Single fixed-time laparoscopic intrauterine insemination as a tool to obtain low-diversity porcine embryos
2013
Brussow, K-P., Leibniz Inst. for Farm Animal Biology, Dummerstorf (Germany). Inst. of Reproductive Biology | Vernunft, A., Leibniz Inst. for Farm Animal Biology, Dummerstorf (Germany). Inst. of Reproductive Biology | Kempisty, B., University of Medical Sciences, Poznan (Poland). Dept. of Histology and Embryology | Ratky, J., Research Institute for Animal Breeding and Nutrition, Herceghalom (Hungary)
The aim of the present study was to reduce embryo diversity and to achieve a uniform outcome of porcine embryo stages using single laparoscopic fixed-time insemination (LIUI). Altogether, 48 puberal German Landrace gilts were included in the study. Estrus of gilts was synchronized by 15-day long altrenogest (Regumate) feeding and follicle development was stimulated with 850 IU eCG 24 h after the final altrenogest application. Ovulation was induced with 500 IU hCG 80 h after eCG. LIUI was performed 31 h after hCG treatment. Gilts under general anaesthesia were fixed in a dorsal position, a pneumoperitoneum was produced and three trocar cannulas were inserted into the abdomen for optics and instruments. Each uterine horn was carefully punctured 10-15 cm caudal from the utero-tubal junction with a 2.5 mm trocar. A 2.2 mm catheter was inserted about 3 cm into the uterine lumen and 20 ml of extended fresh boar semen (32.2 x 106 sperm cells/ml) were injected. Embryos were surgically flushed from the genital tract two (Day 2) and three (Day 3) days after insemination. Altogether, 778 oocytes/embryos were recovered (recovery rate 68 +/- 17%); 45 of 48 gilts (93.8%) revealed fertilisation and 76.1% of the recovered embryos (n = 592) were at the 2- and 4-cell stage. On Day 2 (n = 22 gilts), a higher percentage of gilts (72.7%, P less than 0.05) displayed only 2-cell embryos compared with gilts which had 2- and 4-cell (22.7%), or only 4-cell embryos (4.6%). On Day 3 (n = 23 gilts), the proportion of gilts with 2-cell, 2- and 4-cell, and only 4-cell embryos shifted to 4.3%, 0% and 95.7%, resp. (P less than 0.05). The results of the present study demonstrate high rates of fertilisation and homogenously developed embryos after single fixed-time laparoscopic intrauterine insemination in gilts. Additionally, these results were achieved by inseminating a 60% lower number of sperm cells per insemination dose compared to usual doses used for transcervical insemination. In conclusion, LIUI can be recommended for the in vivo production of embryos in a homogeneous developmental stage, and also as an alternative method for low-dose insemination.
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