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Repeated laparoscopic follicular aspiration in lambs
152
Ultrasonography REPEATED LAPAROSCOPIC FOLLICULAR ASPIRATION IN LAMBS Anel, L. , Sevillano, C. , Alvarez, M., Alegre, B., Anel, E. , Dominguez, J.C., Carbajo, M.T. and De La Fuente, ~1. Animal Reproduction. Fat. Vet. Le6n University. 24071, Leon, Spain. 1CIT-INIA. Reprod. Anim. P.O. Box 8111.28080 Madrid. Spain. Reduction of the generation interval is an important objetive in MOET breeding schemes. Laparoscopic folliculoaspiration (LFA) and in vitro fertilization (IVF) techniques applied to non-puber animals shortening drastically this intervals. The production of embryos following IVF of oocytes from young donors has been reported in several studies. The practical use of this technology will depend on the number of embryos produced per donor which is related in part on the number of oocytes obtained per collection. The continous aspiration of a determinate donor within limited time periods would be very advantageous from the genetic point of view. Two experimental groups with 5 lambs per group (8-9 weeks old at the start), were used in an experimental LFA program. Six LFA replicates were realized during eleven weeks in both group. Group A was treated with 1200 iu eCG (FolligonQ Intervet) 48h before LFA and 2ml monoclonal antibody anti-eCG (Neutra-PMSG@, Intervet) immediately after LFA. Group B was treated with a total dose of 4ml of FSH (Ovagen@, ICP) in a single injection diluted in polyvinylpyrrolidone (PVP-40, Sigma) administered 96h before LFA. Under general anesthesia the ovaries were grasped aided by laparoscopic observation and the follicles (2-8 mm) were punctured with a 18 G needle connected to a vacuum line (100 mm Hg). The aspiration medium was TCM-199 supplemented with heparin and antibiotic. Only COCs I, II and III were considerated as valids for this study. The results are presented in the Table 1. Table 1. Production of oocytes through continous LFA. Group An x LFA Fol.Punct.
eCG FSH
5x6 5x6
410 477
COCs Valids Total valids COC COCs recovered per Animal % per LFA n % per LFA n 36.6 210 51.2 7.0f2. 9 184 87.6. 6.1X2.7 39.6 215 45.1 7.2k1.7 198 92.1 6.6~t1.7
No significant differences were found between groups. Despite the treatment used, these results show, with an acceptable recovery rate, a high number of valids COCs recovered per donor during a limited time period. Several studies have been published with a single LFA and differents stimulation protocols (intravaginal progestagens with FSH and eCG combinations); Coonrod et al. (Theriogenology 41 : 182, 1994) report 8.8 oocytes per donor. Also in vivo maturation treatments with GnRH and FSH/LH (Earl et al., Theriogenology 43 : 203, 1995) report 10.5 and 5 oocytes recovered per lamb. In our study similar recovery rates were obtained by continous LFA (every 14d) with a simple and economic hormonal stimulation, enabling harvesting a large number of oocytes from an indivudual genetic superior donor. This study was supported in part by : Diputacion de Valladolid and Junta de Castilla y Leon.
Ultrasonography REPEATED LAPAROSCOPIC FOLLICULAR ASPIRATION IN LAMBS Anel, L. , Sevillano, C. , Alvarez, M., Alegre, B., Anel, E. , Dominguez, J.C., Carbajo, M.T. and De La Fuente, ~1. Animal Reproduction. Fat. Vet. Le6n University. 24071, Leon, Spain. 1CIT-INIA. Reprod. Anim. P.O. Box 8111.28080 Madrid. Spain. Reduction of the generation interval is an important objetive in MOET breeding schemes. Laparoscopic folliculoaspiration (LFA) and in vitro fertilization (IVF) techniques applied to non-puber animals shortening drastically this intervals. The production of embryos following IVF of oocytes from young donors has been reported in several studies. The practical use of this technology will depend on the number of embryos produced per donor which is related in part on the number of oocytes obtained per collection. The continous aspiration of a determinate donor within limited time periods would be very advantageous from the genetic point of view. Two experimental groups with 5 lambs per group (8-9 weeks old at the start), were used in an experimental LFA program. Six LFA replicates were realized during eleven weeks in both group. Group A was treated with 1200 iu eCG (FolligonQ Intervet) 48h before LFA and 2ml monoclonal antibody anti-eCG (Neutra-PMSG@, Intervet) immediately after LFA. Group B was treated with a total dose of 4ml of FSH (Ovagen@, ICP) in a single injection diluted in polyvinylpyrrolidone (PVP-40, Sigma) administered 96h before LFA. Under general anesthesia the ovaries were grasped aided by laparoscopic observation and the follicles (2-8 mm) were punctured with a 18 G needle connected to a vacuum line (100 mm Hg). The aspiration medium was TCM-199 supplemented with heparin and antibiotic. Only COCs I, II and III were considerated as valids for this study. The results are presented in the Table 1. Table 1. Production of oocytes through continous LFA. Group An x LFA Fol.Punct.
eCG FSH
5x6 5x6
410 477
COCs Valids Total valids COC COCs recovered per Animal % per LFA n % per LFA n 36.6 210 51.2 7.0f2. 9 184 87.6. 6.1X2.7 39.6 215 45.1 7.2k1.7 198 92.1 6.6~t1.7
No significant differences were found between groups. Despite the treatment used, these results show, with an acceptable recovery rate, a high number of valids COCs recovered per donor during a limited time period. Several studies have been published with a single LFA and differents stimulation protocols (intravaginal progestagens with FSH and eCG combinations); Coonrod et al. (Theriogenology 41 : 182, 1994) report 8.8 oocytes per donor. Also in vivo maturation treatments with GnRH and FSH/LH (Earl et al., Theriogenology 43 : 203, 1995) report 10.5 and 5 oocytes recovered per lamb. In our study similar recovery rates were obtained by continous LFA (every 14d) with a simple and economic hormonal stimulation, enabling harvesting a large number of oocytes from an indivudual genetic superior donor. This study was supported in part by : Diputacion de Valladolid and Junta de Castilla y Leon.