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頁籤選單縮合
題名 | Cryopreservation of Spermatophores in the White Shrimp (Litopenaeus vannamei)=白蝦精莢之超低溫保存探討 |
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作者 | 趙乃賢; 陳雨農; 謝文陽; 林金榮; | 書刊名 | 水產研究 |
卷期 | 17:2 2009.12[民98.12] |
頁次 | 頁67-76 |
分類號 | 439.66 |
關鍵詞 | 超低溫冷凍保存; 抗凍劑; 降溫速率; 精莢; 白蝦; Cryopreservation; Cryoprotectant; Freezing rate; White shrimp; Spermatophore; |
語文 | 英文(English) |
中文摘要 | 本研究進行白蝦雄性配子之超低溫冷凍保存試驗,考慮各種可能產生影響之因子如適用的稀釋液、三種抗凍劑的種類及濃度、平衡時間、五種降溫速率、冷凍方法及三種解凍速率, 加以一一測試並比較。白蝦的精子不具游動性,其冷凍保存前後結果以eosin-nigrosin 染色法評估其活存率百分比。實驗結果顯示,白蝦精莢之最佳超低溫冷凍保存效果之稀釋液、降溫流程、抗凍劑種類及抗凍劑濃度為:以Ca-F saline作為稀釋液,濃度5% 之DMSO當作抗凍劑,於室溫平衡30 min後,以 -2 ℃/min之速率,將精莢降溫至 -80 ℃,維持2 min,之後再直接投入液態氮保存。樣品在30 ℃水浴中解凍,活存率為最高,達34.4 ± 3.4%。其次佳者為:以-1 ℃/min之速率降溫,活存率亦可達33.3 ± 3.9%,與前者並無顯著差異。在白蝦精莢長期保存試驗中,以濃度5% 之DMSO當作抗凍劑,室溫平衡30 min之分組經前處理後活存率為49.5 ± 8.3%;採用上述最佳流程保存1天後之活存率為44.3 ± 6.6%,其後隨著保存時間每10天觀察一次,直到第70天其活存率介於33 ~ 37% 之間,尚屬穩定,並無顯著差異 (P < 0.05)。經由本研究探討所得的結果,除了建立冷凍白蝦精莢目前最佳條件組合,並可提供作為日後白蝦雄性配子超低溫冷凍保存供實際應用時之參考。 |
英文摘要 | To study the cryopreservation of male gamete of white shrimp, Litopenaeus vannamei, various combinations of factors including proper extender, kind and concentration of three cryoprotectants, four equilibration times, five freezing rates and three thawing temperatures were examined. Practical cryopreservation of spermatophore in white shrimp was carried out. Due to the immobility of spermatozoa of white shrimp, the viability percentages of sperm before and after cryopreservation were assessed by staining the sperm cells with eosin-nigrosin. Best result was obtained in the subgroup in which shrimp spermatophores were treated with Ca-F saline (Calcium-free marine crustacean saline) at 25 oC, equilibrated in cryoprotectant of 5% DMSO for 30 min, frozen at rate of -2 oC/min until -80 oC, maintained for 2 min, and quenched in liquid nitrogen (LN2, -196 oC). The best thawing was found using water bath of 30 oC. The highest viability of sperm out of cryopreserved spermatophores based on this protocol was 34.4 ± 3.4%. Almost similar sperm viability (33.3 ± 3.9%) was observed in the subgroup cryopreserved at freezing rate of -1 oC/min. For long-term cryopreservation, equilibration in cryoprotectant of 5% DMSO for 30 min at ambient temperature of 25 oC resulted in 49.5 ± 8.3% viability in equilibrated sample, 44.3 ± 6.6% viability in one-day cryopreserved sample, and 33.0 to 37.0% in 10-, 20-, 30-, 40-, 50-, 60-,and 70-day cryopreserved samples. There was no significant difference among the last seven subgroups. This is the first report on the establishment of cryopreservation protocol for spermatophores of white shrimp based on the systematic combination of favorable factors. Further studies on the application could provide information for commercialization for both research and industrial use. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。