||J. Haisan, Y. Inabu, W. Shi, and M. Oba, Effects of pre- and postpartum dietary starch content on productivity, plasma energy metabolites, and serum inflammation indicators of dairy cows, https://doi.org/10.3168/jds.2020-19611, 2021.03, The objective of this study was to evaluate the effects of the starch content of pre- and postpartum diets on productivity, plasma energy metabolites, and serum markers of inflammation of dairy cows during the calving transition period. Eighty-eight primiparous and multiparous cows were randomly assigned to pre- and postpartum dietary treatments balanced for parity and pretrial body condition score at d 28 ± 3 before expected calving date. Cows were fed either a control [Control; 14.0% starch, dry matter (DM) basis] or high-starch (High; 26.1% starch, DM basis) prepartum diet commencing 28 ± 3 d before expected calving date. Following calving, cows were fed either a high-fiber (HF; 33.8% neutral detergent fiber, 25.1% starch, DM basis) or high-starch (HS; 27.2% neutral detergent fiber, 32.8% starch, DM basis) postpartum diet for the first 20 ± 2 d following calving. Cows fed the High prepartum diet had greater DM intake (12.4 vs. 10.2 kg/d), plasma concentrations of insulin (1.72 vs. 14.2 ng/mL), glucose (68.1 vs. 65.0 mg/dL), and glucagon-like peptide-2 (0.41 vs. 0.32 ng/mL) before parturition, but increased plasma free fatty acid concentration (452 vs. 363 µEq/L) and milk fat yield (1.64 vs. 1.48 kg/d) after parturition. Cows fed the HS postpartum diet had lower plasma free fatty acid (372 vs. 442 µEq/L) and serum haptoglobin (0.46 vs. 0.70 mg/mL) concentrations over a 3-wk period after calving. In addition, there was a tendency for interaction between prepartum and postpartum diets for milk yield, where feeding the HS postpartum diet increased milk yield compared with the HF diet for cows fed the Control prepartum diet (40.8 vs. 37.9 kg/d) but not for cows fed the High prepartum diet. These results suggest that management efforts to minimize the change in diet fermentability during the calving transition by feeding the High prepartum diet, the HF postpartum diet, or both did not increase productivity of dairy cows but increased fat mobilization after calving. Our findings also suggest that feeding high-starch postpartum diets can decrease fat mobilization and serum indicators of systemic inflammation and increase milk production even with the transition from a low-starch prepartum diet..
||Yudai Inabu, Hiroshi Yamamoto, Haruki Yamano, Yutaka Taguchi, Shunnosuke Okada, Tetsuji Etoh, Yuji Shiotsuka, Ryoichi Fujino, Hideyuki Takahashi, Glucagon-like peptide 2 (GLP-2) in bovine colostrum and transition milk, Heliyon, DOI:https://doi.org/10.1016/j.heliyon.2021.e07046, 2021.05, Bovine colostrum contains growth factors, cytokines, hormones, and enzymes, which have important roles in stimulating gastrointestinal development of neonatal calves. In the present study, we measured the concentration of glucagon-like peptide 2 (GLP-2), one of the gut-derived peptides secreted from intestinal L-cells, in colostrum and transition milk of Japanese black cattle. All colostrum samples were collected within 24 h after calving (d 0) and transition milk was collected at 24, 48 and 72 h relative to the time at colostrum sampling (d 1, d 2 and d 3, respectively). Concentrations of GLP-2 in colostrum were 5.53 ± 1.07 ng/mL on average (range = 0.94–9.60 ng/mL) and decreased from d 0 to 3 (P < 0.01). Furthermore, concentrations of GLP-2 in colostrum and transition milk were quadratically decreased with the elapsed time from parturition until colostrum sampling (R2 = 0.48, P < 0.01). Our results show for the first time that GLP-2 is present in bovine colostrum and transition milk and that concentrations decreased with elapsed time from parturition..
||Yutaka Taguchi, Yudai Inabu, Koki Hayasaki, Noriyuki Maeda, Yoshiro Kanmera, Seiji Yamasaki, Noboru Ota, Kenji Mukawa, Arisa Tsuboi, Hirokuni Miyamoto, Tetsuji Etoh, Yuji Shiotsuka, Ryoichi Fujino, Christopher D McMahon, Hideyuki Takahashi, Effects of feeding high volumes of milk replacer on reproductive performance and on concentrations of metabolites and hormones in blood of Japanese black heifer calves, Animal Science Journal, DOI: 10.1111/asj.13505 , Anim Sci J. 2021;92:e13505., 2021.01, We evaluated the effects of feeding high volumes of milk replacer on growth and reproductive performances in Japanese black heifers. Fifty-one heifers were fed milk replacer at 9 L/d for 60 d (9L*60d; n = 18) or 41 d (9L*41d; n = 15), or at 7 L/d for 40 d (7L*40d; n = 18). Artificial insemination (AI) was performed on heifers with ≥ 270 kg body weight and ≥ 116 cm body height at 300 d of age. The age at the first AI was 0.35 mo later for 7L*40d than the other groups (P < 0.01). However, age at calving did not differ among treatments (22.1 mo). The interval from the first AI to pregnancy tended to be ~2 mo longer for the 9L*60d than the other groups (P = 0.07). Our results showed that feeding high volumes of milk replacer may reduce the age at calving via an improved rate of growth. In addition, we propose that feeding a maximum of 7 L milk replacer for 40 d may be the most appropriate rearing regime because the success of pregnancy per AI may be reduced in calves fed a maximum of 9 L for 41 and 60 d..
||Y Inabu, J Haisan, M Oba, T Sugino, Effects of feeding a moderate- or high-energy close-up diet to cows on response of newborn calves to milk replacer feeding and intravenous injection of glucagon-like peptide 1, Domestic Animal Endocrinology, 74:106528. doi: 10.1016/j.domaniend.2020.106528., 2020.07, In this study, we investigated the effects of feeding a moderate- or high-energy close-up diet to close-up cows on response of newborn calves to intravenously (i.v.) injected glucagon-like peptide 1 (GLP-1). Newborn Holstein heifer calves (n = 37) from cows fed with a moderate-energy [M, 1.54 Mcal/kg of dry matter (DM) NEl; 14% starch; n = 17] or high-energy (H, 1.63 Mcal/kg of DM NEl; 26% starch; n = 20) diet in the last 28 d prepartum were assigned to one of two treatment groups, which were i.v. injected with saline (MC and HC, n = 9 and 10, respectively) or GLP-1 solution at 1.0 μg/kg BW (MG and HG, n = 8 and 10, respectively) immediately after milk replacer (MR; 26% CP, 16% fat) feeding. Blood samples were obtained through a jugular vein catheter at -10, 0, 10, 20, 30, 40, 50, 60, 90, and 120 min relative to MR feeding at 2, 10, and 20 d after birth, and plasma glucose, insulin, and GLP-1 concentrations were measured. Plasma GLP-1 concentration tended to increase starting from 30 min after MR feeding in the MC relative to the HC group at 10 (0.77 ng/mL vs 0.69 ng/mL for MC and HC, respectively; P = 0.10) and 20 d after birth (0.47 ng/mL vs 0.35 ng/mL for MC and HC, respectively; P = 0.07). Plasma glucose and insulin concentrations after MR feeding did not differ between MC and HC groups at 2 and 20 d after birth but were higher (P < 0.05) in MC (158 mg/dL and 3.64 ng/mL for glucose and insulin, respectively) than in HC (143 mg/dL and 2.46 ng/mL for glucose and insulin, respectively) calves at 10 d after birth. The elevation in plasma glucose concentration after MR feeding was suppressed by direct glucose-lowering action of i.v. injected GLP-1 at 2, 10, and 20 d after birth in M and H calves; this direct glucose-lowering action by GLP-1 was greater (P < 0.05) for H than for M calves at 20 d after birth. These results indicate that feeding a high-energy close-up diet to cows affects glucose status in their female offspring via suppression of postprandial plasma concentrations of GLP-1 and insulin as well as the alteration in the glucose-lowering action of GLP-1 after feeding depending on the day after birth..
||M. Elsabagh, Y. Inabu, T. Obitsu, T. Sugino, Response of plasma glucagon-like peptide-2 to feeding pattern and intraruminal administration of volatile fatty acids in sheep, Domestic Animal Endocrinology, 10.1016/j.domaniend.2017.03.001, 60, 31-41, 2017.07, Glucagon-like peptide-2 (GLP-2), a gut peptide secreted by enteroendocrine L cells, has recently been identified as a key regulator of intestinal growth and absorptive function in ruminants. However, reports on GLP-2 secretion are few, and more information regarding its secretion dynamics is needed. In this study, two experiments were conducted to elucidate the daily rhythm of GLP-2 secretion in response to feeding regimen and to investigate the effect of volatile fatty acids (VFA) on GLP-2 release in sheep. In experiment 1, blood samples were collected over 3 d from 4 Suffolk mature wethers adapted to a maintenance diet fed once daily; day 1 sampling was preceded by 24 h of fasting to reach steady state. On days 1 and 3, samples were collected every 10 min from 11:00 to 14:00 on both days and then every 1 h until 00:00 on day 1 only; feed was offered at 12:00. On day 2, feed was withheld, and sampling was performed every hour from 01:00 to 00:00. In experiment 2, 5 Suffolk mature wethers were assigned to 5 treatment groups of intraruminal administration of saline, acetate, propionate, butyrate, or VFA mix (acetate, propionate, and butyrate in a ratio of 65:20:15) in a 5 × 5 Latin square design. Blood samples were collected at 0, 1.5, 3, 6, 9, 12, 15, 20, 25, 30, 40, 50, 60, 90, and 120 min relative to the beginning of administration at 12:00. In both experiments, plasma GLP-2, glucagon-like peptide-1 (GLP-1), glucose, insulin, and β-hydroxy butyric acid (BHBA) levels were measured. In experiment 1, incremental area under the curve was greater (P < 0.05) post-feeding than pre-feeding on days 1 and 3 for GLP-2 and tended to be greater (P < 0.1) on day 1 for GLP-1. Plasma insulin, glucose, and BHBA levels increased (P < 0.05) on day 1 post-feeding. Plasma GLP-2 was poorly correlated with GLP-1 but positively correlated with insulin, glucose, and BHBA. In experiment 2, administration of butyrate and VFA mix remarkably increased plasma GLP-2 (P = 0.05) and BHBA (P < 0.0001) levels compared with those in other treatments. Plasma GLP-1 levels were higher with butyrate administration compared with those in the saline, acetate, and VFA mix (P = 0.019). Propionate administration increased plasma glucose (P = 0.013) and insulin (P = 0.053) levels. Thus, our data confirmed that GLP-2 release is responsive to feeding and might be promoted by BHBA produced by the rumen epithelial metabolism of butyrate. Further molecular- and cellular-level studies are needed to determine the role of butyrate as a signaling molecule for GLP-2 release..
||A. Saegusa, K. Inouchi, M. Ueno, Y. Inabu, S. Koike, T. Sugino, M. Oba, Effects of partial replacement of corn grain with lactose in calf starters on ruminal fermentation and growth performance, Journal of Dairy Science, 10.3168/jds.2017-12616, 100, 8, 6177-6186, 2017.08, The objective of this study was to evaluate effects of partial replacement of dry ground corn with lactose in calf starters on dry matter intake, growth rate, ruminal pH, and volatile fatty acid profile. Sixty Holstein bull calves were raised on a high plane of nutrition program until 55 d of age. Calves were fed texturized calf starters containing 30.1% steam-flaked grains and lactose at 0 (control), 5, or 10% (n = 20 for each treatment) on a dry matter basis. All calves were fed treatment calf starters ad libitum from d 7 and kleingrass hay from d 35. Ruminal pH was measured continuously immediately after weaning (d 55–62) for 15 calves (n = 5 per treatment), and 3 wk after weaning (d 77 to 80) for the other 45 calves (n = 15 per treatment). Dry matter intake, growth performance, and ruminal pH variables were not affected by treatment. However, according to Spearman's correlation coefficient (rs) analyses, lactose intake was positively correlated with dairy minimum ruminal pH (rs = 0.306) for the data collected from d 77 to 80. Similarly, hay intake was not affected by treatment, but positively correlated with daily mean (rs = 0.338) and maximum ruminal pH (rs = 0.408) and negatively correlated with duration pH <5.8 (rs = −0.329) and area pH <5.8 (rs = −0.325), indicating that the variation in hay intake among animals might have masked treatment effects on ruminal pH. Ruminal molar ratio of acetate was higher (45.2 vs. 40.6%), and that of propionate was lower in 10% lactose than control (35.3 vs. 40.2%) for ruminal fluid collected on d 80; however, molar ratio of butyrate was not affected by treatment. These results indicate that lactose inclusion in calf starters up to 10% of dry matter might not affect dry matter intake and growth performance of calves, but that greater lactose and hay intake might be associated with higher ruminal pH..
||Y. Inabu, A. Saegusa, K. Inouchi, S. Koike, M. Oba, T. Sugino, Plasma concentrations of glucagon-like peptide 1 and 2 in calves fed calf starters containing lactose, Journal of Dairy Science, 10.3168/jds.2017-12910, 100, 11, 9361-9371, 2017.11, The objective of this study was to evaluate the effects of lactose inclusion in calf starters on plasma glucagon-like peptide (GLP)-1 and GLP-2 concentrations and gastrointestinal tract development in calves. Holstein bull calves (n = 45) were raised on an intensified nursing program using milk replacer containing 28.0% CP and 15.0% fat, and were fed a texturized calf starter containing 0 (control), 5.0 (LAC5), or 10.0% (LAC10; n = 15 for each treatment) lactose on a DM basis. Lactose was included in the starter by partially replacing dry ground corn in pelleted portion of the starter. All calf starters were formulated with 23.1% CP. The ethanol-soluble carbohydrate concentrations of the control, LAC5, and LAC10 starters were 7.3, 12.3, and 16.8% on a DM basis, respectively. Starch concentrations of the control, LAC5, and LAC10 starters were 29.7, 27.0, and 21.4% on a DM basis, respectively. All calves were fed treatment calf starters ad libitum. Blood samples were obtained weekly from 1 to 11 wk of age, and used to measure plasma GLP-1, GLP-2, and insulin concentrations, serum β-hydroxybutyrate (BHB) concentration, and blood glucose concentration. At 80 d of age, calves were euthanized, and weights of the reticulorumen, omasum, abomasum, small intestine, and large intestine tissue were measured. Serum BHB concentration was higher for calves fed the LAC10 (171 μmol/L) starter than for those fed the control (151 μmol/L) and LAC5 (145 μmol/L) starters. Plasma GLP-1 and GLP-2 concentrations did not differ between treatments. However, relative to the baseline (1 wk of age), the plasma GLP-1 concentration was higher for the LAC10 (125.9%) than for the LAC5 (68.2%) and control (36.8%), and for the LAC5 than for the control (36.8%). Moreover, similar differences between treatments were observed for GLP-2 concentration relative to the baseline (88.2, 76.9, and 74.9% for LAC10, LAC5, and control treatments, respectively). The serum BHB concentration was positively correlated with the plasma GLP-1 concentration (r = 0.428). Furthermore, the plasma GLP-1 concentration was positively correlated with the insulin concentration (r = 0.793). The weights of the reticulorumen, omasum, abomasum, small intestine, and large intestine were not affected by the treatments. In conclusion, inclusion of lactose in calf starters resulted in higher plasma GLP-1 and GLP-2 concentrations, and BHB might be associated with higher plasma GLP-1 concentration..
||Y. Inabu, A. Fischer, Y. Song, L. L. Guan, M. Oba, M. A. Steele, T. Sugino, Short communication
The effect of delayed colostrum feeding on plasma concentrations of glucagon-like peptide 1 and 2 in newborn calves, Journal of Dairy Science, 10.3168/jds.2018-14412, 101, 7, 6627-6631, 2018.07, Glucagon-like peptide (GLP)-1 is involved in glucose homeostasis via its role in stimulating insulin secretion, whereas GLP-2 increases mucosal growth of the small intestine. To our knowledge, the effect of delayed colostrum feeding on plasma GLP-1 and GLP-2 in neonatal calves has not been evaluated. To investigate the effect of delayed colostrum feeding on plasma concentrations of GLP-1 and GLP-2 in newborn calves, we randomly assigned 27 Holstein bull calves to 1 of 3 treatment groups: those fed colostrum within 1 h after birth (control), 6 h after birth (6H), and 12 h after birth (12H; n = 9 for each treatment). Blood samples were obtained before the colostrum feeding and every 3 h after each colostrum feeding for a 36-h period, and plasma concentrations of GLP-1, GLP-2, insulin, and glucose were measured. Plasma GLP-1 concentration at 12 h after colostrum feeding was lower in 12H than in control calves. In addition, plasma insulin concentration was lower in the 6H and 12H calves than in the controls. Plasma glucose and GLP-2 concentrations were, however, not affected by treatment. These results indicate that delayed colostrum feeding can decrease plasma GLP-1 and insulin concentrations without affecting glucose or GLP-2 concentration..
||Bayissa Hatew, Yudai Inabu, Toshihisa Sugino, Michael Steele, Effects of pulse-dose ruminal infusion of butyrate on plasma glucagon-like peptide 1 and 2 concentrations in dairy calves, Journal of Dairy Science, 10.3168/jds.2018-15578, 102, 3, 2254-2265, 2019.03, Feeding of butyrate was found to have a positive effects in enhancing gut development and improving growth performance of calves. Equally, glucagon-like peptide 1 and 2 (GLP-1 and GLP-2), secreted from gastrointestinal L-cells in response to nutrient intake, were found to play a significant role in regulating blood glucose homeostasis and improving gut health. However, limited information is available about the relationship between butyrate and release of GLP-1 and GLP-2 in dairy calves. The objective of this study was to evaluate the effects of a pulse-dose ruminal infusion of butyrate on plasma GLP-1 and GLP-2 concentrations in dairy calves. Five ruminally cannulated mature Holstein bull calves (7.2 ± 0.10 mo, and 330 ± 16.0 kg of body weight; mean ± standard deviation) were used in a 5 × 5 Latin square with 4-d periods. On d 1 of each period at 0800 h, calves were ruminally infused with 1 of 5 treatments: 0 (saline), 0.3, 0.6, 0.9, and 1.2 g of butyrate per kg of body weight. Before butyrate infusion, calves were not offered feed overnight, and sequential blood and rumen fluid samples were taken before and after infusion on d 1 of each period. Ruminal butyrate and total volatile fatty acid concentrations increased linearly (2.65, 12.19, 20.99, 30.19, and 36.30; 23.68, 33.07, 40.94, 51.13, and 56.31 µmol/mL, for butyrate and total volatile fatty acids, respectively) in a dose-dependent manner, whereas propionate and isobutyrate increased quadratically. Ruminal and plasma butyrate, β-hydroxybutyrate, GLP-1, GLP-2, insulin, and glucose concentrations were all affected by treatment, time (except GLP-2), and interaction of treatment with time (except GLP-1). The area under the curve (AUC) summarized at different time points relative to the baseline (AUC30, AUC60, AUC120, and AUC240) for ruminal and plasma butyrate, and BHB, increased linearly with the dose of butyrate infused. However, AUC30, AUC60, AUC120, and AUC240 for plasma GLP-2 concentration were affected in a cubic manner unlike the linear effect on AUC30 and AUC60 for GLP-1. Plasma GLP-2 was not correlated with plasma butyrate (r = 0.16), GLP-1 (r = 0.03), or BHB (r = −0.05). This findings suggest that pulse-dosing of butyrate slightly increased both GLP-1 and GLP-2 concentrations at specific time points and this might be promoted by direct or indirect effect of butyrate on the intestinal L-cells..
||Y. Inabu, J. Pyo, S. Pletts, L. L. Guan, M. A. Steele, T. Sugino, Effect of extended colostrum feeding on plasma glucagon-like peptide-1 concentration in newborn calves, Journal of Dairy Science, 10.3168/jds.2018-15616, 102, 5, 4619-4627, 2019.05, Glucagon-like peptide-1 (GLP-1) plays a role in the regulation of glucose homeostasis via the stimulation of insulin secretion. The objective of this study was to evaluate the effect of extended colostrum feeding on plasma concentration of GLP-1. Holstein bull calves (n = 27) were fed pooled colostrum at 7.5% of birth body weight at 2 h after birth and then fed mature milk (M), a 50:50 mixture of pooled colostrum and milk (CM), or pooled colostrum (C; n = 9 for each treatment) at 5% of birth body weight at 12 h after birth and every 12 h thereafter until 72 h after birth. Blood samples were obtained before (1 and 2 h after birth) and after (until 72 h after birth; 42 time points) the first colostrum feeding, and plasma concentrations of glucose, insulin, and GLP-1 were measured. Data were analyzed by ANOVA of JMP 13 (SAS Institute Inc., Cary, NC) with treatment, time, and treatment × time interaction as fixed effects. Treatment × time interaction was observed for plasma insulin and glucose concentrations, which were mainly the result of lower concentrations from 1 to 2 d after birth for C compared with M. Conversely, on d 3 after birth, the difference between treatments was not observed for insulin and glucose. For the entire experimental period, plasma GLP-1 concentration was higher for C (2.25 ng/mL) compared with M (1.41 ng/mL) and tended to be higher compared with CM (1.58 ng/mL). A treatment × time interaction was observed for GLP-1, but unlike glucose and insulin, this was mainly the result of higher concentrations from 54 to 72 h after birth (on d 3 after birth) for C compared with M or CM. Postprandial plasma concentration of glucose was not correlated with that of GLP-1 but was positively correlated with that of insulin for the 4-h period after feeding on d 1 (r = 0.30) and d 3 after birth (r = 0.33). Postprandial plasma concentration of GLP-1 was positively correlated with that of insulin for the 4-h period after feeding on d 3 after birth (r = 0.20). These results indicate that extended colostrum feeding may increase plasma GLP-1 concentrations, especially 3 d after birth, but further study is necessary to determine the effect on plasma insulin and glucose concentrations..
||J. Haisan, Y. Inabu, W. Shi, M. Oba, Effects of feeding a high- or moderate-starch prepartum diet to cows on newborn dairy heifer calf responses to intravenous glucose tolerance tests early in life, Journal of Dairy Science, 10.3168/jds.2018-16226, 102, 10, 8931-8940, 2019.10, The objective of this study was to evaluate the effect of feeding a prepartum diet with a high or moderate starch content on growth and insulin sensitivity of female offspring early in life. Thirty-eight Holstein heifer calves were born to dams fed either a high-starch (26% starch on a DM basis, HI; n = 20) or moderate-starch (14% starch on a DM basis, MOD; n = 18) prepartum diet commencing at 28 ± 3 d before expected parturition date. Following birth, all calves were housed individually and fed three 2-L meals of colostrum within the first 24 h of life and offered 10 L/d of milk replacer (26% CP, 18% fat, mixed to 130 g/L). Body weight of calves was measured at birth and on d 2 (after colostrum feeding but before milk feeding), 10 ± 2, and 20 ± 2. A glucose tolerance test was performed at a minimum of 6 h after their last colostrum or milk meal to evaluate insulin sensitivity on d 2, 10 ± 2 and 20 ± 2. Body weight did not differ throughout between HI and MOD calves; however, calves born to primiparous dams were smaller compared with those born to multiparous dams. Glucose or insulin concentrations were not different before the glucose tolerance test. Following the glucose tolerance test, maximum glucose concentrations were not different between treatments at any time point. However, HI calves had greater insulin area under the curve, and HI calves had greater maximum insulin concentrations on d 2. Glucose or insulin clearance rates were not different nor was the calculated insulin sensitivity index between treatments. These findings suggest that feeding a HI prepartum diet may reduce some insulin sensitivity indicators of female offspring early in life..
||Yudai Inabu, Kyotaro Murayama, Katsutoshi Inouchi, Toshihisa Sugino, The effect of tributyrin supplementation to milk replacer on plasma glucagon-like peptide 2 concentrations in pre-weaning calves, Animal Science Journal, 10.1111/asj.13262, 90, 9, 1185-1192, 2019.09, The objective of this study was to evaluate the effect of tributyrin (TB) supplementation to milk replacer (MR) on performance, health, and blood concentrations of metabolite and glucagon-like peptide (GLP-2) in pre-weaning calves. Twenty Holstein heifer calves were raised on an intensified nursing program using MR supplemented with either palm oil (CON) or TB (TB) at 0.3% (as fed basis) for 7 weeks starting 1 week after birth. Calves were fed a calf starter and kleingrass from the beginning of the study. Blood samples were obtained weekly to measure blood glucose, serum β-hydroxybutyric acid (BHBA), insulin-like growth factor 1 (IGF-1), and plasma GLP-2 concentrations. Starter DMI and metabolizable energy (ME) intake were lower in TB calves at 46, 47, from 49 to 55 days after birth compared with the CON calves. However, any growth parameters were not affected by TB treatment. Blood glucose, serum BHBA, and IGF-1 concentrations were not affected by TB supplementation. On the other hand, mean plasma GLP-2 concentration among whole experimental period was higher for TB (0.60 ng/ml) compared with CON (0.41 ng/ml). In conclusion, feeding MR supplemented with TB increases plasma GLP-2 concentration, which might counterbalance the growth performance of TB calves despite the decreased ME intake..