It is with great pleasure that I introduce this book on animal nutrition—an indispensable tool designed to guide students, researchers, and practitioners through hands-on exercises essential for a comprehensive understanding of this dynamic field. The study of animal nutrition is an evolving field that plays a crucial role in ensuring the optimal performance, productivity, and health of various species. This book is crafted with the intention of providing a comprehensive and accessible resource for students, researchers, and practitioners in the realm of animal science. Each chapter within this manual is crafted to provide step-by-step guidance on a range of practical exercises. Whether you are a novice seeking an introduction to the fundamentals or an experienced professional looking for the latest advancements, this book aims to cater to a diverse audience. The goal is to empower readers with the knowledge and skills needed to make informed decisions in the realm of animal nutrition, whether in the context of livestock production or companion animal care.
Dr. Raman Malik, a distinguished scientist with a profound academic background, earned a Ph.D. from the National Dairy Research Institute (NDRI) in 1993. Embarking on a remarkable career, he joined the Agricultural Research Service (ARS) in the same year. From 1993 to 1998, Dr. Malik contributed significantly as a scientist at the Indian Veterinary Research Institute (IVRI). Later, as a senior scientist at NDRI, Karnal, upto 2005, and subsequently at the Central Sheep and Wool Research Institute (CSWRI), Avikanagar, until 2008, he demonstrated expertise and dedication. Serving as a principal scientist and officer-in-charge at the Central Institute for Research on Buffaloes (CIRB), Sub-campus, Nabha, upto 2015, he made substantial contributions to the field. Currently holding the position of Principal Scientist at NDRI, Karnal. Dr. Raman Malik boasts an impressive scholarly record with over 250 publications in both national and international journals. Additionally, Dr. Malik has authored three books and contributed chapters to numerous other publications, showcasing a wealth of knowledge and experience in the agricultural and veterinary sciences.
Dr. Harneet Kour, a native of Jammu, is a budding professional with a strong academic background in veterinary science and animal nutrition. She completed her B.V.Sc. and AH from SKUAST-J. Driven by a passion for advancing knowledge in animal nutrition, Harneet pursued her M.V.Sc. in Animal Nutrition from GADVASU, Ludhiana. Her research during her M.V.Sc. focused on Dietary Cation-Anion Difference (DCAD) for buffaloes, where she worked on standardized of DCAD level for effective nutritional management of buffaloes. She is currently pursuing her Doctor of Philosophy (Ph.D.) from the National Dairy Research Institute (NDRI) in Karnal. Her doctoral research is aimed to conduct rumen studies in a controlled laboratory environment. Her accomplishments include receiving accolades, such as the Best Thesis Award, Best Master’s Research Award and a gold medal for her outstanding academic performance during her post-graduate studies.
Dr. Parul Rana, born amidst the breathtaking landscapes of Himachal Pradesh is not only an arid researcher but also a dynamic and ambitions professional who stands at the beginning of a promising career. She achieved a stellar district topper position with a 10 CGPA in 10th CBSE examinations. Armed with a bachelor’s degree in veterinary science and animal husbandry (BVSc & AH) from Chaudhary Sharan Kumar Himachal Pradesh Agricultural University. She further learned expertise in Animal Nutrition through a master’s degree (MVSc) at Lala Lajpat Rai University of Veterinary & Animal Sciences (LUVAS), Hisar on the topic of growth performance of broilers herbal methionine supplementations. Currently, she is immersed in the world of academia pursuing PhD at NDRI, Karnal. Beyond academic landscape, she is a proficient writer, a persuasive debater and professional Kathak dancer with a Prabhakar degree in the classical dance.
1. Cattle and Poultry Feed Evaluation Laboratory: Set up and Establishment ................................................................ 1
2. Laboratory Safety.............................................................................. 7
3. Washing and Cleaning Laboratory Glassware ......................... 11
4. Preparation of Solutions ................................................................ 13
5. Sampling and Processing of Samples .......................................... 27
6. Proximate Analysis ......................................................................... 31
1. Determination of Dry Matter (DM)/Moisture .................. 32
2. Determination of Ash/Organic matter............................... 33
3. Determination of Ether Extract (EE)/fat/oil/Crude Fat................................................................................... 34
4. Crude Protein Estimation as per Kjeldhal Method ........... 37
5. Estimation of Crude Fiber (CF) ............................................. 44
7. Analysis of Fibre Fractions and Cell Contents .......................... 47
Determination of Cell Contents & Cell Wall Constituents ..... 48
Determination of Acid Detergent Fibre (ADF) ......................... 50
Acid Detergent Lignin (ADL)....................................................... 51
Determination of Permaganate Lignin, Cellulose,
Insoluble Ash and Silica ................................................................ 52
Determination of Modified Acid-Detergent Fibre (MADF).... 55
Estimation of Pectin ........................................................................ 56
Estimation of Total Dietary Fibre (TDF) ..................................... 57
Estimation of Non-Starch Polysaccharides ................................ 58
Determination of Starch ................................................................ 58
Determination of Free Sugars ....................................................... 60
Determination of Glucans Principle ............................................ 61
8. Protein and Carbohydrate Fraction As Per CNCPS Model ... 63
1. Estimation of Non-Protein Nitrogen (NPN) ....................... 64
2. Soluble Nitrogen and Protein ................................................ 66
3. Determination of Acid-Detergent Insoluble Nitrogen (ADIN)...................................................................... 67
4. Dtermination of Neutral Detergent Insoluble Nitrogen (NDIN)...................................................................... 68
9. Estimation of Gross Energy ........................................................... 73
(i) Types of Bomb Calorimeter ................................................... 73
(ii) Gross Energy (GE) of Feed and Faeces ................................ 75
(iii) Gross Energy of Urine ............................................................. 75
(iv) Gross Energy of Milk ............................................................... 76
(v) Ballistic Oxygen Bomb Calorimeter ..................................... 77
10. Rumen Fermentation Parameters ................................................ 81
1. Estimation of Total Volatile Fatty Acids (TVFA) .............. 82
2. Individual Volatile Fatty Acids by Gas Liquid Chromatography ..................................................................... 83
3. Estimation of Lactic Acid in Rumen Liquor....................... 85
4. Microscopic Counting of Protozoa in Rumen Liquor ...... 87
5. Bacteria Counting Technique in Rumen Liquor................ 88
6. Estimation of Rumen Fluid Volume (RFV) ......................... 90
7. Estimation of Bacterial Production Rate by Singh et al. (1984)..................................................................... 93
8. Kinetics of Digestion and Passage Rate .............................. 94
9. Rumen-evacuation Technique .............................................. 95
10. The Use of Markers to Estimate Passage Rates ................. 97
11. Enzyme Assay................................................................................ 103
(i) Cellulase Assay ...................................................................... 103
(ii) Protease .................................................................................... 105
(iii) Xylanase Assay ...................................................................... 108
(iv) Alpha Amylase Assay (Starch-Iodine Method) .............. 110
12. Physical Properties of Feed Ingredients .................................... 113
(i) Bulk Density ............................................................................ 113
(ii) Angle of Repose ..................................................................... 113
(iii) Coefficient of Friction of a Feed or Feed Ingredient ....... 115
(iv) Determination of Specific Density of a Feed or Feed Ingredients .............................................................................. 117
(v) Compaction of Feed Ingredients ......................................... 118
(vi) Determination of Hardness of Feed Ingredients ............. 119
(vii) Specific Weight & Durability of Densified Feed Products ......................................................................... 119
(viii)Specific Weight of Wafers .................................................... 119
(ix) Determination of Durability of Wafer ............................... 121
(x) Specific Weight of Pellets and Crumbles .......................... 122
(xi) Methods of Expressing Particle Size of Feed .................... 123
(xii) Determination of Particle Size, Number of Particles Surface Area Per Gram of Ground Sample ..... 125
(xiii)Surface Area & Number of Particles in Ground Feed .... 127
13. Feed Microscopy ........................................................................... 129
1. Sample Preparation............................................................... 129
2. Equipments ............................................................................. 130
3. Separation of Sample ............................................................ 131
14. Evaluation of Feed Mixing .......................................................... 135
(i) Evaluation of Feed Mixing by Salt ...................................... 135
(ii) Evaluation of Feed Mixing by Micro-nutrients ................ 137
(iii) Evaluation of Feed Mixing by Micro-tracer ..................... 137
15. Blood Analysis ............................................................................... 141
A. Collection & Preservation of Blood Samples .................... 141
1. Time of Blood Sample Collection .................................. 141
2. Sites of Collection ............................................................. 141
3. Commonly Used Blood Anticoagulants ...................... 141
4. Procedure of Blood Collection ....................................... 142
5. Preparation of Blood Smear ........................................... 142
6. Preparation of Blood Serum ........................................... 143
7. Preparation of Blood Plasma ......................................... 143
B. Haematology ........................................................................... 143
1. Total Erythrocyte Counts (RBCs) ................................. 143
2. Total Leucocyte Counts (WBCs) ................................... 144
3. Differntial Leucocyte Count .......................................... 145
4. Determination of Packed Cell Volume in Blood ........ 147
5. Determination of Erythrocyte Sedimentation Rate (ESR) .......................................................................... 148
6. Determination of Clotting Time of Blood .................... 149
7. Determination of Haemoglobin in Blood .................... 149
8. Blood Protein Estimation ................................................ 151
9. Estimation of Serum/Plasma Albumins and Globulins ............................................................................ 154
10. Bilirubin............................................................................. 155
11. Determination of Blood Glucose .................................. 156
12. Estimation of Glucose in Blood ..................................... 157
13. Determination of Serum/Plasma Urea ...................... 158
14. Estimation of Total Cholesterol in Serum/Plasma ... 160
15. Estimation of Creatinine in Serum/Plasma ............... 161
16. Ascorbic Acid .................................................................. 162
17. Pyruvic Acid .................................................................... 163
18. Calcium ............................................................................. 164
19. Phosphorus ....................................................................... 165
C. Blood Enzymes ....................................................................... 166
1. Calorimetric Method for Glutamic Oxaloacetic Trans-aminase (GOT) ....................................................... 166
2. Glumatic-Pyruvic Transaminase (GPT) Estimation .. 168
D. Methods for Determination of Ammonical Nitrogen in Blood .................................................................................... 169
E. Determination of Urease Activity in Blood ...................... 170
16. Milk Analysis ................................................................................. 173
Analysis of Major Milk Constituents Using Lactostar........... 173
Determination of Lactose in Milk by Polarimetric Method .. 175
Estimation of Urea in Milk .......................................................... 177
Estimation Milk Constituents ..................................................... 178
17. In vitro & in Sacco Studies ........................................................... 187
1. One-Stage Technique ............................................................ 187
2. Van Soest Method ................................................................. 191
18. Digestion Trial ............................................................................... 195
Digestion Trial Periods ................................................................. 195
Index Method of Determining Digestibility ............................. 198
Total Digestible Nutrients (TDN) ............................................... 199
19. Quick Analytical Feed Tests........................................................ 201
Test for Minerals in Finished Feeds ........................................... 201
Spot Test for Cobalt, Copper and Iron ..................................... 201
Toxic Constituents ........................................................................ 204
Cyanogenic Glycosides (HCN) in Feeds ................................... 204
Aflatoxin ......................................................................................... 204
Other Tests ...................................................................................... 205
Rock Phosphate or DCP .............................................................. 205
Carbonate ....................................................................................... 206
Meat Cum Bone Meal and Leather Meal ................................. 206
Salt (Sodium Chloride)................................................................. 207
20. Toxic Factors .................................................................................. 209
Estimation of Total Phenolics ..................................................... 209
Estimation of Saponins ................................................................ 210
Estimation of Gossypol................................................................. 211
Estimation of Mimosine ............................................................... 214
Estimation of Aflatoxin ................................................................ 216
Determination of Coumarins (A.O.A.C. 1970)....................... 219
Determination of Alkaloids ......................................................... 220
Determination of HCN ................................................................ 221
Estimation of Nitrates and Nitrites ............................................ 223
Estimation of Total Tannins ........................................................ 225
Estimation of Total Glucosinolates ............................................ 226
Determination of Aflatoxins by ELISA (ICRISAT method) . 227
21. Analysis of Minerals and Miscellaneous Tests ........................ 231
Determination of Calcium........................................................... 231
Determination of Phosphorus by Calorimetric Method........ 232
Estimation of Available Phosphorus ......................................... 233
Urease Test in Soybean Meal (Rapid Method)........................ 235
Urease Test in Soybean Meal ...................................................... 236
Urease Activity Procedure .......................................................... 236
Pesticides Test ................................................................................ 238
Rancidity Test (Rice Bran etc.) ................................................... 239
Decomposition Test (Eber’s Sulphide Test) .............................. 240
Appendices ..................................................................................... 241
References ....................................................................................... 247
