Matthew Stankey WFSC 414-500 Due: 15 November, 2013 The effect of Prymnesium parvum on the growth of other phytoplankton

species Introduction The goal of this experiment is to measure the effect of Prymnesium parvum (P. parvum), a toxic phytoplankton, on the growth of other phytoplankton species. This experiment is investigating how phytoplankton growth rates change under different salinity conditions. This experiment is also investigating how the growth rates of P. parvum change under different salinity conditions. This experiment also investigated how the growth rates of the phytoplankton change with and without the presence of P. parvum. Finally, this experiment will investigate how concentrations of P. parvum impact concentrations of chlorophyll A. We are investigating these things because it is important to understand how salinity affects P. parvum and phytoplankton, and how P. parvum affects other phytoplankton. P. parvum is the cause of common harmful algal blooms (HABs) (Johanssona & Granlib, 1999). It can cause significant damage to fish when it blooms (Giovana O. Fistarol, 2003). When P. parvum blooms damage fish in an ecosystem, the eco-tourism in the surrounding area can also be impacted. This can cause issues for not only the fish species in the area, but also for the economy of any neighboring human populations. I think that there will be a certain level of salinity that will be beneficial to the phytoplankton in the samples, but that too much will hinder their growth rates. I also think that the presence of P. parvum will negatively impact the other phytoplankton in the samples.

Matthew Stankey WFSC 414-500 Due: 15 November, 2013 Materials/Methods There were 8 treatments total, 4 with P. parvum added and 4 without P. parvum added. In each of those two subsets, there was one control, one with 1 psu (practical salinity units) salinity, one with 2 psu salinity, and one with 3 psu salinity. At t0, each student received a 125mL flask that they labeled with their treatment and name. Students with experiments that were not adding P. parvum extracted 50mL of water from Lake Somerville with a graduated cylinder and poured it into their flask. Students with experiments that were adding P. parvum extracted 45mL of water from Lake Somerville with a graduated cylinder and poured it into their flask. Every student then used a pipet with the appropriately sized pipet tip to add a set amount of nutrients (made up of nitrates, phosphates, vitamins, and trace metals). After that, students with treatments that called for salt to be added weighed salts on a gram scale and added the salt to their flasks. Students that were adding P. parvum to their flasks did so now, using a pipet to add 5mL of P. parvum to their flasks. The flasks were incubated at 25C. Throughout the experiment, the samples were 12 hours of light and then 12 hours of dark in the incubator every day. Each week, students took fluorometry readings from their samples. They did this by extracting 5mL from their flasks, injecting in into a test tube, placing the test tube into the fluorometer, writing down the result, and pouring the contents of the test tube back into the flask. Students then extracted 1mL of their sample with a pipet and inserted it into a test tube prepared with a drop of glutaraldehyde. Students then waited approximately 3 minutes, making sure that all plankton in the test tube were dead, before using a pipet to extract 10 L of the solution from the test tube for each side of the hemocytometer. Students then placed the hemocytometer into a

Matthew Stankey WFSC 414-500 Due: 15 November, 2013 microscope and counted the number of P. parvum and other phytoplankton in each of the 4 outside squares on each side of the hemocytometer. After counting, the hemocytometers were cleaned using R0 water. On the last day (t21), all students calculated the concentration of P. parvum and other phytoplankton in cells/L.

Results and Statistical Analysis The GLM statistic was run to compare concentrations of P. parvum to concentrations of chlorophyll A in all of the samples. The significant value given for the GLM statistic was 0.06. (Figure 1). The treatments with P. parvum tended to have higher fluorometry readings than the samples without P. parvum. In addition to this, the higher the salinity of the treatment, the higher the fluorometry reading. (Figure 2). A one-way ANOVA was run to compare the samples with P. parvum to the samples without P. parvum. The p-value given in the one-way ANOVA was 0.03. The P. parvum cell concentrations at t21 were: 0 for 1no; 89,250,000 for 2no; 0 for 3 no; 1,312,500 for Cno; 47,687,500 for 1Pp; 1,038,291,667 for 2Pp; 6,078,271,000 for 3Pp; 901958333.3 for CPp. The other phytoplankton cell concentrations at t21 were: 885,500,000 for 1no; 1,106,765,625 for 2no; 1,275,312,500 for 3no; 2,173,937,500 for Cno; 629,125,000 for 1Pp; 35,439,166.6 for 2Pp; 63,437,500 for 3Pp; 1,711,937,500 for CPp. (Figure 5).

Matthew Stankey WFSC 414-500 Due: 15 November, 2013 The higher the salinities of the treatments, the higher the average cell densities of P. parvum were. (Figure 3). There did not seem to be a correlation between cell densities of other phytoplankton and the salinity of the treatment from which they were taken.

Experiment Analysis and Conclusion The significant number given by the GLM statistic for fluorometry readings indicates that there is a correlation between the concentration of P. parvum and the concentration of chlorophyll A. This means that concentrations of chlorophyll A will increase if P. parvum cell densities increase. This is significant because it tells us that increased numbers of P. parvum will increase the primary productivity of a water system, but will also limit visibility and light penetration. The p-value given by the one-way ANOVA indicates that concentrations of P. parvum impacts the concentrations of other phytoplankton. This is significant because it shows that high concentrations of P. parvum decreases the species diversity of other phytoplankton in a system. This is generally bad for a system because it reduces redundancy. The graph of cell densities at t21 showed that P. parvum thrived in higher salinity conditions, but was still able to out compete other phytoplankton in 2 psu salinity conditions. It was not able to outcompete other phytoplankton in conditions with lower psu than that. This is significant because it shows us that we can use salinity to control the concentrations of P. parvum in a system.

Matthew Stankey WFSC 414-500 Due: 15 November, 2013 The average densities of P. parvum per week confirmed that P. parvum performs best in higher salinity conditions. This is important because it shows us once again that controlling the salinity of a system will help control P. parvum. The average cell densities of other phytoplankton per week showed that salinity did not significantly affect their ability to grow. This is important because it means that we can use salinity to control P. parvum without significantly impacting the ability for other phytoplankton to grow.

Matthew Stankey WFSC 414-500 Due: 15 November, 2013 Figures

Matthew Stankey WFSC 414-500 Due: 15 November, 2013 Figure 1. GLM output.

Average Fluorometry data

AVERAGE FLUOROMETRY READINGS EACH WEEK 70 60 50 40 30 20 10 0 1no 2no 3no Cno 1Pp 2Pp 3Pp CPp TREATMENTS t0 t7 t14 t21

Figure 2. Average fluorometry data for each week.

Matthew Stankey WFSC 414-500 Due: 15 November, 2013

Average cell densities per week of P. parvum

7E+09 6E+09 AVERAGE CELL DENSITY 5E+09 4E+09 3E+09 2E+09 1E+09 0 Pp t0 Pp t7 WEEK 1no 2no 3no Cno 1Pp 2Pp 3Pp CPp Pp t14 Pp t21

Figure 3. Average cell densities per week of P. parvum.

Matthew Stankey WFSC 414-500 Due: 15 November, 2013

Average cell densities per week of other phytoplankton

2.5E+09

2E+09 AVERAGE CELL DENSITY

1.5E+09

1E+09

500000000

0 Other t0 Other t7 WEEK 1no 2no 3no Cno 1Pp 2Pp 3Pp CPp Other t14 Other t21

Figure 4. Average cell densities per week of other phytoplankton

Matthew Stankey WFSC 414-500 Due: 15 November, 2013

Concentrations of P. parvum and other phytoplankton at t21

t21 t21
6078271000

CONCENTRATIONS

2173937500

1106765625

1275312500

1038291667

885500000

629125000

35439166.67

89250000

1NO

2NO

3NO

1312500

CNO

47687500

1PP

2PP

3PP

63437500

TREATMENTS

Figure 5. Concentrations of P. parvum and other phytoplankton at t21.

Works Cited Giovana O. Fistarol, C. L. (2003). Allelopathic effect of Prymnesium parvum on a. MARINE ECOLOGY PROGRESS SERIES, 255, 115-125. Johanssona, N., & Granlib, E. (1999). Influence of different nutrient conditions on cell density, chemical composition and toxicity of Prymnesium parvum (Haptophyta) in semicontinuous cultures. Journal of Experimental Marine Biology and Ecology, 239(2), 243258.

901958333.3

CPP

1711937500

Matthew Stankey WFSC 414-500 Due: 15 November, 2013