The Effect of Iron Fertilization Concentrations on Aquatic Ecosystems

Sierra Turpin Rockdale Magnet School for Science and Technology 930 Rowland Road Conyers, GA 30012 May 9, 2013

Table of Contents

Inroduction______________________________________________________________3 Literature Review_________________________________________________________5 Methology_______________________________________________________________9 Data Intrepretation_______________________________________________________12 Discussion and Conclusions _______________________________________________20 Acknowlegments________________________________________________________22 Literature Cited_________________________________________________________23 Appendixes____________________________________________________________25

Introduction: A major problem that we face today is ocean acidification which causes different effects within marine organisms. Ocean acidification is the process of carbon dioxide being absorbed by seawaters causing a decrease in PH levels and ion concentrations. A method that is used to reverse this process is iron fertilization which allows for an exponential growth in phytoplankton blooms. With iron fertilization the process of carbon sinking is made possible. When the iron concentrations are deposited into the ocean it stimulates the phytoplanktons growth and this allows for the phytoplankton to absorb more carbon dioxide from the ocean. The phytoplankton will also take in any excess carbon that is left in the atmosphere though photosynthesis. Our main goal is to observe and discover if iron fertilization poses any possible sign of threat to marine organisms. If iron fertilization exhibits negative reactions to the ghost shrimp and phytoplankton, the organism in which we are testing on, than we could possibly reduce the environmental damage inflicted by humans. Phytoplankton will be used in the experiment because a main result of iron fertilization is its ability to enhance the growth of the phytoplankton. With the phytoplankton increased size it allows for more carbon to be absorbed. This will be a great sample in order to test the effect of iron fertilization concentrations on marine ecosystems, due to the fact of their ability to absorb iron fertilizers. Ghost shrimp will also be present in the experiment to see the effects of the fertilizers on their health as well as the effects the phytoplankton will have on them. There are two major hypotheses in this experiment. If iron oxide is tested in water, then it will increase phytoplankton growth, and reduce oxygen levels in the water; and if low, medium, and high concentrations of phytoplankton and iron oxides are tested, then high concentrations of phytoplankton and iron oxide will result in the greatest decrease in ghost shrimp population.

These hypothesizes were made because heavy metals tend to have a negative effect on marine ecosystems. The metals become toxic and tend to weaken the immune system leaving organisms open to disease. The metals also leach chemicals into the water causing an increase in pH levels, and the toxicity results in the production of oxidizing agents said to be harmful to the tissues of living organism. (Cambridge Univ, 2008) Since iron is a metal, the assumption that is was harmful to the marine life and caused negative effects was made.

Literature Review: Oceans absorb carbon dioxide though the atmospheric carbon of phytoplankton through photosynthesis. Phytoplankton is responsible for half of the carbon fixation on Earth. (Mark, 2003) It occurs by the carbon being drawn into the body of plants during photosynthesis. When phytoplankton die they carry the stored carbon with them. Thus this may have an effect on the marine life that dwells at the bottom of the sea floor. The waste that sinks is also known as biological or soft tissue pump. This research will help us when we conduct our experiment because we will be able to understand if there is a possible connection of the iron oxide and the growth of the phytoplankton, and why the growth would occur. (Dean, 2009) The Research of Deep Carbon Export from a Southern Ocean Iron-Fertilized Diatom Bloom was observed by V. Smetacek and his colleagues. The experiment was tested over a 5 week time period and was created in the Antarctic Circumpolar Current (Smetacek 2012) region of ocean. What the researchers were trying to test was the effectiveness of iron fertilization in sequestering carbon. There were so problems while testing data because it was tested in the ocean, they were still managed to collect the appropriate data. The diatom bloom did not appear until the fourth week and starting in the fifth week. A large part of the bloom had died(Smetacek 2012). This research will help us in discovering how much iron fertilizer we should use in our experiment. Smetacek and his colleagues had used approximately 7 tons of iron sulphate dissolving into 54m3, in which would go into of ocean (Smetacek 2012). This would be a new way of developing a theory that is related to atmospheric Carbon dioxide. Over time some people looked into how iron made the phytoplankton grow. They would use phytoplankton stalks, and found out that when iron is added then dramatic blooms will occur

within the phytoplankton populations. Since the phytoplankton blooms were increasing there were also increases in the waste of zooplankton. The zooplanktons are what feed on the phytoplankton thus their extra waste contribute to the fertilization problems that are already occurring. (Landry, 2000) Natural ocean fertilization can come from the leaching of iron from volcanic islands. The addition of iron fertilization can increase primary productivity. It can also enhance carbon exports in the ocean. High concentrations of nutrients and nitrates and even phosphate allow for large blooms in the spring for phytoplankton. (Billet, 2007) The ocean is a natural reservoir of carbon and it is one of the largest. It plays a major role in the regulation of atmospheric carbon dioxide, and also climate because it is due to large heat capacity. Oceans collect about one-third of the emissions from burning fossil fuels. With the increased atmospheric CO2 concentrations =, global temperatures increase in the environment. (Martin, 2010) Iron fertilization will lead to iron enrichment. Global warming has caused many countries to have experienced a 2 degree Celsius temperature increase. Natural carbon will sink to reduce atmospheric carbon concentration. (Gussow, 2009) Iron fertilization is to help enhance the oceanic carbon sink in marine ecosystems, but by trying to reduce CO2 levels in the water by using iron cause dangerous affects to marine organism. For example heavy metals will and cam leach chemicals in the water, these chemicals cause in increase in pH and organism that are not adapted to those kind of high changes in pH will die and therefore cause an overall decrease in diversity.

Since we were happening to use ghost shrimp for our project more detail about their growth, reproduction, and survival was observed and discussed. The reason ghost shrimp are called ghost shrimp is because they are very transparent and hard to notice. In fact when they eat you can somewhat see the food in their stomach. Female ghost shrimp grow to about 1 and a half inches while male ghost shrimp grow up to approximately one inch. Ghost shrimp can be very aggressive towards each other and therefore there should be a moderate amount of ghost shrimp in one tank. A reasonable number would be one shrimp for every one gallon or 4 litters of water. Since ghost shrimp are scavengers they will eat just about anything they can find. A good choice of food for them would be to give them flakes, but if they seem to grow tiresome of it you can add small frozen food chunks. (Shandilya, 2010) Previous studies of iron fertilization have shown that iron fertilizations cause increase in phytoplankton growth, and how carbon which is primarily in the form of the bicarbonate ion and sulfur as sulfate, are plentiful throughout the water and cause for an increase in plant nutrients. These nutrients from the plant help aid in the growth of phytoplankton. Nitrate, phosphate, and silicate are major contributors for the biomass in the sea. (Coale) The reason for using ghost shrimp in this project was because there was a need for another organism to be involved with the phytoplankton and the iron fertilizers. Since ghost shrimp happen to be bottom feeders they would be a good organism to test because when the phytoplankton dies they will leave the carbon in their body and thus when they sink the carbon will go down with them. This is a process of carbon sinking which is an effect of iron fertilization.

This is why interested studies were of how the shrimp would react to respond to the extra carbon at the bottom or if the fertilizers would cause in increase in their growth as well. Since ghost shrimp will eat algae the iron fertilizers contributed major increases of algae blooms. The problem now is how ocean acidification causes increased amounts of the pH levels and ion concentrations. Iron fertilizers are used to help reduce the effects of excess carbon but heavy metals prevent marine organisms with a dangerous environment to live in.

Methodology: Safety Discussion: When conducting this experiment please make sure that you are wearing goggles and gloves at all time when handling the iron oxide. You may need to wear a mask when opening the iron oxide or even open it under a fume hood to avoid inhalation. Also while working make sure that you have supervision from an adult or a teacher and that they have knowledge of where you are and what you are doing. When your experiment is finished you can dispose of the excess water in containers that your teacher or adult can supply you. Materials: Phytoplankton (fresh water) Iron Oxide Spec 20 Venier Probes ( Dissolved Oxygen, gas Probe; pH sensor) Water Tank Ghost Shrimp Fish Food Sand Dinoscope Pipettes Auto Clave CaCO3 Soil

Filter Light Centrifuge Beaker Test Tubes Cuvette

Procedure Summary: First make sure that you have ordered all the above materials for the experiment. Now that you have yours materials you want to prepare three tanks in order to do the first part of the experiment. To prepare the tanks add sand, and about three gallons of water. In the first tank you will have just water and phytoplankton (Control). In the second tank you will have water and iron oxide (Control 2), and in the third tank you want to put both phytoplankton and oxide in with the water. Make sure that you use soil water for the phytoplankton in order to help their development. During the testing be sure to measure the PH, absorbance, and dissolved oxygen levels. For the second part of the experiment you need to first prepare five tanks. To prepare the tanks you will need to add sand, and then the tap water which has had the chlorine removed because the ghost shrimp are fresh water and will die if the water is not cleaned. Once you have your tanks prepared you can now start. In the first tank you will add only water and ghost shrimp (Control). In the second tank you will add only phytoplankton and water (Control).For the rest of the remaining tanks you will need to add three different concentrations of iron oxide into each tank, but first add ghost shrimp and phytoplankton in all the tanks. In the first out a high

concentration, the second one should have a medium concentration and the third once will have a high concentration. You will test a before and after so in about 5 days be sure measure ghost shrimp health. In order to measure the health you have to place the ghost shrimp into a test tube with just a small amount of water. Once you have them in the tube you can see their heart with a dinoscope which will enable you to count the heart rate.

Data Interpretation: For our experiment we tested the effect of iron oxide on phytoplankton growth and the effect of iron oxide on the health of ghost shrimp. Our hypothesis was that the iron oxide would have a negative impact on the shrimps heart rate and cause an increase production of phytoplankton. We also made a hypothesis that the oxygen concentrations would be reduced with the addition of the iron oxide. In order to interpret the raw data and figure out if our hypothesis would be supported or not we ran our testing with ANOVA. In ANOVA we did a two way test because we had multiple IVs, which were the phytoplankton and the iron oxide. We ran three separate tests for our part one of the experiment. The first test that we ran was the effect of iron oxide and phytoplankton on oxygen concentrations. Our hypothesis was not supported because the P-value for that test was about .05 at .54. For the next test it was significant that the iron oxide and ghost shrimp did have an effect on the PH levels. The pvalue for this was .012 which is less than .05. For a large majority of the days it seemed as if the PH levels were higher in the tanks that had iron oxide, but also they were about neutral PH in the tank that had only water. This tells you that the iron oxide have an effect on the PH levels. The finally test for part one of the experiment resulted in the iron oxide not having an effect on the growth of phytoplankton. The P-value for the test was actually pretty high and was .464, and to be significant than the p-value should have been at .05 or less. On the second part of the experiment we found out that the iron oxide did have an effect on the health of the ghost shrimp. During the ANOVA test it had a .002 p-value with is way below .05.

Average PH Levels Per Tank Average PH Treatments Level 1 Phytoplankton 6.52 2 Phytoplankton 9.49 3 Phytoplankton 8.68 4 Phytoplankton 7.14 5 Phytoplankton 7.68 1 Iron Oxide 6.35 2 Iron Oxide 8.72 3 Iron Oxide 8.71 4 Iron Oxide 6.98 5 Iron Oxide 7.26 1 Iron Oxide + Phytoplankton 6.34 2 Iron Oxide + Phytoplankton 8.47 3 Iron Oxide + Phytoplankton 8.5 4 Iron Oxide + Phytoplankton 6.66 5 Iron Oxide + Phytoplankton 6.71 Table: 1 This table represents the different types of treatments and their effects on the PH level for each Day tank Average Oxygen Levels Per Tank Day Treatments Average Oxygen Level 1 Phytoplankton 5.6 2 Phytoplankton 7.7 3 Phytoplankton 7.3 4 Phytoplankton 7 5 Phytoplankton 7 1 Iron Oxide 6.1 2 Iron Oxide 6.8 3 Iron Oxide 6.3 4 Iron Oxide 6.4 5 Iron Oxide 6.5 1 Iron Oxide + Phytoplankton 5.8 2 Iron Oxide + Phytoplankton 7 3 Iron Oxide + Phytoplankton 6.2 4 Iron Oxide + Phytoplankton 6.6 5 Iron Oxide + Phytoplankton 6.2 Table: 2 This table represents the different types of treatments and their effects on the oxygen level for each tank

Avergae PH Levels Per Tank Column1 Column2 Day Treatments Average Oxygen Level 1 Phytoplankton 1.003 2 Phytoplankton 0.0037 3 Phytoplankton -0.0003 4 Phytoplankton 0.012 5 Phytoplankton -0.004 1 Iron Oxide 1.003 2 Iron Oxide 0.012 3 Iron Oxide -0.002 4 Iron Oxide 0.014 5 Iron Oxide 0.0017 1 Iron Oxide + Phytoplankton 1.002 2 Iron Oxide + Phytoplankton -0.002 3 Iron Oxide + Phytoplankton -0.002 4 Iron Oxide + Phytoplankton 0.013 5 Iron Oxide + Phytoplankton 0.005 Table 3: This table represents the different types of treatments and their effects on the absorbance level for each tank

Average Heart Rate Ghost Shrimp Heart Day Treatment Rate Final Phytoplankton 82 Final Iron Oxide 121 Final Low Concentration 123 Final Medium Concentration 115 Final High Concentration 119 Initial Phytoplankton 99 Initial Iron Oxide 128 Initial Low Concentration 131.7 Initial Medium Concentration 134.5 Initial High Concentration 133 Table: 4 This table shows the before and after of the the ghost shrimp heart rate. From the table you can see that the heart rate increased when the iron oxide was added

Two-way ANOVA: Average Oxygen Concentration versus Day, Treatment

Source Day Treatment DF 4 2 P 0.014 0.054

From the data shown you can see the p-value, which is the highlighted part, was under .05 which created the conclusion that oxygen concentrations are not affected by iron oxide or phytoplankton. The highlighted number is all the below data tables represent the p-value which must be under .05 to be significant.
Two-way ANOVA: Average Ph level versus Days, Treatments
Source Days Treatments DF 4 2 P 0.012

Two-way ANOVA: Average Absorbance Level versus Date, Type of Treatment

Source Date Type of Treatment DF 4 2 P 0.464

Two-way ANOVA: Ghost Shrimp Heart Rate versus Dates, Type of Treatments
Source Dates Type of Treatment DF 1 4 P 0.002

Using the Descriptive data if was observed that Iron oxide only had an effect on the PH levels and the health of ghost shrimp, but had no effect on the phytoplankton growth and oxygen concentrations.

Figure: 1 This graph represents the different types of treatments and their effects on the PH level for each tank

Figure: 2 This table shows the before and after of the the ghost shrimp heart rate. From the table you can see that the heart rate increased when the iron oxide was added

Figure 3: Table: 2 This graph represents the different types of treatments and their effects on the absorbance level for each tank

Figure: 4 This table represents the different types of treatments and their effects on the oxygen level for each tank

Figure 5: The beakers where the phytoplankton was placed on order to grow

Figure 6: The ghost shrimp when they are in the tanks

Discussion and Conclusion: From our hypothesis was that the iron oxide would have a negative impact on the shrimps heart rate and cause an increase production of phytoplankton. We also made a hypothesis that the oxygen concentrations would be reduced with the addition of the iron oxide. In ANOVA we did a two way test because we had multiple IVs, which were the phytoplankton and the iron oxide. The first test that we ran was the effect of iron oxide and phytoplankton on oxygen concentrations. Our hypothesis was not supported because the Pvalue for that test was about .05 at .54. For the next test it was significant that the iron oxide and ghost shrimp did have an effect on the PH levels. The p-value for this was .012 which is less than .05. For a large majority of the days it seemed as if the PH levels were higher in the tanks that had iron oxide, but also they were about neutral PH in the tank that had only water. This tells you that the iron oxide have an effect on the PH levels. The finally test for part one of the experiment resulted in the iron oxide not having an effect on the growth of phytoplankton. The P-value for the test was actually pretty high and was .464, and to be significant than the p-value should have been at .05 or less. On the second part of the experiment we found out that the iron oxide did have an effect on the health of the ghost shrimp. During the ANOVA test it had a .002 p-value with is way below .05. Overall only the PH and ghost shrimp health had significant data and proved to support our hypotheses. There happened to be many sources of effort in our experiment. One of them was that we had to continuously buy new ghost shrimp because the ones that came in kept dying before we could start treatments on them. Also the ghost shrimp would jump out of the tank which

could have some impact on their health. The water from the tanks also evaporated so we had to fill them back up to the appropriate location. By doing this we may have ruined the amount of de chlorine solution that we had applied to the tanks initially. For future resource we would try to limit the different IVs and focus on one variable in order to have better data.

Acknowledgments: Ariel Elliot: She helped me in getting my ghost shrimp into the test tubes Vinh Tinh: He helped me in getting the ghost shrimp into the test tubes Arianna Gibbs: She helped in catching the shrimp and putting them in the beaker for me to then place in the test tube. John Hendrix: Helped me obtain my materials that I needed and helped with my research processes Scott Bolen: Made it possible for my research plan to be approved and helped me though my research plans with ideas. Mary Mansell: Showed me how to use the autoclave Jennifer Marshall: Helped with using the Spec 20

Literature Cited: http://www.aquaticcommunity.com. (2008). Retrieved from http://www.aquaticcommunity.com/inv/ghostshrimp.php Hugh Powell (2007, November 13). www.whoi.edu. Retrieved from https://www.whoi.edu/oceanus/viewArticle.do?id=34167 Powell, H. (2008). Oceanus. In Woods Hole Oceanographic Institute. Retrieved from http://www.whoi.edu/oceanus/viewArticle.do?id=35609ionid=1000 Dean, J. (2009). Iron fertilization: a scientific review with international policy recommendations. Retrieved from http://environs.law.ucdavis.edu/issues/32/2/dean.pdf Abate, R., & Greenlee, A. (2010). Sowing seeds uncertain:ocean iron fertilization, climate change, and the international environmental law framework. Retrieved from http://law.famu.edu/download/file/Abate & Greenlee - Ocean Iron Fertilization Article Final PDF.pdf Landry, M.(2000). Biological Response to Iron Fertilization in the Eastern. (vol.201: 27-42). Marine Ecology Progress. Retrived from http://www.int-

ves.com/articles/meps/201/m21p027.pdf Dean, Jennie. (2009). Iron Fertilization a Scientific Review with International Policy Recommendations. Retrieved from envions.law.ucdavis.edu/issues/32/2/dean.pdf Tamaki, A. (1997). Life History of the Ghost Shrimp, callanassa Japonica. Retrived from http://decapoda.nhm.org/pdfs/23291/23291.pdf

Michael, J. (2005). Performed Phosphate, Soft Tissue Pump and Atmospheric CO2. Retrived from http://puddle.mit.edu/~miak/papers/ItoFollows-performed-JMR2005.pdf

Appendices: Detailed Procedures: Safety Discussion: When conducting this experiment please make sure that you are wearing goggles and gloves at all time when handling the iron oxide. You may need to wear a mask when opening the iron oxide or even open it under a fume hood to avoid inhalation. Also while working be sure that you have supervision from an adult or a teacher and that they have knowledge of where you are and what you are doing. When your experiment is finished you can dispose of the excess water in containers that your teacher or adult can supply you. Materials: Phytoplankton (fresh water) Iron Oxide Spec 20 Venier Probes ( Dissolved Oxygen, gas Probe; pH sensor) Water Tank Ghost Shrimp Fish Food Sand Dinoscope Pipettes Auto Clave CaCO3

Soil Filter Light Centrifuge Beaker Test Tubes Cuvette

*note: Please feed the ghost shrimp at least once a day with the fish food, about four pinches of food should be added for one tank. *note: Please use algae-grow to help the development of the phytoplankton

Part One: The effect of iron oxides and phytoplankton on water clarity and oxygen levels 1. Require all materials that are needed from above before the start of the experiment 2. Set up 3 tanks for the iron oxide and the phytoplankton to be placed. To prepare the tanks add three gallons of water to each. When preparing water use soil-water. To prepare the soil-water be sure to only use potting soil and that the soil does not have peat or sphagnum. *Also do not use any commercial Fertilizers. 3. Be sure that you filter out the soil water so that there is not such a great amount of soil left. If there was to much soil then you would not be able to see the growth of the phytoplankton 4. Once you have the soil, fill the vessel with 3/4 of distilled water and steam for two hours and for two consecutive days using an auto clave.

5. You need to also clean your sand and you can do this by using the autoclave or by using an oven. 6. Fill the tanks up with regular tap water but be sure to de-chlorinate the water because since the phytoplankton you will need should be ones that live in fresh water so they must be kept in fresh water. 7. Add approximately 1 milliliter of the de chlorinator for every gallon of tap water you add to the tank 8. In the first tank add only the phytoplankton to the water. This will be the control to see the changes in oxygen levels. Put an adequate about of phytoplankton in. 9. In the second tank add only iron oxide to the water for this will be the second control. Put a medium concentration of iron oxide in the tank. 10. For the third tank you can now add both phytoplankton and iron oxide to the water. Put the same amount of phytoplankton as the first tank and a medium concentration of iron oxide. 11. After watching and observing the tanks for 5 days keep record everything that has occurred. 12. Be sure to record the dissolved oxygen, ph, and absorbance levels every day for 5 days. You can then see how phytoplankton and iron oxide would affect the oxygen levels in a marine ecosystem. Fill in your table with the data you obtained.

Part Two: The Effect of Iron Fertilization Concentrations on Ghost Shrimp

1. First you will need to set up five tanks to place the phytoplankton, ghost shrimp, and iron oxide. In all five tanks put the sand into the tanks first. 2. You will then put tap water that has be de chlorinated into the tanks and you will add one gallon for every one ghost shrimp. Since you will be using three ghost shrimp per tank than you will need three gallons of water. 3. Before you add the water be sure to keep it between 18 C to 25 C. Once the water is added in the first tank you will then add three ghost shrimp this, will be your first control. You will then observe and see how the ghost shrimp are affected. 4. In the second tank, which will be your second control, you will add only phytoplankton and see how their growth and development is affected. 5. Now that you have your controls set you will then add three different concentrations of iron oxide to the remaining three tanks. In the first tank put a low concentration, in the second tank put a medium concentration and in the third tank you will add a high concentration. 6. You will then add three ghost shrimp to each tank and an adequate amount of phytoplankton to each also. 7. You are trying to see how the different levels of iron oxide are affecting the health of ghost shrimp and how it affects the growth of the phytoplankton. 8. After you test for about 5 you can now measure ghost shrimp health observance under microscope, measure heart rate using microscope, health scale, and if they had died is required. To measure the phytoplankton growth you will need a spec 20.

Figure 9: Experimental setup of the tanks

Flow Chart:

Qualitative Scale: Check and record ghost shrimp heart rate or beats per minute (BPM) for each ghost shrimp Take an initial reading before adding the ghost shrimp into the tanks to establish a baseline for the heart rate. That reading will give you the average rate (BPM) for a normal healthy ghost shrimp. Each day record the heart rate of the ghost shrimp. If you notice that the BPM is lower than the initial value the temperature of the water most likely increased, and if the temperature is higher the heart rate will increase. This happens because of a higher metabolic activity and more chemical reactions occurring within the body. If you notice your ghost shrimp have died within a couple days or even hours record which shrimp from which tank died to see which concentration of iron oxide had the greatest affect. Ghost shrimp should stay a clear color if you note any other color changes occur, record and draw observations.

Raw Data: Oxygen Concentrations Per Tank Per Trial (mg/l) Day Tank 1 Tank 1 Tank 1 Tank 2 Trial 1 Trial 2 Trial 3 Trial 1 1 5.8 5.0 5.9 6.1 2 8.0 7.8 7.4 6.9 3 7.4 7.7 6.8 6.5 4 7.6 7.3 6.1 6.8 5 7.8 7.2 6.1 6.3

Tank 2 Trial 2 5.7 6.8 6.0 6.3 5.8

Tank 2 Trial 3 6.5 6.6 6.3 6.2 7.4

Tank 3 Trial 1 5.3 6.7 6.2 7.6 6.2

Tank 3 Trial 2 5.9 7.5 6.1 5.3 6.3

Tank 3 Trial 3 6.2 6.9 6.3 6.2 6.1

pH Levels Per Tank Per Trial Day Tank 1 Tank 1 Trial 1 Trial 2 1 6.32 6.67 2 9.52 9.7 3 8.44 8.77 4 7.19 7.12 5 8.00 7.60

Tank 1 Trial 3 6.56 9.23 8.84 7.13 7.44

Tank 2 Trial 1 6.41 8.96 8.77 7.08 7.31

Tank 2 Trial 2 6.33 8.77 8.73 7.00 7.26

Tank 2 Trial 3 6.30 8.44 8.62 6.87 7.22

Tank 3 Trial 1 6.31 8.44 8.56 6.73 7.09

Tank 3 Trial 2 6.32 8.51 8.50 6.65 6.96

Tank 3 Trial 3 6.38 8.48 8.45 6.59 6.08

EDD: Part 1: Title: The effect of iron oxides and phytoplankton on water clarity and oxygen levels Hypothesis: If iron oxide is tested in water, then it will increase phytoplankton growth, and reduce oxygen levels in the water. Independent Variables Phytoplankton in water Control 1 3 Trials Iron Oxide in Water Control 2 3 Trials Phytoplankton and Iron Oxides in Water 3 Trials

Dependent Variables: absorbance (tested with spec 20) and oxygen levels (tested by veneer probe) Constants: amount of water, length of time being tested, amount of iron oxide, temperature, amount of light

Title: The Effect of Iron Fertilization Concentrations on Ghost Shrimp Hypothesis: If low, medium, and high concentrations of phytoplankton and iron oxides are tested, then high concentrations of phytoplankton and iron oxide will result in the greatest decrease in ghost shrimp population. Independent Variables Ghost shrimp and phytoplankton with water Control 1 3 Trials 3 Trials Ghost shrimp in water Control 2 Phytoplankton and iron oxide in water low concentration 3 Trials Phytoplankton and iron oxide in water medium concentration 3 Trials Phytoplankton and iron oxide in water high concentration 3 Trials

Dependent Variables: Ghost Shrimp Health (Observance under microscope, measure heart rate using microscope, health scale, and if they had died) Constants: amount of water in the tank, tank size, amount of light, type of shrimp, water temperature, amount of shrimp in each tank, food.