【作者】 俞泉宇；

【导师】 李义；

【作者基本信息】 苏州大学 ， 水产养殖， 2013， 硕士

【摘要】 以毛竹笋为原料,采用超声辅助提取技术,研究了水料比、提取温度、提取时间、超声功率和乙醇浓度对竹笋多糖得率的影响,在此基础上应用响应面法对提取工艺进行优化。结果表明,竹笋多糖的最佳提取工艺为：水料比30：1、提取温度50℃、提取时间35min、乙醇浓度90%,在此条件下,多糖得率的理论值为2.944%,实际得率为2.938%；通过理化性质、紫外及红外光谱分析,证明提取物为多糖并推测可能是一种α-吡喃葡聚糖；苯酚-硫酸法测得多糖含量为65.3%。以Vc为对照,考察了0.1～10mg/mL的竹笋多糖对羟基自由基(·OH)、超氧阴离子(02-·)、过氧化氢(H1202)的清除效果及其脂质过氧化(LPO)抑制活性。结果显示,竹笋多糖对02-和H202的清除能力较强,高于同浓度的Vc；对清除·OH和抑制LPO的作用弱于Vc。由此表明,竹笋多糖具有较强的体外抗氧化活性。以注射等体积的灭菌生理盐水为对照,以1、5、10和20mg/kg体质重的竹笋多糖对中华绒螯蟹进行体腔注射,在注射后第1、4、7、10d采样,进行一系列免疫学相关指标的测定,并在注射多糖后的第4d,从每个处理组中取蟹6只,以1.2×107cfu/kg体质重的剂量体腔注射接种中华绒螯蟹致病性嗜水气单胞菌CL99920菌株,记录接种7d后各组蟹的累积死亡率。结果显示：注射适量的竹笋多糖能显著地增加中华绒螯蟹的血细胞总数(THC)和血细胞吞噬活性,显著地增强血清酚氧化酶(PO)、过氧化物酶(POD)和溶菌酶(LSZ)活性及肝胰腺一氧化氮合酶(NOS)活性,血清和肝胰腺超氧化物岐化酶(SOD)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)活性和总抗氧化能力(T-AOC)(P<0.05);对血细胞呼吸爆发活性(02-产量)没有显著影响(P<0.05)；在本试验条件下,竹笋多糖能明显增强中华绒螯蟹对嗜水气单胞菌的抗感染能力；从整体上看,以10mg/kg剂量的效果较佳。更多还原

【Abstract】 The water-soluble polysaccharide was extracted from the shoots of Phyllostachys edulis with ultrasonic wave. The response surface methodology was applied to determine the optimum technological conditions. The results indicated that the optimum extraction conditions were as follows:the water-material ratio was30:1, extraction temperature was50℃, extraction time was35min, ethanol concentration was90%. Under the conditions mentioned above, the predicted yield of bamboo shoots polysaccharide was2.944%, and the experimental yield was2.938%. The physical and chemical properties, ultraviolet spectroscopy and infrared spectroscopy showed that the extract was polysaccharide and presumably was a kind of α-pyran-glucan. The total sugar content of the polysaccharide was65.3%measured by phenol-sulfuric acid method.The scavenging effects on hydroxyl radicals, superoxide anion radicals and hydrogen peroxide and the inhibitory effect on lipid peroxidation of bamboo shoots polysaccharide on concentration0.1～10mg/mL were studied with ascorbic acid as a positive control. The results showed that bamboo shoots polysaccharide had significant scavenging effects on superoxide anion radicals and hydrogen peroxide which were higher than the same concentration of ascorbic acid. Otherwise, the scavenging effect on hydroxyl radicals and the inhibitory effect on lipid peroxidation were lower than that of the ascorbic acid. This study indicated that bamboo shoots polysaccharide had strong in vitro antioxidant activity.Crab were coelom injected at1,5,10and20mg/kg weight with bamboo shoots polysaccharide, with injected0.1mL sterile saline group as control. A series of immunology parameters were examined at1,4,7, and10days after administration of polysaccharide. Crab were challenged at1.2×107colony forming units (cfu)/kg weight with a virulent strain of Aeromonas hydrophila (CL99920) of E. sinensis at the second sampling time (4d), and mortalities were recorded over a7-day period. The results demonstrated that the total haemocyte count (THC), the haemocyte phagocytic activity, the phenoloxidase (PO), peroxidase (POD), lysozyme (LSZ) in serum, the nitrie oxide synthase (NOS) in hepatopancreas and the superoxide dismutase (SOD), acid phosphatase (ACP), alkaline phosphatase (ALP), total antioxidant capacity (T-AOC) in serum and hepatopancreas of E. sinensis were significant higher than those of the saline control group (P<0.05). However, no significant differences were observed in the respiratory burst (release of superoxide anion) in haemocyte of E. sinensis (P>0.05). Furthermore, the crabs administered with bamboo shoots polysaccharide were more resistant under the conditions of this experiment. On the whole, the crabs injected at10mg/kg weight with bamboo shoots polysaccharide had a stronger non-specific immune response and resistance to A. hydrophila.更多还原