DEPARTMENT OF LIVESTOCK PRODUCTION ADVANCED ANIMAL NUTRITION LP 3201

B.K.H.F.A. WICKRAMASURIYA 07 / AG / 027 EG 481 SABARAGAMUWA UNIVERSITY OF SRI LANKA

TABLE OF CONTENTS
1 Practical No: 02 .................................................................................................................. 3 1.1 Feedstuff Evaluation ................................................................................................... 3 Physical evaluation .............................................................................................. 3 Chemical evaluation............................................................................................. 4 Biological evaluation ......................................................................................... 10

1.1.1 1.1.2 1.1.3 2

Practical No: 03 ................................................................................................................ 12 2.1 Feed processing/upgrading techniques...................................................................... 12

Practical No: 04 ................................................................................................................ 16 3.1 3.2 Preparation of molasses urea blocks ......................................................................... 16 Processing of feed ingredients and manufacture of feed rations ............................... 17

PRACTICAL NO: 02

1.1 Feedstuff Evaluation 1.1.1 Physical evaluation

Objective: Analyze for physical characters o paddy straw, grass hay, silage, common feed ingredients and concentrate such as poultry, pigs and dairy ration. Method: For physical evaluation we have used upgraded paddy straw which is treated by alkaline solutions of urea and NaOH with three types of concentrations. We have prepared seven treatments with three replicates and after 24 hours, have checked its following characters. Appearance Smell Texture Palatability Inert materials such as weeds, dust, moulds, insects etc. Replicates A1 A2 A3 B1 B2 B3 C1 C2 C3 D1 D2 D3 E1 E2 E3 F1 F2 F3 color smell Yellowish NH3 odor green to brown Appearance texture Soft, can separate for fiber Soft, can separate Can separate for fiber Soft can be separated from fiber Soft, can be separate moisture pH > 72 % 6.8

Treatments Paddy straw only Paddy straw + 2% urea Paddy straw + 4% urea Paddy straw + 6% urea Paddy straw +2% NaOH Paddy straw +4% NaOH

Yellowish Strong NH3 & green butyric acid color odor Yellowish Strong NH3 green and light butyric odor yellowish green to green Yellowish green to brown Slight butyric & strong ammonia Slight butyric & strong ammonia

> 72 %

> 72 %

8.5

> 72 %

10.55

> 72 %

9.44

Yellowish Slight butyric green to acid smell brown

Can seperate

>65 %

10.5

1.1.2 Chemical evaluation

Performed analysis for following components of given feed and feed ingredients. 1. Dry Matter content 2. Ash content 3. Crude Protein content 4. Crude fat or Ether extract content 5. Gross energy content 6. Moisture content

1.1.2.1 Determination of Dry Matter

We used direct Oven drying method. Procedure Weigh the empty dry crucible. Then weigh fresh forage sample with crucible. Forage sample with crucible were placed in the oven. Dry for 4 hours at 100+1 degree Celsius. Crucibles were removed & place in a dedicator until cool. Then crucibles were weighed with dried forage residue. Blank test was carried out same time.

Calculation [(weight of dish + ash) - (weight of dish)] - (mean blank) -------------------------------------------------------------------[(weght of dish + forage) - (weight of dish)]

Ash % =

x 100

Dry matter % (rice bran) = [(20.3596 19.3505) / (20.3903 19.3505)] X 100% = 97.048%

Dry matter % (soya bean meal) = [(42.0049 = 92.61%

40.1363) / (42.154 40.1363)] X100%

Dry matter % (limestone)

= (43.6211 39.86) / (43.642 39.86) X100% = 99.45%

Dry matter % (maize)

= (17.1467 15.5871) / (17.2818 15.5871) X100% = 92.03%

Conclusion limestone dry matter content is higher than all other ingredients and calculation of it much essential for the food processing.

1.1.2.2 Determination of Ash

Procedure Weigh the empty dry crucible. Then weigh fresh forage sample with crucible. Crucibles with forage sample were placed in the muffle Furnace. Then it leaved for 12 hours at 5000C. Then crucibles were removed & place in a dedicator until cool. Crucibles with ash residue were weighed. Blank test was carried out in same time.

Calculation Ash % = [(weight of dish + ash) - (weight of dish)] - (mean blank) -------------------------------------------------------------------[(weght of dish + forage) - (weight of dish)] x 100

1.1.2.3 Determination of Nitrogen (Total) and Estimation of Protein in forage by Kjeldahl Method
Objective:-To detect protein content in feed samples Procedure:-We got 4 feed samples and a control. They are Broiler finisher Calf feed Layer feed Swine grower feed 0.5 g 0.5 g 0.5 g 1g

After measuring, sample was digested with sulphuric acid and catalyst mixture. Digested solution was distilled and NH3 was liberated in boric acid. Sample was titrated with HCl acid. Titration Titrate, H3BO3 receiving solution (distillate) with standard acid (0.2 M HCl) solution to first trace of pink (end point is permanent grey purple color)

Feed type Broiler feed Layer feed Calf feed Control

Titer volume 0.2ml 2.3ml 4.5ml 0.2ml

Observation:Sample was become very dark after digestion. When do titration sample became from red color to pink to grey purple color. Calculations: Total N (g/100g) = 14.01 * [Sample titre Blank titer] * Sample weight * 10 Titrate molarity

Layer feed Total N = = [14.1 X (2.3 -0.2) X 0.2] -----------------------------0.5 X 10 1.1844 X 6.25 = 1.1844 = 7.4025 %

The crude protein content

Calf feed Total N = = [14.1 X (4.5 0.2) X 0.2] --------------------------------- = 0.5 X 10 2.4252 X 6.25 = 2.4252 15.1575 %

The crude protein content

Discussion: We have to remove kjeldahl flask with swine grower feed because it is over boiled at digestion. Sulfuric acid, Sodium hydroxide, and hydrochloric acid can cause severe burns. Catalyst powder can cause irritation. So, we should handle them carefully. Should take special care when digestion and distillation. When place the flask under the condenser of the distillation unit so that the end of the condenser tip should be well below the boric acid solution surface. Otherwise the NH3 gas can escape. Normally in layer feed, 15-16% protein available but after checking protein content of our layer feed sample we got 7.4% protein. This can be because of some errors when measuring feed sample and some errors when doing titrations.

1.1.2.4 Determination of crude fat

Objective: to measure the fat content of feed samples Method: Weighed dried feed samples and put into an extraction thimble. Then place the thimble inside the Soxhlet apparatus After weighing solvent flask (105.015g), add the 140ml of ether into that flask and connected it beneath the apparatus. Adjust the heating rate and after 6 hours remove the thimble and reclaim ether using the apparatus. Complete the removal of ether on a boiling bath and dry flask at 1050C for 30 minutes. Finally cool in desiccators and weigh.

Calculation Weight of coconut poonac Weight of the solvent flask with feed Kept it in apparatus 6 hours to condensation Weight of flask with extract fat Weight of fat =105.2183g =105.2183g - 105.0163g =0.202g = 2g =105.0163g

Crude fat (% of DM) =

Weight of fat

X 100

Weight of sample

=0.202*100/2 =10.1%

1.1.2.5 Determination of gross energy content from the Adiabatic Bomb Calorimeter (bombing)
Objective: Determination of energy content of the feed. Procedure: We measured the feed samples and they are, Swine grower feed 1g Layer feed 1g Coconut poonac 1g Then grind solid materials to a fine powder consistency and make a pellet. Keep the pellet in the metal cup Measure the length of fuse wire and mount it on the cup-supporting frame and pour a few ml of distilled water into the bomb. Place the bomb into the supporting frame and insert the cup with feed into the bomb. Connect the oxygen cylinder to the bomb. First open cylinder main valve, then the connection valve and allow pressure in the bomb to build up to 25ATM.

 Pour 2.00 Kg of distilled water, with a temperature of between 24 and 250C into the container. Place the bomb in the container of water and switch the calorimeter to run and allow equilibration of temperature. Press ignition switch and record water temperature in container before ignition. Record final temperature after the temperature stops changing. Open the calorimeter and remove the bomb from the bucket with the special tool and before removing the cap release pressure valve slowly. Then remove the feed cup from bomb and measure reminder of fuse wire left on igniter.

Calculations: Layer feed Initial length of platinum wire Remained length after burning Amount of waste To burn 1cm of wire need 2.3cal of energy Amount of energy need to burn 5.1cm Energy production with 2L of water 5.1 * 2.3 1.8 * 2 = 11.73 cal = 1.80C = 3.6 kcal = 3600cal Energy content of swine grower feed 3600 11.73 = 3588.27 cal Temperature difference of bomb calorimeter 26.8 25 7 cm 1.9 cm 7- 1.9 = 5.1cm

Coconut poonac Initial length of platinum wire Remained length after burning Amount of waste To burn 1cm of wire need 2.3cal of energy Amount of energy need to burn 8cm Energy production with 2L of water Energy content of swine grower feed 6 * 2.3 1.7 * 2 3400 13.8 = 13.8 cal = 1.70C = 3.4 kcal = 3400cal = 3386.2cal Temperature difference of bomb calorimeter 28.5 26.8 7 cm 1cm 7- 1 = 6cm

Swine grower feed

Initial length of platinum wire Remained length after burning Amount of waste To burn 1cm of wire need 2.3cal of energy Amount of energy need to burn 5cm Energy production with 2L of water

7 cm 2cm 7- 2 5 * 2.3 1.6 * 2 = 5cm = 11.5 cal = 1.60C = 3.2 kcal = 3200cal

Temperature difference of bomb calorimeter 30 28.4

Energy content of swine grower feed

3200 11.5

= 3188.5cal

1.1.3 Biological evaluation

1.1.3.1 Two stage Tilley & Terry method
Objective: measure the microbial digestion of feed stuffs. Apparatus and materials: Feed sample, rumen fluid, Buffer solution A, Buffer solution B, incubation tubes, shaking water bath, sintered glass crucibles, desiccators, weighing scale, filtration unit, oven Preparation of buffer solution Buffer A & B is used for maintenance of pH at 6.8. Buffer Solution A Compound g/liter

KH2PO4 10.0 MgSO4 7H2O 0.50 NaCl 0.50 CaCl2 2H2O 0.10 Urea (reagent grade) 0.50 Dissolve in liter of distilled water and should store at 39 C Buffer Solution B Compound g/liter

Na2CO3 15.0 Na2S 9H2O 1.0 Dissolve in liter of distilled water and should store at 39 C Procedure

Add 0.5g feed into incubation tube. Then add 65ml of incubation medium (Buffer & rumen fluid in ratio of 4:1). Include a blank tube with incubation medium. Close each tube with a rubber stopper with a releasing valve. Incubate for 48 h in a shaking water bath at 39 C. Then, add acidified pepsin solution (0.12g pepsin in 5ml of 3.3 M HCl for each tube). Incubate for another 48h. Filter the residue under vacuum using sintered glass crucibles. Dry the crucible with residue for 2hrs at 100C.

Calculation Digestibility % = 100 - (Residual weight) Sample weight Treatment A B1 B2 B3 C1 C2 C3 D1 D2 D3 E1 E2 E3 F1 F2 F3 G1 G2 G3 Residual weight 0.5451 0.3657 0.4634 0.6993 0.7762 0.7963 0.8172 0.714 0.9574 0.7404 0.7361 0.6484 0.3157 0.6434 0.7733 0.7941 0.1713 0.0861 0.1833 Sample weight 1 1 1 1 1.347 0.982 0.9923 1 1 1 1 1 1 1 1 1 1 1 1 Digestibility% 45.49 63.43 53.66 30.07 42.37 18.91 17.64 28.6 4.26 25.96 26.39 35.16 68.43 35.66 22.67 20.59 82.87 91.39 81.67 x 100

Observation

After incubation reduced sample weight. These reductions differ with different samples. It happens due to alkaline treatment. Best digestibility in samples G. This sample was treated with 6% NaOH. Discussion In this method we provide artificial rumen conditions for feed digestion. Need to put samples to incubation tube immediately. If not enter O2 to the sample and inactivate microbial activity. Grind feed sample to facilitate easily digest. Treat paddy straw by urea, NaOH like alkali solutions digestibility increase due to loosening of cellulose, hemicelluloses, lignin, suberin like materials.

PRACTICAL NO: 03

2.1 Feed processing/upgrading techniques

2.1.1 Effect of alkaline treatment on Paddy straw upgrading
Materials: polythene bags, rubber bands, paddy straw, chopping knives, chopping boards, mixing plastic trays, urea, NaOH Procedure; 1g grinded feed sample was added in to incubation tube. 65ml of incubation medium was put to incubation tube. Blank tube was included with incubation medium Each tube was closed with regiform stopper with a releasing valve. Each incubation tube were Covered with aluminum foil. Each sample was incubated for 48 hours in a shaking water bath at 39oC After incubation, Added acidified pepsin solution (0.12g pepsin in 5ml of 3.3 HCl for each tube) Incubated for another 48 hours Filtered the residue under vacuum using sintered glass crucibles Dried the crucible with residue (2 hours / 100 oC)

Group A B

Treatments 50g Paddy straw only 50g Paddy straw + 2% urea

Replicates 3 3

C D E F G

50g Paddy straw + 4% urea 50g Paddy straw + 6% urea 50g Paddy straw + 2% NaOH 50g Paddy straw + 4% NaOH 50g Paddy straw + 6% NaOH

3 3 3 3 3

Wrap properly & tie up with rubber bands. Analyse each treatment for crude protein, neutral detergent fiber and acid detergent fiber contents at 24 and 48 hour periods. Preparations of alkali solutions are done by dissolve each level of urea and NaOH in 100 ml of water.

Observations
Character Color Smell A1 Yellowish green color Strong ammonia & slight butyric acid smell A2 Yellowish green color Strong ammonia & slight butyric acid smell A3 Brown green Strong ammonia & slight butyric acid smell

Character Color

C1 Yellowish green color

C2 Yellowish green color

C3 Brown green

Texture Moisture PH(by litmus) PH(by PH meter)

Soft material. Can be separated from fiber >72% 9 8.9

Soft material. Can be separated from fiber >72% 9 8.94

Soft material. Can be separated from fiber ~65% 8 8.72

Smell

Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9 8.9

Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9 8.94

Texture Moisture PH(by litmus) PH(by PH meter)

Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber ~65% 8 8.72

Character Color Smell

B1 Yellowish green color Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9

B2 Yellowish green color Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9

Texture Moisture PH(by litmus)

B3 Brown green Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber ~65% 8

Character Color Smell

D1 Yellowish green color Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9 8.9

Texture

D2 Yellowish green color Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9 8.94

D3 Brown green Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber ~65% 8 8.72

Moisture PH(by litmus) PH(by PH meter)

Character Color Smell

Texture Moisture PH(by litmus) PH(by PH meter)

E1 Yellowish green color Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9 8.9

E2 Yellowish green color Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber >72% 9 8.94

E3 Brown green Strong ammonia & slight butyric acid smell Soft material. Can be separated from fiber ~65% 8 8.72

Character Color Smell Texture

F1 Yellowish brown Slight butyric and ammonia smell Soft material. Can be separated from fiber Tend to be 65% 10.57

F2 Yellowish brown Slight butyric and ammonia smell Soft material. Can be separated from fiber Tend to be 65% 10.5

F3 Yellowish brown Slight butyric and ammonia smell Soft material. Can be separated from fiber Tend to be 65% 10.52

Moisture PH(by PH meter)

Character Color Smell Texture Moisture PH(by PH meter)

G1 Brown to black Good acid smell Soft material. Can be separated from fiber >72% 10.29

G2 Brown to black Good acid smell Soft material. Can be separated from fiber >72% 10.41

G3 Yellowish brown Good acid smell Soft material. Can be separated from fiber ~65% 10.52

Discussion;

When doing this practical we should supply condition like inside the rumen to achieve accurate results. Sample should be grinded in to fine particle like animal chewed. Otherwise digestibility will not be happened well. During incubation its anaerobic conditions should be provided well. But in our practical it is not supply well. So anaerobic digestibility is not happen well. Due to entering oxygen in to digestive tube anaerobic microbes can die. After treating paddy straw with urea, NaOH like alkali solutions digestibility should be increased than normal due to this reasons as the alkali solutions help to loosen binding in cell wall in cellulose, hemicelluloses, lignin, suberin like materials. Then we can say it was happened in high percentage of NaOH treatment. Others are not significant variation than control, it may be due to practical errors.

PRACTICAL NO: 04

3.1 Preparation of molasses urea blocks

Procedure We make 1 kg molasses urea block. According to the above percentage we calculate the ingredients and mix. Measure the minerals Grind urea and common salts in to fine particles. Mix urea and salt in 40ml water and then mix with measured cement,CaO,MgO,

 Added the rice bran and molasses and mixed thoroughly . Finally fill in to suitable blocks and leave till develop the required hardness Calculations Rice bran Molasses Urea Common salt = 25*1000 g 100 = 50*1000 g 100 = 10*1000 g 100 = 5*1000 g 100 = 250 g = 500 g = 100 g = 50 g

MgO CaO Cement Water Observation

= 10*1000 g 100 = 4000 ml*100 g 10000 g

= 100 g

= 40 ml

Color changed to black due to adding molasses. Within 5-6 days it becomes harder due to cement. So after 5-6 days it can be used for animals. Discussion At mixing add less water amount. If not mixture liquefied and get longer time period for harder. We used blender to blend urea and common salts.

3.2

Processing of feed ingredients and manufacture of feed rations

Procedure Firstly should create a suitable formula Then gather all required ingredients Clean & dry the mixing floor Weigh the ingredients separately & accurately Mix micro ingredients in a covered container Put all other ingredients on the mixing floor Mix thoroughly with a shovel

Divide major ingredient mix into 2 halves Divide one half again into 2 halves Spread one quarter & mix with micro mixture Spread this mixture, add 2nd quarter and mix well Again spread this mixture, add remaining half and mix well Heap the final mixture and pack into bags or containers Ingredients % Rice polish Maize Soybean meal Meat & bone meal Limestone/Shell grit Dicalciumphosphate Lysine HCl DL Methionine Salt Coccidiostat Starter/Layer premix Total Energy (ME) Kcal/kg Crude protein % Crude fibre % Layer chick starters 30 40 15.2 3.5 10 0.5 0.1 0.15 0.25 0.05 0.25 100 2896 16.1 4.7

Calculations Rice polish Maize = 25 kg*30 100 = 25 kg*40 100 = 7.5 kg = 10 kg

Soybean meal

= 25 kg*15.2

= 3.8 kg

100 Meat & bone meal = 25 kg*3.5 100 = 25 kg*0.5 100 = 25 kg*0.1 100 = 25 kg*0.15 100 = 25 kg*0.25 100 = 25 kg*0.05 100 = 25 kg*0.25 100 = 0.875 kg

Limestone/shell grit Lysine HCl DL Methionine Salt Coccidiostat Starter/layer premix Observation;

= 0.125 kg = 0.025 kg = 0.0375 kg = 0.0625 kg = 0.0125 kg = 0.0625 kg

We prepared layer ration as a mass. The ingredients particles like maize and soya bean meal which we are used is little bit large. So it may create difficulties in eating for layers. The qualities of the some ingredients are also less. Discussion; In the mixing of the ration it should be done carefully to provide balanced ration for animals.