Reserch Article Turk. J. Vet. Anim. Sci. 2010; 34(3): 255-260 TÜBİTAK doi:10.3906/vet-0811-2 Blood plsm minerl profile nd helth sttus in postprtum cows fed n nionic diet efore prturition Czim CRNKIC 1, *, Slko MURATOVIC 2, Slvic PIPLICA 1, Aid KAVAZOVIC 1, Stnimir KUTLACA 3 1 Deprtment of Animl Nutrition, Fculty of Veterinry Medicine, University of Srjevo, Zmj od Bosne 90, 71000 Srjevo - BOSNIA & HERZEGOVINA 2 Animl Science Institute, Fculty of Agriculture nd Food Science, University of Srjevo, Zmj od Bosne 8, 71000 Srjevo - BOSNIA & HERZEGOVINA 3 PD Butmir Diry Frm, Doglodi, 71210 Ilidz - BOSNIA & HERZEGOVINA Received: 06.11.2008 Astrct: The effects of n nionic versus ctionic (control) diet on plsm minerl levels nd helth sttus were investigted in 24 dry pregnnt Holstein-Friesin cows during the lst 3 weeks of pregnncy. The cows were in their third or fourth prity, hd moderte level of milk production in their previous lcttion, nd did not hve history of milk fever. The nionic diet significntly improved plsm clcium sttus, s compred to the control (2.18 vs. 1.99 mmol L 1 ) nd nonsignificntly reduced the incidence of suclinicl hypoclcemi (1/12 vs. 5/12) without ffecting other periprturient diseses. Other plsm minerl levels were unffected, except chloride, which ws higher in the nionic group. The results show tht n nionic diet could improve clcium sttus round the time of prturition nd reduce the incidence of suclinicl hypoclcemi when fed to moderte producing diry cows not predisposed to milk fever. Key words: Anionic diet, dry cow, helth sttus, plsm minerls Introduction An nionic, or cidic, diet is one supplemented with nionic slts to provide more nions (Cl nd S 2 ) reltive to the ctions N + nd K +. It hs negtive dietry ction-nion difference (DCAD), clculted s meq kg 1 of dry mtter = (N + K) (Cl + S). It is well known tht nionic diets efficiently mintin lood C t prturition nd prevent milk fever when fed to cows during the lst severl weeks of pregnncy (1-4). Milk fever (presis puerperlis) my hve serious consequences for susequent milk production (1), ut some investigtions suggest tht ttention should lso e pid to suclinicl hypoclcemi (5,6). Reducing smooth muscle contrctions, nd rumen nd intestinl motility (7), suclinicl hypoclcemi my hve n even greter impct on herd economy thn clinicl milk fever ecuse of its roder influence on dry mtter intke (DMI), secondry disese conditions, nd milk production during erly lcttion (8). Feeding n nionic diet efore prturition hs een dvised if the incidence of milk fever in herd * E-mil: czim.crnkic@vfs.uns. 255
Blood plsm minerl profile nd helth sttus in postprtum cows fed n nionic diet efore prturition exceeds 10%, nd lso when it is desired to improve the helth sttus nd production in herds in which clinicl milk fever is not serious prolem (8,9). Milk fever occurs more frequently in high producing cows (10) nd in cows with history of the disese (1). Diry herds in Bosni nd Herzegovin (B&H) generlly do not hve prolems with milk fever nd the incidence of the disese seems to e low. Some field experience suggests n incidence of out 2% in herds, ut more thn 40% of cows my hve suclinicl hypoclcemi fter prturition (V. Skic, personl communiction). It is lso importnt to know tht the level of milk production is moderte in most diry herds in B&H. It remins unknown if use of nionic slts in moderte producing herds cn e useful. The im of the present study ws to exmine the effects of n nionic diet fed efore prturition on C sttus nd other plsm minerl levels fter clving, s well s on the helth sttus in moderte producing cows without history of milk fever. Mterils nd methods The study included 24 pregnnt non-lctting Holstein-Friesin cows without history of milk fever tht were 65.5 ± 5 months old (men ± SD), weighed 660 ± 32 kg, nd in their third or fourth prity. The cows were rndomly llocted to 2 groups ccording to prity, expected clving dte, nd milk yield during the previous lcttion (6574 ± 681 kg) (Tle 1), which were offered 1 of the 2 experimentl diets (control nd nionic) during the lst 21 dys of pregnncy. The experiment ws conducted t Butmir Tle 1. Bseline nlysis of the cows (men ± SD) fed nionic or control diets in the preprtum period. Control Anionic P = group group Age, months 65.7 ± 5.9 65.4 ± 4.2 0.907 Prity 3.4 ± 0.5 3.6 ± 0.5 0.436 Milk production, kg 6557 ± 719 6592 ± 671 0.901 Body weight, kg 657 ± 33 661 ± 31 0.767 Dys on preprtum diet 19.1 ± 5.5 18.3 ± 3.9 0.705 In previous lcttion. Diry Frm, Srjevo city region, B&H. Cows were housed nd fed in tie stll with drinking wter ville d liitum. Feeding of nionic slts ended t prturition. Immeditely fter clving, cows entered the routine nutrition nd mngement progrm used t Butmir Diry Frm. The diets consisted of corn silge with grss hy nd dditionl mounts of sunflower mel, whet rn, nd commercil feed mixture, s well s slt nd minerl-vitmin supplement (Tle 2). All the concentrtes nd supplements were mixed efore feeding nd fed s concentrte mixture. Pre-clving DMI ws set t 10.1 kg cow 1 dy 1 in oth groups (1.53% of ody weight) nd contined out 75% dry mtter (DM) from roughge. The cows were fed 5%- 10% less thn the expected voluntry intke to ensure consumption of the desired mount of feed without refusl. The nionic diet contined n nionic slt mixture tht included (s fed) 140 g of MgSO 4 7H 2 O, 80 g of CCl 2 2H 2 O, nd 20 g of NH 4 Cl (totl: 2.6 Eq) for ech niml ech dy. The slts were mixed with the concentrte mixture nd offered twice dily t 0800 nd 1500, following portion of grss hy nd efore corn silge. Tle 2. Ingredient composition (DM, %) of preprtum diets used in n experiment to determine the effects of preprtum nionic diet on plsm minerl nd helth sttus fter clving. Control diet Anionic diet Grss hy 30.19 30.20 Corn silge 45.53 45.55 Whet rn 3.46 3.47 Sunflower mel 9.21 9.21 Feed mixture 10.89 9.51 Min-vit. supplement 0.38 0.38 NCl 0.20 0.20 CCl 2 2H 2 O - 0.60 MgSO 4 7H 2 O 0.14 0.68 NH 4 Cl - 0.20 Supplemented per 1 kg with: vitmin A 10,000 IU, vitmin D 3 1500 IU, vitmin E 20 mg, J 1 mg, Mn 50 mg, Co 0.12 mg, Se 0.1 mg, Fe 25 mg, nd Zn 45 mg. Contined per 1 kg: C 160 g, P 80 g, N 100 g, Mg 20 g, Cl 154 g, vitmin A 1,000,000 IU, vitmin D 3 120,000 IU, vitmin E 1200 mg, vitmin B 1 60 mg, vitmin B 2 30 mg, vitmin B 6 15 mg, vitmin B 12 100 μg, nicin 600 mg, Zn 9000 mg, Fe 800 mg, Mn 5000 mg, Cu 1000 mg, Co 20 mg, J 100 mg, nd Se 40 mg. 256
C. CRNKIC, S. MURATOVIC, S. PIPLICA, A. KAVAZOVIC, S. KUTLACA Feed smples were nlyzed for DM, crude protein, fier, nd ether extrct, s well s C, P, Mg, N, K, Cl, nd S. Asolute DM content of ech feed ws determined y drying t 103-105 C for 24 h. Dried smples were ground through 1-mm screen nd shed t 550 C overnight. Nitrogen content ws determined using Kjeltec Model 1030 utomtic nlyzer (Tector AB, Hognos, Sweden). Crude fier nlysis nd ft extrction were performed ccording to stndrd ISO procedures. C, Mg, N, nd K concentrtions were determined vi tomic sorption spectrophotometry (AAnlyst 300, Perkin Elmer Corp., Norwlk, CT, USA) in shed feed smples dissolved in 3N HCl solution nd diluted (1:50) either with strontium chloride for C nd Mg or with deionized wter for N nd K nlysis. Phosphorus ws mesured colorimetriclly using the mmonium molydte procedure. Sulfur ws mesured using the grvimetric method, with mgnesium nitrte nd precipittion with rium chloride. Chlorine ws mesured using sodium cronte nd precipittion with silver nitrte. Anlysis of C nd Mg in nionic slts ws performed in wter solution using the sme procedure s for the feeds, while S nd Cl were mesured using the grvimetric method, with rium chloride precipittion, nd the titrimetric method of Mohr, respectively. Chemicl nlysis of minerlvitmin supplements ws not performed, except for DM nd sh content; therefore the reported composition of the product ws used for clcultions. Whole lood smples were collected neroiclly from the jugulr vein of ech cow into 10-mL heprinized vcutiners (Becton Dickinson VACUTAINER Systems, Rutherford, NJ, USA). All lood smples were otined within 8 h of prturition, immeditely plced on ice, nd trnsported to the l, where they were centrifuged t 800 g for 20 min. After centrifugtion plsm ws trnsferred to plstic tues nd frozen t 20 C until minerl element nlysis. The entire procedure (from lood smple collection to plsm seprtion) ws performed within 2 h of smpling. Prior to nlysis, plsm smples were thwed t room temperture overnight nd the minerl concentrtions were determined the next dy. Totl C, Mg, N, nd K concentrtions were nlyzed vi tomic sorption spectrophotometry. Smples were diluted 1:50 with 0.1% (wt.:vol.) lnthnum-chloride for C nd Mg, nd with deionized wter for N nd K nlysis. Inorgnic plsm P ws mesured colorimetriclly with mmonium chloride fter smples were deproteinized with 20% trichlorocetic cid. Plsm Cl ws mesured using the method of Mohr. All clinicl diseses were dignosed nd treted y the frm s veterinry service, nd hypoclcemi (plsm C <2 mmol L 1 ) nd hypomgnesemi (plsm Mg <0.75 mmol L 1 ) were dignosed y lortory nlysis of lood plsm smples. Student s t-test for independent smples ws used to determine the significnce of the differences etween corresponding mens in the 2 groups. Differences in disese incidences were tested y the 2-tiled Fisher's exct test. P vlues less thn or equl to 0.05 were considered sttisticlly significnt. Results Diet composition nd intke The nutrient composition of the preprtum diets clculted fter chemicl nlysis of the feed ingredients is shown in Tle 3. The diets were similr Tle 3. Clculted nutrient composition of preprtum diets (DM sis) used in n experiment to determine the effects of preprtum nionic diet on plsm minerl nd helth sttus fter clving. Control diet Anionic diet Crude protein, % 12.02 12.06 Crude fier, % 26.39 26.33 Ether extrct, % 3.42 3.37 Ash, % 5.69 6.74 NEL, MJ kg 1 5.60 5.49 Clcium, % 0.55 0.76 Phosphorus, % 0.40 0.39 Mgnesium, % 0.26 0.36 Sodium, % 0.14 0.14 Potssium, % 1.01 1.01 Chlorine, % 0.25 0.76 Sulfur, % 0.20 0.34 DCAD, meq kg 1 + 127.32 107.43 Net energy for lcttion ws sed on tulr vlues for ll feeds (18), ut clculted for the commercil feed mixture. Clculted s (N + K) (Cl + S). 257
Blood plsm minerl profile nd helth sttus in postprtum cows fed n nionic diet efore prturition with respect to ll nutrients nd energy, except for S nd Cl. Levels of C nd Mg were out 40% higher in the nionic diet ecuse of the high levels of these minerls in nionic slts. The experimentl feeding period ws scheduled to lst 21 dys efore prturition, ut, ecuse of norml vrition in the predicted clving dtes, cows in the control nd nionic groups spent 19.1 ± 5.5 nd 18.3 ± 3.9 dys on the tretment (men ± SD), respectively (Tle 1). During the tretment period cows in oth groups consumed the diets completely; the only oserved difference ws tht cows in the nionic group took severl minutes longer to consume the sme mount of concentrte feed thn the cows in the control group (dt not shown). Plsm minerls The minerl concentrtions in lood plsm re shown in Tle 4. Totl plsm C ws significntly higher t clving in the cows fed the nionic diet thn in those fed the control diet. Slightly higher levels of inorgnic P nd lower levels of plsm Mg were oserved in the nionic group, ut the differences etween the groups were not significnt. Concentrtions of plsm N nd K did not differ etween the groups, ut the plsm Cl concentrtion ws significntly higher in the nionic group. The strong ion difference (SID) in lood plsm ws lower in the cows fed the nionic diet (s ws expected), ut this ws not significntly different compred to tht in the cows fed the control diet. Tle 4. Postprturient plsm minerl concentrtions (men ± SD) in cows fed nionic or control diets in the preprtum period. Mmol L 1 Control Anionic P = group group Clcium 1.99 ± 0.22 2.18 ± 0.23 0.049 Phosphorus 1.36 ± 0.45 1.48 ± 0.43 0.506 Mgnesium 1.06 ± 0.17 0.95 ± 0.16 0.112 Sodium 136.83 ± 6.38 138.28 ± 8.75 0.649 Potssium 4.53 ± 0.40 4.56 ± 0.61 0.891 Chlorine 98.47 ± 4.37 103.83 ± 4.70 0.008 SID, meq L 1 42.88 ± 6.46 38.98 ± 9.17 0.242 Inorgnic. Strong-ion difference clculted s (N + + K + ) Cl. Helth sttus The helth sttus of the cows fter prturition is shown in Tle 5. Five cows in the control group hd suclinicl hypoclcemi fter prturition, while only 1 cse ws oserved in the nionic group; the difference ws not significnt (P = 0.155). Two cows hd plsm Mg concentrtions elow 0.75 mmol L 1 (1 in ech group). There were no differences in the incidence of retined plcent, metritis, or mstitis etween the groups. Three complex cses (cows experiencing 2 or more clinicl diseses fter prturition) were seen in the control group, while none were oserved in the nionic group. Ech of the complex cses ws hypoclcemic. Discussion The use of nionic slts is esier if totl mixed rtion (TMR) is used, ut it is lso possile to use them in component feeding systems if the slts re mixed with t lest 2.3 kg of concentrte to msk their itter tste (11). In B&H roughge nd concentrtes re lwys fed seprtely, nd TMRs generlly hve not een used in the field. Oservtions mde during the present study suggest tht the tste of the slts ws Tle 5. Incidence of postprtum helth disorders in cows fed nionic or control diets during the preprtum period. Control Anionic Fisher s, group group P = Suclinicl hypoclcemi 5/12 1/12 0.155 Hypomgnesemi c 1/12 1/12 1.000 Retined plcent d 2/12 1/12 1.000 Metritis 3/12 2/12 1.000 Mstitis 2/12 2/12 1.000 Complex cses e 3/12 0/12 0.217 Totl helth disorders f 7/36 5/36 0.753 c d e f Numer of cses per numer of cows in ech group. Suclinicl hypoclcemi defined s <2 mmol L 1 of totl C in lood plsm. Hypomgnesemi defined s <0.75 mmol L 1 of totl Mg in lood plsm. Fetl memrnes retined longer thn 12 h fter prturition. Numer of cows tht experienced 2 or more clinicl disorders. Suclinicl hypoclcemi nd hypomgnesemi not included in this clcultion. 258
C. CRNKIC, S. MURATOVIC, S. PIPLICA, A. KAVAZOVIC, S. KUTLACA not completely msked in 2.6 kg of concentrte feed, ut the mount of concentrte ws sufficient to ensure consumption of the desired mount of nionic slts. The results of this study gree with those of numerous previous works tht hve reported positive effect of nionic diets on C sttus in periprtum cows (1,2,4,12-14). The reltively smll difference in plsm C etween the nionic nd control groups, lthough significnt, ws not sufficient to cuse significnt chnges in plsm P nd Mg. Phillippo et l. (15) reported tht chnges in P nd Mg metolism round the time of prturition re relted to hypoclcemi, rther thn to the diet. Hypoclcemi releses more prthyroid hormone, which increses resorption of C nd Mg in renl tuules nd inhiits resorption of P t the sme time (16). As consequence, reduction in the plsm C concentrtion t prturition is usully ccompnied y hypophosphtemi nd hypomgnesemi (15). However, this is not lwys the cse, especilly if hypoclcemi is not severe nd is not cliniclly mnifested. These findings correspond to the results of Oetzel et l. (12) nd Joyce et l. (17), who lso did not oserve ny chnges in plsm P nd Mg concentrtions on the dy of clving, despite the fct tht cows fed nionic diets hd significntly higher level of plsm C thn cows fed ctionic diets. The oserved chnges in plsm N, K, nd Cl in the present study re in ccordnce with the dietry tretment, s oserved y others (15). Most of the cows in the present study were 5 or more yers old, which is considered criticl ge for milk fever, ut clinicl cses of the disese were not pprent. The proility of the incidence of the disese ws not high, considering tht no cow chosen for this experiment experienced milk fever during previous clving (1) nd milk yield during the previous lcttion ws not high (10). The positive effect of the nionic diet on suclinicl hypoclcemi prevention (1/12 vs. 5/12) ws in ccordnce with erlier reports relted to high producing cows (2,6,12). The difference in the incidence of hypoclcemi ws numericlly importnt, ut not significnt (P = 0.155), most certinly ecuse of the smll smple size. One cow in the control group nd nother in the nionic group hd plsm Mg concentrtions elow 0.75 mmol L 1. The norml concentrtion of plsm Mg in cows rnges from 0.75 to 1 mmol L 1 nd levels elow 0.75 mmol L 1 suggest insufficient intke or poor iovilility of Mg in the digestive trct (18). Dietry Mg intke ws sufficient in the control group nd even incresed in the nionic group, ut this hd no effect on plsm Mg t clving. Incresing Mg intke in cows fed nionic slts from 0.2% to 0.37% lso yielded no enefit with regrd to C or Mg metolism in previous experiment (19). Oetzel et l. (12) reported tht cows fed nionic diets re less likely to develop disorders relted to hypoclcemi, nd recent experiments showed tht those diets fed during lte gesttion significntly reduced the incidence of retined plcent postprtum (14,20). However, nionic diets did not ffect the incidence of helth disorders in other studies (2,17), which is in ccordnce with the present study s findings, which might hve een relted to the reltively good C sttus in oth groups of cows. There were 5 hypoclcemic cows in the control group, ut plsm C vlues never fell elow 1.70 mmol L 1, which is still considered mild hypoclcemi. All the complex cses (cows tht experienced 2 or more clinicl diseses) oserved in the present study were relted to hypoclcemic cows in the control group. The cow with the lowest plsm C concentrtion hd retined plcent, followed y coliform mstitis with serious generl helth disturnces nd complete cesstion of milk secretion. The other 2 cows hd retined plcent, plus metritis nd metritis plus mstitis. This finding confirms tht periprturient disorders re often seen s complex nd re interrelted (10). The prolem ssocited with the investigtion nd interprettion of results on helth sttus often lies in the reltively low numer of nimls used in such studies. All ut 2 (2,20) of the studies cited in this pper were conducted with smll smples. The numer of cows per group in the present study ws lso too smll to relily detect differences in disese incidences; therefore cution must e tken when interpreting the results. In conclusion, the nionic diet fed efore prturition in the present study resulted in tendency 259
Blood plsm minerl profile nd helth sttus in postprtum cows fed n nionic diet efore prturition towrds improved C sttus nd decrese in the incidence of suclinicl hypoclcemi in moderte producing diry cows tht were not predisposed to milk fever. Acknowledgement The uthors thnk Mr. Scir Sivro for his finncil support, nd Lutvo Memic nd Mirsd Memisevic for their technicl ssistnce. References 1. Block, E.: Mnipulting dietry nions nd ctions for preprtum diry cows to reduce incidence of milk fever. J. Diry Sci., 1984; 67: 2939-2948. 2. Beede, D.K., Wng, C., Donovn, G.A., Archild, L.F., Snchez, W.K.: Dietry ction-nion difference (electrolyte lnce) in lte pregnncy. Proceedings of Florid Diry Production Conference, Ginesville, FL., 1991; 98-104. 3. Horst, R.L., Goff, J.P., Reinhrdt, T.A.: Adpting to the trnsition etween gesttion nd lcttion: differences etween rt, humn nd diry cow. J. Mmmry Glnd Biol., 2005; 10: 141-156. 4. Kuroski, N., Ymto, O., Mori, F., Imoto, S., Mede, Y.: Preventive effect of mildly ltering dietry ction-nion difference on milk fever in diry cows. J. Vet. Med. Sci., 2007; 69: 185-192. 5. Goff, J.P., Horst, R.L., Jrdon, P.W., Borelli, C., Wedm, J.: Field trils of n orl clcium propionte pste s n id to prevent milk fever in periprturient diry cows. J. Diry Sci., 1996; 79: 378-383. 6. Cozzi, G., Dl Coroo, F., Pesvento, A., Rvrotto, L., Gi, G.: Effects of two different preprtum diets on some metolic trits nd productive response in multiprous Holstein cows in erly lcttion. Itl. J. Anim. Sci., 2005; 4: 127-137. 7. Rmerg, C.F. Jr., Johnson, E.K., Frgo, R.D., Kronfeld, D.S.: Clcium homeostsis in cows, with specil reference to prturient hypoclcemi. Am. J. Physiol. Regul. Integr. Comp. Physiol., 1984; 246: 698-704. 8. Horst, R.L., Goff, J.P., Reinhrdt, T.A.: Clcium nd vitmin D metolism in the diry cow. J. Diry Sci., 1994; 77: 1936-1951. 9. Oetzel, G.R.: Use of nionic slts for prevention milk fever in diry cttle. Comp. Cont. Educ. Prct., 1993; 15: 1138-1146. 10. Gröhn, Y.T., Er, H.N., McCulloch, C.E., Sloniemi, H.S.: Epidemiology of metolic disorders in diry cttle: ssocition mong host chrcteristics, disese, nd production. J. Diry Sci., 1989; 72: 1876-1885. 11. Oetzel, G.R., Brmore, J.A.: Intke of concentrte mixture contining vrious nionic slts fed to pregnnt, nonlctting diry cows. J. Diry Sci., 1993; 76: 1617-1623. 12. Oetzel, G.R., Olson, J.D., Curtis, C.R., Fettmn, M.J.: Ammonium chloride nd mmonium sulphte for prevention of prturient presis in diry cows. J. Diry Sci., 1988; 71: 3302-3309. 13. Goff, J.P., Horst, R.L.: DCAD ffects responsiveness of cows to prthyroid hormone; how nion supplementtion prevents milk fever. Americn Diry Science Assocition Astrcts, 2004; 280. 14. Wu, W.X., Liu, J.X., Xu, G.Z., Ye, J.A.: Clcium homeostsis, cid se lnce, nd helth sttus in periprturient Holstein cows fed diets with low ction nion difference. Livest. Sci., 2008; 117: 7-14. 15. Phillippo, M., Reid, G.W., Nevison, I.M.: Prturient hypoclcemi in diry cows: effects of dietry cidity on plsm minerls nd clciotrophic hormones. Res. Vet. Sci., 1994; 56: 303-309. 16. Littledike, E.T., Goff, J.P.: Interctions of clcium, phosphorus, mgnesium nd vitmin D tht influence their sttus in domestic met nimls. J. Anim. Sci., 1987; 65: 1727-1743. 17. Joyce, P.W., Snchez, W.K., Goff, J.P.: Effect of nionic slts in preprtum diets sed on lflf. J. Diry Sci., 1997; 80: 2866-2875. 18. Ntionl Reserch Council: Nutrient Requirement of Diry Cttle. 7th rev. edn., Ntionl Acdemy Press, Wshington, DC., 2001; 128-131. 19. Wng, C., Beede, D.K.: Effects of diet mgnesium on cid-se sttus nd clcium metolism of dry cows fed cidogenic slts. J. Diry Sci., 1992; 75: 829-836. 20. Melendez, P., Donovn, G.A., Risco, C.A., Goff, J.P.: Plsm minerl nd energy metolite concentrtions in diry cows fed n nionic preprtum diet tht did or did not hve retined fetl memrnes fter prturition. Am. J. Vet. Res., 2004; 65: 1071-1076. 260