Změny aminokyselinového spektra kravského mleziva v průběhu prvních 72 hodin po porodu

ACTA UNI VERS ITATIS AGRICULTURAE ET SILVICULTURAE MENDELIANAE BRUNENSIS SBORNI K MENDELOVY ZEMEDELSKE A LESNICKE UNIVERZITY V BRNE Rocn k LV 10 Cislo 1, 2007 CHAN GE S INAMINOACIDS COMPOSITION OF COWS COL OST RUM(DURING FIRST 72 HOURS AFTER PARTURITION) S. K acma ,F.Bu ika,I. Hoza, L. Cecho a, P. Valasek Recei ed: Oc obe 9,2006 Abs ac KRACMAR, S., BuNKA , F., HOZA , 1. , CECHOV A, L., VALAsEK, P.: Changes in amino acids com- posi ion o cows colos um (du ing i s 72 hou s a e pa u i ion). Ac a uni . ag ic. e sil ic. Mendel. B un., 2007, LV, No.1 , pp. 81-94 Changes in amino acid spec um o colos um wi hin he pe iod o 2 o 72 hou s pos pa um we e s u- died in dai y cows o he Czech Red Pied ca le wi h a gene ic admix u e o Black Pied Low land b eed (n =10). Fi e o hem we e on he 1s lac a ion and 5 on he 2nd and/o highe lac a ions.Wi hin he s udy pe iod, he con en s o non-essen ial amino acids (NEAA) and essen ial amino acids (EAA) dec e- ased by 22.8-63.2% and 27.3-65.2% , esp., in dai y cows on he 1s lac a ion while in hose on he 2nd and ollowing lac a ions he co esponding alues we e 22 .0-63.7% and 24.9- 71.7%, espec i ely. The dependence o he cou se o changes in he AA spec um o cow's colos um on he ime in e al elapsing a e he pa u i ion could be cha ac e ised by a eg ession equa ion y=bo+ b/x + b/x 2• dai y cows, colos um, amino acids, eg ession analysis Milk is a complex biological sys em, in which i s indi idual componen s occu in di e en a ios and in mu ual ela ionships. Basing on esul s o long- e m s udieson milk composi ion o di e en animal spe- cies was possible o de ine a e age con en s o indi- idual milk componen s, hei a iabili y and (in he majo i y o cases) also hei chemical composi ion. C ude p o ein is he mos complica ed componen o bo h colos um and milk. Rega ding hei nu i ional and echnological impo ance, hese a e he mos s u- died milk componen s a all and i can be said ha composi ion o milk p o ein is o a g ea deg ee depen- den on he s age o lac a ion,season, b eed, heal h condi ion o animals and me hod o milking. Indi e en animal species,p oduc ion o colos um and milk is de e mined by a numbe o physiologi- cal p ocesses, which ake place unde comple ely con- c e e condi ions and in dependence on mo he 's geno- ype .P opo ions o indi idual colos um and milk componen s a e ypical o each b eed and each ani- 81 mal species. A good knowledge o milk and colos um composi ion, a iabili y in le els o hei indi idual componen s and ela ionships exis ing among hem enable us o es ima e expec ed changes as induced by in e nal and ex e nal ac o s wi h ag ea e accu- acy (Schablin, 1987; Khalil e al. 1992; Sai o e al., 1993). The le els o selec ed biochemical pa ame e s (i.e. p o eins, a , lac ose, i amins A and E, and immuno- globulins) in cow 's colos um a he momen o he i s milking a e pa u i ion we e s udied by Jagos e a . (1985) and Bouda e al. (1988); in goa s and sheep asimila s udy was pe o med by K ac na e al. (2003, 2005). The aim o his s udy was o ob ain deepe and mo e exac da a abou he composi ion o cow's colos um wi hin he pe iod o he i s 72 hou s pos pa um in dai y cows on he i s as well as on he second and ollowing lac a ions. 82 S. K acm a ; F. Bunka,IHoza, L. .Cecho a, P. Valasek MATERIAL AND METHODS Changes in he nu i i e alue o colos um wi hin he in e al o 2- 72 hou s a e he pa u i ion we e ollowed in dai y cows o he Czech Red Pied b eed wi h an admix u e o blood o Black Pied Lowland b eed (n =10). Fi e o hem we e on he i s lac a ion and i e on he second and ollowing lac a ions. The a e age li e body weigh s o p imipa ae emales and o dams on he 2nd and ollowing lac a ions we e 500 kg and 550 kg, espec i ely.The s udy was ca ied ou du ing he win e season om No embe o Ma ch. The composi ion o eeding a ions, hei nu i i e alue as dependen on he ank o lac a ion, me hod o milk sampling, sample p epa a ion and sample p ese - a ion we e desc ibed in ou ea lie pape (K aeme & Zeman, 2003). Expe imen al animals did no show any heal h p o- blems du ing he whole s udy pe iod. Colos um samples o amino acid de e mina ion we e adjus ed using acidic and oxida i e acidic hyd- olysis HCl (c =6mol/dm"), The ch oma og aphic analysis o sample hyd olysa es was pe o med in he AAA 400 analyse (manu ac u e INGOS P ague, CR) and using Na-ci a e bu e s and ninhyd in de- ec ion (K acma e al., 1998) as published in he O - icial Jou nal L 206 (Anonymus, 1978) All esul s we e e alua ed using he ANOVA a i- a ion s a is ics p og amme. Co ela ion ma ices and eg ession unc ions we e calcula ed acco ding o Sne- deco and Coch an (1967) when using he s a is ical package Unis a , . 5.I and O ice Excel®Mic oso . RESULTS AND DISCUSSION The ob ained esul s we e analysed sepa a ely o he g oup o p imipa ous dai y cows and o hose on he 2nd and ollowing lac a ions. A e age con en s o essen ial and non-essen ial amino acids, sum o amino acids (~A A ) , sum o essen- ial amino acids (~EA A) , sum o non-essen ial amino acids ( ~ N E A A) and sum o Me +Cys ( ~M e + Cy s ) in colos um o p imipa ous emales wi hin he i s 2 o 72 hou s pos pa uma e p esen ed in Tabs I and II. When compa ing alues eco ded 12;24; 36;48 and 72 hou s pos pa um wi h hose ound ou 2 hou s a - e pa u i ion (= 100%), he ollowing conclusions we e d awn: Du ing he pe iod o in es iga ion, le els o NEAA and EAA dec eased wi hin anges o 22 .8-63 .2% and 27.3 -65.2% , espec i ely. When compa ing he dec ease in con en s o EAA wi hin he pe iod o 12 h wi h he ini ial 100% le el (i.e. 2 hou s pos pa um), he ollowing esul s we e ob ained: Val >10%; His >25%; Phe, A g, and Th >30%; Me ; Lys, Leu, and He >35%, and Cys 50%. The co esponding dec eases wi hin he in e al o 24 h we e as ollows: Val >35%; His >40%; Phe, A g, He, Th , and Leu >50%; Me and Lys >55%, and Cys >65%. Wi hin he in e al o 36 h, he ollowing dec eases we e obse ed: Val >45%; His >50%; Phe, A g and Leu >55%; Me , He, Lys and Th >60%, and Cys >80%. A e 48 h, he dec ease in EAA le els was as ollows: His and Val >45%; A g, Phe, He, Th , Leu and Lys >55%; Me >65%, and Cys >80%.A e 72 h, he ollowing esul s we e ob ained: Val >60%; His, Leu and Lys >65%; Phe, A g, Th and He>70%; Me >75%, and Cys >85%. When compa ing he dec ease in con en s o NEA A wi hin he pe iod o 12 h wi h he ini ial 100% le el (i.e. 2 hou s pos pa um), he ollowing esul s dec e- ases we e obse ed: Glu >20%, Se , Gly and Ty >25%; Asp and Ala >30%, and P o >35%. When compa ing con en s o NEAA a e 24 wi h he ini- ial alues, he dec eases we e as ollows : Gly >25%; Ala >30%; Se and Ty >35%; Asp >40%, and Glu and P o >45%. A e 36 h, con en s o NEAA dec e- ased as ollows: Ala >45%; Asp, Se and Ty >50%; Glu and P o >55%, and Gly >60%. A e 48 h, he dec ease in EAA le els was as ollows : Gly and Ala >40%; Ty >45%; Asp and Se >50%, and Glu and P o >55%. Con en s o Asp, Glu, Ala and P o dec ea- sed by >65%. Finally, he dec ease in con en s o Se , Gly, and Ty a e 72 h was >70%. A e age con en s o essen ial and non-essen ial amino acids, sum o amino acids (~AA), sum o essen- ial amino acids (~EAA) , sum o non-essen ial amino acids (~NE AA) and sum o Me +Cys (~Me +C y s) in colos um o dai y cows on he 2nd and subsequen lac a ions a e p esen ed in Tabs III and IV. When compa ing alues eco ded 12; 24; 36; 48 and 72 hou spos pa um wi h hose ound ou 2 hou s a - e pa u i ion (= 100%), he ollowing conclusions we e d awn: Du ing he pe iod o in es iga ion, le els o NEAA and EAA dec eased wi hin anges o 22 .0-63.7 % and 24.9 -71.7% , espec i ely. When compa ing he dec ease in con en s o EAA wi hin he pe iod o 12 h wi h he ini ial 100% le el (i.e. 2 hou s pos pa um), he ollowing esul s we e ob ained: Val >5%; His >25%; Phe, A g, and Th >30%; Me , A g, Ile and Leu >30%; Leu >35% and Cys >45%. The co esponding dec eases wi hin he in e al o 24 h we e as ollows : Val >45%; Ty >50%; Phe, Me , His and Leu >55 %; A g, Heand Lys >60%, and Cys >70%. A e 36 h, con en s o EAA dec eased as ollows: Val >40%; His >50%; Me , Phe, A g, Ile, Th and Leu >55%, Lys >60% and Cys >70%. Ala >45%; Asp, Se and Ty >50%; Glu and P o >55%, and Gly >60%. A e 48 h, he dec ease in EAA le els was as ollows: Val >50%; His >55%; Phe, A g, Leu and Lys >60%, Me , Heand Th >65%, Changes in amino acids composi ion o cows colos um (du ing i s 72 hou s a e pa u i ion) 83 and Cys >70%. Gly and Ala >40%; Ty >45%; Asp and Se >50%, and Glu and P o >55%.A e 72 h, he ollowing dec eases in con en s o NEA A we e eco - ded: Val >65%, Lys >70%, Phe, A g, Th , Me , Leu and lIe >76%, and His and Cys >76%. As compa ed wi h he e e ence alue (i.e. ha e- co ded 2 h pos pa um), he con en s o NEAA ob- se ed in cow's colos um a e 12 h dec eased in he ollowing manne : Glu >20%, Se , Gly, Ala and Ty >25%, and Asp and P o >30%. When compa ing con- en s o NEAA a e 24 h, he dec eases we e as ol- lows: Gly, Ty , Asp , Se , Glu and Ala >48 %, and P o >51%. Wi hin he in e al o 36 h, le els o NEAA dec eased as ollows: Ala, Gly, Asp and Ty >46%; Se >47%, and Glu and P o >51%. A e 48 h, he de- c ease in NEAA le els was as ollows: Ala and Ty >50%; Gly and Ala >40%; Ty >45%; Asp and Se >50%, and Glu and P o >55%. A e 72 h, he ollow- ing dec eases in con en s o NEAA we e ound ou : Asp >65%; Glu, Ala, Ty , Se and P o >70% and Gly >72%. A e age con en s o essen ial and non-essen ial amino acids, sum o amino acids (LAA), sum o essen- ial amino acids (LEAA) , sum o non-essen ial amino acids (LNEAA) and sum o Me +Cys (LMe +Cys) in colos um o dai y cows wi hin 2 o 72 h pos pa um a e p esen ed in Tabs V and VI and in Figs 1-4. When compa ing he dec ease in con en s o indi- idual amino acids wi hin he ime in e al o 12 h pos pa um wi h he ini ial alue ound 2 h a e he pa u i ion (100 %) i was concluded ha in p imipa- ous dai y cows his dec ease in con en s o EAA was always s eepe han in dai y cows on he 2nd and sub- sequen lac a ions. Wi h excep ion o Me and Lys, a simila endency was ound ou also 72 h pos pa um. Essen ial amino acids His, Phe, A g, lIe, Th , and Val showeda simila end 36 h pos pa um.As a as Leu was conce ned, an iden ical endency was ound ou inall dai y cows 12; 24; and 36 h pos pa um, while in Lyswi hin he in e al o 24 o 72 h and in Me 12, 24, and 72 h a e he pa u i ion. In case o Cys no ag eemen was ound ou in any ime in e al. As compa ed wi h olde dai y cows, in p imipa ae he dec ease in con en s o AA was always s eepe : as compa ed wi h he con en s eco ded 2 h a e pa u- i ion (100%), lowe Cys le els we e eco ded 12; 36 and 72 h, in case o Me 36 and 48 h, and in case o His, Phe, A g , lIe, Th , Val, and Lys 12 h pos pa um. Alowe dec ease in he le el o Cys was obse ed 24 and 48 h pos pa um, in le els o His, Phe, A g, lIe, Th and Val 24 o 72 h pos pa um and in ha o Leu 48 and 72 h pos pa um. As a as NEAA we e conce ned, asimila dec ease in he le el o Gly was ound ou a e 12 h, in le els o Asp and Ty a e 12 and 72 h, in le els o Se and Ala a e 12; 36 a 72 h, in he le el o Glu a e 12; 48 and 72 h and in ha o P o 12 and 36 o 72 hpos pa - um. In p imipa ous dai y cows a as e dec ease was eco ded in le els o Asp , Glu, Gly and Ty 36 h pos pa um and a slowe dec ease in le els o Glu and P o 24 h pos pa um, in hose o Asp, Ala, Ty and Se 24 and 48 h pos pa um and in ha o Gly 24; 48 and 72 h pos pa um. K aeme e a . (1997) concluded ha 12 h pos pa - um he a e age con en o amino acids dec eases by app oxima ely 25% as compa ed wi h alues ound ou 2 h a e he pa u i ion; a e 24; 36 and 48 hou s, hei con en s dec eased by ca 50% , 52% , and 54- 60%, espec i ely.Simila esul s we e ob ained also in his expe imen . The dependence o changes in con en s o amino acids in colos um o p imipa ous and/o olde dai y cows wi hin di e en ime in e als pos pa um can be exp essed by he eg ession equa ion y=bo+ b /x + b /x 2•Agg ega e alues a e p esen ed in Tabs V II- IX. Con en s o amino acids in milk p o ein show ahigh deg ee o a iabili y. Acco ding o Polansky (1988), con en s o amino acids in colos um and milk can be in luenced by quali y o eeds (e.g. by silage o in e- io quali y) as well as by some o he ac o s. Limi alues obse ed in his s udy co esponded wi h da a published by Sebela e a . (1976),Bo os (1988),G ie- ge e a . (1990) and Zeman e a . (1995) o he con- di ions o he Czech Republic and o Slo akia. I: Dynamics o changes in con en o essen ial amino acids in colos um ojp im ipa ous dai y cows wi hin he i s 2- 72h a e pa u i ion (mg/IOOml) 00 .I>- THR YAL ILE LEU PHE hou mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) 2761.1 ± 31.37 9.2 637.2 ± 35.82 16.0 668.0 ± 22.98 7.7 1146.0± 27.75 5.4 6 5.2 ± 24.60 8.9 12 564.7 ± 36.08 4.6 663.4 ± 23.94 8.1 448.3 ± 22.52 6.2 776.0 ± 24.03 6.9 440.4 ± 26.98 13.7 24 395.4 ± 23.9 13.5 4 77.5±48.13 22.5 342.5 ± 35.83 23.4 569 .6± 17.71 7.0 3 5.3 ± 29.86 3.5 36 363.8 ± 28.87 7.7 432.9 ± 29.83 15.4 309.0 ± 32.84 23.8 536.3 ± 35.06 14.6 277.6 ± 27.58 3.8 48 366.7 ± 28.47 14.2 4 4.3 ± 38.05 20.5 307.5 ± 31.14 22.6 542.7 ± 38.60 15.9 300.0 ± 22.68 16.9 !=':l 72 238.1 ± 24.9 23.4 3 2 .0 ± 33.20 23.8 204.1 ± 40.6 24.5 436.0 ± 25.05 12.8 198.3 ± 28.10 13.7 ~ ~, ~ ' ~ , "" HIS LYS ARG CYS MET ~ hou b:l mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) ~ ;:s. 2466.3 ± 37.27 7.9 1004.4 ± 35.95 8.0 639.8 ± 69.48 0.5 310. ± 16.40 11.8 237.4 ± 13.39 13.8 "'"" ,~ 12 353.7 ± 38.03 24.0 691.8 ± 25.36 8.2 45 .3 ± 44.54 12.2 161.0 ± 14.13 5.7 164.7 ± 14. 9 ~ 15.7 ~ 24 277.6 ± 21.67 7.5 492.4 ± 21.62 9.8 329.5 ± 33. 0 22.5 108. 7± 1.43 12.9 16.7 ± 1 I.I 8 12.3 '" , ~ 36 237.7 ± 22 . 6 20.8 445.2 ± 24.83 2.5 303.4 ± 16.86 12.4 73.9 ± 1.04 13.1 102.4 ± 10.74 16.0 !:""" 48 254.8 ± 20 . 8 7.6 444.8 ± 39.74 20.0 303.7 ± 33.99 25.0 82. ± 1.93 3.3 86.2 ± 8.78 21.8 (J . Cll 72 161.1 ± 13.38 14.7 369.6 ± 7.42 10.5 222.3 ± 24.67 24.8 50.9 ± 6.05 12.6 62.9 ± 24.56 18.7 s c ~ ,~, ~ LCYS+MET LEAK LAK ~ ..... hou ~, ">. mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) Cll "'"" 2 547.5 ± 57.54 10.5 6485.4 ± 11 0.2 8.9 12448.2± 80.50 9.0 12 325.7 ± 14.81 13.3 4715.3 ± 32.08 16.8 9319 .6± 115.80 13.8 24 225.4 ± 12.26 12.2 3425.1 ± 60.97 14.0 7132.4 ± 129.73 19.0 36 176.4 ± 7.65 9.7 3082.3 ± 92.90 6.7 6059.3 ± 121.16 18.0 48 168.2 ± 2.96 17.2 3102.7 ± 144.44 10.4 6155.7 ± 123.51 18.6 72 113.9 ± 12.78 15.5 2255.4 ± 68.10 16.3 4448.9 ± 63.38 14.2 II: Dynamics o changes in con en o non -essen ial amino acids in in colos um o p imipa ous dai y cows wi hin he i s 2- 72 h a e pa u i ion (mg /JOO ml) ASP SER GLU PRO hou CV(%) CV(%) mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D.mean±S.D. 2802.9 ± 35.62 9.9 677.4 ± 15.11 5.0 2252.1 ± 311.62 30.9 991.0 ± 22.31 5.0 12 598.1 ± 50.77 18.9 534.5 ± 25.51 10.7 1860.9 ± 177.88 21.4 695.6 ± 45.82 14.7 24 427.8 ± 30.42 15.9 366.0 ± 20.52 12.5 1262.6 ± 171.40 30.4 488.5 ± 29.63 13.5 36 433.9 ± 44.71 23.1 363.4 ± 41.18 25.3 1183.8 ± 255.27 48.2 494.0 ± 42.78 19.3 48 395.7 ± 27.68 15.6 312.5 ± 29.17 20.8 1078.7 ± 199.65 41.4 470.1 ± 50.59 24.1 72 296.6 ± 30.44 22.9 231.5 ± 31.19 30.0 850.1 ± 32.10 8.4 387.4 ± 30.02 17.5 GLY ALA TYR ~ NEAK hou mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) 2 284.8 ± 35.74 28.1 374.4 ± 22.43 13.4 552.6 ± 32.61 13.2 5935.2 ± 104.71 10.3 12 227.3 ± 22.63 22.3 281.7 ± 25.11 19.9 430.0 ± 51.68 26.8 4628.0 ± 83.25 8.5 24 154.4 ± 26.01 37.7 203.4 ± 36.33 40.5 288.3 ± 30.62 23.7 3 191.0± 27.96 19.6 36 154.1 ± 18.63 27.1 219.9 ± 23.81 24.2 308.5 ± 26.38 19.1 3157.6 ± 30.65 12.7 48 138.0 ± 33.91 41.8 191.2 ± 8.03 9.7 279.3 ± 21.19 16.9 2865.5 ± 31.56 14.6 72 81.2 ± 13.25 10.3 130.6 ± 16.63 8.5 176.6 ± 11.52 6.0 2153.7 ± 10.98 11.4 S' 3 S' o ~ 0: en (') o 3 '0 o ~ . .... o' e o ...., (') ~ en (') o 0' en ::: s:: 3 III: Dynamics o changes in con en o essen ial amino acids in colos um o dai y cows on he 2nd and subsequen lac a ions wi hin he i s 2 -72 h a e pa u i ion 00 (mg/lOO ml) 0'1 THR YAL ILE LEU PHE hou mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) 2759.5 ± 34.47 10.1 725.1 ± 33.13 10.2 659.7 ± 39.60 13.4 1150.5 ± 26.52 5.2 608.2 ± 21.15 7.8 12 593.9 ± 29.31 11.2 689.6 ± 13.52 4.4 471.2 ± 22.65 10.7 795.0 ± 31.77 8.9 456.8 ± 20.16 9.9 24 366.1 ± 47.31 28.9 432.3 ± 23.84 12.3 288.6 ± 48.71 13.7 522.8 ± 26.36 11.3 273.9 ± 28.72 23.5 36 354.5 ± 32.80 20.7 435.4 ± 20.89 10.7 298.4 ± 38.08 26.5 525.8 ± 21.10 8.9 281.3 ± 37.76 18.4 48 303.0 ± 31.73 18.8 372.5 ± 31.40 18.8 260.8 ± 32.79 18.3 494.7 ± 31.14 14.1 258.4 ± 28.35 15.5 Y:l ~ 72 189.5 ± 30.25 13.6 264.9 ± 30.66 25.9 160.0 ± 31.25 43.7 338.7 ± 59.I929.5 149.0 ± 10.14 15.2 l::l. ~ ' l::l ..... ~ HIS LYS ARG CYS MET l hou :::: ;:s, mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) ;:.;- .l::l 2459.4 ± 16.47 8.0 993.4 ± 44.43 10.0 631.7 ± 59.24 21.0 290.2 ± 34.86 26.9 229.3 ± 51.11 26.9 :--. 12 359.6 ± 36.59 22.8 724.6 ± 29.23 9.0 466.0 ± 23.18 11.1 166.9 ± 41.84 36.1 165.1 ± 64.02 18.7 ~ N 24 218.5 ± 88.11 9.0 443.7 ± 60.39 30.4 284.0 ± 40.05 21.8 77.8 ± 17.45 3.8 114.1 ± 11.21 Fl 5.9 ~ 36 232.3 ± 45.47 21.5 438.3 ± 30.94 15.8 297.3 ± 26.09 19.6 75.8 ± 6.36 1.8 107.6 ± 8.69 1.8 . (j, ('() 48 207.3 ± 22.02 13.8 410.3 ± 32.60 17.8 268.1 ± 27.51 13.9 65.2 ± 8.08 2.8 91.4 ± 8.61 2.1 l"l ;:s- a 72 107.3 ± 17.82 7.3 322.9 ± 26.13 18.1 181.3 ± 25.42 11.1 67.7 ± 2.46 8.0 60.9 ± 9.96 3.7 ~ Fl· :- :I ~ .... ~CYS +MET ~EAK ~AK l::l. "' , hou ('() ;:.;- mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) 2 519.5 ± 38.29 35.2 6507.0 ± 131.54 40.5 12442.2 ± 137.94 25.0 12 332.0 ± 10.55 7.1 4888.8 ± 153.32 28.1 9516.8 ± 113.24 27.0 24 191.9 ± 6.98 7.7 3021.7 ± 147.72 10.9 6212.7 ± 171.15 13.4 36 183.4 ± 7.48 9.1 3046.6 ± 137.59 10.1 6204.2 ± 115.38 15.0 48 156.6 ± 12.97 8.5 2731.8 ± 132.29 10.8 5597.4 ± 138.71 15.5 72 128.7 ± 12.99 10.5 1842.3 ± 55.22 6.7 3996.1 ± 54.74 13.6 IV: Dynamics o change s in con en o non -essen ial amino acids in colos um o dai y cows on he 2nd and subsequen lac a ions wi hin he i s 2-72 h a e pa u i ion (mg /JOO ml) ASP SER GLU PRO hou mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) 2802.9 ± 35.62 9.9 677.4 ± 15.11 5.0 2252.1 ± 311.62 30.9 991.0 ± 22.31 5.0 12 598.1 ± 50.77 18.9 534.5 ± 25.51 10.7 1860.9 ± 177.88 21.4 695.6 ± 45.82 14.7 24 427.8 ± 30.42 15.9 366.0 ± 20.52 12.5 1262.6 ± 171.40 30.4 488.5 ± 29.63 13.5 36 433.9 ± 44.71 23.1 363.4 ± 41.18 25.3 1183.8 ± 255.27 48.2 494.0 ± 42.78 19.3 48 395.7 ± 27.68 15.6 312.5 ± 29.17 20.8 1078.7 ± 199.65 41.4 470.1 ± 50.59 24.1 72 296.6 ± 30.44 22.9 231.5 ± 31.19 30.0 850.1 ± 32.10 8.4 387.4 ± 30.02 17.5 GLY ALA TYR LNEAK hou mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) 2 284.8 ± 35.74 28.1 374.4 ± 22.43 13.4 552.6 ± 32.61 13.2 5935.2 ± 104.71 10.3 12 227.3 ± 22.63 22.3 281.7 ± 25.11 19.9 430.0 ± 51.68 26.8 4628.0 ± 83.25 8.5 24 154.4 ± 26.01 37.7 203.4 ± 36.33 40.5 288.3 ± 30.62 23.7 3191.0± 27.96 19.6 36 154.1 ± 18.63 27.1 219.9 ± 23.81 24.2 308.5 ± 26.38 19.1 3 157.6± 30.65 12.7 48 138.0 ± 33.91 41.8 191.2 ± 8.03 9.7 279.3 ± 21.19 16.9 2865.5 ± 31.56 14.6 72 81.2 ± 13.25 10.3 130.6 ± 16.63 8.5 176.6 ± 11.52 6.0 2153.7 ± 10.98 11.4 00 -..l Y:Dynamics o changes in con en o essen ial amino acids in cow scolos um wi hin he ji s 2- 72h a e pa u i ion (mg /IOO ml) 00 00 THR YAL ILE LEU PHE hou mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) 2760.3 ± 22.14 19.2 681.1 ± 14.74 16.8 663.8 ± 25.62 12.2 11 48.3 ± 19.61 15.4 611.7 ± 19.26 10.0 12 579.3 ± 53.45 29.2 676.5 ± 45.61 21.3 459.7 ± 42.52 29.3 785.5 ± 36.76 14.8 448.6 ± 31.61 22.3 24 380.7 ± 54.86 24.8 454.9 ± 79.35 25.9 315.5 ± 39.43 19.4 546.2 ± 79.46 46.0 294.6 ± 27.43 17.9 36 359.3 ± 81.39 22.7 434.1 ± 55.88 12.9 303.7 ± 53.74 29.6 531.0 ± 82.38 26.1 279.5 ± 29.28 33.1 48 334.9 ± 35.41 10.6 393.4 ± 73.41 15.9 284.2 ± 55.51 18.9 518.7 ± 83.22 27.8 279.2 ± 23.82 28.5 VJ 72 213.8 ± 26.26 12.3 288.4 ± 81.05 12.3 182.1 ± 24.47 13.4 387.4 ± 102.14 26.4 173.6 ± 26.35 15.2 ~ 1::> . ~ ' I::> ..... HIS LYS ARG CYS MET ~ hou o mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) ::: ::s , ;.;- 2 462.9 ± 22.40 15.3 998.9 ± 32.53 10.3 635.7 ± 54.40 27.0 300.2 ± 37.67 39.7 233.3 ± 31.85 43.2 . 1::> ~ 12 356.7 ± 26.77 23.7 708.2 ± 57.60 25.7 458.6 ± 31.14 23.5 163.9 ± 29.44 56.7 164.9 ± 36.09 40.1 ~ 24 248.1 ± 31.37 12.6 468.0 ± 46.55 28.8 306.7 ± 37.96 28.2 93.2 ± 16.46 25.6 115.4 ± 27.84 24.0 .1::> 36 235.0 ± 25.53 23.4 441.8 ± 21.97 15.7 300.3 ± 27.81 18.8 74.9 ± 20.06 27.5 105.0 ± 32.72 10.2 ~ . 48 231.0 ± 25.43 21.1 427.5 ± 19.73 16.3 285.9 ± 20.68 17.2 73.6 ± 28.96 12.4 88.8 ± 33.01 10.4 (J , ( ) (') 72 134.2 ± 28.88 21.5 346.3 ± 25.09 17.3 201.8 ± 22.23 11.0 59.3 ± 28.89 15.0 61.9 ± 38.12 6.2 ::s- o ..:: . 1::>. :- l LCYS+MET LEAK LAK ~ -- hou 1::>. ,,>, mean±S.D. CY(%) mean±S.D. CY(%) mean±S.D. CY(%) ( ) ;.;- 2533.5 ± 61.67 36.6 6496.2 ±88.52 43.0 12445.2 ± 83.33 21.0 12 328.9 ± 55.67 53.5 4802.0 ± 95.42 20.1 9418.2 ± 134.12 19.5 24 208.6 ± 56.96 28.6 3223.4 ± 67.66 16.6 6672.5 ± 154.29 17.3 36 179.9 ± 40.39 22.5 3064.5 ± 58.29 10.1 6131.8 ± 103.45 16.9 48 162.4 ± 67.11 21.2 2917.2 ± 62.49 6.8 5876.5 ± 95.48 15.1 72 121.3 ± 78.22 17.3 2048.8 ± 43.66 11.4 4222.5 ± 80.88 16.1 VI: Dynamics o changes in con en o non -essen ial amino acids in cows colos um wi hin he i s 2 -72 h c ie pa u i ion (mg /IOO ml) ASP SER OLD PRO hou CV(%) mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. 2806.3 ± 44.39 17.4 679.7 ± 24.04 11.2 2255.2 ± 292.08 14.I 997.6 ± 42.06 13.3 12 598.5 ± 30.85 16.3 534.8 ± 19.58 11.6 1852.2 ± 93.28 15.9 694.7 ± 26.79 12.2 24 465.0 ± 86.56 18.5 408.1 ± 17.06 13.2 1301.0 ± 129.74 31.5 522.0 ± 41.34 16.8 36 421.7 ± 48.94 16.7 361.3 ± 22.83 20.0 1143.1 ± 186.70 51.6 487.4 ± 34.29 22.2 48 409.3 ± 49.15 13.8 330.9 ± 20.65 16.2 1073.9 ± 120.02 35.3 483.6 ± 55.76 26.5 72 290.4 ± 49.80 13.4 225.9 ± 25.99 13.6 875.8 ± 134.74 38.2 381.5 ± 27.01 22.4 GLY ALA TYR LNEAK hou mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) mean±S.D. CV(%) 2290.2 ± 26.58 29.0 368.4 ± 26.51 22.8 551.7 ± 20.83 11.9 5949.0 ± 82.04 14.4 12 228.3 ± 18.22 25.2 276.6 ± 23.67 27.1 431.0 ± 16.86 19.7 4616.2 ± 108.70 17.4 24 193.7 ± 13.22 21.6 234.9 ± 25.16 14.6 324.4 ± 22.16 11 .8 3449.1 ± 87.12 17.9 36 142.6 ± 13.43 19.4 211.8 ± 10.54 14.9 299.3 ± 35.05 27.1 3067.3 ± 107.93 25.2 48 157.8 ± 22.48 14.3 208.6 ± 18.81 19.0 295.2 ± 54.35 28.2 2959.3 ± 114.32 26.9 72 88.3 ± 32.02 11.5 . 132.5 ± 16.22 13.9 179.3 ± 25.86 24.6 2173.7 ± 142.62 20.7 o :: § 0C1 (J) C/l S' 3 S' o ~ 0: C/l (") o 3 '0 o ~. .... o' :::3 o ...., n ~ C/l o o 0' C/l 2' 3 ,-., 0- s:: ..., S' 0C1 ::n ..., C/l .... -..J IV :: o s:: ..., C/l ~ ~ ..., '0 ~ ::!• .... o' :::3 '-"