Cadmium Interaction with Microalgal Cells,Cyanobacterial Cells,and Seaweeds; Toxicology and Biotechnological Potential for Wastewater Treatment

The accumulation of cadmium (Cd) by Tetraselmis chuii and Spirulina maxima was studied with dead and growing cells. Results indicated that the 2 microorganisms accumulated Cd by 2 different means according to the mechanisms involved—metabolism-dependent or metabolism-independent sorption. The mechanism involved in Cd accumulation on Tetraselmis chuii was restricted to surface phenomena, while in Spirulina maxima, Cd was accumulated on different layers of the cyanobacterium surface. In order to select a suitable immobilization support for the cells, several seaweeds were tested. Two types of seaweed were selected for experiments, using a small continuous pilot unit: Sargassum sp., a strong Cd adsorber, and Ulva sp., a poor one. The column reactors of the continuous system were filled with the algal supports and covered with dense microbial biofilms of Tetraselmis chuii or Spirulina maxima. The results obtained proved the success of the association between living microbial cells and dead seaweeds for operation of the continuous system.     

合成生物学方法构建电活性大肠杆菌

电活性微生物具有独特的在细胞内外环境之间传递电子的能力。在对天然电活性微生物电子传递机制充分研究的基础上,通过合成生物学方法异源构建天然电活性微生物电子传递结构基础也可以将遗传背景清晰的非电活性大肠杆菌改造为电活性微生物。构建获得的工程化电活性大肠杆菌可以直接应用于微生物燃料电池和生物传感器等领域,同时也可以作为底盘细胞整合相应的目标产物合成通路实现电能驱动的生物合成。本文以合成生物学方法构建电活性大肠杆菌为主题,详细阐述天然电活性微生物电子传递的机理及结构基础,总结了工程化电活性大肠杆菌的构建策略、成功案例以及应用领域,并对合成生物学方法构建电活性大肠杆菌未来的研究方向进行了展望。     

Uptake of volatilen-alkanes byPseudomonas PG-I

Using a bacterial speciesPseudomonas PG-1, evidence has been obtained which indicates that uptake ofn-pentane ton-octane by microbial cells takes place primarily from the gas phase either directly orvia the aqueous phase. Specific growth rate increased along with the increase in substrate concentration but above the alkane concentration of 0.3% by volume, specific growth rate decreased indicating substrate inhibition of growth. In the case of less volatile alkanes,n-nonane andn-decane, substrate transfer is predominantly through substrate solubilization system elaborated by the cells. EDTA, a strong inhibitor of hydrocarbon solubilization by the cells, inhibited growth on these two alkanes but had negligible effect on growth onn-pentane ton-octane.     

Lactic acid fermentation by cells of Lactobacillus rhamnosus immobilized in polyacrylamide gel

Summary The process of lactic acid fermentation of lactose to lactic acid by Lactobacillus rhamnosus ATCC 7469 has been studied. The following processes have been explored: growth kinetics, as well as lactose utilization, production of lactic acid and further degradation of lactic acid. The immobilization experiments were conducted with microbial cells entrapped in polyacrylamide gels. Gels with different ratios of the monomer (acrylamide) and the cross-linking agent (N,N′-methylene-bis-acrylamide) have been tested. These were used in a repeat-batch process. The current processes inside and outside the gel particles were subjects of examination. The evolution of the activity of immobilized cells with repeated use showed that the particles served mainly as a donor of cells for the free culture. In all experiments a very high degree of conversion, 85–90% was observed. After several runs however, the particles were exhausted for microbial cells. A kinetic model of the process of lactic acid production was developed. This model allowed the evaluation of the effect of microbial growth and diffusion limitations inside the gel particles on the process rate and the separate contribution of the free and immobilized cells to the overall fermentation process upon multiple use.     

Starter culture of <Emphasis Type="Italic">Pseudomonas veronii</Emphasis> strain B for aerobic granulation

Aggregation of bacterial cells is used in formation of microbial granules. Aerobically grown microbial granules can be used as the bio-agents in the treatment of wastewater. However, there are problems with start up of microbial granulation and biosafety of this process. Aim of this research was selection and testing of safe microbial strain with high cell aggregation ability to shorten period of microbial granules formation. Five bacterial strains with cell aggregation index higher than 50% have been isolated from the granules. Strain of Pseudomonas veronii species was considered as most probably safe starter culture for granulation because other strains belonged to the species known as human pathogens. The microbial granules were formed after 3 days of cultivation in case when P. veronii strain B was applied to start-up aerobic granulation process using model wastewater. The granules were produced from activated sludge after 9 days of cultivation. Microbial aggregates produced from starter culture of P. veronii strain B were more compact (sludge volume index was 70 ml/g) than those produced from activated sludge (sludge volume index was 106 ml/g). It is a first proof that application of selected safe starter pure culture with high cell aggregation ability can accelerate and enhance formation of microbial granules.     

香坊肠球菌和罗伊氏乳杆菌在无菌猪肠道中的定殖

【背景】已有研究表明,微生物在宿主肠道中的定殖受宿主、肠道环境、微生物物种特性和菌株来源等多个因素的影响。一般认为,来源于同类宿主的微生物菌株,在该类宿主肠道中具有定殖优势,但缺乏在物种和菌株水平上研究微生物自身特性在宿主肠道中定殖的研究报道。【目的】将不同来源(同类宿主肠道、非同类宿主肠道和非肠道环境)、具有不同生物学特性的3株香坊肠球菌(Enterococcus xiangfangensis)和4株罗伊氏乳杆菌(Lactobacillusreuteri)对无菌猪肠道进行定殖,在物种和菌株2个水平上探究物种特性和菌株来源对宿主肠道定殖的偏好性,揭示影响微生物定殖效率的关键因素。【方法】在本项研究中,将从藏猪(Tibetan pigs)、小鼠(ob/ob mice)、食蟹猴(Macaca fascicularis)和发酵食品中分离得到的多株香坊肠球菌和罗伊氏乳杆菌,制成混合菌剂对无菌巴马香猪(Bama miniature pig)进行为期4周的饲喂,并通过实时荧光定量PCR方法检测这7株菌在无菌猪肠道中的定殖情况。【结果】在物种水平上,香坊肠球菌和罗伊氏乳杆菌在无菌猪体内具有相近的定殖...     

生物量浓度实时在线检测方法的研究

微生物的存在会改变发酵液的电特性,发酵液在无线电频率范围内的电容率增量是测量频率和生物量浓度的函数.基于对发酵液电容率分布的研究,提出了测量生物量浓度的新方法.用此方法不用取样就能对发酵液中的生物量进行实时在线测量,而且测得的是活的生物量浓度.制作的电极直接插入发酵器中并满足高温蒸气灭菌条件.此方法在生化制药、食品发酵、啤酒酿造、污水检测等工业领域里有很好的推广应用前景.     

Conversion of<Emphasis Type="Italic">G. hansenii</Emphasis> PJK into non-cellulose-producing mutants according to the culture condition

The conversion of a cellulose-producing cell (Cel ⁺) fromGluconacetobacter hansenii PJK (KCTC 10505 BP) to a non-cellulose-producing cell (Cel ⁻) was investigated by measuring the colony forming unit (CFU). This was achieved in a shaking flask with three slanted baffles, which exerted a strong shear stress. The addition of organic acid, such as glutamic acid and acetic acid, induced the conversion of microbial cells from a wild type toCel ⁻ mutants in a flask culture. The supplementation of 1% ethanol to the medium containing an organic acid depressed the conversion of the microbial cells toCel ⁻ mutants in a conventional flask without slanted baffles. The addition of ethanol to the medium containing an organic acid; however, accelerated the conversion of microbial cells in the flask with slanted baffles. TheCel ⁺ cells from the agitated culture were not easily converted intoCel ⁻, mutants on the additions of organic acid and ethanol to a flask without slanted baffles, but some portion of theCel ⁺ cells were converted toCel ⁻ mutants in a flask with slanted baffles. The conversion ratio ofCel ⁺ cells toCel ⁻ mutants was strongly related to the production of bacterial cellulose independently from the cell growth.     

The role of released ATP in killing Candida albicans and other extracellular microbial pathogens by cationic peptides

A unifying theme common to the action of many cationic peptides that display lethal activities against microbial pathogens is their specific action at microbial membranes that results in selective loss of ions and small nucleotides—chiefly ATP. One model cationic peptide that induces non-lytic release of ATP from the fungal pathogen Candida albicans is salivary histatin 5 (Hst 5). The major characteristic of Hst 5-induced ATP release is that it occurs rapidly while cells are still metabolically active and have polarized membranes, thus precluding cell lysis as the means of release of ATP. Other cationic peptides that induce selective release of ATP from target microbes are lactoferricin, human neutrophil defensins, bactenecin, and cathelicidin peptides. The role of released extracellular ATP induced by cationic peptides is not known, but localized increases in extracellular ATP concentration may serve to potentiate cell killing, facilitate further peptide uptake, or function as an additional signal to activate the host innate immune system at the site of infection.     

Thermus 2S from Thai hot springs: isolation and immobilization

Three strains of thermophilic bacteria producing extracellular protease have been isolated from hot springs in Chiang Mai, Thailand. The bacterium producing the highest amount of protease has been selected and identified as belonging to the genusThermus, and is tentatively calledThermus 2S. The isolate is a Gram-negative, rod shaped bacterium. It exhibited maximum growth around 60°C at pH 7. Entrapment of the microbial cells in calcium alginate maintained the cell viability. Protease production from immobilized cells using 2 g wet cells per 10 ml 3% (w/v) sodium alginate was higher than that from a free-cell system using 2% inoculum.     

The influence of cultivation methods on Shewanella oneidensis physiology and proteome expression

High-throughput analyses that are central to microbial systems biology and ecophysiology research benefit from highly homogeneous and physiologically well-defined cell cultures. While attention has focused on the technical variation associated with high-throughput technologies, biological variation introduced as a function of cell cultivation methods has been largely overlooked. This study evaluated the impact of cultivation methods, controlled batch or continuous culture in bioreactors versus shake flasks, on the reproducibility of global proteome measurements in Shewanella oneidensis MR-1. Variability in dissolved oxygen concentration and consumption rate, metabolite profiles, and proteome was greater in shake flask than controlled batch or chemostat cultures. Proteins indicative of suboxic and anaerobic growth (e.g., fumarate reductase and decaheme c-type cytochromes) were more abundant in cells from shake flasks compared to bioreactor cultures, a finding consistent with data demonstrating that “aerobic” flask cultures were O₂ deficient due to poor mass transfer kinetics. The work described herein establishes the necessity of controlled cultivation for ensuring highly reproducible and homogenous microbial cultures. By decreasing cell to cell variability, higher quality samples will allow for the interpretive accuracy necessary for drawing conclusions relevant to microbial systems biology research.     

Quantification of ratios of Bacteria and Archaea in methanogenic microbial community by fluorescence in situ hybridization and fluorescence spectrometry

16S rRNA-targeted oligonucleotide probes for Bacteria (Eub338) and Archaea (Arc915) were used for whole-cell, fluorescence in situ hybridization (FISH) to quantify the ratio of these microbial groups in an anaerobic digester. The quantity of specifically bound (hybridized) probe was measured by fluorescence spectrometry and evaluated by analysing the dissociation curve of the hybrids, by the measurement of the binding with a nonsense probe, and by the competitive inhibition of the binding of the labelled probe by the corresponding unlabelled probe. Specific binding of oligonucleotide probes with the biomass of anaerobes was 40–50% of their total binding. The ratio of Arc915 and Eub338 probes hybridized with rRNAs of the cells in anaerobic sludge was 0.50. Measurement of FISH by fluorescence spectrometry appears to be a suitable method for quantification of the microbial community of anaerobes.     

The Effect of the Intra- and Extracellular Metabolites of Microorganisms Isolated from Various Ecotopes on the Catalase Activity of Staphylococcus aureus ATCC 6538 P

The cell extracts (i.e., intracellular metabolites) and culture liquids (i.e., extracellular metabolites) of microorganisms isolated from various ecotopes were found to inhibit the catalase activity of Staphylococcus aureus ATCC 6538 P, which resulted in a considerable inhibition of the growth of metabolite-treated S. aureus cells by hydrogen peroxide. The inhibitory effect of microbial metabolites on S. aureus catalase can be considered as a mechanism of intercellular interactions responsible for the formation of microbiocenoses.     

Histological changes associated with acquisition of competence for shoot regeneration in leaf discs ofSaintpaufla ionantha xconfusa hybrid (African violet) cultured in vitro

In leaf discs ofSaintpaulia ionantha xconfusa hybrid (cv. Virginia) cultured on shoot-inducing medium, periclinal divisions were initiated in epidermal cells 3–5 days after explant isolation. This timing coincided with the time for competence acquisition determined in tissue-transfer experiments. Some of the daughter cells from periclinal divisions formed the target cells which divided both anticlinally and periclinally to form cell division centers (meristemoids), precursors of adventitious shoots. The target cells were not morphologically distinct from other epidermal cells at the light microscope level. It is suggested that the periclinal divisions in epidermal cells represent the dedifferentiation phase during which target (competent) cells are formed. Once the cells have acquired the ability to divide periclinally, both dedifferentiation and shoot induction occur in the presence of exogenous plant hormones.Abbreviations SIM Shoot-inducing medium     

降解1,3-二甲基-2-咪唑烷酮细菌的分离及功能评价

[背景] 1,3-二甲基-2-咪唑烷酮（1,3-Dimethyl-2-Imidazolidinone,DMI）作为一种强极性非质子溶剂,在生产和应用过程的环境中有稳定残留问题,存在安全隐患。[目的] 分离筛选具有降解DMI能力的微生物菌株,为清除环境中残留的DMI提供优良的微生物菌种资源。[方法] 从DMI生产区域土壤采集样品分离DMI抗性微生物,采用形态学及分子生物学鉴定确定其分类地位,并对DMI降解能力进行测定。[结果] 分离到最高能够耐受5%（体积分数） DMI的微生物菌株,形态学及分子生物学鉴定初步表明获得的菌株DT-1和DT-2为贝莱斯芽孢杆菌（Bacillus velezensis）;全细胞及细胞提取液均具有降解DMI的能力;其中菌株DT-1及其细胞提取液对1%（体积分数） DMI的降解率分别达到48%和68%。[结论] 从DMI生产区域土壤中分离到具有DMI降解能力的芽孢杆菌,不但可为DMI污染的微生物治理提供优良微生物资源,而且扩展了人们对芽孢杆菌生物学功能的认识。     

The trade-off between growth rate and yield in microbial communities and the consequences for under-snow soil respiration in a high elevation coniferous forest

Soil microbial respiration is a critical component of the global carbon cycle, but it is uncertain how properties of microbes affect this process. Previous studies have noted a thermodynamic trade-off between the rate and efficiency of growth in heterotrophic organisms. Growth rate and yield determine the biomass-specific respiration rate of growing microbial populations, but these traits have not previously been used to scale from microbial communities to ecosystems. Here we report seasonal variation in microbial growth kinetics and temperature responses (Q₁₀) in a coniferous forest soil, relate these properties to cultured and uncultured soil microbes, and model the effects of shifting growth kinetics on soil heterotrophic respiration (R_h). Soil microbial communities from under-snow had higher growth rates and lower growth yields than the summer and fall communities from exposed soils, causing higher biomass-specific respiration rates. Growth rate and yield were strongly negatively correlated. Based on experiments using specific growth inhibitors, bacteria had higher growth rates and lower yields than fungi, overall, suggesting a more important role for bacteria in determining R_h. The dominant bacteria from laboratory-incubated soil differed seasonally: faster-growing, cold-adapted Janthinobacterium species dominated in winter and slower-growing, mesophilic Burkholderia and Variovorax species dominated in summer. Modeled R_h was sensitive to microbial kinetics and Q₁₀: a sixfold lower annual R_h resulted from using kinetic parameters from summer versus winter communities. Under the most realistic scenario using seasonally changing communities, the model estimated R_h at 22.67 mol m⁻² year⁻¹, or 47.0% of annual total ecosystem respiration (R_e) for this forest.     

Evaluation of the growth of microorganisms on diaper absorbent materials

Summary Methods were developed to study the effects of absorbent materials from diapers on microbial survival, growth and toxic shock syndrome toxin-1 (TSST-1) production under specified in vitro conditions. Growth of representative skin and fecal flora organisms was equivalent in cultures in which materials from cotton cloth diapers, disposable diapers or disposable diapers containing absorbent gelling material were added as the sole carbon source. In urine used as an enrichment medium, growth of the test organisms in media containing material from the three diaper types was equivalent and no contribution to growth from the diaper material was detected. TSST-1 was not produced byStaphylococcus aureus under conditions in which urine was added to the diaper materials. Pathogenic strains of organisms purposefully introduced onto diapers failed to survive and the few microbial cells normally found in diaper material did not multiply when stored under conditions favorable to microbial growth. The data indicate that all three diaper types tested were the same with respect to growth and survival of representative skin and fecal organisms.     

An examination of the role of colonial <Emphasis Type="Italic">Phaeocystis antarctica</Emphasis> in the microbial food web of the Ross Sea

The extensive buildup of phytoplankton biomass in the Ross Sea conflicts with the view that high rates of herbivory occur in all regions of the Southern Ocean. Nano and microplanktonic consumers comprise a significant fraction of total plankton biomass; however, the importance of grazing remains uncertain in the Ross Sea. Microzooplankton ingestion of solitary and colonial cells of Phaeocystis antarctica were calculated using a novel live-staining fluorescently-labeled algae method. Different morphotypes of P. antarctica were stained different colors, mixed, and observed inside Euplotes to determine their feeding preference. The blue (7-aminocoumarin) (CMAC) stain was used on the colonies and the green (CMFDA) CellTracker Probe was used on solitary cells. Both morphotypes can be seen inside the food vacuoles of the ciliate, supporting the idea that microzooplankton are capable of ingesting cells within the colonial matrix. This suggests that P. antarctica colonies enter the microbial loop in the Ross Sea before sedimentation.     

Production of l (+) lactic acid by Lactobacillus delbrueckii immobilized in functionalized alginate matrices

The role of functionalized alginate gels as immobilized matrices in production of l (+) lactic acid by Lactobacillus delbrueckii was studied. L. delbrueckii cells immobilized in functionalized alginate beads showed enhanced bead stability and selectivity towards production of optically pure l (+) lactic acid in higher yields (1.74Yp/s) compared to natural alginate. Palmitoylated alginate beads revealed 99% enantiomeric selectivity (ee) in production of l (+) lactic acid. Metabolite analysis during fermentation indicated low by-product (acetic acid, propionic acid and ethanol) formation on repeated batch fermentation with functionalized immobilized microbial cells. The scanning electron microscopic studies showed dense entrapped microbial cell biomass in modified immobilized beads compared to native alginate. Thus the methodology has great importance in large-scale production of optically pure lactic acid.