对生态系统中分离获得的菌种进行优势种群测定的方法学…

两年内７次从同一个以杨木木屑为原料的厌氧发酵罐的高稀释度发酵淮中分离出一株中温纤维分解菌，即新种阿氏梭状芽孢杆菌。从细菌计数高且能反复分离获得这两个特点，可以断 这下阿氏梭状芽孢杆菌在该生态系统中曾是一个优势种群。经菌落及菌体形态学观察，以及用首次分离的菌株所作的多价抗血清酶联免疫吸附试验（ＥＬＩＳＡ），确定了７次分离菌株为同一菌种，从而证实阿氏梭状芽孢杆菌在１９８６年９月至１９８８年８月间，曾是     

绿色荧光蛋白基因标记内生枯草芽孢杆菌

用枯草芽孢杆菌168菌株rpsD基因的启动子替换质粒pGFP4412中蜡状芽孢杆菌4412启动子,从而构建了能在枯草芽孢杆菌中表达绿色荧光蛋白基因gfpmut3a的载体pS4GFP,将其导入具有内生、防病、促生作用的野生型枯草芽孢杆菌BS-2菌株中,筛选获得遗传稳定性好且具有良好发光表型的标记菌株BS-2-gfp.该标记菌株在小白菜体内的定殖研究结果表明,该菌株能够在小白菜根际及根、茎、叶内定殖和传导,接菌50d后仍能在其体内分离到标记菌株.图5参17     

一株产胞外多糖的山药内生细菌Lysinibacillus fusiformis S-1的分离和鉴定

微生物的胞外多糖是重要的生物资源,为获得新型的具有药用价值的胞外多糖产生菌,从山药、地瓜、马铃薯和胡萝卜的根茎组织中分离、筛选到11株能产胞外多糖的植物内生菌,利用苯酚–硫酸法对这11株菌的多糖产量进行了定量分析.对多糖产量最高的S-1菌株,检测其在发酵过程中菌体生长、胞外多糖生成以及发酵过程的pH变化,绘制其胞外多糖发酵代谢曲线.通过形态观察、培养特性观察、生理生化实验和16S rDNA序列分析对该菌株进行了鉴定.结果显示,S-1菌株在产糖培养基中可以产生1.50 g/L的胞外多糖,在11株菌中产量最高.16S rDNA序列分析显示该菌属于赖氨酸芽孢杆菌(Lysinibacillus),且与纺锤形赖氨酸芽孢杆菌(L.fusiformis)亲缘最近.综合其形态特征、培养特性和生理生化实验结果,将S-1菌株鉴定为纺锤形赖氨酸芽孢杆菌.该菌在胞外多糖产生菌中少有报道,为胞外多糖的进一步研究提供了菌种资源.     

云南,西藏热泉中主要微生物类群及其金成矿作用的研究

从地质微生物生态学的角度研究和阐述了云南和西藏热泉中主要微生物类群及其金成矿作用．发现了热泉中存在的嗜热好氧菌优势菌属为并孢杆菌（Racillus），放线菌中的优势菌属为链霉菌（Streptomyces），真菌中优势菌属为曲霉（Aspergillus），厌氧菌中优势菌为硫酸盐还原菌的脱硫肠状菌属（Desulfotomaculum）和核状芽孢杆菌属（Clostridium），对吸附聚金、沉淀金的功能菌进行的研究发现．嗜热细菌对溶解金有明显的吸附聚金作用，表明原溶液金大多富集于细胞体；并以芽孢杆菌属K12－1菌株等进行了吸附聚金条件试验．     

乐果降解菌LGX1的筛选及其降解特性研究

通过富集培养,从连续施用农药乐果的土壤中分离得到一株具有较强降解有机磷农药乐果能力的细菌菌株LGX1,通过菌落形态观察及细菌的16SrDNA测序,对其进行了鉴定,同时初步研究了其降解性能。结果表明：该菌株为蜡状芽孢杆菌（Bacillus cereus）,在接种量为20%,乐果初始质量浓度为100mg·L-1,外加碳源葡萄糖质量分数为2%,温度为35℃,初始pH值为4时该菌株对乐果的降解能力最强;并且能在以辛硫磷、毒死蜱和三唑磷为唯一碳源的基础盐培养基中生长,且均比在以乐果为唯一碳源的基础盐培养基中生长的OD600值要高,初步推断菌株LGX1对有机磷农药的降解有一定的广谱性。     

硅酸盐细菌解钾兼拮抗活性菌株的筛选

从紫色土壤中分离筛选出具有解钾拮抗双重活性的硅酸盐细菌CS1和CS29菌株。接菌处理，其水溶性钾分别增加75％和65％，转化率达0．82％和0．72％。两株菌对革兰氏阳性的蜡样芽孢杆菌，金黄色葡萄球菌，藤黄八叠球菌等和革兰氏阴性的大肠杆菌均有较强的抑菌活性。筛选菌株中91．2％具有解钾活性，但拮抗活性菌株只占总数的11．6％，表明二性状之间无相关关系。表4参7     

一株中度嗜盐硫酸盐还原菌的分离及生理特性研究

本文从四川自贡盐厂卤水污泥中，分离出一株中度嗜盐硫酸盐还原菌。菌体为短杆状，０．９－１．１×２．０－２．５μｍ，革兰氏染色阴性，形成芽孢，不含脱硫孤菌素。固体培养基上无Ｆｅ２＋时，菌落为半透明状；有Ｆｅ２＋时菌落为黑色，直径为１－３ｍｍ。利用乳酸盐作为碳源，利用乙酸盐时必须添加生长因子。能在５％－２５％（Ｗ／Ｖ）Ｎａｃｌ浓度范围内生长，最适生长浓度为９％－１３％（Ｗ／Ｖ）。最适生长温度为３５℃。最适生长ｐＨ值为７．０．根据形态和生理生化特征，该菌为脱硫肠状菌属，但与该属已报导的硫酸盐还原菌在嗜盐、碳源利用、最适生长温度以及其它一些特征上均有不同，因此可能是个新种。该菌是目前已报道的嗜盐浓度最高的硫酸盐还原菌。     

垃圾堆肥微生物接种实验

从２２个垃圾堆肥、畜粪、土壤等样品中分离获得纤维分解菌１９８株，选其中２株生长快、粗纤维分解能力强的菌株制成菌剂，以０．０５％～０．１％的接种量加入二次发酵的垃圾堆肥中。结果，接菌堆肥比不接菌堆肥升温快且高，高温维持时间长，真菌和纤维分解菌数量多，腐植质含量提高２１％～２６％。肥效试验证明，施接菌堆肥比施不接菌堆肥的可使青莱增产９．９％。     

利用枯草芽孢杆菌株间原生质体融合进行差异表型遗传重组分析的研究

枯草芽孢杆菌D100和枯草芽孢菌TN179同为枯草芽孢杆菌Ki－2－132的两个不同菌株，具有明显的表型差异．它们的融合子D279兼有两亲本的遗传性状，能够在常规培养条件下稳定遗传．但经过EMS处理后，即产生形状与各自亲本相似，且伴随菌落颜色、菌体形状和一些生理变化的分离物．从这些分离物的表型改变，得出了基因及连锁的一些遗传值息．     

有机磷农药降解菌的紫外诱变育种

有机磷农药降解菌地衣芽孢杆菌（ Ｂａｃｉｌｌｕｓｌｉｃｈｅｎｉｆｏｒｍｉｓ） 经紫外线诱变后,筛选出突变菌株Ｐ１２ ．在θ＝３０℃,溶解氧ρ（Ｏ２） ＝２ ．５ ｍｇ Ｌ－１ 的培养条件下,３ ｄ 内对甲胺磷的降解率为８０ ．１％ ,比出发菌株提高了将近１０％的降解率．农药斜面连续传代１０ 次,降解活力保持稳定．     

The enumeration and characterization of bacteria and fungi associated with marine wood-boring isopods,and the ability of these microorganisms to digest cellulose and wood

Bacteria and fungi associated with the marine wood-boring isopods Limnoria lignorum and Sphaeroma serratum, and with their wood burrows and seawater were investigated. Plate counts using nutrient agar and Czapex Dox agar media in natural and in artificial seawater were used. The diversity and number of bacteria associated with the two isopods were much higher than those associated with wood or seawater, whereas the opposite was true for fungi. The hydrolytic activity of the predominant genera on carboxymethylcellulose and on different types of wood was investigated. The magnitude of fungal cellulase activity exceeded that of bacteria. Activities of bacterial extracellulases exceeded those of intracellulases, while the opposite was observed for fungi. Cellulolytic activity of the predominant isolates biodegraded labkh, peach, white oak and mahogany in decreasing order of activity. Evidence provided by the cellulolytic activities, detected in woodcontaining cultures, suggests that the predominant micro-organisms isolated may play an important role in the woodboring process. Specimens used in the present study were collected from Port Fouad at Port Said Harbour, Egypt.     

一株生物素依赖型菌株bio-5:分离,初步鉴定及其用于生物素浓度测定

从西红柿根圈土壤中分离纯化到一株生物素依赖型菌株bio - 5 ,经测定其内源抗生素抗性和脂肪酸特征 ,初步定名为假单胞菌bio - 5 .这一菌株对生物素非常敏感 .建立了以此菌株用于测定生物素浓度的标准方法 .利用这一方法 ,测定了不同作物叶子中生物素的含量 ,并将这一方法成功用于筛选转stv基因烟草植物 .与传统生物素测定菌株LactobacillusplantarumATCC80 14比较 ,我们所分离的菌株具有易于获得 ,易于培养 ,快速用于生物素浓度测定的特点 .图 7表 2参 2 2     

Increasing the performance of anaerobic digestion: Pilot scale experimental study for thermal hydrolysis of mixed sludge

The performance of a pilot plant operation combining thermal hydrolysis (170°C, 30 min) and anaerobic digestion (AD) was studied, determining the main properties for samples of fresh mixed sludge, hydrolyzed sludge, and digested sludge, in order to quantify the thermal pretreatment performance (disintegration, solubilisation, and dewaterability) and its impact on the anaerobic digestion performance (biodegradability, volatile solids reduction, and digester rheology) and end product characteristics (dewaterability, sanitation, organic and nitrogen content). The disintegration achieved during the thermal treatment enhances the sludge centrifugation, allowing a 70% higher total solids concentration in the feed to anaerobic digestion. The digestion of this sludge generates 40% more biogas in half the time, due to the higher solids removal compared to a conventional digester. The waste generated can be dewatered by centrifugation to 7% dry solids without polymer addition, and is pathogen free.     

猪场废水厌氧消化过程中的除磷效果

采用序批式半连续厌氧消化试验方法,研究猪场废水厌氧消化过程中磷的去除情况。结果表明,水力停留时间（HRT）为1、3、6和9 d的厌氧反应器平均除磷率分别为65.0%、81.1%、82.7%和83.0%,而COD平均去除率分别为54.5%、82.3%、87.0%和85.9%。厌氧反应器除磷能力随沼气产量的增加而增加,说明厌氧反应器中磷的去除与产甲烷过程密切相关。对厌氧消化前后的污泥进行浸提后发现,厌氧消化过程中,化学反应生成磷酸盐沉淀的除磷作用十分显著,污泥中正磷酸盐,与铁结合的磷化合物（Fe-RP）,还原可溶性磷,与钙、镁离子结合的磷化合物（Ca-RP、Mg-RP）以及无机或有机聚合磷增加量分别为0.027 8～0.101 5、0.013 5～0.081 0、0.2165～0.430 5、23.4～54.8和7.2～21.5 mg.g-1;且总体而言,HRT越长,污泥中与不同金属结合的磷增加量就越大。从HRT、磷和有机物的去除效果以及沼气产气速率3个方面综合考虑,猪场废水厌氧消化反应器的HRT控制在3 d为宜。     

辽北四位一体庭院生态系统冬季增温保温效应研究

观测表明，辽北庭院生态系统中日光温室、养猪暖舍、厕所、沼气池四位一体模式具有较好的增温保温效应，效果明显优于单一日光温室，可创造良好的养殖业环境，保证模式中沼气池在冬季良好运行。     

南海莺琼盆地沉积环境中厌氧纤维素分解菌的分布与作用

在严格厌氧条件下，研究了驾琼盆地东方１－１－１井垂直剖面不同沉积层地质样品中的厌氧纤维素分解菌．结果表明：厌氧纤维素分解菌在各样品中都存在，但其分布有很大随机性，与样品埋藏深度无相关性，与样品中一些有机物含量，如纤维素、半纤维素也无相关性，厌氧纤维素分解菌数量高的样品，其在生物模拟产甲烷实验中的单位有机质产气量也较高，有显著的相关性．菌体形态主要为杆菌和球菌．     

烟区烟草根际与非根际土壤微生物的生态因子作用分析

烟草根际和非根际土壤的理化性质、微生物区系和土壤酶之间的相互关系尚缺乏系统研究。对南阳烟区6种土壤包括黄褐土和黄棕壤等共13个取样点的烟草根际和非根际土壤的分析表明,根际与非根际微生物之间没有对应相关关系。根际土壤真菌数与解钾菌数、硝化细菌数显著正相关;非根际土壤荧光假单胞菌数与细菌芽孢数极显著正相关,氨化细菌数与放线菌数显著负相关。与土壤容重和空隙度显著相关的仅有根际真菌数(正相关)。与土壤全氮呈显著正相关的分别有根际荧光假单胞菌、纤维素分解菌(正相关)以及根际解磷菌和非根际真菌(负相关)。土壤全C与根际细菌总数显著负相关。土壤pH与非根际纤维素分解菌显著正相关。速效P与非根际放线菌数呈显著正相关。根际土壤蛋白酶活性与根际细菌总数极显著负相关。非根际土壤蛋白酶活性则分别与非根际放线菌和解钾菌显著正相关。根际土壤脲酶活性分别与根际真菌、解磷菌显著正相关,非根际脲酶活性则与非根际纤维分解菌显著正相关。非根际土壤过氧化氢酶活性与非根际细菌总数显著正相关。     

Killing of marine phytoplankton by a gliding bacterium Cytophaga sp., isolated from the coastal sea of Japan

A marine gliding bacterium Cytophaga sp. (strain J18/M01) was isolated from Harima-Nada, eastern Seto Inland Sea, Japan in 1990. This bacterium preys upon various species of marine phytoplankton. All of the five raphidophycean flagellates, all of the four diatoms, and one of the two dinoflagellates examined were killed within a few days when cultured with the bacterium. The bacterium presumably achieves this by direct attack, because the culture filtrate in which host organisms were totally destroyed had no significant effects on the growth of the same host organism (Chattonella antiqua). If one or a few bacterial cells were inoculated into C. antiqua culture, all of the host organisms were killed. The bacterium proliferated in filter-sterilized seawater, suggesting its ubiquitous existence in the coastal sea. The killing of phytoplankton by bacteria such as Cytophaga sp. J18/M01 may be a significant factor influencing the population dynamics of phytoplankton in nature and may contribute to the sudden disappearance of red tides in the coastal sea. Bacterial destruction of phytoplankton may also be a factor that regulates primary productivity in marine ecosystems.     

睾丸酮丛毛单胞菌teiR基因插入失活突变体构建及降解特性分析半

睾丸酮丛毛单胞菌(Comamonas testosteroni)是革兰氏阴性菌,能够以各种类固醇化合物作为唯一碳源和能源,生物信息学分析发现睾丸酮丛毛单胞菌teiR基因编码蛋白属于LuxR-type转录调控蛋白.利用同源重组原理构建teiR基因插入失活突变体C. mut,初步研究TteiR基因在类同醇代谢途径中的作用,以及对睾丸酮丛毛单胞菌关键降解酶3a-HSD/CR的表达影响.结果发现,teiR基因插入失活突变体C.mut在LB培养基中的生长速率不受影响,突变体C.mut抑制了teiR基因和关键降解酶3a-HSD/CR的表达,同时突变体C. mut丧失了以睾丸酮作为碳源进行生长的能力.结果表明teiR基因调节睾丸酮丛毛单胞菌类因醇代谢,同时teiR基因表达对3a-HSD/CR基因的表达是必需的.图7参14     

Biodegradation of paclobutrazol by a microbial consortium isolated from industrially contaminated sediment

Biodegradability of the plant growth retardant paclobutrazol by a microbial consortium in which Pseudomonas was the predominant strain was investigated in batch culture. The consortium which had been isolated from an industrially contaminated sediment was proven to be useful for the treatment of effluents containing paclobutrazol. Paclobutrazol was degraded by the pure isolated strain of Pseudomonas sp. as well as the microbial consortium. Paclobutrazol was utilized as the sole source of carbon and energy. Sixty percent of the paclobutrazol was degraded by the microbial consortium from an initial concentration of 54 mg L^?1 within 48 h and more than 98% of an initial concentration of 3.4 mg L^?1 was degraded within 36 h. The optimum temperature and pH were determined to be 30°C and 7.0, respectively. A pure strain of a bacterium, isolated from the enrichment culture was identified as Pseudomonas sp. The microbial consortium was tolerant of high pH and could degrade paclobutrazol faster than the pure strain. The degradation rate of this plant growth regulator in an aerobic environment was greater than that under anaerobic conditions.