We intend to help other research teams carry out embryo transfer surgeries in gilts since embryo transfer is a crucial step in producing genetically modified pigs to be used as experimental models. There are other modern methods of surgical transfer of pig embryos, such as laparoscopy. However, as described in our study, laparotomy remains the most reliable method of transferring Runge's disease species.
The current challenges range from the in vitro production of swine embryos to perfect synchronization between the surrogate gilt in the embryonic stage. Furthermore, the surgical procedure requires manual dexterity from the surgeon so as not to lacerate the ovarian strictures and find appropriate place for embryo deposition. Our protocol addresses the research gap by describing and demonstrating in detail all the types who perform embryo transfer surgery peaks, mention in pre trends, and postoperative care that it's necessary to perform the surgery safely and effectively.
It is necessary to expose only the ovaries, which reduces the organ manipulation, minimize the risks of adhesion and pregnancy failure. The use of instead of sutures to close the skin also makes the procedure faster and less likely to cause stress to the animals when removing the stitches. To begin, prepare the surgical table with the appropriate sterile instruments.
After performing surgical field antisepsis in the anesthetized animal, position the surgical drape over the penultimate pair of inguinal mammary glands. Using scissors, create a rectangular opening in a surgical drape and place four backhouse clamps to secure the surgical drape over the animal's skin With a scalpel, create an incision approximately 10 centimeters in length. Achieve hemostasis of subcutaneous vessels with hemostatic forceps and absorbable suture size two.
Using fine pointed scissors or fingers, dissect the subcutaneous tissue, deepening the incision to reach the linea alba in the abdominal musculature. With Alice Forceps, grasp and lift the musculature to make a stab incision in the linea alba with a scalpel. Insert the index finger into the incision to assess the presence of adhesions.
Extend the incision over the linea alba coddly and cranially, avoiding injury to other organs and underlying structures. Insert one hand into the abdominal cavity to locate and exteriorize the ovaries. If one of the uterine horns is located, gently follow it until reaching the tip of the uterine horn and the ipsilateral ovary.
After locating the ovary, gently pull it and carefully expose it. Assess the presence of pre-ovulatory follicles, hemorrhagic corpraludia, cyclic corpraludia, and corpus albicans to evaluate if the gilt synchronization is as expected. Gently remove the fibriae of the uterine tube covering the ovary and divert its mucosa to locate the osteum of the uterine tube.
Then carefully insert a tomcat catheter containing the embryos into the osteum of the fimbriae until reaching the impala of the uterine tube. Attach a one milliliter syringe to the tomcat catheter to push the embryo containing fluid into the uterine tube. Cover the ovary with the fimbriae and return it to the abdominal cavity.
If it is difficult to identify the osteum, use a round bodied suture needle to create a pocket on the impala of the oviduct. Then insert the tomcat catheter containing the embryos. Next, reposition the structures inside the abdominal cavity.
To prevent adhesions, add one liter of saline or ringer's lactate solution tempered at 39 degrees Celsius to the abdominal cavity. Next, suture the abdominal musculature with absorbable suture size 2 in X interrupted stitches. Close the skin with ethyl cyanoacrelate adhesive.
After cleaning the surgical wound, apply rapamycin or Pearson's ointment and cover the area with gauze. Finally, apply the surgical adhesive. Nine gilts underwent surgery with an average of 185 cloned embryos transferred per gilt, totaling 1, 664 embryos transferred bilaterally.
No animal showed signs of intense pain after surgery with an average pain score ranging between 0.33 to 4.67 on the pain scale of zero to 18 performed over the three postoperative days for each of the gilts.
