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

Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011.
A135 OPU-IVP AND ET
EFFECT OF THE METHOD FOR THE RETRIEVAL OF CUMULUS-OOCY
US-OOCYTE COMPLEXES ON THE IN VITRO PRODUCTION
EFFICIENCY OF SHEEP EMBRYOS
Maurício Barb
arbosa Salviano
1 , Cristiano Feltr
eltrin
2 , Juliana Lop
opes Almeida
2 , Kaio Cesar Simiano
Tavar
ares
2 , Felip
elipe e Jesus Mor
oraes Junior
2 , Luiz Fer
ernando Schütz 2 ,
Saul Gaudêncio Net
eto 2 , Leonar
eonardo
Tondello Mar
artins
2 , Vic
ictor Hugo Vieir
ieira Rodr
drigues
2 , Raquel Rib
ibeir
eiro Freir
eire 2 , Tatiane Mar
aria Mon
onte Nobr
obre 2 , José Baptista
Sant´Ana Neto 2 , Priscila Ferreira Alencar Silva 2 , José Luis Rigo Rodrigues 1 , Luciana Relly Bertolini 2 & Marcelo Bertolini 2
1
LABORATÓRIO DE EMBRIOLOGIA E BIOTÉCNICAS DE REPRODUÇÃO DA FACULDADE DE VETERINÁRIA DA UFRGS, PORTO ALEGRE, RS, BRAZIL. 2 LABORATÓRIO DE BIOLOGIA
MOLECULAR E DO DESENVOLVIMENTO DA UNIFOR, FORTALEZA, CE, BRAZIL.
Obtaining high quality oocytes in small ruminants for the IVP of embryos is usually a challenge. Recovery methods may
significantly affect the quality of cumulus-oocyte complexes (COCs), especially the cumulus cell vestment. The aim of this study was to compare
the efficiency of follicular aspiration and ovary slicing for the retrieval of COCs for the IVP of sheep embryos. Sheep ovaries collected at a local
slaughterhouse were transported to the laboratory in PBS (34.3±1.7 o C), being separated into two groups for the COC retrieval by follicular
aspiration or ovary slicing. For the aspiration, 2 to 8 mm follicles were aspirated using a 21G needle coupled to a 5 mL syringe, with the follicular
contents placed into 15 mL tubes. For the slicing, ovaries were placed in PBS supplemented with 1% FBS, 25 µg/mL heparin, and 50 µg/mL
gentamicin, on a Petri dish. The ovarian cortex was randomly scarified using a scalpel blade, followed by tissue washing in PBS medium. The
ovarian contents from both groups were examined under a stereomicroscope to search for COCs, which were morphologically graded as I to IV
(Leibfried & First, 1979, J Anim Sci 48:76-86). All COCs were used for the IVP of embryos using commercial media (Nutricell ® , São Paulo).
After 25 h of IVM, COCs were subjected to IVF using frozen ram semen segregated using Percoll ® gradient (a pool of three rams, with 1.5
million sperm cells/mL). After 24 h co-incubation, zygotes were in vitro-cultured for six days at 38.5 o C, 5% CO 2
and saturated humidity. Data
were analyzed by the Student´s t test or by the ÷ 2 test (P < 0.05). After four replications, a total of 328 COCs were recovered from 32 ewes
(10.2 COCs/female), with 131 (8.2 COCs/female) obtained by follicular aspiration and 197 (12.3 COCs/female) by ovary slicing. The COC
recovery rate by slicing was 50.4% higher, with a greater proportion of Grades I and II COCs, than follicular aspiration (74/197, 37.6% vs..
23/131, 17.6%, respectively). However, in spite of the similar rates of embryonic developmental between the slicing (53/197; 26.9%) and
aspiration (35/131; 26.7%) procedures, the higher number of COCs obtained by ovary slicing was reflected into a comparative increase
(51.4%) in the number of viable embryos on Day 6 by ovary slicing (3.3 embryos/female) than by follicular aspiration (2.2 embryos/female).
In addition, no differences were observed regarding stage of development and embryo quality on Day 6 between groups. In summary, the
retrieval efficiency of viable COC per female was higher after ovary slicing than follicular aspiration, which was translated into a higher ovine
IVP embryo yield on a per female basis.
Keywords: sheep, ivp of embryos, oocyte.
A136 OPU-IVP AND ET
EFFICIENCY OF OPU-IVP-ET OF FRESH AND VITRIFIED EMBRYOS IN BUFF
UFFAL
ALO
Wilson Pardini Saliba 1 , Roberti M Drumond 1 , Henrique X Bayão 1 , Múcio Túlio Alvim 1 , Pietro Sampaio Baruselli 2 , Lindsay Unno Gimenes 2 , Rômulo C Leite 3 ,
Eduardo Bastianetto 3 & Bianca Gasparrini 4
1
CENATTE EMBRIÕES LTDA, PEDRO LEOPOLDO, MG, BRAZIL. 2 FMVZ-USP, SAO PAULO, SP, BRAZIL. 3 UFMG, BELO HORIZONTE, MG, BRAZIL. 4 UNINA, NAPOLI, ITÁLIA.
OPU and IVP are promising techniques in buffalo, inversely to recovery of in vivo embryos which is still unsatisfactory. Scientific
studies show that buffaloes are able to respond to superovulatory protocols, however, embryo recovery is low (Baruselli et. al., 1999).
Moreover, success in the transfer of fresh embryos (ET) has not reached pregnancy rates higher than 15% (Sa Filho et al., 2005). The present
study aims to report OPU-IVP-ET outcomes of fresh and vitrified embryos in buffalo. For this purpose, 36 buffalo donors were submitted to
11 OPU sessions (n = 201) between July and December 2010. 998 total oocytes (5.0 ± 0.5 / donor / session) and 584 viable oocytes (2.9 ± 0.3
/ donor / session) were recovered. Viable oocytes (grades 1, 2 and 3) were subjected to IVM, IVF (D0) and IVC. On day 2 (D2) after IVF 54.5%
of cleavage rate was obtained. Embryo production was evaluated on days 5 (D5), 6 (D6) and 7 (D7) after IVF. Embryo yield was 44.9% (grade
1: 229 embryos, grade 2: 5 embryos and grade 3: 28 embryos). 115 fresh and 21 vitrified embryos were transferred into recipients previously
synchronized with ETFT protocol, 49 of the 115 ET were done with 5 days-embryos (Group D5) to synchronized 6 days after ovulationrecipients,
and 66 of the 115 ET were done with 6 days-embryos (Group D6) to 7 days after ovulation-recipients. On 30th day, 46.9% (23/49)
of pregnancy of 5 days-embryos, and 40.9% (27/66) of 6 days-embryos were confirmed. On 60th day, 44.9% (22/49) of pregnancy of 5 daysembryos,
and 39.4% (26/66) of 6 days-embryos were confirmed. Only two pregnancy losses (4.2%), one in each group, were found. No
significant differences (P > 0.05) were found between groups D5 and D6 on pregnancy rates at 30 or 60 days, and on pregnancy loss between
30 and 60 days (by chi-square test in SAS). From vitrified embryos, 33.3% (7/21) of pregnancy rate at 30 days, and 28.6% (6/21) after 60 days
of gestation (14.3% pregnancy loss) was verified. No abortion (between diagnosis of pregnancy and birth to 60 days) and stillbirth (14 births
so far) occurred until present moment. Results are unprecedented in science and opens possibilities for the commercial use of the techniques of
OPU, IVP and ET of fresh and vitrified embryos in buffaloes. [Acknowledgements: FAPEMIG (APQ - 0591-5.04/08), FINEP, FIEMG].
Keywords: ivp, et, buffalo.
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