HAEMATOLOGICAL AND BIOCHEMICAL FINDINGS IN PREGNANT, POSTFOALING AND LACTATING JENNIES

Transcription

1 REVISED HAEMATOLOGICAL AND BIOCHEMICAL FINDINGS IN PREGNANT, POSTFOALING AND LACTATING JENNIES Bonelli F 1, Rot A 1, Corzz M 1, Serio D 1, Sgorini M 1 1 Deprtment of Veterinry Sciences, vi Livornese snc, 56122, Sn Piero Grdo (PI), Itly. Corresponding uthor Dr Micel Sgorini Deprtment of Veterinry Sciences, vi Livornese snc, 56122, Sn Piero Grdo (PI), Itly. Phone numer: +39(0) Fx numer: +39(0) Emil: micel.sgorini@unipi.it 1

2 Astrct The ims of this study were to: 1) verify if significnt chnges occur in hemtologicl nd iochemicl prmeters in jennies during the lst two months of pregnncy nd the first two months of lcttion, nd 2) determine ny differences with equine species. Mterils nd methods. Hemtologicl nd iochemicl prmeters were evluted in jennies every 15 dys during lte pregnncy, prturition, nd erly lcttion. The Kolmogorov-Smirnov test, ANOVA for repeted mesurements nd Tukey s multiple comprison test s post hoc were pplied. The significnce level ws set t p<0.05. Results. Sttisticl nlysis showed differences relted to time for RBC nd HCT, WBC, PLT, totl proteins (TP), lood ure, triglycerides nd totl cholesterol concentrtions, AST, GGT, CK ctivities, sodium (N) nd potssium (K). Discussion nd conclusions. RBC nd HCT were higher in lte pregnncy thn t foling nd during lcttion. The reltive nemi might e due to incresed wter ingestion due to fluid losses. The WBC count ws higher t foling thn during lte pregnncy nd lcttion. This could e relted to the relese of cortisol nd ctecholmine during delivery. The PLT trend showed lower vlues from delivery to the first two months of lcttion compred to lte gesttion. Blood ure incresed ner prturition, nd then remined constnt during delivery nd lcttion, which might e due to the high-energy demnd t the eginning of lcttion. Triglycerides nd totl cholesterol showed decrese from delivery through the lcttion period. Thus jennies seem to hve similr metolism of fts to ponies nd drft horse mres, chrcterized y greter ft content nd moiliztion thn light reed horses. AST ctivity decresed t prturition nd erly lcttion, proly due to predominnce of nolic over ctolic processes during pregnncy. GGT ctivity ws lower t delivery nd during lcttion thn t lte gesttion. This could e due to physiologicl lod on the liver in the perintl period. GGT ctivity ws lwys higher thn in mres, ut within the norml rnge for dult donkeys. CK decresed ner delivery, then ws constnt from prturition through the first two months of lcttion. N decresed during lcttion, proly due to n incresed renl retention medited y ldosterone relese during pregnncy. K showed the sme trend s N, nd concentrtions re in line with the species. The higher K during pregnncy my e due to resorption y the gut. TP decresed more during the post-prtum period nd lcttion thn in the gesttionl period. 2

3 Key words Jennies, hemtology, iochemistry, pregnncy, foling, lcttion. 3

4 Introduction Donkeys (Equus sinus) hve een close compnions to humns for millenni nd hve een used s working nimls ll over the world. Donkey milk could possily e used in children with intolernce to cow s milk [1,2] or in niml-ssisted therpy [3]. The renewed interest in these nimls is demonstrted y the numer of studies on estlishing the se-line dt of oth hemtologicl nd iochemicl vriles in the lood of dult [4-26], s well s in neworn donkeys [27-28]. Pregnncy nd lcttion re physiologicl periods tht result in incresed metolic demnds. Although homeosttic mechnisms keep sustnces in the lood t reltively constnt levels, some chnges in the concentrtions of routine clinicl chemistry nlytes re likely to occur [29]. During pregnncy, n expnsion in plsm volume nd erythrocyte mss occurs, s well s n increse in plsm protein synthesis [30]. In women, it is well known tht during pregnncy reference intervls re different from the non-pregnnt stte [31-33]. The effects of pregnncy, prturition nd lcttion on hemtologicl prmeters hve only een studied in horses to limited extent [34-44] in different reeds. Moreover, only one study [42] hs reported thorough investigtion of the lst months of pregnncy nd first month of lcttion nd no dt re present in literture concerning hemtologicl nd iochemicl vlues in jennies during the sme period. Therefore, more detiled prmeters in hemtology nd iochemistry during lte pregnncy, prturition nd lcttion in jennies could e useful for ccurte dignosis of diseses. The ims of this study were to: 1) verify if significnt chnges occur in hemtologicl nd iochemicl prmeters in jennies during the lst two months of pregnncy nd the first two months of lcttion; nd 2) determine ny differences with respect to equine species 2. Mteril nd Methods 2.1. Animls The study ws conducted on 9 jennies elonging to the Amit donkey reed for totl of 18 pregnncies nd 16 fols orn during two-yer study. Jennies were from 5 to 13 yers old, weighed 300 to 350 kg, nd were kept in collective pddocks t the Veterinry Teching Hospitl, Deprtment of Veterinry Sciences, Pis University. Approvl for this study ws otined from the Ethicl Committee on Animl Experimenttion of the University of Pis, nd the protocol ws sent to the Ministry of Helth. 4

5 In order to provide NRC recommendtions for energy, jennies were fed with medow hy d liitum long with commercil equine feed This feeding procedure egn t two months pre-prtum, nd continued through post-prtum nd the first two months of lcttion. Jennies were housed together during pregnncy in pddock 10x20 m. Close to prturition jennies were housed in 4x4 m oxes for the first 15 dys of lcttion, nd then returned to the pddock. Jennies were included in this study ccording to the following criteri: 1) pregnncy length 353.4±13.0 dys [43]; 2) unssisted delivery; 3) tretment ginst gstrointestinl prsites nd vccinted ginst equine influenz, tetnus, nd equine herpes virus-1, in ccordnce with the guidelines of the Americn Assocition of Equine Prctitioners Infectious Disese Committee [44] Smple collection nd hndling Blood smples were otined from the jugulr vein. Ech jenny ws smpled every 15 dys during lte pregnncy, pproximtely two months efore prturition, t prturition, nd every 15 dys during the first two months of lcttion. Blood ws collected in test tues contining K2-EDTA (cod , FL Medicl, Pdu, Itly) nd lithium-heprin test tues (cod , FL Medicl, Pdu, Itly). To void ltertions relted to diurnl vritions, lood smples were collected t the sme time ech dy (8:00-9:00 m), except for the smple collected t prturition Complete lood count K2-EDTA smples were nlysed with cell counter (Hecovet C /ITA, nd CAL- SEAC multiprmetric hemtology clirtor, SEAC-RADIM Co, Florence, Itly) t lest 5 minutes fter the collection. The im ws to determine: 1) erythrocyte count (RBC), 2) leukocyte count (WBC), 3) hemogloin concentrtion (Hg), 4) men corpusculr volume (MCV), 5) men corpusculr hemogloin (MCH), 6) men corpusculr hemogloin concentrtion (MCHC), 7) pltelet count (PLT). Specimens contining clots or grossly hemolysed were excluded. A qulity control of the cell counter ws performed ech dy Biochemicl nlysis 5

6 Heprinised smples were centrifuged t 3000 g for 10 minutes, s recommended y the mnufcturer. Plsm ws then frozen t -18 C nd nlysed within 15 dys fter collection. Clinicl chemistry ws performed with n utonlyzer (Lisys, Anlyzer Medicl System-AMS, Rome, Itly; qulity control norml level: ASR02010 nd pthologic level: ASR02020, Assel Srl, Rome, Itly). The prmeters nlysed were: 1) glucose concentrtion (Glucose SL, enzymtic colorimetric method, cod. ASR01202, Assel Srl, Rome, Itly); 2) cretinine (Cretinine, kinetic modified Jffè method, cod. ASR01150, Assel Srl, Rome, Itly); 3) lood ure (Ure UV SL, kinetic enzymtic method, cod. ASR01143, Assel Srl, Rome, Itly); 4) triglycerides (Triglycerides-SL, enzymtic colorimetric method, cod. ASR01134, Assel Srl, Rome, Itly); 5) totl cholesterol (Cholesterol liquid, trinder method-endpoint, cod. 7050, FAR, Veron, Itly); 6) totl iliruin (Totl Biliruin, colorimetric method without DMSO, cod. ASR01034/1, Assel Srl, Rome, Itly); 7) sprtte minotrnsferse (AST) (AST SL, kinetic method UV -IFCC- cod. ASR01220, Assel Srl, Rome, Itly); 8) gmm glutmyl trnsferse (γgt) (Gmm GT SL, kinetic method-szsz-tris, cod. ASR01194, Assel Srl, Rome, Itly); 9) cretine-phosphokinse (CK) (CK NAC SL, kinetic method UV, cod. ASR01074, Assel Srl, Rome, Itly); 10) lkline phosphtse (ALP) (Alkline Phosphtse SL-DGKC- kinetic method, cod. ASR01162, Assel Srl, Rome, Itly); 11) totl clcium (Tot- C) (Clcium OCPC, colorimetric method, cod. ASR01050, Assel Srl, Rome, Itly); 12) potssium (K) (phox Plus L, Stt Profile, Nov Biochemicl, Miln, Itly); 13) sodium (N) (phox Plus L, Stt Profile, Nov Biochemicl, Miln, Itly); 14) phosphorus (P) (Phosphorus UV, direct method with molidte, cod , Sec-Rdim, Florence, Itly); 15) totl protein (TP); 16) lumin (Alumin BCG, romocresol green method, cod , Sec-Rdim, Florence, Itly). The sme opertor lwys performed the iochemistry profile, ccording to stndrd methods Sttisticl nlysis Averge (X) nd stndrd devition (SD) vlues were clculted for ech hemtologicl nd iochemicl prmeter collected t ll smpling times, long with minimum nd mximum vlues nd the 95% of confidence intervl (CI). The Kolmogorov-Smirnov test ws crried out to verify dt distriution. Results showed n pproximtely Gussin distriution, thus nlysis of vrince for repeted mesurements nd Tukey s multiple comprison test s post hoc were pplied. The significnce level ws set t p<0.05. All nlyses were performed using commercil softwre (GrphPd Prism, USA). 6

7 Results None of the smples collected contined clots or were grossly hemolysed. The results for hemtologicl nd iochemicl prmeters expressed s men ± stndrd devition re reported in Tles 1 nd 2, respectively. Not ll the jennies were smpled t ech time point, thus the ctul numers of smples for ech time point re indicted in Tles 1 nd 2, respectively. In prticulr, 1 out of 2 jenny died 1 week fter delivery due to colic surgery, nd 1 out of 2 ws excluded fter delivery ecuse her fol died due to septicemi. Sttisticl nlysis showed differences relted to smpling times for RBC nd HCT, WBC, PLT, totl protein, lood ure, triglycerides nd totl cholesterol concentrtions, AST, GGT, CK ctivities, P, N, nd K. 7

8 Discussion nd Conclusions All niml species need specific reference intervls of hemtologicl nd iochemicl prmeters for good interprettion of lood smple nlyses. The im of the present pper ws to study hemtologicl nd iochemicl profiles in jennies during lte pregnncy, post-prtum period nd lcttion. Jennies enrolled in this study hd wide ge rnge (5-13 yers) ecuse ge seems not to influence hemtologicl nd iochemicl prmeters [42,44]. The RBC nd HCT trend showed higher vlues in lte pregnncy thn foling time nd lctting period. Thus, jennies did not seem to e ffected y nemi in lte gesttion, s in mres [42], ut not s women [46] nd other niml species [47-49]. The reltive nemi (lower HCT) during lcttion might e relted to lrge increse in wter ingestion secondry to fluid losses with the eginning of lcttion nd susequent over-hydrtion nd erythrocyte dilution, s lredy reported for mres [42]. Compred to vlues in mres, the results in jennies were lower throughout the study period [37-39,42]. The trend in WBC count showed higher vlues t foling thn lte pregnncy nd lcttion, in greement with previous studies on mres [37,42]. This could e relted to the relese of cortisol nd ctecholmine during delivery or to neutrophil mrgintion into the uterus in the post-prtum period [42,50]. In terms of WBC vlues, jennies hd similr vlues compred to mres throughout the study period [37,42]. The PLT trend showed lower vlues from delivery to the first two months of lcttion compred to lte gesttion, thus the post-prtum period seems to exert significnt effects on circulting pltelet numers. These results re not in greement with previous studies on mres where PLT were found to e constnt over time [32,38-39]. The results on MCV, MCH nd MCHC showed constnt tendency over time, s reported for mres [39], ut with higher vlues, while MCHC ws similr for the two species during the ll study period [37-38,42]. Glucose concentrtion ws constnt throughout the study period in jennies nd vlues were in line to those reported in pregnnt nd lctting mres [42], ut higher compred to dult donkeys [6,15]. This could e due to the development of insulin resistnce, s reported for horses [39,51] nd dogs [52], s well s for women [53]. Blood ure incresed ner prturition (-2 weeks), nd then remined constnt during delivery nd lcttion. This trend might e due to the high-energy demnd t the eginning of lcttion, s oserved in mres [42]. 8

9 Cretinine vlues were constnt throughout the study period nd within norml rnges for dult species [15,17-18,20-21,26], thus jennies do not seem to e influenced y the increse in energy demnd in lte gesttion nd y the quot produced y the foetus, s reported in mres [42]. Triglycerides nd totl cholesterol concentrtions showed n importnt decrese from delivery through the lcttion period, in prticulr two weeks fter prturition. Our dt re in line with previous studies on pony nd drft mres in which triglycerides showed lower vlues during lcttion thn efore prturition [41,54]. However they re in contrst with studies on light reed mres [29], in which triglycerides were constnt nd concentrtions were lwys similr to reference vlues for dult horses. Our results showed more similr metolism of fts to ponies nd drft horse mres, which hve greter ft content nd moiliztion thn light reed horses. Totl iliruin ws constnt throughout the study period. In mres, totl iliruin increses in lte pregnncy due to secondry cholestsis ecuse of the enlrged uterus [42], while in jennies this does not seem to occur. The AST ctivity trend showed n increse ner to prturition nd erly lcttion. This might e due to predominnce of nolic over ctolic processes during pregnncy [55]. Compred to ctivity vlues otined in mres, our results were lwys higher [38,42]. The GGT ctivity trend showed lower vlues t delivery nd during lcttion thn lte gesttion. Our results re in contrst to findings in mres in which the GGT ctivity incresed round delivery nd decresed grdully fter foling [55], which could e relted to physiologicl lod on the liver in the perintl period. GGT ctivity in jennies ws lwys higher thn in mres [42]. CK decresed ner delivery, thn remins constnt fter prturition for the first two months of lcttion. These results re not in greement with Mrell et l. [42], who found n increse in CK ctivity t delivery nd during lcttion compred to pregnncy. It is difficult to explin the decresed ctivity of this enzyme ecuse to the uthors knowledge there re no studies on CK ctivity during pregnncy, lcttion nd prturition in donkeys. ALP ctivity vlues were constnt nd in line with pregnnt nd lctting light reeds mres [39], ut not with drft mres in which ALP ctivity ws found to increse round delivery nd decrese grdully fter foling [41]. Tot C vlues were constnt throughout the study period, while decrese hs een oserved in mres t prturition [42]. P ws higher in lte gesttion nd prturition thn in 9

10 lctting jennies. To the uthors knowledge, no dt hve een reported for P either in jennies or mres, ut our results re in line with findings reported for women [56-57]. N decresed during lcttion nd concentrtions were lwys comprle to donkey reference intervls [21]. The higher N concentrtions during pregnncy might e due to n incresed renl N retention medited y ldosterone relese during pregnncy, s lredy reported in mres [58]. K showed the sme trend s N nd concentrtions re in line with this species [21]. The higher K vlues during pregnncy in jennies my e relted to the resorption y the gut due to the enlrged pregnnt uterus, s suggested for mres [42]. The TP trend showed decresed concentrtion during the post-prtum period nd lcttion thn the gesttionl period, in contrst with mres in which higher TP concentrtions were found in lte pregnncy nd erly lcttion compred to prturition [42]. In conclusion, our results showed significnt chnges in hemtologicl nd iochemicl prmeters in jennies during the lst two months of pregnncy nd the first two months of lcttion. These chnges re only prtilly comprle to mres. Thus, vlues otined in jennies could e useful in clinicl prctice to ssess the helth sttus in jennies nd to check peri-prtum diseses. The results contriute to etter understnding of the iochemicl processes in pregnnt nd lctting jennies, in order to estimte their physiologicl sttus nd to e used for dignostic purposes. 10

11 Acknowledgements The uthors wish to thnk the Regionl Stud Centre of Tuscny, Itly. This work ws supported y MIUR funds. References [1] Crroccio A, Cvtio F, Montlto G. Intolernce to hydrolysed cow s milk proteins in infnts: clinicl chrcteristic nd dietry tretment. Clin Exp Allergy 2000; 30: [2] Murro MA, Gimpietro PG, Glli E. Soy formuls nd non ovine milk. Ann Allergy Asthm Immunol 2002; 89(S): [3] Borioni M, Mrinro P, Celestini S, Del Sole F, Mgro R, Zoppi D, et l. Effect of equestrin therpy nd onotherpy in physicl nd psycho-socil performnces of dults with intellectul disility: preliminry study of evlution tools sed on the ICF clssifiction. Disil Rehil 2012, 34(4): [4] Nyieri GD. Blood chrcteristics of the dult donkey. Zl Vet Med A 1978; 25: [5] Zinkl JC, Me D, Guzmn P, Frver TB, Humle JA. Reference rnges nd the influence of ge nd sex on hemtologicl nd serum iochemicl vlues in donkey (Equus sinus). Am J vet Res 1990; 51: [6] French JM, Ptrick VH. Reference vlues for physiologicl, hemtologicl nd iochemicl prmeters in domestic donkeys (Equus sinus). Equine vet Educ 1995; 7: [7] Orlndi M, Curdi MC, Leott R, Berni P. Metolic profile in the Amit donkey. Annli Fcoltà Medicin Veterinri di Pis 1997; L: ISSN: [8] Folch P, Jordn J, Cuenc R. Reference rnges nd the influence of ge nd sex on hemtologicl vlues of the endngered Ctlonin donkey. Vet J 1997; 154: [9] Jordn J, Folch P, Cuenc R. Clinicl iochemicl prmeters of the endngered Ctlonin donkey reed: norml vlues nd the influence of sex, ge, nd mngement prctices effect. Res Vet Sci 1998; 64: [10] Mushi EZ, Bint MG, Ndeele RT. Hemtologicl studies on pprently helthy donkeys in Oodi, Kgtleng district Botswn. Proceedings of n ATNESA workshop, South Afric, 1999;

12 [11] Mori E, Fernndes WR, Mirndol RMS, Kuo G, Ferreir RR, Oliveir JV, Gcek F. Reference vlues on serum iochemicl prmeters of Brzilin donkey (Equus sinus) reed. J Equine Vet Sci 2003; 23(8): [12] Crcngiu V, Vcc GM, Cueddu GM, Bini PP. Rilievi emtici in sini inchi dell Asinr sottoposti trttmento per il controllo delle ectoprssitosi. J Mt Ecol 2003; 7(S): [13] Mori E, Mirndol RMS, Ferreir RR, Oliveir JV, Gcek F, Fernndes WR. Reference vlues on hemtologic prmeters of the Brzilin donkey (Equus sinus) reed. J Equine Vet Sci 2004; 24(7): [14] Al-Busdh KA, Homeid AM. Some physicl vriles, iochemicl nd hemtologicl prmeters in Hsswi ss. Scientific Journl King Fisl University (Bsic nd Applied Sciences) 2005; 6(1): [15] Cldin M, Furlnello T, Solno-Gllego L, De Lorenzi D, Crli E, Tsc S, et l. Reference rnges for hemtology, iochemicl profile nd electrophoresis in single herd of Rgusn donkeys from Sicily (Itly). Comp Clin Pth 2005; 14: [16] Lopez I, Estep JC, Mendoz FJ, Rodriguez M, Aguiler-Tejero E. Serum concentrtions of clcium, phosphorus, mgnesium nd clciotropic hormones in donkeys. J Am Vet Res 2006; 67(8): [17] Pitel PH, Moulin M. Approche des vleurs hemtologiques et iochimiques chez deux rces sinines. Prtique Veterinire Equine 2006; 38(149): [18] De Aluj AS, Mondrgon RL, Cstillo DA, Ocho P. Hemtologicl nd iochemicl reference vlues in the donkey (Equus sinus) in Mexico. In: Mtthews NS, Tylor TS, editors. Veterinry Cre of Donkeys, Interntionl Veterinry Informtion Service, Ithc, New York, ( Lst updte: 21/06/2006. [19] Ahemed AE, Adel-Hmid HY, Adel-Rhim AA, Ismil MN. The influence of ge nd sex on some lood prmeters in helthy donkey in south Vlley Egypt. Bs Vet Med J, 2007; [20] Couroucè-Mllnc A, Fortier G, Moulin M, Vlette JP, Petit L, Dumontier S, et l. Reference vlues on hemtologic nd iochemicl prmeters in french donkeys. Proceedings 10 th WEVA 2008; [21] Lemm A, Moges M. Clinicl, hemtologicl nd serum iochemicl reference vlues of working donkeys (Equus sinus) owned y trnsport opertors in Addis A, Ethiopi. Livest Res Rurl Dev 2009; 21(8):

13 [22] Etn KM, Jenere TS, Bojis E, Negussie H. Determintion of Reference Hemtologicl nd Serum-Biochemicl Vlues for Working Donkeys of Ethiopi. Vet Res 2011; 4(3): [23] Alerghin D, Fzio F, Arfuso F, Scinò S, Zumo A, Piccione G. Reference Intervls of Serum Protein Concentrtions from Cliniclly Helthy Femle Rgusn Donkeys (Equus sinus) Determined y Cellulose Acette Electrophoresis. J Equine Vet Sci 2013; 33(6): [24] Zkri FO, Ayo JO, Rekwot PI, Kru MU. Effects of ge nd seson on hemtologicl prmeters of donkeys during the riny nd cold dry sesons. Int J Biometereol 2015; /s [25] Moris Girrdi A, Mrques LC, Pereir de Toledo CZ, Bros JC, Mldondo Jr W, Lemos Ngi Jorge R, Aprecid d Silv Nogueir C. Biochemicl profile of the Pêg donkey (Equus sinus ) reed: influence of ge nd sex. Comp Clin Pthol 2014; 23(4): [26] Lus F, Sptern A, Fillce V, Pggi E, Serri E, Vullo C, Cerquetell M, Tesei B. Reference vlues for hemtologicl nd iochemicl prmeters of mixed reed donkeys (Equus sinus).wulfeni J 2015; 22(1): [27] Sgorini M, Bonelli F, Rot A, Brgli P, Mrchetti V, Corzz M. Hemtology nd Clinicl Chemistry in Amit Donkey Fols from Birth to 2 Months of Age. J Equine Vet Sci 2013; 33(1): [28] Veronesi MC, Glori A, Pnzni S, Sfirro MP, Crluccio A. Blood nlysis in neworn donkeys: hemtology, iochemistry nd lood gses nlysis. Theriogenol 2014; 82(2): [29] Hrvey JW, Pte M, Kivipelto J, Asquisth R. Clinicl iochemistry of pregnnt nd nursing mres. Vet Clin Pthol 2005; 3(3): [30] Lrsson A, Plm M, Hnsson LO, Axelsson O. Reference vlues for clinicl chemistry test during norml pregnncy. Int J Ostet Gynecol 2008; 115: [31] Butte NF. Crohydrte nd lipid metolism in pregnncy: norml compred with gesttionl dietes mellitus. Am J Clin Nutr 71: [32] Hfez SA, Freemn LE, Cceci T, Smith BJ. Study of the vsculture of the cprine reproductive orgns using the tissue clering technique, with specil reference to the ngiorchitecture of the utero ovrin vessels nd the dpttion of the ovrin nd/or vginl rteries to multiple pregnncies. Ant Rec 2007; 290:

14 [33] Stué K, Montesinos P. Plsm iochemistry profile in pregnnt Spnish purered roodmres. Comp Clin Pthol 2013; 22: [34] Hnsen MF, Todd, AC, McGee WR. Blood picture of lctting nd non lctting Throughred mres. Vet Med 1950; 45: [35] Meuten DJ, Koci G, Therelfll WR, Ngode LA. Serum lkline phosphtse in pregnnt mres. Vet Clin Pth, 1980; 9: [36] Gill J, Kompnowsk-Jeziersk E. Sesonl chnges in the red lood cell indices in Arin rood mres nd their fols. Comp iochem physiol 1986; 83(4): [37] Hrvey JW, Asquith RL, Pte MG, Kivipelto J, Chen CL, Ott EA. Hemtologicl findings in pregnnt, postprturient nd nursing mre. Comp Hemtol Int 1994; 4: [38] Tylor-Mcllister C, Mcllister CG, Wlker D, Alseth D. Hemtology nd serum iochemistry evlution in norml postprtum mres. Equine vet J 1997; 29 (3): [39] Hoffmn RM, Kronfeld DS, Cooper WL, Hrris P. Glucose clernce in grzing pregnnt mres is ffected y diet, pregnncy nd lcttion. J Anim Sci 2003; 81: [40] Berlin D, Aroch I. Concentrtions of ionized nd totl mgnesium nd clcium in helthy horses: Effects of ge, pregnncy, lcttion, ph nd smple type. Vet J 2009; 181: [41] Aoki T, Ishii M. Hemtologicl nd iochemicl profiles in periprtum mres nd neontl fols (Hevy drft horses). J Equine Vet Sci 2012; 32: [42] Mriell J, Pirrone A, Gentilini F, Cstgnetti C. Hemtologicl nd iochemicl profiles in Stndrdred mres during periprtum. Theriogenol 2014; 81(4): [43] Crisci A, Rot A, Pnzni D, Sgorini M, Ousey JC, Cmillo F. Clinicl, ultrsonogrphic, nd endocrinologicl studies on donkey pregnncy. Theriogenol 2014; 81: [44] Aville t: Accessed April 10,

15 [45] Gurgoze S, Icen H. The influence of ge on clinicl iochemicl prmeters in pure-red Arin mres. J Equine Vet Sci 2010; 30(10): [46] Bilit JL, Dty E, Jodie W. Repeted hemtocrit mesurements in low-risk pregnnt women. J Reprod Med 2007; 52(7): [47] Allrd RL, Crlos AD, Fltin EC. Cnine hemtologicl chnges during gesttion nd lcttion. Compn Anim PRct 1989; 19: 3-6. [48] Bermn E. Hemogrms of the ct during pregnncy nd lcttion nd fter lcttion. Am J Vet Res 1974; 35: [49] Wells MY, Decoecq CP, Decouvelere DM, Justice C, Guittin P. Chnges in clinicl pthology prmeters during gesttion in the New Zelnd white rit. Toxicol Pthol 1999; 27: [50] Blnchrd TL, Orsini JA, Grci MC, Elmore RG, Youngquist RS, Bierschwl CJ. Influence of dystoci on white lood cell nd lood neutrophil counts in mres. Theriogenol 1986; 25: [51] Evns JW. Effect of fsting, gesttion, lcttion nd exercise on glucose turnover in horses. J ANim Sci 1071; 33: [52] Johnson CA. Glucose homeostsis during cnine pregnncy: insulin resistnce, ketosis, nd hypoglycemi. Theriogenol 2008; 70(9): [53] Dhlgren J. Pregnncy nd insulin resistnce. Met Syndr Relt Disord 2006; 4(2): [54] Wtson TD, Burns L, Pckrd CJ, Sheperd J. Effect of pregnncy nd lcttion on plsm lipid nd lipoprotein concentrtions, lipoprotein composition nd post-heprin lipse ctivities in Shetlnd pony mres. J Reprod Fertil 1993; 97: [55] Dvies Morel MC. Equine reproductive physiology, reeding nd stud mngement. Ipswich, Englnd: Frming Press Books; [56] Sultn MS, Begum R, Akhter KS, Lovely NS, Akhter S, Islm MT. Serum clcium nd phosphte level in norml pregnnt women. Bngldesh Journl of Medicl Science; 2012; 11(3): [57] Mnsmnn, HC Jr. Phosphorus, Clcium nd Mgnesium Interctions. 15

16 416 RBC M/µL WBC K/µL Hg gr/l Hct % MCV fl MCH pg MCH C gr/dl PLT K/µL T-8w T-6w T-4w T-2w T0 T2w T4w T6w T8w 5.7± ± ± ± ± ± ± ± ±0.6 P= ± ± ± ± ± ± ± ± ±1.9 P= ± ± ± ± ± ± ± ± ±1.3 NS 33.2± ± ± ± ± ± ± ± ±3.7 P= ± ± ± ± ± ± ± ± ±1.6 NS 20.1± ± ± ± ± ± ± ± ±0.8 NS 35.7± ± ± ± ± ± ± ± ±1.6 NS 252.0± ± ± ± ± ± ± ± ±50. P= n= Tle 1. Hemtologicl prmeters expressed s men± stndrd devition in 9 jennies from 2 months efore delivery up to 2 months fter delivery, for totl of 18 pregnncies nd 16 fols orn. Legend - T-8w: 8 weeks efore presumptive delivery; T-6w: 6 weeks efore presumptive delivery; T-4w: 2 weeks efore presumptive delivery; T-2w: 2 weeks efore presumptive delivery; T0: delivery; T2w: 2 weeks fter delivery; T4w: 4 weeks; T4w: 4 weeks fter delivery; T6w: 6 weeks fter delivery; T8w: 8 weeks fter delivery. Within row, different superscripts denote significnt difference ( : P<0.05)

17 426 Glucose mg/dl Cretinine mg/dl Ure mg/dl Triglycerid e mg/dl Totl cholesterol mg/dl Totl iliruin mg/dl AST U/L T-8w T-6w T-4w T-2w T0 T2w T4w T6w T8w 91.0± ± ± ± ± ± ± ± ±8.9 NS 1.2± ± ± ± ± ± ± ± ±0.2 NS 25.3± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ± ±15.0 c 52.3± ±18.7 c 50.7± ±9.5 P= ±25.7 P= ±7.0 c 64.3±5.7 P= ± ± ± ± ± ± ± ± ±0.04 NS 82.9± ± ± ± ± ± ± ± ±36. P= GGT U/L 38.6± ± ± ± ± ± ± ± ±10.8 P=0.001 CK U/L 60.6± ± ± ± ± ± ± ± ±14.7 P=0.003 ALP U/L 168.6± ± ± ± ± ± ± ± ±32. 4 NS Totl C mg/dl P mg/dl N + mmol/l 11.4± ± ± ± ± ± ± ± ±1.4 NS 3.7± ± ± ± ± ± ± ± ±0.2 P= ± ± ± ± ± ± ± ± ±4.1 P= K + 4.3± ± ± ± ± ± ±0.5 c 4.3±0.4 c 3.7±0.5c P=0.03 mmol/l PT gr/l 8.2± ± ± ± ± ±0.5 c 7.1±0.4 c 6.9±0.9 c 7.3±0.7 c P= Alumin 2.8± ± ± ± ± ± ± ± ±0.7 NS gr/l n= Tle 2. Clinicl chemistry prmeters expressed s men±stndrd devition in 9 jennies from months efore delivery up to 2 months fter delivery, for totl of 18 pregnncies nd 16 fols orn. Legend - T-8w: 8 weeks efore presumptive delivery; T-6w: 6 weeks efore presumptive delivery; T-4w: 2 weeks efore presumptive delivery; T-2w: 2 weeks efore presumptive delivery; T0: delivery; T2w: 2 weeks fter delivery; T4w: 4 weeks; T4w: 4 weeks fter delivery; T6w: 6 weeks fter delivery; T8w: 8 weeks fter delivery. Within row, different superscripts denote significnt difference ( c c: P<0.05)