SOKOINE UNIVERSITY OF AGRICULTURE

COLLEGE OF AGRICULTURE

DEPARTMENT OF ANIMAL, AQUACULTURE AND RANGE SCIENCES

RESEARCH PROPOSAL ON
MILK CRUDE PROTEIN AND BUTTER FAT LEVEL OF FRIESIAN DAIRY CATTLE
AT MAGADU DAIRY FARM.

DEGREE PROGRAMME: BSc. ANIMAL SCIENCE

A SPECIAL PROJECT PROPOSAL
BY
MAHUNJA, NESTORY S
REGISTRATION NUMBER: ANS/D/2018/0140

SUPERVISOR NAME: PROF. V.R.M. MUHIKAMBELE

A research proposal submitted to the department of animal, aquaculture and range

sciences as a requirement for the degree of bachelor of science in animal science in animal
science at Sokoine University of Agriculture

JULY, 2021

1
 ABSTRACT
A study was carried out so as to obtain the average levels of milk components (specifically
Crude protein and Butter fat) for the Friesian cows found at magadu dairy farm at Sokoine
University of Agriculture. Milking was done by milking machine at Magadu dairy farm early in
the morning at 0600 hours and at evening 1800 hours. When there was no power, milking was
done by hand, by 3 experienced hand milkers. Cooper® milking salve, of Cooper
Pharmaceuticals (Kenya Ltd) was applied to the teats and adjacent areas of the udder after
milking to prevent cracking of the udder and teats. The amount of milk from each individual cow
was recorded. Milk samples were collected once a week (morning time) during test period.
Aliquot milk samples of 500 ml from each of the 12 animals were collected and mixed in a pre-
cooled container to make a composite sample totally of 6 liters which was then used for analysis
of several chemical components and physical properties at the laboratory. The milk samples were
analyzed for pH, milk density, milk composition with respect to BF, protein, TS and SNF. Milk
pH was determined by milk pH meter; milk density (lactometer test) was done by using
hydrometer (ILCA, 1988); BF was determined by using Gerber method (ILCA, 1988); N percent
was determined by Kjeldahl method outlined by Bremner and Mulveney (1992) and the CP
percent was calculated by the formula: CP% = N% x 6.38,

Then the average proportion for the three weeks was determined and the proportions was as
follows:

The average crude protein was 3.3517%, the average butter fat was 4.6%, the average total solid
(TS) was 13.7843%, the average solid not fat (SNF) was 9.1476, the average milk density was
29.63g/cc. so these data was recorded to the magadu dairy unit recording offices for future use.
The difference in milking day had no effect on the variations in the milk compositional levels.

i
 DECLARATION
I Mahunja, Nestory S. hereby declaring to the best of my knowledge, this report is my own work
and has not been submitted for any degree award at any higher learning institution.

……………………………. …………………………….

Mahunja, Nestory S Date

(Student)

The above declaration is confirmed by

……………………………. …………………………….

Prof. V.R.M. Muhikambele Date

(Supervisor)

ii
 COPYRIGHT
No part of this dissertation may be reproduced, stored in any retrieval system or transmitted in
any form or by any means; electronic, mechanical, photocopying, recording or otherwise without
a prior written permission of the author or Sokoine University of Agriculture in that behalf

iii
 ACKNOWLEDGEMENT
Firstly, I would like to thank the Almighty God for giving me strength, wisdom, grace, and
knowledge. My gratitude also goes to my supervisor Prof. V.R.M. Muhikambele from the
Department of Animal Science, Aquaculture and Range Sciences at Sokoine University of
Agriculture for his special guidance, encouragement, constructive comments and overall
supervision on my research project report.

I would like also to express my special appreciation to all lab technicians from DAARS for
conducting the chemical analysis of samples from this study.

Without forgetting the contribution from other supportive staff, my lovely family for their
prayers, economic support and spiritual encouragement in my study.

iv
 DEDICATIONS

This work is dedicated to my beloved family members Mr. Ashanti Azari, and Given
Mahunja for their support and encouragement.

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ABBREVIATIONS

BF Butter Fat
CP Crude Protein
TS Total Solids
SNF Solid Not Fat
SUA Sokoine University of Agriculture
FCM Fat Corrected Milk
ILCA International Livestock Centre for Africa
HDL High Density Lipoprotein
VLDL Very Low-Density Lipoproteins
LCFAs Long Chain Fatty Acids
DM Dry Matter
MCFAs Medium Chain Fatty Acids

vi
 TABLE OF CONTENTS
ABSTRACT................................................................................................................................................. I
DECLARATION......................................................................................................................................... II
COPYRIGHT............................................................................................................................................. III
ACKNOWLEDGEMENT.......................................................................................................................... IV
DEDICATIONS.......................................................................................................................................... V
ABBREVIATIONS.................................................................................................................................... VI

CHAPTER ONE.......................................................................................................................................... 1
1.0 INTRODUCTION................................................................................................................................. 1
1.1 BACKGROUND.............................................................................................................................................1
1.2 STATEMENT OF THE PROBLEM...............................................................................................................2
1.3 OBJECTIVES.................................................................................................................................................2
1.3.1 GENERAL OBJECTIVES...........................................................................................................................2
1.3.2 SPECIFIC OBJECTIVES.............................................................................................................................2

CHAPTER TWO......................................................................................................................................... 3
2.0 LITERATURE REVIEW...................................................................................................................... 3
2.1 MILK COMPOSITION...................................................................................................................................3
2.2 EFFECT OF BREED ON MILK COMPOSITION.........................................................................................3
2.3 DISEASES AND THEIR MEDICATION......................................................................................................4
2.4 COMPOSITION OF MILK FAT....................................................................................................................4
2.4.1 TRIGLYCERIDES.......................................................................................................................................4
2.4.1.1 BLOOD LIPIDS........................................................................................................................................5
2.4.1.2 DE NOVO FATTY ACID SYNTHESIS...................................................................................................5
2.4.1.3 CHOLESTEROL.......................................................................................................................................5

CHAPTER THREE..................................................................................................................................... 6
3.0 MATERIALS AND METHODS............................................................................................................ 6
3.1 DESCRIPTION OF THE STUDY AREA.......................................................................................................6
3.2 ANIMALS AND THEIR MANAGEMENTS.................................................................................................6
3.2.2 ANIMAL FEEDING....................................................................................................................................6
3.3 MILK COLLECTION AND ANALYSIS.......................................................................................................7
3.3.1. DETERMINATION OF TOTAL NITROGEN CONTENT IN MILK/MILK PRODUCT BY KJELDAHL
.............................................................................................................................................................................. 7
3.3.2. DETERMINATION OF FAT IN MILK....................................................................................................12

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3.4 STATICAL ANALYSIS……………………………………………………………………………………………………………….12
CHAPTER FOUR..................................................................................................................................... 14
4.0. RESULT AND DISCUSION............................................................................................................... 14

CHAPTER FIVE....................................................................................................................................... 16
5.0 CONCLUSION AND RECCOMENDATIONS...................................................................................16
5.1 CONCLUSION.............................................................................................................................................16
5.2 RECOMMENDATIONS...............................................................................................................................16

REFERENCES.......................................................................................................................................... 17

viii
 CHAPTER ONE

1.0 INTRODUCTION

1.1 Background
Milk is a nutrient rich liquid food that is produced by the mammary glands of most mammals.
It is important for the new born (Bascom, 2014).Common cattle milk components include
water, butter fat, protein, vitamins and minerals which are important for not only children but
also for the adult (Castle and Watkins, 2017).

Bovine milk composition is influenced by many factors including breed and genotype
(Gustavsson, Erison and Hanson, 2015; Coleman et al., 2016), nutrition (Walker, Howard
and Lind, 2016), milk yield (Palladino, Ashantey and Welbugey, 2017), cows/ heifer’s
physiological state (Gurmessa and Melaku, 2014), and stage of lactation (Stoop, Brown and
Tyrrel, 2015).

The two milk components (milk protein and milk fat (butter fat)) are very important
components regarding the nutritional quality of a milk. So, this research will be analyzing
those important components levels.

This research is of important as it will provide important information about the milk
composition so as the farm can justify the milk qualities of these Friesian cattle and if there
are any abnormalities in its compositional level some interventions may be done so as to
regulate the production into that of desirable so as to maintain desired nutritive quality of the
milk.

Milk crude protein level is likely to be affected by several factors such as the quantity of
energy level on the ration (Jelec, 2014; Emery, 2018), age of the animal, seasonal effects,
stage of lactation. Also, environmental and animal genetic play a role on the compositional
level.

Most of the factors affecting milk protein tend to affect the milk fat as well since these two
components are positively correlated. Also, the breed type tends to affect these levels
(O’Mahony, 2015; Morales, 2016).

These two milk components are very important now days in the market as they determine the
milk price at market (Murphy, Ashrat and Agustus, 2016; Bailey et al., 2018).

1
1.2 Statement of the Problem
At magadu dairy farm there is no current dagta on the average milk fat and milk protein
levels of the Friesian cattle, I have proven this by doing a simple survey where I asked the
farm manager of magadu dairy farm, Mr. Karoly and he told me that there is no current
analysis that have been done, thus they just have old and non-specific information regarding
the milk compositional levels of Friesian dairy cattle. so, this study will find the recent,
updated and specific data regarding average Friesian cattle milk components levels.

1.3 OBJECTIVES

1.3.1 General Objectives

 To obtain the current values of milk chemical composition of Friesian dairy cattle at
magadu dairy farm.

1.3.2 Specific Objectives

 To determine the average chemical composition, pH value and density of Friesian
dairy cattle at magadu dairy farm.
 To compare the values of various milk components with normal values and establish
the reasons for such differences.

2
 CHAPTER TWO

2.0 LITERATURE REVIEW

2.1 Milk Composition

In nature the role of milk is to nourish and provide immunological protection to the young
mammalian. It has been found that milk has been found to have many positive effects to
human health and it has been used as food for humans since prehistoric times (Bascom,
2014). The average gross composition of cow’s milk is as follows: 87.3% water (range of
85.5% - 88.7%), 3.9% milk fat or milk lipids or BF (range of 2.4 - 5.5%), 8.8% solids-not-fat
(SNF) (range of 7.9 - 10.0%), protein 3.25% (0.75% casein), lactose 4.6%, minerals 0.65% -
(Ca, P, citrate, Mg, K, Na, Zn, Cl, Fe, Cu) sulfate, bicarbonate, acids 0.18% - citrate, formate,
acetate, lactate, oxalate; enzymes - peroxidase, catalase, phosphatase, lipase; gases - oxygen,
nitrogen; vitamins - A, C, D, thiamine, and riboflavin (Watt and Merrill, 2014)

Table 1: milk components for different animal species

components Cow milk Goat milk Human milk
Protein 3.58 3.52 1.63
Fat 4.14 4.25 3.75
Total solids 13.19 13.00 12.57
Solid not fat 9.25 7.75 8.82
Lactose 4.96 4.27 6.98
Ash 0.71 0.86 0.21

2.2 Effect of Breed on Milk Composition

Milk composition varies with a number of non- nutritional factors including breed (Caroll et
al., 2006). Friesian, the breed with highest yield potentials has the lowest fat whereas the
lower yielding Jersey and Guernsey have the highest fat contents (O’Mahony, 2015; Morales,
2016) showed that breed and fat source interaction had higher effect on Friesian than Jersey
cows when dietary fat was changed. Zebu cows’ milk fat can be as higher as 7%. The strain
and genetic constitution of cows within a breed have a large effect on milk composition. An
average value from many thousands of analyses shows that, Ayrshire breed has higher fat
content than Friesian (Holstein) breed (Table 2). The poorer milk quality from specific herd
of one breed compared with another herd of a different breed can be due to individual
breeding and management. Friesian breed consumed more dry matter (DM) and yielded more
fat corrected milk (FCM) than Jersey cows but percentages of fat were higher in milk yielded

3
by Jersey cows (Beaulieu and Palmquist, 1995). Milk fat from Jersey cows contained a higher
proportion of SCFAs and medium chain fatty acids (MCFAs) and lower proportions of
palmitic and oleic acid than in Holstein milk fat (Beauleu and Palmquist, 2009).

Table 2: milk fat and protein of different dairy cattle breeds

Breed Percent
Total fat Total protein True protein Total solids
Ayrshire 3.88 3.31 3.12 12.69
Brown swiss 3.98 3.52 3.33 12.64
Guernsey 4.46 3.47 3.28 13.76
Holstein 3.64 3.16 2.97 12.24
Jersey 4.64 3.73 3.54 14.04
Milking 3.59 3.26 3.07 12.46
shorthorn

Source: (Neitz and Robertson, 2011)

2.3 Diseases and their medication

Effects of mastitis on milk composition are determined by severity of infection, extent of
infection, secondary physiological changes which alter metabolism of mammary cells and/or
the cow, and whether the epithelial cells are disrupted or milk components are destroyed by
enzymatic action. The fat content from milk of two different farms decreased due to mastitis
infection and due to disruption of mammary gland secretory cells (Kiro and Stefanov, 2006).

2.4 Composition of Milk Fat

The milk fat is composed of 98.5% triglyceride, 0.6% phospholipids, 0.1% monoglyceride,
0.1% free FAs, 0.4% diglyceride, 0.3% sterol (cholesterol and cholesterol esters) and 0.03%
cholesterol (Vanstone and Daugall, 2017). Milk fat globules vary in size from as small as
0.1µ to 22µ in diameter. Majority are within the range 1 – 5 in diameter (Webb, Johnson
and Alford, 2010)

2.4.1 Triglycerides
The main milk fat component is triglycerides which are comprised of a glycerol backbone
bonded to three different FAs. The FAs are composed of a hydrocarbon chain and a carboxyl
group. Milk fat contains both SFAs and UFAs (Lehninger, 2007). LCFAs include C14:0 -
myristic 11%, C16:0 - palmitic 26%, C18:0 - stearic 10% and C18:1 - oleic 20%. SCFAs
(11%) include C4:0 - butyric, C6:0 - caproic, C8:0 - caprylic, and C10:0 - capric. Butyric FA
is specific for milk fat of ruminant animals and is responsible for the rancid flavor when it is

4
cleaved from glycerol by lipase action. Milk fat triglycerides are synthesized in the mammary
epithelial cells. However, the FAs used to synthesize the milk triglycerides may arise from 3
sources namely from breakdown of blood lipids, dietary origin and from de novo synthesis
within the mammary epithelial cells (Feingold et al., 2013)

2.4.1.1 Blood Lipids

40% to 60% of the triglycerides fatty acids come from the blood. They are derived from very
low-density lipoproteins (VLDL) composed of 90 to 95% lipids, 55 to 60% triglycerides on
the inner core and five to 10% proteins at the outer surface. Chylomicrons, containing
ingested FAs from the intestine, act as a source of blood derived FAs for the mammary gland
(Lehninger, 2007). The FAs contained in VLDL and Chylomicrons are dependent upon
dietary lipids and on mobilized fat from body adipose tissues (Stryer, 2015).

2.4.1.2 De novo fatty acid synthesis

The De novo fatty acid synthesis means the synthesis of new FAs from precursors absorbed
from the blood (Lehninger, 2007). De novo synthesis of FAs occurs in the cytoplasm of the
mammary epithelial cells. In the ruminant, the carbon sources used for FAs synthesis are
acetate and β-hydroxybutyrate acid (ßHBA) absorbed from the rumen. Feeding unsaturated
dietary fats resulted in lower concentrations of SCFAs and MCFAs and higher concentrations
of LCFAs in milk fat and butter

2.4.1.3 Cholesterol
Cholesterol is a waxy substance consisting of fats/lipids and proteins. It is present in milk to
the extent of 0.03% (Vanstone and Daugall, 2017). It is formed in the liver and carried in the
blood on the carrier molecule known as lipoproteins which are either the VLDL or low-
density lipoprotein (LDL) or high-density lipoprotein (HDL).

5
 CHAPTER THREE

3.0 MATERIALS AND METHODOLOGY

3.1 Description of The Study Area

The experiment will be conducted at the Department of Animal, Aquaculture and Range
management Sciences (DAARS) MILK LABORATORY and at Magadu dairy farm area at
Sokoine University of Agriculture (SUA) in Morogoro Tanzania.

The university is located between 60 and 7° S and 37o and 38° E and altitude of 500 to 600 m
above sea level, 3 km far away from Morogoro town the area receives an average annual
rainfall of between 600 to 1000 mm and experiences day temperature ranging between 20 to
27° C in the coolest months (April to September) and 30 to 37° C during the hottest months
(October to March).

3.2 Animals and Their Managements

Twelve Friesian dairy cattle will be selected from the herd of 24 active milking Friesian
herd found at magadu dairy farm.
The selection of these animals will be based on the following criteria. These selection
criteria for selection of the animals are as follows:
 Body condition score, the animal to be selected should have good physical look like their
size, their standing posture which will help to determine their heath
 good health with no history of recent clinical diseases.

Before the start of the milk collection the animals will be checked for their health status
i.e., free from diseases. The animals will be given the same quantity of feed(ration) of
similar composition.

3.2.1 Animal Feeding

The animals were allowed to graze during the day time for about 5 hours from 0800 hours to
1300 hours. It was assumed that the intake from grazing will supply 1% of BW dry matter
intake (DMI). From 1300 to 1500 hours animals were allowed to rest in a resting paddock
that was supplied with clean water troughs for animals to drink. Animals were supplied with
3 kg of treatment concentrate diets at each milking that was done twice per day at 0600 and
1600 hours. The amount of concentrate offered was assumed to supply 1.2% of BW DMI.

6
The following is a table showing the standard formulation of supplementary diet at Magadu
dairy farm
Table 5: Feed formulation and nutrient composition
Ingredients Amount (%)
Maize bran (MB) 56.40
Cotton seed cake (CSC) 18.30
Sunflower seed cake (SSC) 23.25
Premix 0.50
Limestone 1.00
Salt 0.50
Crude protein (CP) 15.88
Metabolizable Energy (ME) 13.79

3.3 Milk Collection and laboratory analysis

A total of 12 Friesian dairy cattle found at magadu dairy farm was used in this study where
by the evaluation took for over three weeks where by one analysis activity was done once per
each week. In each evaluation day the 12 animals were milked by machines and their milk
was analyzed on the laboratory. I will take half a liter from each of these 12 cattle and then
mix the milk to obtain 6 liters whereby from the sample will be composed for the laboratory
analysis.
The milk sample will be analyzed for the milk density, milk pH, milk protein, buffer fat, total
solids (TS), (SNF). Milk pH will be determined by milk pH meter, milk density by
(lactometer test) by using hydrometer (ILCA, 1988); BF will be determined by using Gerber
method (ILCA, 1988); N percent will be determined by Kjeldahl method outlined by
Bremner and Mulveney (1992) and the CP percent will be calculated by the formula: CP% =
N% x 6.38, where;

3.3.1. Determination of Total Nitrogen Content in Milk/Milk Product by Kjeldahl

Method
This method employs the use of traditional Kjeldahl apparatus using Kjeldahl flask.
Reagents
A. Potassium sulfate (K2SO4): Nitrogen free or low in nitrogen content.
B. Copper (II) sulfate solution: Dissolve 5.0 g of copper (II) sulfate pentahydrate
(CuSO4.5H2O) in water and make up the final volume to 100 ml in a 100 ml volumetric
flask.

7
C. Concentrated sulphuric acid: At least 95 - 98% (m/m), nitrogen free, ρ20 approximately =
1.84 g/ml
D. Sodium hydroxide solution, 50%, m/m (low in nitrogen): 50 g NaOH pellets was
dissolved in water and finally made to 100 g.
E. Indicator solution: 0.1 g of methyl red was dissolved in 95% (v/v) ethanol and diluted to
50 ml with ethanol. 0.5 g of bromocresol green was dissolved in 95% (v/v) ethanol and
diluted to 250 ml with ethanol. 1 part of methyl red solution was mixed with 5 parts of
bromocresol green solution.
F. Boric acid solution (H3BO3): 40 g of boric acid was dissolved in hot water, the solution
cooled and diluted to 1 L. 3 ml of methyl red - bromocresol indicator solution was added,
mixed and the solution was stored in borosilicate glass bottle. The solution turned to light
orange. The solution was Protected from light and sources of ammonia fume during storage.
G. Standard Hydrochloric acid solution: 0.1 ± 0.0005 N.
H. Ammonium sulfate [(NH4)2SO4]: Minimum assay 99.9% on dried material. Immediately
before used the ammonium sulfate dried at 102 ± 2°C for not less than 2 h. Cooled to room
temperature in a desiccator.
I. Tryptophan (C11H12N2O2) or Lysine hydrochloride (C6H15ClN2O2): Minimum assay 99%.
J. Sucrose with a nitrogen content of not more than 0.002% (m/m).

Apparatus
A. Kjeldahl flasks: Kjeldahl, hard, moderately thick, well-annealed glass, 500 or 800 ml
capacity.
B. Distillation flask: Same Kjeldahl flask as in 1, fitted with rubber stopper through which
passes lower end of sufficient rubber bulb or trap to prevent mechanical carryover of NaOH
during distillation. upper end of the bulb was Connected to condenser tube by rubber tubing.
graduated 500 ml Erlenmeyer titration flask was Used to collect the distillate. The outlet of
condenser was traped in manner to ensure complete absorption of ammonia distilled into
boric acid solution.
C. Digestion apparatus: To hold the Kjeldahl flasks in an inclined position approximately
45°C with electric heater or gas burners that do not heat the flasks above the level of their
contents, and with a fume extraction system. The heater source should be adjustable to
determine the maximum heater setting to be used during digestion. the heat source was
preheated at the heater setting for evaluation. In the case of a gas heater, the preheat period
was 10 min and for an electric heater the preheat period could be 30 min. the heater setting
8
that brings 250 ml of water including 5 – 10 boiling aids was determined with an initial
temperature of 25°C to a rolling boil in 5 – 6 min for each type of heaters. This was the
maximum heater setting that was used during digestion.
D. Conical or Erlenmeyer flask: 500 ml capacity, graduated at every 200 ml.
E. Burette: 50 ml capacity, graduated at least at every 0.1 ml.
F. Boiling aid: Mesh size 10 high purity amphoteric alundum granules, plain.
G. Measuring cylinders: 50, 100 and 500 ml capacities, graduated

Procedure
Test Portion and Pre-treatment
To the clean and dry Kjeldahl flask, 5 – 10 boiling aids, 15 g K 2SO4, 1.0 ml of the copper
sulfate solution, approximately 5 ± 0.1 g of prepared milk sample was added, weighed to the
nearest 0.1 mg, and 25 ml of concentrated sulfuric acid was added. 25 ml acid was used also
to wash down any copper sulfate solution, K 2SO4 or milk left on the neck of the flask. Gently
the contents of the Kjeldahl flask were mixed.

Determination
Digestion
A. The fume extraction system of the digestion apparatus was turned on prior to beginning of
the digestion. the Kjeldahl flask was heated plus its contents on the digestion apparatus using
a heater set at low enough such that charred digest does not foam up the neck of the Kjeldahl
flask. Digestion at this heat was done for 20 min until white fumes appear in the flask. the
heater setting was Increased to half way to the maximum setting as determined and continued
the heating period for 15 min. At the end of 15 min period, the heat was increased to
maximum setting.
B. After the digest cleared (clear with light blue-green colour), then continuous boiling for 1
h to 1.5 h at maximum setting. The total digestion time was 2 hours. Note: To determine the
specific boiling time required for analysis conditions in a particular laboratory using a
particular set of apparatus, for milk analysis, select a highprotein, high-fat milk sample and
determine its protein content using different boil times (1 h – 1.5 h) after clearing. The mean
protein result increases with increasing boil time, becomes consistent and then decreases
when boil time is too long. Select the boil time that yields the maximum protein result.
C. At the end of digestion, the digest was clear and free of undigested material. the acid
digest was cooled to room temperature over a period of approximately 25 min. If the flasks
9
are left on hot burners to cool, it will take longer to reach room temperature. The cooled
digest turned into liquid at the bottom of the flask at the end of 25 min cooling period. Do not
leave the undiluted digest in the flask overnight. The undiluted digest may crystallize during
this period and it will be very difficult to get that back into the solution to avoid this situation.
Note: Excessive crystallization after 25 min is the result of undue acid loss during digestion
and can result in low test values. Undue acid loss is caused by excessive fume aspiration or
an excessively long digestion time caused by an incorrect maximum burner setting.
D. After the digest is cooled to room temperature, 300 ml of water was added to 500 ml
Kjeldahl flask or 400 ml of water when using 800 ml Kjeldahl flask. The neck of the flask
was washed too with water. The contents were mixed thoroughly ensuring that any crystals
which separate out are dissolved. Then 5 - 10 boiling aids were added. The mixture was
cooled again to room temperature prior to the distillation.

Distillation
A. The condenser water for the distillation apparatus was turned on. 75 ml of 50% (m/m)
sodium hydroxide solutionwas added to the diluted digest by carefully pouring the solution
down the inclined neck of the Kjeldahl flask, so as to form a clear layer at the bottom of the
bulb of the flask.
B. Immediately after the addition of sodium hydroxide solution to the Kjeldahl flask, it was
connected to the distillation apparatus, the tip of whose condenser outlet tube is immersed in
50 ml of boric acid solution with indicator contained in a 500 ml Erlenmeyer flask.
Vigorously swirl ed the Kjeldahl flask for mix its contents thoroughly until no separate layers
of solution were visible in the flask any more. the flask down on the burner was set. The
burner was turned on to a setting high enough to boil the mixture. Distillation Continued until
irregular boiling (bumping) starts and then immediately disconnected the Kjeldahl flask was
turned off the burner. Also, the condenser water was turned off.
C. The distillation rate was approximately 150 ml, distillate is collected when irregular
boiling (bumping) started and the volume of the contents of the conical flask was
approximately 200 ml. The efficiency of the condenser was such that the temperature of the
contents of conical flask does not exceed 35°C during distillation.

10
Titration
Boric acid receiving solution was titrated with standard hydrochloric acid solution (0.1 N) to
the first trace of pink colour. Take the burette reading to at least the nearest 0.05 ml. A
lighted stir plate was aiding visualization of the end point.

Blank Test
Simultaneously carry out a blank test by following the procedure as described above taking
all the reagents and replacing the milk sample with 5 ml water and about 0.85 g of sucrose.

Note:
1. The purpose of sucrose in a blank or a recovery standard is to act as organic material
to consume an amount of sulfuric acid during digestion that is roughly equivalent to a
test portion. If the amount of residual free sulfuric acid at the end of digestion is too
low, the recovery of nitrogen by both recovery tests (See Section 19.1.1.3.4. i.e.,
Nitrogen recovery test) will be low. If the amount of residual acid present at the end
of the digestion is sufficient to retain all the nitrogen, but the temperature and time
conditions during digestion were not sufficient to release all the nitrogen from a
sample, then the nitrogen recovery will be acceptable as per Section

2. The amount of titrant used in the blank should always be greater than 0.00 ml. Blanks
within the same laboratory should be consistent across time. If the blank is already
pink before the beginning of titration, something is wrong. Usually, in such cases, the
conical flasks are not clean or water from the air that may condense on the outside of
the condenser apparatus has dripped down into the collection flask to cause the
contamination.

Calculations
The nitrogen content, expressed is a percentage by mass, by following formula
Wn = 1.4007 x Vs − VB x N W
Wn = nitrogen content of sample, expressed as a percentage by mass;
VS = volume in ml of the standard hydrochloric acid used for sample;
VB = volume in ml of the standard hydrochloric acid used for blank test;
N = Normality of the standard hydrochloric acid expressed to four decimal places;
W = mass of test portion in g, expressed to nearest 0.1 mg.
11
Calculation of Crude Protein Content
The crude protein content, expressed as a percentage by mass, is obtained by multiplying the
nitrogen content by 6.38. Express the crude protein results to three decimal places
CP% = N% x 6.38

3.3.2. Determination of Fat in Milk

Reagents /Apparatus

A. Sulphuric acid with a density of 1.807 g/ml at 27°C corresponding to a concentration of

sulphuric acid from 90-91 percent by weight.

B. Amyl alcohol for milk testing (furfural free). of density between 0.816 g/ml at 27°C.

C. Gerber sulphuric acid: Sulphuric acid with a density of 1.812 g/ml at 27°C corresponding
to a concentration of sulphuric acid from 90 to 91% by mass.

Preparation of Gerber Sulfuric acid:

Required volume of water was taken in a Pyrex flask (generally 100 ml of water is required
for 900 ml of concentrated sulfuric acid) kept in a basin of ice-cold water. Carefully the
commercial sulfuric acid was added in small quantities at a time keeping the container
sufficiently cold and gently mixed.

D. Iso-amyl alcohol (C5H11OH): The iso-amyl alcohol of density of 0.805 g/ml at 27°C and
was furfural free.

E. Gerber butyrometer; 6, 8 and 10 percent (ISI marked).

F. Pipette: 10 ± 0.25 ml or automatic measure or tilt measure for sulphuric acid.

G. Pipette: 10.75 ± 0.03 ml for milk.

H. Pipette: 1 ± 0.05 ml or automatic measure or tilt measure for iso-amyl alcohol.

I. Lock stoppers for butyrometers.

J. Lock stopper key.

K. Water-bath: The water-bath was made of a suitable material (e.g., stainless steel). It was
capable of being maintained at 65 ± 2°C and was of sufficient depth as to support the
butyrometer in vertical position with their scale completely immersed. The bath was fitted

12
with horizontal perforated plates to hold the butyrometers and also carried a suitable
thermometer.

Procedures

10 ml of sulphuric acid was Measured and put into a butyrometer tube, preferably by use of
an automatic dispenser, without wetting the neck of the tube. The milk sample was gently
mixed but thoroughly and filled the milk pipette above the graduation line. The outside of the
pipette was wiped and the milk level was allowed to fall so that the top of meniscus is at level
with the mark. The milk was Run into the butyrometer tube along the side wall without
wetting the neck, then left to drain for three seconds and touch the pipette's tip once against
the base of the neck of the butyrometer tube. 1 ml of Amyl alcohol was added, closed with a
lock stopper, shake until homogeneous, inverted for complete admixture of the acid. Kept in
a water bath for 5 minutes. at 65±2°C taking care to have casein particles if any to dissolve
fully, and centrifuge for 4 min. at 1100 rpm. The tubes were put in centrifuge, so as to
conform to radial symmetry, and as evenly spaced as possible, in order to protect bearings of
the centrifuge. The centrifuge was allowed to come to rest. The butyrometer tubes was
removed and place in water bath for 5 min. at 65±2°C. Then observed the percentage of fat
after adjusting the height in the tube as necessary by movements of the lock stopper with the
key. Note the scale reading corresponding to the lowest point of the fat meniscus and the
surface of separation of the fat and acid. When readings are being taken hold the butyrometer
with the graduated portion vertical, keep the point being read in level with the eye, and then
read the butyrometer to the nearest half of the smallest scale division.

3.4 STATICAL ANALYSIS

Descriptive statistics mainly the MEAN/AVERAGE level or percentage of the milk

components (Milk Fat, Milk Crude Protein, Total Solids, Solid Not Fat) and the physio-
chemical properties (milk pH, milk density,) was analyzed. Also, the milking day was
analyzed if they had any significance for the differences in the milk composition using simple
t-test.

The model for t-test is : Yij = µ + Ti +eij

13
 CHAPTER FOUR

4.0. RESULT AND DISCUSION

The followings were the result of the laboratory chemical analysis and further calculations
were done to obtain the all-chemical components of the milk.

CP% = N% x 6.38,
where;
N% = Nitrogen percent.
CP% = crude protein percent.
6.38 = conversion factor for milk.
TS were calculated using the standard formula (ILCA, 1988) as:
L/4 + (1.22 x BF percent) + 0.72, where;
L = lactometer reading.
SNF was computed from the equation (ILCA, 1988);
SNF% = TS% - BF% where;
SNF% = Solid not fat percent.
TS% = total solid percent.
BF% = butter fat percent.

14.01 ×(Titer value−Blank mass)mls × Normality /Molarity ×100

%N =
Sample volume × 1000

Blank mass=0.04mls

Normality =0.1167N

Sample volume=1ml

1. The first week laboratory chemical analysis results

PROPERTY PROPORTION
Titer value 3.53mls
N 0.5706%
BF 5%
CP 3.6404%
PH 6.7
TSF 14.455%
SNF 9.445%
Density 30.5g/cc
14
 2. The second week laboratory chemical analysis results

PROPERTY PROPORTION
Titer value 3.14mls
N 0.5068%
BF 4.6%
CP 3.2334%
PH 6.7
TSF 13.732%
SNF 9.132%
Density 29.6g/cc

3. The third week laboratory chemical analysis results

PROPERTY PROPORTION
Titer value 3.09mls
N 0.4987%
BF 4.3%
CP 3.1815%
PH 6.8
TSF 13.166%
SNF 8.866%
Density 28.8g/cc

The mean proportion/level of the milk components

PROPERTY AVERAGE PROPORTION

N 0.5253mls
BF 4.6%
CP 3.3517%
PH 6.7
TSF 13.7843%
SNF 9.1476%
Density 29.63g/cc

The average was done by dividing the summation of each chemical composition proportion of
a given component for the three weeks by three.

15
 CHAPTER FIVE

5.0 CONCLUSION AND RECCOMENDATIONS

5.1 Conclusion
The average of these milk components and their physio-chemical properties was obtained as
elaborated on the table as shown above. The average of these components was analyzed and
found that the effect of milking day has no effect on these milk components and their physio-
chemical properties on the milk. These values obtained are not constant as can be influenced
by diet, season, breed, diseases and stage of lactation, but for this study most of those factors
like breed, diet, season, stage of lactation, animal health was made uniform so as to reduce
those errors. The variation in milk protein and butter fat is great enough that can be selected
in genetic programs as there is higher correlation between phenotypic characters and the
genotype composition.

5.2 Recommendations
Repeated lab analysis and Records should be taken so as to know the state of our dairy cattle
at magadu dairy farm in terms of milk quality so as the data can be available to anyone in-
need with them for present and future use as records are very important in any animal
production since it will be used at any time for a particular issue.
Also, in order to maintain these components values/percentage, the management practices
upon the animals and their surroundings needs to be followed strictly so as to avoid the drop
of their genetic potentials due to poor management.

16
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