Successful short term sperm cryopreservation in brown-marbled grouper (Epinephelus fuscoguttatus) with the utility of ultra-freezer (-80 ℃)
Production of cryopreserved semen in fish generally requires liquid nitrogen (LN), which is not always easily available in remote areas. To reduce reliance on LN, the aim of the present study was to evaluate if electric freezer could be a feasible LN-free alternative to cryopreserve brown-marbled grouper sperm. After loading, semen straws were put directly in freezers (-30 or -80 ℃) for frozen and then transferred to LN for storage.
Compared with the conventional LN vapor freezing (straws were put horizontally 3 cm above the surface of LN), there was a significant reduction in all tested post-thaw sperm quality parameters in samples frozen at -30 ℃ for 10 min, including kinetic parameters (total motility: 85.0% vs 48.6%), viability (84.7% vs 51.7%), high mitochondrial membrane potential (86.4% vs 63.7%), ATP content (106.9 nM/109 cells vs 72.9 nM/109 cells) and hatching rate (86.3% vs 45.7%), accompanied with an increasing lipid peroxidation level (MDA content: 11.9 nM/109 cells vs 4.9 nM/109 cells).
In contrast, frozen with -80 ℃ ultra-freezer (10 min or 12 h) produced similar sperm quality parameters to those using LN, except that temporary storage (12 h) at -80 ℃ yielded lower average path velocity. In conclusion, this study confirmed that -80 ℃ ultra-freezer is an effective alternative to LN for sperm freezing in brown-marbled grouper.
Physiological, biochemical, and hormonal changes in rats exposed to bottled water left in a hot car and the freezer
The aim of this study was to investigate the effects of bottled water left in the car and those left in the refrigerator on the physiological and biochemical parameters of male rats. Eighteen male albino rats were randomly divided into control, warmed bottled water (WBW), and cooled bottled water (CBW) groups. The concentrations of bisphenol A (BPA) and antimony (Sb) in the water samples were measured.
Results showed that the levels of BPA and Sb were higher in bottled water left in the car and those kept at room temperature (control), relative to the bottled water samples kept in the fridge. Bodyweight, some metabolic indicators, cTnT and CRP levels, hematological parameters, testosterone level, and sperm quality were significantly affected by storage conditions.
These results indicate that storage conditions can potentially affect the quality of bottled water, which in turn can affect the biological parameters of living organisms. Moreover, since the concentrations of BPA and Sb were least in bottled water kept in the refrigerator, we recommend that manufacturers and households should endeavor to keep bottled water under cold storage.
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