Laparoscopic embryo transfer in pigs - comparison of different variants and efficiencies of the method.

The aim of the study was to develop a method of laparoscopic embryo transfer in pigs and to compare different variants of this method. Two catheter diameters (1.6 mm and 1.0 mm), the method and site of embryo deposition (oviduct or uterus), the embryo development stage (2 - 4 cell or blastocyst), the method for oviduct or uterus stabilization, the potential for cryopreserved embryo transfer, the developmental potential of the embryos after transfer to the oviduct, patomorphology of the oviduct after transfer and possible clinical complications were taken into consideration. Two studies compared two variants of transfer to the uterus, and five variants of transfer to the fallopian tube. The transfer of embryos by the infundibulum may be of limited use due to handling problems and very low efficiency (pregnancy was not achieved). Very low efficiency was shown after transfer of vitrified embryos. Transfer to the fallopian tube by puncture of the fallopian tube, regardless of the developmental stage of the embryo, is the recommended method of embryo transfer. The histopathological examination of the fallopian tube revealed possible changes within the puncture site. The numerous clinical complications observed did not affect the effectiveness of the method.

The repeatable method of laparoscopic ovum pick-up (OPU) in sheep: clinical aspects and efficiency of the method.

The aim of the study was to develop new laparoscopic technique for repeated recovery of sheep oocytes. Oocytes were aspirated with specifically designed catheter. It allowed to recover oocytes without ovary damage and to preserve very good quality of recovered oocytes. Fifteen ewes were oocytes donors. Oocytes were collected: one time (group I, n=15), two times (group II, n=15), three times (group III, n=10), four times (group IV, n=5). The endoscope was inserted into the abdominal cavity. Two trockars for putting the manipulators were inserted 15 cm cranial from the udder. Oocytes were collected by aspiration of the follicular fluid from the ovarian follicles. The observed clinical complications were: ovary bleeding and cicatrix at place of needle insertion, the fragmentary adhesion of infundibulum and ovary, adhesions of omentum and peri- toneum near the place where the grasping forceps were inserted and adhesion of ovary and uterus. Ovarian follicles (n=204) were aspirated, 130 (63.8%) oocytes were obtained. Out of 130 obtained oocytes, 112 were qualified for in vitro maturation. The remaining 18 oocytes (13.8%) were rejected due to cytoplasmic changes. The proposed technique allows for the collecting oocytes of good quality that can be used for IMV/IVF techniques and cloning.

Piglets born after intrauterine laparoscopic embryo transfer.

The aim of the study was the preliminary development of laparoscopic transfer of embryos to the uterus in the pig, which can become the alternative for more invasive surgical methods. We proposed the original method of embryo transfer. Donors (n = 40) and recipients (n = 15) of embryos were sows of age of 6-8 months. The estrus cycle of both recipients and donors was routinely synchronized. The experimental animals were divided into two groups. In the first group (10 donors and 3 recipients) embryos were transplanted according to the method described earlier and in the second group (30 donors and 12 recipients) embryos were transplanted according to our own proposed method. As the control group, we used 16 sows after insemination (AI). In animals from both experimental groups pregnancy was diagnosed between 28-31 day after transplantation and in the control group between 28-31 day after insemination. All animals were observed during pregnancy and weaning period in pig farm. Embryos at the development stage of 2-4 cell were obtained surgically and cultured in vitro for 4 days. Obtained blastocysts were transferred to donors. The original set of catheters for blastocysts transfer to pig uterus was constructed. Three trocars were placed in abdominal cavity for inserting endoscope and 2 grasps for uterus stabilization. After uterus stabilization, the slide was inserted into abdomen which was used for putting the needle to puncture uterus. Through this needle catheter with embryos was inserted into the uterus cavity. Embryos were placed by injection into lumen of the one uterine horn. From 12 recipients pregnancy was diagnosed in 6 recipients. From 6 litters, 57 piglets were born. We weaned 41 piglets (71.9%). In our study we obtained 50% efficacy, with the mean number of 9.5 alive piglets in litter and 6.8 weaned piglets. The efficacy of developed method of laparoscopic transfer of porcine embryos allows it to be used in routine practice.