Abstract: Scotopic vision in horses (Equus caballus) was investigated using behavioral measurements for the first time. Four horses were tested for the ability to make simple visual discriminations of geometric figures (circles and triangles) under various brightness levels within an enclosed building. Measurements of brightness ranging from 10.37 to 24.12 magnitudes per square arcsecond (mag/arcsec2; in candelas per square meter--7.70 to 2.43E-05 cd/m2) were taken using a Sky Quality Meter. These values approximated outdoor conditions ranging from twilight in open country to a dark moonless night in dense forest. The horses were able to solve the discrimination problems in all brightness settings up to 23.77 mag/arcsec2 (3.35E-05 cd/m2). Moreover, they easily navigated their way around obstacles located within the testing area in extremely dim light (>23.50 mag/arcsec2; 4.30E-05 cd/m2), which were in conditions too dark for the human experimenters to see. These findings support physiological data that reveal a rod-dominated visual system as well as observations of equine activity at night.

Abstract: The aggregation capacity of human erythrocytes lies between that of the non-aggregating bovine erythrocytes and the remarkably aggregating equine ones. As the ability to aggregate is attributed to cell factors and the composition of the plasma proteins, the role that plasma proteins play in the aggregation process in these three species was studied. Washed erythrocytes were suspended in phosphate-buffered saline (PBS; pH 7.4, 300 mOsm/L) plus polyvinylpyrrolidone (PVP) in a suitable concentration to obtain an average intensity of aggregation (control media). The superimposed effect of replacing 80% of the medium by either autologous plasma, serum or albumin solution was studied. The plasma proteins appeared to enhance aggregation by human and equine erythrocytes, but impaired this process in bovine erythrocytes. Some evidence was obtained supporting the existence of serum factors capable of reducing aggregation of erythrocytes in cattle and it was concluded that the non-aggregating behaviour of bovine erythrocytes may be due to the cells interacting particularly with the macromolecules in the serum.

Abstract: The aim of this study was to evaluate the behavior and physiologic responses of mares to removal from an established pasture herd and to isolation in a pasture setting for 6 h (Group I, n=5). Responses of mares in Group I were compared to mares that were transported and returned to the herd (Group T, n=5) and to mares moved to the isolation pasture with a companion (Group C, n=5). Behavior was recorded continuously for 6 h on the day before the isolation procedures (baseline, Day 0) and again on the day of the procedure (test, Day 1). Plasma cortisol, white blood cell count (WBC), neutrophil:lymphocyte ratio (N:L), and hematocrit (HCT) were measured once on Day 0 (a.m.) and twice on Day 1 (a.m. and p.m.). Heart rate (HR) was monitored continuously during Day 0 and Day 1. Intradermal response to phytohemagglutinin (PHA) injection was measured 18 h following injection, which was administered at the end of Day 1. Average time spent standing alert increased (P<0.05) in Groups I and C and average time spent grazing decreased (P<0.05) in Group C from Day 0 to Day 1. Also, there was a significant difference between groups (based on a calculated χ2-square value) in the proportion of mares that autogroomed, defecated, urinated, rolled, and whinnied on Day 1. Activity shift rate (ASR) and temperament scores increased significantly in Groups I and C from Day 0 to Day 1 (P<0.05). Plasma cortisol increased significantly in all groups from Day 0 to Day 1, a.m. (P<0.05) and decreased significantly from Day 1, a.m. to Day 1, p.m. (P<0.05). HCT significantly increased in all three groups from Day 0 to Day 1, a.m. (P<0.05). WBC significantly increased in Group T from Day 0 to Day 1, a.m. (P<0.05). N:L ratio significantly increased in Groups I and C from Day 0 and Day 1, a.m. to Day 1, p.m. (P<0.05). A variety of measures did indicate a response to removal from the pasture group, however, the overall, short-term response was minimal. Since the responses of Groups I and C were similar, the effects of isolation versus a novel environment or separation from the established herd could not be differentiated.

Abstract: The equilibrium unfolding and the kinetics of unfolding and refolding of equine lysozyme, a Ca2+-binding protein, were studied by means of circular dichroism spectra in the far and near-ultraviolet regions. The transition curves of the guanidine hydrochloride-induced unfolding measured at 230 nm and 292.5 nm, and for the apo and holo forms of the protein have shown that the unfolding is well represented by a three-state mechanism in which the molten globule state is populated as a stable intermediate. The molten globule state of this protein is more stable and more native-like than that of α-lactalbumin, a homologous protein of equine lysozyme. The kinetic unfolding and refolding of the protein were induced by concentration jumps of the denaturant and measured by stopped-flow circular dichroism. The observed unfolding and refolding curves both agreed well with a single-exponential function. However, in the kinetic refolding reactions below 3 M guanidine hydrochloride, a burst-phase change in the circular dichroism was present, and the burst-phase intermediate in the kinetic refolding is shown to be identical with the molten globule state observed in the equilibrium unfolding. Under a strongly native condition, virtually all the molecules of equine lysozyme transform the structure from the unfolded state into the molten globule, and the subsequent refolding takes place from the molten globule state. The transition state of folding, which may exist between the molten globule and the native states, was characterized by investigating the guanidine hydrochloride concentration-dependence of the rate constants of refolding and unfolding. More than 80% of the hydrophobic surface of the protein is buried in the transition state, so that it is much closer to the native state than to the molten globule in which only 36% of the surface is buried in the interior of the molecule. It is concluded that all the present results are best explained by a sequential model of protein folding, in which the molten globule state is an obligatory folding intermediate on the pathway of folding.

Abstract: The use of molecular techniques has increased our understanding of biological systems; in particular how genes and proteins operate within a cell or organ in response to various stimuli. These techniques can be used to study modes of action or mechanisms in disease states, or in response to applied management or nutritional changes. While applied and basic research are often conducted independent of one another, the real advantages to the equine industry will come from the use of molecular techniques to address applied challenges and/or opportunities within the industry. Therefore, the overall goal of our research has been to use basic and molecular tools to investigate modes of action resulting from changes in nutrition or management. We investigated active nutrient transport capacities of the small and large intestine, the effects of feed withdrawal on intestinal transporter gene expression and function, as well as other measurements pertaining to gut health (gut barrier function and gastrointestinal morphology). This novel and thorough approach to characterize sectional differences (fore- and hind- gut) of nutrient absorption has resulted in quantitative measures of active transport capacity for glucose, phosphorus, glutamine, lysine, and di-peptides in the horse. This research demonstrates that by-products of fermentation in the cecum such as microbial proteins and hydrolyzed phytate phosphorus, can be absorbed in the colon. Previously unidentified in the hindgut of the horse, we reported the expression of the di-and tri-peptide transporter, PepT1, as well as the neutral amino acid transporter, ASCT2, and transport of their substrates. We observed that nutrient transporters for glucose (SGLT1), glutamine (ASCT2) and for di-peptides (PepT1) can be regulated by diet, implying biological relevance to feeding management strategies. Feed withdrawal in horses pre- or post-surgery, prior to or during horse shows, during transport, or for other management reason is common. It is therefore important to understand how the removals of nutrients impacts gut health and/or disease susceptibility. The use of molecular techniques has provided a basis for the roles of certain core nutrients during times of feed withdrawal. For example, nutrients such as glutamine play a major role in proliferation and repair in the gut, serve as a major energy substrate, and may provide a means of assessing nutritional stress in the gut. Recent data concludes that following a feed withdrawal, the need for glutamine increases up to 116% in the proximal jejunum, and the gene expression of its transporter (ASCT2) increases by 78%. Additional studies utilizing molecular techniques to address nutritional challenges in the horse include how hormone loss due to ovariectomization impacts the homeostatic regulation of calcium and phosphorus in the horse. To improve nutrition and feeding management practices in horses, knowledge regarding how and where nutrients are absorbed, utilized and regulated under varying physiologic stressors (disease, exercise, reproductive status, etc.) is needed. The integration of applied and basic research facilitates a better and more complete understanding of mechanisms underlying common health and nutritional challenges facing the equine industry.