Kinetics of biodegradation of p-xylene and naphthalene and oxygen transfer in a novel airlift immobilized bioreactor

The scope of this study included the biodegradation performance and the rate of oxygen transfer in a pilot-scale immobilized soil bioreactor system (ISBR) of 10-L working volume. The ISBR was inoculated with an acclimatized population of contaminant degrading microorganisms. Immobilization of microorganisms on a non-woven polyester textile developed the active biofilm, thereby obtaining biodegradation rates of 81 mg/L x h and 40 mg/L x h for p-xylene and naphthalene, respectively. Monod kinetic model was found to be suitable to correlate the experimental data obtained during the course of batch and continuous operations. Oxygen uptake and transfer rates were determined during the batch biodegradation process. The dynamic gassing-out method was used to determine the oxygen uptake rate (OUR) and volumetric oxygen mass transfer, K(L) a. The maximum volumetric OUR of 255 mg O(2)/L x h occurred approximately at 720-722 h after inoculation, when the dry weight of biomass concentration was 0.67 g/L.     

Continuous, real-time monitoring of the oxygen uptake rate (OUR) in animal cell bioreactors

A new method for real-time monitoring of the oxygen uptake rate (OUR) in bioreactors, based on dissolved oxygen (DO) measurement at two points, has been developed and tested extensively. The method has several distinct advantages over known techniques.It enables the continuous and undisturbed monitoring of OUR, which is conventionally impossible without gas analyzers. The technique does not require knowledge of k(L)a. It provides smooth, robust, and reliable signal. The monitoring scheme is applicable to both microbial and mammalian cell bioprocesses of laboratory or industrial scale. The method was successfully used in the cultivation of NSO-derived murine myeloma cell line producing monoclonal antibody. It was found that while the OUR increased with the cell density, the specific OUR decreased to approximately one-half at cell concentrations of 16 x 10(6) cells/mL, indicating gradual reduction of cell respiration activity. Apart from the laboratory scale cultivation, the method was applied to industrial scale perfusion culture, as well as to processes using other cell lines. (c) 1994 John Wiley & Sons, Inc.     

应用动力学方法在线检测Vero细胞培养过程中的摄氧率

流加和灌注培养已被广泛应用于动物细胞培养 ,以获得高活性、高密度的细胞和高的产物得率。在这些培养过程中 ,一般通过离线检测关键参数 (如细胞密度、营养和代谢产物的浓度 )来人为调整灌注速率和补料策略 ,但是 ,当细胞密度较高时 ,由于细胞代谢旺盛使得培养的微环境变化很快 ,这就需要更加频繁快速地调整操作条件 ,从而导致因频繁取样和离线分析所带来的污染危险及大量人力、物力的浪费。这在大规模细胞培养过程中是不可取的。因此 ,要建立大规模、高效动物细胞培养过程 ,有必要研究和探索在线检测技术 ,以实时掌握细胞培养过程所处的状…     

Oxygen transfer enhancement in aqueous/perfluorocarbon fermentation systems: I. experimental observations

A new fiber-optic dissolved oxygen sensing technique was applied to the study of two-phase aqueous/perfluorocarbon (pfc) dispersions. These dispersions were examined for their oxygen transfer enhancement capability in the absence and presence of an oxygen-consuming reaction. For the pfc-in-water dispersions, oxygen uptake rate (OUR) enhancements were equal both with and without oxygen-consuming cells present in the aqueous phase. In contrast, for water-in-pfc dispersions, OUR enhancements inthe presence of reaction were limited by oxygen diffusion across the aqueous phase droplets. Nevertheless, enhancement factors of 5-10 on an aqueous phase volume basis were obtained in a 75% pfc dispersion.These oxygen transfer enhancements were directly translatable into enhancements in overall fermenter productivity for actual microbial cultivation systems.     

现代分子生物学技术在瘤胃微生态系统研究中的应用

瘤胃中栖息着大量的微生物,由于这些微生物组成复杂且有些细菌在体外无法培养,目前对这些微生物的了解仍然很少。现代分子生物学技术的发展为研究瘤胃微生物提供了有效的方法,利用核酸探针、基因序列分析、遗传指纹技术、全细胞杂交和实时定量PCR等技术可以对瘤胃微生物的分类及进化关系、区系结构图、重要酶的表达以及目的微生物的准确定量进行更为深入和透彻的研究。发展和利用这些技术不仅可以研究微生物之间的关系以及微生物与饲料颗粒之间时间与空间的关系,还能直接在细菌自然生长的环境中对其各种特征进行研究。     

稳定性同位素探测技术在微生物生态学研究中的应用

稳定性同位素标记技术同分子生物学技术相结合而发展起来的稳定性同位素探测技术（stableisotope probing,SIP）,在对各种环境中微生物群落组成进行遗传分类学鉴定的同时,可确定其在环境过程中的功能,提供复杂群落中微生物相互作用及其代谢功能的大量信息,具有广阔的应用前景.其基本原理是：将原位或微宇宙（microcosm）的环境样品暴露于稳定性同位素富集的基质中,这些样品中存在的某些微生物能够以基质中的稳定（性同位素为碳源或氮源进行物质代谢并满足其自身生长需要,基质中的稳定性同位素被吸收同化进入微生物体内,参与各类物质如核酸（DNA和RNA）及磷脂脂肪酸（PLFA）等的生物合成,通过提取、分离、纯化、分析这些微生物体内稳定性同位素标记的生物标志物,从而将微生物的组成与其功能联系起来.在介绍稳定性同位素培养基质的选择及标记方法、合适的生物标志物的选择及提取分离方法的基础上,举例阐述了此项技术在甲基营养菌、有机污染物降解菌、根际微生物生态、互营微生物、宏基因组学等方面的应用.     

熏蒸剂溴甲烷对农田土壤微生物的影响

土壤微生物是反应土壤健康状况最敏感的生物学指标,溴甲烷残留不仅消耗臭氧层,影响生态平衡,还会造成土壤质量恶化和微生物群落结构的变化。为明确溴甲烷对农田土壤微生物群落结构及生态过程的影响,以兰州市红古区连续两年种植草莓的土壤为研究对象,测定熏蒸剂溴甲烷处理后土壤微生物量碳、基础呼吸、诱导呼吸和微生物代谢熵等相关指标,并运用磷脂脂肪酸法(PLFA)测定不同类群微生物的变化。结果表明:经溴甲烷熏蒸处理至培养结束(第90天)时土壤微生物基础呼吸和诱导呼吸分别下降0.6%和16.2%,并且与对照皆差异显著(P 0.05);微生物量碳培养结束时与对照差异显著(P 0.05),且减少5.6%;总体上微生物基础呼吸、诱导呼吸和微生物量碳都呈现先下降后逐渐恢复的趋势;微生物代谢熵(q CO_2)第15天后都高于对照,但随培养时间延长,处理组和对照组的差值逐渐降低,到培养期结束仍未恢复,相差5.1%。溴甲烷对土壤细菌(B)、真菌(F)和革兰氏阴性菌(GN)、革兰氏阳性菌(GP)都存在抑制作用; B、F含量分别较对照下降0.64%—8.72%、0.03%—5.61%;到培养期结束时,GP的量下降0.26%,GN下降10.42%,GN对溴甲烷的敏感性强于GP,且GN的变化具有滞后性;溴甲烷处理降低了B/F和GN/GP,但对GN/GP影响比对B/F的更为显著,土壤微生物压力指数增加。综上,说明施用溴甲烷使农田土壤微生物受到了长期的、持续的外源压力胁迫,溴甲烷在对有害微生物杀死的同时,也对有益微生物造成极大的伤害,不利于土壤优良性状的保持,使土壤中微生物丰富度和多样性下降。因此,实际应用中应充分考虑溴甲烷对土壤微生物带来的负面影响。     

Short-term effects of rhizosphere microorganisms on fe uptake from microbial siderophores by maize and oat

Effects of rhizosphere microorganisms on Fe uptake by oat (Avena sativa) and maize (Zea mays) were studied in short-term (10 h) nutrient solution experiments. Fe was supplied either as microbial siderophores (pseudobactin [PSB] or ferrioxamine B [FOB]) or as phytosiderophores obtained as root exudates from barley (epi-3-hydroxy-mugineic acid [HMA]) under varied population densities of rhizosphere microorganisms (axenic, uninoculated, or inoculated with different microorganism cultures). When maize was grown under axenic conditions and supplied with FeHMA, Fe uptake rates were 100 to 300 times higher compared to those in plants supplied with Fe siderophores. Fe from both sources was taken up without the involvement of an extracellular reduction process. The supply of FeHMA enhanced both uptake rate and translocation rate to the shoot (more than 60% of the total uptake). However, increased density of microorganisms resulted in a decrease in Fe uptake rate (up to 65%), presumably due to microbial degradation of the FeHMA. In contrast, when FeFOB or FePSB was used as the Fe source, increased population density of microorganisms enhanced Fe uptake. The enhancement of Fe uptake resulted from the uptake of FeFOB and FePSB by microorganisms adhering to the rhizoplane or living in the free space of cortical cells. The microbial apoplastic Fe pool was not available for root to shoot transport or, thus, for utilization by the plants. These results, in addition to the low uptake rate under axenic conditions, are in contrast to earlier hypotheses suggesting the existence of a specific uptake system for Fe siderophores in higher plants. The bacterial siderophores PSB and FOB were inefficient as Fe sources for plants even when supplied by stem injection. It was concluded that microorganisms are involved in degradation processes of microbial siderophores, as well as in competition for Fe with higher plants.     

油藏微生物群落研究的方法学

油藏微生物群落的解析和认知是开发和应用微生物采油技术的基础。利用各种提高油藏微生物可培养性的方法和非培养技术解析不同油藏微生物的群落结构、功能和多样性,对定向调控油藏微生物群落、开发和应用有效微生物驱油技术具有重要的指导意义。通过调查新近发展的提高微生物可培养性的方法和措施以及不依赖于培养的分子微生物生态学技术,总结了油藏微生物群落研究方法学的最新进展。提高微生物可培养性的方法和措施主要通过模拟微生物的生存环境,减少富营养的毒害作用、添加信号分子维持微生物细胞间的作用和提供新型电子供体和受体等手段采用稀释法、高通量培养法等方法得以实现;不依赖于培养的分子微生物生态学技术主要包括荧光原位杂交、末端限制性片断长度多态性分析、变性梯度凝胶电泳和构建克隆文库等技术。这些方法学的进展为更有效的获得各种油藏微生物资源、调控油藏微生物群落以提高石油采收率提供理论指导。     

Impact of an indigenous microbial enhanced oil recovery field trial on microbial community structure in a high pour-point oil reservoir

Based on preliminary investigation of microbial populations in a high pour-point oil reservoir, an indigenous microbial enhanced oil recovery (MEOR) field trial was carried out. The purpose of the study is to reveal the impact of the indigenous MEOR process on microbial community structure in the oil reservoir using 16Sr DNA clone library technique. The detailed monitoring results showed significant response of microbial communities during the field trial and large discrepancies of stimulated microorganisms in the laboratory and in the natural oil reservoir. More specifically, after nutrients injection, the original dominant populations of Petrobacter and Alishewanella in the production wells almost disappeared. The expected desirable population of Pseudomonas aeruginosa, determined by enrichment experiments in laboratory, was stimulated successfully in two wells of the five monitored wells. Unexpectedly, another potential population of Pseudomonas pseudoalcaligenes which were not detected in the enrichment culture in laboratory was stimulated in the other three monitored production wells. In this study, monitoring of microbial community displayed a comprehensive alteration of microbial populations during the field trial to remedy the deficiency of culture-dependent monitoring methods. The results would help to develop and apply more MEOR processes.     

Predicting the effects of chlorine on the micro-organisms of filamentous bulking activated sludges

Rapid and definite assessment of the effect that a specific biocide has on a specific case of filamentous bulking sludge is a much-needed tool in activated sludge wastewater treatment. The Live/Dead stain (LIVE/DEAD BacLight) distinguishing "living" and "non-living" cells, a nitrifying activity (NA) test and the oxygen uptake rate (OUR) measurement were examined for their appropriateness to predict the effects of chlorine on filamentous bulking sludges. The study showed the live/dead stain to be relevant for revealing the specific effect of chlorine on the filamentous bacteria of a bulking sludge. However, using live/dead stain alone for the determination of the appropriate chlorine dose against bulking may lead to an underestimation of the damage caused by chlorine to the useful microorganisms in the flocs. Indeed, using the live/dead stain, it was not easy to distinguish dead cells caused by chlorination from those originally present in the flocs The NA test was the most sensitive in detecting damage caused by chlorine to the floc-forming microorganisms. Therefore, for a safer determination of the chlorine dose effective against bulking and protective of the microbial activity of the sludge, the results of this study suggest coupling of the live/dead stain with the NA test and/or the OUR test.     

Characterisation of organic matter from anaerobic digestion of organic waste by aerobic microbial activity

The aim of this study was to examine whether the characterisation of organic matter on the basis of an oxygen uptake rate (OUR) could be applied to organic waste from an anaerobic waste treatment process. Three anaerobic digestion experiments were carried out in a bioreactor. Volatile fatty acids (VFA) and dissolved organic carbon (DOC) were monitored. OUR-experiments were carried out with diluted samples from the process. The graphs of the OUR-experiments showed a clear lag-phase, which was due to the slow adaptation of aerobic microorganisms. Model simulations of the OUR versus time curve showed sufficient agreement, if based on one fraction of readily biodegradable and two fractions of less easily biodegradable organic matter. The shape of the simulated graphs was affected considerably by the value of the maintenance energy requirement rate qm and could be improved by reducing the standard value qm = 1 d(-1) to qm = 0.1 d(-1). Only little agreement was achieved when comparing the results of the OUR-experiments with the VFA- and DOC-concentrations. Experiments with additional trace elements and vitamins led to an increase in the OUR and proved that the oxygen consumption was not exclusively determined by the availability of organic matter.     

On-line gas analysis in animal cell cultivation: I. Control of dissolved oxygen and pH

To monitor gas reaction rates in animal cell culture at constant dissolved oxygen concentration (DO) and constant pH it was necessary to develop improved control methods. Decoupling of both controllrs was obtained by manipulation of molar fractions of oxygen and carbon dioxide in the gas phase. Two pairs of DO and pH controllers were designed and tested both in simulation and exprimental runs. The first controller pair was developed for headspace aeration only, whereas the second controller pair was designed for bubble aeration using a microsparger and flushing the headspace with helium. pH was controlled by a conventional discrete PID controller in its velocity form. For DO control two linear state space feedback controllers with parameter adaptation were established. In these controllers the oxygen uptake rate (OUR) was considered as a disturbance and was not included in the mathematical model. The feedback gain adaptation was based on the difference between the actual molar fraction of oxygen at time step n and the initial molar fraction. This difference is related to OUR and was used to increase or decrease the state feedback controller gain (k and k(1), respectively) in a slow manner. With these controllers it was possible to get an excellent online estimate of OUR. In the case of bubble aeration a simple gas phase mass balance was sufficient, whereas during the headspace aeration a liquid phase balance was required. It has been shown that determination of OUR using gas balance requires a significantly better controller performance compared to just keeping DO and pH within reasonable limits. (c) 1995 John Wiley & Sons, Inc.     

How mutualisms arise in phytoplankton communities: building eco‐evolutionary principles for aquatic microbes

Extensive sampling and metagenomics analyses of plankton communities across all aquatic environments are beginning to provide insights into the ecology of microbial communities. In particular, the importance of metabolic exchanges that provide a foundation for ecological interactions between microorganisms has emerged as a key factor in forging such communities. Here we show how both studies of environmental samples and physiological experimentation in the laboratory with defined microbial co‐cultures are being used to decipher the metabolic and molecular underpinnings of such exchanges. In addition, we explain how metabolic modelling may be used to conduct investigations in reverse, deducing novel molecular exchanges from analysis of large‐scale data sets, which can identify persistently co‐occurring species. Finally, we consider how knowledge of microbial community ecology can be built into evolutionary theories tailored to these species’ unique lifestyles. We propose a novel model for the evolution of metabolic auxotrophy in microorganisms that arises as a result of symbiosis, termed the Foraging‐to‐Farming hypothesis. The model has testable predictions, fits several known examples of mutualism in the aquatic world, and sheds light on how interactions, which cement dependencies within communities of microorganisms, might be initiated.     

单细胞稳定同位素标记技术在固氮微生物中的应用研究

生物固氮是指固氮微生物将大气中氮气还原为生物可利用氨的过程,是环境中新氮的主要来源,调控初级生产力并影响氮储库的收支平衡。由于环境中大部分固氮微生物不可纯培养,不依赖培养且具有高空间分辨率水平的单细胞技术,成为研究固氮微生物的有力手段。~(15)N_2稳定同位素标记技术,以微生物对~(15)N的同化量或速率为依据,是表征微生物固氮活性的最直接手段。本文对~(15)N_2稳定同位素标记结合两种单细胞技术,即纳米二次离子质谱(Nano SIMS)和单细胞拉曼光谱,用于固氮微生物研究的最新进展进行了综述,内容包括揭示环境中高活性固氮微生物、空间分布、与其他生物的共生关系、细胞生理状态等,并进一步对近期发展的基于单细胞拉曼光谱的固氮微生物研究进行了展望。     

水圈微生物：推动地球重要元素循环的隐形巨人

正生活在水圈环境中的微生物数量巨大、遗传与代谢方式极为多样,它们驱动着地球上重要元素的循环。水圈微生物研究已经成为生命科学与地球科学的研究热点。国家自然科学基金委员会于2017年启动了"水圈微生物驱动地球元素循环的机制"重大研究计划(简称"水圈微生物"计划)。"水圈微生物"计划拟选择典型水圈生境,通过多学科交叉研究,借助新技术、新方法,揭示水圈微生物在物种、群落和生态水平驱动碳氮硫循环的机制及其环境响应,     

Limitation of oxygenic photosynthesis and oxygen consumption by phosphate and organic nitrogen in a hypersaline microbial mat: a microsensor study

Microbial mats are characterized by high primary production but low growth rates, pointing to a limitation of growth by the lack of nutrients or substrates. We identified compounds that instantaneously stimulated photosynthesis rates and oxygen consumption rates in a hypersaline microbial mat by following the short-term response (c. 6 h) of these processes to addition of nutrients, organic and inorganic carbon compounds, using microsensors. Net photosynthesis rates were not stimulated by compound additions. However, both gross photosynthesis and oxygen consumption were substantially stimulated (by a minimum of 25%) by alanine (1 mM) and glutamate (3.5 mM) as well as by phosphate (0.1 mM). A low concentration of ammonium (0.1 mM) did not affect photosynthesis and oxygen consumption, whereas a higher concentration (3.5 mM) decreased both process rates. High concentrations of glycolate (5 mM) and phosphate (1 mM) inhibited gross photosynthesis but not oxygen consumption, leading to a decrease of net photosynthesis. Photosynthesis was not stimulated by addition of inorganic carbon, nor was oxygen consumption stimulated by organic compounds like glycolate (5 mM) or glucose (5 mM), indicating that carbon was efficiently cycled within the mat. Photosynthesis and oxygen consumption were apparently tightly coupled, because stimulations always affected both processes to the same extent, which resulted in unchanged net photosynthesis rates. These findings illustrate that microsensor techniques, due to their ability to quantify all three processes, can clarify community responses to nutrient enrichment studies much better than techniques that solely monitor net fluxes.     

杂交瘤细胞培养中摄氧速率(OUR)的在线检测及其与细胞生长及代谢的关系

在批式及灌流培养条件下研究了杂交瘤细胞在无血清培养基中的生长、代谢情况与氧消耗的关系。应用动力学方法在线进行OUR的检测,同时离线取样检测其他参数。结果发现OUR与谷氨酰胺的消耗、抗体的生成及活细胞密度间有明显的相关关系,进一步的分析还发现在对数生长期,OUR与活细胞密度间具有良好的线性关系,qOUR(0.103±0.028)×10^-12mol/cell/h,可以通过它来进行细胞密度的在线检测。并通过以ΔOUR=0时刻作为灌流调整点进行连续灌流培养的初步实验验证了OUR作为培养过程反馈控制参数的可能性。     

Biodegradation of tannery wastewater using sequencing batch reactor--respirometric assessment

This investigation proved that respirometry combined with sequencing batch reactor (SBR) could be an effective way for the removal of COD in tannery wastewater. Measurement of oxygen uptake rates (OUR) and corresponding COD uptake rates showed that a 12-h operating cycle was optimum for tannery wastewater. The removal of COD by degradation was stoichiometric with oxygen usage. A plot of OUR values provided a good indication of the biological activity in the reactor. A high OUR value corresponded to the feed period; at the end of the cycle, when the substrate was depleted, the OUR value was low. At a 12-h SBR cycle with a loading rate of 1.9-2.1 kgm(-3) d(-1), removal of 80-82% COD, 78-80% TKN and 83-99% NH(3)-N were achieved. These removal efficiencies were much higher than the conventional aerobic systems. A simple method of COD fractionation was performed from the OUR and COD uptake rate data of the SBR cycle. About 66-70% of the influent COD was found to be readily biodegradable, 10-14% was slowly degradable and 17-21% was non-biodegradable. The oxygen mass transfer coefficient, K(L)a (19 +/- 1.7 h(-1)) was derived from respirometry. It was observed that with the exception of high organic load at the initial feed the oxygen transfer capacity was in excess of the OUR, and aerobic condition was generally maintained. Simultaneous nitrification-denitrification was observed in the SBR during the feed period as proved by mass balance.     

Microbial seed banks: the ecological and evolutionary implications of dormancy

Dormancy is a bet-hedging strategy used by a wide range of taxa, including microorganisms. It refers to an organism's ability to enter a reversible state of low metabolic activity when faced with unfavourable environmental conditions. Dormant microorganisms generate a seed bank, which comprises individuals that are capable of being resuscitated following environmental change. In this Review, we highlight mechanisms that have evolved in microorganisms to allow them to successfully enter and exit a dormant state, and discuss the implications of microbial seed banks for evolutionary dynamics, population persistence, maintenance of biodiversity, and the stability of ecosystem processes.