长效益生素菌株的筛选及生产工艺的初步研究

益生素是一种能够替代抗生素的绿色饲料添加剂。报道了通过对益生素菌种的初筛和复筛、菌株药敏实验、菌体吸附方法及产品的中试研究 ,确定了生产工艺 :采用筛选的地衣芽孢杆菌YS 0 2 ,在特定的培养基和培养条件下 18～ 2 2h ,通过 10 %CaCO3 吸附、过滤、干燥获得微生态制剂 ,其活菌数≥ 15 0亿 / g。实验证明 ,YS 0 2活菌制剂可替代抗生素防治动物习惯性腹泻。     

医学真菌菌株保藏方法概述

近年来,随着真菌感染的患者急剧增加,真菌病已经成为一个重要的公共卫生问题。对医学真菌流行病学、致病机理、耐药机理以及防治策略的研究尤为重要,而所有研究的基础都建立在对医学真菌的进行有效的保藏。医学真菌的保藏方法很多,包括传代法、蒸馏水法、冷冻干燥保藏法等,保藏方法的选择因实验室条件、菌种和研究要求不同而不同。本文对目前常用的几种医学真菌菌种保藏方法的优缺点及其应用等做了综述。     

苏云金芽孢杆菌高效价杀虫剂的研究进展

苏云金芽孢杆菌制剂是目前应用广泛而有效的一种微生物杀虫剂.本文从菌种分离筛选、构建遗传工程菌、突变筛选及定向进化等方面介绍了获得苏云芽孢金杆菌高效价杀虫的几种研究策略,并对各种策略的进展及应用前景进行了简要综述.     

现代生物技术与饲用微生态制剂

近年来,使用抗生素的副作用越来越多地受到关注,世界上许多国家已出台相应政策来控制抗生素的使用。但是由于养殖行业的迅猛发展,养殖密度加大,养殖动物病害发病的风险提高,急需可替代抗生素的新型绿色饲料添加剂产品。微生态制剂作为一种新型绿色饲料添加剂,在养殖行业发挥了重要作用。随着饲用微生态制剂产品研发的深入,现代生物技术在提升微生态制剂的理论和应用研究方面发挥着重要作用。PCR、核酸分子杂交、基因工程以及组学等技术已应用于微生物菌种鉴定、基因改良、作用机理等研究中。本文对饲用微生态制剂的研发现状进行了阐述,并综述了现代生物技术在饲用微生态制剂研究中的应用。     

皮肤益生素霜与粉刺消护肤霜在治疗痤疮中的比较

观察皮肤益生素霜治疗痤疮的疗效。选择I- IV 度痤疮患者各40 例,共计160 人分二组试用,每组80人。结果使用皮肤益生素霜的患者总有数率为76.2% ,使用粉刺清护肤霜的患者的总有效率为62.5% ,前者与后者相比疗效提高13.3% 。皮肤益生素霜是微生态制剂,以菌治菌,达到消炎的目的。其内含有多种营养成分,作用柔和,宜可长期使用,即治病又可美容     

微生态制剂的临床研究现状

在简述微生态制剂的类型和菌种的基础上,从屏障作用、营养作用、免疫调节作用、抗肿瘤作用等方面综述了微生态制剂的作用机制和微生态理论,并从预防治疗各种腹泻、结肠癌、动力障碍性疾病、肝硬化以及某些自身免疫性疾病等方面综述了微生态制剂在临床上的应用,继而指出了应用注意事项,展望了前景。     

生态工程领域微生物菌剂研究进展

阐述了微生物菌剂在生态工程领域的应用范围、效果和国内外的研究状况,总结了目前微生物菌剂研制中菌种的选育方法和常用的几类菌,并通过系统举例介绍了乳酸菌、酵母菌、光合细菌、芽孢杆菌这些常用菌种在污染物处理方面的效果及在废水处理生态工程方面的相关研究进展。同时,阐明了微生物混合培养技术在复合菌剂研究中的重要性,对微生物菌剂的作用机理进行了探讨,指出了目前关于微生物菌剂的研究大多只是集中于应用效果方面,而对作用机理研究得不够深入,以及复合菌系中微生物相互作用和影响的复杂性,并对此提出了一些建议。最后,对微生物菌剂的应用前景进行了展望。     

枯草芽孢杆菌微生态制剂发酵研究进展

微生态制剂是饲用抗生素的绿色有效替代品。枯草芽孢杆菌在逆境中可形成抗逆性强的芽孢,在生产和应用过程中保持高活性,是一种高效的微生态制剂菌种。提高枯草芽孢杆菌活菌数及芽孢率是保证微生态制剂产品质量的关键。本文综述了枯草芽孢杆菌芽孢形成的分子生物学机制及影响芽孢形成的重要因素,进一步比较枯草芽孢杆菌微生态制剂不同发酵方式的特点,重点阐述了提高枯草芽孢杆菌有效生物量的工艺优化,最后介绍了枯草芽孢杆菌微生态制剂的应用,并对将来研究思路进行了讨论。     

功能性便秘的微生态学的防治策略

功能性便秘是儿童时期常见的消化系统功能性疾病,其主要特点为经常排便困难和疼痛,可伴有大便失禁和(或)腹痛。其不仅影响患儿的生活质量,还给患儿带来严重的心理及精神负担。其具体发病机制尚不完全清楚,临床常采用综合治疗方法,但一部分患儿治疗效果欠佳。近年来随着对肠道微生态学的研究逐渐增多,了解到肠道菌群失调与便秘的发生有重要关联,人们也在开始尝试将益生菌等微生态制剂应用于功能性便秘患儿,但其治疗效果有待进一步证实。本文主要通过收集有关功能性便秘患儿应用益生菌及益生元等制剂后的效果,分析益生菌等微生态制剂在治疗功能性便秘患儿的效果,以指导未来临床对功能性便秘患儿的治疗。     

苏云金芽孢杆菌的紫外线诱变及筛选实验初步设计

苏云金芽孢杆菌(Bacillus thuringiensis)由于其自身特点,由其制成的杀虫剂已成为目前微生物防治害虫的重要手段之一。虽然苏云金芽孢杆菌制剂目前应用效果良好,但也存在一些隐患:长期使用该制剂会使害虫抗性增加,因此寻找新的高毒力菌株势在必行。本文以苏云金芽孢杆菌的库尔斯塔克亚种为研究对象,设计实验以紫外线诱变的方法获得毒力更强的菌种。     

Film coating of seeds with Bacillus cereus RS87 spores for early plant growth enhancement

The plant growth-promoting rhizobacterium Bacillus cereus RS87 was previously reported to promote plant growth in various crops in both greenhouse and field trials. To apply as a plant growth promoting agent with practical use, it is essential to ease the burden of routine preparation of a fresh suspension of strain RS87 in laboratory. The objectives of this study were to investigate the feasibility of film-coating seeds with B. cereus RS87 spores for early plant growth enhancement and to reveal the indoleacetic acid (IAA) production released from strain RS87. The experiment consisted of the following 5 treatments: nontreated seeds, water-soaked seeds, film-coated seeds, seeds soaked with vegetative cells of strain RS87, and film-coated seeds with strain RS87 spores. Three experiments were conducted separately to assess seed emergence, root length, and plant height. Results showed that both vegetative cells and spores of strain RS87 significantly promoted (P < or = 0.05) seed emergence, root length and plant height over the control treatments. The strain RS87 also produced IAA. In conclusion, the film coating of seeds with spores of B. cereus RS87 demonstrated early plant growth enhancement as well as seeds using their vegetative cells. IAA released from strain RS87 would be one of the mechanisms for plant growth enhancement.     

糖化酶生产菌株种子制备工艺优化

目的：缩短糖化酶工业生产菌株黑曲霉A.nigerCICIM GB0506的种子制备周期。方法：对该菌的活化培养基配方及种子制备方式进行改良,并通过L9（3^4）正交试验对其液体培养方法进行优化。结果：在固体活化培养基中添加0.2%的酵母粉及1%的玉米淀粉,有利于加速菌体生长;加入40粒,粒径3-4mm的玻璃珠130r/min,摇床培养可以获得更为分散、均一的种子液;液体种子制备的较优条件为初始pH4.5,装液量90mL,琼脂加量0.1%,培养天数5d。结论：新的制备工艺使糖化酶种子制备时间较现在工业生产上使用的工艺缩短了4d,进行后续糖化酶摇瓶发酵产酶水平是原工艺的1.18倍。     

Yellow fever vaccine: Comparison of the neurovirulence of new 17D-204 Stamaril™ seed lots and RK 168-73 strain

The neurovirulence of two new candidate 17D-204 Stamaril? working seed lots and that of two reference preparations were compared. The Stamaril? working seed lots have been used for more than twenty years for the manufacturing of vaccines of acceptable safety and efficacy. The preparation designated RK 168-73 and provided by the Robert Koch Institute was used as a reference. It was confirmed that RK 168-73 strain was not a good virus control in our study because it has a very low neurovirulence regarding both the clinical and histopathological scores in comparison with Stamaril? strain and is not representative of a vaccine known to be satisfactory in use. The results were reinforced by the phenotypic characterization by plaque assay demonstrating that RK 168-73 was very different from the Stamaril? vaccine, and by sequencing results showing 4 mutations between Stamaril? and RK 168-73 viruses leading to amino acid differences in the NS4B and envelop proteins.     

Quantification of two variant strains contained in freeze-dried Japanese BCG vaccine preparation by real-time PCR.

Japanese bacillus Calmette-Guerin (BCG) vaccine preparation contains two types of variant strains, Type I, which has a 22-base-pair deletion in the RD16 region, and Type II, which has an identical sequence to those of other BCG strains. In this study, we established a method to quantify the percentage of variant strain Type II contained in freeze-dried BCG product with real-time PCR. With this method we examined the master seed lot Tokyo 172, two secondary seed lots, Tokyo 172-1 and Tokyo 172-2, which were produced from Tokyo 172, and four commercial lots produced form Tokyo 172-1. Tokyo 172, Tokyo 172-1, and Tokyo 172-2 contained 55.1%, 19.5%, and 3.6% of Type II variant strain, respectively. Commercial lots contained 1.5%, 4.5%, 7.4%, and 4.3% of Type II variant strain, respectively. These results indicated that the two variant strains contained in the master seed lot continued to coexist in subsequently produced lots with a decrease in population of variant strain Type II. This method would be useful for quality control of commercial Japanese BCG vaccine preparations.     

外源DNA直接导入小麦及其在育种上的应用

本研究选用两个白粒小麦品种作供体,提取其总ＤＮＡ,采用花粉管直接携带法导入７５（１９８）红粒品种受体植株。ＤＮＡ导入的第一代（Ｄ１）,目的性状的转化频率为１．７５％和２．９４％。Ｄ２代变异率显著低于Ｄ１代。对Ｄ１,Ｄ２代所得目的性状变异后代,按照常规育种程序进行Ｄ３代观察与鉴定,得到已稳定遗传的后代,从中选取保持原品种其它优良性状而籽粒为的白色的变异类型混合脱粒,获得７５（１９８）改良新品系。     

腺苷高产菌株Bacillus subtilis DI4-24退化的遗传机制与控制研究

经人工诱变获得的腺苷高产菌株Bacillus subtilis DI4-24很容易发生退化,通过分析回变菌株的遗传性状和考察理化条件对回复突变的影响,研究退化的机制,找到控制的方法。在选择培养基上筛选回变菌株,生长谱法分析其遗传型;于743 nm条件下分光光度法测定回复突变细胞内积累的特有红色物质,研究各理化条件对回复突变的影响。退化原因是由于组氨酸缺陷型发生回复突变所致,突变菌株腺苷产量大幅度下降;降低保藏温度、减少传代次数可以有效地减缓菌种的衰退;种子制备过程中适当降低培养温度2℃~3℃、缩短培养时间(8 h)、降低培养基pH至中性偏酸性(6.5)以及加入胡萝卜素、叶酸等物质都可以有效降低回变率,在此条件下制备的种子生产性能得到提高,其腺苷产量可比对照提高9%。     

Recalcitrancy and a new kind of epicotyl dormancy in seeds of the understory tropical rainforest tree Humboldtia laurifolia (Fabaceae, Ceasalpinioideae)

We report a new kind of seed dormancy and identify the storage behavior category for an important understory rainforest tree that also is used as an ornamental. While studying seed dormancy of Fabaceae species in Sri Lanka, we observed a considerable delay in emergence of the plumule following radicle emergence in Humboldtia laurifolia. Because epicotyl dormancy has not been reported in Fabaceae, we undertook a detailed morphological study of seed germination in this species. Our aims were to document desiccation tolerance/intolerance and epicotyl dormancy in seeds of H. laurifolia. Drying and low temperature storage were used to evaluate storage behavior of the seeds and imbibition, germination, and seed coat anatomy to categorize seed dormancy in two seed collections. Plumule development before its emergence and effects of light and temperature on plumule emergence were monitored. All seeds that were dried to 15% moisture content or stored at -1°C lost viability. Plumules began to grow 20 ± 5 d from radicle emergence and emerged after 40 ± 3 d. Dark and high illuminance further delayed plumule emergence. Seeds are recalcitrant and have a hitherto unreported kind of epicotyl dormancy, for which we propose the formula .     

Survival of Bradyrhizobium sp. (Arachis) on fungicide-treated peanut seed in relationship to plant growth and yield

Survival and viability of Bradyrhizobium inoculant on fungicide-treated peanut seed and the resulting effects on nitrogen fixation, plant growth and seed yield were determined. Vitavax and Benomyl had the most and least lethal actions against Bradyrhizobium strains grown on YEM medium containing a fungicide, respectively, while Thiram and Captan effects were intermediate. Survival of Bradyrhizobium USDA 3384 and USDA 3456, as single strain peat inoculants, on peanut (Arachis hypogaea L. var. Florunner) seeds treated with Benomyl or Vitavax at the rate of 3g/kg seed was also examined. Both fungicides inhibited the growth and affected the survival of strain USDA 3384 on peanut seed. Vitavax killed the inoculant in 9 h. In contrast, USDA 3456 resisted both fungicides, and survived for up to 72h. Nodule formation on greenhouse-grown plants inoculated with USDA 3384 was inhibited by all fungicides. Shoot dry weight and plant nitrogen content significantly decreased as compared to controls. Fungicides, except Vitavax, had a slight effect on nodulation and plant growth when USDA 3456 was used as inoculant. The agronomic importance of fungicide-inoculant interaction was examined in field experiments conducted in Egypt in soil free of peanut-nodulating Bradyrhizobium, where seeds were treated with a combination of two fungicides and a single strain peat inoculant of either USDA 3384 or USDA 3456. All fungicides decreased nodulation, nitrogen fixation, plant growth and seed yield, especially with USDA 3384 as inoculant. Fungicides inhibited viability and survival of Bradyrhizobium on peanut seeds which decreased nodule formation leading to reduced peanut seed yield.     

Improvement of biological control capacity of Paenibacillus polymyxa E681 by seed pelleting on sesame

Sesame is an important vegetable crop for the production of oil in Korea. The main obstacle of sesame cultivation is the occurrence of damping-off diseases and wilt caused by a complex of soil-borne pathogens in fields cultivated for two or more successive years. To protect sesame seedlings against these diseases, Paenibacillus polymyxa E681, a plant growth-promoting rhizobacterium (PGPR) previously shown to suppress disease incidence and promote growth on cucumber and pepper in the greenhouse and field experiments, was evaluated for its capacity for biological control and growth promotion in vitro and in situ. Seed treatment with strain E681 alone did not show consistent protection. Therefore, seed pelleting with strain E681 was attempted to increase the seed size and improve the stability and effectiveness of biocontrol capacity by strain E681. Through screening of pelleting materials, a combination of clay and vermiculite was selected for further experiments to enhance seed germination and root colonization of strain E681 on sesame. In greenhouse trials, formulations of strain E681 reduced disease incidence in disease-conducive soil. In the field, pelleting of sesame seeds with strain E681 significantly reduced pre- and post-emergence damping-off compared to the non-treated or pelleting alone controls; pelleting also promoted the plant growth and the grain yield. Furthermore, the efficacy of strain E681 for biological control and plant growth promotion was improved by sesame seed pelleting compared to the treatment with strain E681 alone. Hence, the application of strain E681 via seed pelleting offers potential to overcome some of the problems associated with successive years of sesame cultivation.     

Improvement of biological control capacity of Paenibacillus polymyxa E681 by seed pelleting on sesame

Sesame is an important vegetable crop for the production of oil in Korea. The main obstacle of sesame cultivation is the occurrence of damping-off diseases and wilt caused by a complex of soil-borne pathogens in fields cultivated for two or more successive years. To protect sesame seedlings against these diseases, Paenibacillus polymyxa E681, a plant growth-promoting rhizobacterium (PGPR) previously shown to suppress disease incidence and promote growth on cucumber and pepper in the greenhouse and field experiments, was evaluated for its capacity for biological control and growth promotion in vitro and in situ. Seed treatment with strain E681 alone did not show consistent protection. Therefore, seed pelleting with strain E681 was attempted to increase the seed size and improve the stability and effectiveness of biocontrol capacity by strain E681. Through screening of pelleting materials, a combination of clay and vermiculite was selected for further experiments to enhance seed germination and root colonization of strain E681 on sesame. In greenhouse trials, formulations of strain E681 reduced disease incidence in disease-conducive soil. In the field, pelleting of sesame seeds with strain E681 significantly reduced pre- and post-emergence damping-off compared to the non-treated or pelleting alone controls; pelleting also promoted the plant growth and the grain yield. Furthermore, the efficacy of strain E681 for biological control and plant growth promotion was improved by sesame seed pelleting compared to the treatment with strain E681 alone. Hence, the application of strain E681 via seed pelleting offers potential to overcome some of the problems associated with successive years of sesame cultivation.