39 Nanowater enhances cryoprotective effects of glycerol during ram semen freezing
Bibliographic record
Abstract
It was suggested that unique physicochemical properties of nanowater (NW, water declustered in the cold plasma generator), such as a low dielectric constant and high diffusivity, might ameliorate ram semen freezing by improving bioavailability of extender constituents. This study was undertaken to evaluate the effects of NW added to glycerol-containing semen extenders on the characteristics of frozen-thawed ram semen. A total of 18 ejaculates collected from 6 Olkuska breed rams during the breeding season were divided into six equal portions. The ejaculates were frozen in the modified fructose-skimmed milk-egg yolk Kareta extender containing 3 or 7% glycerol and diluted in deionized water (control extenders; Aqua Purificata, Prolab; C3% and C7%, respectively) or NW (declustered for 15 min (NW15’) or 30 min (NW30’); Nanotechnology Systems) to a final concentration of 800 × 106 spermatozoa/mL. They were placed in 0.25-mL plastic straws and frozen in liquid nitrogen. The two declusterization times were chosen on the basis of previous laboratory tests and fertility trials yielding the best results in terms of semen quality post-thawing and pregnancy rates after AI. All semen samples were evaluated for sperm concentration, progressive motility (Sperm Class Analyzer), and morphological defect rates (Nikon Eclipse 80i microscope, Nikon Corp.). In addition, the ex situ survival time at 37°C and extender content of alanine transferase, alkaline phosphatase, and aspartate aminotransferase were measured, along with the proportions of apoptotic, necrotic, and live spermatozoa determined post-thawing with flow cytometry (BD Accuri™ C6 Plus, Becton Dickinson) at 0 and 1 h post-thawing. Data were analysed by one-way analysis of variance and Holm-Sidak method using SigmaPlot statistical software (Systat Software Inc.). The proportion of spermatozoa with mid-piece defects was significantly (P < 0.05) reduced in NW15’-3% compared with C3% (1.3 ± 0.3% vs. 4.0 ± 06%; mean ± s.e.m.). The proportion of necrotic spermatozoa 1 h after thawing was greater (P < 0.05) in C7% compared with NW30’-7% (20.7 ± 0.4% vs. 17.6 ± 1.0%), whereas the proportions of live cells detected immediately (0 h) and 1 h after thawing were greater (P < 0.05) in NW30’-7% than in C30’ (0 h: 54.0 ± 0.6% vs. 50.4 ± 1.2%; 1 h: 54.4 ± 1.2% vs. 59.2 ± 0.4%, respectively). The mean survival time of spermatozoa was greater (P < 0.05) in extenders dissolved in NW30’compared with their respective controls (NW30’-3% vs. C3%: 215 ± 4 min vs. 189 ± 6 min; and NW30’-7% vs. C7%: 242 ± 8 min vs. 217 ± 4 min, respectively). Alkaline phosphatase concentrations in extenders prepared with NW30’ were lower (P < 0.05) compared with the control groups (NW30’-3% vs. C3%: 3105 ± 231 IU vs. 4490 ± 458 IU; and NW30’-7% vs. C7%: 2516 ± 128 IU vs. 3956 ± 263 IU, respectively). Our results indicate that NW significantly improves the effects of glycerol on ram sperm viability post-thawing, with the reduction in sperm necrosis/overall enhancement of sperm survivability being greater with 7% compared with 3% of glycerol in semen extender.
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How this classification was reachedexpand
Full frame distilled prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.
Codex and Gemma teacher scores by category
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.000 | 0.000 |
| Meta-epidemiology (narrow) | 0.000 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.000 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.000 | 0.000 |
| Insufficient payload (model declined to judge) | 0.000 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one teacher head, not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".