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Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011. A051 FOLLICULOGENESIS, OOGENESIS AND SUPEROVULATION EFFECT OF LOW LEVEL LASER IRRADIA ADIATION ON MITOCHONDRIAL MEMBRANE POTENTIAL AND CYCLE CLE OF BOVINE CUMULLUS CELLS Carlos Alexandre Soares 1 , Kelly Annes 1 , Thiago Revers Dreyer 1 , Herculano Silva Martinho 2 , Samir Saldanha Nicolau 2 , Fernanda Maria Sevciuc 2 , Mayra Elena Assumpção 2 & Marcella Pecora Milazzotto 1 1 UNIVERSIDADE FEDERAL DO ABC, SANTO ANDRÉ, SP, BRAZIL. 2 UNIVERSIDADE DE SÃO PAULO, SÃO PAULO, SP, BRAZIL. Bovine in vitro embryo production aims to improve livestock desirable characteristics. However, the technique efficiency must be improved. An alternative to reach this goal may be the laser therapy. It works by using low level laser irradiation (LLLI) in biological samples to generate biomodulation; however its effect on female gametes is unknown. The objective of the study was to evaluate whether treatment with LLLI is able to promote changes in bovine cumullus cells in vitro cultured. Bovine ovaries were obtained from a local slaughterhouse. Follicles (2-8mm) were aspirated and the COCs were classified according to morphology. COCs (grades I and II) were irradiated with LLL (γ633nm) by using a dose of 1J/cm 2 . After that, they were IVM (TCM-199 media, 10% bovine fetal serum and hormones) in incubator at 38.5°C, 5% CO 2 and high humidity. The control group received no irradiation. After 30 min (t30) or 24 h (t24) from the beginning of IVM, cumullus cells were mechanically removed from oocytes . After removal, cells were washed in PBS calcium and magnesium free and divided into two groups. The first group was incubated with 35uL of JC-1 (14.3ug/mL) for mitochondrial membrane potential evaluation. The second group was fixed in 70% ethanol and subsequently incubated with 50ug/mL of propidium iodide for the cell cycle evaluation. Data were collected by a flow cytometer Guava Easy Cyte Minib and graphs were evaluated by using Flojo software (v8. 7). Statistical analysis was performed using SAS system for Windows. Samples were classified as high or low mitochondrial membrane potential and G0/G1, S and G2/M of the cell cycle. There was no difference in mitochondrial membrane potential and cell cycle between between the control group and t30 (P = 0.633 and P = 0.86, respectively). However, after 24 h of IVM an increase of cells with high mitochondrial membrane potential was observed for the treated group when compared to control (P = 0.066). Regarding the cycle, there was an increase in the number of cells in both group S and G2/M. Moreover, there was a decrease in the number of cells in G0/G1 (P < 0.01). Based on these results, we concluded that the LLLI (γ633nm) with dose of 1J/cm 2 was able to induce changes in the metabolism of bovine cumullus cells cultured in vitro, which was responsible for an increased mitochondrial membrane potential and an increase of cells progressing in the cell cycle stages. Keywords: bovine, cumullus cells, low level laser. A052 FOLLICULOGENESIS, OOGENESIS AND SUPEROVULATION EFFECTS OF OVARIAN ORTOTOPIC OPIC AUT UTOTR TRANSPL ANSPLANT ANTATION TION IN REPRODUCTIVE BEHAVIOR IN GOATS (CAPR APRA A HIRCUS) Nayar ara a Almeida do Car armo mo, Samuel Amor morim Mar arcondes ondes, Fer ernanda Araujo dos San antos os, Gabr abriela Hémylin Fer erreir eira Mour oura, Talyta Lins Nunes unes, Valér aléria Ver eras de Paula, Raimundo Alv lves Bar arrêt rêto Junior unior, Michelly Fer ernandes de Mac acedo & Mar arcelo Barb arbosa Bezer erra UNIVERSIDADE FEDERAL RURAL DO SEMI-ÁRIDO, MOSSORO, RN, BRAZIL. Ovarian autotransplantation has been in use for wide goals related to reproduction in animals and humans. Once goats constitute experimental models for ovarian transplantation, this investigation aimed to evaluate the reproductive behavior of goats submitted to ortotopic ovarian autotransplantation. Thus, three crossbred goats were submitted to bilateral oophorectomy followed by autotransplantation of ovarian cortical fragments into infundibulum, ipsilateral to the donor ovaries. Fragments measuring approximately 3 x 1 x 1 mm were selected according to number and quality of preantral follicles and small antral follicles observed with the aid of an stereomicroscope (40X); after sedation, three fragments were transplanted into the infundibulum and sutured with nylon monofilament 6-0 with needle (black nylon, monofilament 6-0, Brasuture, São Sebastião da Grama, SP, Brazil). At an interval between 60 and 130 days post-transplantation, the reproductive behavior of goats was evaluated by the presence of a buck to observe the presumptive occurrence of estrus signs and possibility for mating. The results demonstrated that the first signs indicated similar behavior of proestrus that on average occurred at f 62 ± 3 days post-transplantation, accompanied by Flehmen reaction followed by courting, as well as mating attempts by the buck. This behavior was repeated at irregular intervals of 10 ± 6 days, ranging from 4 to 23 days, without mating acceptance by the goats. However, at 122 and 125 days post-transplantation, mating was observed in two goats, with one of them demonstrating persistent tail twitching and marked responsiveness to the presence of the buck prior to mating. This experiment demonstrated that ovarian tissues transplanted into the infundibulum underwent resumption of ovarian activity, but it is still reasonable to verify what variables may influences the estrus signs. More results are in progress. Keywords: autotransplantation, goat, fertility. s362
Acta Scientiae Veterinariae, 2011. 39(Suppl 1): Abstracts - 25th Annual Meeting SBTE-Brazil. August 2011. A053 FOLLICULOGENESIS, OOGENESIS AND SUPEROVULATION EFFECT OF ECG ON OVULATION AND CONCEPTION RATES IN RED SINDHI COWS ( (BOS TAUR URUS US INDICUS) TREATED TED WITH TWO SYNCHRONIZA ONIZATION OF OVULATION PROTOC OCOLS OLS Raquel Rodr drigues Costa Mello ello, Joaquim Esquer squerdo Fer erreir eira, Ana Paula Toledo Barb arbosa Silv ilva, Leandr eandro Mendes Masc ascar arenhas enhas, Ber ernar nardo Janella Fer erreir eira Silv ilva, Beatriz Oliveira Cardoso, Helcimar Barbosa Palhano & Marco Roberto Bourg Mello UFRRJ, SEROPEDICA, RJ, BRAZIL. The Red Sindhi zebu breed has excellent adaptability to adverse weather conditions and good management and ability to milk production. Although much research will be developed with these animals, studies of the reproductive traits are still scarce. Therefore, the objective of this study was to evaluate the effect of eCG on ovulation and conception rates of Red Sindhi cows during a treatment for synchronization of ovulation. Sixteen Red Sindhi cows with body condition score = 3.5 (scale of 1 to 5) were divided into two protocols for synchronization of ovulation. In protocol I, animals (n = 8) received on day 0 an intravaginal P4 device and an application of 2mg of estradiol benzoate (EB). On day 8, the device was removed and 10mg of Dinoprost (PGF2α) were applied, and on day 9, 1 mg of EB was applied, and the animals were fixed-time inseminated 24 h later; in protocol II, animals (n = 8) were submitted to the same treatment, and on day 8, 400 IU of eCG were applied. From day 0, all animals were evaluated daily by trans-rectal ultrasound device in order to monitor the follicular growth. The results were analyzed by ÷2 test, with significance level of 5%. It was observed that there was no statistical difference (P > 0.05) between protocols I and II concerning to ovulation rate, with 75% (6/8) and 100% (8/8), respectively. Regarding to conception rate, it was observed that there was statistical difference (P > 0.05) between protocols I and II (0% and 50%, respectively). The final conception rate in this study was low (25%), and this fact may be related to the high age of cows (average of 10 years old) and the long period in which they were no pregnant since they were being used as embryo donors. However, it can not conclude that treatment with eCG was efficient on the ovulation rate and conception rate in cows of Red Sindhi breed, once those are preliminary results, more research is needed on the reproductive physiology with a greater number of animals of this breed. Keywords: follicular growth, zebu, ftai. A054 FOLLICULOGENESIS, OOGENESIS AND SUPEROVULATION TIMING EFFECT OF INSEMINATION USING SEX-SORTED SPERM IN EMBRYO PRODUCTION WITH NELORE ( (BOS INDICUS) ) SUPEROVUL OVULATED DONORS Júlia Gle ley ci Soar oares 1 , Nelcio Ant onio Tonizza Car arv alho 2 , Pietr ietro Sampaio Bar aruselli 3 , Claudine laudiney Melo Mar artins 4 , Mar arcílio Nichi 5 , Manoel Francisco Sá Filho 6 , José Ricardo Garla Maio 7 , Ana Lúcia Abreu Silva 8 & Alessandra Corallo Nicacio 9 1,3,5,6 DEPARTAMENTO DE REPRODUÇÃO ANIMAL, FMVZ-VRA-USP, SAO PAULO, SP, BRAZIL. 2 UNIDADE DE PESQUISA E DESENVOLVIMENTO DE REGISTRO/PÓLO REGIONAL DO D.S.A. DO VALE DO RIBEIRA/APTA, REGISTRO, SP, BRAZIL. 4 FERTILIZA CONSULTORIA ANIMAL, POÇOS DE CALDAS, MG, BRAZIL. 7 OURO FINO SAÚDE ANIMAL, RIBEIRÃO PRETO, SP, BRAZIL. 8 MESTRADO EM CIÊNCIA ANIMAL, UNIVERSIDADE ESTADUAL DO MARANHÃO (CCA/UEMA), SÃO LUÍS, MA, BRAZIL. 9 UNIVERSIDADE FEDERAL DO PAMPA, SÃO GABRIEL, RS, BRAZIL. The effect of the moment of TAI with sex-sorted sperm in embryo production of Nellore (Bos indicus) superovulated donors was evaluated. To this aim, 16 cycling donors were used, aged between 4 and 10 years, and postpartum around 40 days. The animals were sorted evenly into four groups, according to the moment of TAI (12 and 24 vs.. 18 and 30h after administration of pLH) and type of sperm used (conventional vs.. sex-sorted). The groups were sorted as follows: TAI with conventional sperm 12 and 24h (CSG12/24) and, 18 and 30h (CSG18/30); TAI with sex-sorted sperm 12 and 24h (SSG12/24) and, 18 and 30h (SSG18/30). The design was completely randomized, in a 2x2 factorial scheme, with all the animals in their respective categories, submitted to all treatments (randomized cross-over) in a total of four replicates, with an interval of 42 days between replicates and 16 repetitions per treatment. Donors were synchronized with intravaginal progesterone device (P4) for 7 days. On the day of device insertion (D0) 2.0mg i.m. of Estradiol Benzoate were administered. From D4 to D7, all donors were treated with 8 decreasing doses of FSH (133mg i.m.; 12/12h). On D6, all animals received two PGF2α injections (0.53mg i.m.) after each FSH injection. At the last FSH dose (D7) the P4 device was removed and, donors received the pLH injection 12h later (25mg i.m.; D8). The embryonic structures were collected seven days after LHp injection (D15) and, embryos were quantified and classified according to morphology and quality (IETS, 1998). The variables were analyzed by Proc GLM procedure of SAS ® . In groups GSC12/24, GSC18/30, GSS12/24 and GSS18/30 were recovered, respectively: total of embryonic structures (2.9±1.1, 3.4±0.8, 2.6±1.2, 4.1±1.3); unfertilized oocytes (0.0±0.0, 0.1±0.1, 1.7±1.2, 0.4±0.2); transferable embryos (2.6±1.0, 2.7±0.7, 0.5±0.3, 2.2±1.0); freezable (2.4±1.0, 2.7±0.7, 0.5±0.3, 2.0±1.0) and degenerate (0.3±0.1, 0.7±0.4, 0.4±0.3, 1.5±0.8). There were no differences between groups (P > 0.05). As there was no interaction between treatments, it was possible to evaluate the main effects (type of sperm type and moment of insemination). As verified between groups, no difference was found for any of the analyzed variables between the main effects (P > 0.05). The results did not confirm the hypothesis that a delay in six hours on the FTAI with sex-sorted sperm in superovulation protocols results in increased production of viable embryos obtained from Nelore donors. Keywords: nelore, sex-sorted sperm, fixed-time superovulation. N s363
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