Seminar on

Protein Degradability Concept of Protein

Evaluation in Ruminants.

By Mayank Tandon

Dairy Cattle Nutrition Division, NDRI, Karnal

email;

mayanktandon1@gmail.com

INTRODUCTION

India Ranks Ist in Milk Production 91 MT/ annum for 2004-05 (NDDB,2004)

Bovines in India Cattle 186 million Buffaloes 97 million (Livestock Census, 2003)

Increasing @ 1% annually.Shortage of 30-32 % of Feed

Shortage Exist both for Energy and Protein Feeds.

Good Quality Protein Feeds are Expensive Lower area under oil seed production Export of oilseed Adequate Supply of Protein is Crucial to the Performance of Dairy Animals. A Protein Deficiency/ Imbalance of Amino Acids (AA) have a Dramatic Effect on Growth Milk production And other functions (Walli et al., 2004)

CONCEPTUAL CHANGES IN PROTEIN SYSTEMS A. Digestible Crude Protein System(DCP) Starting one, Easy to use, Widely accepted & used till date

“The DCP content of feedstuffs is measure of the N x 6.25 that has apparently disappeared in the digestive tract.” (ARC, 1965; NRC, 1970)

B. Metabolizable Protein System (MP) In USA, (Burroughs, et al.,1971) “As the quantity of protein digested or absorbed in the post ruminal portion of the digestive trace of ruminants.”

Comparison of Metabolizable Protein (MP) system with NRC & ARC Digestible Protein ( DCP) systems.

AnimWt.,

Kg

NRC, 1970 ARC, 1965 Metabolizible protein (Burroughs, et al.,1971)

Required, gm

observed

gm

difference

R, gm Observed

D R, gm O Difference

150-200 355 453 -22% 348 453 -23% 486 468 +4%

200-250 448 471 -5% 383 471 -19% 469 478 -2%

213-418 555 476 +17% 447 476 -6% 461 444 +4%

213-472 519 638 -20% 512 638 -20% 542 584 -7%

224-451 604 565 +7% 435 565 -23% 433 470 -8%

C. Absorbed True Protein (AP) System

In America, very much similar to

Metabolizible Protein system.

D. Proteins Digestible in Intestine (PDI) System

In France, to replace DCP (Verite et al., 1979)

“It estimate the quantity of amino N x 6.25 absorbed

in the small intestine from the dietary proteins which

has escaped fermentation in the rumen and the

microbial protein arising from that fermentation.”

Comparison of PDI and DCP Requirements. (Verite et al., 1979)

Type of animal

Wt.

kg

Level of production

Requirements (g/d)

PDI DCP

Fattening

Young Bull 400

Dairy, 1.2 kg/d 635 645

Beef, 1.4 kg/d 720 740

Dairy cows

600

maintenance 395 360

Last month of pregnancy

600 600

Lact, 30 kgFCM 1895 2160

E. Digestible Protein in the Intestine System

The Dutch system; Very much similar to PDI system “Quantity of Protein Digested in the Intestine.”

F. Crude Protein flow at the Duodenum The German (Kaufmann,1979) N x 6.25 flow at the duodenum; provided by Microbial Protein + Escape Protein G. ADPLS system Apparently Digestible Protein Leaving Stomach In Australia

RDP and UDP System (Roy et al., 1977; ARC, 1980 & 1984; NRC, 1989) Ruminants have protein requirement at 2 levels I. The N needs of rumen microbes. II. The protein need of the Host. Dietary protein have 2 parts RDP & UDP RDP Rumen Degradable Protein, part of the feed protein which degraded in rumen

Protein Degrading Microbes

(Yokoyama and Johnson, 1988; Lockiuood et al., 1988; Asao et al ., 1993)

Enzymes for Protein Degradation in Rumen

Trypsin like proteinase, Cysteine proteinase, Serine proteinase Metallo proteinase, Exopeptidase, Aminopeptidase, Deaminase etc.

Bacteria Protozoa Fungi

Bacteroides amylophilus

Butyrivibrio fibrisolvens

Streptococcus bovis

Bacteroides ruminicola

Holotrics

Isotricha sp.

Dasytricha sp.

Niocallimastix frontalis

Piromsces sp.

Orpinomyces joyonii

UDP Undegradable Dietary Protein

“Which Escape(Bypass) the Rumen Fermentation gets digested in the SI to supply AA.” Later also called as Naturally Protected Proteins Its Depends Upon surface area available for microbial attack chemical nature of proteins physical consistency of proteins other dietary components passage rate from rumen

Determination of Protein Degradability In vivo, using flow rates (cannulated) In Situ Nylon Bag Technique (Mehrez and Orskov, 1977; Orskov and Mcdonald, 1979)

Limitation Bag size, Pore size, Sample size, Particle size, Method of washing, etc. In vitro single stage technique double stage technique (Tilley and Terry, 1963)

Percentage of UDP in some Common Feed and Fodder

( NRC, 1985; Dutta et al., 1997; Negi et al., 1989)Feed UDP % Feed UDP %

Maize (grain) 65 Blood meal 76 – 82

Barley 21( 11-27) Fish meal 71 – 80

Sorghum 52 Meat meal 53 – 76

Bajra 68 Brewers dried 53

Oat grain 14–20 Corn gluten 53

Wheat grain 20–36 Wheat bread 29

Cotton seed meal 41–50 Corn silage 27

Linseed meal 11–45 Rice straw 63

Ground nut meal 30 Wheat straw 45

Rapeseed meal 23 Para grass, fresh 52

Soybean meal 28 ( 15–45) Cow pea 32 – 45

Sunflower meal 24 Berseem 37 – 52

Subabul 51 – 70 Alfa-Alfa 28

Nutrient Requirement for Lactating Cow of 400 kg B.Wt. Producing Milk with 4% Fat (NRC, 1989)

Milk Yield, kg

(4%fat)

ME

(MJ/d)

CP

(g/d)

CP provided by Microbes

(8.34xME, (MJ/d))

UDP required

(CP required – RDP)

0 50.44 318 420.66 Nil

1 55.65 408 464.12 Nil

2 60.86 498 507.57 Nil

3 66.07 588 551.02 36.98

4 71.27 678 594.47 83.61

5 76.48 768 637.84 130.16

6 81.7 858 681.37 176.63

7 86.91 948 724.82 223.18

8 92.12 1038 768.27 269.73

9 97.12 1128 811.72 316.28

10 102.54 1218 855.17 362.83

In general,

Requirement for RDP = 60-65% of CP

Requirement for UDP = 35-40% of CP

( NRC, 1989)

Various Treatments to Increase UDP Content in the Feed Stuffs.

Treatments Method Reference Comment

Heat 125-1500 C

For 30-45 min

Chalmers et al., 1954 Bartly & Deyoe,1975

Much of work

Under/ over protection

HCHO 1-1.2 g HCHO/ 100g CP

Ferguson et al., 1967

Chatterjee & Walli, 1997

Most studied

Cost effective and widely accepted; likely to be phase out

Esophageal

Groove Closer

Normal function in young; for liquid Proteins;

Salts of Na, Cu, Ag & Zn can effect

Orskov & Fraser, 1969

Orskov et al., 1970

Not Practicable

Cont…….

Post Ruminal Infusion (fistula)

Surgically Fitted

……..

…….. Only for Research,

Not Practicable

Encapsulation

of proteins

Gelatin capsules

Tristearin coat etc.

Strom & Orskove, 1984

For good BV proteins & AA; Methionin & Lysine, cost

Amino Acid

Analogs

Structural altering of AA,

Methionin hydroxy

N-acetyl-Dl-Methionin

DL-Homocysteine

DL-Homocysteine thiolactone-HCL, etc.

Amos et al., 1974

For Individual AA

Availability at SI

Cost

Feed Processing

Grinding, Pelleting, etc. (Heat)

Disruptution of protein matrix

……

…….

Can either Increase or Decrease UDP

Cont…

Ruminal Protease Activity

Antibiotics can be used Hogan & weston, 1969

Banned, adverse effect on fibre degradability,

Metal AA

Complex

Zn-Methionin,

Zn-Lysine, Cu-Lysine

Mn-Methionin

Fe-Methionin

Dass, 2003 For individual AA;

Better

Plant Secondary compounds

Lignin, Tannin, Terpeniods, Volatile essential oils, Alkaloids etc.

……

……

Have potential to be used;

Toxic effect

Retention time in Rumen

Less stay less egradation, Feed intake, specific gravity, partical size, concentrate : roughage, salt, water, tempt.

….

.. ….

Development of CNCPS system

AA Requirements Addressed (O'Connor et al., 1993)

Conceptual Improvement over RDP/UDP system

AA available by Microbial Protein + UDP

Feed Proteins have 3 Fractions (Chalupa & Sniffen, 1996) A ( Non Protein N) B (True Protein) C (Bound protein)

B is further Fractionated B1 (Readily Degraded in Rumen) B2 (Slowly Degraded in Rumen) B3( Hardly Degrade in Rumen )

Composition, Ruminal Degradation and Intestinal Digestion of Protein Fractions as per CNCPS (Chalupa & Sniffen, 1996)

Fraction Composition Rumen

Degradability %

Intestinal Digestibility %

A NH3, NO2, AA & Peptides Instantaneous Not reach

B1 Globulines, some Albumins Highly degradable 100

B2 Most Albumins, Glutelins 5-10 100

B3 Prolamins, denaturated &extended Proteins

0.1-1.5 80

C N bound to lignin,Mmillard Proteins

Zero Zero

Chemical Composition

Full Fraction

PBSN Phosphate Buffer Soluble N

A + B1

PBIN Phosphate Buffer Insoluble N

100 - PBSN

NDIN Neutral Detergent Insoluble N

…..

ADIN Acid Detergent Insoluble N

C

PBIN - NDIN …… B2

NDIN – ADIN …….. B3

Composition According to Borate-phosphate Buffer as per CNCPS, (Chalupa & Sniffen, 1996)

Ingredients CP PD

%

PBSN (A+B1)

PBIN-NDIN (B2)

ADIN-NDIN (B3)

ADIN

( C)

Mustard Cake 331.8 85.6 76.2 10.5 9.9 3.2

Ground nut Cake

731.9 84.6 74.1 5.3 15.3 5.1

Barley 102.5 88.3 51.3 12.1 32.3 4.1

Wheat Bran 135.6 86.2 46,5 30.1 30.3 3.0

Wheat 89.4 85.3 37.8 30.4 27.5 4.1

Protein fraction of some feeds Asper CNCPS(Mondal & Walli, 2003)

Ingredients CP

G/kg

PD

%

PBSN (A+B1)

PBIN-NDIN (B2)

ADIN-NDIN (B3)

ADIN

( C)

Deoiled Coconut Cake

255.0 83.2 43.8 12.9 38.1 5.0

Sunflower Meal 363.1 67.6 37.7 25.3 31.1 5.8

Soybean Meal 548.1 80.2 41.6 15.9 35.7 6.7

Fish Meal 490.6 81.0 27.8 24.3 44.3 3.5

Maize Grain 125.6 76.5 23.7 57.5 14.7 3.9

Cotton Seed Cake 291.9 78.5 32.1 20.0 41.4 6.3

Maize Gluten Meal

703.1 79.2 11.6 33.4 46.1 8.7

Conti…..

INDIAN CONTEXT

Indian Dairy cattle & Buffalo have

Lower Basel Metabolic rates then Temperate

Lower maintain requirements

Low producing animals.

NRC & ARC standards; for Animals raised;

Temperate Condition with High Quality Feeds.

Indian ---Tropical--- Low Producing---Poor Quality Feed

In India DCP & TDN System is More Popular

Simple, & Values are Available

Maintenance Requirements of Adult Cattle & Buffaloes ( ICAR, 2002)

Bt. Wt.

(kg)

DCP

(g)

TDN

(kg)

ME

(Mcal)

200 150 1.7 6.0

300 200 2.4 8.4

400 250 3.0 10.8

500 300 3.7 13.2

Requirements of Milch Animals Over and Above the Maintains Allowance ( per kg or liter of Milk)

(ICAR, 2002)

% Fat in Milk DCP (g) TDN ( kg) ME (M cal)

3 40 0.27 0.97

4 45 0.315 1.13

5 51 0.370 1.28

6 57 0.410 1.36

7 63 0.460 1.54

8 69 0.510 1.80

9 75 0.550 2.06

10 81 0.600 2.16

In last Decade,

Lots of work has been done on RDP/UDP

Including Various Treatment to Increase UDP Values

Feed Industry has Adopted Heat & HCHO Treatment

Recently Work is going on CNCPS system

Protein degradability Concept

At Present Most in Use

DCP; RDP/UDP & CNCPS

RDP/ UDP system is Mostly Followed World Wide ( NRC, 1989)

Response of Dairy Animals to RDP/UDP System Feeding

Or

Work Done by Various Scientist on Degradability Concept

Effects of RDP on ruminal fermentation

pH: no change or decrease

Ammonia : reduces

Total free AA: reduces

TVFA/ IVFA : no change (Hristov et al., 2004)

Effect of feeding concentrate mixtures with Varying levels of RDP to UDP on the yield and composition of milk in crossbred cows ( Kabande & Thomas, 1999)

GNC, Gingelly C, CSC, Coconut C, Maize, Wheat Bran, MM, Salt; Grass were Fed Adlib

Particular Concentrate Mixtures

A B C

Animals 6 6 6

RDP : UDP 37 : 63 52 : 48 70 : 30

Milk yield (kg/d) 10.1 7.18 6.32

Fat ( %) 4.68 4.63 4.64

Protein ( %) 3.63 3.48 3.46

Total solids ( %) 13.55 13.46 13.41

Effect of graded level of UDP feeding on milk Yield and composition (Chaturvedi & Walli, 2001)

aAttributes Diets T1 T2 T3

Cows 5 5 5

RDP : UDP 71 : 29 58 : 42 44 : 56

DMI ( kg/d) 11.54 10.66 11.18

Milk Yield ( kg/d) 9.81 9.87 10.09

4%, FCM (kg/d) 9.68 9.81 10.47

Fat (% ) 3.93 4.01 4.27

Protein ( %) 3.48 3.5 3.57

SNF ( %) 8.91 8.96 8.99

Total Solids ( %) 12.83 12.96 13.19

Wt. 460 kg; Maize fodder & wheat straw Barley,GNC, CSC, MGM, WB, MM, Salt

Performance of milk yielding cows on different degradable diets (Aharoni et al., 1993)

Parameters H D L D

Cows, no. 40 45

CP (%) TMR 16.7 16.9

CP degradability ( %) 69 64

Initial Wt (kg) 546 564

BW change ( kg/d) 0.178 0.146

Milk Yield ( kg/d) 31.8 33.4

Protein ( %) 3.04 3.06

Fat (%) 3.32 3.66

Effect of High & Low RUP Diets on Milk Yield & Composition in Dairy Cows ( Dunlap et al., 2000)

Attributes Dietary Treatments

High RUP Low RUP

Cows , no. 30 30

CP % of Diet 16 16

RUP ( % of CP) 35.4 28.6

DMI (kg/d) 21.3 21.4

Milk yield kg/d 32.6 32.2

3.5 % FCM (kg/d) 33.9 33.8

Fat ( %) 4.31 4.39

Protein ( %) 3.35 3.34

Milk yield, fat & protein % on feeding 1 kg bypassprotein supplement ( Garg et al., 2002)

Parameters Control Experiment

Cows, No 8 8

Milk yield, kg 14.1 15.2

Fat, % 4.4 4.6

Protein, % 3.2 3.5

Maize Fodder, Paddy Straw & Oat Silage1 kg procted ( HCHO) Sunflower Meal,

CP 28%, UDP 74% of CP

Nutritional facts for RDP/UDP

Good Utilization of Microbial Protein Synthesis

Additional Supply of AA/EAA at SI

Low Ammonia Production in Rumen

Low Urea Synthesis in Liver

Energy Savings

Excess AA go for Gluconeogenesis

Some Researchers have Reported Increased Milk Productions when Fed Higher Levels of UDP while others have not Observed any Significant Increase in Milk Yield (Sampath et al., 2003)

Naturally Protected Protein are often reported to have Methionine & Lysine as Limiting A A

Indian condition really need UDP ( Bypass proteins) ?!

Maintenance Low producing Animals

Mid Producer – Naturally Protected High producing – treatments

Methionine & Lysine – Microbial Protein (Strom & Orskove, 1989) - Naturally protected

Better Utilization of Protein Resources

Total Protein Available from Concentrate in India

is estimated to be around 8.5-9 million tones

Which can Support production of only 0.45 million

tonnes of Milk Proteins by the present mode of its

Utilization

Studies showed that when Protein Degradation in Rumen

is controlled and it is made to Bypass, the same can

support the production of 1.72 million of Milk Proteins

( Sharma, K. 2003)

Conclusion

Increases DMI Increase in milk production ( 10-15%) Increase in B. W. Gain EAA reach to SI Microbial Protein Synthesis; is Energy dependent Process Use of NPN should be Optimized

RDP/ UDP ratio should be Optimized