Influence of feeding a fish oil-containing diet to mature, overweight dogs: Effects on lipid metabolites, postprandial glycaemia and body weight.

The objective of this study was to determine the effect of feeding a fish oil (FO)-containing diet on lipid and protein metabolism, postprandial glycaemia and body weight (BW) of mature, overweight dogs. Seven female dogs were randomly assigned to one of two isonitrogenous and isocaloric diets, control (CO) or FO (FO), in a crossover design. Experimental periods were 69 day, separated by a washout period of 30 day. At the beginning of the experiment, and at 30 and 60 day of feeding the experimental diets, the dogs were infused with D-glucose (2 g/kg BW) through an intravenous catheter. Blood samples were collected for 3 hr to perform a glucose tolerance test. Nitrogen balance measurements began at 06:30 on d 63 of each experimental period and ended at 06:30 on d 69. On d 66 of each period, a single dose (7.5 mg/kg) of 15 N-glycine was administered orally for determination of protein turnover. Incremental area under the curve and glucose concentration at peak did not differ between treatments or among sampling days within treatment. Glucose half-life tended to decrease (p < .10) in the FO treatment on day 30 when compared to baseline (day 0). ß-hydroxybutyrate, non-esterified fatty acid (NEFA) and triglycerides did not differ within or between treatments. Cholesterol decreased (p < .05) on the FO treatment on day 30, 60 and 69 when compared to day 0. High-density lipoprotein (HDL) decreased (p < .05) in the FO treatment on day 69 when compared to day 0. Body weight, food intake, faecal excretion, DM and N digestibilities, N balance and protein turnover were not different between diets. Overall, FO-containing diet decreases cholesterol in mature overweight dogs; however, further research is warranted to verify the effects of FO on glucose metabolism.

Glucagon-like peptide-2 (GLP-2) increases small intestinal blood flow and mucosal growth in ruminating calves.

Glucagon-like peptide-2 (GLP-2) increases small intestinal mass and blood flow in nonruminants but its effect in ruminants is unknown. Eight Holstein calves with an ultrasonic flow probe around the superior mesenteric artery and catheters in the carotid artery and mesenteric vein were paired by age and randomly assigned to treatment of a control (0.5% of BSA in saline; n=4) or GLP-2 (50 µg/kg of body weight of bovine GLP-2 in BSA; n=4) given subcutaneously every 12h for 10 d. Blood flow was measured on d 0 (acute) and d 10 (chronic) and included 3 periods: baseline (saline infusion), treatment (infusion of BSA or 1,000 pmol of GLP-2/kg of body weight per h), and recovery (saline infusion). On d 11, calves were killed 2h after injection of 5-bromo-2'-deoxyuridine (BrdU). Gastrointestinal tissues were weighed and epithelial samples were obtained to determine villus height, crypt depth, and BrdU staining. Infusion of GLP-2 increased superior mesenteric artery blood flow to 175% of baseline on d 0 but to only 137% of baseline after chronic treatment. Compared with that of the control, GLP-2 increased small intestinal mass by 24% by increasing epithelial mass in the jejunum and ileum. Additionally, GLP-2 increased villus height, crypt depth, and BrdU-labeling in small intestinal segments. These results demonstrate that GLP-2 induces similar increases in small intestinal blood flow and growth in ruminants to those observed in nonruminants. Furthermore, GLP-2 increases small intestinal blood flow in ruminants but this response is attenuated after 10 d of GLP-2 administration. In cattle, GLP-2 may be an important hormone in the regulation of intestinal blood flow and epithelial growth.

Ruminal and abomasal starch hydrolysate infusions selectively decrease the expression of cationic amino acid transporter mRNA by small intestinal epithelia of forage-fed beef steers.

Although cationic amino acids (CAA) are considered essential to maximize optimal growth of cattle, transporters responsible for CAA absorption by bovine small intestinal epithelia have not been described. This study was conducted to test 2 hypotheses: 1) the duodenal, jejunal, and ileal epithelia of beef cattle differentially express 7 mRNA associated with 4 mammalian amino acid (AA) transport activities: y(+) (CAT1), B(0,+) (ATB(0,+)), b(0,+) (b(0,+)AT and rBAT), and y(+)L (y(+)LAT1, y(+)LAT2, and 4F2hc), and 2) the expression of these mRNA is responsive to small intestinal luminal supply of AA substrates (derived from ruminal microbes) or glucose-derived energy (from starch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (body weight = 260 +/- 17 kg) fed an alfalfa cube-based diet at 1.33 x net energy for maintenance requirement were assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control); ruminal SH and abomasal water infusion; and ruminal water and abomasal SH infusion. The dosage of SH infusion amounted to 20% of metabolizable energy intake. After 14 or 16 d of infusion, steers were slaughtered, duodenal, jejunal, and ileal epithelia were harvested, and total RNA was extracted. The relative amounts of mRNA expressed by epithelia were quantified using real-time reverse transcription-PCR. All 7 mRNA species were expressed by the epithelium from each region, but their abundance differed among the regions. Specifically, duodenal expression of CAT1 and ATB(0,+) mRNA was greater than jejunal or ileal expression; ileal expression of b(0,+)AT, rBAT, and y(+)LAT1 mRNA was greater than jejunal or duodenal expression, whereas the expression of y(+)LAT2 and 4F2hc mRNA did not differ among the 3 epithelia. With regard to SH infusion effect, ruminal infusion down-regulated or tended to down-regulate the jejunal expression of CAT1, rBAT, y(+)LAT2, and 4F2hc mRNA. Abomasal infusion down-regulated the jejunal expression of y(+)LAT2 mRNA and tended to down-regulate the jejunal expression of 4F2hc mRNA. This study characterized the pattern of CAA transporter mRNA expressed by growing beef cattle fed an alfalfa-based diet. Moreover, this study demonstrated that increasing the luminal supply of microbe-derived AA (by ruminal supplementation of SH) results in a reduced capacity of apical and basolateral membrane to transport of CAA, whereas increasing luminal glucose supply (by abomasal supplementation of SH) reduces only the basolateral transport capacity, assuming that CAA transporter mRNA content represents functional capacity.

Basal expression of nucleoside transporter mRNA differs among small intestinal epithelia of beef steers and is differentially altered by ruminal or abomasal infusion of starch hydrolysate.

In ruminants, microbial-derived nucleic acids are a major source of N and are absorbed as nucleosides by small intestinal epithelia. Although the biochemical activities of 2 nucleoside transport systems have been described for cattle, little is known regarding the regulation of their gene expression. This study was conducted to test 2 hypotheses: (1) the small intestinal epithelia of beef cattle differentially express mRNA for 3 concentrative (CNT1, 2, 3) and 2 equilibrative (ENT1, 2) nucleoside transporters (NT), and (2) expression of these NT is responsive to small intestine luminal supply of rumen-derived microbes (hence, nucleosides), energy (cornstarch hydrolysate, SH), or both. Eighteen ruminally and abomasally catheterized Angus steers (260 +/- 17 kg of BW) were fed an alfalfa cube-based diet at 1.33x NE(m) requirement. Six steers in each of 3 periods were blocked by BW (heavy vs. light). Within each block, 3 steers were randomly assigned to 3 treatments (n = 6): ruminal and abomasal water infusion (control), ruminal SH infusion/abomasal water infusion, or ruminal water infusion/abomasal SH infusion. The dosage of SH infusion amounted to 20% of ME intake. After a 14-or 16-d infusion period, steers were slaughtered, and duodenal, jejunal, and ileal epithelia were harvested for total RNA extraction and the relative amounts of mRNA expressed were determined using real-time RT-PCR quantification methodologies. All 5 NT mRNA were found expressed by each epithelium, but their abundance differed among epithelia. Specifically, jejunal expression of all 5 NT mRNA was higher than that by the ileum, whereas jejunal expression of CNT1, CNT3, and ENT1 mRNA was higher, or tended to be higher, than duodenal expression. Duodenal expression of CNT2, CNT3, and ENT2 mRNA was higher than ileal expression. With regard to SH infusion treatments, ruminal infusion increased duodenal expression of CNT3 (67%), ENT1 (51%), and ENT2 (39%) mRNA and ileal expression of CNT3 (210%) and ENT2 (65%) mRNA. Abomasal infusion increased (54%) ileal expression of ENT2 mRNA and tended to increase (50%) jejunal ENT2 mRNA expression. This study has uniquely characterized the pattern of NT mRNA expression by growing beef cattle and found that the mRNA abundance for CNT3, ENT1, and ENT2 in small intestinal epithelia can be increased by increasing the luminal supply of nucleotides (CNT3, ENT1, ENT2) or glucose (ENT2).

Conception risk of beef cattle after fixed-time artificial insemination using either SexedUltra™ 4M sex-sorted semen or conventional semen.

The objective of this study was to compare conception rates of female beef cattle inseminated at a fixed-time with either conventional (CON) or SexedUltra™ sex-sorted (SU) semen. Treatments included CON or SU with two sires represented within each treatment. Cows (n = 316) and heifers (n = 78) from six locations were randomly assigned treatment. Ovulation was synchronized in all females using the industry-standard 7-d CO-Synch + controlled internal drug release (CIDR) protocol (100 µg GnRH + CIDR [1.38 g progesterone] on d 0, 25 mg PGF2α at CIDR removal on d 7, and 100 µg GnRH on d 10, 54 h (heifers) or 60 h (cows) after CIDR removal). Estrotect™ estrous detection aids were applied at CIDR removal and patch activation was recorded at insemination. Animals were assumed estrual if greater than 50% of the patch coating was removed. The results from this study indicated no main effects of treatment (P = 0.82), sire (P = 0.64), or age (P=0.8) on AI conception rates. Additionally, there were not significant interactions between sire and treatment (P=0.19) or age and treatment (P=0.29). There was however, a significant (P=0.0005) effect of estrous expression on conception rates. Conception rate for estrual females (62.8%) was greater (p=0.0001) than non-estrual females (38.7%) at FTAI regardless of treatment. Furthermore, the conception rates were similar (P = 0.61) between conventional (61.9%) and sex-sorted semen (63.8%) when estrus was expressed prior to FTAI. Larger studies are warranted to determine appropriate timing of insemination with sex-sorted semen in FTAI protocols to maximize pregnancy potential.

A novel objective chute score interacts with monensin to affect growth of receiving cattle.

Growth in cattle may be related to animal temperament via alterations in intake or feed conversion. However, temperament is ill-defined, and different temperament measures may relate differently to production traits or interact with dietary factors in their effects. To examine relationships between diet, temperament, growth, and health, 160 crossbred steers (262 ± 22 kg) were used in a 56-d RCBD experiment with a 2 × 2 × 2 factorial treatment structure with 5 pens/treatment. Steers were pen fed a corn silage-based diet with or without monensin (41.9 g/t DM), ad libitum. Temperament treatments (assigned on d -7) were exit velocity (EV; slow vs. fast) and objective chute score (OCS; low vs. high), a novel temperament measure, representing the CV of weights collected at 5 measures/s for 10 s while an animal's head was restrained in a chute. Both were measured on d -7, 0, 14, 28, 55, and 56. Subjective chute scores (SCS; visual estimates of animal activity obtained simultaneously with OCS measures) were measured on d -7 and d 56. Jugular blood samples from d 28 were analyzed for antibody response to leptospirosis vaccine and NEFA concentrations. No monensin × OCS × EV interactions were detected ( ≥ 0.11). There was a positive correlation between SCS and OCS ( < 0.01; = 0.57). Changes in OCS and EV across the duration of the study differed among treatments (treatment × day, < 0.10) and indicated that initial measures may be better proxies of growth than average measures. There were no interactions between EV and OCS ( ≥ 0.15) for any response variable and no interactions among treatments ( ≥ 0.31), nor main effects of temperament factors ( ≥ 0.12) for DMI (%BW). Monensin decreased DMI ( < 0.01) similarly across all levels of EV and OCS. Gains and G:F responses to monensin depended on OCS ( < 0.10) but not EV ( ≥ 0.80). Gain was reduced ( < 0.10) by monensin with low, but not high, OCS, and G:F was increased ( < 0.10) by monensin on high, but not low, OCS. Gain during the second 4 wk was lesser ( = 0.04) in fast, compared with slow, EV animals. Results provide novel indications that certain temperament measures can interact with dietary manipulation to influence animal performance.

Relationships of a novel objective chute score and exit velocity with growth performance of receiving cattle.

Animals with excitable temperaments often have decreased gains that have been associated with decreased intake and efficiency. Different temperament measures probably measure different specific underlying traits. Commonly used temperament measures include both objective and subjective measures. Subjective measures present potential difficulties for making across-study comparisons and thus for generalizing quantitative relationships. One objective of this experiment was to evaluate 2 related, but different, measures associated with temperament, where 1 measure is a new, objective measurement based on the common subjective chute score measures. Also, there is reason to believe that RDP requirements of animals may vary with temperament. To examine the relationships between temperament measures and nutrient use, 192 crossbred steers were used in a 58-d randomized complete block design experiment. Temperament treatments (assigned prior to d 1) were chute exit velocity (EV; slow vs. fast) and objective chute score (WSD; low vs. high), a novel temperament measure that was the SD of weights collected at 5 Hz for 10 s while an animal was restrained in a chute with its head caught. Both were measured on d -8, 1, 2, 16, 30, 56, and 58, where d 1 was the day that animals were allotted to treatment groups and began receiving experimental diets. Steers were fed a diet with 1 of 3 RDP levels (75%, 105%, and 120% of RDP requirements). There were no main effects or interactions with RDP ( ≥ 0.12); thus, it was removed from the statistical model for subsequent analyses. There were no interactions between EV and WSD ( ≥ 0.11). Slow EV animals had greater ADG ( = 0.02) and DMI ( ≤ 0.09) than fast EV animals, but there was no effect of EV on G:F ( > 0.14). For d 0 to 58, high WSD animals had greater DMI ( ≤ 0.09) than low WSD animals but no difference in ADG ( = 0.23), whereas low WSD animals tended to have increased G:F ( = 0.11). Results of this study give additional confirmation that EV is associated with DMI and growth and provide evidence that a novel measure of behavior, WSD, is also related to growth, independently of EV. Because WSD and EV appear to measure different underlying behavioral traits, use of both measures may improve our ability to discriminate among temperament categories for growing cattle.

Direct-fed microbials containing lactate-producing bacteria influence ruminal fermentation but not lactate utilization in steers fed a high-concentrate diet.

Direct-fed microbials (DFM) have been shown to improve gain and growth efficiency and also modulate ruminal fermentation. In Exp. 1,72 beef steers were used to compare a lactate-producing bacterial (LAB) DFM consisting primarily of Lactobacillus acidophilus and Enterococcus faecium,and a lactate-producing and lactate-utilizing (LAB/LU) DFM consisting primarily of L. acidophilus and Propionibacterium both fed at 10(9) cfu/d. Steers were fed a corn-based finishing diet for 153 d and then slaughtered for collection of carcass characteristics. In Exp. 2, 12 ruminally cannulated steers were fed acorn-based finishing diet and treated with 10(9) cfu/d of LAB DFM. Rumen fluid was sampled on d 14 and 28 over a 12-h period. Steers were ruminally dosed with a 2-L solution of neutralized DL-lactate (0.56 M)and Cr-EDTA (13.22 M) 3 h postfeeding on d 15 and 29. Ruminal samples were collected at 10- and 20-minintervals for the first and second hour postdosing. No differences (P ≥ 0.14) between control (CON) and LAB for DMI, ADG, growth efficiency, or carcass characteristics were observed. Dry matter intake was greater (P = 0.04) for LAB/LU than LAB from d 0 to 28 but did not differ (P ≥ 0.29) thereafter. Average daily gain was greater (P = 0.04) and efficiency tended(P = 0.06) to be greater for LAB than LAB/LU over the entire 153 d. In Exp. 2, total VFA concentration and molar proportions of butyrate were unaffected(P ≥ 0.24). Molar proportions of acetate exhibited a DFM by hour interaction (P = 0.04); however, on average, molar proportion of acetate was 4.4% greater for DFM. Conversely, DFM did not affect the molar proportion of propionate (P = 0.39). On average,molar proportions of propionate tended to increase(P = 0.07), and acetate tended to decrease (P = 0.07)across days. Mean daily ruminal pH was similar for CON on d 14 and 28, whereas mean pH increased from d 14 to 28 for DFM (DFM × day; P = 0.08).Minimum pH remained unchanged for CON over time but increased from d 14 to 2 for DFM (DFM × day;P = 0.10). Maximum pH decreased from d 14 to 28 in CON but increased over time with DFM (DFM × day;P = 0.05). DL- and L-lactate utilization were unaffected by DFM (P ≥ 0.33) or day (P ≥ 0.50). Although the LAB DFM did not impact growth performance, itd id modulate ruminal fermentation, as evidenced by shifts in ruminal VFA profile and pH; however, DFM did not appear to influence ruminal lactate utilization.

Effect of intake on fasting heat production, respiratory quotient and plasma metabolites measured using the washed rumen technique.

The objective was to investigate the effect of intake before fasting on concentrations of metabolites and hormones, respiratory quotient (RQ) and fasting heat production (HP) using the washed rumen technique and to compare these values with those from the fed state. Six Holstein steers (360±22 kg) were maintained at 21°C and fed three different energy intakes within a replicated 3×3 Latin square design with 21-day periods. Steers were fed alfalfa cubes to provide 1.0, 1.5 and 2.0×NEm during 19 days of each experimental period. Steers were placed in individual metabolism stalls fitted with indirect calorimetry head-boxes on day 20 of each experimental period (FED steers) and fed their normal meal. On day 21 of each period the reticulorumen was emptied, washed and refilled with ruminal buffer (NaCl=96; NaHCO3=24; KHCO3=30; K2HPO4=2; CaCl2=1.5; MgCl2=1.5 mmol/kg of buffer) aerated with 75% N2 and 25% CO2 before introduction to the rumen (steers were not fed; WASHED steers). Each gas exchange was measured over 24 h. HP for 1.0, 1.5 and 2.0×NEm were 479, 597 and 714 kJ/daykg0.75 (s.e.m. =16), respectively. The plateau RQ was 0.756, 0.824 and 0.860 for the 1.0, 1.5 and 2.0×NEm intakes for the FED steers, respectively. After rumen washing, fasting HP was 331, 359 and 400 kJ/daykg0.75 (s.e.m.=13) for 1.0, 1.5, and 2.0×NEm intakes before fasting, respectively. The RQ for WASHED rumen steers was 0.717, 0.710 and 0.719, respectively. Cortisol and ß-hydroxybutyrate concentrations in WASHED rumen steers did not exceed threshold levels for severe energy deficit and stress as can be induced from prolonged fasting. This study demonstrates that a fasting state can be emulated using the washed rumen technique, minimizing the time required as opposed to traditional fasting methodologies, without causing a severe energy deficit and stress.

Influence of fiber fermentability on nutrient digestion in the dog.

Eight mature dogs (17.2 +/- 0.2 kg) surgically fitted with ileal T-cannulas were used in a replicated 4-x-4 Latin-square-design experiment to evaluate nutrient disappearance at the terminal ileum and through the digestive tract. Two fiber types, cellulose, a crystalline, slowly fermented fiber, and pectin, a soluble, rapidly fermented fiber, were fed in different increments, and the effects on nutrient availability were assessed. Treatments included 1) 100% cellulose, 2) 66% cellulose and 33% pectin, 3) 66% pectin and 33% cellulose, and 4) 100% pectin. Fiber was added at 10% of diet dry matter (DM). Diets were fed at 100% of ME for maintenance and offered at 0730 and 1730 h. All periods were 21 d, which included 3 d of diet transition and 7 d of adaptation. Daily DM intake was 210 +/- 5 g. Total tract and large-intestine DM digestibility increased linearly (P < 0.01) with increased pectin. These changes in DM digestion were largely the result of changes in fiber digestion. Fermentation of total dietary fiber in the large intestine went from less than zero to 39% of ileal flow (linear, P < 0.01). Total-tract crude-protein digestibility decreased linearly (P < 0.01) with increased pectin. This study demonstrated that fiber fermentability significantly affects digestion in the dog. Increasing fermentable fiber increased the digestion of DM and energy. However, increased fiber fermentability inversely affects crude protein digestibility. The lower crude-protein digestibility could be attributed to larger microbial protein excretion as a result of greater fermentation of pectin versus cellulose.

Effects of cysteamine on pulsatile growth hormone release and plasma insulin concentrations in sheep.

The effects of cysteamine (CSH; 0, 50, or 100 mg/kg BW), a somatostatin depleting agent, on growth hormone (GH) and insulin (INS) secretion were studied in sheep (Ovis aries). Cysteamine was administered as a single intragastric bolus on day 0 (0900). Jugular blood samples were collected at 15-min (GH) and 2-hr (INS) intervals over an 8-hr period (1100-1900) on day 0, 3, and 7. Intragastric administration of CSH at 50 mg/kg BW augmented (quadratic, P = .04) mean plasma GH concentration, with the greatest response occurring on day 3. Baseline GH concentrations were elevated in wethers dosed with 50 mg/kg BW CSH on day 3, whereas wethers dosed with 100 mg/kg BW CSH had lower baseline GH concentrations on day 0 (CSH x day interaction, P = .02). Cysteamine administration increased GH pulse amplitude (quadratic, P = .15), with the greatest magnigtude of change occurring with 50 mg/kg BW CSH on day 0 and 3. Frequency of GH pulses was increased (quadratic, P = .10) following CSH treatment. Administration of 100 mg/kg BW CSH augmented plasma INS on day 0 (CSH x day interaction, P = .09). These findings indicate that CSH alters GH and INS secretion in a dose-dependent and temporal manner. The observed changes in mean and baseline plasma GH concentrations associated with 50 mg/kg BW CSH are consistent with somatostatin depletion; however, higher doses of CSH appear to disrupt GH secretion by an alternative mechanism.

Effects of diet forage:concentrate ratio and metabolizable energy intake on isolated rumen epithelial cell metabolism in vitro.

Crossbred wether lambs were used to assess the effect of altered forage:concentrate ratio and metabolizable energy intake on metabolism of substrates by ruminal epithelium using an isolated cell system. Lambs (n = 28; 20.1 +/- 3 kg BW) were assigned randomly to a factorial arrangement of dietary treatments consisting of either 75% forage or 75% concentrate fed once daily at either .099 or .181 Mcal ME x(kg BW(.75))(-1) x d(-1) for 52 d. After a 52-d feeding period, isolated rumen epithelial cells (IREC) were incubated in the presence of an oxidizable substrate with a single 14C label (acetate, propionate, butyrate, glucose, glutamate, and glutamine) at concentrations ranging from .1 to 50 mM, and substrate oxidation to 14CO2 or metabolism to beta-hydroxybutyrate (beta-HBA), acetoacetate, pyruvate, and lactate was determined. For all substrates, oxidation to CO2 was concentration-dependent and saturable within the physiological range. Differences in substrate oxidation to CO2 by IREC at specific substrate concentrations did not affect Vmax (maximal rate of substrate oxidation, nmol oxidized to CO2 x 1 x 10(6) cells(-1) x 90 min(-1)) and K(ox) (concentration of substrate at which half Vmax oxidation rate is achieved, mmoles/L) estimates for the dietary treatments. Production of beta-HBA from butyrate by IREC from the lambs fed 75% forage was not affected by ME intake; however, production was elevated by high ME intake of the 75% concentrate diet (diet x intake interaction; P < .02). Acetoacetate production from butyrate by IREC from lambs fed at high ME intake was greater (P = .001) than from those fed at low ME intake. Lactate and pyruvate production from glucose, glutamate, and propionate were generally unaffected by dietary treatment; however, rate of glutamine metabolism to lactate and pyruvate by IREC was increased with increased ME intake. The observed changes in metabolite production rates across groups did not affect the predicted Vmax and K(ox) parameter estimates. The estimated K(ox) values corroborate that VFA are the primary oxidizable fuels used by ruminal epithelial cells while illustrating that other substrates such as glucose, glutamate, and glutamine would not be expected to be oxidized extensively in vivo due to the high K(ox) relative to substrate concentrations in vivo. In conclusion, the capacity of isolated ruminal epithelial cells to oxidize substrates was largely unaffected by ME intake or dietary forage:concentrate ratio of the diet.

Effects of diet forage:concentrate ratio and metabolizable energy intake on visceral organ growth and in vitro oxidative capacity of gut tissues in sheep.

We used 28 crossbred wether lambs to determine the effects of dietary forage:concentrate ratio and metabolizable energy intake on visceral organ growth and oxidative capacity of gut tissues in lambs. Lambs were assigned randomly to a factorial arrangement of dietary treatments consisting of pelleted diets containing either 75% orchardgrass or 75% concentrate fed once daily at either .099 or .181 Mcal ME x (kg BW(.75))(-1) x d(-1). After a 52-d feeding period, lambs were slaughtered to obtain measurements of visceral organ mass and composition and oxidative capacity of isolated epithelial cells. Lamb performance, as measured by DMI, ADG, and efficiency of gain, was greater (P = .0001) for both diets at high ME intake. Likewise, lambs fed 75% concentrate gained faster and more (P < or = .01) efficiently than lambs fed 75% forage. Total digestive tract (TDT; includes rumen, reticulum, omasum, abomasum, and intestines) weight increased (P = .0001) with ME intake and was greater (P = .03) in lambs fed 75% forage than in those fed 75% concentrate. As a percentage of empty body weight (EBW), TDT weight increased with ME intake in lambs fed 75% forage, but it was unaffected by ME intake in lambs fed 75% concentrate (diet x intake, P = .03). Liver weight increased (P = .0001) with ME intake and was greater (P = .005) in lambs fed 75% concentrate vs 75% forage; however, liver weight as a percentage of EBW was increased (P = .0002) with ME intake but was unaffected by diet. Greater ME intake increased (P < or = .02) small intestinal (SI) epithelial and muscle mass of 15-cm sections, whereas jejunal epithelial mass was greater (P = .01) for lambs fed 75% forage vs 75% concentrate. Rumen epithelial concentrations of DNA and RNA increased (P < or = .02) with greater ME intake, whereas SI concentrations of DNA and RNA were largely unaffected by diet or ME intake. The activity of Na(+)-K(+)-ATPase increased in ileal epithelium (P < or = .02) with ME intake and concentrate in the diet, but activity in ruminal epithelium increased (P = .05) with concentrate. Total oxygen consumption by isolated ruminal and intestinal epithelial cells was unaffected by treatment. These data suggest that ME intake and level of dietary forage affect ruminal and intestinal growth via changes in cellular hyperplasia. Additionally, this study supports the concept that ME intake and diet composition alter gut energy expenditure, at least in part, through changes in mass rather than mass specific metabolism.

Cysteamine-induced depletion of somatostatin in sheep: time course of depletion and changes in plasma metabolites, insulin, and growth hormone.

Eight crossbred wethers (51 +/- 2 kg BW), surgically fitted with abomasal cannulas, were used to determine the extent and time course of cysteamine (CSH)-induced depletion of somatostatin (SRIF) in abomasal tissue and associated changes in plasma metabolites, insulin, and growth hormone (GH). Cysteamine was administered as a single i.v. bolus (50 mg.kg BW-1 x 10 min-1) on d 0. Abomasal biopsies were obtained on d -7, -3, 0, 1, 3, and 10. On d 0, additional biopsies were taken at 2, 4, and 8 h after CSH administration. Jugular blood samples were collected over 8 h at 15-min intervals on d -2, 0, and 1. Cysteamine administration decreased (P < .05) tissue SRIF on d 0 (2, 4, and 8 h), 1, and 3; maximal depletion (42 to 55% of Pre-treatment; Pre-trt) occurred during the initial 24 h, returning to Pre-trt by d 10. Gel chromatography of pooled -7 d abomasal tissue extracts showed five peaks of SRIF immunoreactivity; the predominate peak eluted in the same position as synthetic SRIF-14. Plasma glucose, lactate, and NEFA concentrations increased (P = .001) after CSH administration and reached peak at 2 h after treatment and declined to Pre-trt concentrations by 24 h. Insulin increased (P = .001) to a maximum at h 4 and returned to Pre-trt by 24 h. Mean and baseline GH were higher (P < .07) on day of CSH administration, and pulse amplitude was lower (P < .10) on d 0 and 1. These data show that CSH rapidly reduces SRIF in abomasal tissue in a reversible manner; suggesting that CSH-treated sheep may provide a SRIF-deficient model for studying the physiological role of SRIF in ruminants.

Adaptation to small intestinal starch assimilation and glucose transport in ruminants.

Four crossbred steers (380 +/- 6 kg) and seven Polypay wethers (40.3 +/- 6 kg) fitted with hepatic venous, hepatic portal, mesenteric venous and arterial, ruminal, and abomasal (steers) or duodenal (wethers) catheters were used in two crossover design experiments to evaluate adaptation to small intestinal starch and glucose transport. Steers were fed 8.6 kg/d and sheep were fed .9 kg/d of alfalfa hay in 12 equal portions and infused with an alpha-amylase partial starch hydrolysate (SH) either postruminally (adapted) or ruminally (unadapted) for 4 (steers; 40 g/h) or 5 (sheep; 6 g/h) days before measuring splanchnic flux of metabolites. On the day of flux measurements, ruminal SH infusion was switched to the postruminal site in unadapted animals. Flux measurements were made 3 to 6 h after switching infusion site for steers and 2 to 5 h for sheep. Phlorizin, a competitive inhibitor of Na/glucose cotransport, was then postruminally infused (550 and 500 mumol/h for steers and sheep, respectively) and flux measurements repeated from h 9 to 12 (steers) and h 7 to 10 (sheep). In the steers, adaptation increased (P < or = .09) portal-drained visceral (PDV) glucose release 26 mmol/h and decreased hepatic uptake of lactate 20 mmol/h. Abomasal infusion of phlorizin decreased (P = .01) net PDV glucose flux 40 mmol/h and concomitantly increased (P = .05) hepatic release of glucose by 47 mmol/h. In sheep, duodenal infusion of phlorizin increased (P < or = .005) portal and hepatic blood flow and decreased (P < or = .02) PDV release of glucose and lactate by 9 and .4 mmol/h, respectively. The liver released 4.2 mmol/h more (P = .09) glucose and removed 3.2 mmol/h more lactate (P = .09) glucose and removed 3.2 mmol/h more lactate (P = .001); arterial glucose decreased (P = .003) .75 mM in response to phlorizin. Phlorizin also caused loss of glucose in the urine of sheep (.09 mmol/h). Adaptation did not alter net splanchnic flux of glucose, lactate, oxygen, or alpha-amino N. These studies indicate that ruminants maintain the Na/glucose cotransporter when consuming little preformed alpha-glucosidic polymers and that the liver increases glucose release to compensate for reduced PDV delivery of glucose to maintain glucose homeostasis. In addition, the steers maintain at least 960 mmol/d and the sheep maintain at least 216 mmol/d of glucose transport capability regardless of adaptation.(ABSTRACT TRUNCATED AT 400 WORDS)

Effects of exogenous somatostatin and cysteamine on net nutrient flux across the portal-drained viscera and liver of sheep during intraduodenal infusion of starch hydrolysate and casein.

We used eight Polypay wethers (36 +/- .6 kg BW) fitted with hepatic portal, hepatic venous, mesenteric arterial and venous, and duodenal catheters in a crossover design experiment to determine the influence of somatostatin (SRIF) on splanchnic metabolism. Each crossover period consisted of 14 d, with net flux of nutrients and hormones (venoarterial differences x blood flow) measured on d 14. Before flux measurements, wethers received an i.v. dose (0 h) of either 0 (vehicle) or 50 mg x kg BW(-1) x 10 min(-1) cysteamine (CSH, SRIF-depleting agent) followed by a continuous duodenal infusion (h 10 to 22) of a starch hydrolysate-casein solution. Six sets of arterial, portal, and hepatic blood samples were obtained (h 12 to 16), after which a primed (10 microg), continuous jugular infusion of SRIF-14 (5.0 microg x kg BW(-1) x h(-1)) was initiated and sampling protocol repeated (h 18 to 22). Cysteamine administration increased (P < .01, vs control) portal and hepatic blood flow in the absence of exogenous SRIF (CSH x SRIF, P < .01). Net portal-drained viscera (PDV) release of glucose, alpha-amino N, ammonia N, beta-hydroxybutyrate, and oxygen consumption were decreased (P < or = .10) and lactate release increased (P = .005) during SRIF infusion. The CSH increased (P < .05) PDV release of beta-hydroxybutyrate and insulin and increased (P = .09, CSH alone vs control) net release of glucose in the absence of exogenous SRIF. Exogenous SRIF increased (P = .10) and CSH decreased (P = .09) net hepatic glucose output, whereas liver oxygen consumption was decreased (P = .04) with exogenous SRIF and increased (P = .01) with CSH. Net total splanchnic alpha-amino N release and oxygen consumption were decreased (P < .10) with exogenous SRIF, but CSH increased (P < .05) insulin release and oxygen consumption. These data provide initial evidence for a regulatory involvement of SRIF in visceral metabolism in ruminants.

Effects of ruminal and postruminal infusion of starch hydrolysate or glucose on the microbial ecology of the gastrointestinal tract in growing steers.

Forty crossbred steers were used to determine the effects of carbohydrate supply site on the indigenous bacteria of the gastrointestinal tract. Steers were fitted with ruminal and abomasal infusion catheters and assigned randomly to one of eight groups in a complete randomized block design. The experimental period was 36 d. Treatments included: 1) a pelleted basal diet fed at 0.163 Mcal ME x (kg BW(0.75)) x 1 x d(-1) (LE); 2) the basal diet fed at 0.215 Mcal ME x (kg BW(0.75)) (-1) x d(-1) (HE); 3) the basal diet fed at 0.163 Mcal ME x (kg BW(0.75))(-1) x d(-1) with ruminal infusion of starch hydrolysate (SH) (RSH); 4) the basal diet fed at 0.163 Mcal ME x (kg BW(0.75))(-1) x d(-1) with abomasal infusion of SH (ASH); and 5) the basal diet fed at 0.163 Mcal ME x (kg BW(0.75))(-1) x d(-1) with abomasal infusion of glucose (AG). The total volume ofinfusate (5 kg x site(-1) x d(-1)) was equalized across treatments and infusion sites by infusion of water. Glucose and SH were infused at rates of 14.35 and 12.64 g x (kg BW(0.75)) x d(-1), respectively. Ruminal, cecal, and fecal samples were obtained on d 36. Ruminal pH was low (5.79) in LE steers and unaffected (P > 0.10) by increased energy intake or carbohydrate infusion. Cecal and fecal pH were 6.93 and 7.00, respectively, for LE steers. Increasing energy intake (P < 0.10) and the rate of carbohydrate infusion (P < 0.01) significantly decreased cecal and fecal pH compared with LE. Ruminal counts of anaerobic bacteria in LE steers were 8.99 log10 cells/g and abomasal carbohydrate infusion had no affect (P > 0.10) on these numbers. However, ASH and AG steers had approximately 1.5 log10 cells/g more (P < 0.01) cecal and fecal anaerobic populations. Ruminal, cecal, and fecal aerobic bacterial counts were 40, 22, and 23%, respectively, lower than anaerobic counts. Generally, aerobic counts responded similarly to the anaerobic counts. Less than 1% of the anaerobic bacteria enumerated in the rumen, cecum, and feces were coliforms, and 97% of the coliforms were Escherichia coli. Carbohydrate infusions resulted in only numerical increases in fecal coliform and E. coli concentrations (P > 0.10). Fecal E. coli were highly acid sensitive in all steers, with less than 1% surviving a 1-h exposure to low pH (2.0). This suggests that cecal or fecal pH is not a good indicator of acid resistance, and it supports the concept that there are other factors that may induce acid resistance.

Influence of chlortetracycline and dietary protein level on visceral organ mass of growing beef steers.

Thirty-two beef steers (285 +/- 3 kg BW) were used to determine the effects of chlortetracycline and dietary protein level on visceral tissue mass, chemical composition, intestinal morphology, and proliferation rate indices. Steers were allotted randomly by weight to a factorial arrangement of dietary treatments consisting of either 10 or 13% CP diets top-dressed with a corn meal carrier (500 g/d) containing either 0 or 350 mg of chlortetracycline. After 84 d, steers were slaughtered and visceral organs removed and separated. Rinsed wet tissue mass was recorded; total RNA, total DNA, tissue DM, and tissue N content were determined; and tissue sections were prepared for immunohistochemical analysis. Thin tissue sections were evaluated to determine crypt depth and villus height as well as proliferation rate by immunohistochemical detection of the nuclear antigen Ki67. Rumen and abomasum weights and small intestinal length were greater (P < 0.04) in steers fed the 13% CP diet than in those fed the 10% CP diet on both an absolute weight basis and a percentage of empty BW. Chemical composition of the small intestinal and ruminal segments were largely unaffected by increased dietary protein. Increasing the dietary CP also increased the villus height in duodenal (P = 0.02) and the crypt depth of jejunal (P = 0.03) sections. Dietary administration of chlortetracycline decreased (P < 0.01) small intestinal weight both on absolute and empty BW bases. Nitrogen and RNA concentrations of the small intestinal segments were unaffected (P > 0.1) by dietary administration of subtherapeutic levels of chlortetracycline; however, because of increases (P < 0.05), or tendencies for an increase (P < 0.1), in the tissue content of DNA, the ratio of N to DNA was decreased (P < 0.05) or tended to be decreased (P < 0.1) in the small intestinal segments of the chlortetracycline-treated animals. The observed decrease in small intestinal epithelial mass does not appear to be due to alterations in cell proliferation rate but rather cell size. Consistent with this finding, cell proliferation, as determined by Ki67 antigen staining, was not affected by dietary treatment. Chlortetracycline administration decreased small intestinal mass that may be a result of decreased cell size.

Influence of abomasal carbohydrates on small intestinal sodium-dependent glucose cotransporter activity and abundance in steers.

Most animals adapt readily to increased supplies of carbohydrate in the intestinal lumen by increasing enzymes for degradation and increasing glucose transporter activity. However, the extent of upregulation of Na+-dependent glucose cotransporter 1 (SGLT1) activity and content in response to increased delivery of carbohydrate to the small intestinal lumen of ruminants is unclear. Therefore, an experiment was conducted to determine the effect of glucose and starch hydrolysate on the activity and abundance of SGLT1 in the small intestine of steers. In a randomized complete block design, 40 crossbred beef steers (243+/-2 kg BW) were fed 0.163 Mcal of ME/(kg BW0.75(d; W), 0.215 Mcal of ME/(kg BW0.75 x d; 2M), or 0.163 Mcal ME/(kg BW0.75 x d) and infused for 35 d into the rumen (R) or abomasum (A) with 12.6 g/(kg BW0.75 x d) of starch hydrolysate (S) or into the abomasum with 14.4 g/(kg BW0.75 x d) of glucose (G). Steers were slaughtered, and brush-border membrane vesicles were prepared from the small intestinal samples obtained from five equidistant sites along the intestine. Maltase activity in vesicles and homogenates differed with intestinal sampling site (quadratic, P < 0.001). Steers on the AG treatment yielded a greater intestinal maltase activity (38 nmol glucose x mg protein(-1) x min(-1)) compared with the AS, RS, W, or 2M treatments (34, 26, 23, and 23 nmol glucose x mg protein(-1) x min(-1) respectively [SEM = 3; P = 0.02]). Sodium-dependent glucose uptake averaged 18.4+/-3.94 pmol glucose/(mg protein x s) and was not affected by treatment, but uptake decreased distally along the intestine (P < 0.001). There was no effect of treatment on SGLT1 protein abundance, but SGLT1 protein abundance increased linearly from the duodenum to the ileum (P = 0.05). The inverse relationship between glucose uptake and SGLT1 abundance suggests that the regulation of brush border Na+-dependent glucose transport capacity is complex, involving factors other than the presence of luminal carbohydrate.

Portal and hepatic fluxes in sheep and concentrations in cattle ruminal fluid of 3-(4-hydroxyphenyl)propionic, benzoic, 3-phenylpropionic, and trans-cinnamic acids.

Extraction methods and HPLC procedures were developed for analysis of potential ruminal metabolites of dietary phenolics (reduced phenolics). Hepatic portal venous blood from wethers fed bromegrass, bermudagrass, ryegrass-wheat, and alfalfa hays also was analyzed for hippuric (HA), 3-(4-hydroxyphenyl)propionic (4OHPPA), benzoic (BA), 3-phenylpropionic (PPA), and t-cinnamic (CA) acids. Additionally, mesenteric arterial and hepatic venous blood was analyzed and, in conjunction with blood flow measurements, fluxes for portal-drained viscera (PDV) and liver were calculated. Ruminal fluid from four steers fed two levels of forage and two forage particle sizes in a Latin square design was analyzed for PPA and CA. 3-Phenylpropionic and benzoic acids were the most concentrated reduced phenolics identified in hepatic portal venous blood. Concentrations of PPA in ruminal fluid varied with ruminal disappearance of p-coumaric and ferulic acids. Additionally, hepatic portal venous concentrations of PPA were correlated (P < .05) with p-coumaric acid (r = .57) and ferulic acid (r = .67) intakes. Net release of PPA from PDV was observed, suggesting absorption of PPA from the gut. The liver removed PPA and BA with less efficiency. Given the relatively high concentrations of PPA in blood of ruminants, specific effects of this reduced phenolic on liver metabolism of ruminants should be assessed.