Analyzing and quantifying dissolved oxygen levels in order to determine the effects of varying levels of light exposure and

temperatures on primary productivity

Vishal Oza 02/22/11 Green Hope High School AP Biology 4th period, Mr. Jordan

Abstract The main focus of this experiment is to investigate the effects of various variables such as temperature and light exposure on the primary productivity of the corresponding ecosystem. The procedure by which the level of dissolved oxygen in the water is measured gives us a reliable look at the primary productivity of the ecosystem, and is also beneficial in determining the relationship between the levels of light exposure and primary productivity or the relationship between temperature and primary productivity. In order to test these relationships, first for the temperature, 3 sampling bottles were filled with water, each of which was at a different temperature. Then using the Winkler method the dissolved oxygen was quantified, and graphed. The results yielded that at 1oC, 20oC and 43oC there was dissolved oxygen levels (parts per million) in the water of 11.7ppm, 11.2ppm and 8.1ppm, respectively. Therefore there existed an inverse relationship between temperature and primary productivity, meaning that as temperature increased the primary productivity decreased, after that the class mean DO for all temperatures was calculated. To test light exposure, pond water was taken into 5 sampling bottles and each was covered with different layers of screens. The results for the dissolved oxygen levels for 100%, 65%, 25%, 10% and 2% light was 22.7ppm, 23ppm, 9.6ppm, and 0ppm respectively, this shows a direct relationship showing that as the amount of light increased the primary productivity increased. Later the net and gross primary productivity was calculated for both individual and class mean data in addition to initial DO, dark bottle DO and respiration rate for class mean and individual data, this data was also graphed.

Introduction

The central question surrounding this experiment is if temperature and light exposure have a significant effect on primary productivity in aquatic ecosystems. In order to determine if there is an association between the corresponding variables, the dissolved oxygen levels are measured in parts per million. The idea of aquatic primary productivity and dissolved oxygen is the topic of interest in this lab experiment; the general study of this topic starts with measuring the amount dissolved oxygen in water at various temperatures and also at various levels of light exposure, but also the study of the effect of temperature and light exposure on an aquatic ecosystem. The aquatic ecosystem consists of various species from algae and plankton to much larger aquatic marine creatures, this high

biodiversity results in an ecosystem that flourishes with life and cycling nutrients, this ecosystem productivity is usually called primary productivity, mostly measured by the dissolved oxygen present. One of the most influential organisms of the aquatic ecosystem is the algae, although one of the smallest creatures, they play a very important ecological role in cycling of nutrients and other biotic processes. In addition the algae are not only consumed as food but they also make food for themselves and nutrients that is also available and vital for the sustainability of other species. Therefore it is imperative to understand their lifecycles in order to understand their ecological impact on aquatic ecosystems, algae begin as diploid resting spores, after this stage the spore undergoes meiosis in order to make algal zoospores which are haploid, some of these zoospores consist of or make up what is called an isogamete, these isogametes fuse in something close to that of fertilization, except in that neither cell is fully embedded in the others membrane. This fusion causes the formation of the diploid zygote which in turn transforms into the resting spore also called the zygospore (Campbell, Reece, 2002). This gives us a main outline on the characteristics of the algae, and its importance to the lifestyle of other species in the aquatic ecosystem. Equilibrium in which the amount of dissolved oxygen and other gases was always almost reached but due to the random assortment of the gases this was almost never reached. The dissolved oxygen produced as bi products of metabolic reactions in cellular respiration also gives some insight on the diffusion of the gases and the time equilibrium had lasted, the time at which the dissolved oxygen concentration was constant was for approximately 1 and half seconds . Season and temperature as with primary productivity can greatly affect the rates of equilibrium and the status of gas concentration and diffusion. The Winkler Method is the method by which the level of dissolved oxygen was determined. This method shows that in order to measure the dissolved oxygen in water you must take the sampling bottle of water, add 8 drops of maganous sulfate solution and alkaline potassium iodide, later cap and mix , allow the precipitate to settle, apply one 1g of sulfamic acid powder, cap it and mix and wait for the precipitate to dissolve. Fill test tube up to the 20 mL line, and then briefly begin the titration, in which a known quantity is used to determine an unknown quantity. In order o carry out this titration you must fill the titration vial with sodium thiosulfate, apply 8 drops of starch indicator and then allow for the solution to become colorless, then read and record the dissolved oxygen levels on the titration vial, which is you unknown amount in ppm ( parts per million), (LaMotte) The Winkler

Method is extremely beneficial, due to the fact that it is very efficient, and many times more accurate than the average nomogram. Thermal pollution is a very important aspect in primary productivity as with the location of the organism in the ecosystem such as the top of a lake or the bottom of the ocean or pond, such as benthos. Thermal pollution on the other hand in general is the degradation of water quality by processes that cause a change in the overall water temperature. This can cause detrimental direct impacts on the lifestyles and ecosystems of many of the most biologically diverse marine ecosystems on the Earth, thus causing acute and long term changes in primary productivity. This is also related to the study of the light changes and its effects on primary productivity, as with any organism, without thermal energy it will die eventually, despite it being photosynthetic or not. Presence of light will ultimately enhance net (light ppm-Initial ppm) and gross primary productivity (light ppm-Dark ppm) and the levels will only increase. All of the aforementioned information and context is all relative to our hypothesis that as temperature increases primary productivity decreases and that when light level exposure increases so does the primary productivity. The scientific context presented also supports the purpose of the experiment.

Materials & Methods Pond water safety: (aprons, goggles and disposable gloves due to caustics and glass vessels) Rubber bands aluminum foil water proof markers/ tape 10 gallon pond water tank (20x12x10in) AQUA Culture 051508 aquarium air pump MK-1504 ZP AC 120V 60Hz MAX 3.5W E115699 6624 A Thermometer 2 Bulbs Sylvania Gro-Lux F40/GRO/AQ/WS/RP 40W A848 Air stones & 12 in. tubing

Dissolved Oxygen Water Quality and aquatic primary productivity Test Kit 746630A Lot No. 79190341 & Instruction Manual Code 7414/5860 (everything below is included in the kit) Source of kit and all chemical materials: Carolina Biological

Alga-Gro Concentrated Medium Item # 15-375 Fiber glass screens 50-mL Vials of Sodium Thiosulfate Alkaline Potassium Iodide Azide & lithium hydroxide Manganous Sulfate Solution Starch Indicator Sulfamic Acid Powder Spoons, 1 g Chlorella Culture Sampling Vials, 20 mL BOD Bottles and Caps titration Syringes, 1 mL Syringes, 60 cc The methods of this experiment began long before the actual formal experiment took place with the collection of the pond water from the local pond reservoir or stream, and wetland areas last August. This pond water was kept in doors in the AP Biology lab room at the back of the room on a shelf in which it was kept under 24 hrs of light until January 3rd. Later this tank was strained of the contents and drained on the 18 th; afterwards 1 Liter of the sample water was taken and diluted with 20 Liters of distilled water. After this, 6 tubes of Alga-Gro concentrate were placed in the water to set for a week. After the medium had grown biotic material in the water it was ready to test for dissolved oxygen, first the 3 bottle from the kit were filled with water so that no oxygen bubbles existed and labeled each with a different temperature, one cold, room, and hot. After this the Winkler method is implemented to determine dissolved oxygen levels, to do this take off the cap, drop 8 droplets of the aqueous manganous sulfate solution and 8 droplets of the potassium iodide azide solution. Once this is done cap and mix well by shaking up and down, and a precititate should form, allow it to settle below the shoulder of the bottle, right after this is complete add

1g spoonful of sulfamic acid powder, or its alternative 8 drops of sulfuric acid. Resume mixing by shaking up and down until the precipitate has completely dissolved, if the sample contains oxygen it should turn yellow to and orange tint. To do the titration, fill the titration vial to the 20mL line with the sample and cap, then depress the plunger of the titrator, insert titrator into the plug of the sodium thiosulfate titrating solution, then invert the bottle and slowly withdraw the plunger from the plug until it is opposite to the 0 on the scale. Turn upright and remove titrator, if not pale yellow reinsert the titrator plunger, press down and swirl gently, then remove the titrator and cap, add 8 drops of starch indicator, and it should turn blue, put the cap back on, and reinsert the titrator plunger, resume titrating until the blue color disappears and the solution becomes colorless, then record the level on the titrator barrel as dissolved oxygen in ppm (LaMotte). Record in table for each temperature, and graph this. To complete the second part take 7 water sampling bottles, and fill with the algal water from the beginning, and leave no bubbles. Mark one as I for initial and one D for dark the rest should be 100%, 65%, 25%, 10% and 2%, determine the DO for Initial immediately, then cover the D with aluminum foil and each percentage with mesh screens in the following amount, 0, 1,3,5 and 8 respectively from highest percentage to lowest. Use rubber bands to cover the bottle properly and use mesh screens on the bottoms of the bottles, and place under light. Then calculate respiration rate, class mean initial and dark DO and also gross and net primary productivity, and include graphs too. (College Board, revised 2001) Results Charts/Tables: Group # 1 2 3 4 5 6 7 8 Cold Temperature 5oC 4 oC 1 oC 0 oC 1 oC 4 oC 5 oC 1 oC Temperature and Class Dissolved oxygen (just 4th period) DO ppm 10.4 13.2 11.6 11.6 11.7 10.4 11.5 12.4 Room Temperature 26 oC 26 oC 20 oC 20 oC 20 oC 26 oC 26 oC 20 oC DO ppm 6.5 9.0 9.6 13.2 11.2 8.0 7.9 11.6 Hot Temperature 40.3 oC 36 oC 43 oC 37 oC 43 oC 41 oC 41 oC 43 oC DO ppm 6.6 8.5 8.2 8.6 8.1 7.5 9.3 9.4

%Light and Net Primary productivity (Individual Data) # of screens % light Initial DO DO Net Primary Productivity 7.2

100%

8.3

22.7

65%

8.3

23

7.35

25%

8.3

9.6

.65

10%

8.3

9.3

.5

2%

8.3

0.0

-4.15

%Light and Net Primary productivity (class average of both 1 st and 4th period) # of screens % light Initial DO DO Net Primary Productivity 4.48125

100%

9.275

18.2375

65%

9.275

17.025

3.875

3 5 8

25% 10% 2%

9.275 9.275 9.275

9.4125 5.4375 0.0

.06875 -1.91875 -4.6375

%Light and Gross Primary productivity (Individual Data) # of screens % light Dark Bottle DO Gross Primary Productivity 11.35

100%

0.0

22.7

65%

0.0

23

11.5

3 5 8

25% 10% 2%

0.0 0.0 0.0

9.6 9.3 0.0

4.8 4.65 0

%Light and Gross Primary productivity (class average of both 1 st and 4th period) # of screens % light Dark Bottle DO Gross Primary Productivity 9.11875

100%

0.0

18.2375

65%

0.0

17.025

8.5125

3 5 8

25% 10% 2%

0.0 0.0 0.0

9.4125 5.4375 0.0

4.70625 2.71875 0

Graphs:

4th period class data of dissolved O2(ppm) levels at various temperatures

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4th period class Dissolved Oxygen (ppm) data

14 12 10 8 6 4 2 0 0 5 10 15 20 25 30 35 40 45 50

Temperature

The results yielded, shows us that the relationship between temperature and dissolved oxygen in ppm is an inverse relationship, stating that the as the temperature increases the primary productivity measured by dissolved oxygen decreases. In addition to this after calculating the line of best, it becomes evident that there exists a moderate, negative, linear relationship between explanatory variable temperature and the response variable solubility of oxygen or generally the measure of primary productivity. Furthermore the error bars conclude that the residuals or simply the amount of error in a least squares regression line is at a minimum and that if plotted the residuals would show mainly a random pattern, meaning that the errors in the model are not predictable and that the corresponding model would be appropriate and reliable, as the data shows a linear pattern also. In addition a 95% confidence interval or hypothesis test would further prove that a relationship exists.

Net primary productivity at various light exposure levels

10 8 Net primary productivity (ppm) 6 4 2 0 0 -2 -4 -6 % light esposure 20 40 60 80 100 120 Net primary primary productivity for 1st and 4th period class data average Net primary productivity for individual data

The observations in this experiment showed that the data had a consistent pattern between Net primary productivity and light exposure in percents in individual and class data, this pattern can be described as direct or strong positive linear association between the variables. Meaning that as light exposure increased so did the Net primary productivity. Additionally the error bars also showed that the residuals had significantly varied among the data set and that the each of the linear models or regression lines was a reliable model in only interpolating with the data set range, due to the fact that with extrapolation any model becomes unreliable.

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Gross Primary Productivity at various levels of light exposure

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Gross Primary Productivity ( ppm)

12 10 8 6 4 2 0 0 20 40 60 80 100 120 Gross Primary productivity (Individual Data) Gross Primary productivity (class average of both 1st and 4th period)

% light exposure

With Gross Primary productivity the relationship between the percent of light exposure and gross primary productivity is similar to the relationship between net primary productivity and percent light exposure in that it is characterized by a strong or moderate positive linear relationship. Meaning that as the percent of light exposure increases the Gross primary productivity also increases. The errors in this graph are a little more significant in the vertical component, in both individual and class average data, meaning that there exists more error between gross primary product levels, meaning that the values are more varied with light exposure.

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Conclusions From thorough analysis of the data it is evident that the decision that can be made is that there does reside sufficient evidence to conclude that there is a strong correlation between temperature and dissolved oxygen and between percent light exposure and net and gross primary productivity. The hypothesis made is reliable and the data observed that supports the hypothesis is also reliable, due to the lack of significant errors. The errors or sources of mistakes we encountered was the lack of proper measuring of chemicals during the adding of the chemicals and also during the titration the vial was not being swirled as it should have been causing more sodium thiosulfate to enter and a less accurate reading for the dissolved oxygen level ppm. Also the bottle for the second experiment that had to with the percent light exposure the bottles did not have proper covering with mesh screens on the bottom and the rubber bands were not properly secured. Improvements could be to implement electronic devices that detect the dissolved oxygen amount, in order to be very accurate for good results. Set up multiple pond water samples rather than just from near Green Hope High School, for a randomization in the experiment. Extensions to this experiment could be to involve different variables such as pH effect on primary productivity, nitrate and phosphate levels and its effect on primary productivity. (Campbell, Reece, 2002)

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Bender et al.,, Michael; Grande, Karen; Johnson, Kenneth; Marra, John; Williams, Peter J. LEB.; Sieburth, John; Pilson, Michael; Langdon, Chris et al. (1987). "A Comparison of 4 Methods for Determining Planktonic Community Production". Limnology and Oceanography 32: 10851098. doi:10.4319/lo.1987.32.5.1085 Vitousek, P.M.; Ehrlich, P.R., Ehrlich, A.H. and Matson, P.A. (1986). "Human appropriation of the products of photosynthesis". BioScience (BioScience, Vol. 36, No. 6) 36 (6): 368373. doi:10.2307/1310258

In-text citations used: College Board revised, 2001 Campbell, Reece, 2002 LaMotte

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