Department 0f Animal Nutrition GADVASU, Ludhiana.

Course Title: Course No. Credit Hours

EVALUATION OF FEED STUFFS AND FEED TECHNOLOGY

ANN - 212 1+1

Animal feed technology The low grade roughages have poor nutritive value Nutritive value of crop residues Roughage Wheat straw Rice straw CP 2.8 3.7 DCP 0.0 0.2 CF 78.5 69.7 Silica 6.8 14.0

Poor quality roughages can be processed by different means to reduce the particle size of the lignocellulosic residue or other treatments thereby increasing the surface area available for enzymatic degradation.

Roughage processing

Physical method

Chemical methods

Biological methods

Wet methods 1. 2. 3. Soaking

Dry methods 1. Bailing 2. Grinding 4. Cubing 1 1. Alkali 2. Lime 3. Ammonia 4. Urine

Green chopping

Steam processing 3. Pelleting

5. Drying 6. Irradiation Physical methods

5. Urea

Green chopping- this is with green crops residues into 1-4 inches length by chaff cutter to reduce wastage, ease in handling and reduce selective eating by the animal. The FI and is not increased but digestibility increased slightly. Soaking- Sanni is done of wheat straw and concentrate mixture to increase intake. Steam processing- this is done to bagasse to increase VDMI and digestibility. There is physical disruption of cell wall structure resulting in hydrolysis of hemicellulose. There is loss of 25% DM and it is expensive. Bailing-or proper bailing the moisture should be 15-20%. They may be round or square shaped of 20-120 kg weight. Grinding-This reduces particle size and will encourage VDMI, daily gain due to higher DE available and dustiness. Grinding of hay reduced DMD due to 1. increased rate of passage, 2. reduced time for mastication. Hence there is decreased pH (due to fermentation) and less buffering by saliva, so acetate:propionate ratio decreases and the milk production drops in dairy cows due to low production of acetic acid in rumen. Pelleting-This reduces space requirement by 75%, increases VDMI, dustiness but involves extra cost. Pelleted roughage results in rapid rate of passage out of the rumen, less cellulose digestibility, less acetic acid production but higher ME utilization. Cubing is large pellets having diameter 2-3 inches and length 1-4 inches. Before cubing, grinding is not required but water is sprayed and then cubed. Drying- The moisture is kept at 14-15% drying under sun or artificial heat. The storage space required is reduced and it can be used during scarcity. Irradiation- Improvement can be by subjecting crop residues to high voltage Xrays (up to 12.5 105 rad.) which breaks cellulose and hemicellulose bonds to form oligosaccharides. Upon irradiation the ergosterol yields calciferol (vitamin D2). The process is not practical and expensive. 2

Alkali treatment-Use of alkali can reduce strength of intermolecular bonds which bind the cellulose fiber. Alkali also saponifies uronic acids and acetic acid esters. Silica is converted to soluble silicates. The method requires large amount of water as is impractical. The straw is soaked in 10 times its weight in 1.5% NaOH for 24 hours.The straw is then washed, dried and sued. It increases OM digestibility from 46-70%. Alternatives are 4kg NaOH per 160 kg wheat straw is sprayed, 15 min. treatment with hot 80-90oC NaOH solution. Ammonia treatment- This process increases the nitrogen content besides improving the degradability and VFA production. With 3% ammonia the CP of wheat straw increases from 2.7% to 8.8%, and cellulose digestibility increases from 48% to 60%. But ammonia is volatile so it has to be stacked and covered. Lime treatment- Calcium hydroxide solution 1.25% is sprayed and stacked for several days. The digestibility increases by 24-30%. Generally the DMI is reduced and excess calcium affects the utilization of other minerals especially P. Urine treatment-Animal urine can be source of urea to improve digestibility of fibrous material. It has the added advantage of adding calcium and phosphorus to the feed. Biological treatment- Since plant material have cell wall selected bacterial and fungal cultures can be used to improve the nutritive value of roughages. These microbes release appropriate enzymes, but microbes that can degrade lignin and not produce toxins need to be selected. The fast growth of microbes can increase the true protein content of the crop residues as well. The microbes used are Aspergillus niger and Coprinus fimetarius.

Every vegetable protein source has one or other anti-nutrients which can be destroyed during processing prior to use in the feed. Anti-nutrient substances are deleterious substances that arise during normal growth and metabolism of a plant. They can reduce the performance of the poultry and ruminants. The various anti-nutrients in feed ingredients are:
S. No.

1. 2. 3.

4.

5. 6. 7. 8. 9. 10. 11.

Anti-nutrient substance Occurrence Protease inhibitorsTrypsin Soybean seeds inhibitors Haemagglutinins (lectins) Legume seeds (castor bean, kidney bean, soybean) Glucosides a. Saponins Soybean seeds, Lucerne leaf meal Cassava b. Cyanogens (tapioca) root, linseed meal c. Glucosinolates Rape and mustard seed d. Estrogens Soybean seeds Phenols a. Gossypol Cottonseed meal b. Tannins Sorghum, rape and mustard meal, salseed meal, mango seed kernel, leucaena leaf meal, tamarind seed meal Phytate All vegetable feed ingredients Erucic acid Rape and mustard seed Argemone Rape and mustard seed meal Mimosine Subabul (leucaena) leaf meal Nimbidins Neem seed meal Oxalates Vegetable and animal feed sources Anti vitamins a. Anti vitamin A Lipoxygenase in soybean seeds b. Anti vitamin D Soybean seeds c. Anti vitamin E Kidney beans d. Anti vitaminK (Dicumarol) Sweet clover e. Anti vitamin B6 (Linatin) Linseed meal

Improving the nutritive value of protein sources S. No. Mechanism of action Deleterious effect 4

Detoxification

1.

2.

3.

a. Trypsin inhibitors form a complex with trypsin Pancreas produce more proteases (rich in methionine) Endogenous loss of methionine Haemagglutininsa. Binds with mannose (glycoproteins) of cells b. Binds to mannose of RBC and cause agglutination c. Attach to the intestinal cell lining resulting in poor absorption of nutrients Saponins- lower surface tension and form stable foam with proteins and cholesterol Haemolysis of RBC Alter cell permeability

Reduce growth Low production Pancreatic hypertrophy

Heat treatment Autoclave at 5lb/45 min Autoclave at 10lb/30 min Autoclave at 15lb/20 min Autoclave at 20lb/10 min Decrease in protein Cooking in water for 30 utilization min Poor growth Autoclaving for 30 min Soaking in water overnight Ammoniation Extraction with hot water or organic solvents (ethanol, methanol)

4.

Increases respiratory rate Inhibits the activity of certain enzymes (chymotrypsin) Lower growth Low egg production Cynogens- Intact glucosides not HCN inactivates toxic. On hydrolysis release cytochrome oxidase HCN. system

5.

Goiterogens- glucosinolates are hydrolyzed to 2 OH-3-butenyl isocyante and 5venyloxazolidinine-2-thionegoitroine, which suppresses iodine uptake by thyroid.

6.

Estrogens-Genistein isoflavone)

(an

7.

Gossypol- free gossypol at >150 mg/kg diet is toxic 5

Closed steam distillation with HCl at 100oC for 3 hrs. Cooking followed by discarding of cooked water Peeling of ski from tapioca tubers Goitre Extraction of Poor growth glucosinolates with hot Low egg production water, dilute alkali or Liver haemorrhages acetone Fatty liver Decomposition of glucosinolates with iron salts or soda ash Extraction of goitrogens with acetone or water or steam Poor growth Dry or moist heat Increased Zn in liver Solvent extraction and bone Increased deposition of ca, P and Mn in bones Poor growth Use iron salts at 4:1 ratio Low feed inake (iron and gossypol)

Binds with protein (lysine)

8.

Ascites Cardiac irregularity Reduce oxygen carrying capacity of the blood Olive green discoloration of yolk Tannins-bind with proteins Reduced growth Inhibit enzymes (trypsin, Poor egg production amylase, lipase, etc.) in intestines

Solid substrate fermentation with fungi (Aspergillus oryzae, Aspergillus janus)

9.

Phytate-six phosphorus molecules are bound to one phytic acid molecule. Ca, Mg salts of phytic acid is phytate. (Mn and Zn also bind to phytic acid) Phosphorus in feed is not available Phytase which hydrolyze phytate is not present in the intestine of birds.

Cold water processing Boiled water processing Treatment with dilute acid, alkali and salt Extraction with ether, acetone, ethanol and methanol Autocalving Increase requirement Microbial phytase of phosphorus and hydrolyze phytate and trace minerals release phosphorus, Ca and trace minerals

10. 11.

12.

13.

14.

Erucic acid is a polyenoic fatty Poor growth acid present in the oil of mustard Low feed intake Poor feed efficiency Argemone-it is through Swelling of leg adulteration with Argemona Diahhoea maxicana Atrophy of optic nerve edema Mimosine inhibits biosynthesis Redue growth of thryronine Weight loss Enlarged thyroid Poor reproductive efficiency Nimbidin Low feed intake Poor growth Low egg production Toxicity of internal organs Oxalates- soluble oxalates (Na Poor growth 6

Solvent extraction can remove oil and erucic acid Heating at 240oC for 15 min.

Urea treatment at 2.5% of need seed meal

16. 17. 18. 19. 20.

and K) and insoluble oxalates (Ca and Mg) Anti vitamin A-lipoxygenase oxidizes carotene Anti vitamin D

Decreased serum Ca Rickets Destruction of carotene Increase requirement of Vitamin D Anti vitamin E Increase requirement of Vitamin E Anti vitamin K Reduce utilization of vitamin K Anti vitamin B6- Linatine (a Reduces utilizationof peptide containing 1-amino-D- pyridoxine prolone in combination with glutamic acid)

Heat treatment Heat treatment Autoclaving

Extraction of linseed meal with water Autoclaving

Heat treatment is used effectively in the industry to destroy protease inhibitors, haemagglutinins, estrogens, anti vitamin A, D and saponins. Sun drying is used to reduce cynogens in cassava roots. Phytase and poly enzymes are used for phytates and NSPs.

Classification of feeds and fodders Advantages of classification of feeds 1. Group helps in the selection of cheaper feeds for formulating the diets. 2. For substitution of one/two feedstuffs from the same group. 3. Feeds of similar nutritive characters are grouped. International feed classes system. 1. Dry forages and roughages 2. Succulent forages ad pastures 3. Silage 4. Energy feeds 5. Protein supplements 6. Mineral supplement 7. Vitamin supplement 8. Additives 1. General classification- on the basis of the bulkiness and chemical composition. On the basis of crude fiber, feeds are classified as A. B. 1. 2. 3. 4. a. b. Roughages contain more than 18% CF and , 60% TDN. Concentrates On the basis of moisture content- Succulent which contain more than 80% moisture or Non succulent On CP content/type of plants Leguminous or Non-leguminous Green forages or Dry forages On the basis of nutritive values (on DM basis) Maintenance type contains 3-5 % DCP eg. Non-legume cereal fodders and their hay Production type more than 5% DCP eg. Leguminous and their hay 8

The roughages can further be classified on the basis of

c. Concentrates: a.

Non-maintenance type contains less than 3% DCP eg. straws and stovers. Energy rich feeds: These feeds contain more than 50% of starch in them. The crude protein is less than or equal to 20%. eg. brans, chunni, husks.

b.

Protenecious feeds: Contain more than or equal to 20% CP.

International feed vocabulary Origin: The origin of the parent plant material which may be specific or nonspecific. Part: The actual part of the parent feed material ie. roots, stems, leaves, seeds or their fractions given to the animals. Processes and treatments: The operations done on the part of food/feed material before feeding to the animals. Stage of maturity Cutting: Grades: International feed classes: Code 1 2 3 4 5 6 7 8 Class description on DM basis Dry forages and roughages Pastures, range plants and forages fed green Silages Energy supplements Protein supplements Mineral supplements Vitamin supplements Additives

International Feed Name is a complete description Class-IFNRice straw Rice Hulls Para grass Maize silage Groundnut cake 1-03-925 1-08-925 2-03-525 3-02-818 5-03-648

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Classification of feeds and fodders There are many different kinds of products that are used for feeding purposes. They may be grouped based on certain similarities in composition and feeding value.
Composition characteristics of different feed groups Legume(>10.5% protein, > 0.9% Ca) Mineral products (over 80% ash) Air dry feeds Over 80% DM Hays (18-34% fiber, 40-60% TDN, considerable carotene) Non-legume (6-10, 5% protein, < 0.9% Ca) Air dry roughage (> 18% fiber, < 60% TDN) Low grade roughage (28% fiber, most over 34%, <52% TDN, several < 40%, very little carotene, most under 6% protein) Air dry concentrates (< 18% fiber, > 60% TDN) Energy feeds (<18% protein) Protein feeds (>18% protein) Animal origin (> 47% CP, > 1.0% Ca, > 1.5% P, < 2.5% fiber)

Plant origin (< 47% CP, <1.0% Ca, < 1.5% P, > 2.5% fiber) Defatted (<17% EE) Oil seed (> 17% EE)

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Classification on the basis of moisture content

High moisture grain (70-80% DM, > 7% protein, some fiber) Molasses (60-80% DM, < 7% protein, 0.0% fiber) Haylage (45-60% DM, similar to hay in composition) Silage ( 25-45% DM, medium carotene) High moisture feeds (< 80% DM) Fresh forages (15-30% DM, high in carotene) Wet by-products (10-25% DM, > 3% fiber, little carotene) Root crops (9-30% DM, < 3% fiber) Fresh whole or skimmed milk (9-13% DM, 0.0% fiber)

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Definitions of feeds and fodders Food: An edible material Ration: Amount of food/feed allowed or given to an animal for a period of 24 hr. Forages and Roughages: The term roughages are used to designate the feeds that are high in fiber and low in net energy. Hay: Hay is the product obtained by drying the tender stemmed leafy plant material or forage crop in sun or shade Straw: The by-product of any cereal, millet or legume crop left over after harvesting, threshing and removal of grains or pulses Fodder: The aerial parts with ears, husk or heads are called fodder. Stover: The aerial part without ears, without husks or heads is called stovers Bagasse: Hull: Fibrous material left over in sugar factories after the extraction of juice from the sugarcane. The outer covering of beans, peas, cottonseed Husk: Dry outer covering of grains and gram eg. rice husk, gram husk Shells: The hard outer covering of nuts eg. ground nutshells Hulls, husks and shells are rich in crude fiber, low in CP and are light and bulky. Corn cobs: These are left after the removal of grains from the corn.

Pasture grasses : eg. Cenchrus ciliaris, C. Setigerus, Heteropogon Contortus Pasture legumes: eg. Stylosanthes hamata, S. Scabra, Siratro, Butterfly pea Soilage: Pastures when cut down and fed as green to an animal in its own stall Soiling or Zero grazing: The method of feeding an animal with soilage is called soiling or zero grazing.

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Chemical composition of common feedstuffs (in %)

Feedstuff DM CP Cereal straws Rice straw 90 3 Wheat straw 90 3 Oat straw 90 4 Ragi straw 90 3 Jowar straw 90 5 Legume straws Gram straw 90 6 Groundnut straw/ 90 8 Haulms Hays Indigenous grass 88 5 Improved grass 88 7 Legume (cowpea, 88 15 Lucerne, berseem) Green fodders (on wet basis) Non-leguminous Maize 25 2 Oat 25 3 Jowar 25 1.5 Napier bajra hybrid 25 2 Guinea grass 25 1.5 Dub grass 35 3.5 Sugarcane tops 35 2.5 Leguminous 4.5 Cowpea 20 4.5 Lucerne 20 4.5 Berseem 15 3 Cereal grains Maize 90 11 Oat 90 9 Legume seed Cowpea 90 25 Brans Deoiled rice bran 90 12 Wheat bran 90 13 Oil cakes Til cake/ Sesame 90 45 cake Groundnut cake 90 52 Coconut cake 90 24 Cottonseed cake 90 23 Mustard cake 90 36 Linseed cake 90 31 EE 1 1 1 1 1 0.5 1 1 1.5 2 CF 32 38 36 36 34 45 41 35 30 30 NFE Ash Ca 49 46 52 52 50 36 42 47 51.5 41 15 12 6 8 10 13 8 12 10 12 0.5-0.6 P 0.12-0.14

1.0-1.4

0.08-0.15

0.3 0.35 1.5 0.07-0.09

0.2 0.25 0.25 0.04-0.05

0.6 0.8 1 0.5 0.5 1 1 1 1 0.5 2.5 1.5 2 2 2.5 10 8 10 9 11 6

8 6 9 10 8 8 8 5 5 4 2 5 5 13 13 5 7 13 24 10 10

13 14 12 11 14 17.5 21 5.5 5.5 8.5 82.5 80.0 64 68 64.5 29 27.5 45 37 33 43

1.4 1.2 1.5 1.5 1 5 2.5 0.3-0.5 4 4 4 2 4.5 4 5 7 11 5.5 8 7 10 10 0.04 0.16 0.08 0.1-0.3 0.5 0.35 0.41 1.5 0.8-1.2 1.0 0.05-0.10

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Nutritive value of common roughages (average values) (on as fed basis) in %

Feedstuff Dry roughages Cereal straws (Rice, Wheat) Legume straws Groundnut straw Cereal hays Jowar hay Legume hay Green roughages Cereal fodders (Maize, Jowar, Bajra) Green grasses (Hybrid napier, Para, Guinea) Sugarcane tops Legume fodders (Berseem, Lucerne, Cowpea) DM 90 90 88 88 20 25 25 15 DCP 0 8 3 10 1 1.5 0.7 3 TDN 44 53 51 60 13 12 10 12 0.07-0.09 0.3-0.5 0.04-0.05 0.05-0.10 Ca 0.5-0.6 1.0-1.4 0.3 1.5 P 0.12-0.14 0.08-0.15 0.2 0.25

Nutritive value of common concentrate feedstuffs (on as fed basis) in %

Feedstuff Maize Jowar Wheat Deoiled ric bran Wheat bran Gran husk Chuni (mixture of broken gram and husk) Tapioca waste Molasses Groundnut cake (exp) Groundnut cake (extr) Til/Sesame cake DM 9 9 13.2 12 14 6 4 4.8 45 52 47.8 33 39 39 24 36 45.8 50 50 DCP 7.4 8 10.2 6 10 0.5 6-8 2 1 42 46 38 23 32.8 18 19 27 42 45 45 TDN 84.9 75 79.2 66 68 55 60 64 54 71 70 78 71 72.5 70 81 74 78 65 70 0.6 8-10 29 1 4-5 12.6 Ca 0.04 0.08 0.1-0.3 P 0.35 1.5 0.8-1.2

0.2

0.7

Sunflower cake (ext)

Cottonseed cake (extr, decorticated and delinted) Cottonseed cake (undecorticated) Coconut cake (extr) Mustard/rapeseed cake (extr) Soybean meal (extr) Fish meal Meat meal Bone meal

15

Calcite

Dicalcium phosphate

36 21

18.5

BIS specifications for certain concentrate ingredients (on % DMB)

Feedstuff Moisture (Max) CP (Min) EE (Min) CF (Max) AIA (Max)

Groundnut cake (GNC) GNC (exp) grade 1 grade 2 GNC (ext) grade 1 grade 2 Cottonseed cake (CSC) Decorticated CSC Decorticated CSC grade 1 Decorticated CSC grade 2 Undecorticated CSC Undecorticated CSC grade 1 Undecorticated CSC grade 2 Rapeseed cake type 1 Rapeseed cake type 2 Coconut cake grade 1 Coconut cake grade 2 Linseed cake (LC) LC expeller grade HF LC expeller grade LF LC (extr) sp. grade LC (extr) grade A Maize gluten grade 1 Maize gluten grade 2 Wheat bran Rice Polish Calf starter meal Calf growth meal

8 8 8 8 8 8 8 8 10 10 11 11 8 8 10 10 10 10 12.5 10 10 10

48 43 51 47 40 35 24 20 37 35 22 18 29 31 33 29 45 23 13 11 23-26 22-25

7 6 7 6 7 5 5 8 6.5 11 8 5 1 1.5 4 3 15 4 4

8 12 7 10 12 15 22 26 8 8 12 10 10 10 9 11 3 8 12 4 7 10

2.0 2.5 2.5 2.5 2.0 2.5 2.0 2.5 1.5 2.0 1.5 1.5 1.5 1.2 2.5 2.5 0.5 0.5 0.25 1.5 2.5 3.5

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Food Energy Measurements Evaluation of feeds for energy The potential of any feed for the supply of a nutrient can be done by the chemical analysis. The actual value can be arrived at by providing the allowances for inevitable losses occurring due to 1. 2. 3. Digestion Absorption Metabolism

The major nutrients of the feed in the body are required for the synthesis of body tissues, synthesis of milk, wool, eggs and for work production. All these functions involve the transfer of energy from chemical energy to heat energy or conversion of chemical energy to another form eg. synthesis of fat from dietary carbohydrates. Therefore the ability of a feed to supply energy is of great importance in determining the nutritive value. Bioenergetics: The topic of energy and its metabolism is called bioenergetics. Energy: The ability to perform work is called energy. It is an abstraction that can be measured only in reference to standard conditions. Energy is derived from most organic nutrients/compounds of the feed. The animals derives energy by partial or complete oxidation of molecules ingested and absorbed from the diet or from the metabolism of body reserves ie. fat, carbohydrates and proteins. Energy transfer from one chemical reaction to another occurs primarily by means of high energy bonds found in such energy compounds as ATP and other related compounds. Work: It is the product of a given force acting through a given distance. For nutritional usage chemical energy is typically measured in terms of heat produced upon oxidation and is expressed as calorie. Calorie: It is the amount of heat required to raise the temperature of 1 gram of water by 1oC from 14.5 to 15.5oC. 1 calorie = 4.184 joules.

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Kilo calorie: Heat required to raise the temperature of 1 kg water by 1oC. 1 Kcal = 1000 calories. 1 Mega calories = 106calories = 1000 Kcal. BTU (British Thermal Units): The amount of heat required to raise the temperature of 1 lb. of water by 1oF 1 Kcal = 4 BTU In UK Kilo joules has been adopted. International system of units (SI) has been adopted since 1970. SI unit of energy is Joules (J) 1 Kcal = 4.184 K joules 1 KJ = 0.239 Kcal. The quantitative measure of chemical energy of foods is measured by converting it into heat energy and determining the heat produced. The first measurement in nutritional evaluation of feeds is Gross energy. Gross Energy (GE) : It is the amount of heat given off when a given feed, food, milk, meat, egg or other substances are completely burnt/oxidized (to its ultimate oxidation products ie. CO2, water and other gases). GE is equivalent to heat of combustion. It is measured by burning a known quantity of material in a bomb a calorimeter in the presence of sufficient amount of oxygen. It is expressed as per unit basis of the material. GE of different materials varies depending upon the composition of material ie. protein, fat or carbohydrates. The chemical energy varies inversely with C/H ratio and CO2 and N contents. Gross energy of some tissues, feed Food/Material GE Kcal/g Carbohydrates Av. (4.15) Glucose 3.74 Starch 4.18 Glycerol 4.31 Proteins Av. (5.65) Casein 5.90

Food Sucrose Cellulose Beef muscle

GE Kcal/g 3.94 4.18 5.30

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Gluten Glycine Urea

6.0 3.00 2.52

Egg albumin Alanine Tyrosine Uric acid

5.70 4.35 5.91 7.74 9.10 9.40 3.49 5.95 9.53 13.3 4.50 4.50 5.10 or 1 atom of oxygen per atom of

Fats Av. 9.4 Beef fat 9.40 Butter fat Coconut oil 8.90 Corn oil Propionic acid 4.96 Acetic acid Palmitic acid 9.35 Butyric acid Oleic acid 9.50 Stearic acid Ethyl alcohol 7.11 Methane Feeds Maize 4.40 Wheat bran Grass hay 4.50 Oat straw Soybean emal 5.50 Linseed meal Example: Glucose has empirical formula of C6H12O6 carbon

Fat molecule has formula C57H104O6 or oxygen atoms to carbon atom has ratio of 1:9.5. Thus fat require more oxygen for oxidation and produces more heat. The gross energy values by themselves are of little practical value for describing the energy value of feeds for use by the animal because these do not consider the ability of the animals to utilize the feed. Increase in protein and/or fat increase the GE. While increase in ash or mineral contents decrease the energy value. Gross energy feed intake (GEi)- It is the gross energy of the feed consumed GEi = FI (DM basis) X GE of feed per unit dry weight GE losses from the animal body- A portion of the feed is not digested and excreted in the feaces. A portion of it is also lost in the form of combustible gases (methane chiefly) formed during fermentative digestion of nutrients. A portion is lost in urine as urea, uric acid and creatinine due to incomplete oxidation of proteins. A portion of it is lost as heat due to digestion and metabolism of nutrients. Feacal energy (FE)- It is the gross energy of feaces. It consists of gross energy of undigested feed, metabolic fractions (digestive fluids and abraded mucosa) present in the feaces.

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Gaseous products of digestion (GPD): It includes the combustible gases produced in the digestive tract by the fermentation of the ration eg. H, CO2, CO, H2S, acetone, ethane and methane. Methane makes up the largest portion of these gases as is significant in ruminants. It is higher when animals are fed roughages than concentrates. The values are lower as the level of feed intake increases. Generally the gaseous losses are 7% of GE. In the non-ruminants it is negligible. A substantial amount of cellulose is fermented in the caecum of horses. The gas production in horses is less than in ruminants. As the determination of gases is expensive and laborious it is calculated as Sheep E= 2.41 X + 9.80 (Swift et al. 1948) Cattle E = 4.012 X + 17.68 (Pennsylvania workers) Where E is methane in gms and X is digested carbohydrates in 100 gm. Methane contains 13.34 Kcal per gm. Urinary energy (UE) : UE is the GE of the urine. It included the non oxidized portion of the absorbed nitrogenous products, primarily urea in mammals and uric acid in birds and the energy present in the endogenous fraction of the urine (UEe). It is 2-3% of GEi in pigs and 4-5% of GEi in cattle. Under true energy distribution system, FEm and UEe are part of the maintenance requirement of the animal. Metabolizable energy (ME): It is the portion of the total energy ingested capable of being utilized by the body. Ruminants : ME= GEi FE GDP UE Non-ruminants : ME = GEi FE UE Nitrogen corrected ME (MEn) : The ME value of the feed also depends if the amino acids supplied by the protein is retained or the amino acid is deaminated and the N is excreted as urea/uric acid in the urine. Thus, the ME values are corrected to zero N balance (NB) by deducting for each 1 g of N retained or by adding for each 1 g of N catabolized. The value per gm is Pigs = 6.7 Kcal (28KJ) Poultry = 8.1 Kcal ( 34KJ) Ruminants = 7.4 Kcal ( 31KJ)

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Ruminants MEn = GEi FE GPD UE (NB X 7.4) Total digestible nutrients (TDN) : It gives the apparent digestibility of feed and is expressed by weight or as percent. Digestibility coefficients are used to calculate TDN. Digestible EE is X by 2.25 as fats have 2.25 times more energy than carbohydrates. TDN = DCP + DCF + DNFE + (DEE 2.25) Proximate % Dig. Coe. Digestible TDN

principles nutrients CP 20.11 75.0 15.08 CF 16.25 73.9 12.01 NFE 40.99 80.6 33.03 EE 3.34 53.9 1.8 2.25 = 4.04 64.16% Net energy (NE): It is the energy available for useful purpose. NE = ME- HI NE is used for maintenance (NEm) or maintenance (NEm) and production. It is the energy required to keep the animal in equilibrium. In winters some of the HI is also used for NEm. The NEm of a producing animal is higher than a non producing animal due to hormonal variation and difference in voluntary activity. NEm includes the energy required for basal metabolism, activity and to maintain body temperature. Net energy for production (NEp) or NE gain : NEp is the fraction of energy required for growth, fattening, milk, wool, egg, etc. GE of feed Feed origin FE Metabolic origin DE Gaseous products of digestion Feed origin Urinary energy Endogenous origin

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ME HF HI HNM

NE NEp or NEg NEm Scheme of energy utilization Basal metabolism is the minimum energy required by a non producing animal at rest to carry out the essential life processes like breathing, circulation of blood, maintenance, repair of tissues etc. Energy for voluntary activity (VAE) : Energy required for getting up, standing, moving, etc. HBW is the extra energy need to keep the body warm when the temperature is cold, and HBC is the extra energy needed to keep the body cool when the environmental temperature is hot. Total heat produced (HP) = HI + NEm NEg/NEp of energy retention = ME HP Physiological fuel value (PFVs): These are calorific value given by Atwater for humans to calculate GE that is available to the body. It is as important as ME. It is derived as ME X digestibility coefficients. In protein 1.25 Kcal is subtracted from GE of protein as allowance for loss of energy through urine. Nutrient Carbohydrate Fat GE Kcal/g 4.15 9.40 Digestibility coefficient 98 95 PFV Kcal/g 4.0 9.0

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Protein

5.65 1.25 = 4.4

92

4.0

These values are not appropriate for ruminants as they have less digestibility coefficients and the factor 1.25 is for humans eating mixed diet. In ruminants hippuric acid is excreted in the urine. Heat increment (HI) : It is the increase in heat production after consumption of feed when the animal is in a thermoneutral environment. HI is due to heat of fermentation (HF) and heat of nutrient metabolism (HNM). HF is the heat produced in the digestive tract as a result of microbial action during intermediary metabolism of absorbed nutrients. It corresponds to the specific dynamic action or calorigenic effect or thermogenic action. Rubner coined the word SDA. HI is generally wasted except in winters. Nutritive ratio (NR): It is the ratio of digestible protein to the sum of digestible carbohydrate and fat where fat is by 2.25. It is also called albuminoid ratio. It is important because the proteins can perform functions that cannot be done by other non protein nutrients. Feeds rich in protein have a narrow nutritive ratio and feed low in protein are wide in nutritive ratio. A ratio of 1:9 is suitable for horses and cattle, 1:6 for early fattening, lactation, working animals and 1:0.7 for young stocks. eg. DCP= 42, DEE= 6, DCF= 1 and DNFE= 14.5 so NR= (6 2.25) + 14.5 + 1 ----------------------= 0.70 and NR is 1:0.70 42 Nutritive value index (NVI): The feeding value of roughages depends on its digestible nutrients as well as its voluntary intake. Crampton and his coworkers coined the term NVI in 1960. NVI is calculated by 42 multiplying the intake relative to some defined standard forages which is taken as 100 and TDN value of the roughages. The consumption of standard forage is 80 g/kg metabolic body size. eg. a ram of BW 50 kg consumes 1.1 kg DM so intake/W 0.75 = 1.1/500.75 = 1.1/18 = 58.5 g so relative intake of roughage is 58.5 100/80 = 73%. The TDN of the roughage is 60 % so NVI = 73 60/100 = 43.8%

23

True metabolizable energy value of feeds for poultry: The measurements of energy value of diet is given by DE, ME and NE. In birds ME is calculated as excreta (feaces and urine) is single material. It is measured by feeding control grp. 45.7 % glucose and test grp 5.7% glucose + 40% test material. Both the groups are given 54.3% premix of other nutrients. The diet is fed for 15d followed by 4 d metabolic trial. Representative samples are collected of total feaces or by using chromic oxide marker method. Factors for interconversion of TDN to DE; DE to ME Schneider gave the formula 1 kg TDN = 4.38 Mcal DE Crampton and associates and Swift suggested 1 kg TDN = 4.41 Mcal DE . The variation is due to species, type of ration. ME is calculated as 0.82 DE in ruminants. The ratio of DE to ME varies as 0.82 for roughages, 0.85 for cereals and 0.79 for oilseed cakes and meals. NRC uses ME = 0.82 DE

eg. Calculation of DE Proximates % CP EE CF NFE Ash Water Total /g 10 3 10 65 2 10 100

Av. calorific value/g 5.65 9.40 4.15 4.15

GE Kcal 56.5 28.2 41.5 269.8 396 3.96

Dig. Coe DE, Kcal % 75 90 50 90 42.5 25.4 20.8 242.8 331.5 3.32

eg. Calculation of ME Feed DMI kg/h/d Soybean 0.795 hay Soybean 0.674

FI

4.333 4.345

Energy/kg DM, Mcal ME Mcal Losses Feaces Urine Methane 2.033 0.196 0.208 1.896 2.676 0.042 0.229 1.398 24

straw eg. Actual energy balance Partitioning of energy in the animal system DMI = 1,829 kg GEI = 35.0 MJ (1 MJ= 0.239 Mcal) FE = 13.5 MJ UE = 1.2 MJ GE = 2.4 MJ HI = 7.0 MJ DE ME NE = = = 35 13.5/1.829 = 11.8 MJ/kg 35 (13.5 + 1.2 + 2.4)/1.829 = 9.8 MJ/kg 35 - (13.5 + 1.2 + 2.4 + 7)/1.829 = 6.0 MJ/kg

Starch Equivalents or Starch Values O. Kellner introduced the concept of starch equivalent which is same as net energy value of feeds. SE is expressed in kg of starch which is the source of energy to the animals. Kellner added pure carbohydrates, fat and proteins to a maintenance diet and calculated the amount of pure nutrients required to produce unit of pure body fat. He expressed this fat producing power of feed as the kg of starch required to produce the same amount of fat as 100 kg of feed. Direct and Indirect Calorimetry Heat production can be measured by animal calorimeter which is known as direct calorimetry. Heat production can be made by estimating the respiratory exchange of gases using a respiratory chamber. Direct calorimetry: It is measured directly in a respiratory calorimeter or animal calorimeter. Lavoisier was first to recognize that heat is produced by oxidation of food. Lavoisier and Laplace (1780) devised the animal calorimeter. The animal was enclosed in a chamber surrounded by ice and the amount of ice melted in a given period of time was recorded as a measure of the amount of heat given off by the animal. In Adiabatic type the ice is replaced by water. The latest type is the gradient layer calorimeter.

25

Respiratory calorimeter combines the features of a respiration chamber and a calorimeter. It was first devised by Rubner in 1881 for dogs. In the respiration calorimeter it is possible to account for the intake of feed, water and oxygen and the outgo of solid, liquid and gaseous excreta and of the heat eliminated. It is presumed that the heat lost from the body is equal to the heat produced. The heat is lost from the body is as radiation, conduction and convection (sensible heat loss) and by evaporation of water from the skin and lungs (evaporative heat loss).The animal calorimeter/respiration calorimeter is an air tight chamber and has a special insulated construction which prevents the chamber from gaining or losing heat. The heat loss of the animal kept in the chamber is measured by the rise in temperature of the cold water flowing in various pipes. In addition to the metabolic trial the CO2 and methane loss are accounted for. The HI is estimated as the difference in the heat produced at two levels of intake. Two levels are taken to account for the basal metabolism which remains the same. The extra heat produced at higher level of intake is due to the higher feed intake. The procedure for estimating energy by this method is very expensive. Therefore indirect calorimeter is used. That is respiration calorimeter is replaced by respiration chamber. Example: Data of Direct calorimetry Vol. of water circulating through coil = 2000L Av. inlet temperature=18.5oC Av outlet temperature= 19.1oC Increase in wt. of absorbent= 1256g Calculations: Heat removed through water = 2000 0.6= 1200 Kcal Heat removed through evaporation (585 cal. Per g of water evaporated)= 585 1256= 735 Kcal Total= 1200 + 735 = 1935 Kcal. Indirect calorimetry by the measurement of respiratory exchange (Respiratory quotient) The main form in which the energy is stored is as fat and proteins as glycogen stores are negligible. 1gm molecule of oxygen occupies 22.4 L at NTP. So energy obtained

26

solely from glucose is 675 4.2 6 22.4 = 20.98 KJ (5.014Kcal) of heat. This value is known as thermal equivalent of oxygen. Respiratory quotient: This is the ratio of volume of CO2 produced to volume of O2 used. So RQ of carbohydrate is 1 (6O2 and 6 CO2) and fat is 18 CO2/26 O2= 0.692 and for protein is 0.8. The nitrogen is excreted as urea and small amount of other incompletely oxidized compounds such as hippuric acid, creatinine, allantoin, ammonium salts, uric acid etc. are formed so RQ is 0.83. If the RQ of an animal is known, the proportion of fat and carbohydrates oxidized can be determined from the standard tables. Example: of indirect calorimeter Oxygen consumed CO2 produced Urinary N excreted Heat from protein metabolism Protein oxidized 14.8 6.25 Heat produced 92.5 4.3 Kcal Oxygen used 92.5 0.96 CO2 produced 92.5 0.77 Heat from carbohydrate and fat metabolism Oxygen used 392.2-88.8 CO2 produced 310.7-71.2 Non-protein RQ Thermal equivalent of O2 at RQ 0.79 (from table) Heat produced 303.2 4.789 Total heat produced 398.8 1452 In the combustion of 1 g of protein 0.96 L of produced. The value of RQ gives the proportion of nutrient burned eg. if 1 then carbohydrates, if close to 0.7 then more fat and close to 0.8 then more protein. If RQ is 1.4 then the carbohydrate is being converted to fat and RQ below 0.7 indicates fasting or hibernating animals. 392 L 310.7 L 14.8 g 92.5 g 398.8 Kcal 88.8 L 71.2 L 303.2L 239.5 L 0.79 4.789 Kcal/L 1452 Kcal 1850.8 Kcal O2 is used and 0.77 L of CO2 is

Apparatus for Measuring Respiratory Exchange

27

An animal respiration chamber is commonly used. The exchange of gases is done by a face piece that collects the air for analysis. The chamber can measure the heat lost in perspiration, gaseous exchange though intestine and pulmonary losses. They are of two types. The close circuit designed by Regnault and Reiset and the open air developed by Pettenkofer. In the close circuit large amount of absorbent (soda lime) is used. Eg. Indirect calorimetry Data: decrease in wt. of O2 cylinder= 3.293 g Increase in wt of CO2 absorbent= 3.703 g Vol of CO2 =3.703 0.5094 = 1.886 L Vol. of O2 = 3.293 0.6998 = 2.304 L RQ = 1.886/2.304 = 0.819 Calorific value of 1 L O2 at RQ 0.819 = 4.824 Kcal Calories/10 min. = 4.824 2.304 = 11.11 Kcal Total heat produced per hr. = 66.66 Kcal (1g CO2 = 0.5094 L; 1 g O2 = 0.6998 L) In open air the air enters at a metered rate and the air is sampled at entry and exit point. Thus CO2 produced, O2 consumed is estimated. The heat production is estimated and energy retention is calculated as the difference between ME intake and heat production. Measurement of energy retention by the carbon and nitrogen balance technique. The main form in which the energy is stored is as fat and proteins. The quantity of fat and protein stored is calculated by C and N balance. The energy retained is calculated by multiplying the nutrient with its calorific value. Determination are made of the N and C in the feed, feaces and urine and of the C I the gaseous output. Example : carbon and nitrogen balance Amount Feed hay 6.988 kg Nitrogen (g) Intake Outgo 56.4 Carbon (g) Intake Outgo 2831.7 28

linseed meal Excreta: faeces

0.400 kg 16.619 kg 4.357 kg 37 g 4.73 kg 142 g

21.9 -

33.5

172.6 -

1428.7

urine 32.4 124.2 brushings 1.3 8.0 CO2 1290.2 CH4 106.6 Total 78.3 67.2 3004.3 2957.7 Gain Carbon=46.6 g nitrogen=11.1 g Protein has 16.65% nitrogen so protein is 11.1 100/16.65= 66.6 g Protein contains 52.54% carbon so this much protein has 66.6 52.54/100=35 g carbon. The total gain in carbon is 46.6 g so carbon in fat is 11.6 g. Since fat has 76.5% carbon so, the fat is 11.6 100/76.5=15.2 g. So energy retention is (66.6 5.64) + (15.2 9.39)= 518 Kcal. If the ME intake is known then ME intake- energy retention produced. The advantage of the C and N balance technique is measurement of oxygen consumption or RQ is not required. In the slaughter method the gain in protein and fat is measured by estimating the GE before and after the experiment. Thus HP can be measured by animal calorimetry, indirect calorimetry by respiratory chamber, carbon nitrogen balance, by slaughter technique and by Brouwer equation.

29

Evaluation of feeds for energy for ruminants and Non-ruminants Evaluation of feeds is concerned with the assessment of the quantities in which nutrients are supplied by feeds as well as the assessment of the quantities in which they are required by different classes of farm animals. The potential value of a feed to supply nutrients is known by chemical analysis whereas the actual value is calculated by making allowances for the inevitable losses that occur during digestion, absorption and metabolism. The major losses occur through feaces. The major organic nutrients i.e. energy and protein are required by animals for construction of body tissues, synthesis of milk, meat and eggs. All these functions involve transfer of energy from chemical to heat energy (through oxidation of nutrient) or from one chemical form to another (carbohydrate is used for body fat synthesis). Evaluation of feed for energy involves fate of feed energy in animal body, measurement of energy metabolism and expression of the energy value of feeds. Proteins are made of essential (indispensable) and non-essential (dispensable) amino acids, which should be in sufficient amount to meet the metabolic demands for maximum efficient feed utilization. Monogastric animals get amino acids from breakdown of feed proteins during digestion and absorption but ruminants get amino acids from proteins synthesized by the microbes in the rumen as well. Basic terms Calorie (cal)- A calorie is the amount of heat required to raise the temperature of 1 g of water from 14.5 oC to 15.5oC. As the specific heat of water changes with temperature, a calorie is 4.184 internatioanl joules. Kilo calorie (Kcal) A kilo calorie is the heat required to raise the temperature of 1 kg of water by 1oC. A kilo calorie is equal to 1000 calories. Mega calories (Mcal)- A mega calories is equivalent to 1000 K cal or therm. International union of nutritional sciences (IUNS) has recommended the calorie concept. British thermal unit (BTU)- A BTU is the amount of heat required to raise 1 lb of water by 1oF. A kilo calories approx. equal to 4 BTU. In UK the kilo joule is used. As per international system of units (SI) the unit of energy is the joule (J) 1 kilo calorie 4.184 KJ 1 KJ= 0.239 Kcal The quantity of energy present in feed is calculated by converting chemical energy to heat energy and determining the heat produced. Factors affecting conversion of ME to NE 1. Nature of the feed Feed high in fiber or protein need more energy for digestion and metabolism. The energy needs of fiber > protein >starch > fats. Thus feed fiber in winter and fat rich diet in summer.

30

2. 3.

Purpose for which it is used- Energy used for maintenance is less than for production. Hence the efficiency for maintenance is 80%, for growth 70% and for lactation 72%. The level of production- The higher the level of production the better is the use of ME.

Calculate the NE of roughage for maintenance DMI = ME = Heat production maintenance = Heat production fasting condition = HI = 2013 Kcal NE = ME-HI/DMI

3.79kg 10508 Kcal 9803 Kcal 7790 Kcal

= 10508-2013/3.79 = 2241 Kcal/kg DM

Direct and Indirect Calorimetry To find the ME utilized by the animal It is necessary to measure the animals heat production (HP) or its energy retention (NEp/NEg). NE NEp/NEg NE = ME-HI HP = HI + NEm NEp = ME-HP Heat production is due to oxidation of food and oxidation of body fat and body protein. Heat production can be measured by animal calorimeter which is known as direct calorimetery. Heat production can be made by estimating the respiratory exchange of gases using a respiratory chamber. This is known as indirect calorimetery. Respiratory chamber can also be used to measure energy retention by estimating the carbon and nitrogen balance. Lavoisier was first to recognize that heat is produced by oxidation of food. Lavoisier and Laplace (1780) devised the animal calorimeter. An adiabatic calorimeter uses a chamber of water to measure the heat given off by the animal. Now, gradient layer calorimeter is used where the gradient of temperature between the inner and outer layer is used to calculate the heat given off. The respiratory calorimeter has both the respiratory chamber and a calorimeter. It is presumed that the heat lost from the body is equal to the heat produced. The heat is lost from the body is as radiation, conduction and convection (sensible heat loss) and by 31 NEm

evaporation of water from the skin and lungs (evaporative heat loss). The animal calorimeter is an air tight insulated chamber where the heat lost by the body increases the temperature of the circulating water. The rate of flow of water and the change of temperature is used to calculate the heat lost. In addition to the metabolic trial the CO2 and methane loss are accounted for. The HI is estimated as the difference in the heat produced at two levels of intake. Two levels are taken to account for the basal metabolism which remains the same. The extra heat produced at higher level of intake is due to the higher feed intake. The procedure for estimating energy by this method is very expensive. Indirect calorimetry by the measurement of respiratory exchange (Respiratory Quotient) Carbohydrates, fats and proteins are oxidized to give energy. For carbohydrates it is : C6H12O6 6 CO2 + 6 H2O H (675 Kcal /mole) Oxidation of fat (stearic acid) is: CH3 (CH2)16 COOH + 26 O2 H (2711 Kcal /mole) 18 CO2 + 18 H2O -

1gm molecule of oxygen occupies 22.4 L at NTP. So energy obtained solely from glucose is 675 4.2 6 22.4 = 20.98 KJ (5.014Kcal) of heat. This value is known as thermal equivalent of oxygen. The thermal equivalent of oxygen varies with the nutrients and their mixtures. Respiratory quotient: This is the ratio of volume of CO2 produced to volume of O2 used. Under same temperature and pressure the same volume of gases contains same number of molecules. So. RQ of carbohydrate is 1 (6O2 and 6 CO2) and fat is 18 CO2/26 O2= 0.692 and for protein is 0.8. Metabolic body size- It is the weight of the animal, in kg raised to three fourth power (Wkg 0.75). It may be termed as physiological weight. The body weight is the physical weight (gravitational weight).

32

Proteins value measurement and utilization Protein: Greek word proteios means first. What is a protein: Proteins are giant polymeric compounds made of amino acids, linked together with peptide bond (NH-C- ). More than 200 amino acids are known. Only 20 are commonly found in most proteins. In human organism, 100000 different proteins are there. In 1.5 million sp. there are 10
10

-10

12

different kinds of protein

molecules. Proteins contain C 15%, H 7%, O 23%, N 16%, S 1-3% and P <1%. Crude Protein: Most of the dietary nitrogen required in the body is used for protein synthesis. Crude protein of a feed constitutes true protein and the non-protein nitrogen (NPN). The NPN part of the proteins may or may not be useful for all the spp. of animals. The NPN constitutes free amino acids, amides (asparagines, glutamine, urea) amines (betaine, histamine, tryptamine, tyramine) alkaloids (cocaine, morphine, nicotine, solanine, strychnine), ammonia, nitrates, nucleic acids (RNA, DNA) purines, pyrimidines, uric acid. Chemically the crude protein content of a food is calculated from its nitrogen content after its estimation by kjeldahl method. There are two assumptions for calculations of the CP content of food. Assumptions 1. 2. All the protein contain 16% N in them. All the nitrogen present in the food is I the form of protein.

CP% = %N 100/16 = % N 6.25 Both these assumptions are not sound since all the proteins have different nitrogen content in them. ______________________________________________________________ Type of feed Mature seeds Leaves Stems and roots Storage organs % N as true protein 95 80-90 60 30-40 33

Since the N content of all the proteins are different thus their conversion factors from N to CP content are different. Ingredient N% Conversion factor (%CP=%N factor)

Cereals Maize Wheat Oats Barley Oilseeds meals Soybean Cottonseed Animal feeds Egg Meat Milk 16 16 15.68 6.25 6.25 6.38 17.51 18.87 5.71 5.3 16 17.2 17.2 17.2 6.25 5.83 5.83 5.83

Though the feeds contain a variety of NPN compounds, quantitatively only amides and amino acids are important. Amino acids: Amino acids are hydrocarbons containing one or more than one basic (NH2) group attached at group and one carboxylic (COOH_ group. Depending upon the metabolic needs the amino acids are classified as essential and 1 essential amino acids. Metabolically 22 amino acids are essential but some of them are dietary essential. Essential amino acids: Those amino acids which may or may not be synthesized in the body if synthesized that too only to a limited extent which cannot meet the metabolic requirement of the animal/bird are called essential amino acids. These are A V H I L L M P T T or Arginine, Valine, Histidine, Isoleucine, Leusine, Lysine, Methionine, Phenyl alanine, Tryptophan and Threonine. The other amino acids are made available in the tissues from the transformation of other amino acid available.

34

Properties of proteins: They are colorless, crystalline generally soluble in water, slightly soluble in alcohol and insoluble in ether. Due to the presence of both acidic and basic groups these are readily ionized. Amino acids behave as Amphoteric electrolytes and give both anions and cations in solution. On hydrolysis of the proteins in the intestine of monogasrics the proteins liberate constituent amino acids. These amino acids are absorbed from the intestinal lumen and are available to the system. For the maximum efficiency the animal/birds must receive the correct quantity of all the essential amino acids in the proper proportion along with the sufficient quantity of non-essential amino acids. True protein: True proteins of a feed can be separated from the NPN compounds by precipitation with alkaline cupric hydroxide or in some pant material by heat coagulation. The true proteins are filtered and the residue is subjected to N determination by using kjeldahl method. True protein = % N of residue 6.25 Digestible crude protein : The values of CP of feeds are meaningless since he constituent protein undergo hydrolysis in the intestine to give amino acids before there are made available to the system. The DCP content of a feed can be calculated as follows. DCP = CP content of the feed % digestibility coefficient. Different proteins have different digestibilities in the carious Sps. Thus the DCP values of a feed for various spps. are different. DCP is used fro ruminants only. The DCP of protein = N in the feed N in the feaces/ N in the feed 100 This digestibility coefficient of CP is apparent i.e. it is not true. Since the feacal N contains nitrogen from two origins. 1. 2. Dietary origin undigested dietary protein Metabolic origin : enzymes, desquamated cells

35

In rumen the NH3 produced in the rumen may be absorbed across the rumen wall. This NH3-N is also considered to be digested. Thus the DCP values may be misleading. The true digestibility of CP can be calculated by considering the metabolic N losses. True digestion coefficient of CP = N in the feed (N in the feaces + N of metabolic origin)/ N in the feed 100 The in vitro digestion of proteins can also be determined by incubating the proteins in the incubator at 37oC with pepsin. This method is limited because only one enzyme is considered. Results also vary with 1. 2. 3. Fineness of grinding Concentration of enzymes Conditions of drying the feed

It may be used as comparative evaluation. MFN in pigs = 6g protein /kg DM In Ruminants = 30g protein / kg DM (high due to microbial residues and low protein in feeds) Measurements of protein quality for monogastrics The crude protein, true protein and DCP values are not entirely satisfactory. Moreover, the efficiency with which the absorbed protein is used differs considerably from source to source. In order to take this in account, methods based on the animal responses have been devised. 1. Biological value: The term biological value (BV) is defined as the percent of absorbed nitrogen retained by the body for repair or synthesis of nitrogenous tissue. It involves the determination of the nitrogen in feaces and urine when known quantity of the protein under test is fed.

36

BV = 100 N intake (Feacal N-metabolic N) (Urinary N- endogenous N)/ N intake - ( Feacal N-metabolic N) The method demands that the nitrogen in diet should not be in excess of the amount needed for maximum growth as at high level the excess protein will be catabolized and this will lower the BV. Besides this the diet must be adequate in non-protein and other nutrients. Limitations: 1. It is difficult to select the optimum level of protein to be fed to obtain the true BV. 2. The feaces and urine needs to be collected 3. The method is time and labor intensive. BV of some feeds: Barley- 57-71, wheat- 67, Corn- 60, Soybean- 62-65, Peas- 63-76, fish meal- 74-89, egg- 94, milk- 95-97, cottonseed meal- 63, linseed- 61. Calculation of BV FI- 6.0g, %N in feed 1.043 NI/d = 62.6 mg, UN= 32.8 mg, EUN= 22 mg, FN= 20.9 and MFN 10.7mg BV= 62.6-(20.9-10.7)-(32.8-22)/62.6-(20.9-10.7) 100 = 79

2. Protein Efficiency Ratio (PER): ratio of weight gain and protein consumed by the animals/birds. PER = weight gain/protein consumed. For PER records of growth rate for a period of time and the feed consumed/ protein consumed is maintained. The values of PER are affected by age, sex of the

37

animal/bird, length of feeding period and level of protein in the diet. Usually a four week period is taken in case of rats PER values are frequently stated relative to the standard casein with an assigned PER values. Limitation: 1. 2. 3. 4. Very tedious Cannot asses the protein digestibility Cannot asses the proportion of protein consumed used for the maintenance and or tissue growth The weight gain recorded may be due to accumulation of water, bone formation or fat deposition. Thus protein content of weight gain is very variable. 3. Net protein retention (NPR): It is a modification of PER method which takes care of the partitioning of protein consumed in maintenance. Thus it is considered to provide more accurate results than PER. In this method weight gain of experimental animals/ birds is compared with a group maintained on a protein free diet. NPR = weight gain on TPG weight loss of NPG/ protein intake Where NPR = Net protein retention TPG = Group fed on test protein NPG = Group maintained on non-protein diet (protein free diet) 3. Gross Protein Value (GPV): In this method relative ratio of weight gain per unit of test protein to that of a standard percent protein consumed is taken. Chicks are fed diet with 8% CP for 2 weeks and then divided into three groups I II III Basal diet with 8% CP Basal diet + 3% CP for test protein Basal diet + 3% CP from casein

GPV = (g increased wt gain/g test protein)/ (g increased wt gain/g casein)

38

The extra live weight gain per unit of supplementary test protein, stated as a proportion of extra live weight gain per unit of supplementary casein is the GPV of test protein. The different methods of evaluating nutritive value of proteins are placed into two main headings: A B Biological methods Laboratory methods

A Biological methods: These involve experimental animals and the protein to be tested is fd as sole source of protein or as supplement to cereals. These methods are a) Gain in weight: This measures the rate of growth over a particular The method period of time to record the nutritive value of the protein. The advantage of this method lies in its simplicity. The limitations are: 1. can be due to deposition of fat or water. 2. 3. b) The same gain in weight by two different protein sources may be Time consuming Rate repletion method: This method is based on the principle common to many bioassays i.e. the production of deficit followed by a measurement of the replacement of that deficit. Depletion can be accomplished by feeding protein free diet until the rats loose 25% of their initial body weight. The animals are then fed nitrogen in the test diet and the rate of repletion is measured. Merits: 1. 2. It eliminates the effect of palatability and unequal protein intake. Relatively shorter period required. different i.e. it may be gain in protein, fat or bones etc. assumes that the gain in eight is due to growth but many times the weight gain

39

3. 4.

Sample required is less The same animal can be used several times with a gap of 10 days between tests.

This method creates artificial conditions and the results may not reflect the quality under normal conditions. c) Protein minimum or nitrogen balance: The method is based on minimum amount of protein required to maintain the nitrogen equilibrium. The data is obtained by actual feeding of the test protein and determining the N-balance. The minimum N is estimated by plotting N to the point of exact equilibrium. Limitation: 1. The results of this method are very variable. Protein minimal was low in a protein depleted animal and high in animal with full protein stores. 2. The animals need to be standardized to similar steady states when one protein minimal is compared with another. e) Nitrogen balance index: also called Net Protein Value. This method uses relationship between 1. 2. 1st relationship The relation between NB and absorbed N is given as B = K(A)-Bo -----1 where Bo= sun of urinary nitrogen and feacal Nitrogen at zero nitrogen absorption or metabolic nitrogen A = absorbed nitrogen K = rate of change of nitrogen balance w.r.t. absorbed nitrogen and is called nitrogen balance index. Nitrogen balance (B) and absorbed N (A) Nitrogen intake (I) and nitrogen balance (B)

40

Or K = B+ Bo/A 2nd relationship Between nitrogen intake I and nitrogen balance B B = K (I) Bo ----2 From 1 and 2 KA-Bo= K(I) Bo = B Or KA = KI K/K = A/I Or K/K = D Where D is the digestibility K function of digestibility retention of body and diet nitrogen. The K has the same significance as Net Protein Value which is BV DC. Thus it is the overall utilization of dietary protein by the animal. If K and K are same then NPU is 1 which means all the protein fed is digested and absorbed completely Limitations: a) a protein may yield a high BV or nitrogen balance index for absorbed nitrogen (K) but may have low index for intake because of poor digestibility. b) Collection of feaces and urine is tedious. f) Protein replacement value: As the maintenance of animal on a protein free diet is difficult the protein replacement value was introduced (PRV) which is given as PRV = NB1-NB2/NI 100 Where NB1 is nitrogen balance of the animal fed reference protein, NB2 is nitrogen balance of its pair-mate fed the protein under test and I is nitrogen intake in pair.

41

So one group is given reference protein diet and other test protein diet and the difference between the two is the PRV. Limitation: a) while it removes the limitation of BV it still has the drawback of other methods. It does not show any improvement of protein. Calculations NB1 of egg is +20 and NB2 of soybean is +16 and nitrogen intake (NI) = 50 mg so PRV = 20-16/50 100 = 8% that is soybean has egg replacement value of 100-8= 92%. h) NPU or body N-retention method: The method involves the estimation of total nitrogen content of the carcass of test animals. There are two groups of chicks one fed at low protein level and other at 10% higher protein level for 10 days and the gain in body nitrogen is calculated as NPU = Body N of test group Body N of non (low) protein group + N consumed by non (low) protein group _______________________________________________________ N consumed by test group It was further simplified by predetermination of water: N ratio based on the claim that N: water of the body is a constant ratio when measured on a fat free basis. The advantage is that the time period required is less and there is direct body N determination. Limitations: 1. 2. 3. The animal carcass is analyzed so it can be used only for one test There is no additional advantage over others The N content of control group determined in the beginning is on the

assumption that nitrogen content in all the animals is the same.

42

i)

Liver nitrogen content: In this the sensitivity of N content of the liver to Limitations: 1. The limitations are the same as in NPU. 2. It is assumed that the liver N content of control group at the end of experiment is assumed to be the same in all the animals.

different dietary proteins is used for determining the nutritive value of the protein.

j)

Enzymatic activity in liver: It is based on the assumption that protein loss from

tissues through depletion results in the loss in enzyme activity. During fasting there is a reduction in the activity of enzymes catalase, alkaline phosphatase, xanthine dehydrogenase in the liver and the loss in activity parallels or exceed the reduction in liver protein. Limitations: Same as in NPU. Proteins are not the only component to affect enzyme activity which is also affected by the vitamins especially B-complex which act as cofactors. k) Changes in blood proteins: The protein depletion and repletion results in rise and fall in plasma albumin and other proteins ( and globulins) and amino acids. PAAR = Av. raised AA level in plasma Fasting level of amino acid/ Percent amino acid required in the diet Where PAAR is Plasma amino acid ratio Limitations: 1. The reduction in plasma amino acid often results in reduction in plasma volume. The concentration effect in plasma water may sometimes not decrease albumin and may even increase globulin. 2. Estimation of amino acids or albumin is not possible in all laboratories.

43

l)

Creatinine : Nitrogen ratio in urine: It is based on the findings that the

creatinine excreted in urine is constant regardless of the diet and more nitrogen is excreted from poor quality than good quality protein. It is a simple and rapid method of estimation. Limitation: The ratio lacks uniformity due to the variation in creatinine excretion by animals of different ages. m) Gross protein value: A low level of test protein (3% protein) is fed as a supplement to a higher level of cereal protein (8%). Chick growth is measured foe 14 days and a growth on test protein is determined by subtracting the weight gains of birds fed only on cereal containing diet. The results are expressed as a percentage of the value obtained when casein is used as the test material. The method is better as it simulates the practical feeding. Limitations: 1. 2. 3. B) The combination of cereals is arbitrary ad the GPV may vary with the The results at 11% protein level may not be repeatable at 20% protein Since cereals are deficient in lysine the results may essentially measure the Laboratory methods: Since the biological methods are variable, time composition of the cereals

contribution of lysine in the protein supplement consuming and uneconomical; laboratory methods have been evolved to measure the nutritive value of proteins. a) Hydroxyproline content: This method is specific for animal proteins. The animal proteins contain connective tissues which contain collagen which has lower nutritive value as compared to muscle protein. The hydroxyproline is present in the collagen and it is made water soluble by boiling in water. b) Protein quality index: When four fractions of nitrogenous material; copper

precipitable, pepsin indigestible, hot water soluble and phosphotungstic acid soluble

44

are combine it gives an index called protein quality index. The results of PQI are very variable. c) Chemical score: In this the amino acid composition of the test protein is

compared with that of egg protein and the most limiting amino acid is expressed as percent of the same amino acid in egg protein. Limitations: 1. 2. 3. It is expensive and time consuming It is based on the assumption that egg protein fully satisfies the animals requirements The amino acid composition of heat treated protein is not a true representation of its amino acid availability.

Example: Find the chemical score of wheat protein Lysine is most deficient and is 63% deficit. Therefore the chemical score for wheat is 100-63=37% Lysine in egg is 7.2% of CP and in wheat it is 2.7% of CP so chemical score is 27/72 100 = 37 e) Essential amino acid index (EAAI): Since other amino acids are also

important this method considers 10 amino essential acids. It is defined as a geometric mean of egg ratio given by Egg ratio = A.A. in the test protein/A.A. in the egg 100 This egg ratio for 10 amino acids is calculated as EAAI = 10 (1+2+3+--------------+10)

45

Advantage: 1. 2. It provides a better result of nutritive value than chemical score it can estimate the effect of fortification of amino acids or proteins

Limitations: The limitations are the same as in chemical score. f) Pepsin digestibility: In this method the protein is digested with pepsin.

Although the method is easy it presumes that all the digestible protein is available which may not be true. g) Available lysine value: Lysine is the most limiting amino acid and also it is

most likely to be affected by heating protein. It involves refluxing the protein to react the epsilon amino acid group of lysine with 1 floro 2,4 dinitrobenzone (FDNB) for 16 hrs. The amount of free amino group is determined calorimetrically. It is good for animal proteins but not vegetable proteins. It does not take into account the other essential amino acids. h) Microbiological assay: In this microbes are used to determine the nutritive

value of proteins by measuring the growth or turbidity of acid production by the microbes. eg. Streptococcus zymogenes which gives good result for fish meal, and a ciliate protozoa Tetrahymena phriformis Ileal digestibility: Digestibility coefficients based on collection and analysis of digesta from ileum (terminal point) are generally considered to give a more accurate measure of N-absorption than do the feacal collection method. Bacteria present in large intestine of pigs, poultry, dogs and cats change the amino acid composition of undigested food residues. These bacteria both add and consume amino acids. Thus feacal material contains undigested and bacterial amino acids. Thus it gives better and accurate determination of nutrients. Ileal collection eliminates the error due to large intestines.

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Protein absorption from large intestines makes little or no contribution to the protein of the animal. Ideal protein concept: The most recent method of evaluating the protein for growing pigs and poultry birds is based on ideal protein. It is a modification of chemical score with amino acid pattern of the particular tissue protein serving as reference pattern. If the main limiting amino acid was lysine at 50 g/kg CP then the score would have been 50/70= 0.35 and the ileal protein content would be 700 g/kg CP. A diet with 170 g/kg of this protein would then supply 170 0.7 = 119 g ideal protein /kg. The requirement of any indispensable amino acid for any rate of growth has a fixed proportion to the other in the diet. The concept of ideal protein was originally proposed by Mitchell in 1946 to match exactly the amino acid requirements of the animals for growth and maintenance thus avoiding over and under feding of amino acids. During 1981 ARC received the idea and published the 1st estimate of ideal protein requirement of growing pigs as under Recommended valance of amino acids (g/kg) in ideal protein _____________________________________________________________ Lysine Meth + Cystinea Threonine Tryptophan Isoleucine 70 35 42 10 38 Leucine Histidine Phenyl alanine + Tyrosineb Valine Non-essential amino acids 70 23 67 49 596

a: At least half should be methionine b: At least half should be phenyl alanine The benefit of ideal protein is to sell the requirements of all the essential amino acids on the basis of lysine. So it is a statement of EAA requirements in a proportional

47

relationship to the requirement of lysine. Thus the dietary concentration of lysine is set and the concentration of all other EAAs is determined as a percentage of the lysine concentration according to ideal amino acid concentration. For poultry the evaluation of protein sources is based on their contents of three major limiting amino acids i.e. lysine, methionine and tryptophan. It is assumed that diets sufficient in these three amino acids will automatically get sufficient amount of other amino acids. Measures of protein quality for ruminants The quality of protein is not important in case of low producing animals since all the essential amino acids are synthesized by the rumen microbes. However, it is important for high producing animals and a small portion of by-pass protein is advocated to meet their requirements for higher growth and milk. 1. 2. CP: Contains variable amount of NPN compounds so it is unsatisfactory because no allowance was made for the nutritive value of NPN fractions. Protein equivalent: In an attempt to have NPN fraction half the nutritive value of true protein a concept of protein equivalent (PE) was introduced PE= %DCP + % DTP/ 2 3. DCP: There is dissatisfaction for the use of DCP for evaluating feed proteins due to extensive degradation and synthetic activity of rumen microbes. Its use was rejected because 1. Dietary proteins are largely degraded in the rumen 2. The extent of degraded N is utilized by the microbes in relation to the amount of energy fermented. 3. Feacal excretion could be altered by manipulating the site of fermentation between rumen and caecum because of easy calculations it is widely used. 4. Biological value: It does not apply to ruminants

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5. 6. 7.

Metabolizable proteins: System was used in USA Rumen degradable and undegradable protein was used in UK True protein digested in small intestines (PDI) in France The fundamental principle underlying this system is that N requirement is

considered as under: 1. 2. Requirement of N for microbes Requirement for the host ruminant

The protein value may be assessed from the determined or estimated flow of protein or amino acids leaving the rumen or entering the small intestine. Intestinal supply is from 1. 2. Dietary proteins escaping rumen degradation Microbial proteins due to fermentation in the rumen, sloughed cells and secretions in abomasums. Metabolizable protein: It is that part of the dietary protein and microbial protein which is absorbed by the host animal and is for use at tissue level. In vivo and In vitro methods have been employed to estimate either the total protein supply or individual component. ARC (1980) adopted microbial protein yield as 30 g N/kg digestible OM. It can be estimated by using rumen cannualated animals with the help of markers such as amino ethylophosphoric acid (AEPA (protozoal marker) and diaminopimelic acid (DAPA) . Protein flow to small intestine can be determined with animals equipped with simple or re-entrant cannulae to the abomasums or duodenum. Rumen degradability: (N/CP) is estimated by In sacco /In situ technique to know RDP and undegradable protein UDP.

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Hay making High quality hay is green in color, leafy and pliable and free from mustiness. Indian hay is of poor quality as it consists of dry grass on which the seed has ripened and leaves have usually been shed. In feeding value it is similar to straw of cereals. During monsoon season when grass is ready to be cut fro hay the weather is often wet and after monsoons grass mature and cultivars are busy making preparation for rabi sowing. Principles of hay making: It is to reduce the moisture content to a safe limit to be stored without undergoing fermentation or becoming moldy. The hay should not have been leached by rain while making and loss of leaves be kept minimum. Hay becomes handy when fodder availability is scarce or when fodder is very succulent and addition of dry fodder is necessary. Requisites of good quality hay: 1. It should be leafy. Leaves are rich in proteins, vitamins and minerals. Loss of leaves during hay making means loss of valuable nutrients and final product of less feeding value. 2. It should be harvested at the right time (Milk stage or at the flowering stage, crop is in bloom).Delayed cutting results in nutrient being diverted to seed making. 3. It should be green in color (rich in vitamin A precursor) 4. it should be soft and pliable. 5. It should be free from dust and moulds. 6. It should be free from weeds and stubbles. 7. It should have the smell and aroma characteristics of the crop. 8. Moisture should not exceed 15%. 9. It normally has 25-30% CF and 45-60% TDN. Kinds of Hay 1. Legume hay: They have higher TDN, CP protein of good quality, vitamins especially carotene a precursor of vitamin A. It may have vitamin D and E. It is rich in Ca, palatable. These hay are made from Lucerne, berseem, cowpea, soybean. 2. 2. Non-legume hay:- made from grasses they are inferior in quality. These are less palatable , have less proteins, minerals and vitamins but grass production is more than legume and it can be grown easily. Hay of oats and barley compare favorably with other grass hays., If harvested at the proper stage and made into hay they are rich in carbohydrates although they are lower in proteins and minerals in comparison to legume hay. Oat is a good non-legume crop for hay making. 3. 3. Mixed hay:- hay made from mixed crop legume+non-legume. Such crop are generally cut earlier due to variation in the seeding time. If harvested early, cereals are richer in proteins. Harvesting of crop for hay making 50

It should be harvested during day time after dew has dried off. The field should not be wet otherwise the drying will not be uniform. Curing of hay During curing of the hay should be saved from bleaching by the sun and as far as possible the leaves should be preserved from shattering. It is best if crop is left in the field for a few hours until wilted or about 1/4 th to 1/3rd cured. It should be raked into small loose heaps. If weather is wet then turn over the heap to hasten curing. It can also be hanged on fences or in oven. To avoid excessive loss of leaves it is better to handle the hay early in the morning. Moisture should not exceed 1520% or else fermentation will occur, heat produced causing loss of nutrients. Technique:- Berseem hay:- Chop berseem/ Lucerne in pre-bloom or bud stage. Spread into thin layers 4-5 inches layer. Stir for drying store on drying. Losses of nutrients in hay making 1. Losses due to late/early cutting:- if late then greater lignification, lower carbohydrates and protein digestibility. If early low yield and high moisture. 2. Losses by shattering:- losses by laves shattering reduces its nutritive value. (They are high in TDN, proteins, vitamins and minerals) 3. Losses of vitamins:- During prolonged curing or severe sunlight carotene is lost due to bleaching. Extent of bleaching is proportional to the greenness of hay. 4. Losses in fermentation:- During fermentation of hay starch and sugars are oxidized to CO2 and H2O. If drying is prolonged mould grows which makes the hay unpalatable and may produce Mycotoxins. Atinomycetes growth on hay can cause allergic condition called farmers lungs. 5. Losses by leaching:- Hay when prepared if left in the field would loose nutrients due to leaching. Leaching occurs only if the hay is soaked in water but to a lesser extent if it is there in a heap. Biochemical changes during hay making and storage Soon after harvest the fodder starts wilting but before most of the moisture is lost the plant respiratory enzymes activity continue resulting in oxidation of valuable nutrients. The process also continues during storage if there is sufficient moisture. 1. Carbohydrates:- The water soluble carbohydrates are oxidized to CO2 and H2O with production of heat and loss of dry matter. Losses occur of glucose, fructose, malic, citric and succinic acids. 2. Nitrogenous constituents:- Losses of nitrogenous substances is insignificant. There is increase in amino acids due to proteases activity. The toxicity of jowar reduces due to denaturation of the enzyme responsible for converting cyanogenic glycosides to HCN. 3. vitamins:- Slow drying in the sun causes 80% loss of vitamin A due to the action of lipoxidase on carotene. Quick drying reduces the losses due to rapid inactivation of the enzymes. Exposure of ultraviolet rays of sun on ergosterol converts it into ergocalciferol therby increasing Vitamin D.

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Mature forages are lower and dryed hay still lower in Vitamin E activity may be due to low leaf:stem ratio.

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Silage making What is Silage: Silage is a fermented feed resulting from the storage of high moisture crops, green forages, under anaerobic conditions in a structure known as a silo. Ensiling or Ensilage: Refer to the chemical and physical changes that take place when feed or forage with sufficient moisture are stored in a silo in the absence of air. Silo: It is an airtight to semi-airtight structures designed for the storage and preservation of silage. Size and Characteristics of a silo pit: 1. The silage should be decided on the basis of the i. number and type of animals to be fed daily ii. ii. The length of the feeding period and the iii. iii. Amount of forage available for ensiling. 2. The side walls should be straight and smooth in order to prevent the formation of air pockets to which may retard the normal microbial fermentation. 3. It should have adequate depth, thereby making fro better packing and less surface area to total mass exposed. 4. The walls should be strong and rigid in order to withstand the pressure which develops inside the pit as fermentation takes place. 5. Adequate provision should be made fro the escape of surplus juices by a drain on ground bottom. 6. It should be conveniently located to easy access, filling and feeding. 7. It should be at an elevated spot on the farm to avoid water seepage. 8. The depth should depend on the water table of locality. Kinds of crops for silage: As crop having sufficient soluble carbohydrates and moisture to produce the desired quantities of acids may be used. The most commonly used crops are maize, sorghum, sudan grass, bajra, napier etc. of these maize and sorghum are the best crops. In leguminous crops Lucerne, berseem and cowpeas are used. They are not good cops for silage making but can be used after treatment. This is so as they are low in soluble sugars so a solution of molasses is sprinkled at every 1/3 meter of filling. Preparation of forage for making silage: The crop to be used for silage making should be harvested a the proper stage of maturity, cut to proper length, of adequate moisture content, filled rapidly and distributed uniformly in the silo. After filling properly, trampling should be done properly with the help of men, buffaloes or tractor. Fill the pit 3-4 ft above the ground level. Cover from all side with long paddy stem/grass upto 4-5 ft and cover with mud to seal the pits. The silage is ready in 2 months after covering. 53

Proper stage of maturity: The crops are generally harvested at the flowering stage when it has maximum nutrients. Maize early dent stage, sorghum- late dough stage. Silage with less than 25% DM makes very good silage and with significant amount of silage juices containing nutrients. Recommended dry matter for silage making is 35-40%. Cut to proper length: The length of the cut sections affects its packing and silage quality. It varies from an inch to over an inch for grasses, mixed and dry forages and forages with hollow stems require finer chopping. Moisture: about 60-65% moisture and 35-40% DM is best for most crops. Higher moisture in crops results in slimy and putrefied silage due to presence of butyric and undesirable acids. They will be excessive seepage of juices an hence loss of nutrients. There will be excessive deterioration of moisture crops. Adding ground grains eg. Maize or cob meal, brans helps. Additives or Preservatives in silage preparation i. Molasses: some forage like legumes are low in sugar content. Addition of molasses increases lactic acid and acetic acid content and improves the quality of silage. It is added at the rate of 3.5% may be upto 5% of green weight of forages. ii. Urea: urea at the rate of 0.5% to 1% of fresh forages with the purpose of improving nitrogen level of forages. iii. Limestone: this is calcium carbonate added @ 0.5-1% to neutralize the acid so that lactic acid bacteria can perform longer and produce more lactic acid. iv. Sodium metabisulphite: SO2 gas evolved from it acts as anti bacterial preservative and improves carotene content. v. Organic acids: Propionic and formic acids are used as preservatives. vi. Bacterial cultures: cultures of acid forming bacteria Lactobacillus acidophilus are added as innoculum to increase the number of bacteria and for rapid fermentation common salt 0.5% to improve the palatability. Filling of forage should be rapid and completed within 2 days. The forages should be compressed simultaneously to prevent the presence of air pockets. Finally the top of the silo should be covered by an insulator like plastic or loose earth. How silage is formed: When the forage is chopped and packed in a silo the cells continue to respire and use the O2 present in the silo and give out CO2. In 4-5 hours the O2 is used and CO2 increases. After 48 hours the CO2 level decreases with the production of other gases like CH4, CO, NO2 etc. this condition promotes organic acids production by bacteria. Eventually the acids kill the bacteria and preserve the silage. The plant forage carries a lot of aerobic fungi and bacteria. As conditions turn anaerobic the microorganisms are killed. This situation is further favored by rise of temperature to 80-100oF (27-38oC) in about 15 days. Later the temperature reduces. 54

During fermentation sugars are changed to organic acids and some proteins to amino acids. These are utilized by ruminants. But in badly preserved silage the amino acids are converted to amines eg. histamines, phenyl ethyl amines etc. which at higher concentration proves harmful to the animals. Chemical changes Homo-fermentative lactic acid bacteria Glucose, fructose + 2 ADP Pentose 2 Lactic acid + ATP Lactic acid + acetic acid

Hetro fermentative bacteria 1 glucose Lactic acid + 1 ethanol + CO2 + ATP 3 fructose 1 Pentose Clostridia Alanine + glycine 3 Alanine Arginine Histidien Leucine Lysine Phenylalanine Tyrosine Tryptophan 1 Lactic acid + 2 mannitol + 1 acetic acid + CO2 + H2O 1 Lactic acid + 1 acetic acid 3 acetic acid + 3 NH3 + 1 CO2 2 propionic acid + 1 acetic acid Putrescine Histamine iso veleric acid cadeverine Phenyl ethyl amine Tyramine Tryptamine

Characteristics of a good silage: 1. Very good silage: Clean, acidic to taste, has no mould growth and butyric acid. The ammonical nitrogen should be less than 10% of total nitrogen. It is green or brownish in colour pleasing taste not bitter or sharp. pH 3.5 to 4.2 with average of 4.0. 55

2. Good silage: Acidic to taste, has traces of butyric acid, pH between 4.2-4.5, ammonical nitrogen 10-15% of total nitrogen and other points same as very good silage. 3. Fair silage: It has a little amount of butyric acid. There is slight proteolysis with some mould growth, pH between 4.5-4.8, ammonical nitrogen 15-20% of total nitrogen, colour is tobacco brown to dark brown. 4. Poor silage: It gives a bad smell due to high level of butyric acid and high proteolysis. The silage may be infested with moulds, pH above 4.8, ammonical nitrogen is more than 20% of total nitrogen and colour is blackish. Advantages and disadvantages of silage Advantages 1. Green fodders can be kept in succulent condition for a long period and used during summer 2. Silage preserve about 85% of nutrients 3. Whole stalks of maize or sorghum can be fed but not when dried so it is more economical 4. During monsoon hay cannot be made but silage can be made 5. Hard crops can be used for silage making as the weeds are harvested before seed formation so I is a way to control weeds in fields. 6. It is palatable and laxative in nature 7. It preserves proteins and vitamins especially carotene 8. The field is free for preparation of next crop 9. Silage occupy less space than dry fodder (1/3 space as compared to hay) 10. Crops can be harvested at optimum maturity so more nutrients are available Disadvantages 1. It requires a silo 2. It possess less vitamin D as compared to sun cured hay 3. Extra labour is needed for filling the silo.

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Use of urea in animal feeding Feedstuffs that contain nitrogen in a form other than proteins or peptides are termed non-protein nitrogen (NPN). The NPN compounds can be either organic or inorganic. Organic NPN includes ammonia, amides, amines and amino acids. Inorganic NPN includes a variety of ammonium salts and ammoniated byproducts. Some NPN for ruminants N% Protein equivalent% Ammonium acetate 18 112 Ammonium bicarbonate 18 112 Ammonium carbamate 36 225 Ammonium lactate 13 81 Biuret 35 219 Urea feed grade 42-45 262-281 Urea pure 46.7 292 The av. nitrogen content of urea is 45% with protein equivalent of 281%. The protein equivalent = 6.25* %N. Microorganisms in the rumen degrade the dietary protein and are able to to synthesize their own body protein if readily available carbohydrates are available. Of the various NPN source urea is the most important. H O H N-C-N H H Starch VFA

urease Urea NH3 + CO2 In rumen Microbial proteins Utilization of urea in the rumen: Urea in the rumen is broken down into ammonia and carbon dioxide by a microbial enzyme urease. Side by side other microbes degrade starch to produce acetic acid, propionic acid or butyric acid which is used as energy sources. In the multiplication and growth the rumen microbes used the ammonia released from the breakdown of protein and non protein and NPN compounds, along with the carbon skeleton and reduced sulphur to form amino acids which re in turn manufactured into microbial proteins. Some of the unutilized ammonia is absorbed across the rumen wall and is detoxified by the liver to give back urea from excretion or re-circulation. 57

Urea-fermentation-potential(UFP): It is a term used to indicate the amount of urea that can be utilized in a ruminant ration. A positive UFP value of the feed/ration can be defined as grams of urea /kg of DM consumed that can be useful in fermentation by microorganisms in the rumen. A feed or ration with positive UFP value is one that has more fermentable energy present than that needed for transforming the ammonia into microbial proteins. While a feed with a negative UFP value is that which has less fermentable energy to transform into microbial proteins eg. Maize has UFP of 11.8 which means that 11.8 gms of urea is useful for fermentation by rumen bacteria for 1 kg of maize. The efficiency of urea utilization depends on the composition of the ration and the feed management Composition of ration 1. Urea can be utilized efficiently with high TDN (75%) when the CP is below 1213%.With low energy feeds (TDN <60%) the urea can be utilized if the CP is below 7%. This is so because urea can be utilized only if a carbon skeleton is available for amination. 2. Minerals of the feed affect the utilization of urea since many of them are constituents of key co-factors involved in the production of microbial protein. Additionally sulphur must be provided for the formation of sulphur amino acids. 3. Feed additives have little effect on urea utilization. 4. Low level of antibiotics if added to feed does not reduce urea utilization. Managerial factors The animal to be fed urea must be given an initial period of adaptation because: 1. The microbial population of the rumen change slowly 2. The rate of urea hydrolysis is less 3. Mixing of urea is very important because if the urea is not properly and uniformly mixed it can result in urea toxicity. 4. Frequency of feeding can affect the efficiency of urea utilization. Urea should not be fed if it is used in the diet infrequently. Methods of feeding urea Urea can be provided in the feed taking into consideration the following factors 1. Protein need of the animal 2. availability and cost of urea 3. availability of energy and amount of plant protein used in the ration 4. cost of processing and mixing Urea mixed in concentrates: It can be incorporated in the concentrate portion of the ration of the growing, lactating and beef cattle. It is incorporated at 1% of the total DM or 1% in concentrate mixture. The maximum safe limit is 136 g urea/animal over 260kg BW.

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Liquid supplements: Urea is mixed uniformly in liquid molasses along with minerals and vitamins. Mix 2.5 parts urea with 2.5 parts water and add vitamin AD3 @ 25g/100 kg liquid feed. Add common salt 1 part and mineral mixture 2 parts to 92 parts molasses. Give an adaptation period of 15 days before starting on liquid supplement. Animal should be fed 1 kg of dry forage before giving liquid feed ad lib. Fresh drinking water should be freely available. Urea mixed with silage-If whole maize plant is ensiled at 35-40%DM then urea can be added @0.5% of wet weight. This increases the CP of the silage by 5 points. Urea added to dry roughages- a solution of 10 kg molasses and 2 kg urea in 10 kg water is sprayed on 100 kg straw. The treated straw can form a maintenance ration when supplied with 2% mineral mixture, 1% salt an vitamin AD3. About 8 kg of this straw/animal/day can supply enough nitrogen to meet the protein requirements for maintenance. Urea in salt block: This is another simple method of providing urea. Several combinations of ingredients can be used to make the block by the hot or cold process. Slow release urea products: several products in which urea is given heat treatment along with sugarcane molasses this helps in reducing the possibility of toxicity of urea and decreasing the release of ammonia thus reducing the possibility of urea toxicity. Urea can be mixed with brans when hot and be used as replacement of cakes. Guidelines for using urea in feeds 1. Urea is best utilized in a well balanced high energy ration. It is not well utilized with low quality roughages. This is so because the carbohydrate in grasses and hays is slowly available and the microbes cannot use the urea efficiently. The ration should also be balanced in vitamins and minerals. 2. Factors essential for optimum urea utilization 1. Mix urea thoroughly 2. Feed urea based rations to mature animals and not to any monogastrics. 3. Provide ready source of energy such as molasses or grains 4. Supply adequate and balanced minerals 5. The nitrogen to sulphur ratio should be 15:1. 6. Incorporate Lucerne meal to provide unidentified factors to stimulate microbial protein synthesis. 7. Include salt for palatability @0.5% of the complete ration and 3.5% I the protein supplement to increase the palatability. 8. Provide vitamin A along with other vitamins. 9. Accustom the animal to urea feeding gradually and feed urea at equal intervals for regular uniform supply of urea. . The adaptation period is minimum 5-7 days.

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10. Do not over feed urea. Generally urea is given @ 1/3 of the total nitrogen in the ration. 11. Do not feed urea to high yielders because at this level of production the microbes are unable to synthesize enough protein to meet the needs 12. The best utilization of urea can be made with cereal straws and stovers as compared to leguminous crops. 13. Dont use urea as a sole source of nitrogen for young rapidly growing animals and high producing lactating animals. 14. Never use urease containing seeds in the ration of urea containing ration eg. soybean or raw beans will liberate ammonia with urea which can be toxic to the animals. Toxicity: When urea is fed in excess large amount of ammonia is liberated in the rumen. The pH of the ruminal fluid increases and this facilitates the passage of ammonia across the rumen wall. If the ammonia absorbed is greater than the capacity of the liver to convert the ammonia to urea the ammonia accumulates in the blood and if it is more than 1 mg/100 ml in cattle toxicity occurs. Symptoms of ammonia toxicity are tetany, respiratory difficulty, bloat, excessive salivation, ataxia, convulsions and bellowing. If the animal is not treated it can die within 30 min. The common treatment is to drench 20-40 liters of cold water which inhibit ureolytic activity of the rumen. Or the animal can be drenched with 4 liters of dilute acetic acids like vinegar along with cold water. Animals become susceptible to urea toxicity due to Poor mixing, error in ration formulation, inadequate adaptation period, less water, feeding urea with low quality roughages, rations raising rumen pH. Biuret: This compound is produced by heating urea to form a crystalline compound. It contains 30% nitrogen equivalent to 188% CP. The compound is expensive and therefore not used. The advantage of this product is that it does not cause urea toxicity even at higher level.

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Anti-nutritional factors These are secondary metabolites of plants. They impair the physiological functions of the animals after ingestion. Toxicity or poisoning occurs when the level is higher than the detoxification level. They can reduce nutritional quality of feed by limiting the nutrient availability. Protease inhibitors- Legumes have trypsin and chymotrypsin enzyme inhibitory effect. Symptoms- reduced growth, pancreatic hyperplasia and lower production. The inhibitor is heat labile. Tannins- They are polymeric phenols and may be hydrolysable or condensed. Hydrolysable tannins are toxic. They are present in mustard cake, mango seed kernel, sorghum fodder, babul pods, salseed cake, tamarind seeds etc. They depress VDMI due to bitter and astringent taste, digestion of protein, hemi-cellulose, damage liver, kidney and bone marrow. Remedy-Washing with water, soaking, boiling cooking or soaking in salt solution (0.5N NaHCO3, Na2CO3, Na2SO4 and Na2S), organic solvents (acetone, methanol, formaldehyde). Phytic acid. Phytates are salts of phytic acid found in soybean, cotton seed, wheat, wheat bran etc. It binds with Zn, Fe, Mn. Calcium enhances the formation of Znphytate complex in the duodenum. About or more of P in cereals grains is as phytin. The availability of phytin-P depends on vitamin D, Ca:P ratio, Ca and Zn in feed etc. The P in inorganic form are available upto 80%. In ruminants enzyme phytase is produced by microbes. Phyto-estrogens- They are isoflavonoid or coumestan derivatives that disturb the sexual cycle of females. They are found in red clover, Lucerne, white clover, berseem, trifolium etc. Phosphate and Zn deficiency in soil can increase the isoflavone concentration in plants. Saponins- They are biologically active glycosides of steroidal or triterpenoidal nature It is found in Lucerne, white and red clover, soybean, peas, berseem etc. Saponins on hydrolysis yield sapogenins. Effect- hemolysis of RBC, bitter taste, bloat, reduces feed intake and palatability. Remedy is adding bhusa or spraying oil on the fodder. Cyanogenetic glycosides/ HCN/cyanogens- The precursors of cyanogenetic glycosides are the amino acids and their precursors. Cynogens are hydrolysed to prussic acid by plant enzymes. The forages high in HCN are sorghum before flowering, Sudan grass, sweet potato, chick pea, linseed cake, mustard cake and sugarcane tops. The content is higher when the plant grows rapidly after retarded growth, trampled frost bitten, wilted or after a hailstorm. High soil N and low P and herbicides 2,4-D are conducive for higher HCN content. It affects certain nervous system and cytochrome oxidase system. Symptoms are nervousness, trembling, abnormal breathing, blue coloration of mouth lining, convulsions, and respiratory failure. Injection of NaNO3 or NaHSO4 are given to cure. Ruminants are more prone to HCN poisoning as in the monogastric the enzyme is denatured with HCl in the stomach. Remedy-Hay making and washing with water.

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Alkaloids- They are physiologically active basic compounds found in canary grass, guar and their products, legume seeds. They are bitter and cause diarrhea, jaundice, muscle weakness etc. Boiling in water reduces the ill effects. Toxic amino acids- They are neurotoxin. Seleno-amino acids cause alkali disease and blind staggers. Mimosine- It is a toxic free amino acid called leucinine found in subabul (3-5%) and other legumes. It causes reduced growth, excessive salivation, gum erosion, lesion of tongue, goiter, loss of fertility etc. The harmful effect can be reduced by supplementation with FeSO4, Cu and Zn. Hot water dipping or soaking for 24 hrs can reduce the mimosine. Oxalates- Oxalates are found as soluble salts of Na and K or insoluble salts of Ca. Insoluble salts are toxic and crystallize in kidney and rumen wall. It is found in N-B hybrid, sugar beet leaves, buckwheat, paddy straw, water hyacinth, spinach, lettuce, agro industrial by-products. It is corrosive and a systemic poison. Young plants have more than mature plants. Effect-negative Ca balance, impaired P, Mg level. Ruminants are less prone. Symptoms are rapid and labored breathing, depression , weakness, coma, death, convulsion and tetany. Remedy is wilting, ensiling, spray with 0.5% NaOH, water washing, delayed harvest, feeding with legume fodder. Nitrate and nitrite toxicity- It is caused by indiscriminate use of N-fertilizer, irrigation with effluent water containing high N, cattle shed washings or sewage water, Mo and S deficiency, 2, 4-D application, lack of sunshine (reduction of nitrate to nitrite is controlled by sunlight). Nitrate on ingestion is converted to nitrite and enter blood stream to oxidize Fe+2 iron of Hb to Fe+3 (mathemoglobin), which cannot transport O2 in the blood. It also affects Vitamin A supply, growth and production. Symptomstrembling, salivation, teeth grinding, uneasiness, sever cyanosis, sometimes chronic hypocalcaemia, convulsions, paralysis etc. It may lead to death and abortion. High nitrate can be found in oat hay, green oats, barley, wheat, rye hay, rye grass, turnip tops, rape, sugar beet tops, linseed plant, N-B hybrid, maize, spinach, lettuce, beet root etc. During drought NO3 accumulates in soil and is absorbed on irrigation. Stem contain more nitrates than leaves and flowers. Nitrite is more lethal than nitrate. The rate of ingestion affects the toxicity, if fast then more toxicity. Remedy-Delay cutting of dark green luxurious growth of leaves, ensiling maize with 0.25% FeSO4 or 1% legume. Goitergens- All brassica like kale, cabbage, rape, soybean, peas, ground nut, rapeseed and mustard cake have glucosinolates/goitergens. It reduces I incorporation in the hormone thyroxine as well as its secretion. Hydrolysis of glucosinolates causes this effect. Hydrolysis is less in ruminants. Effect-Low birth weight and abortion, remedy is supplementation of iodine. Urease- Water melon and soybean meal has this activity. Gossypol- It is a yellow polyphenolic pigment found in the pigment glands of cotton seed. Free gossypol is physiologically toxic but not the bound gossypol. Effectreduced growth, feed intake, cardiac irregularity, reduced oxygen carrying capacity of blood and affects liver enzymes. It reduces lysine availability due to the reaction of

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aldehyde group of gossypol with NH3 group of lysine. Heating or boiling is recommended. Resin- Castor leaves particularly the mature leaves have resin which contains hemagglutalinins that agglutinates RBC. It is inactivated by moist heat. Anti vitamin A- soyabean enzyme lipoxygenase-oxidation of carotene Anti vitamin D in soybean Anti vitamin K This occurs on feeding moldy hay or silage and called sweet clover disease. It causes haemorrhagic condition in cattle due to dicumarol which reduces prothrombin level in blood and interfere in blood clotting. Photo dynamic agents- Feeding of kale, Lucerne hay, rape fodder damaged by water or fungus can cause photosensitization in livestock which results in redness on skin or ear, eyelids, muscle of face and teats. Mycotoxins- These are toxic fungal metabolites produced during any stage of production, processing, transportation and storage. It is found in groundnut cake, maize, jowar, fishmeal and cotton seed cake. High moisture(>14%) and poor storage (poor ventilation, temperature, high humidity) encourage production. Sun drying, organic acids (propionic, formic), feed acidifiers, addition of zeolite (hydrated Na-Caalumino silicate) control it. Aflatoxin- Produced by filamentous fungi Aspergillus flavus. The toxins include B1, B2, G1, G2 and M1. It causes liver cell carcinoma. Aflatoxin B1 is most potent causing liver cancer, bile duct proliferation, liver tumor, liver damage. Tolerance limit and effect of mycotoxin Mycotoxin Tolerance limit mg/kg Effect BW Aflatoxin 0.25 Growth depression, immuno suppression, subcutaneous and intra muscular haemorrhage OchratoxinsA 0.20 Growth depression, immuno suppression, impared mineralization, vitamin deficiency symptoms and kidney and liver damage T2 toxin 0.50 Growth depression Vomitoxin 1.00 Subcutaneous and intra muscular haemorrhage, diarrhea and bone changes Toxicity due to minerals Heavy metals pollution may be either by absorption from the soil (effluent irrigation Pb and Cr_ or adsorbed o leaf surface through pollution (Pb and F). Milk and liver, kidney and heart accumulate the heavy metals. Selenium- Seleium at 0.5-03.0 ppm in the diet of animal is toxic. Toxicity symptoms are stiffness of joints, hair loss and deformity of hoof. Molybdenum-The Mo is from steel and aluminium industry. Toxic symptoms are diarrhea, reproductive disorder and deformity of bone.

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Lead- comes from highways, smelter factories and tannery effluent irrigation. Fluorine- Found in ground water and rock phosphate. The toxic dose is 20 mg/kg dry feed.

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