摘要
本实验从雨生红球藻中提取虾青素,添加到中华绒螯蟹(Eriocheir sinensis)配合饲料中,从抗氧化角度入手,在个体和细胞水平上研究虾青素对中华绒螯蟹的影响,以期全面评价虾青素在改善中华绒螯蟹的生理功能和提高中华绒螯蟹抵抗外界不良环境能力等方面的作用,进而推测虾青素的作用机理。
本文分别从虾青素投喂中华绒螯蟹的最适添加量的确定；虾青素的不同投喂方式对中华绒螯蟹的影响；虾青素对中华绒螯蟹抗环境胁迫能力的影响；虾青素对中华绒螯蟹的光保护作用；虾青素对中华绒螯蟹卵巢发育的影响五个方面进行探讨。
根据不同的实验目的所选取的评价指标有：生长生理指标(存活率、蜕皮频率、抗病菌侵染能力、Na+-K+-ATPase)；抗氧化指标：(CAT、MDA、总抗氧化能力)；血液免疫指标(血细胞计数、血淋巴吞噬百分比)；性腺发育指标(性腺指数、肝胰腺指数、卵母细胞直径、卵黄蛋白含量)；分子水平(DNA损伤)。
通过实验证明：虾青素作为免疫增强剂可以提高中华绒螯蟹的存活率和抗氧化能力p<0.05),其最适添加量为80 mg/kg,最佳投喂时间为4周；虾青素连续投喂时间过长可以引起免疫疲劳,间隔投喂可以消除虾青素引起的免疫疲劳,提高中华绒螯蟹免疫力,使中华绒螯蟹的生理机能更加稳定；添加虾青素的中华绒螯蟹抗环境胁迫能力和抗病菌侵染能力增强,能够有效的抵抗温度突变(15℃—28℃)、盐度突变(0—10‰)、致病菌(嗜水气单孢菌Aeromonas hydrophila浸浴感染,106ind.／mL)三种因子的胁迫：虾青素具有强抗氧化性,可作为潜在的光保护剂,使细胞免受UVA所造成的损伤；虾青素可以促进中华绒螯蟹雌蟹性腺发育,促进卵母细胞的成熟和卵黄蛋白的积累。
In this paper, the astaxanthin was extracted from Haematococcus pluvialis and added to the feedstuff of Eriocheir sinensis. The effect of astaxanthin upon E. sinensis was studied in terms of anti-oxidation at individual and cell levels so as to give an overall appraisal of astaxanthin used to improve E. sinensis physiological function and resistance to environmental stresses. Furthermore the mechanism of astaxanthin action could be predicted.
The effect of astaxanthin upon the growth of E. sinensis is discussed as below:
The optimum allowance of astaxanthin added to the feedstuff of E. sinensis.
The effect of different feeding ways of astaxanthin upon E. sinensis;
The effect of astaxanthin upon the resistance of E. sinensis to the environmental stresses;
Astaxanthin's anti-UVA function to E. Sinensis;
The effect of astaxanthin upon ovary development of E. Sinensis;
Evaluation factors were selected according to various experiment aims:growing and physiologic index (survival rate, moulting frequency, resistance to pathogenic bacteria, activity of Na+-K+-ATPase); anti-oxidation index (CAT, MDA, T-AOC); immunological index in hemolymph (Total haemocyte counts, phagocytic percentage of haemocyte); gonad development index (gonadosomatic index, hepatopancreas index, diameter of oocyte, the content of Vitellin); index in molecular level (DNA damage). Following aspects were proved through the experiments:
As an immunostimulant, Astaxanthin could increase the survival rate and anti-oxidation activity (P<0.05) of E. sinensisi. The optimum allowance of astaxanthin added to the feedstuff was 80mg/kg and the optimum-feeding span was 4 weeks.
Long-time feeding with astaxanthin consistently would cause immuno-fatigue in E. sinensis. Feeding in interval could solve this problem and improved immunity and made the physiological function more stable.
The endurance to the negative environmental factors and resistance to pathogenic bacteria could be enhanced in E. sinensisi fed with astaxanthin. The experiment also showed feeding with astaxanthin could improve endurance of E. sinensisi in acute change of temperature(15-28℃), salinity (0-10‰) and pathogenic Bacteria (106 ind./mL)
Astaxanthin that showed strong antioxidant ability could be used as photoprotectant, protecting cells from being damaged by UVA.
Astaxanthin could stimulate the ovarian development of E. sinensis and accumulate protein in oocyte and vitellin.
引文
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