微生物固体培养基凝固剂研究进展*

明胶是最早使用的固体培养基凝固剂,已逐渐被琼脂所代替。琼脂由于形成凝胶后透明度高、保水性好、无毒、不被微生物液化等优点,逐渐成为最常用的凝胶剂。后来,又发现无机硅胶、瓜尔胶、卡拉胶在某些情况下可用作凝固剂。近年来,兴起一种基于微生物快速检测的快速测试片,其所用凝固剂发展到黄原胶、刺槐豆角、聚丙烯酸系等。但新型凝固剂在使用过程中仍存在许多弊端,因此,从吸水和保水的机理出发对其研究和改性是一项重要的任务。     

琼脂固体培养基培养蜜环菌菌索总DNA的提取

野外采集的蜜环菌[Armillaria mellea(Vahl.ex Fr.)Quel]在提取DNA前需要分离获得纯化的菌丝体。常规液体培养获得菌丝团的方法感杂率较高,采集固体培养基表面cellophane膜上形成的菌丝则难以获得足量的DNA提取材料。蜜环菌细胞内含有大量多醣类物质,也使得蜜环菌高质量DNA的提取存在一定的困难。本研究通过改进试验,提供一个直接从琼脂固体培养基培养的蜜环菌菌索中提取高质量DNA的方法。其中样品的预先冻融处理方法可以促使蜜环菌菌索与琼脂分离;而在裂解提取缓冲液裂解材料细胞后加入1.25 mol/L KAc溶液,则有利于除去蜜环菌细胞内的多醣类物质以及残留的少量琼脂。通过琼脂糖电泳、紫外分光光度计对DNA浓度及OD值的测定、ISSR引物的PCR扩增以及酶切产物的PCR扩增等方法的检测,结果均表明该方法提取的DNA质量较好,符合进一步进行分子生物学研究的要求。     

标准化微生物培养基研究进展

标准化微生物培养基研究进展吴清平,陈素云,阙绍辉,丘万英（广东省微生物研究所,广州５１００７０）７０年代以来,微生物学技术方法已从传统技术方法的改进、完善和应用范围的扩展,逐步在微量、简易监测系统的兴起、发展和应用的基础上,发展到微生物的自动化快速诊...     

植物组织培养中琼脂浓度和pH对培养基凝固程度的影响

植物组织培养中多数植物pH要求5.0～6.2,琼脂浓度一般选用7～8mg·L-1,最高可达16 mg·L-1,较低的5 mg·L-1,而我们观察到培养基凝固程度受pH影响较大,适当的降低琼脂浓度不仅可以满足外植体的正常支撑需求,且培养效果更佳.本文研究pH和琼脂浓度对培养基凝固程度的影响,以确定组织培养技术中合理的琼脂浓度.     

培养基琼脂用量计算的商榷

植物组织培养中,大多数采用固体培养基。固体培养基是以琼脂(又称洋菜)为支持物的。目前一些资料中一般报告用培养基琼脂的百分含量来表示培养基的硬度。笔者认为,这种提法不太科学,而应该用胨力强度(又称凝胶强度)来表示琼脂培养基     

分枝杆菌全合成琼脂培养基的研究

本文报道了一种适合常见致病分枝杆菌生长的全合成固体琼脂培养基体系。它为实验室研究分枝杆菌的营养生理、生化特性、遗传变异、药理和耐药机制提供了一种新工具,也为研制更加满意的分枝杆菌培养基创造了条件。试验过的大多数非典型分枝杆菌,在全合成琼脂培养基上比在罗氏培养基上生长快,生长量少于或等于罗氏培养基。标准牛型株的细胞群体中,仅有少量细胞能在全合成琼脂培养基上生长。鸟型．胞内和偶发等分枝杆菌标准株在全合成琼脂培养基上产生两种不同的菌落。     

分枝杆菌简化琼脂培养基的研究

报道三种组分简单、制备方便的简化琼脂培养基.试验结果表明:7个标准菌株在简化琼脂培养基309A和309C上的生长速度和数量均相似或优于改良罗氏培养基和92号土豆汤琼脂培养基.简化琼脂培养基309C和309A可用于结核杆菌分离和药敏试验.     

细菌测试片的冷水可凝凝固剂初步研究和应用

微生物测试片具有携带方便、不用配制培养基的优点, 为一种新型的检测工具。测试片的技术关键是冷水可凝胶, 本文通过研究新的冷水可凝凝固剂(HKG)的保水性能、pH、强度、微生物利用及降解和抑菌效果, 再组装成细菌菌落总数测试片, 进行特异性和食品样品效果检验, 并与倾注平板法比对, 评价其作为微生物测试片凝固剂的可行性。结果显示HKG对16种微生物均无抑菌作用, pH为中性, 强度为340.31 g/cm2 ? 48.71 g/cm2; 具有良好的保水性能; HKG在短时期(5 d)内不支持微生物生长, 不被分解利用, 在测试片和倾注培养法中5种测试菌株均可达相同的灵敏度(P = 0.438 > 0.05), 各菌株菌落数(P = 0.442 > 0.05)和食品样品菌落数(P = 0.718 > 0.05)比较在统计学上无显著差异。结果表明, HKG能较好地应用于细菌测试片计数, 并能提高检测效率。     

培养基琼脂浓度及抗生素对3种海洋微藻生长的影响

目的:确定用于分离、纯化及保种所用的固体培养基中优化的琼脂浓度和利于微藻生长并抑菌的抗生素浓度。方法:设计固体培养基中的琼脂浓度以及培养液中抗生素的种类和浓度,定时测定培养体系中三种微藻的生物量及细菌菌落随培养时间的变化。结果:利于固体培养基上三种微藻生长的琼脂浓度均为0.6%~0.8%;牟氏角毛藻用200IU/ml的庆大霉素和200IU/ml的青霉素联合作用,球等鞭金藻8701和盐藻分别用300IU/ml和400IU/ml的青霉素时可保证微藻较好生长。结论:不同微藻生长所需的抗生素浓度不同,抗生素对微藻生长的影响不完全取决于抑菌作用,青霉素促进球等鞭金藻8701生长主要是产生促生长因子,庆大霉素抑制生长可能是产生抑制因子。     

Gelling agents for tissue culture of the seaweed Hizikia fusiformis

Callus and blade formation of the seaweed Hizikia fusiformis depended on the gelling agents used under axenic culture conditions. Excised cylindrical pieces (5 mm) of the hold fast were cultured on seven different gelling agents in seawater with added Provasoli's enrichment (PESI), at 40 μmol m⁻² s⁻¹ light intensity, 18 −C for 1 month. The highest percent of callus formation (47%), from holdfast pieces, was produced on solid medium composed of 2.0% high gel strength agar. No callus was formed in liquid medium. Blades, from holdfast pieces, were formed in PESI liquid medium at the rate of 45%, while the high level of axenic blade formation (30%) on solid support was observed on 0.5% high gel strength agar. Callus and blade were identified with the original strain, at the DNA level, using random amplified polymorphic DNAs. This revised version was published online in August 2006 with corrections to the Cover Date.     

Differential performance of marubakaido apple rootstock shoots grown in culture media containing different agar brands: dynamic rheological analysis

Shoots of the marubakaido apple rootstock grown in culture medium containing BBL agar presented significantly lower multiplication rate (MR) compared to MRs found for shoots grown in medium containing A-7002, A-7921, Select, and Phytagar as gelling agents. In addition, significant hyperhydricity was found for shoots grown in Phytagar and A-7921 agar-containing media. Analysis of elastic (G′) and viscous (G″) modulus showed that for all of the five agar brands used in this study, G′ was always much higher, i.e., typically one order of magnitude higher than G″, which characterizes a strong gel. G′ changed randomly with time for all of the agar brands studied, except for BBL, which presented progressive decline in G′ throughout the culture cycle. Examination of G′, within the same week, showed that Select agar always had the smallest G′, while Phytagar always had the highest G′. Analysis of the loss tangent (tan δ = G″/G′), a better indicator for gel behavior compared to G′ isolated, showed that tan δ for Select and Phytagar were always between tan δ values found for A-7002 and BBL. In addition, analysis of tan δ also indicated that BBL and Select agars showed a significantly weaker gel network, compared to Phytagar, A-7002 and A-7921 agars after the third week of culture. When seen together, these results indicate that shoot performance for the marubakaido rootstock is not related to agar gel strength. In addition, the high hyperhydricity rate found for shoots grown on agars A-7921 and Phytagar could not be related to agar gel strength, as well. Analysis of HPSEC profiles indicated that the best performance, i.e., multiplication rate, of marubakaido shoots in agars A-7002 and A-7921 is likely to be related to their lower polydispersity and/or smaller amount of high molecular weight fractions, compared to BBL, Phytagar, and Select agars.     

Selected Gelling Agents in Canned Dog Food Affect Nutrient Digestibilities and Fecal Characteristics of Ileal Cannulated Dogs

Little is known about the effects of gelling agents in canned dog food on nutrient digestibilities and fecal characteristics. Dogs were fed canned diets with either no gelling agent (control) or one of three gelling agents, wheat starch, a guar gum/carrageenan mixture (50:50), and a locust bean meal (LBM)/ carrageenan mixture (50:50) incorporated at two levels (0.2 and 0.5% of the diet on a wet weight basis). Six dogs were fed each diet in a 6 ‐ 7 Youden square design. Dogs fed diets containing gelling agents had higher ileal digestibilities of OM (P = 0.05), fat (P < 0.01), GE (P = 0.02), and total amino acids (P = 0.04) and lower (P < 0.01) total tract DM digestibilities when compared to dogs fed the control diet. Fecal output by dogs fed the control diet, expressed on both an as-is (73.4g/d) and DM (45.6g/d) basis, was lower (P < 0.01) than for dogs fed gelling agent-containing diets (mean 102.3g/d as-is and 57.6g/d DM). Dogs fed the control diet also had lower ( P < 0.01) fecal DM percentages and higher (P = 0.02) fecal scores than dogs fed gelling agent-containing diets.     

微生物培养基质量控制技术和标准

微生物培养基的酸碱度、凝胶强度和选择性等直接影响到培养基的质量,在理化试验方法中采用连接可渗透陶器型液体接头的电极和平头电极或者连接微型探头的电极可分别测定液体和固体培养基的pH值,而采用Gelometer和the LFRA Texture Analyser可测定固体培养基的凝胶强度。在微生物学方法中固体培养基采用倾注平板法、涂布法、划线法（半定量法）、改良的Miles-Misra法等测定生长情况,液体培养基采用稀释法测定生长率,用目标菌和杂菌的混合菌株评价选择性增菌培养基的选择性,利用OD值评价液体培养基生长率等。ICFMH（国际食品微生物学和卫生学委员会培养基工作组）、ISO、FDA以及我国卫生部等相继制定了培养基质量控制的标准,但目前还没有一个系统的适合我国国情的培养基质量控制国家标准,以致各相关单位采用的标准不一致,所以制定培养基质量控制国家标准非常关键。     

食品卫生微生物学检验培养基质量分析

为探讨培养基ISO国际标准对国内培养基质控的等效性,选择国内外4家培养基生产厂家的霉菌培养基、酵母菌培养基和显色培养基,从质控菌株生长率、抑菌效果等方面进行质量分析比对试验.生长率质控标准菌株在各种霉菌培养基、酵母菌培养基上的生长率均值均大于0.5,在显色培养基上的生长率均值均大于0.7,符合ISO国际标准.经统计分析,试验的4家培养基厂家生产的培养基质量水平差异不显著(P＞0.05),这为将新出台的培养基国家标准拟等效采用培养基ISO国际标准提供了基础材料和依据.     

微生物在多孔介质中渗流的数学模型

建立了一个完整的描述微生物在地层多孔介质中渗流的数学模型,模型中考虑了微生物的生长、营养物的消耗和微生物的存在对多孔介质物理性质的影响．此外,提供了该方程的求解过程和方法。实例计算结果表明,该方法能较好的描述微生物在多孔介质中的运移状况。     

Evaluation of kappa carrageenan as a substitute for agar in microbiological media

Seventy-one samples of the colloid kappacarrageenan extracted from 12 seaweed species were subjected to a number of standard physical demands of solid bacteriological culture media. All samples had a lower melting temperature (less than 67° C) than agar and a gelling (setting) temperature between 16° C and 51° C, some the same and others lower or higher than agar. Temperature spreads were narrow (ca 10° C) to broad (ca 30° C), depending on the seaweed source, but none were as broad as that of agar (ca 40° C). The majority of commercially prepared samples held a slant when incubated at 37° C, but California seaweed colloids were best at 28° C in this test. The majority of samples released little to no water of syneresis in slant tests as well as in plates. Some plates prepared with the colloid were crystal clear as compared to agar plates. All test microorganisms grew as well on kappa-carrageenan media as on agar media. Some media responses could be attributable to the seaweed species, but others could be traced to chemical extraction methods and modification of the colloid.     

GELRITE as a Gelling Agent in Media for the Growth of Thermophilic Microorganisms

GELRITE, a new gelling agent with good thermal stability and clarity, was evaluated in media for culturing selected thermophilic microorganisms. It was also evaluated for performing counts of thermophilic bacteria from soil samples. In most cases, GELRITE was shown to be superior to agar for these applications.     

Configurable Network Processing for Mobile Agents on the Internet

This paper presents a framework for building and deploying protocols for migrating mobile agents over the Internet. The framework enables network protocols for agent migration to be naturally implemented within mobile agents and then dynamically deployed at remote hosts by migrating the agents that perform the protocols. It is built on a hierarchical mobile agent system, called MobileSpaces, and several protocols for migrating agents for managing cluster computing systems have been designed and implemented based on the framework. This paper describes the framework and its prototype implementation, which uses Java as both the implementation language and the protocol development language.