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Previous research has demonstrated that decreasing starch availability of steam-flaked corn by increasing flake density or increasing the degree of starch retrogradation influences in vitro gas production kinetics. However, it is unclear if increasing flake density or the degree of starch retrogradation influences end-products of in vitro ruminal fermentation (methane, volatile fatty acids, ammonia). The objective of this experiment was to evaluate the effects of increasing flake density and the degree of starch retrogradation on in vitro gas production kinetics, digestibility, and ruminal fermentation characteristics of steam-flaked corn. Three ruminally-cannulated steers were fed a high-concentrate diet and sampled for ruminal digesta for an in vitro fermentation experiment with a 5×2 factorial arrangement of treatments. Steam-flaked corn was produced to flake densities of 257, 296, 335, 373, and 412 g/L by adjusting the rolls of a steam-flaker. Samples were stored for 3 d at either 23°C to produce fresh steam-flaked corn or at 55°C in heat-sealed foil bags to produce retrograded steam-flaked corn. In vitro fermentation vessels were incubated for 24 h and then assessed for fermentation parameters including dry matter digestibility, volatile fatty acid concentrations, and total gas and methane production. Increasing the degree of starch retrogradation decreased (P < 0.01) the rate of gas production across all flake densities of steam-flaked corn but did not decrease the extent of gas production. In vitro methane production, dry matter digestibility, and microbial biomass concentration were not influenced by increasing flake density or starch retrogradation. Increasing the degree of starch retrogradation decreased (P = 0.03) the molar propionate proportion and increased (P < 0.06) the molar proportions of butyrate, isobutyrate, and isovalerate and the acetate:propionate ratio. Enzymatic starch availability of steam-flaked corn was positively correlated with mean propionate proportion (r2 = 0.93) and negatively correlated with the mean butyrate proportion (r2 = 0.89). Results from the current study demonstrate that increasing the degree of starch retrogradation of steam-flaked corn decreased the rate of in vitro gas production and altered volatile fatty acid profiles in the ruminal fermentation media., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Fescue toxicosis is a syndrome occurring from the consumption of endophyte-infected tall fescue and results in substantial economic losses to the beef industry primarily from reduced growth accompanied by decreased dry matter intake (DMI); however, the associations characterizing this reduction in DMI have yet to be elucidated. The objective of this experiment was to identify endocrine changes associated with intake regulation post-consumption of endophyte-infected tall fescue seed (E+). Twelve Holstein steers were stratified by body weight and assigned to 1 of 3 treatments (n=4): 0 ppm ergovaline (ERV), 1.8 ppm ERV, or 2.7 ppm ERV. Treatments were achieved by combining differing proportions of ground E+ and non-endophyte-infected tall fescue seed. Steers were adapted to their diets for 7 d followed by a 7 d DMI collection period. Within treatment, steers were assigned to a sampling day (d 16 or d 17). Blood samples were collected every 20 min for 8 h, beginning 1 h before feeding. Intake data was analyzed using the MIXED procedure of SAS 9.4 (SAS Inst. Inc., Cary, NC) with treatment, day, and the interaction as fixed effects. Hormone and metabolite data were analyzed with the fixed effect of treatment, time, and the interaction including time as a repeated measure and orthogonal contrasts. Dry matter intake was linearly decreased with increasing ERV in the diet (P < 0.001). Insulin and leptin concentrations exhibited a quadratic effect (P = 0.018 and P = 0.005) with insulin concentrations highest for the 2.7 ppm treatment and leptin concentrations highest for the 1.8 ppm treatment. No differences were detected for active ghrelin or β-hydroxybuytrate concentrations among treatment groups. Further, steers consuming both the 1.8 and 2.7 ppm ERV treatments had lower prolactin concentrations compared to the 0 ppm treatment (quadratic, P= 0.019). Glucose concentrations had a tendency for a linear increase as ERV concentrations increased (P = 0.091). A treatment × time interaction (P = 0.002) was noted in NEFA concentrations, with the 1.8 ppm ERV treatment showing increased pre-feeding concentrations, and the 2.7 ppm ERV treatment exhibiting elevated NEFA concentrations as time post-feeding progressed. The results suggest that E+ consumption reduces intake likely through alterations in intake-related hormones and post-absorptive metabolism and contributes to our current understanding of E+ effects on intake reduction while providing avenues for future research., (Copyright © 2024 Elsevier Inc. All rights reserved.)

We previously demonstrated that postruminal casein infusion and exogenous glucagon-like peptide 2 (GLP-2) administration independently stimulated growth and carbohydrase activity of the pancreas and jejunal mucosa in cattle. The objective of the current study was to profile the jejunal mucosal transcriptome of cattle using next-generation RNA sequencing in response to postruminal casein infusion and exogenous GLP-2. Twenty-four Holstein steers [250 ± 23.1 kg body weight (BW)] received a continuous abomasal infusion of 3.94 g raw corn starch/kg of BW combined with either 0 or 1.30 g casein/kg of BW for 7 d. Steers received subcutaneous injections at 0800 and 2000 h to provide either 0 or 100 μg GLP-2/kg of BW per day. At the end of the 7-d treatment period, steers were slaughtered for collection of the jejunal mucosa. Total RNA was extracted from jejunal mucosal tissue, strand-specific cDNA libraries were prepared, and RNA sequencing was conducted to generate 150-bp paired-end reads at a depth of 40 M reads per sample. Differentially expressed genes (DEG), KEGG pathway enrichment, and gene ontology enrichment were determined based on the FDR-corrected P-value (padj). Exogenous GLP-2 administration upregulated (padj < 0.05) 667 genes and downregulated 1,101 genes of the jejunal mucosa. Sphingolipid metabolism, bile secretion, adherens junction, and galactose metabolism were among the top KEGG pathways enriched with upregulated DEG (padj < 0.05) in response to exogenous GLP-2 administration. The top gene ontologies enriched with upregulated DEG (padj < 0.05) in response to exogenous GLP-2 administration included nutrient metabolic processes, brush border and bicellular tight junction assembly, and enzyme and transporter activities. Exogenous GLP-2 administration increased or tended to increase (padj < 0.10) brush border carbohydrase (MGAM, LCT, TREH), hexose transporter (SLC5A1, SLC2A2), and associated transcription factor (HNF1, GATA4, KAT2B) mRNA expression of the jejunal mucosa. Gene ontologies and KEGG pathways that were downregulated (padj < 0.05) in response to exogenous GLP-2 were related to genetic information processing. Postruminal casein infusion downregulated (padj < 0.05) 7 jejunal mucosal genes that collectively did not result in enriched KEGG pathways or gene ontologies. This study highlights some of the transcriptional mechanisms associated with increased growth, starch assimilation capacity, and barrier function of the jejunal mucosa in response to exogenous GLP-2 administration., Competing Interests: The authors have declared that no competing interests exist., (Copyright: This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.)

To better understand mechanisms of serotonin- (5-HT) mediated vasorelaxation, isolated lateral saphenous veins from cattle were assessed for vasoactivity using myography in response to increasing concentrations of 5-HT or selective 5-HT receptor agonists. Vessels were pre-contracted with 1 × 10 -4  M phenylephrine and exposed to increasing concentrations of 5-HT or 5-HT receptor agonists that were selective for 5-HT 1B , 5-HT 2B , 5-HT 4 , and 5-HT 7 . Vasoactive response data were normalized as a percentage of the maximum contractile response induced by the phenylephrine pre-contraction. At 1 × 10 -7  M 5-HT, a relaxation was observed with an 88.7% decrease (p < 0.01) from the phenylephrine maximum. At 1 × 10 -4  M 5-HT, a contraction was observed with a 165% increase (p < 0.01) from the phenylephrine maximum. Increasing concentrations of agonists selective for 5-HT 2B , 5-HT 4 , or 5-HT 7 resulted in a 27%, 92%, or 44% (p < 0.01) decrease from the phenylephrine maximum, respectively. Of these 5-HT receptor agonists, the selective 5-HT 4 receptor agonist resulted in the greatest potency (-log EC 50 ) value (6.30) compared with 5-HT 2B and 5-HT 7 receptor agonists (4.21 and 4.66, respectively). To confirm the involvement of 5-HT 4 in 5-HT-mediated vasorelaxation, blood vessels were exposed to either DMSO (solvent control) or a selective 5-HT 4 antagonist (1 × 10 -5  M) for 5-min prior to the phenylephrine pre-contraction and 5-HT additions. Antagonism of the 5-HT 4 receptor attenuated the vasorelaxation caused by 5-HT. Approximately 94% of the vasorelaxation occurring in response to 5-HT could be accounted for through 5-HT 4 , providing strong evidence that 5-HT-mediated vasorelaxation occurs through 5-HT 4 activation in bovine peripheral vasculature., (© 2024 The Author(s). Physiological Reports published by Wiley Periodicals LLC on behalf of The Physiological Society and the American Physiological Society. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.)

The present study evaluated the effects of sodium butyrate (SB) supplementation on exocrine and endocrine pancreatic development in dairy calves. Fourteen male Holstein calves were alimented with either milk or milk supplemented with SB for 70 days. Pancreases were collected for analysis including staining, immunofluorescence, electron microscopy, qRT-PCR, Western blotting, and proteomics. Results indicated increased development in the SB group with increases in organ size, protein levels, and cell growth. There were also exocrine enhancements manifested as higher enzyme activities and gene expressions along with larger zymogen granules. Endocrine benefits included elevated gene expression, more insulin secretion, and larger islets, indicating a rise in β-cell proliferation. Proteomics and pathway analyses pinpointed the G protein subunit alpha-15 as a pivotal factor in pancreatic and insulin secretion pathways. Overall, SB supplementation enhances pancreatic development by promoting its exocrine and endocrine functions through G protein regulation in dairy calves.

The impact of ergot toxicosis on livestock industries is detrimental and treatments are needed in many countries. The objective of this study was to evaluate the effects of acute exposure to ergot alkaloids and 5-hydroxytryptophan (5-HTP) supplementation on feed intake, serotonin metabolism, and blood metabolites in cattle. Eight Holstein steers (538 ± 18 kg) fitted with ruminal cannulas were used in a replicated 4 × 4 Latin Square design experiment with a 2 × 2 factorial treatment structure. The treatments were the combination of 0 (E-) or 15 µg ergovaline/kg BW (E+) and 0 (5HTP-) or 0.5 mg of 5-hydroxy-l-tryptophan/kg BW (5HTP+) administered daily for 6 d. Toxic endophyte-infected tall fescue seed was used to supply the daily dose of ergovaline. Endophyte-free seed was used to equalize seed intake between treatments. Ground seed was placed into the rumen immediately before feeding. The 5-HTP was dissolved in water and infused into the abomasum via the reticulo-omasal orifice. Blood was collected from a jugular vein catheter at 0, 1, 2, 4, 8, and 24 h after treatment administration. Ergovaline without 5-HTP (E+/5HTP-) decreased dry matter intake (DMI) in comparison to steers without ergovaline and 5-HTP (E-/5HTP-). However, 5-HTP infusion in association with ergovaline (E+/5HTP+) normalized the DMI. Although E + did not affect (P > 0.05) the area under the curve (AUC) of serum 5-HTP, 5-hydroxyindoleacetic acid, tryptophan, and kynurenine, serum and plasma serotonin concentrations were decreased (P < 0.05). The infusion of 5-HTP increased (P < 0.05) the AUC of serum 5-HTP, serum and plasma serotonin, and serum 5-hydroxyindoleacetic acid. In conclusion, acute exposure to ergot alkaloids reduced DMI and circulating serotonin in cattle but 5-HTP administration showed potential to normalize both circulating serotonin and feed intake., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Consumption of ergot alkaloids from endophyte-infected tall fescue results in losses to the livestock industry in many countries and a means to mitigate these losses is needed. The objective of this study was to evaluate intra-abomasal infusion of the dopamine precursor, levodopa (L-DOPA), on dopamine metabolism, feed intake, and serum metabolites of steers exposed to ergot alkaloids. Twelve Holstein steers (344.9 ± 9.48 kg) fitted with ruminal cannula were housed with a cycle of heat challenge during the daytime (32 °C) and thermoneutral at night (25 °C). The steers received a basal diet of alfalfa cubes containing equal amounts of tall fescue seed composed of a mixture of endophyte-free (E-) or endophyte-infected tall fescue seeds (E+) equivalent to 15 µg ergovaline/kg body weight (BW) for 9 d followed by intra-abomasal infusion of water (L-DOPA-) or levodopa (L-DOPA+; 2 mg/kg BW) for an additional 9 d. Afterward, the steers were pair-fed for 5 d to conduct a glucose tolerance test. The E+ treatment decreased (P = 0.005) prolactin by approximately 50%. However, prolactin increased (P = 0.050) with L-DOPA+. Steers receiving E+ decreased (P < 0.001) dry matter intake (DMI); however, when supplemented with L-DOPA+ the decrease in DMI was less severe (L-DOPA × E, P = 0.003). Also, L-DOPA+ infusion increased eating duration (L-DOPA × E, P = 0.012) when steers were receiving E+. The number of meals, meal duration, and intake rate were not affected (P > 0.05) by E+ or L-DOPA+. The L-DOPA+ infusion increased (P < 0.05) free L-DOPA, free dopamine, total L-DOPA, and total dopamine. Conversely, free epinephrine and free norepinephrine decreased (P < 0.05) with L-DOPA+. Total epinephrine and total norepinephrine were not affected (P > 0.05) by L-DOPA+. Ergot alkaloids did not affect (P > 0.05) circulating free or total L-DOPA, dopamine, or epinephrine. However, free and total norepinephrine decreased (P = 0.046) with E+. Glucose clearance rates at 15 to 30 min after glucose infusion increased with L-DOPA+ (P < 0.001), but not with E+ (P = 0.280). Administration of L-DOPA as an agonist therapy to treat fescue toxicosis provided a moderate increase in DMI and eating time and increased plasma glucose clearance for cattle dosed with E+ seed., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Increasing luminal carbohydrate flow decreases pancreatic α-amylase activity but can increase jejunal maltase activity, suggesting that regulation of carbohydrase activity is perhaps uncoordinated in response to luminal carbohydrate flow. Increasing luminal casein flow increases pancreatic α-amylase activity in cattle, and exogenous glucagon-like peptide 2 (GLP-2) has been shown to increase small intestinal α-glucosidase activity in nonruminants., Objectives: The objective was to evaluate the effects of postruminal casein infusion, exogenous GLP-2, or their combination on endogenous pancreatic and small intestinal carbohydrase activity in cattle postruminally infused with starch., Methods: Holstein steers [n = 24; 250 ± 23 kg body weight (BW)] received a continuous abomasal infusion of 3.94 g raw corn starch/kg of BW combined with either 0 or 1.30 g casein/kg of BW. Steers received subcutaneous injections in 2 equal portions daily of excipient (0.5% bovine serum albumin) or 100 μg GLP-2/kg of BW per day. At the end of the 7-d treatment period, steers were slaughtered for tissue collection. Data were analyzed using the MIXED procedure of SAS version 9.4 (SAS Institute Inc.)., Results: Postruminal casein infusion increased (P ≤ 0.03) pancreatic mass by 12.6%, total pancreatic α-amylase activity by 50%, and postruminal starch disappearance from 96.7% to 99.3%. Exogenous GLP-2 increased (P < 0.01) total small intestinal and mucosal mass by 1.2 kg and 896 g, respectively. Relative to control, GLP-2 and casein + GLP-2 increased (P = 0.04) total small intestinal α-glucosidase activity by 83.5%. Total small intestinal maltase, isomaltase, and glucoamylase activity was 90%, 100%, and 66.7% greater for GLP-2 and casein + GLP-2 steers compared with control., Conclusions: Casein increased pancreatic α-amylase activity, GLP-2 increased small intestinal α-glucosidase activity, and the combination of casein and GLP-2 increased both pancreatic α-amylase activity and small intestinal α-glucosidase activity. This novel approach provides an in vivo model to evaluate effects of increasing endogenous carbohydrase activity on small intestinal starch digestion., (Copyright © 2023 American Society for Nutrition. Published by Elsevier Inc. All rights reserved.)

Four experiments were conducted to evaluate sources of anti-coccidial compounds and phytogenic saponin extracts on in vitro and in vivo ruminal fermentation and CH 4 production at multiple inclusion levels. In experiment 1, eight steers were fed either a finishing diet or a finishing diet supplemented with 0.5 mg/kg BW decoquinate (DCQ) and 3.33 mg/kg BW Yucca schidigera extract (YSE), and respiratory gas exchange was measured. In experiment 2, four ruminally-cannulated steers were fed the same treatments as experiment 1, and ruminal fermentation was evaluated. Anti-coccidial sources (experiment 3; monensin, DCQ, amprolium) and saponin sources (experiment 4; YSE, Quillaja saponaria extract) and levels were evaluated for effects on in vitro ruminal fermentation and CH 4 production. DCQ + YSE supplementation did not influence ( p ≥ 0.24) in vivo respiratory gas consumption/production, in situ DM degradation, or liquid passage kinetics. Ruminal propionate proportion tended to increase ( p = 0.09) with DCQ + YSE. Monensin decreased ( p ≤ 0.04) in vitro acetate:propionate and CH 4 production; saponin supplementation linearly increased ( p < 0.01) propionate proportion but did not influence ( p ≥ 0.38) in vitro CH 4 production. Saponins and non-antibiotic anti-coccidials did not influence in vitro or in vivo CH 4 production with finishing diets.

Grain processing such as particle size, flake density, or starch retrogradation can influence ruminal degradability characteristics; however, it is unclear how exogenous α-amylase supplementation interacts with different processed grains. Four experiments were conducted to compare the effects of Aspergillus oryzae fermentation extract (Amaize; Alltech Biotechnology Inc., Nicholasville, KY) supplementation on in vitro gas production kinetics of grain substrates with different processing methods that are common in the feedlot industry. In experiment 1, corn processing (dry-rolled, high-moisture, steam-flaked) and Amaize supplementation (0 or 15 U α-amylase activity/100 mL) were evaluated in a 3 × 2 factorial arrangement of treatments. The rate of gas production for dry-rolled corn was higher (P < 0.001) with Amaize supplementation. In experiment 2, flake density (296, 322, 348, 373, and 399 g/L) and starch retrogradation (storage in heat-sealed foil bags for 3 d at 23 or 55°C) were evaluated in a 5 × 2 factorial arrangement of treatments. There was a flake density × starch retrogradation interaction (P < 0.01) for the rate of gas production because the decrease in the rate of gas production with starch retrogradation was greater at lighter flake densities compared with heavier flake densities. In experiment 3, Amaize supplementation was evaluated across flake densities of nonretrograded steam-flaked corn (stored at 23°C) used in experiment 2. There was a flake density × Amaize interaction (P < 0.01) for the rate of gas production where Amaize supplementation resulted in a lower rate of gas production at lighter flake densities (296, 322, and 348 g/L) but a higher rate of gas production at heavier flake densities (373 and 399 g/L). In experiment 4, Amaize supplementation was evaluated across flake densities of retrograded steam-flaked corn (stored at 55°C) used in experiment 2. Gas production was lower after 24 h with Amaize supplementation for retrograded flakes produced to a density of 322 and 399 g/L while Amaize supplementation did not influence gas production at 24 h at other flake densities. There was a flake density × Amaize interaction for the rate of gas production because Amaize supplementation resulted in a faster (P < 0.01) rate of gas production for all flake densities except retrograded flakes produced to a density of 296 g/L. Enzymatic starch availability was positively correlated with the rate of gas production. These data demonstrate that supplementation of 15 U/100 mL of Amaize resulted in greater rates of gas production for dry-rolled corn, corn steam-flaked to heavier densities, and retrograded steam-flaked corn., (© The Author(s) 2023. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Ergovaline (ERV), produced in toxic endophyte-infected tall fescue, causes potent vasoconstriction of bovine peripheral and visceral vasculature. Ergovaline acts as both an agonist and an antagonist in bovine gut blood vessels through serotonin (5-HT) receptors and it appears that the type of action could be influenced by the extent of ERV exposure. Because it was unclear how the duration of ERV exposure influences 5-HT-mediated vasoactivity, experiments were designed to evaluate how simultaneous or prior ERV exposure influenced 5-HT-mediated vasoactivity of mesenteric artery (MA) and vein (MV) segments from Holstein steers (N = 10). Vessels were incubated in Krebs-Henseleit buffer containing 0, 0.01, or 0.1 μM ERV for 24 h prior to the 5-HT dose-response or exposed to fixed concentrations of 0, 0.01, or 0.1 μM ERV simultaneously during the 5-HT dose-response. Vessels were suspended in chambers of a multimyograph containing Krebs-Henseleit buffer and equilibrated to 1 g tension for 90 min. Vessels were exposed to increasing concentrations of 5-HT (5 × 10-8 M to 1 × 10-4 M) every 15 min and contractile responses were normalized as a percentage of the maximum contractile response induced by 120 mM KCl reference addition. Two-way analysis of variance was used to separately analyze data for each vessel type and duration of exposure using the MIXED procedure of SAS. When 5-HT concentration increased from 5 × 10-8 to 1 × 10-6 M, simultaneous addition of 0.1 μM ERV increased (P < 0.01) the contractile response of MV compared with additions of 0 and 0.01 μM ERV. At 1 × 10-4 M 5-HT, the simultaneous presence of 0.01 and 0.1 μM ERV decreased (P < 0.01) the contractile response of both MA and MV compared with 0 μM ERV addition. As 5-HT concentrations increased, the contractile response increased (P < 0.01) in both MA and MV with no previous ERV exposure, but decreased in MA and MV with 24 h prior exposure to 0.01 and 0.1 μM ERV. These data demonstrate that the duration of ERV exposure influences 5-HT-mediated vasoconstriction and likely vasorelaxation in bovine mesenteric vasculature. If ERV and 5-HT exposure occur simultaneously, ERV can act as a partial agonist of 5-HT-mediated vasoconstriction. If 5-HT exposure occurs after blood vessels have had prior ERV exposure, it appears that 5-HT may induce vasorelaxation of blood vessels. More research is needed to identify cellular and molecular mechanisms involved with 5-HT-mediated vasoactivity., (Published by Oxford University Press on behalf of the American Society of Animal Science 2023.)

Five ruminally cannulated steers (body weight = 390 ± 7.86 kg) were used in three experiments to evaluate effects of corn processing, flake density, and starch retrogradation on in situ ruminal degradation. In experiment 1, corn was left whole or processed with no screen, ground through a 6-mm screen, or ground through a 1-mm screen. In experiment 2, we produced steam-flaked corn at four densities: 309, 335, 360, and 386 g/L. These four flake densities were sifted for 20 s through a 4-mm screen to produce two particle sizes within each flake density: sifted flakes (>4 mm) and sifted fines (<4 mm). In experiment 3, sifted flakes (335 g/L) were stored for 3-d at either 23 °C (starch availability = 55%) or 55 °C to induce starch retrogradation (starch availability = 41%). All samples for each of the three experiments were weighed into nylon bags and ruminally incubated for 0-h to estimate the soluble fraction. The residue remaining was analyzed for nutrient composition. In experiment 1, whole shelled corn had lesser (P < 0.01) ruminal solubility of all nutrients measured compared with ground corn. Corn ground with a screen (6 and 1 mm) had greater (P < 0.01) ruminal solubility of all nutrients measured compared with corn ground with no screen. Corn ground through a 1-mm screen had greater (P < 0.03) ruminal solubility of DM, total starch, CP, ADF, AHF, P, Mg, K, S, Zn, Fe, and Mn compared with corn ground through a 6-mm screen. In experiment 2, increasing flake density linearly decreased (P < 0.02) the soluble fraction of DM, total starch, CP, ADF, AHF, P, K, S, and Zn of sifted flakes. The soluble DM fraction of sifted fines tended to decrease (P = 0.06) linearly with increasing flake density. Total starch, CP, NDF, and Zn soluble fractions of sifted fines were not influenced by flake density. In experiment 3, storage of sifted flakes in heat-sealed foil bags at 55 °C for 3-d decreased (P < 0.04) the soluble fractions of DM, total starch, CP, NDF, P, Mg, K, S, and Fe. With each increase in the degree of corn processing, there was an increase in the solubility of nutrients. Increasing flake density can decrease ruminal solubility of flakes; however, the soluble fraction of sifted fines is not influenced as much by changes in flake density. Inducing starch retrogradation decreases ruminal solubility of starch, nonstarch OM, and minerals., (© The Author(s) 2022. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Due to the potential risk for cannabidiol (CBD) to negatively impact the immune system, the objective of the current study was to evaluate the effect of CBD on the canine immune response to immunization with a novel antigen, keyhole limpet hemocyanin (KLH). Thirty-two dogs (22.4 ± 6.3 kg BW) were utilized in a completely randomized design with treatments consisting of 5 mg CBD/kg BW/d and a control administered orally via treats. After a 7-d acclimation to treatments, dogs were immunized with 10 mg/dog of KLH via intramuscular injection into the semimembranosus muscle region, which was repeated in 14 d. Blood samples were collected at baseline and weekly for 28 d after initial KLH immunization for analysis of hematology, serum chemistry, and immunoglobulins. Data were analyzed using the MIXED procedure in SAS including the fixed effects of treatment, day, and the treatment by day interaction. Both primary and secondary KLH immunization produced robust immune responses. Most hematological and serum chemistry variables remained within normal reference ranges for dogs across both treatments throughout the study. Alkaline phosphatase, while within normal reference range and similar between treatments at baseline and on d 7 (P = 0.994 and 0.183, respectively), was elevated for CBD-treated dogs versus control on d 14, 21, and 28 (P = 0.006, 0.027, and 0.014, respectively). Both total and KLH-specific IgG and IgM were similar between treatments throughout the study (P > 0.05), although total IgM peaked earlier in control dogs compared to those receiving CBD. Despite the minor shift in the timing of the total IgM peak, CBD did not appear to exhibit humoral immunosuppressive effects when supplemented at 5 mg/kg BW/d. However, this work does highlight the potential for CBD to alter liver function and the need for further safety evaluations of CBD use in dogs utilizing longer-term studies and multiple CBD doses., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

Ergot alkaloid mycotoxins interfere in many functions associated with serotonergic neurotransmitters. Therefore, the objective was to evaluate whether the association of serotonin (5-hydroxytryptamine, 5-HT) and ergot alkaloids during a 24 h pre-incubation could affect the vascular contractile response to ergot alkaloids. To evaluate the effects of 24 h exposure to 5-HT and ergot alkaloids (ergovaline, ERV), two assays were conducted. The first assay determined the half-maximal inhibitory concentration (IC 50 ) following the 24 h pre-exposure period, while the second assay evaluated the effect of IC 50 concentrations of 5-HT and ERV either individually or in combination. There was an interaction between previous exposure to 5-HT and ERV. Previous exposure to 5-HT at the IC 50 concentration of 7.57 × 10 -7 M reduced the contractile response by more than 50% of control, while the exposure to ERV at IC 50 dose of 1.57 × 10 -10 M tended to decrease ( p = 0.081) vessel contractility with a response higher than 50% of control. The 24 h previous exposure to both 5-HT and ERV did not potentiate the inhibitory response of blood vessels in comparison with incubation with each compound alone. These results suggest receptor competition between 5-HT and ERV. More studies are necessary to determine the potential of 5-HT to treat toxicosis caused by ergot alkaloids.

Five ruminally cannulated steers (body weight = 390 ± 7.86 kg) were used in two experiments to evaluate the effects of flake density and starch retrogradation on in situ ruminal degradation of steam-flaked corn. In experiment 1, sifted flakes with flake densities of 257, 296, 335, 373, and 412 g/L (enzymatic starch availabilities: 87%, 76%, 66%, 43%, and 49%, respectively) were evaluated in a randomized complete block design experiment. In experiment 2, the experimental design was a randomized complete block design with a 3 × 2 factorial arrangement of treatments. Three steam-flaked corn fractions corresponding to different particle sizes were used: flakes + fines (not sifted; >4 and <4 mm), sifted flakes (>4 mm), and sifted fines (<4 mm). Particle size fractions were stored for 3 d at either 23 °C or 55 °C (starch availabilities averaged across particle sizes: 53.3% and 25.5%, respectively) in heat-sealed foil bags. Samples were ruminally incubated for 0, 3, 6, 12, 24, 48, 72, or 96 h. Degradation data were modeled to obtain the rate and extent of degradation and passage rate was set to 6% per hour. In experiment 1, the rate of degradation decreased linearly (P < 0.01) and in situ ruminal dry matter (DM) degradability decreased linearly (P < 0.01) from 78.9% to 57.3% as flake density increased from 257 to 412 g/L. In experiment 2, storage of steam-flaked corn samples at 55 °C for 3 d decreased (P < 0.01) the rate of degradation by 37.6% across all particle sizes. Storing samples at 55 °C for 3 d decreased (P < 0.01) in situ ruminal DM degradability of flakes + fines, sifted flakes, and sifted fines by 20.9%, 22.6%, and 14.7%, respectively. Using data from experiment 1 and 2, enzymatic starch availability of sifted flakes was positively correlated (R2 = 0.97; P < 0.01) with in situ ruminal DM degradability. The results demonstrate that decreased starch availability resulting from either starch retrogradation or increased flake density is associated with decreased ruminal digestibility. Decreases in starch availability and in situ ruminal degradability may indicate that increasing flake density or starch retrogradation could potentially alter the site of digestion in cattle. Using prediction equations, decreases in ruminal starch digestibility of steam-flaked corn caused by increasing flake density or increasing starch retrogradation could increase energetic efficiency, depending on the rate of passage and if small intestinal starch digestibility is maintained., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Subacute rumen acidosis (SARA) occurs when highly fermentable carbohydrates are introduced into the diet, decreasing pH and disturbing the microbial ecology of the rumen. Rumen amylolytic bacteria rapidly catabolize starch, fermentation acids accumulate in the rumen and reduce environmental pH. Historically, antibiotics (e.g., monensin, MON) have been used in the prevention and treatment of SARA. Biochanin A (BCA), an isoflavone produced by red clover (Trifolium pratense), mitigates changes associated with starch fermentation ex vivo. The objective of the study was to determine the effect of BCA on amylolytic bacteria and rumen pH during a SARA challenge. Twelve rumen fistulated steers were assigned to 1 of 4 treatments: HF CON (high fiber control), SARA CON, MON (200 mg d-1), or BCA (6 g d-1). The basal diet consisted of corn silage and dried distiller's grains ad libitum. The study consisted of a 2-wk adaptation, a 1-wk HF period, and an 8-d SARA challenge (d 1-4: 40% corn; d 5-8: 70% cracked corn). Samples for pH and enumeration were taken on the last day of each period (4 h). Amylolytic, cellulolytic, and amino acid/peptide-fermenting bacteria (APB) were enumerated. Enumeration data were normalized by log transformation and data were analyzed by repeated measures ANOVA using the MIXED procedure of SAS. The SARA challenge increased total amylolytics and APB, but decreased pH, cellulolytics, and in situ DMD of hay (P < 0.05). BCA treatment counteracted the pH, microbiological, and fermentative changes associated with SARA challenge (P < 0.05). Similar results were also observed with MON (P < 0.05). These results indicate that BCA may be an effective alternative to antibiotics for mitigating SARA in cattle production systems., Competing Interests: The authors have declared that no competing interests exist.

Despite the increased interest and widespread use of cannabidiol (CBD) in humans and companion animals, much remains to be learned about its effects on health and physiology. Metabolomics is a useful tool to evaluate changes in the health status of animals and to analyze metabolic alterations caused by diet, disease, or other factors. Thus, the purpose of this investigation was to evaluate the impact of CBD supplementation on the canine plasma metabolome. Sixteen dogs (18.2 ± 3.4 kg BW) were utilized in a completely randomized design with treatments consisting of control and 4.5 mg CBD/kg BW/d. After 21 d of treatment, blood was collected ~2 h after treat consumption. Plasma collected from samples was analyzed using CIL/LC-MS-based untargeted metabolomics to analyze amine/phenol- and carbonyl-containing metabolites. Metabolites that differed - fold change (FC) ≥ 1.2 or ≤ 0.83 and false discovery ratio (FDR) ≤ 0.05 - between the two treatments were identified using a volcano plot. Biomarker analysis based on receiver operating characteristic (ROC) curves was performed to identify biomarker candidates (area under ROC ≥ 0.90) of the effects of CBD supplementation. Volcano plot analysis revealed that 32 amine/phenol-containing metabolites and five carbonyl-containing metabolites were differentially altered (FC ≥ 1.2 or ≤ 0.83, FDR ≤ 0.05) by CBD; these metabolites are involved in the metabolism of amino acids, glucose, vitamins, nucleotides, and hydroxycinnamic acid derivatives. Biomarker analysis identified 24 amine/phenol-containing metabolites and 1 carbonyl-containing metabolite as candidate biomarkers of the effects of CBD (area under ROC ≥ 0.90; P < 0.01). Results of this study indicate that 3 weeks of 4.5 mg CBD/kg BW/d supplementation altered the canine metabolome. Additional work is warranted to investigate the physiological relevance of these changes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Morris, Kitts-Morgan, Spangler, Ogunade, McLeod and Harmon.)

Measuring enzymatic starch availability is commonly used as a quality control method to ensure steam-flaked corn manufacturing consistency in commercial cattle feeding operations. However, starch availability estimates can be variable. We conducted five experiments to evaluate factors influencing starch availability estimates of steam-flaked corn. In Exp. 1, sample handling methods were evaluated. Sifted flakes were immediately placed into a plastic bag, air equilibrated for 240 min, oven-dried, or freeze-dried. Directly oven-drying samples at 55°C decreased (P < 0.01) starch availability compared to other sample handling methods. In Exp. 2, sifted flakes were air equilibrated for 0, 15, 30, 60, 120, or 240 min. Air equilibration time did not influence (P ≥ 0.54) starch availability. In Exp. 3, samples were evaluated for effects of sifting through a 4-mm screen (flakes + fines vs. sifted flakes) and air equilibration time (0 vs. 240 min). Both sifting steam-flaked corn samples and air equilibration for 240 min increased starch availability (P < 0.01 and P = 0.02, respectively). In Exp. 4, we evaluated the effects of air equilibration time (0 vs. 240 min) on the two sifted portions (sifted flakes vs. sifted fines). There was an air equilibration time × sifted portion interaction for starch availability because air equilibration time increased (P < 0.01) starch availability of sifted fines but did not influence starch availability of sifted flakes. Concentrations of crude protein, soluble crude protein, neutral and acid detergent fiber, ether extract, and acid-hydrolyzed fat, Ca, P, K, Mg, S, Fe, Zn, Mg, and Cu were greater (P < 0.01) for sifted fines compared to sifted flakes. Starch availability and total starch concentration were greater (P < 0.01) for sifted flakes compared to sifted fines. In Exp. 5, effects of air equilibration time (0 vs. 240 min) and storage temperature (23°C vs. 55°ºC) on flakes + fines were evaluated. Storage of flakes + fines in heat-sealed foil bags at 55°C for 3-d decreased (P < 0.01) starch availability by 40.7%. Sifted flakes contained less moisture, greater total starch concentrations, and greater starch availability than sifted fines. Moisture, sifting, air equilibration time, and storage temperature influence starch availability of steam-flaked corn. Adoption of the strategies discussed in the current study will lead to more consistent estimates of starch availability., (© The Author(s) 2021. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Studies of serotonin in animal husbandry has received growing interest. However, there is limited information about serotonin manipulation using 5-HTP administered postruminally and its residual effects in cattle. The objective of this study was to evaluate the effectiveness of 5-HTP infused into the abomasum for enhancing circulating serotonin in cattle. Four Holstein steers (487 ± 7.6 kg) fitted with ruminal cannulas were used in a 4 × 4 Latin Square design experiment. The treatments were intra-abomasal infusion of 5-HTP at 0, 0.25, 0.5, and 1 mg/kg BW. Blood was collected from the jugular vein of each steer at -60, -30, 0, 30, 60, 120, 240, and 480 min from 5-HTP infusion for basal and short term evaluation and, at 1, 2, 4, and 7 d after 5-HTP infusion for long term evaluation. Dry matter intake was not affected (P > 0.05) by intra-abomasal infusions. The half-life of 5-HTP was dose-independent (128 min). The serum 5-HTP, serotonin, and 5-hydroxyindoleacetic acid area under the curve increased (P < 0.05) linearly with an increased dose of 5-HTP. Serum 5-HTP reached peak concentration in approximately 30 min after dosing while serum and plasma serotonin peaked after 240 min postinfusion. Serotonin was greater than control for all 5-HTP doses 1 d and 2 d after infusion in serum and plasma, respectively. Intra-abomasal infusion of 5-HTP at doses up to 1 mg/ kg BW increases circulating serotonin for up 2 days., (Copyright © 2021. Published by Elsevier Inc.)

Growing public interest in the use of cannabidiol (CBD) for companion animals has amplified the need to elucidate potential impacts. The purpose of this investigation was to determine the influence of CBD on the daily activity of adult dogs. Twenty-four dogs (18.0 ± 3.4 kg, 9 months-4 years old) of various mixed breeds were utilized in a randomized complete block design with treatments targeted at 0 and 2.5 mg (LOW) and at 5.0 mg (HIGH) CBD/kg body weight (BW) per day split between two treats administered after twice-daily exercise (0700-0900 and 1,700-1,900 h). Four hours each day [1,000-1,200 h (a.m.) and 1,330-1,530 h (p.m.)] were designated as times when no people entered the kennels, with 2 h designated as Quiet time and the other 2 h as Music time, when calming music played over speakers. Quiet and Music sessions were randomly allotted to daily a.m. or p.m. times. Activity monitors were fitted to dogs' collars for continuous collection of activity data. Data were collected over a 14-day baseline period to establish the activity patterns and block dogs by activity level (high or low) before randomly assigning dogs within each block to treatments. After 7 days of treatment acclimation, activity data were collected for 14 days. Data were examined for differences using the MIXED procedure in SAS including effects of treatment, day, session (Quiet or Music), time of day (a.m. or p.m.), and accompanying interactions. CBD (LOW and HIGH) did not alter the total daily activity points ( P = 0.985) or activity duration ( P = 0.882). CBD tended ( P = 0.071) to reduce total daily scratching compared with the control. Dogs were more active in p.m. sessions than in a.m. sessions ( P < 0.001). During the p.m. session, dogs receiving HIGH tended ( P = 0.091) to be less active than the control (CON). During the a.m. and p.m. sessions, CBD reduced scratching compared with CON ( P = 0.030). CBD did not affect the activity duration during exercise periods ( P = 0.143). These results indicate that, when supplemented with up to 4.5 mg CBD/kg BW/day, CBD does not impact the daily activity of adult dogs, but may exert an antipruritic effect., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Morris, Kitts-Morgan, Spangler, Gebert, Vanzant, McLeod and Harmon.)

Although serotonin has been extensively studied in many species, there is a lack of information in ruminants, and no research has been evaluated if its precursor, 5-hydroxytryptophan (5-HTP), administered into the abomasum may be used as a means to manipulate serotonin metabolism. Thus, the objective of this study was to evaluate if intra-abomasal infusion of 5-HTP increases circulating serotonin in the steer. Eight Holstein steers (471 ± 8.9 kg) were used in a replicated 4 × 4 Latin Square design experiment. The treatments were intra-abomasal infusion of 5-HTP at 0.5, 1, 2.5, and 5 mg/kg BW. Blood was collected at 0, 2, 4, 6, 8, and 24 h after infusion. The serum concentration of 5-HTP increased quadratically (P = 0.005) with a peak at 2 h after administration. The 5-HTP administration increased (P < 0.05) serum serotonin in comparison with baseline with no difference (P > 0.05) between the doses of 5-HTP. When 5-HTP was dosed at 2.5 mg/kg BW or higher, intake decreased, and there was an altered manure consistency. The serum 5-hydroxyindole acetic acid concentrations followed the same pattern as 5-HTP. Plasma glucose content was not affected (P > 0.05) by 5-HTP dosing. However, free fatty acids concentration in the plasma was lower (P > 0.05) compared with baseline for the infusion levels of 0.5 and 1 mg/kg BW. Intra-abomasal infusion of 5-HTP efficiently increases serum serotonin cattle., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Serotonin (5-HT) has many important functions in both central and peripheral nervous systems. Although it has been demonstrated that manipulation of serotonin metabolism is possible in many species, there is limited information about l-tryptophan (TRP), a serotonin precursor, in cattle, and these provide conflicting results. Furthermore, there is no study evaluating how different patterns of intra-abomasal infusion of TRP impact circulating 5-HT. The objective of this study was to evaluate if intra-abomasal infusion patterns of TRP can affect circulating 5-HT and other metabolites from TRP metabolism in the plasma and serum and circulating glucose and insulin in cattle. Eight ruminally cannulated Holstein steers were used in a replicated 4 × 4 Latin square design. Each received intra-abomasal water infusion (control) or intra-abomasal TRP infusion (50 mg/kg BW) in 3 different patterns: a pulse infusion once a day (pulse once), pulse infusion twice a day (pulse twice), or continuous infusion (continuous). For continuous treatment, the TRP dose was diluted in tap water and infused by a peristaltic pump (300 mL/h). To equalize conditions, the other treatments had a water infusion (300 mL/h). The steers were fed every 2 h, and blood was collected from a jugular vein catheter every 4 h for 24 h after the initial infusion. Urine produced during the 24 h period was collected. Serum and plasma TRP, 5-HT and kynurenine, plasma glucose, and serum insulin concentrations were analyzed. Urine was analyzed for concentrations of 5-hydroxyindoleacetic acid. Both serum TRP and kynurenine were increased (P < 0.05) by all TRP infusion treatments, but concentrations in pulse dose treatments were greater than those in continuous infusion. Serum 5-HT increased (P < 0.05) with both pulse TRP infusion treatments; however, the continuous TRP infusion did not increase the serum 5-HT. Plasma 5-HT, glucose, and insulin had a tendency to increase with TRP pulse infusions. The urinary 5-hydroxyindoleacetic acid excretion was highest for pulse dose treatments. An acute supply of TRP in 1 or 2 daily doses increases serum 5-HT and increases circulating glucose and insulin in cattle. The TRP and kynurenine concentrations are similar in plasma and serum. However, the serum 5-HT concentration is more responsive to TRP administration than plasma., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Ergot alkaloids can interact with several serotonin (5-hydroxytryptamine [5-HT]) receptors provoking many physiological responses. However, it is unknown whether ergot alkaloid consumption influences 5-HT or its metabolites. Thus, two experiments were performed to evaluate the effect of ergot alkaloid feeding on 5-HT metabolism. In exp. 1, 12 Holstein steers (260 ± 3 kg body weight [BW]) were used in a completely randomized design. The treatments were the dietary concentration of ergovaline: 0, 0.862, and 1.282 mg/kg of diet. The steers were fed ad libitum, kept in light and temperature cycles mimicking the summer, and had blood sampled before and 15 d after receiving the treatments. The consumption of ergot alkaloids provoked a linear decrease (P = 0.004) in serum 5-HT. However, serum 5-hydroxytryptophan and 5-hydroxyindoleacetic acid did not change (P > 0.05) between treatments. In exp. 2, four ruminally cannulated Holstein steers (318 ± 3 kg BW) were used in a 4 × 4 Latin square design to examine the difference between seed sources on 5-HT metabolism. Treatments were: control-tall fescue seeds free of ergovaline, KY 32 seeds (L42-16-2K32); 5Way-endophyte-infected seeds, 5 way (L152-11-1739); KY31-endophyte-infected seeds, KY 31 (M164-16-SOS); and Millennium-endophyte-infected seeds, 3rd Millennium (L108-11-76). The endophyte-infected seed treatments were all adjusted to provide an ergovaline dosage of 15 μg/kg BW. The basal diet provided 1.5-fold the net energy requirement for maintenance. The seed treatments were dosed directly into the rumen before feeding. The experiment lasted 84 d and was divided into four periods. In each period, the steers received seeds for 7 d followed by a 14-d washout. Blood samples were collected on day 0 (baseline) and day 7 for evaluating the treatment response in each period. A 24 h urine collection was performed on day 7. Similar to exp. 1, serum 5-HT decreased (P = 0.008) with the consumption of all endophyte-infected seed treatments. However, there was no difference (P > 0.05) between the infected seeds. The urinary excretion of 5-hydroxyindoleacetic acid in the urine was not affected (P > 0.05) by the presence of ergot alkaloids. In conclusion, the consumption of ergot alkaloids decreases serum 5-HT with no difference between the source of endophyte-infected seeds in the bovine., (© The Author(s) 2020. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Interest is increasing regarding use of Cannabidiol (CBD) in companion animals due to anecdotal evidence of beneficial behavioral and health effects. The purpose of this investigation was to evaluate the influence of CBD on behavioral responses to fear-inducing stimuli in dogs. Sixteen dogs (18.1 ± 0.2 kg) were utilized in a replicated 4 × 4 Latin square design experiment with treatments arranged in a 2 × 2 factorial, consisting of control, 25 mg CBD, trazodone (100 mg for 10-20 kg BW, 200 mg for 20.1-40 kg BW), and the combination of CBD and trazodone. A fireworks model of noise-induced fear was used to assess CBD effectiveness after 7 d of supplementation. Each test lasted a total of 6 min and consisted of a 3 min environmental habituation phase with no noise and a 3 min noise phase with a fireworks track. Plasma was collected 1 h before, immediately after, and 1 h following testing for cortisol analysis. Behaviors in each 3 min block were video recorded, and heart rate (HR) sensors were fitted for collection of HR and HR variability parameters. Research personnel administering treats and analyzing behavioral data were blinded as to the treatments administered. Data were tested for normality using the UNIVARIATE procedure in SAS, then differences examined using the MIXED procedure with fixed effects of treatment, period, time, and treatment x time interaction. Inactivity duration and HR increased during the first minute of the fireworks track compared with 1 min prior ( P < 0.001 and P = 0.011, respectively), indicating the fireworks model successfully generated a fear response. Trazodone lowered plasma cortisol ( P < 0.001), which was unaffected by CBD ( P = 0.104) or the combination with CBD ( P = 0.238). Neither CBD nor trazodone affected the duration of inactivity ( P = 0.918 and 0.329, respectively). Trazodone increased time spent with tail relaxed ( P = 0.001). CBD tended to increase HR ( P = 0.093) and decreased the peak of low- and high-frequency bands (LF and HF, P = 0.011 and 0.022, respectively). These results do not support an anxiolytic effect of CBD in dogs given 1.4 mg CBD/kg BW/d., (Copyright © 2020 Morris, Kitts-Morgan, Spangler, McLeod, Costa and Harmon.)

Pregastric fermentation along with production practices that are dependent on high-energy diets means ruminants rely heavily on starch and protein assimilation for a substantial portion of their nutrient needs. While the majority of dietary starch may be fermented in the rumen, significant portions can flow to the small intestine. The initial phase of small intestinal digestion requires pancreatic α-amylase. Numerous nutritional factors have been shown to influence pancreatic α-amylase secretion with starch producing negative effects and casein, certain amino acids and dietary energy having positive effects. To date, manipulation of α-amylase secretion has not resulted in substantial changes in digestibility. The second phase of digestion involves the actions of the brush border enzymes sucrase-isomaltase and maltase-glucoamylase. Genetically, ruminants appear to possess these enzymes; however, the absence of measurable sucrase activity and limited adaptation with changes in diet suggests a reduced capacity for this phase of digestion. The final phase of carbohydrate assimilation is glucose transport. Ruminants possess Na+-dependent glucose transport that has been shown to be inducible. Because of the nature of pregastric fermentation, ruminants see a near constant flow of microbial protein to the small intestine. This results in a nutrient supply, which places a high priority on protein digestion and utilization. Comparatively, little research has been conducted describing protein assimilation. Enzymes and processes appear consistent with non-ruminants and are likely not limiting for efficient digestion of most feedstuffs. The mechanisms regulating the nutritional modulation of digestive function in the small intestine are complex and coordinated via the substrate, neural and hormonal effects in the small intestine, pancreas, peripheral tissues and the pituitary-hypothalamic axis. More research is needed in ruminants to help unravel the complexities by which small intestinal digestion is regulated with the aim of developing approaches to enhance and improve the efficiency of small intestinal digestion.

Fescue toxicosis is problematic for growing steers, causing lower DMI and productivity when fed endophyte-infected (E+) tall fescue. A complete understanding of underlying mechanisms of how fescue toxicosis affects growing steers is lacking. Therefore, the overall objective of this multiexperiment study was to determine whether ruminally dosed ergovaline (ERV) affects rumen motility, rumen contents, and eating patterns. In Exp. 1, an 8-h period to assess ruminal motility began 4 h after feeding by monitoring pressure changes using a wireless system for 21 d. Eight ruminally cannulated steers (283 kg BW) were pair fed with alfalfa cubes (1.5 × NEm) and assigned to endophyte free (E-; 0 μg ERV/kg BW/d) or E+ treatment (20 μg ERV/kg BW/d). Overall, E+ steers had more frequent rumen contractions (Seed P = 0.05 and day of feeding P = 0.02). On days 7 to 9, both treatments showed lower frequencies and E- steers had greater amplitude of contractions (P < 0.001) that corresponded with decreased DMI. In Exp. 2, steers remained in pairs assigned in Exp. 1 (322 kg BW), but reversed seed treatments while increasing ERV levels (titrated 0, 5, 10, 15, and 20 μg ERV/kg BW/d over 57 d). There were no differences between E- and E+ for frequency (P = 0.137) or amplitude of contractions (P = 0.951), but increasing ERV dosage, decreased frequency (P = 0.018) and amplitude (P = 0.005), coinciding with lower DMI. In Exp. 3, 8 steers (589 kg) were pair fed and ruminally dosed 15 μg ERV/kg BW/d, and rumen motility data were collected for 21 d. E- steers showed higher amplitude and lower frequency of contractions than E+ steers with seed (P < 0.001), day (P < 0.001), and seed × day (P < 0.04) effects, but rumen fill was not different between E- and E+ (P > 0.29). Serum prolactin concentrations were lower in E+ steers in Exp. 1 to 3. Eating patterns of pair-fed E- and E+ steers were relatively slower in E+ than E- (Exp. 4) by measuring every 2 h across 24 h. Number of meals were higher in E+ than E- steers, but meal duration and meal size were not different between treatments. Rumen content (DM%) tended to be higher in E+ than in E- when steers were fed once a day (P = 0.07), but there was no difference for rumen content (DM%) when E- and E+ steers were fed 12 times a day (P = 0.13). These results suggest the changes in rumen fill associated with fescue toxicosis may be driven more by changes in feeding behavior and eating pattern rather than by changes in motility., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We examined the effects of dietary supplementation of a Saccharomyces cerevisiae-based direct-fed microbial (DFM) on the growth performance, whole-blood immune gene expression, serum biochemistry, and plasma metabolome of newly weaned beef steers during a 42 d receiving period. Forty newly weaned Angus crossbred steers (7 d post-weaning; 210 ± 12 kg of BW; 180 ± 17 d of age) from a single source were stratified by BW and randomly assigned to 1 of 2 treatments: basal diet with no additive (CON; n = 20) or a basal diet top-dressed with 19 g of the DFM (PROB; n = 20). Daily DMI and weekly body weights were measured to calculate average daily gain (ADG) and feed efficiency (FE). Expression of 84 immune-related genes was analyzed on blood samples collected on days 21 and 42. Serum biochemical parameters and plasma metabolome were analyzed on days 0, 21, and 42. On day 40, fecal grab samples were collected for pH measurement. Compared with CON, dietary supplementation of PROB increased final body weight (P = 0.01) and ADG (1.42 vs. 1.23 kg; P = 0.04) over the 42 d feeding trial. There was a tendency for improved FE with PROB supplementation (P = 0.10). No treatment effect (P = 0.24) on DMI was observed. Supplementation with PROB increased (P ≤ 0.05) the concentrations of serum calcium, total protein, and albumin. Compared with CON, dietary supplementation with PROB increased (P ≤ 0.05) the expression of some immune-related genes involved in detecting pathogen-associated molecular patterns (such as TLR1, TLR2, and TLR6), T-cell differentiation (such as STAT6, ICAM1, RORC, TBX21, and CXCR3) and others such as TNF and CASP1, on day 21 and/or day 42. Conversely, IL-8 was upregulated (P = 0.01) in beef steers fed CON diet on day 21. Plasma untargeted plasma metabolome analysis revealed an increase (P ≤ 0.05) in the concentration of metabolites, 5-methylcytosine and indoleacrylic acid involved in protecting the animals against inflammation in steers fed PROB diet. There was a tendency for lower fecal pH in steers fed PROB diet (P = 0.08), a possible indication of increased hindgut fermentation. This study demonstrated that supplementation of PROB diet improved the performance, nutritional status, and health of newly weaned beef steers during a 42 d receiving period., (© The Author(s) 2019. Published by Oxford University Press on behalf of the American Society of Animal Science. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Ergot alkaloids from endophyte-infected (Epichloë coenophiala) tall fescue (Lolium arundinaceum) induce vasoconstriction. Previous work has shown that serotonin receptor subtype, 5HT2A, is present in bovine ruminal (R) and mesenteric (M) vasculature, plays a role in vasoconstriction, and could be influenced by ergot alkaloids. To determine the influence of ergot alkaloids on 5HT2A, the vasoactivity of an agonist selective for 5HT2A, (4-bromo-3,6-dimethoxybenzocyclobuten-1-yl) methylamine HCl (TCB-2), was evaluated using bovine ruminal and mesenteric arteries and veins (RA, RV, MA, MV) that were exposed to ergovaline (ERV) prior to or during the TCB-2 additions. Ruminal and mesenteric blood vessel segments were collected, cleaned, and cut into 2- to 3-mm cross-sections. Vessel segments were incubated in Krebs-Henseleit buffer containing 0, 0.01 or 1 µM ERV for 2 h prior to TCB-2 dose response or exposed to ERV concentrations simultaneously during TCB-2 dose response. For the dose response portion of the study, vessels were suspended in a multimyograph containing 5 mL of continuously oxygenated Krebs-Henseleit buffer and equilibrated to 1 g tension for 90 min. Vessels were exposed to increasing concentrations of TCB-2 every 15 min and contractile response data were normalized as a percentage of the maximum contractile response induced by 120 mM KCl reference. Analysis of variance was evaluated separately for each vessel and each ERV exposure experiment using the mixed models procedure of SAS for effects of TCB-2 and ERV concentrations. All blood vessels with previous ERV exposure had significantly lower contractile responses to TCB-2 (P < 0.01). All blood vessels with simultaneous exposure to 1 µM ERV had higher (P < 0.01) contractile responses at lower concentrations of TCB-2. Simultaneous ERV addition at 1 × 10-4 M TCB-2 did not affect contractility of RV, MA, MV (P > 0.05), but decreased contractility of RA (P < 0.01). These results indicate that ergopeptine alkaloid exposure influences contractility of bovine ruminal and mesenteric blood vessels through serotonin receptor subtype 5HT2A by acting as both an agonist and antagonist. Additional work is needed to determine if ergot alkaloids like ERV simply occupy receptor binding sites competitively, or influence receptor internalization to cause the observed divergent responses.

Previous research has demonstrated that exposure to ergot alkaloids reduces vasoactivity of serotonin (5HT) receptors. Chemical suppression of tall fescue seedhead production is a tool to reduce the level of exposure to ergot alkaloids by a grazing animal. Therefore, the objective was to evaluate contractility of lateral saphenous veins biopsied from mixed breed steers following a 87- to 101-d grazing period on 3-ha pastures of bermudagrass (Cynodon dactylon; n = 5 steers; BW = 340 ± 9 kg), or toxic endophyte-infected tall fescue (Lolium arundinaceum) that was not treated (n = 5 steers; BW = 300 ± 6; 0.56 ppm ergovaline) or was treated (n = 5 steers; BW = 294 ± 9 kg; 0.24 ppm ergovaline) with herbicide containing aminopyralid and metsulfuron-methyl. To evaluate contractility, biopsied veins were mounted in a multimyograph and exposed to increasing concentrations of a tall fescue seed extract (EXT; ergovaline source) and 5HT1B (CP 93129), 5HT1D (L-694,247), and 5HT2A (TCB2) agonists. All contractility data were normalized to a maximal response of 1 × 10-4 M norepinephrine and were analyzed as a split plot treatment design using SAS for effects of pasture treatment, agonist concentration, and the interaction. There was no contractile response to any concentration of 5HT1B agonist in any of the pasture treatments. There were pasture × concentration interactions for contractile responses to 5HT2A agonist (P < 0.01) and EXT (P < 0.01). For both EXT and TCB2, veins from bermudagrass steers were more vasoactive to the higher concentrations of these compounds (P < 0.05), and there were no differences between veins collected from the unsuppressed or seedhead-suppressed treatments (P = 0.66). There was also a pasture × concentration interaction for the contractile responses to 5HT1D agonist (P < 0.01). However, these responses were not sigmoidal and reached a zenith at 5 × 10-7 and 1 × 10-6 M. At these concentrations, the response was greatest for veins from the unsuppressed treatment (P < 0.05) and did not differ between veins from suppressed and bermudagrass treatments (P = 0.41). Although reduced levels of ergovaline in seedhead-suppressed pastures did not alter vasoactivity of 5HT2A or 5HT1B receptors in the lateral saphenous vein, elevated vasoactivity of 5HT1D in veins from unsuppressed tall fescue pasture treatment suggests that lower ergovaline levels in seedhead-suppressed pastures can influence the vascular effects of ergot alkaloids.

Prior work using a contractility bioassay determined that the serotonin (5-HT) receptor subtype 5-HT2A is present in bovine lateral saphenous veins and plays a role in ergot alkaloid (EA)-induced vascular contraction in steers grazing endophyte-infected (Epichloë coenophiala) tall fescue (Lolium arundinaceum). Ergot alkaloids have also been shown to be vasoactive in bovine gut vasculature. To determine what 5-HT receptors are involved in vasoconstriction of gut vasculature, contractility of ruminal and mesenteric arteries and veins collected from cattle was evaluated in the presence of agonists selective for 5-HT1B (CP 93129), 5-HT1D (L-694, 247), 5-HT2A (TCB-2), 5-HT2B (BW 723C86), 5-HT4 (BIMU-8), and 5-HT7 (LP 44) receptors. Segments of ruminal and mesenteric veins and arteries were collected and suspended in a multimyograph containing continuously oxygenated Krebs-Henseleit buffer. Blood vessels were exposed to increasing concentrations of 5-HT agonists every 15 min and contractile response data were normalized as a percentage of the maximum contractile response induced by 120 mM KCl. Analysis of variance was evaluated using mixed models procedure of SAS for effects of agonist concentration for each vessel type. Receptor agonists for 5-HT2B, 5-HT1D, and 5-HT7 did not induce a contractile response for ruminal or mesenteric vasculature (P > 0.05). However, when exposed to agonists for 5-HT2B or 5-HT1D, mesenteric veins relaxed below zero (P < 0.05). Exposure of all 4 blood vessel types to 5-HT2A agonist induced contractile responses (P < 0.05). The findings of this study indicate that 5-HT1D and 5-HT2B are present in mesenteric veins and may play a role in vasorelaxation. Further, 5-HT2A is present in ruminal and mesenteric vasculature, plays a role in vasoconstriction of these vessels, and could be influenced by EA exposure as has been demonstrated in peripheral blood vessels.

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). β-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., (© 2017 Blackwell Verlag GmbH.)

Previous research has indicated that serotonergic and α-adrenergic receptors in peripheral vasculature are affected by exposure of cattle grazing toxic endophyte-infected (E+; Epichlöe coenophialia) tall fescue (Lolium arundinaceum). The objective of this experiment was to determine the period of time necessary for the vascular effects of ergot alkaloids to subside. Two experiments were conducted to investigate changes in vascular contractile response and vasoconstriction over time relative to removal from an ergot alkaloid-containing E+ tall fescue pasture. In Experiment 1, lateral saphenous vein biopsies were conducted on 21 predominantly Angus steers (357 ± 3 kg body weight) at 0 (n = 6), 7 (n = 6), 14 (n = 5), or 28 days (n = 4) after removal from grazing pasture (3.0 ha; endpoint ergovaline + ergovalinine = 1.35 mg/kg DM) for 126 days. In Experiment 2, lateral saphenous veins were biopsied from 24 Angus-cross steers (361 ± 4 kg body weight) at 0, 21, 42, and 63 days (n = 6 per time point) following removal from grazing tall fescue pastures (3.0 ha; first 88 days endpoint ergovaline + ergovalinine = 0.15 mg/kg DM; last 18 days endpoint ergovaline + ergovalinine = 0.57 mg/kg DM) for 106 total days. Six steers (370 ± 18 kg body weight) off of bermudagrass pasture for the same time interval were also biopsied on Day 0 and Day 63 (n = 3 per time point). Additionally, in Experiment 2, cross-sectional ultrasound scans of caudal artery at the fourth coccygeal vertebra were taken on Days 0, 8, 15, 21, 29, 36, 42, and 45 to determine mean artery luminal area to evaluate vasoconstriction. In both experiments, steers were removed from pasture and housed in a dry lot and fed a corn silage diet for the duration of biopsies and ultrasound scans. Biopsied vessels used to evaluate vasoactivity were cleaned, incubated in a multimyograph, and exposed to increasing concentrations of 4-Bromo-3,6-dimethoxybenzocyclobuten-1-yl) methylamine hydrobromide (TCB2; 5HT2A agonist), guanfacine (GF; α2A-adrenergic agonist), and (R)-(+)-m-nitrobiphenyline oxalate (NBP; α2C-adrenergic agonist) in both experiments and ergovaline (ERV) and ergotamine (ERT) in Experiments 1 and 2, respectively. In Experiment 1, days off pasture × agonist concentration was not significant (p > 0.1) for all four compounds tested. In Experiment 2, GF, NBP, TCB2 and ERT were significant for days off pasture × agonist concentration interaction (p < 0.02) and vasoactivity increased over time. Vasoactivity to agonists was reduced (p < 0.05) when steers were initially removed from E+ tall fescue pasture compared to bermudagrass, but did not differ by Day 63 for any variable. Luminal areas of caudal arteries in steers grazed on E+ tall fescue relaxed and were similar to steers that had grazed bermudagrass for 36 days on non-toxic diet (p = 0.15). These data demonstrate changes in peripheral vasoactivity and recovery from vasoconstriction occur beyond five weeks off toxic pasture and 5HT2A receptors appear to be more dramatically affected in the lateral saphenous vein by grazing E+ tall fescue pasture than adrenergic receptors., Competing Interests: Mention of trade name, proprietary product of specified equipment does not constitute a guarantee or warranty by the USDA and does not imply approval to the exclusion of other products that may be available.

It was hypothesized that isoflavones may attenuate ergot alkaloid-induced vasoconstriction and possibly alleviate diminished contractility of vasculature after exposure to ergot alkaloids. The objective of this study was to determine if prior incubation of bovine mesenteric vasculature with the isoflavones formononetin (F), biochanin A (B), or an ergovaline-containing tall fescue seed extract (EXT) and their combinations affect ergotamine (ERT)-induced contractility. Multiple segments of mesenteric artery and vein supporting the ileal flange of the small intestine were collected from Angus heifers at slaughter (n = 5, bodyweight = 639 ± 39 kg). Duplicates of each vessel type were incubated in tissue culture flasks at 37°C with a 50-mL volume of Krebs-Henseleit buffer containing: only buffer (control); or 1 × 10(-6) M EXT; F; or B; and combinations of 1 × 10(-6) M EXT + F; 1 × 10(-6) M EXT + B; 1 × 10(-6) M F + B; or 1 × 10(-6) M EXT + F + B. After incubation for 2 h, sections were mounted in a multimyograph chamber. The ERT dose responses were normalized to 0.12 M KCl. Pretreatment with F, B, and F + B without EXT resulted in similar contractile responses to ERT in mesenteric artery and all incubations containing EXT resulted in a complete loss of vasoactivity to ERT. In mesenteric artery pretreated with EXT, treatments that contained B had higher contractile responses (P < 0.05) at ERT concentrations of 1 × 10(-7) and 5 × 10(-7) M. Also, treatments containing B tended (P < 0.1) to have greater responses than treatments without B at ERT concentrations of 1 × 10(-6), 5 × 10(-6), and 5 × 10(-5) M. In mesenteric vein pretreated with EXT, treatments containing F had greater contractile responses to ERT at 1 × 10(-5), 5 × 10(-5), and 1 × 10(-4) M (P < 0.05). These data indicated that F and B at 1 × 10(-6) M and their combination did not impact the overall contractile response to ERT in mesenteric vasculature. However, F and B may offset some of the vasoconstriction caused by prior exposure to ergot alkaloids.

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.

The aim 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 in young, lean, adult dogs. Eight female Beagles were randomly assigned to one of two isonitrogenous and isoenergetic diets, Control or FO, in a crossover design. At the beginning of the experiment and at 30 and 60 d, a baseline blood sample was collected and the dogs then were fed their daily ration. Nitrogen balance began at 07:00 h on day 63 of each experimental period and ended at 07:00 h on day 69. On day 66 of each period, a single dose (7.5 mg/kg) of (15)N-glycine was administered orally to each dog via gelatin capsule. Postprandial glycaemia did not differ between treatments or among sampling days within treatment. Cholesterol concentration was increased (p<0.05) on the Control treatment throughout the experiment when compared to values of day 0. Dogs fed the FO treatment had higher plasma triglyceride and ghrelin concentrations than those fed the Control treatment. Body weight and food intake did not differ between dietary treatments. Faecal excretion was increased (p<0.05) in the FO treatment. Dry matter digestibility was decreased (p<0.05) and fat digestibility tended (p<0.10) to decrease in the FO treatment. Overall, feeding a FO-containing diet showed a protective effect against the rise of plasma cholesterol and it increased plasma ghrelin concentration. However, FO supplementation did not appear to affect protein metabolism or postprandial glycaemia in adult lean dogs.

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 β-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.

The objectives of these experiments were to characterize rumen motility patterns of cattle fed once daily using a real-time wireless telemetry system, determine when to measure rumen motility with this system, and determine the effect of ruminal dosing of ergot alkaloids on rumen motility. Ruminally cannulated Holstein steers (n = 8) were fed a basal diet of alfalfa cubes once daily. Rumen motility was measured by monitoring real-time pressure changes within the rumen using wireless telemetry and pressure transducers. Experiment 1 consisted of three 24-h rumen pressure collections beginning immediately after feeding. Data were recorded, stored, and analyzed using iox2 software and the rhythmic analyzer. All motility variables differed (P < 0.01) between hours and thirds (8-h periods) of the day. There were no differences between days for most variables. The variance of the second 8-h period of the day was less than (P < 0.01) the first for area and less than the third for amplitude, frequency, duration, and area (P < 0.05). These data demonstrated that the second 8-h period of the day was the least variable for many measures of motility and would provide the best opportunity for testing differences in motility due to treatments. In Experiment 2, the steers (n = 8) were pair-fed the basal diet of Experiment 1 and dosed with endophyte-free (E-) or endophyte-infected (E+; 0 or 10 μg ergovaline + ergovalinine/kg BW; respectively) tall fescue seed before feeding for 15 d. Rumen motility was measured for 8 h beginning 8 h after feeding for the first 14 d of seed dosing. Blood samples were taken on d 1, 7, and 15, and rumen content samples were taken on d 15. Baseline (P = 0.06) and peak (P = 0.04) pressure were lower for E+ steers. Water intake tended (P = 0.10) to be less for E+ steers the first 8 h period after feeding. The E+ seed treatment at this dosage under thermoneutral conditions did not significantly affect rumen motility, ruminal fill, or dry matter of rumen contents.

Ergot alkaloids in endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum) have been shown to cause a reduction in blood flow to the rumen epithelium as well as a decrease in volatile fatty acids (VFA) absorption from the washed rumen of steers. Previous data also indicates that incubating an extract of endophyte-infected tall fescue seed causes an increase in the amount of VFA absorbed per unit of blood flow, which could result from an alteration in the absorptive or barrier function of the rumen epithelium. An experiment was conducted to determine the acute effects of an endophyte-infected tall fescue seed extract (EXT) on total, passive or facilitated acetate and butyrate flux across the isolated bovine rumen as well as the barrier function measured by inulin flux and tissue conductance (G t ). Flux of ergovaline across the rumen epithelium was also evaluated. Rumen tissue from the caudal dorsal sac of Holstein steers (n=6), fed a common diet, was collected and isolated shortly after slaughter and mounted between two halves of Ussing chambers. In vitro treatments included vehicle control (80% methanol, 0.5% of total volume), Low EXT (50 ng ergovaline/ml) and High EXT (250 ng ergovaline/ml). Results indicate that there is no effect of acute exposure to ergot alkaloids on total, passive or facilitated flux of acetate or butyrate across the isolate bovine rumen epithelium (P>0.51). Inulin flux (P=0.16) and G t (P>0.17) were not affected by EXT treatment, indicating no alteration in barrier function due to acute ergot alkaloid exposure. Ergovaline was detected in the serosal buffer of the High EXT treatment indicating that the flux rate is ~0.25 to 0.44 ng/cm2 per hour. Data indicate that specific pathways for VFA absorption and barrier function of the rumen epithelium are not affected by acute exposure to ergot alkaloids from tall fescue at the concentrations tested. Ergovaline has the potential to be absorbed from the rumen of cattle that could contribute to reduced blood flow and motility and lead to reduced growth rates of cattle.

Ergot alkaloids are hypothesized to cause vasoconstriction in the midgut, and prior exposure may affect the vasoactivity of these compounds. The objectives of this study were to profile vasoactivity of ergot alkaloids in bovine mesenteric artery (MA) and vein (MV) and determine if previous exposure to endophyte-infected tall fescue seed affected vasoactivity of ergocryptine (ERP), ergotamine (ERT), ergocristine (ERS), ergocornine (ERO), ergonovine (ERN), lysergic acid (LSA), ergovaline-containing tall fescue seed extract (EXT), and 5-hydroxytryptamine (5HT; serotonin). Ruminally cannulated Angus steers (n = 12; BW = 547 ± 31 kg) were paired by weight and randomly assigned to 6 blocks. Steers were ruminally dosed daily with 1 kg of either endophyte-infected (E+; 4.45 mg ergovaline/kg DM) or endophyte-free (E-; 0 mg ergovaline/kg DM) tall fescue seed for 21 d before slaughter. Branches of MA and MV supporting the cranial portion of the ileum were collected after slaughter on d 22, placed in a modified Krebs-Henseleit buffer on ice, cleaned, sectioned, and mounted in a multimyograph chamber. Contractile response was normalized to a maximum KCl response. Inner diameter (P = 0.04) and outer diameter (P = 0.02) of MA were smaller for E+ steers than E- steers. Maximum contractile responses to 120 mM KCl were not different between seed treatments in MA (P = 0.33; E-: 2.67 ± 0.43 g; E+: 3.33 ± 0.43 g) or MV (P = 0.26; E-: 2.01 ± 0.18 g; E+: 1.81 ± 0.18 g). Steers receiving E+ had a smaller (P < 0.01) MA contractile response than E- steers to ERP, ERT, ERS, ERO, ERN, EXT, and 5HT. Steers receiving E+ had a smaller (P < 0.05) MV contractile response than E- steers to ERP, ERT, ERS, ERN, EXT, and 5HT. Lysergic acid failed to induce a contractile response in MA and MV. The contractile response in MA and MV of E- steers produced by 5HT was very large. The EXT was the most potent (P < 0.05) agonist in MV and MA of E+ steers. These data showed that ergot alkaloids were vasoactive in the bovine midgut, and steers exposed to E+ had diminished contractility to some ergot alkaloids in small intestinal vasculature. The findings of this study suggest that dietary exposure to ergot alkaloids has the potential to alter nutrient absorption from the midgut by decreasing blood flow to and from the midgut due to vasoconstriction.

The ergot alkaloid ergovaline has demonstrated a persistent and sustained contractile response in several different vascular models. It was hypothesized that different alkaloids isolated from tall fescue (Lolium arundinaceum) will contribute to this contractile response differently. The objective was to compare contractile-response patterns of single additions of the ergoline alkaloids lysergic acid, lysergol, and ergonovine and the ergopeptine alkaloids ergotamine, ergocristine, ergocryptine, ergocornine, and ergovaline (provided as tall fescue seed extract). Lateral saphenous veins were collected from 6 Holstein steers (BW = 397 ± 28 kg) immediately after slaughter, sliced into cross-sections, and suspended in myograph chambers containing oxygenated Krebs-Henseleit buffer (95% O2/5% CO2; pH = 7.4; 37°C). Treatments were added at 0 min and buffer was replaced in 15-min intervals for a 120-min incubation. In addition to maximum tension and time to reach maximum tension, percent relaxation and rate of relaxation were determined following maximum tension for each treatment. All compounds tested produced significant contractile responses (P < 0.05). All ergoline alkaloids reached maximum response in less time (P < 0.05) than the remaining compounds and began to relax immediately after first buffer change. Lysergic acid had the greatest (P < 0.05) percent relaxation and ergonovine had the greatest (P < 0.05) rate of relaxation. The ergopeptine alkaloids ergovaline, ergotamine, ergocristine, ergocryptine, and ergocornine had slower developing contractile responses with a longer (P < 0.05) interval until maximum tension was achieved compared to the ergoline alkaloids. Maximal responses to all the ergopeptine alkaloids, however, all persisted for the 120-min duration with negligible relaxation occurring. The different classes of alkaloids differed greatly in the type of contractile response generated in the lateral saphenous vein. Persistence of contractile response is thought to be the primary contributing factor to the vasoconstriction observed in animals demonstrating signs of fescue toxicosis, where different ergot alkaloids can contribute differently.

An experiment was conducted to determine if ergot alkaloids affect blood flow to the absorptive surface of the rumen. Steers (n=8) were pair-fed alfalfa cubes and received ground endophyte-infected (Neotyphodium coenophialum) tall fescue (Lolium arundinaceum; E+) seed (0.015 mg ergovaline·kg BW(-1)·d(-1)) or endophyte-free tall fescue (E-) seed via the rumen cannula 2x daily for 7 d at thermoneutral (TN; 22°C) and heat stress (HS; 32°C) conditions. On d 8, the rumen was emptied and rinsed. A buffer containing VFA was incubated in the following sequence: control (CON), 15 μg ergovaline·kg BW(-1) (1×EXT) from a tall fescue seed extract, and 45 μg ergovaline·kg BW(-1) (3×EXT). For each buffer treatment there were two 30-min incubations: a 30-min incubation of a treatment buffer with no sampling followed by an incubation of an identical sampling buffer with the addition of Cr-EDTA and deuterium oxide (D2O). Epithelial blood flow was calculated as ruminal clearance of D2O corrected for influx of physiological water and liquid outflow. Feed intake decreased with dosing E+ seed at HS but not at thermoneutral conditions (TN; P<0.02). Dosing E+ seed decreased serum prolactin (P<0.005) at TN. At HS, prolactin decreased in both groups over the 8-d experiment (P<0.0001), but there was no difference in E+ and E- steers (P=0.33). There was a seed treatment×buffer treatment interaction at TN (P=0.038), indicating that E+ seed treatment decreased reticuloruminal epithelial blood flow at TN during the CON incubation, but the two groups of steers were not different during 1×EXT and 3×EXT (P>0.05). Inclusion of the extract in the buffer caused at least a 50% reduction in epithelial blood flow at TN (P=0.004), but there was no difference between 1×EXT and 3×EXT. There was a seed × buffer treatment interaction at HS (P=0.005), indicating that the reduction of blood flow induced by incubating the extract was larger for steers receiving E- seed than E+ seed. Volatile fatty acid flux was reduced during the 1×EXT and 3×EXT treatments (P<0.01). An additional experiment was conducted to determine the effect of time on blood flow and VFA flux because buffer sequence could not be randomized. Time either increased (P=0.05) or did not affect blood flow (P=0.18) or VFA flux (P>0.80), indicating that observed differences are due to the presence of ergot alkaloids in the rumen. A decrease in VFA absorption could contribute to the signs of fescue toxicosis including depressed growth and performance.

The objective of this study was to validate use of the washed rumen (WR) technique for rapid measurement of fasting heat production (HP) and respiratory quotient (RQ). Sixteen Holstein steers were divided into 2 groups of 8 for a comparison of measurements made during feeding (both groups; 16 steers) and fasting (8 steers; BW = 237 ± 17 kg) and using the WR model (8 steers; BW = 322 ± 30 kg). Steers were maintained in a controlled temperature (21°C) environment and treated as follows: 10 d diet adaptation, 1 d measurement of respiratory gases at 1.5 × NEm (Fed state; all steers d 11), 1 d measurement of respiratory gases under fasting conditions (Fasted; 8 steers d 12) or using the WR technique (8 steers d 12), and 7 d to monitor the reestablishment of intake. Steers were offered alfalfa cubes top-dressed with a mineral premix at 1.5 × NEm. Using an indwelling probe, core temperature (CT) and heart rate (HR) were monitored continuously during the days respiratory gases were measured. For fasting measurements using the WR technique, the reticulorumen was washed and refilled with ruminal buffer (NaCl = 96, NaHCO3 = 24, KHCO3 = 30, K2HPO4 = 2, CaCl2 = 1.5, and MgCl2 = 1.5 mmol/kg of buffer) with Cr-EDTA aerated with 75% N2 and 25% CO2 before introduction to the rumen. Mean hourly CT, RQ, and daily HP between Fasted steers and WR steers were decreased for the WR steers on average from 8 to 24 h after removal of rumen contents (P = 0.049, P < 0.001, and P = 0.076, respectively). Fitting RQ data obtained during fasting to a 1-phase decay equation showed that plateau was achieved at 0.756 ± 0.003 and 0.719 ± 0.003 and time to plateau was 9 and 8 h for Fasted and WR steers, respectively. Mean RQ after WR were 0.778, 0.732, and 0.726 (SEM = 0.003) for time segments 0 to 8 h, 9 to 16 h, and 17 to 24 h, respectively. Mean fasting HP after WR was 18.8, 16.8, and 16.5 (SEM = 0.51) kJ/(h • kg(0.75)) for time segments 0 to 8 h, 9 to 16 h, and 17 to 24 h, respectively. There were no significant differences in RQ and fasting HP (P = 0.23 and P = 0.81, respectively) between the time segment of 9 to 16 h and 17 to 24 h after rumen washing. In contrast, both RQ and HP differed (P = 0.090 and P = 0.081, respectively) across these same time segments for the Fasted group. Therefore, an accurate measurement of fasting HP can be obtained using a shorter-term measurement with the WR technique. This approach provides an alternative to the traditional 48 h fasting time or measurements made during the third and fourth day after starvation.

This study was designed to examine alteration of fasting heat production (FHP) during fescue toxicosis. Six ruminally cannulated Holstein steers (BW = 348 ± 13 kg) were BW-matched into pairs and used in a 2 period crossover design experiment. Each period consisted of 2 temperature segments, one each at 22 and 30°C. During each period, 1 steer per pair was ruminally dosed twice daily with 0.5 kg of ground endophyte-infected fescue seed (E+) and the other with ground endophyte-free fescue seed (E-) for 7 d. Steers on E- treatment were pair-fed to E+ steers offered alfalfa cubes at 1.5 × NEm. On d 8 of each segment, steers were moved to individual metabolism stalls fitted with indirect calorimetry head boxes. Ruminal contents were removed, weighed, and subsampled for DM determinations. The reticulorumen was washed and filled with a buffer (NaCl = 96; NaHCO3 = 24; KHCO3 = 30; K2HPO4 = 2; CaCl2 = 1.5; MgCl2 = 1.5 mmol·kg buffer(-1)) that was gassed with a 75% N2 and 25% CO2 mixture before rumen incubation. During buffer incubation, an E+ or E- fescue seed extract was added at 12 h intervals to maintain treatment presentation to the animal. After a 12-h wait, heart rate, O2 consumption, CO2 production, and urinary output were recorded for 16 h. There was no difference (P = 0.931) in DMI/kg(0.75) between endophyte treatments by design; however, intake decreased (P = 0.004) at 30°C. Increased temperature had no effect (P > 0.10) on other measurements and there were no significant interactions (P > 0.11) of temperature and endophyte treatment. Heart rate was unaffected by fescue treatment or environmental temperature. Percent DM of ruminal contents as well as total rumen DM/kg(0.75) was increased (P < 0.0001) in E+ steers. Respiratory quotient was elevated (P = 0.02) in E+ steers. Oxygen consumption decreased (P = 0.04) and CO2 production tended to be reduced (P = 0.07) during E+ treatment. Calculated FHP (kcal/kg BW(0.75)) was also less (P = 0.006) in steers receiving E+ treatment. These data suggest that consumption of endophyte-infected tall fescue by cattle results in a reduction in basal metabolic rate.