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。     

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。     

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.     

Effects of different molecular weights of chitosan on methane production and bacterial community structure in vitro

As a new feed additive, chitosan has been shown in recent years to have a certain role in reducing methane emissions from the gastrointestinal tracts of ruminants. However, the effects of chitosan with different molecular weights on rumen fermentation, methane production and bacterial community structure are not yet clear. A basal diet without chitosan served as the control(CTL), and the treatment diets were supplemented with chitosan with different molecular weights: 1 000(1K), 3 000(3K), 5 000(5K), 50 000(5W) and 200 000(20W) dry matter(DM). Six fermentation units per treatment were established. Gas chromatography was used to measure the concentrations of methane, H_2 and volatile fatty acids(VFAs). The bacterial 16S rRNA genes were sequenced with an Illumina MiSeq platform and analysed to reveal the relative abundances of bacterial community taxa. The results showed that the propionate proportion was significantly increased by the addition of chitosan with different molecular weights(P0.05), while methane production and the acetate proportion were significantly decreased(P0.05). The relative abundances of Rikenellaceae_RC9_gut_group and Prevotellaceae_UCG_003 were significantly increased in the 3K chitosan group compared with the CTL group, whereas the relative abundance of Ruminococcaceae_NK4A214_group was significantly decreased(P0.05). Correlation analyses of the relative abundances of the bacterial genera showed that Prevotella was positively related to propionate production(P0.05). In conclusion, 3K chitosan could reduce methane production by replacing fibrolytic bacteria(Firmicutes and Fibrobacteres) with amylolytic bacteria(Bacteroidetes and Proteobacteria) in the bacterial community structure.     

膨润土改性及对水中Cr(Ⅵ)吸附性能的研究

目的 提高膨润土对水中Cr（Ⅵ）的吸附性能。方法 采用氢氧化钠和壳聚糖对膨润土进行改性，分别得到碱改性膨润土（B-NaOH）、壳化膨润土（B-CS）和壳化碱改性膨润土（B-NaOH-CS）。以钠基膨润土（B）为对照，利用红外光谱仪、扫描电镜和比表面积分析仪表征3种改性膨润土的理化性质，研究其对Cr（Ⅵ）的吸附性能。结果 B-NaOH-CS中出现了强N—H吸收峰以及增强的C—H对称弯曲峰，同时B-NaOH-CS表面片状结构卷曲分散，层间孔隙增多，比表面积是其他膨润土的1.2倍以上。当Cr（Ⅵ）质量浓度为50 mg·L^-1时，B-NaOH-CS对Cr（Ⅵ）的平衡吸附量为1.03 mg·g^-1，分别是B-CS、B-NaOH的1.26、1.84倍。描述膨润土吸附Cr（Ⅵ）的动力学过程，准二级动力学模型优于准一级动力学模型；描述膨润土吸附Cr（Ⅵ）的热力学过程，Langmuir等温模型优于Freundlich等温模型。热力学参数△H>0、△G<0、△S>0，表明膨润土吸附Cr（Ⅵ）为吸热、自发、无序反应。B-NaOH在pH=7.0时对Cr（Ⅵ）的吸附量最大，B-CS、B-NaOH-CS在pH = 3.0时对Cr（Ⅵ）的吸附量最大。结论 B-NaOH-CS对Cr（Ⅵ）的吸附效果最好，改性膨润土对去除Cr（Ⅵ）污染有重要作用。     

壳聚糖硒对蛋雏鸡生长、组织硒含量及血清肝酶活性的影响

为探索壳聚糖硒对蛋雏鸡生长、组织硒含量及血清肝酶活性的影响,选取135只1日龄SPF蛋雏鸡,随机分为3组:一组为对照组,饲喂基础日粮;二组和三组在基础日粮中分别添加硒水平为0.15 mg/kg的亚硒酸钠和壳聚糖硒。结果显示:14日龄时亚硒酸钠组和壳聚糖硒组的耗料量、血清碱性磷酸酶(AKP)活性和胸肌、心肌、肾脏、全血的硒含量显著高于对照组(P<0.05),壳聚糖硒组心肌和肝脏的硒含量显著高于亚硒酸钠组(P<0.05);28日龄时壳聚糖硒组与亚硒酸钠组的体重、血清AKP活性和肝脏、肾脏、胸肌、心肌、全血的硒含量显著高于对照组(P<0.05),壳聚糖硒组的耗料量和肾脏、心肌、全血的硒含量显著高于亚硒酸钠组(P<0.05)。研究表明:壳聚糖硒能促进鸡的生长和硒的沉积,提高血清AKP活性,且效果优于亚硒酸钠。     

链霉菌和壳聚糖对淮山土壤微生物活性与群落的影响

为探究链霉菌30702和壳聚糖对淮山土壤微生物活性与群落的影响,采用比色法和滴定法观测土壤酶活性的变化,采用高通量测序技术和Trimmomatic等软件,分析土壤微生物群落中细菌和真菌的发展变化。结果表明,链霉菌、壳聚糖、土壤原始微生物和培养时间等4个因素对淮山土壤的脲酶和过氧化氢酶活性均有显著影响,壳聚糖浓度在0~10.0 g/kg内,土壤酶活性随壳聚糖浓度的增大而提高;随时间的延长,先升再降;壳聚糖与链霉菌之间存在交互效应。链霉菌和壳聚糖能增加淮山土壤细菌的物种丰度,减少细菌的多样性,增加艾德昂菌属（Ideonella）和纤维弧菌（Cellvibrio）等有益菌属的相对丰度,而对真菌的物种丰度和多样性的影响较小,与对照之间的差异不显著,土壤优势菌属为腐质霉属（Humicola）、枝孢属（Cladosporium）、鞘氨醇单胞菌属（Sphingomonas）和uncultured_bacterium_ c_Subgroup_6。其中,链霉菌与2.5 g/kg壳聚糖复合处理的细菌物种丰度最大,细菌多样性最少。