Metabolic Rates Using Rates of Carbon Dioxide Produced and Oxygen Consumed in Terrestrial Animals

Metabolic Rates Using Rates of Carbon Dioxide Produced and Oxygen Consumed in Terrestrial Animals Group 3 Amores, Caballes, Dizon, Ocampo, Whang

Outline I. Introduction II. Materials and Methods III.Results IV.Discussion V. Summary

Introduction Metabolism - Greek word metabolē (change) - Totality of chemical reaction in the body to sustain life

(Reece et al., 2010)

Introduction Anabolism vs. Catabolism - Anabolism: consumption of energy to make complex molecules from simple molecules - Catabolism: release of energy from breakdown of complex molecules to simple molecules

(Reece et al., 2010)

Introduction Respiration -

Exchange of oxygen and carbon dioxide External respiration vs. Internal respiration Involuntarily or voluntarily controlled May limit metabolism

(Lindt et al., 2014; Prosser 1991)

Introduction Anaerobic vs. Aerobic Metabolism - Requirement of Oxygen - Both produce ATP, but differ in amounts - Oxidation of molecules

(Randall, 1997)

Introduction Calorimetry -

Measurement of heat in a system and surroundings Direct Calorimetry vs Indirect Calorimetry Measuring heat over a period of time Measuring other factors aside from heat ex. Oxygen consumption

(Sterling et al., 2013; Williams & Fruhbeck, 2009)

Materials and Methods

(400 ppm CO2, 20.9% O2)

Materials and Methods

𝜇𝐿 𝑔𝑎𝑠 1 𝑚𝐿 × 4.53 𝐿 𝑎𝑖𝑟 × 𝐿 𝑎𝑖𝑟 1000 𝜇𝐿 where ppm gas can either be the pressures of carbon dioxide or oxygen uL gas can either be the volume of carbon dioxide or oxygen

𝑉 = 𝑝𝑝𝑚𝑔𝑎𝑠 𝑓𝑖𝑛𝑎𝑙 − 𝑝𝑝𝑚𝑔𝑎𝑠 𝑖𝑛𝑖𝑡𝑖𝑎𝑙

Sample calculations: 𝑉 = 3822 𝑝𝑝𝑚 − 1642 𝑝𝑝𝑚

𝜇𝐿 1 𝑚𝐿 × 4.53 𝐿 × = 9.8754 𝑚𝐿 𝐿 1000 𝜇𝐿

Materials and Methods

755 𝑚𝑚𝐻𝑔 273 𝑉𝑐𝑜𝑟𝑟𝑒𝑐𝑡𝑒𝑑 = 𝑉𝑜𝑏𝑠𝑒𝑟𝑣𝑒𝑑 760 𝑚𝑚𝐻𝑔 273 + 𝑇 where T is the average temperature observed in °C

Sample calculations: 𝑉𝑐𝑜𝑟𝑟𝑒𝑐𝑡𝑒𝑑 = 9.8754 𝑚𝐿

755 𝑚𝑚𝐻𝑔 760 𝑚𝑚𝐻𝑔

273 = 8.950062 𝑚𝐿 273 + 26.24346

Materials and Methods

𝑉𝑂2𝑐𝑜𝑟𝑟𝑒𝑐𝑡𝑒𝑑 4.7𝑔𝑐𝑎𝑙 60 𝑚𝑖𝑛𝑠 10 𝑚𝑖𝑛𝑠 𝑚𝐿 𝑂2 1 ℎ𝑟 𝑀𝑅 = 𝑏𝑜𝑑𝑦 𝑤𝑒𝑖𝑔ℎ𝑡

Sample calculations: 10.3747 𝑚𝐿 𝑂2 4.7𝑔𝑐𝑎𝑙 10 𝑚𝑖𝑛𝑠 𝑚𝐿 𝑂2 𝑀𝑅 = 25.14 𝑔

60 𝑚𝑖𝑛𝑠 1 ℎ𝑟

= 11.6375 𝑔𝑐𝑎𝑙/ℎ𝑟/𝑔

Materials and Methods

𝑚𝐿 𝐶𝑂2 𝑅𝑅 = 𝑏𝑜𝑑𝑦 𝑤𝑒𝑖𝑔ℎ𝑡 × ℎ𝑟

Sample calculations: 𝑅𝑅 =

10.3747 𝑚𝐿 𝐶𝑂2 25.14 𝑔 × 0.17 ℎ𝑟

Results Concentration (ppm)

250000 200000

y = -3.311x + 206375 R² = 0.5266

150000

CO2 O2

100000 y = 3.4678x + 1657.4 R² = 0.9971

50000 0

0

200

400 Time (s)

600

800

Figure 1. Graph of changes in O2 and CO2 levels in the span of 600s.

Results Mouse

1

2

3

4

Weight (g)

28.88

33.66

25.14

29.5

Oxygen Consumption (mL/min)

32.4386

35.5499

10.3747

8.66581

CO2 Production (mL/min)

9.57926

10.4917

8.950062

10.00239

Metabolic Rate as Respiration Rate (mL/hr/g)

6.73932

6.33689

2.47606

1.76254

Metabolic Rate as Heat Production gcal/hr/g

31.6748

29.7834

11.6375

8.2839

Table 1. Weights, oxygen consumption, carbon dioxide production, and metabolic rates as respiration rate and heat production of four different mice.

Mass-specific Metabolic Rate as Respiration Rate (mL CO2/hr/g)

Results 2.15 2.1

2.05 2 1.95

y = -0.0307x + 2.9065 R² = 0.9446

1.9 1.85 0

10 20 30 Body Weight (g)

40

Figure 2. Graph of respiration rate vs. body weight .

Mass-specific Metabolic Rate as Heat Production (gcal/hr/g)

Results 35

30 25

y = 1.9217x - 35.952 R² = 0.3075

20 15

10 5 0 0

10 20 30 Body Weight (g)

40

Figure 3. Graph of heat production vs. body weight.

Discussion Levels of Metabolism Measuring Metabolism via Respiration Factors Affecting Metabolism

Levels of Metabolism 1. Standard metabolism 2. Routine metabolism 3. Active metabolism

Levels of Metabolism 1. Standard metabolism - resting, or fasting metabolism at a given temperature - absence of spontaneous activity, food digestion, and physical, thermal, or psychological stress. - ectotherms: SMR dependent on temperature - endotherms: BMR does not vary with temperature, but it must be measured in the thermoneutral zone (Randall et al. 2001; Willmer et al., 2005)

Levels of Metabolism 2.

3.

Routine metabolism - minimal normal unrestrained activity Active metabolism - distinguished by enforced maximal activity - maximum metabolism at a constant temperature

(Norin & Malte, 2011; Willmer et al., 2005; Harris et al., 2000)

Measuring Metabolism via Respiration Correction of oxygen volume using STP factors - to allow standardization and more accurate comparison of the results - the removal of pressure as a factor in calculating the metabolic rate - only to account for variations in gas levels and temperature (Campbell & Farrell, 2010)

Measuring Metabolism via Respiration Indirect calorimetry or respirometry 1. open - continuous flow of air - flow rate 1. closed - amounts of oxygen and carbon dioxide must theoretically decrease and increase, respectively, over time due to metabolic demands of respiration (Lighton, 2008)

Measuring Metabolism via Respiration Other methods of respirometry: 1. Douglas bag - respired air is collected for analysis of the changes in oxygen and carbon dioxide concentrations over a short period of time 1. Differential Scanning Calorimeters - ensuring the heat-flow rate to a sample organism against time or temperature and comparing it to the pre-programmed temperature of the sample in given atmospheric temperatures 1. OROBOROS Oxygraph-2k - allows for the instantaneous, and real time recording of oxygen consumption using new software (Williams & Fruhbeck, 2009; Höhne et al., 1996; Hütter et al., 2006)

Factors Affecting Metabolic Rate 1. 2. 3. 4. 5. 6. 7. 8.

Weight Sex Temperature Diet Light Hormone Levels Age Activity

(Randall, 1997; Frisancho, 2000)

Factors Affecting Metabolic Rate Effect of body weight on metabolic rate MR = a(body mass)^b where a is the intercept of the log regression line and b is the rate at which the metabolic rate changes by body mass As the weight of an organism increases, its metabolic rate decreases. Applicable only to different species as other factors play a bigger role (Chapman & Reiss, 1999; Randall, 1997)

Factors Affecting Metabolic Rate Metabolism and Surface Area - “Surface Area Law”

- Claim has been disregarded by most physiologists - implies that animals exhibiting relatively the same body temperatures would exhibit the same metabolic rates.

(Blaxter, 1989; Mitchell, 1962)

Factors Affecting Metabolic Rate Temperature - Different Effect for Ectotherms and Endotherms - Increase of oxygen consumption upon increase in temperature

(Randall et al., 2001; Schmidt-Nielsen, 1997)

Factors Affecting Metabolic Rate Activity - Increase of metabolism, oxygen consumption due to increase of locomotion velocity. - Activities that involve dormancy, such as hibernation lowers metabolic rate.

(Randall et al., 2001)

Factors Affecting Metabolic Rate Sex - Generally Females have 5-10% decrease in metabolic rate compared to Males - In humans females have a lower muscle mass relative to body size. Smaller metabolic rate per unit area - Effect of sex on metabolism is inconclusive, since effects become less pronounced as the individual ages. (Tarnopolsky, 1999; McArdie, 2000; Mahan et al. 2012)

Factors Affecting Metabolic Rate Light - Presence of light can affect the cycle of the animal’s activity. - Ex. A nocturnal animal may have a higher metabolic rate in the dark, as that is the state at which it is to be most active. - Yeast metabolism can be affected by certain wavelengths of light. (Summers, 2013; Robertson et al., 2013)

References Blaxter, K. 1989. Energy metabolism in animals and man. New York: Cambridge University Press. Brozek, J, 1965. Human Body Composition: Approaches and Applications. London: Pergamon Press. Chapman, J.L. and Reiss, M.J. 1999. Ecology Principles and Applications. Cambridge: Cambridge University Press. Data Studio [software]. Roseville, California: PASCO Scientific. Frisancho, A.R. 1993. Human Adaptation and Accommodation. United States of America: University of Michigan Press. Harris, R.P., Wiebe, P.H., Lenz, J., Skjoldal, H.R. and Huntley, M. 2000. Zooplankton Methodology Manual. London: Academic Press. Höhne, G., Hemminger, W., Flammersheim, H.J. Differential Scanning Calorimetry. Berlin: Springer-Verlag. Hütter, E., Unterluggauer, H., Garedew, A., Jasen-Dürr, P., and Gnaiger, E. 2006. High Resolution Respirometry-modern day tool in aging research. Experimental Gerentology. 46: 103-109 Lighton, J. (2008). Measuring Metabolic Rates: A Manual for Scientists. Oxford University Press: New York. 75-85. Lindh, W. Q., Pooler, M.S., Tamparo, C.D., Dahl, B.M., and Morris, J.A. 2014. Comprehensive Medical Assisting: Administrative and Clinical Competencies, 5th edition. New York: Cengage Learning. Mahan, L.K., Escott-Stump, S., Raymond, J. 2012. Krause’s Food and Nutrition Care Process. United States of America: Elsevier. McArdie, W.D. 2000. Essentials of Exercise Physiology. United States of America: Lippincott Williams & Wilkins. Mitchell, H. 1962. Comparative Nutrition of Man and Domestic Animals. United Kingdom: Academic Press, Inc. Norin, T., and Malte, H. 2011. Reliability of standard metabolic rate, active metabolic rate, and aerobic scope in young brown trout during period of moderate food availability. Journal of Experimental Biology. 214 1668-1675 Prosser, C. L. 1991. Comparative Animal Physiology: Environmental and Metabolic Animal Physiology, 4th edition. Massachusetts: John Wiley & Sons, Inc. Randall, D., Buggren, W., and French, K. (1997). Eckert Animal Physiology: Mechanisms and Adaptations. 4th ed. New York: W. H. Freedman and Company.

References Reece, J. B., Urry, L. A., Cain, M. L., Wasserman, S. A., Minorsky, P. V., & Jackson, R. B. 2010. Campbell Biology. San Francisco: Pearson Benjamin Cummings. Robertson, J.B., Davis, C.R., and Johnson, C.H. 2013. Visible light alters metabolic rhythms by inhibiting respiration. PNAS. 110(52): 21130-21135 Schmidt-Nielsen, K. 1997. Animal Physiology: Adaptation and Environment, 5th edition. New York: Cambridge University Press Sterling, E.J., Bynum, N., and Blaire, M.E. 2013. Primate Ecology and Conservation. United Kingdom: Oxford University Press. Summers, M. 2011. Marine Respiration: The Effects of Temperature, Light, and Body Size on Pacific Zooplankton and Reef Goby Collected 2-10ºN Latitude. Stanford University. Tarnopolosky, M. 1999. Gender Differences in Metabolism. United States of America: CRC Press. Williams, G. and Fruhbeck, G. 2009. Obesity: Science to Practice. United Kingdom: John & Wiley Sons, Ltd. Willmer, P., Stone, G. and Johnston, I. 2005. Environmental Physiology of Animals. Massachusetts: Blackwell Publishing.