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

Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011.
A127 OPU-IVP AND ET
DIFFERENT VOL
OLUMES AND CENTRIFUGATIONS
TIONS TIME OF PERCOLL GRADIENT DO NOT AFFECT THE SPERM QUALIT
ALITY AND
EMBRYO DEVELOPMENT OF IN VITRO PRODUCED BOVINE EMBRYOS
Liziane Lemos
Vianna
1 , Jor
orgea Pradieé
2 , Elisa Car
aroline Da Silv
ilva San
antos
3 , Ale
lexander Oliv
liveir
eira Gonçalv
onçalves
4 , Ledi Anghinoni
5 , Mar
aria Gabr
abriela
Tavar
ares
Rheingantz 6 , Luiz Francisco Machado Pfeifer 7 , Margot Alves Nunes Dode 8 , Marcio Nunes Corrêa 9 & Ligia Margareth Cantarelli Pegoraro 10
1,2,3,4,6,9
UNIVERSIDADE FEDERAL DE PELOTAS, PELOTAS, RS, BRAZIL. 5,10 EMBRAPA CLIMA TEMPERADO, PELOTAS, RS, BRAZIL. 7 EMBRAPA RONDÔNIA, PORTO VELHO, PORTO
VENLHO, MS, BRAZIL. 8 EMBRAPA RECURSOS GENÉTICOS E BIOTECNOLOGIA, BRASÍLIA, DF, BRAZIL.
The objective of this study was to compare the embryo development and sperm quality and sex ratio of in vitro produced bovine
embryos. Cumulus-oocyte complexes (COCs) were aspirated, from ovaries collected from slaughterhouse, selected and matured for 22 to 24h.
For sperm selection, the Percoll (Sigma, Aldrich, St. Louis, USA) was preparated in the convencional way - P Group (90 e 45% with total
volume of 4mL) and minigradient way PM Group - (90 e 45% with total volume of 800 µL). For IVF, the semen was thawed and added to the
gradients. The P Group was centrifuged for 20 min at 700 xg and the PM Group for 5 min at 700 xg. In both the second centrifugation was
performed for 5 min at 700 xg. The CCOs were randomly divided between treatments (P Group n = 830; PM Group n = 897) inseminated and
maintained for 18h in IVF medium. Briefly, they were transferred to culture medium (SOFaa) and incubated for 7 days, at 39 o C and 5% CO2.
Cleavage and embryonic global rates (delevoped embryos/inseminated oocytes) were observed at D2 and D7 respectively (Day 0 = FIV). For
this study, 11 routines of IVP were performed. Embryos of each groups (P Group n = 115; PM Group n = 102) were harvested at D8 of culture
and prepared for sexing determination by PCR. To evaluate the integrity of the plasmatic membrane we used 6-carboxiphluorescein diacetate
(FDA) with propidium iodide (PI) and to acrosome integrity fluorescein isocianate (FITC) conjugated with peanut agglutinin (PNA)
(Molecular Probe ® , Eugene, Oregon, USA). Two evaluations were performed: before (Control Group) and after treatments (3 replicates). A
chi-square analysis was performed to compare cleavage, embryonic global rates between groups and sex ratio to expected of 50%. The semen
characteristics were analyzed by one-way ANOVA using the General Linear Models (GLM) procedure. Tukey test was used to detect
differences between means. The P Group had higher cleavage rate than PM Group (70.4% P versus 65.8% PM, P < 0.05). However, no
difference was observed between groups on the embryonic development (19.2% P versus 16.3% MP). Semen quality evaluations (membrane
and acrosome integrity) did not differ between groups. The sex ratio observed was similar between treatments. The results showed no
influence of volume and time of centrifugation in the use of Percoll Gradient, on the embryo production and sperm quality.
Keywords: bovine; sex ratio; sperm selection.
A128 OPU-IVP AND ET
CUMULATIVE EFFECT OF BOVINE RECOMBINANT SOMATOTR
TROPIN (RBST) ) ON THE IN VITRO PRODUCTION OF EMBRYOS
FROM LACT
CTATING TING GUZOL
OLANDO FEMALES
Felip
elipe e Jesus Mor
oraes Junior
1,18 , Luiz Fer
ernando Schütz 1 , Cristiano Feltr
eltrin
1 , Leonar
eonardo
Tondello Mar
artins
1 , Saul Gaudêncio Net
eto 1 , Maurício Barb
arbosa Salviano
1 , Kaio
Cesar Simiano
Tavar
ares
1 , Juliana Lop
opes Almeida
1 , Vic
ictor Hugo Vieir
ieira Rodr
drigues
1 , Jamir Machado Junior
1 , Cesár Car
arneir
neiro Linhares Fer
ernades
2 , Barbar
arbara a Mar
ara
Bandeira Santos 2 , Carlos Enrique Mendez Calderón 3 , Luis Henrique Aguiar 1 , Eudes Vieira Castro 3 , Francisco Jose Magalhães Barbosa 4 , Luciana Relly
Ber
ertolini
1 , Jose Adalmir
Tor
orres Sousa
5 & Mar
arcelo Ber
ertolini
1
1
LABORATÓRIO DE BIOLOGIA MOLECULAR E DO DESENVOLVIMENTO, UNIVERSIDADE DE FORTALEZA (UNIFOR), FORTALEZA, CE, BRAZIL. 2 UNIVERSIDADE ESTADUAL DO CEARÁ -
UECE, FORTALEZA, CE, BRAZIL. 3 UNIVERSIDADE COOPERATIVA DA COLOMBIA – UCC, BUCARAMANGA, COLÔMBIA; 4 GRUPO EDSON QUEIROZ, FORTALEZA, CE, BRAZIL. 5 LAB. DE
REPRODUÇÃO ANIMAL, UNIVERSIDADE FEDERAL DO PIAUÍ – UFPI, TERESINA, PI, BRAZIL.
The aim of this study was to evaluate the cumulative effect of different doses of recombinant bovine somatotropin (rbST; Boostin ® ,
Schering-Plough) on the number and quality of cumulus-oocyte complexes (COCs) obtained in vivo from lactating Guzolando cows, along with
its effect on the in vitro production (IVP) of embryos. For that purpose, 14 half-blood Guzolando cycling multiparous females were selected at
67.3 ± 16.5 days of lactation, body condition score of 3.3 ± 0.2, and 6.6 ± 0.4 years of age. Females were randomly allocated to three
experimental groups (Control, 250 mg and 500 mg rbST) and subjected to six sessions of in vivo follicular aspiration (ovum pick up, OPU) at
a 14 day interval. Six days prior to each OPU session, a 1.5 mL dose of saline solution (Control, n = 4), 250 mg rbST (n = 5) or 500 mg rbST
(n=5) was injected in the base of the tail in each animal. Recovered COCs were morphologically Graded as I to IV (Leibfried & First 1979, J
Anim Sci 48:76-86), and subjected to IVP. Rates of in vitro embryo development to the compact morula and blastocyst stages were evaluated on
Day 6.5 of culture. The mean numbers of recovered COCs, viable COCs (Grades I, II, and III), and embryos produced by OPU-IPV per
session/female were compared by the Student’s t test, whereas the proportion of recovered COCs, based on the morphological grade, and rate
of embryo development on Day 6.5, were compared by the ÷ 2 test, for P < 0.05. When compared with the control group, the administration of
250 mg and 500 mg rbST increased the mean number of recovered COCs (8.6 ± 1.6 vs.. 13.3 ± 1.5 and 11.3 ± 1.6) and viable COCs (5.8 ± 1.1
vs.. 7.6 ± 0.8 and 6.5 ± 0.8), for a lower proportion of viable COCs (66.7% vs.. 56.8% and 57.8%), respectively. In addition, the 250 mg and
500 mg rbST groups yielded a greater mean number of Grade III (5.3 ± 0.7 and 4.8 ± 0.6 vs.. 4.0 ± 0.7) and Grade IV COCs (6.0 ± 1.0 and 4.8
± 0.9 vs.. 2.9 ± 0.6) when compared with the control group, respectively. Nevertheless, despite the increase in COCs numbers and decrease in
COCs quality seen in the 250 mg and 500 mg treatment groups relative to the controls, no differences were observed in the rates of embryo
development (80/227, 35.2%; 77 / 370, 36.0%; and 57/138, 41.3%) or the mean number of viable embryos per session/donor (2.7 ± 0.4; 2.6 ±
0.4; and 2.4 ± 0.4) between groups, respectively. In summary, our preliminary results indicated that the administration of 250 mg and 500 mg
rbST in Guzolando cows six days prior to in vivo oocyte retrieval was followed by an increase in the number of COCs, with a slight reduction
in quality, which were not translated into differences in the in vitro embryo production efficiency in any group.
Keywords: bovine, ccos, rbst.
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