Correlations of corpus luteum blood flow with fertility and progesterone in embryo recipient mares.

The aim of this study was to evaluate the correlation between the corpus luteum vascularization with the concentration of progesterone and the fertility of embryo recipient mares. Mangalarga Marchador mares (n = 33) were distributed into groups according to the days (D) after ovulation, as follows: D3 (n = 8), D4 (n = 8), D5 (n = 9), and D6 (n = 8). The evaluations of the corpus luteum, endometrium, and blood collection to quantify the progesterone concentration were carried out on D3, D4, D5, and D6. Among the parameters evaluated, only progesterone concentration on D6 differed from the other groups (P <0.05). A positive correlation (P <0.05) between the diameter and the area of the corpus luteum, and the objective and subjective methods of the corpus luteum vascular perfusion, was identified. Likewise, a positive correlation (P <0.05) was observed between the objective and subjective methods of the vascular perfusion in the corpus luteum and the progesterone concentration. The pregnancy rate obtained in this study (54.54%) was not affected (P> 0.05) by the day of embryo transfer, whose percentages were 37.50% (3/8) on D3, 50% (4/8) on D4, 66.70% (6/9) on D5, and 62.50% (5/8) on D6. It was estimated that with each increase on the day of embryo transfer, the pregnancy rate increases. The results allow to conclude that the corpus luteum vascularization in mares, evaluated by Doppler ultrasound, correlates with progesterone concentration and the embryo transfer day.

Evaluation of corpus luteum vascularization in recipient mares by using color Doppler ultrasound

Background: Embryo transfer is one of the most commonly used reproductive biotechnique. The success of embryotransfer is also affected by the synchrony of estrus and ovulation between donor and recipient animals. In horse reproduction, ultrasonography has been used, among other purposes, to diagnose early pregnancy. However, only the color Dopplerimaging mode makes it possible to evaluate the vascular architecture and the hemodynamic aspects of the vessels in severalorgans, especially the corpus luteum. The objective of this study was to evaluate, based on the color Doppler ultrasound,the corpus luteum vascularization and function from recipient mares at embryo transfer timing.Materials, Methods & Results: Mangalarga Machador mares from 5 to 10-year-old and a range of live weights of between350 to 450 kg were used for this experiment, kept in pasture-based on mombaça grass (Panicum maximum) and were givenad libitum access to water and mineral supplementation. The animals (n = 15) were gynecologically examined and uterineconsistency was evaluated by rectal palpation the same operator using an ultrasound system (SonoScape®) with a lineartransducer, and operating frequency ranging from 5 to 10 Mhz. The uterine tone was classified between grades 1 and 4 andsubjected to ovulation induction. The objective and subjective vascular perfusion of the corpus luteum was evaluated bycolor Doppler ultrasound on the day of embryo transfer and endometrium. The determination progesterone concentrationon the day of the embryo transfer was performed by direct chemiluminescence assay. The arcsine (√P/100) transformation was applied to the percentage data, and the results were expressed as mean (.) ± standard error of the mean (SEM).Further, the assumptions of normality and homoscedasticity were verified, respectively, based on the Shapiro-Wilk and...(AU)

Evaluation of corpus luteum vascularization in recipient mares by using color Doppler ultrasound

Background: Embryo transfer is one of the most commonly used reproductive biotechnique. The success of embryotransfer is also affected by the synchrony of estrus and ovulation between donor and recipient animals. In horse reproduction, ultrasonography has been used, among other purposes, to diagnose early pregnancy. However, only the color Dopplerimaging mode makes it possible to evaluate the vascular architecture and the hemodynamic aspects of the vessels in severalorgans, especially the corpus luteum. The objective of this study was to evaluate, based on the color Doppler ultrasound,the corpus luteum vascularization and function from recipient mares at embryo transfer timing.Materials, Methods & Results: Mangalarga Machador mares from 5 to 10-year-old and a range of live weights of between350 to 450 kg were used for this experiment, kept in pasture-based on mombaça grass (Panicum maximum) and were givenad libitum access to water and mineral supplementation. The animals (n = 15) were gynecologically examined and uterineconsistency was evaluated by rectal palpation the same operator using an ultrasound system (SonoScape®) with a lineartransducer, and operating frequency ranging from 5 to 10 Mhz. The uterine tone was classified between grades 1 and 4 andsubjected to ovulation induction. The objective and subjective vascular perfusion of the corpus luteum was evaluated bycolor Doppler ultrasound on the day of embryo transfer and endometrium. The determination progesterone concentrationon the day of the embryo transfer was performed by direct chemiluminescence assay. The arcsine (√P/100) transformation was applied to the percentage data, and the results were expressed as mean (.) ± standard error of the mean (SEM).Further, the assumptions of normality and homoscedasticity were verified, respectively, based on the Shapiro-Wilk and...

Functional assessment of diluent choice for semen cryopreservation from stallions with high and low freezability

Background: Fertility using horse frozen-thawed semen remains lower than in other livestock species. This fact suggeststhat horse semen hold intrinsic sensitivity to cryoinjury that must be investigated. Moreover, there is substantial evidenceof genetic factors upon horse cryopreservation outcome. Nonetheless, diluent and cryoprotectant choice for horse semencryopreservation are under intense research. Thus these factors could be explored to identify conditions that may increasesemen viability after thawing. The aim of this work was to evaluate the effect of diluents Botu-Crio®,Lactose-EDTA®, andINRA-82® on cryopreserved semen from stallions with high (HFA) and low freezability (LFA).Materials, Methods & Results: Frozen-thawed semen was evaluated for motility and membrane integrity using computerassisted semen analysis (CASA), and also inferred for sperm DNA fragmentation by sperm chromatin structure assay during the thermoresistance test (TRT). Comparisons for each parameter were done in a pair-wise fashion between HFA andLFA semen at one-hour intervals during the TRT (0 h - 4 h). Sperm motility in HFA, regardless of the diluent, was larger(P 0.05)at 0 h and 3 h. Sperm DNA fragmentation was lower (P < 0.05) in HFA semen at 0 h and 1 h.Discussion: Artificial insemination in horses using frozen-thawed semen is gaining wider acceptance under commercialsettings, although its current limited outreach due to low semen viability after thawing. Therefore, several efforts weremade toward ...

Functional assessment of diluent choice for semen cryopreservation from stallions with high and low freezability

Background: Fertility using horse frozen-thawed semen remains lower than in other livestock species. This fact suggeststhat horse semen hold intrinsic sensitivity to cryoinjury that must be investigated. Moreover, there is substantial evidenceof genetic factors upon horse cryopreservation outcome. Nonetheless, diluent and cryoprotectant choice for horse semencryopreservation are under intense research. Thus these factors could be explored to identify conditions that may increasesemen viability after thawing. The aim of this work was to evaluate the effect of diluents Botu-Crio®,Lactose-EDTA®, andINRA-82® on cryopreserved semen from stallions with high (HFA) and low freezability (LFA).Materials, Methods & Results: Frozen-thawed semen was evaluated for motility and membrane integrity using computerassisted semen analysis (CASA), and also inferred for sperm DNA fragmentation by sperm chromatin structure assay during the thermoresistance test (TRT). Comparisons for each parameter were done in a pair-wise fashion between HFA andLFA semen at one-hour intervals during the TRT (0 h - 4 h). Sperm motility in HFA, regardless of the diluent, was larger(P < 0.05) than LFA, both on 0h and 1h. In the 2h evaluation, sperm motility using Botu-Crio® and Lactose-EDTA® wasgreater (P < 0.05) for HFA. Analysis of sperm membrane integrity was similar between HFA and LFA semen (P > 0.05)at 0 h and 3 h. Sperm DNA fragmentation was lower (P < 0.05) in HFA semen at 0 h and 1 h.Discussion: Artificial insemination in horses using frozen-thawed semen is gaining wider acceptance under commercialsettings, although its current limited outreach due to low semen viability after thawing. Therefore, several efforts weremade toward ... (AU)

Freezing of Stallion Semen: In Vitro Evaluation of Motility and Acrosin Activity in Sperm Cells Cryopreserved Using Different Semen Extenders.

The work described here aimed to verify the efficiency of different extenders for cryopreservation of equine semen using sperm motility and acrosin activity as spermatic parameters. The semen was fractioned into two equal parts and resuspended in an 11% lactose solution in a 1:1 proportion, where it remained for 20 minutes at room temperature. The semen was centrifuged at 600 g for 10 minutes, and after the second centrifugation, each pellet received the freezing extender (Merck or Zorlesco) and was loaded into 4 mL straws. Each straw was placed in liquid nitrogen vapor steam for 15 minutes and further immersion in liquid nitrogen at -196°C for long-term storage. After thawing, semen samples were initially evaluated for sperm motility, both total and progressive, and acrosin activity. Moreover, semen was incubated at 37°C and further assessed at 60 and 120 minutes in a thermoresistance test (TRT) for sperm motility and acrosin activity. Immediately after thawing, both progressive and total motility, and acrosin activity were lower (p < 0.05) in thawed semen than in fresh semen. During the TRT, total sperm motility and acrosin activity after 60 minutes were lower (p < 0.05) than those obtained after thawing. Similarly, total sperm motility and acrosin activity were lower (p < 0.05) after 120 minutes than at 60 minutes of the TRT. The analysis of motility and acrosin activity allowed the conclusion that both extenders have a similar capacity to preserve the integrity of sperm cells subject to freezing and thawing.

Induction of ovulation in Mangalarga Marchador mares by hCG or GnRH

Background: Induction of ovulation is a key procedure for horse assisted reproduction technologies, such as for artificial insemination (AI) and embryo transfer. The application of hCG remains as the primary ovulation-inducing agent for horse assisted reproduction, but alternatives are in demand to avoid its adverse effects, such as loss of ovulation-inducing efficiency over multiple applications by hCC-antibody production. Despite reports on alternative ovulation-inducing agents, pair-wise comparisons of such agents under similar conditions have been limited. Under such scenario, the work was aimed to determine the efficiency of both hCG and Buserelin at inducing ovulation in Mangalarga Marchador mares raised in the Northeast of Brazil under an AI program.Materials, Methods & Results: Mares were initially selected based on their reproductive performance, the absence of clinical-reproductive alterations and adequate body condition score. Mares in diestrus were randomly distributed in three experimental conditions, received 5 mg of Dinoprost and were monitored daily for estrus detection. After estrus detection, ovaries were monitored by ultrasonography, in 12-h intervals, until the follicle reached 35 mm. At this time-point, ovulation was induced with 0.042 mg of Buserelin endovenously, with 3,000 IU hCG by an intramuscular shot, and control mares received 2 mL of saline solution, also by an intramuscular shot. Both hCG and Buserelin displayed similar efficiencies (P > 0.05) for induction of ovulation and that both agents were effective (P 0.05) from those found in the Control. All ovulations occurred within a 72-h period after treatment.[...]

Induction of ovulation in Mangalarga Marchador mares by hCG or GnRH

Background: Induction of ovulation is a key procedure for horse assisted reproduction technologies, such as for artificial insemination (AI) and embryo transfer. The application of hCG remains as the primary ovulation-inducing agent for horse assisted reproduction, but alternatives are in demand to avoid its adverse effects, such as loss of ovulation-inducing efficiency over multiple applications by hCC-antibody production. Despite reports on alternative ovulation-inducing agents, pair-wise comparisons of such agents under similar conditions have been limited. Under such scenario, the work was aimed to determine the efficiency of both hCG and Buserelin at inducing ovulation in Mangalarga Marchador mares raised in the Northeast of Brazil under an AI program.Materials, Methods & Results: Mares were initially selected based on their reproductive performance, the absence of clinical-reproductive alterations and adequate body condition score. Mares in diestrus were randomly distributed in three experimental conditions, received 5 mg of Dinoprost and were monitored daily for estrus detection. After estrus detection, ovaries were monitored by ultrasonography, in 12-h intervals, until the follicle reached 35 mm. At this time-point, ovulation was induced with 0.042 mg of Buserelin endovenously, with 3,000 IU hCG by an intramuscular shot, and control mares received 2 mL of saline solution, also by an intramuscular shot. Both hCG and Buserelin displayed similar efficiencies (P > 0.05) for induction of ovulation and that both agents were effective (P < 0.05) for such purpose, since greater percentages (P < 0.05) of induction on mares treated from those of the control. Moreover, the total number of ovulations in mares treated at the end of the experiment was not different (P > 0.05) from those found in the Control. All ovulations occurred within a 72-h period after treatment.[...](AU)