Simon Paul De Graaf

Transitioning from theoretical learning to practical application remains a significant challenge for students in medical and veterinary science education, particularly in the context of medical imaging and ultrasound interpretation. Traditional lecture-based methods offer limited support for developing the spatial reasoning and technical skills required for ultrasound pregnancy diagnosis. This study evaluates the effectiveness of an interactive mixed reality (MR) training tool, Ewe Scan, delivered through the Apple Vision Pro, compared to traditional lecture-based instruction. Forty-two undergraduate students were randomly assigned to either a lecture-trained or MR-trained group and assessed immediately after training and again after six weeks. Results showed that MR-trained students significantly outperformed their lecture-trained peers in both immediate comprehension and retention over time, particularly in ultrasound interpretation skills. The MR-trained group also reported higher levels of engagement, confidence, and satisfaction with their training experience. These findings suggest that MR-based learning enhances educational outcomes by improving spatial understanding, increasing active engagement, and supporting knowledge retention. Integrating MR simulations into ultrasound education offers a scalable, ethical, and effective alternative to traditional training methods, contributing to advancements in medical imagery education.

Sperm DNA integrity is increasingly considered a useful measure of semen quality in mammalian reproduction. However, the definition of DNA integrity, the ideal means by which it should be measured, and its predictive value for fertility remain a topic of much discussion. With an emphasis on livestock species, this review discusses the assays that have been developed to measure DNA integrity as well as their correlation with in vitro and in vivo fertility.

This chapter on reproduction is concerned with various direct interventions to manipulate and enhance reproductive function in sheep flocks. Collectively referred to as controlled breeding, these interventions may encompass the limitation of reproduction and control of its timing; the increase in the number of progeny per ewe; artificial reproductive technologies such as artificial insemination (AI) and multiple ovulation and embryo transfer (MOET); and the induction of abortion or parturition. Synchronisation of oestrus, advancement of the breeding season, AI and MOET are of most interest to the Australian sheep industry, and as such will be the topics primarily discussed in this chapter.

For successful fertilisation to occur, spermatozoa need to successfully migrate through the female reproductive tract and penetrate the oocyte. Predictably, poor sperm motility has been associated with low rates of fertilisation in many mammalian species, including the ram. As such, motility is one of the most important parameters used for in vitro evaluation of ram sperm quality and function. This review aims to outline the mechanical and energetic processes which underpin sperm motility, describe changes in motility which occur as a result of differences in sperm structure and the surrounding microenvironment, and assess the effectiveness of the various methods used to assess sperm motility in rams. Methods of subjective motility estimation are convenient, inexpensive methods widely used in the livestock industries, however, the subjective nature of these methods can make them unreliable. Computer-assisted sperm analysis (CASA) technology accurately and objectively measures sperm motility via two-dimensional tracing of sperm head motion, making it a popular method for sperm quality assurance in domesticated animal production laboratories. Newly developed methods of motility assessment including flagellar tracing, three-dimensional sperm tracing, in vivo motility assessment, and molecular assays which quantify motility-associated biomarkers, enable analysis of a new range of sperm motion parameters with the potential to reveal new mechanistic insights and improve ram semen assessment. Experimental application of these technologies is required to fully understand their potential to improve semen quality assessment and prediction of reproductive success in ovine artificial breeding programs.

Semen preservation is an essential component of reproductive technologies, as it promotes genetic gain and long-distance semen transport and multiplies the number of ewes able to be inseminated per single ejaculate. However, the reduced temperature during cold storage at 5 or 15 °C inflicts sub-lethal damage to spermatozoa, compromising sperm quality and the success of artificial breeding. New and emerging research in various species has reported the advantages of storing spermatozoa at higher temperatures, such as 23 °C; however, this topic has not been thoroughly investigated for ram spermatozoa. Despite the success of storing spermatozoa at 23 °C, sperm quality can be compromised by the damaging effects of lipid peroxidation, more commonly when metabolism is left unaltered during 23 °C storage. Additionally, given the biosafety concern surrounding the international transport of egg-yolk-containing extenders, further investigation is critical to assess the preservation ability of synthetic extenders and whether pro-survival factors could be supplemented to maximise sperm survival during storage at 23 °C.

The preservation of rhinoceros semen is vital for captive breeding programs. While successful collection and cryopreservation of rhinoceros semen has been reported, the volume and quality of semen produced is often low due to the high viscosity associated with ejaculates collected via electroejaculation. Reducing semen viscosity would enable access to previously unusable spermatozoa from viscous fractions and could improve quality post-thaw. The enzyme papain successfully reduced the viscosity of camelid semen but has yet to be tested in wildlife species. This study assessed the influence of papain on the in vitro quality of rhinoceros spermatozoa during cryopreservation using advanced semen assessment. In experiment 1, the motility of spermatozoa from the viscous fraction of an ejaculate, either untreated or treated with papain and its inhibitor E-64 prior to cryopreservation, was assessed post-thaw. In experiment 2, spermatozoa from papain-treated viscous fractions were compared to spermatozoa frozen from untreated sperm-rich fractions pre-freeze, as well as after 0, 1.5 and 3 h of incubation post-thaw (37 °C). Papain significantly increased the quantity of spermatozoa collected from ejaculates, as well as the motility prior to freezing. Papain also improved the post-thaw motility, velocity, linearity and straightness of samples compared to sperm-rich samples, with no detriment to sperm viability, lipid membrane disorder, production of ROS or DNA integrity (p < 0.05). Results show the benefit of supplementing rhinoceros spermatozoa with papain prior to cryopreservation on sperm cryosurvival and demonstrates the potential of using papain to improve the success of cryopreservation protocols, not only for the rhinoceros, but also for other wildlife species.

This study assessed whether the seasonal effects of melatonin that upregulate ram reproductive function alter sperm global methylation or protamine deficiency and whether these parameters corresponded to ram endocrinology, semen production and quality. Ejaculates were assessed from rams that received melatonin implants (n = 9) or no implants (n = 9) during the non-breeding season. Ejaculates (n = 2/ram/week) were collected prior to implantation (week 0), 1, 6 and 12 weeks post implantation and during the following breeding season (week 30). Flow cytometry was used to assess the sperm global methylation and protamine deficiency in each ejaculate, which had known values for sperm concentration, motility, morphology, DNA fragmentation, seminal plasma levels of melatonin, anti-Mullerian hormone and inhibin A. Serum levels of testosterone and melatonin were also evaluated. Though there was no effect of melatonin or season, sperm protamine deficiency was negatively correlated with sperm production and seminal plasma levels of anti-Mullerian hormone and positively correlated with sperm DNA fragmentation and morphology. Global methylation of spermatozoa was positively correlated with sperm DNA fragmentation, morphology and serum testosterone and negatively correlated with sperm motility. These moderate associations with sperm production and quality suggest that sperm protamine deficiency and global methylation are indicative of ram testicular function.