SEARCH · PubMed Health

Results for “Microbial community structure”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 397 records · Page 22Linked to original sources

PubMed2001

A strategy for optimizing quality and quantity of DNA extracted from soil.

The efficiency of a bead beating method was studied in detail with regard to a variety of factors including beating time and speed, volume and temperature of the buffer, as well as amount and type of beads employed. The results presented here reveal that all of these parameters have a significant effect on yield and quality of DNA extracted from soils. Precise adjustment of extraction conditions allows for significantly higher yields of high quality DNA from soils than previously reported. We further evaluated the effect of the extraction conditions on the apparent soil microbial community structures, as observed by polymerase chain reaction (PCR) and RFLP. Differences in the fingerprints of DNA extracted under different conditions suggest that results could be biased when using gentle extraction procedures. Based on multiple subsequent extractions using very harsh extraction conditions, we propose a protocol for the quantification of the total DNA content in soils. Extractions from six soils of different texture and chemical characteristics with selected bead beating protocols revealed that the quality (fragment size and purity) of the extracted DNA was generally very good, but also depended on the soil characteristics. While a single, general protocol for optimal DNA recovery from all soils cannot be given, this study provides detailed guidelines on how to optimize the general method to obtain optimal DNA from individual soils.

Buffers↗

PubMed2000

Anaerobic decomposition of halogenated aromatic compounds.

Halogenated compounds constitute one of the largest groups of environmental pollutants, partly as a result of their widespread use as biocides, solvents and other industrial chemicals. A critical step in degradation of organohalides is the cleavage of the carbon?halogen bond. Reductive dehalogenation is generally the initial step in metabolism under methanogenic conditions, which requires a source of reducing equivalents, with the halogenated compound serving as an electron acceptor. Dehalogenation is greatly influenced by alternate electron acceptors; e.g. sulfate frequently inhibits reductive dehalogenation. On the other hand, a number of halogenated aromatic compounds can be degraded under different electron-accepting conditions and their complete oxidation to CO(2) can be coupled to processes such as denitrification, iron(III)-reduction, sulfate reduction and methanogenesis. Reductive dehalogenation was the initial step in degradation not only under methanogenic, but also under sulfate- and iron(III)-reducing conditions. Dehalogenation rates were in general slower under sulfidogenic and iron-reducing conditions, suggesting that dehalogenation was affected by the electron acceptor. The capacity for dehalogenation appears to be widely distributed in anoxic environments; however, the different substrate specificities and activities observed for the halogenated aromatic compounds suggest that distinct dehalogenating microbial populations are enriched under the different reducing conditions. Characterization of the microbial community structure using a combination of biomolecular techniques, such as cellular fatty acid profiling, and 16 S rRNA fingerprinting/sequence analysis, was used to discern the distinct populations enriched with each substrate and under each electron-accepting condition. These combined techniques will aid in identifying the organisms responsible for dehalogenation and degradation of halogenated aromatic compounds.

Journal Article↗

PubMed2000

Molecular evidence for genus level diversity of bacteria capable of catalyzing anaerobic ammonium oxidation.

Recently, a bacterium capable to oxidize ammonium anaerobically at a high rate was identified as novel member of the Planctomycetales (Strous, M., Fuersi, J. A., Kramer, E. H. M., Logemann, S., Muyzer, G., van de Pas-Schoonen, K. T., Webb, R. I., Kufnen, J. G., and Jetten, M. S. M.: Nature 400, 446-449, 1999). Here we investigated the microbial community structure of a trickling filter biofilm with a high anaerobic ammonium oxidation activity. Fluorescence in situ hybridization (FISH) with a set of nine probes designed for specific identification of the recently described anaerobic ammonium oxidizer demonstrated that only one probe hybridized to bacteria within the biofilm. For phylogenetic characterization of putative biofilm anaerobic ammonium oxidizers a full-cycle 16S rDNA approach was performed by using a Planctomycetales-specific forward primer for PCR amplification. Of the twenty-five 16S rDNA fragments (1364 bp in length) amplified from the biofilm, nine were affiliated to the Planctomycetales. Comparative analysis showed that these sequences were more than 98.9% similar to each other but only distantly related to the previously recognized anaerobic ammonium oxidizer (below 91% similarity) and all other organisms represented in public 16S rRNA databases (similarities of below 79%). The retrieved sequences and the previously recognized anaerobic ammonium oxidizer represent two well-separated groups of a deep-branching lineage within the Planctomycetales. Quantitative FISH analysis with a newly designed specific probe showed that the novel bacterium, provisionally classified as "Candidatus Kuenenia stuttgartiensis" constituted the dominant fraction of the biofilm bacteria. In situ probing revealed that ammonia-oxidizing bacteria of the beta-subclass of Proteobacteria were also present, albeit in significant smaller amounts, within the anoxic biofilm. Comparative sequence analysis of a stretch of the gene encoding ammonia-monooxygenase (amoA) demonstrated the occurrence of the DNA of at least three different populations of beta-subclass ammonia oxidizers within the biofilm.

Anaerobiosis↗

PubMed2000

Engineering bacteria for bioremediation.

The treatment of environmental pollution by microorganisms is a promising technology. Various genetic approaches have been developed and used to optimize the enzymes, metabolic pathways and organisms relevant for biodegradation. New information on the metabolic routes and bottlenecks of degradation is still accumulating, enlarging the available toolbox. With molecular methods allowing the characterization of microbial community structure and activities, the performance of microorganisms under in situ conditions and in concert with the indigenous microflora will become predictable.

Bacteria↗

PubMed2001

Effect of repeated addition of nitrite to semi-continuous activated sludge reactors.

To a semi-continuous lab-scale activated sludge system (SCAS), nitrite was dosed discontinuously, i.e. together with the feed. The nitrite was added at a concentration (50 mg N/l) which had been found earlier not to cause acute inhibition of the general running of the reactors. Repeated nitrite addition, however, exerted an inhibitory effect which was reflected in decreased removal efficiencies and was also expressed in decreases in total respiratory and nitrifying activity of 20% and 40%, respectively. The inhibition by nitrite was reversible as removal efficiencies improved after ceasing nitrite addition. The two consecutive test runs revealed that addition of nitrite gave rise to high amounts of dispersed cells and free-swimming protozoa. Moreover, DGGE patterns confirmed a shift in microbial community structure upon application of nitrite.

Bioreactors↗

PubMed2002

A review and update of the microbiology of enhanced biological phosphorus removal in wastewater treatment plants.

Enhanced biological phosphorus removal (EBPR) from wastewater can be more-or-less practically achieved but the microbiological and biochemical components are not completely understood. EBPR involves cycling microbial biomass and influent wastewater through anaerobic and aerobic zones to achieve a selection of microorganisms with high capacity to accumulate polyphosphate intracellularly in the aerobic period. Biochemical or metabolic modelling of the process has been used to explain the types of carbon and phosphorus transformations in sludge biomass. There are essentially two broad-groupings of microorganisms involved in EBPR. They are polyphosphate accumulating organisms (PAOs) and their supposed carbon-competitors called glycogen accumulating organisms (GAOs). The morphological appearance of microorganisms in EBPR sludges has attracted attention. For example, GAOs as tetrad-arranged cocci and clusters of coccobacillus-shaped PAOs have been much commented upon and the use of simple cellular staining methods has contributed to EBPR knowledge. Acinetobacter and other bacteria were regularly isolated in pure culture from EBPR sludges and were initially thought to be PAOs. However, when contemporary molecular microbial ecology methods in concert with detailed process performance data and simple intracellular polymer staining methods were used, a betaproteobacteria called 'Candidatus Accumulibacter phosphatis' was confirmed as a PAO and organisms from a novel gammaproteobacteria lineage were GAOs. To preclude making the mistakes of previous researchers, it is recommended that the sludge 'biography' be well understood--i.e. details of phenotype (process performance and biochemistry) and microbial community structure should be linked.

Betaproteobacteria↗

PubMed2007

Microbial landscapes: new paths to biofilm research.

It is the best of times for biofilm research. Systems biology approaches are providing new insights into the genetic regulation of microbial functions, and sophisticated modelling techniques are enabling the prediction of microbial community structures. Yet it is also clear that there is a need for ecological theory to contribute to our understanding of biofilms. Here, we suggest a concept for biofilm research that is spatially explicit and solidly rooted in ecological theory, which might serve as a universal approach to the study of the numerous facets of biofilms.

Bacterial Physiological Phenomena↗

PubMed2003

Microbial characterization of the Mars Odyssey spacecraft and its encapsulation facility.

Microbial characterization of the Mars Odyssey spacecraft and the Kennedy Space Center Spacecraft Assembly and Encapsulation Facility II (SAEF-II) was carried out by both culture-based and molecular methods. The most dominant cultivable microbes were species of Bacillus, with comamonads, microbacteria and actinomycetales also represented. Several spore-forming isolates were resistant to gamma-radiation, UV, H2O2 and desiccation, and one Acinetobacter radioresistens isolate and several Aureobasidium, isolated directly from the spacecraft, survived various conditions. Sequences arising in clone libraries were fairly consistent between the spacecraft and facility; predominant genera included Variovorax, Ralstonia and Aquaspirillum. This study improves our understanding of the microbial community structure, diversity and survival capabilities of microbes in an encapsulation facility and physically associated with colocated spacecraft.

Actinomycetales↗

PubMed2025

Diversity of leaf- and root microbiomes among genotypes and market classes of desert-grown lettuce (Lactuca sativa L.).

Plant microbiomes are increasingly acknowledged both as extensions of plant characteristics and as biological factors that influence plant traits important for nutrition and resilience. In the context of global change, manipulation of microbiomes has the potential to complement genetic approaches to enhance crop health and productivity under rising heat and drought stress. Understanding the factors that influence microbial communities and their variation across plant genotypes is essential for developing such capabilities. We employed metabarcoding via the Illumina sequencing platform to investigate microbial communities that occur within healthy leaves and roots of 12 lettuce genotypes (Lactuca sativa L.) grown in a desert agriculture environment. We detected diverse foliar- and root-endophytic fungi and bacteria in field-grown lettuce at the Maricopa Agricultural Center (Arizona, USA). The composition of microbial community structure varied with foliar chemistry and root traits. Notably, levels of zinc and other beneficial nutrients in the leaves were strongly linked with specific endophytes. These results document the lettuce microbiome in desert farming and provide insights into endophytes in lettuce leaves, which are noteworthy because they remain after washing and are regularly ingested.

Lactuca↗

PubMed2004

Molecular ecological analysis of the gastrointestinal microbiota: a review.

The gastrointestinal (GI) microbiota of mammals is characterized by its high population density, wide diversity and complexity of interactions. While all major groups of microbes are represented, bacteria predominate. Importantly, bacterial cells outnumber animal (host) cells by a factor of ten and have a profound influence on nutritional, physiological and immunological processes in the host animal. Our knowledge of the molecular and cellular bases of host-microbe interactions is limited, though critically needed to determine if and how the GI microbiota contributes to various enteric disorders in humans and animals. Traditionally, GI bacteria have been studied via cultivation-based techniques, which are labor intensive and require previous knowledge of individual nutritional and growth requirements. Recently, findings from culture-based methods have been supplemented with molecular ecology techniques that are based on the 16S rRNA gene. These techniques enable characterization and quantification of the microbiota, while also providing a classification scheme to predict phylogenetic relationships. The choice of a particular molecular-based approach depends on the questions being addressed. Clone libraries can be sequenced to identify the composition of the microbiota, often to the species level. Microbial community structure can be analyzed via fingerprinting techniques, while dot blot hybridization or fluorescent in situ hybridization can measure abundance of particular taxa. Emerging approaches, such as those based on functional genes and their expression and the combined use of stable isotopes and biomarkers, are being developed and optimized to study metabolic activities of groups or individual organisms in situ. Here, a critical summary is provided of current molecular ecological approaches for studying the GI microbiota.

Digestive System↗

PubMed2005

Microbial indices of soil fertility.

AIMS: To find the new microbial parameters explaining the soil fertility from the microbial community viewpoint. METHODS AND RESULTS: Fatty acid methyl ester (FAME) analysis and terminal-restriction fragment length polymorphism (T-RFLP) analysis were carried out using 16 differently treated plots from the same field that had been kept under different fertilizer management systems since 1984. It was found that organic fertilizer application had small impact, whereas chemical fertilizer application, especially ammonium-nitrogen fertilizer, had strong impact on microbial community structures. Principal component analysis was conducted based on soil chemical and physical parameters, crop yields, FAMEs and terminal-restriction fragments (T-RFs) to provide 10 FAMEs and 10 T-RFs showing strong relation with soil fertility. CONCLUSION: We defined these 10 FAMEs and 10 T-RFs as 'keystone' biological parameters explaining soil fertility in the soil. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first report on the FAMEs and T-RFs related to soil fertility. Both analyses are rapid, inexpensive and reproducible means. As field assessment needs precise and rapid analysis, FAME and T-RFLP analyses and these new parameters are very useful to analyse soil fertility at biological viewpoint.

Agriculture↗

PubMed2006

Acidophilic microbial communities associated with a natural, biodegraded hydrocarbon seepage.

Characterization of microbial communities present in a surface petroleum seep in which hydrocarbons have been biodegraded for thousands of years in order to improve the understanding on natural petroleum biodegradation. DNA was extracted from a natural, surface petroleum seep and subjected to culture independent analysis (rRNA gene-based denaturing gradient gel electrophoresis and phylogenetic analysis of clone libraries). Molecular analysis suggested dominance by acidophilic bacteria, especially Alphaproteobacteria (mainly bacteria related to Acidiphilium and Acidocella). Archaea were not detected, but fungi were present. pH of the samples was around 3.5. Acidophilic microbial communities are associated with an acidic petroleum seep. Microbial community structure analysis gives information on the environmental conditions under which petroleum biodegradation occurs. This knowledge could be applied to define conditions for specific cultivation or activity measurements. The activity of acidophilic micro-organisms deserves more attention with respect to their involvement in natural petroleum degradation. This knowledge will contribute to the design of oil bioremediation strategies for polluted acidic settings.

Acidiphilium↗

PubMed2004

Formation of granules and Methanosaeta fibres in an anaerobic migrating blanket reactor (AMBR).

It has generally been accepted that the formation of granules in anaerobic wastewater treatment systems requires a hydraulic upflow pattern. To evaluate this hypothesis, we operated an anaerobic migrating blanket reactor (AMBR) without a hydraulic upflow pattern, using a synthetic wastewater containing acetate, propionate, butyrate and sucrose. We provided conditions amenable to the formation of granules by operating the system with a moderate hydraulic selection pressure, which in this system was not the result of a hydraulic upflow pattern, but was provided by migration of biomass and intermittent mechanical mixing. Granules were first noticed after 2 months of operation, and it took another 2 months for a mature granular blanket to develop. Besides granules, approximately 1-cm-long Methanosaeta fibres developed and, after 6 months of operation, 30% of biomass consisted of these fibres. Quantitative membrane hybridization showed that almost all the total 16S rRNA extracted from fibres consisted of 16S rRNA from Methanosaeta concilii. This finding indicates that it was possible to develop pockets consisting almost entirely of an organism with a very limited substrate utilization spectrum (only acetate) in a system that was fed a synthetic wastewater containing acetate, propionate, butyrate and sucrose and that is known for its ability to develop biomass with a complex microbial community structure.

Anaerobiosis↗

PubMed2005

Loss of diversity of ammonia-oxidizing bacteria correlates with increasing salinity in an estuary system.

Ammonia-oxidizing bacteria (AOB) play an important role in nitrogen cycling in estuaries, but little is known about AOB diversity, distribution and activity in relation to the chemical and physical changes encountered in estuary systems. Although estuarine salinity gradients are well recognized to influence microbial community structure, few studies have examined the influence of varying salinity on the diversity and stability of AOB populations. To investigate these relationships, we collected sediment samples from low-, mid- and high-salinity sites in Plum Island Sound estuary, MA, during spring and late summer over 3 years. Ammonia-oxidizing bacteria distribution and diversity were assessed by terminal restriction fragment length polymorphism (TRFLP) analysis of the ammonia monooxygenase (amoA) gene, and fragments were identified by screening amoA clone libraries constructed from each site. Most striking was the stability and low diversity of the AOB community at the high-salinity site, showing little variability over 3 years. Ammonia-oxidizing bacteria at the high-salinity site were not closely related to any cultured AOB, but were most similar to Nitrosospira spp. Ammonia-oxidizing bacteria at the mid- and low-salinity sites were distributed among Nitrosospira-like sequences and sequences related to Nitrosomonas ureae/oligotropha and Nitrosomonas sp. Nm143. Our study suggests that salinity is a strong environmental control on AOB diversity and distribution in this estuary.

Ammonia↗

PubMed2006

Statistical methods for characterizing diversity of microbial communities by analysis of terminal restriction fragment length polymorphisms of 16S rRNA genes.

The analysis of terminal restriction fragment length polymorphisms (T-RFLP) of 16S rRNA genes has proven to be a facile means to compare microbial communities and presumptively identify abundant members. The method provides data that can be used to compare different communities based on similarity or distance measures. Once communities have been clustered into groups, clone libraries can be prepared from sample(s) that are representative of each group in order to determine the phylogeny of the numerically abundant populations in a community. In this paper methods are introduced for the statistical analysis of T-RFLP data that include objective methods for (i) determining a baseline so that 'true' peaks in electropherograms can be identified; (ii) a means to compare electropherograms and bin fragments of similar size; (iii) clustering algorithms that can be used to identify communities that are similar to one another; and (iv) a means to select samples that are representative of a cluster that can be used to construct 16S rRNA gene clone libraries. The methods for data analysis were tested using simulated data with assumptions and parameters that corresponded to actual data. The simulation results demonstrated the usefulness of these methods in their ability to recover the true microbial community structure generated under the assumptions made. Software for implementing these methods is available at http://www.ibest.uidaho.edu/tools/trflp_stats/index.php.

Bacteria↗

PubMed2006

Candida biofilms on implanted biomaterials: a clinically significant problem.

In recent years there has been an increasing appreciation that microbial biofilms are ubiquitous, which has resulted in a number of studies on infectious diseases from a biofilm perspective. Biofilms are defined as structured microbial communities that are attached to a surface and encased in a matrix of exopolymeric material. A wide range of biomaterials used in clinical practice have been shown to support colonization and biofilm formation by Candida spp., and the increase in Candida infections in the last decades has almost paralleled the increase and widespread use of a broad range of medical implant devices, mainly in populations with impaired host defenses. Formation of Candida biofilms has important clinical repercussions because of their increased resistance to antifungal therapy and the ability of cells within biofilms to withstand host immune defenses. Further recognition and understanding of the role of Candida biofilms in human infection should help in the clinical management of these recalcitrant infections.

Biofilms↗

PubMed2000

Life at the oxic-anoxic interface: microbial activities and adaptations.

Molecular oxygen is one of the most important reactants in biogeochemical cycles. Due to its low solubility in water, the consumption of oxygen leads to the development of oxic-anoxic interfaces, which separate aerobic from anaerobic processes in virtually all environments, ranging in scale from oceanic sediments to the fecal pellets of a small soil invertebrate. Three case studies were selected to illustrate the basic situation and the specific characteristics of oxic-anoxic interfaces: sediments, the rhizosphere of aquatic plants, and the intestinal tract of insects. Each system is governed by the same general principles, but striking differences arise from, e.g., the nature of the major microbial activities and the mechanisms controlling metabolite fluxes. Also scale and dimensional differences as well as the consequences of temporal fluctuations are of fundamental importance. Recent developments in microbial ecology, which often combine traditional and modern approaches, have significantly furthered our understanding of the specific microniches and the metabolic and behavioral adaptations of microorganisms to life at the oxic-anoxic interface. New concepts help to define the targets of future studies: the spatial organization of microbial populations, their microenvironments and in situ activities, and the functional interactions within structured microbial communities.

Adaptation, Physiological↗

PubMed2025

Expansion of a bacterial operon during cancer treatment ameliorates fluoropyrimidine toxicity.

Dose-limiting toxicities remain a major barrier to drug development and therapy, revealing the limited predictive power of human genetics. Here, we demonstrate the utility of a more comprehensive approach to studying drug toxicity through longitudinal profiling of the human gut microbiome during colorectal cancer (CRC) treatment (NCT04054908) coupled to cell culture and mouse experiments. Substantial shifts in gut microbial community structure during oral fluoropyrimidine treatment across multiple patient cohorts, in mouse small and large intestinal contents, and in patient-derived ex vivo communities were revealed by 16S rRNA gene sequencing. Metagenomic sequencing revealed marked shifts in pyrimidine-related gene abundance during oral fluoropyrimidine treatment, including enrichment of the preTA operon, which was sufficient for the inactivation of active metabolite 5-fluorouracil (5-FU). preTA+ bacteria depleted 5-FU in gut microbiota grown ex vivo and in the mouse distal gut. Germ-free and antibiotic-treated mice experienced increased fluoropyrimidine toxicity, which was rescued by colonization with the mouse gut microbiota, preTA+ Escherichia coli, or preTA-high stool from patients with CRC. Last, preTA abundance was negatively associated with fluoropyrimidine toxicity in patients. Together, these data support a causal, clinically relevant interaction between a human gut bacterial operon and the dose-limiting side effects of cancer treatment. Our approach may be generalizable to other drugs, including cancer immunotherapies, and provides valuable insights into host-microbiome interactions in the context of disease.

Animals↗