Sperm parameters in the Great Dane: Influence of age on semen quality.

Not all sires have sperm suitable for chilled or frozen storage, and success in artificial insemination (AI) varies highly among individual dogs and breeds. Fertilizing potential is further complicated as sperm quality declines with the aging process. Due to the rapidity of aging and senescence in large breed dogs, associated health and fertility changes may be observed over a shorter period, though this period remains undefined for any breed. Working with a population of purebred Great Danes (GD), our aims were (1) to characterize the distribution of a series of sperm parameters, (2) to distinguish sources of variation in sperm quality within this rapidly aging breed, and (3) to identify changes in sperm quality that may accompany aging. Ejaculates collected from young, middle-aged, and senior Great Dane dogs (n = 50) were evaluated for semen volume, total sperm number and viability, and reactive oxygen species (ROS), in addition to sperm morphology and kinematic parameters. Total testicular volume was also determined using ultrasonography. Testicular volume was not a predictor of sperm production in the GD, however, significant differences between coat colors were identified. Age was negatively associated with total motility, progressive motility, and amplitude of lateral head displacement (ALH) (p < .05). We identified significant relationships between GD male age and TM, PM, and immotility with -9.9%, -9.0%, and +8.3% change per year of age, respectively, which support the anecdotal reports of decline of the fertility with the advance of age in this breed. Sperm of younger GD dogs aged 12 ≤ x < 24 months had significantly higher TM, PM, ALH, and nonlinear motility (p < .05) than older dogs (x ≥ 48 months). High ROS levels were positively associated with TM and PM, average pathway distance (DAP) and straight line distance (DSL), average pathway velocity (VAP), straight line velocity (VSL), and the presence of hairpin tails (p < .05). While age and ROS have significant influences on sperm parameters in the GD, the influence of selection for breed specific phenotypes could help explain the functional significance of the diversity among GD males.

Comparative oxidative metabolism in mammalian sperm.

In this review, we discuss the physiology of mitochondrial function in sperm using a comparative species approach. Mitochondria impart the ability for sperm from internal fertilizing species to attain individual motility and the ability to navigate the female reproductive tract to the site of fertilization. The presence of reactive oxygen species (ROS) is a normal physiological event of the mitochondrial electron transport chain (ETC); however, when excessive leakage of ROS occurs, sperm damage may follow. ROS production is associated with high levels of sperm motility but must be delicately balanced to prevent cellular damage during post-ejaculatory transport events. We discuss the differences in fundamental oxygen and ATP substrate balance in three mammalian species of veterinary importance, with an emphasis on ETC function, ROS production, and the balance of glycolytic and oxidative phosphorylation production of ATP in sperm.

Effects from disruption of mitochondrial electron transport chain function on bull sperm motility.

Sperm mitochondrial function is essential for normal physiology and fertility, but the importance of mitochondrial activity to support specific sperm functions, such as motility, varies between species. It was previously believed that mitochondrial function was not necessary for bull sperm motility [1]; however, this theory is contradicted by recently reported findings that the upper fraction of bull sperm swim-up preparations had both high motility and elevated mitochondrial oxygen consumption rates [2]. The objective of this study was to investigate the relationship between mitochondrial function and motility in bull sperm. We hypothesized that sperm motility would be positively correlated with mitochondrial oxygen consumption (MITOX) but unaffected by pharmacological inhibition of electron transport chain (ETC) activity. This was accomplished by monitoring both mitochondrial oxygen consumption and motility parameters in the presence of mitochondrial effector drug treatments. Duplicate ejaculates were collected by electroejaculation from Black Angus bulls (n = 4). Oxygen consumption, as % air saturation (pO2; oxygen partial pressure), over time was monitored in the presence of 5 drug treatments: vehicle control, FCCP, Antimycin (ANTI), Oligomycin (Oligo), and FCCP + Oligomycin (FCCP + OLIGO). Duplicate aliquots were prepared for concurrent motility assessment by computer-assisted sperm analysis (CASA) at 6 and 30 min post-treatment (t6 and t30). The impact of treatments on pO2 (in % air saturation) over time were assessed by generalized linear mixed effects modeling (GLMM) which was also used to test for differences in average motility across treatment conditions and time points (t6 and t30). Pearson product-moment correlation was used to investigate relationships between oxygen consumption and motility parameters. Overall, pO2 differed over time between treatment conditions (p < 0.0001). When compared to the vehicle treatment, ANTI and OLIGO significantly inhibited oxygen consumption (p < 0.05, adjusted), and FCCP stimulated a marked increase in oxygen consumption. No significant differences in motility over time were observed between treatments, so comparison of motility parameters between treatment conditions was performed with pooled timepoints. Motility parameters were only observed to differ significantly from the vehicle with ANTI Treatment, for which significant decreases in numerous parameters, including total motility (p = 0.007), progressive motility (p = 0.01), DAP (p = 0.01), VAP (p = 0.01) and VSL (p = 0.02) were identified. For the vehicle treatment, correlational analysis did not reveal any significant correlations between pO2 and any motility parameters at t6; however, several significant correlations were identified at t30. Mean pO2 was negatively correlated with local motility (p < 0.01) and positively correlated with DCL, DAP, and VCL (p < 0.05). Results from this study suggest that bovine sperm motility is impacted by mitochondrial functionality, with ETC inhibition by ANTI causing significant reduction in motility parameters. This study also demonstrates the use of a new technology for the assessment of bovine sperm mitochondrial function. This modality for evaluation of bull sperm mitochondrial function will inform future efforts to understand bull sperm function and fertility and aid investigations into toxicological agents.

Assessment of an iPad-based sperm motility analyzer for determination of canine sperm motility.

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 CVs 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.

Sperm mitochondrial regulation in motility and fertility in horses.

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.

Draft Genome Sequences of 1,183 Salmonella Strains from the 100K Pathogen Genome Project.

Salmonella is a common food-associated bacterium that has substantial impact on worldwide human health and the global economy. This is the public release of 1,183 Salmonella draft genome sequences as part of the 100K Pathogen Genome Project. These isolates represent global genomic diversity in the Salmonella genus.

100K Pathogen Genome Project: 306 Listeria Draft Genome Sequences for Food Safety and Public Health.

Listeria monocytogenes is a food-associated bacterium that is responsible for food-related illnesses worldwide. This is the initial public release of 306 L. monocytogenes genome sequences as part of the 100K Pathogen Genome Project. These isolates represent global genomic diversity in L. monocytogenes.