Effect of cryopreservation on sperm DNA fragmentation and apoptosis rates in the testicular tissue of domestic cats.

The objectives of the present study were to evaluate the damage caused by cryopreservation on sperm DNA and estimate the percentage of cell apoptosis in tissue after thawing. Testicles of cats were sectioned into of 0.3 cm3 and 0.5 cm3 fragments and evaluated for DNA damage using acridine orange and semi-quantitatively through histo-morphological and immunohistochemical methods (caspase-3). Other fragments were placed in cryotubes with diluent containing either 3% glycerol or 3% propanediol, and were cryopreserved. Evaluation using acridine orange indicated there was a difference with use of propanediol and glycerol on DNA damage in 0.5 cm3fragments, with the latter being more effective than the former for cryopreservation. Results from histomorphological evaluations indicated there was a greater cell integrity among germ cells that were not cryopreserved, based on criteria assessed (detachment of cells from basal membrane, retraction of seminiferous tubule epithelium, visibility of the spermatogonia nucleoli and nuclear spermatogonia condensation), for both sizes of fragments. The values for these variables decreased after cryopreservation, with there being no differences as a result of size of fragment stored or between cryoprotectants used (P > 0.05). The staining for caspase-3 differed for the cytoplasm, nuclei and germ cells. Assessment of these staining patterns indicated the fresh fragments had an amount of cell damage and there was a similar amount of damage detected in cryopreserved fragments. This finding indicated that there was considerable efficacy in preserving the tissue fragments with use of the freezing protocols that were evaluated in this study.

An update on the cecectomy technique in roosters anesthetized with isoflurane used in subsequent amino acid digestibility experiments.

This work aims to study an alternative technique of cecectomy in roosters using inhalation anesthesia for subsequent use in digestibility experiments. A total of 30 adult chickens of Leghorn breed were used with an average age of 27 wk. The birds were preoxygenated, and the anesthetic induction was performed using isoflurane diluted in oxygen. After proper muscle relaxation, endotracheal intubation was performed using a Murphy catheter and kept in anesthesia under mechanical ventilation with a constant monitoring of electrocardiography variables, heart rate, oxyhemoglobin saturation, and body temperature during the surgical procedure. An incision of approximately 3 cm was made between keel and cloaca in order to expose and extirpate the cecum followed by a simple ligature. The opening of the peritoneal cavity was closed in 2 ways: Sultan suture technique was used for closing the abdominal wall and modified Cushing intradermic continuous points for closing the skin. The time for anesthesia induction and preoperational period noted to be 10 ± 2 min. Surgical procedures completed in 7 ± 1.5 min. No intraoperatively and postoperatively harm observed in animals. Water was immediately provided after the birds returned to cages and feed offered after 24 h of surgery. In remaining birds, no abnormalities were observed during and after the experimental period (3 mo). The present study describes a promising update on cecectomy technique regarding anesthesia induction and surgical procedures in roosters using potential drugs and safer surgical materials without any trans- and postoperative complications.