International Journal of Diary Science Research. Assessment of fresh bulk milk Quality Available for Human Consumption in Quetta, City by leukocytes.

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Available online at http://www.urpjournals.com International Journal of Diary Science Research Universal Research Publications.

1 Department of Chemistry University of Balochistan, Pakistan. PCSIR Laboratories,. (P.C 87300). *Corresponding Author Email.Aziz1sh@Hotmail.com, Tel +93318077454.

1 Available online at International Journal of Diary Science Research Universal Research Publications. All rights reserved Assessment of fresh bulk milk Quality Available for Human Consumption in, City by leukocytes. *Muhammad Aziz 1, Zaheer ud din, Saima Jadoon 1. 1 Department of Chemistry University of Balochistan, Pakistan. PCSIR Laboratories,. (P.C 87300). *Corresponding Author .Aziz1sh@Hotmail.com, Tel Received 1 February 013; accepted 0 March 013 Abstract To evaluate the quality of bulk milk supplied to the consumers of, city. Eighteen (18) milk samples were collected from three regions of city, i.e. Brewery road, Sariab road and Sabzal road. Six (6) samples from each area with a ratio of 3:3 i.e. three milk samples from dairy farms and three samples from milk sale s were collected for observation. Meaurment. To know quality of milk following parameters were noted Colour,Odour, Temperature, Water content, ph and Somatic cell count. Result It was observed that 44.44% of milk sample opaque, 33.33% had creamy and.% had chalky colours, respectively, 66.66% of the milk samples had sweet while 33.33% of the milk samples had cheesy odour. ph of the milk samples recorded was 44.44% in the range of 6.0 to 6., 33.33% in the range of 6.3 to 6.7 while.% was in the range of ¾. The leukocyte count was.% in the range of 78, , 77; 55.55% in the range of 1,0,840-1,99,010 and.% in the range of,44,00-,78,909. Conclusion It was concluded that milk colour, odour, ph, water content and somatic cell count might be used as an easy tool / indicator to detect normal or abnormal milk within a short period of time. 013 Universal Research Publications. All rights reserved Key words: Mammalian milk,, W.B.C. INTRODUCTION Milk and dairy products are essential for maintaining the health of the public. It is therefore essential to know the quality of milk in any location. The milk of animals was used as food since prehistoric times, so it is not possible to state precisely when men first became aware about milk. It is the liquid food secreted by the mammary gland [1] for the nutrition and well-being of mammalian infants and also for adult life. It is actually a nutritious food designed to produce rapid growth in young mammals and contains more solid material than many of other common foods. The legal definitions of milk vary from country to country. Milk of all species contains approximately the same nutrients, differing only in proportions. The milk contain protein, carbohydrate, fat, minerals, vitamins, enzymes and various trace element [3,4].Approximately there are more than 00 chemical components have been identified in milk. The nutritional status of a population can be determined in various ways, milk quality assessment is the best single measure to define the nutritional status. To know the concentration of fresh milk is a great interest in the fields of dairy production Processing companies. The key objective of study evaluate the quality of milk through its colour, odour, ph, water content and leukocytes cell counts in the locally available market fresh milk without processing. SOMATIC CELL count in milk is used as an indirect measure of the degree of mammary gland status, the number of cell increase during infection in gland [8]. In these ways we also indirect measure the health status of animals too, In few only measure inorganic contents in powder milk [5]. Location To know milk composition in any location is essential because it put a huge impact on physicochemical factors, which directly affect the survival of the place in these regard we conduct a study in, city the capital of Balochistan province in Pakistan. The total area of is 1,04 sq m approximately it is located at the west edge of Pakistan and located in high altitude i.e 5,60 ft above from sea level. district is consider the urban district of balochistan at N E coordinates. Demand for 1

2 fresh milk is high as compared to processed milk and to full fill the milk demand in city, milk is transported to from adjacent area MATERIALS AND METHODS. Collection Samples Milk samples was collected during 006 from the selected areas (i.e. Sariab road, Sabzal road, and Brewery road with their respective surrounding areas) of the city. Half of these samples were collected from the dairy farms directly while half of the samples were collected from the sale s of the same areas. So that we can compare the quality of both milk samples that are available to the consumers. Each of the milk sample was collected in a glass caped bottle and kept on dark place [7]. REAGENTS ph buffer, Saline solution, Methylene blue, % acetic acid, Distilled water Colour, Odour and Temperature of milk samples For the observation of colour, odour and temperature of the milk samples,the samples was poured in a 100 ml glass beaker and allowed to settle down. After some time the colour was observed through standard colour strip scheme and odour was feel through using the natural smelling sense while temperature was noted by using the thermometer. Determination of Hydrogen ion concentration (ph) of milk samples For the determination of Hydrogen ion concentration (ph) of milk sample, electric ph meter was used. After stirring the whole milk sample by a glass rod, a part (50ml) of this sample was taken in a 100 ml glass beaker and Combined Electrode of the ph meter was placed about 3 cm deep in it. The reading of ph meter was noted after 30 seconds of placing these electrodes in the sample. The reading was recorded form the ph meter screen; combined electrode was removed from the sample and dried. For accuracy, the procedure was repeated. Determination of water contents in milk samples. Result and discussion TABLE No.1 Milk sample collecting area and type S.No. AREA DATE OF SAMPLE COLLECTION To detect the water content present in the milk, lactometer was used. 100 ml of milk sample already stirred and warmed (upto temperature 39 0 C) was taken in a graduated cylinder and lactometer was dipped in this cylinder and the result was noted on the lactometer. i.e. if the reading shows 0, it indicates the pureness of milk sample (with 0% of water addition), if shows ¾ it indicates 75% water. By this way the water content of all the milk samples was observed. For accuracy of the results the procedure was repeated for every sample. Estimation and counting of leukocytes in milk samples. Acidic diluting fluid does not destroy WBCs. Stain added to diluting fluids makes nuclei visible in a counting chamber and easily examine under the microscope. For the estimation and counting of leukocytes, procedure given by Brar, et, al 000 was followed [6]. About 0.5 of milk sample was drawn from the whole sample with the help of pipette having white bead. At the time of drawing diluting fluid (% acetic acid and 1 drop of methylene blue) gently revolve the pipette. This keeps bead in motion and prevents air bubbles sticking to the bulb. During shaking the pipette for few seconds the thumb was placed on the tip of the pipette. Then remove rubber tubing and hold the pipette horizontally between thumb and finger. Allow the cells to settle down for 1 to minutes. Locate the upper left larger corner square (divided into 16 intermediate squares ) using low power objective. Adjust the light so that marking lines are clearly visible. The cells were count in 16 intermediate squares. To avoid counting of cells twice, only those cells were counted, which touch left and upper boundary lines. The counting was taken in all the four squares marked W making a total area of 4 sq. mm. The calculation was performed by using the following formula: Calculation. Number of leukocytes / cu. Mm = Leukocytes in 4 WBC squares x 50 x NAME OF DAIRY / MILK SHOP Allah bux Dairy Farm DAIRY Jan Muhammad Dairy Farm DAIRY Sahib Dad Dairy Farm DAIRY Ali Suleman Dairy Farm & bakers Rahal Milk Center Location name Wahdat colony, Brewery Road, Near Killa Sheikh Hussaini, Brewery road Pashtoon Bagh, Kharotabad, Sabzal road, Golimar chowk, brewery road, Near BMC main hostel, brewery road, Muhammadi Bakers & Milk Shop Joint road,

3 Sher Muhammad Dairy Farm DAIRY Dawat Dairy Farm DAIRY Mukhtiar Dairy Farm DAIRY Shahwani Milk Shop Riaz Bakers & Milk Shop Rahim Sweet & Milk Shop Ismatullah Dairy Farm DAIRY Gojar Dairy Farm DAIRY Mithal Dairy Farm DAIRY Madina Bakers & General store Khyber Bakers & Dairies Hussain Bakers & Dairy Shop Near Sui gas colony, sariab mill, Burma hotel, Irrigation colony, Sariab road Chaki shahwani, Sariab road, Village aid, Sariab road,. New Bus adda, Sariab road, Sur pull, Sabzal road, Near Rehmania Madrassa, Arbab Karam Khan road, Killi Tarkha Sabzal road, Near Jhalawan Poultry shop, Podkli chown Sabzal road, Near Farooq flour mills, Arbab Karam Khan Raod,. Hazar Town, Kirani road, TABLE No. Colour, Odour and Temperature noted for the milk samples. S.No Location name Type TEMPERATURE COLOUR ODOUR 1 Wahdat colony, Brewery Road, DAIRY 4 0 C Creamy Cheesy Near Killa Sheikh Hussaini, Brewery road DAIRY 3 0 C Opaque Cheesy 3 Pashtoon Bagh, Kharotabad, Sabzal road, DAIRY 3 0 C Opaque Sweet 4 Golimar chowk, brewery road, 3 0 C Creamy Sweet 5 Near BMC main hostel, brewery road, 5 0 C Creamy Sweet 6 Joint road, 4 0 C Chalky Sweet 7 Near Sui gas colony, sariab mill, DAIRY 3 0 C Chalky Sweet 8 Burma hotel, DAIRY 3 0 C Opaque Cheesy 9 Irrigation colony, Sariab road DAIRY 5 0 C Creamy Sweet 10 Chaki shahwani, Sariab road, 6 0 C Creamy Sweet 11 Village aid, Sariab road,. 5 0 C Chalky Sweet 1 New Bus adda, Sariab road, 4 0 C Opaque Cheesy 13 Sur pull, Sabzal road, DAIRY 5 0 C Opaque Cheesy 14 Near Rehmania Madrassa, Arbab Karam Khan road, DAIRY 5 0 C Opaque Cheesy 3

4 Killi Tarkha Sabzal road, Near Jhalawan Poultry shop, Podkli chown Sabzal road, Near Farooq flour mills, Arbab Karam Khan Raod,. Hazar Town, Kirani road, DAIRY 3 0 C Opaque Sweet 4 0 C Creamy Sweet 3 0 C Opaque Sweet 6 0 C Chalky Sweet TABLE No.3 ph, Water content and Leukocyte count for the milk samples. S.No Location name Type ph WATER CONTENT LEUKOCYTE COUNT Wahdat colony, Brewery Road, Near Killa Sheikh Hussaini, Brewery road Pashtoon Bagh, Kharotabad, Sabzal road, Golimar chowk, brewery road, Near BMC main hostel, brewery road, DAIRY 6.0 ½ DAIRY 6. ½ DAIRY 7. ¾ ½ ½ Joint road, 6.7 ¼ Near Sui gas colony, sariab mill, DAIRY 6.5 ¼ Burma hotel, DAIRY 6.3 ½ Irrigation colony, Sariab road Chaki shahwani, Sariab road, Village aid, Sariab road,. New Bus adda, Sariab road, DAIRY 6. ½ ½ ¼ ¾ Sur pull, Sabzal road, DAIRY 7.5 ¾ Near Rehmania Madrassa, Arbab Karam Khan road, Killi Tarkha Sabzal road, Near Jhalawan Poultry shop, Podkli chown Sabzal road, Near Farooq flour mills, Arbab Karam Khan Raod,. Hazar Town, Kirani road, DAIRY 7.0 ¾ DAIRY 6.1 ½ ½ ½ ¼

5 The quality of milk is concerned very much with the health and life of the consumer. In our country the raw milk that is supplied to the user and the end user is just boiled the milk, but it is still not safe to be use. Colour, Odour and temperature of milk samples The colours of milk samples were observed through comparing them with the colour strip scheme. It was observed from the results of this study that there were three different types of milk colour (Table No. ) i.e. chalky, creamy and opaque. Creamy type of colour ranks top (33.33 %) as compared with the other chalky (.%) and opaque (44.44%) colours. The odour of the milk samples was of two kinds i.e. sweet and cheesy. Out of eighteen samples analyzed 1 samples (66.66 %) had sweet odour while the remaining four (6) samples (33.33 %) had cheesy odour ( Table No. ). The findings of this study are in accordance with the findings of Yadav, et. al. who stated that milk taste and colour are adversely affected due to mastitis, a major disease of the milk secreting organs in which the count of somatic cells increased []. Results regarding the temperature noted for all the milk samples are depicted in Table No.. Which ranged from 3 0 C to 6 0 C at normal room temperature i.e. 0 C. Fig no.1 show the ph concentration of milk of quetta, city. Hydrogen ion Concentration (ph) of milk samples Form the determination of hydrogen ion concentration (ph) of the milk samples, electric ph meter was used. It was observed from the results (Table No. 3) that eight samples (44.44 %) ranged within the range of 6.0 to 6., six samples (33.33 %) ranged within 6.3 to 6.7 vale of ph, while the remaining four samples (. %) lied within the range of of ph meter. The lowest ph (6.0) observed was in two milk samples, while the highest ph was 7.. According to Campbell and Marshall, and Yadav, et.al. ph vale for normal milk ranges from 6.4 to 6.7 and from mastitis milk it ranges from 6.8 to 7.3[, 9]. In current study four samples were of mastitis origin and the remaining sixteen samples show normal milk level. The mean ph is 6.4. Water Contents in milk samples To detect the water content in the milk, Lactometer was used to find the content in the milk samples shows that four(4) samples had ¼, ten (10) samples had 1/, while remaining four (4) samples had ¾ extra water along with the normal contents of the milk. Overall water content shows that % of milk samples comprised of 1/,. % of milk samples had ¾ and. % of milk samples had ¼ extra water, respectively ( Table No. 3). According to Yadav, et. al. (1993) mastitis milk generally has a lower SNF, fat, casein and lactose, which may ultimately be a cause of high water content in milk samples []. Leukocyte Count in milk samples The data regarding the leukocyte count of various milk samples analyzed is shown in Table No. 3. It was observed from the results that the count for milk samples ranged within three main classes of countring i.e. 4 samples within the range of 78,980 to 99,77, 10 samples within the range of 1,0,840 to 1,99,010 while remaining 4 samples ranged with,44,00 to,78,909 cell / ml. The findings revealed that the samples analyzed may be categorized as good, average and bad as per mastitic condition. The mean leukocyte count in 18 was Fig no. Show the leukocytes in milk sample of quetta, city. CONCLUSION Normal milk is that from a health mammary quarter. The presence of flakes, clots or other gross alterations in appearance of quarter milk is evidence of clinical mastitis and is by definition, abnormal milk. Based on the likelihood of infection and altered manufacturing properties, milk from a mammary quarter with a Somatic cell counts equal to or greater than 00,000 cells/ml, with or without clinical signs, is abnormal milk. The predominant factor affecting herd or individual cow s somatic cell count level is intra mammary infections. The causes of infections are many and the method of transmission is virtually never a single factor, but a combination of many factors. Analysis of these factors, combined with herd history, cow environment, and bacteriological culture information provides valuable guidelines for better management decisions that can prevent problems and reduce some misconceptions concerning changes in colour, odour, ph, water content and leukocyte counts. REFERENCES 1. Encyclopædia Britannica, Inc Yadav, J.S, S. Gover, and V.K Batish,. A comprehensive dairy microbiology, Nirala Printers F-19, Navin Shahdra, Delhi. 3.(1993). 3. Dobrzañski Z, Kolacz R, Górecka H, Chojnacka K, Bartkowiak A. The content of microelements and trace elements in raw milk from cows in the Silesian region. Polish J Environ Stud.;14(5): ( 005). 4. Hussain I, Ishaq M, Rehman I, Ahmad I, Shakirullah M. Comparative studies of vitamin C contents in different processed and un-processed milk samples. J Chem Soc Pak.;8(3):36 40(006). 5. Talat Zamir; S. Altaf Hussain Determination of Lead and Cadmium Level in Powdered Milk in (Pakistan) by Atomic Absorption Spectrometry J.Bio science41-413,(001). 6. Brar, R.S H.S Sandhu, and A. Singh, 000 Veterinary Clinical diagnosis by Laboratory methods, Kalyani Publishers B-15, Sector-8, Noida, Utter Pardesh India. Pp The effect of light on the vitamin c of milk by Stanislaw kazimerz kon and Bruce watson. From the 5

6 National Institute for Research in Dairying, University of Reading Received 6 October Harmon, R. J., Physiology of mastitis and factors affecting somatic cell counts. J. Dairy Sci., 77: Campbell J.R and R.T Marshall, The science of providing milk for man. McGraw-Hill Book Company New York Source of support: Nil; Conflict of interest: None declared 6

TANZANIA BUREAU OF STANDARDS

TBS/AFDC 14 (5261) P3 Raw milk Specification DRAFT TANZANIA STANDARD TANZANIA BUREAU OF STANDARDS Raw milk Specification 0 FOREWORD Milk is a nutritious food produced by milking animals for the nourishment