RESUMO
The axolotl (Ambystoma mexicanum) draws great attention around the world for its importance as a biomedical research model, but housing and maintaining live animals is increasingly expensive and risky as new transgenic lines are developed. The goal of this work was to develop an initial practical pathway for sperm cryopreservation to support germplasm repository development. The present study assembled a pathway through the investigation of axolotl sperm collection by stripping, refrigerated storage in various osmotic pressures, cryopreservation in various cryoprotectants, and in vitro fertilization using thawed sperm. By the stripping of males, 25-800 µL of sperm fluid was collected at concentrations of 1.6 × 106 to 8.9 × 107 sperm/mL. Sperm remained motile for 5 d in Hanks' Balanced Salt Solution (HBSS) at osmolalities of 100-600 mOsm/kg. Sperm cryopreserved in 0.25 mL French straws at 20 °C/min in a final concentration of 5% DMFA plus 200 mM trehalose and thawed at 25 °C for 15 s resulted in 52 ± 12% total post-thaw motility. In six in vitro fertilization trials, 20% of eggs tested with thawed sperm continued to develop to stage 7-8 after 24 h, and a third of those embryos (58) hatched. This work is the first report of successful production of axolotl offspring with cryopreserved sperm, providing a general framework for pathway development to establish Ambystoma germplasm repositories for future research and applications.
RESUMO
Noise represents a threat to human and wildlife health, triggering physiological and behavioral challenges to individuals living close to sources of extreme noise. Here, we considered airport environments as sources of potentially stressful stimuli for birds and tested if those living near airports are under higher physiological stress than birds living in quiet sites. We used measurements of CORT in feathers (CORTf) as a proxy of chronic stress. We evaluated 14 passerine and 1 non-passerine species, living near three Brazilian airports. We found that, across species, individuals with a better body condition had lower CORTf concentration. At the species level, we found that CORTf concentration was not consistently affected by airport noise. Comparing individuals living in quiet sites with those living near airports, we found that 2 species had higher and 2 had lower CORTf concentrations near airports, while 11 species presented no significant differences between sites. At the population level, model selection indicated that the direction and strength of these differences are weakly related to species' song frequency (peak frequency), as lower-frequency singers tended to present higher CORTf levels at airport-affected sites. In summary, we were unable to find a consistent response among species, probably due to species-specific differences in their response to anthropogenic disturbances. Instead, we found that species might be affected differently according to their singing spectral frequency and that individuals in good body condition show lower CORTf, suggesting that this measure is consistent with lower physiological stress.