Selenium (Se) has received a lot of attention for its antioxidant and immune modulating properties. Yet, comparably few studies have focused on the horse. Therefore the objectives of this research were to evaluate the influences of Se status on immune function and antioxidant defense in horses. Twenty eight horses were allocated to one of 4 dietary Se treatments: low (LS), adequate (AS), high organic (SP) and high inorganic (SS). First, horses assigned to LS, SP and SS were depleted of Se and received a low Se diet (0.07 ppm Se) for 35 wk, while AS received an adequate Se diet (0.14 ppm Se). During week 28 to 35 immune function was evaluated using a vaccine challenge with keyhole limpet hemocyanin (KLH) and equine influenza as antigens. Then, a 29 wk repletion phase followed. The LS and AS received the same diets described above while SP received an organic Se supplemented diet (0.3 ppm; Sel-Plex, Alltech, Nicholasville, KY) and SS an inorganic Se supplemented diet (0.3 ppm; sodium selenite). Immune function was assessed using a vaccine challenge with ovalbumin (OVA) and equine influenza as antigens during week 22 to 29. Samples collected throughout the depletion and repletion phases were used to assess change in Se status, antioxidant status and oxidative stress. Finally, a mild exercise test served to assess exercise induced oxidative stress. The experimental model responded as hypothesized, evaluated by blood Se and glutathione peroxidase (GPx) activity. Upon vaccination with KLH, antibody response was faster in AS than LS. Antigen specific mRNA expression of T-bet was also higher for AS than LS. Following OVA vaccination humoral and cell-mediated vaccination responses were similar across treatments. However, non-specific stimulation of peripheral blood mononuclear cells indicated suppressed mRNA expression of selected cytokines for LS compared to AS, SP and SS. Antioxidant capacity and oxidative stress were unaffected by change in Se status. A difference in GPx response post exercise was also noted between SP and SS. Low Se status impaired some measures of immune function. Supplementation at 0.3 ppm may benefit horses as indicated by higher GPx activity in idle and exercised horses.

The hormone glucagon-like peptide-2 (GLP-2) is important in the regulation of intestinal growth and blood flow in nonruminant animals. However, no research reports the existence of GLP-2 in ruminants. Therefore, this dissertation examined the existence of GLP-2 and its receptor, their response to physiological stimuli, and its ability to induce gastrointestinal growth and intestinal blood flow in ruminants. Experiments 1 and 2 established the gastrointestinal distribution of mRNA for proglucagon (the GLP-2 precursor) and the GLP-2 receptor. Furthermore, these experiments determined the effects of changing dietary energy intake on plasma GLP-2 concentrations and proglucagon and GLP-2 receptor mRNA expression. Experiment 3 examined the effect of exogenous bovine GLP-2 on splanchnic blood flow, splanchnic nutrient flux, and gastrointestinal growth. This research shows that ruminants possess a functional GLP-2 signaling system that responds to nutrient ingestion. Based on observed receptor distribution and growth changes with GLP-2 treatment, GLP-2 targets the small intestine and does not affect forestomach or large intestinal growth. Increases in ileal proglucagon mRNA expression and plasma GLP-2 with increasing energy intake demonstrate that GLP-2 responds to physiologic changes in nutrient intake and can be relevant to feeding practices. Furthermore, observed increases in small intestinal growth and blood flow with GLP-2 suggest that it could substantially affect the capacity of the gastrointestinal tract for nutrient absorption. Modification of GLP-2 through diet could allow for improvements in nutrient utilization and animal productivity. This research also has important implications for use of GLP-2 for human disease therapy as the observed downregulation in the blood flow response to 10-d GLP-2 administration has never been reported in any species prior to this dissertation. This research systematically characterized and evaluated the potential role of GLP-2 in the control of gastrointestinal growth and splanchnic blood flow in ruminants. While it extends the knowledge of hormonal control of the gastrointestinal tract in ruminants, it also adds crucial information to the larger body of work investigating the actions of GLP- 2. This dissertation research has contributed to the groundwork necessary to enable the use of GLP-2 in improving the health and productivity of a diverse group of mammalian species.

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.

Two experiments were conducted at the University of Kentucky, Eden Shale Farm, Owenton, KY to evaluate the use of MTB-100TM (Alltech, Inc., Nicholasville, KY) to alleviate the symptoms of fescue toxicity when lactating Angus x Beefmaster cows and their calves grazed endophyte-infected KY-31 tall fescue. Experiment 1 provided a carbohydrate based toxin adsorbent, MTB-100TM, ad libitum in a commercial mineral supplement to project a daily consumption rate of 0, 20 or 40 g of MTB-100TM per cow. Cows were weighed, assigned a body condition score (BCS) and hair coat score (HC), rectal temperatures were recorded and fecal grab samples were taken for ergovaline (EV) and lysergic acid (LA) analysis every 35 days for three grazing seasons (May to September). Calves were also weighed and assigned a HC score. Although MTB-100TM did not improve cow or calf performance, cows older than 4 years and those with greater Beefmaster breeding exhibited a higher tolerance to fescue toxicity than 2 and 3-yr-olds and cows with greater Angus breeding. Experiment 2 was conducted to evaluate the response of lactating beef cows and their calves to strategic supplementation with MTB-100TM. MTB-100TM was mixed with a complete mineral so daily intake was projected to be 0 or 20 g/cow. The experimental period extended from May 5 to October 2 and was divided into 3 strategic periods: P1 = May 5 to July 5; P2 = July 5 to August 31; P3 = August 31 to October 2. Treatments were either 0 or 20 g•cow-1•d-1 MTB-100TM within a period (Treatment 1 = 0, 0, 0; Treatment 2 = 20, 0, 20; Treatment 3 = 0, 20, 0; Treatment 4 = 20, 20, 0; and Treatment 5 = 20, 20, 20). Cow and calf performance was measured the same as Exp. 1, but every 21 days. Supplementation early in the grazing season tended to improve cow weight gain and body condition; however, there was no effect on calf performance. Fecal output of EV and LA did not increase in either experiment with supplementation. In conclusion, strategically invoked MTB-100TM consumption can increase performance of cows grazing endophyte-infected tall fescue forage.

Parascaris equorum is a nematode parasite that infects young horses, sometimes causing unthriftiness, respiratory signs, or intestinal impaction in severe cases. Infection can be diagnosed by detection of eggs in feces, but this is only possible after the worms are fully mature. The goal of this study was to develop an antibody-based test for prepatent diagnosis of P. equorum infection. To produce western blot (WB) antigen, P. equorum larvae were cultured for collection of excretory-secretory antigens (ESA). Sera from 18 pregnant broodmares, their subsequent foals, and a group of 12 older mares and geldings were analyzed. In order to check for cross-reactivity between P. equorum and other ascarid species and equine parasites, additional sera were analyzed. Sera from a horse with monospecific P. equorum infection was compared to horses with monospecific Strongyloides westeri or cyathostome infections, rabbits inoculated with Baylisascaris procyonis or Toxocara canis eggs, dogs naturally infected with T. canis, and rabbits immunized with B. procyonis or P. equorum ESA. Molecular weights of silver-stained P. equorum larval ESA ranged between 12 to 94 kDa. In WB analysis, sera from 94% of broodmares contained IgG(T) antibody that recognized multiple P. equorum larval ESA. Foals showed no IgG(T) antibodies pre-suckle, but antibodies similar to their dams were observed post-suckle and thereafter. Of the older mares and geldings, 58% had IgG(T) antibodies recognizing larval ESA. Serum IgG(T) antibodies against P. equorum larval ESA were also found in parasite-free and monospecific infection equine sera. Ascarid positive foals did not produce detectable amounts of IgE or IgM antibodies against larval ESA. When P. equorum, T. canis, and B. procyonis antibody reactivity was compared, antigens at 19 kDa and 34 kDa had the highest potential for identification of larval P. equorum infections. When immature adult P. equorum ESA was examined, IgG(T) antibody recognition was demonstrated in 50% of broodmares and 17% of the older horses, and appeared several weeks prior to patency in foal serum. Results indicate that IgG(T) antibodies against P. equorum ESA are common in mature horses, and are transferred from mare to foal, limiting the diagnostic potential of an antibody-based test.

Placental insufficiency-induced intrauterine growth restriction (IUGR) increases risk of mortality and morbidity in newborn infants and domestic animals. IUGR fetuses are typically exposed to prolonged hypoxemia, hypoglycemia, and hypercatecholaminemia, which results in perinatal pancreatic β-cell dysfunction. Recent evidence indicates that chronic exposure to norepinephrine in utero suppresses insulin secretion through α2-adrenergic receptors (ARs), but if the adrenergic actions are blocked compensatory hyper insulin secretion response is observed in the IUGR sheep fetus. In the current studies, we demonstrate that chronic NE exposure alone can produce the compensatory enhancement of β-cell responsiveness following termination of a chronic NE infusion. In the fetus NE was continuously infused at 1-4 μg/min for seven days starting at 131 days of gestational age (term = 145 days). During treatment, NE infused fetuses had higher (P < 0.05) plasma NE concentrations and lower (P < 0.01) insulin concentrations than vehicle infused control fetuses. Glucose stimulated insulin secretion (GSIS), which measures β-cell function, prior to NE treatment was not different between treatments. However, insulin concentrations during hyperglycemic steady state period of GSIS studies and area under the curve of glucose-potentiated arginine-induced insulin secretion were higher (P < 0.01) than control values and this augmentation was confirmed at 3 hours, 24 hours, and five days in NE-infused fetuses after discontinuing the infusion. Pancreatic islets isolated within 10 hours post NE infusion had lower (P < 0.05) mRNA expression of α1D (58%), α2A (43%), α2C (42%), α1 (67%) adrenergic receptors (ARs), and uncoupling protein 2 (40%) compared to islets from controls. Isolated islets from NE-infused fetuses 5 days after NE treatment had lower (P < 0.05) inhibitory responsiveness from NE and a greater (P < 0.05) maximal insulin release with glucose simulation in static incubations compared to controls. These findings show that following chronic NE exposure insulin secretion responsiveness was augmented and was coupled with desensitized adrenergic signaling. Moreover, this compensatory β-cell enhancement persists for days indicating chronic NE exposure permanently alters β-cell responsiveness.

<p>Making decisions is fundamental to everything we do, yet it can be impaired in various disorders and conditions. While research into the neural basis of decision-making has flourished in recent years, many questions remain about how decisions are instantiated in the brain. Here we explored how primates make abstract decisions and decisions in social contexts, as well as one way to non-invasively modulate the brain circuits underlying decision-making. We used rhesus macaques as our model organism. First we probed numerical decision-making, a form of abstract decision-making. We demonstrated that monkeys are able to compare discrete ratios, choosing an array with a greater ratio of positive to negative stimuli, even when this array does not have a greater absolute number of positive stimuli. Monkeys’ performance in this task adhered to Weber’s law, indicating that monkeys—like humans—treat proportions as analog magnitudes. Next we showed that monkeys’ ordinal decisions are influenced by spatial associations; when trained to select the fourth stimulus from the bottom in a vertical array, they subsequently selected the fourth stimulus from the left—and not from the right—in a horizontal array. In other words, they begin enumerating from one side of space and not the other, mirroring the human tendency to associate numbers with space. These and other studies confirmed that monkeys’ numerical decision-making follows similar patterns to that of humans, making them a good model for investigations of the neurobiological basis of numerical decision-making. </p><p>We sought to develop a system for exploring the neuronal basis of the cognitive and behavioral effects observed following transcranial magnetic stimulation, a relatively new, non-invasive method of brain stimulation that may be used to treat clinical disorders. We completed a set of pilot studies applying offline low-frequency repetitive transcranial magnetic stimulation to the macaque posterior parietal cortex, which has been implicated in numerical processing, while subjects performed a numerical comparison and control color comparison task, and while electrophysiological activity was recorded from the stimulated region of cortex. We found tentative evidence in one paradigm that stimulation did selectively impair performance in the number task, causally implicating the posterior parietal cortex in numerical decisions. In another paradigm, however, we manipulated the subject’s reaching behavior but not her number or color comparison performance. We also found that stimulation produced variable changes in neuronal firing and local field potentials. Together these findings lay the groundwork for detailed investigations into how different parameters of transcranial magnetic stimulation can interact with cortical architecture to produce various cognitive and behavioral changes.</p><p>Finally, we explored how monkeys decide how to behave in competitive social interactions. In a zero-sum computer game in which two monkeys played as a shooter or a goalie during a hockey-like “penalty shot” scenario, we found that shooters developed complex movement trajectories so as to conceal their intentions from the goalies. Additionally, we found that neurons in the dorsolateral and dorsomedial prefrontal cortex played a role in generating this “deceptive” behavior. We conclude that these regions of prefrontal cortex form part of a circuit that guides decisions to make an individual less predictable to an opponent.</p> / Dissertation

The possibility that nocturnal migrants use sunset as an orientational cue was explored in the White-throated Sparrow (Zonotrichia albicollis) between 1 April and 24 April, 1981. Orientation tests were run on the roof of the biology building at Western Kentucky University when birds were assumed to be in the proper physiological condition for migration. Birds exposed to only nighttime skies, birds isolated from all visual cues, and birds exposed to both daytime and nighttime skies did not exhibit the northward directional preference appropriate for the season tested. However, White-throated Sparrows exposed only to sunset and tested in the absence of visual nighttime cues exhibited significant orientation northward. These data clearly support the concept that White-throated Sparrows can use sunset as an orientational cue.

Ground meal and 4 sizes of pelleted feeds (3/16, ¼, 3/8 and ½ inch diameters) were offered to captive Starlings (Sturnus vulgaris, Linnaeus) to determine the minimum feed size they were unable to consume in significant amounts. The ground meal, 3/8 and ½ inch diameter pellets substantially decreased feed consumption compared with 3/16 and ¼ inch diameter pellets. Additionally, ½ inch pellets were consumed substantially less than ground meal or 3/8 inch pellets. Supplemental feed was require to prevent mortality among Starlings offered only ½ inch pellets. Two pellet sizes, the 3/16 inch pellet which was readily consumed by Starlings and the 3/8 inch pellet which was less preferred by the birds, were selected for tests with free ranging Starlings. Field tests comparing consumption of these 2 pellet sizes were conducted in southcentral Kentucky during January and February 1982. Starlings consumed significantly (P<0.01) lesser amounts of 3/8 inch pellets than 3/16 inch pellets during this test. It appears that feeding both ground meal and 3/8 inch diameter pelleted feeds to livestock is effective in reducing livestock feed losses to depredating Starling, and feeding larger-sized feeds such as ½ inch diameter pellets may further reduce losses.

The purpose of this research is to investigate the dispersion of ammonia (NH3) from three Concentrated Animal Feeding Operations (CAFOs) in western Kentucky, as well as to investigate the Weather Research and Forecasting – Chemistry (WRF-Chem) model’s sensitivity response to initial NH3 concentrations under both wet conditions (significant precipitation) and dry conditions (no precipitation). As expected, pollutant concentrations generally were significantly higher near their points of origin and generally declined away from the sources. Contrary to expectations, ammonia tended to rise through the planetary boundary layer (PBL) regardless of atmospheric conditions. Results showed modeled NH3 pollution levels at the surface generally to be higher under wet conditions. A GIS-based analysis method was developed to investigate model sensitivity to initial NH3 concentrations. Using this method, it was found that WRF-Chem exhibits an exponential relation between initial NH3 concentration and the final amount of NH3 produced by the model.