载牛冠状病毒N蛋白壳聚糖微球的免疫效果评价

为了对壳聚糖分子作为佐剂的缓释效果进行免疫学评价,本研究合成了牛冠状病毒DB2毒株N蛋白基因的3'端第487位～1287位碱基,用大肠杆菌对其进行了高效表达,通过SDS-PAGE电泳分离、电洗脱纯化该牛冠状病毒N蛋白(BCV N).以戊二醛为交联剂,采用乳化交联的方法制备空白壳聚糖微球.利用吸附法制备载BCVN蛋白的壳聚糖微球,通过肌肉注射途径免疫BALB/c小鼠,用间接ELISA方法检测免疫鼠血清中的抗体水平,由此评价壳聚糖微球对BCVN的缓释作用和佐剂效应.结果表明,壳聚糖微球的形态较好、表面光滑、分散性较好,平均粒径为6.50±1.77μm,电势分布在36 mV,吸附BCV N蛋白的壳聚糖微球在体内所产生的免疫效果明显优于唯N蛋白组,说明载BCV N蛋白的壳聚糖微球在体内具有一定的缓释效果,使N蛋白在体内缓慢释放而长时间诱导抗体的产生,该研究结果为以壳聚糖微球作为疫苗佐剂积累了实验数据.     

稀土壳聚糖螯合盐对水产颗粒饲料性能的影响

研究了在饲料中添加稀土壳聚糖螯合盐（ＲＥＣＣ）对水产颗粒饲料性能的影响。根据鲫（Ｃａｒａｓｓｉｕｓａｕｒａｔｕｓ）鱼苗的营养需求配制４种类型的ＲＥＣＣ试验日粮，即１号饲料（０．００％）、２号饲料（０．０８％）、３号饲料（０．１６％）和４号饲料（０２４％）。测定４种饲料的粉化率和溶失率。结果显示，添加稀土壳聚糖螯合盐后可以降低饲料的 粉化率，饲料中添加０．１６％的稀土壳聚糖螯合盐效果尤其明显（Ｐ＜０．０１）；在饲料中添加ＲＥＣＣ可以显著降低饲料的溶失率（Ｐ＜０．０５），并可以提高和改善水产颗粒饲料的性能。     

壳聚糖的生产与应用

近年来,壳聚糖及其衍生物的开发应用受到国内外的重视,越来越多的产品从实验室走向市场。介绍了壳聚糖及其衍生物的制备方法及其在工业、农业、医药、环保、生物学等领域的应用。     

壳聚糖对丰都红心柚保鲜效果的影响

采用H202氧化降解法制备相对分子质量(Mw)为1 500～2000的低分子质量壳聚糖.先用有益微生物制剂100液浸果2 min,再用浓度为1.5%的低分子质量壳聚糖涂膜保鲜丰都红心柚.试验结果表明,在常温和普通民房贮藏条件下贮藏100天,其烂果率为9.2%、干疤率为10.3%,防腐保鲜效果优于化学防腐保鲜剂.     

壳聚糖对草莓冷藏和货架期间品质的影响

以"达赛莱克特"草莓为试材,研究1%壳聚糖处理的草莓果实在0℃低温贮藏7 d后20℃常温货架6 d期间的品质变化规律。结果表明:与对照(0.1%醋酸溶液处理)相比,壳聚糖处理降低了低温贮藏后期和货架期草莓果实的呼吸速率、花青素、类黄酮及酚类物质的含量,保持较高的果实硬度,显著减少了货架期草莓果实腐烂和霉菌的发生,对果实失重率和可溶性固形物含量无显著影响。表明1%壳聚糖处理可有效延缓草莓的采后衰老进程,延长果实的货架期。     

壳聚糖保鲜液对唐菖蒲切花保鲜的影响

试验采用0.25‰,0.5‰,0.75‰,1‰,1.25‰的壳聚糖保鲜液处理唐菖蒲切花,观察切花寿命和开花率,旨在探索壳聚糖保鲜液处理切花的效果。试验结果表明,1‰壳聚糖保鲜液处理后的鲜切花,寿命最长,开花率最高。     

胰蛋白酶对壳聚糖的降解研究

用粘度法和吸光度法，研究胰蛋白酶非专一性降解壳聚糖过程中温度、PH值、反应时间、酶浓度、底物浓度。金属离子对胰蛋白酶降解壳聚糖反应的影响，确定了以壳聚糖为底物的胰蛋白酶的一些催化特性：最适温度为30℃，最适PH值为5.0,10-180min内酶反应速度恒定，酶浓度在0.1-0.5g/L的范围内，酶反应速度与酶浓度呈线性关系，米氏常数Km为9.54g/L。     

新饲料添加剂N，O-羧甲基壳聚糖的急性毒性研究

【目的】研究N,O-羧甲基壳聚糖对鸡的急性毒性,评价该药的安全性;【方法】以最大浓度的N,O-羧甲基壳聚糖(10.5%),按20ml/(kg·bw)(2.1g/(kg·bw)/次)剂量24h内给鸡灌服;【结果】给药后连续观察7天,各组试验鸡全部存活,临床、剖检均未见到异常变化,测不出LD50,根据新药审批办法中关于急性毒性的要求,进行最大耐受量的测定。鸡灌服给予N,O-羧甲基壳聚糖的最大耐受量为10.5g/(kg·bw),相当于临床拟用量的3500倍;【结论】N,O-羧甲基壳聚糖毒副作用很小,临床可以安全应用。     

Chitosan nanoparticles enhance developmental competence of in vitro-matured porcine oocytes

Oxidative stress is inevitable as it is derived from the handling, culturing, inherent metabolic activities and medium supplementation of embryos. This study was performed to investigate the protective effect of chitosan nanoparticles (CNPs) on oxidative damage in porcine oocytes. For this purpose, cumulus–oocyte complexes (COCs) derived from porcine slaughterhouse ovaries were exposed to different concentrations of CNPs (0, 10, 25 and 50 µg/ml) during in vitro maturation (IVM). Oocytes treated with 25 µg/ml CNPs showed significantly higher levels of GSH, along with a significant reduction in ROS levels compared to control, CNPs10 and CNPs50 groups. In parthenogenetic embryo production, the maturation rate was significantly higher in the CNPs25 group than that in the control and all other treated groups. In addition, when compared to the CNPs50 and control groups, CNPs25-treated oocytes showed significantly higher cleavage and blastocyst development rates. The highest concentration of CNPs reduced the total cell number and ratio of ICM: TE cells in parthenogenetic embryos, suggesting that there is a threshold where benefits are lost if exceeded. In cloned embryos, the CNPs25 group, as compared to all other treated groups, showed significantly higher maturation and cleavage rates. Furthermore, the blastocyst development rate in the CNPs25-treated group was significantly higher than that in the CNPs50-treated group, as was the total cell number. Moreover, we found that cloned embryos derived from the CNPs25-treated group showed significantly higher expression levels of Pou5f1, Dppa2, and Ndp52il genes, compared with those of the control and other treated groups. Our results demonstrated that 25 µg/ml CNPs treatment during IVM improves the developmental competence of porcine oocytes by reducing oxidative stress.     

NaCl plus chitosan as a dietary salt to prevent the development of hypertension in spontaneously hypertensive rats

The effect of NaCl plus 3% chitosan on the systolic blood pressure of spontaneously hypertensive rats (SHR) were evaluated and compared with NaCl plus KCl (NaCl, 49.36% + KCl 49.36%) and chitosan or NaCl treatment alone. In SHR, administration of NaCl plus chitosan (44 mM Na/day) for two months significantly decreased the systolic blood pressure greater than of NaCl plus KCl and NaCl alone. NaCl plus chitosan resulted, though not statistically significant, in decreased urinary Na⁺ excretion and decreased blood urea nitrogen levels. Urinary creatinine of NaCl plus chitosan was slightly decreased compared to 3 treated groups. Serum electrolytes levels, however, remained unchanged. The combination of NaCl and chitosan may be superior to the conventional use of NaCl plus KCl or NaCl alone in the prevention of hypertension. Even though these supplementary diets have demonstrated potential anti-hypertensive effects in the experimental animal model, further research is needed before any recommendations can be made.