紫球藻胞外多糖抗氧化和免疫调节活性的研究

采用密闭微波法降解紫球藻胞外多糖。通过测定多糖对羟自由基OH·、超氧阴离子O2-·和二苯代苦味酰基自由基DPPH·的清除效果及对小鼠单核-巨噬细胞RAW264.7和淋巴细胞的增殖效果,考察低分子量紫球藻多糖的体外抗氧化活性和免疫调节活性。结果表明,在一定的浓度范围内,降解后的紫球藻多糖对OH·、O2-·和DPPH·的清除能力与浓度呈正相关性,它们的半抑制率IC50分别为0.619,0.114和0.015mg/mL,表明紫球藻胞外多糖具有体外抗氧化活性。溶液浓度在50～200μg/mL的多糖都表现出体外免疫促进作用,与对照组相比,100μg/mL的多糖对巨噬细胞RAW264.7的促进作用最大,其增值指数为1.72。200μg/mL的多糖对脾淋巴细胞的增值指数为2.11。     

温度对小黄鱼体内抗氧化酶及消化酶活性的影响

为了深入了解不同温度条件下小黄鱼的应激反应,为小黄鱼的人工养殖提供参考。本次研究以人工繁殖的4.5月龄小黄鱼为研究对象,采用不同水温（27℃、29℃、31℃、32℃、33℃）进行养殖,以此确定小黄鱼的高温致死温度,并检测不同养殖水温条件下小黄鱼肝脏和肠道中抗氧化酶（超氧化物歧化酶SOD、过氧化氢酶CAT）和消化酶（脂肪酶LPS、淀粉酶AMS）的活性,比较小黄鱼不同组织中4种酶活性的变化情况。结果显示,32℃的养殖水温为小黄鱼的初始致死温度;养殖温度对小黄鱼肝脏和肠道中几种酶的活性均存在显著影响（P<0.05）:抗氧化酶和消化酶活性均随着养殖水温的升高呈现先升高后下降的趋势（除了肠道中的CAT）。另外,肝脏中抗氧化酶活性强于消化酶,而肠道中则相反,表明肝脏侧重于机体免疫反应,而肠道更侧重于食物消化。通过二次项回归方程拟合了温度与酶活性的回归方程,只有肝脏和肠道中的LPS活性与温度的回归方程有意义,决定系数r2分别为0.89和0.93（在0.85以上）。此外,肝脏中LPS活性与实验鱼体质量呈极显著正相关关系（P<0.01）,研究结果为小黄鱼专用配合饲料的研发提供了重要参考。研究还得出,养殖水温29℃和31℃时,进行小黄鱼养殖可取得较好效果,研究结果可为小黄鱼快速养成提供指导。     

Preparation by enzymolysis and bioactivity of iron complex of fish protein hydrolysate （Fe-FPH）from low value fish

Preparation of Fe^2＋ chelate of fish protein hydrolysate （Fe-FPH） obtained from low value fish proteins was introduced and its bioactivity was studied by compound enzymolysis. The optimum conditions for hydrolysate chelating Fe^2＋ are DH （degree of hydrolysis） at 5%, pH 7.0, 20℃ and 15 min chelating time for FM （material not being defatted）. Four types of Fe-FPH including CA （deposit after chelating）, CB （deposit in 50% of absolute ethanol solution）, CC （suspended deposit in 80% of absolute ethanol solution）, and CD （bottom deposit in 80% of absolute ethanol solution） were fractionated with absolute ethanol from FM. Structural analysis through infra-red spectrum revealed that Fe^2＋ was combined strongly with amino-group and carboxyl-group in each chelate and each Fe^2＋ could form two five-member ring structures. All of the four chelates were shown more significant antioxidative activity and can be used as natural hydrophobic and hydrophilic antioxidant. Among all the chelates, the CB possesses the most effective antioxidative activity at 92% as high as that of a-tocopherol. Among all Fe-FPHs, only CD showed the most effective antibacterial activity against Escherichia coli, Staphylococcus aureus, Salmonella typhi, and Bacillus subtilis and can be used as natural antibacterial. It provides a more effective way for utilization of low value fish proteins and key information of Fe-FPH as additive in food industry.     

Effect of sodium chloride and cadmium on the growth,oxidative stress and antioxidant enzyme activities of Zygosaccharomyces rouxii

Zygosaccharomyces rouxii is a salt-tolerant yeast species capable of removing cadmium(Cd) pollutant from aqueous solution. Presently, the physiological characteristics of Z. rouxii under the stress of sodium chloride(NaCl) and Cd are poorly understood. This study investigated the effects of NaCl and Cd on the growth, oxidative stress and antioxidant enzyme activities of Z. rouxii after stress treatment for 24 h. Results showed that NaCl or Cd alone negatively affected the growth of Z. rouxii, but the growth-inhibiting effect of Cd on Z. rouxii was reduced in the presence of NaCl. Flow cytometry assay showed that under Cd stress, NaCl significantly reduced the production of reactive oxygen species(ROS) and cell death of Z. rouxii compared with those in the absence of NaCl. The activities of superoxide dismutase(SOD), catalase(CAT), and peroxidase(POD) of Z. rouxii were significantly enhanced by 2%–6% NaCl, which likely contributed to the high salt tolerance of Z. rouxii. The POD activity was inhibited by 20 mg L-1Cd while the SOD and CAT activities were enhanced by 8 mg L-1 Cd and inhibited by 20 mg L-1 or 50 mg L-1 Cd. The inhibitory effect of high-level Cd on the antioxidant enzyme activities of Z. rouxii was counteracted by the combined use of NaCl, especially at 6%. This probably accounted for the decrease in Cd-induced ROS production and cell death of Z. rouxii after incubation with NaCl and Cd. Our work provided physiological clues as to the use of Z. rouxii as a biosorbent for Cd removal from seawater and liquid highly salty food.     

外源一氧化氮和金属铜对海洋微藻抗氧化系统的影响

以亚心形扁藻为研究对象,通过添加不同浓度一氧化氮(NO)(1.4×10-10～1.4×10-5 mol/L)培养,探讨NO对微藻生长、H2O2和MDA含量以及SOD活性的影响;通过对只添加金属Cu、同时添加Cu与NO实验,对微藻的H2O2和MDA含量以及SOD活性的影响进行了对比研究。结果表明,藻细胞活性氧的代谢有随NO浓度增加的趋势,同时细胞内的抗氧化酶SOD活性和MDA含量都有相应的变化,NO作为环境的"刺激物"提高了细胞内的活性氧水平,对抗氧化系统起到激活或伤害作用。在Cu胁迫下,藻细胞活性氧代谢明显提高,SOD活性提高,并导致细胞膜脂过氧化;但外源NO能明显降低活性氧的水平,并恢复SOD活性和MDA含量,对抗氧化系统起到保护作用,提高微藻的抗氧化能力。研究表明,NO在海洋浮游植物中扮演着"双重角色"——氧化剂和抗氧化剂,从而对微藻的生长起到重要的调节作用。     

Treating ballast water with hydroxyl radical on introduced organisms

1 INTRODUCTION The introduction of invasive marine species into new environments by ship’s ballast water, by attaching to ships’ hulls or via other vectors has been recognized as one of the four major threats to the world’s oceans by Global Environment…     

化学诱导对2株海洋真菌次级代谢产物及其生物活性的影响

【目的】以2株海洋真菌为研究对象,探究不同种类及浓度的化学表观遗传修饰剂对其次级代谢产物化学多样性和生物活性的影响。【方法】基于化学表观遗传修饰策略,采用两种不同的化学表观遗传修饰剂5-氮杂胞嘧啶核苷(5-azaC)和丁酸钠,分别对Penicillium sp.019和Aspergillus terreus ZN4-5-4两株海洋来源真菌进行表观遗传修饰。通过观察真菌在培养基中的生长状态变化,并根据薄层层析(TLC)指纹图谱、高效液相色谱(HPLC)图谱、乙酰胆碱酯酶(AChE)抑制、1,1-二苯基-2-三硝基苯肼(1,1-Diphenyl-2-picryl-hydrazyl,DPPH)自由基清除和卤虫致死活性筛选模型等技术手段,从中筛选出代谢产物丰富、AChE抑制活性与抗氧化活性产物丰富且毒性小的发酵培养条件。【结果】添加1 mmol/mL的丁酸钠诱导剂可使菌株019和ZN4-5-4的次生代谢产物的种类增加,且同时能提高该菌株次级代谢产物的AChE的抑制活性和DPPH自由基清除活性。因此,确定菌株019和ZN4-5-4的最优发酵条件均是添加浓度为1 mmol/mL丁酸钠的PDB培养基。【结论】化学表观遗传修饰策略指导下,添加诱导剂5-azaC或者丁酸钠对2株海洋真菌次生代谢产物的产生有影响。     

植物乳杆菌(Lactobacillus plantarum)对云龙石斑鱼生长、抗氧化性能及血清生化水平的影响

植物乳杆菌(Lactobacillus plantarum)常作为饲料添加剂,可抑制鱼类病原菌生长,增加机体对饲料的消化和吸收。探讨了饲喂不同添加量的植物乳杆菌对云龙石斑鱼(Epinephelus moara♀×E.lanceolatus♂)生长性能、肠道消化酶活性、肠道和肝脏抗氧化能力、血清生化水平的影响,以期为植物乳杆菌在其健康养殖中的应用提供依据。结果显示,随植物乳杆菌的饲喂量增加,云龙石斑鱼的终末均重(FAW)、增重率(WGR)、特定生长率(SGR)和肠道中淀粉酶(AMS)、脂肪酶(LPS)和糜蛋白酶(CHY)、总抗氧化能力(T-AOC)、过氧化氢酶(CAT)、还原型谷胱甘肽(GSH)、超氧化物歧化酶(SOD)的变化均呈现先升高后降低的趋势。而随着植物乳杆菌的饲喂量增加,云龙石斑鱼的血清中总蛋白(TP)、白蛋白(Alb)、胆固醇(TC)、碱性磷酸酶(AKP)、酸性磷酸酶(ACP)、超氧化物歧化酶(SOD)、丙二醛(MDA)均略降低,甘油三酯(TG)略升高,但与对照组差异不明显(P>0.05)。当添加量为1×10⁵CFU/g时,云龙石斑鱼的各生长指标和抗氧化性能均优于对照组,其中增重率显著提高了19.8%(P<0.05),肠道T-AOC、SOD分别显著升高了93.2%、2.9%(P<0.05),肝脏MDA显著降低了32.7%(P<0.05)。当添加量为1×10⁷CFU/g时,云龙石斑鱼的终末均重、增重率、特定生长率均显著高于对照组和其余添加组(P<0.05);饲料系数显著低于对照组(P<0.05);肠道的AMS、LPS、CHY活性分别显著提高了13.0%、166.9%、61.1%(P<0.05);但肠道和肝脏的抗氧化性能变化不明显,仅肝脏MDA显著降低了23.9%(P<0.05)。由此表明,饲料中添加一定量的植物乳杆菌可以提高肠道消化能力和抗氧化能力,促进云龙石斑鱼的生长,但对其血清生化水平无显著性影响,不会对鱼体代谢产生不良作用。     

金鼓鱼幼鱼对蛋白质的适宜需要量研究

为确定金鼓鱼(Scatophagus argus)日粮中蛋白质的适宜添加量,作者以酪蛋白为蛋白源、鱼油为脂肪源,配制脂肪水平为8%,蛋白水平分别为24%、28%、32%、36%、40%的5种配合饲料(D1~D5),在海上网箱中以初始体质量为4.3 g左右的幼鱼开展养殖实验。8周后,测定实验鱼增质量率、特定生长率、蛋白质效率、饲料系数、消化酶和一些免疫及抗氧化指标。结果显示,随着饲料蛋白水平的升高,金鼓鱼的增质量率、特定生长率、蛋白质效率等均呈现先上升后下降的趋势,以D3组最高,3项指标均显著高于D1、D2组;饵料系数呈现先下降再上升趋势,以D3组最低,且显著低于D1、D2组。饲料蛋白水平也显著影响金鼓鱼的相关生理生化指标,其中D3和D4组的全鱼蛋白含量、消化道蛋白酶活性和一些免疫抗氧化指标最高,全鱼脂肪含量最低。根据饲料蛋白含量与增质量率的线性回归分析,确定金鼓鱼幼鱼饲料中的适宜蛋白水平为35%。