Digestibility measurement system

Carbohydrate and fat primarily serve as the energy source (Ravindran & Bryden, 1999) supplying energy needed by the animal. Protein provides energy through oxidation and gluconeogenesis, but primarily supplies AA (Ravindran & Bryden, 1999). These two main functions of food in animals give rise to two digestibility measurement systems: the energy evaluation systems, and AA digestibility systems.

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2.8.2.1 Energy systems

The three different energy systems (see Figure 2.5) used in determining the energy requirements of an animal include digestible energy (DE), metabolisable energy (ME), and net energy (NE). These all use gross energy as the basis of their calculation.

Gross energy (GE) is the energy content of the feed, normally determined by measuring the heat of combustion in a bomb calorimeter when a feed sample is completely oxidised. It forms the basis from which digestible energy is calculated. DE is the amount of energy that disappears when an animal eats the feed. That is, the GE of feed minus the GE of faeces. It assumes that the energy that disappears when ingested is digested, absorbed and available for use by the animal (McDonald et al., 2011). It is determined by recording total feed intake and total faecal output, measuring their GE content, and subtracting faecal energy from intake energy, that is: DE = IE – FE (Noblet & Henry, 1993). An alternative method is the inclusion of indigestible markers (Boisen & Fernandez, 1997). DE is the simplest method of measuring digestibility, but it fails to take into consideration energy losses through urine and combustible gasses during feed metabolism. ME is calculated by subtracting all the energy lost as waste in the excretion of urine and production of combustible gasses from DE (Pond et al., 1995). The energy lost through urine is measured by collecting urine and analysing it for energy content. Energy lost through combustible gasses is measured by keeping the animal in a respiration chamber where the amount of gasses produced is collected. The values obtained are used to determine ME using the following formula (Boisen & Fernandez, 1997; McDonald et al., 2011; Noblet & Henry, 1993).

ME = DE – (urinary energy + energy in combustible gases)

This method of measuring digestibility gives a better estimate of energy available to the animal as it corrects the DE for efficiency of protein use, however; it involves more procedures than DE.

NE describes the energy that is actually used by the animal and is obtained from subtracting heat increment produced during digestion of feed, metabolism of nutrients, and excretion of waste from ME. It is the amount of energy actually used for maintenance and production (growth, lactation and gestation) and is calculated using the following equation (McDonald et al., 2011; Pond et al., 1995):

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Figure 2.5 Three Energy Evaluation Systems.

Source: Leeson & Summer, 2001.

The amount of energy produced during heat loss can be obtained from adiabatic or gradient layer calorimetry. In adiabatic calorimetric studies, the animal is fed and placed in a shield chamber filled with a weighed volume of water and a bomb calorimeter. The chamber is shielded with an adiabatic shield that serves as a heat insulator. The heat of combustion produced from the animal’s body during digestion and metabolism of feed causes an increase in the level of water. This increase in water level over time is measured by the adiabatic bomb calorimeter, which indicates the amount of heat produced. In gradient layer calorimetry, heat produced passes through the chamber wall and is measured by the flow of heat from it. An alternative method includes calculation of oxygen consumption and excretion of NH4, CO2 and N using the following equation proposed by several

researchers (Boisen & Fernandez, 1997; McDonald et al., 2011; Noblet & Henry, 1993). Heat Production = 3.866 x O2 + 1.2 x CO2 – 0.518 x NH4 – 1.431 x N

It is the most accurate and unbiased way of characterising energy content of feed. However, it is much more difficult to determine, as it is more complex than DE and ME.

Metabolisable Energy (ME)

Ingested Energy (IE) or Gross Energy (GE) Faecal

Digestible Energy (DE) Urinary Energy

Methane Energy/ Combustible gas

Heat Increment from microbialfermentation and obligatory thermogenesis, i.e. excess heat relative to glucose during ATP synthesis

Net Energy

Production Energy:

Growth, Milk Production, Egg Production, etc

Maintenance:

Basal metabolism, physical activity, bodytemperature regulation, etc

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2.8.2.2. Methods of measuring amino acid digestibility

Energy systems indicate the distribution and absorption of energy from feed. Protein and AA are other factors to be considered in formulating feed for the animals. The accurate measurement of these values in feed and their digestibility in the animal is necessary for maintaining optimum growth and performance. These values are determined using three methods of measurement including apparent faecal digestibility, apparent ileal digestibility, and true ileal digestibility.

Apparent faecal digestibility (AFD) is the procedure used in evaluating digestibility of dietary protein based on the difference between the amount ingested and the amount appearing in faeces. Apparent protein digestibility can be determined as the difference between the amount of N ingested and excreted, expressed as a proportion of N ingested. AFD can also be used to determine digestibility of a specific AA when the concentration of that AA in feed and faeces are measured. It is calculated as: AA digestibility = AA in feed – AA in faeces / AA in feed. AA in faeces is the total AA from undigested diet residues and endogenous secretion; hence, it underestimates the value of digestible AA (McDonald et al., 2011). The amount of AA in feed is measured before feeding, and all faeces are collected and analysed after egestion for AA content (Low, 1982).

Apparent ileal digestibility (AID) is determined by measuring the quantity of AA remaining at the end of the small intestine or ileum. This provides a much more accurate indication of protein digestibility and availability by the animal because the problems associated with microbial protein degradation in the large intestine and caecum does not arise. McDonald et al., (2011) suggest that microbial activity in the small intestine may possibly affect the accuracy of apparent ileal digestibility values, however, it seems reasonable to assume that any affect would be minimal compared with that observed in the large intestine (McDonald et al., 2011). AID is calculated using the following equation:

AID% = AA intake – ileal AA outflow X 100 AA intake

There are several methods available to collect ileal digesta for measuring apparent ileal digestibility. Sampling from the terminal ileum after euthanasia is common in both rats and pigs, whereas other methods, such as cannulation and bypass of the large intestine by ileorectal anastomy or ileostomy, are more commonly performed on pigs. Each of these methods have their own advantages and disadvantages involved (Moughan, et al., 2000).

True ileal digestibility (TID) measures the proportion of dietary AA that disappears from the digestive tract before the distal ileum and does not include endogenous AA. It is a fundamental

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property of the food that is not affected by the dietary conditions under which the food is fed to the animal (e.g., the protein content of the test diet). The apparent digestibility measure, however, is affected by the assay conditions and is, therefore, variable and subject to error. True digestibility is a superior measure for determining the AA that are absorbed from the gut, and therefore, gives a better representation of protein quality than apparent digestibility (Stein et al., 2007). It is calculated using the following equation:

TID % = AA intake – (ileal AA outflow – total IAA end) X 100 AA intake

It is determined by subtracting the amount of AA in ileal digesta from the amount of AA that were ingested, to obtain an “apparent” digestibility coefficient. However, ileal digesta also contains a significant proportion of non-dietary AA, from sources such as mucus, cells, digestive enzymes, and bile known as endogenous amino acids loss (EAAL) (Moughan et al., 2000). These EAAL can be quantified using several methods. One method is by feeding a protein-free diet to the animal with the protein remaining at the end of the small intestine assumed to be the basal EAAL. The provision of a diet devoid of protein, however, leads to negative body nitrogen balance in the recipient animal (Moughan et al., 2000). Another method is to feed the test animal an enzyme hydrolysed protein (usually enzyme hydrolysed casein) diet with ultrafiltration of the ileal digesta collected to remove any unabsorbed dietary AA. It is assumed any protein at the terminal ileum is from endogenous sources. When the basal EAAL is determined using techniques such as the protein-free method or the enzyme hydrolysed protein method, apparent AA digestibility is corrected in the calculation of digestibility, and the “true” digestibility coefficient is obtained (Moughan et al., 2000).