以固氮菌和解磷菌筛选拮抗作物病害的细菌组合

用选择性培养基从土壤分离筛选了22 种具有固N或解P能力的细菌,不进行纯化,分组混合接种于15种液体培养基,进行继代培养,每代培养48 h.第12代时,各细菌组合的菌类数目及比例达到稳定状态,共获得15个细菌组合,各组合均含有2～3种细菌.对茄黄萎病病原菌(Verticillium dahliae)、棉花立枯病病原菌(Rhizoctonia solani)和黄瓜角斑病病原菌(Pseudomonas syringae)等9种植物病原菌进行平皿抑菌试验,筛选出7个具有抑菌作用的细菌组合,进一步从植株盆栽试验中筛选出6个有防病作用的组合,其中C₇和C₁₅表现出最明显的抑菌防病效果.C₁₅保持了固N能力,C₄、C₅、C₇、C₉、C₁₄和C₁₅保持了解P能力.结果表明,这是一条可获得具有协同作用的、稳定的多功能生防菌剂的途径.     

Decolorization of Direct Red 28 by mixed bacterial culture in an up-flow immobilized bioreactor

Aerobic mixed bacterial culture comprised of five isolates (Bacillus vallismortis, B. pumilus, B. cereus, B. subtilis and B. megaterium) identified by 16srDNA analysis was developed from wastewater samples from the aeration tank of an effluent treatment plant of a textile and dyeing industry and evaluated for its ability to decolorize azo dye Direct Red 28 in an up-flow immobilized packed bed bioreactor using marble chips as support matrix. The bioreactor was operated under two parameters: an aeration rate of 0.4 and 0.6 mmol/min at a flow rate of 60, 90 and 120 ml/h, respectively. At a constant aeration rate of 0.4 mmol/min and with flow rates of 60, 90 and 120 ml/h, optimum decolorization of 91, 75 and 72% was observed, while at an aeration rate of 0.6 mmol/min and flow rates of 60, 90 and 120 ml/h, optimum decolorization of 93, 78 and 72% was observed over 10 days. The study concluded that across the two aeration rates and the respective flow rates, the higher aeration rate of 0.6 mmol/min along with a flow rate of 60 ml/h was best suited to decolorize Direct Red 28 in the packed bed bioreactor. Spectral changes of the input and output of the bioreactor by UV–visible spectroscopy indicated decolorization of the dye solution by degradation in addition to the visual observation of the biosorption process.     

Biodegradation kinetics of 2,4,6-Trichlorophenol by an acclimated mixed microbial culture under aerobic conditions

The objective of this study was to achieve a better quantitative understanding of the kinetics of 2,4,6-trichlorophenol (TCP) biodegradation by an acclimated mixed microbial culture. An aerobic mixed microbial culture, obtained from the aeration basin of the wastewater treatment plant, was acclimated in shake flasks utilizing various combinations of 2,4,6-TCP (25–100 mg l⁻¹), phenol (300 mg l⁻¹) and glycerol (2.5 mg l⁻¹) as substrates. Complete primary TCP degradation and a corresponding stoichiometric release of chloride ion were observed by HPLC and IEC analytical techniques, respectively. The acclimated cultures were then used as an inoculum for bench scale experiments in a 4 l stirred-tank reactor (STR) with 2,4,6-TCP as the sole carbon/energy (C/E) source. The phenol acclimated mixed microbial culture consisted of primarily Gram positive and negative rods and was capable of degrading 2,4,6-TCP completely. None of the predicted intermediate compounds were detected by gas chromatography in the cell cytoplasm or supernatant. Based on the disappearance of 2,4,6-TCP, degradation was well modelled by zero-order kinetics which was also consistent with the observed oxygen consumption. Biodegradation rates were compared for four operating conditions including two different initial 2,4,6-TCP concentrations and two different initial biomass concentrations. While the specific rate constant was not dependent on the initial 2,4,6-TCP concentration, it did depend on the initial biomass concentration (X _init). A lower biomass concentration gave a much higher zero-order specific degradation rate. This behaviour was attributed to a lower average biomass age or cell retention time (θ_x) for these cultures. The implications of this investigation are important for determining and predicting the potential risks associated with TCP, its degradation in the natural environment or the engineering implications for ex situ treatment of contaminated ground water or soil.     

Investigation of the biotransformation of pentachlorophenol and pulp paper mill effluent decolorisation by the bacterial strains in a mixed culture

Mixed culture of two bacterial strains Bacillus sp. and Serratia marcescens showed potential pentachlorophenol (PCP) degradation and decolorisation of pulp paper mill effluent. The physico-chemical quality of pulp paper mill effluent has been analyzed after 168 h incubation period degraded by mixed culture. The study revealed that it has decreased high load of BOD, COD, TS, TDS, TSS, sulphate, phosphate, total nitrogen, total phenols, metals and different salts (i.e. chloride, sodium, nitrate, potassium) at 168 h incubation period. PCP degradation in pulp paper mill effluent was confirmed by HPLC analysis. Mixed culture was found to degrade PCP up to (94%) present in pulp paper mill effluent with 1% glucose and 0.5% peptone (w/v) at 30 ± 1 °C, pH 8.0 ± 0.2 at 120 rpm in 168 h incubation period. The simultaneous release of chloride ion up to 1200 mg/l at 168 h emphasized the bacterial dechlorination in the medium. The pulp paper mill effluent degradation was also supported by decline in pH, AOX (absorbable organic halides), color, D.O., BOD, COD and PCP. The analysis of pulp paper mill effluent degradation products by GC–MS analysis revealed the formation of low molecular weight compound like 2-chlorophenol (RT = 3.8 min) and tetrachlorohydroquinone (RT = 11.86 min) from PCP extracted degraded sample. Further, mixed culture may be used for bioremediation of PCP containing pulp paper mill waste in the environment.     

Simultaneous biodegradation of pyridine and quinoline by two mixed bacterial strains

Experiments were conducted to provide data on the effectiveness of bioaugmentation in the removal of pyridine and quinoline from different wastewaters. A pyridine-degrading bacterial strain (Paracoccus sp. BW001) and a quinoline-degrading strain (Pseudomonas sp. BW003) were isolated from the activated sludge of a coking wastewater treatment plant. In this study, a consortium of these two bacterial strains was used as inoculum to simultaneously degrade pyridine and quinoline in three types of wastewaters: sterile synthetic, domestic, and industrial. In addition, variation of the bacterial community structures during degradation was monitored by denaturing gradient gel electrophoresis and amplicon length heterogeneity polymerase chain reaction techniques. The results of our experiments indicate that pyridine and quinoline can be removed efficiently using this inoculum but that the degradation process results in the production of ammonium as a by-product. Also, in the two actual wastewaters investigated, we observed that several autochthonous strains of bacteria in both the domestic and industrial wastewater were tolerant of pyridine and quinoline and grew rapidly.     

Biodegradation of polyvinyl alcohol by a mixed microbial culture

A mixed culture capable of degrading 1 g l⁻¹ polyvinyl alcohol (PVA) completely was screened from sludge samples at Pacific Textile Factory, Wuxi, China. This mixed culture had stronger capability of degrading PVA with low polymerization and high saponification than degrading PVA with high polymerization and low saponification. Inorganic nitrogen source was more suitable for the mixed culture to grow and degrade PVA than organic nitrogen source. Microorganisms and relative abundance of this mixed culture were explored by terminal restriction fragment length polymorphism (T-RFLP). Small PVA molecules were detected in cell extracts of the mixed culture. This indicated that PVA degradation in the mixed culture was in fact a combined action of extracellular and intracellular enzymes. Two strains producing extracellular PVA-degrading enzyme were isolated from the mixed culture. They could individually degrade PVA1799 with polymerization of 1700 from initial average molecular weight 112,981 to 98,827 Da and 84,803 Da, respectively. However, only small amount of PVA124 in polymerization of 400 could be degraded by these two strains.     

The degradation of biphenyl and chlorobiphenyls by mixed bacterial cultures

Abstract Pseudomonas sp. HV3 grows on naphthalene but not on biphenyl, as the sole source of carbon. When the cells of Pseudomonas sp. HV3 grown on naphthalene were shaken with biphenyl as the carbon source in a mineral salt solution, a yellow metabolite identified as the meta -cleavage product of biphenyl was excreted. The degradation of biphenyl stopped here, but was completed if either 2-methyl-4-chlorophenoxy acetic acid (MCPA)-degrading mixed culture or a Nocardia strain was added to the growth solution. Neither of these uses naphthalene or biphenyl as growth substrate. The mixed culture of Pseudomonas sp. HV3 and Nocardia sp. also degrades the commercial polychlorinated biphenyl (PCB) mixture Aroclor 1221. A yellow metabolite was likewise produced in the degradation, and sometimes two different peaks of the yellow metabolite were observed. The gas chromatography-mass spectrometry (GC-MS) analyses showed that 40–87% of Aroclor 1221 was degraded during an incubation time of 6–21 days. Chlorobenzoic acids were found as metabolites.     

Proteomic tracking and analysis of a bacterial mixed culture

To improve the understanding of microbial behaviors in communities, proteomic tracking, an approach for relative quantification of species-specific population dynamics of mixed cultures, was developed. Therefore, a bacterial mixed culture was analyzed during batch cultivations with and without addition of the antibiotic Ceftazidime. The community was composed of Burkholderia cepacia, Pseudomonas aeruginosa, and Staphylococcus aureus, pathogens causing infections in cystic fibrosis patients. Gel-based proteomics and mass spectrometry were used to obtain qualitative and quantitative proteomic data. During cultivation, P. aeruginosa became dominant within the mixed culture while S. aureus was inhibited in growth. Analysis of samples - taken along cultivation - revealed about 270 differentially expressed proteins. Some of those proteins are related to bacterial interactions, response to antibiotic treatment or metabolic shifts. For instance, the enzymes PhzS(flavin-containing monooxygenase), PhzD (phenazine biosynthesis protein), and PhzG2 (pyridoxamine 5'-phosphate oxidase) indicated the production of the antibiotic pigment pyocyanine by P. aeruginosa that is related to oxidative stress and therefore, might inhibit growth of S. aureus. Overall, the strategy applied not only allows species-specific tracking of the community composition but also provides valuable insights into the behavior of mixed cultures.     

Metabolism of chlorinated methanes,ethanes, and ethylenes by a mixed bacterial culture growing on methane

Summary Soil was taken from the top 10 cm of a soil column that removed halogenated aliphatic hydrocarbons in the presence of natural gas. This soil was used as an enrichment inoculum to determine that the removals seen in the soil column were in fact of a microbiological nature. Methane served as the source of carbon and energy and was consumed immediately by the enrichments. After several transfers of the enrichments, a stable consortium of at least three bacterial types was obtained. The predominant bacterium was a non-motile, gram-negative coccus. This stable consortium was able to remove chlorinated methanes, ethanes, and ethylenes when grown with methane and oxygen in the headspace. Methane was required for the removals to be observed. Acetylene inhibited the removals, which further suggests the involvement of methanotrophs. Benzene and toluene were removed by the mixed culture with or without methane in the headspace. Fatty acid analysis of the mixed culture resulted in a profile that indicated that the predominant organism was a type II methanotroph. This study provides further evidence that methanotrophic bacteria are capable of cometabolizing a wide range of chlorinated methanes, ethanes, and ethylenes.     

Growth of a facultative anaerobe under oxygen-limiting conditions in pure culture and in co-culture with a sulfate-reducing bacterium

Abstract The occurrence and properties were studied of glucose-metabolizing bacteria present in the anaerobic sediment 5–10 cm below the surface of an estuarine tidal mud-flat. Of all these bacteria (10⁴– 10⁵ per g wet sediment) 80–90% were facultatively anaerobic species. Chemostat enrichments on glucose under aerobic, oxygen-limited and alternately aerobic-anaerobic conditions also yielded cultures dominated by facultative anaerobes. One of the dominant species, tentatively identified as a Vibrio sp., was studied in more detail under oxygen-limiting conditions. Fermentative and respiratory metabolisms were found to operate simultaneously, and the ratio between the two was regulated by the extent of oxygen limitation. A small fraction of the acetate formed under such growth conditions was shown to be subsequently respired. A co-culture was established of the Vibrio sp. and a sulfate-reducing bacterium ( Desulfovibrio HL21 ) in an aerated chemostat. The importance of these observations is discussed in relation to the role of facultative anaerobes in anaerobic habitats.     

Effects of media composition on substrate removal by pure and mixed bacterial cultures

Continuous culture experiments with identical experimental designs were run with a mixed microbial community of activated sludge origin and an axenic bacterial culture derived from it. Each culture received 2-chlorophenol (2-CP) at a concentration of 160 mg/L as COD and L-lysine at a concentration of 65 mg/L as COD. A factorial experimental design was employed with dilution rate and media composition as the two controlled variables. Three dilution rates were studied: 0.015, 0.0325, and 0.05 h^–1. Media composition was changed by adding four biogenic compounds (butyric acid, thymine, glutamic acid and lactose) in equal COD proportions at total concentrations of 0, 34, 225, and 1462 mg/L as COD. The measured variables were the effluent concentrations of 2-CP as measured by the 4-aminoantipyrene test and lysine as measured by the o-diacetylbenzene procedure. The results suggest that community structure and substrate composition play important roles in the response of a microbial community to mixed substrates. The addition of more biogenic substrates to the axenic culture had a deleterious effect on the removal of both lysine and 2-CP, although the effect was much larger on lysine removal. In contrast, additional substrates had a positive effect on the removal of 2-CP by the mixed community and much less of a negative effect on the removal of lysine. The dilution rate at which the cultures were growing had relatively little impact on the responses to the additional substrates.Abbreviations COD chemical oxygen demand - 2-CP 2-chlorophenol - DOC dissolved organic carbon - MDL method detection limit - SS suspended solids     

象山港网箱养殖对近海沉积物细菌群落的影响

微生物在有机物质分解、能量转移等生物地化循环中起着重要作用,研究养殖活动对沉积物中微生物多样性的影响对正确评价近海生态系统结构和功能具有重要意义.结合沉积物的理化分析(T、pH、Eh和SO2-),通过T-RFLP(末端限制性片段长度多态性)分析了象山港网箱养殖对近海不同深度沉积物中细菌群落结构和多样性的影响,构建了对照区0-5 cm沉积物样品的细菌16S rRNA基因克隆文库,并进行了系统发育分析.结果表明,近海网箱养殖活动不但使象山港沉积物Eh、pH值等理化性质有所改变,而且使细菌的群落结构也发生了变化,变形细菌纲细菌的相对丰度降低,细菌群落的多样性和均匀度也显著降低,致使近海沉积物细菌群落由多样性高的稳定结构向多样性降低的不稳定结构转变.     

转Bt基因玉米对根际土壤细菌群落结构的影响

利用聚合酶链式反应-变性梯度凝胶电泳(PCR-DGGE)技术及扩增产物序列分析方法研究了转Bt基因玉米对根际土壤细菌群落结构及系统发育的影响.结果表明:转基因玉米与其非转基因亲本根际共有的土壤细菌分别隶属于变形菌门、疣微菌门、放线菌门、拟杆菌门、厚壁菌门、芽单胞菌门和酸杆菌门.其中,变形菌门为主要优势类群,占43.5%...     

Cognitive optimization of microbial PHB production in an optimally dispersed bioreactor by single and mixed cultures

Cognitive (or intelligent) models are often superior to mechanistic models for nonideal bioreactors. Two kinds of cognitive models—cybernetic and neural—were applied recently to fed-batch fermentation by Ralstonia eutropha in a bioreactor with optimum finite dispersion. In the present work, these models have been applied in simulation studies of co-cultures of R. eutropha and Lactobacillus delbrueckii. The results for both cognitive and mechanistic models have been compared with single cultures. Neural models were the most effective for both types of cultures and mechanistic models the least effective. Simulations with co-culture fermentations predicted more PHB than single cultures with all three types of models. Significantly, the predicted enhancements in PHB concentration by cognitive methods for mixed cultures were four to five times larger than the corresponding increases in biomass concentration. Further improvements are possible through a hybrid combination of all three types of models.     

Improvement of polygalacturonase production at high temperature by mixed culture of Aspergillus niger and Saccharomyces cerevisiae

Catabolic repression in the synthesis of inducible enzymes by glucose, fructose, and intermediates of the glycolytic cycle has been observed in many microorganisms. In order to enhance the polygalacturonase (PG) production of Aspergillus niger GJ-2, Saccharomyces cerevisiae J-1 was inoculated to the medium at 12 h of culture, which resulted in a significant improvement of PG production. It was also found that maximum PG activity of 512.7 U/ml was obtained at 37 °C in the mixed culture, which was nearly twofold higher than that of the culture without the inoculation of S. cerevisiae J-1.     

象山港网箱养殖区水体和沉积物的细菌生态分布

2006年1月和10月对象山港网箱养殖区细菌数量分布及群落结构进行研究,并与周边非养殖海域进行了比较。结果显示水体和沉积物细菌数量均呈现10月份1月份、网箱养殖区网箱外侧对照区近湾顶对照区近湾口对照区的分布特点,其中网箱养殖区水体异养细菌、弧菌数量分别在1.0×106—7.2×107CFU/L和1.6×105—1.3×106CFU/L之间,沉积物异养细菌、弧菌数量分别在5.9×102—6.1×105CFU/g和8.0×101—7.4×104CFU/g之间。网箱养殖区水体分离细菌89株,归属于12个属,其中不动杆菌属(Acinetobacter)、假单胞菌属(Pseudomonas)、弧菌属(Vibrio)等为优势菌属;沉积物分离细菌136株,归属于10个属,其中假单胞菌属(Pseudomonas)、弧菌属(Vibrio)、芽孢杆菌属(Bacillus)、不动杆菌属(Acinetobacter)等为优势菌属。由于长期受到高营养盐、高有机质等外界条件的选择作用,网箱区及网箱外侧对照区海域的细菌群落多样性较低,某些特定的细菌类群富集。研究表明网箱养殖区的细菌分布和菌属组成特征与养殖自身污染密切相关,并且养殖过程对细菌群落产生的影响已扩展至网箱外侧的邻近海域。     

低剂量混合稀土积累对黄褐土微生物主要类群的生态效应

采用田间小区试验和室内低剂量模拟叠加试验相结合的方法,研究低剂量混合稀土在黄褐土中积累对土壤微生物主要类群的生态效应．结果表明,低剂量稀土的持续积累对土壤细菌、放线菌产生刺激、抑制、再刺激的交替作用;对真菌也产生类似的作用,但抑制作用不显著,而刺激作用持续、明显．混合稀土对3类土壤微生物数量抑制程度顺序为：细菌>放线菌>霉菌．稀土积累至150mg·kg^-1时,土壤各类微生物的种群结构均发生显著的改变,耐稀土微生物数量大幅度增加,细菌中的G^-细菌、链霉菌的白孢类群、真菌中青霉分别成为优势种群．对低浓度稀土积累的田问土壤微生物学参数模拟计算结果表明,稀土对土壤细菌、放线菌和真菌的EC50(半抑制浓度)值分别为24．1、41．6～73．8和55．3～150．1mg·kg^-1,30mg·kg^-1值可以初步确定为稀土在黄褐土中积累的安全临界值．     

Clumped and isolated trees influence soil nutrient levels in an Australian temperate box woodland

Isolated paddock trees are a common feature of agri-pastoral landscapes in south-eastern Australia. We assessed the impact of trees on soil nutrients by examining (1) changes in soil nutrients under clumped and isolated (living and dead) trees at four microsites corresponding with increasing distance from the trunk (trunk, mid-canopy, drip line, open), and (2) changes with depth under trees growing in clearly-defined clumps. We detected significantly greater concentrations of organic C, and total N and S under trees growing in clumps compared with either isolated living or dead trees. Levels of soluble Ca²⁺, K⁺ and Mg²⁺, pH, electrical conductivity (EC) and available P declined with increasing distance from the trees, but there were no significant trends for organic C, or total N and S. The concentration of most nutrients declined with depth, particularly at microsites close to the trunk, while pH increased with depth. We believe that differences in chemistry were largely driven by greater inputs of organic matter under the trees. This study reinforces the view that trees, whether scattered or in clumps, are important for soil nutrient conservation in agri-pastoral landscapes.     

Effects of sulfate on lactate and C2-, C3- volatile fatty acid anaerobic degradation by a mixed microbial culture

The effects of sulfate on the anaerobic degradation of lactate, propionate, and acetate by a mixed bacterial culture from an anaerobic fermenter fed with wine distillery waste water were investigated. Without sulfate and with both sulfate and molybdate, lactate was rapidly consumed, and propionate and acetate were produced; whereas with sulfate alone, only acetate accumulated. Propionate oxidation was strongly accelerated by the presence of sulfate, but sulfate had no effect on acetate consumption even when methanogenesis was inhibited by chloroform. The methane production was not affected by the presence of sulfate. Counts of lactate- and propionate-oxidizing sulfate-reducing bacteria in the mixed culture gave 4.5×10⁸ and 1.5×10⁶ viable cells per ml, respectively. The number of lactate-oxidizing fermentative bacteria was 2.2×10⁷ viable cells per ml, showing that sulfate-reducing bacteria outcompete fermentative bacteria for lactate in the ecosystem studied. The number of acetoclastic methanogens was 3.5×10⁸ viable cells per ml, but only 2.5×10⁴ sulfate reducers were counted on acetate, showing that acetotrophic methanogens completely predominated over acetate-oxidizing sulfate-reducing bacteria. The contribution of acetate as electron donor for sulfate reduction in the ecosystem studied was found to be minor.     

茶园根际土壤细菌群落结构及多样性

【背景】茶园根际土壤的细菌群落结构与茶园生境土壤营养循环密切相关,其组成及多样性可以作为健康茶园的一个生物指标。【方法】采用PCR．变性梯度凝胶电泳（PCR—DGGE）分子指纹图谱技术,检测安溪铁观音种植区不同海拔茶园根际土壤样本的细菌群落结构,利用Shannon．Wiener多样性指数分析其多样性,采用非加权组平均法进行聚类分析得到其分布特征,利用蒙特卡罗检验和冗余分析分别揭示影响细菌群落分布的环境因子及细菌群落分布和环境变量之间的关系。【结果】茶园根际土壤细菌的DGGE结果显示,检测到的14种主要细菌中有11种细菌是不可培养的,3种细菌是可培养的,分别属于根瘤菌属、中华根瘤菌属和苍白杆菌属。聚类分析得到,同一海拔梯度茶园根际土壤细菌群落结构相似。Shan-non—Wiener多样性指数分析表明,400m海拔处茶园根际土壤细菌群落多样性最高。蒙特卡罗检验分析得到环境因子协同作用对茶园根际土壤细菌群落结构贡献率为59．6％。冗余分析显示,茶园根际土壤细菌群落结构与海拔密切相关。【结论与意义】茶园根际土壤细菌群落结构分布与海拔梯度密切相关,考虑不同海拔高度土壤细菌群落对茶园营养循环的影响,在铁观音的健康栽培和管理过程中具有重要意义。