Endophytic fungi (A. coenophialum and A. lolii) which infect grasses produce ergot alkaloids that serve as the grasses' chemical defenses and enhance the vigor of the grass. Turf-type tall fescue with high endophyte levels has been deliberately developed to produce a greener, more vigorous, pest-resistant turf. Consumption of endophyte-infected grass causes various toxicity symptoms in livestock. Cattle in the southeastern and midwestern United States, where tall fescue is grown on 14 million hectares, often develop signs of toxicosis during summer months from grazing plants in fected by A. coenophialum. A more severe form of the disease, fescue foot, has been associated with cold environment and reported in late fall and winter months not only in the southeastern United States but also in the northwest United States. In New Zealand, where perennial ryegrass is grown on 7 million hectares of pasture, sheep often develop a condition called ryegrass staggers from grazing plants infected by A. lolii. New Zealand reports economic losses grazing plants infected by A. lolii. New Zealand reports economic losses associated with the sheep industry of $205 million per year. In the United States, economic losses associated with the beef cattle industry alone is estimated at $600 million per year. Range finding experiments and case studies of fescue foot and perennial ryegrass staggers (PRGS) were conducted on cattle and sheep under grazing and barn conditions. The main objective was to determine threshold levels of the endophyte toxins, ergovaline (EV) (appendix 1) and lolitrem B (appendix 2), associated with the diseases of fescue foot and PRGS respectively. Fescue foot was experimentally induced in cattle under barn studies in the spring with 825 ppb ergovaline. The ergovaline contaminated feed was given for a period of 42 days. Similar barn studies in sheep in spring to early summer did not produce clinical fescue foot with up to 1215 ppb. Field studies of natural fescue foot in a herd of sheep were conducted, (ie 540 ppb) values of ergovaline in the feed, but clinical disease was not produced in late fall through winter. A case study from a herd of sheep revealed 813 ppb dietary ergovaline had produced fescue foot in the months of fall (November). Fields of perennial ryegrass (PRG) where sheep received 2,135 ppb lolitrem B toxin were associated with clinical cases of PRGS in 42 sheep of 237 sheep (18 percent incidence rate) in the Willamette Valley of Oregon. Three months later, sheep on this same field which then had 1,465 ppb lolitrem B, did not have PRGS. These were the first range finding experiments undertaken in this locale to document threshold levels of endophyte toxins associated with fescue foot and PRGS. / Graduation date: 1994

Perennial ryegrass (PRG, Lolium perenne) is a hardy cool-season grass that is infected with the endophytic fungus Neotyphodium lolii, which enables the plant to be insect repellant and drought resistant, lowering the use of insecticides and fertilizers. However, this fungus produces the compound lolitrem B (LB, m/z 686.4) which causes the tremorgenic neurotoxicity syndrome 'ryegrass staggers' in livestock consuming forage which contains <2000 ppb LB. Ergovaline (EV, m/z 534) is a vasoconstrictor normally associated with tall fescue (Festuca arudinacea), but has also been found in endophyte-infected PRG. Past research has shown a strong linear correlation between levels of LB and EV in PRG. The purpose of this study was to examine the linear relationship between EV and LB in feces and determine common metabolites. To accomplish this, four groups of steers (n=6/group) consumed endophyte- infected PRG over 70 days consumed the following averages of LB and EV: group I 2254ppb LB/633 ppb EV; group II 1554ppb LB/ 373ppb EV, group III 1011ppb LB/259ppb EV, and group IV 246ppb LB/<100ppb EV. Group I in week 4 was inadvertently given a washout period at which time the steers consumed the amount of LB and EV given to group IV (control). Both feed and feces samples were extracted using difference solid phase extraction methods and quantified by HPLC-fluorescence for LB and EV. Concentrations of EV and LB obtained through HPLC-fluorescence in both PRG and feces showed a linear relationship. Additional screening for metabolites was conducted LC-MS/MS and showed possible oxidation and reduction metabolites for both toxins. / Graduation date: 2013