淡水粗糙沼蝦繁殖生物學之研究

Studies on the Reproductive Biology in the Freshwater Prawn Macrobrachium asperulum (Decapoda: Palaemonidae)

黃楷翔(Kai-Hsiang Huang)

指導教授 ： 陳弘成

國立臺灣大學/生命科學院/漁業科學研究所/博士(2010年)

https://doi.org/10.6342/NTU.2010.03117

摘要

粗糙沼蝦(Macrobrachium asperulum)為臺灣常見之淡水蝦，其經濟價值高且具量產開發之潛力。然而，目前關於粗糙沼蝦繁殖相關之研究仍然不多亦不夠詳細。因此，本試驗擬訂將野外採樣與收集之粗糙沼蝦，予以外部型態之調查、抱卵率與性腺成熟指數(GSI)之測量與換算、類卵黃前質素蛋白含量之檢測與卵巢組織切片之觀察等。依據檢測結果，訂定母蝦繁殖週期、生殖細胞、卵巢與胚胎發育之型態與特徵。另外，內分泌干擾物質雙酚A(bisphenol A)對沼蝦生殖毒性以及血清素(serotonin)於母蝦性腺催熟效用部份，本試驗亦予以試驗與評估。
每年三月至九月為粗糙沼蝦之繁殖季節。雨量與光週期增加可能是啟動粗糙沼蝦卵巢發育之重要環境因子，而水溫增加則可能與母蝦性線成熟有關。母蝦生殖細胞大致分為卵原細胞、初級卵母細胞、次級卵母細胞與成熟卵母細胞等。其中，次級卵母細胞根據其外部型態，再細分為五類。粗糙沼蝦為一年多生之淡水蝦，且其卵巢發育分為增殖期、卵黃生合成前期、內源性卵黃生合成期、外源性卵黃生合成期、成熟期與排卵期等。此六個性腺發育階段由上述八種生殖細胞所組成，每個階段生殖細胞組成比例皆不同。另外，卵黃生合成階段，母蝦體內類卵黃前質素蛋白含量逐漸增加直至卵巢發育成熟，接著隨著母蝦產卵，此蛋白質含量才顯著地減少。粗糙沼蝦抱卵孵化時間為39.17 ± 4.36日、抱卵數為86.40 ± 17.40粒。隨著胚胎發育，卵粒顏色變化依序為翠綠色、橄欖綠色、黃綠色與淺黃綠色。卵受精20天，卵粒可觀察到幼蝦複眼之構造。卵孵化前三天，卵粒可清楚發現幼蝦外觀之輪廓。另外，卵黃前質素為母蝦特有之蛋白質，本試驗證實內分泌干擾物質雙酚A刺激公蝦生合成母蝦特有之類卵黃前質素蛋白。除此之外，本試驗發現5 μg/g bw 血清素顯著地刺激母蝦性腺發育。單邊眼柄切除處理方式可能無法顯著地刺激沼蝦卵巢發育，而致死率較高之雙邊眼柄切除技術則能顯著地促進母蝦性腺發育。
本試驗證實公蝦體內類卵黃前質素蛋白含量為一項有效用於監控環境類雌性素污染之生物指標，且粗糙沼蝦亦適合做為指標生物。另外，本試驗胚胎發育結果相信將有助於淡水蝦幼苗之生產與管理，進而減少卵孵化場之工作天數，並降低生產大量幼苗所耗費之成本。本試驗不僅使淡水甲殼類動物繁殖生物學更趨於完整，且為將來粗糙沼蝦養殖量產或保育復育皆提供重要之參考依據。

關鍵字

雙酚A ； 胚胎發育 ； 粗糙沼蝦 ； 繁殖生物學 ； 血清素 ； 卵黃生合成作用

並列摘要

Reproductive biology, especially, the reproductive physiology, in Macrobrachium asperulum was investigated in the present study. The breeding period of M. asperulum is from March to September. Developing ovaries and germ cells were characterized throughout the entire ovarian cycle. In this study, differentiating germ cells were classified mainly into oogonia, primary oocytes, secondary oocytes, and mature oocytes. Secondary oocytes were further divided into five subtypes. Summarizing the above examinations and based on the relative frequency of germ cell types, six ovarian stages were identified, and M. asperulum is proposed as being a multiple spawning species. Moreover, a gradual increase in levels of the hemolymph, hepatopancreas and ovary vitellogenin-like protein were recorded during the stages I-IV. These values peaked in stage V (mature) and then declined in stage VI (spawning). After ovulation, the eggs-carrying period was about 39.17 ± 4.36 days, and the clutch size was about 86.40 ± 17.40. Egg morphology was also characterized throughout the embryogenesis.
A world wide-distributed chemical bisphenol A, an endocrine disrupter, found in the effluents of industrial and municipal sewage treatment plants, can be able to elicit the induction of biomarker vitellogenin-like protein in male M. asperulum. In addition, prawns were perceived sensitive to the estrogenic compounds.
The administration of 5 μg/g bw serotonin injection can be able to trigger ovarian development. In addition, compared to unilaterally eyetalk ablated females, bilateral eyestalk ablation had an obvious stimulatory effect on the development of ovaries, although its high mortality was recorded.
Male M. asperulum as a biomonitoring species could be considered and proposed, and levels of vitellogenin-like protein in males can be a useful biomarker of estrogenic contaminations. In addition, the biology described in present study may be beneficial in reducing hatchery man-days and the cost of production of prawn seeds. Moreover, the present study will not only integrate the field of the reproductive biology of decapod crustaceans, but also provide significant information for the future aquaculture and/or conservation of M. asperulum.

並列關鍵字

bisphenol A ； embryonic development ； Macrobrachium asperulum ； reproductive biology ； serotonin ； vitellogenesis

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