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

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Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011. A119 OPU-IVP AND ET ULTR TRASOUND-GUIDED FOLLICLE ASPIRATION DO NOT REDUCE BLOOD FLOW IN THE FOLLICULAR WALL - PRELIMINARY RESULTS Alberto Mansur Ghetti 1 , Felipe Zandonadi 2 , Luiz Gustavo Bruno Siqueira 3 , Luiz Sérgio Almeida Camargo 4 , Eduardo Kenji N. Arashiro 5 , Carlos Alberto Soares Paim 6 & João Henrique Moreira Viana 7 1,2 UFF, NITERÓI, RJ, BRAZIL. 3,4,7 EMBRAPA CNPGL, JUIZ DE FORA, MG, BRAZIL. 5 UFMG, BELO HORIZONTE, MG, BRAZIL. 6 CENTRO DE ENSINO SUPERIOR DE JUIZ DE FORA, JUIZ DE FORA, MG, BRAZIL. Removal of the follicular content by transvaginal ultrasound-guided aspiration (OPU) may not induce immediate atresia and loss of function, resulting in the occurrence of residual or persistent follicles (Viana et al. 2001; Dorea et al., 2011). The presence of steroidogenic activity post-aspiration in these structures may compromise the efficiency of the synchronization of follicular growth by removing the dominant follicle. The mechanisms associated with maintaining or not of the activity of the remaining follicular wall cells, however, are not clear. The aim of this study was to evaluate the changes in blood flow caused by follicular aspiration in functional dominant follicles. Holstein-Zebu cows (n = 10), lactating, cyclic and with body condition score of 2.4 ± 0.36 were used. The follicular growth was synchronized (D0) by 1 mg of oestradiol benzoate (Sincrodiol, OuroFino Agronegócios, Cravinhos, SP), and insertion of a progesterone releasing device (Sincrogest, OuroFino). In D0 the cows received 0.5 mg of clorprostenol sodium (Sincrocio, OuroFino) to prevent any interference of any corpus luteum in the evaluation of vascular dynamics. The dominant follicle of the subsequent wave (mean diameter 12.9±0.8 mm) was aspirated using conventional OPU procedures. The blood flow (BF) was monitored by color Doppler immediately before and every 12 h after aspiration for three days, using an ultrasound device equipped with a 7.5 MHz linear transducer (MyLabVet 30, Esaote, Genoa, Italy). The BF of each follicle was measured by calculating the ratio of the maximum area of the Doppler signal and the area of the follicular wall in the center section of the follicle. All aspirated follicles showed persistence, characterized by initial formation of a clot in the cavity 12 h after follicular aspiration, followed by retraction and formation of fluid-filled cavity with anechoic appearance, similar to the antrum of non-aspirated follicles. In one animal (10%) a reduction (87.0%) in BF was observed subsequent to aspiration, in the other animals the BF showed a quadratic effect according time postaspiration (y= -1.16x2 +11.33x +16.26; R² = 0.76), with an increase up to 48h and a reduction thereafter (P < 0.01). The increase in BF after aspiration may be due to the inflammatory reaction associated with the procedure, and could indirectly contribute to the maintenance of the steroidogenic activity in the remaining cells of the follicular wall. The preliminary results suggest that the follicle aspiration affects the pattern of vascular wall, leading to a transient increase in blood flow. [Acknowledgment: Nutricell Nutrientes Celulares, CAPES, Fapemig and Embrapa project 01.07.01.002]. Keywords: opu, blood flow, doppler. A120 OPU-IVP AND ET EVAL ALUATION OF OOCYTES RECOVERED FROM COWS UNDERGOING IMMUNOSUPPRESSIVE PROTOC OCOL OL WITH DEXAMETHASONE André Penido Oliveira, Paula Maria Pires Do Nascimento, , Juliana Marques Bicalho, Guilherme Gomes De Carvalho, Rejane Silva Diniz, Jenner Karlisson Pimenta Dos Reis, Rômulo C Leite & Magali D´Angelo UFMG, BELO HORIZONTE, MG, BRAZIL. Glucocorticoids have immunosuppressive and antiinflammatory activity, affect the release of GnRH (Nangalama and Moberg, 1991, Journal of Endocrinol. 137, 87-94) and can affect programs of fixed-time AI, in vivo and in vitro embryo production. The objective of the study was to evaluate the production of COCs recovered by OPU after dexamethasone administration in cows. We used nine (n = 9) females, with average weight of 454 ± 55.7 kg and 3.5 ± 0.5 score of body condition, aged between four and six years old. The animals were divided evenly into two groups: treatment group (Gt, n = 6) and control group (Gc, n = 3). In the D0 Gt received 0.1 mg / kg of dexamethasone (iv), repeated every 24 h for 5 consecutive days. The Gc group received saline solution (0.1 mg / kg IV) like Gt. Both groups underwent OPU every four days (D0, D4, D8, D12, D16 and D20), in a total of six aspirations in the experimental period (20 days). COCs recovered were classified as viable or unviable, according to Viana et al. (2004, Anim. Reprod. Sci. 84: 1-12). The data were analysed by t test or correlation analysis. Difference was observed between the number of viable COCs and unviable during the experimental period. No differences were observed in the total number of COCs between days of treatment (P < 0,05). Low correlation (r = 0,07) between the number of viable COCs and treatment day, so the dexamethasone protocol did not affect the quality and quantity of recovered COCs. Keywords: oocyte, dexametason, bovine. s396
Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011. A121 OPU-IVP AND ET EVAL ALUATION OF A MEDIUM BASED ON COC OCONUT ONUT WATER (COC OCOS OS NUCIFERA) TO IN VITRO EMBRYO PRODUCTION OF BOVINE Marcela Da Silva Cordeiro 1 , Priscilla Do Carmo De Azevedo Ramos 2 , Nathália Nogueira Da Costa 2 , Thiago Velasco Guimarães Silva 2 , Priscila Di Paula Bessa Santana 2 , Bruno Baraúna Da Silva 2 , Davi César Nascimento Dos Santos 2 , Andre Luiz Alves De Sá 2 , Stefanne Dhullia Braga Conceicão 2 , Carlos Leonardo De Aragão Araújo 2 , Simone Do Socorro Damasceno Santos 2 , Moysés Dos Santos Miranda 2 & Otávio Mitio Ohashi 2 1 INSTITUTO FEDERAL DE EDUCAÇÃO, CIÊNCIA E TECNOLOGIA DO PARÁ, ABAETETUBA, PA, BRAZIL. 2 UNIVERSIDADE FEDERAL DO PARÁ, BELEM, PA, BRAZIL. Studies on the chemical composition of coconut water in natura reveal that some of its features are quite different from those presented by the TCM 199 (Ramos, et al., 2010, Acta Scientiae Veterinariae, 38: 764) which limits its use as a culture medium in IVP. Therefore the aim of this study was to evaluate a supplemented medium based on coconut water (S-BCWM) to be used in the process of IVM, IVF and IVC. The S-BCWM consisted of 20 mL of in natura coconut water (5 months of age) + 80 mL of ultrapure water + osmolytes (0.46g NaCl and 0.018g MgSO4, values determined in a previous analysis of chemical composition of coconut water in natura). Amino acids, myo-inositol, trisodium citrate and sodium bicarbonate were added according to basic formulation of the SOF (Tervit et al., 1972 Journal Reproduction Fertility, 30: 493-497). For IVM period the TCM 199 and S-BCWM media were supplemented with 0.5µg/mL FSH, 50µg/ml LH, 50µg/mL of gentamicin, 22µg/mL of pyruvate and 10% FCS. In the IVF the SOF medium (without glucose) and S-BCWM were add with 10µg/mL heparin, 2µM penicillamine, 1µM hypotaurine, 0.25 µM epinephrine, 50µg/mL of gentamicin, 22µg/mL of pyruvate and 6 mg/mL BSA. In the process of the IVC the SOF (1 mM glucose) and S-BCWM were supplemented with 6 mg/mL BSA, 10 µg/mL insulin, 50µg/mL of gentamicin and 10% FCS. COCs from slaughterhouse were selected and randomly distributed among the following groups: G1 (IVM in TCM199, IVF in IVC and IVC in SOF), G2 (IVM in S-BCWM, IVF in SOF and IVC in SOF), G3 (IVM in TCM 199, IVF in S-BCWM and IVC in SOF) and G4 (IVM in TCM199, IVF in SOF and IVC in S-BCWM). Fertilization was performed 18 h after the onset of IVM with semen from a single bull (2x10 6 sperm/mL). After 30h of IVF, presumptive zygotes were transferred to culture media drops for co-culture with a cumulus cells monolayer. IVP process was performed in incubator with 5% CO 2 , at 38.5°C with high humidity. Fertilization, cleavage and blastocyst rates were analyzed by ANOVA (significance level of 5%). It was observed that the maturation of oocytes resulted in S-BCWM Cleavage (82.50 ± 5.2%) and blastocysts (36.07 ± 8.8%) similar to those obtained by maturation in TCM 199 (82.61 ± 6.7% to 41.47 ± 7.9%, P > 0.05). However, the use of S-BCWM during the IVF period blocked the fertilization process, once 100% of the oocytes were arrested in MII stage after 18h IVF. When S-BCWM was used for embryo cultivation there was no embryonic development. Therefore, the S-BCWM can be an option only as a maturation medium in the process of IVP of bovine. Acknowledgements: FAPESPA and UNOPAR. Keywords: coconut water, cocos nucifera, ivp. A122 OPU-IVP AND ET EVAL ALUATION OF A NEW METHOD FOR IDENTIFYING ENDOMETRITIS IN ANESTROUS MARES THROUGH ULTR TRASSONOGR ASSONOGRAPHY APHY Gabriel Maksoud Greco 1 , Leticia Mendes Pupio Maia 1 , Maria Manoela Castro Chaves 2 & Marco Antonio Alvarenga 42 N 1 MÉDICO VETERINÁRIO AUTÔNOMO, CONSELHEIRO LAFAIETE, MG, BRAZIL. 2 UNIVERSIDADE ESTADUAL PAULISTA, BOTUCATU, SP, BRAZIL. At the beginning and end of the breeding season, when the percentage of recipient mares showing normal estrous cycles is low compared to donors, progesterone-treated acyclic recipients have been routinely used in embryo transfer programs. However, anestrous recipients are often selected for treatment through the evaluation of their body score condition and reproductive history, since their poor uterine tone and endometrial atrophy usually impair it´s evaluation through ultrassonography and rectal palpation. The objective of the present study was to test the hypothesis that a single estrogen injection could aid in the ultrassographical identification of endometritis in anestrous mares. Twentyfour anestrous mares that did not show air, endometrial edema or uterine fluid through ultrassography were submitted for endometrial cytology with a sterile uterine brush. A minimum of ten fields were evaluated per Giemsa-stained slide. Mares were classified as positive for cytology (moderate endometritis) when more than two neutrophils were microscopically identified per field under oil immersion (×100). Animals were subsequently injected with 5.0 mg of estradiol benzoate (Estrogin ® , Farmavet, São Paulo, Brazil) I.M., being evaluated through ultrassonography past 48 h to detect uterine fluid accumulation. The presence and depth of intra-uterine fluid was recorded. Results were statistically analyzed through the Chi-square test, significance levels set as P < 0.05. Of the twenty-four anestrous mares, eleven were positive and thirteen negative for cytology. All of them presented some grade of endometrial edema 48 h post-injection. Intra-uterine fluid was detected in all of the positive cytology mares (100%; 11/11), which was fairly echogenic in three (27.3%; 3/11) and had a depth of a least 20 mm in eight (72.7%; 8/11). On the other hand, five of the thirteen negative cytology mares (38.5%; 5/13) showed fluid accumulation and only a single mare (20.0%; 1/5) had more than 20 mm of uterine fluid. Positive cytology mares had higher chances of accumulating intra-uterine fluid, which can exceed 20 mm in depth (P < 0.05). In conclusion, mares showing uterine fluid 48 h post-treatment with 5.0 mg of estradiol benzoate should not be used as embryo recipients, for they have higher chances of suffering from endometritis. Moreover, authors suggest that this method should be routinely implemented during reproductive evaluations of anestrous recipient mares, especially prior to purchase. Keywords: mare, anestrous, endometritis. s397
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