EXERCISE 5

Measuring Carbon Dioxide Production in Aquatic Animals

Valerie Sol T. Bengzon

University of the Philippines Visayas Tacloban College

Division of Natural Science and Mathematics

INTRODUCTION

Gas exchange is the exchange of oxygen and carbon dioxide between the

animal’s body fluids and the environment it thrives in. As an animal breathes, it takes

oxygen from the environment which is used for ATP production, also called anaerobic

respiration. Carbon dioxide production and released back into the environment is also

associated with this. Virtually all animals depend on such aerobic metabolism to satisfy

their resting energy requirements, although it is possible to produce energy, in the

form of ATP, by anaerobic metabolism (Kay 1998). The metabolism of carbohydrates

and fats, two of the major nutritional components of animals, results in the production

of carbon dioxide and water as waste products. Carbon dioxide is removed via

exhalation from the lungs or ventilatory structures in various animals. The exchange

of gases between animals and their environments occurs by the process of simple

diffusion. For many animals, small aquatic animals in particular, the exchange of gases

across the general body surface is sufficient to meet the demands of the animal.

Aerobic respiration, also known as aerobic metabolism in animals, occurs when

oxygen is taken in by the body and sent into its cells which is later then used for energy

production. Animals undergo the process differently using different respiratory

structures. Ectotherms such as fish are animals that depend on the environment for

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body temperature. All aquatic ectotherms adapt to their environment in different ways.

They can regulate their body temperature through adapting their eating habits or

physical endurance. In terms of cost to the animal, though, this is not necessarily so.

If an animal is ectothermic, in metabolic terms, it costs the animal nothing in order to

maintain some sort of appropriate body temperature for at least part of the day.

In this exercise, the carbon dioxide production of a fish was investigated.

Specifically, the study aimed to estimate and compare the rate of carbon dioxide in

small aquatic animals such as fish by titrimetric method; and to relate carbon dioxide

production with metabolic rate in aquatic animals.

MATERIALS AND METHODS

Three set ups of two 250 ml beakers, with varying temperatures at 23C, 28C,

33C, were labelled with A and B were filled with distilled water treated with 5 drops of

phenolphthalein. In each set-up, a medium sized Shobon, a hybrid of koi and goldfish,

was added in beaker A while beaker B served as the control wherein it did not contain

any animal. After 30 minutes upon starting the test, the fish in beaker A of each set-

up was removed. The carbon dioxide content of the water in each beakers were

determined. Using a pipette, 0.04% NaOH was slowly added into the beakers. Enough

NaOH was added until the water turns pink for one minute after swirling. The volume

of NaOH used in the titration was recorded and multiplied to 10 since each ml of 0.04%

NaOH solution combines with 10 μM of carbon dioxide. The weight of the fish used for

each set-up were weighed using an analytical balance. The respiration rate for each

temperature was computed using the equation:

𝜇𝑀 𝐶𝑂2 𝑜𝑓 𝐴 − 𝜇𝑀 𝐶𝑂2 𝑜𝑓 𝐵
𝑅𝑒𝑠𝑝𝑖𝑟𝑎𝑡𝑖𝑜𝑛 𝑟𝑎𝑡𝑒 (𝜇𝑀 𝐶𝑂2)/𝑔/ℎ𝑟 =
𝑤𝑒𝑖𝑔ℎ𝑡 (𝑔)𝑥 𝑡𝑖𝑚𝑒 (ℎ𝑟)

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 RESULTS AND DISCUSSION

Ectotherms such as fish are animals dependent upon external sources for heat

gain. Little of their body heat is obtained as a consequence of their overall metabolism.

In the exercise, the metabolic rate was determined based on the respiratory gas

exchange, which is the amount of oxygen consumed and carbon dioxide produced in

metabolism. Carbon dioxide production was specifically used to measure the

metabolic rate. Water has a carbon dioxide solubility 20-30 times greater than that of

the air (Kay 1998). It can dissolve in water 200 times more easily than oxygen. Thus,

aquatic animals such as the Shobon fish have very little problem in excreting the

carbon dioxide produced during aerobic metabolism.

Table 1. Effect of Temperature on the Respiratory Rate of the Shobon Fish

Temperature Fish Weight 0.4 NaOH volume (mL) Respiration

20
Respiration rate (μM CO2)/g/hr

15
15.63
10

0 2.91

-5
-5.25
-10
23 28 33
Temperature C

Figure 1. Trend of the respiratory rate against temperature of the Shobon fish

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 Metabolic rate is the measurement of the sum total of all metabolism occurring

in an animal at a given time. This can be measured in different ways such as oxygen

consumption, heat production and carbon dioxide production. Oxygen is used up in

cellular respiration, and carbon dioxide is produced as a by-product. For an ectotherm

such as fish, the standard metabolic rate varies with temperature. In the exercise, the

metabolic rate or respiratory rate of the Shobon fish was determined using the carbon

dioxide production at varying temperatures. The fish exhibited different rates of

respiration wherein the highest was at 33C and the least at 23C. Water temperature

is one of the most important environmental variables influencing the metabolic rates

of ectotherms (Breth 1971). In its natural environment, fishes may experience intense

water temperature variations on a daily basis and can fluctuate by up to 10-15C daily.

Warm water temperature increases the metabolism of the fish and therefore

respiration rates are high. In all animals, a higher rate of physical activity oxidizes more

calories. In fish, whose metabolic rate is affected by the oxygen content of the water,

the oxygen effect interacts with the activity effect. As the rate of activity increases, the

fish needs more oxygen in the water to sustain aerobic metabolism.

In aquatic ectotherms such as the Shobon fish, metabolism is affected by

temperature. Water temperature influences the body temperature of aquatic

ectotherms. Heat lose by evaporation is not possible and heat exchange by radiation

is reduced in water since water is an effective absorber of infrared radiation. Animals

get the energy to sustain life by oxidizing reduced carbon compounds in food,

therefore metabolic rate can be measured experimentally by determining an animal's

rate of oxygen consumption and carbon dioxide production. The standard metabolic

rate is the metabolic benchmark for fish. In poikilotherms, this is translated into

biochemical events and in turn into metabolism. The metabolic rates of fish roughly

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double with every 10C increase in temperature, except at the extreme ends of their

temperature tolerance. At 23C, which is relatively lower than the room temperature,

had the least metabolic rate. As the temperature decreases, the respiratory rate

decreases so as the metabolic rate. As the temperature increases in 5C increments

to 28C and 33C, the metabolic rate increases also. The higher metabolic rates in

higher temperatures can be attributed to the oxygen availability in water production of

carbon dioxide. For every milliliter of oxygen consumed, 0.9 milliliters of carbon dioxide

is produced. In low temperature, the fish consumed less oxygen therefore less carbon

dioxide is produced than fish acclimated in higher temperatures.

Metabolic rate does not only vary as temperature fluctuates. It can vary

depending on physiological states. There is a correlation between the metabolic rate

and body size ratio in animals. As a general rule, the greater the mass of an organism,

the higher is the organism’s metabolic rate. Animals with high metabolic rates require

more efficient delivery of oxygen to cells. However, metabolic rate is higher per unit of

body mass in small animals compared to larger ones. The surface rule states that the

metabolic rate per unit weight decreases with increasing size, but is constant per unit

surface (Bertalanffy 1951). This is because the higher metabolic rate of small animals

needs a greater delivery of oxygen to tissues around the body. In the exercise, data

obtained could not support such correlation since no set-ups in varying fish weight

were subjected to just one temperature. Rather each set-up with different fish weights

were subjected to different temperature, thus, fish weight alone could not be the sole

basis for the varying metabolic rates. In fish, a major area of metabolic heat loss is via

the gills. This also means that some regions are kept at a different temperature from

other regions which is advantageous to the fish because the elevated temperature

means an increased rate of metabolic reactions in those parts. An endotherm is

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dependent upon the internal heat production and derives its heat gain primarily from

cellular metabolism. Increased muscular activity as well as hormone activities,

increase the metabolic heat production. The possible errors in the exercise would be

the lack of acclimatization of the fish to the varying temperatures. The fishes used

were directly subjected to the temperature without allowing them to acclimatize. The

fish’s health was not also ensured to be at its best, thus its metabolic rate could be

affected due to the inability or weaker response to the stimulus. Another factor could

also be attributed to the determination of carbon dioxide production. There could be

possible errors incurred during the titration process.

In the aerobic catabolism of carbohydrates, lipids and proteins, corresponding

heat production per ml of carbon dioxide produced in grams calories are 5.05, 6.67

and 5.57 respectively. The metabolic rate of the animals in terms of heat production

depending on the type of animal. Ectotherms are animals dependent upon external

sources for heat gain, however, in aquatic ectotherms large bodies of water provide a

relatively stable environment in terms of temperature. Water as an effective heat sink

means that metabolic heat generated by the organism may be dissipated into the

water and rapid heat loss occurs.

Three metabolic rates have been distinguished in respect to the relation

between metabolic rate and body size. The first type, surface-dependent, metabolic

rate is proportional to a surface or two-thirds power of weight. The rate of oxygen

consumption decreases per unit weight, but is constant per unit surface, expressed as

the % power of weight. Representatives of this type are fish, crustaceans, mussels

and Ascaris (Bertalanffy and Muller 1943; Weinland 1919; Kruger 1940). The second

type, weight-dependent, metabolic rate is directly proportional to weight. A

representative of this type is an insect larvae (Bertalanffy and Muller 1943). The third

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type, intermediate between weight proportionality to proportionality to surface.

Metabolic rates decrease with respect to weight, but increase with respect to surface.

A representative of this type is pond snails (Bertalanffy and Muller 1943). Also, the

smaller animals have a greater surface area to volume ratio, so more heat is lost.

CONCLUSION

Aquatic ectotherms, such as the Shobon fish, undergo various energy

consuming physiological processes liberating heat to reflect the animal’s metabolic

rate. During gas exchange, oxygen consumption and carbon dioxide production have

calorific or heat equivalents. Metabolic rate can be measured by oxygen consumption,

carbon dioxide production and heat production. Using the titrimetric method to

determine the carbon dioxide production rate, the Shobon fish illustrated that an

increase in carbon dioxide production is an increase in its metabolic rate. Temperature

is also a factor that influences the metabolic rate of fish wherein an increase in

temperature illustrated an increase in the fish’s metabolic rate.

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Bertalanffy, L.V. 1951. Metabolic Types and Growth Types. The American Naturalist.

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