Influence of Vitrification Device, Warming Protocol, and Subsequent In Vitro Culture on Structural Integrity of Testicular Fragments from Adult Domestic Cats.

The objective of the study was to evaluate the integrity of cat testicular tissues after vitrification with different devices followed by different warming conditions. The influence of vitro culture for 24 hours after warming also was examined. Testicular tissues from adult domestic cats were dissected in small fragments that were vitrified using Cryotop® or threaded on fine needles, warmed (directly at 37°C or with a preliminary 10 seconds exposure to 50°C), and/or cultured in vitro for an additional 24 hours. For each treatment group, tissues were assessed based on histology, apoptosis, and sperm DNA integrity. Results showed that fragments of testicular tissues were efficiently cryopreserved (maintaining the quality of all cell types) with vitrification with Cryotop followed by direct warming at 37°C, and additional culture of 24 hours at 38.5°C. These encouraging results are paving the road to optimize preservation protocols and use them for systematic banking of tissues from genetically valuable felids.

Ontogenetic development of the oral apparatus and oropharyngeal cavity in bullfrog tadpoles (Lithobates catesbeianus, Shaw 1802).

OBJECTIVE: The present study aimed to describe the morphology of oral apparatus and oral cavity of bullfrog tadpoles during their development and metamorphosis. DESIGN: The oral apparatus and oropharyngeal cavity of tadpoles from hatching up to metamorphosis stage was dissected for further analysis. These structures were fixed in Karnovsky solution, afterwards in osmium tetroxide and metalized in palladium gold and electron-micrographed using the scanning electron microscope. RESULTS: The development of oral apparatus started with the formation and keratinization of the jaw sheaths and labial teeth followed by the formation of marginal and sub-marginal papillae. Degeneration of oral apparatus and formation of mouth was observed during metamorphosis. From stage-42 (metamorphic climax) to stage-43, the jaw sheath and labial tooth rows were disappeared progressively while the size and number of labial papillae were decreased. At stage-44, mouth formation started with the development of anterior and posterior labium though the labial papillae were still present. At stage-45 and 46, mouth was already formed, being very similar to the adult and characterized by the progressive increase in size. CONCLUSION: The sequence of events that happen during the development of oral apparatus of Lithobates catesbeianus Shaw, 1802 tadpoles follows the same pattern as occur in other anuran species but metamorphic atrophy of the oral apparatus follows the sequence of morphogenesis.

Cat epididymal semen cryopreserved with and without vitamin E: effect on sperm parameters and lipid peroxidation.

The aims of this study were to investigate: 1) if the addition of α-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of α-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) - epididymal sperm were frozen with a commercial Botucrio® extender; group 0.3, group 0.6 and group 0.9 - the extender was supplemented with 0.3, 0.6 and 0.9 mM of α-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three α-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the α-tocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of α-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of α-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages.