Proeedings, Western Setion, Amerin Soiety of Animl Siene #89 Vol. 66, 015 Meloxim s n Alterntive to Allevite Inflmmtory nd Aute-Phse Retions in Beef Cttle Upon Lipopolyshride Administrtion or Vintion M. C. Rodrigues 1,, R. F. Cooke 1, R. S. Mrques 1, S. Arispe, D. H. Keisler, nd D. W. Bohnert 1 Oregon Stte University - Estern Oregon Agriulturl Reserh Center, Burns, OR 1 UNESP - Fuldde de Mediin Veterinári e Zooteni, Botutu, Brzil Oregon Stte University Mlheur County Extension, Ontrio, OR University of Missouri - Division of Animl Sienes, Columi, MO Astrt text: Twenty-one Angus steers (n = 11) nd heifers (n = 10) were ssigned to Exp. 1 (d -1 to 6) nd Exp. (d 6 to 0). On d -10, ttle were housed in individul pens nd offered free-hoie wter, minerl-vitmin mix, nd hy until d 0. In Exp. 1, lves were rnked on d -1 y gender nd BW, nd ssigned to: 1) orl meloxim dministrtion (1 mg/kg of BW dily) from dy -1 to 6 (MEL7), ) orl meloxim dministrtion (1 mg/kg of BW) on d 0, nd orl ltose monohydrte dministrtion (1 mg/kg of BW) on d -1 nd from d 1 to 6 (MEL1), nd ) orl ltose monohydrte dministrtion (1 mg/kg of BW dily) from d -1 to 6 (CON). On d 0, ll ttle reeived n intrvenous lipopolyshride olus (0.5 μg/kg of BW). From d - to d 6, ttle BW nd hy DMI were reorded dily. Retl temperture ws ssessed nd lood smples olleted every h from - to 8 h, every 6 h from 1 to 7 h, nd every h from 96 to 1 h reltive to lipopolyshride dministrtion. Clves reeiving MEL7 hd greter (P = 0.0) hy DMI (kg/d) ompred with MEL1 nd CON. When DMI ws evluted s % of BW, MEL7 hd greter (P = 0.05) hy DMI ompred with CON. No tretment effets were deteted (P = 0.90) for retl temperture, plsm ortisol, insulin, leptin, hptogloin nd serum NEFA, lthough temperture nd plsm vriles inresed fter LPS dministrtion (time effets; P < 0.01). For Exp., lves were mintined nd reeived the sme tretments s in Exp. 1 from d 6 to 1. On d 7, ttle were vinted ginst respirtory pthogens. Cttle BW ws reorded t the eginning (d 6 nd 7) nd end (d 0 nd 1) of Exp., wheres hy DMI ws reorded dily. Retl temperture ws ssessed nd lood smples olleted s in Exp. 1, in ddition to 168, 0, nd 6 h reltive to vintion. No tretment effets were deteted (P 0.15), lthough hy DMI deresed, plsm onentrtions of ortisol, insulin, leptin, nd hptogloin inresed, nd serum ntiodies ginst respirtory pthogens lso inresed (time effets, P < 0.01) fter vintion. Hene, meloxim filed to modulte the physiologil, inflmmtory, nd ute-phse protein retions ssoited with lipopolyshride dministrtion nd vintion in eef ttle. Keywords: ttle, lipopolyshride, meloxim, vintion INTRODUCTION Orl meloxim dministrtion to feeder ttle llevited the ute-phse protein retion nd prevented the derese in reeiving ADG, DMI, nd G:F used y trnsport nd feedlot entry (Gurnieri Filho et l., 01), inditing tht meloxim is n lterntive to mitigte inflmmtory retions nd performne losses eliited y stressful events. Nevertheless, reserh is still required to further understnd the role of meloxim during utephse nd inflmmtory retions (Vn Engen et l., 01) to iologilly support its enefits to highly-stressed eef ttle. One lterntive to hrterize the effets of meloxim on the ovine innte immune system is through teril lipopolyshride (LPS) hllenge nd susequent evlution of physiologil nd ute-phse vriles (Crroll et l., 009). Vintion is nother stressful nd mndtory proedure in ttle opertions (Crroll nd Forserg, 007). As n exmple, vintion ginst Mnnheimi hemolyti, the min terium isolted from ovine respirtory disese (BRD) in feedlot ttle (Rie et l., 007), stimulted n ute-phse protein retion nd redued ADG, DMI, nd G:F during the wk susequent to vintion (Arthington et l., 01; Mrques et l., 01). Hene, reserh to develop mngement interventions tht enefit vine-indued immune protetion nd ttle performne is wrrnted (Arthington et l., 01). Bsed on the enefits of meloxim dministrtion to highlystressed ttle (Gurnieri Filho et l., 01; Vn Engen et l., 01) it n e hypothesized tht orl meloxim will llevite the ute-phse response nd prevent the derese in performne used y vintion. Bsed on this rtionle, the ojetive of this experiment ws to evlute the effets of orl meloxim dministrtion on performne, inflmmtory, nd utephse prmeters of eef ttle ssigned to teril LPS hllenge (Exp. 1), or vinted ginst the BRD omplex (Exp. ). MATERIALS AND METHODS Twenty-one Angus hlter-trined steers (n = 11) nd heifers (n = 10) were ssigned to the Exp. 1 (d -1 to 6) nd susequently to Exp. (d 6 to 0). Cttle were wened t 6 mo of ge on d -, nd exposed dily to hlter-trining tehniques until d -1. At wening, ttle were vinted 67
ginst lostridil diseses, infetious ovine rhinotrheitis, ovine virl dirrhe omplex, pneumoni, nd M. hemolyti (One Shot Ultr 7, Bovi-Shield Gold One Shot, nd TSV-; Zoetis, Florhm Prk, NJ), nd dministered n nthelminti (Detomx; Zoetis). On d -10, ttle were housed in individul pens ontined within n enlosed rn, nd offered free hoie wter, minerlvitmin mix, nd mixed lflf-grss hy until the end of Exp. (d 0). Experiment 1 Animls nd tretments. On d -1, lves were rnked y gender nd BW (vg. BW = ± kg), nd ssigned to 1 of tretments: 1) orl meloxim dministrtion (1 mg/kg of BW dily; Crlsd Tehnologies, In., Crlsd, CA) from dy -1 to 6 (MEL7), ) orl meloxim dministrtion (1 mg/kg of BW; Crlsd Tehnologies, In.) on d 0, nd orl ltose monohydrte dministrtion (1 mg/kg of BW, exipient used in the mnufture of meloxim tlets; Avntor Performne Mterils, Center Vlley, PA) on d -1 nd from d 1 to 6 (MEL1), nd ) orl ltose monohydrte dministrtion (1 mg/kg of BW dily; Avntor Performne Mterils) from d -1 to 6 (CON). Meloxim ws originlly presented in 15 mg tlets, whih were ground dily using ommeril food proessor (Soho Food Proessor; West Bend Housewres, West Bend, WI) to ensure tht ttle reeived their ext dose. Ltose monohydrte ws dministered to ount for potentil pleo effets. Meloxim or ltose monohydrte were mnully mixed with 50 ml of 0.9% sline nd dministered individully to ttle vi orl drenh. The MEL7 nd CON tretments re sed on Gurnieri Filho et l. (01), whih lso suggested tht different lengths meloxim dministrtion should e investigted. Aordingly, the MEL1 tretment ws inluded to determine if single meloxim dministrtion n mitigte the inflmmtory nd ute-phse responses eliited y the LPS hllenge, whih our within h hllenge (Crroll et l., 009). On d 0, ll ttle reeived n intrvenous olus dose of teril LPS (0.5 μg/kg of BW, Esherihi oli 0111:B, Sigm-Aldrih, St. Louis, MO) onurrently with tretment dministrtion (Crroll et l., 009). Bteril LPS ws dissolved into 10 ml of 0.9% sline immeditely efore dministrtion. Smpling. From d - to d 6, ttle were weighed dily. Hy DMI ws evluted dily from d - to 6 from eh pen y olleting nd weighing refusls dily. Smples of the offered nd non-onsumed feed were olleted dily from eh pen nd dried for 96 h t 50 C in fored-ir ovens for DM lultion. Steer retl temperture ws ssessed with GLA M750 digitl thermometer (GLA Agriulturl Eletronis, Sn Luis Oispo, CA) every h from - to 8 h, every 6 h from 1 to 7 h, nd every h from 96 to 1 h reltive to LPS dministrtion. Blood smples were olleted onurrently with retl temperture ssessment vi jugulr venipunture into ommeril lood olletion tues (Vutiner, 10 ml; Beton Dikinson, Frnklin Lkes, NJ) with or without freeze-dried sodium heprin for plsm nd serum olletion, respetively. Blood smples were pled immeditely on ie, entrifuged (,500 g for 0 min; C) for plsm or serum hrvest, nd stored t -80 C on the sme dy of olletion. All plsm smples were nlyzed for plsm hptogloin onentrtions (Gurnieri Filho et l., 01). Smples olleted from - to 8 h reltive to LPS dministrtion were nlyzed for onentrtions of serum NEFA, plsm ortisol, insulin, nd leptin (Delvud et l., 000; Gurnieri Filho et l., 01). Experiment Animls nd tretments. Immeditely fter the lst smpling of Exp. 1 on d 6, ttle (vg. BW = 8 ± kg) were ssigned to the sme tretment sheme reeived in Exp. 1. Tretments (MEL7, MEL1, nd CON) were dministered from d 6 to d 1. On d 7, ttle were re-vinted ginst infetious ovine rhinotrheitis, ovine virl dirrhe omplex, pneumoni, nd M. hemolyti (Bovi-Shield Gold One Shot; Zoetis) onurrently with tretment dministrtion. As in Exp. 1, the MEL7 nd CON tretments re sed on Gurnieri Filho et l. (01). Given tht leukoytes responsile for inflmmtory nd ute-phse responses re diretly involved with ntigen presenttion to T ells (Durum nd Muegge. 1996), exessive meloxim dministrtion my impir the innte immune responses required for proper vine effiy. Therefore, MEL1 ws inluded to determine if single meloxim dministrtion onurrently with hndling for vintion n mitigte the resultnt inflmmtory nd ute-phse responses (Mrques et l., 01) without impiring vine effiy. Smpling. Cttle full BW ws reorded t the eginning (d 6 nd 7) nd end (d 0 nd 1) of the experiment. Hy DMI ws reorded dily from d 5 to 0 s in Exp. 1, wheres G:F ws lulted sed on totl BW gin nd hy DMI during the experimentl period. Retl temperture ws ssessed nd lood smples olleted s in Exp. 1, with dditionl olleted t 168, 0, nd 6 h reltive to vintion for nlysis of plsm hptogloin (Gurnieri Filho et l., 01). Plsm smples olleted immeditely prior (0 h) nd t 168, 0, nd 6 h following vintion were lso nlyzed for onentrtions of ntiodies ginst M. hemolyti (Confer et l., 009), s well s titers ginst ovine respirtory synytil virus (BRSV), ovine herpesvirus-1 (BHV-1), ovine virl dirrhe virus-1 (BVD-1), nd prinfluenz virus (PI) t the Oklhom Animl Disese Dignosti Lortory (Stillwter, OK). Sttistil nlysis Dt from oth experiments were nlyzed using the MIXED proedure of SAS (SAS Inst., In., Cry, NC) nd Stterthwite pproximtion to determine the denomintor df for the tests of fixed effets. Animl ws onsidered the experimentl unit. The model sttements ontined the effets of tretment, time, gender, nd ll resultnt intertion. Hy DMI ws nlyzed using vlues otined on d - nd -1 (Exp. 1) or d 5 nd 6 (Exp. ) s ovrites. Animl(tretment gender) ws used s rndom vrile. The speified term for the repeted sttement ws time, niml(tretment gender) ws inluded s sujet, nd the ovrine struture utilized ws utoregressive, whih provided the lowest Akike informtion riterion nd 68
therefore the est fit. Results re reported s lest squre mens, ovritely djusted mens for hy DMI, nd seprted using PDIFF. Signifine ws set t P 0.05, nd tendenies were determined if P > 0.05 nd 0.10. Tle 1. Performne nd retl temperture (RT) of steers nd heifers hllenged with lipopolyshride (LPS; Exp. 1) or vintion (Exp. ), nd ssigned to reeive orl meloxim (1 mg/kg of BW dily) for 7 d (MEL7), ltose monohydrte for 7 d (CON), or meloxim for 1 d nd ltose monohydrte for 6 d (MEL1) during the hllenging period. 1 Exp. 1 BW, kg 9 1 1 7 0.61 DMI Kg/d.7 5.5.75 0. 0.05 % of BW 1.97..10 0.1 0.09 RT 8.99 8.95 8.96 0.07 0.90 Exp. BW, kg Initil 0 7 0.8 Finl 8 8 7 0.55 BW hnge.98.98 5. 1.0 0.80 DMI Kg/d 5.50 5.7 5.0 0.19 0.8 % of BW.8.7.5 0.08 0.55 G:F, g/kg 71. 5.5 80. 1.0 0.66 RT 9.19 9.06 9.1 0.06 0.9 1 In Exp. 1, ll ttle reeived n intrvenous olus dose of teril LPS (0.5 μg/kg of BW, Esherihi oli 0111:B, Sigm-Aldrih, St. Louis, MO). In Exp., ttle were vinted ginst infetious ovine rhinotrheitis, ovine virl dirrhe omplex, pneumoni, nd Mnnheimi. hemolyti (Bovi-Shield Gold One Shot; Zoetis, Florhm Prk, NJ). RESULTS AND DISCUSSION Experiment 1 No tretment effets were deteted (P = 0.61) for lf BW during the experimentl period (Tle 1). Clves ssigned to MEL7 hd greter (P = 0.0) tul hy DMI ompred with MEL1 nd CON lves (Tle 1). When DMI ws evluted s % of BW, MEL7 hd greter (P = 0.05) hy DMI ompred with CON lves only (Tle 1), wheres hy DMI of MEL1 lves ws similr (P 0.5) ompred with MEL7 nd CON ohorts (Tle 1). Therefore, these results indite tht orl meloxim dministrtion for 7 d llevited the derese in hy DMI used y LPS dministrtion nd the stress of hndling for smple olletion (dy effet, P < 0.01; Figure 1). No tretment effets were deteted (P = 0.90) for retl temperture, whih inresed for ll tretments (time effet, P < 0.01; dt not shown) from to 6 h nd returned to seline levels 8 h reltive to LPS dministrtion. No tretment effets were deteted (P 0.7) for plsm ortisol, insulin, leptin, hptogloin nd serum NEFA (Tle ). Time effets were deteted (P < 0.01; dt not shown) for ll plsm vriles. Plsm ortisol onentrtions inresed (P < 0.05) from to 6 h nd returned to seline levels 8 h reltive to LPS dministrtion. Plsm insulin onentrtions inresed (P < 0.05) from to 1 h nd returned to seline levels 16 h reltive to LPS dministrtion. Plsm leptin onentrtions inresed (P < 0.05) from 8 to 16 h nd returned to seline levels h reltive to LPS dministrtion. Plsm hptogloin onentrtions inresed (P < 0.05) eginning t 16 h nd returned to seline levels only t 1 h reltive to LPS dministrtion. Crroll et l. (009) reported similr time effets for retl temperture nd plsm ortisol in wened steers reeiving LPS t 1.0 μg/kg of BW. The time effet deteted for hptogloin demonstrtes tht LPS dministrtion effetively indued n ute-phse protein retion (Crroll et l., 009). Time effets deteted for plsm insulin nd leptin indite hnges in energy metolism to ope with the stress of inflmmtion (Wggoner et l., 009). Nevertheless, ll these vriles were similr mong CON, MEL7, nd MEL1, suggesting tht orl meloxim dministrtion filed to modulte the physiologil nd inflmmtory responses triggered y LPS dministrtion. Tle. Blood vriles from steers nd heifers hllenged with lipopolyshride (LPS; Exp. 1) or vintion (Exp. ), nd ssigned to reeive orl meloxim (1 mg/kg of BW dily) for 7 d (MEL7), ltose monohydrte for 7 d (CON), or meloxim for 1 d nd ltose monohydrte for 6 d (MEL1) during the hllenging period. 1 Exp. 1 Cortisol, ng/ml 0. 0.6 0.1.7 0.98 Insulin, µiu/ml.7.0.75 0.66 0.9 NEFA, μeq/l 0. 0. 0. 0.05 0.98 Leptin, ng/ml.89.9 5. 0. 0.7 Hptogloin, ng/ml 0.6 0.9 0.8 0.05 0.95 Exp. Cortisol, ng/ml 6.6 6.1. 1.6 0.61 Insulin, µiu/ml 1.8 1.9 1.91 0.8 0.98 NEFA, μeq/l 0.7 0.9 0.5 0.06 0.68 Leptin, ng/ml 5.00 5.9 5. 0.5 0.6 Hptogloin, ng/ml 0.57 0.5 0.56 0.09 0.9 1 In Exp. 1, ll ttle reeived n intrvenous olus dose of teril LPS (0.5 μg/kg of BW, Esherihi oli 0111:B, Sigm-Aldrih, St. Louis, MO). In Exp., ttle were vinted ginst infetious ovine rhinotrheitis, ovine virl dirrhe omplex, pneumoni, nd Mnnheimi hemolyti (Bovi-Shield Gold One Shot; Zoetis, Florhm Prk, NJ). Experiment No tretment effets deteted (P = 0.80) BW hnge during the experimentl period (Tle 1). No tretment effets were deteted (P 0.8; Tle 1) for hy DMI prmeters, lthough DMI deresed (dy effet, P < 0.01; Figure ) upon vintion s previously reported y Mrques et l. (01). No tretment effets were deteted (P = 0.9) for retl temperture, whih inresed for ll tretments (time effet, P < 0.01; dt not shown) from to 16 h nd returned to seline levels h reltive to vintion. No tretment effets were deteted (P 0.61) for plsm ortisol, insulin, leptin, hptogloin nd serum NEFA 69
(Tle ). Time effets were deteted (P < 0.01; dt not shown) for ll plsm vriles. Plsm ortisol onentrtions inresed (P < 0.05) from to 1 h nd returned to seline levels 16 h reltive to vintion. Plsm insulin onentrtions inresed (P < 0.05) from 8 to 16 h nd returned to seline levels h reltive to vintion. Plsm leptin onentrtions inresed (P < 0.05) from 8 to 16 h nd returned to seline levels h reltive to vintion. Plsm hptogloin onentrtions inresed (P < 0.05) eginning t 16 h nd returned to seline levels only t 168 h reltive to vintion. Similr outomes were reported y Mrques et l. (01), illustrting the physiologil hnges, inluding energy metolism, inflmmtory nd stress retion, used y vintion ginst BRD pthogens in eef ttle. Tle. Serum onentrtions of ntiodies ginst M. hemolyti (ng ound), s well s serum titers (log) ginst ovine respirtory synytil virus (BRSV), ovine herpesvirus-1 (BHV-1), ovine virl dirrhe virus-1 (BVD-1), nd prinfluenz virus (PI) in steers nd heifers hllenged with vine ginst these pthogens nd ssigned to reeive orl meloxim (1 mg/kg of BW dily) for 7 d (MEL7), ltose monohydrte for 7 d (CON), or meloxim for 1 d nd ltose monohydrte for 6 d (MEL1) during the hllenging period. 1 M. hemolyti 0.87 0.70 0.96 0.17 0.5 BRSV 1.70 1.57 1. 0.16 0.6 BHV-1 0.79 0.66 0.75 0. 0.90 BVD-1 1.5 1.5 1.17 0.1 0.6 PI 1.65 1.01 1.5 0. 0.16 1 Cttle were vinted ginst infetious ovine rhinotrheitis, ovine virl dirrhe omplex, pneumoni, nd Mnnheimi hemolyti (Bovi- Shield Gold One Shot; Zoetis, Florhm Prk, NJ). No tretment effets were deteted (P 0.16) for serum onentrtions of ntiodies ginst M. hemolyti, or serum titers ginst BRSV, BHV-1, BVD-1, nd BVD-1. Time effets were deteted (P < 0.01) for ll titers ut for BVD (P = 0.0), given tht vlues inresed (P 0.0) when ompring smples olleted prior to nd fter vintion (0.68 vs. 0.89, SEM = 0.05 for M. hemolyti, 1.0 vs. 1.70, SEM = 0.11 for BRSV, 0. vs. 0.90, SEM = 0.15 for BHV-1, nd 0.81 vs. 1.50, SEM = 0.1 for PI. These results indite tht vintion indued the expeted ntiody response ut for BVD-1, wheres meloxim dministrtion did not impt these outomes. IMPLICATIONS Orl meloxim dministrtion to eef ttle for 7 onseutive d t 1 mg/kg of BW prevented the derese in DMI used y LPS dministrtion, ut did not llevite the resultnt inflmmtory nd ute-phse protein retions. Similrly, orl meloxim dministrtion to eef ttle t 1 mg/kg of BW upon vintion ginst the BRD omplex did not prevent the vine-indued derese in DMI, did not llevite the resultnt inflmmtory nd ute-phse protein responses, nd did not impt serum onentrtions of ntiodies ginst these pthogens. Hene, meloxim dministrtion filed to modulte the physiologil hllenges ssoited with LPS dministrtion nd vintion in eef ttle. LITERATURE CITED Arthington, J. D., R. F. Cooke, T. D. Mddok, D. B. Arujo, P. Moriel, N. DiLorenzo, nd G. C. Lm. 01. Effets of vintion on the ute-phse protein response nd mesures of performne in growing eef lves. J. Anim. Si. 91:181-187. Crroll, J. A., nd N. E. Forserg. 007. Influene of stress nd nutrition on ttle immunity. Vet. Clin. Food. Anim. :105-19. Crroll, J. A., Reuter, R.R., Chse Jr., C.C., Colemn, S.W., Riley, D.G., Spiers, D.E., Arthington, J.D. nd Glyen, M.L. 009. Profile of the ovine ute-phse response following n intrvenous lipopolyshride hllenge. Innte Immun. 15:81 89. Confer, A. W., S. Aylew, M. Montelongo, D. L. Step, J. H. Wry, R. D. Hnsen, nd R. J. Pnier. 009. Immunity of ttle following vintion with Mnnheimi hemolyti himeri PlpE LKT (SAC89) protein. Vine 7:1771 1776. Delvud, C., F. Boquier, Y. Chillird, D. H. Keisler, A. Gertler, nd G. Knn. 000. Plsm leptin determintion in ruminnts: Effet of nutritionl sttus nd ody ftness on plsm leptin onentrtion ssessed y speifi RIA in sheep. J. Endorinol. 165:519 56. Gurnieri Filho, T. A., R. F. Cooke, B. I. Cppellozz, M. M. Reis, R. S. Mrques, nd D. W. Bohnert. 01. Effets of meloxim dministrtion on physiologil nd performne responses of trnsported feeder ttle. J. Anim. Si. 9:17-1. Mrques, R. S., R. F. Cooke, B. I. Cppellozz, T. Gurnieri Filho, M. M. Reis, D. H. Keisler, nd D. W. Bohnert. 01. A vine-indued ute-phse retion inreses plsm leptin onentrtions in eef ttle. J. Anim. Si. 9(E-Suppl. ):58. Rie, J. A., L. Crrso-Medin, D. C. Hodgins, nd P. E. Shewen. 007. Mnnheimi hemolyti nd ovine respirtory disese. Anim. Helth Res. Rev. 8:117 18. Vn Engen, N. K., M. L. Stok, T. Engelken, R. C. Vnn, L. W. Wulf, L. A. Krriker, W. D. Busy, J. Lkritz, A. J. Crpenter, B. J. Brdford, W. H. Hsu, C. Wng, nd J. F. Coetzee. 01. Impt of orl meloxim on irulting physiologil iomrkers of stress nd inflmmtion in eef steers fter long-distne trnsporttion. J. Anim. Si. 9:98 510. Wggoner, J. W., C. A. Löest, J. L. Turner, C. P. Mthis, nd D. M. Hllford. 009. Effets of dietry protein nd teril lipopolyshride infusion on nitrogen metolism nd hormonl responses of growing eef steers. J. Anim. Si. 009. 87:656 668. 70
Figure 1. Hy DMI, expressed s kg/d (1A) or % of BW (1B), of steers nd heifers dministered intrvenous teril liposhride (LPS; 0.5 μg/kg of BW, Esherihi oli 0111:B, Sigm-Aldrih, St. Louis, MO) on d 0 of the experiment. A dy effet ws deteted (P < 0.01) for oth vriles. Vlues with different letters differ t P < 0.05. Figure. Hy DMI, expressed s kg/d (A) or % of BW (B), of steers nd heifers vinted ginst infetious ovine rhinotrheitis, ovine virl dirrhe omplex, pneumoni, nd Mnnheimi hemolyti (Bovi-Shield Gold One Shot; Zoetis, Florhm Prk, NJ) on d 7 of the experiment. A dy effet ws deteted (P < 0.01) for oth vriles. Vlues with different letters differ t P < 0.05. 7 6 5 - -1 0 1 5 6 (1A) 8 6 0 5 6 7 8 9 10 11 1 1 1 15 16 17 18 19 0 d d d d d d d d d d (A) d d 1 (1B) - -1 0 1 5 6 Dy reltive to LPS dministrtion (d 0) 1 0 (B) 5 6 7 8 9 10 11 1 1 1 15 16 17 18 19 0 Dy reltive to vintion (d 7) 71