Metabolism Goldish Lab Trixie

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Effects of Temperature on Metabolism Rate of Goldfish

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Trixie Le 804-644-456 Life Sciences 23l Section 1A TA: Dijana Vojnovic Effects of Temperature on Metabolism Rate of Goldfish Introduction Metabolism is defined as the catabolic and anabolic reactions that happen within a physiological system to produce energy and heat for the organism. In order to maintain a body temperature, these chemical reactions may vary and change rates. Poikilothermic organisms maintain temperatures that vary with its environment, therefore they would have a higher variance of metabolic rates. Goldfish, also known as Carassius auratus, are used as subjects to test the correlation between temperature and metabolic rates Moreover, they are tolerant of temperature fluctuations, high turbidity, low levels of oxygen, and tend to be tolerant to pollutants in comparison to other aquatic animals, making them to be highly adaptable (Nico, 2013). And according to Bowler in “Temperature Dependence of “standard Metabolic Rate,” for each 10° C increase in temperature, the metabolic rate of poikilothermic species doubles to sustain the increased temperature in the organism as well (1968). The two main substances that organisms intake for metabolic processes include oxygen and food to catalyze and sustain these metabolic reactions. As a result, the differences in oxygen consumption of goldfish will be the quantification of metabolic rate for


this experiment. The null hypothesis is that with temperature increase, there will be no difference in rate of oxygen consumption of the goldfish. However, the alternative hypothesis, supported by “Temperature Dependence of Standard Metabolic Rate,” is that higher temperature correlates to a higher metabolic rate. Materials and Method For this experiment, two goldfish will be put into an oxygen chamber with a sensor that is hooked up to a computer program called “ LoggerLite” which gives measurements of the concentration of oxygen inside the chamber at different points in time. Consequently the metabolism of goldfish will be quantified by amount of oxygen depleted in the chamber over a span of 10 minutes. However, before beginning the experiment, let the oxygen chamber and sensor sit in the water for 10 minutes to allow the sensor to adjust. In this case, the only variable that will be manipulated is the temperature of the water in order to test the hypothesis and null hypothesis. For one trial, the control group will be executed first with two fish in room temperature fish water (22 ° C). Then, the same subjects are put through the experimental group with fish water of 32 ° C After this is done, a second trial with two new subjects will be executed in the same manner. First, fill two beakers with 200 mL of fish water and weigh each beaker on the scale and record the results. Place two fish in each beaker and weigh each beaker again, and subtract from the previous weight of the beakers in order to obtain the weights of the two goldfish in each beaker. To prepare for the experimental group later, take two beakers of 300 mL fish water to incubate (warm up) while the experiment is being done.


For the control group, slowly pour one of the 200 mL beaker containing two fish into the oxygen chamber and add 200 mL of fish water to the chamber as well. At this point, there should be 400 mL of fish water inside the oxygen chamber. Before enclosing this chamber, measure the temperature of the water and record the results. Then, slowly press down on the lid until a drop of water seeps through the top hole. By this point, the lid is barely compressing the water so that only the oxygen concentrations of the water can be measured. Run the computer program, “LoggerLite-metabolism expt” and press “collect”. This program will automatically record the concentration of oxygen (mg/mL) every 20 seconds for the next 10 minutes. Copy these values into an excel sheet and create a scatterplot graph for oxygen concentration (mg/mL) and time (seconds); add a linear trend line to obtain the equation. Likewise, each dot should represent the 20 second mark and it should show the oxygen depletion over time. After 10 minutes have lapsed, start the experimental group by pouring out 300 mL of fish water from the oxygen chamber. Fill the oxygen incubator with 300 mL of warm fish water that was put into the incubator in the earlier step. There should once again be 400 mL of fish water in the oxygen chamber and use a thermometer to measure its temperature. Enclose the chamber as done for the control group, and start the computer program “LoggerLite” again. This should collect data in the same manner as it did for the control group. Once again, copy the data into excel and construct the graphs in the same manner as for the control group data. Repeat the steps done to execute the control and experimental groups to begin the second trial using the other two unused fish from the taken from the other beaker. The weight of the goldfish and the slope of the excel graphs of oxygen concentration versus time for the control and experimental group will be inputted into the UCLA LS23L Metabolism Laboratory Data, along with the data that two other student groups obtained from doing the same experiment with


two trials. In total, there would be 6 trials inputted into this database and calculate the t-test value and p-value to later determine for statistical significance of the difference of data for the controlled and experimental group. Results The data from this experiment is from Group 5 unless stated otherwise. The temperature of the control group were both 22.0 ° C and both experimental groups had the water temperature at 32.0 ° C. Figure 1-4 represent a graph of the oxygen concentration of the tank over the period of 10 minutes for the different groups and trials. The first two trials’ control groups gave similar data. The first trial control group had a slope of -0.0007 (Figure 1), the change of oxygen concentration in mg/mL over seconds, and the second trial control group had a slope of -0.006(Figure 2). However, both controls had slightly different starting concentrations of oxygen as well. Trial 1 had 5.9 mg/mL (Figure 1) whereas Trial 2 had 5.3 mg/mL of oxygen (Figure 2). Experimental group trial 1 had a slope of -0.0014, which is twice as steep as control group trial 1 (Figures 1 and 3). Experimental group trial 2 had an even steeper slope of -0.0017 (Figure 4). For both trials it was apparent that the experimental group’s goldfish had a higher rate of oxygen consumption. The weight of the goldfish in group 5 trial 1 was 10.20 grams and in group 5 trial 2 it was 9.07 grams (Figure 5). Taking into account weight of the goldfish, the LS23l database converted the slope of control group trial 1 table 5 to be -247 mg/(L*hour*kg). The slope of control group table 5 trial 2 is -238 mg/(L*hour*kg) (Figure 5). The two slopes of the experimental group trial 1 and 2 respectively became -600 and -556 mg/(L*hour*kg). There were total of 6 trials done, including the ones done by the other student groups. The other four trials’ control groups of group 6 trial 1,


group 6 trial 2, group 7 trial 1, group 7 trial 2 respectively had slopes of -425 mg/(L*hour*kg), -296 mg/(L*hour*kg), -277 mg/(L*hour*kg), -145 mg/(L*hour*kg). The other four trial’s experimental groups of group 6 trial 1, group 6 trial 2, group 7 trial 1, group 7 trial 2 respectively -850 mg/(L*hour*kg), -889 mg/(L*hour*kg), -369 mg/(L*hour*kg), -470 mg/(L*hour*kg). There was a correlation where the value of the control slope calculated by the database to be half the size of the experimental group. The mean of the controlled group slope was -247 mg/ (L*hour*kg) with a standard deviation of 92 mg/(L*hour*kg) (Figure 5). The experimental groups had a larger mean of -622 mg/(L*hour*kg). However, the experimental group had a much higher standard deviation of 207 mg/(L*hour*kg) compared to the controlled group. The t-test gave a value of 5.2770 from all the trials’ slopes and p-value of 0.002. The p-value means a 0.2% of the two groups of controlled and experimental results are not significantly different. Discussion Ultimately, the null hypothesis was rejected because the p-value that was calculated using the slopes of the controlled and experimental group of all trials was 0.002, meaning that there is only a 0.2% chance that the controlled groups and experimental groups were not statistically significant in the difference of results. And the hypothesis that fish metabolism is affected by difference in temperature is determined supported through the following results trends. As it turns out, a warmer water temperature generally meant a higher rate of oxygen consumption according. Although all four trials done by table 5 had similar oxygen concentrations in the beginning, the rate of oxygen depletion in the experimental groups ( -0.0017 mg/(mL*s) trial 1 and -0.0014 mg/(mL*s) trial 2) almost doubled their respective controlled group slopes( -0.0007 mg/(mL*s) trial 1 and 0.0006 mg/(mL*s) trial 2) (Figures 1-


4). The difference between the rates of oxygen depletion became more apparent when this parameter was then divided by the weights of the different fish (Figure 5). For example in group 6 trial 2 and group 7 trial 2 the difference between their experimental groups had almost triple the slope of their respective controlled group. Group 6 trial 2 controlled group had 296 mg/ (L*hour*kg) in comparison to its respective experimental group (-889 mg/(L*hour*kg) ). Group 7 trial 2 the control group had -145 mg/(L*hour*kg) for its slope compared to -470 mg/ (L*hour*kg). Interestingly enough, in Figures 1-4, even though the trend lines fit the overall downward trend, there seemed to be a small sinusoidal trend within the scatterplot as the oxygen concentration decreased over time. These variations may be due to levels of fish activity, which is uncontrollable. Even though the overall mean of the controlled group, -247 mg/(L*hour*kg), was significantly lower than the over mean of the experimental group, -622 mg/(L*hour*kg), the standard deviation of the two groups were high. The controlled group had a standard deviation of 92 mg/(L*hour*kg) (Figure 5) whereas the standard deviation of the experimental group was 207. The data was still not completely consistent with each other, despite dividing out the weight of the fish to normalize the values. This brings in a question to whether there may be another factor to different fish sizes that affect metabolism as well. Moreover, this experiment assumes that oxygen consumption is a direct measurement of metabolism. To strengthen the experiment, metabolism rate can be quantified to multiple different parameters rather than just one to have a more holistic understanding of metabolism. As usual, a larger subset of data would help normalize results as well.


Works Cited Bowler, K. "Temperature Dependence of “standard Metabolic Rate” in a Poikilotherm." Comparative Physiology and Biochemistry. Vol. 25. N.p.: Elsevier, 1968. 427-36. Print. Ser. 2. Harwood, John L. "Mechanisms of Temperature Adaptation in Poikilotherms." Mechanisms of Temperature Adaptation in Poikilotherms - ScienceDirect. Elsevier, 2006. Web. 18 July 2017. L, Nico G. Goldfish (Carassius Auratus) - FactSheet. USGS Nonindigenous Aquatic Species Database, 2 Aug. 2013. Web. 18 July 2017.


Tables and Figures

Figure 1: Control Group Trial 1 This graph shows how oxygen concentration, measured in mg/mL, of the chamber decreased over a span of 10 minutes (600 seconds) in the room temperature water (22.0 ° C). The metabolism rate is quantified by how steep the slope is, how much oxygen the fish consume per second. Each point represents the amount of oxygen concentration every 20 seconds. The starting amount of oxygen is defined by the y-intercept (5.9 mg/mL) and the slope (rate at which oxygen is depleted) is -0.0007.


Figure 2: Control Group Trial 2 This graph shows the data for the control group, with 22 ° C water in the second trial. It is the concentration of oxygen (mg/mL) in the chamber over the span of 600 seconds. Each point represents the 20-second mark at which the data is being recorded. The starting oxygen concentration is 5.3098 mg/mL (y-intercept) and the slope (rate of oxygen depletion) is -0.0006


Figure 3: Experimental Group Trial 1 This shows the data for the experimental group with a warmer water temperature of 32.0 ° C. Each point represents the 20 second mark, and it shows the correlation between oxygen concentration (mg/mL) and time (seconds). The slope (-0.0017) shows the rate of oxygen consumption in the chamber. And the y-intercept is representative of the starting oxygen concentration.


Figure 5: Experimental Group Trial 2 This shows the graph of the oxygen concentration (mg/mL) over time in the chamber with warm water (32.0 ° C). The time span was 600 seconds and the y-intercept (5.8) represents the oxygen concentration the chamber starts with. The slope of -0.0014 shows the rate oxygen consumption per second.

Figure 6: P-test, T-test, and Slope of All Trials Specifically, group 5 shows the results of figures 1-4 as well as the weight of the goldfish. This figure shows all the slopes (rate of oxygen depletion in tank over time) converted into hours., taking into account weight (kilograms). The control and experimental results were taken in order to determine the statistical values of the t-test, p-value, mean, and standard deviation.

Pre Lab Worksheet/Flow Chart

Metabolism Goldish Lab Trixie

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