Algal Growth in Water Sources Luke Petro CCHS ‘22

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

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

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

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.

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

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

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

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.

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

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

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.

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)

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.

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

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

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

References https://www.thoughtco.com/major-types-of-algae-373409 https://www.lenntech.com/eutrophication-water-bodies/algae.htm http://www2.canyons.edu/Faculty/robinsonc/Documents/LectureMaterials/Ch25P9Protists.ppt https://en.m.wikipedia.org/wiki/Euglena https://biologywise.com/euglena-facts https://scienceforshottaz.weebly.com/euglena.html https://www.abss.k12.nc.us/site/handlers/filedownload.ashx?moduleinstanceid=29215&dataid=26955&FileName=07.%20Euglena%20Species%20Reading%20E%20-%20global%20warming.pdf http://orbicafotografie.blogspot.com/2011/07/euglena-in-pond.html https://www.chlamycollection.org/resources/about-chlamydomonas/ https://www.ruf.rice.edu/~bioslabs/studies/invertebrates/chlamydomonas.html https://triblive.com/neighborhoods/yourallekiskivalley/yourallekiskivalleymore/8937723-74/allegheny-river-drinking

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

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

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