ANIMAL NUTRITION

UNIT-1
The animal and its food
1.1 Water
1.2 Dry matter and its components
1.3 Analysis and characterisation of foods
Food is material that, after ingestion by animals, is capable of being digested, absorbed and
utilised. In a more general sense we use the term ‘food’ to describe edible material. Grass and
hay, for example, are described as foods, but not all their components are digestible. Where
the term ‘food’ is used in the general sense, as in this book, those components capable of
being utilised by animals are described as nutrients.
The animals associated with humans cover the spectrum from herbivores, the plant eaters
(ruminants, horses and small animals such as rabbits and guinea pigs); through omnivores,
which eat all types of food (pigs and poultry); to carnivores, which eat chiefly meat (dogs and
cats). Under the control of humans these major classes of animal still pertain, but the range of
foods that animals are now offered is far greater than they might normally consume in the
wild (for example, ruminants are given plant by-products of various human food industries
and some dog foods contain appreciable amounts of cereals).
Nevertheless, plants and plant products from the major source of nutrients in animal nutrition.
The diet of farm animals in particular consists of plants and plant products, although some
foods of animal origin such as fishmeal and milk are used in limited amounts. Animals
depend upon plants for their existence and consequently a study of animal nutrition must
necessarily begin with the plant itself.
Plants are able to synthesise complex materials from simple substances such as carbon
dioxide from the air, and water and inorganic elements from the soil. By means of
photosynthesis, energy from sunlight is trapped and used in these synthetic processes. The
greater part of the energy, however, is stored as chemical energy within the plant itself and it
is this energy that is used by the animal for the maintenance of life and synthesis of its own
body tissues. Plants and animals contain similar types of chemical substances, and we can
group these into classes according to constitution, properties and function. The main
components of foods, plants and animals are:
1.1 WATER: The water content of the animal body varies with age. The newborn animal
contains 750–800 g/kg water but this falls to about 500 g/kg in the mature fat animal. It is
vital to the life of the organism that the water content of the body be maintained: an
animal will die more rapidly if deprived of water than if deprived of food.
Water functions in the body as a solvent in which nutrients are transported about the
body and in which waste products are excreted. Many of the chemical reactions
brought about by enzymes take place in solution and involve hydrolysis. Because of
the high specific heat of water, large changes in heat production can take place within
the animal with very little alteration in body temperature.
Water also has a high latent heat of evaporation, and its evaporation from the lungs
and skin gives it a further role in the regulation of body temperature.The animal
obtains its water from three sources: drinking water, water present in its food, and
metabolic water, this last being formed during metabolism by the oxidation of
hydrogen-containing organic nutrients.
The water content of foods is variable and can range from as little as 60 g/kg in
concentrates to over 900 g/kg in some root crops. Because of this great variation in
water content, the composition of foods is often expressed on a dry matter basis,
which allows a more valid comparison of nutrient content.
This is illustrated in Table 1.1, which lists a few examples of plant and animal
products. The water content of growing plants is related to the stage of growth, being
greater in younger plants than in older plants. In temperate climates the acquisition of
drinking water is not usually a problem and animals are provided with a continuous
supply. There is no evidence that under normal conditions an excess of drinking water
is harmful, and animals normally drink what they require.
1.2 DRY MATTER AND ITS COMPONENTS: The dry matter (DM) of foods is
conveniently divided into organic and inorganic material, although in living organisms
there is no such sharp distinction. Many organic compounds contain mineral elements as
structural components.
Proteins, for example, contain sulphur, and many lipids and carbohydrates contain
phosphorus. It can be seen from Table 1.1 that the main component of the DM of
pasture grass is carbohydrate, and this is true of all plants and many seeds. The
oilseeds, such as groundnuts, are exceptional in containing large amounts of protein
and lipid material.
In contrast, the carbohydrate content of the animal body is very low. One of the main
reasons for the difference between plants and animals is that, whereas the cell walls of
plants consist of carbohydrate material, mainly cellulose, the walls of animal cells are
composed almost entirely of lipid and protein.
Furthermore, plants store energy largely in the form of carbohydrates such as starch
and fructans, whereas an animal’s main energy store is in the form of lipid. The lipid
content of the animal body is variable and is related to age, the older animal
containing a much greater proportion than the young animal. The lipid content of
 living plants is relatively low, that of pasture grass, for example, being 40–50 g/kg
DM. In both plants and animals, proteins are the major nitrogen-containing
compounds.
In plants, in which most of the protein is present as enzymes, the concentration is high
in the young growing plant and falls as the plant matures. In animals, muscle, skin,
hair, feathers, wool and nails consist mainly of protein. Like proteins, nucleic acids
are also nitrogen-containing compounds and they play a basic role in the synthesis of
proteins in all living organisms. They also carry the genetic information of the living
cell.
The organic acids that occur in plants and animals include citric, malic, fumaric,
succinic and pyruvic acids. Although these are normally present in small quantities,
they nevertheless play an important role as intermediates in the general metabolism of
the cell. Other organic acids occur as fermentation products in the rumen, or in silage,
and these include acetic, propionic, butyric and lactic acids.
Vitamins are present in plants and animals in minute amounts, and many of them are
important as components of enzyme systems. An important difference between plants
and animals is that, whereas the former can synthesise all the vitamins they require for
metabolism, animals cannot, or have very limited powers of synthesis, and are
dependent upon an external supply.
The inorganic matter contains all those elements present in plants and animals other
than carbon, hydrogen, oxygen and nitrogen. Calcium and phosphorus are the major
inorganic components of animals, whereas potassium and silicon are the main
inorganic elements in plants.
1.3 Proximate analysis of foods: This system of analysis divides the food into six fractions:
moisture, ash, crude protein, ether extract, crude fibre and nitrogen-free extractives.
The moisture content is determined as the loss in weight that results from drying a
known weight of food to constant weight at 100 °C. This method is satisfactory for
most foods, but with a few, such as silage, significant losses of volatile material
(short-chain fatty acids and alcohols) may take place.
The ash content is determined by ignition of a known weight of the food at 550 °C
until all carbon has been removed. The residue is the ash and is taken to represent the
inorganic constituents of the food. The major component of ash is silica but ash may,
however, contain material of organic origin such as sulphur and phosphorus from
proteins, and some loss of volatile material in the form of sodium, chloride,
 potassium, phosphorus and sulphur will take place during ignition. The ash content is
thus not truly representative of the inorganic material in the food either qualitatively
or quantitatively. Animals do not have a requirement for ash as per, but require the
individual mineral elements that it contains.
Crude protein is the amount of protein of animal feed or specific food. Crude protein
depends on the nitrogen content of the food proteins. Crude protein measurements are
common in fields of animal husbandry and food science. It leads to economical
implications. Soybean remains the most important and preferred source of high
quality vegetable protein for animal feed manufacture. Soybean meal, which is the by-
product of oil extraction, has a high crude protein content of 44 to 50 percent and a
balanced amino acid composition, complementary to maize meal for feed formulation.
A high level of inclusion (30-40 percent) is used in high performance monogastric
diets.
The ether extract (EE) is a portion of dry matter extracted with ether. It is a
laboratory test to approximate the total fat (or crude fat) content of a feed and includes
some waxes, pigments and other lipids to a minor degree in addition to true fats.
The carbohydrate of the food is contained in two fractions, the crude fibre (CF) and
the nitrogen-free extractives (NFE).The former is determined by subjecting the
residual food from ether extraction to successive treatments with boiling acid and
alkali of defined concentration; the organic residue is the crude fibre.
Crude fibre estimates the indigestible fraction of feed or those fractions of the feed that are
fermented in the hindgut by microbes. Crude fibre is made up of plant cell structural
components, including cellulose, hemi-cellulose, lignin and pectin. For non-ruminant
animals, crude fibre is of little value energy-wise.
As determined by the customary procedure for the analysis of feeding stuffs nitrogen-free
extract is the largest component of the rations of animals, representing 40-70 per cent of the
total dry matter. It serves as a source of energy for body processes and for the deposition of
fat.
Digestive System of Ruminants

Ruminant animals are set apart from other mammals by their complex digestive systems.
The way they process food, absorb nutrients and gain energy is different from other
herbivores. The main distinction in a cow’s digestive system or a ruminant digestive system
is that the stomach has four separate compartments, each with a unique function, whereas
most other animals only have a single compartment with a unified functionality.
The four compartments allow ruminant animals to digest grass or vegetation without
completely chewing it first. Instead, they only partially chew the vegetation, then
microorganisms in the rumen section of the stomach break down the rest. Animals with
singular stomach compartments known as a monogastric digestive system do not have the
same capability.
Many different animals have this unique four compartment stomach type of digestive system,
including:
 Cattle
 Sheep
 Goats
 Water buffalo
 Deer
 Elk
 Giraffes
 Camels
These animals convert plant matter and vegetation into useable energy more efficiently than
other herbivores.
In cattle and dairy cows, the development, pH balance, functionality and bacteria levels of the
digestive system are crucial to maintaining overall health and high yield.
While some parts of the ruminant digestive system are similar to those of non-ruminant
systems, several essential components perform the necessary functions for digestion.
Components of the Ruminant Digestive System
 Mouth
 Esophagus
 Stomach
 Small Intestine
 Cecum
 Large Intestine

The Four Components of a Cattle’s Stomach

 Rumen
 Reticulum
 Omasum
 Abomasum
Components of the Ruminant Digestive System
While the ruminant digestive tract operates differently from the monogastric system, it is
composed of the same six basic components:
1. Mouth: The mouth is where the process begins. Cattle will graze by wrapping their
tongues around plants and tearing, pulling them into their mouth for mastication. They chew
first with the lower jaw incisors, working against a hard dental pad on the front part upper
palate, then second with the molars, grinding plant material down further. Chewing
stimulates saliva production and the saliva mixes with plant matter before the animal
swallows. Saliva contains enzymes capable of breaking down fats and starches and helps to
buffer the pH levels in the reticulum and rumen segments of the stomach. Mature cattle will
swallow from 50 to 80 quarts daily to aid in digestion, but the amount varies based on how
much time they spend chewing.
2. Esophagus: When the cattle swallow the plant material and saliva mix, it will travel down
the esophagus to the rumen. The esophagus performs the swallowing action through waves of
muscle contractions, moving the feed down. It has a bidirectional function, meaning it can
move feed from the mouth to the stomach or from the stomach to the mouth. Cattle need the
latter to regurgitate “cud,” or the under-chewed plant matter and grain, back up to the mouth
for further grinding. Once the cow is finished chewing the cud, it again swallows the matter
back down to the stomach.
3. Stomach: Generally, the stomach functions to further break down plant matter and grain.
More specifically, there are four sections of the stomach — rumen, reticulum, omasum and
abomasum — each with a particular job to do. These sections store chewed plant material and
grain, absorb nutrients and vitamins, break down proteins, aid in beginning digestion and
dissolve material into process able pieces. The next section will focus more closely on the
responsibilities and functions of each stomach compartment.
4. Small Intestine: The small intestine has three main sections the duodenum, jejunum and
ileum that work together to complete most of the actual digestive process. In the duodenum,
the section connected to the stomach, secretions from the gallbladder and pancreas mix with
the partially digested matter. This process balances the pH in the intestine, ensuring the
digestive enzymes work correctly. The jejunum section is lined with small, finger-like
projections known as villi, which increase the intestinal surface area and absorb nutrients.
The ileum absorbs vitamin B12, bile salts and any nutrients that passed through the jejunum.
At the end of the ileum is a valve, preventing any backward flow of materials. Throughout
the small intestine, muscular contractions move the matter forward. In a fully mature cow, the
entire organ may be up to 150 feet long and has a 20-gallon capacity.
5. Cecum: Sitting between the small and large intestines is a three-foot-long pouch called the
cecum. It has little function besides providing storage and a transition between the two
intestines, but it does aid in the continual breaking down of material. The cecum has about a
two-gallon holding capacity.
6. Large Intestine: Smaller in length but larger in diameter than the small intestine, the large
intestine is the final step of the digestive process. It absorbs remaining water and contains
bacteria microbes that finish digestion and produce vitamins the animal needs to grow and
remain healthy. Its last job is to eliminate any undigested and unabsorbed food from the
system in the form of waste. When the cow is properly handled and fed, this process
continually occurs, keeping the animal healthy and at the right weight. The entire digestion
process should take anywhere from one to three days. If something interrupts this process or
the cattle is unhealthy, the sections will no longer be able to function as well as they should,
causing diseases and complications.
The Four Components of a Cattle’s Stomach: Of the six components in the cattle’s
digestive system, the most important part is the stomach. A ruminant animal’s stomach has
four distinct compartments, each with its specific function. These compartments are:
1. Rumen: The rumen, also known as the “paunch,” is the first area of the cow’s stomach,
connected to the cattle’s esophagus. This compartment acts as storage for chewed vegetation
and forms balls of cud. Cud consists of large, non-digestible pieces of plant matter that must
be regurgitated, chewed a second time and swallowed before continuing through the process.
The rumen absorbs nutrients through papillae of the rumen wall and facilitates fermentation,
creating the rumen bacteria and rumen microbes necessary to break down and digest the
proteins in feed. Microorganisms in the rumen are responsible for digesting cellulose and
complex starches, as well as synthesizing protein, B vitamins and vitamin K. As a storage
area, it can hold up to 40 gallons of material. The rumen, combined with the reticulum, makes
up 84% of the volume of the entire stomach. A few common health issues with the rumen
include bloat, which occurs when a cow can’t eradicate a build-up of gas, acidosis and
rumenitis, which occur when low pH balance allows for high acid production. These can be
prevented by managing and paying attention to cattle’s food and water intake.
2. Reticulum: The reticulum is frequently referred to as the “honeycomb,” because the inner
lining appears like and is structured similar to a honeycomb. While it does have its
independent functionality, the reticulum is attached to the rumen with only a thin tissue
divider. This component holds heavy or dense objects such as metal pieces and rocks and trap
large feed particles that are not small enough to be digested. The reticulum facilitates
regurgitation. Both the rumen and reticulum contain digestive bacteria, so no acid is included
in the regurgitation of materials. The reticulum holds about 5 gallons of material. One
common health issue involving the reticulum is hardware disease, which occurs when cattle
ingest heavy or sharp objects like nails, screws or wire. They are swept into the reticulum and
may puncture the stomach wall. This disease is preventable by putting magnets on feeding
equipment to catch any metal, or cured by the placement of an intra-ruminal magnet that traps
already swallowed objects.
3. Omasum: The globe-shaped omasum is nicknamed “manyplies” because of its internal
structure. It is lined with large leaves and folds of tissue that resemble the pages of a book.
These folds absorb water and nutrients from feed that passes through after its second round of
chewing. The omasum is smaller than the rumen and reticulum, making up about 12% of the
stomach’s total volume. It can hold up to about 15 gallons of material.
4. Abomasum: The abomasum is the last component of the stomach and is often known as
the “true stomach,” because it operates the most similar to a non-ruminant stomach. This true
stomach is the only compartment of the stomach lined with glands. These glands release
hydrochloric acid and digestive enzymes to help the abomasum further break down feed and
plant material. In comparison to the other chambers, the abomasum is on the smaller side,
representing about 4% of the total stomach volume and only holding about 7 gallons of
material.
Each of these components is vital in maintaining a healthy digestive process. They must
cooperate quickly and efficiently to turn grain and plant matter into energy for the cattle. If
one section becomes incapable of performing or ceases to work correctly, it will affect all of
the other functions in the digestive system.
Because the rumen is the largest area of the stomach and the section that focuses on reducing
feed to be passed through the digestive process, it is crucial that it is properly developed and
remains healthy.
Carbohydrate Digestion and Absorption
The primary site of carbohydrate digestion is in the lumen of the small intestine, where
pancreatic amylase begins the digestion of starch granules (amylose and amylopectin). In
some birds, there is some salivary amylase action in the mouth, but not in farm animals.
There are two forms of amylase, one that cleaves α 1, 4 bonds in a random fashion,
while the other removes disaccharides units (maltose) from the polysaccharide chain.
Pancreatic amylase does not act on α 1, 6 bonds that form the branch points in the
structure of amylopectin.
The end products of amylase digestion include a mixture of glucose, maltose, and
dextrins (residues containing α 1, 6 branch points). Dextrins are acted upon by α 1, 6
glucosidase. The small intestine is the site of the digestion of carbohydrates in farm
animals.
Dietary simple sugars, such as glucose and fructose, do not need to be digested, as
they can be absorbed through the intestinal epithelium directly. The end products of
starch digestion diffuse into the brush border, where the final digestive processes
occur.
Disaccharides such as maltase and isomaltase on the intestinal brush border then
complete the degradation and are hydrolyzed to their constituent monosaccharides by
enzymes on the brush border, and the monosaccharides released are absorbed into the
enterocyte.
Sucrose is acted upon by sucrase to yield glucose and fructose for absorption. In
young animals kept on milk (pre-weaning), lactose is acted upon by lactase to yield
glucose and galactose. Amylase activity is very low in young animals consuming milk
 and is stimulated by solid food consumption. The end product of carbohydrate
digestion in monogastric animals is mainly glucose.
Monosaccharides are absorbed both by simple diffusion and adenosine triphosphate
(ATP)-dependent active transport. A sodium-dependent glucose transport protein
binds glucose and Na+ and transports them through the enterocyte and releases them
in the cytosol.
Carbohydrate-Digesting Enzymes
 Amylase
 Disaccharidase
 Maltase
 Sucrase
 Lactase
Overview of carbohydrate digestion: Site, enzymes, and end products produced in
monogastric animals.

Site Enzyme Product

Mouth Salivary Amylase Not too significant

Pancreatic Amylase
Small Intestine Monosaccharides
Di/oligosaccharidases

Large Intestine None Some volatile fatty acid

Monogastric animals do not secrete enzymes that digest the complex carbohydrates (β
1,4 linkages; e.g., nonstarch polysaccharides [NSP], glucans, cellulose) that are
components of plant fiber (e.g., wheat, barley) and are acted upon by hindgut
microbes to yield volatile fatty acids (VFAs).
High levels of NSP and glucans in a monogastric diet can cause viscous digesta and
can interfere with digestion processes leading to malabsorption. In poultry, high-NSP-
containing diets (e.g., barley, rye) can produce wet litter, dirty eggs, and diarrhea.
Carbohydrate Digestion in Ruminants: Carbohydrate digestion in ruminant animals is
through microbial fermentation in the rumen. Dietary carbohydrates are degraded (fermented)
by rumen microbes (bacteria, fungi, protozoa). The purpose of rumen fermentation is to
produce energy as ATP for the bacteria to use for protein synthesis and their own growth.
VFAs, also known as short-chain fatty acids (shown below), are produced as a product of
rumen fermentation and are absorbed through the rumen wall and are utilized by the animal
as an energy source.
Major Volatile Fatty Acids Produced in the Rumen
Acetic acid
Propionic acid
Butyric acid
The three major VFAs are acetic (C2), propionic (C3), and butyric acid (C4; shown below).
The end products of digestion in ruminants are volatile fatty acids and some
monosaccharides.
In young ruminants, rumen and the reticulum are not fully developed and are
relatively small. The reticular/esophageal groove reflex, a tube-like fold of tissue,
channels milk or water that is sucked from a nipple directly through the omasum to
the abomasum. This is a reflex, stimulated by sucking.
When the animal is weaned, it normally loses this reflex. Solid food, such as creep
feed, passes into the small rumen and fermentation starts. The neonatal ruminant
animal has no ruminal bacterial population but from birth, it starts to pick up bacteria
from the mother and environment, particularly through contact.
Solid food is then fermented forming VFAs, which stimulate the growth and
development of the rumen, particularly the growth of the papillae for absorption. All
the digested and absorbed monosaccharides and volatile fatty acids enter into the
liver.
The end products of rumen fermentation are microbial cell masses, or microbial
protein-synthesized VFA, and gases such as carbon dioxide, methane, hydrogen, and
hydrogen sulfide.
The products of fermentation will vary with the relative composition of the rumen
microflora. The microbial population also depends on the diet, since these changes the
substrates for fermentation and subsequently the products of fermentation. For
example, starch is the major dietary constituent in concentrate-fed ruminants (e.g.,
feedlot cattle). The rumen of such animals will have higher amylolytic bacteria than
cellulolytic bacteria present in the rumen of roughage- and pasture-fed animals.
Factors such as the forage:concentrate ratio, the physical form of the diet (ground vs.
 pelleted), feed additives, and animal species can affect the rumen fermentation
process and VFA production.
Molar ratios of VFAs are dependent on the forage:concentrate ratio of the diet.
Cellulolytic bacteria tend to produce more acetate, while amylolytic bacteria produce
more propionic acid. Typically three major VFA molar ratios are 65:25:10 with a
roughage diet and 50:40:10 with a concentrate-rich diet. Changes in VFA
concentration can lead to several disorders of carbohydrate digestion in ruminants.
Rumen acidosis occurs when animals are fed high-grain-rich diets or when animals
are suddenly changed from pasture- or range-fed to feedlot conditions.
Carbohydrates, Metabolism: Carbohydrates are the major source of energy in the animal’s
diet. By the help of different metabolic pathways by which absorbed glucose and volatile
fatty acids are converted to energy in the animal body.
Animals need energy to carry out all the body processes (e.g., nutrient transport, synthesis,
muscle contraction) required to maintain life. Without energy, an animal is unable to move,
to digest its food, to reproduce, to grow, or even to breathe. Energy requirement and balance
are more important in food-producing animals with their need to synthesize nutrients (e.g.,
proteins, fat) for deposition into muscle, milk, and eggs. Carbohydrates are the major energy
source in the diet of farm animals.
Forms of Energy
ATP is the compound used as an energy source in biochemical reactions.
Hydrogen plays a prominent role in energy metabolism. During the catabolism of glucose
(C6H12O6) by the animal, hydrogen is transferred from glucose to hydrogen receptors, such as
nicotinamide adenine dinucleotide (NAD+) and flavin adenine dinucleotide (FAD). These
hydrogen acceptors (reducing equivalents) are oxidized in the reactions of the respiratory
chain inside the mitochondria to release energy.
In biological systems, oxidation of hydrogen is coupled with the synthesis of adenosine
triphosphate (ATP). ATP is the readily available form of energy (“molecular energy currency
unit”) in the cell. ATP has three components: a nitrogenous base (adenine), the sugar ribose,
and the triphosphate (figure 5.1). Energy is stored within the PO 4 bonds, and the release of
each phosphate bond generates eight kcal of energy.
Forms of Energy
1 mole ATP = 8 kcal/mol
Reducing equivalents
1 mole NAD, NADH = 3 ATP
1 mole FAD, FADH = 2 ATP

Figure 5.1: ATP Structure

Absorbed Glucose and Metabolic Conversions in the Animal Body
Absorbed glucose could be utilized by the animal for several functions. The first
priority is the formation of glycogen in the liver, which is stored in muscle and
hepatic tissue. However, the body stores very little as glycogen (a starch-like
compound), and glycogen could be rapidly hydrolyzed back to glucose through a
process called glycogenolysis. This helps in maintaining blood glucose at a narrow
range in normal healthy animals.
The second priority is oxidation to form energy (a major function) and is described in
the next sections. Since glycogen storage is limited, excess glucose is converted to fat
and is stored in adipose tissue. This is accomplished by the breakdown of glucose to
pyruvate through a process called glycolysis, which is then available for fat synthesis.
Cells Derive Energy from Glucose: Metabolic Pathways
Cells use different steps to break down the absorbed glucose to carbon dioxide and water
through different enzymatic reactions. The catabolism of glucose occurs in two metabolic
pathways: glycolysis and the tricarboxylic acid (TCA; also called citric acid or Kreb’s) cycle.
Two major pathways of glucose catabolism are glycolysis and the TCA cycle.
Glycolysis: Enzymes for glycolysis are located in the cytosol of the cell, and glycolysis
occurs in this part of the cell. Glycolysis is the breakdown of 6 C glucose into two 3 C end
product pyruvates in aerobic metabolism and lactic acid in anaerobic metabolism. It is a
catabolic pathway involving oxidation and yields ATP and NADH (reduced NAD) energy.
Glycolysis is the pathway by which other sugars (e.g., fructose, galactose) are catabolized by
converting them to intermediates of glycolysis. Fructose can be converted to fructose-6-
phosphate by hexokinase. Galactose can enter glycolysis by being converted to galactose-1-
phosphate followed by conversion (ultimately) to glucose-1-phosphate and subsequently to
glucose-6-phosphate (G6P), which is a glycolysis intermediate.
Energy Production Process through Glycolysis: Glycolysis has two phases: an energy
investment phase requiring the input of ATP (preparatory phase) and an energy realization
phase (pay off) where ATP is made (Figure 5.2).
Cells that utilize glucose have an enzyme called hexokinases, which use ATP to
phosphorylate the glucose (attaches a phosphorus group) and changes it into G6P. At
this point, the cellular “machinery” can begin to process the glucose.
Briefly, in the first reaction of glycolysis, hexokinase catalyzes the transfer of
phosphate to glucose from ATP, forming glucose-6-phosphate. Thus this step uses
ATP, which provides the energy necessary for the reaction to proceed. Glucose-6-
phosphate is converted to fructose-6-phosphate and subsequently to fructose-1,6-
biphosphate, which is cleaved to dihydroxy acetone phosphate (DHAP) and
glyceraldehyde-3-phosphate (G3P).
During this process, an additional ATP is required to phosphorylate the intermediate
fructose-6-phosphate. Therefore, the “preparation” of glucose results in two
molecules of ATP being used for every glucose molecule processed.
During the payoff phase, G3P is further processed to produce pyruvate. During this
phase, one NADH and two ATP are produced during the intermediate steps. The
DHAP produced can be simply converted into G3P and processed in a similar manner
as the first G3P. Therefore, one glucose molecule will result in the production of two
NADH, four ATP, and two pyruvate molecules.
 Glycolysis: Net Gain of ATP
Input = 2 ATP
Produces = 4 ATP and 2 NADH
Net gain = 8 ATP (aerobic)
 Figure 5.2: Glycolysis pathway in cytosol
Glycolysis (Aerobic)
Input: = 1 glucose molecule
Requires = − 2 ATP (Activation)
Produces:+ 2 pyruvate molecules+ 4 ATP+ 2 NADH (= 6 ATP)
Total = 6 + 4 =1 0 ATP
Net gain =10 − 2 = 8 ATP
Glycolysis: Overall Functions
ATP Production: For each molecule of glucose, 2 ATP (preparatory phase) were
used and 2 NADH, 4 ATP, and 2 pyruvate molecules (payoff phase) were generated;
which equals a net production of 2 NADH, 2 ATP, and 2 pyruvate molecules, and the
net gain of ATP is 8 per mole of glucose.
Production of Other Intermediates: Glycolysis provides pyruvate for the TCA
cycle, amino acid synthesis through transamination, glucose-6-phosphate (glycogen
synthesis), nicotinamide adenine dinucleotide phosphate, (NADPH) (fatty acid
synthesis; triglyceride synthesis), and dihydroxyacetone phosphate for glycerol
synthesis (the backbone of fat).
Pyruvate in the Animal Body: It is important to discuss the fate of pyruvate generated
through glycolysis. Pyruvate has different fates, depending on the conditions of the animal
and the cell type.
Fates of Pyruvate
 Lactic acid production
 Acetyl CoA production
Lactic Acid Production 
When oxygen is present, there is plenty of NAD+, so aerobic cells convert pyruvate to
acetyl coenzyme A (CoA)   for oxidation in the citric acid cycle. When oxygen is
absent, NAD+ levels can go down, so to prevent that from happening, lactate
dehydrogenase uses NADH and pyruvate is converted to either lactate (animals) or
ethanol (bacteria/yeast).
Anaerobic conversion of NADH to NAD+ provides much less ATP energy to cells
than when oxygen is present. Anaerobic metabolism of glucose generates only two
ATP per glucose. Once oxygen is depleted for the cell, another system will convert
the lactic acid back to pyruvate and produce glucose. Anaerobic metabolism of
glucose produces two ATP per glucose molecule.
Acetyl CoA Production
Acetyl CoA Production: Acetyl CoA production occurs in the aerobic state and serves
as the main precursor for the TCA cycle, lipogenesis, and ketogenesis (during
negative balance).
Acetyl CoA is converted to ATP through different steps in the TCA cycle. During this
conversion, the enzyme pyruvate dehydrogenase and different B vitamin–containing
coenzymes (thiamine, riboflavin, niacin, pantothenic acid) function through a series of
condensation, isomerization, and dehydrogenation reactions and produces several
different intermediates that are used for fat or amino acid synthesis. Pyruvate
dehydrogenase links glycolysis with the TCA cycle by converting pyruvate into acetyl
CoA.
To generate more energy from the glucose molecule, further biochemical processes
occur within the animal body. These include the enzymatic step pyruvate
dehydrogenase (PDH), which connects glycolysis (cytosol) with the TCA cycle in the
mitochondria. During this step, 3 C pyruvate is converted to an active form of acetic
acid called acetyl CoA, and CO2 is produced.
 Pyruvic acid is decarboxylated and the 2 H ions are picked up by NAD+ and thus it
provides two mole of NADH for each mole of glucose (net = 6 ATP produced). This
enzymatic step needs coenzyme A and its activity is highly regulated by the
concentration of acetyl CoA, ATP, and NADH.

Through PDH, one mole of glucose produces 2 NADH or 6 ATP.

TCA, Citric Acid, or Kreb’s Cycle
Functions of the TCA Cycle
 Recover more chemical energy
 Provide metabolic intermediates (e.g., citrate, α-ketoglutarate, oxaloacetate).
TCA cycle, citric acid cycle, or Kreb’s cycle (named after Hans Krebs who discovered the
pathway in 1937) includes a series of enzyme-catalyzed chemical reactions that occur in the
matrix of the mitochondria. Several B vitamin–containing enzymes function as coenzymes in
the pathway (e.g., thiamine, riboflavin, niacin). The TCA cycle is the key part of aerobic
respiration in cells. The TCA cycle also serves as a source of precursors for storage forms of
fuels (lipids) and building blocks, such as amino acids, in the animal body.
The TCA cycle is the central metabolic hub of the cell and occurs in the mitochondria.
Prior to entrance into the cycle, pyruvic acid is converted to acetyl CoA through PDH
as described previously. Acetyl CoA (2 C) enters the cycle by combining with a 4 C
compound called oxaloacetate and forms a 6 C citric acid. This reaction “pulls” the
cycle forward. Citric acid undergoes a series (about 10) of enzyme-catalyzed
conversions producing different intermediates (e.g., α-ketoglutarate, succinate,
fumarate, malate; shown in Figure 5.3).
In many of these steps, high-energy electrons are released to NAD, and NAD
molecules also acquire H+ and become NADH. In one of the steps, FAD serves as the
electron acceptor, and it acquires H+ and becomes FADH2.
In one of the reactions, one ATP is also synthesized from one acetyl CoA. At the end
of the cycle, the final product is oxaloacetic acid, which is identical to the oxaloacetic
 acid that begins the cycle and will pick up another acetyl CoA to begin another turn of
the cycle.
Altogether, the TCA cycle produces (per one mole of glucose or two moles of pyruvic
acid), two ATP molecules, six NADH, and two FADH2. Both NADH and FADH are
used in the electron transport chain to generate ATP. Overall, a net gain of 24 ATP
per mole of glucose is obtained through the TCA cycle. One mole of glucose
produces 24 ATP through the TCA cycle.

Figure 5.3. TCA cycle in the mitochondria 

The TCA cycle is the final common pathway for oxidation of carbohydrates, fats, and
proteins that provides energy.
Overall, for every glucose molecule fully metabolized to CO2 and H2O, we receive
38 ATP. There are eight kcal of energy in every ATP high-energy phosphate bond.
Hence net recovery of energy is 38 × 8 = 304 kcal. The efficiency of converting
glucose bond energy into ATP high-energy P bond is therefore 304/674× 100 = 45%.
Altogether, one mole of glucose produces the equivalent of 38 ATP (8 + 6 + 24), or 304 kcal,
through the different steps explained.
The rate-limiting step in the TCA cycle is the combination of oxaloacetate and acetyl
CoA to produce citrate. Because of this, it is essential to have adequate quantities of
 oxaloacetate in order to produce ATP and maintain cell viability. Nutritional diseases
such as ketosis result from a deficiency in energy production and therefore require
oxaloacetate as precursors to “jump-start” the TCA cycle.
The TCA cycle also produces intermediates that serve as a precursor for the synthesis
of fatty acids and amino acids. For example, citrate can be used for fatty acid
synthesis, while oxaloacetate can be used for nonessential amino acids (glucogenic
amino acids or most of the nonessential amino acids, e.g., alanine, serine, cysteine,
glycine). During low glucose conditions, these amino acids can produce glucose and
α-ketoglutarate can be used for glutamic acid synthesis.
TCA Cycle Intermediates: Roles
Citrate for fatty acid synthesis
α-ketoglutarate for glutamic acid (amino acid) synthesis
Oxaloacetate for non essential amino acid synthesis 
Electron Transport System, Respiratory Chain, or Oxidative Phosphorylation: The
electron transport chain permits recovery of redox energy associated with NADH and FADH.
H ions react with O2 and form water, and thus the electron transport system serves as
a source of metabolic water. Electrons are carried to the electron transport system in
the mitochondrial wall by NADH and FADH2.
Coenzyme Q accepts a pair of electrons and passes electrons singly to cytochrome C
and acts as a “traffic cop” for electrons. Oxygen is the terminal electron acceptor and
if oxygen is not available, electrons will not pass through the electron transport
system and NADH and FADH2 will not be reoxidized.
For these reasons, the citric acid cycle will not run either. This is part of metabolic
control. Interruption of electron flow can result in production of reactive oxygen
species (free radicals). Cellular enzymes, such as superoxide dismutase and catalase,
help deactivate reactive oxygen species.
Ruminant Carbohydrate Metabolism: Volatile fatty acids (VFAs) are produced in large
amounts through ruminal fermentation and are of great importance in that they provide
greater than 70% of the ruminant’s energy supply. The rumen epithelium performs efficient
absorption of VFAs through diffusion through a concentration gradient. As they pass through
the epithelium, the different VFAs undergo different degrees of metabolism.
The Fate of Volatile Fatty Acids
Acetate—precursor to Acetyl CoA (TCA cycle; fat synthesis)
Propionate—precursor to glucose (glycolysis; milk lactose)
Butyrate—precursor to Acetyl CoA (TCA cycle)
Acetate and propionate pass through the epithelium largely unchanged, but almost all the
butyric acid is metabolized in the epithelium to beta-hydroxybutyric acid, a type of ketone
body. The three major VFAs (acetic, propionic, butyric) absorbed from the rumen have
somewhat distinctive metabolic fates:
Acetic acid is utilized minimally in the liver and is oxidized throughout most of the
body to generate ATP. Another important use of acetate is as the major source of
acetyl CoA, and it enters the TCA cycle. Acetic acid is used for the synthesis of lipids
(e.g., milk or body fat). A high-roughage diet favors the production of acetic acid.
Propionic acid is almost completely removed from portal blood by the liver.
Propionate is converted to succinyl CoA, and it enters the TCA cycle. Within the
liver, propionate serves as a major substrate for gluconeogenesis, which is absolutely
critical to the ruminant because almost no glucose reaches the small intestine for
absorption. For example, in a dairy cow, all the glucose in the milk lactose was
synthesized in the liver and most of that synthesis was from propionic acid. A high-
concentrate diet favors the production of propionic acid.
For butyric acid, butyrate is split into two acetyl CoAs, and it enters the TCA cycle.
Most of the butyric acid that comes out of the rumen as the ketone beta-
hydroxybutyric acid which oxidized in many tissues for energy production.