Che m ica l a nd m icr obia l qua lit ie s of m or ning a n d e ve ning m ilk colle ct e d fr om se le ct e d da ir y fa r m s in Chit t a gong Cit y of Ba ngla de sh S Ak t er 1 , M AA Ga lib 2 , H k a bir 1 , SK N a t h 3 , K Afr in 3 , M S Sa r k e r 4 * 1 Depart m ent of Medicine and Surgery, Chit t agong Vet er inary and Anim al Sciences Universit y ,Chit t agong; 3 Depart m ent of Anim al Science and Nut rit ion; 4 Depar t m ent of Microbiology and Vet er inary Public Healt h, Chit t agong Vet erinary and Anim al Sciences Universit y , Khulshi 4225, Chit t agong, 2 Depart m ent of Physiology and Phar m acology, Haj ee Moham m ad Danesh Science and Technology Univer sit y, Dinaj pur, Bangladesh Abst r a ct This st udy was undert aken t o evaluat e t he nut rit ional and m icrobiological qualit ies of raw m ilk in t he Chit t agong Cit y. Eight y sam ples of raw m ilk of cow were collect ed in t he m orning and evening from t he surrounding farm s of Chit t agong Vet erinary and Anim al Sciences Universit y ( CVASU) , Chit t agong, Bangladesh. The result s of com posit ional analysis evolved t hat fat , SNF, prot ein, lact ose, specific gravit y, freezing point and m ineral were ( 3.94± 0.22% ) , ( 7.91± 0.17% ) , ( 3.11± 0.08% ) , ( 4.32± 0.10% ) , ( 1.026± 0.00) , ( - 0.46± 0.007) , ( 0.68± 0.01% ) , respect ively. The fat % of evening m ilk was significant ly higher ( p‹ 0.001) t han m orning m ilk. Rest of t he param et ers were not significant . Microbiological enum erat ion revealed for t he count s of t ot al viable bact eria ( cfu/ m l) and coliform ( cfu/ m l) were 730500± 639 and 280± 1.36 in t he m orning m ilk and 628625± 566 and 326± 1.84 in t he evening m ilk, respect ively. The result s of t he current st udy indicat ed t hat t he nut rit ional qualit y of evening raw m ilk was higher t han in t he m orning raw m ilk. Ke y w or ds: Morning m ilk, evening m ilk, qualit y, Chit t agong Cit y Bangladesh Anim al Husbandr y Associat ion. All r ight s reser ved. I n t r odu ct ion Ba n g. J. An im . Sci. 2 0 1 5 . 4 4 ( 3 ) : 1 7 1 - 1 7 5 Chem ical and m icrobiological analysis is an Milk has been considered as ideal food for hum an im port ant t ool t o m onit or t he qualit y of dairy since long before t he recorded hist ory. Milk is product s. renowned as an ‘alm ost com plet e’ as well as spoilage of dairy product s and t o ensure t he nat ural nut rit ious food for all m am m als including qualit y of m ilk requires essent ial and higher hum an being ( Debnat h et al. 2014) . According t o im port ance Byron et al. ( 1974) , t he const it uent s of m ilk are m et hods wat er ( 87.20% ) , Prot ein ( 3.50% ) , Fat ( 3.70% ) , m onit oring, product ion and processing of m ilk. m ilk sugar or lact ose ( 4.90% ) , ash ( 0.70% ) and Fresh m ilk considered as a com plet e diet because Dry m at t er ( 12.80% ) . These com ponent s m ay it cont ains t he essent ial nut rient s as lact ose, fat , fluct uat e by t ype of breed, t ype of feed t hey prot ein, m ineral and vit am ins in balanced rat io as consum ed, st age of lact at ion, age of cow, season com pared et c and bet ween t he individuals of sam e breed Yadollahi, 2008) . Cousin ( 1982) st at ed t hat t here ( Ghosh et al. 1965) . Milk is t he m ost desirable were a lot of sources for bact erial cont am inat ion food of high biological value and it present s of m ilk like udder, skin surface, lit t er, floor, flies, alm ost all ingredient s of food in exact proport ion insect s and rodent s, wat er supply, m ilker’s hand, and in any digest ible form . I t is im perat ive t o ut ensils, at m osphere et c. Oliver et al. ( 2005) st udy m ilk fat - prot ein rat io ( F/ P) in order t o revealed t hat m ilk and m ilk product s from raw assess m ilk can port a range of m icroorganism s and can nut rit ion, nut rient s conversion and Milk on have to adult erat ion regulat ory to t he t ake ot her can causes aspect s. for foods t he t he Advanced analysis, ( Shoj aei and m et abolism . The opt im um F/ P rat io is 1.2–1.4 be vit al sources of food borne diseases. ( Vladim ír et al. 2005) . Richardt ( 2004) , confirm s Public healt h aut horit y is em ployed t he st andards t hat dict at es of m ilk and dairy product s based on Bangladesh subclinical ket osis whereas t he rat io lower t han St andards ( BDS) . So it is im port ant t o evaluat e 1.1 t hat suspect ed rum en acidosis. nut rit ional and m icrobiological qualit y of raw cow * t he F/ P rat io higher t han 1.5 Corresponding aut hor: sa m u n _ cva su @ya h oo.com 171 Ch e m ica l a n d m icr obia l qu a lit ie s of m ilk m ilk. Therefore, t he present st udy was carried colonies from an individual dilut ion was m ult iplied out in order t o evaluat e t he nut rit ional and by t he dilut ion fact or t o obt ain t he TVC and t ot al m icrobial qualit ies of m orning and evening m ilk of count was expressed as colony form ing unit s per cow collect ed from t he surrounding farm s of m illilit er ( cfu / m l) . CVASU. Colifor m coun t M a t e r ia ls a n d M e t hods The m edium used for coliform was Violet Red Bile agar ( VRB) . I noculat ed plat es were incubat ed at Pla ce a n d D u r a t ion 37° C for 24 hours. Then, t ypical pinkish and red The st udy was carried out in Dairy science Laborat ory belongs t o t he Depart m ent of Dairy colonies were count ed and t ot al coliform was calculat ed. and poult ry Science at Chit t agong Vet erinary and Anim al Sciences Universit y ( CVASU) , during a period from 5t h Sept em ber t o 10t h Oct ober D a t a r e cor din g a n d a n a lysis SPSS ( Version- 16) st at ist ical soft ware was used 2013. for Collect ion of m ilk sa m ple correlat ion t he calculat ion of m ean, coefficient ® , st andard error, t - t est and significant value ( p) . A t ot al num ber of 80 sam ples of raw m ilk were Re su lt s collect ed in t he m orning ( 40 sam ples) and in t he evening ( 40 sam ples) . All t he sam ples were collect ed from surrounding cam pus ( Jalalabad, farm s Foy’s lake of CVASU area) . The nut rit ional and m icrobiological qualit ies of each sam ple were analyzed. Raw m ilk sam ples were collect ed from t he bulk sources of fluid raw m ilk t hrough proper m ixing wit h t he help of plunger and dipper at m orning and evening from each farm . Soon aft er collect ion t he sam ples were kept The average nut rit ional com posit ion of raw m ilk is shown in Table 1. The fat , SNF, prot ein, lact ose, specific gravit y, of acid producing organism s point and m ineral cont ent of raw m ilk from different farm s were 3.94± 0.22% , 4.32± 0.10% , 7.91± 0.17% , 1.026± 0.00, 3.11± 0.08% , - 0.46± 0.007 and 0.68± 0.01% , respect ively. Ta ble 1. Average nut rit ional com posit ion and t ransport ed t o t he laborat ory. Param et ers Mean± SE M e t h ods for t e st in g r a w m ilk Fat 3.94± 0.22 SNF 7.91± 0.17 Prot ein 3.11± 0.08 conduct ed. Before analysis each sam ple was pre Lact ose 4.32± 0. 10 warm ed Sp. Gravit y 1.026± 0.0008 Freezing point - 0.46± 0.007 Mineral 0.68± 0.01 The collect ed m ilk sam ples were kept in t he refrigerat or at for chem ical 40° C few unt il m inut es propert ies t he at t est s 600° C. ( specific were Physio- gravit y, fat percent age, SNF percent age, prot ein percent age, lact ose percent age, Freezing point and Mineral percent age) were det erm ined by of different farm raw m ilk ( Mean± SE) int o t he ice box for ceasing t he growt h and act ivit y freezing Lact ost er N= 80 ( 40 at m orning + 40 at evening) m achine ( Germ any) . The Tot a l Via ble Cou n t ( TVC) m orning m ilk com posit ion of fat , SNF, prot ein, lact ose, specific gravit y, freezing point TVC were done as per recom m endat ion of APHA ( 1967) . In brief, Bact o agar was used for enum erat ion of SPC. Each raw m ilk sam ple was prepared by t en fold dilut ion using pept one buffered wat er. To det erm ine t he SPC, using st erile pipet t e t o t ransfer and t o spread of 0.1 m l of each dilut ion on agar surface. I noculat ed plat es were incubat ed at 37° C for 48 hours t o facilit at e viable bact erial growt h. Aft er incubat ion, t he 172 colonies were count ed. The num ber of and m ineral were 3.12± 0.05% , 7.63± 0.27% , 2.97± 0.11% , 4.13± 0.15% , 1.026± 0.001, - 0.45± 0.009 0.69± 0.01% , respect ively ( Table 2) . On t he ot her hand, t he evening m ilk com posit ion of fat , SNF, prot ein, lact ose, specific gravit y, freezing point and m ineral were 4.75± 0.11% , 8.19± 0.17% , 3.25± 0.08% , 4.50± 0.10% , 1.027± 0.0009, - 0.47± 0.007 and 0.67± 0.02% , respect ively ( Table 2) . I n t he m orning m ilk, TVC and Coliform count were 730500± 639 and Ak t e r e l a l. ( 2 0 1 5 ) Ba ng. J. Am in . Sci. 4 4 ( 3 ) : 1 7 1 - 1 7 5 280± 1.36, respect ively. But in t he evening m ilk it prot ein rat ion of m orning and evening m ilk were was 628625± 566 and 326± 1.84 ( Table 3) . Fat 1.05 and 1.46, respect ively ( Table 4) . Ta ble 2 . Nut rit ional com posit ion of m orning and evening raw m ilk of different farm Morning Fat SNF Prot ein Lact ose Sp. Gravit y F. point Mineral 3.12± 0.05 7.63± 0.27 2.97± 0.11 4.13± 0.15 1.026± 0.001 - 0.45± 0.009 0.69± 0.01 4.75± 0.11 8.19± 0.17 3.25± 0.08 4.50± 0.10 1.027± 0.001 - 0.47± 0.007 0.67± 0.02 0.001 0.159 0.069 0.87 0.507 0.20 m ilking Evening m ilking Significant 0.112 value N= 80 ( 40 at m orning + 40 at evening) Ta ble 3 . Microbial qualit y of different farm raw m ilk TVC ( CFU/ m l) Coliform ( CFU/ m l) Morning m ilking 730500± 639 280± 1.36 Evening m ilking 628625± 566 326± 1.84 Significant value 0.150 0.082 N= 80 ( 40 at m orning + 40 at evening) Ta ble 4 . Fat - prot ein rat io of m ilk Morning m ilk Fat Prot ein Fat - prot ein rat io 3.12 2.97 1.05 Evening m ilk 4.75 3.25 1.46 Morning and evening m ilk 3.94 3.11 1.26 D iscu ssion 3.80% prot ein and 4.80% lact ose. This variat ion m ay be due t o t he variat ion of breed. The Ph ysicoch em ica l pr oper t ie s The average fat percent age of farm m ilk was found t o be 3.94± 0.22% , which was support ed by Debnat h et al. ( 2009) who dem onst rat ed t hat t he but t er fat of m ilk from different sources from Chit t agong m et ropolit an area varied from 3.52 t o 4.01. Judkins and Keener ( 1960) report ed t hat t he average fat % of m ilk sam ple was 2.5 t o 8.0% . The st udy indicat ed t hat t he average SNF cont ent of farm m ilk was 7.91± 0.17% , which was slight ly lower t han t he findings of Debnat h et al. ( 2009) who report ed t hat SNF% from farm produced m ilk, vendor supplied farm m ilk, vendor supplied rural m ilk and brand m arket m ilk were 8.33, 7.98, 7.85, 8.2% , respect ively in Chit t agong m et ropolit an area. The slight ly lower SNF value m ight be due t o sm aller sam ple size. I n t he present st udy, t he average prot ein and lact ose cont ent of farm m ilk were 3.11± 0.08% and 4.32± 0.10% , respect ively. These values were lower t han t he values of Eckles et al. ( 1951) , dem onst rat ed t hat m ilk should cont ain average specific gravit y of farm m ilk was 1.026± 0.00, which was support ed by Eckles et al. ( 1951) , I slam et al. ( 1993) and Debnat h et al. ( 2009) . The average freezing point and m ineral cont ent of farm m ilk were - 0.46± 0.007 and 0.68± 0.01% , respect ively. According t o Eckles et al. ( 1951) , m ilk should cont ain 0.65 % m inerals and t he current st udy agreed wit h his finding. The average fat percent age of evening m ilk was significant ly ( P < 0.05) higher t han t he m orning m ilk. I t was support ed by I qbal et al. ( 2005) . The average SNF, prot ein, lact ose, specific gravit y, freezing point and m ineral percent age of evening m ilk was not significant ly ( p > 0.05) higher t han t he m orning m ilk. This st udy was support ed by I qbal et al. ( 2005) . M icr obia l qua lit y The average TVC count of m orning m ilk ( 730500± 639) was not significant ly higher t han t he evening m ilk ( 628625± 566) ( Table 3) . The result s of t his st udy correlat ed wit h t he findings 173 Ch e m ica l a n d m icr obia l qu a lit ie s of m ilk of I knom ov et al. ( 1956) who report ed t hat t he hazardous product which is likely t o be posing a t ot al bact erial count s ranged from 1, 25,000 t o serious healt h risk t o t he consum ers. Fat prot ein 9,000,000 per m l of m ilk depending on m ilking rat io was in accept able range. Evening m ilk was t echniques of found superior t han m orning m ilk due t o high m ilk was 100- chem ical com posit ion. The governm ent t herefore infect ion occurred and cleanliness. bact eria in asept ically 92,000 per m l, The drawn but num ber should conduct frequent inspect ion of t he subsequent ly from t he skin of anim als, m ilker’s m arket ed m ilks t o check whet her t hey m eet t he hand, cow shed and m ilking ut ensils. Lee et al. m inim um legal st andards and should m onit or t he ( 1983) conduct ed an experim ent in Seoul of overall Korea and found t hat t he bact erial count in raw product ion m ilk ranged from 4 × 106 t o 2.7 × 107 per m l. st andards for t he raw m ilks need t o be devised This result indicat ed t hat t he average coliform and appropriat e t raining should be given t o t he count raw m ilk producers in hygienic handling of m ilk. of m orning m ilk was lower t han t he hygienic condit ion and handling surrounding of m ilk. t he Realist ic evening m ilk but not st at ist ically significant . The Ack now le dge m e n t s variat ion in coliform count of t he m ilk m ay be due t o t he hygienic m aint enance during m ilking. The result s of t his invest igat ion were in agreem ent wit h t he finding of Mut ukum ira et al. ( 1996) , who found t he coliform bact eria was 326 t o 240 per m l. Sait anu et al. ( 1996) experim ent ed and Aut hors are grat eful t o t he academ ician and st uffs of Depart m ent of Dairy and Poult ry Science ( CVASU) and Owner of t he respect ed farm s. revealed t hat t he t ot al coliform count was of Re fe r e n ce s < 1000 CFU/ m l. Am erican Fa t - pr ot ein r a t io Public ( 1967) . Healt h St andard Associat ion m et hods ( APHA) for t he exam inat ion of dairy product s. Am erican The fat - prot ein evening m ilk ( F/ P) was rat io of 1.26. m orning Richardt and Public Healt h Associat ion. I nc. 12t h Edn. ( 2004) m easured t he F/ P rat io as an im port ant point er of New York. P. 90 - 110. Byron HW, Arnold HJ, Johan AA ( 1974) . anim al healt h. I t s value above 1.5 in dairy cows Fundam ent als of dairy chem ist ry. 2nd Edn. indicat ed West port , Connect icut . The Avi Publishing a 1.5 t im es m ore probabilit y of incidence of m ast it is, 7.5 t im es t endency t o lam eness and 3.5 t im es m ore incidence to Com pany, I nc. P. 2- 8. Cousin MA ( 1982) . Presence and act ivit y of ket osis. Vladim ir et al. ( 2005) dict at ed t hat t he psychrot rophic m icroorganism s in m ilk and proper F/ P rat io for Holst ein cows is 1.05 - 1.18. dairy product s. Journal Food Prot ein. 45: High values in t he first phase suggest ed a great 172- 207. energy deficiency. Haas et al. ( 2004) included Debnat h GK, Kober AKMH, Chanda GC ( 2009) . t hat t he opt im um F/ P rat io is 1.2–1.4. Low values Qualit y of Fluid Milk Available in Chit t agong are prone t o subclinical rum en acidosis which can Met ropolit an Area, proceedings of t he sixt h put in danger t he reproduct ive perform ance of annual cows Bangladesh. P.178- 182 and increase a possible expansion of scient ific conference of CVASU, disorders of m ineral m et abolism . The F/ P rat io Debnat h GK, Kober AKMH, Chanda T, Chanda higher t han 1.4 indicat ed t he sign of energy GC, Bari MSA ( 2014) . Com parat ive St udy on deficit and if ket one bodies are present , lead t o t he Qualit y of Available Brand and Non subclinical ket osis. The current st udy agreed wit h Brand Fluid Milk Consum ed by t he People of t his range. Chit t agong Cit y of Bangladesh. I nt ernat ional Journal of Nat ural Sciences, 4( 1) : 16- 20. Con clu sion Eckles C, Com bs C, Macy H ( 1951) . Milk and m ilk Result s of t his st udy showed t he chem ical and product s m icrobial qualit ies of m ilk from different local com pany, New York, Toront o, London. P. farm s. The presence of bact erial populat ion in raw m ilk indicat es defect during collect ion and processing of m ilk. The presence of coliform s in m ilk are indicat ive of a pot ent ially 174 Edn. Mcgraw- Hill book 22- 23. Ghosh SN, Krishnan CPA ( 1965) . I ndian Journal t he pat hogenic organism s and t he high count s of 4t h of Dairy Science, 17: 190. Haas D, Hofírek B ( 2004) . The diagnost ic im port ance of m ilk com ponent s for a hum an Ak t e r e l a l. ( 2 0 1 5 ) Ba ng. J. Am in . Sci. 4 4 ( 3 ) : 1 7 1 - 1 7 5 and cow’s healt h. ( I n Czech) CUA Prague, SP, Jayarao BM, Alm eida RA ( 2005) . Foodborne pat hogens in m ilk and t he dairy – 29. farm environm ent : food safet y and public I konom ov L, Lot ov I , Todorov D, Tankov G, Dzhurov TS ( 1956) . Bact eriological st udies of Hygiene of m ilk product ion of Bulgarian cat t le Breeding Farm s. SA, Mido Discovery Y healt h im plicat ions. Foodborne Pat hogen and Diseases 2 ( 2) : 115- 29. Richardt W ( 2004) . Milk com posit ion as an Science indicat or of nut rit ion and healt h. ( I n Czech) Chem ist ry. Sait anu I A, Chuanchuen KR, Nuanuarsuwan S, Dairy The Breeding. P. 26 –27. Abst ract 19: 936. I qbal Oliver Proceedings of cont ribut ions: Milk day. P. 26 ( 2005) . Publishing Food House, New Delhi, Koowat ananukul C and Rugkhaw V ( 1996) . Microbiological qualit y of raw cow m ilk. Thai I ndia. I slam MN, Hussain SMI , Mannan AKMA ( 1993) . St udies on t he physical param et ers and chem ical qualit ies of m arket m ilk in Journal of Vet erinary Medicine 26( 3) : 193214. Shoj aei ZA, Yadollahi A ( 2008) . Physicochem ical Mym ensingh t own ( Bangladesh) . Banglasesh and j ournal of Anim al Science. 13: 52. Past eurized and UHT Milks in Shops. Asian Judkins HF, Keener HA ( 1960) . Milk product ion and processing. 4t h Edn, New York. P. 227- Microbiological Qualit y of Raw Journal of Scient ific Research. 1: 532- 538. Vladimír ČEJNA, Gustav CHLÁDEK (2005) . The im port ance of m onit oring changes in m ilk 230. Lee JT, Park SY, Korea I K, Kin HU ( 1983) . Qualit y fat t o m ilk prot ein rat io in Holst ein cows of raw m ilk in Korea. Korean Journal of during Dairy Science. 5 ( 1) : 22- 28. Breeding, Mendel Universit y of Agricult ure Mut ukum ira AN, Abraham sen Feresu RK SB, ( 1996) . Narbhus JA, Chem ical and lact at ion, Depart m ent of Anim al and Forestry in Brno, Zemědělská 1, 613 00 Brno, Czech Republic. Microbiological Qualit y of raw m ilk produced by sm all holder farm ers in Zim babwe. Journal of Food Prot ect ion. 59 ( 9) : 984- 987. 175