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.
ExoELISA-ULTRA Complete Kit (CD63 detection) |
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| EXEL-ULTRA-CD63-1 | SBI | 96 reactions | 713 EUR |
ExoELISA-ULTRA Complete Kit (CD81 detection) |
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| EXEL-ULTRA-CD81-1 | SBI | 96 reactions | 713 EUR |
scanning thermometer |
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| T8710 | Consort | ea | 1282 EUR |
scanning thermometer |
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| T8720 | Consort | ea | 1282 EUR |
Multi-Colored Traceable Timer |
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| LE9002 | GenDepot | Ea | 88 EUR |
scanning thermometer ch plug |
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| T8710-CH | Consort | ea | 1282 EUR |
scanning thermometer uk plug |
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| T8710-UK | Consort | ea | 1282 EUR |
scanning thermometer ch plug |
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| T8720-CH | Consort | ea | 1282 EUR |
scanning thermometer uk plug |
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| T8720-UK | Consort | ea | 1282 EUR |
Water-Resistant, Traceable Flashing Timer |
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| LE9004 | GenDepot | Ea | 80 EUR |
thermometer probe pt100, 20m cable |
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| T4420 | Consort | ea | 154 EUR |
Traceable® Three-Line Alarm Timer |
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| LE9003 | GenDepot | Ea | 107 EUR |
thermometer probe pt100, 15cm, 20m cable |
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| T4421 | Consort | ea | 183 EUR |
thermometer probe pt100 ht, 2m cable |
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| T4422 | Consort | ea | 247 EUR |
4L Ecoscint Ultra |
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| NAT1388 | National Diagnostics | 4L | 237 EUR |
20L Ecoscint Ultra |
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| NAT1390 | National Diagnostics | 20L | 946 EUR |
CHAPS, Ultra Pure |
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| C0660-005 | GenDepot | 5g | 165 EUR |
CHAPS, Ultra Pure |
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| C0660-010 | GenDepot | 10g | 253 EUR |
CHAPSO, ultra Pure |
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| C0661-001 | GenDepot | 1g | 165 EUR |
Agarose Ultra Pure |
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| BA01101 | Bioatlas | 100g | 123 EUR |
Agarose Ultra Pure |
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| BA01102 | Bioatlas | 500g | 333 EUR |
CHAPS, Ultra Pure |
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| 22024-100G | Biotium | 100G | 584 EUR |
CHAPS, Ultra Pure |
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| 22024-10G | Biotium | 10G | 136 EUR |
Traceable® Four-Channel Alarm Timer with Clock |
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| LE9001 | GenDepot | Ea | 93 EUR |
100-Well Polycarbonate Freezer Boxes, 1" |
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100-Well Polycarbonate Freezer Boxes, 2" |
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| LC3012-012 | GenDepot | 12/CS | 253 EUR |
EpiQuik MeDIP Ultra Kit |
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| P-1052 | EpiGentek | 48 Reactions | 1005.55 EUR |
DTT, Ultra Pure, 15g |
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| R7634-15g | ACTGene | 190 EUR | |
DTT, Ultra Purer, 500g |
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| R7634-500g | ACTGene | 2846 EUR | |
DTT, Ultra Pure, 50g |
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| R7634-50g | ACTGene | 443 EUR | |
EDTA, Ultra Pure, 1kg |
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| R9410-1kg | ACTGene | 190 EUR | |
Tris - Ultra Pure Grade |
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| TB0194 | Bio Basic | 500g | 74.36 EUR |
SDS, 99% Ultra Pure |
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| S0800-100 | GenDepot | 1Kg | 313 EUR |
SDS, 99% Ultra Pure |
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| S0800-250 | GenDepot | 2.5 Kg | 536 EUR |
SDS, 99% Ultra Pure |
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| S0800-500 | GenDepot | 5 Kg | 955 EUR |
