Volatile fatty acid metabolism by epithelial cells isolated from different areas of the ewe rumen

Cells were harvested from four rumen locations in four 2- to 3-yr-old ewes fed fescue hay to determine whether cell origin has an effect on cellular VFA metabolism. Tissue (approximately 150 cm2) was excised from the anterior cranial pillar, ventral sac floor, caudal pillar surface, and dorsal sac ceiling. Cells were isolated using serial tryptic digestion. One milliliter of isolate was incubated for 2 h in 6 mL of medium containing 25 mM propionate and 10 mM butyrate. Incubations were terminated at 0, 30, 60, 90, and 120 min and analyzed for beta-hydroxybutyrate, acetoacetate, lactate, and pyruvate. Cell yield was 22, 22, 24, and 14 (+/- 6) x 106 cells/mL, and viability was 92, 92, 94, and 87% for anterior cranial pillar, ventral sac floor, caudal pillar surface, and dorsal sac ceiling, respectively. All metabolite concentrations and ratios of redox pairs increased throughout the incubations, indicating continuous cellular activity. Final 2-h concentrations (nmol/10(6) cells) were 123, 113, 163, and 158 (+/- 35) for beta-hydroxybutyrate; 38, 42, 24, and 45 (+/- 10) for acetoacetate; 25.3, 20.6, 10.1, and 20.4 (+/- 5.6) for lactate; and 2.54, 0.98, 1.06, and 1.31 (+/- 0.61) for pyruvate in the anterior cranial pillar, ventral sac floor, caudal pillar surface and dorsal sac ceiling incubations, respectively. Origin of rumen tissue had no significant effect on metabolite production, indicating that cellular location is not a critical factor that affects rate of rumen epithelial cell VFA metabolism under these specific in vitro conditions.     

Ability of induced corpora lutea to maintain pregnancy in beef cows

Experiments were conducted in beef cows without a primary CL, in which pregnancy had been maintained with exogenous progestogen. In preliminary trials, replacement CL induced ipsilateral to the embryo and after, rather than before, d 36 of pregnancy, maintained more pregnancies after withdrawal of exogenous progestogen (13/13 vs 2/6; P < 0.05). In Exp. 1, in cows with replacement CL induced by treatment with hCG on d 28 of pregnancy, treatment with flunixin meglumine on d 31 through 37 did not increase maintenance of pregnancy. Experiment 2 was conducted to evaluate directly the effects of concentrations of PGF2alpha and estradiol-17beta during d 31 through 35 of pregnancy on maintenance of pregnancy by replacement CL induced between d 28 and 31. In cows that maintained pregnancy while progestogen was provided, maintenance of pregnancy after withdrawal of exogenous progestogen tended to be greater with high (5/5) than with low (2/6; P < 0.10) concentrations of PGF2alpha and greater with low (6/7) than with high (2/6; P = 0.10) concentrations of estradiol-17beta. Secretion of progesterone by replacement CL was greater (P < 0.05) in cows with high than in those with low concentrations of PGF2, during d 31 through 35. Prostaglandin F2alpha may facilitate attachment of the bovine embryo (d 30 to 40) in a manner similar to that reported for implantation in other species. Cows that did not form CL in response to hCG on d 28 to 31 responded well when retreated after d 36. Again, maintenance of pregnancy was greater when replacement CL were induced after (9/9) rather than before d 36 (8/16; P < 0.05).     

Maintenance of pregnancy in postpartum beef cows that have short-lived corpora lutea.

The first two experiments examined the role of the uterus in low pregnancy rates of beef cows induced to ovulate by early weaning. At 20 to 25 d postpartum, one-half of the cows in Exp. 1 and 2 received a s.c. implant containing 6 mg of norgestomet (NOR) for 9 d (NOR-pretreated) and the remaining cows were untreated controls (CON). Lengths of first postpartum luteal phase after weaning of calves at d 7 after implant insertion were expected to be normal in NOR-pretreated and short in CON cows. In Exp. 1, cows of both groups received an implant containing 3 mg of NOR at d 4 after first estrus and a silastic implant with 15 or 25 mg of NOR at d 7 after first estrus. At 7 d after first estrus, two embryos were transferred into the uterus of each cow and pregnancy was diagnosed by ultrasonography at d 35. Blood samples were collected daily from onset of treatment to d 8 after estrus and then every other day to d 24. Only 4 of 22 cows were pregnant at d 35, concentrations of estradiol (E2) were elevated after luteolysis, and large follicles were present at d 35. In Exp. 2, all cows were injected with 100 mg of progesterone (P4) twice daily from d 4 to 35 after first estrus. Embryos were transferred, pregnancy was diagnosed, and blood samples were collected as in Exp. 1, except blood sampling was continued to d 34.(ABSTRACT TRUNCATED AT 250 WORDS)     

Is nutritional anestrus precipitated by subfunctional corpora lutea in beef cows?

Thirty-four multiparous, lactating, cyclic beef cows which calved in moderate body condition were used to determine effects of restricted nutrition on corpus luteum (CL) development and endocrine status. At 78 d postpartum, six cows were assigned to a control (CON) diet (26.0 Mcal ME), fed to increase bodyweight (BW) and body condition score (BCS), and the remaining 28 cows were fed to lose BW and BCS on a restricted (RES) diet (14.0 Mcal ME). Following a 40-d adjustment period on respective diets, estrous cycles were synchronized and cows bled daily for determination of progesterone (P4), luteinizing hormone (LH) and insulin (INS) beginning at the synchronized estrus. Ultrasonography was used to determine the ovulatory follicle and CL development. Control cows were maintained for one estrous cycle and were ovariectomized on day 11 of their second cycle. Ten cows on restricted diet (RES-C) continued to form a functional CL (P4 > 1.5 ng/ml at day 10 of an estrous cycle) through as many as 5 cycles, after which observations were discontinued. Fourteen cows on restricted diet (RES-A) were ovariectomized on day 11 of a cycle when a CL was identified by ultrasonography, but was subfunctional (P4 < 1.5 ng/ml on day 10 of that cycle). Four additional RES-A cows which had subfunctional CL were not ovariectomized but were bled for an additional 25 d. At ovariectomy, CL and ovarian weights were collected. Luteal tissue was prepared for evaluation of P4 synthesis, LH responsiveness in vitro, and for determination of P4 content and total LH receptors. Bodyweight and BCS increased in CON cows; whereas, RES cows lost BW and BCS (P < .05). In the cycle prior to ovariectomy, serum P4 and LH were not different in 18 RES-A cows which developed subfunctional CL in comparison to CON cows. Four RES-A cows not ovariectomized but bled for an additional 25 d neither exhibited estrus, ovulated, nor had P4 concentrations greater than .3 ng/ml. Serum INS was lower in RES-A cows during the cycle prior to ovariectomy than in CON cows (P < .05). During the 11-d period prior to ovariectomy, mean serum P4 and INS were lower in RES-A cows than in CON cows (P < .05); however, serum LH was not different. Furthermore, CL and ovarian weights, P4 content of CL, secretion of P4 by luteal tissue in response to LH in vitro and LH receptor number were not different between CON and RES-A cows. In conclusion, nutritional anestrus may be preceded by the formation of a CL with lower steroidogenic output in vivo. However, luteal tissue, collected from RES-A cows, did not appear to be subfunctional during in vitro incubation when substrate availability and gonadotropin support were equal between diets.     

Ovulation and estrus characteristics in crossbred Brahman heifers treated with an intravaginal progesterone-releasing insert in combination with prostaglandin F2alpha and estradiol benzoate.

Crossbred Brahman heifers (n = 60) were studied to determine the effect of a 7-d intravaginal progesterone-releasing insert (INSERT) in combination with PG (Lutalyse; 25 mg i.m.) and estradiol benzoate (EB; .5 mg i.m.) on time of ovulation and estrous behavior. In Phase I, heifers at unknown stages of the estrous cycle were assigned by BW and body condition score to one of the three treatments on d 0: 1) INSERT for 7 d and PG on d 7 (CONTROL; n = 10); 2) INSERT for 7 d, PG on d 7, and EB 24 h after INSERT removal (EB24; n = 10); or 3) INSERT for 7 d, PG on d 7, and EB 48 h after INSERT removal (EB48; n = 10). Blood samples were collected every 8 h after INSERT removal. Also, blood sampling and ultrasonography began 8 h after the onset of estrus, determined with HeatWatch devices, and every 4 h thereafter to detect ovulation. In Phase II, Phase-I treatments (n = 10/treatments) were replicated, but only behavioral estrus data were collected to minimize handling of heifers. Frequent handling of heifers did not influence (P > .1) the interval from INSERT removal to the onset of HeatWatch and visual estrus and duration of estrus, so behavioral estrus data were combined for Phases I and II. Interval from INSERT removal to HeatWatch estrus was decreased (P < .05) in EB24 (45.5 h) vs EB48 (55.9 h) and CONTROL (59.2 h). Interval from INSERT removal to ovulation differed (P < .04) between CONTROL, EB24, and EB48 (93.5, 74.5, and 78.9 h, respectively). Ovulatory follicle size was similar (P > .1) between CONTROL, EB24, and EB48 (14.4, 12.5, and 14.1 mm, respectively). Duration of estrus was similar for CONTROL, EB24, and EB48 (14.0, 15.1, and 17.6 h, respectively). No difference (P > . 1) was observed in number of mounts received between CONTROL, EB24, and EB48 (28.0, 25.7, and 39.4, respectively), but number of mounts received increased in Phase II vs Phase I (40.0 and 22.2, respectively; P < .05). In conclusion, EB hastened the interval from INSERT removal to ovulation without altering duration of estrus or number of mounts received. Frequent handling of heifers did not affect interval to first mount received after INSERT removal or duration of estrus, but it decreased the total number of mounts received.     

The alarmin interleukin-33 drives protective antiviral CD8⁺ T cell responses

Pathogen-associated molecular patterns decisively influence antiviral immune responses, whereas the contribution of endogenous signals of tissue damage, also known as damage-associated molecular patterns or alarmins, remains ill defined. We show that interleukin-33 (IL-33), an alarmin released from necrotic cells, is necessary for potent CD8(+) T cell (CTL) responses to replicating, prototypic RNA and DNA viruses in mice. IL-33 signaled through its receptor on activated CTLs, enhanced clonal expansion in a CTL-intrinsic fashion, determined plurifunctional effector cell differentiation, and was necessary for virus control. Moreover, recombinant IL-33 augmented vaccine-induced CTL responses. Radio-resistant cells of the splenic T cell zone produced IL-33, and efficient CTL responses required IL-33 from radio-resistant cells but not from hematopoietic cells. Thus, alarmin release by radio-resistant cells orchestrates protective antiviral CTL responses.     

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.