Reproduction in Domestic Animals - Facultad de Ciencias Veterinarias

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16 t h International Congress on Animal Reproduction 188 Poster Abstracts with r 2 = 0.97. The values of L P were determined to be 0.79 ± 0.26, 0.82 ± 0.22, and 0.64 ± 0.16 μm⋅min -1 ⋅atm -1 and the P CPA values were determined to be 2.9 ± 1.3, 2.7 ± 1.3, and 0.27 ± 0.18 x10 -3 cm⋅min -1 for DMSO, EG and GLY respectively. There were no significant differences (p>0.05) between values for L P and P CPA in presence of the DMSO and EG. However, these values were significantly different from the values in presence of GLY. Conclusions Similar to human oocytes, rabbit oocytes behave as ideal osmometers in the range osmolalities tested with values for V bp nearly identical to human oocytes. However, rabbit oocyte isotonic volume is smaller than human oocytes. Previously published mean values for L p in human oocytes falls within the 95% confidence limits for rabbit oocytes; the same is true for the permeability to DMSO. Rabbit oocytes are more permeable to EG in comparison to human oocytes, however. Higher P CPA values for rabbit oocytes result in less volume excursions than those for human oocyte during CPA addition and dilution. Supported by Wellcome Trust (Grant No.070246), EU FP6 (MEXT-CT-2003-509582, MRTN-CT-2006-035468) and Chinese- Hungarian Bilateral projects (TET CHN-28/04, CHN-41/05). P487 Different response of Bos indicus Vs Bos taurus oocyte on maturation, cleavage and embryo development under in vitro system Escalona, F*; Mercado, J; Rodríguez, A; Rodríguez-Sallaberry, C; Kowalski, AA Laboratorio de Embriología y Endocrinología Molecular, Decanato de Agronomía, Universidad Centroccidental Lisandro Alvarado (UCLA), Lara, Venezuela The in vitro of production embryos represents an alternative to the cattle industry for generating large numbers of F1 embryos. The objective of this study was determine the differences of in vitro maturation, fertilization and culture (IVM, IVF, IVC) of two oocytes groups; Brahman (Br) and Holstein (Ho) oocytes. Ovaries from slaughterhouse were transported in saline 0.9% at 30 ±1.1°C; complex oocytes-cumulus (COCs) were collected from Br (52) and Ho (51) ovaries and used for IVM, IVF and IVC. The number of oocytes from each ovary were: Br (13.65±1.3) Ho (7.30±3) respectively. COCs were cultured on TCM-199 supplemented with BSA, piruvate, L- glutamine, FSH, LH y EGF during 22 hrs, and incubated at 38°C on 5% CO 2 in a humidified environment. The COCs were transferred to IVF-TALP supplemented with heparin, penicillamine, hypotaurin and epynephrine. The fertilization was made with sperm from Ho bulls. The semen was separated by percoll gradient and washed in spermtalp. In both groups the semen concentration was 1 x 10 6 sperm/mL and incubated for 18 hrs. The zygotes were incubated for 7 days in KSOM supplemented with BSA, L-glutamine, EAA and NEAA. The cleavage rate was Br (92.36±0.8%) and Ho (69.4±18.7%) (P
16 t h International Congress on Animal Reproduction Poster Abstracts 189 P490 Use of immunomodulation in susceptible mares to clear an experimentally induced endometritis effect on proinflammatory cytokines: IL-1β, IL-6 and IL-8 mRNA expression Fumuso, E* 1 ; Giguère, S 2 ; Rogan, D 4 , Rivulgo, V 1 ; Wade, J 3 ; Rodriguez, E 1 ; and Sánchez Bruni S 1 1Faculty of Veterinary Medicine, UNCPBA, Tandil, Argentina 7000, Argentina; 2Coll.Vet. Med. Gainesville, FL, USA; 3 Ireland; 4 Bioniche Life Sciences Inc, Belleville, Ontario, Canada The effect of an immunomodulator, Mycobacterial Cell Wall Extract (MCWE), on IL-1β; IL-6 and IL-8 mRNA expression was studied in mares susceptible to endometritis after experimental uterine infection with Streptococcus zooepidemicus. Thirty endometritis susceptible mares, based on the presence of uterine fluid during both diestrus and estrus, were inoculated with 5 × 10 6 CFU of S. zooepidemicus on day 1 of estrus. Twenty-four hours later, the progression of infection was evaluated by ultrasonography, bacteriology, exfoliative cytology, and uterine biopsy. Forty eight hours after inoculation and confirmation of uterine infection, mares were randomly assigned to one of four unbalanced experimental groups in order to receive: Group A: 1500 μg total dose of MCWE (Settle) by intrauterine route (IU) (n = 10), Group B: the same treatment as Group A, but using the intravenous (IV) route (n = 10), Group C: Distilled water as placebo (PL) IU (n = 5) and the Group D: was treated as Group C using the IV route (n = 5). Endometrial biopsies were taken when infection was confirmed (day 0), at ovulation (day 3) and 7 days post-ovulation (day 10) for measurement IL-1β; IL-6 and IL-8 mRNA expression. Total RNA was isolated, treated with DNAse-I and cDNA was synthesized. Relative quantitation of mRNA expression (RmRNA) was determined by real-time PCR. The effect of treatment (MCWE vs PL), administration route (IU vs IV), and day of sampling (0, 3, 10) on RmRNA mixed model analysis of a split-unit experiment with repeated observations. There was no effect of route of administration on RmRNA. As a result Groups A & B (treated) and groups C & D (PL), were combined in the final analysis. There were no significant differences in RmRNA between PL and treated groups on day 0. On days 3 and 10, MCWE treated mares had significantly lower IL-1β, IL-6 and IL-8 mRNA expression than mares in the PL group. We previously demonstrated the effect of immunomodulation after AI in susceptible mares to post breeding endometritis (Fumuso et al. 2003, 2007); this effect was confirmed in the present study through experimentally induced endometritis. Results indicate that MCWE exerts an anti-inflammatory effect on cytokine RmRNA that may contribute to resolution of endometritis caused by S. zooepidemicus in mares. P491 The addition of malondialdehyde during in vitro maturation of bovine oocytes improves subsequent cleavage rate after in vitro fertilization Garcia-Ispierto, I 1 *, Leroy, JLMR 2 ; Lopez-Béjar, M 1 ; López-Gatius, F 3 ; De Clercq, JPB 2 ; Andries, S 2 ; Goovaerts, IGF 2 ; Bols, PEJ 2 1Department of Animal Health and Anatomy, Autonomous University of Barcelona, Spain; 2 Laboratory of Veterinary Physiology, Department of Veterinary Sciences, University of Antwerp, Belgium; 3 Department of Animal Production, University of Lleida, Spain Introduction Oxidative stress has been related to heat shock on embryos and has been considered to play a critical role in the success of in vitro fertilization protocols. Malondialdehyde (MDA) is the major endogenous product of lipid peroxidation due to oxidative stress. It has been demonstrated that MDA can have toxic effects on bacterial and mammalian cells. Oxidative stress is high during the postpartum period and this can be exacerbated by heat stress during the hot seasons. Objective The aim of the current study was to apply MDA combined with high temperature conditions during in vitro maturation of oocytes to evaluate possible effects on oocyte developmental competence. Methods Bovine ovaries were obtained from the slaughterhouse. A total of 1209 immature Grade I cumulus–oocyte complexes (COCs) were aspirated from follicles 2–6mm of diameter. COCs were cultured in groups of 50 for 24 h in 500µl serum-free maturation medium (20 ng/ml mEGF) with or without MDA (8µM) under normal (38.5ºC) or high temperature (41ºC) conditions in a humidified 5% CO 2 incubator. Four treatment groups were used during maturation: control (C), MDA (M), high temperature (T) and MDA plus high temperature (TM). After IVM, all oocytes were routinely fertilized by co-incubation per 100 with spermatozoa (frozen bull semen selected by percoll gradient) at a final concentration of 10 6 sperm cells/ml for 20 h at 38.5ºC in fertilization medium, in a humidified 5% CO 2 incubator. Presumptive zygotes were cultured per 25 in 500µl droplets of modified SOF medium with 5% FCS, under mineral oil for 8 days. Results Binary logistic regression procedures were performed using cleavage and blastocysts rate (blastocysts per oocytes matured) as dependent variable, and treatment and replicate as independent variables (SPSS 16.0). Cleavage and blastocyst rate were 70.5+7.5 and 29.4+3.1 in C, 80.6+8.5 and 35.5+11.3 in M, 40.8+12.9 and 4.9+3.4 in T, and 39.3+3.4 and 7.6+7.8 in TM groups, respectively. Based on the odds ratio the cleavage rate decreases in the high temperature group (by a factor of 0.27, P
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