The decrease in productivity caused by fescue toxicosis has been estimated to cost the United States livestock industry more than $1 billion per year due to reduced growth and diminished reproductive efficiency. This goal of the research presented in this dissertation is to enhance the knowledge base concerning the underlying physiological changes that occur during fescue toxicosis that lead to reduced intake and weight gain in cattle. As one of the factors associated with fescue toxicosis is a reduction in feed intake, achieving a consistent and adequate intake of toxins can be a complication. Results from experiment 1demonstrate that ruminal dosing of ground seed and a seed extract are able to mimic the classic symptoms of fescue toxicosis in cattle. This model whereby seed or extract is directly dosed into the rumen eliminates the possibility of reduced alkaloid intake due to refusal of feed by the animal. This model allows for more precise and repeatable dosing of alkaloids in fescue research. Experiment 2 results indicate that ingestion of endophyte-infected tall fescue leads to decreased fasting heat production in cattle. This is indicative of a reduction in maintenance energy requirements and may be related to a decrease in liver size or other metabolic activity in animals grazing endophyte-infected pastures. In addition, a reduction in basal metabolic rate may cause the compensatory gain often observed in cattle entering the feedlot after grazing endophyte-infected pastures. Data from experiment 3 provides evidence that whole body nitrogen and energy metabolism are not altered by fescue alkaloid ingestion. Experiment 3 also addresses the rate of feed degradation and ruminal passage rates in cattle ingesting endophyte infected fescue. While ruminal VFA profile is altered, this is likely due to reduced absorption, not increased production. The data from this experiment indicate that the reduction in weight gain and productivity seen during fescue toxicosis is primarily a function of reduced intake and not secondary effects of alkaloid ingestion.

Ergot alkaloids present in endophyte-infected (E+) tall fescue are thought to be the causative agent of fescue toxicosis, a syndrome affecting cattle in the eastern United States. Many of the observed signs of fescue toxicosis are thought to be attributed to peripheral vasoconstriction; however, there are data indicating that ergot alkaloids can alter blood flow to the gut. An experiment was conducted using right ruminal artery and vein collected from heifers shortly after slaughter. Vessels were mounted in a multi-myograph to determine the vasoconstrictive potentials of ergot alkaloids present in E+ tall fescue. Results indicated ergot alkaloids have the potential to induce vasoconstriction of foregut vasculature. A second experiment was conducted to determine if the additional ergot alkaloids present in E+ tall fescue increase the vasoconstrictive response above that of ergovaline. Results indicated that ergovaline is the main alkaloid responsible for vasoconstriction in bovine vessels. A third study was performed to determine the effect of ergot alkaloids on ruminal epithelial blood flow in the washed rumen of steers exposed to E+ or endophyte-free (E-) tall fescue seed. Steers were dosed with seed followed by a washed rumen experiment with differing levels of ergovaline incubated in the rumen. Results indicated that E+ tall fescue seed treatment reduced ruminal epithelial blood flow. Additionally, incubating ergovaline in the rumen during the washed rumen further decreased epithelial blood flow and volatile fatty acid (VFA) absorption. A final study was conducted to determine the acute effects of ergot alkaloids on isolated rumen epithelial absorptive and barrier functions and the potential for ruminal ergovaline absorption. Results indicate that acute exposure to ergot alkaloids does not alter the absorptive or barrier function of rumen epithelium and ergovaline is absorbed from the rumen. Data from this series of experiments have shown that ergot alkaloids from E+ tall fescue can induce vasoconstriction of blood vessels in the foregut of cattle, reduce blood flow to the rumen epithelium, and decrease VFA absorption. The decrease in nutrient absorption could contribute to the observed symptoms of fescue toxicosis, including depressed growth rates and general unthriftiness.

Objetivou-se avaliar a influência da monensina sódica sobre o acúmulo hepático de cobre em ovinos. Haja vista, que ionóforos podem alterar a absorção, disponibilidade e distribuição tecidual de minerais em muitos tecidos do organismo. Foram utilizados 24 ovinos mestiços das raças Santa Inês x Dorper com média de três meses de idade e peso medio de 30 kg ao início do experimento, que receberam dieta basal calculada em 2,75% do peso corpóreo, sendo 50% de concentrado comercial (7,13 ppm de Cu) e 50% de feno de capim coast-cross (3,72 ppm de Cu) e 10 gramas de sal mineral (5,9 ppm de Cu). Os cordeiros foram distribuídos em quatro grupos distintos, sendo: Controle apenas dieta basal; Monensina (Mon) - dieta basal e 30 ppm de monensina sódica; Cobre (Cu) - dieta basal acrescida de 10 mg/kg P.V./dia de cobre, na forma de sulfato de cobre pentahidratado; e grupo Monensina+Cobre (MonCu) - dieta basal acrescida de 10 mg/kg P.V./dia de cobre e 30 ppm de monensina sódica. O período experimental teve duração de 14 semanas. Foram coletadas amostras de figado e bile no início por biópsia, e ao final do experimento por necropsia, para determinação de minerais, assim como amostras semanais de sangue para avaliação bioquímica, hematológica e mineral. Foi realizada análise estatística por meio de fatorial 2x2 considerando como fatores a inclusão ou não de cobre e monensina na dieta. Somente um animal apresentou quadro clínico de intoxicação cúprica acumulativa. As concentrações de cobre no fígado no início do experimento variaram entre 243,8 e 345,4 ppm não havendo diferença entre os grupos experimentais. As concentrações médias e desvios-padrão em ppm do cobre hepático ao final do experimento foram 3.363 ± 576 para o grupo MonCu; 2560 ± 524 para o grupo Cu; 562 ± 168 para o grupo Mon e 253 ± 131 para o grupo Controle. Ao final do estudo a concentração de cobre hepático foi influenciada pela suplementação com cobre (P = 0,0001) e monensina (P = 0,0003). Os grupos suplementados com cobre tiveram redução dos teores de ferro hepático (P = 0,0287) e aumento das concentrações de cobre na bile. A avaliação bioquimica demonstrou elevação da atividade sérica da GGT e AST (P < 0,05) nos grupos Cu e MonCu a partir da décima primeira semana do experimento em relação aos grupos Controle e Mon. O aumento da atividade destas enzimas foram influenciadas pela suplementação de cobre (P = 0,0340), enquanto os grupos suplementados com monensina apresentaram menores valores de creatinina quinase. Os resultados obtidos permitem concluir que a monensina interfere positivamente com o acúmulo hepático de cobre sendo que a sumplentação deste aditivo pode predispor a quadros de intoxicações cúprica em ovinos. / The aim of this study was to verify if monensin can change the accumulation of cooper in the liver of sheep, in view of ionophores can alter the absorption, availability, tiissue distribution of many minerals in the body. Twenty four crossbred Dorper X Santa Inês lambs wiht mean age of 3 months and wheitgh 30 kg at the begining of trial recived a basal diete calculated by 2,75% of body wheigth of 50% concentrade (7,13 ppm of Cu) and 50% cost-cross hay (3,72 ppm of Cu) and 10 g of mineral salt (5,9 ppm of Cu). Lambs were randomly distributed in four groups: Control: basal diet, Mon Group reciving basal diet plus 30 mg of monensin; Cu group: basal diet plus 10 mg Cu B.W of CuSO4.5H2O solution; Group CuMon: basal diet plus 30 mg of monensin and 10 mg of Cu B.W. The exeperimental period lasted for fourteen weeks. Liver samples were obtined by liver biopsy initially and at the end by necropsy when lambs were slaughterd. Blood samples were obtained weekly for determination of biochemical, hematological and mineral. Statistical analysis using a 2x2 factorial was conducted considering the factors copper and monensin in the diet and the interaction between them to evaluate the hepatic copper at the beginning and at the end of the study. Only one of animal of group MonCu exhibit clinical sings of copper toxicosis. Copper concentrations in the liver at the beginning of the experiment ranged between 243,8 and 345,4 ppm with no difference between experimental groups. Mean concentrations and standard deviations in ppm of hepatic copper at the end of the experiment were 3,363 ± 576 for MonCu group; 2560 ± 524 for Cu group; 562 ± 168 for the Mon group and 2538 ± 131 for the control group. At the end of the study liver concentrations and total hepatic copper accumulation were influenced both by the amount of dietary copper (P = 0.0001) and of monensin (P = 0.0003). The groups supplemented with copper had reduced levels of hepatic iron (P = 0.0287) and increased concentrations of copper in the bile. The biochemical evaluation demonstrated elevated serum AST and GGT activity (P <0.05) with Cu and MonCu from the eleventh week of the experiment compared to the control groups and Mon. The increased activity of these enzymes were influenced by copper supplementation (P = 0.0340), while group monensin had lower values of creatinine kinase. The results indicate that monensin interferes positively with hepatic copper accumulation and supplementation of this additive can predispose cupric poisoning in sheep.

Consumption of endophyte-infected tall fescue results in a syndrome of negatively altered physiological systems, collectively known as fescue toxicosis. Another challenge to endophyte-infected tall fescue -based beef cattle operations is that the soils often are selenium (Se) poor, necessitating the need to provide supplemental Se. To test the general hypothesis that different forms of supplemental Se would ameliorate the negative effects of fescue toxicosis, predominately-Angus steers (BW = 183 ± 34 kg) were randomly selected from herds of fall-calving cows grazing an endophyte-infected tall fescue pasture and consuming vitamin-mineral mixes that contained 35 ppm Se as sodium selenite (ISe), SELPLEX (OSe), or an 1:1 blend of ISe and OSe (MIX). Steers were commonly weaned and depleted of Se for 98 d. Steers were assigned (n = 8 per treatment) to the same Se-form treatments upon which they were raised and subjected to summer-long common grazing of an endophyte-infected tall fescue pasture (0.51 ppm ergot alkaloids: ergovaline plus ergovalinine; 10.1 ha). Selenium treatments were administered by daily top-dressing 85 g of vitamin-mineral mix onto 0.23 kg soyhulls, using in-pasture Calan gates. The first project objective was to determine the effect of forms of supplemental Se on whole blood Se, serum prolactin, liver glutamine synthetase (GS) activity, carcass parameters, and growth performance (Experiment 1). In Experiment 1, whole blood Se increased for all treatments from day 0 to 22 and then did not change. Across periods, MIX and OSe steers had greater whole blood Se than ISe steer. Compared to ISe steers, MIX and OSe steers had more serum prolactin. Liver GS mRNA, protein content, and activity were greater in MIX and OSe steers than ISe steers. However, the ADG and carcass parameters were not affected by Se treatments. The second project objective was to determine the effect of forms of supplemental Se on serum clinical parameters of Experiment 1 steers (Experiment 2). In Experiment 2, across periods, MIX steers had more serum albumin than OSe, and ISe steers, respectively. Serum alkaline phosphatase (ALP) activity was greater in MIX and OSe steers. In addition, blood urea nitrogen (BUN), serum sodium, phosphorus, and magnesium concentration were affected by Se treatments. Partial correlation analysis revealed that serum albumin, BUN, and ALP activity were correlated with whole blood Se concentration. The third project objective was to evaluate the hepatic transcriptome profiles of Experiment 1 steers using microarray and targeted RT-PCR analyses (Experiment 3). In Experiment 3, bioinformatic analysis of microarray data indicated that hepatic glutamate/glutamine, proline, arginine, and citrulline metabolism was affected by different forms of supplemental Se. The mRNA expression of critical proteins involved in glutamate/glutamine (GLS2, GLUD1, GLUL), proline (PYCR1, ALDH18A1), and urea (ARG1, ARG2, OAT, NAGS, OTC, ORNT1) metabolism were differentially expressed by Se treatments. Collectively, we conclude that consumption of 3 mg Se/d as OSe or MIX forms of Se in vitamin-mineral mixes 1) increased whole blood Se content, an indicator of greater whole-body Se assimilation; 2) increased serum prolactin, albumin, and ALP, the reduction of which are hallmarks of fescue toxicosis; and 3) altered hepatic nitrogen metabolism, as indicated by changes in key enzymes of glutamate/glutamine, proline, and urea metabolism. However, 4) these positive effects on metabolic parameters were not accompanied by increased growth performance.

The first goal of the current research was to determine whether gene expression profiles differed between whole pituitaries of growing beef steers grazing pastures containing high (HE) or low (LE) amounts of toxic endophyte-infected tall fescue. The global (microarray analysis) and selected targeted (RT-PCR) mRNA expression patterns of pituitaries collected from beef steers (BW = 266 ± 15.5 kg) that had been randomly assigned to undergo summer-long grazing (89 to 105 d) of either HE (0.52 ppm ergot alkaloids) or LE (< 0.03 ppm ergot alkaloids) pastures were compared. Gene expression data were subjected to one-way ANOVA. The pituitaries of HE steers had 542 differentially expressed genes, and the pattern of altered gene expression was dependent on treatment. Targeted RT-PCR analysis corroborated these findings, including decreased expression of DRD2, PRL, POU1F1, GAL, and VIP and that of POMC and PCSK1, respectively. Canonical pathway analysis (Integrated Pathway Analysis, IPA) identified HE-dependent alteration in signaling of additional pituitary-derived hormones, including growth hormone and GnRH. In conclusion, consumption of endophyte-infected tall fescue alters the pituitary transcriptome profiles of steers in a manner consistent with their negatively affected physiological parameters. The second goal of this project was to test the hypothesis that sodium selenite (ISe), SEL-PLEX (OSe), vs. a 1:1 blend (MIX) of ISe and OSe in a basal vitamin-mineral (VM) mix would differentially alter pituitary transcriptome profiles in growing beef steers (BW = 183 ± 34 kg) commonly grazing an endophyte-infected tall fescue (HE) pasture. Steers were randomly selected from herds of fall-calving cows grazing HE pasture and consuming VM mixes that contained 35 ppm Se as either ISe, OSe, or MIX forms. Steers were weaned, depleted of Se for 98 d, and subjected to summer-long common grazing of a 10.1 ha HE pasture containing 0.51 ppm ergot alkaloids. Steers were assigned (n = 8) to the same Se-form treatments on which they were raised. Selenium treatments were administered by daily top-dressing 85 g of VM mix onto 0.23 kg soyhulls, using in-pasture Calan gates. Pituitaries were collected at slaughter and changes in global (microarray) and selected (RT-PCR) mRNA expression patterns determined. The effects of Se treatment on relative gene expression were subjected to one-way ANOVA. The form of Se affected the expression of 542 annotated genes. Integrated Pathway Analysis found a canonical pathway network between prolactin and POMC/ACTH/ α-MSH synthesis-related proteins, and that mitochondrial dysfunction was a top-affected canonical pathway. Targeted RT-PCR analysis found that the relative abundance of mRNA encoding prolactin and POMC/ACTH/ α-MSH synthesis-related proteins was affected by the form of Se, as were mitochondrial dysfunction-related proteins OSe steers appeared to have a greater prolactin synthesis capacity vs. ISe steers through decreased dopamine receptor D2 signaling, whereas MIX steers had a greater prolactin synthesis capacity and release potential by increasing TRH concentrations than ISe steers. OSe steers also had a greater ACTH and α-MSH synthesis potential than ISe steers. We conclude that form of Se in VM mixes affected genes responsible for prolactin and POMC/ACTH/α-MSH synthesis, and mitochondrial function in pituitaries of growing beef steers commonly grazing an HE pasture. The third goal was to test the hypothesis that sodium selenite (ISe), SEL-PLEX (OSe), vs. a 1:1 blend (MIX) of ISe and OSe in a basal vitamin-mineral (VM) mix would differentially alter selenoprotein profiles in pituitaries and livers of growing beef steers commonly grazing an endophyte-infected tall fescue (HE) pasture (i.e., the same steers used in Goal 2). The effects of Se treatment on relative gene expression were subjected to one-way ANOVA. The mRNA content of 6 selenoproteins in the pituitary was affected by Se treatments, along with two selenoprotein P receptors, whereas the expression of two selenoproteins was altered in the liver. We conclude that the change in selenoprotein gene expression in pituitaries indicates that OSe steers have a greater potential capacity to manage against oxidative damage, maintain cellular redox balance, and have a better quality control of protein-folding in their pituitaries than ISe steers. The change in selenoprotein gene expression by the liver indicates that MIX steers have a greater redox signaling capacity and capacity to manage oxidative damage than ISe steers.

Where lead pollutes urban soils, both human and animals risk exposure. This exposure gives rise to similar health risks across species. A group of 50 outdoor living cats from inner city Tampa, Florida were tested for blood lead concentration (BLC). Most cats had no measurable lead loads, 14 percent had levels less than or equal to 6 micrograms per decaliter. Soil samples were taken from the home location of each cat. None of these samples, which ranged from 2.6 microgram per gram to 170 micrograms per gram, had hazardous levels of lead. Overall, BLCs were lower than expected. In addition, the BLCs were lower than those found in older industrial cities. The reduction of the use of lead as well as Tampa's location in the newly developed Sunbelt, may be responsible for the overall low levels found in the region's outdoor living cat population.

A wild-type endophyte (Neotyphodium coenophialum [(Morgan-Jones & Gams) Glen Bacon & Hanlin]) that infects tall fescue [Lolium arundinaceum (Schreb.) Darbysh. = Schedonorous arundinaceum (Schreb.) Dumort.] imparts tolerances to moisture, heat, and grazing stresses, but also produces ergot alkaloids that adversely affect performance and physiology of cattle. Novel endophytes, developed by AgResearch Ltd. NZ, can sustain fescue persistence and productivity, but do not produce toxic ergot alkaloids. University of Kentucky Plant Breeder, T. D. Phillips Ph.D, developed a tall fescue experimental population (KYFA9301) for the upper transition zone. A 2-yr grazing experiment was conducted with steers to evaluate steer performance and physiology, and forage quality and productivity of KYFA9301 infected with AR584 novel endophyte (AR584) compared with KY31 wild-type endophyte (KY31), endophyte-free KYFA9301 (EF9301) and AR542-‘Jesup’ (MaxQ). Fescue-endophyte combinations were assigned to 1.0-ha pastures in a randomized complete block design with three replications. Pastures were grazed with variable stocking (four testers) from 6 May to 23 July 2008 (76 d), and 2 April to 25 June 2009 (84 d). Shrunk bodyweights were taken at initiation and termination of grazing each year. Average daily gains among MaxQ, AR584, and EF9301 were similar and were greater (P < 0.10) than KY31. Rectal and skin temperatures were collected three times each year at approximately days 28, 56, and study completion, along with blood collection for serum prolactin assay. Rectal and skin temperatures among AR584, MaxQ, and EF9301 were similar and were lower (P < 0.10) than KY31. Serum prolactin concentrations were similar among the three nontoxic varieties and higher (P < 0.10) than KY31. Forage collections were taken at 2 week intervals throughout the study each year and nutritive quality analysis were conducted through wet chemistry to determine forage acid detergent fiber (ADF), neutral detergent fiber (NDF), crude protein (CP), and In-Vitro Dry Matter Digestibility (IVDMD). Nutritive analyses indicated no differences between fescue-endophyte combinations with the exception of EF9301 having higher ADF concentrations (P = 0.031) than KY31 during the dry year of 2008. Results indicated steer performance and physiological responses for KYFA9301, with and without AR584 were enhanced compared to KY31 and similar to those for MaxQ.

Twenty-four steers were grazed on endophyte (Neotyphodium coenophialum)-infected [[Lolium arundinaceum (Schreb.) Darbysh]; E+] tall fescue (TF) and exposed to ergot alkaloids for an 106-d grazing period. Cattle were removed from pasture, placed in dry lots, and fed a non-toxic diet to evaluate changes in vascular contraction relative to time-off E+ TF pasture. Lateral saphenous veins (SV) were biopsied from steers at 0-, 21-, 42-, and 63-d off TF pasture and from 6 control steers at 0- and 63-d off bermudagrass (BG) pasture. To evaluate contractile response, biopsied SV were exposed to increasing concentrations of ergotamine in a multimyograph. Cross-sectional scans of the caudal artery were taken using color Doppler ultrasonography on 0-, 8-, 15-, 21-, 29-, 36-, 42-, and 45-d to determine artery luminal area. Contractility of SV was less for TF than BG steers on d 0, but was similar between the two treatments by d 63. Luminal areas of the caudal arteries in E+ TF steers relaxed over time and were similar to BG steers by 36 d off pastures. Results indicated that alkaloid-induced vasoconstriction in cattle grazed on E+ TF can be relaxed in 5 to 6 weeks after they are placed on non-toxic diets.

Fescue toxicosis can produce negative effects on animal weight gain and physiology. Sixty-four steers were grazed on endophyte-infected (E+) KY-31 tall fescue for 77 days in 2007 and sixty steers grazed for 86 days in 2008 to evaluate interactions with implantation of steroidal implants and concentrate feeding on performance and physiology of yearling steers. Steers were stratified by body weight for assignment to six, 3.0-ha toxic tall fescue pastures. The main plot treatment of with or without pelleted soybean hulls (SBH) were randomly assigned to pastures. Pelleted SBH were group-fed to provide daily consumptions of 2.3 kg/steer/d (as fed). Sub-plot treatments of with or without ear implantation with steroid hormone (200 mg progesterone--20 mg estradiol were assigned to groups of five or six steers within each pasture. Average daily gain in the experiment showed an additive effect of feeding SBH and implanting (P

Endophyte-infected (E+) tall fescue grass has been associated with fescue toxicosis, a costly syndrome characterized by poor cattle performance and health resulting in significant production losses. The fungal endophyte produces ergot alkaloids, which help the grass thrive in poor conditions but are toxic to mammals. A number of symptoms of fescue toxicosis can be related to vasoconstriction of bovine core, peripheral, and foregut vasculature. The first part of this series of experiments demonstrated ergot alkaloids were also vasoactive in midgut vasculature, with the exception of lysergic acid. Additionally, prior dietary exposure to ergot alkaloids decreased the contractile response of mesenteric vasculature to many of the ergot alkaloids tested. In the second part of this series, a non-invasive method was developed for measuring rumen motility in cannulated cattle. Using this technology without different dietary treatments, it was determined that 8 to 16 h after feeding was the least variable between animals and would provide the best opportunity to measure differences in motility. Application of this technique in the third part of this series investigated the effect of ruminally dosed ergot alkaloids on rumen motility. Treatments were not effective at inducing fescue toxicosis, and no differences in rumen motility variables were detected.