The early identification of infertile males improves reproduction efficiency at a production level and is essential to secure high production rates. Before entering a breeding program, males must pass an initial breeding soundness examination (BSE) which consist of several diagnostic exams whose end point is to estimate their future fertility. There is ongoing research to find new markers that allow better identification of fertile males. Doppler mode allows the quantitative and qualitative evaluation of the irrigation of organs. When evaluating the reproductive system, Doppler mode has been successfully used for the evaluation of the uterine and ovarian irrigation. In males, it is gaining relevance for the evaluation of testicular irrigation. Researchers have found a relationship between pulsed-wave Doppler velocimetric parameters and seminal quality in various domestic animal species. This suggests Doppler ultrasound parameters should be considered as objective parameters to evaluate testicular function. In this review, we analyze the results in the main domestic animal species and discuss the differences and similarities among the results. We also discuss the effect of the location of the measurements, breed, season and laterality in the measurement of Doppler velocimetric parameters as well as the impact and limitations of this method of assessing breeding soundness.

Controlled ovarian stimulation is a necessary step in some assisted reproductive procedures allowing a higher collection of female gametes. However, consequences of this stimulation for the gamete or the offspring have been shown in several mammals. Most studies used comparisons between oocytes from different donors, which may contribute to different responses. In this work, we use the bovine model in which each animal serves as its own control. DNA methylation profiles were obtained by single-cell whole-genome bisulfite sequencing of oocytes from pre-ovulatory unstimulated follicles compared to oocytes from stimulated follicles. Results show that the global percentage of methylation was similar between groups, but the percentage of methylation was lower for non-stimulated oocytes in the imprinted genes APEG3, MEG3, and MEG9 and higher in TSSC4 when compared to stimulated oocytes. Differences were also found in CGI of imprinted genes: higher methylation was found among non-stimulated oocytes in MEST (PEG1), IGF2R, GNAS (SCG6), KvDMR1 ICR UMD, and IGF2. In another region around IGF2, the methylation percentage was lower for non-stimulated oocytes when compared to stimulated oocytes. Data drawn from this study might help to understand the molecular reasons for the appearance of certain syndromes in assisted reproductive technologies-derived offspring.

The evolution of some fetal growth indices and arterial blood flow parameters in the umbilical cord of the embryos and fetuses of primiparous pregnant goats of Murciano-Granadina breed were analyzed by ultrasonography. Weekly ultrasonographic sessions took place from 18- to 125-days post-breeding. Fetal measures were carried out by ultrasound B-mode. This mode was used to take a series of measurements in the embryo/fetus throughout pregnancy: crown-rump length (CRL, from 24-days post-mating -dpm- to 61 dpm), trunk diameter (TD, 24–34 dpm), biparietal diameter (BPD, 28–125 dpm) and eye orbit diameter (EOD, 75–125 dpm). Spectral Doppler was used to study blood flow from umbilical artery. Different blood flow parameters were obtained as follows: Arterial Pulse, Peak Systolic Velocity (PSV), End Diastolic Velocity (EDV), Mean Velocity (MV), Systolic velocity/Diastolic velocity Ratio (S/D), Pulsatility Index (PI) and Resistance Index (RI). In this study, the umbilical cord was first noticed between 32- and 35-days post-breeding. However, these umbilical arterial blood flow parameters were not conclusive (positive S/D ratios and RI < 1) until 65–80 days of pregnancy. The explanation to these results could be that vascular development related to umbilical arteries elasticity and diameter is not good enough in early pregnancy. Therefore, these vessels have already acquired their appropriate characteristics in order to allow blood flow parameters and Doppler index measures from only 2.5 months of pregnancy. This is the first time that a detailed study of fetal growth indices and umbilical artery flow rates in fetuses from Murciano-Granadina goats has been performed throughout virtually the entire duration of gestation. In conclusion, the evolution of the fetal growth indices in this breed has determined that the umbilical artery velocimetric parameters (PSV, EDV, MV) increase significantly and the AP, S/D, PI and RI indices decrease significantly throughout the analyzed pregnancy period.

Artificial insemination (AI) with frozen semen in goats still presents serious difficulties, especially in certain goat breeds, in spite of technological progress. The aim of this work is the in vitro study of seminal extenders adapted from those used on other species to evaluate the response of goat sperm to several homeostatic conditions in order to achieve optimal post-thaw semen quality. Three different extenders based on different activity principles were used: (1) extender according to the methodology proposed for pigs, (2) skimmed-milk-based extender according to the methodology proposed for goats in France, and (3) a new egg-yolk-based extender replacing membrane-protective surfactants with sodium dodecyl sulfate (SDS) and named by our team as extender “IMIDA”. The freezing guidelines were those proposed for the freezing of porcine semen. The results obtained show that the egg-yolk-based extenders have good parameters of sperm motility at thawing, studied objectively using the computer-assisted sperm analysis (CASA) system and also subjectively. In particular, in the sperm resistance test after five hours, the thawed sperm containing SDS in their composition showed an optimal average on every evaluated parameter. The new IMIDA extender provided the highest sperm quality averages, so it could be a good extender to use in cryopreservation of semen in the caprine species.

The objective was to investigate the effects of sex-sorting on early fertilization events in ovum pick up (OPU) derived oocytes fertilized in vitro with frozen-thawed sperm at different co-incubation lengths. Eighty-four OPU sessions were carried out in 18 cyclic, dry and non-stimulated Holstein Friesian and German black pied cows. Ovum pick up oocytes were matured in vitro for 24 hours and fertilized with frozen-thawed sex sorted or unsorted sperm from the same ejaculate. Fertilization was achieved by two experimental protocols: 1) short gamete co-incubation length: 4, 8 and 12 hours; and 2) long gamete co-incubation length: 18 and 24 hours. After in vitro fertilization, ova were fixed and stained to identify early fertilization events. Sperm penetration, monospermy, pronuclear formation and syngamy did not differ, whether sexed or unsexed sperm was used. Overall, the findings demonstrate similar fertilizing potential between sex-sorted and unsorted sperm