Leopoldo Mayer Freitas Neto

Migration time of the genital tubercle in caprine and ovine fetuses: Comparison between breeds, sexes and species

The aim of this work was to determine the ideal moment to sex goat and sheep fetuses, to compare the average time of genital tubercle (GT) migration between sexes, breeds and species, and to evaluate the accuracy of fetal sexing between sexes. A total of 317 fetuses of 219 pregnant females were monitored at 24-hour interval, from days 30 to 60 of pregnancy in ewes, and from days 40 to 60 in goats. Examinations were performed using transrectal ultrasound equipped with a linear transducer of double frequency. Fetuses were identified as male when the GT was next to the umbilical cord and female when the GT was next to the tail. The average time of GT migration in ewes (41.3 +/- 3.1 days) was shorter (P < 0.05) than in goats (47.2 +/- 2.3 days)? In goats, the average time of GT migration of Saanen fetuses was later (P < 0.05) than in fetuses of other breeds, with no difference in the average time of GT migration between male (46.9 +/- 2.2) and female fetuses (47.4 +/- 2.4). In ewes, the average time of GT migration did not differ (P > 0.05) among breeds and sexes. In goat and sheep, no difference was noticed in the accuracy of fetal sexing between males and females (P > 0.05). The results show that fetal sexing in ewes must be done earlier than in goats, fetal sexing in Saanen goats must be performed later, and fetal sex does not influence the time of GT migration in either of the two species.

Comparison of vitrification and conventional freezing for cryopreservation of caprine embryos.

The experiment aimed to compare conventional freezing and different vitrification protocols for cryopreservation of caprine embryos at morphological, ultrastructural, and functional levels. Caprine embryos produced in vivo were allocated randomly to three groups: (1) conventional freezing with ethylene glycol (EG); (2) dimethyl sulfoxide + EG (DMSO/EG) vitrification; and (3) dimethylformamide + EG (DMF/EG) vitrification. All groups were scored for cell viability (propidium iodide staining and ultrastructural levels) and re-expansion rate after thawing or warming. Embryos subjected to DMSO/EG vitrification showed higher cell viability (73.33%), compared with DMF/EG vitrification and conventional freezing group embryos (40.00 and 66.66%, respectively). The ultrastructural study revealed that vitrified embryos had greater preservation of cellular structure than embryos from conventional freezing with EG. DMSO/EG vitrification resulted in higher rates of re-expansion in vitro (47.36%) than DMF/EG vitrification (31.58%), and conventional freezing (25.00%). In conclusion, caprine embryos produced in vivo are better cryopreserved after vitrification than conventional freezing, therefore we conclude that DMSO/EG vitrification is the most effective protocol for cryopreservation.

Conceptus loss in Santa Inês ewes carrying twin pregnancies by natural mating or embryo transfer

Commercial application of reproductive biotechnologies such as multiple ovulation and embryo transfer depends on its overall efficiency. Sheep embryo transfer is gradually gaining wider adoption, but pregnancy rates after embryo transfer remain lower than those derived from natural mating for most breeds. The work was aimed to evaluate embryonic and fetal losses in Santa Inês ewes carrying twin pregnancies by natural mating or embryo transfer. Ewes were subjected to synchronized natural mating by ram effect or used as recipients for embryo transfer. Ewes diagnosed as carrying twin pregnancies at day 25 were used in the experiment (n = 42). Conceptus viability was monitored by ultrasonography on days 30, 35, 40, 45, 50, and 55 after conception. Conceptus loss was similar (P > 0.05) within natural mating 11/42 (26.19%) and embryo transfer 14/42 (33.34%). However, overall embryonic loss (80.0%) was greater (P < 0.05) than fetal loss (20.0%), with no difference within groups The results allow the conclusion that conceptus loss after embryo transfer is similar to natural mating and occurs predominantly during the embryonic stages.

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.

Role of Male effect on Reproductive Efficiency of Nulliparous Santa Inês and Morada Nova ewes raised in Different Regions

Background: The male effect is an attractive strategy to increase herd production by concentrating mating events and deliveries and further allowing the adoption of genetic improvement programs. It holds similar efficiency to those chemically based estrous synchronization methods, but has the advantage of being a natural method. The work was aimed to evaluate the influence of male effect on estrous induction and synchronization, pregnancy and prolificacy of nulliparous Santa Ines and Morada Nova ewes raised in Semiarid and Zona da Mata regions of Pernambuco state. Materials, Methods & Results: Santa Ines (n = 80) and Morada Nova (n = 80) females, with age from 11 to 12 months, after being evaluated and selected, were identified with plastic ear tags, weighted and maintained isolated from males, during 30 days before experiment onset, without any physical, visual, olfactive and auditive contact. Estrous events were observed twice a day (6:00 and 16:00 h) by trained personnel, during a breeding season of 60 days, and estrous were considered synchronized when detected, within first five days of breeding season. Rams of Santa Ines (n = 2) and Morada Nova (n = 2) breeds were selected based upon reproductive capacity by an andrology exam, and were marked on the externum bone region with a wax and ink (4:1) mixture, and were marked in female lots in order to identify females in estrous. After ten days of breeding season onset, rams were again marked with the same wax and ink mixture, but with a different ink color. Pregnancy diagnosis was performed on day 30 by ultrasonography and confirmed on day 60 after the last mating. The statistical analysis was performed using SAS, version 8. Differences of 5% (P 0.05). The total pregnancy was 42.50% on first, 64.70% on second service and total delivery was 86.20% singletons, 12.06% twins and 3.33% triples with prolificacy of 1.15 ± 0.31. On both Semiarid and Zona da Mata regions, the majority of estrous events occurred between the11th and 15th day of the breeding season for Santa Ines ewes and between 6th and 10th day for Morada Nova ewes. Discussion: The occurrence of estrous, for both breeds, in both regions, were detected throughout the breeding seasons, despite most estrous detections were within the initial fifteen days, which normally happens with cycling pluriparous females, in disagreement with findings in the literature that young females display lower reproductive performance on the first breeding season. However, the sexual inexperience of young females is not equivalent to lack of male receptivity, since then, could not be responsible for late estrous onset in a breeding season of young females. In agreement with this statement, and based on the data described here, it has been described that young ewes display estrous within the initial 18 days of breeding season onset. The estrous dispersion in biostimulation programs is normally due to female cyclicity, a physiological condition that lowers the sensibility to estradiol negative feedback response, but are still responsible to the presence of males.