2011 (SBTE) 25th Annual Meeting Proceedings - International ...

Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011.
A039 MALE REPRODUCTIVE PHYSIOLOGY AND SEMEN TECHNOLOGY
LYOPHILIZED SEMINAL PLASMA IMPROVES
THE MOTILIT
TILITY OF FROZEN RAM SEMEN AND INCREASES THE CLEAVAGE IN
HETEROLOGOUS IVF
Renata Casali 1 ,Fabiano Carminatti Zago 2 , Michelle Federle 1 Jessica Nora Drum 1 , Mariana Sponchiado 1 , Norton Klein 1 , Maurício Seminotti Zanetti 1 , Saul
, Gaudêncio Neto 1 , Alceu Mezzalira 1 , Ubirajara Maciel Da Costa 1 & Arnaldo Diniz Vieira 3
1
UDESC / CAV, LAGES, SC, BRAZIL. 2 EPAGRI, LAGES, SC, BRAZIL. 3 UFPEL, PELOTAS, RS, BRAZIL.
The addition of heterologous seminal plasma (HSP) to the freezing extender of ram semen improved post-thaw viability
parameters, and might be an alternative to improve intra-cervical insemination and in vitro fertilization (IVF) with frozen ram semen.
However, data regarding such conditions are still very scarce, especially for lyophilized plasma. The aim of this study was to evaluate the
effect of addition of lyophilized bovine seminal plasma (LBSP) and equine (LESP) to the freezing extender of ram semen. The evaluations
performed were: progressive motility (PM) after thaw (AT) and PM after percoll selection (PS), acrosome integrity (AI) after PS and cleavage
rates after IVF with heterologous bovine oocytes. Seminal plasma was obtained from five bulls and five stallions collected with artificial
vagina. The ejaculates were centrifuged (5000 rpm) and lyophilized with Christ Alpha 1-4 Freeze Dryer. The ram semen was collected with
artificial vagina from four Texel rams, pooled and diluted in 1 +1 in glycerolated Tris-egg yolk without seminal plasma (Tris control, TC) or
added of 600µg/mL of LBSP or 600 µg/mL of LESP. The semen was then cooled at 0.3°C / min. until to 5ºC, stabilized for one hour, loaded
in 0.25 mL straws in isothermal conditions and frozen at -80°C in LN2 vapor before store in cryogenic container at -196°C. The sperm
viability was determined based on the PM-AT and PM-PS. The sperm were used for heterologous IVF (Garcia-Alvarez, 2009, Theriogenology,
71: 643-650), and to evaluate the acrosome reaction by FITC method (Sukardi Ani Sci Rep 1997, 46: 89-96). Data were analyzed by Student
T test with significance level of 5%. There was no difference between the average rates of post thaw progressive motility PM-AT of PSLB
(27.5%) and PSLE (37.5%) groups, but both were higher than TC group (10%), improving the viability of thawed semen by the addition of
heterologous seminal plasma. The rates of PM-PS were similar in all groups (TC 40.0%, 56.7% LBSP, LESP 70.0%). There was no difference
in the IAPP evaluation of different groups (CT 58.4%, 67.3% LBSP, LESP 63.3%). There was a significant increase in cleavage rate of LBSP
(45.7%) and LESP (43.7%) groups when compared to TC group (32.6%). The increased cleavage rate observed in heterologous IVF
demonstrates that the frozen semen added of LEAP or LBSP has a greater oocyte penetration ability, which suggests that they may be more
efficient for sheep artificial insemination.
Keywords: cryopreservation, lyophilization, heterologous seminal plasma.
A040 MALE REPRODUCTIVE PHYSIOLOGY AND SEMEN TECHNOLOGY
USE OF THE POWDERED COC
OCONUT
ONUT WATER AS AN ALTERNA
TERNATIVE TIVE EXTENDER FOR THE CRYOPRESER
OPRESERVATION OF COLL
OLLARED
PECCARIES (
(TAYASSU
ASSU TAJA
AJACU
CU) ) SEMEN
Mariana Araújo Silva, , Lívia Batista Campos, José Artur Brilhante Bezerra, Gabriela Liberalino Lima, Gislayne Christianne Xavier Peixoto, Andréia Maria Da
Silva & Alexandre Rodrigues Silva
UFERSA, MOSSORÓ, RN, BRAZIL.
Collared peccaries (Tayassu tajacu) are amongst the most hunted species due to the appreciation for its meat and international
interest for his leather. This fact has contributed to a drastic decrease in its population in their natural habitats. In this sense, the development of
protocols for the storage of semen would allow the formation of germplasm banks for its use in captive breeding programs. Once the literature
is scarce, and only the Tris (Castelo et al., Cryobiology, 2010) extender has been used for this purpose, the present study aims to evaluate the
efficiency of a powdered coconut water-based extender (ACP-116c ® ) as an alternative for the cryopreservation of collared peccary semen.
Twelve adult males bred in captivity at the Centre of Multiplication of Wild Animals of UFERSA were used. They were mechanically restrained
with the aid of a hand-net, and further anesthetized with propofol (5 mg/kg, IV). The electroejaculation was conducted, and the semen samples
were evaluated for microscopic characteristics. Each sample was divided into two portions, the first was diluted in Tris-fructose and the second
in ACP-116c ® , both plus 10% egg yolk. Samples were equilibrated for 240 min at 5°C, and further added of 3% glycerol, reaching a 100 x 106
sperm/mL concentration. Samples were packed in 0.25 mL plastic straws and stored in liquid nitrogen. After one week, semen was thawed in
a water-bath at 37ºC/30s and reevaluated. Data for sperm motility were Arcsin transformed and submitted to analysis of variance followed by
Student’s t-test (P < 0.05). Sperm vigor was evaluated by the Mann-Whitney non-parametric test (P < 0.05). Fresh semen presented a
concentration of 765 ± 313.8 x 106 sperm/mL, in a volume of 2.8 ± 0.7 mL, with 86.7 ± 2.6% motile sperm with vigor 4.4 ± 0.2. After thawing,
a reduction was verified for all the parameters assessed in the use of both diluents (P < 0.05), which differed among themselves (P > 0.05).
Values observed for sperm motility was 30.4 ± 5.7% and 40.8 ± 6.9%, and for vigor was 2.4 ± 0.2 and 2.9 ± 0.2 for Tris and ACP-116c ® ,
respectively (P > 0.05). The results evidence an equivalence of diluents on the preservation of sperm quality. The use of ACP-116c ® is
recommended as an alternative extender for the collared peccary semen cryopreservation.
Keywords: powdered coconut water, cryopreservation, Tayassu tajacu.
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