Journal of Equine Veterinary Science

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Journal of Equine Veterinary Siene 31 (2011) 645-649 Journal of Equine Veterinary Siene journal homepage: www.j-evs.

Mello MV a, Bruno G. Souza MV, MS a, Fernano Q.

1 Journal of Equine Veterinary Siene 31 (2011) Journal of Equine Veterinary Siene journal homepage: Original Researh Comparison of Insulin Sensitivity of Horses Aapte to Different Exerise Intensities Silvia P. Turner MV, MS a,b, Tanja Maria Hess MV, PhD a,, Kibby Treiber MS, PhD, Eria B. Mello MV a, Bruno G. Souza MV, MS a, Fernano Q. Almeia MV, MS, DS a a Instituto e Zootenia e Instituto e Veterinaria a Universiae Feeral Rural o Rio e Janeiro, RJ, BRAZIL b Universiae Estaio e Sá, Rio e Janeiro, RJ, BRAZIL Equine Sienes, Colorao State University, Fort Collins, CO Nutrition Department, Forth Worth Zoo, TX artile info abstrat Artile history: Reeive 03 January 2011 Reeive in revise form 15 February 2011 Aepte 02 May 2011 Available online 01 July 2011 Keywors: Exerise intensity Insulin Insulin sensitivity Horse Diets high in onentrates an soluble arbohyrates are assoiate with reue insulin sensitivity in horses. Exerise training oul protet against iet-inue insulin resistane. The objetive of this stuy was to etermine the intensity of exerise training require to affet insulin sensitivity in stable horses fe a iet high in onentrates but moerate in soluble arbohyrates. In all, 31 stable horses unerwent three ifferent exerise regimens: turnout, light exerise, an moerate exerise, while being fe a iet ontaining 60% onentrate. Bloo was sample monthly an analyze for insulin. Insulin sensitivity was assesse using basal insulin onentrations an alulate insulin sensitivity (reiproal of the square root of insulin) an ompare aross months by analysis of variane with repeate measures. Insulin sensitivity (reiproal of the square root of insulin) was higher uring perios of moerate an light physial ativity as ompare with turnout. These results iniate that turnout alone may not be aequate to improve insulin sensitivity in horses fe high amounts of onentrate. Ó 2011 Elsevier In. Open aess uner the Elsevier OA liense. 1. Introution Exerise has been shown to improve insulin sensitivity in horses [1,2]; however, it is unlear what exerise intensity is require for this improvement. In young Stanarbre horses, light exerise training (4 weeks) was shown to be as effetive in improving insulin sensitivity as long-term training (18 weeks) with moerate to high intensity [3]. A better unerstaning of the effet of ifferent exerise regimens on insulin sensitivity is neee to evelop effetive management strategies an exerise reommenations by whih owners may reue insulin resistanerelate health risks in their horses. Horses fe iets rih in soluble arbohyrates have an inrease risk of eveloping insulin resistane, obesity Corresponing author at: Tanja Maria Hess, MV, PhD, Equine Siene, Colorao State University, 701 S Overlan Trail, Fort Collins, CO aress: tanja.hess@olostate.eu (T.M. Hess). [4-6], an oli [7]. Diets with moerate amounts of soluble arbohyrates have been shown to avoi ereases in insulin sensitivity [8]. Basal proxies are a noninvasive an ost-effetive tool to evaluate insulin sensitivity in large populations an have been use to etermine insulin sensitivity in horses an other speies [8-10]. The basal proxy reiproal of the square root of insulin (RISQI) reflets the egree of hroni ompensatory hyperinsulinemia that has been negatively orrelate to insulin sensitivity, as etermine by the more rigorous minimal moel [10]. The objetive of the urrent stuy was to ompare ifferent exerise intensities an their effets on insulin sensitivity an basal insulin of horses fe a high ereal-base iet. 2. Materials an Methos The stuy was performe at the Seon Cavalry an Guar Regiment of the Brazilian Army, loate in the ity of Rio e Janeiro, uring the year of A total of Ó 2011 Elsevier In. Open aess uner the Elsevier OA liense. oi: /j.jevs

2 646 S.P. Turner et al. / Journal of Equine Veterinary Siene 31 (2011) mixe bree army horses were weighe on an analyti sale (Coimma, São Paulo, Brazil), sore for boy onition sore (BCS) [11] by two inepenent sorers, an sample for bloo by venipunture between 2:00 an 3:30 AM after 8 hours of fasting. Sampling was performe monthly uring 9 onseutive months (February to Otober of 2007). The stuy was approve by the Animal Care an Use Commission of the Feeral Rural University of Rio e Janeiro Diet an Management Horses were maintaine in iniviual box stalls measuring m 2, unless they were turne out or exerise. All horses were fe iniviually in stalls: 6 kg ommerial onentrate (Royal horse, Brazil) ivie into three aily feeings plus 4 kg of Bermua grass hay ivie in two feeings (Table 1). The afternoon hay feeing was one on the fiel uring turnout months. The feeing regimen was onsistent with the Brazilian Army guieline book [12]. Roughage feeing was restrite beause of low forage availability an quality in this region. High amounts of onentrate were fe to meet nutrient requirements. Diet was offere in amounts inepenent of the horses boy weight or work. Hay an onentrate were sample every 2 months an analyze for nutritional ontent by proximate analysis (ry matter, fat, ash, rue protein, ai etergent fiber, neutral etergent fiber). Hyrolyzable arbohyrates (CHO-H) were analyze by extration with hot water an amylolyti enzymes [13] an represente isaharies, some oligosaharies, an starh [14]. Nonfibrous arbohyrates were estimate by subtrating the sum of rue protein, fat, ash, an neutral etergent fiber from 100 [15]. Rapily fermentable arbohyrates (CHO-F R ) were alulate by subtrating CHO-H from nonfibrous arbohyrates; however, in the CHO-F R fration, organi ais may be inlue [15] Physial Ativity Exerise intensity an turnout time are esribe in Table 2. Exerise sheule was base on military instrution an guar ativities. Before sampling starte, horses were turne out for 8 hours a ay in irt or low grass areas (November of the previous year to January of 2007; Table 2). Horses walke about 1 km to reah the paok (45,000 m 2 ) where they were turne out. During turnout perio eah paok ha from 30 to 70 horses. Turnout in April to June was performe in the same way. Exerise regimen hanges were implemente at the beginning of Table 1 Daily feeing sheule of a group of Brazilian army horses Time Commerial Conentrate (Kg) Bermua Hay (Kg) 4:00 AM 2 e 11:00 AM e 2 1:00 PM 2 e 4:00 PM e 2 5:30 PM 2 e Conentrate was ompose of rimpe oats, linsee meal, soybean meal, wheat bran, grass hay, molasses, rake orn, soybean oil, probiotis, mineral an vitamin premix, alium arbonate, an salt. the month, when appliable. In February, horses were exerise at a light intensity (Table 2) an were walke for 60 minutes three times a week, an aoring to National Researh Counil (NRC; 2007), this ativity may inlue trot an anter too. This ativity was part of the normal military ativity where horses neee to be prepare to walk in the ity. This level of ativity also prepares the soliers who never roe before. In Marh, ativity was inrease to a moerate intensity an horses were walke an trotte for 90 minutes three times a week, an aoring to NRC (2007), it inlues some walking, mostly trotting, an some anter uring 3 to 5 hours a week. During this month, soliers were traine to trot an horses were prepare to work on guar ativities in the ity. In the months of April, May, an June, horses were turne out beause the Guar Regiment was involve in other ativities. In July, horses were worke again at a moerate intensity (NRC, 2007), most of the time at walk an about 40% of the time at trot (Table 2) for 5 ays a week for 90 minutes. This exerise intensity was part of the normal guar ativities for horses that worke on guar rouns in the ity. Ativities were stoppe in November, when horses were turne out again. The exerise regimen was applie to all horses Bloo Sampling, Hanling, an Analysis Bloo samples were ollete between the eighth an 19th of eah month. Horses ha free aess to water uring the night an their last meal of grain (Table 1) was onsume by 6:00 PM the night before sampling. Monthly jugular bloo samples were rawn between 2:00 an 3:30 AM into heparinize vautainer tubes an fluorize tubes (BD Vautainer evauate bloo olletion tube, Fisher HealthCare, Chiago, IL) before the morning feeing at 4:30 AM. Tubes were plae on ie water an entrifuge at 3,000g for 10 minutes within 30 minutes of olletion. Plasma was remove, maintaine at 4 C for 4 hours until frozen at 20 C for further analysis. Insulin was etermine by raioimmunoassay (Coat-A- Count Insulin, Diagnosti Prouts, Los Angeles, CA) previously valiate for equine insulin [16]. The proxy for insulin sensitivity was alulate from basal plasma onentrations of insulin (mu/l) as follows: RISQI ¼ 1/Obasal insulin onentration ¼ basal insulin onentration 0.5 [10] Statistis Insulin an insulin sensitivity (RISQI) were ompare aross months with exerise intensity as a main effet by analysis of variane with repeate measures. For statistial analysis, plasma insulin an RISQI values were logtransforme beause of their non-normal istribution. Multiple omparisons were mae by Fisher least square means analysis. Subjet age, gener, an weight were inlue as ovariates in the analysis of variane. Spearman rank orrelations were alulate with age, BCS, an weight to etermine their effet on insulin an insulin sensitivity. Bimonthly hay sample nutritional ontent was analyze by linear trens analysis. All statistial analyses were performe using SAS software (SAS Institute, Cary, NC). Results are presente as means an SEM.

3 S.P. Turner et al. / Journal of Equine Veterinary Siene 31 (2011) Table 2 Monthly physial ativity of a group of Brazilian army horses Physial ativity Months Type Duration Frequeny Intensity November 2006eJanuary 2007 Turnout 8 hours 4 ays per week Turnout February Walk 60 minutes 3 ays per week Light Turnout 6 hours 3 ays per week Marh Walk an trot 90 minutes 3 ays week Moerate Turnout 8 hours 2 ays week April, May, an June Turnout 8 hours 4 ays week Turnout JulyeOtober Walk an trot 90 minutes 5 ays week Moerate NovembereJanuary 2008 Turnout 8 hours 4 ays per week Turnout 3. Results 3.1. Horse Demographis an Sampling Days Of the 31 horses, 14 were sample uring all 9 months an 17 were sample on 8 of 9 months. Age of the horses varie between 6 an 23 years ( years), with no age ifferene between months. Horses were not teste for Cushing isease, but no horses emonstrate linial signs of the isease. Gener of the horses onsiste of 68% gelings an 32% mares, with no ifferene between months. Weights an BCS i not vary among months (P ¼.99). Average weight was kg (range: 305 to 513 kg) an BCS was (range: 3 to 7). Bloo samples were ollete on February 8, Marh 8, April 10, May 10, June 15, July 10, August 15, September 19, an Otober Diet an Management Fee analysis is shown in Table 3. No variation was foun in the bimonthly onentrate or hay omposition an ata for eah were poole. Horses were fe their onentrate in stalls an they generally onsume all onentrate an hay offere. Hay onsumption was not ontrolle iniviually uring turnout months when fe in the afternoon. Grass intake was onsiere insignifiant beause the paok onsiste primarily of irt. Conentrate intake base on the horses boy weight was 1.40% 0.01% (range: 1.16% to 1.96%) an average hay intake 0.95% 0.01% as fe (range: 0.78% to 1.31%) Weight, BCS, Conentrate Perentage, an Gener BCS was negatively assoiate (r ¼ 0.21, P ¼.030) with RISQI, whereas it was positively assoiate with insulin onentrations (r ¼.16) an age (r ¼.17) (P <.010). Conentrate perentage fe base on boy weight (range: 1.16% to 1.96%) was not a signifiant fator in any analysis Physial Ativity The intra-assay an interassay oeffiient of variation of insulin upliate samples were 8% an 11%, respetively. The majority of the results, however, were within the lowest level of the stanar urve of the insulin assay. Overall, basal fasting insulin onentration was higher (P ¼.013) in months where horses reeive only turnout (log estimate ¼ ; insulin mean ¼ mu/ml) than in months where horses unerwent light exerise (log estimate ¼ ; insulin mean ¼ mu/ml) an lowest (P <.001) with moerate exerise intensity (log estimate ¼ ; insulin mean ¼ mu/ml) (Fig. 1). Insulin sensitivity was lower (P ¼.013) in turnout horses (log estimate ¼ ; RISQI mean ¼ ) than lightly exerise horses (log estimate ¼ ; RISQI mean ¼ ) an also lower (P <.001) than moerately exerise horses (log estimate ¼ ; RISQI mean ¼ ) (Fig. 2). Insulin sensitivity was higher (P <.001) in moerately exerise horses as ompare with lightly exerise horses. Insulin an RISQI were not signifiantly ifferent between the months of February, Marh, an April (Figs. 1, 2). Insulin an RISQI were lower in Marh, as ompare with May an June (P <.014), an higher than July, August, September, an Otober (P <.008) (Figs. 1, 2). Although there was only a tren for a ifferene in insulin an RISQI between February an Marh (P ¼.067), an Marh an April (P ¼.068), overall turnout insulin an RISQI were ifferent (P.013) from light exerise intensity (Figs. 1, 2). Table 3 Means an SEM for the poole hay an grain hemial analysis Feestuff DM Energy (Mal/kg) CP Ash Fat NDF ADF NFC a CHO-H b CHO-F R Hay Grain DM, ry matter; CP, rue protein; NDF, neutral etergent fiber; ADF, ai etergent fiber; NFC, nonfibrous arbohyrate; CHO-H, hyrolyzable arbohyrate; CHO-F R, rapily fermentable arbohyrate. a Nonstrutural arbohyrate, NFC ¼ 100 ash CP fat NDF. b Analyze iretly using extration with hot water an amylolyti enzymes [13]. CHO-F R ¼ NFC CHO-H.

4 648 S.P. Turner et al. / Journal of Equine Veterinary Siene 31 (2011) Insulin ( µu/ml) ab b 4. Disussion ab a a Feb M a r h April M a y J un e J uly Aug S ep O t Light ativity Moerate ativity Turn out Fig. 1. Monthly means an SEM for insulin. Bars with ifferent supersripts iffer (P <.050). This stuy foun that exerise intensity an affet insulin sensitivity in horses, an that turnout alone may not be suffiient to improve insulin sensitivity. The majority of the horses in this stuy were young an not obese, thus lowering the risk of insulin resistane. Horses were fe about 0.95% of their boy weight as forage, whih is lose to the minimum reommene forage intake [17]. Consequently, onentrate intake was high, whih an preispose to insulin resistane [2,4,18,19]. However, lower CHO-H (166 g/kg DM) an CHO-F R (106 g/kg DM) ontent of the onentrate fe in this stuy oul have reue the likelihoo of the horses to evelop insulin resistane, as has been shown to our with fat- an fiber-base iets [14]. The low insulin values reporte in this stuy are also likely to be beause of fasting before the sampling. Although supplie fee i vary uring the stuy, BCS an weights i not. It is possible, however, that uring turnout months, horses i not onsume all 2 kg of offere hay, whih is estimate to ontain about 3.9 Mal, thus reuing the energy intake. Despite low insulin onentrations, this stuy showe that exerise was assoiate with lower irulating insulin onentrations, suggesting a positive effet on insulin sensitivity. In human beings, regular moerate physial ativity (40% to 55% of their peak oxygen onsumption) has been shown to improve insulin sensitivity [20,21]. In horses, exerise has also been shown to improve insulin sensitivity in previously inative obese an lean mares [1], untraine Stanarbre horses [3], an lean Stanarbre horses [2]. However, in the lean Stanarbres [2], insulin i not reflet this ifferene in insulin sensitivity, possibly beause of small sample size an iniviual variability in basal values. Insulin sensitivity returne to pretraining levels in mares within 9 ays after training stoppe [1], iniating that training shoul be ontinuous to maintain improve sensitivity. In obese horses that are likely to be insulin-resistant, fee restrition in aition to exerise might be neessary to improve insulin sensitivity; however, one stuy foun that 4 weeks of low intensity exerise followe by 4 weeks of higher intensity exerise i not improve insulin sensitivity, although it reue obesity moerately [18]. In the urrent stuy, moerate exerise improve insulin sensitivity within 15 ays (Fig. 2; July [turnout] as ompare with August [moerate exerise]). Differenes in insulin onentration between light exerise (Fig. 1; February) an moerate exerise (Marh) were not evient. Insulin onentration in Marh uring moerate exerise one three times a week was still higher than insulin onentration in the months of moerate exerise intensity in July to Otober, where horses were exerise 5 ays aweek(fig. 1). Exerise at moerate intensity may nee to be frequent to improve insulin sensitivity. The results of the urrent stuy suggest that ontinuous moerate exerise performe at least 5 ays a week an reue the risk of insulin resistane in horses in as little as 15 ays. From the urrent stuy, it is not lear for how long insulin sensitivity will remain the same if exerise is not unertaken. Although horses reeiving turnout were not ompare with horses with no turnout in this stuy, it is unlikely that horses turne out on pasture will signifiantly inrease their heart rate an oxygen onsumption to prompt metaboli aaptations. 5. Conlusions Moerate exerise intensity performe 5 ays a week seems to benefit insulin sensitivity, even in insulinsensitive horses. Turnout may not be suffiient to improve insulin sensitivity, but an exerise regime omparable with light or moerate intensity exerise, as performe in the stuy (e.g. walking horses for 60 minutes five times a week), may be reommene as a management strategy to improve insulin sensitivity. This shoul be teste in an at-risk population of obese horses. RISQI ab b ab a a Feb Marh April May June July Aug Sep Ot Light ativity Moerate ativity Turn out Fig. 2. Monthly means an SEM for reiproal of the square root of insulin. Bars with ifferent supersripts iffer (P <.050). Aknowlegments This stuy was fune by Funação o Amparo a Pesquisa o Estao o Rio e Janeiro an by the Laboratory for Equine Performane at the Feeral Rural University of Rio e Janeiro (Laboratório e Avaliação o esempenho e equinos a Universiae Feeral Rural o Rio e Janeiro). Speial thanks to Dr Raquel Gouvea os Santos from Centro e Desenvolvimento a Tenologia Nulear an to Dr Patríia Vieira, veterianarian from the Cavalry Regiment.

5 S.P. Turner et al. / Journal of Equine Veterinary Siene 31 (2011) Referenes [1] Powell DM, Reey DR, Sessions DR, Fitzgeral BP. Effet of shortterm exerise training on insulin sensitivity in obese an lean mares. Equine Vet J Suppl 2002;34:81-4. [2] Stewart-Hunt L, Geor RJ, MCutheon LJ. Effets of short-term training in insulin sensitivity an skeletal musle gluose metabolism in Stanarbre horses. Equine Vet J Suppl 2006;36: [3] Graaf-Roelfsema E, Ginneken ME, van Brena E, Winjberg ID, Keizer HA, van er Kolk JH. The effet of long-term exerise on gluose metabolism an peripheral insulin sensitivity in Stanarbre horses. Equine Vet J Suppl 2006;36: [4] Hoffman RM, Boston RC, Stefanovsky D, Kronfel DS, Harris PA. Obesity an iet affet gluose ynamis an insulin sensitivity in Thoroughbre gelings. J Anim Si 2003;81: [5] Pratt SE, Geor RJ, Spriet LL, MCutheon LJ. Time ourse of insulin sensitivity an skeletal musle glyogen synthase ativity after a single bout of exerise in horses. J Appl Physiol 2007;103: [6] Treiber KH, Hess TM, Kronfel DS, Boston RC, Geor RJ, Freire M, et al. Gluose ynamis uring exerise: ietary energy soures affet minimal moel parameters in traine Arabian gelings uring enurane exerise. Equine Vet J Suppl 2006;36: [7] Tinker MK, White NA, Lessar P, Thather CD, Pelzer KD, Davis B, et al. Prospetive stuy of equine oli iniene an mortality. Equine Vet J 1997;29: [8] Hess TM, Kronfel DS, Cranell KE, Walron JN, Treiber KH, Staniar WB, et al. Fat aaptation affets insulin sensitivity an elimination uring an 80 km enurane rie. Pfereheilkune 2007;23: [9] Bailey SR, Habershon-Buther JL, Ranson KJ, Elliott J, Menzies- Gow NJ. Hypertension an insulin resistane in a mixe bree population of ponies preispose to laminitis. Am J Vet Res 2008;69: [10] Treiber KH, Kronfel DS, Hess TM, Boston RC, Harris PA. Use of proxies an referene quintiles obtaine from minimal moel analysis for etermination of insulin sensitivity an panreati betaell responsiveness in horses. Am J Vet Res 2005;66: [11] Henneke D, Potter GD, Kreier JL, Yeates BF. Relationship between onition sore, physial measurements an boy fat perentage in mares. Equine Vet J 1983;15: [12] Costa LAP. Manual e hippologia. Rio e Janeiro, Brazil: Bibliotea o Exérito; p [13] Smith D. Removing an analyzing total nonstrutural arbohyrates from plant tissue. Maison, WI: University of Wisonsin; p Researh Report R2107. [14] Hoffman RM, Wilson JA, Kronfel DS, Cooper WL. Hyrolysable arbohyrates in pasture, hay, an horse fees: iret assay an seasonal variation. J Anim Si 2001;79: [15] NRC, National Researh Counil. Nutrient requirements of horses. 6th rev e. Washington, DC: National Aaemy Press; p [16] Freestone J, Wolfsheimer KJ, Kamerling SG, Churh G, Hamra C, Bangwell C. Exerise inue hormonal an metaboli hanges in Thoroughbre horses: effets of onitioning an aepromazine. Equine Vet J 1991;23: [17] Meyer H, Raike S, Kienzle E, Wilke S, Kleffken D, Illenseer M. Investigations on preileal igestion of starh form grain, potato an manio in horses. ZBL Vet Me A 1996;42: [18] Carter RA, MCutheon J, Valle E, Meilan EN, Geor RJ. Effets of exerise training on aiposity, insulin sensitivity, plasma hormone an lipi onentrations in overweight or obese, insulin-resistant horses. Am J Vet Res 2010;71: [19] Chatterton NJ, Harrison PA, Bennett JH, Asay KH. Carbohyrate partitioning in 185 aessions of graminea grown uner warm an ool temperatures. J Plant Physiol 1989;143: [20] Bassuk SS, Manson JE. Epiemiologial eviene for the role of physial ativity in reuing risk of type 2 iabetes an ariovasular isease. J Appl Physiol 2005;99: [21] Houmar JA, Tanner CJ, Slentz CA, Dusha BD, MCartney JS, Kraus WE. Effet of the volume an intensity of exerise training on insulin sensitivity. J Appl Physiol 2004;96:101-6.

EFFECT OF DIFFERENT METHODS OF PRESERVATION ON THE QUALITY OF CATTLE AND GOAT MEAT. Abstract

Bang. J. Anim. Si. 2009, 38(1&2) : 86 91 ISSN 0003-3588 EFFECT OF DIFFERENT METHODS OF PRESERVATION ON THE QUALITY OF CATTLE AND GOAT MEAT S. Bin. Faisal, S. Akhter 1 and M. M. Hossain Abstrat The study