. Societal Impact
By chlamydomonas having an upward trend of growth, this could be significant considering that this algae is able to survive in total darkness with alternate carbon sources.
If there could perhaps be a crack in a water pipe or the algae happens to get in some other way, since this water is past the point of filtration, the algae is free to grow
This experiment could be used as a reference for looking at chemicals that are safe for humans, but prevent agal growth
Although chlamydomonas is not much of a dangerous algae, other algae with a toxic effect could be tested for this impact

16. Limitations Only using 4 water sources
The waters were not chemically analyzed
Not having the ability to fully control the light influences
Not having the ability to fully control temperature influences
Not being able to control water evaporation
Unknown health of algae samples

17. Extensions of Experiment
Analyze composition of water
Test more organisms
Present sure optimal lighting for organisms
Present sure optimal temperature for organisms
Longer light exposure time to maximize growth
Exposing algae to other forms of life in the waters

18. References https://www.thoughtco.com/major-types-of-algae-373409

19. Data (Euglena) Day 1 (December 16, 2018) Soil + Spring: .166 River:
Creek:
Tap:
Day 2 (December 17)
Soil + Spring:
.238
River:
.244
Creek:
.176
Tap:
.070
Day 3 (December 18)
Soil + Spring:
.272
River:
.250
Creek:
.180
Tap:
.048
Day 4 (December 19)
Soil + Spring
.298
River:
.266
Creek:
.202
Tap:
.068
Day 5 (December 20)
Soil + Spring:
.310
River:
.276
Creek:
.212
Tap:
.078

20. Data (Chlamydomonas) Day 1 (December 16) Soil + Spring: .174 River:
Creek:
Tap:
Day 2 (December 17)
Soil + Spring:
.150
River:
.134
Creek:
.184
Tap:
.122
Day 3 (December 18)
Soil + Spring:
.158
River:
.138
Creek:
.188
Tap:
.132
Day 4 (December 19)
Soil + Spring:
.180
River:
.162
Creek:
.206
Tap:
.144
Day 5 (December 20)
Soil + Spring:
.186
River:
.184
Creek:
.220
Tap:
.162

21. Dunnett’s Test
Equation:
t= Mi-Mc/√2MSE/nh
t-crit = 3.81