Comparative Analysis of Goat Milk and Colostrum

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.
SCIENCE COLLEGE WAHDAT

ROAD LAHORE.PAKISTAN.39250.
WWW.MINIPRODUCTIONS.TK
+923234666709. WWW.MINIPRODUCTIONS.TK
A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT


ACKNOWLEDGEMENT
Bundle of thanks to Allah the almighty Who enabled me to perform this research work. His countless blessings enabled me to complete my research work successfully. All respect for the last Holy prophet Mohammad (P.B.U.H) who (P.B.U.H) forever is a touch of knowledge and kindness for humanity. I submit my deepest gratitude to my respected, most loved research supervisor Prof. Dr. Asif Mehmood Qureshi, of Zoology Department of Govt. College of Science, Wahdat Road, Lahore for his scholarly guidance, encouraging attitude and enlightened views. His nice and scholarly point of view was the real source of inspiration for me during my studies. Thanks once again to him for providing me necessary facilities for my research work. Experimental work for my research thesis was carried out at Food and Biotechnology Research center (FBRC) of PCSIR Laboratories, Complex Lahore for which I am thankful to authorities. I feel rejoice to acknowledge with gratitude to Dr .Abdul Majeed Sulariya, Principal Scientific Officer (PCSIR Laboratories, Lahore) for his inspiring guidance and
indispensable suggestions. I especially want to acknowledge about his nice way of telling the correct procedures during my experimental work whenever I forgot something. Under his custody my research work was performed very nicely, So I want to pay a big thanks to him for his enlightened, and encouraging attitude to me. I also pay my regard to Abdul Aahad Rashid, the scientific officer in FBRC of PCSIR Laboratories Complex Lahore, Mrs. Shamim Ijaz and Mrs. Shabana as they provided me their valuable suggestions and guidance to complete my all experimental work in laboratory. I pay my best regard to my father and my mother whose financial and passionate encouragement made it possible to complete this work. I would like to thank my friends especially M.Shaheryar Hassan, Junaid Ahmed, Rifat Perven, and all my class fellows and colleagues for their cooperation. Special thank to the laboratory staff of PCSIR LABS for their help during my research.
SARWAR ALLAH DITTA.

Dedicated To My Mother The Most Perfect Lady Of The World

Abstract
Goat colostrum (three days postpartum) was examined for the crude proteins, fats, ash, moisture, lactose, and total solids. Mature milk was taken as control. During this study it was analyzed that fat, proteins, ash, and total solids were much higher in the first day colostrum, and decreased thereafter too much extent in the second and third day colostrum respectively. However the lactose and moisture were lower in the first day colostrum, but latter on these constituents increased in second, and third day colostrum. The concentrations of the major colostrum constituents changed
significantly after birth, the level of different constituents on the third day was very similar to those of the mature milk.

Table of Contents
Contents------------------------------------------------------------------------------------page CHAPTER NO: 1. INTRODUCTION 1 1.1. Composition of the colostrum. 2 1.1.1. Proteins 3 1.1.2. Lipids 8 1.1.3. Enzymes 9 1.1.4. Minerals and Salts 10 1.2. Significance of the colostrum 10 1.3. Immunity 11 1.4. Mechanism of immunity 14 1.5. Objectives of the study. 15 CHAPTER NO: 2. MATERIALS & METHODS 16 2.1. Materials 16 2.2. List of Apparatus 16 2.3. List of Reagents & Solutions 17 2.4. Principles of tests 17 2.5. Determination of Protein Content (Kjeldhal Method) 18 2.6. Determination of Moisture Contents (Oven Drying Method) 19 2.7. Determination of Ash Contents 20 2.8. Determination of Fat Contents (Butyrometer Method) 20 2.9. Determination of Lactose in Colostrum and Milk of Goat 20 CHAPTER NO: 3. RESULTS 22 CHAPTER NO: 4. DISCUSSION 35 CHAPTER NO: 5. REFERENCES 41

CHAPTER NO.1
INTRODUCTION

1. INTRODUCTION
Colostrum is of great nutritional and immunological values for the newborn mammals. It is the first milking of the mammals after giving birth to young one and continues about till three days in goats. The formation of the colostrum in the mammary glands of the mammals starts before the delivery of the neonates, but it comes out of the breast just after the delivery of the neonates. It continues through out the early days of the breast feeding. This special kind of milk is yellow to orange in color, which is thick and sticky in nature. It is the first milking consumed by the newborn, and which is formed and stored in the mammary glands during the late pregnancy (Linzell and Peaker, 1974). Lambs are born hypo-immunocompetent, and at the time of birth they have a small store of energy for the heat production and metabolism (Odoherty and Grosby, 1997). The capacity of the lamb to utilize food can affect its growth performance (Emsen et al., 2004). The most satisfactory way of providing the newborn with the passive immunity against diseases is to ensure that it gets a large quantity of good quality colostrum in the early days of life (Napolitano et al., 2002). Colostrum meet all the nutritional requirements of the newborn (Blum and Hammon, 2000;Blum and Baumrucker, 2002). So it is required to feed neonates with high quality colostrum as soon as possible after birth to decrease the diseases susceptibility and neonatal mortality (Wittum and Perino, 1995; Tyler et al., 1999). Feeding a good deal of fresh colostrum within the first few hours of birth plays a vital role in neonates health, survival, and subsequent performance (Robison et al., 1988; Wittum and Perino, 1995).

1.1 Composition of the colostrum

Colostrum contains more proteins, sodium, chloride, and less potassium and lactose than milk secreted in an established lactation. It also contains vitamins, hormones, growth-factors, cytokines, enzymes, and other bioactivepeptides (Swaisgood, 1995). In addition colostrum contains high concentrations of immunoglobulins, and it has been confirmed that it provides maternal immunity to the young in a number of species of the mammals (Linzell and Peaker, 1970). The contents of the postpartum milk deviate widely from the normal milk of the organism in contents like, immunoglobulins, growth-factors, proteins, non-proteineous-nitrogen, ash, minerals, and vitamins level ( Rauprich et al., 2000; Blattler et al., 2001). Colostrum composition and physical properties change which mainly depend on various factors including the animal age, number of lactation cycles, breed, diet, and diseases (White and Davies, 1958a, b, c; Cerbulis and Farrell, 1976; Donnelly and Horne, 1986; Horne et al., 1986; Rodriguez et al., 2001). It has been reported that the pH of the colostrum is lower (Edelsten, 1988), the specific gravity slightly higher (Haggag et al., 1991), and the immunoglobulins content about 100 times greater than that of the mature milk (Renner et al., 1989), it has been estimated that constituents such as Ca, Mg, P, Na, and K are much greater at the first milking postpartum (Klimes et al., 1986) and also contain a good deal of metabolities derived from alveolar epithelial cells ( Peaker and Linzell, 1975), and immune-cpmpetent cells (Lee et al., 1980). Milk composition in the mammals is greatly influenced by the breed, parity, age, nutrition, non-lactating periods, and health status of the animal (Pritchett et al., 1991; Porto et al., 2007). The composition of the colostrum can be discussed under following major headings:

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1.1.1. Protein
1.1.1.1 Immunoglobulins: These are also called antibodies, because these neutralize the toxin in the lymph and circulatory system and provide passive immunity to the neonate against many diseases (Foley and Otterby, 1978). Immunoglobulins provide passive immunity against many diseases of young neonates. Colostrum contains mainly three types of immunoglobulins, i.e IgG, IgM, and IgA.Where IgG accounts for more than 75% of the total immunoglobulins (Korhonen et al.,2000),IgM accounts for more than 5%, and IgA accounts for nearly about 7% of the total immunoglobulins (Devery-Pocius and Larson, 1983). 1.1.1.2 Leukocytes: Leukocytes are the white blood cells that play very important role in fighting against the infections that can harm the neonates (Johnson et al., 2007). These help in clearing the toxic materials which are produced by the invading organisms, and also stimulate the production of a large amount of interferon which interferes with the reproduction of the viruses. Therefore leukocytes protect the neonates from many dreadful diseases (Davis and Drackley, 1998). 1.1.1.3 .Lactoferrin: This is an iron-binding protein which aids the body to utilize the iron. It is well known that iron deficiency can cause anaemic problems. Lactoferrin is also a powerful antiinflammatory agent, which reduces the inflammation accompanied with many health problems (Neville and Zhang, 2000).

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1.1.1.4 Lysozymes: Lysozymes play a key role in the first line of defense against bacteria. It can break down the outer cell wall of certain bacterial organisms, thus inhibiting their reproduction. 1.1.1.5. Enzymes including Peroxidase: The enzymes Peroxidase generate the release of hydrogen peroxide to "burn" or hydrolyze bacteria. This is the basis for the use of hydrogen peroxide as therapy to kill the bacteria. 1.1.1.6. Proline-rich polypeptide (PRP): Proline-rich polypeptide is a hormone which helps to regulate the immune system, keeping it in balance between under and over activity. This is extremely important for those mammals which have auto-immune diseases because it keeps the immune system in balance (Stelwagan et al., 2009). 1.1.1.7. Cytokines: Cytokines are small soluble glycoproteins that act in autocrine-paracrine fashions by binding to specific cellular receptors, operating in networks, and orchestrating immune system development and functions (Bilal et al., 2005). Cytokines are proteins that are involved in the regulation of the immune response. When these proteins are produced by lymphocytes, they are referred to generically as lymphokines (Dumonde et al., 1969), and when they are produced by monocytes or macrophages, they may be called monokines (Frank, 1991). The term interleukin is applied to proteins that function as communicators between leukocytes (Bilal et al., 2005). These cytokines are present at picogram quantities. However, early milk has an abundance of cytokines at a time when neonatal organ systems are immature, suggesting that these bioactive components of milk might be important in neonatal

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development (Oddy et al., 2003; Wallace et al., 1997). Cytokines influence many different aspects in the maturation and
differentiation of cells of the immune system and in the host's response to disease. Cytokines have a broad influence on the immune system and are involved in the production of T-cells, lymph activity, and in regulating the force and duration of an immune response (Frank, 1991). 1.1.1.8. Lymphokines: Lymphokines are hormone-like peptides which are released by stimulated white blood cells. They help to regulate the immune response (Frank, 1991). 1.1.1.9. Growth Hormone (GH): Growth hormone is considered to be an immuno-stimulant because it helps the body to produce antibodies, T-cells and white blood cells. It is particularly important in regeneration of lost muscle mass during illness and has been shown to aid in the growth of the thymus gland. Growth Hormone affects neurotransmitters in the brain improving moods and mental acuity (Roffler et al., 2003). 1.1.1.10. Insulin-like growth factors (IgF) I and II: Insulin-like growth factors (IGF-I and IGF-II) comprise the principal growth factors in milk, and can be found in all mammalian species. Insulin-like growth factor I is a mitogenic
polypeptide, the molecular structure of which is quite similar to that of insulin. This compound stimulates growth, differentiation, and metabolism in a variety of cell types, acting via IGF-I receptors (Zapf et al., 1984; Rechler and Brown, 1988).IGF-I in colostrum may be partially responsible for these effects. Insulin-like growth factor I

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content in bovine and porcine milk has been reported to be in the range of 22 to 26 ng/mL (Collier et al., 1991), and 1.27 to 8.10 ng/mL (Donovan et al., 1994) respectively. The concentration of IGF-I in bovine colostrum showed wide variation. The IGF-I concentration was shown to increase in the final period of pregnancy (Donovan et al., 1994), and served an important function in the development of the postnatal gastrointestinal tract (Philipps et al., 1997). In this regard, supplementation with milk-borne IGF-I may prove to be therapeutic with regard to growth retardation in preterm infants. Insulin-like growth factors improve the function of Growth Hormones and are responsible for many anti-aging and regenerative effects (Baumrucker and Blum, 1994). 1.1.1.11. Epithelial growth factor: Milk contains an abundance of physiologically active proteins that defend against infections (Briese et al., 1986; Brown et al., 1990; Davies., 1989) that are associated with nutrition and metabolic control (Aynsley-Green., 1989; Azuma et al., 1989). Human milk or bovine colostrum in place of serum in cell cultures supports normal growth of various cell types, such as epithelial cells, fibroblasts, and smooth muscle cells (Khar, 1983; Klagsbrun and Neumann, 1979). Physiologically active substances have been found in trace amounts at specific stages during lactation, epidermal growth factor (Petrides et al.,1985), insulin-like growth factor (Collier et al.,1991)), growth factor-like growth factor derived from platelets (Shing and Klagsbrun, 1987), transforming growth

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factor-a (Okada et al.,1991), and transforming growth factor-b (Jin et al.,1991; Cox and Buerk, 1991).Epithelial growth factor stimulates normal skin growth and works on the mucosal surfaces of our bodies. Transforming growth factors A and B are helpful in healing wounds and in the synthesis and repair of DNA and RNA (Grutter and Blum 1991a, 1991b). 1.1.1.12. Fibroblast growth factor: Fibroblast growth factor is the substance which stimulates the growth of new blood vessels and contributes to tissue development, and wound healing (Ronge and Blum, 1988; Esch et al., 1985). 1.1.1.13. Platelet-derived growth factor : Platelet-derived growth factors are involved in the healing of vascular wounds. They are released in conjunction with blood clotting during the healing process (Roffler et al., 2003). 1.1.1.14. Amino acids & inhibitors: Colostrum has been suggested to be an important source of free amino acids for suckling animals (Bengtsson, 1972). It has been known that colostrum contains high activities of different types of inhibitors. These inhibitors are important in the prevention & healing of the gastric ulcer. These inhibitors also limit the breakdown of intake protein (Laskowski et al. 1957). Although the presence of protease inhibitors in colostrum has important immunological implications (Butler, 1974).These inhibitors minimize the potential of colostrum proteins as a source of free amino acids for neonates. Trypsin inhibitors and other protease inhibitors prevent the destruction of immune factors and growth-factors by enzymes (Laskowski et al., 1957). Bovine colostrum contains a large amount of trypsin inhibitor (TI), which

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may assist in transfer of immunity to the neonate (Sandholm and Honkanen-Buzalski, 1979; Jensen and Pedersen, 1982).
1.1. 2. Lipids:
In the goat milk and colostrum following main types of lipids can be easily identified. 1.1.2.1. Fatty Acids: A number of workers in the past decade have reported the analysis of the fatty acids components of the goat milk and colostrum. Most common fatty acid in the goat milk is the triglycerides. It was earlier estimated that the contents of the short and medium chain acids (C4-C14) in the goat clostral fat is slightly higher than the fat of the mature milk (Klobasa and Senft, 1970). Several studies have dealt with the influence of dietary fat on the fatty acids composition of the goat milk fats. Lowering fat intake of the goat from 1g/Kg to 4g/Kg per day depressed milk productions of the fat percentage, and yield of milk fat (Delage and Fehr, 1967). 1.1.2.2. Triglycerides: Colostral and milk fats of cow, sheep, and goat differ slightly in the pattern of the acyl carbon numbers. The distribution of the molecular weights deviates significantly from the molecular weights deviate significantly from that expected if the fatty acids were incorporated randomly into the triglycerides (Jenness, 1980). 1.1.2.3. Phospholipids and Cerebrosides: In the colostrum and milk of the goat, there are present nearly about all type of phospholipids e.g.Phophatidyl-Cholines,Phosphatidylethanolamines,Phosphatidyl-serines, phophatidyl-inositole and
Sphingomylein (Patton and Keenan, 1971). Cerebrosides, glucosyl-

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ceramide and lactosyl-ceramide are distributed between fat globule and skim milk fractions of the goat milk (Kayser and Patton, 1970). 1.1.2.4. Cholesterol and its Esters: Cholesterol content of the goat milk is usually in the range 10 to 20mg/100ml (Steger, 1961). The content of the cholesterol on the colostrums is much greater than milk i.e. 40mg/100ml (Keenan and patton, 1970). By far the greatest portions of the cholesterol in the goat milk is in the free state, only a small fraction is present in the form of ester (Jenness.,1980).
1.1.3. Enzymes:
Two enzymes that probably are involved in the synthesis of glycoproteins have been found in the goat colostrums, and in partially purified form they have been identified in the goat colostrum (Jenness, 1980). One of these enzymes catalyzes the transfer of N-
acetylglucosamine from Uridine diphosphate N-acetyglucosamine to glycoprotein (Johnston et al., 1966). However the second is a soluble sialyl-transferase which transfers sialic acid (Nuraminic acid derivatives) from cytidine monophosphate-sialic acid to lactose or N-
acetyllactosamine, it was purified partially from goat, cow, and human colostrums (Barhthdomew et al., 1973). There are many other enzymes present in the goat colostrum and milk.
1.1.4. Minerals and Salts:

Colostrum contains more sodium, chloride and less potassium (Linzell and Peaker, 1974), but in the goat milk potassium and chloride is higher in the concentrations than the colostrum. Konar et al (1971) demonstrated a negative correlation between K and lactose for the milks of cow, goat, sheep, and swine from their own work and published data

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from few other species. Chloride is correlated positively with K and negatively with lactose, but Na is not correlated significantly with Cl, K, or lactose (Jenness, 1980). Citrate is a sort of harbinger of lactogenesis in the goats (Fleet et al., 1975; Peaker and Linzell, 1975). Its concentrations in the mammary secretions increase sharply from virtually zero to the normal 150 to 200mg/100, l on the day of parturition (Jenness, 1980). According to the review of Jenness (1980) on the goat milk compositions, milk contains both carbon dioxide and carbonate ions, carbonate ion may reach to 1.9mmoles/liter (Linzell & Peaker, 1974). The total contents of the carbon dioxide and carbonate ion may reach to 3.4mM (Jenness, 1980).
1.2. Significances of the colostrum

Feeding a sufficient quantity of high-quality colostrum within the first few hours of birth plays a vital role in calf health, survival, and subsequent performance, immunity against diseases (Robison et al., 1988; Wittum and Perino, 1995; Faber et al., 2005), and other functions like:(i). It influences the metabolism, endocrine systems and the nutritional status (Guilloteau et al., 1997; Blum and Hammon, 2000). (ii). Ingested colostrum stimulates the development and function of the Gastrointestinal tract (GIT) in the neonates (Blum and Hammon, 2000). (iii). Ingested immunoglobulins, some proteins and enzymes are absorbed in particular during the first hour after the birth (Baumrucker et al., 1994; Hadorn and Blum, 1997). (iv). Ingested growth factors e.g. insulin like growth factors (IGF)-I) and hormones which are the sole components of the colostrum are barely absorbed in the neonate (Baumrucker et al., 1994; Grutter and Blum, 1991).

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1.3. Immunity
The role of the colostrum with reference to the immunity has been extensively studied in the calves (Davis & Drackly, 1998). It provides protection to the immune system of newborn calves and passive immunity against pathogens. The role of colostrum as indispensable nourishment for newborn mammals has been established (Levieux, 1984). Due to its high content in Ig mainly IgG , colostrum provides the major antimicrobial protection and confers passive immunity preventing diseases caused by microbial infections in the newborn (Foley and Otterby, 1978). A number of factors have been identified that influence IgG absorption by the young calf, including the quantity and quality of colostrum, time of first feeding, metabolic status of the calf, and colostrum management practices (Stott et al., 1979a,b; Garry et al., 1996; Morin et al., 1997; Quigley et al., 1998, 2001). Calves with FPT (Failure of Passive Transfer) are more likely to die or become chronically ill (Donovan et al., 1998). When neonates are born they are
agammaglobulinemic (Robison et al., 1988). This condition is a consequence of the absence of Ig transplacental passage. In fact, calves have immature innate defense mechanisms and have no specific immunity at birth. Therefore, passive immunization is crucial and can be provided by ingestion of colostrum containing high levels of Ig (Porto et al., 2007).Thus, it is generally recommended that newborns be fed with fresh colostrum as soon as possible after birth. Colostrum intake supports the
adaptation of calves to environment, and establishes passive immunity (Stott and Fellah, 1983). It also supports development and function of the gastrointestinal (GI) tract, influences metabolic and endocrine systems and neonatal nutritional

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status (Demigne and Remesy, 1984; Simmen et al., 1990; Grutter and Blum, 1991b; Lee et al., 1995; Odle et al., 1996; Blum et al., 1997; Guilloteau et al., 1997; Hadorn and Blum, 1997; Buhler et al., 1998; Hammon and Blum, 1998, 1999). Neonates have underdeveloped digestive and regulatory systems therefore they are greatly challenged by a new environment at birth (Porter and Barratt, 1987). Newborn mammals do not normally consume solid foods, so their mother's milk is the only source of exogenous amino acids for the synthesis of proteins, neurotransmitters, hormones, polyamines, purine and pyrimidine nucleotides, creatine, carnitine, porphyrins, and other biologically important molecules (Reeds, 1988). Some amino acids, such as glycine (Wetzels et al., 1993), histidine (Noronha-Dutra et al., 1993) and taurine (Wright et al., 1986), are effective scavengers of free radicals and therefore may help prevent or alleviate potential intestinal injury. Thus, milk plays a vital role in the survival and growth of mammalian neonates. Colostrum ingestion is the natural process that most significantly confers efficient immune protection to neonatal calves during their initial life stages (Robison et al., 1988). Effective immunity transfer via colostrum ingestion also depends on the ability of the neonate to ingest and absorb the ingested Ig. Absorption of intact large proteins by the intestine of the newborn occurs only within the first 24 h of life (Matte et al., 1982), when IgG becomes increasingly detectable in the calfs blood. Low IgG plasma concentrations are directly related to calf morbidity and mortality (Besser and Gay, 1994), as well as to impairment of the calfs long-term performance (Wittum and Perino, 1995). Plasma IgG concentrations lower than 10 mg/ ml, extended for 24 to 48 h after birth, indicate failure in passive transfer (Arthington et al., 2000), hence neonates are put forward to many risky diseases.

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Failure
of
the
neonatal
calf
to
absorb
adequate
colostral
immunoglobulins into circulation within the first 24 h of life results in increased risk for neonatal disease and mortality and a negative effect on the future health, longevity and performance (Davis and Drackley, 1998; Faber et al., 2005). One of the reasons for the high mortality of newborn calves is their susceptibility to infections. They do not have an available humoral immune system before ingesting colostrum from their mother, their own immune system cannot react effectively with any antigens (Kamada et al., 2007). If calves do not receive a sufficient amount of colostrum, they fall into hypogammaglobulinemia. Suffering calves have high sensitivity to infectious viz diarrhea and pneumonia, so their mortality rate is high. Therefore, the importance of colostral IgG supply is emphasized in animal husbandry; however, various reasons (low colostrum production, low IgG concentration in the colostrum, lack of instinctive suckling by the calf or dam, and physical injury to the calf) may prevent sufficient IgG transfer from the colostrum to calves (Perino et al., 1995; Weaver et al., 2000; Moore et al., 2002).
1.4. Mechanism of immunity:

It is known that pinocytosis of intestinal epithelial cells mediates the active transfer of IgG from the maternal colostrum to newborn calves (Kruse, 1983; Kaup et al., 1996); however, the ability of cells to pinocytose proteins disappears within 24 h after delivery. This means that a delay of providing colostrum also interferes with effective IgG absorption. Ingestion and absorption of colostral immunoglobulins are two of the most important aspects in the preven tion of neonatal calf disease because calves acquire virtually no immunoglobulins in utero. In spite of this knowledge, failure of transfer of passive immunity (FTPI) remains extremely common (Weaver et al., 2000). Calves with inadequate immunoglobulin concentrations have reduced growth rates, increased risk of

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disease and death, increased risk of being culled and decreased milk production in their first lactation (DeNise et al., 1989; Tyler et al., 1998; Tyler et al., 1999; Virtala et al., 1999; Faber et al., 2005). The transfer of immunity from colostrum ingestion is generally considered to be adequate if serum IgG concentrations are above 1,000 mg/dL (McGuirk and Collins, 2004).These include administering a large quantity of good quality colostrum to provide adequate immunoglobulin mass within the first few hours of life(Smith and Foster, 2007). Effective immunity transfer via colostrum ingestion also depends on the ability of the neonate to ingest and absorb the ingested Ig (Porto et al., 2007).Supplying a sufficient amount of IgG to newborn calves is largely dependent on human activities now; however, there is no available technique to increase the efficiency of IgG transfer (Kamada et al., 2007).

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1.5. Objectives of the study

The analysis of the colostrum and nutrient contents will help for the identification of the areas for improvements to increase survival of the dairy goats. In this research work main focus will be on the changes in colostral components with parturition time especially in reference to proteins, fats, ash, moisture, and lactose. Mainly there are two objectives of this study
I.
To analyze different parameters like protein contents, moisture contents, ash, fats, lactose, and total solids in the first three days milking after parturition in the goat.
II.
To make comparative analysis between the mature milk and colostrum constituents in the goat.

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CHAPTER NO.2 MATERIALS AND METHODS

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2. MATERIALS AND METHODS
2.1. Material:
Fresh colostrum samples were collected from a local farm which is situated on the 26 Km- Sheikhupura road Lahore Pakistan. Samples were collected in the sterilized bottles and then stored in refrigerator before carrying to laboratory.
2.2. List of Apparatus

(i) Kjeldhal digestion apparatus (ii) Complete distillation apparatus with heat source (iii) Digestion bulbs and delivery tubes (iv) Pipettes of 50ml (v) Oven (vi) Desiccator (vii) Crucibles (viii) Weighing Balance (ix) Furnace (x) Desiccator (xi) Butyrometer (xii) Pipettes (10.94ml) (xiii) Plastic stoppers (xiv) Centrifuge machine (xv) Burettes (xvi) Beakers (xvii) Hot plate

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2.3. List of Reagents & Solutions

1. HCl (standard solution of N/70) 2. Digestion tablets 3. Sulphuric Acid (concentrated) 4. 40%NaOH standard solution 5. 2% Boric acid 6. Gerber Sulphuric acid :( specific gravity 1.807 to 1.812 at 27C) Transfer 100ml of water in the Pyrex flask keeps in a basin of icecold water. Carefully add concentrated H2SO4 (900ml) in small quantity at a time keeping the container sufficiently cold. Mix gently, cool the flask. After cooling, check the specific gravity of the acid with hydrometer and if necessary adjust the acid to the correct specific gravity with the addition of the water or acid till the specific gravity is in the range of 1.807 to 1.812 at 27C. Store in a glass stopper bottle. 7. Isoamyl-alcohol (95%): clear, colorless liquid shall distill between
130C to 132C and specific gravity shall be 0.803 to 0.805 at 27C distilled water. 8. 6N HCl
9. 20%NaOH 10. Benedicts reagents. 11. Phenolphthalein solution 12. 0.1N NaOH
2.4. Principles of tests

Protein test: This method depends upon the oxidation of organic matter with sulphuric acid in the presence of a catalyst and simultaneous formation of ammonium salts and amines from the nitrogen in the colostrum. The ammonia and the amines may be distilled off when the solution is made alkaline. The distillate is trapped in the standard acid and nitrogen is determined by titration.

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Moisture test: The moisture content of the colostrum can be determined by means of the air oven drying. This is simply done by evaporating all the water and calculated by the difference of the weight of the crucible before and after the drying. Ash test: The ash content of the colostrum can be determined by means of the air furnace. This is simply done by evaporating all the water and ashing of the samples. Fat test: The Gerber sulphuric acid is used to dissolve the casein in milk without charring the fat and liberate the fat, which remains in the liquid state due to heat produced by the acids. Iso-amyl-alcohol, is used to lower down the surface tension in the medium to facilitate the separation of the fats from aqueous phase. On centrifugation the fat being lighter will be separated on the top of the solution.
2.5. Determination of Protein Content (Kjeldhal Method)

I. II. III. IV. V. VI. VII. VIII. IX. X. Take 10gm colostrum in digestion bulb and keep it in oven overnight for drying. Add 18ml of H2SO4 and one digestion tablet in the bulb. Then heat it for 4-5 hours on burner and check so that sample became transparent. Make the volume of sample up to 100ml with distilled water. Take 5ml of sample solution and 10ml of NAOH into the in flask. Then attach the flask in Kjeldhal apparatus for distillation. Take 10ml of boric acid in the beaker and put in the Kjeldhal apparatus. Then on the burner and start distillation. It takes 5-7 minutes until the boric acid becomes colorless. After distillation, titrate the sample against standard acid HCl (0.0142 N) till the end point appeared which is pink. XI. Note the reading as N2 Gas %age.

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XII.
By applying the following formula we can calculate the protein %.
%age N = Titre x 20 5
= (A) mg N
= (A) x 100
= (B) mg N Weight of sample
= (B) 1000
= (C) g N
Protein % age = (C) x Dairy factor (6.38)
= (D) Proteins %
2.6. Determination of Moisture Contents (Oven Drying Method)

I. Take 5gm of colostrum sample in a dried and Pre-weighed crucible. II. Keep the crucibles in oven at 105oC over night. III. Take out the crucibles from oven and cool the crucibles in desiccator. IV. Again weigh the crucibles with dried sample. V. Note the reading as moisture %age.

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VI. The calculations to determine the moisture content of colostrum were made from the following formula:-
Moisture % = wt. of fresh sample wt. of sample after drying x 100 Weight of sample
2.7. Determination of Ash Contents

I. Take 10 gm of colostrum in dried and pre-weighed crucibles II. First of all charred the sample. III. Take out the crucibles from electric furnace and cool the crucibles in desiccator. IV. Again weigh the crucible with sample and note the value as fat %age. V. The calculations were made by the following formula:-
Ash = weight of crucible and ash weight of crucible x 100 Weight of sample

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2.8. Determination of Fat Contents (Butyrometer Method)

I. Take 10ml of sulphuric acid in a butyrometer. II. Take 10.94ml of colostrum sample in the same butyrometer. III. Add 1ml of Isoamyl-alcohol in the butyrometer. IV. Now make volume in the butyrometer scale upto 5 scale adjustment. V. Always prepare duplicate of each sample so do it similarly. VI. Now put the butyrometer in the centrifuge machine. VII. After 7-8 minutes centrifugation put out the butyrometer. VIII. Now carefully notice the oily layer readings on the butyrometer scale. IX. Reading on the scale is the Fat %age.
2.9. Determination of Lactose in Colostrum and milk of the goat

I. II. III. IV. Take 10gm colostrum sample by weighing in the beaker. Add 40ml distilled water and 1ml 6N HCl in the sample. Now boil the sample for 3-5 minutes. Then cool and neutralize it with 20% NaOH (about 7 drops of 10ml pipette). V. VI. VII. Make the volume up to 100ml of sample. Filter it and add it in a burette. Take 5ml Benedicts reagent in the flask and add 45ml distilled water, make the volume up to 50ml. VIII. IX. Keep this to boiling for 3-4 minutes. When boiled, titrate it with burette solution till it appears colorless as end point. X. Note the titre value as lactose % in colostrum sample.

30
CHAPTER NO.3 RESULTS

31
3. RESULTS
3.1: Estimation of Protein contents:
Table 3.1 is presenting the protein contents in first three days goat colostrum and milk. The sample D1 has the protein contents that were estimated for three readings. The protein contents in the first day colostrum (D1) were 14.18%, 14.15%, and 14.17% respectively. Average proteins in D1 sample was 14.17%. The D2 sample which was second day colostrum, in this the protein contents were 7.12%, 7.14%, and 7.11% respectively. Average of the protein contents in the D2 sample was estimated 7.12%. Third day colostrum (D3 sample) has the protein contents 4.53%, 4.50% and 4.53%. Average protein content in the D3 was 4.52%. Sample D (control sample) the mature milk of the goat, was also subjected to
estimations. First reading showed protein content 3.45%, second showed 3.43% of protein contents, and final 3.44% of protein contents. Average protein contents of sample D (control sample) was 3.44%. The colostrum showed decrease in protein contents, from first day colostrum to mature milk. The Figure 3.1 (a) is showing the results of protein contents of different samples D1, D2, D3, and D. The Figure 3.1 (b) is showing comparative analysis of the average protein contents in first three days colostrum and milk. The results revealed that the %age of protein contents are much high in the first day colostrum (14.17%) which was decreased to 3.44% in the mature milk. So there was a sharp decrease in protein contents from colostrum to mature milk.

32
Table 3.1 Days after parturitions D1 D2 D3 D
Protein % in first three days colostrum and milk

Protein % in first three days colostrum and milk
1 14.18 7.12 4.53 3.45
2 14.15 7.14 4.50 3.43
3 14.17 7.11 4.53 3.44
Average % 14.17 7.12 4.52 3.44
Protein% Readings Figure 3.1(a) Protein% in first three days colostrum and milk
Protein % Days Figure 3.1(b) Average protein% in first three days colostrum and milk

33
3.2: Estimation of Fat contents:

Table 3.2 is showing the fat contents in first three days goat colostrum and milk. The sample D1 (first day colostrum) was estimated for three readings. First day colostrum contained 7.9%, 7.7%, and 7.8% fat contents. Average of the fat content in the D1 was 7.8%. Three repeated readings of the fat contents of sample D2 (second day colostrum) were 6.5%, 6.6%, and 6.5%. Average of which was 6.53%. The sample D3 (third day colostrum) had the fat contents 4.7%, 4.9% and 4.8% in three readings. Average value of D3 was 4.8%. Sample D (control sample) showed 3.5% fat in all three readings. Figure 3.2 (a&b) indicated a continuous decrease in fat contents from the D1 (first day colostrum) to D (mature milk), and the fats content of the D3 and D showed much similar values.
Table 3.2 Days after parturitions D1 D2 D3 D
Fat % in first three days colostrum and milk Fat % in first three days colostrum and milk 1 7.9 6.5 4.7 3.5 2 7.7 6.6 4.9 3.5 3 7.8 6.5 4.8 3.5 Average % 7.8 6.53 4.8 3.5

34
Fat% Readings Figure 3.2(a) Fat% in first three days colostrum and milk
Fat % Days Figure 3.2(b) Average Fat% in first three days colostrum and milk

35
3.3: Estimation of Moisture contents:

The moisture contents in first three days colostrum and milk of the goat are shown in Table 3.3. The sample D1 (first day colostrum) showed 73.75%, 73.74%, and 73.77% of the moisture contents (average value 73.74%). Sample D2 (second day colostrum) showed 81.14%, 81.11%, and 81.13%. Average value of sample D2 was 81.13% moisture. Similarly the sample D3 (third day colostrum) showed 84.95%, 84.97% and 84.96% of the fat contents. Average moisture in the sample D3 was 84.96%. Sample D (control sample) showed 88.17%, 88.19%, and 88.13% of moisture contents in three respective readings. Average moisture content of sample D (control sample) was 88.16%. The Fig.3.3 (a&b) is showing the results of D1, D2, D3, and D. The comparison between the moisture contents of different samples showed a large difference in the moisture values. Comparative analysis of the average moisture contents in first three days colostrum and milk showed a continuous increase in the moisture from sample D1 (first day colostrum) to sample D3 (third day colostrum) and moisture values of the mature milk are very close to sample D3 as like shown in Fig.3.3 (a&b).
Table 3.3
Days after parturitions
Moisture % in first three days colostrum and milk

Moisture% in first three days colostrum and milk Average %
D1 D2 D3 D
1 73.75 81.14 84.95 88.17
2 73.74 81.11 84.97 88.19
3 73.77 81.13 84.96 88.13
73.74 81.13 84.96 88.16

36
Moisture% Readings Figure 3.3(a) Moisture% in first three days colostrum and milk
Moisture% Days Figure 3.3(b) Average Moisture% in first three days colostrum and milk

37
3.4: Estimation of Ash contents:

Table 3.4 is showing the ash contents from D1 to D3 and D. The ash contents of sample D1were 1.05%, 1.05%, and 1.05%. Average of sample D1 ash content was 1.05%. The ash contents of sample D2 were 0.96%, 0.95%, and 0.97%. Average of the ash content of D2 sample was 0.96%. Finally the sample D3 has the ash contents of 0.87%, 0.87% and 0.88%. Average ash content in the D3 was 0.87%. Sample D (control sample) showed 0.86%, 0.87%, and 0.87% of ash contents. Average ash content of sample D was 0.87%. The comparative results of samples D1, D2, D3, and D are shown in the Fig.3.4 (a).The Fig.3.4 (b) is showing clear difference between the average ash contents of the sample D1 to D3, and D. The values of ash decreased from day first to third day colostrum. The values of ash contents of third day colostrum and mature milk are very close to each other as shown in Fig.3.4 (b).
Table 3.4
Ash % in first three days colostrum and milk

Ash% in first three days colostrum and milk Average %
D1 D2 D3 D
1 1.05 0.96 0.87 0.86
2 1.05 0.95 0.87 0.87
3 1.05 0.97 0.88 0.87
1.05 0.96 0.87 0.87

38
Ash% Readings Figure 3.4(a) Ash% in first three days colostrum and milk
Ash% Days Figure 3.4(b) Average Ash% in first three days colostrum and milk

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3.5: Estimation of Lactose content:

Table 3.5 is showing the lactose contents in all the samples from D1 to D3, and D. The sample D1 has the lactose contents of 3.0%, 2.9%, and 3.0%. Average of the lactose content of sample D1 was 2.93%. The sample D2 showed 3.8%, 3.8%, and 3.9%. Average of the lactose content in the D2 sample was 3.83%. The sample D3 has the lactose contents 4.4%, 4.5% and 4.5%. Average lactose content in the D3 was 4.43%. Sample D showed 4.9%, 4.7%, and 4.8% of lactose contents. Average lactose content of sample D was 4.8%. The lactose was at low level in the first day colostrum (2.93%), and increased during further days as the time after parturition increased. In the mature milk it was increased to 4.8% as shown in Fig.3.5 (b). The Fig.3.5 (a) showed the results of lactose contents of different samples (D1, D2, D3, and D).
Table 3.5
Lactose % in first three days colostrum and milk

Lactose% in first three days colostrum and milk Average %
D1 D2 D3 D
1 3.0 3.8 4.4 4.9
2 2.9 3.8 4.5 4.7
3 3.0 3.9 4.5 4.8
2.93 3.8 4.43 4.8

40
Lactose% Readings Figure 3.5(a) Lactose% in first three days colostrum and milk
Lactose% Days Figure 3.5(b) Average Lactose% in first three days colostrum and milk

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3.7: Estimation of Total Solid:

Table 3.7 is showing the total solid content. The sample D1 (first day colostrum) has the total solid contents 26.25%, 26.26%, and 26.22%. Average total solid in the sample D1 was 26.26%. The sample D2 (second day colostrum) has the total solids i.e. 18.86%, 18.89%, and 18.87%, average of which was 18.87%. The sample D3 has the total solids 15.05%, 15.03% and 15.04%. Average total solid in the sample D3 was 15.04%. Sample D showed 11.81%, 11.83%, and 11.87% of total solid contents. Average total solid of sample D was 11.84%. Fig.3.7 (a) is showing the results of total solids of different samples D1, D2, D3, and D. The Fig.3.7 (b) is showing the average values of the total solids from sample D1 to sample D3, and D (mature milk) of the goat. It showed that there took place a continuous decrease in the total solid contents from sample D1 to sample D3, and sample D.
Table.3.7. Total solids in first three days colostrum and milk

Days after parturitions Total solids in first three days colostrum and milk Average %
1 D1 D2 D3 D 26.25 18.86 15.05 11.81
2 26.26 18.89 15.03 11.83
3 26.22 18.87 15.04 11.87
26.26 18.87 15.04 11.84

42
Total solids Readings Figure 3.7(a): Total solids in first three days colostrum and milk
Total solids% Days Figure 3.7(b) Average Total solids% in first three days colostrum and milk

43
Conclusions
The current study revealed that there took place a lot of variations within the constituents of the colostrum from first day to third day, and milk showed a widest range of deviations from the first day colostrum in many respects. It was analyzed that fat, proteins, ash, and total solids were much higher in the first day colostrum i.e. 7.8%, 14.17%, 1.05%, and 26.26% respectively. On third day fat, proteins, ash, and total solids were decreased to 4.8%, 4.52%, 0.87%, and 15.04% respectively. However the lactose and moisture were lower in the first day colostrum i.e.2.93% and 73.74% respectively. Lactose and moisture latter on increased to 4.43% and 84.96% respectively. The concentrations of the major colostrum constituents changed significantly after the birth, the level of different constituents on the third day was very similar to those of the mature milk.

44
CHAPTER NO.4 Discussion

45
4. Discussion
Colostrum is the nutrient rich pre-milk that contains substances which protect the infant from infections and other diseases and helps in the proper growth of the neonate (Ogra.,1983). It is rich in proteins, fats, minerals, and total solids (Siber, 1992). Colostrum differs from normal milk in many ways. It has high contents of total solids, fats, proteins, vitamins, Ig and is low in lactose (Corbett, 1997) and water contents. The first three days colostrum of the goat samples were analyzed for the determination of proteins, fats, ash, moisture, acidity and lactose. Colostrum was analyzed to compare its contents with mature milk. 4.1. The moisture contents In this study moisture of colostrum samples was determined by the Gravimetric (oven drying) method. The moisture content of the first day colostrum was 73.74%, second day colostrum was 81.13% and third day colostrum moisture was 84.96%.Similar values were reported by other workers (Omneis, 1904; Frahm, 1926; Williams et al., 1976; Puente et al., 1994; Tener, 1956; and Jenness, 1979). Siegfeld (1906) reported 71.84% and 84.46% moisture in first & third day colostrum respectively. Average data of moisture content of third day colostrum of fourteen goats showed 85.42% moisture (Burgman & Turner, 1936). The results of other worker were very close to our findings (Jenness, 1980; Devendra, 1972; Dozet, 1973; Ming et al.,2001; Salama et al.,2007; Jenness, 1979; Mba et al.,1975) Colostrum contains less moisture as compared to the mature milk. This is probably due to the sticky nature of colostrum. First day colostrum contained high amount of proteins particularly IgG (Georgiev, 2005) and high amount of fats and other solid contents. All solid contents maintain a low moisture level in the first day
colostrum, then a sudden decrease occurred in the solid contents particularly proteins, fats, etc, which resulted in a sharp increase of moisture level.

46
4.2. The Protein contents In our present studies proteins were estimated by using the Kjehldahl method for 3 days colostrum, and mature milk, which were 14.17%, 7.12%, 4.52%, and 3.44% respectively. There was a decrease in the protein content after the first day colostrum. The protein content variations in the compositional constituents of the goat milk and colostrum are influenced by many factors, such as stage of lactation, diet, breed, age, health, season, environment, etc (Schmidt, 1971). Siegfeld (1908) estimated 20.62% & 4.46% protein in first & third day colostrum respectively. Puente et al (1994) calculated 10.3+3.7%, 6.8+1.6% protein in first & second day colostrum respectively. Bergman and Turner (1936) presented average data from the different researcher in reference to goat colostral composition, showed 9.13% & 4.27% protein in first & third day colostrum respectively. Williams et al (1976) also showed similar results in the second day colostrum. Ranawana and Kellaway (1977) calculated 3.39%, Maes et al (1976) calculated 3.52%, Jensen (1995) calculated 3.1% protein content in the mature milk of the goats. There was a decrease in protein content from first to third day colostrum and mature milk. This could be attributed to the sharp fall of the concentrations of the immunoglobulins fractions, especially IgG (Georgiev, 2005). The immunoglobulins content in the first day colostrum is always very high in proportions and account for nearly about 50% of the total protein concentrations (IIiev, 1988; Levieux, 1999; Rauprich et al., 2000; Blum and Hammon, 2000; Tomov, 2002; Blum and Baumrucker, 2002). The first day milking contains a large amount of IgG which is given to the newborn, and promote neonatal host defense during the early days of life (Tomov, 1989; Blum and Baumrucker, 2002). Decrease in total milk protein concentration in the mature milk is probably due to dilution of the colostrum resulting from the increased milk production (Ontsouka et al., 2003).

47
4.3. The Fat contents Fats contents were analyzed by Gerbers Method in our studies. Fat content of the first day was 7.8%, on second day 6.53%, on third day 4.8%, and in mature milk it was 3.5%. Bergman and Turner (1936) estimated 8.21% fat in the first day, 5.15% in second day, 4.64% in third day colostrum respectively. Puente et al (1994) estimated 8.5+2.6% and 8.2+2.1% fats in first and second day colostrum. Frahm (1926) estimated 6.58% fats on second day, 5.36% on third day after parturition. Ali and Hassan (1990) reported the fat content with a mean of 3.61% in goat milk; in another other study by Ming et al (2001) fat in goat milk was 3.63%. Jensen (1995) reported 3.5% fats in the goat milk. Milk fat results of our studies are much consistent with many other workers like Mba et al (1975), Devendra (1972), Graf et al (1970), and Uusi-Rauva et al (1970). There are somewhat variations in the results as compared to other studies; these variations in fat content arose because of the difference in fat estimation procedures. These variations also arose due to the difference in concentrations of the fatsoluble vitamins among individuals as well as the maternal reserve status, diet, and season (Kehoe et al., 2007). Regional climate of the mammal also influences the colostrum and milk compositions (Brendehaug and Abrahamsen, 1986; Haenlein, 1996).
4.4. The Lactose contents The lactose content during this study was 2.93% in first day colostrum, 3.87%, and 4.43% in second & third day respectively. Where as the mature milk contained 4.8% lactose. The results of our studies are consistent with the results worked out by Steinegger (1898), Omneis (1904), Siegfeld (1906), and with the average data provided by Bergman and Turner (1936) of first three days colostrum samples. Puente et al (1994) estimated 4.7+0.6% lactose in goat milk. Ming et al (2001) estimated 4.47 + 0.15% lactose in the milk of Commingled goat. According to Jensen (1995) goat contains 4.6% lactose in the mature milk. Ranawana and Kellaway (1977) showed 4.85%, Sachdeva et al (1974) estimated 4.72%, Mba et al (1975) estimated 4.54% and

48
4.72%, and Uusi-Rauva et al (1970) estimated 4.48% lactose in the mature milk. Lactose concentrations are reduced in colostrum and act inversely of other constituents, such as solids, protein and ash, which are all found in high concentrations and decrease over time (Kehoe et al., 2007). According to Kuhn (1983), it is clear that low level of plasma glucose and colostral lactalbumin is the main cause of the low level of lactose in the colostrum. Lactose production causes water influx in milk through osmotic effects, and values were lower in colostrum than in mature milk. However, concentrations of Na and Cl, which are, osmotically active molecules in the milk, were elevated in colostrum (so wise ash contents are high) as compared with mature milk (Ontsouka et al., 2003). The electrolyte transfer from blood into milk through leaky tight junctions (Nguyen and Neville, 1998) are likely to be expected for increase in the milk volume during the colostral period despite relatively low lactose secretion (Ontsouka et al., 2003). 4.5. The Ash content During this study the ash contents were 1.05%, 0.96, and 0.87% for first, second, and third day colostrum respectively, the ash content of mature milk was similar with 3rd day colostrum i.e.0.87%. Steinegger (1898) calculated 1.00%, Omneis (1904) 1.15%, Siegfeld (1908) 1.27%, and Bergman and Turner (1936) 1.04% ash contents in the first day colostrum. The ash contents calculated in our studies also deviated from some earlier studies and that could be due to change in climate, diets, breed, testing techniques and much other dissimilarity which may exist. However, the ash content of the mature milk showed much consistency. Dozet (1973) from the Yugoslavia estimated 0.88% ash. Kotschopoulos (1940), Williams et al (1976), Uusi-Rauva et al (1970), Sachdeva et al (1974), and Akinsoyinu et al (1977) showed the same results of the ash contents in the mature milk of goat. The decrease in the ash contents is probably due to the low level of lactose in colostrum, because lactose and ash contents maintain the osmolarity of the milk and colostrum. With the low level of lactose to maintain the osmolarity to the normal values, it is necessary to have high level of ash contents. Therefore the ash contents in the first

49
day colostrum were high and lactose contents were low, as the lactose percentage in the proceeding days increased then a gradual decrease was observed in the ash contents. 4.6. Total solid contents Our results revealed 26.26% total solids in the first day colostrum, 18.87% in second day, 15.04% in the third day colostrum, and finally 11.84% in the mature milk of the goat. Total solids were much higher in the first day colostrum and on next days the solids content went on decreasing in the mature milk. Puente et al (1994) estimated 24.0+5.7% total solids in first day, 20.2+3.2% in second day colostrum, and 13.6+1.3% total solid in mature milk after 30 days of lambing in goat. Scheurlen (1908) estimated 28.70% total solids in the first day colostrum. Frahm (1926) observed 24.42% in first day, 16.48% and 15.45% total solids in second and third day colostrum respectively. Bergman and Turner (1936) showed, 24.92%, 16.10%, and 14.58% total solids in first, second, and third day colostrum respectively after the parturition time. These slight variations in the values of different studies resulted because of many factors such as, diets, breed, age, health, season, and environment, etc (Schmidt, 1971). Ming et al (2001) estimated 12.38+0.71% with the range of 11.17-13.44% of total solids in the mature milk. Parkash and Jenness (1968) calculated 13.3% and Jensen (1995) gave 12% total solids in mature milk of goat. Many other workers like, Ranawana and Kellaway (1977), Uusi-Rauva et al (1970), Graf et al (1970), Mba et al (1975), Devendra (1972), and Dozet (1973) estimated the results which are very close to our studies. The reason for high total solids content in the first day colostrum was its high contents of proteins particularly IgG contents (Guidry et al., 1980b; Butler, 1983; Larson, 1992; Vacher and Blum, 1993). Concentrations of other protein fractions such as lactoglobulin, lactoferrin and transferrins are known to be also higher in colostrum than in mature milk (Sanchez et al., 1988; Ye-Xiuyn and Yoshida, 1995). High fatty acids contents and ash content also contributed to this high level of total solids in the first day colostrum.

50
CHAPTER NO.5 REFERENCES

51
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Comparative Analysis of Goat Milk and Colostrum

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Comparative Analysis of Goat Milk and Colostrum

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A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.

SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

SARWAR ALLAH DITTA.

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

Dedicated To My Mother The Most Perfect Lady Of The World

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

1.1 Composition of the colostrum

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A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

Sphingomylein (Patton and Keenan, 1971). Cerebrosides, glucosyl-

A THESIS BY MR.SARWAR A.D. ZOOLOGY DPTT.SCIENCE COLLEGE WAHDAT

1.1.4. Minerals and Salts:

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1.2. Significances of the colostrum

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1.4. Mechanism of immunity:

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1.5. Objectives of the study

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CHAPTER NO.2 MATERIALS AND METHODS

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2. MATERIALS AND METHODS

2.2. List of Apparatus

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2.3. List of Reagents & Solutions

2.4. Principles of tests

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2.5. Determination of Protein Content (Kjeldhal Method)

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By applying the following formula we can calculate the protein %.

= (B) mg N Weight of sample

Protein % age = (C) x Dairy factor (6.38)

2.6. Determination of Moisture Contents (Oven Drying Method)

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2.7. Determination of Ash Contents

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2.8. Determination of Fat Contents (Butyrometer Method)

2.9. Determination of Lactose in Colostrum and milk of the goat

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CHAPTER NO.3 RESULTS

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Table 3.1 Days after parturitions D1 D2 D3 D

Protein % in first three days colostrum and milk

1 14.18 7.12 4.53 3.45

2 14.15 7.14 4.50 3.43

3 14.17 7.11 4.53 3.44

Average % 14.17 7.12 4.52 3.44

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3.2: Estimation of Fat contents: