Toshihisa Sugino

Effects of feeding a calf starter on molecular adaptations in the ruminal epithelium and liver of Holstein dairy calves

The objective of this study was to elucidate the effect of feeding a calf starter on the volatile fatty acid (VFA) profile in the rumen and on expression of genes involved in epithelial intracellular pH regulation, butyrate metabolism, and hepatic urea cycle during the weaning transition. Twenty Holstein bull calves were fed either milk replacer and hay (MR) or milk replacer, hay, and a commercial texturized calf starter (MR+S) in a randomized complete block design. All calves were fed 750 g/d of milk replacer as the basal diet. Calves on the MR+S treatment were also fed starter ad libitum, and the energy intake of calves within blocks was maintained by supplementing the MR group with extra milk replacer that was equivalent to the energy intake from calf starter. Calves were killed 3 d after they consumed 680 g/d of calf starter for 3 consecutive days. Calves fed MR+S had higher VFA concentrations in the rumen (99.1±8.1 vs. 64.6±8.6 mM) and a higher molar proportion of butyrate (15.6±1.7 vs. 7.9±1.9%) than calves fed MR. Relative abundance of mRNA for monocarboxylate transporter isoform 1 was higher (1.45 vs. 0.53), and that of Na(+)/H(+) exchanger isoform 3 (0.37 vs. 0.82) and 3-hydroxy-3-methylglutaryl synthase isoform 1 (0.40 vs. 0.94) lower for the MR+S treatment compared with the MR treatment. In the liver, relative mRNA abundances of argininosuccinate synthetase isoform 1 (2.67 vs. 1.56), argininosuccinate lyase (1.44 vs. 0.99), and arginase isoform 1 (3.21 vs. 1.74) were greater for MR+S than for MR calves. Calf starter consumption appeared to increase fermentation in the rumen and affected expression of genes involved in cholesterol synthesis and intracellular pH regulation in ruminal epithelium, and those involved in urea cycle in the liver.

Comparisons of insulin related parameters in commercial-type chicks: Evidence for insulin resistance in broiler chicks

The aim of this study is to elucidate whether insulin acts differentially within the central nervous system (CNS) of two types of commercial chicks to control ingestive behavior. Male layer and broiler chicks (4-day-old) were intracerebroventricularly (ICV) injected with saline or insulin under satiated and starved conditions. Feed intake was measured at 30, 60 and 120 min after treatment. Secondly, blood and hypothalamus were collected from both chick types under ad libitum feeding and fasting for 24 h. Plasma insulin concentration was measured by time-resolved fluoro-immunoassay. Hypothalamic insulin receptor mRNA expression levels were measured by quantitative RT-PCR. The ICV injection of insulin significantly inhibited feed consumption in layer chicks when compared with saline (P<0.05), but not broiler chicks (P>0.1). Plasma insulin concentration of both chick types significantly decreased following 24 h of fasting, while insulin concentrations in the broiler chicks were significantly higher compared to the layers fed under ad libitum conditions. Hypothalamic insulin receptor mRNA expression levels were significantly lower (P<0.05) in broiler chicks than in layer ones under ad libitum feeding. Feed deprivation significantly decreased insulin receptor mRNA levels in layer chicks (P<0.01), but not in broiler chicks (P>0.1). Moreover, plasma insulin concentrations correlated negatively with hypothalamic insulin receptor protein expression in the two types of chicks fed ad libitum (P<0.05). These results suggest that insulin resistance exists in the CNS of broiler chicks, possibly due to persistent hyperinsulinemia, which results in a down-regulation of CNS insulin receptor expression compared to that in layer chicks.

Effects of starch content of calf starter on growth and rumen pH in Holstein calves during the weaning transition.

The objective of this study was to evaluate the effects of substituting high fiber byproducts for dry ground corn in calf starter on growth and rumen pH during the weaning transition. Holstein bull calves were raised on an intensified nursing program using milk replacer containing 26% CP and 18% fat. Calves were fed a texturized calf starter containing either dry ground corn at 18.8% of dry matter (DM; CRN), beet pulp replacing dry ground corn at 10.2% dietary DM (BP), or triticale dried distillers grains with solubles replacing dry ground corn and high-protein feedstuffs at 18.6% of dietary DM (DDGS) in the pellet; treatment calf starters differed only in the pellet portion. Starch concentrations of CRN, BP, and DDGS were 35.3, 33.4, and 31.4%, respectively. After a calf consumed 2.50 kg of starter for 3 consecutive days, a small ruminant rumen pH data logger was inserted orally and rumen pH was measured continuously for 4d. Calves were then killed and rumen fluid was sampled to determine volatile fatty acid profile. No difference was found in overall average daily gain or growth rates of hip height, withers height, and heart girth. During the weaning transition, rate of increase in calf starter intake was greater for calves fed DDGS compared with those fed CRN (87.7 vs. 77.5 g/d), but lower for calves fed BP compared with CRN (68.1 vs. 77.5 g/d). The area under pH 5.8 (470 vs. 295 min × pH/d) or pH 5.2 (72.7 vs. 16.4 min × pH/d) was greater for calves fed DDGS than those fed CRN. Rumen pH profile was not affected by BP treatment compared with CRN, but calves fed BP tended to have greater water intake than those fed CRN (6.6 vs. 5.8 L/d). Volatile fatty acid profile was not affected by treatment with the exception of molar proportion of butyrate, which tended to be lower for calves fed BP compared with those fed CRN (15.0 vs. 16.6%). Hay intake was positively correlated to mean rumen pH for calves used in this study (r=0.48). Decreasing dietary starch concentration did not mitigate rumen acidosis in calves during weaning transition, and low rumen pH did not adversely affect growth during the weaning transition.

Effects of glucose and amino acids on ghrelin secretion in sheep

Two experiments were conducted to elucidate the effects of post-ruminal administration of starch and casein (Exp. 1), plasma amino acids concentrations (Exp. 2), and plasma glucose and insulin concentrations (Exp. 2) on plasma ghrelin concentrations in sheep. In Exp. 1, plasma ghrelin concentrations were determined by four infusion treatments (water, cornstarch, casein and cornstarch plus casein) in four wethers. Abomasal infusion of casein increased plasma alpha-amino N (AAN) concentrations. Infusion of starch or casein alone did not affect plasma ghrelin concentrations, but starch plus casein infusion increased plasma levels of ghrelin, glucose and AAN. In Exp 2, we investigated the effects of saline or amino acids on ghrelin secretion in four wethers. Two hours after the initiation of saline or amino acid infusion into the jugular vein, glucose was also continuously infused to investigate the effects of blood glucose and insulin by hyper-glycemic clump on plasma ghrelin concentrations. Infusion of amino acids alone raised plasma levels of ghrelin, but the higher plasma glucose and insulin concentrations had no effect on plasma ghrelin concentrations. These results suggest that high plasma levels of amino acids can stimulate ghrelin secretion, but glucose and insulin do not affect ghrelin secretion in sheep.

Effects of high ambient temperature on urea‐nitrogen recycling in lactating dairy cows

Effects of exposure to hot environment on urea metabolism were studied in lactating Holstein cows. Four cows were fed ad libitum a total mixed ration and housed in a temperature-controlled chamber at constant moderate (18°C) or high (28°C) ambient temperatures in a cross-over design. Urea nitrogen (N) kinetics was measured by determining urea isotopomer in urine after single injection of [(15) N(2) ]urea into the jugular vein. Both dry matter intake and milk yield were decreased under high ambient temperature. Intakes of total N and digestible N were decreased under high ambient temperature but urinary urea-N excretion was increased. The ratio of urea-N production to digestible N was increased, whereas the proportion of gut urea-N entry to urea-N production tended to be decreased under high ambient temperature. Neither return to the ornithine cycle, anabolic use nor fecal excretion of urea-N recycled to the gut was affected by ambient temperature. Under high ambient temperature, renal clearance of plasma urea was not affected but the gut clearance was decreased. Increase of urea-N production and reduction of gut urea-N entry, in relative terms, were associated with increased urinary urea-N excretion of lactating dairy cows in higher thermal environments.

Ghrelin differentially modulates the GH secretory response to GHRH between the fed and fasted states in sheep

The effect of energy balance on the growth hormone (GH) secretory responsiveness to growth hormone-releasing hormone (GHRH) has not been determined in ruminant animals. Therefore, we examined the effects of intravenous injections of 0, 3.3, and 6.6 microg ghrelin/kg body weight (BW), with and without GHRH at 0.25 microg/kg BW, on GH secretory responsiveness in both the fed and fasted sheep. The injections were carried out at 48 h (Fasting state) and 3h (Satiety state) after feeding. Blood samples were taken every 10 minutes, from 30 minutes before to 120 minutes after the injection. Low (3.3 microg/kg BW) and high (6.6 microg/kg BW) doses of ghrelin stimulated GH secretion significantly (P<.05) greater in the Satiety state than in the Fasting state. Growth hormone-releasing hormone plus both doses of ghrelin stimulated GH secretion significantly (P<.05) greater in the Satiety state than in the Fasting state. Ghrelin and GHRH exerted a synergistic effect in the Satiety state, but not in the Fasting state. Plasma ghrelin levels were maintained significantly (P<.05) greater in the Fasting state than in the Satiety state except the temporal increases after ghrelin administration. Plasma free fatty acid (FFA) concentrations were significantly (P<.01) greater in the Fasting state than in the Satiety state. In conclusion, the present study has demonstrated for the first time that ghrelin differentially modulates GH secretory response to GHRH according to feeding states in ruminant animals.

Ingestion of medium chain fatty acids by lactating dairy cows increases concentrations of plasma ghrelin.

The purpose of this study was to elucidate the effects of medium-chain fatty acids (MCFAs) on plasma ghrelin concentration in lactating dairy cows. Five early-lactating Holstein cows were randomly assigned to 2 dietary treatments in a crossover design with 2-wk periods. Treatments consisted of diets supplemented or not (control) with calcium salts of MCFAs (MCFA-Ca; 1.5% dry matter). Plasma hormone and metabolite concentrations in blood samples taken from the jugular vein were measured on the morning of feeding on day 14 of each period. Dry matter intake, milk protein, and lactose content of cows fed the MCFA-Ca diet were decreased compared with controls, but with no change in milk yield. Plasma ghrelin concentrations were higher in cows fed the MCFA-Ca diet; however, no significant effect was found on glucagon-like peptide-1 concentrations in plasma. Plasma insulin concentrations decreased, but plasma glucagon concentrations remained unchanged in cows fed the MCFA-Ca diet. The concentrations of nonesterified FAs, total cholesterol, and β-hydroxybutyrate in plasma increased in these cows. In conclusion, dietary MCFAs increase the plasma ghrelin concentrations in lactating dairy cows.

Plasma concentrations and effects of glucagon-like peptide-1 (7-36) amide in calves before and after weaning.

Glucagon-like peptide-1 (7-36) amide (GLP-1), secreted by the small intestine, has insulinotropic and glucose-lowering action. Basal plasma GLP-1 concentrations were measured in calves around the weaning period, the effect of short-chain fatty acids (SCFA) on plasma GLP-1 concentrations was examined, and the effects of GLP-1 administration on plasma insulin, glucagon, and glucose concentrations were measured. Thirteen Holstein bull calves were fed whole milk and solid feed and weaned at 7 wk of age. Preprandial plasma samples were obtained from 5 calves once a week from week 0 to 13 to measure basal concentrations of plasma GLP-1 and insulin (experiment 1). Four calves were intravenously administered with a mixed solution of SCFA (2.4 mmol/kg body weight [BW]) in week 2 and 11 to measure plasma GLP-1 concentrations (experiment 2). Another 4 calves were intravenously injected with GLP-1 (1.0 μg/kg BW) to elucidate the response of plasma insulin, glucagon, and glucose concentrations in week 1, 2, 4, 6, 7, 9, 11, and 13 (experiment 3). In experiment 1, age and weaning did not affect preprandial basal concentrations of plasma GLP-1 throughout the experimental period. Preprandial insulin concentrations increased after weaning (P < 0.05), and GLP-1 and insulin were more strongly correlated postweaning than preweaning. In experiment 2, intravenous treatment with SCFA increased plasma GLP-1 concentrations in both week 2 and 11 (P < 0.05.) In experiment 3, intravenous GLP-1 treatment decreased plasma glucose concentrations throughout the experiment (P < 0.05), but increased plasma insulin concentrations only after weaning (P < 0.05). Treatment with GLP-1 did not affect plasma glucagon concentrations, regardless of age. These results indicate that preprandial basal concentrations of plasma GLP-1 in calves are not changed by weaning, but SCFA stimulate GLP-1 secretion. The insulinotropic action of GLP-1 is detected only after weaning, but the glucose-lowering action of GLP-1 is not affected by weaning.

A preliminarily study for predicting body weight and milk properties in lactating Holstein cows using a three-dimensional camera system

Digital imaging has been applied to assess body weight and fatness in livestock.We examine low priced 3D camera for estimating cow body weight and milk properties.Six geodesic line (GL) lengths were computed using back posture 3D object of cow.A similar determination of body condition with standard method is possible. Since manual body condition scoring has been widely utilized as an indirect and subjective method to estimate energy reserves of dairy cattle, image analysis has been increasingly researched for use on large farms as an objective and effective measuring instrument for the estimation of body condition score (BCS) and body weight (BW). Recent advances in the technological development of the three-dimensional (3D) cameras may provide innovative feed management tools for dairy farms. The objective of the present study was to evaluate the feasibility of a 3D camera systems in measuring the back posture of lactating Holstein dairy cows to predict the BCS, BW, milk yield (MY), milk fat (MF) and milk protein (MP). The BCSs for eight cows were recorded by two trained observers using a 5-point scale, and other variables were obtained using an automatic milking system during the lactation. Back posture measurements of dairy cows were conducted using the ASUS Xtion Pro sensor. Six geodesic line (GL) lengths were computed using the 3D objects of each cow based on the positions of the right and left hook bones (GLhh), right and left thurl bones (GLtt), right and left pin bones (GLpp), hook and thurl bones (GLht), hook and pin bones (GLhp), and coccygeal ligament (GLcl). In the principal component analysis (PCA), GL, GLpp, and GLcl had the greatest contribution to principal component values (PCV) 1, 2, and 3, respectively, and these three PCVs described 0.887 of the cumulative contribution ratio. Good correlations were found between the observed and predicted values of BCS (R2=0.74), BW (0.80), MY (0.62), MF (0.62), and MP (0.53) based on linear regression equations using the GLs as explanatory variables and parity (1, 2, and >3) as a fixed effect. These results demonstrate that the 3D cameras could represent an innovative tool for estimating body condition and milk properties.

Ghrelin differentially modulates glucose-induced insulin secretion according to feeding status in sheep

The present study was conducted to investigate roles of ghrelin in glucose-induced insulin secretion in fasting- and meal-fed state in sheep. Castrated Suffolk rams were fed a maintenance diet of alfalfa hay cubes once a day. Hyperglycemic clamp (HGC) was carried out to examine glucose-induced insulin response from 48 to 53 h (fasting state) and from 3 to 8 h (meal-fed state) after feeding in Experiment 1 and 2 respectively. Total dose of 70 nmol/kg body weight of D-Lys3-GHRP6, a GH secretagogue receptor 1a (GHS-R1a) antagonist, was intravenously administered at 0, 60, and 120 min after the commencement of HGC. In the fasting state, the ghrelin antagonist significantly (P < 0.01) enhanced glucose-induced insulin secretion. In the meal-fed state, i.v. administration of synthetic ovine ghrelin (0.04 microg/kg body weight per min during HGC) significantly (P < 0.05) enhanced glucose-induced insulin secretion. d-Lys3-GHRP6 treatment suppressed ghrelin-induced enhancement of the insulin secretion. In conclusion, ghrelin has an inhibitory and stimulatory role in glucose-induced insulin secretion via GHS-R1a in fasting- and meal-fed state respectively.