# Direct and Indirect Calorimetry

Direct and indirect calorimetry:

1. Describe different types of calorimetry? (2018)
2. Direct Vs Indirect calorimetry

Direct and indirect calorimetry: Intensity of energy metabolism can be estimated either by calculating heat production from the exchange of respiratory gases (indirect calorimetry) or by measuring the heat which is lost directly from the body by radiation, conduction, convection and evaporation (direct calorimetry).

Direct Calorimetry:

Principle: Direct calorimetry measures heat production directly by detecting and quantifying the heat generated by an animal.

Measurement: Direct calorimetry typically involves the use of a calorimeter, a specialized chamber or device that isolates the animal and measures the temperature change of the surrounding air or water. As the animal undergoes metabolic processes, such as digestion, respiration, and physical activity, heat is generated and detected as a change in the temperature of the calorimeter.

• Adiabatic calorimetry involves placing an animal in a chamber designed to minimize heat loss through its walls.
• This is achieved by having an outer box or wall that is electrically heated to the same temperature as the inner wall, preventing heat loss from the inner wall to the outer wall.
• Water circulating in a coil within the chamber absorbs the heat collected by the inner wall, and changes in the volume and temperature of the water can be used to calculate sensible heat loss from the animal’s body.

• Gradient calorimetry allows for some heat loss through the walls of the animal chamber.
• The outer surface of the wall of the calorimeter is maintained at a constant temperature with a water jacket, and temperature gradients are measured with thermocouples lining the inner and outer surfaces of the wall.

1. Precision and Accuracy: Direct calorimetry provides precise and accurate measurements of heat production because it directly measures the heat generated by the subject.
2. Useful for Respiratory Studies: Direct calorimetry can be combined with measurements of oxygen consumption (VO2) and carbon dioxide production (VCO2) to assess respiratory parameters, such as the respiratory quotient (RQ).
3. Quantitative Data: Direct calorimetry provides quantitative data in units of heat energy (e.g., calories or joules).
4. Measurement of Insensible Heat: Direct calorimetry estimates insensible heat loss, including the latent heat of water vaporized from the skin and respiratory passages.
5. Versatile Applications: Can be used for various organisms, from small animals to humans.
6. Thermoregulation Insights: Provides data on evaporative heat loss, useful for understanding thermoregulation.

1. High Cost: The construction and maintenance of direct calorimeters, especially adiabatic calorimeters and gradient calorimeters, can be expensive.
2. Confinement of Subjects: Subjects must remain in a confined environment within the calorimeter for extended periods to obtain accurate measurements.
3.  Limited Real-World Validity:  Artificial conditions may not reflect natural metabolic rates.
4. Time-Consuming: Requires prolonged observation periods for accurate measurements.

Indirect Calorimetry:

Principle: Indirect calorimetry estimates heat production indirectly by measuring the consumption of oxygen (O2) and the production of carbon dioxide (CO2) during metabolic processes. It is based on the fact that the combustion of carbohydrates, fats, and proteins in the body requires O2 and produces CO2, and the amount of O2 consumed and CO2 produced is related to the energy expended.

Measurement: Indirect calorimetry is typically performed using specialized equipment, such as metabolic carts or gas analyzers. Because the animal body ultimately derives all of its energy from oxidation, the magnitude of energy metabolism can be estimated from the exchange of respiratory gases. Varieties of techniques are available for measuring the respiratory exchange; all ultimately seek to measure oxygen consumption and CO2 production per unit of time. Thus, Heat loss is measured indirectly by gaseous exchange: CO2 output/O2 intake called Respiratory quotient (RQ).

The RQ serves as an indicator of the type of substrate being oxidized, such as lipid or carbohydrate, during the metabolic processes. Carbs have an RQ of 1 while fat have an RQ of 0.7 as they require more oxygen.

Types of Indirect Calorimetry:

Closed Circuit System:

• In a closed circuit system an animal, rebreathes the same air within a closed chamber.
• CO2 is removed with a suitable absorbent, which may be weighed before and after use to determine its rate of production.  The change in the volume of the respiratory gas mixture due to oxygen consumption is used to estimate oxygen use.
• Oxygen consumed by the animal is replaced by a metered supply of pure oxygen.
• Corrections are made for any differences in the amounts of O2 and CO2 present in the circuit air at the beginning and end of the experiment. Methane, if produced, can accumulate in the circuit air, and its amount is determined at the end of the experiment.

Open Circuit System:

• In an open circuit system, an animal is provided with a specific air composition, and the outgoing air from a chamber or expelled air from a mask is either collected or measured and sampled.
• Collected gas is then analyzed to determine its oxygen and carbon dioxide content using various techniques, including chemical, volumetric, or nanometric methods.

Respiratory Quotient (RQ): CO2 output/O2 intake

• The RQ is a critical parameter in indirect calorimetry, and it quantifies the ratio of carbon dioxide produced (VCO2) to oxygen consumed (VO2) during metabolism.
• The RQ values differ for various nutrients: For carbohydrates, the RQ is 1 (6 CO2 produced for every 6 O2 consumed). For fats, the RQ is approximately 0.70 (51L CO2 produced for every 72.5L O2 consumed). For proteins, the RQ is approximately 0.8 (0.77L of CO2 produced for every 0.96L of O2 consumed per gram of protein oxidized).
• The RQ provides information about the type of substrate being oxidized for energy, as well as the proportion of oxygen used for each nutrient.

C6H12O6 + 6 O2 → 6 CO2 + 6 H2O + 2820 KJ

1. Non-Invasive: Indirect calorimetry is a non-invasive method that does not require any surgical procedures or the insertion of probes into the body.
2. Objective Assessment: It offers an objective and quantitative assessment of metabolic processes.
3. Substrate Utilization: Indirect calorimetry allows for the determination of which substrates (carbohydrates, fats, or proteins) are being utilized for energy production.

1. Equipment Costs: High-quality indirect calorimetry equipment can be expensive to purchase and maintain.
2. Complexity: The operation and calibration of indirect calorimetry equipment can be complex, requiring specialized training and expertise.
3. Environmental Factors: Environmental conditions, such as temperature and humidity, can influence gas exchange measurements and require careful control during experiments.
4. Measurement of Protein Oxidation: Indirect calorimetry is less accurate in estimating protein oxidation compared to carbohydrate and fat oxidation.
1. How energy retention in the animal body is measured by Carbon- nitrogen balance study ? How does it differ from that of comparative slaughter methods? (2017)
2. Differentiate between metabolic faecal nitrogen and endogenous urinary nitrogen ? ( 2018)

### Practice Questions:

1. Explain the principles of direct calorimetry and indirect calorimetry. How do these methods differ in measuring the energy expenditure of animals?
Marks: 10
2. Describe the equations used in indirect calorimetry to estimate heat production. Discuss how oxygen consumption, carbon dioxide production, methane, and nitrogen excretion are used in these calculations.
Marks: 10