BIOL 1209 Lab Report Cover Sheet

I certify that the writing in this assignment is my individual work and is my sole intellectual property. It does not contain the ideas, or writing of other individuals/authors.

___________________________________ Author

Matthew Landry

___________________ Date

4/24/2012

18 ____________

Lab Section #

The Effects of Interspecies Competition and Nitrite Growth and Abundance on Phytoplankton in Aquatic Systems
Introduction: Ecology is the scientific study of the interactions between organisms and the environment (Campbell and Reece 2011). Both abiotic and biotic factors can affect the interactions between organisms and their environments. Community ecology examines how interactions between species, such as predation and competition, affect community structure and organization; where as, population ecology analyzes factors that affect population size and how and why it changes through time (Campbell and Reece 2011). Two experiments were preformed one dealing with population ecology the other community ecology of a local lake. Interspecies competition in natural plant communities is highly dependent on nutrient availability (Aerts 1999). With this in mind, we conducted the first study in a laboratory setting to determine if the biotic factor of interspecies competition had an effect on population growth of Chlamydomonas due to competition between two species for resources. We created three treatments, Chlamydomonas along (control), blue green algae Oscillatoria, and a treatment with both species. Our null hypothesis for the experiment was that interspecies competition would not affect population growth of Chlamydomonas. Our alternative hypothesis was that interspecies competition would affect population growth of Chlamydomonas. Our prediction for this experiment was that in the presence of blue-green algae, Chlamydomonas would experience a decreased population growth compared to the population growth of Chlamydomonas alone. The decrease would be a result of competition between two species for resources. Nutrients are chemical elements critical to the development of plant and animal life. In healthy lakes and streams, nutrients are needed for the growth of algae that form the base of a complex food web supporting the entire aquatic ecosystem (Lindberg 2012). With this in mind, we conducted our second study based off of data collected in spring and fall of 2010 at the University Lake. In this study, we were interested in the amount of nitrite in the lake water (abiotic variable) and its effect on chlorophyll abundance (biotic variable). We were interested in determining if an increase in the amount of nitrite in University Lake would result in an increase in chlorophyll abundance due to increased nutrient availability and reduction of competition. Our null hypothesis was that the amount of nitrite would not affect the amount of chlorophyll in University Lake during spring and fall of 2010. Our alternative hypothesis was that the amount of nitrite would affect the amount of chlorophyll in University Lake during spring and fall of 2010. Our prediction for this experiment was that an increase in the amount of nitrite would result in an increased amount of chlorophyll in University Lake during spring and fall of 2010. This increase of chlorophyll would be a result of an increased availability of nutrients and reduction of competition. Methods and Materials: Population Ecology: To begin we gathered nine sterile culture tubes and label the tubes in the top third of the tube. We labeled the nine test tubes (Ch-1,Ch-2,Ch-3,

Both 1, Both 2, Both 3, BGA-1, BGA-2, and BGA-3) The Ch labeled culture tubes represented our control of Chlamydomonas alone. Culture tubes indicated with Both represented Chlamydomonas and blue-green algae together, our first treatment variable. Finally, the culture tubes labeled with BGA represented blue-green algae alone, our second treatment variable. In the three Ch and the three Both culture tubes we pipetted 1mL of Chlamydomonas culture. Next, we pipetted 1mL of the bluegreen algae (Oscillatoria) into the three BGA and the three Both culture tubes. Next, we pipetted 4mL of Min (minimal media a mixture of nitrates, phosphates, and trace metals) into the three Ch and the three BGA culture tubes. We pipetted 3mL of Min into the three Both culture tubes. We sealed all nine tubes with parafilm wax squares, and we inverted the tubes to thoroughly mix the contents. We measured the Chlamydomonas levels in each culture tube in a spectrophotometer with an initial absorbance of 750nm. We then removed the parafilm wax from the tubes and placed the tubes under fluorescent lights at room temperature. After seven, fourteen, twentyone, and twenty-eight day, we measured the Chlamydomonas levels in each culture tube using the spectrophotometer. After twenty-eight days, we used the absorbance from each test tube over the five data collections to calculate the millions of cells/mL of Chlamydomonas. We then calculated the average and standard deviation of each treatment. By graphing the results, we are able to compare the population size of each treatment over the length of the experiment. Lake Ecology: To begin we lowered a bucket in University Lake and filled it full. We then filled two collection bottles and brought these to the lab where we conducted two tests. The first test was chlorophyll determination. We began by placing a glass microfiber into a Bncher funnel using forceps. We then rinsed the filter with approximately 25mL of dH2O using gentle vacuum pressure. We then inverted one of the water samples from University Lake to mix, and then we measured 50mL of the sample in a graduated cylinder. We then filtered the measured sample through the Bncher funnel. We then picked up the filter with forceps and carefully placed it in a test tube, careful not to smash the filter or wad it into a ball. Next we added 10mL of acetone to the test tube with the filter. We placed a stopper on the tube and vortex the tube for sixty seconds. Next we placed a Whatman #1 filter into the funnel and placed the funnel in a spectrophotometer test tube. We added the entire extracted sample to the filter, and we filtered into the test tube. We then removed the filter from the funnel and discarded it. We removed the filter from the original tube with forceps and discard of the acetone waste. We blanked a spectrophotometer at 430nm using an acetone blank. We then read the sample absorbance at 430nm and recorded our data. In the second test nitrite was determined. We began by rinsing a viewing tube with collected water from University Lake. We then filed the tube to the 5mL mark with collected water sample. Next we added the contents of one NitriVer3 nitrite reagent to the sample tube. We added a stopper to the tube and inverted the contents for one minute. We incubated the tube for ten minutes. A pink color developed indicating the presence of nitrate. Next we inserted the tube into the top-right opening of a color comparator. We then placed a viewing tube with untreated sample water in the top left slot of the comparator. We rotated the disc to color match the tubes. Next we read the ppm (mg/L) nitrite nitrogen through the scale window. We recorded the results for tube 2 as mg/L nitrite nitrogen (1ppm=1mg/L). We followed these procedures for both tests in Spring 2010 and Fall 2010 and recorded our results. We then calculated the

average absorbance and standard deviation from each season for both the chlorophyll and the nitrite test. By graphing the results, we are able to compare the average chlorophyll abundance and its relation to the average nitrite abundance. Results: Population Ecology: Based on the data from the population ecology study, the control, Chlamydomonas alone had the highest growth rate with a y=0.0442x + 0.0957 The control treatment, began with the second highest population size after one week, just lower than treatment two; however, after two weeks, the population size of the control treatment exceeded that of treatment 2. The control remained with the highest growth rate for the remainder of the experiment. The second highest growth rate after twenty eight days was treatment 2, composed of both the Chlamydomonas and blue green algae with a slope of y=0.0369x +0.1627. Treatment 2 initially had the highest population size after one week; however, the increase in population size was not as significant throughout the remainder of the study. At the end of twenty-eight days, the standard deviations of the control and treatment 2 (both Chlamydomonas and blue green algae) overlapped. Finally, treatment 1, composed of blue green algae alone, had the lowest growth rate over the twenty eight days with a slope of y=0.0014x+0.0853. Treatment 1, had a very low population size throughout the experiment, never going over 0.200 million cells/mL. The treatment reached its peak at fourteen days, and then began to decrease making it different from the other two treatments, which experienced continued growth. (Figure 1). Lake Ecology: Spring 2010 had a chlorophyll abundance of 0.04. Fall 2010 had the highest chlorophyll abundance with 0.12 (Figure 2). This is a significant difference in the chlorophyll abundance between the two seasons. In the nitrite study, in spring and fall 2010 there was a standard deviation of 0.01. In spring 2010, the nitrite level in University Lake had an average of 0.01mg/mL (Figure 3). Due to the lack of 0.00mg/mL of nitrite in summer of 2010, we cannot assess the effect on chlorophyll for that season. Based on the data from chlorophyll abundance (biotic) and nitrite abundance (abiotic) in University Lake, there is no connection between these two biotic and abiotic factors. When the biotic factor increased throughout 2010, there was no correlation to the abiotic factor increasing or decreasing as well. Discussion: Population Ecology: In the population study, based on the results from the study, our null hypothesis, that interspecies competition will not affect population growth of Chlamydomonas, is supported. While the treatment containing both Chlamydomonas and blue green algae did have a slightly lower population size and rate of growth, the data was not a significant difference to conclusively draw a conclusion that interspecies completion occurred and lowered the population of Chlamydomonas. Since, both the control and treatment two had standard deviations that overlapped after twenty-eight days, our alternative hypothesis, that interspecies competition would affect the population growth of Chlamydomonas, was not supported (Figure 1). The results of this study were not the results that we had predicted. We predicted that there would be a decrease in population size as a result of competition between the two species for resources. Our results, do not agree with similar studies. The studies

conducted by Rien Aerts (1999) and L.G. Firbank (1984) produced results where treatments with two species had the lowest population size. This was a result of competition among the species for resources. Our experiment however, produced results where treatment one, with blue green algae alone, having the lowest population size. This difference could best be explained as a possible experimental flaw. To conclusively draw a conclusion of the effect of interspecies competition this study should be replicated to see if similar data results. Lake Ecology: In the lake ecology study, based on the results of the study, our null hypothesis, that the amount of nitrite in University Lake would not affect the chlorophyll abundance during spring and fall of 2010, is supported. Based on the results, we can draw a conclusion that the level of nitrite in the University Lake did not contribute to a significant increase in the amount of chlorophyll in the lake water. This result does not support our alternative hypothesis or our prediction for the study. We predicted that an increase in the amount of nitrite would increase the amount of chlorophyll in University Lake across the seasons. This increase would be a result of increased availability of nutrients and reduction of competition. Our results however, represent that the chlorophyll abundance still increased even when nitrite abundance remained constant, which was not what we expected (Figure 3). Eutrophication is the process of enrichment of lakes and streams with nutrients, and the associated biological and physical changes. Lakes and ponds are particularly vulnerable to eutrophication because the nutrients carried into them continue to build up; in contrast, to the nutrients that could be carried away in moving water (Lindberg 2012). Based on our results of our second study, we can conclude that the amount of nitrite present in the 2010 seasons has no direct link on the amount of chlorophyll abundance. Thus we can also conclude that the level of nitrite in the University Lake was not enough to cause to cause a dramatic increase in eutrophication alone. Nitrite is not the only nutrient that is present in University Lake. Other nutrients as well as other abiotic factors could have caused the increase in chlorophyll abundance or eutrophication that is present from spring to fall 2010 (Figure 2). Nutrient abundance is an abiotic variable that is density dependent and can affect the population size of an organism.

1.600 1.400 1.200 Population size (million cells/mL) 1.000 0.800 0.600 0.400 0.200 0.000 -0.200 0 5 10 15 Time (days) T1 - Blue Green Algae Linear (Control - Chlamydomonas) Linear (T2- (Both)) 20 25 30 y = 0.0014x + 0.0853 y = 0.0442x + 0.0957 y = 0.0369x + 0.1627

Control - Chlamydomonas T2- (Both) Linear (T1 - Blue Green Algae )

Figure 1: The change in population size over time when interspecies competition is present between Chlamydomonas and blue green algae.

0.16 0.14 0.12 Chlorophyll (A430) 0.1 0.08 0.06 0.04 0.02 0 Spring 2010 Time of Sampling
Figure 2: The average chlorophyll abundance in University Lake through two seasons in 2010.

Fall 2010

0.025 Nitrite (mg/mL) 0.02 0.015 0.01 0.005 0 Spring 2010 Time of Sampling Fall 2010

Figure 3: The average nitrite abundance in University Lake through two seasons in 2010.

References: Aerts, R. 1999. Interspecies competition in natural plant communities: mechanisms, trade-offs and plant-soil feedbacks. Journal of Experimental Botany, 50:29-37. Campbell NA, Reece JB. 2011. Biology. 9th ed. San Francisco (CA): Pearson Benjamin Cummings, p. 1144-1145. Firbank, L.G., Watkinson, A.R. 1985. On the analysis of competition within two-species mixtures of plants. Journal of Applied Ecology, 22:503-517. Lindberg, R. 2012, <Nutrients in Lakes and Streams, http://www.waterencyclopedia.com/Mi-Oc/Nutrients-in-Lakes-and-Streams.html> March 16, 2012. 12:36pm. CDT.