Concept of By-pass nutrients

Bypass Protein

Vishnu Vardhan Reddy.PTVM/2015-029

Department of Animal nutritionCollege of Veterinary Science, TirupatiSri Venkateswara Veterinary University

Definition of Bypass Protein• ‘‘Rumen protected’’ has been defined by the

Association of American Feed Control Officials (Noel,

2000) as:

‘‘A nutrient(s) fed in such a form that

provides an increase in the flow of that nutrient(s),

unchanged, to the abomasum, yet is available to the

animal in the intestine.’’

(NRC 2001 Pg.no:53)

Sources of Bypass Protein

1. Naturally Protected Proteins2. Heat Treatment3. Chemical Treatment4. Esophageal Groove5. Post Rumen Infusion (Fistula)6. Encapsulation of Proteins7. Amino Acids Analogs8. Lowering Ruminal Protease Activity9. Decreasing Retention Time in Rumen

Naturally Protected ProteinsFeed UDP %

Maize (grain) 65Barley 21( 11-27)Sorghum 52Bajra 68Oat grain 14–20Wheat grain 20–36Cotton seed meal 41–50Linseed meal 11–45Ground nut meal 30Rapeseed meal 23Soybean meal 28 ( 15–45)Sunflower meal 24Subabul 51 – 70

Feed UDP %Blood meal 76 – 82Fish meal 71 – 80Meat meal 53 – 76Brewers dried 53Corn gluten 53Wheat bread 29Corn silage 27Rice straw 63Wheat straw 45Para grass 52Cow pea 32 – 45Berseem 37 – 52Alfa-Alfa 28

(NRC, 1985; Dutta et. al., 1997)

Heat Treatment

• Heat processing of feed decreases protein

degradation in the rumen by denaturing proteins

and the formation of protein–carbohydrate cross-

links called as Maillard reactions and protein–

protein cross-links.

(Animal Nutrition by McDonald seventh edition Pg.no:566)

• According to the article “Estimates of protein fractions of various heat-

treated feeds in ruminant production” by Ho Thanh Tham, Ngo Van Man

and T R Preston.

Experiment feeds:

Cassava or Tapioca (Manihot esculenta, Crantz) (CLM)

Sesbania (Sesbania grandiflora) (SG)

Leucaena or Subabul (Leucaena leucocephala) (LL)

Gliricidia (Gliricidia sepium) (GS)

Water hyacinth (Eichornia crassipes) (WH).

Processing methods:

Heat treatment

60°C, 100°C, 140°C for 2 hours at each temperature.

Effect of heating on Fraction A (g/kg CP) in leaf samplesFeed 60°C 100°C 140°C Decrease in %

Cassava 114 111 71 37.7 %

Sesbania 465 452 411 2.8 %

Subabul 390 375 333 14.6 %

Gliricidia 186 170 78 58 %

Water hyacinth 142 133 115 19 %

Effect of heating on Fraction B1 (g/kg CP) in leaf samples

Feed 60°C 100°C 140°C Decrease in %

Cassava 42 29 3 92.8 %

Sesbania 19 10 7 63 %

Subabul 126 42 25 14.6 %

Gliricidia 138 134 57 80 %

Water hyacinth 155 67 62 60 %

Effect of heating on Fraction B2 (g/kg CP) in leaf samples

Effect of heating on Fraction B3 (g/kg CP) in leaf samples

Feed 60°C 100°C 140°C Decrease in %

Cassava 663 651 359 92.8 %

Sesbania 434 450 309 63 %

Subabul 421 497 545 14.6 %

Gliricidia 553 503 244 80 %

Water hyacinth 213 263 15 60 %

Feed 60°C 100°C 140°C Increase in %

Cassava 93 109 429 78.3 %

Sesbania 45 54 241 81.3 %

Subabul 93 96 100 7 %

Gliricidia 22 84 494 95.5 %

Water hyacinth 437 485 760 42.5 %

Effect of heating on Fraction C (g/kg CP) in leaf samples

Feed 60°C 100°C 140°C Increase in %

Cassava 88 100 138 36.2 %Sesbania 36 34 32 -11 % (decrease)Subabul 37 55 60 38.3 %Gliricidia 101 109 128 21 %Water hyacinth 54 53 48 -11 % (decrease)

• Heating the leaves to temperatures of 140°C for 2 hours

reduced the proportion of the protein in the A and B2 fractions

and increased the B3fraction.

Conclusions

• According to the article ”Optimization of roasting

conditions for soybean cake evaluated by in situ

protein degradability and N-fraction method” by Snjay

kumar , t.k.walli, rajani kumari.

Feed sample:

Soybean cake

Treatment method:

Roasting at 140, 150, 160, 170°C for 30 min.

Different nitrogen (% of total N) fractions in raw and roasted

soybean, roasted at different temperatures for 30 min

A+B1 fractions are positively related to ECPDB2 fraction negatively related to ECPD

Sample A+B1(PBSN)

B2 (PBIN-NDIN)

B3 (NDIN-ASIN) C (ADIN) ECPD

Raw (Without roasting) 35.22 63.30 0.405 1.08 58.5

140°C/30 min 32.28 66.03 0.780 0.81 50.2

150°C/30 min 29.67 67.90 2.008 0.482 48.3

160°C/30 min 25.88 70.89 2.14 1.09 46.0

170°C/30 min 22.88 71.12 3.36 2.64 45.1

ECPD=33.63424026+0.89824611(A+B1)-0.174329925(B2)-

0.2541073921(B3)+1.398296608(C)

ECPD=46.256225949+0.8253277191(A+B1)-0.320238478(B2)-

0.419856296(C)

ECPD=42.73418756+0.810303725(A+B1)-0.251985896(B2)

ECPD=22.50895891+0.917516113(A+B1)

Conclusion

Roasting at 160°C for 30 min was optimum time and

temperature fro soybean to make it good bypass

protein

Chemical Treatment

1) Formaldehyde treatment

2) Lignosulfonate treatment

3) Xylose Treatment

4) Tannic acid treatment

Formaldehyde treatment

• Treatment of high quality proteins result in the

formation of cross-links with amino group and

makes the protein less susceptible to microbial attack

(Czerkawski, 1986). These bonds are highly stable in

the near neutral pH of the rumen but are readily

hydrolyzed in the acidic pH of the lower digestive

tract.

Action of Formaldehyde as follows:

1) Formation of methylol groups on terminal amino groups of

protein chain and epsilon amino group or lysine

2) Condensation of these groups with primary amide of group of

asparagine and glutamine, and guanindyl group of arginine.

the condensation results in formation of intermolecular and

intramolecular methylene bridges. These bridges are broken

down in acidic medium of abomasum with liberation of

formaldehyde (Frankel-Convat and Oleott 1948)

• According to the article “Effect of processing on protection of highly

degradable protein sources in steers” by M.Yugandhar Kumar and A.Ravi.

Experiment feeds:

Babul seed cake, Coconut cake, Dried poultry waste, Guar meal, Mustard

cake, Rape seed meal, Tobacco seed cake.

Processing methods:

Heat treatment at 125°C for 3 hours.

Extrusion cooking at temperature 100-120°C screw speed 300-320rpm,

feeder rate 10-12 rpm, and products were cut into 1.5 cm and sundried and

ground.

Formaldehyde treatment with 3.5 gm. HCHO/100 gm. of CP.

Animals used: Four Ongole X Holstein crossbreed steers.

Effect of treatment on protein degradation kinetics of protein supplements

UT HT HCHOT EC UT HT HCHOT EC UT HT HCHOT EC

Babul seed cake 37 30 25 40 32.9 31 31.2 27.8 30.1 39.1 43.8 32.2

Coconut ckae 31 15 18 30 36.8 25 30.5 29.1 32.2 60.2 51.5 40.9

Dried proultry waste 75 60 90 74 19.5 31 22.5 20.7 5.5 9.5 7.5 6.3

Guar meal 75 68 41 58 24.2 31 52.5 39.8 0.8 1.1 6.5 2.2

Mustard cake 71 15 11 55 20.5 26 27.9 37.4 8.5 58.8 61.1 9.6

Rape eseed meal 58 9 6 35 35.4 14 20 60.4 6.6 77.2 74 7.8

Tobacco seed cake 20 10 8 14 40.8 36 20.2 25.4 39.2 54.3 71.8 60.6

Protein supplementFraction 'a' Fraction 'b' Fraction 'C'

HT HCHOT EC HT HCHOT EC HT HCHOT EC HT HCHOT EC

Babul seed cake 18.92 32.43 -8.11 -6.08 -5.17 -15.5 29.9 45.52 6.97 12.68 19 3.57

Coconut ckae 32.61 17.12 20.92 -32.61 -17.12 -20.92 86.96 59.93 27.01 39.69 28.04 5.42

Dried proultry waste 20 -20 1.33 56.41 15.38 6.15 72.73 36.36 14.54 8.86 4.56 0.55

Guar meal 9.33 45.33 22.67 27.69 116.94 64.46 37.5 712.5 175 11.9 41.04 3.93

Mustard cake 78.87 84.51 22.54 27.8 36.1 82.44 591.8 618.82 12.94 60.85 64.07 4.59

Rape eseed meal 84.48 89.69 39.66 -61.02 -43.5 70.62 1070 1021.21 18.18 77.92 75.41 9.29

Tobacco seed cake 50 60 30 -12.5 -50.49 -37.75 38.52 83.16 54.59 10.7 42.45 14.35

%reductionin EPDProtein supplement

Fraction 'a' Fraction 'b' Fraction 'c'

% decrease on UT % increase on UT % increase in UT

Effect of treatment on degradation kinetics of protein fractions and effective protein degradability

Heat treatment is better for reducing EDP of Rape seed

meal, and Coconut cake Formaldehyde treatment for

mustard cake, Tobacco seed cake, Heat or HCHO treatment

for Babul seed meal, Rape seed meal, and Mustard cake.

Dried poultry waste and guar meal were resistant to

different processing methods.

Extrusion cooking was least effective of the three

methods.

Conclusion:-

• According to article “Effect of Varying Levels of Formaldehyde and

Heat Treatment on in situ Ruminal Degradation of Different

Vegetable Protein Meals” by Faran hameed and Talat naseer pasha

Feed samples:

Maize gluten meal (60%), Rapeseed meal, Sunflower meal, Cottonseed

meal

Treatment method:

Formaldehyde treatment 0.50, 1.00 and 1.50% levels

Autoclaving for 0, 30, 45 and 60 minutes at 15 pound steam pressure

Animals used: Fistulated male buffalo calf

% of RUP values of feed samples as treated by

formaldehyde and autoclaving at 24 hours incubationFormaldehyde

TreatmentMaize Gluten

Meal, 60%Rapeseed

MealSunflower

MealCotton seed

Meal

Un treated 66.82 ± 1.61 19.62 ± 1.18 7.15 ± 0.58 20.6 ± 1.390.5% 93.68 ± 1.06 74.62 ± 0.56 78.67 ± 3.24 46.29 ± 2.331% 97.09 ± 3.10 84.75 ± 0.87 68.9 ± 3.61 50.59 ± 1.54

1.5% 89.55 ± 0.85 89.75 ± 1.35 79.25 ± 1.27 49.46 ± 0.68

Autoclaving timing

Maize Gluten Meal, 60%

Rapeseed Meal

Sunflower Meal

Cotton seed Meal

Un treated 66.82 ± 1.61 19.62 ± 1.18 7.15 ± 0.58 20.6 ± 1.3930 min 87.27 ± 0.82 47.33 ± 1.43 9.61 ± 0.51 40.92 ± 1.7845 min 84.38 ± 1.68 41.53 ± 0.60 13.49 ± 0.85 43.0 ± 3.2660 min 91.09 ± 1.31 50.79 ± 0.61 15.89 ± 1.36 64.05 ± 2.82

autoclaving

Conclusion• To achieve higher commercial rumen by-pass values

maize gluten meal (60%) and sunflower meal should

be treated with 0.5% formaldehyde whereas, for

cotton seed meal heat treatment through

autoclaving for 60 minutes gave better results.

• While comparing both treatments formaldehyde

treatment is practicable and economical.

Lignosulfonate treatment

• In general, the term "Lignosulfonate" is used to

describe any product derived from the spent sulfite

liquor that is generated during the sulfite digestion of

wood and containing a percentage of lignosulfonic

acid or its ash as well as hemicellulose and sugars.

(Windschitl and Stern, 1988)

• Because lignosulfonates can bind and precipitate

protein, it was hypothesized that protein meal

treated with lignosulfonates could be rendered less

degradable in the rumen.

• It was concluded that heat and the presence of

wood sugars in the lignosulfonate preparation were

necessary for a positive response.

(Windschitl and Stern, 1988)

• According to the article “Effect of lignosulphonate treatment of

groundnut and mustard cake on ruminal protein degradability in

cattle” by G Mondal, T K Walli and A K Patra

Feed sample: Groundnut cake, Mustard cake

Treatment method:

Calcium lignosulphonate treatment at the rate of 0, 5, 6 and 7 percent

on fresh basis (91.5% DM) with the addition of 10 percent water (weight

basis of fresh samples)

Then the treated samples were heated to 95°C for 2 h in a hot air oven

(Wright et al 2005).

Experimental animal used: Three mature Fistulated male crossbred

cattle

Feed Level of treatment CP RDP UDP Post ruminal

digestibility EPD, % Digestible UDP

GNC

0 % 43.2 32.4 10.8 88.3 74.9 9.55

5 % 43.1 24.1 18.9 89.32 56 16.9

6 % 43.1 24.9 18.2 83.5 57.7 15.2

7 % 43.4 24.0 19.4 80.6 55.4 15.6

MC

0 % 33.2 24.5 8.69 85.7 74.1 7.43

5 % 33.5 23.5 9.97 85.4 70.2 8.51

6 % 33.2 22.4 10.8 86.7 67.6 9.36

7 % 33.2 20.1 13.1 83.6 60.6 10.9

Effect of lignosulphonate treatment of GNC and M) on rumen

degradable (RDP), undegradable (UDP) protein and digestible

UDP, and post ruminal digestibility (percent) of UDP (percent of

DM basis)

Conclusions• The effective ruminal degradability of GNC protein decreased at 5

percent LSO3 treatment and post-ruminal digestibility of GNC

protein decreased beyond 5 percent LSO3 treatment.

• The effective degradability of protein of LSO3 treated MC

decreased at 7 percent treatment and post-ruminal protein

digestibility was not affected up to 7 percent level.

• Therefore, from this study it can be concluded that GNC and MC

may be treated with LSO3 at 5 and 7 percent levels, respectively,

to reduce the ruminal CP degradability without affecting the post

ruminal protein digestibility.

Xylose Treatment• Combination of heat and xylose enhances non-enzymic

browning (Maillard reactions) due to the increased

availability of sugar aldehydes that react with the protein.

Cleale et al. found that treatment of soybean meal with

xylose (3 mol xylose/mol lysine) was effective in reducing

degradation of soybean protein by rumen microorganisms.

(Animal Nutrition by McDonald seventh edition Pg.no:566)

• According to the article “The Effects of Xylose

Treatment on Rumen Degradability and Nutrient

Digestibility of Soybean and Cottonseed Meals” by

P. Sacakli and S. D. Tuncer

Feed sample: Soybean and cottonseed meals.

Treatment method: water +heat (100°C for 2 hours )

+0.5% or 1% xylose

Experimental animal used: Three ruminally cannulated

Merino rams

Rumen degradability characteristics and effective degradability

values of crude protein of untreated and treated cottonseed meal

Feed Treatment a % b % C % /(h) Pe %

SBM

SBM 12.26 86.70 0.0476 54.60

SBM + WH 12.02 86.50 0.0360 48.20

SBM + 0.5% X 5.90 86.08 0.0247 34.30

SBM +1% X 5.55 74.23 0.0223 28.40

CSM

CSM 11.58 81.07 0.0178 32.90

CSM + WH 11.50 77.41 0.0193 33.10

CSM + 0.5% X 11.58 76.57 0.0218 34.80

CSM + 1% X 13.94 86.06 0.0130 31.60

a: the rapidly soluble fraction b: the potentially degradable fraction c: the constant rate of disappearance of b Pe: the effective degradation

ConclusionSBM proteins can be effectively protected from

degradation in the rumen by xylose treatment through

Maillard reaction, without negatively affected in vivo

digestibility of protein, whereas xylose treatment

appeared to be less efficient on CSM proteins.

Tannin treatment

• The main effect of tannins on proteins is based on

their ability to form hydrogen bonds that are stable

between pH 3.5 and 8 (approximately). These

complexes stable at rumen pH dissociate when the

pH falls below 3.5 (such as in the abomasum, pH 2.5-

3) or is greater than 8 (for example in the

duodenum, pH 8).(S. J. Bunglavan and N. Dutta)

• According to the article “The Formation of ‘Ruminal Bypass

Protein’ (In Vitro) by Adding Tannins Isolated from Calliandra

calothyrsus Leaves or Formaldehyde” by Elizabeth wina,

Dindin abdurohman

Experiment material: Crude tannins were isolated from C.

calothyrsus leaves protein source is obtained form freeze dried

gliricidia leaves, milled soybean meal and casein.

Treatment method: Crude tannins at the level of 0, 10, 20, 30,

40 and 50 mg to each 0.5 gm. of protein source in test tubes.

Add 10 ml of rumen liquor to each tube and co2 gas id flushed

and incubated at 39°C for 48 hours.

Dry matter digestibility of casein, soybean meal and

gliricidia leaves at different levels of tannin isolate at 48

h of invitro incubation

Level of tannin isolate (mg/g

protein source)

Dry matter digestibility (%)

Casein Soybean meal G. sepium leaves

0 93.27 77.43 49.63

20 88.20 77.70 49.13

40 84.47 74.53 48.23

60 80.67 71.23 43.96

80 80.27 69.68 42.03

100 79.33 69.20 41.10

Instead of tannin 37% formaldehyde solution was added at the level of

2 g/100 g protein source in another test tube (BARRY, 1976).Compare

the results between Tannins(60mg/g) and Formaldehyde

Binding agent

Casein Soybean meal G. sepium leaves

DM CP DM CP DM CP

g/100 g substrate

Tannin isolate 6.7 5.3 24.3 27.9 24.4 34.4

Formaldehyde 77.2 81.4 50.6 54.1 23.5 32.1

Tannin isolated from C. calothyrsus can be used as a protein-binding

agent and has a similar activity with formaldehyde to bind forage

protein (Gliricidia sepium)

Conclusion

Esophageal Groove• This is normal function in young one. It is done/ good

for liquid proteins.

• Surgically fitted fistula after the rumen in the lower

tract of intestine is an easy method to avoid rumen

microbial degradation of proteins, so proteins/ amino

acids are available in the intestine.

Post Rumen Infusion (Fistula)

Encapsulation of Proteins

• Encapsulation of Proteins is usually done for good

Biological value proteins and for individual amino

acids. They can be given the form of capsule with a

combination of fats or fatty acids sometimes by

addition of carbonate, kaolin, lecithin, glucose etc.

Amino Acids Analogs

• Structural manipulation of amino acids to create

resistance to ruminal degradation is another

potential method for rumen bypass of amino acids.

• Analogs such as Methionine hydroxy, N-acetyl-DL-

Metionine, DLHomocysteine thiolactone-Hcl, DL-

Homocysteine, etc. have given satisfactory results.

Lowering Ruminal Protease Activity

• By depressing the proteolysis activity of the rumen

microbes we can slow down the protein degradation

within the rumen. Bacteria are the mainly

responsible for proteolytic degradation. So

antibiotics can be used to reduce the protein

degradation within the rumen.

Decreasing Retention Time in Rumen

• Less the time in rumen environment causes less

degradation. Faster pass of feed in the rumen is the

explanation. Factors influencing the rate of passage

include food intake, specific gravity, particle size,

Concentrate to roughage ratio, rate of rumen

degradation etc.

Importance of Bypass ProteinRequired for medium and high lactating and growing

animals mainly in early lactation.

Increase in Milk production by 10-15 %.

Good increase in live weight gain of meat purpose

animals.

Exposes essential and limiting amino acids directly to

Intestine.

Reduces Milk Production cost.

References1. Nutrient Requirements of Dairy Cattle Seventh Revised Edition, 2001.2. Animal Nutrition by McDonald seventh edition.3. Effect of processing on protection of highly degradable protein sources in

steers by M.Yugandhar Kumar and A.Ravi.4. Estimates of protein fractions of various heat-treated feeds in ruminant

production by Ho Thanh Tham, Ngo Van Man and T R Preston.5. Optimization of roasting conditions for soybean cake evaluated by in situ

protein degradability and N-fraction method by Snjay kumar , t.k.walli, rajani kumari.

6. Effect of Varying Levels of Formaldehyde and Heat Treatment on in situ Ruminal Degradation of Different Vegetable Protein Meals by Faran hameed and Talat naseer pasha

7. Effect of lignosulphonate treatment of groundnut and mustard cake on ruminal protein degradability in cattle by G Mondal, T K Walli and A K Patra

8. 8. The Effects of Xylose Treatment on Rumen Degradability and Nutrient Digestibility of Soybean and Cottonseed Meals” by P. Sacakli and S. D. Tuncer

7. Effect of lignosulphonate treatment of groundnut and mustard cake on ruminal protein degradability in cattle by G Mondal, T K Walli and A K Patra8. The Effects of Xylose Treatment on Rumen Degradability and Nutrient Digestibility of Soybean and Cottonseed Meals” by P. Sacakli and S. D. Tuncer9. The Formation of ‘Ruminal Bypass Protein’ (In Vitro) by Adding Tannins Isolated from Calliandra calothyrsus Leaves or Formaldehyde” by Elizabeth wina, Dindin abdurohman10. Role of bypass protein in ruminant production by Mayank Tandon, R.A. Siddique and Tanuj Ambwani11. Evaluation of Calcium Lignosulfonate-Treated Soybean Meal as a Source of Rumen Protected Protein for Dairy Cattle by p.m.windschitl and m.d.stern

THANK YOU

Vishnu Vardhan Reddy.PTVM/2015-029