The biological nature of age‐related declines in fertility in males of any species, including stallions, has been elusive. In horses, the economic costs to the breeding industry are frequently extensive. Mitochondrial function in ejaculated sperm, which is essential for sperm motility, is reflected by adenosine triphosphate production, mitochondrial oxidative efficiency and production of reactive oxygen species, and that this balance may become compromised in ageing stallions and during the process of cryopreservation. This presentation will focus on mitochondrial integrity and function as an avenue for understanding the pathophysiology of sperm when undergoing cryopreservation and male ageing. We discuss the importance of understanding the differences and similarities of sperm mitochondria to that of somatic cells regarding structure and mitochondrial biochemistry relating to sperm function. The roles of oxidative phosphorylation and glycolysis in sperm mitochondria are outlined as is the method of determining oxygen consumption and calcium homoeostasis in sperm mitochondria. Further, we outline the role of oxidative stress and reactive oxygen species.

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The objective of this study was to evaluate the repeatability and accuracy of canine sperm motility (total and progressive) assessment with a tablet-based Canine iSperm ® instrument compared to computer-assisted sperm analysis (CASA). The experiment used fresh and frozen/thawed canine semen samples for comparisons of semen analysis parameters (concentration, total motility, and progressive motility) between a CASA system, iSperm ®, and NucleoCounter ® SP-100 ™ (concentration) instruments. Spearman’s Rho correlational analysis was used to identify significant associations between motility assessment methods. Significant positive correlations were found between CASA assessment and iSperm ® for both progressive and total motility measurements. We also determined the coefficient of variation (CV) for repeatability of sample analysis for iSperm ® and CASA for fresh sperm, wherein each sample was assessed 10 times on both devices. For fresh and frozen-thawed samples, concentration assessment by iSperm ® showed high variability (CV= 19.9 ± 1.5%). For iSperm ® assessment of total and progressive motility, the CV’s were 6.3 ± 0.5% and 10.7 ± 0.8%, respectively. The results indicate that the iSperm ® application offers an accurate and alternative measurement of motility to traditional CASA analysis, though caution should be taken when assessing concentration due to the high CV observed in this study.

OBJECTIVE To describe uterine prolapse, predispositions, and outcomes in mares treated between 1988 and 2019. ANIMALS 24 mares with uterine prolapse. PROCEDURES Clinical records were retrospectively reviewed, and follow-up information was gathered. The Mann-Whitney U test and Fisher exact test were used to analyze results for history and outcome variables for potential association with mare survival to hospital discharge. The χ2 test was used to compare breed distribution. RESULTS Age was known in 23 mares (median, 11.1 years). For 15 mares with exact known parity, the median parity was 3 births (range, 1 to 13 births). For 22 mares, the maiden status was known (7 [32%] maiden; 15 [68%] multiparous). Breed distribution differed significantly between mares with uterine prolapse and all mares treated at the Veterinary Medical Teaching Hospital. Arabians were overrepresented in the uterine prolapse group (7/24 [29%]), compared with all mares treated (4,174/44,474 [9%]). Uterine prolapse occurred within 2 hours after parturition for 10 of 11 mares with known foaling times, after dystocia for 7 mares, and after abortion for 4 mares. Seventeen of 23 (74%) mares survived to hospital discharge. Acute hemorrhage was the most frequent cause of death. There were no correlations between mare survival to hospital discharge and mare parity or age, foal sex, retained fetal membranes, sepsis, or maiden status. Colts were overrepresented in foals with a known sex (12/17). CLINICAL RELEVANCE Results indicated a breed predisposition to uterine prolapse, with Arabian mares overrepresented among affected mares. No characteristics correlated with mare survival to hospital discharge.

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Low levels of intracellular reactive oxygen species (ROS) are essential for normal sperm function and are produced by sperm mitochondria as a byproduct of metabolism, but in excess, ROS can cause catastrophic cellular damage and has been correlated with infertility, poor sperm motility and abnormal morphology in humans. Stallion sperm motility is fueled predominantly by oxidative phosphorylation‐produced ATP, requiring high basal rates of mitochondrial function. Consequently, whether elevated ROS production by stallion sperm is an indicator of dysfunctional or highly motile cells has been debated by researchers over the last decade. The objective of this study was to evaluate the relationship between various sperm morphologies and ROS production in fresh and cooled stallion semen by employing the novel method of imaging flow cytometry for stallion semen assessment. For evaluation of fresh semen, single ejaculates (n = 5) were collected from four resident stallions at the University of California, Davis. For the evaluation of 24‐h cool‐stored semen, single ejaculates were collected from stallions at Texas A&M University (n = 5) and shipped to the University of California, Davis overnight for evaluation. Ejaculate volume, sperm concentration and motility parameters were recorded. Samples were co‐stained for viability and ROS detection with SytoxGreen™ and dihydroethidium (DHE), respectively, and evaluated with the Amnis® ImageStream® system (Luminex Corporation). Antimycin, an electron transport chain inhibitor that triggers ROS production (1 μM), was used as a positive control for DHE, while dead cells (2× snap frozen in liquid nitrogen) served as a positive control for SytoxGreen™. Unstained samples were also evaluated as controls. Imaging flow cytometric analysis was performed with the ideas® software (Luminex Corporation). Evaluated morphologies included abnormal head (AH), abnormal midpiece (AM), abnormal tail (AT), proximal cytoplasmic droplet (PD), or distal cytoplasmic droplet (DD), and morphologically normal (MN) cells. For fresh semen, an additional abnormality, coiled tail and midpiece (CTM) was assessed; 24‐h cool‐stored semen did not contain enough viable CTM cells for analysis. Only cells with obvious, single abnormalities were selected for the first portion of analysis to minimize subjectivity. Mixed effects modelling was used to evaluate the relationship between each morphologic classification and the corresponding DHE fluorescence intensity. Compared to the MN population, ROS production was significantly higher in viable cells with AH, PD and AM (p < .0001) in both fresh and cooled semen. CTM cells had significantly higher levels of ROS production compared to MN cells in fresh semen (p < .0001). There was no significant difference in ROS levels between MN cells and AT and DD cells in either fresh or cooled semen (p > .05). These results suggest that ROS generation is indicative of abnormal cell morphology and function and confirm that imaging flow cytometry is a valuable tool for the asses...

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