NEUTROPHIL AND LYMPHOCYTE RESPONSE TO VITAMINS C AND E SUPPLEMENTATION IN YOUNG CALVES. S. D. Pruiett, J. L. Morrill, F.

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1 NEUTROPHIL AND LYMPHOCYTE RESPONSE TO VITAMINS C AND E SUPPLEMENTATION IN YOUNG CALVES 1 S. D. Pruiett, J. L. Morrill, F. Blech, J. J. Higgins, N. V. Anderson, nd P. G. Reddy Summry Clves were ottle-fed milk replcers t 10% of weekly djusted ody weight for 8 wk. Tretments were 1) no supplements (control), 2).16 oz vitmin C, or 3).16 oz vitmin C plus 125 IU/l vitmin E. Lymphocytes nd neutrophils isolted from dy 14 nd dy 28 lood smples were ssyed for neutrophil-medited S. ureus phgocytosis nd ntiody-dependent cellulr cytotoxicity, nd for mitogen induced lymphocyte prolifertion. Eye nd nsl dischrges of clves supplemented with vitmin C nd vitmins C plus E were less thn those of control clves for wk 1 to 8. Lymphocyte prolifertion with the mitogens showed trend for higher responses t wk 2 in vitmin C plus E supplemented clves. Neutrophils of clves supplemented with vitmin C showed decresed phgocytosis nd lysis functions compred to those of control clves t wk 2 nd 4. Neutrophil function of clves supplemented with vitmins C plus E ws ner or slightly higher thn tht of controls t wk 2 nd 4, suggesting tht the ddition of vitmin E negted the dverse effects tht vitmin C lone hd on neutrophil functions. Introduction Ascoric cid (vitmin C) is produced y the liver of mny nimls, including cttle. Ascoric cid synthesis egins in clves etween the second nd third wk of life nd reches dult concentrtions of vitmin C round 3 mo of ge. Milk, which hs reltively low scoric cid content, is often exposed to ir nd light efore eing consumed y the clf. Both re destructive gents of vitmin C. Therefore, vitmin C deficiency is potentil prolem with the young milk-fed clf. This would e especilly true for diry clves, whose only source of vitmin C for the first few wk of life is ucketor ottle-fed milk or milk replcer. Vitmin C deficiency hs een linked to decresed immune response, nd elevted scoric cid concentrtions hve een linked to incresed immune response in ctfish, poultry, cttle, nd swine. Conversely, others hve found tht vitmin C supplementtion hd no eneficil effect on the immune responses mesured. The vlue of vitmin E to the immune system of the young clf hs een estlished. It is primrily responsile for protection of the cell memrne. In other species, vitmins C nd E hve een shown to work coopertively to protect the cell memrnes ginst peroxidtion. The purpose of the present study ws to determine the effects of supplements of vitmin C lone nd vitmins C nd E together on the function of neutrophils nd lymphocytes of the young clf's 1 Deprtment of Antomy nd Physiology. 2 Deprtment of Sttistics. 3 Deprtment of Surgery nd Medicine. 35

2 immune system, s well s their effect on the growth nd generl helth of the clf during the first 8 wk of life. Procedures Thirty Holstein ull clves were removed from their dms t 24 h following irth. They were given trnsition milk for 2 more dys, then ssigned to ech of three tretment groups. The tretments consisted of milk replcers 1) without supplementl vitmins (control), 2) with.16 oz vitmin C per kg, or 3) with.16 oz vitmin C plus 125 IU vitmin E per l. Milk replcer ws reconstituted to 13.5% dry mtter nd ws fed t 10% of ody weight, djusted weekly, which ws divided into two equl dily feedings. Clves were housed in individul hutches with strw edding. Wter ws ville d liitum. Body weights were mesured nd milk replcer lloctions were djusted weekly. Twice dily fecl nd generl ppernce scores were recorded, nd eye or nsl dischrge or signs of enteric or respirtory illness were noted. Blood smples were collected in heprinized tues on experimentl dy 1, 14, 28, nd 56 for vitmin C nd E determintions. Dy 14 nd dy 28 smples were used to ssy lymphocyte prolifertion nd neutrophil function. A lymphocyte trnsformtion ssy (LTA) nd neutrophil-medited ntiody-dependent cellulr-cytotoxicity (ADCC) nd S. ureus phgocytosis ssys were used to determine lymphocyte nd neutrophil cellulr function. Results nd Discussion Vitmin C ws stle in the milk replcers fter decrese tht occurred prior to smpling. Plsm vitmin C concentrtions of the control group dropped throughout the study until wk 8, when concentrtions for ll tretments were pproximtely equl (Figure 1). The vitmin C-only group mintined high concentrtion until wk 8. The plsm vitmin C content of vitmin C plus E- supplemented clves decresed gretly t wk 2, ut lter recovered to levels ner those of the vitmin C-only group. Plsm vitmin E concentrtions reflected the supplementtion with tht vitmin. No significnt differences occurred etween tretments in gin or feed efficiency until wk 6 (Tle 1). The vitmin C plus E supplemented group hd greter gins over the entire 8 wk period thn the vitmin C supplemented group nd ws more efficient in feed conversion (l feed/l gin) thn oth the control nd vitmin C supplemented group. Tle 2 shows weekly men fecl scores nd dischrge oservtions for ll groups. The control group tended to hve the lowest fecl scores (more solid feces) throughout the 8 wk, nd the vitmin C supplemented group tended to hve the highest scores. The control nd vitmin C supplemented groups were different (P<.10) t wk 2, 6, nd 8, nd t wk 8 the vitmin C group ws lso significntly higher thn the vitmin C plus E supplemented group. Men eye nd nose dischrges tended to e higher for the control group for ll 8 wk. At wk 3, the vitmin C plus E supplemented clves hd significntly less dischrges thn the control group, nd the vitmin C only supplemented group ws significntly lower thn the control clves t wk 7. 36

3 Tle 1. Totl nd Dily Gin y Week nd Feed Efficiency Gin, l/d Feed efficiency, l feed/l gin Vitmin supplementtion Vitmin supplementtion Wk None C C + E None C C + E Totls Differing superscripts within the sme week denote mens with sttisticlly significnt differences (P<.10). Becuse of vriility in the lymphocyte functions of the 2- nd 4-wk-old clf, no significnt differences etween the tretment groups could e noted in the Con-A nd PHA mitogen-induced prolifertion mens. Week 2 lymphocytes of vitmin C plus vitmin E supplemented clves showed trends towrd greter prolifertion. By wk 4, the lymphocytes of vitmin C supplemented clves hd prolifertive responses equl to or greter thn those of the C plus E supplemented clves. The lck of differences in lymphocyte prolifertion etween tretments my hve een prtly due to very little environmentl stress experienced y these clves. Environmentl stress hs een shown to increse the use of vitmin C nd ntiody production in young clves supplemented with vitmin C. Incresed ntiody production would e preceded y incresed lymphocyte prolifertion. The neutrophil medited ADCC t wk 2 (Figure 2) showed significnt differences etween the two vitmin supplemented groups, nd t wk 4 etween the vitmin C supplemented group nd the control s well. The S. ureus phgocytosis ssys showed the vitmin C group to e significntly different from the control t wk 2, ut no significnt differences were detected t wk 4. 37

4 Tle 2. Weekly Eye nd Nsl Dischrge nd Weekly Fecl Scores Differing superscripts differences (P<.10). within the sme week denote mens with sttisticlly significnt Conclusion Supplementtion with vitmin C lone nd vitmins C nd E together did not hve eneficil effect on ny of the immune responses mesured here. However, the lower incidence of mucous dischrge from eyes nd noses suggests eneficil effect tht ws not reflected y the cellulr functions we mesured. The use of vitmins C nd E together ppered to negte the dverse effects of vitmin C lone on neutrophil functions t oth 2 nd 4 wk nd on lymphocyte prolifertion t wk 2. Figure 1. Plsm vitmin C concentrtions t 0, 2, 4, nd 8 wk of tretment. 38

5 Figure 2. Neutrophil-medited ntiody-dependent cellulr cytotoxicity % specific-lysis nd S. ureus phgocytosis % kill. Differing superscripts within the sme wk denote mens cd with significnt differences (P<. 10). Differing superscripts within the sme wk denote mens with significnt differences (P <.05). 39