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Repeatable methods for IVF have not been established in the horse, reflecting the failure of standard capacitating media to induce changes required for fertilization capacity in equine sperm. One important step in capacitation is membrane cholesterol efflux, which in other species is triggered by cholesterol oxidation and is typically enhanced using albumin as a sterol acceptor. We incubated equine sperm in the presence of calcium, BSA, and bicarbonate, alone or in combination. Bicarbonate induced an increase in reactive oxygen species (ROS) that was abolished by the addition of calcium or BSA. Bicarbonate induced protein tyrosine phosphorylation (PY), even in the presence of calcium or BSA. Incubation at high pH enhanced PY but did not increase ROS production. Notably, no combination of these factors was associated with significant cholesterol efflux, as assessed by fluorescent quantitative cholesterol assay and confirmed by filipin staining. By contrast, sperm treated with methyl-b-cyclodextrin showed a significant reduction in cholesterol levels, but no significant increase in PY or ROS. Presence of BSA increased sperm binding to bovine zonae pellucidae in all three stallions. These results show that presence of serum albumin is not associated with a reduction in membrane cholesterol levels in equine sperm, highlighting the failure of equine sperm to exhibit core capacitation-related changes in a standard capacitating medium. These data indicate an atypical relationship among cholesterol efflux, ROS production, and PY in equine sperm. Our findings may help to elucidate factors affecting failure of equine IVF under standard conditions.

Assisted reproductive technologies (ART) revolutionised human fertility by providing treatment to patients previously considered incapable for reproductive purposes (Henkel & Schill, 2003). In 2017, it was estimated that approximately 6 million infants were born through ART since 1978, when the first baby produced by in vitro fertilisation (IVF) was born. It has been reported that about half of these births were in the past decade, highlighting the increasing use of these technologies, (Hunter, 2017). Initially, ART research was mainly focused on comprehending the factors that increase the likelihood of a successful pregnancy and

To establish a successful strategy for managing wild rabbit (Oryctolagus cuniculus algirus) populations it is essential to have a clear understanding of reproductive biology. In Portugal, previous work suggested a seasonal pattern of reproductive activity for this species. In this study we present additional information on the seasonal reproductive activity of the wild rabbit as well as data on the influence that environmental factors have on the onset and length of the breeding season. The study was carried out in Pancas, southern Portugal, from October 1997 to September 1998. Rabbits were collected every two months and post mortem analyses were performed. In the males, circulating concentration of testosterone, gonadal weight, daily sperm production, diameter of the seminiferous tubules and the thickness of epithelial cells were measured. The numbers of corpora lutea, embryos and placental scars were recorded in the females. To assess abundance and quality of food, samples of herbaceous vegetation were collected and analysed for water, fibre and crude protein content. The reproductive season extended from November to June with a peak in March/April, with both sexes showing an annual cycle of gonadal development and subsequent regression. All analysed females were pregnant and/or lactating between March and June. The mean litter size estimated by counting the embryos in the uteri was 3.90 ± 0.50. Daily sperm production was highest between November and April. Testicular function was strongly correlated with environmental temperature and the water content of the vegetation, and both testicular and ovarian development were correlated with vegetation biomass.

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The murine model has been widely used to study the host immune response to Neospora caninum. However, in most studies, the intraperitoneal route was preferentially used to establish infection. Here, C57BL/6 mice were infected with N. caninum tachyzoites by the intragastric route, as it more closely resembles the natural route of infection through the gastrointestinal tract. The elicited T-cell mediated immune response was evaluated in the intestinal epithelium and mesenteric lymph nodes (MLN). Early upon the parasitic challenge, IL-12 production by conventional and plasmacytoid dendritic cells was increased in MLN. Accordingly, increased proportions and numbers of TCRαβ+CD8+IFN-γ+ lymphocytes were detected, not only in the intestinal epithelium and MLN, but also in the spleen of the infected mice. In this organ, IFN-γ-producing TCRαβ+CD4+ T cells were also found to increase in the infected mice, however later than CD8+ T cells. Interestingly, splenic and MLN CD4+CD25+ T cells sorted from infected mice presented a suppressive activity on in vitro T cell proliferation and cytokine production above that of control counterparts. These results altogether indicate that, by producing IFN-γ, TCRαβ+CD8+ cells contribute for local and systemic host protection in the earliest days upon infection established through the gastrointestinal tract. Nevertheless, they also provide substantial evidence for a parasite-driven reinforcement of T regulatory cell function which may contribute for parasite persistence in the host and might represent an additional barrier to overcome towards effective vaccination.