The laparoscopic method of recovering oocytes in goats and sheep is one of the minimally invasive methods used in the biotechnology of animal reproduction. It allows for good quality oocytes that are suitable for in vitro maturation and fertilization to be recovered. The limitation of using the laparoscopic ovum pick-up (L-OPU) method in goat and sheep is its changing effectiveness and the lack of repeatability of results, as well as the varying effectiveness of different variants of the method. Therefore, it is necessary to develop effective non-invasive techniques allowing for multiple good quality oocyte recovery that would be suitable for in vitro maturation and fertilization. In this study, four different L-OPU variants were described in goats and sheep. Various techniques of recovering oocytes were discussed, including the techniques of conducting the operation, various tools for recovering oocytes, and different plans of hormonal stimulation. Recovery rates were 35% (Variant I), 57% (Variant II), 72% (Variant III), and 67% (Variant IV). After evaluation, 94% (both Variant I and II), 93% (Variant III), and 84% (Variant IV) of the oocytes were qualified for in vitro maturation. The results of the study show that the proposed technique of laparoscopic recovery of oocytes allows a sufficient number of ovarian cells suitable for in vitro culture to be obtained and as a consequence it makes them useful in in vitro maturation/in vitro fertilization (IVM/IVF) programs or cloning. The method allows for a fast and effective conduct of the operation in a living donor with minimal invasiveness while preserving the excellent condition of animals.
Freezing/thawing procedures induce enhanced reactive oxygen species (ROS) formation in mammalian sperm and these ROS may be a cause for the decrease in sperm function following cryopreservation. In the present study, we used a chemiluminescence method to detect ROS-induced damage in goat spermatozoa. Iron-induced luminescence of fresh and frozen/thawed sperm cells was assessed using a luminometer. It was shown that the freezing/thawing procedure had a significant effect on some luminescence parameters. Semen freezing significantly increased the values of integral, peak max, T.half (rise) and T.max (peak) parameters. A significant correlation was observed between the percentage of motile spermatozoa and integral, peak max and T.half (rise) parameters. In conclusion, the results of the present study indicate that measurement of induced luminescence can be an alternative, sensitive and relatively simple method for assessing the effect of cryopreservation on oxidative damage to spermatozoa.
Emergency endoscopy is usually complicated by unfavorable examination conditions. The irrigating capability of the instrument is inadequate. The use of an easy-to-operate volume- and pressure-driven irrigation pump has proved very efficient and safe in the hands of an experienced endoscopist, and helps establish the correct diagnosis or provide proper treatment.
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 peritoneum 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.
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