观音土曲乳酸菌的分离与分子鉴定

[目的]指导白酒生产,提高白酒质量.[方法]采用平板分离法从观音土曲中分离到7株乳酸菌(B1～B7),测定其发酵产乳酸的量.选取乳酸产量最高的1株菌B7,提取其DNA并进行16S rDNA的PCR扩增、克隆、测序.从GenBank中选取与B7同源性最高的典型菌株,下载其16S rDNA序列,采用邻位相连法构建进化树.[结果] B1～B7的产乳酸量分别为:24.7、20.7、30.7、15.2、9.5、21.8、35.7 mg/100ml;对B7的基因组DNA进行16S rDNA的PCR扩增,并对纯化后的扩增产物进行TA克隆,阳性克隆经琼脂糖凝胶电泳后在3 000和1 600 bp处出现电泳条带,说明B7的16S rDNA PCR扩增产物已克隆到测序载体上.同源性分析结果表明,B7为乳杆菌属的短乳杆菌.[结论]该研究从观音土曲中分离到1株高产乳酸菌--短乳杆菌.     

饲料玉米中优质乳酸菌的分离和鉴定

[目的]从饲料玉米中筛选适合做青贮饲料添加剂的高效乳酸菌.[方法]用MRS+CaCO<,3>固体培养基筛选乳酸菌,经形态学和生理生化试验进行初步鉴定,测定其产酸效率;挑选出12株产酸率强的乳酸菌进行耐盐和耐酸能力的测定,并时其进行16 S rDNA 鉴定.[结果]从饲料玉米中共分离得到44 株乳酸菌,12株产酸能力强的菌株经生理生化试验鉴定分别属于明串珠菌属、乳杆菌属和肠球菌属,且12株菌株都具有良好的耐盐、耐酸能力.经16 S rDNA鉴定:A4、B9、B11、B12 和B14 为植物乳杆菌;A1、A2、A7、A11和B8为肠膜明串珠菌葡聚糖亚种;A8和A9为兼性肠球菌.[结论]从饲料玉米中筛选出具有较强产酸能力的乳酸菌,作为青贮饲料添加剂.     

1株出芽短梗霉的分离·纯化及鉴定

[目的]为出芽短梗霉的进一步开发利用提供理论依据.[方法]采用平板稀释法从土壤中分离出1株出芽短梗霉,用YPD培养基培养菌丝体,显微成像获得菌丝微观形态,利用分子生物学方法对其rRNA基因内转录区段(ITS 区)进行克隆测序,并与Genbank中已知序列进行比较.[结果]ITS区PCR扩增产物测序结果表明,所扩增序列的长度为606 bp,与Genbank中短梗霉的同源率为98%～99%,与Aureobasidium pullulans wb 149、A.pullulans HK58-1(2)属于同一单独分枝;形态及分子鉴定结果表明,出芽短梗霉培养成功.[结论]该研究采用经典与现代分子技术相结合的方法鉴定出了1株出芽短梗霉,为出芽短梗霉的分类鉴定提供了依据.     

耐受高氟环境生长的浸矿菌种的选育

通过对生物浸矿菌种耐氟能力的考察,驯化选育出具有较高耐氟能力的菌种,并得到了菌种耐氟生长的最佳条件。摇瓶试验结果显示,在所考察的4种菌种(代号DX,BS,ZJ,SG)中SG菌耐氟能力最强。通过影响因素试验发现,在初始p H值2.5、接种量40%、摇床转数200 r·min-1、温度33℃的条件下,SG菌耐受F-浓度能达到1000 mg·L-1,且完全氧化9.0 g·L-1Fe2+只需要24 h。收集SG菌耐氟浓度为0和1000 mg·L-1生长达到旺盛期时的菌体,在扫描电镜(SEM)下观察发现耐受氟驯化前后菌体形态变化较小,说明通过较长时间的驯化,SG菌能适应高氟浓度环境生长,并能保持较好的细菌氧化活性,是一株优良的耐氟菌株。运用16S rDNA克隆文库技术研究了浸矿菌种的种群组成,结果表明耐氟能力最强的SG菌的种群组成单一,与Acidithiobacillus ferrivorans菌相似度达到99%。     

嗜酸氧化亚铁硫杆菌的分离鉴定及对废胎面胶的脱硫再生

从河北兴隆某硫铁矿筛选到一株硫杆菌YT-1,经理化性能和16S rDNA序列分析鉴定为嗜酸氧化亚铁硫杆菌(At.ferrooxidans YT-1),研究了该菌对废胎面胶(GTR)的脱硫再生作用.研究结果表明,三元共混物丁苯橡胶(SBR)/炭黑(CB)/再生GTR比SBR/CB/未再生GTR拉伸强度和断裂伸长率分别提高了5.3%和11.1%;X射线光电能谱检测表明SBR/CB/再生GTR表面硫元素质量分数降低13.92%,且峰位往高氧化态偏移0.2 eV,说明再生GTR表面硫元素被氧化;共混胶断裂面扫描电子显微镜显示再生GTR与SBR/CB基体粘合性能显著提高,说明再生GTR表面硫交联键被打断.     

BIOLOG自动微生物鉴定系统在水产动物病原菌检测中的应用

[目的]了解BIOLOG自动微生物鉴定系统对水产动物病原菌鉴定结果的可靠性,为水产动物细菌性疾病的诊断提供参考.[方法]选用BIOLOG自动微生物鉴定系统对9株分离自发病水产动物的病原菌进行鉴定,同时采用16S rDNA序列分析对其中的4株进行鉴定,比较2种鉴定结果的符合率.[结果]采用自动微生物鉴定系统对9株病原菌进行鉴定均取得良好结果,采用16S rDNA序列分析对4株病原菌进行鉴定的结果与应用自动微生物鏊定系统所得结果完全一致,二者符合率为100%.[结论]应用BIOLOG自动微生物鉴定系统对水产动物病原菌进行鉴定具有操作简便、快速、准确等特点,是实验室鉴定水产动物病原菌的一种可行方法.     

牙鲆鱼肠道鼠李糖乳杆菌P15的分子鉴定与系统发育分析

[目的]对1株分离自健康牙鲆鱼肠道固有菌群的乳杆菌P15进行分子鉴定.[方法]利用PCR技术测定该菌株的16S rRNA基因序列,然后在GenBank中进行相关细菌相应序列的同源性比对,利用MEGA(4.0)软件进行系统发育学分析.[结果]获得了该菌株的16S rRNA基因序列(登录号为 AY852248).菌株P15与鼠李糖乳杆菌(Lactobacillus rhamnosus)的亲缘关系最近.[结论]菌株P15被鉴定为鼠李糖乳杆菌.     

多环芳烃厌氧降解菌的筛选与鉴定

[目的]筛选并鉴定多环芳烃厌氧降解菌.[方法]通过富集,在厌氧条件下从受焦油长期污染的土壤中筛选出多环芳烃的高效厌氧降解菌,并对其进行了生理生化试验和16S rDNA鉴定.[结果]从受焦油长期污染的土壤中分离出2株多环芳烃降解菌W2和Y3,经综合表征和16S rDNA序列分析,初步鉴定菌株W2为鞘氨醇单胞菌属(Sphingomo sp.),菌株Y3为芽孢杆菌属(Bacillus sp.).[结论]为多环芳烃的生物降解研究提供了理论依据.     

角蛋白分解真菌的分离筛选及初步鉴定

[目的]为动物角蛋白的高效、经济降解和利用提供新种质资源.[方法]采用传统分离筛选、固体基质降解和动物试验相结合的方法,对角蛋白降解菌进行了分离筛选及初步鉴定.[结果]得到1株非致病、降解效率较高的丝状真菌,并初步鉴定为毛霉科(Mucora-ceae)的一个种.[结论]该菌株的发现丰富了角蛋白降解菌的家族成员,为动物角蛋白的降解利用提供了新种质资源.     

一株降解芘的苍白杆菌的分离、鉴定及性能表征

采用富集培养的方法从北京焦化厂多环芳烃(PAHs)污染土壤中筛选到一株高效降解芘的微生物,命名为PW,分子生物学等手段鉴定此菌株属于苍白杆菌属(Ochrobactrum sp.).经一次性大剂量方法对此菌株进行驯化后,考察了摇瓶条件下环境因素对此菌株降解芘效率的影响.结果表明,驯化培养使得菌株5d内对0.5mmol/L芘的降解率由62.3%提高到92.7%.此外,该菌株的环境耐受性好,在环境温度为20￣40℃下该菌株对芘均具有一定的降解能力,30℃培养时降解效果最好;在pH为5～10的培养基中,PW对芘的降解率均在45%以上;当盐度小于3%时,此菌株对芘降解率在60%以上;同时菌株PW还可耐受一定浓度的重金属.     

桃根癌病菌拮抗放线菌抑菌活性物质的分离及其初步鉴定

[目的]分离并初步鉴定从桃树高发根癌病土壤中获得的拮抗线菌G19菌株的抑菌活性物质.[方法]采用蛋白沉淀法对拮抗放线菌G19菌株的抑菌活性物质进行粗提,利用高效液相色谱仪、中压制备色谱仪对其进行分离、纯化,应用MALDI-TOFMS法进行分子量的测定,最后通过化学显色反应进行相关官能团的验证.[结果]经分离纯化后拮抗放线菌G19的抑菌物质被抨击出7个肽段,是分子量范围为900～1 300 Da,同时推断其含有一个Cys并携带H<,2>O,Na<'+>;官能团显色反应验证该物质为多肽且含有糖基.[结论]研究结果为拮抗放线菌G19菌株的抑菌物质结构的最终确定奠定了基础.     

Epidemiology of S. aureus cross-infections in dermatological wards

Over a three-year period, 196 of 3115 patients admitted in a dermatological department became infected with S. aureus (6,2 %). 205 strains of S. aureus were isolated. Serologic typing, phage-typing and antibiotic sensitivity tests revealed 3 epidemic and 2 endemic strains. The 3 epidemic strains infected 24 patients: 12 from july to november 1972 were infected with a serotype 66438 S. aureus resistant to fusidic acid. 6 patients (male) were infected with a serotype III in february and march 1972; 8 patients were contaminated with a serotype 18 S. aureus from december 1973 to february 1974, after staying in a surgical department. Of the 2 endemic strains 1, phage-pattern 53/79, is non-typable by serologic-typing; this strain has been observed only in the dermatological department and 20 patients were infected with, from january to october 1974. The second endemic strain, phage-pattern 81/+ serotype I, cross-infected 16 patients during this three-year survey; 12 of them were admitted repeatedly. During this three-year survey, it could be proved that, at least, 1 out of 3 patients is infected with an epidemic or an endemic strain. We can suggest that the factors enhancing cross-infection in dermatological department are: the sex of patients (80 % were male); presence of a tween splitting enzyme by S. aureus promotes growth of Staphylococci on the skin; patients transfered from a department to another or repeatedly admitted are more often infected. But, as they are source of some outbreaks, they need special measures (asepsis and hygiene); cortico?ds or immunodepressors enhance cross-infection; antibiotics must not be only limited but varied too.     

Isolation and structure flucidation of alkaloids from the bulb of Fritillaria Wabuensis S.Y. Tang et S.C. Yueh

A versatile photochemical method of labeling human antibodies is described. Labeling is achieved by photolyzing 4-azido-2,3,5,6-tetrafluoro-14C-methylbenzoate and the B72.3 human antibody in a buffer at physiological pH. The photochemically produced nitrene presumably inserts into bonds in the hydrophobic part of the antibody resulting in > 75% attachment of the photoprobe. An immunoassay of B72.3 with mucin (B72.3 antigen) reveals > 97% retention of immunoreactivity and suggests that photochemical labeling is a viable alternative for the conjugation of biomolecules.     

马铃薯炭疽病菌分离与鉴定

[目的]对进境船舶上携带的带病马铃薯薯块进行了病原真菌的分离鉴定,明确其检疫重要性,并对病菌的快速鉴定方法进行研究.[方法]通过形态学观察和ITS序列分析对病菌进行了鉴定,并对已有的巢式PCR方法进行了验证.[结果]形态学观察和ITS序列分析表明,病原菌为球炭疽菌(Colletotrichum coccodes).巢式PCR中用特异性引物Cc1NF1/Cc2NR1扩增 C.coccodes得到了349 bp特异性条带.[结论]巢式PCR方法可以用来对C.coccodes进行快速鉴定,该病原菌是进境植物检疫潜在危险性病原真菌,在国内口岸属首次截获.     

高效盖草能降解菌的分离与鉴定

针对海南地区长残效除草剂残留对后茬产生药害问题,以高效盖草能作为研究对象,采用富集培养的方法从长期施用长残效除草剂的土壤中分离到3株细菌,经过对这3株降解菌的培养特性及生理生化特征的测定试验,经传统鉴定方法对3种菌种进行鉴定,结果显示,HM1和HM2同为芽孢杆菌属(Bacillus sp.),HM3为甲基球菌属(Methyloeoccus sp.).并分别研究了培养含糖量、温度、pH和高效盖草能初始浓度对分离出的3株降解菌生长的影响,结果表明,HM1和HM3的最适含糖量是0.HM2的最适含糖量为0.5%;HM1和HM3的最适温度范围相同,为20～37℃,HM2的最造温度为25～41℃;HM1、HM2和HM3的最适生长温度分别是35、30和30℃;HM3的生长对酸碱度要求不严格,最适生长pH范围为6.0～9.0.HM1适宜在中性偏酸的条件下生长,而HM2则适宜在中性偏碱的条件下生长,其最适生长pH均为6.0～8.0;HM1和HM2生长的最适高效盖草能浓度是100 mg/L,HM3生长的最适高效盖草能浓度为200mg/L.     

Molecular Mechanism of Granulation. II: Proton Translocating Activity

A laboratory-scale upflow anaerobic sludge blanket (UASB) reactor was used in this study to produce granular sludge at mesophilic temperatures (35 ± 1°C). After more than 150 days of operation, a COD removal efficiency of 95% was achieved with an organic loading rate of 8.73 gCOD∕L∕day. At the same time, the sludge granulation process was observed. The mature granules were examined for their stability in terms of the presence of calcium ion, surfactant, pH (buffer and H2SO4∕NaOH solution), metabolic inhibitor (iodoacetic acid and sodium fluoride), and proton translocator (carbonyl cyanide m-chlorophenyl-hydrazone). The results showed that bacterial surface dehydration, biological metabolic activity, and proton translocating activity were directly related to the strength of UASB granules. This indicated that the proton translocating activity on bacterial surfaces was the crucial factor in sludge granulation and, as a consequence, supported the proton translocation-dehydration theory. Experimental results from other studies were also used to support this new theory.     

耐盐菌的筛选及初步鉴定

[目的]筛选耐盐菌株,为含盐废水的处理提供支持.[方法]以某化工有限公司生产污水为菌株来源,从中分离纯化得到耐盐菌株,通过测定所筛选得到的不同菌株的耐盐度,把不同的菌株接种到生产废水中,摇床培养2d后测COD,通过计算COD去除率确定一株优势菌株,并将其进行基本的生理生化特征测试.[结果]初步鉴定该菌为一株革兰氏阳性细菌,最适生长温度为37℃,最适生长pH值为7.0,NaCI盐浓度为1%时生长情况最好,经驯化培养耐盐度达9%.[结论]该菌株为合格的耐盐菌.