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.

Abstract: Abstract The predominant housing system used for domestic horses is individual stabling; however, housing that limits social interaction and requires the horse to live in semi-isolation has been reported to be a concern for equine welfare. The aim of the current study was to compare behavioural and physiological responses of domestic horses in different types of housing design that provided varying levels of social contact. Horses (n = 16) were divided equally into four groups and exposed to each of four housing treatments for a period of five days per treatment in a randomized block design. The four housing treatments used were single housed no physical contact (SHNC), single housed semi-contact (SHSC), paired housed full contact (PHFC) and group housed full contact (GHFC). During each housing treatment, adrenal activity was recorded using non-invasive faecal corticosterone metabolite analysis (fGC). Thermal images of the eye were captured and eye temperature was assessed as a non-invasive measure of the stress response. Behavioural analysis of time budget was carried out and an ease of handling score was assigned to each horse in each treatment using video footage. SHNC horses had significantly higher (p = 0.01) concentrations of fGC and were significantly (p = 0.003) more difficult to handle compared to the other housing types. GHFC horses, although not significantly different, had numerically lower concentrations of fGC and were more compliant to handling when compared to all other housing treatments. Eye temperature was significantly (p = 0.0001) lower in the group housed treatment when compared to all other treatments. These results indicate that based on physiological and behavioural measures incorporating social contact into the housing design of domestic horses could improve the standard of domestic equine welfare.

Abstract: Three arthropod-borne alphaviruses, western equine encephalitis viruses (WEEV), eastern equine encephalitis viruses (EEEV) and Venezuelan equine encephalitis viruses are the aetiological agents of a sometimes severe encephalomyelitis in equines and humans in the New World. With regard to the different ecology and epidemiology of these viruses, a method applied in serological screening should be able to distinguish between them as well as other related members of the genus Alphavirus in the American continent. However, this has been hampered in the past by (a) the close antigenic relationship between alphaviruses in traditional serological assays, especially in the routinely used haemagglutination-inhibition, and (b) the need of biosafety level 3 facilities to grow the viral antigens. An epitope blocking assay using an EEEV glycoprotein E1-expressing recombinant Sindbis virus and virus-specific monoclonal antibodies (mAbs) binding to the E1 of EEEV (strain NJ/60) and the E1 of Sindbis virus was established using automated flow cytometry. The test was evaluated using sera of infected and vaccinated rabbits. A cut-off value of 30% inhibition for antigenic complex-specific seroconversion was found to be sufficient for the detection of the respective infection. By using three different mAbs in parallel, we were able to detect alphavirus genus-, EEEV- and WEEV-complex-specific serum antibodies. As this test is based on the inhibition of binding of virus-specific mAbs, sera of every origin other than mouse can be tested. Thus, this assay may prove useful in the serological screening of a variety of animal species during an outbreak investigation.

Abstract: Horses are likely to exhibit aggression when meeting for the first time. Therefore, this study compared 3 methods for mixing horses to evaluate their effectiveness in reducing aggressive interactions: (1) mixing pairs of horses in a paddock (P, 10 minutes, 15 tests), (2) introducing 1 unfamiliar horse to a pair of familiar, resident horses in a paddock (PP, 10 minutes, 15 tests), (3) allowing limited physical contact between pairs of horses for a short period of pre-exposure in neighboring boxes (B, 5 minutes, 16 tests) before mixing them in a paddock (BP, 10 minutes 16 tests). A total of 16 Swedish Standardbred mares, aged 6-18 years (mean age ± SD: 11 ± 4.4), were included in the study. Half of the horses were familiar with each other (resident horses, n = 8), whereas the other half were bought in from a variety of sources (unfamiliar horses, n = 8). Social interactions, consisting of behaviors from the sender, the receiver, and the subsequent sender's response, were recorded continuously as frequencies. There were no differences in the frequencies of aggressive behaviors between the 3 mixing methods, including those aggressive behaviors in which physical contact had been attempted (kick, strike). Although resident horses were overall more aggressive (median number of aggressive behaviors per horse, 62; Q1, 36; Q3, 68.5) than unfamiliar horses (median per horse, 4; Q1, 2; Q3, 12.5) during all tests (U = 97, P = 0.003), none of the 62 tests needed to be terminated. Unfamiliar horses did not receive more aggression from resident horses in PP (mean per test ± SD: 5.1 ± 3.1) than in P (mean per test ± SD: 6.4 ± 4.9) (t = 0.63, P = 0.544). However, the behavior “attack” was more frequent in PP (median per test, 2; Q1, 0; Q3, 5) than in P (median per test, 0; Q1, 0; Q3, 1) (U = 282, P = 0.042), and “flee” was more frequent in PP (median per test, 6; Q1, 4; Q3, 8) than in P (median per test, 1; Q1, 0; Q3, 6) (U = 290, P = 0.018). Pre-exposure in boxes did not reduce aggression in BP (median per test, 7; Q1, 4.3; Q3, 11.8) as compared with P (median per test, 6; Q1, 2; Q3, 16) (U = 264, P = 0.767), but during pre-exposure in B tests, horses exchanged more nonaggressive (median per test, 2; Q1, 0.3; Q3, 4) than aggressive (median frequency of aggressive behavior, 0; Q1, 0; Q3, 1) (W = 71, P = 0.013) and mixed interactions (median per test, 0; Q1, 0; Q3, 1) (W = 92, P = 0.016) through the opening. Results suggest mixing an unfamiliar horse with 2 resident horses at the same time instead of one by one may be preferable. In this way, the total aggression received by the unfamiliar horse will potentially be less, even though aggressive interactions may be more intense.