No abstract

Antioxidants could improve sperm media, extending the viability of spermatozoa and protecting their DNA. The protective ability of lipoic acid, melatonin, Trolox and crocin was tested on red deer spermatozoa incubated at 37°C. Cryopreserved spermatozoa were thawed and incubated with 1 mM or 0.1 mM of each antioxidant, with or without oxidative stress (100 μM Fe2+). Motility (CASA), viability, mitochondrial membrane potential and acrosomal status were assessed. Lipoperoxidation (malondialdehyde production), intracellular reactive oxygen species (ROS) and DNA status (TUNEL) were checked at 4 h. Incubation alone increased ROS and decreased motility. Oxidative stress intensified these effects, increasing lipoperoxidation and DNA damage. Lipoic acid had little protective effect, whereas 1 mM melatonin showed limited protection. Trolox lowered ROS and lipoperoxidation both in oxidised and non-oxidised samples. In oxidised samples, Trolox prevented DNA and acrosomal damage, and ameliorated motility. Crocin at 1 mM showed similar results to Trolox, but noticeably stimulated motility and had no effect on lipoperoxidation. In a second experiment, a broader range of crocin and melatonin concentrations were tested, confirming the effects of crocin (positive effects noticeable at 0.5–0.75 mM), but showing an increase in lipoperoxidation at 2 mM. Melatonin was increasingly effective at 2.5 and 5 mM (ROS, lipoperoxidation and DNA status). Crocin seems a promising new antioxidant, but its particular effects on sperm physiology must be further studied, especially the consequences of motility stimulation and confirming its effect on lipoperoxidation. Melatonin might be useful at relatively high concentrations, compared to Trolox.

Data from milk recording of Holstein-Friesian cows together with weather information from 2 regions in Southern Spain were used to define the models that can better describe heat stress response for production traits and somatic cell score (SCS). Two sets of analyses were performed, one aimed at defining the population phenotypic response and the other at studying the genetic components. The first involved 2,514,762 test-day records from up to 5 lactations of 128,112 cows. Two models, one fitting a comfort threshold for temperature and a slope of decay after the threshold, and the other a cubic Legendre polynomial (LP) model were tested. Average (TAVE) and maximum daily temperatures were alternatively considered as covariates. The LP model using TAVE as covariate showed the best goodness of fit for all traits. Estimated rates of decay from this model for production at 25 and 34°C were 36 and 170, 3.8 and 3.0, and 3.9 and 8.2g/d per degree Celsius for milk, fat, and protein yield, respectively. In the second set of analyses, a sample of 280,958 test-day records from first lactations of 29,114 cows was used. Random regression models including quadratic or cubic LP regressions (TEM_) on TAVE or a fixed threshold and an unknown slope (DUMMY), including or not cubic regressions on days in milk (DIM3_), were tested. For milk and SCS, the best models were the DIM3_ models. In contrast, for fat and protein yield, the best model was TEM3. The DIM3DUMMY models showed similar performance to DIM3TEM3. The estimated genetic correlations between the same trait under cold and hot temperatures (ρ) indicated the existence of a large genotype by environment interaction for fat (ρ=0.53 for model TEM3) and protein yield (ρ around 0.6 for DIM3TEM3) and for SCS (ρ=0.64 for model DIM3TEM3), and a small genotype by environment interaction for milk (ρ over 0.8). The eigendecomposition of the additive genetic covariance matrix from model TEM3 showed the existence of a dominant component, a constant term that is not affected by temperature, representing from 64% of the variation for SCS to 91% of the variation for milk. The second component, showing a flat pattern at intermediate temperatures and increasing or decreasing slopes for the extremes, gathered 15, 11, and 24% of the variation for fat and protein yield and SCS, respectively. This component could be further evaluated as a selection criterion for heat tolerance independently of the production level.

The aim of this study was to evaluate the effect of semen collection method (artificial vagina compared to electroejaculation), season in which the semen was collected (breeding season compared to non-breeding season), freezing extender (Biladyl(®), Andromed(®) and skim milk based extender) and pre-treatment procedure (washing compared to non-washing) on post-thaw semen quality in buck. Ejaculates from seven bucks of the Blanca-Celtibérica breed were collected by artificial vagina and electroejaculation during the breeding (July to December) and non-breeding season (January to June). Samples were split in two aliquots and one of them was washed. Three freezing extenders were evaluated on washing and non-washing sperm samples. Ejaculates collected by artificial vagina had a greater sperm quality after thawing, with greater values (P≤0.05) for SM (sperm motility), NAR (acrosome intact), YO-PRO-1-/PI- (intact spermatozoa), and Mitotracker+/YO-PRO-1- (spermatozoa with active mitochondria) and lower % DFI (DNA fragmentation index). Thawed sperm samples which were collected during the breeding season had greater values (P≤0.05) for NAR, intact spermatozoa and spermatozoa with active mitochondria, than those semen samples obtained during the non-breeding season. Semen freezing with Biladyl(®) and Andromed(®) resulted in a greater sperm quality (P≤0.05) after thawing in relation to milk-based extender. Washing procedure had no effect on sperm parameters assessed at thawing. Results from the present study suggest that the success of semen cryopreservation in Blanca-Celtibérica goat depends on semen collection method and season, as well as on the extender used. Thus, the post-thaw sperm quality will be greater (P≤0.05) when samples are collected by artificial vagina during the breeding season and when Biladyl(®) or Andromed(®) are used as freezing extenders.

Sperm design and velocity play key roles in influencing sperm performance and, therefore, can determine fertilization success. Several interspecific studies have demonstrated how these features correlate, and it has been hypothesized that selection may drive changes in these sperm traits. Here, we examine the association between sperm design and swimming velocity in a study conducted at an intraspecific level in Iberian red deer (Cervus elaphus hispanicus). We addressed how the structure of different sperm subpopulations, based on sperm morphometry and velocity, are interrelated and, in turn, how they associate with fertility. Our results show that males with high fertility rates have ejaculates with high percentages of spermatozoa exhibiting fast and linear movements and that these are highly correlated with a large proportion of spermatozoa having small and elongated heads. On the other hand, males with low fertility are characterized by a subpopulation structure in which slow and nonlinear as well as small and wide spermatozoa are predominant. These findings provide insight regarding how sperm size and velocity are interrelated and how they both are associated with fertility.