Updates and Current Challenges in Reproductive Microbiome: A Comparative Analysis between Cows and Women.

The microbiota plays an important role in numerous physiological processes, pathogenesis, development, and metabolism in different animal species. In humans, several studies have demonstrated an association between the vaginal microbiota and fertility rates, and even success in assisted reproduction techniques. In the context of cattle reproduction, although few studies have addressed the microbiota in a healthy state (which is not associated with diseases that affect the reproductive tract of cows), changes in its composition also seem to influence fertility. This review aims to explain the importance of the reproductive microbiota in female bovines and what is available in the literature regarding its possible role in increasing fertility. What are the challenges involved in this process? Future perspectives on its use and manipulation as a selection or intervention tool. Will it be possible to one day extrapolate the findings to reality and apply them in the field? In short, understanding the role of the reproductive microbiota of female bovines can signal the prospect of increasing production, whether of milk or meat, from the same number of animals, as it can optimize reproductive efficiency and perhaps become an allied tool for the economic profitability and sustainability of livestock farming.

Investigation of the vaginal microbiota of dairy cows through genetic sequencing of short (Illumina) and long (PacBio) reads and associations with gestational status.

The vaginal microbiota has been shown to be important in local immune regulation and may play a role in reproduction and fertility. Next-generation sequencing (NGS) technologies have been used to characterize the bovine vaginal microbiota, mainly using short-read sequencing (Illumina). However, the main limitation of this technique is its inability to classify bacteria at the species level. The objective of this study was to characterize the bovine vaginal microbiota at the species level using long-read sequencing (PacBio) and to compare it with the results of short-read sequencing. In addition, the vaginal microbiota of cows that became pregnant after artificial insemination (AI) was compared with that of infertile animals. Thirteen Holstein cows had vaginal swabs collected prior to AI. DNA was extracted and subjected to Illumina and PacBio sequencing to characterize the V4 region and the entire 16S rRNA gene, respectively. PacBio sequencing yielded 366,509 reads that were assigned to 476 species from 27 phyla. However, none of the most abundant reads (>1%) could be classified at the species level. Illumina sequencing yielded more reads and consequently was able to detect a more observed species, but PacBio sequencing was able to detect more unique and rare species. The composition of the vaginal microbiota varies according to the sequencing method used, which might complicate the interpretation of results obtained in the majority of the current studies. The present study expands on the current knowledge of bovine microbiota, highlighting the need for further efforts to improve the current databanks.

Correlation of testicular artery Doppler velocimetry with kinetics and morphologic characteristics of epididymal sperm in dogs.

Sperm quality can be affected by a reduction in testicular blood flow, which can be measured by Doppler ultrasonography. The aim of this study was to correlate the Doppler velocimetry of the testicular artery with kinetics of the epididymal spermatozoa in dogs. Twenty-two dogs (44 testicles) were evaluated by Doppler ultrasonography in five regions of the testicular artery before orchiectomy. Spermatozoa were recovered by the epididymal tail compression technique and analysed for kinetics on a computer-assisted semen analysis (CASA system). Morphology (modified Karras) and sperm membrane integrity were analysed by eosin-nigrosine staining. Data were analysed by Pearson's correlation test (p < .01). The mean total motility was 69.0% ± 17.7, progressive motility was 43.7% ± 14.7, average path velocity (VAP) was 127.0 µm/s ± 20.7, curvilinear velocity (VCL) was 221.0 µm/s ± 31.1, and sperm velocity index (SVI) was 389.9 ± 56.1. There were positive correlations between the peak systolic velocity (PSV) in the proximal supratesticular region with the SVI (r = .529), VCL (r = .555) and VAP (r = .473), and a negative correlation with the percentage of slow spermatozoa (r = -.463). The results suggest that the testicular artery blood flow velocity can positively affect the speed of spermatozoa movement. For the first time, we have correlated sperm kinetics with the Doppler evaluation of the testicular artery in dogs.

Can testicular artery Doppler velocimetry values change according to the measured region in dogs?

Testicular tissue maintenance is performed by the testicular artery, and the hemodynamics of this artery can be evaluated using Doppler ultrasonography. The aim of this study was to characterize the peak systolic velocity (PSV), end diastolic velocity (EDV), pulsatility index (PI) and resistivity index (RI) of five regions of the testicular artery in dogs, including two proposed regions and three that have been previously described. Twenty-two dogs were used, and the PSV, EDV, PI and RI of the testicular artery were measured in five regions: proximal, medial and distal supratesticular; marginal; and intratesticular. The median values for PSV (cm/s), EDV (cm/s), PI and RI in the five regions were as follows: proximal supratesticular (23.1, 3.7, 2.1 and 0.8); medial supratesticular (17.2, 4.5, 1.5, and 0.7); distal supratesticular (12.2, 5.7, 0.8, and 0.5); marginal (11.3, 6.5, 0.5, and 0.4); and intratesticular (5.7, 3.5, 0.5, and 0.4). There was a difference between the PSV of the medial and distal supratesticular regions. There were differences in the PSV, EDV, PI and RI among the distal supratesticular, marginal and intratesticular regions. Measurements of PSV, EDV, PI and RI of the testicular artery in dogs at the proposed regions showed different results due to the hemodynamic and morphological differences of the artery during its course in the spermatic cord and to the testicles. It is necessary to identify the region in testicular artery Doppler velocimetric evaluations of dogs, given that there is a difference according to the region measured.

Influence of cooling temperature in sperm subpopulations of domestic cats.

The objective of this study was to identify and compare domestic feline sperm subpopulations, chilled at -1 °C for 24 and 48 h, as well as to analyze the sperm frequency in different subpopulations. Ten adult cats were used. Sperm collection was performed using electroejaculation (EEJ). Spermatic kinetics were evaluated using a computerized system at three moments: fresh, 24 and 48 h after refrigeration. The ejaculates were divided into a group refrigerated at -1 °C (n = 5,) and a group refrigerated at 4 °C (n = 5),. A total of 1560 spermatozoa were analyzed individually, and the sperm subpopulations were identified using multivariate statistics. Three spermatic subpopulations were defined using prior analysis of the hierarchical dendrogram. A principal components analysis (PCA) identified the existence of three groups with higher iterations at the three moments: PC1 (VAP, VCL, VSL, ALH, SVI), PC2 (STR, LIN, WOB and SMI) and PC3 (BCF). Subpopulation 1, after 48 h of refrigeration at -1 °C, and subpopulation 3, after 24 h of refrigeration at 4 °C, maintained their sperm quality, which allowed us to characterize the groups of spermatozoa that were resistant to cryopreservation. The present study identified three well defined ejaculate spermatozoa subpopulations, with proportional distributions between the groups and two refrigeration resistant subpopulations.