A Live-Attenuated Equine Influenza Vaccine Stimulates Innate Immunity in Equine Respiratory Epithelial Cell Cultures That Could Provide Protection From Equine Herpesvirus 1.

Equine herpesvirus 1 (EHV-1) ubiquitously infects horses worldwide and causes respiratory disease, abortion, and equine herpesvirus myeloencephalopathy. Protection against EHV-1 disease is elusive due to establishment of latency and immune-modulatory features of the virus. These include the modulation of interferons, cytokines, chemokines, antigen presentation, and cellular immunity. Because the modulation of immunity likely occurs at the site of first infection-the respiratory epithelium, we hypothesized that the mucosal influenza vaccine Flu Avert® I.N. (Flu Avert), which is known to stimulate strong antiviral responses, will enhance antiviral innate immunity, and that these responses would also provide protection from EHV-1 infection. To test our hypothesis, primary equine respiratory epithelial cells (ERECs) were treated with Flu Avert, and innate immunity was evaluated for 10 days following treatment. The timing of Flu Avert treatment was also evaluated for optimal effectiveness to reduce EHV-1 replication by modulating early immune responses to EHV-1. The induction of interferons, cytokine and chemokine mRNA expression, and protein secretion was evaluated by high-throughput qPCR and multiplex protein analysis. Intracellular and extracellular EHV-1 titers were determined by qPCR. Flu Avert treatment resulted in the modulation of IL-8, CCL2, and CXCL9 starting at days 5 and 6 post-treatment. Coinciding with the timing of optimal chemokine induction, our data also suggested the same timing for reduction of EHV-1 replication. In combination, our results suggest that Flu Avert may be effective at counteracting some of the immune-modulatory properties of EHV-1 at the airway epithelium and the peak for this response occurs 5-8 days post-Flu Avert treatment. Future in vivo studies are needed to investigate Flu Avert as a prophylactic in situations where EHV-1 exposure may occur.

Evaluation of systemic relaxin blood profiles in horses as a means of assessing placental function in high-risk pregnancies and responsiveness to therapeutic strategies.

Placental insufficiency is regarded as the primary factor contributing to late-term abortion and perinatal death of foals. Often when problems associated with late-term pregnancy in the horse are manifest the condition is well-advanced and therapeutic intervention may not be effective in rescuing the pregnancy. If a compromised pregnancy due to placental insufficiency could be identified early, the pregnancy might be sustained through medical intervention. Because the placenta is the sole source of circulating relaxin in the mare, we hypothesized that systemic relaxin may serve as a biomarker of placental function and fetal well-being and a predictor of pregnancy outcome at delivery. To test this hypothesis we monitored plasma relaxin in mares (light breeds) with normal and problematic pregnancies from clinical cases presented to the veterinary hospital and in pregnant mares experimentally inoculated with Streptococcus equi zooepidemicus to induce uterine infection. Upon establishment of placentitis, mares were assigned to different therapeutic strategies and responsiveness was monitored. Blood was collected during the third trimester of pregnancy, and relaxin content was determined using a homologous equine relaxin radioimmunoassay. The results reported here show a positive relationship between low circulating relaxin and poor pregnancy outcome in mares with compromised placental function. While relaxin may have value as a diagnostic assay for identifying mares with high-risk pregnancies associated with placental dysfunction, the variable results obtained from mares undergoing drug treatment for experimentally induced placentitis make it difficult to determine the reliability of relaxin for evaluating therapeutic efficacy.

Ultrasonographic assessment of fetal well-being during late gestation: development of an equine biophysical profile.

Mares with complicated pregnancies (illness, problems at parturition or delivery of an abnormal foal, n = 30) were scanned transabdominally from 298 days gestation to term in order to measure fetal size, evaluate fetal well-being and characterise the intrauterine environment. The results of the last scan obtained prior to parturition were compared to normal data obtained from fetuses of comparable gestational age to develop a biophysical profile specific for the equine fetus. Twelve mares produced a normal foal (positive outcome) and 18 mares delivered 19 abnormal foals (negative outcome). Both fetuses that were inactive throughout the entire scan and 4 of 5 fetuses with heart rate abnormalities were abnormal at birth. Three of 4 fetuses surrounded by decreased allantoic fluid quantities had a negative outcome. All mares with large anechoic spaces between the uterus and placenta (n = 3) and/or thickened uteroplacental units (n = 5) delivered abnormal foals. There was a significant correlation between fetal aortic diameter and neonatal foal weight in these complicated pregnancies (P<0.0001, r = 0.85). Fetal aortic diameters were predicted from maternal weight and 6 fetuses had smaller than predicted aortic diameters, all with negative outcomes. A biophysical profile of the equine fetus from 298 days gestational age to term was developed that included 6 factors related to pregnancy outcome: fetal heart rate, fetal aortic diameter, maximal fetal fluid depths, uteroplacental contact, uteroplacental thickness and fetal activity. The profile proved informative about fetal well-being, perinatal morbidity and perinatal mortality. A low score was a definite indication of an impending negative outcome; however, a high score was not assurance of a positive outcome. The utility of such a biophysical profile and future directions for research are discussed.