Cholinergic reactivity of the bovine fetal ruminal wall in vitro

Strips of rumen wall from bovine fetuses were incubated in an organ bath with acetylcholine only (0.16 to 5.12 g/ml) or in the presence of neostigmine (0.20 g/ml) or atropine (0.05g/ml). The highest reactivity was observed in the period of 4.0–5.9 months of fetal age. This reactivity could be associated with the starting point of rumen papillary development.     

Field trial of a planned breeding regimen for dairy cows, using gonadotrophin-releasing hormone and prostagladin F2alpha

A planned breeding regimen, using gondadotrophin-releasing hormone (GnRH), prostaglandin F2alpha, and a second dose of GnRH, followed by a fixed time insemination, was evaluated in comparison with a negative control on eight commercial dairy farms in the south of England. Fertility data were collected from the 220 cows in the planned breeding group and from 220 matched control cows inseminated at observed oestrus. The planned regimen induced visible oestrus in the vast majority of the cows, and serving the cows at this oestrus reduced the calving to conception interval by 15 days, resulting in 12 per cent more cows being pregnant by 125 days after calving, and 6 per cent more by 150 days. The results from the individual farms suggested that more benefit may be derived from using the regimen in herds with only average fertility indices. There was also evidence to suggest that the second GnRH injection was important, even in cows that came on heat and were served before the fixed time insemination.     

Effect of vitamin E on ruminal fermentation in vitro

The effects of vitamin E on pH value, total protozoa counts, volatile fatty acid (VFA), ammonia nitrogen and lactate levels were examined using an in vitro ruminal incubation system. The ruminal fluid (100 ml) of the first and second group was supplemented with 0.4 mg or 0.8 mg of vitamin E, respectively. Samples were taken immediately before and following 3, 6, 12 and 24 h of incubation at 39 degrees C and analysed for the total protozoa counts, the pH and the levels of ammonia nitrogen, lactate and VFA. Levels of propionate at 24 h and ammonia nitrogen at 12 and 24 h were significantly higher in the second group than in the control. In contrast, the levels of butyrate at 6, 12 and 24 h and lactate at 6, 12 and 24 h were lower in the second group than in the control. Propionate at 24 h, acetate levels at 6, 12 and 24 hand ammonia nitrogen levels at 6, 12 and 24 h and total rumen protozoa counts at 6, 12 and 24 h were significantly higher in the second group as compared with control. In contrary, butyrate levels at 6, 12 and 24 h, lactate levels at 6, 12 and 24 h were lower in second group than in control. There was no statistically significant difference among the groups in the pH values. In conclusion, the addition of vitamin E to in vitro ruminal fluid was found to increase the concentrations of acetate and propionate, total counts of protozoa, levels of ammonia nitrogen, but to decrease the butyrate and lactate levels of the ruminal aliquots in in vitro ruminal fermentation.     

Prostaglandin F2alpha (PGF2alpha) independent and dependent regulation of the bovine luteal endothelin system

We have examined the genes of the endothelin system that are targets for regulation by prostaglandin F2alpha (PGF2alpha). The effects of a luteolytic dose of PGF2alpha ) on the mRNA encoding endothelin converting enzyme-1 (ECE-1), pre-pro endothelin-1 (pp ET-1) and the ET receptors ETA, ETB, in bovine corpus luteum (CL) during the early (days 1 and 4), mid (day 10) or late (day 17) luteal phases were examined. The effect of the PGF(2alpha) treatment on ECE-1 protein, Big ET-1 and the biologically active mature ET-1 peptide were also examined. Most importantly, the direct ECE-1 activity was determined. Before day 10 of the cycle, in a PGF2alpha-independent manner, the amounts of mRNA encoding ET-1, ECE-1, ETA, and ETB were increased steadily from day 1. After day 10 of the cycle, expression of mRNA encoding pp ET-1 and ETA acquired responsiveness to exogenous PGF2alpha and both genes were up-regulated by the PGF2alpha treatment. This effect of PGF2alpha was also detected for the proteins corresponding to the mature ET-1. The enzymatic activity of ECE-1 remained unchanged throughout the lifespan of the CL in spite of the detected changes in mRNA and protein. The results suggest that the luteal endothelin system is regulated in a PGF2alpha-independent and -dependent manner. Importantly, an alteration in luteal ET-1 availability is most likely achieved by modulating the expression of mRNA encoding pp ET-1 and not by the amount or activity of ECE-1. This interpretation is supported by the observation that the activity of ECE-1 remained unchanged throughout the ovarian cycle. The combined effects of greater ET-1 availability and gene expression encoding the ETA receptor in the late luteal phase could render the CL, at this developmental stage, more sensitive or responsive to ET-1. If the luteal tissue is responsive to the available ET-1 during the early phase of the ovarian cycle, an additional role for ET-1 should be considered beyond mediating the luteolytic actions of PGF2alpha. Agents blocking the actions of ET-1 might be the best approach to interfere with the luteal ET system and test its physiological role(s) in vivo.     

Factors that determine rates of cyanogenesis in bovine ruminal fluid in vitro

Strained ruminal fluid was collected from cattle fed five diets at two locations to determine in vitro rates of cyanogenesis from the glycosides amygdalin, prunasin and linamarin. Rates of dissociation for the corresponding aglycones, benzaldehyde cyanohydrin and acetone cyanohydrin, also were determined. Hydrogen cyanide (HCN) in ruminal fluid was determined with a modified method of HCN analysis that independently measured the overall rate of cyanogenesis and the nonenzymatic dissociation of cyanohydrins, the intermediate products in the degradation of cyanogenic glycosides to HCN. Rate of dissociation of cyanohydrins in ruminal fluid was pH-dependent, with high rates of dissociation (as expressed by the rate constant or half-life of the reaction) occurring at pH greater than 6 and slower rates at pH 5 to 6. Cyanohydrin dissociation was most rapid when cattle were fasted for 24 to 48 h and ruminal pH was high; rate of dissociation was much slower during feeding and digestion. When the glycosides were examined, highest rates of cyanogenesis (mg HCN.liter-1.s-1) were observed after a 24-h postprandial period. Hence, cattle are most susceptible to poisoning by cyanogenic plants when the pH of ruminal fluid is elevated (for rapid dissociation) and also when the activity of microbial beta-glucosidase is adequate for rapid hydrolysis of glycosidic bonds. Rates of cyanogenesis were higher when ruminal inocula were from cattle fed fresh alfalfa or cubed alfalfa hay rather than grain or long hay. Rates of HCN production were slowest using inocula from cattle fed grain; rates for the three glycosides were negligible at the 3 and 6 h postprandial sampling times.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of prostaglandin F2 alpha administration on early pregnancy in ewes

Two trials were conducted with ewes to determine the effects of prostaglandin F2 alpha (PGF) administration during the first week of gestation. In trial 1, ewes (n = 134) were checked for breeding activity once daily and half of them received 10 mg PGF im at either 0, 24, 48, 72, 96, 120 or 144 h after detection of a breeding mark. The other half served as uninjected controls. In trial 2, ewes (n = 153) were checked for breeding activity twice daily. Two-thirds of the ewes received 10 mg PGF at either 24, 36, 48, 60, 72, 84, 96, 108, 120 and 132 h following detection of a breeding mark. The other one-third of the ewes served as uninjected controls corresponding to treatment times of 24, 48, 72, 96 or 120 h. In trial 1, the percentage of ewes lambing as a result of first service decreased as time of administration of PGF increased. The first-service pregnancy rate was 87.5% for ewes given PGF at 0 h and 0% for ewes given PGF at 144 h. Fewer (P less than .05) ewes given PGF at 96, 120 or 144 h after first mating lambed than control ewes. Similarly in trial 2, fewer (P less than .05) ewes given PGF at 96, 108, 120 or 132 h after first mating lambed than did controls. The total number of ewes lambing as a result of the entire breeding season did not differ (P greater than .05) between treated and control ewes in trial 1 (88.2 vs 87.3%) or trial 2 (85.7 vs 83.3%).(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of prostaglandin F2 alpha on adrenal-produced steroid hormones in cows

Ovariectomized, nonlactating cows were treated with IM injections of either physiologic saline solution or prostaglandin F2 alpha. Plasma concentrations of cortisol increased significantly by 30 to 60 minutes after injection of prostaglandin F2 alpha, but there were no significant increases in plasma concentrations of estradiol, progesterone, or testosterone. After saline solution treatment, there were no increases in any of the hormones measured.     

Effect of fetal bovine serum and heat-inactivated fetal bovine serum on microbial cell wall-induced expression of procoagulant activity by equine and canine mononuclear cells in vitro

OBJECTIVE: To determine the effect of fetal bovine serum (FBS) and heat-inactivated FBS (HI-FBS) on lipopolysaccharide (LPS)- and zymosan-induced procoagulant activity of equine and canine mononuclear cells. SAMPLE POPULATION: Mononuclear cells from 18 horses and 3 dogs. PROCEDURES: Cells were incubated with various concentrations of FBS, HI-FBS, LPS, zymosan, polymyxin B, and anti-LPS-binding protein monoclonal antibody or combinations of these constituents. A 1 stage recalcification assay was used to determine procoagulant activity. RESULTS: Addition of FBS to media significantly increased procoagulant activity; equine and canine cells were stimulated by 1% and 10% FBS, respectively. Coincubation of cells with FBS and polymyxin B did not reduce this effect, suggesting that the response was not attributable to LPS contamination. Addition of HI-FBS to media did not stimulate procoagulant activity of equine or canine cells, and the sensitivity of the equine cells to LPS was significantly increased by HI-FBS. This increased LPS sensitivity was reduced 40% with monoclonal antibody directed against human recombinant LPS-binding protein. Increasing concentrations of HIFBS significantly increased LPS- and zymosan-induced procoagulant activity of canine cells. CONCLUSION AND CLINICAL RELEVANCE: Procoagulant activity production in equine and canine mononuclear cells was significantly increased by addition of FBS, whereas heat inactivation of FBS eliminated this effect. Heat inactivation did not eliminate the function of serum proteins involved in enhancement of LPS and zymosan-induced procoagulant activity. Results suggest that HI-FBS can be used as a source of serum proteins that increase the sensitivity of mononuclear cells to bacterial and yeast cell wall components.     

Effect of prostaglandin F2 alpha on the fertility of dairy cows after calving

Three hundred and thirty-one dairy cows in six herds were used to study the effect of a single injection of 25 mg dinoprost, a prostaglandin F2 alpha derivative, administered eight days after calving on several indices of fertility. A detailed comparison was made between 113 treated cows and 113 control cows of the same parity, which had calved within seven days of each other on the same farm and had experienced the same degree of dystocia. In cows which had calved without assistance, the treatment reduced the mean interval between calving and first heat from 40 days to 37 days but increased the mean interval between calving and conception from 83 days to 85 days. In cows which had required assistance at calving, the treatment reduced the mean interval between calving and first heat from 44 days to 34 days and reduced the mean calving to conception interval from 86 days to 68 days. The cows which benefited most were those which had required assistance during their second, third or fourth calvings; their calving to conception intervals were shortened by 22.5 days. An average of 1.39 services was required to establish conception in the treated cows which calved unaided, with 70 per cent conceiving to the first service, compared with an average of 1.34 services per conception and a 72 per cent conception rate to first service for their untreated herdmates.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of ergot alkaloids on contractility of bovine right ruminal artery and vein

Ergot alkaloids produced by the endophyte (Neotyphodium coenophialum) associated with tall fescue (Lolium arundinaceum) are implicated in the clinical signs of fescue toxicosis. These compounds were hypothesized to correspondingly affect foregut vasculature. The objective of this study was to determine vasoconstrictive potentials of ergovaline, ergotamine, ergocryptine, ergocristine, ergonovine, ergocornine, and lysergic acid on right ruminal artery and vein. Segments of right ruminal artery and vein were collected from the ventral coronary groove of predominantly Angus heifers (n = 10) shortly after slaughter and placed in a modified Krebs-Henseleit buffer on ice. Vessels were cleaned of excess connective tissue and fat, sliced into 2- to 3-mm segments, and suspended in a multi-myograph chamber with 5 mL of continuously oxygenated Krebs-Henseleit buffer (95%O(2)/5% CO(2); pH 7.4; 37°C). Arteries and veins were equilibrated to 1.0 and 0.5 g, respectively, for 90 min followed by the reference addition of 120 mM KCl. Increasing concentrations of each alkaloid were added to the respective chamber every 15 min after buffer replacement. Data were normalized as a percentage of the contractile response induced by KCl. Alkaloid (P < 0.0001), concentration (P < 0.0001), and vessel type (artery or vein; P = 0.004) affected contractility. No arterial response was observed until 10(-6) M for ergovaline and ergotamine; 10(-5) M for ergocryptine, ergocornine, and ergonovine; and 10(-4) M for ergocristine. Lysergic acid did not induce a contractile response in the ruminal artery. No venous contractile response was observed until concentrations of 10(-6) M for ergovaline, 10(-5) M for ergotamine, and 10(-4) M for ergocryptine and ergocristine were achieved. Lysergic acid, ergonovine, and ergocornine did not induce a contractile response in the ruminal vein. A greater arterial maximal response was observed for ergovaline (P < 0.0001), whereas the arterial and venous responses were not different for ergotamine (P = 0.16), ergocryptine (P = 0.218), and ergocristine (P = 0.425). These results indicate that ergot alkaloids associated with toxic endophyte-infected tall fescue are vasoactive and can potentially alter arterial blood supply and venous drainage from the bovine foregut.     

Effect of prostaglandin F2 alpha on hormone concentrations in dairy cows after parturition

This study was designed to evaluate the effects of exogenous prostaglandin F2 alpha (PGF2 alpha) on hormone secretion in cows without a corpus luteum. Blood samples were taken from 10 Friesian dairy cows at frequent intervals from a jugular vein and the caudal vena cava starting between nine and 20 days after parturition. PGF2 alpha (25 mg dinoprost) was injected intramuscularly into five cows after the first eight hours of sampling. Plasma concentrations of 13,14-dihydro 15-keto PGF2 alpha (PGFM) increased rapidly but had returned to baseline by 14 hours after injection. There was no significant effect of the treatment on the time taken by the cows to resume ovarian cycles, and it had no consistent effect on plasma luteinising hormone (LH) patterns; however the amplitude of pulses of LH was temporarily suppressed in two cows and the frequency of pulses of LH was immediately increased in one cow. Treatment with PGF2 alpha had no significant effect on the concentration of oestradiol in blood from the vena cava. It is concluded that any enhancement of the reproductive performance of cows treated with PGF2 alpha after parturition is not due to a direct effect on pituitary-ovarian function.     

Effect of prostaglandin F2 alpha, phenylephrine and ergonovine on uterine contractions in the ewe

Two experiments were conducted to determine the effect of prostaglandin F2 alpha (PGF2 alpha), phenylephrine and ergonovine on uterine contractions. In the first experiment, ewes were bilaterally ovariectomized, and a strain gauge force transducer was sutured to the serosa of one uterine horn. Each ewe was treated sc with 2 micrograms of estradiol-17 beta daily to prevent regression of the uterus. Beginning at least 5 d after ovariectomy, four dose levels of PGF2 alpha, phenylephrine and methoxamine were given by im injection and ergonovine was given by im or iv injection. Phenylephrine, methoxamine and ergonovine are alpha-adrenoceptor agonists. Uterine activity was recorded by physiograph for 30 min before and 90 min after treatment. Tracing were analyzed for 20-min periods before treatment and 4 to 24 min and 50 to 70 min after treatment. In Exp. 2, transducers were attached to uteri of intact ewes at d 10 to 12 of an estrous cycle. During subsequent estrus, one or two dose levels of PGF2 alpha, phenylephrine and ergonovine were given by im injection and uterine activity recorded. In Exp. 1, PGF2 alpha and phenylephrine increased (P less than .05 or .01) the number of amplitude of contractions at both 4 to 24 and 50 to 70 min. Ergonovine given im increased the number of contractions. In intact estrous ewes, PGF2 alpha increased the number and amplitude of contractions at 4 to 24 min, phenylephrine increased the number and amplitude at both 4 to 24 and 50 to 70 min, and ergonovine increased the number slightly but significantly at 4 to 24 min.(ABSTRACT TRUNCATED AT 250 WORDS)     

Changes in bovine luteal progesterone metabolism in response to exogenous prostaglandin F(2alpha)

An experiment was conducted to determine the effect of prostaglandin F2alpha (PGF2alpha) on luteal synthesis of progesterone (P4) and related progestins. Sixteen beef heifers were assigned in equal numbers to four groups in a 2x2 factorial arrangement of treatments. The experiment consisted of two levels of PGF2alpha analog (0 and 500 microg) and two levels of time (4 and 24 h after injection) of corpus luteum collection. All heifers were injected intravenously with saline (2 ml) or PGF2alpha (cloprostenol) on day 8 of the estrous cycle (estrus=day 0). Jugular blood was collected at 0, 1, 2, 3, 4 and 20, 21, 22, 23, and 24 h after injection. Resulting sera were analyzed for P4 by use of radioimmunoassay. Luteal tissue was analyzed by gas chromatography/mass spectrometry for P4, 20beta-hydroxyprogesterone, pregnenolone, and allopregnanolone (3beta-hydroxy-5alpha-pregnan-20-one). Treatment with PGF2alpha reduced serum concentrations of P4 as early as 1 h after injection (P<0.005) and steroid levels remained low over 24 h. Similarly, administration of PGF2alpha caused a decline in luteal P4 (P<0.005), 20beta-hydroxyprogesterone (P<0.10), and pregnenolone (P<0.05). In contrast, treatment with PGF(2alpha) caused an increase in luteal allopregnanolone over time (time x treatment interaction; P<0.05). These data are interpreted to suggest that PGF2alpha promotes conversion of P4 to the metabolite allopregnanolone.     

Effects of prostaglandin F(2alpha) and nitric oxide on the secretory function of bovine luteal cells

The objective of the present study was to investigate the influence of prostaglandin F(2alpha) (PGF (2alpha)) and nitric oxide (NO) on production of steroids and PGs by culturing bovine luteal cells obtained from ovaries on days 8-12 of the estrous cycle with a nitric oxide (NO) donor (Spermine NONOate), and a NO synthase inhibitor (N(G)-nitro-L-arginine methyl ester dihydrochloride: L-NAME). When the cells were exposed for 24 h to PGF(2alpha) (10(-7)-10(-5) M), production of progesterone (P(4)) increased significantly at all doses used (P<0.05). Moreover, PGF(2alpha) stimulated PGF(2alpha) production (P<0.01), depressed testosterone (T) production (P<0.05), but did not affect synthesis of prostaglandin E(2) (PGE(2)). Spermine NONOate decreased P(4) production to 66%, 47% and 34% of the control concentration after treatment with 10(-5) M, 10(-4) M and 10(-3) M, respectively, but did not affect T production, and increased PGF(2alpha) synthesis (P<0.05) and PGE(2) (P<0.01) at all doses used. L-NAME increased production of P(4) (P<0.01) but did not affect (P>0.05) secretion of T, PGF(2alpha) and PGE(2). Estradiol-17beta (E(2)) was detectable on the level of sensitivity of assay and was not significantly altered by any treatments. The overall results suggest that PGF(2alpha) and NO produced locally in bovine CL play roles in the regulation of the secretory function of the bovine CL as auto/paracrine factors.     

Effect of administrating oxytocin or prostaglandin F2alpha on characteristics of the canine ejaculate

To test the hypothesis that male dogs treated with smooth muscle contracting drugs have an increase in the total number of spermatozoa in the ejaculate but no change in all other ejaculate characteristics, such as progressive motility of spermatozoa or percentage morphologically normal spermatozoa, dogs were treated with oxytocin or prostaglandin F2alpha (PGF2alpha) and compared to saline treatments. Semen was collected from each of the 3 dogs once every 3 to 4 d for a total of 6 collections per dog. Ten minutes before each collection, 1 of 3 injections (oxytocin 10 IU [0.5 mL], IM; PGF2alpha 2.5 mg [0.5 mL], IM; or saline 0.5 mL, IM) was administered. Compared to the saline controls, neither treatment had any significant effect on any measured variable when collected in this manner with an estrus bitch present. Therefore, the use of these drugs does not appear to be a viable treatment to increase the number of spermatozoa.