Microbial decolouration of azo dyes: A review

The textile industry is a substantial consumer of water and produces enormous volumes of contaminated water; the most important contaminants are azo dyes. Microbial processes for the treatment of textile wastewater have the advantage of being cost-effective and environmentally friendly and producing less sludge. The most promising microorganisms for wastewater treatment are those isolated from sites contaminated with dyes or from the sludge of treatment plants because they have adapted to survive in adverse conditions. The mechanism of microbial decolouration occurs from adsorption, enzymatic degradation or a combination of both. Both reductases and oxidases are involved in the microbial degradation process. The goal of microbial treatment is to decolourise and detoxify the dye-contaminated effluents. In this review, we summarise the methodologies used to evaluate the toxicity of azo dyes and their degradation products. Recent studies on the decolouration or degradation of azo dyes using algae, yeast, filamentous fungi and bacteria, genetically modified microorganisms, microbial consortia and microbiological systems combined with Advanced Oxidation Processes (AOPs) and Microbial Fuel Cells (MFCs) are discussed in this review.     

Microbiological contamination in genetically modified animals and proposals for a microbiological test standard for national universities in Japan.

The Biosafety Committee of the Japanese Association of Laboratory Animal Facilities of National Universities (JALAN) investigated recent episodes of microbiological contamination in genetically modified mice (GMM), and the countermeasures taken when the contaminated GMM were introduced into animal facilities, by questionnaires addressed to 53 animal facilities belonging to JALAN and serological tests. Although almost all of the contaminated GMM were accepted with conditions such as rederivation after or before reception and housing in designated rooms, contamination with a spectrum of microorganisms was demonstrated in GMM transferred domestically and from abroad. In serological tests, Mycoplasma pulmonis, mouse parvovirus, and mouse encephalomylitis virus were detected in GMM transferred from domestic facilities and from abroad. The present results of the questionnaires and serological tests suggest that GMM are highly and widely contaminated with microorganisms compared with mice from commercial breeders. Thus, we propose a microbiological requirement, including microbiological status--excellent, common, and minimum--as a guide for the transfer and procurement of mice and rats in Japan.     

Comparative genomics of "Dehalococcoides ethenogenes" 195 and an enrichment culture containing unsequenced "Dehalococcoides" strains

Tetrachloroethene (PCE) and trichloroethene (TCE) are prevalent groundwater contaminants that can be completely reductively dehalogenated by some "Dehalococcoides" organisms. A Dehalococcoides-organism-containing microbial consortium (referred to as ANAS) with the ability to degrade TCE to ethene, an innocuous end product, was previously enriched from contaminated soil. A whole-genome photolithographic microarray was developed based on the genome of "Dehalococcoides ethenogenes" 195. This microarray contains probes designed to hybridize to >99% of the predicted protein-coding sequences in the strain 195 genome. DNA from ANAS was hybridized to the microarray to characterize the genomic content of the ANAS enrichment. The microarray results revealed that the genes associated with central metabolism, including an apparently incomplete carbon fixation pathway, cobalamin-salvaging system, nitrogen fixation pathway, and five hydrogenase complexes, are present in both strain 195 and ANAS. Although the gene encoding the TCE reductase, tceA, was detected, 13 of the 19 reductive dehalogenase genes present in strain 195 were not detected in ANAS. Additionally, 88% of the genes in predicted integrated genetic elements in strain 195 were not detected in ANAS, consistent with these elements being genetically mobile. Sections of the tryptophan operon and an operon encoding an ABC transporter in strain 195 were also not detected in ANAS. These insights into the diversity of Dehalococcoides genomes will improve our understanding of the physiology and evolution of these bacteria, which is essential in developing effective strategies for the bioremediation of PCE and TCE in the environment.     

Database on natural and transgenic luminous microorganisms: "BiolumBase"

The database "BiolumBase" is designed for the selection and systematization of available world information on microorganisms containing bioluminescent systems; it includes two sections: "natural" and "transgenic" luminous microorganisms. At present, logic schemes of divisions, classification of the objects, presentation of characteristics, and the inputs of relative information, as well as the necessary program modules including links to the database, are developed. The database is constructed on the basis of published data and our own experimental results; the subsequent linkage of the database to the Internet is envisaged. Users will be able to obtain not only the catalogues of strains but also information concerning the properties and functions of the known species of luminous bacteria, the structure, regulatory mechanisms, and application of bioluminescent systems and genetically engineered constructions with lux genes, as well as to find references and to search strains by using any set of attributes. The database will provide information that is of interest for the development of microbial ecology and biotechnology, in particular, for the prediction of biological hazard from the application of transgenic strains.     

Influence of introducing the genetically modified strain <Emphasis Type="Italic">Sinorhizobium meliloti</Emphasis> ACH-5 on the structure of the soil microbial community

To study the influence of genetically modified microorganisms (GMM) on the number and structure of the soil microbial community, we introduced the genetically modified strain of Sinorhizobium meliloti into soil under controlled laboratory conditions. The analysis of the dynamics of soil microorganisms of all the main groups (archaea, bacteria, fungi) using the PCR with real-time detection and the analysis of the species structure of all the indicated components using T-RFLP were carried out for a month. The results of the quantitative PCR demonstrated that none of the components of the soil microbial community was appreciably influenced by the GMM introduced. The number of GMM decreased over a month more than 300-fold. The analysis of the dynamics of the eubacterial, archaeal, and fungal communities using T-RFLP did not detect fundamental changes in their structure.     

High genetic variation among closely related red oak (<Emphasis Type="Italic">Quercus rubra</Emphasis>) populations in an ecosystem under metal stress: analysis of gene regulation

Metal toxicity is a major abiotic stressor of plants. It has been established that changes in genetic variation occur very rapidly in plants in response to environmental stressors such as increased levels of metals. Quercus rubra (red oak) is a pioneer species in mining regions contaminated with metals in Northern Ontario (Canada). The objectives of the study were to (1) determine the level of genetic variation in Q. rubra populations from mining damaged ecosystems using RAPD marker system and (2) assess the level of gene expression of candidate genes for nickel resistance. Total gene diversity (H_T) and the mean gene diversity among populations (H_S) were 0.22 and 0.19, respectively. The percent of polymorphic loci within populations was high ranging from 61 % (Capreol) to 72 % (Daisy Lake) despite a high level of gene flow (2.4). The population differentiation (G_ST) value was low (0.17). No significant difference was found among the contaminated and reference sites for all the genetic parameters estimated. Hence, all the Q. rubra populations from the metal-contaminated and damaged ecosystems are genetically sustainable. Moreover, this study reveals that all populations were genetically closely related with genetic distance values varying from 0.17 to 0.35. A zinc finger protein of Arabidopsis thaliana (ZAT11) gene involved in nickel resistance was differentially expressed in samples analyzed. There was a 120 times higher of ZAT11 expression in samples from metal contaminated areas of Wahnapitae Dam compared to other metal contaminated and uncontaminated sites, but no association between soil metal levels and expression of ZAT11 was established.     

Biodegradation of methyl <Emphasis Type="Italic">tert</Emphasis>-butyl ether using bacterial strains

Prospective methyl tert-butyl ether (MTBE) degrading bacterial strains and/or consortia were identified. The potential for aerobic degradation of MTBE was examined using bacterial isolates from contaminated soils and groundwater. Using the 16S rDNA protocol, two isolates capable of degrading MTBE (Rhodococcus pyridinivorans 4A and Achromobacter xylosoxidans 6A) were identified. The most efficient consortium of microorganisms was acquired from contaminated groundwater. The growth of both strains and the consortium on MTBE was supported by various organic substrates, and monitored using Bioscreen®. The biochemical oxygen demand of the cultures was measured using OxiTop®, and their MTBE concentrations were estimated by gas chromatography. After 3 weeks of aerobic cultivation using n-alkanes as cosubstrate, the concentration of MTBE in R. pyridinivorans 4A was reduced to 62.4 % of its initial amount (50 ppm).     

Synergistic plant-microbes interactions in the rhizosphere: a potential headway for the remediation of hydrocarbon polluted soils

Soil pollution is an unavoidable evil; many crude-oil exploring communities have been identified to be the most ecologically impacted regions around the world due to hydrocarbon pollution and their concurrent health risks. Several clean-up technologies have been reported on the removal of hydrocarbons in polluted soils but most of them are either very expensive, require the integration of advanced mechanization and/or cannot be implemented in small scale. However, “Bioremediation” has been reported as an efficient, cost-effective and environment-friendly technology for clean-up of hydrocarbon”s contaminated soils. Here, we suggest the implementation of synergistic mechanism of bioremediation such as the use of rhizosphere mechanism which involves the actions of plant and microorganisms, which involves the exploitation of plant and microorganisms for effective and speedy remediation of hydrocarbon”s contaminated soils. In this mechanism, plant”s action is synergized with the soil microorganisms through the root rhizosphere to promote soil remediation. The microorganisms benefit from the root metabolites (exudates) and the plant in turn benefits from the microbial recycling/solubilizing of mineral nutrients. Harnessing the abilities of plants and microorganisms is a potential headway for cost-effective clean-up of hydrocarbon”s polluted sites; such technology could be very important in countries with great oil producing activities/records over many years but still developing.     

Biosynthesis of <Emphasis Type="SmallCaps">d</Emphasis>-lactic acid from lignocellulosic biomass

d-lactic acid is a versatile and important industrial chemical that can be applied in the synthesis of thermal-resistant poly-lactic acid. Biosynthesis of d-lactic acid can be achieved by a variety of microorganisms, including lactic acid bacteria, yeast, and fungi; however, the final product yield, optical purity, and the utilization of both glucose and xylose are restricted. Consequently, engineered microbial systems are essential to attain high titer, productivity, and complete utilization of sugars. Herein, we critically evaluate the promising wild-type microorganisms, as well as genetically modified microorganisms to produce enantiomerically pure d-lactic acid, particularly from renewable lignocellulosic biomass. In addition, innovative bioreactor operation, metabolic flux analysis, and recent genetic engineering methods for targeted microbial d-lactic acid synthesis will be discussed.     

PCE dechlorination by non-<Emphasis Type="Italic">Dehalococcoides</Emphasis> in a microbial electrochemical system

The bioremediation of tetrachloroethene (perchloroethene; PCE) contaminated sites generally requires a supply of some fermentable organic substrates as an electron donor. On the other hand, organic substrates can induce the massive growth of microorganisms around the injection wells, which can foul the contaminated subsurface environment. In this study, PCE dechlorination to ethene was performed in a microbial electrochemical system (MES) using the electrode (a cathode polarized at ?500 mV vs. standard hydrogen electrode) as the electron donor. Denaturing gel gradient electrophoresis and pyrosequencing revealed a variety of non-Dehalococcoides bacteria dominant in MES, such as Acinetobacter sp. (25.7 % for AS1 in suspension of M3), Rhodopseudomonas sp. (10.5 % for AE1 and 10.1 % for AE2 in anodic biofilm of M3), Pseudomonas aeruginosa (22.4 % for BS1 in suspension of M4), and Enterobacter sp. (21.7 % for BE1 in anodic biofilm of M4) which are capable of electron transfer, hydrogen production and dechlorination. The Dehalococcoides group, however, was not detected in this system. Therefore, these results suggest that a range of bacterial species outside the Dehalococcoides can play an important role in the microbial electrochemical dechlorination process, which may lead to innovative bioremediation technology.     

Methods for detecting recombinant DNA in the environment

The successful introduction of genetically modified and genetically engineered microorganisms into the environment requires a quantitative evaluation of the survival and dispersion of the microorganisms and specific gene(s) in the environment. The objective of this article is to examine the applicability, suitability, and significance of existing and new methods for detecting and monitoring the recombinant genes or organisms introduced into the environment. Conventional microbiological method(s) involving the selective and differential growth of microorganism(s) adn other quantitative approaches such as the most-probable-number (MPN) method and direct microscopic observation (e.g., acridine orange direct count analysis) have drawbacks and are not specific or universally applicable. Direct enumeration by immunofluorescence by the use of fluorescent dye seems more sensitive although still not perfect. However, the molecular methodologies such as the use of gene probes, plasmid epidemiology, antibiotic resistant marker strains, and protein electrophoresis and bacteriophage sensitivity are receiving more attention. As yet, the technology of DNA:DNA hybridization appears to be very useful, sensitive, and accurate for detecting and monitoring the microorganisms in the environment, although improvements are required. New approaches can be developed which may include biochemical signature compounds as well as gene cassettes to be used in a complementary fashion with conventional and molecular techniques for quantifying specific genotypes and genes in the environment.     

A critical review of the application of white rot fungus to environmental pollution control

Research on white rot fungi for environmental biotechnology has been conducted for more than 20 years. In this article, we have reviewed processes for cell growth and enzyme production including the factors influencing enzyme productivity and the methods for enhancement of enzyme production. Significant progress has been achieved in molecular biology related to white rot fungi, especially related to the extraction of genetic material (RNA and DNA), gene cloning and the construction of genetically engineered microorganisms. The development of biotechnologies using white rot fungi for environmental pollution control has been implemented to treat various refractory wastes and to bioremediate contaminated soils. The current status and future research needs for fundamentals and application are addressed in this review.     

Utility of routine evaluation of sterility of cellular therapy products with or without extensive manipulation: Best practices and clinical significance

Background

We analyzed the results of routine sterility testing performed in our center over the last 10 years, in the context both hematopoietic stem cell transplantation (HSCT) and Advanced Therapeutic Medicinal Products (ATMPs).

生物组学在污染环境微生物修复研究中的应用

随着分子生物学、生物信息学和各种理化检测技术的发展,特别是人类基因组计划成功实施以来,基因组学研究取得了重大突破与进展。而包括转录组学、蛋白组学和代谢组学在内的后基因组学也相继出现,并被广泛应用在环境微生物学的各个研究领域。本文主要概述了当前基因组学、转录组学、蛋白质组学和代谢组学在污染环境生物修复研究中的最新研究进展,分析比较了各组学的优势与不足,同时结合本课题组的主要研究方向探讨了各生物组学在赤潮生消过程和有机污染物降解机理等研究中的应用。     

Emulsification of hydrocarbons by subsurface bacteria

Summary Biosurfactants have potential for use in enhancement of in situ biorestoration by increasing the bioavailability of contaminants. Microorganisms isolated from biostimulated, contaminated and uncontaminated zones at the site of an aviation fuel spill and hydrocarbon-degrading microorganisms isolated from sites contaminated with unleaded gasoline were examined for their abilities to emulsify petroleum hydrocarbons. Emulsifying ability was quantified by a method involving agitation and visual inspection. Biostimulated-zone microbes and hydrocarbon-degrading microorganisms were the best emulsifiers as compared to contaminated and uncontaminated zone microbes. Biostimulation (nutrient and oxygen addition) may have been the dominant factor which selected for and encouraged growth of emulsifiers; exposure to hydrocarbon was also important. Biostimulated microorganisms were better emulsifiers of aviation fuel (the contaminant hydrocarbon) than of heavier hydrocarbon to which they were not previously exposed. By measuring surface tension changes of culture broths, 11 out of 41 emulsifiers tested were identified as possible biosurfactant producers and two isolates produced large surface tension reductions indicating the high probability of biosurfactant production.     

Genetically Distinct Populations of the Dinoflagellate <Emphasis Type="Italic">Peridinium limbatum</Emphasis> in Neighboring Northern Wisconsin Lakes

The extent to which free-living microorganisms exist in geographically isolated, genetically distinct populations is a subject of continuing debate. Some authorities contend that many microorganisms have cosmopolitan distributions, while others provide evidence that more limited geographical distribution of genetically distinct populations can occur. We report the occurrence of two morphologically similar, but genetically distinct, populations of the microbial eukaryote Peridinium limbatum (Stokes) Lemmermann from neighboring Northern Wisconsin freshwater bodies. Five strains of P. limbatum were cultured by single-cell isolation from both Crystal Lake and Crystal Bog (Oneida Co., WI). Genetic variation between the two populations encompassed 8.9% (mean of 35.4 of 397 nucleotides) of the nuclear ribosomal DNA internal transcribed spacer (ITS1 and ITS2) region. In contrast, 0.5% (mean of 2.25 of 397 nucleotides) variation was observed within the Crystal Lake population and 0.3% (mean of 1.21 of 397 nucleotides), within the Crystal Bog population. This difference between the two populations was highly statistically significant (p-value << 0.001). The extent of genetic variation between the two P. limbatum populations was greater than that reported in the literature for some morphologically distinguishable microalgal species, suggesting the occurrence of cryptic sister species. On the other hand, hybrid sequences obtained from one of the Crystal Lake strains suggest that the two populations may still be members of a single sexually compatible biological species. Our data suggest that the two neighboring P. limbatum populations may be diverging genetically under conditions of limited gene flow, suggesting a mechanism for the origin of geographically isolated, genetically distinct populations of microbial eukaryotes.     

微生物分子生态学技术及其在环境污染研究中的应用

较为系统地概述了核酸探针检测技术、利用引物的PCR技术、DNA序列分析技术和电泳分离及显示技术在国内外的研究进展,并探讨了这些技术在环境污染研究中的应用及其方向。结果表明,这些被认为是重要的微生物分子生态学技术,在探索微生物与污染环境之间的相互关系中发挥了重要作用。促进了污染环境的微生物遗传适应进化机制的研究,污染物的微生物降解有关基因的定位及微生物工程菌的构建等方面的工作,从而推进了污染环境微生物修复的分子生态学的发展。     

<Emphasis Type="Italic">In-silico</Emphasis> identification of phenotype-biased functional modules

Background

Phenotypes exhibited by microorganisms can be useful for several purposes, e.g., ethanol as an alternate fuel. Sometimes, the target phenotype maybe required in combination with other phenotypes, in order to be useful, for e.g., an industrial process may require that the organism survive in an anaerobic, alcohol rich environment and be able to feed on both hexose and pentose sugars to produce ethanol. This combination of traits may not be available in any existing organism or if they do exist, the mechanisms involved in the phenotype-expression may not be efficient enough to be useful. Thus, it may be required to genetically modify microorganisms. However, before any genetic modification can take place, it is important to identify the underlying cellular subsystems responsible for the expression of the target phenotype.

Results

In this paper, we develop a method to identify statistically significant and phenotypically-biased functional modules. The method can compare the organismal network information from hundreds of phenotype expressing and phenotype non-expressing organisms to identify cellular subsystems that are more prone to occur in phenotype-expressing organisms than in phenotype non-expressing organisms. We have provided literature evidence that the phenotype-biased modules identified for phenotypes such as hydrogen production (dark and light fermentation), respiration, gram-positive, gram-negative and motility, are indeed phenotype-related.

Conclusion

Thus we have proposed a methodology to identify phenotype-biased cellular subsystems. We have shown the effectiveness of our methodology by applying it to several target phenotypes. The code and all supplemental files can be downloaded from (http://freescience.org/cs/phenotype-biased-biclusters/).