酵母免疫多糖对受免斑点叉尾免疫应答的增强作用

在饲料中添加定量的酵母免疫多糖,投喂注射接种了福尔马林灭活的嗜水气单胞菌(Aeromonas hydrophila)菌苗的斑点叉尾(Ictalurus punctatus),28 d后通过测定供试鱼的增重量、血清中凝集抗体效价、溶菌酶活性、谷丙转氨酶活性(SGPT)、血清总蛋白含量、白细胞吞噬活性以及活菌攻毒后的免疫保护率(RPS),探讨了酵母免疫多糖对受免斑点叉尾免疫应答的增强作用。结果表明,用添加200.0 mg/(kg.d)酵母免疫多糖的饲料投喂受免斑点叉尾,不仅可以使受免斑点叉尾对灭活嗜水气单胞菌的免疫应答水平提高,增强抵抗嗜水气单胞菌人工感染的能力,而且还具有一定的促进生长和改善肝功能的作用。     

Protection of Procambarus clarkii against white spot syndrome virus using recombinant subunit injection vaccine expressed in Pichia pastoris

ABSTRACT:   The potentiality of injection vaccine against white spot syndrome virus (WSSV) in crayfish Procambarus clarkii was investigated. WSSV envelope proteins VP19 and VP28 were expressed in yeast Pichia pastoris GS115. The purified recombinant proteins (2 µg/g of crayfish) were injected intramuscularly, and the same dose injected as a booster shot on fifth day after vaccination. The vaccinated crayfish were divided into two even groups and later challenged orally by WSSV-infected dead crayfish muscle (2 g/individual) on the third and 21st days after the booster shot. The relative percent survival (RPS) in the third-day group was the highest in VP28 (91%), followed by VP19 + VP28 (84%), and VP19 (45%). The RPS for the 21st-day group was the highest in VP28 (78%), followed by VP19 + VP28 (76%), and VP19 (17%). Development of vaccine by using recombinant proteins VP19 and VP28 expressed in yeast is feasible.     

Use of marine yeasts as an available diet for mass cultures of Moina macrocopa

A 4‐week feeding trial was conducted to test the use of marine yeasts purified from seawater and sediments as a dietary source in cultivating a cladoceran, Moina macrocopa, a potential alternative live food for fish larvae. Optimal growth conditions of two yeast strains were obtained for NaCl concentration, pH and temperature. Moina macrocopa was cultivated using different diets: marine yeasts (Debaryomyces hansenii Yeast‐14 and Candida austromarina Yeast‐16) and a commercial diet (Erythrobacter sp. Sπ‐I). The essential amino acids of both the yeast strains were nearly as great as those in M. microcopa. Further, the yeast‐fed M. macrocopa had essential amino acid profiles similar to the documented values for rotifers and Artemia enriched in microalgae and commercial diets. Erythrobacter sp. Sπ‐I lacked n‐3 polyunsaturated fatty acids, 20:5n‐3 and 22:6n‐3, which were also low but detected in both yeasts. An increase in the 20:5n‐3 [eicosapentaenoic acid (EPA)] and 22:6n‐3 [docosahexaenoic acid (DHA)] levels, compared with the levels in yeast strains themselves, was more pronounced in the 22:6n‐3 level of Moina fed the C. austromarina, resulting in a high DHA:EPA ratio. When the Moina diets were switched, their δ¹³C values shifted gradually towards the values of the switched diets. Diet switch from Erythrobacter sp. Sπ‐I to C. austromarina Yeast‐16 resulted in a more rapid turnover of Moina tissue carbon than that in the inverse case. When fed a mixed diet, the δ¹³C values of Moina tissue approached the value of marine yeasts immediately. These temporal changes in the δ¹³C values of Moina tissue indicate the preferential ingestion of marine yeasts and a selective assimilation of the carbon originated from marine yeasts. These findings suggest that marine yeasts, particularly C. austromarina Yeast‐16, are highly available to mass cultures of M. macrocopa, providing better nutritional and dietary values than the commercial diet (Erythrobacter sp. Sπ‐I).     

接种密度和面包酵母投喂量对蒙古裸腹溞种群变动和生产量的影响

在室内水槽中进行蒙古裸腹(Moinamongolica)的大量培养,研究不同接种密度和面包酵母投喂水平对该种群变动和生产量的影响。所用蒙古裸腹为20世纪90年代采自山西省晋南半咸水湖、在海水中长期培养保存下的种。实验1中,接种密度分别设为338ind/L、145ind/L和88ind/L3个水平,面包酵母投喂量为5mg/L,每天4次,各设3个重复。实验2中,面包酵母投喂量分别为30mg/(L·d)、20mg/(L·d)和10mg/(L·d)3个水平,蒙古裸腹接种密度为900ind/L,各设3个重复。实验结果表明:(1)在一定范围内蒙古裸腹培养产量随接种密度增加而增加,当接种密度低于88ind/L时,接种的蒙古裸腹较难形成稳定的种群。(2)当使用面包酵母(Bakeryeast)作为培养蒙古裸腹的惟一食物时,食物需要量随种群密度的变化而变化。当密度低于1500ind/L时,适宜的面包酵母投喂水平为20mg/(L·d);当密度超过5000ind/L时为30mg/(L·d)。上述结果显示,接种密度和面包酵母投喂量对蒙古裸腹种群变动和生产量具有重要影响。建议在蒙古裸腹培养生产中接种密度高于150ind/L,面包酵母投喂量为0 8×10-5～1 3×10-5g/(ind·d)。     

人C型溶菌酶发酵与分离纯化工艺的研究

[目的]研究毕赤酵母菌株(Pichia GS115)表达人C型溶菌酶(H-LYZ)的发酵与分离纯化的工艺.[方法]探讨酵母生长规律,研究不同菌体浓度、不同诱导时间对蛋白产量的影响,并且探讨不同pH条件对DEAE-SepharseFF和CM-Sepharose FF层析分离纯化目标蛋白的影响.[结果]毕赤酵母菌株表达H-LYZ的最佳发酵条件为:发酵24h用浓度1％甲醇诱导,以后每12 h用浓度1％甲醇诱导,共6次,在此条件下可获得高产量目标蛋白.DEAE-Sepharose FF和CM-Sepharose FF分离纯化目标蛋白的最佳pH分别为6.65和6.50.[结论]该研究可为人C型溶菌酶的工艺化生产提供理论指导.     

微生物学大实验模块“天然酵母菌的筛选、扩大培养和鉴定”的教改研究与实践

以“天然酵母菌的筛选、培养和选育”为主线,设计了一个由4个相互关联、前后衔接一体的实验组成的综合性研究型实验模块,进行教学实践与改革。实验模块主要包括：培养基的配制、分装与灭菌,酵母菌的分离与纯化,酵母的扩大培养,菌种的鉴定。强化实践,提高学生对于知识的综合应用能力。     

混菌固态发酵降解羽毛生产饲料蛋白质工艺研究

试验主要以酵母菌和淡紫拟青霉为发酵微生物,羽毛粉和麸皮为发酵底物,采用混菌固态发酵方式,探究基质初始水分以及麸皮与羽毛粉的比例等因素对饲料蛋白质生产的影响。试验结果表明,麸皮:水=1:1时最适合酵母菌的生长;麸皮:水=1:0.8时,最适合淡紫拟青霉的生长。以1:1的水料比,当麸皮:羽毛粉=7:3时,最适合酵母菌的生长;而当麸皮:羽毛粉=6:4时,最适合淡紫拟青霉的生长。因此,混菌固体发酵降解羽毛生产饲料蛋白质最佳初始水分是1:1,麸皮与羽毛比为7:3。     

酵母硒对凡纳滨对虾生长和抗氧化性能的影响

【目的】研究饲料中添加不同水平酵母硒对凡纳滨对虾生长和抗氧化性能的影响.【方法】选取初始体质量为(0.41±0.01)g的凡纳滨对虾Litopenaeus vannamei 720尾,随机分为6个试验组,每组3个重复,每个重复40尾虾,在室内循环水养殖系统(容积250 L)中进行为期56 d的生长试验.在基础饲料中分别添加硒(酵母硒)0、0.15、0.30、0.45、0.60、1.00 mg·kg-1,制成6种不同硒水平的半纯化饲料,实测饲料硒质量分数分别为0.25、0.41、0.56、0.68、0.84、1.20 mg·kg-1.【结果和结论】结果表明,添加酵母硒对凡纳滨对虾增质量率、特定生长率、饲料系数、成活率均无显著影响(P0.05),随着饲料硒水平的增加,凡纳滨对虾的增质量率和特定生长率不断升高,在0.84 mg·kg-1硒组达到最大值.各试验组对虾肝胰腺超氧化物歧化酶和总抗氧化能力均高于对照组,而丙二醛含量均低于对照组,但差异不显著(P0.05).以增质量率和特定生长率为评价指标,通过二次线性回归分析表明,在该试验条件下凡纳滨对虾饲料中硒含量为0.98 mg·kg-1时其生长性能最高.     

不同型号酵母浸粉及补糖对异养微藻生长的影响

微藻在FM802、FM902和FM760 3种不同型号的安琪酵母浸粉配制的培养基中进行异养生长,培养条件为28℃、200 r/min的摇床中进行暗培养,培养过程中补糖1次,每隔24 h取样20 mL进行分析检测。结果表明:补糖情况下,型号为FM802的酵母浸粉比FM902培养微藻的效果稍好,FM760培养效果稍差;补糖有利于微藻干物质重的增加;在pH值为4.2~5.5的培养基中,微藻仍能快速生长。