Protein and Amino Acids

 Although all of the proteins in a feedstuff are often referred to collectively as proteins,
they do differ in individual composition.
 It is the specific sequence and composition of amino acids and the manner the amino
acids strands are connected to each other, that determine the physical and chemical
properties of individual proteins.

Functions

 Proteins are important structural parts of soft tissues like muscle, CT, Collagen, skin,
feathers, toe nails, beak etc.
 Blood proteins, albumin, globulin help to maintain homeostasis, regulate osmotic
pressure, and acts as reserve supply of aas.
 Fibrinogen and thromboplastin involved in blood clotting.
 Conjugated proteins like Hb carries O2, Lipo proteins transport fat soluble vitamins and
other fatty metabolites, constituents of cell membrane.
 Nucleo proteins in DNA, RNA, glycol proteins, Phosphoprotein (egg yolk vitellin),
Enzymes, hormones

Classification of Proteins

 Globular Proteins- Albumin, Globulin, Glutenin, Prolamines

 Fibrous Proteins- Collagen, elastin, keratin
 Conjugated proteins- Nucleo proteins, muco proteins, Glyco proteins, Lipo proteins

The protein of the plants varies with the type of seeds (cereals, oil seeds), maturity of the
leaves and within the seeds also.

 The proteins of cereals are largely Prolamines and glutelins whereas in oil seeds it is
mostly globulin known as glycinin.
 Among the species protein requirement of fish is higher than other (40%)
 Can’t utilize the carbohydrates (less amylase)
 Less energy expenditure due to NH3 excretion
 Energy for body temperature regulation is less (cold blooded animal)
Classification of Amino acids

 Amino acids which can’t be synthesized by animals or not adequately synthesized to

meet the requirement of the animals are classified as Essential or indispensable aas.
These aas to be supplied through the diet.
 Out of the essential aas few are present in limited amount in most of the protein
sources are referred as limiting aas.
 Those aas that can be synthesized by the animals are termed as non-essential or
dispensable aas.
 Out of the non essential aas few of them can’t be synthesized at a rate fast enough for
maximum growth which are referred as semi essential aas.

Nutritional classification of Amino acids

Essential aas Limiting aas Semi essential aas Non essential aas

Arginine Lysine Tyrosine Alanine

Lysine Methionine Cystine Aspartic acid

Histidine Hydroxy lysine Aspargine

Leucine Glutamic acid

Isoleucine Glutamine

Valine Hydroxy proline

Methionine Glycine*

Threonine Serine

Tryptophan Proline

Phenyl alanine

*Requirement of Glycine is essential during chick period.

The essential aas requirement for pig, Fish, hen = 10
The essential aas requirement for chicks = 11 (+ Glycine)
Human being =8 (-Arginine, Histidine)
 Rat- 12 ( + Aspargine,Proline)
All the aas occurring in animal tissues are L- isomers exception is Methionine (Both D
or L methionine).

Uric Acid synthesis

 Unlike mammals birds secrete waste or excess nitrogen as uric acid rather than urea
 The final step of uric acid synthesis is controlled by a molybdenum containing enzyme
Xanthine Oxidase
 Uric acid is extremely insoluble and when the blood levels exceeds (4-10 mg/dl) , uric
acid precipitates in joints, under skin, in kidney producing severe gout.
 Glycine is an integral part of uric acid molecule.
 Each time a molecule of Uric acid is excreted one molecule of Glycine is lost.
 That’s why Glycine requirement of chicken is very high. (1.5% of diet vs. no
requirement in other species
 Though Glycine id readily synthesized by chicken but during rapid growth and during
chick stage it is required.

Endogenous N2 and amino acid losses

 When adult birds are fed a protein free diet , the endogenous urinary nitrogen loss is
around 140 mg/kg 0.75 /day.
 Metabolic fecal losses are about 40 mg/kg 0.75 /day depending on the fibre content of
diet.
 So maintenance endogenous nitrogen losses total about 180 mg/Kg 0.75 /day.
 It is estimated that total endogenous nitrogen losses in growing chicken range from
200-300 mg/kg body weight/day.

Energy cost of protein Synthesis

 Both the synthesis and degradation of protein and amino acids are energy demanding
process.
 Uric acid synthesis is much more energy demanding than urea synthesis.
 That’s why birds inherit an energy cost for excreting an insoluble nitrogenous product.
 Energy cost of uric acid synthesis is about 330 Kcal/mole compared to just 85 K
cal/mole of urea production. (3.88 times)
 Energy cost of protein deposition to be around 0.7 K cal/g protein deposition
 Buttery et al., (1973) calculate the total costs of protein anabolism, turn over and uric
acid synthesis represents about 11 % of ME intake.

Protein and amino acid Nutrition of Growing birds (Pullets)

 We can model the protein requirement of pullets based on needs for Maintenance,
Growth and Feather development.
 Total Protein Requirement= Protein for maintenance (P m) + Protein for Growth (P g) +
Protein for Feather Development (P fd).
 Protein Requirement for Maintenance (Pm)= Daily Endogenous Nitrogen loss in
younger chicken has been determined to be approximately 250 mg N2/kg body weight.
So Protein lost /kg body weight = 250 x 6.25= 1600 mg/kg= 1.6 g/kg
The efficiency of protein utilization is 61 %. So the dietary protein requirement is
1.6/0.61= 2.6 g/kg body weight
 Protein Requirement for Growth (Pg)- The carcass of chicken contains approximately
18 % protein.
So daily protein requirement for growth is calculated by multiplying the daily body
weight gain (in g) by 0.18 and dividing 0.61 (61 % efficiency of utilization).
Suppose the daily gain is 10 g , the Pg= 10 x 0.18/0.61=2.95 g/day
 Protein Requirement for feather development (P fd)- At 3 weeks of age the feather
represents about 4% of body weight. And at 4th week it is 7% which remains relatively
constant thereafter.
The feather contains about 82 % protein.
Daily protein requirement for feather development can be calculated by multiplying
the % feather weight (0.04 -0.07) times the daily gain in body weight in gram and
multiplying with 0.82 and dividing with 0.61.

 The total protein Requirement for growing WLH chicken = Pm + Pg + Pfd

0.0016 x body weight (g)/0.61 + Daily gain (g) x 0.18/0.61 + 0.07 x daily gain (g) x
0.82/0.61
  Protein requirement for Egg production – Egg contains around 12 % protein (54 g x
0.12=6.48 g)
Let the egg production level is 80 % , so 6.48 x 0.80= 5.2 g
5.2/0.60 (efficiency of utilization)= 8.66 g

Example- Lets the body weight of a WLH layer bird is 1.5 kg , the egg production level of the
flock is 90% and the genetic potency of body weight gain of the strain from 24 to 40 weeks is
450 g. Calculate the protein requirement of these layer birds.

Ans.:-

The total protein Requirement= Protein Requirement for Maintenance (Pm) +

Protein Requirement for Egg Production (Pegg) +

Protein Requirement for Growth (Pg) +

Protein Requirement for Feather Development (Pfd)

Pm= Daily Endogenous N loss is 250 mg/kg body weight or 1.6 g protein/kg body weight

1.6 g x 1.5 kg body weight= 2.4 g protein/d

Taking efficiency of utilization to be 60 %-The Protein Requirement is 2.4/0.6=4 g/d

Pegg = An standard egg weighs 54 g and contains 12% protein so an egg contains 6.5 g protein

Egg production level of the flock is 90%, so the requirement is 6.5 x 0.9= 5.85 g

Taking the efficiency of protein utilization 60%, the requirement is 5.85/0.6=9.75 g

P g = The strain growth potency is 450 g in 16 weeks i.e. 450 g/112 d = 4 g/ d

The protein requirement for growth is = (Daily gain (g) x 0.18)/0.6= 4 x 0.18 /0.6=1.2 g

(Carcass contain 18 % protein and efficiency of protein utilization 60%)

P fd= Feather represents 7% of body weight and protein content of feathers is 82%

Protein requirement for feather development= (Daily gain (g) x 0.07 x 0.82)/0.60=0.38

Ans.

Total protein requirement= Pm + Pegg + Pg + Pfd= 4+9.75 +1.2 + 0.38=15.33 g/day

Model Predicted for Amino Acid Requirement

 Numerous mathematical models have been developed that aimed at predicting the
amino acids needs of laying and growing birds. For laying hens these include
maintenance, growth and egg production.
 Combs (1960) was one of the first scientist to predict aa requirements of laying hens
Amino acid (mg/bird/day) = a W + b ∆W + c E
Where W= body weight
∆W= Growth
E= Egg mass
a, b, c are constants
 Mc Donald & Morris (1985) used another model to predict three important aa needs
of young pullets in mg/bird/day

Lysine= 9.99 E + 73 W

Methionine = 4.77 E + 31 W

Tryptophan = 2.62 E + 11 W

Where E= daily egg mass (g)

W= Body weight (kg)

Example- Let the body weight of the hen is 1.5 kg and producing an egg of 56 g the
important aas requirement as calculated using these formulae

Lysine= 670 mg/d, Methionine= 314 mg/d and Tryptophan= 164 mg/d

 Knowing the amino acid requirement of poultry and the aa content of feed stuffs, it
would seem to be relatively simple matter to
i) Calculate the aa composition of a feed
ii) Compare this with known aa requirement of animal at different stages as
recommended by the feeding standard (BIS, 2007)
iii) If necessary supplement with synthetic aa or with other protein sources
Requirement of nutrients and amino acids in chicken feed in different stages (BIS,2007)

Characteristic Broiler Feed Layer Feed

Pre Starter Finisher Chick Grower Layer Layer
starter (8-21 d) (>22nd d) (0-8 (9-20 phase-I phase-II
(1-7 d) wks) wks) (21-45 46-72
wks) wks)
Crude Protein (%) 23 22 20 20 16 18 16
min
ME (K cal/kg) 3000 3100 3200 2800 2500 2600 2400
min
Ether extract (%) 3.0 3.5 4.0 2.0 2.0 2.0 2.0
min
Calcium % 1.0 1.0 1.0 1.0 1.0 3.0 3.5
min
Total Phos.s % 0.70 0.70 0.70 0.65 0.65 0.65 0.65
min

Available Phos. % 0.45 0.45 0.45 0.45 0.40 0.40 0.40

min
Linoleic acid % 1.1 1.1 1.1 1.0 1.0 1.0 1.0
Min
Lysine % min 1.3 1.2 1.0 1.0 0.7 0.7 0.65
Methionine % min 0.50 0.50 0.45 0.40 0.35 0.35 0.30
Methionine + 0.90 0.90 0.85 0.70 0.60 0.60 0.55
cystine % min
Moisture % max.
11 11 11 11 11 11 11
Crude fibre % max 5 5 5 7 9 9 10
AIA % max 2.5 2.5 2.5 4.0 4.0 4.0 4.5
Common Salt % 0.5 0.5 0.5 0.5 0.5 0.5 0.5
max
Aflatoxin B1 (ppb) 20 20 20 20 20 20 20
max
 Protein requirements of Layers

Many factors influence feed consumption and protein requirement of mature laying hens.
These are

 Size and breed of the hen

 Environmental temperature
 Daily egg mass
 Stage of production
 Housing (cage/floor)
 Feeding space, stocking density
 Disease status
 Availability of drinking water
 Energy content of the diet
 Size and breed of the hen: - Heavy breeds of chicken consume more feed than light breeds
and they require more maintenance energy and more protein per day for maintenance. (as
endogenous N loss depends on body weight).
Generally coloured breeds (RIR, Vanaraja, Red Cornish) are heavier than white one (WLH, BV
400), and consumes 115-125 g of feed @ 18% protein that amounts to 21-23 g protein daily
whereas a WLH layer under similar environmental condition consume 100 g of feed daily with
18% protein and intake of protein per day may be 18 g .
 Environmental Temperature: - With conventional corn soyabean based diet having 2800 K
cal/kg the WLH hens consume 110 g feed/d during winter months (13 0c) but the birds will
consume 90 g feed during summer (30 0 c).
Thus in minter a protein level of 16% would provide 17.6 g protein daily while in summer a
protein level of 19.5 % is required to provide the same.
In practice the protein level of the diet may not be increased as this enhances the SDA (heat
increment) of the diet causing more heat stress. It is important to maintain the intake of
limiting/ critical amino acids like methionine and lysine in the diet which may be met from
synthetic source.
  Stage of Production:- Egg production commences at 18-20 weeks of age and reaches
peak production (95%) about 24-28 weeks, maintains upto 40-45 weeks of age then
gradually declines to a level of 75% after 60 weeks of laying (72 weeks of age)
The production cycle may be conveniently divided into 2 phases.
 Phase- I- Onset of lay (18-20 wks) the average WLH pullet weighs 1350 g and
consuming 75 g of feed daily.
During the period from 19-42 weeks of age, the pullet is expected
i) To increase egg production from 0-95%
ii) Increase body weight from 1350- 1650 g
iii) Produce eggs of gradually increasing size from 43-61 g

This phase is most critical period in the productive life of pullet. This necessity for providing
adequate protein, amino acids , vitamins, minerals for optimum egg mass, normal growth to
maturity and maximum economic return.
It also necessary to provide mature hen with health and tissue reserve needed for maximum
production throughout the subsequent stages of her productive life.
 Phase-II- It is the period after 42 weeks of age when the birds have attained mature
body weight (42-72 weeks) . Egg production declines more slowly at this time and there
is a minimal weight gain and increase of the egg size.

Daily protein needs of WLH hens during 2 phases of Egg production

Protein needed (g/d)

Phase-I Phase-II
1.Production of an egg 12.2 13.5
2.Maintenance of body 3.0 3.4
protein for one day
3.Growth per day 1.4 0
4.Feather growth/d 0.4 0.1
Total 17.0 17.0

The average egg size during phase I is 54-56 g and in phase -II it is 60-62 g and the
protein content is 6.7 g and 7.4 g respectively.
 Fresh Egg contains

Water 66 %
Protein 12 %
Fat 10 %
Carbohydrate 1%
Ash 11 %

 Protein and Egg size- Egg size is greatly influenced by genetics and body size of the bird
.
 At normal level intake Energy level has no effect on egg size.
 Protein and amino acid especially Methionine have a large influence on
egg size
 In a study by Calderon and Jensen (1990 & 1995) showed a generalized
curvilinear response in egg weight to graded levels of methionine (by
feeding 0.255 to 0.440 % methionine the egg size increased from 56 to
60 g , around 5-7 % increase of the egg weight.
 Ronald et al., (1998) showed a consistent linear trend for increase in
the egg weight of young birds (25-33 wks) as the TSAA was increased
from 0.65 to 0.81 % ( 49.3 vs. 51. 6g in 25 wks an55.3 g vs. 56.3 g in 33
wks)
 The other nutrients most often considered while attempting to
maximize egg size is Linoleic acid. In most situation 1 % dietary linoleic
acid meet birds demad. Although for mamimizing egg size as high as 2
% are often used.
 As a role of methyl donor, Choline also influences the egg size. As
choline can spare some methionine in the diet, because from
homocysteine methionine can be formed by addition of methyl group
from choline. Similarly choline can be formed from ethanolamine taking
methyl group from methionine. But the second one is not desirable as
methionine is very scarce in the diet and also not economical to add
synthetic methionine.