F. N. Schrick

Hemodynamics are altered in the caudal artery of beef heifers fed different ergot alkaloid concentrations.

Doppler ultrasonography was used to compare blood flow characteristics in the caudal artery of heifers fed diets with endophyte (Neotyphodium coenophialum) noninfected (E-, 0 microg of ergovaline/g of DM), a 1:1 mixture of endophyte-infected and E- (E+E-; 0.39 microg of ergovaline/g of DM), or endophyte-infected (E+, 0.79 microg of ergovaline/g of DM) tall fescue (Lolium arundinaceum) seed. Eighteen crossbred (Angus x Brangus) heifers [345 +/- 19 kg (SD)] were assigned to individual pens and fed chopped alfalfa hay plus a concentrate that contained E- tall fescue seed for 7 d during an adjustment period. A 9-d experimental period followed with feeding treatments of chopped alfalfa hay plus a concentrate with E+, E-, or E+E- seed being assigned randomly to pens. Doppler ultrasound measurements (caudal artery luminal area, peak systolic velocity, end diastolic velocity, mean velocity, heart rate, and flow rate) and serum prolactin were monitored during the adjustment (3 baseline measures) and during the experimental period (7 measures). Statistical analyses compared proportionate differences between baseline and responses at 3, 27, 51, 75, 171, and 195 h from initial feeding of the experimental diets. Serum prolactin concentrations for E+ and E+E- diets were less (P < 0.001) than baseline concentrations beginning at 27 and 51 h, respectively, from initial feeding of the diets. Although baseline measures were taken when ambient temperatures were likely below thermoneutrality, caudal artery luminal cross-sectional area in E+ heifers had declined (P = 0.004) from baseline by 27 h and remained less (P < 0.02) until 195 h, and caudal artery luminal area declined (P = 0.004) in E+E- heifers from baseline by 51 h and remained less (P < 0.07) until 171 h. Blood flow rate was slower than the baseline rate at 51 h for E+ (P = 0.058) and E+E- (P = 0.02 heifers, but blood flow remained slower in E+E- heifers for 48 h, whereas it remained slower in E+ heifers for 96 h. Adjustments in artery luminal area and blood rate with the 3 diets appeared to parallel the increases in ambient temperature. Heifers fed a diet containing a larger amount of ergot alkaloids had less of a response to ambient temperature than heifers consuming the diet with less or no ergot alkaloids.

Fertility aspects in yearling beef bulls grazing endophyte-infected tall fescue pastures

During a 2-year study, yearling beef bulls were used to determine the effects of grazing on endophyte-infected tall fescue on endocrine profiles, semen quality and fertilisation potential. Bulls were allotted to graze tall fescue pastures infected with Neotyphodium coenophialum (E+; n = 20 per year) or Jesup/MaxQ (Pennington Seed, Atlanta, GA, USA; NTE; n = 10 per year). Bulls were grouped by scrotal circumference (SC), bodyweight (BW), breed composites and age to graze tall fescue pastures from mid-November until the end of June (within each year). Blood samples, BW, SC and rectal temperatures (RT) were collected every 14 days. Semen was collected from bulls every 60 days by electroejaculation and evaluated for motility and morphology. The developmental competence of oocytes fertilised in vitro with semen from respective treatments was determined. Bulls grazing E+ pastures had decreased BW gain (P < 0.01), increased overall RT (P < 0.01) and decreased prolactin (P < 0.01) compared with animals grazing NTE pastures. Neither percentage of normal sperm morphology nor motility differed between bulls grazed on the two pasture types. Semen from E+ bulls demonstrated decreased cleavage rates (P = 0.02) compared with semen from NTE bulls. However, development of cleaved embryos to the eight-cell and blastocyst stages did not differ between the two groups. In conclusion, semen from bulls grazing E+ tall fescue resulted in decreased cleavage rates in vitro, which may lower reproductive performance owing to reduced fertilisation ability.

Ergot alkaloids from endophyte-infected tall fescue decrease reticuloruminal epithelial blood flow and volatile fatty acid absorption from the washed reticulorumen.

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.

Developmental competence of bovine embryos from heat-stressed ova

Because multiple ovulation embryo transfer procedures are occasionally performed in cows experiencing heat stress, the goal of this study was to assess the developmental competence of otherwise morphologically normal embryos from heat-stressed ova. To this end, the ability of compact morulae from heat-stressed and non-heat-stressed bovine ova to undergo blastocyst development after culture at 38.5 or 41.0 degrees C was examined. It was hypothesized that heat-induced perturbations in the ooplasm carry over to increase the susceptibility of the preattachment embryo to heat stress. Initially, ova were matured at 38.5 or 41.0 degrees C. The consequences of heat stress did not include altered cleavage, but did reduce the proportion of 8- to 16-cell-stage embryos (55.3 vs. 50.6%; SEM +/- 1.9). Although proportionately fewer, compact morulae from heat-stressed ova were equivalent in quality to those from non-heat-stressed ova (2.1 and 2.1; SEM = 0.04). Culture of compact morulae from non-heat-stressed ova at 41.0 degrees C did not affect blastocyst development (71.9 and 71.5%; SEM = 3.0). Furthermore, the development of compact morulae from heat-stressed ova was similar to that of non-heat-stressed ova after culture at 38.5 degrees C (68.2 vs. 71.9 and 71.5%; SEM = 3.0). However, blastocyst development was reduced when compact morulae from heat-stressed ova were cultured at 41.0 degrees C (62.3 vs. 71.9, 71.5 and 68.2; SEM = 3.1). In summary, reduced compaction rates of heat-stressed ova explained in part why fewer develop to the blastocyst stage after fertilization. The thermolability of the few embryos that develop from otherwise developmentally challenged ova emphasizes the importance of minimizing exposure to stressor(s) during oocyte maturation.

Dietary exposure to ergot alkaloids decreases contractility of bovine mesenteric vasculature.

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.

Effects of fescue toxicosis on bull growth, semen characteristics, and breeding soundness evaluation.

Tall fescue possesses heat, drought, and pest resistance conferred to the plant by its mutualistic relationship with the ergot alkaloid producing fungal endophyte, Neotyphodium coenophialum. The objective of this study was to evaluate the impact of ergot alkaloid consumption on growth, scrotal circumference (SC), and semen quality. The SC measurement and percentage of motile and normal sperm were used to determine if a bull passed the breeding soundness exam (BSE) requirements. Bulls (n = 14) between 13 and 16 mo of age exhibiting ≥32 cm SC and having passed a BSE were assigned to 1 of 2 dietary treatments accounting for BCS and BW. Bulls were fed the treatment diet containing toxic tall fescue seed (E+; 0.8 μg of ergovaline and ergovalanine/g DM) or the control diet containing endophyte-free nontoxic tall fescue seed (E-) for 126 d. Blood samples were collected and BSE and BCS accessed at the start of the test (d 0) and every 21 d to the end of test (d 126). Weights were obtained on d 0 and d 126. Serum prolactin (PRL) concentrations were affected by treatment × day interactions (P = 0.04) verifying the effectiveness of the E+ diet. Bulls consuming the E+ diet exhibited declining PRL concentrations from 250 ± 52.1 ng/mL on d 0 to 30.6 ± 46.9 ng/mL by d 126 whereas bulls receiving the E- ration maintained serum PRL concentrations greater than or equal to 226.7 ± 50.4 ng/mL across the 126-d study. Body condition score (P = 0.4) and BW (P = 0.4) were not different between treatments. No difference due to treatment was observed for the percentage of bulls passing a standard BSE exam (P = 0.6) and no treatment effect was observed for any semen characteristic measured by computer-assisted semen analysis (CASA; P ≥ 0.2). The SC was negatively affected by treatment × day interaction (P = 0.04) with E- bulls exhibiting a larger SC at d 126 compared with E+ bulls of 36.7 ± 0.8 versus 34.3 ± 0.8 cm, respectively. Within treatment, E+ bulls exhibited a decrease in SC (P = 0.0001) with a d 0 SC of 37.3 ± 0.8 cm and dropping to 34.3 ± 0.8 by d 126. Theoretically, reduced SC would negatively impact semen quality, but this was not observed. However, CASA and BSE evaluation data are consistent with recent reports indicating that bulls grazing E+ tall fescue exhibited only subtle, if any, differences on semen characteristics.

Heat stress effects on the cumulus cells surrounding the bovine oocyte during maturation: altered matrix metallopeptidase 9 and progesterone production

When the effects of heat stress are detrimental during maturation, cumulus cells are intimately associated with the oocyte. To determine the extent to which heat stress affects these cells, in this study, transcriptome profiles of the cumulus that surrounded control and heat-stressed oocytes (41 °C during the first 12 h only and then shifted back to 38.5 °C) during in vitro maturation (IVM) were compared using Affymetrix bovine microarrays. The comparison of cumulus-derived profiles revealed a number of transcripts whose levels were increased (n=11) or decreased (n=13) ≥ twofold after heat stress exposure (P<0.01), sufficient to reduce the development of blastocysts by 46.4%. In a separate study, quantitative PCR (qPCR) was used to confirm heat-induced differences in the relative abundances of the transcripts of five different genes (caveolin 1, matrix metallopeptidase 9, FSH receptor, Indian hedgehog homolog, and inducible nitric oxide synthase). Heat stress exposure resulted in >1.7-fold decrease in the protein levels of latent matrix metallopeptidase 9 (proMMP9). Heat-induced reductions in transcript levels were noted at 6 h IVM with reductions in proMMP9 protein levels at 18 h IVM (P=0.0002). Independent of temperature, proMMP9 levels at 24 h IVM were positively correlated with the development rate of blastocysts (R²=0.36; P=0.002). The production of progesterone increased during maturation; heat-induced increases were evident by 12 h IVM (P=0.002). Both MMP9 and progesterone are associated with the developmental competence of the oocyte; thus, it seems plausible for some of the negative consequences of heat stress on the cumulus-oocyte complex to be mediated through heat-induced perturbations occurring in the surrounding cumulus.

Disparate consequences of heat stress exposure during meiotic maturation: embryo development after chemical activation vs fertilization of bovine oocytes

Consequences of heat stress exposure during the first 12 h of meiotic maturation differed depending on how and when bovine oocytes were activated. If heat-stressed oocytes underwent IVF at ~24 h, blastocyst development was less than for respective controls and similar to that obtained for nonheat-stressed oocytes undergoing IVF at 30 h (i.e. slightly aged). In contrast, if heat-stressed oocytes underwent chemical activation with ionomycin/6-dimethylaminopurine at 24 h, blastocyst development was not only higher than respective controls, but also equivalent to development obtained after activation of nonheat-stressed oocytes at 30 h. Developmental differences in chemically activated vs IVF-derived embryos were not related to fertilization failure or gross alterations in cytoskeletal components. Rather, ionomycin-induced calcium release and MAP kinase activity were less in heat-stressed oocytes. While underlying mechanisms are multifactorial, ability to obtain equivalent or higher development after parthenogenetic activation demonstrates that oocytes experiencing heat stress during the first 12 h of meiotic maturation have the necessary components to develop to the blastocyst stage, but fail to do so after fertilization.

Evaluation of a ruminally dosed tall fescue seed extract as a model for fescue toxicosis in steers.

Tall fescue (Lolium arundinaceum) toxicosis research is often complicated by a reduction in intake of infected forage or seed, making treatment comparisons difficult. This study was conducted to develop a fescue toxicosis model that would allow for variations in DMI without altering the quantity of alkaloids consumed over the course of the experiment. Ground tall fescue seed and a tall fescue seed extract were used in two 2-period crossover experiments to determine the effectiveness of ruminal dosing of a tall fescue seed extract to induce fescue toxicosis. This experiment used 4 growing Holstein steers (BW = 337 ± 24 kg) surgically fitted with ruminal cannulas. Steers were maintained on a diet of endophyte-free fescue hay fed ad libitum throughout the experiment. Endophyte-infected (E+; 4.1 mg/kg of ergovaline) and uninfected (E-; 0.0 mg/kg of ergovaline) KY-31 tall fescue seed was ground and dosed or extracted with ethanol, concentrated, and lyophilized before ruminal dosing. Ergovaline concentration of the final extract was 102 mg/kg. Animals were given a minimum of a 3-wk washout period between treatments. Physiological indicators were measured over 7 d at 22°C (d 1 to 3) and 32°C (d 4 to 7) during both seed and extract dosing. Seed and extract E+ dosing reduced serum prolactin concentrations such that they were not different from zero (P < 0.10). Treatment with E+ reduced feed intake (P < 0.05) and heart rate (P < 0.001), and increased respiration rate (P < 0.01) and core temperature (P < 0.05) during both seed and extract dosing. Increasing environmental temperature from 22 to 32°C reduced total intake (P < 0.05) and increased core temperature (P < 0.001) and respiration rate (P < 0.001) during both seed and extract dosing. Diastolic blood pressure tended (P < 0.09) to be increased during E+ extract dosing and reduced during heat stress. These physiological alterations are consistent with those reported for cattle grazing or consuming seed from endophyte-infected tall fescue. These data indicate that a ruminally dosed ethanol extract of tall fescue seed is efficacious in inducing fescue toxicosis in cattle.

Postgraze assessment of toxicosis symptoms for steers grazed on toxic endophyte-infected tall fescue pasture.

A 2-yr pen experiment was conducted using 12 different crossbred Angus steers each year to determine if short-term changes in prolactin concentrations, body temperature, and vasoconstriction reflect recovery from fescue toxicosis after steers that previously grazed toxic endophyte (Neotyphodium coenophialum)-infected Kentucky 31 tall fescue [Lolium arundinaceum (Schreb.) Darbysh] are placed on nontoxic feed. Groups of 6 steers from toxic endophyte-infected and endophyte-free tall fescue grazing treatments were blocked by BW for assignment to pens as a randomized complete block design with 2 replications. Two environments were implemented by initiating the experiment on 18 August in yr 1 and on 8 September in yr 2 for durations of 30 and 21 d, respectively. Rectal temperatures were recorded, jugular blood was collected for assaying serum prolactin, and cross sections of the caudal artery were ultrasonically imaged at selected time points to evaluate temporal changes in the response variables. Rectal temperatures in steers on the toxic endophyte pasture treatment declined (P < 0.05) linearly over time in yr 1 and 2 and were similar (P > 0.10) to those on endophyte-free treatment on d 30 in yr 1 and by d 15 in yr 2. Prolactin concentrations in steers on the toxic endophyte pasture treatment showed curvilinear increases (P < 0.05) over time and were similar (P > 0.10) to steers on the endophyte-free treatment by d 15 in yr 1 and by d 10 in yr 2. Luminal areas of the caudal artery in toxic endophyte steers were less (P < 0.05) than those in endophyte-free steers across all dates in both years. Results indicated that rectal temperatures in steers after they are removed from toxic fescue may decrease over time, but temporal changes in rectal temperatures could be affected more by prevailing ambient temperatures than by actual mitigation of fescue toxicosis. Prolactin concentrations in steers after they are removed from toxic endophyte tall fescue can increase and stabilize in less than 2 wk, but alkaloid-induced vasoconstriction that causes a vulnerability to severe heat stress is not alleviated within 30 d.