Mitchell Coyle

Ecdysterone controls moulting and reproduction in insects, crustaceans, and helminths. It is also produced by many plants, probably as an insect deterrent. The steroid is not made by vertebrates but has anabolic effects in mammals and could be useful for treating sarcopenia in aged horses. However, ecdysterone is banned in horseracing and equestrian sports, and with no limit of reporting, the risk of unintended exposure to this naturally occurring prohibited substance is a concern. To explore this risk, pasture plants and hay samples were analysed for ecdysterone content, as well as samples of blood, faeces, and intestinal mucosa from horses (euthanized for non-research purposes) with varying degrees of endo-parasite infestation. The variability in serum ecdysterone concentrations between different horses after administering a fixed dose was also examined. Ecdysterone was detected in 24 hay samples (0.09 to 3.74 µg/g) and several weeds, with particularly high concentrations in Chenopodium album (244 µg/g) and Solanum nigrum (233 µg/g). There was a positive correlation between faecal ecdysterone and faecal egg counts, but no effect of anthelmintic treatment and no relation to the number of encysted cyathostome larvae in the large intestine mucosa. Certain horses maintained an unusually high serum ecdysterone concentration over several weeks and/or displayed an abnormally large response to oral ecdysterone administration. Thus, the risk of environmental exposure to ecdysterone is apparent, and several factors must be considered when determining an appropriate dosage for clinical studies or setting a reporting threshold for equine sports.

The increased frequency of extreme weather events due to climate change has complicated the epidemiological pattern of mosquito-borne diseases, as the host and vector dynamics shift to adapt. However, little is known about the seroprevalence of common mosquito-borne virus infections in horses in Australia. In this study, serological surveys for multiple alphaviruses were performed on samples taken from 622 horses across two horse populations (racehorses and horses residing on The University of Queensland (UQ) campus) in Queensland using the gold standard virus neutralization test. As is the case in humans across Australia, Ross River virus (RRV) is the most common arbovirus infection in horses, followed by Barmah Forest virus, with an overall apparent seroprevalence of 48.6% (302/622) and 4.3% (26/607), respectively. Horses aged over 6 years old (OR 1.86, p = 0.01) and residing at UQ (OR 5.8, p < 0.001) were significantly associated with seroconversion to RRV. A significant medium correlation (r = 0.626, p < 0.001) between RRV and Getah virus (GETV) neutralizing antibody titers was identified. Collectively, these results advance the current epidemiological knowledge of arbovirus exposure in a susceptible host in Australia. The potential use of horses as sentinels for arbovirus monitoring should be considered. Furthermore, since GETV is currently exotic to Australia, antibodies cross-reactivity between RRV and GETV should be further investigated for cross-protection, which may also help to inform vaccine developments.

In 2011, following severe flooding in Eastern Australia, an unprecedented epidemic of equine encephalitis occurred in South-Eastern Australia, caused by Murray Valley encephalitis virus (MVEV) and a new variant strain of Kunjin virus, a subtype of West Nile virus (WNVKUN). This prompted us to assess whether a delta inulin-adjuvanted, inactivated cell culture-derived Japanese encephalitis virus (JEV) vaccine (JE-ADVAX™) could be used in horses, including pregnant mares and foals, to not only induce immunity to JEV, but also elicit cross-protective antibodies against MVEV and WNVKUN. Foals, 74–152 days old, received two injections of JE-ADVAX™. The vaccine was safe and well-tolerated and induced a strong JEV-neutralizing antibody response in all foals. MVEV and WNVKUN antibody cross-reactivity was seen in 33% and 42% of the immunized foals, respectively. JE-ADVAX™ was also safe and well-tolerated in pregnant mares and induced high JEV-neutralizing titers. The neutralizing activity was passively transferred to their foals via colostrum. Foals that acquired passive immunity to JEV via maternal antibodies then were immunized with JE-ADVAX™ at 36–83 days of age, showed evidence of maternal antibody interference with low peak antibody titers post-immunization when compared to immunized foals of JEV-naïve dams. Nevertheless, when given a single JE-ADVAX™ booster immunization as yearlings, these animals developed a rapid and robust JEV-neutralizing antibody response, indicating that they were successfully primed to JEV when immunized as foals, despite the presence of maternal antibodies. Overall, JE-ADVAX™ appears safe and well-tolerated in pregnant mares and young foals and induces protective levels of JEV neutralizing antibodies with partial cross-neutralization of MVEV and WNVKUN.