16 t h International Congress on Animal <strong>Reproduction</strong> 196 Poster Abstracts were <strong>in</strong>sem<strong>in</strong>ated with thawed sexed semen. Motile spermatozoa were obta<strong>in</strong>ed by centrifugation on a Percoll gradient (45-90%) diluted to 1x10 6 /mL (IVF) of four different ejaculates of each bull, previously evaluated. The presumptive <strong>de</strong>nu<strong>de</strong>d zygotes, without the cumulus oophorus cells, were transferred to culture droplets of synthetic oviductal fluid medium ad<strong>de</strong>d of BSA. On Day 7 after IVF, embryos classified as Gra<strong>de</strong> 1 were counted. The rate of viable embryos was calculated by the number of Gra<strong>de</strong> 1 obta<strong>in</strong>ed embryos divi<strong>de</strong>d by viable oocytes. The total rate of viable embryos differed (p
16 t h International Congress on Animal <strong>Reproduction</strong> Poster Abstracts 197 Kg) of zoo (ZOO SAFARI-Fasano-BR). Ovaries were kept <strong>in</strong> physiological sal<strong>in</strong>e and ma<strong>in</strong>ta<strong>in</strong>ed at 35°C before oocyte recovery. Ovary were placed <strong>in</strong> phosphate buffered sal<strong>in</strong>e (PBS) supplemented with 50 micrograms/ml of gentamic<strong>in</strong>. Each ovary was sliced repeatedly with scalpel bla<strong>de</strong> to release cumulus-oocyte complex (COCs) <strong>in</strong> a 90 mm culture dish conta<strong>in</strong><strong>in</strong>g PBS at 37°C. COCs were immediately fixed <strong>in</strong> formal<strong>in</strong> (2%) and they rema<strong>in</strong> over-night at 4°C temperature. Subsequently oocyte were collect and fixed on sli<strong>de</strong> two each. Only one oocyte was collect by leopard ovaries. Oocytes was sta<strong>in</strong>ed with Hoechst 33258 and observed at fluorescence microscope (E600 Nikon, 365 nm excitation) for assess the meiotic status. Evaluation of oocytes diameter was done by stereomicroscope (Nikon ECLIPSE 50 I). Results Cat oocyte external diameter (100 micron on average) was smaller than <strong>in</strong> leopard (230 micron on average), however the nucleus-cytoplasm ratio is approximately the same 0,13 micron (14,17 micron GV diameter Vs 109 micron oocyte diameter for cat’s oocyte) vs 0,17 micron (50,13 micron GV diameter Vs 229,65 micron oocyte diameter for leopard oocytes). Conclusions Though the effective size of leopard oocytes is greater than cat’s oocytes the proportion of oocytes share are keeped. Is only the first study that shown the size of oocyte <strong>in</strong> leopard, and other research nee<strong>de</strong>d. Knowledge of morphology and size is important for subsequent <strong>in</strong> vitro maturation (IVM) an <strong>in</strong> vitro fertilization (IVF) for ma<strong>in</strong>tenance of genetic diversity <strong>in</strong> dangerous species. P514 Correlation analyses of a porc<strong>in</strong>e <strong>in</strong> vitro production system for prediction of blastocyst rates Petersen, KM*, Avery, B; Schmidt, M; Bogh, IB Veter<strong>in</strong>ary Obstetrics and R, Faculty of Life Sciences, University of Copenhagen, Denmark The aim was to f<strong>in</strong>d the best predictor for blastocyst formation <strong>in</strong> a standard porc<strong>in</strong>e IVP system. For that purpose blastocyst rates were compared with 2-pronuclear-, polyspermy- and cleavage rates, where rates were calculated over total number of IVF oocytes, and the data subjected to correlation analyses (GraphPad Prism 3), un<strong>de</strong>r the assumption that when two variables vary together, there is a correlation between them. The correlation coefficient r, which ranges from – 1 to + 1, quantifies the direction and magnitu<strong>de</strong> of correlation, and how well X and Y vary together. The best way to <strong>in</strong>terpret the value of r is to calculate r 2 , which ranges from zero to one, and is the fraction of the variance <strong>in</strong> the two variables that is shared. For example, if r 2 =0.25 then 25 % of the variance <strong>in</strong> X can be expla<strong>in</strong>ed by variation <strong>in</strong> Y. L<strong>in</strong>ear regression assumes that the data are l<strong>in</strong>ear, and f<strong>in</strong>ds the l<strong>in</strong>e that best predicts Y from X, accord<strong>in</strong>g to the equation Y = αX + B. An r 2 value of 0 means that there is no l<strong>in</strong>ear relationship between X and Y, where an r 2 value of 1 <strong>in</strong>dicates a perfect correlation. The oocytes were collected from slaughterhouse sow ovaries, IVM was performed for 44 h <strong>in</strong> TCM-199 with EGF, hCG, eCG and 10 % ECS, IVF for 24 h <strong>in</strong> a Krebs-R<strong>in</strong>ger bicarbonate based solution with 2 mM caffe<strong>in</strong>e and 0.6 % BSA, supplemented with washed semen, orig<strong>in</strong>at<strong>in</strong>g from fresh ejaculates <strong>in</strong> exten<strong>de</strong>r, and IVC for a week <strong>in</strong> PZM with 5 % ECS. IVM and IVF <strong>in</strong>cubations were done un<strong>de</strong>r 5 % CO 2 <strong>in</strong> 95 % air, and IVC <strong>in</strong> 5 % CO 2 , 5 % O 2 , and 90 % N2 at 38.5 º C. Blastocyst and cleavage rates were assessed at day 6 post <strong>in</strong>sem<strong>in</strong>ation, penetration rates after fixation 24 h post IVF <strong>in</strong> acid methanol and subsequent Orce<strong>in</strong> sta<strong>in</strong><strong>in</strong>g. In conclusion, none of the comb<strong>in</strong>ations showed perfect correlation; however the most precise predictor for blastocyst formation was the 2-pronuclear rates, which showed a significant correlation, whereas the correlations for polyspermy or cleavage were very weak (low r 2 and high P). The weak correlation for the cleavage rate is probably due to the fact that cleaved embryos are a mix of 2-PN and polyspermic oocytes, which pulls <strong>in</strong> opposite directions with regard to blastocyst formation. P515 Influence of oocyte and embryo transport <strong>in</strong>terval time on efficiency and commercial profitability of a large bov<strong>in</strong>e <strong>in</strong> vitro embryo production scale Rodrigues, JL 1 *, Queiroz, LM 2 , Feltr<strong>in</strong>, C 1 , Peixer, M 2 , Malard, P 2 , Santana, G 2 , Xavier, M 2 , Rodrigues, B 1 1Laboratory of Embryology and Biotechnics of Reprod, Faculty of Veter<strong>in</strong>ary Medic<strong>in</strong>e, Fe<strong>de</strong>ral University of Rio Gran<strong>de</strong> do Sul, Brazil; 2 Animal Biotechnology, BIO, Brazil Introduction Dur<strong>in</strong>g the last 5 years the extraord<strong>in</strong>ary <strong>in</strong>crease <strong>in</strong> the number of Nelore females submitted to ultrasound gui<strong>de</strong>d follicular aspiration (ovum pick up - OPU) <strong>in</strong> Brazil, has lead<strong>in</strong>g to the <strong>de</strong>velopment of different estrategies to overcome logistical barriers and achieve profitable pregnancy rates after transfer of IVF embryos. The objective of this experiment was to evaluate the <strong>in</strong>fluence of <strong>in</strong>terval time transport on viability of oocytes and embryos: Group 1 (G1) 1 to 2 h; Group 2 (G2) 3 to 5 h; Group 3(G3) 6 to 9 h and Group 4 (G4) 10 to 16 h. Materials and Methods Oocytes were collected by OPU (zero time), washed <strong>in</strong> modified PBS solution, and loa<strong>de</strong>d <strong>in</strong>to plastic cryovials conta<strong>in</strong><strong>in</strong>g modified TCM199 HEPES ma<strong>in</strong>ta<strong>in</strong>ed at 38°C to provi<strong>de</strong> oocyte <strong>in</strong> vitro maturation (IVM) dur<strong>in</strong>g the transport period to the laboratory. At arrival to the laboratory oocytes were transferred to modified TCM199 and cultured at 39ºC <strong>in</strong> a 100% humidified atmosphere conta<strong>in</strong><strong>in</strong>g 5% CO2. The <strong>in</strong> vitro fertilization procedure was <strong>in</strong>itiated 24 hours after OPU, and dur<strong>in</strong>g the next 24 hours the oocytes were <strong>in</strong>cubated with the spermatozoa <strong>in</strong> the same conditions as <strong>de</strong>scribed above. After that, the presumptive zygotes were transferred to modified SOFaa medium and then cultured for 7 days at the same IVM conditions un<strong>de</strong>r atmosphere conta<strong>in</strong><strong>in</strong>g 5% CO2, 5% O2 and 90 N2 %. On day 7 the embryos were evaluated, morphologically classified and loa<strong>de</strong>d <strong>in</strong> 0,25 ml straws conta<strong>in</strong><strong>in</strong>g modified SOFaa HEPES supplemented with BSA and kept at 38°C. The embryos were transferred <strong>in</strong>to previously synchronized recipients and pregnancy diagnosis was performed by ultrasound 60 days later. Statistical analysis was performed by the Chi-square test (P