Heat treatment and chemical treatments can increase the bypass protein content of feed ingredients fed to ruminants. Heat treatment through processes like autoclaving can increase the rumen undegraded protein fraction by denaturing proteins and forming protein-carbohydrate complexes. Chemical treatments using formaldehyde or lignosulfonate can also increase rumen undegraded protein by forming cross-links between amino acids or precipitating protein respectively, making it less susceptible to microbial breakdown in the rumen. The level of treatment and feed ingredient impacts the effectiveness at increasing bypass protein for ruminant digestion and nutrition.

1. Concept of By-pass
nutrients
Bypass Protein
Vishnu Vardhan Reddy.P
TVM/2015-029
Department of Animal nutrition
College of Veterinary Science, Tirupati
Sri Venkateswara Veterinary University

2. 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)

3. Sources of Bypass Protein
1. Naturally Protected Proteins
2. Heat Treatment
3. Chemical Treatment
4. Esophageal Groove
5. Post Rumen Infusion (Fistula)
6. Encapsulation of Proteins
7. Amino Acids Analogs
8. Lowering Ruminal Protease Activity
9. Decreasing Retention Time in Rumen

4. Naturally Protected Proteins
Feed UDP %
Maize (grain) 65
Barley 21( 11-27)
Sorghum 52
Bajra 68
Oat grain 14–20
Wheat grain 20–36
Cotton seed meal 41–50
Linseed meal 11–45
Ground nut meal 30
Rapeseed meal 23
Soybean meal 28 ( 15–45)
Sunflower meal 24
Subabul 51 – 70
Feed UDP %
Blood meal 76 – 82
Fish meal 71 – 80
Meat meal 53 – 76
Brewers dried 53
Corn gluten 53
Wheat bread 29
Corn silage 27
Rice straw 63
Wheat straw 45
Para grass 52
Cow pea 32 – 45
Berseem 37 – 52
Alfa-Alfa 28
(NRC, 1985; Dutta et. al., 1997)

5. 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)

6. • 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.

7. Effect of heating on Fraction A (g/kg CP) in leaf samples
Feed 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 %

8. 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 %

9. 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

10. • 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.

11. 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 ECPD
B2 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

12. 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

13. Chemical Treatment
1) Formaldehyde treatment
2) Lignosulfonate treatment
3) Xylose Treatment
4) Tannic acid treatment

14. 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.

15. 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)

16. • 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.

17. 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 supplement
Fraction 'a' Fraction 'b' Fraction 'C'

18. 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
%reduction
in EPD
Protein 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

19. 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:-

20. • 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

21. % of RUP values of feed samples as treated by
formaldehyde and autoclaving at 24 hours incubation
Formaldehyde
Treatment
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 ± 1.39
0.5% 93.68 ± 1.06 74.62 ± 0.56 78.67 ± 3.24 46.29 ± 2.33
1% 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 ± 1.39
30 min 87.27 ± 0.82 47.33 ± 1.43 9.61 ± 0.51 40.92 ± 1.78
45 min 84.38 ± 1.68 41.53 ± 0.60 13.49 ± 0.85 43.0 ± 3.26
60 min 91.09 ± 1.31 50.79 ± 0.61 15.89 ± 1.36 64.05 ± 2.82
autoclaving

22. 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.

23. 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)

24. • 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)

25. • 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

26. 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)

27. 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.

28. 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)

29. • 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

30. 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

31. Conclusion
SBM 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.

32. 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)

33. • 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.

34. 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

35. 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

36. 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)

37. 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.

38. 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.

39. 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.

40. 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.

41. Importance of Bypass Protein
Required 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.

42. References
1. 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

43. 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. The Effects of Xylose Treatment on Rumen Degradability and Nutrient
Digestibility of Soybean and Cottonseed Meals” by P. Sacakli and S. D. Tuncer
9. The Formation of ‘Ruminal Bypass Protein’ (In Vitro) by Adding Tannins
Isolated from Calliandra calothyrsus Leaves or Formaldehyde” by Elizabeth
wina, Dindin abdurohman
10. Role of bypass protein in ruminant production by Mayank Tandon, R.A.
Siddique and Tanuj Ambwani
11. 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

44. THANK YOU
Vishnu Vardhan Reddy.P
TVM/2015-029