www.wjpps.com Vol 6, Issue 12, 2017.

888

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

COMPARATIVE STUDIES ON DIFFERENT SOURCES OF MILK:

OUR LABORATORY EXPERIENCES

*Dr. Amit Kumar Dutta, Ph.D

Professor & Head, School of Biological and Chemical Sciences, MATS University,

Raipur(CG)- 492004.

ABSTRACT

Milk has been a food source for humans since prehistoric times; from

human, goat, buffalo, sheep, yak, to the focus of this section -

domesticated cow milk (genus Bos). Milk and honey are the only

articles of diet whose sole function in nature is food. It is not

surprising, therefore, that the nutritional value of milk is high. Milk

from cows, sheep, goats and humans is rich in microorganisms. Lactic

acid bacteria, the most abundant microorganisms found in milk,

facilitate dairy fermentation and promote health. Milk samples were

collected from the different sources and the different areas of Raipur

district. We have collected basically 3 types of Milk, like Packet Milk

which is manufactured from one Certified Well Established Dairy Company. Second one, we

kept as Buffalo Milk for our experiment and in the last we are taken Cow Milk for our

research experiment. During the time of isolation of bacteria, we were observing

morphological as well as Biochemical characters. In the morphology parts, we studied colony

characters and cell morphology. The acid can build up and causes the protein in curdle.

Notably of the genera Bacillus, Micrococcus and Proteus, digest the casein in milk. The

casein then curdles out of the milk. Hence from our all above experiment from all the three

different samples conclude that Cow and Buffalos Milk are better than Packet Milk whether it

may be local or certified company. One of the most direct ways of studying the flora of milk

is to set milk samples under various environmental conditions and determine which

populations emerge and their sequence of emergence.

KEYWORDS: Samples, Serial Dilution, Culture, Morphological Examination, Biochemical

Analysis, Staining.

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 6.647

Volume 6, Issue 12, 888-901 Research Article ISSN 2278 – 4357

Article Received on

03 October 2017,

Revised on 24 October 2017,

Accepted on 13 Nov. 2017

DOI: 10.20959/wjpps201712-10559

*Corresponding Author

Dr. Amit Kumar Dutta

Professor & Head, School of

Biological and Chemical

Sciences, MATS University,

Raipur(CG)- 492004.

www.wjpps.com Vol 6, Issue 12, 2017.

889

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

INTRODUCTION

Milk is a pale liquid produced by the mammary glands of mammals. It is the primary source

of nutrition for infant mammals (including humans who breastfeed) before they are able

to digest other types of food. Early-lactation milk contains colostrum, which carries the

mother's antibodies to its young and can reduce the risk of many diseases. It contains many

other nutrients including protein and lactose. Throughout the world, there are more than six

billion consumers of milk and milk products. Over 750 million people live in dairy farming

households.[1,24]

Figure Showing the Milk Samples for the Experiments

Physical properties

Density

Density is defined as an object’s mass divide by it’s volume. It depends on the temperature of

the object, composition of the material, and whether or not the object contains air. The

density of milk products can be used to convert volume into mass and mass into volume, to

estimate the amount of solids present in milk, and to calculate other physical properties. The

density of cow’s milk usually varies between 1.028 and 1.038 g/cm3

.

Appearance

The opacity of milk is due to its content of suspended particles of fat, protein, and minerals.

The colour varies from white to yellow depending on the carotene content of the fat. Skim

milk is more transparent and has a slightly bluish colour.

Freezing Point

The freezing point of milk is lower than the freezing point of water because of the dissolved

components in milk. Measuring the freezing point is used as a legal standard to determine if

milk has been diluted with water. The freezing point point of milk is -0.5520C or 31

0F.

www.wjpps.com Vol 6, Issue 12, 2017.

890

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

Chemical properties

pH

The pH of milk is higher, or more alkaline, outside of the cow than inside the cow due to loss

of carbon dioxide to the air. The ph of milk is never determined immediately after milking

because the processing milk goes through removes dissolved gasses. The pH is determined

after processing the milk to assure that lactic acid is being produced at the desired rate by

added microorganisms during the preparation of cheeses and fermented milk. The casein in

milk forms into a curd or a gel at a pH of 4.6.[1,2]

Titratable Acidity

Titratable acidity is the amount of alkali required to bring the pH to neutrality. This property

of milk is used to determine bacterial growth during fermentation, such as cheese and yogurt

making, as well as compliance with cleanliness standards. Naturally, there is no lactic acid in

fresh bovine milk, however, lactic acid can be produced by bacterial contamination, but this

is uncommon. The titratable acidity is due to the casein and phosphates.[2,25]

The role of milk in nature is to nourish and provide immunological protection for the

mammalian young. Milk has been a food source for humans since prehistoric times; from

human, goat, buffalo, sheep, yak, to the focus of this section - domesticated cow milk

(genus Bos). Milk and honey are the only articles of diet whose sole function in nature is

food. It is not surprising, therefore, that the nutritional value of milk is high. Milk is also a

very complex food with over 100,000 different molecular species found. There are many

factors that can affect milk composition such as breed variations, cow to cow variations, herd

to herd variations - including management and feed considerations, seasonal variations, and

geographic variations. With all this in mind, only an approximate composition of milk can be

given:

The following terms are used to describe milk fractions:

Plasma = milk - fat (skim milk):- what is left after you separate the fat globules, equivalent

to skim milk for practical purpose.

Serum = plasma - casein micelles (whey):- what is left after you take away both fat globules

and casein micelles, equivalent to cheese whey for most practical purpose.

Milk permeate= what is left after you take away fat globules, casein micelles, and whey

protein.

Solids-not-fat (SNF) = proteins, lactose, minerals, acids, enzymes, vitamins.

www.wjpps.com Vol 6, Issue 12, 2017.

891

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

Total Milk Solids = fat + SNF

Composition of milk

Constituents Composition Proportion Solids (%)

Fat 37.0 28.9

Protein casein 27.6 21.6

Protein whey 6.4 5.0

Non-protein nitrogen 1.9 1.5

Lactose 48.0 37.5

Ash 7.0 5.5

Total solids 127.9 100.0

This chart shows the composition of milk, detailing the major components as proportion

of solids.

The main source of lipid is from the cow’s food. Little synthesis of fatty acids occurs in the

mammary gland. When the cow consumes food, the lipid is hydrolyzed to free fatty acids

within the rumen of the cow. This means that unsaturated fatty acid is usually hydrogenated

into saturated fatty acids. Hydrogenation is the addition of hydrogen on unsaturated bonds

between carbon atoms.[3-6]

Structure of Milk

www.wjpps.com Vol 6, Issue 12, 2017.

892

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

Looking at milk under a microscope, at low magnification (5X) a uniform but turbid liquid is

observed. At 500X magnification, spherical droplets of fat, known as fat globules, can be

seen. At even higher magnification (50,000X), the casein micelles can be observed. The main

structural components of milk, fat globules and casein micelles, will be examined in more

detail later.

Bacteria present in milk

Milk from cows, sheep, goats and humans is rich in microorganisms. Lactic acid bacteria, the

most abundant microorganisms found in milk, facilitate dairy fermentation and promote

health. Other microorganisms in milk cause spoilage and may lead to illness. Pasteurization

kills most or all of these bacteria, so commercially processed milk contains few beneficial

bacteria. Fermented milk products and raw milk are abundant in healthy bacteria. However,

the Centres for Disease Control and Prevention strongly advises against consuming raw milk,

as it may contain other harmful bacteria that can make you sick or kill you.

Lactobacillus

Lactobacillus is a species of lactic acid bacteria that naturally populate healthy intestines.

Lactobacillus casei and rhamnosus are abundant in raw milk and are commonly used as

probiotics, or healthy bacteria. In a 2002 review of studies on the health benefits of lactic acid

bacteria, Aijaz Soomro and colleagues state that they help reduce lactose intolerance,

alleviate diarrhea, lower blood cholesterol, increase certain immune responses and may help

in the prevention of cancer. They may also aid in digestion, suppress the growth of pathogens

in the intestines, reduce toxicity of intestinal contents and improve intestinal function.

Streptococcus

Another dominant species of lactic acid bacteria found in raw cow milk is streptococcus.

These bacteria are frequently used for culturing cheese and yogurt as they ferment lactose,

the primary sugar in milk, to lactate. They also produce lactase, the enzyme that digests

lactose, helping people with lactose intolerance digest milk more efficiently. Streptococcus

thermophilus is a probiotic that helps improve digestion, enhances immunity, helps fight the

spread of harmful bacteria in the gut and confers a number of other health benefits.

Bifidobacterium

Bifidobacterium is a species of lactic acid bacteria found in human milk and fermented dairy

products. In breastfed babies, these bacteria contribute to infant digestion by aiding in the

www.wjpps.com Vol 6, Issue 12, 2017.

893

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

breakdown of complex foods such as proteins and sugars. In a 2012 review in "FEMS

Microbiology Reviews," Lisa Quigley and colleagues state that various bifidobacteria have

positive effects on health, including protection against infection by pathogenic bacteria,

stimulation of the immune system, decrease of cancer risk, lowering of serum cholesterol and

aiding in the digestion of lactose for those who are lactose intolerant.

Enterococcus

Enterococcus is a species of bacteria found in abundance in raw cow, goat, sheep and human

milk. Enterococcus faecalis is a normal inhabitant of the gut flora of both humans and

animals. In a 2003 study in the "Journal of Nutrition," Jalil Benyacoub and colleagues note

that these lactic acid bacteria, which protect against infection and help to relieve diarrhea, are

particularly beneficial for animals as well as humans and are frequently added as probiotics to

dog foods.[1,7-8,21-23]

Application of milk

1. Casein

The most important applications of caseins in fabricated foods are cheese analogues,

synthetic whipping creams, cream liqueurs, fabricated meats, some cereal products, various

dietetic foods and as an emulsifier in coffee whiteners. Schematic representation of casein

submicelles and casein micelle composed of submicelles held together by calcium phosphate.

2. Whey Protein

Physical, chemical and structural properties of whey proteins determine their functional

properties. As the primary food application of whey proteins is as emulsifiers, the optimum

conditions under which they act have been extensively studied. Some influential factors are

processing conditions, the method of isolation, environmental conditions (e.g., pH,

temperature, ionic strength, etc.), and interaction with other food components. Furthermore,

its ability to form gels capable of holding water, lipids, and other components while

providing textural properties makes them perfect to be used in processed meat, dairy and

bakery products. Regarding its foaming properties, they mainly depend on the degree of the

protein denaturation.[9-13]

www.wjpps.com Vol 6, Issue 12, 2017.

894

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

Figure Showing the Products prepared from Milk

MATERIALS AND METHODS

Collection Of Milk Sample

Milk samples were collected from the different sources and the different areas of Raipur

district. We have collected basically 3 types of Milk, like Packet Milk which is manufactured

from one Certified Well Established Dairy Company. Second one, we kept as Buffalo Milk

for our experiment and in the last we are taken Cow Milk for our research experiment.

Figure Showing the Milk Samples in the LAF for the Experiments

Isolation Of Bacteria From Milk

After collection, the freshly milk samples were cultured for the observation of bacteria which

are present in the different milk samples, if any, to see the quality of milk among the different

samples. Bacteria were isolated from milk sample through serial dilution method on Nutrient

Agar Medium(NAM) plates. Prepare 10 test tubes, one with 10 ml and rest of test tubes with

9 ml of distilled water. Label all tube. So don’t get confused once begin with the dilution.

Adding 1gm of milk sample to first tube and serial dilution in to the following tubes and then

thoroughly mix your solution before starting and dilution. The 1st tube will be 1:10 followed

by 2nd

tube will be 1:100, 3rd

tube will be 1: 1000 and 10th

tube will be 1: 10000000000. After

that draw 1ml of undiluted solution from test tube with a pipette and transfer it to the test

www.wjpps.com Vol 6, Issue 12, 2017.

895

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

labelled 10-1

containing 9ml of the dilution liquid and mix thoroughly. For the second serial

dilution take 1ml of solution from tube 10-1

and add it to the 10ml of dilution liquid in the

tube 10-2

following dilution. Extend this procedure to perform longer serial dilution. This

process may be repeated as many times necessary to achieve the desired solution. In an

experiment involving concentration curves, dilution to create a series of solution with dilution

of 10-1

, 10-2

......10-10

. Then select 10-4

, 10-5

and 10-6

dilution. 100 of dilution selected

sample were taken and pour into pre-poured NAM plate and spread through spreader. The

plate was incubated at 370C for 24 hour for proper growth of bacteria. After proper growth

each different colony of bacteria should pure cultured into NAM slants and store for further

analysis.

Characterization Of Bacteria Isolated From Milk

During the time of isolation of bacteria, we were observing morphological as well as

Biochemical characters. In the morphology parts, we studied colony characters and cell

morphology.

Gram’s staining was performed for the identification of Gram positive and Gram negative

bacteria. Apart from this, we are also performed IMViC tests followed by (i) Indole

production, (ii) Methyl- Blue. Tryptophan is an essential amino acid, is oxidized by some

bacteria by the enzyme tryptophanase resulting in the formation of indole, pyruvic acid and

ammonia. The indole production test was confirmed by Kovac’s reagent

(dimethylaminobenzaldehyde) which produces a cherry-red reagent layer shows positive

result of indole production. Methyl Blue and Voges-Proskauer test (MRVP) tests are used

to differentiate two major types of facultative anaerobic enteric bacteria that produces large

amounts of acid and those that produces the neutral product, acetoin as end product. In MR

test the colour remained red if pH ranges in 4 shows positive result while turning of methyl

red to yellow is negative test. In VP test the development of crimson-to ruby pink (red)

colour is indication of positive test of VP while no change in colour is shows negative

test.[7,10,18-20]

www.wjpps.com Vol 6, Issue 12, 2017.

896

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

RESULTS AND OBSERVATIONS

Figure Showing the Biochemical Test for checking the Grading & Quality of Milk.

Figure Showing the Culture of Milk in Series of Dilution for the Growth of Bacteria

Sample:- Packet Milk.

Figure Showing the Culture of Milk in Series of Dilution for the Growth of Bacteria

Sample:- Buffalo Milk.

Figure Showing Bacterial Colonies found in Milk in all Three Samples.

www.wjpps.com Vol 6, Issue 12, 2017.

897

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

Figure Showing the Different Types of Bacteria found in Milk in all Three Samples.

DISCUSSION AND CONCLUSION

Milk is used to make yogurt, cheese, ice milk, pudding, hot chocolate and french toast. Milk

is often added to dry breakfast cereal, porridge and granola. Milk is often served in coffee and

tea. Steamed milk is used to prepare espresso-based drinks such as cafe latte. Besides serving

as a beverage or source of food, milk has been described as used by farmers and gardeners as

an organic fungicide and fertilizer, however, its effectiveness is debated. Diluted milk

solutions have been demonstrated to provide an effective method of preventing powdery

mildew on grape vines, while showing it is unlikely to harm the plant.

The numbered list below identifies seven types of bacteria according to how they change the

properties of milk. Often these changes are negative (spoilage) but as we already found that

many of these bacteria are important to the development of cheese flavour. Psychrotrophic

refers to microorganisms which are able to grow at temperatures less than 70C. Cold milk

storage and transport selects for psychrotrophic bacteria which are often proteolytic and

lipolytic. Common psychrotrophic bacteria in milk are species of Micrococci, Bacilli,

Staphyloccoci, Lactobacilli, Pseudomonas and Coliforms. Pseudomonas species are the most

common and typically have the most impact on quality. At temperatures of 2- 40C, bacterial

growth in milk is mainly due to strains of Pseudomonas flourescens. Little growth occurs at

temperature less than 20C. Spore forming bacteria are able to exist in a highly stable form

called 'spores'. In the spore state, these bacteria are able to withstand greater extremes of

acidity, temperature and desiccation. Enzymes are biological catalysts that accelerate the

rates of biochemical reactions. Bacterial enzymes are most significant to milk spoilage and

cheese ripening but it is important to distinguish between the enzyme and the bacterial

source. Psychrotrophic bacteria produce heat stable enzymes which remain active in milk and

cheese even after the bacteria are killed by pasteurization.[14-17,19,21]

www.wjpps.com Vol 6, Issue 12, 2017.

898

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

Keeping the above experiment and hands on research in mind, microorganisms play an

important role for the impact on milk quality. Lactic Acid Bacteria which ferment lactose to

lactic acid and other end products. LAB is able to readily metabolize lactose so they have

some competitive advantage over other microorganisms. Proteolytic bacteria which degrade

protein and cause bitterness and putrefaction. Most important in cheese milk are species of

Pseudomonas which are psychrotrophic and produce heat stable lipases. Bacillus which form

heat stable spores and survive pasteurization. Lipolytic bacteria which degrade fats and

produce lipolytic rancidity. Several psychrotrophic species of Pseudomonas produce heat

stable lipases as well as proteases. Gas producing microorganisms which cause cheese

openness, floating curd in cottage cheese and gassy milk. Yeasts are always present in milk

and are common contaminants during the cheese making process. Some lactic cultures, called

hetero fermentative, also produce carbon dioxide. Ropy bacteria cause stringy milk due to

excretion of gummy polysaccharides. Usually ropy bacteria such as Alcaligenes

viscolactis are undesirable. Sweet curdling bacteria produce rennet-like enzymes which may

coagulate milk. The importance of milk in human culture is attested to by the numerous

expressions embedded in our languages. In ancient Greek mythology, the goddess Hera

spilled her breast milk after refusing to feed Heracles, resulting in the Milky Way in the sky.

In many countries, butter is traditionally made from fermented milk rather than cream. It can

take several hours of churning to produce workable butter grains from fermented milk. The

natural flora of milk is plentiful and diverse. Various types of bacteria emerge in a milk

sample when it is placed at different temperatures. Certain bacterial species such as

Lactobacillus and Streptococcus produce lactic acid from the lactose and make the milk sour.

The acid can build up and causes the protein in curdle. Notably of the genera Bacillus,

Micrococcus and Proteus, digest the casein in milk. The casein then curdles out of the milk.

Bacteria also attack the milk’s butterfat. Hence from our all above experiment from all the

three different samples conclude that Cow and Buffalos Milk are better than Packet Milk

whether it may be local or certified company.

One of the most direct ways of studying the flora of milk is to set milk samples under various

environmental conditions and determine which populations emerge and their sequence of

emergence.

www.wjpps.com Vol 6, Issue 12, 2017.

899

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

ACKNOWLEDGEMENT

We are really thankful to the Hon’ble Vice-Chancellor and Respected Registrar for providing

us to carry out this research at School of Biological & Chemical Sciences, MATS University,

Raipur(CG). Our sincere thanks towards our parents for supporting us with directly and

indirectly in different ways for this review based microbiological as well as biotechnological

research.

REFERENCES

1. Uruakpa, F. O.; Ismond, M. A. H.; Akobundu, E. N. T. (2002). "Colostrum and its

benefits: A review". Nutrition Research. 22(6): 755–767.

2. Blood DC, Studdert VP, Gay CC (2007). Saunders Comprehensive Veterinary

Dictionary. St. Louis, Missouri, USA: Saunders Elsevier.

3. The World Health Organization's infant feeding recommendation WHO, based on

"Global strategy on infant and young child feeding" (2002). Retrieved February 8: 2013.

4. Dettwyler, Katherine A. (October 1997). "When to Wean". Natural History. Retrieved

February 8: 2013.

5. Basnet, S.; Schneider, M.; Gazit, A.; Mander, G.; Doctor, A. (April 2010). "Fresh Goat's

Milk for Infants: Myths and Realities—A Review". Pediatrics. 125(4): e973–977.

6. Curry, Andrew (July 31, 2013). "Archaeology: The milk revolution". Nature. 500(7460):

pp.20–22.

7. McGee, Harold (2004) [1984]. "Milk and Dairy Products". On Food and Cooking: The

Science and Lore of the Kitchen (2nd ed.). New York: Scribner. pp. 7–67.

8. "World's No 1 Milk Producer". Indiadairy.com. Retrieved August 28, 2010.

9. Goff, Douglas. "Introduction to Dairy Science and Technology: Milk History,

Consumption, Production, and Composition: World-wide Milk Consumption and

Production". Dairy Science and Technology. University of Guelph. Retrieved November

12, 2014.

10. Codex Alimentarius Commission. "General Standard for the Use of Dairy Terms 206-

1999" (PDF).

11. Gussekloo, S.W.S. (2006). "Chapter 2: Feeding Structures in Birds". In Bels, V. Feeding

in Domestic Vertebrates: From Structure to Behaviour. CABI Publishing. pp. 22.

12. Oftedal, Olav T. (2002). "The mammary gland and its origin during synapsid evolution".

Journal of Mammary Gland Biology and Neoplasia. 7(3): 225–252.

www.wjpps.com Vol 6, Issue 12, 2017.

900

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

13. Oftedal, Olav T. (2002). "The origin of lactation as a water source for parchment-shelled

eggs". Journal of Mammary Gland Biology and Neoplasia. 7 3): 253–66.

14. "Lactating on Eggs". Nationalzoo.si.edu. July 14, 2003. Archived from the original on

April 14, 2009. Retrieved March 8, 2009.

15. Lefevre CM, Sharp JA, Nicholas KR (2010). "Evolution of lactation: ancient origin and

extreme adaptations of the lactation system". Annual Review of Genomics and Human

Genetics. 11(1): 219–238.

16. Vorbach C, Capecchi MR, Penninger JM (2006). "Evolution of the mammary gland from

the innate immune system?". BioEssays. 28(6): 606–616.

17. Goldman A.S. (2002). "Evolution of the mammary gland defense system and the

ontogeny of the immune system" (PDF). Journal of Mammary Gland Biology and

Neoplasia. 7(3): 277–289.

18. Hu, Yaoming; Meng, Jin; Clark, James M. "A New Tritylodontid from the Upper Jurassic

of Xinjiang, China". Acta Palaeontologica Polonica. 54(3): 385–391.

19. Bellwood, Peter (2005). "The Beginnings of Agriculture in Southwest Asia". First

Farmers: the origins of agricultural societies. Malden, MA: Blackwell Publishing. 44–68.

20. Bellwood, Peter (2005). "Early Agriculture in the Americas". First Farmers: the origins of

agricultural societies. Malden, MA: Blackwell Publishing. 146–179.

21. Beja-Pereira, A.; Caramelli, D.; Lalueza-Fox, C.; Vernesi, C.; Ferrand, N.; Casoli, A.;

Goyache, F.; Royo, L. J.; Conti, S.; Lari, M.; Martini, A.; Ouragh, L.; Magid, A.; Atash,

A.; Zsolnai, A.; Boscato, P.; Triantaphylidis, C.; Ploumi, K.; Sineo, L.; Mallegni, F.;

Taberlet, P.; Erhardt, G.; Sampietro, L.; Bertranpetit, J.; Barbujani, G.; Luikart, G.;

Bertorelle, G. (2006). "The origin of European cattle: Evidence from modern and ancient

DNA". Proceedings of the National Academy of Sciences. 103(21): 8113–8118.

22. Sherratt, Andrew (1981). "Plough and pastoralism: aspects of the secondary products

revolution". In Hodder, I.; Isaac, G.; Hammond, N. Pattern of the Past: Studies in honour

of David Clarke. Cambridge: Cambridge University Press. 261–305.

23. Vigne, D.; Helmer, J.-D. (2007). "Was milk a 'secondary product' in the Old World

Neolithisation process? Its role in the domestication of cattle, sheep and goats" (PDF).

Anthropozoologica. 42(2): 9–40.

24. Evershed, R. P.; Payne, S.; Sherratt, A. G.; Copley, M. S.; Coolidge, J.; Urem-Kotsu, D.;

Kotsakis, K.; Ozdoğan, M.; Ozdoğan, A. E.; Nieuwenhuyse, O.; Akkermans, P. M. M.

G.; Bailey, D.; Andeescu, R. R.; Campbell, S.; Farid, S.; Hodder, I.; Yalman, N.;

Ozbaşaran, M.; Biçakci, E.; Garfinkel, Y.; Levy, T.; Burton, M. M. (2008). "Earliest date

www.wjpps.com Vol 6, Issue 12, 2017.

901

Dutta. World Journal of Pharmacy and Pharmaceutical Sciences

for milk use in the Near East and southeastern Europe linked to cattle herding". Nature.

455(7212): 528–531.

25. Price, T. D. (2000). "Europe's first farmers: an introduction". In T. D. Price. Europe's

First Farmers. Cambridge: Cambridge University Press. 1–18.