Blood clinicopathological differences between type I and II ketosis in dairy cows

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1 ndin J. nim. Res., 50 (5) 2016 : Print SSN: / Online SSN: GRULTURL RESERH OMMUNTON ENTRE lood clinicopthologicl differences etween type nd ketosis in diry cows XU hung*, Shen Ti-yu, YO Yun, Yu Hong-jing, X heng nd Zhng Hong-you 1 ollege of niml Science nd Veterinry Medicine, Heilongjing yi griculturl University, Dqing, Heilongjing , hin. Received: ccepted: DO: /ijr.7495 STRT The purpose ws to determine the difference of lood clinicopthologicl chnges etween type nd type ketosis in diry cow. Fifty-eight cows, from diry cttle frm in Heilongjing of hin, were included. n ELS test ws used to evlute the lood indictors. The plsm concentrtions of et-hydroxyutyric cid (H) nd insulin sensitivity decresed, nd the plsm concentrtion of glucose (Glc), non-esterified ftty cid (NEF) nd iliruin content incresed in type ketosis group compred with the type ketosis group. These results showed tht there ws difference in etiology etween type ketosis nd type ketosis. Type ketosis ws not only ssocited with energy metolism nd insulin resistnce, ut lso with oxidtive stress nd liver function. t lid the foundtion for further investigte the mechnism nd prevention of type ketosis in the future. Key words: Diry cow, Energy metolism, nsulin resistnce, Liver function, Oxidtive stress, Type ketosis. NTRODUTON During the trnsition diry cow undergo period of negtive energy lnce for the demnds of milk synthesis, which cnnot e met y feed intke (Wng et l., 2010). Ketosis, one of the most prevlent metolic diseses in diry cows during trnsition period, ws condition tht the concentrtion of ketone in lood ws exceeding thn 3,000 mol/l (Oetzel, 2007) during the trnsition (Shire et l., 2013; Zhng et l., 2013). ows with ketosis were chrcteristic with dry mtter intke (DM) nd milk production decresed, nd reduced reproductive performnce. n ddition, it ws known tht ketosis ws t n incresed risk of dditionl postprtum diseses such s displced omsum (D), ftty liver nd metritis, which my increse their risk of removl from the her during erly lcttion (de Roos et l., 2007; tle et l., 2015). From n etiologicl viewpoint, there were two quite different types of metolic disorders in which ketosis cn occur, type ketosis nd type ketosis, respectively (Holtenius nd Holtenius 1996). Type, generlly occurs 3-6 weeks fter clving in cows whose milk secretion is so extensive tht the demnd for glucose exceeds the cpcity for glucose production, ws chrcteristic with decrese in ppetite nd milk production, nd weight loss. n ddition, there ws decresed on glucose, ut the concentrtions of et-hydroxyutyric cid (H) nd non-esterified ftty cid (NEF) were incresed (erge nd Vertenten 2014).However, type, generlly occurs erlier in lcttion, leds to the concentrtions of H, glucose nd NEF were incresed (Oikw nd Oetzel 2006). *orresponding uthor s e-mil: xuchung7175@163.com t ws reported tht the prevlence of ketosis ws 5.0% in meric; 14.7% in ndi; 43.1% in Jpn, nd 15-30% in hin. nd there ws $ per diry cow losing on the therpy (Ke-he 2008). Therefore, we mde n investigtion on the diry frm, nd found tht there were higher incidence on type ketosis thn type ketosis. nd the mechnism of type ketosis ws uncler, thus, n experiment ws conduct to provide review of differences in diry cows with type nd ketosis on the energy metolic, liver function, stress oxidtion nd insulin resistnce. MTERLS ND METHODS Ethics sttement: The study ws pproved y the frm owner nd ll experimentl nimls were conducted ccording to the nterntionl Guiding Principles for iomedicl Reserch. The protocol ws pproved y the ommittee on the Ethics of niml Experiments of the Heilongjing yi griculturl University. nimls nd Groups: Fifty-eight Holstein cows were selected from n intensive 2000 diry cttle frm in Heilongjing, hin. ll cows were 3 to 5 yer old. ll cows hd ody condition score (S) etween 2.75 nd S ws recorded y the sme person immeditely fter prturition. S ws expressed on 5-point scle with 0.25 increments s descried y Edmonson et l. (1989). ows were divided in 3 clsses sed on their S: (1) underconditioned, <3; (2) norml condition, 3 to 3.5; nd (3) overconditioned, >3.5(Megnck, et l. 2015). ll the cows were fed totl mixed rtion (TMR) t 4:00 m, 10:00pm nd

2 754 NDN JOURNL OF NML RESERH 16:00 pm dily during trnsition period, which consisted of concentrted feed, silge corn, rewer s grin, cooked soyens, hinese hy, melon pulp, surfce melon shell, nd ft. The nutritionl nlysis ws 55.6% dry mtter (DM), 17% crude protein, 1.75mcl/DM net energy for lcttion (NEL), 5.60% ft, 39.10% neutrl detergent fier (NDF), 20.60% cid detergent fier (DF), 180g, nd 116g P. ccording to the concentrtions of Glc, NEF, H nd the clinicl symptoms the cows were clssified into three groups y using the clssify method of Holtenius, ontrol group, Type ketosis group ( group) nd type ketosis group ( group), respectively. The clssifiction stndrd is tht if the concentrtion of Glc more thn 3.75, H concentrtion ws less thn 0.6 mmol/ L, NEF concentrtion ws less thn 0.5, descriing it s control group; if the concentrtion of Glc less thn 2.5, H concentrtion exceeds 1.2, NEF concentrtion greter thn 0.5, descriing it s type ketosis, otherwise, the concentrtion more thn 2.8, nd other remining indictors re equl, descriing it ws type ketosis. There were 12 cows which were not occurred ketosis during 0-28 dy, divided them into group. On the 7 dy fter culling, there re 6 cows with type ketosis, nd 8 cows with type ketosis. nd there re 8 cows with type ketosis nd 12 cows with type ketosis fter 14 dys. There were 10 cows with type or type ketosis on the dy of 21 dy. There re 5 cows with type ketosis nd 7 cows with type ketosis on the 28 dy. Smple collection: lood smples were collected from the ven cudlis medin using sodium heprin t the time of dy of prturition, nd gin t 0d, 7d, 14d, 21d nd 28d fter prturition nd then immeditely centrifuged t 1,400 g for 10 min t room temperture. The superntnts were liquoted into Eppendorf tues (1 ml plsm/tue) nd stored t -80 until nlysis. Tle 1: The prmeters of two type ketosis nd helthy cows Prmeters Groups Time of postprtum (dy) Prmeters nd method : linicl lood iochemicl dt included glucose (Glc, Oxidse method, purchsed from iosino io-technology nd Science N), ethydroxyutyric cid (H, Enzyme rte method, purchsed from eijing jiuqing iotechnology ompny), nd non-esterified ftty cids (NEFs, colorimetric method, purchsed from Sekisui Medicl Technology(hin)) for ssessing energy lnce sttus, nd sprtte minotrnsferse (ST), lnine minotrnserse (LT), totl iliruin (TL, were devised y kit method, purchsed from iosino io-technology nd Science N)for mesuring liver function, nd insulin (NS, Elis, purchsed from Nnjing jincheng Technology ompny) nd diponectin (DP, Elis, RD ompny in meric) for evlution the insulin resistnce sttus, nd then mlondildehyde (MD) nd nitric oxide (NO) (Nitric cid reduction method)nd SOD (Hydroxylmine method) they were purchsed from Nnjing jincheng technology ompny for determining oxidtive stress sttus. Sttistics nlysis: The dt ws used SPSS17.0 softwre for sttisticl nlysis using the single fctor vrince; the results were expressed s mens ± stndrd devition. The revised quntittive insulin sensitivity check index, (RQUK), following formul: RQUK=1/ [log (glucose in mg/dl) +log (insulin in mu/ml) +log (NEF in )]. RESULTS ND DSUSSON The cows were divided into three groups ccording the dtes from Tle 1, the Glc concentrtion of type ws significntly higher thn type ketosis. nd there were lso differences etween H nd NEF, nd they were ll significnt higher thn group. n ddition, H nd Glc got the pek t the 14d; NEF got the pek t the 7d in the cows with type ketosis. nd H, Glc nd NEF got the pek t the 7d in the cows with type ketosis. The figure 1 showed tht the concentrtions of lnine minotrnsferse (LT) nd sprtte mino trnsferse (ST) of type ketosis were higher thn type ketosis on the 7-28 dys, nd they were ll significnt higher thn group. The totl iliruin (TL) concentrtion of type ketosis ws higher thn type ketosis on the 7 dy t postprtum, nd there ws no significntly difference t the others dy. H 0.511± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.17 Glc 4.706± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.32 NEF 0.57± ± ± ± ± ± ± ± ± ± ± ± ± ± ±0.12 Note: the groups were divided compred with control diry cows. Different cpitl letters in sme row represent very significnt difference (P <0.01), different lowercse letters represent significnt difference (0.01<P <0.05), nd the sme letter or without letters men no significnt difference (P > 0.05).

3 The NS concentrtion of type ws significntly higher thn group, nd there ws no significntly difference etween the type nd ketosis t the 0-28dy. The DP concentrtion ws decresed compred with type ketosis Volume 50 ssue 5 (2016) 755 except the 21dy. The concentrtion of RQUK ws significntly lower thn the type ketosis on the 0-8 dy (Fig 2). From the figure 3, the trend of superoxide dismutse (SOD) concentrtion were first incresed nd then decresed, LT TME(d) LT c ST S TME(d) c TL TL TME(d) Fig 1: The prmeters of liver function nd the ketosis in the diry cows Note: Liver function nd the cows with ketosis. The trnsverse coordinte express the time of postprtum. Longitudinl coordinte express the concentrtions of ST (sprtte minotrnsferse), LT (lnine minotrnsferse) nd TL (Totl iliruin). Different cpitl letters in sme row represent very significnt difference (P <0.01), different lowercse letters represent significnt difference (0.01<P <0.05), nd the sme letter or without letters men no significnt difference (P > 0.05). mmol/l NS TME(d) RQUK RQUK TME(d) DP DP 220 TME(d) Fig 2: The prmeters of insulin resistnce nd the ketosis in the diry cows Note: nsulin resistnce nd the cows with ketosis. The trnsverse coordinte express the time of postrtum. Longitudinl coordinte express the concentrtions of DP (diponectin), NS (insulin) nd RQUK (Revised quntittive insulin sensitivity). Different cpitl letters in sme row represent very significnt difference (P <0.01), different lowercse letters represent significnt difference (0.01<P <0.05), nd the sme letter or without letters men no significnt difference (P > 0.05).

4 756 NDN JOURNL OF NML RESERH nd got the pek on the 14 dy oth two types ketosis. There ws significnt deference in mlondildehyde (MD) concentrtion t the 0-7 dys, nd the content of type ketosis ws higher thn the type ketosis. The other dys there ws no significntly different. There ws no significntly difference on the nitric oxide (NO) etween type nd ketosis. The trend of concentrtion ws firstly decresed nd then incresed, nd oth of them got the pek t the 21 dy. Energy metolic nd type ketosis: During the perintl period diry cows undergo pregnncy, culling nd lcttion. s result of nutrient demnd for milk productions incresing fster thn DM, cows experienced period of negtive energy lnce (NE). To dpt to this negtive energy lnce, metolic disorder occurred in this time (ouwstr et l., 2008; González et l., 2011; Sntschi et l., 2011; Mcrt et l., 2012). H nd NEF re the products of ft metolism. H content reflects the oxidtion of ft metolic, nd NEF content reflects the moiliztion of ft. When the contents of H nd NEF incresed in the lood, it indicts tht there re lot of moiliztion of ft (Li et l., 2012). The diry cow induced ketosis when the concentrtion of NEF product more thn the demnds of ody. The concentrtions of H nd NEF re higher thn the norml in the figure 1, it reflects tht the ody is in negtive energy lnce. Glc minly offer energy, once the Glc concentrtion is lower thn the demnd for the milk product, NEF will e moiliztion (oe et l., 2004). From figure 1 there re significnt difference etween type nd type ketosis. The Glc concentrtion of type ketosis is higher thn the type ketosis, nd higher or equl with norml diry cow. Liver function nd type ketosis: The liver minotrnsferses-lt nd ST re oth found in the liver, serum nd vrious orgn tissues. LT nd ST re found predominntly in the liver nd its serum levels ecome elevted whenever disese process ffects liver cells (Vozrov et l., 2002; Kunutsor et l., 2013). From figure 2 there re higher LT nd ST concentrtions in type ketosis thn type ketosis, nd they re ll higher thn norml level, it indicts tht there re liver dmge when hppened ketosis whether type or type, nd type ketosis hs igger dmge thn type ketosis. Totl iliruin (TL) is produced s n unconjugted from y hemogloin fter the end of life of red lood cells from the reticuloendothil system such s in spleen. The iliruin is incresed fter dmge of liver due to infection (Ginnini et l., 2005). The concentrtion of TL of type ketosis is more thn type ketosis on the 0-14 dys of prtum. nd they re ll incresed compred with the norml level of diry cow. t suggests tht there is dmge in the liver. nsulin resistnce nd type ketosis: nsulin is secreted y the pncres, it minly plys role of liver nd peripherl tissue. When the liver dmged, the sensitivity of insulin chnged (ossert et l., 2008). diponectin (DP), n dipose-specific secretry protein, exhiits ntidietic nd ntitherogenic properties (Nishizw et l., 2002). nd DP SOD TME(d) MD TME(d) NO NO TME(d) Fig 3: The sttus of stress oxidtion nd ketosis in diry cow Note: Stress oxidtion nd the cows with ketosis. The trnsverse coordinte express the time of postprtum. Longitudinl coordinte express the concentrtions of SOD (Superoxide dismutse), MD (Mlondildehyde) nd NO (Nitric oxide). Different cpitl letters in sme row represent very significnt difference (P <0.01), different lowercse letters represent significnt difference (0.01<P <0.05), nd the sme letter or without letters men no significnt difference (P > 0.05).

5 plys n importnt role in the ccumulted dipose tissue, it is thought to e prtilly responsile for the development of insulin resistnce (R) in oesity. R is defined s stte in which norml concentrtion of insulin produce less thn norml iologicl response (Zchut et l., 2013). R my e the min reson wht cuse the high glucose in diry cow. Sttes of R my consist of decresed insulin sensitivity, decresed insulin responsiveness on comintion of oth decresed insulin sensitivity nd responsiveness (Oetzel 2006). From figure 2, there ws significntly difference etween type ketosis nd group, nd the NS concentrtion of type ws higher thn type ketosis. n ddition, the DP concentrtion of type ketosis is lower thn type ketosis. t ll suggested tht the cows with type ketosis exists R. t grees with the other studies. Oxidtion stress nd type ketosis: Diry cow with ketosis experienced oxidtive stress which cn e mesured in iologicl fluids nd tissues. MD is one of the finl products of polyunsturted ftty cid peroxidtion, which is cused y incresing level of rective oxygen species (ROS) nd is commonly known s mrker of oxidtive dmge (Wng et l., 2013). nd SOD is considered the first intrcellulr defense ginst rective oxygen molecules. t cuses dismuttion of the superoxide nion rdicl to H2O2, which is further degrded y T nd peroxidse ctions (Schogor et l., 2013). nd the NO is n only free rdicl with the gs form in the ody, nd it mintins mucosl vsodilttion nd vsculr permeility. The excess cn e directly Volume 50 ssue 5 (2016) 757 cytotoxic nd lso rect with superoxide nions resulting in peroxyntrites tht cn cuse tissue dmge (hoe et l., 2003). From figure 3, the concentrtion of MD in type ketosis is higher thn type on the 0-28d, it indicts tht the cows with type ketosis is in the oxidtion. NO contents of type ketosis is higher thn type ketosis on the 0-28 dy, it showed tht the oxidtion of type ketosis enhnced thn type ketosis. nd then the concentrtions of the MD nd NO incresed, it indicts the cpcity of oxidtion enhnced. Overll, The Oxidtion nd ntioxidtion system imlnced in diry cow with the type ketosis. ONLUSON To sum up, study on the energy metolic, liver function, insulin resistnt nd stress oxidtion showed tht type ketosis with higher glucose nd NEF, lower H, liver dmged, emergent insulin resistnt nd within oxidtion sttement compred with type ketosis. These lood clinicopthologicl chnges of type Ketosis ply n importnt role in the further study of pthogenesis of type ketosis, nd it lso hve significnce mens to mke new mesure to prevent the ketosis. KNOWLEDGEMENTS The uthors thnk finncil support from New entury Excellent Tlents in University (1252-NET-003), the hinese Ntionl Nture Science Foundtion ( ÿ ), the Fok Ying-Tong Eduction Foundtion, hin (141021) nd hinese Ntionl Nture Science Foundtion (2013D2101ÿ2012D1205). REFRENES erge,. nd Vertenten, G. (2014). field study to determine the prevlence, diry herd mngement systems, nd fresh cow clinicl conditions ssocited with ketosis in western Europen diry herds. J Diry Sci 97: oe, G., Young JW. nd eitz, D. (2004). nvited review: pthology, etiology, prevention, nd tretment of ftty liver in diry cows. Journl of Veterinry Science 87: ossert, P., Leroy JL., De Vliegher S nd Opsomer G (2008). nterreltions etween glucose-induced insulin response, metolic indictors, nd time of first ovultion in high-yielding diry cows. J Diry Sci 91: ouwstr, RJ., Goselink RM, Doelr P, Nielen M, Newold JR. nd vn Werven T (2008). The reltionship etween oxidtive dmge nd vitmin E concentrtion in lood, milk, nd liver tissue from vitmin E supplemented nd nonsupplemented periprturient heifers. J Diry Sci 91: hoe, W., Kim S, Hwng TS, nd Lee SS (2003). Expression of inducile nitric oxide synthse in thyroid neoplsms: immunohistochemicl nd moleculr nlysis. Pthology nterntionl 53: De Roos, P., vn den ijgrt HJ., Horlyk J, nd de Jong G (2007). Screening for suclinicl ketosis in diry cttle y Fourier trnsform infrred spectrometry. J Diry Sci 90: Ginnini, EG., Test R, nd Svrino V (2005). Liver enzyme ltertion: guide for clinicins. MJ 172: González, FD., Muiño R, Pereir V, mpos R, nd enedito JL. (2011). Reltionship mong lood indictors of lipomoiliztion nd heptic function during erly lcttion in high-yielding diry cows. Journl of Veterinry Science 12: 251. Holtenius, P. nd Holtenius K (1996). New spects of ketone odies in energy metolism of diry cows: review. Journl of Veterinry Medicine Series 43: tle, J., Huzzey JM, Wery DM. nd von Keyserlingk M. (2015). linicl ketosis nd stnding ehvior in trnsition cows. J Diry Sci 98: Ke-he, H. (2008). The progrss of ketosis nd ftty liver syndrome in diry cows. hin Diry 6: Kunutsor, SK., pekey T. nd Wlley J (2013). Liver minotrnsferses nd risk of incident type 2 dietes: systemtic review nd met-nlysis. m J Epidemiol 178:

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