油茶皂苷的制备及提纯工艺研究

为研究油茶皂苷的制备及提纯工艺,为中试化生产提供科学的技术支撑,通过正交试验得到最佳工艺为乙醇体积分数75%,料液比1∶2,提取时间120 min,提取温度60℃,得到纯度为55.65%的油茶皂苷粗品,得率16.12%。油茶皂苷粗品配置成质量分数3%水溶液,加入10%体积的1%壳聚糖絮凝剂絮,经凝沉、淀除去杂质,澄清液经AB-8型大孔树脂吸附90 min,用质量分数2%氢氧化钠溶液洗脱杂质,再用体积分数70%乙醇溶液洗脱油茶皂苷,将洗脱液旋转蒸发,除去乙醇并烘干,即得到纯度85.36%的油茶皂苷。     

1-MCP和壳聚糖处理保持红枣贮藏品质及其机理研究

以新疆灰枣为试材，研究纸片型1-甲基环丙烯（1-methylcyclopropene，1-MCP）与壳聚糖（chitosan，COS）处理在（0±1）℃条件下对灰枣品质及相关酶活性的影响。结果表明：至贮藏90 d时，与对照（CK）相比，1-MCP＋COS的腐烂率、失质量率分别降低53.4%、51.2%，可溶性固形物（total soluble solid, TSS）含量高出18.6%。抗氧化酶系统中的超氧化物歧化酶（superoxide dismutase, SOD）峰值水平提高58.2%，过氧化氢酶（catalase, CAT）峰值高出34.8%；贮藏结束时过氧化物酶（peroxidase, POD）活性是CK的1.98倍，抗坏血酸过氧化物酶（ascorbate peroxidase, APX）活性高出51.7%，差异显著（P＜0.05）。1-MCP、COS及1-MCP+COS处理均可不同程度地降低果实腐烂率、失质量率，保持较高的果实硬度和TSS含量，推迟呼吸高峰和乙烯释放速率高峰的到来并抑制丙二醛（malondialdehyde, MDA）的积累，抑制脂氧合酶（lipoxygenase, LOX）和多酚氧化酶（polyphenol oxidase, PPO）活性，显著提高抗氧化酶系统活性。其中1.50 μL/L纸片型1-MCP与1% COS复合处理对保持果实贮藏品质、延缓果实衰老、延长货架期最有利。     

高光谱成像快速检测壳聚糖涂膜草莓可溶性固形物

为了对壳聚糖涂膜草莓可溶性固形物含量(soluble solids content, SSC)进行快速检测,该文采用高光谱成像仪(400～1 000 nm)对0,0.5%,1%浓度的壳聚糖(chitosan, CTS)涂膜草莓分别储藏1,2,4 d后进行成像,并测量样本SSC。通过分析SSC发现,0.5%和1%壳聚糖涂膜草莓,其SSC随着储藏天数的增加均高于0浓度壳聚糖涂膜草莓,说明了0.5%和1%壳聚糖涂层抑制了草莓中SSC的降低,能够延长草莓的新鲜口味。随后采用蒙特卡罗-偏最小二乘法(monte carlo-partial least squares, MCPLS)对异常样本进行剔除。对剔除异常样本后的光谱数据进行不同预处理,以确定最优的预处理方法。为提高运行速度和降低数据维数,采用竞争性自适应权重取样法(competitive adaptive reweighted sampling, CARS)和连续投影算法(successive projections algorithm, SPA)进行特征波段选择。最后,采用偏最小二乘回归(partial least square regression, PLSR)和支持向量回归(support vector regression, SVR)法建立回归模型。最终结果表明:SPA-SVR模型效果最佳,0浓度的壳聚糖涂膜的草莓,建模集精度Rc2为0.865,预测集精度Rv2为0.835;0.5%浓度的壳聚糖涂膜的草莓,建模集精度Rc2为0.808,预测集精度Rv2为0.799;1%浓度的壳聚糖涂膜的草莓,建模集精度Rc2为0.834,预测集精度Rv2为0.875。对储藏第4天的部分样本图像进行主成分分析(principal component analysis, PCA),结果显示除第二主成分图像(PC2)中有部分噪声影响外,PC1和PC3均能完整反映草莓信息,且PC3图像明显呈现出不同浓度壳聚糖涂膜草莓的褐变程度,说明不同浓度的壳聚糖涂膜也会对草莓货架期产生不同影响。综上说明利用高光谱成像技术可以实现壳聚糖涂膜草莓SSC快速检测,有效指导草莓保鲜处理。     

甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物纳米微球作为口蹄疫病毒核酸疫苗递送载体的特性评价

旨在对制备的甘露糖修饰的壳聚糖聚乳酸-羟基乙酸共聚物[poly （D，L-lactide-co-glycolide），PLGA]纳米微球作为口蹄疫病毒（foot-and-mouth disease virus，FMDV）核酸疫苗递送载体进行评价。采用西佛碱反应和元素分析制备具有一定取代度的甘露糖修饰的壳聚糖衍生物（mannose modified chitosan，MCS），然后，经双重乳化挥发法制备得到甘露糖修饰的壳聚糖PLGA纳米微球（MCS-PLGA-NPs）。采用纳米粒径仪检测MCS-PLGA-NPs粒径分布和表面电势（zeta）、扫描电镜考察其形态、琼脂糖凝胶电泳观察其对质粒的吸附和吸附质粒后抵抗核酸酶降解能力、CCK-8法检测MCS-PLGA-NPs的细胞毒性、激光共聚焦观察巨噬细胞对MCS-PLGA-NPs-质粒DNA复合物的摄取、荧光显微镜和Western blot验证MCS-PLGA-NPs加载质粒DNA在细胞中的表达。元素分析结果表明，成功制备了取代度为5%~10%的MCS。纳米粒径测定和扫描电镜结果表明，MCS-PLGA-NPs的zeta为正值、粒径分布均匀且形态规则呈球形。琼脂糖凝胶电泳结果显示，MCS-PLGA-NPs吸附质粒的能力随着其质量的增加而增强并且可以在一定程度上抵抗核酸酶降解质粒DNA。在细胞毒性试验中，不同浓度的MCS-PLGA-NPs与RAW264.7细胞共孵育24 h后，细胞存活率仍在85%以上。在细胞摄取试验中，用激光共聚焦显微镜可以明显观察到质粒DNA结合到纳米微球表面被RAW264.7细胞摄取。荧光显微镜和Western blot试验证明MCS-PLGA-NPs加载质粒DNA可以在细胞中进行表达。综上表明，本研究成功制备了MCS以及具有递送核酸疫苗能力的MCS-PLGA-NPs，为FMDV核酸疫苗的递送研究提供了新的方向和见解，也为该递送载体携带特定抗原靶向抗原递呈细胞表面甘露糖受体以及应用于动物免疫的研究奠定基础。     

Assessment of the Antibacterial Effect of Chitosan Coated with Heracleum persicum Oil on Rainbow Trout Quality

Today, due to the rapid spoilage of fish, the use of natural preservatives is a priority over those of synthetic varieties. Also, the natural antimicrobial effects of essential oils can help to increase shelf life. There are very few studies concerning the use of essential oils in this regard. In the present study, the antibacterial effect of chitosan coated with Heracleum persicum oil was investigated on the quality of rainbow trout. The control and the coated fish samples were analyzed periodically by generalized estimating equation (GEE) for total viable count (TVC), psychrophilic bacteria, total volatile base nitrogen (TVB-N), polyunsaturated fatty acids (PUFA), texture, taste, odor, and overall acceptance parameters. The results showed that by increasing the amount of chitosan, TVC, psychrophilic bacteria, and TVB-N decreased while PUFA, texture, taste, odor, and overall acceptance parameters increased.     

不同贮藏温度和保鲜处理对黄秋葵贮藏品质及生理的影响

为探讨采后不同贮藏温度对黄秋葵贮藏品质的影响以及不同保鲜剂对黄秋葵抗氧化酶活性的影响，以台湾“五福”黄秋葵为试材，分别置于低温（（4±1）℃、（9±1）℃、（14±1）℃）和常温（（20±1）℃）条件下贮藏，研究温度对其贮藏品质的影响；采用1.0%壳聚糖、5.0 mmol/L水杨酸、0.5片安喜布、5%O2+8%CO2处理黄秋葵，研究保鲜处理对其抗氧化酶活力的影响。结果表明：（9±1）℃贮藏组的保鲜效果最好，显著改善了贮藏期间黄秋葵的失重率、硬度、叶绿素含量、VC含量、L值、呼吸强度、乙烯释放量、MDA含量、细胞膜相对电导率；4种保鲜处理均能显著提高黄秋葵的SOD、POD、CAT活性，并维持其活性在相对较高的水平；同时，显著抑制黄秋葵果实MDA含量的升高。通过研究发现，壳聚糖涂膜液处理组的效果最佳，水杨酸处理组次之并优于1-MCP处理组，气调处理组的效果相对较差。     

日粮添加壳聚糖对鹅生长性能、血脂及脂代谢相关基因表达的影响

为研究日粮中添加不同剂量的壳聚糖对肉仔鹅生长性能、血脂成分、肝脏脂代谢关键酶及其mRNA表达的影响,探讨壳聚糖调控鹅脂质代谢的机理。选取14日龄扬州鹅公仔鹅360只,随机分为5组各6个重复,每个重复12只。即基础饲粮组(对照组,C),基础饲粮+250 mg/kg(Ⅰ),基础饲粮+500 mg/kg(Ⅱ)、基础饲粮+1 000mg/kg(Ⅲ)及基础饲粮+2 000mg/kg(Ⅳ)壳聚糖组,试验期56d。结果表明:1)饲粮中添加250和500mg/kg的壳聚糖均能显著提高试验鹅日采食量(ADFI)、日增重(ADG)及70日龄体重(P0.05);2)在250~1 000mg/kg范围内,和对照组相比,试验组血清总胆固醇(TC)及低密度脂蛋白(LDL)浓度显著降低(P0.05);不同处理组间血清甘油三酯(TG)、高密度脂蛋白(HDL)差异不显著(P0.05);3)饲粮中添加250和500mg/kg的壳聚糖能够显著降低肝脏乙酰辅酶A羧化酶(ACC)活性,并抑制ACC和脂肪酸合成酶(FAS)mRNA的表达(P0.05)。试验鹅肝脏过氧化物酶体增殖物激活受体α(PPAR-α)mRNA表达水平随着壳聚糖水平提高,显著上调;4)在250~2 000mg/kg范围内,壳聚糖添加量不影响试验鹅肝脏FAS及LPL活性(P0.05)。但500和1 000mg/kg壳聚糖能显著降低其肝脏HL活性。综上所述,壳聚糖能够通过影响肝脏脂肪代谢酶活性及脂代谢相关基因的表达,调控试验鹅血脂代谢,提高其生长性能,添加量以250~500mg/kg效果最佳。     

Influence of nanochitosan/zeolite composite on growth performance,digestive enzymes and serum biochemical parameters in rainbow trout (Oncorhynchus mykiss)

Protective effects of nanochitosan/zeolite composite besides zeolite and chitosan/zeolite composite on rainbow trout growth, digestive enzyme activities and biochemical parameters were studied. Nanochitosan/zeolite hybrid composites with three different ratio of nanochitosan:zeolite (35:100, 3.5:100 and 0.35:100) were synthesized and analysed by X‐ray powder diffraction (XRD), thermal gravim (TG) analysis and scanning electron microscope (SEM) methods. Prepared composites as well as zeolite and control diets were fed to rainbow trout (50 g) for a period of 60 days. The addition of treated diets significantly improved growth performance compared to the control diet. Supplemental zeolite could only enhance the amylase activity in fish intestine, whereas other treatment groups could increase the pepsin activity besides intestinal alkaline phosphatase, trypsin and amylase activities. No differences were observed for intestinal acid phosphatase and lipase activities among the experimental diets. Meanwhile, serum total antioxidant activity and lipid peroxidation product, indicated by malondialdehyde (MDA), significantly increased and decreased, respectively, with some doses of administration, indicating the elevated antioxidant status in treatment groups. Serum‐specific marker enzymes, namely aspartate aminotransferase (AST) and alkaline phosphatase (ALP), were not affected in all groups. Meanwhile serum total protein in most treatment groups was significantly higher than the control group. Results showed that synthesized composites especially nanochitosan/zeolite composite at 5 g/kg had potential to enhance growth performance, digestive enzyme activities and some biochemical parameters in rainbow trout.     

Effect of dietary chitosan oligosaccharide complex with Ce (IV) on growth,immunity and disease resistance against Vibrio splendidus of sea cucumber,Apostichopus japonicas

An 8‐week feeding trial was conducted to investigate the effects of dietary chitosan oligosaccharide complex with cerium (Ce IV) (COS‐Ce) on growth performance, nonspecific immunity and disease resistance of sea cucumber, Apostichopus japonicas. Five isonitrogenous (18.6%) and isolipidic (1.1%) practical diets were formulated with graded level of COS‐Ce (0, 150, 300, 600 and 1200 mg kg^?1 dry feed), which were named as COS‐Ce/0, COS‐Ce/150, COS‐Ce/300, COS‐Ce/600, COS‐Ce/1200 respectively. Each diet was allocated to four replicates of sea cucumbers (Initial weight: 6.72 ± 0.02 g). Sea cucumbers were fed to apparent satiation once daily (19:00 hours) for 56 days. During the experiment, water temperature was kept at 16 ± 0.5°C, pH 7.8–8.2, dissolved oxygen beyond 5 mg L^?1, ammonia nitrogen below 0.5 mg L^?1 and salinity from 30‰ to 31‰. Results showed that the specific growth rate of sea cucumbers was significantly higher in COS‐Ce/600 than that in other four treatments. Activities of phagocytosis, respiratory burst, acid phosphatase and alkaline phosphatase in COS‐Ce/600 were significantly higher than that in COS‐Ce/0 (P < 0.05) respectively. On the contrary, cumulative mortality was the lowest in COS‐Ce/600 following 14 days exposure to Vibrio splendidus (P < 0.05). In conclusion, these results confirmed that dietary COS‐Ce had beneficial effects on growth performance, nonspecific immunity and disease resistance of sea cucumber.