【作者】 梁超；

【导师】 张文兵；

【作者基本信息】 中国海洋大学 ， 水生生物学， 2011， 硕士

【摘要】 本文以我国重要的海水养殖动物——刺参(Apostichopus japonicas Selenka)为研究对象,探讨饲料中添加二甲酸钾、低聚木糖和菊粉对刺参生长、免疫以及肠道菌群的影响。主要研究内容和结果如下:1.利用单因素实验设计,以初始体重为(2.13±0.02)g的刺参(Apostichopus japonicas Selenka)为研究对象,分别在基础饲料中添加0%、0.2%、0.4%、0.6%、0.8%、1.0%和1.2%的二甲酸钾配制出7种实验饲料,在海水循环系统中养殖8周,探讨饲料中添加二甲酸钾对刺身生长、免疫和肠道菌群的影响。结果表明:二甲酸钾对刺参的成活率没有影响,各处理组均为100%;对生长有显著的促进作用,当添加量为1.0%和1.2%的时候,刺参的特定生长率(SGR)要显著高于对照组(P<0.05)。饲料中添加1.0%或1.2%的二甲酸钾,显著提高了刺参体腔细胞的吞噬活性及活性氧产量(P<0.05)。二甲酸钾对超氧化物歧化酶(SOD)活性及一氧化氮合酶(NOS)活性没有显著影响。当饲料中添加0.4%、0.6%、0.8%、1.0%或1.2%的二甲酸钾时,刺参肠道弧菌数显著低于对照组(P<0.05),但对总菌数没有显著影响(P>0.05)。通过注射刺参腐皮综合症致病菌灿烂弧菌(V.splendidus )进行刺参攻毒实验,结果表明对照组和0.2%添加组刺参累积死亡率为46.67%、38.33%,显著高于0.4%、0.6%、0.8%、1.0%、1.2%二甲酸钾添加组(P<0.05)。2.利用单因素实验设计,以初始体重为(2.14±0.02)g的刺参(Apostichopus japonicas Selenka)为研究对象,分别在基础饲料中添加0%、0.02%、0.04%、0.06%、0.08%、0.10%、0.12%的低聚木糖配制出7种实验饲料,在海水循环系统中养殖8周,探讨饲料中添加低聚木糖对刺身生长、免疫和肠道菌群以及抗病力的影响。结果表明:低聚木糖对于刺参的成活率没有影响,各处理组均为100%(P>0.05)。对刺参的生长有显著的促进作用,当添加量为0.06%、0.08%、0.10%、0.12%的时候,刺参的特定生长率(SGR)要显著高于对照组(P<0.05)。与对照组相比,当饲料中添加0.1%、0.12%的低聚木糖,显著提高了刺参体腔细胞的超氧化物歧化酶(SOD)活性(P<0.05),饲料中添加0.08%、0.10%、0.12%的低聚木糖显著提高了刺参体腔细胞一氧化氮合酶(NOS)活性,高于对照组及其他处理组(P<0.05)。低聚木糖对细胞吞噬活性及细胞内活性氧产量没有显著影响(P>0.05),但有上升趋势。当饲料中添加0.10%、0.12%低聚木糖时,刺参肠道总菌数显著高于对照组(P<0.05),但添加低聚木糖对刺参肠道弧菌数没有显著影响。通过注射刺参腐皮综合症致病菌灿烂弧菌(V.splendidus )进行刺参攻毒实验,攻毒14d后发现,饲料中添加不同梯度的低聚木糖对于刺参抗病能力没有显著影响(P>0.05)。但随着低聚木糖添加量的提高,刺参感染灿烂弧菌后的死亡率有降低的趋势,在0.10%添加组时刺参死亡率最低(30.2%)。3.利用单因素实验设计,以初始体重为(2.14±0.02)g的刺参(Apostichopus japonicas Selenka)为研究对象,分别在基础饲料中添加0%、0.1%、0.3%、0.5%、0.7%、1.0%菊粉配制出6种实验饲料,在海水循环系统中养殖8周,探讨饲料中添加菊粉对刺身生长、免疫和肠道菌群以及抗病力的影响。结果表明:菊粉对于刺参的成活率没有影响,各处理组均为100%(P>0.05)。对刺参的生长有显著的促进作用,当添加量为0.7%、1.0%时刺参的特定生长率(SGR)(P<0.05)最高。饲料中添加菊粉可以显著提高刺参体腔细胞的超氧化物歧化酶(SOD)活性和一氧化氮合酶(NOS)活性(P<0.05),0.7%和1.0%处理组与对照组差异显著。刺参细胞吞噬活性及细胞内活性氧产量随着饲料中菊粉的添加量呈上升趋势,在1.0%添加量时效果最好。当饲料中添加0.7%的菊粉时,刺参肠道总菌数显著高于对照组(P<0.05)。当饲料中添加0.5%的菊粉时,刺参肠道弧菌数显著低于对照组(P<0.05),其余处理组与对照组差异不显著(P>0.05)。通过注射刺参腐皮综合症致病菌灿烂弧菌(V.splendidus )进行刺参攻毒实验,攻毒后14d后发现,随着菊粉添加量的提高,刺参感染灿烂弧菌后的死亡率有降低的趋势,在1.0%添加组时刺参死亡率最低(27.78%)。更多还原

【Abstract】 Three feeding trials were conducted in indoor circulating system to investigate the effects of dietary potassium diformate, xylo-oligosaccharides and inulin on growth, immunity and disease resistance of sea cucumber (Apostichopus japonicus Selenka). The results were summarized as follows:1.A 8-week feeding experiment was conducted to investigate the effects of dietary potassium diformate on growth, immune responses, intestinal mircroflora and disease resistance of sea cucumber Apostichopus japonicas Selenka (mean initial body weight 2.13±0.03g). Potassium diformate was supplemented into the basal diet to formulate seven isonitrogenous and isoenergetic practical diets containing 0.00% (the control diet), 0.2%, 0.4%, 0.6%, 0.8%, 1.0% and 1.2% potassium diformate of dry weight, respectively. The results showed that there was no mortality observed during the feeding trial. The specific growth rate (SGR) of sea cucumber was significantly enhanced by the supplementation of potassium diformate in diets. Sea cucumber fed diets with 1.0% and 1.2% potassium diformate showed significantly higher SGR compared to the control group (P<0.05). The immune function assay showed that dietary supplementation of 1.0% and 1.2% potassium diformate significantly increased the coelomocytes phagocytosis and the active oxygen species production compared to other treatments (P<0.05). No significant difference among dietary treatments were observed in superoxide dismutase and nitric oxide synthase (P>0.05). The Vibiro spp. counts in sea cucumber fed diets with 0.4%, 0.6%, 0.8%, 1.0% and 1.2% potassium diformate were significantly lower than the control group (P<0.05). However, the total viable bacterial counts were independent of dietary treatments. The challenge experiment with V. splendidus showed that 14-day cumulative mortality in sea cucumber fed the control diet (46.67%) and 0.2% potassium diformate (38.33%) was significantly higher than those in sea cucumber fed diets with 0.4%, 0.6%, 0.8%, 1.0% and 1.2% potassium diformate (P<0.05).2. A 8-week feeding experiment was conducted to investigate the effects of dietary xylo-oligosaccharides on growth, immune responses, intestinal mircroflora and disease resistance of sea cucumber Apostichopus japonicas Selenka (mean initial body weight 2.13±0.03g). Xylo-oligosaccharides was supplemented into the basal diet to formulate seven isonitrogenous and isoenergetic practical diets containing 0.00% (the control diet), 0.02%, 0.04%, 0.06%, 0.08%, 0.10% and 0.12% xylo-oligosaccharides of dry weight, respectively. The results showed that there was no mortality observed during the feeding trial. The specific growth rate (SGR) of sea cucumber was significantly enhanced by the supplementation of xylo-oligosaccharides in diets. Sea cucumber fed diets with 0.06%,0.08%, 0.10% and 0.12% xylo-oligosaccharides showed significantly higher SGR compared to the control group (P<0.05). The immune function assay showed that dietary supplementation of 0.10% and 0.12% xylo-oligosaccharides significantly increased the superoxide dismutase activity compared to the control and the dietary supplementation of 0.10% and 0.12% xylo-oligosaccharides significantly increased the nitric oxide synthase activity compared to other treatments (P<0.05). Increasing trends were observed in the coelomocytes phagocytosis and the active oxygen species production though there were no significant differences among dietary treatments in the coelomocytes phagocytosis and the active oxygen species production (P>0.05).The total viable bacterial counts in sea cucumber fed diets with 0.10% and 1.2% xylo-oligosaccharides were significantly higher than the control group (P<0.05). However, the Vibiro spp. counts were independent of dietary treatments. The challenge experiment with V. splendidus showed that there were no significant differences in 14-day cumulative mortality among dietary treatments (P>0.05). But with the increase of dietary xylo-oligosaccharides levels, 14-day cumulative mortality of sea cucumber decreased. The lowest 14-day cumulative mortality was found in sea cucumber fed 0.10% xylo-oligosaccharides. 3.A 8-week feeding experiment was conducted to investigate the effects of dietary inulin on growth, immune responses, intestinal mircroflora and disease resistance of sea cucumber Apostichopus japonicas Selenka (mean initial body weight 2.13±0.03g). Inulin was supplemented into the basal diet to formulate seven isonitrogenous and isoenergetic practical diets containing 0.00% (the control diet), 0.1%, 0.3%, 0.5%, 0.7%, 1.0% inulin of dry weight, respectively. The results showed that there was no mortality observed during the feeding trial. The specific growth rate (SGR) of sea cucumber was significantly enhanced by the dietary supplementation of inulin and sea cucumber fed 0.7% and 1.0% inulin showed the highest SGR. The immune function assay showed that dietary inulin supplementation increased the superoxide dismutase activity (P<0.05) and the nitric oxide synthase activity compared to other treatments (P<0.05). Sea cucumber fed diets with 0.7% and 1.0% inulin showed increased activities of both these two enzymes compared to the control group. Increasing trends were observed in the coelomocytes phagocytosis and the active oxygen species production with the increase of dietary inulin and the 1% inulin supplementation group showed the highest values. The total viable bacterial counts in sea cucumber fed diets with 0.7% inulin were significantly higher than the control group (P<0.05) and the Vibiro spp. counts in sea cucumber fed diets with 0.5% inulin were significantly lower than the control group (P<0.05). However, there were no differences among the other treatments. The challenge experiment with V. splendidus showed that 14-day cumulative mortality decreased with the increase of dietary inulin and 14-day cumulative mortality in sea cucumber fed the diet with 1.0% inulin was significantly lower than the other treatments.更多还原