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Fourteen Hereford steers were used to compare carcass traits, meat quality, and fatty acid composition of beef from cattle grazing tall fescue infected with either wild-type (E+; n = 6) or novel, nil ergot alkaloid (AR542; n = 8) endophyte for 209 d. Average daily gain, live weight, and HCW were greater (P < 0.05) for AR542 cattle than for E+. No differences in LM color or pH were observed between AR542 and E+. Steaks from E+ cattle tended (P = 0.10) to have higher L* and b* than those from AR542 cattle at 0 d of display. Ground beef from E+ cattle also had higher (P < 0.05) L* than AR542 cattle, with no differences in a* or b* at 0 d of display. Color changes during display did not differ for both steaks and ground beef from E+ and AR542. Lipid oxidation levels increased (P < 0.05) during simulated retail display, but they did not differ between endophyte treatments. Adipose tissues from E+ cattle had a higher (P < 0.05) percentage of SFA, and a lower (P < 0.05) percentage of MUFA than adipose from AR542 cattle. Ground beef and i.m. fat had higher (P < 0.05) concentrations of SFA, MUFA, and cis-9, trans-11 isomer of conjugated linoleic acid, and lower (P < 0.05) concentrations of PUFA and PUFA:SFA ratio than s.c. fat. The n-6:n-3 fatty acid ratio did not differ among fat depots. Ergot-alkaloids were detected in s.c. adipose tissues, and alkaloid concentration was greater (P < 0.05) for E+ than AR542. Warner-Bratzler shear force values did not differ between endophyte types, but it decreased (P < 0.01) across the postmortem aging period. Conversely, sensory panel evaluation detected greater (P < 0.01) chewiness and lower (P < 0.05) juiciness for AR542 than for E+ steaks aged for 14 d. Although grazing cattle on tall fescue pastures infected with nil ergot alkaloid endophyte improved cattle performance, these results suggest that endophyte type has minor effects on carcass traits and meat quality of pasture-fed beef. Moreover, finishing cattle on tall fescue pastures showed the potential to enhance the fatty acid profile of beef from a human health perspective. Alkaloid concentration was greater (P < 0.05) in s.c. fat from E+ than AR542 (2.81 vs. 0.92 ppb; fresh-tissue basis). This is the first published report demonstrating the presence of alkaloids in beef tissues.

Consumption of wild-type (toxic) endophyte-infected tall fescue (E+) by horses during late gestation is known to adversely affect pregnancy outcome; however, little is known of the potential disruptive consequences of E+ consumption by mares during the critical phases of placentation and fetal development in early pregnancy. The objective of this study was to evaluate the detrimental effects of feeding E+ to mares during early gestation. Mares (n = 12) paired by stage of gestation (d 65 to 100) were assigned to diets (six per diet) consisting of endophyte-free (E-) or E+ tall fescue seed (50% E- or E+ tall fescue seed, 45% sweet feed, and 10% molasses fed at 1.0% of BW/d). Mares also had ad libitum access to E+ or E- annual ryegrass hay, and were fed diets for 10 d. Following removal from the tall fescue diet on d 11, mares were placed on common bermudagrass pasture and monitored until d 21. Morning and evening rectal temperatures were recorded and daily blood samples were collected for progesterone and prolactin (PRL) analyses, whereas samples for 3,4-dihydroxyphenyl acetic acid (a catecholamine metabolite) analysis were collected on alternate days. For clinical chemistry analysis, blood samples were collected on d 0, 5, 10 and 21. Daily urine samples were collected for ergot alkaloid analysis, and ultrasonography was performed for presence of echogenic material in fetal fluids. Rectal temperatures (E+ 37.76+/-0.03; E- 37.84+/-0.03 degrees C) and serum PRL concentrations (E+ 14.06< or =0.76; E- 12.11+/-0.76 ng/mL) did not differ (P = 0.96) between treatments. Measuring the change in basal serum concentration from d 0 over time, progesterone concentrations did not differ (-0.64 +/-1.49 and -0.55+/-1.47 ng/mL for E+ and E- mares, respectively). There was no negative pregnancy outcome, and ultrasonography indicated no increase in echogenic material in fetal fluids. Plasma 3,4-dihydroxyphenyl acetic acid concentrations decreased (P < 0.05) in E+ compared with E- mares (2.1+/-0.14 and 4.4+/0.43 ng/mL, respectively). Urinary ergot alkaloid concentration was greater (P < 0.01) in mares consuming E+ compared with E- (532.12+/- 52.51 and 13.36+/-2.67 ng/mg of creatinine, respectively). Although no fetal loss was observed during the current study, elevated concentrations of urinary ergot alkaloid were consistent with depressed endogenous catecholamine activity, suggestive of an endocrine disruptive effect of hypothalamic origin.

Grazing studies were conducted to determine cattle growth performance, evaluate toxicosis, and compare grazing behavior in stocker cattle grazing nonergot alkaloid-producing endophyte-infected (AR542 or AR502), endophyte-free (E-), or wild-type toxic endophyte-infected (E+) Jesup, Georgia-5, and Kentucky-31 tall fescue. Replicated 0.81-ha tall fescue paddocks were established at the Central Georgia Branch Station at Eatonton and the Northwest Georgia Branch Station at Calhoun during October 1998 and were stocked with beef cattle for autumn and spring periods from fall 1999 through spring 2002. Mean ergot alkaloid concentrations were higher (P < 0.01) on E+ pastures than the other treatments at both locations. At Calhoun and Eatonton, post-treatment serum prolactin concentrations were decreased (P < 0.01) on E+ compared with AR542, AR502, and E- tall fescue. Cattle on AR542, AR502, and E- pastures had lower (P < 0.05) post-treatment rectal temperatures than cattle grazing E+ tall fescue during spring at Eatonton and Calhoun. Calf ADG was higher (P < 0.05) on AR542, AR502, and E- as compared with E+ tall fescue during autumn and spring grazing at Eatonton, and at Calhoun, cattle on E+ pastures had lower (P < 0.05) ADG in both autumn and spring. Gain/hectare was higher (P < 0.05) on AR542, AR502, and E- than on E+ during autumn at Eatonton and during spring at both locations. In autumn at Calhoun, gain/hectare was greater (P < 0.05) on AR502 and E- compared with E+ tall fescue. During April, May, and June, cattle grazing E+ pastures at Eatonton spent more (P < 0.01) time idling, more (P < 0.01) time standing, and used more (P < 0.01) water than cattle on AR542 and E- tall fescue. Daily prehensions and biting rate were each higher (P < 0.01) on AR542 and E- tall fescue than E+ tall fescue in both grazing seasons. There were no differences among pasture treatments for bite size in either spring (P = 0.50) or autumn (P = 0.34). Steers grazing E+ pastures had lower DMI than steers grazing AR542 and E- pastures during spring (P < 0.10) and lower DMI than steers grazing E- pastures during autumn (P < 0.05). Daily steer water usage was decreased (P < 0.10) in E+ pastures compared with AR542 and E- pastures during late fall. These results indicate that nonergot alkaloid-producing endophyte technology is a promising option for alleviating tall fescue toxicosis in stocker cattle.

Nonergot alkaloid-producing endophytes from New Zealand were inserted into tall fescue (Festuca arundinacea) cultivars in an attempt to address the problem of fescue toxicosis in grazing sheep. A 3-yr grazing study was conducted to determine lamb performance and to evaluate toxicosis in lambs grazing nonergot alkaloid-producing endophyte-infected (AR542 or AR502), endophyte-free (E-), or wild-type toxic endophyte-infected (E+) Jesup tall fescue or nonergot alkaloid-producing endophyte-infected (AR542) Georgia-5 tall fescue. Replicated 0.11-ha tall fescue paddocks were established at the central Georgia Branch Station during September 1997 and stocked with lambs from spring 1998 through autumn 2000. Mean ergot alkaloid concentrations were higher (P < 0.01) in E+ forage than in AR542, AR502, and E- tall fescue, and ergot alkaloid concentrations in E- plants and plants infected with AR542 and AR502 were low. Forage availability did not differ (P = 0.92) across treatments during autumn and was higher (P < 0.05) in Georgia-5 AR542 than in Jesup AR502 and E+ pastures. Initial serum prolactin (PRL) concentrations did not differ (P = 0.58) across treatments during autumn, but were higher on Jesup AR542 than E+ during spring. Post-treatment serum PRL concentrations were depressed (P < 0.01) on E+ compared with AR542, AR502, and E- in both spring and autumn. Signs of heat stress were observed in E+ lambs during periods of high ambient temperatures. Mean post-treatment rectal temperature and mean stocking rate exhibited treatment x year interactions (P < 0.05). Lamb ADG was higher (P < 0.05) on AR542, AR502, and E- than on E+ tall fescue. Similarly, gain/hectare was higher (P < 0.015) on AR542, AR502, and E- than on E+. Tall fescue pastures containing AR542 and AR502 endophytes yielded lamb performance that did not differ from that on E- tall fescue and which was superior to performance on E+ tall fescue. Depressed PRL concentrations and elevated rectal temperatures as indicators of toxicosis were evident only in lambs grazing E+ tall fescue, suggesting that nonergot alkaloid-producing endophyte-infected tall fescue is a viable alternative for alleviating tall fescue toxicosis.

Ergot alkaloids cause fescue toxicosis when livestock graze endophyte-infected tall fescue. It is generally accepted that ergovaline is the toxic component of endophyte-infected tall fescue, but there is no direct evidence to support this hypothesis. The objective of this study was to examine relative and potential transport of ergoline and ergopeptine alkaloids across isolated gastric tissues in vitro. Sheep ruminal and omasal tissues were surgically removed and placed in parabiotic chambers. Equimolar concentrations of lysergic acid, lysergol, ergonovine, ergotamine, and ergocryptine were added to a Kreb's Ringer phosphate (KRP) solution on the mucosal side of the tissue. Tissue was incubated in near-physiological conditions for 240 min. Samples were taken from KRP on the serosal side of the chambers at times 0, 30, 60, 120, 180, and 240 min and analyzed for ergot alkaloids by competitive ELISA. The serosal KRP remaining after incubation was freeze-dried and the alkaloid species quantified by HPLC. The area of ruminal and omasal tissues was measured and the potential transportable alkaloids calculated by multiplying the moles of transported alkaloids per square centimeter of each tissue type by the surface area of the tissue. Studies were conducted to compare alkaloid transport in reticular, ruminal, and omasal tissues and to determine whether transport was active or passive. Ruminal tissue had greater ergot alkaloid transport potential than omasal tissue (85 vs 60 mmol) because of a larger surface area. The ruminal posterior dorsal sac had the greatest potential for alkaloid transport, but the other ruminal tissues were not different from one another. Alkaloid transport was less among reticular tissues than among ruminal tissues. Transport of alkaloids seemed to be an active process. The alkaloids with greatest transport potential were lysergic acid and lysergol. Ergopeptine alkaloids tended to pass across omasal tissues in greater quantities than across ruminal tissues, but their transport was minimal compared to lysergic acid and lysergol.

Spectral analysis was used to relate dietary quality and herbage species to the behavior of grazing steers. Four .3-ha paddocks were established with either 'AU-Triumph' tall fescue (F; Festuca arundinacea Schreb.), 'Apollo' alfalfa (A; Medicago sativa L.), 1/3 fescue and 2/3 alfalfa (2/3A), or 2/3 fescue and 1/3 alfalfa (1/3A). Each paddock was stocked with 10 to 16 steers and defoliated in 5 d. Three steers on each paddock carried vibracorders to monitor grazing time. Daily forage samples were taken in 10-cm layers and weighed. Esophageal extrusa were collected from fistulated steers to measure diet quality. Daily grazing time did not differ (P = .37) among treatments; however, steers grazing mixtures grazed numerically longer (1.4 h/d) than steers on monocultures. Spectral analysis revealed that steers grazing A and 2/3A had many daily meals of short duration, but steers grazing 1/3A and F consumed three meals daily at 8-h intervals. Throughout the 4.67-d grazing period, quality of the diet linearly declined in crude protein and herbage digestibility, linearly increased in neutral detergent fiber and cellulose, and exhibited quadratic changes in lignin and ash. For most quality values, the tall fescue monoculture differed from the others (P < .05). Steers selected diets with similar quality for the A, 2/3A, and 1/3A treatments. This study illustrates how differences in forage diets alter grazing behavior of steers.

Three sequential experiments were conducted with rabbits to 1) determine the effect of endophyte-infected (E+) tall fescue seed on rabbit performance and examine the effect of anti-ergot alkaloid immunization on rabbit performance and protectiveness against fescue toxicosis, 2) compare immunogens designed to elicit systemic anti-ergot alkaloid antibodies, and 3) select a superior adjuvant. In Exp. 1, rabbits (n = 6/treatment) fed E+ fescue seed diets (20%, 340 ppb total ergot alkaloids) had reduced (P < .05) intake and weight gain compared with endophyte-free (E-) controls, whereas apparent diet digestibility was not different between E+ and E-. Rabbits immunized against ergot alkaloids (E+ vac) with lysergol conjugated to human serum albumin (Ly-HSA) had greater (P < .05) intake than E+ rabbits during the wk 1 of a 3-wk dietary challenge. In Exp. 2, rabbits (n = 4/treatment) were immunized with Ly-HSA, with H100-B (ergot alkaloid hapten, H100-different protein carrier, B conjugate), or combinations of both with alum as adjuvant. Greatest (P < .001) anti-ergot alkaloid antibody (Ab) titer developed in the group immunized with H100-B. In Exp. 3, rabbits (n = 4/treatment) were immunized with the immunogen H100-B in conjunction with six adjuvants. Freund's incomplete adjuvant (FIA) in combination with DEAE-dextran and FIA alone gave highest anti-ergot titers. In summary, rabbit weight gain and intake were reduced by feeding E+ fescue seed diets, immunization against ergot alkaloids provided temporary improvement in intake, and H100-B conjugate with FIA or FIA + DEAE-dextran as adjuvants elicited a superior anti-ergot immune response. We believe that rabbits may serve as a model animal for fescue toxicosis research.

Ergot alkaloids cause fescue toxicosis when livestock graze endophyte-infected (E+) tall fescue. Little is known about the bioavailability of the ergot alkaloid classes (lysergic acid, lysergic acid amides, or ergopeptine alkaloids) in livestock, and this hampers development of pharmacological strategies to ameliorate the toxicosis. One method used to determine bioavailability of ergot alkaloids is to examine urinary and biliary excretion patterns. Thus, our objectives were to compare ergot alkaloid excretion via urinary or biliary systems and to determine the rate of appearance or clearance of these alkaloids in cattle that were grazing E+ or endophyte-free (E-) tall fescue. In autumn 1996, bile and urine samples were collected from eight steers (203 kg), each grazing E+ and E- tall fescue, and total alkaloid excretion was quantified using competitive ELISA. Approximately 96% of the ergot alkaloids were excreted in urine. The same steers were used to examine the rate of appearance in, or clearance from, urine when switched from E+ to E-, or from E- to E+, pastures in comparison with steers that were continuously grazing E+ or E- tall fescue at 0, 2, 5, and 7 d. Steers were returned to their original pastures after 7 d, and urine was collected at 2, 5, and 7 d. Urinary alkaloid concentrations in steers switched from E- to E+ pastures were similar (P = .55) to those in steers that continuously grazed E+ tall fescue after 2 d. Steers switched from E+ to E- pastures had urinary alkaloid concentrations similar (P = .91) to those in steers that continuously grazed E- pastures after 2 d. In 1997, two trials were conducted in which steers (191 kg) were switched or remained on E+ or E- pastures, and urine was collected at 0, 12, 24, 48, and 96 h to estimate rate of alkaloid appearance or clearance. Steers switched from E- to E+ 1) had about 33% as much urinary alkaloids as steers grazing E+ pasture after 12 h, 2) were not different after 24 h (P = .76), 3) had twice those of the E+ steers at 48 h (P < .05), and 4) were not different after 96 h. Steers switched from E+ to E- tall fescue had approximately 33% less (P < .05) urinary alkaloids than those grazing E+ at 12 h, 67% less (P < .05) at 24 and 48 h, and were not different (P = .86) from steers continuously grazing E- pastures after 96 h. Urinary alkaloid excretion patterns were similar to ergot alkaloid solubility patterns from in vitro digestion of E+ tall fescue. We suggest that alkaloids, liberated from the forage by ruminal microorganisms, were rapidly absorbed as lysergic acid amides and biotransformed ergopeptine alkaloids.