scholarly journals Application of a High-Density Oligonucleotide Microarray Approach To Study Bacterial Population Dynamics during Uranium Reduction and Reoxidation

2006 ◽  
Vol 72 (9) ◽  
pp. 6288-6298 ◽  
Author(s):  
Eoin L. Brodie ◽  
Todd Z. DeSantis ◽  
Dominique C. Joyner ◽  
Seung M. Baek ◽  
Joern T. Larsen ◽  
...  
Keyword(s):  
Microbial Community Composition ◽  
High Density ◽  
Rrna Gene ◽  
Bacterial Populations ◽  
Process Comparison ◽  
Reducing Conditions ◽  
Reducing Bacteria ◽  
Geothrix Fermentans ◽  
Metal Reducing Bacteria

ABSTRACT Reduction of soluble uranium U(VI) to less-soluble uranium U(IV) is a promising approach to minimize migration from contaminated aquifers. It is generally assumed that, under constant reducing conditions, U(IV) is stable and immobile; however, in a previous study, we documented reoxidation of U(IV) under continuous reducing conditions (Wan et al., Environ. Sci. Technol. 2005, 39:6162-6169). To determine if changes in microbial community composition were a factor in U(IV) reoxidation, we employed a high-density phylogenetic DNA microarray (16S microarray) containing 500,000 probes to monitor changes in bacterial populations during this remediation process. Comparison of the 16S microarray with clone libraries demonstrated successful detection and classification of most clone groups. Analysis of the most dynamic groups of 16S rRNA gene amplicons detected by the 16S microarray identified five clusters of bacterial subfamilies responding in a similar manner. This approach demonstrated that amplicons of known metal-reducing bacteria such as Geothrix fermentans (confirmed by quantitative PCR) and those within the Geobacteraceae were abundant during U(VI) reduction and did not decline during the U(IV) reoxidation phase. Significantly, it appears that the observed reoxidation of uranium under reducing conditions occurred despite elevated microbial activity and the consistent presence of metal-reducing bacteria. High-density phylogenetic microarrays constitute a powerful tool, enabling the detection and monitoring of a substantial portion of the microbial population in a routine, accurate, and reproducible manner.

scholarly journals Patterns in Wetland Microbial Community Composition and Functional Gene Repertoire Associated with Methane Emissions

mBio ◽  
2015 ◽  
Vol 6 (3) ◽  
Author(s):  
Shaomei He ◽  
Stephanie A. Malfatti ◽  
Jack W. McFarland ◽  
Frank E. Anderson ◽  
Amrita Pati ◽  
...  
Keyword(s):  
Microbial Community ◽  
Microbial Communities ◽  
Greenhouse Gas ◽  
Community Composition ◽  
Electron Acceptor ◽  
Methane Production ◽  
Microbial Community Composition ◽  
Restored Wetland ◽  
Reducing Bacteria ◽  
Metal Reducing Bacteria

ABSTRACTWetland restoration on peat islands previously drained for agriculture has potential to reverse land subsidence and sequester atmospheric carbon dioxide as peat accretes. However, the emission of methane could potentially offset the greenhouse gas benefits of captured carbon. As microbial communities play a key role in governing wetland greenhouse gas fluxes, we are interested in how microbial community composition and functions are associated with wetland hydrology, biogeochemistry, and methane emission, which is critical to modeling the microbial component in wetland methane fluxes and to managing restoration projects for maximal carbon sequestration. Here, we couple sequence-based methods with biogeochemical and greenhouse gas measurements to interrogate microbial communities from a pilot-scale restored wetland in the Sacramento-San Joaquin Delta of California, revealing considerable spatial heterogeneity even within this relatively small site. A number of microbial populations and functions showed strong correlations with electron acceptor availability and methane production; some also showed a preference for association with plant roots. Marker gene phylogenies revealed a diversity of major methane-producing and -consuming populations and suggested novel diversity within methanotrophs. Methanogenic archaea were observed in all samples, as were nitrate-, sulfate-, and metal-reducing bacteria, indicating that no single terminal electron acceptor was preferred despite differences in energetic favorability and suggesting spatial microheterogeneity and microniches. Notably, methanogens were negatively correlated with nitrate-, sulfate-, and metal-reducing bacteria and were most abundant at sampling sites with high peat accretion and low electron acceptor availability, where methane production was highest.IMPORTANCEWetlands are the largest nonanthropogenic source of atmospheric methane but also a key global carbon reservoir. Characterizing belowground microbial communities that mediate carbon cycling in wetlands is critical to accurately predicting their responses to changes in land management and climate. Here, we studied a restored wetland and revealed substantial spatial heterogeneity in biogeochemistry, methane production, and microbial communities, largely associated with the wetland hydraulic design. We observed patterns in microbial community composition and functions correlated with biogeochemistry and methane production, including diverse microorganisms involved in methane production and consumption. We found that methanogenesis gene abundance is inversely correlated with genes from pathways exploiting other electron acceptors, yet the ubiquitous presence of genes from all these pathways suggests that diverse electron acceptors contribute to the energetic balance of the ecosystem. These investigations represent an important step toward effective management of wetlands to reduce methane flux to the atmosphere and enhance belowground carbon storage.

Biogeochemical behaviour of plutonium during anoxic biostimulation of contaminated sediments

2012 ◽  
Vol 76 (3) ◽  
pp. 567-578 ◽  
Author(s):  
R. L. Kimber ◽  
C. Boothman ◽  
P. Purdie ◽  
F. R. Livens ◽  
J. R. Lloyd
Keyword(s):  
Contaminated Soils ◽  
Rrna Gene ◽  
Gene Profile ◽  
Environmental Behaviour ◽  
Reducing Conditions ◽  
Complex Chemistry ◽  
A Minor ◽  
Reducing Bacteria ◽  
The 16S Rrna Gene

AbstractUnderstanding the biogeochemical behaviour of actinides in the environment is essential for the longterm stewardship of radionuclide contaminated land. Plutonium is of particular concern due its high radiotoxicity, long half-life and complex chemistry, with these factors contributing to the limited literature available on its environmental behaviour. Here, we investigate the biogeochemistry of Pu in contaminated soil as microbial processes have the potential to mobilize Pu through numerous mechanisms including the reduction of Pu(IV) to the potentially more mobile Pu(III). After the addition of glucose to stimulate microbial activities, there was a substantial shift in the 16S rRNA gene profile of the extant microbial communities between days 0 and 44 with an increase in Clostridium species, known glucose fermenters which have been reported to facilitate the reduction of Pu(IV) to Pu(III). A minor increase in Pu mobility was observed at day 44, returning to initial levels by day 118. The negligible change in Pu mobility, despite the onset of reducing conditions and changing mineralogy, would suggest the Pu is highly refractory. This information is important for developing remediation options for Pu-contaminated soils, suggesting that managing legacy Pu in situ may be preferred to mobilization via the stimulation of metal-reducing bacteria.

scholarly journals Benchmarking laboratory processes to characterise low-biomass respiratory microbiota

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Raiza Hasrat ◽  
Jolanda Kool ◽  
Wouter A. A. de Steenhuijsen Piters ◽  
Mei Ling J. N. Chu ◽  
Sjoerd Kuiling ◽  
...  
Keyword(s):  
Microbial Community ◽  
Community Composition ◽  
Microbial Community Composition ◽  
Positive Control ◽  
Rrna Gene ◽  
Elution Buffer ◽  
Community Profile ◽  
Microbial Community Profile ◽  
Microbial Dna ◽  
Pcr Conditions

AbstractThe low biomass of respiratory samples makes it difficult to accurately characterise the microbial community composition. PCR conditions and contaminating microbial DNA can alter the biological profile. The objective of this study was to benchmark the currently available laboratory protocols to accurately analyse the microbial community of low biomass samples. To study the effect of PCR conditions on the microbial community profile, we amplified the 16S rRNA gene of respiratory samples using various bacterial loads and different number of PCR cycles. Libraries were purified by gel electrophoresis or AMPure XP and sequenced by V2 or V3 MiSeq reagent kits by Illumina sequencing. The positive control was diluted in different solvents. PCR conditions had no significant influence on the microbial community profile of low biomass samples. Purification methods and MiSeq reagent kits provided nearly similar microbiota profiles (paired Bray–Curtis dissimilarity median: 0.03 and 0.05, respectively). While profiles of positive controls were significantly influenced by the type of dilution solvent, the theoretical profile of the Zymo mock was most accurately analysed when the Zymo mock was diluted in elution buffer (difference compared to the theoretical Zymo mock: 21.6% for elution buffer, 29.2% for Milli-Q, and 79.6% for DNA/RNA shield). Microbiota profiles of DNA blanks formed a distinct cluster compared to low biomass samples, demonstrating that low biomass samples can accurately be distinguished from DNA blanks. In summary, to accurately characterise the microbial community composition we recommend 1. amplification of the obtained microbial DNA with 30 PCR cycles, 2. purifying amplicon pools by two consecutive AMPure XP steps and 3. sequence the pooled amplicons by V3 MiSeq reagent kit. The benchmarked standardized laboratory workflow presented here ensures comparability of results within and between low biomass microbiome studies.

scholarly journals 2,4-Dichlorophenoxyacetic acid degradation in methanogenic mixed cultures obtained from Brazilian Amazonian soil samples

2021 ◽  
Author(s):  
Gunther Brucha ◽  
Andrea Aldas-Vargas ◽  
Zacchariah Ross ◽  
Peng Peng ◽  
Siavash Atashgahi ◽  
...  
Keyword(s):  
Microbial Community Composition ◽  
High Mobility ◽  
16S Rrna Gene Sequencing ◽  
Dichlorophenoxyacetic Acid ◽  
Rrna Gene ◽  
Deep Soil ◽  
Anoxic Environments ◽  
Top Soil ◽  
Significant Enrichment ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract2,4-Dichlorophenoxyacetic acid (2,4-D) is the third most applied pesticide in Brazil to control broadleaf weeds in crop cultivation and pastures. Due to 2,4-D’s high mobility and long half-life under anoxic conditions, this herbicide has high probability for groundwater contamination. Bioremediation is an attractive solution for 2,4-D contaminated anoxic environments, but there is limited understanding of anaerobic 2,4-D biodegradation. In this study, methanogenic enrichment cultures were obtained from Amazonian top soil (0—40 cm) and deep soil (50 -80 cm below ground) that biotransform 2,4-D (5 µM) to 4-chlorophenol and phenol. When these cultures were transferred (10% v/v) to fresh medium containing 40 µM or 160 µM 2,4-D, the rate of 2,4-D degradation decreased, and biotransformation did not proceed beyond 4-chlorophenol and 2,4-dichlorophenol in the top and deep soil cultures, respectively. 16S rRNA gene sequencing and qPCR of a selection of microbes revealed no significant enrichment of known organohalide-respiring bacteria. Furthermore, a member of the genus Cryptanaerobacter was identified as possibly responsible for phenol conversion to benzoate in the top soil inoculated culture. Overall, these results demonstrate the effect of 2,4-D concentration on biodegradation and microbial community composition, which are both important factors when developing pesticide bioremediation technologies.

scholarly journals Structure of Microbial Communities When Complementary Effluents Are Anaerobically Digested

2021 ◽  
Vol 11 (3) ◽  
pp. 1293
Author(s):  
Ana Eusébio ◽  
André Neves ◽  
Isabel Paula Marques
Keyword(s):  
Anaerobic Digestion ◽  
Microbial Communities ◽  
Volatile Fatty Acids ◽  
Olive Mill Wastewater ◽  
Organic Load ◽  
Rrna Gene ◽  
Bacterial Populations ◽  
Next Generation Sequencing Ngs ◽  
Generation Sequencing ◽  
Olive Mill

Olive oil and pig productions are important industries in Portugal that generate large volumes of wastewater with high organic load and toxicity, raising environmental concerns. The principal objective of this study is to energetically valorize these organic effluents—piggery effluent and olive mill wastewater—through the anaerobic digestion to the biogas/methane production, by means of the effluent complementarity concept. Several mixtures of piggery effluent were tested, with an increasing percentage of olive mill wastewater. The best performance was obtained for samples of piggery effluent alone and in admixture with 30% of OMW, which provided the same volume of biogas (0.8 L, 70% CH4), 63/75% COD removal, and 434/489 L CH4/kg SVin, respectively. The validation of the process was assessed by molecular evaluation through Next Generation Sequencing (NGS) of the 16S rRNA gene. The structure of the microbial communities for both samples, throughout the anaerobic process, was characterized by the predominance of bacterial populations belonging to the phylum Firmicutes, mainly Clostridiales, with Bacteroidetes being the subdominant populations. Archaea populations belonging to the genus Methanosarcina became predominant throughout anaerobic digestion, confirming the formation of methane mainly from acetate, in line with the greatest removal of volatile fatty acids (VFAs) in these samples.

scholarly journals Do initial concentration and activated sludge seasonality affect pharmaceutical biotransformation rate constants?

2021 ◽  
Author(s):  
Tamara J. H. M. van Bergen ◽  
Ana B. Rios-Miguel ◽  
Tom M. Nolte ◽  
Ad M. J. Ragas ◽  
Rosalie van Zelm ◽  
...  
Keyword(s):  
Microbial Community ◽  
Activated Sludge ◽  
Community Composition ◽  
Rate Constants ◽  
Wastewater Treatment Plants ◽  
Microbial Community Composition ◽  
Amplicon Sequencing ◽  
Rrna Gene ◽  
Initial Concentration ◽  
Different Seasons

Abstract Pharmaceuticals find their way to the aquatic environment via wastewater treatment plants (WWTPs). Biotransformation plays an important role in mitigating environmental risks; however, a mechanistic understanding of involved processes is limited. The aim of this study was to evaluate potential relationships between first-order biotransformation rate constants (kb) of nine pharmaceuticals and initial concentration of the selected compounds, and sampling season of the used activated sludge inocula. Four-day bottle experiments were performed with activated sludge from WWTP Groesbeek (The Netherlands) of two different seasons, summer and winter, spiked with two environmentally relevant concentrations (3 and 30 nM) of pharmaceuticals. Concentrations of the compounds were measured by LC–MS/MS, microbial community composition was assessed by 16S rRNA gene amplicon sequencing, and kb values were calculated. The biodegradable pharmaceuticals were acetaminophen, metformin, metoprolol, terbutaline, and phenazone (ranked from high to low biotransformation rates). Carbamazepine, diatrizoic acid, diclofenac, and fluoxetine were not converted. Summer and winter inocula did not show significant differences in microbial community composition, but resulted in a slightly different kb for some pharmaceuticals. Likely microbial activity was responsible instead of community composition. In the same inoculum, different kb values were measured, depending on initial concentration. In general, biodegradable compounds had a higher kb when the initial concentration was higher. This demonstrates that Michealis-Menten kinetic theory has shortcomings for some pharmaceuticals at low, environmentally relevant concentrations and that the pharmaceutical concentration should be taken into account when measuring the kb in order to reliably predict the fate of pharmaceuticals in the WWTP. Key points • Biotransformation and sorption of pharmaceuticals were assessed in activated sludge. • Higher initial concentrations resulted in higher biotransformation rate constants for biodegradable pharmaceuticals. • Summer and winter inocula produced slightly different biotransformation rate constants although microbial community composition did not significantly change. Graphical abstract

scholarly journals Lachnospiraceae and Bacteroidales Alternative Fecal Indicators Reveal Chronic Human Sewage Contamination in an Urban Harbor

2011 ◽  
Vol 77 (19) ◽  
pp. 6972-6981 ◽  
Author(s):  
Ryan J. Newton ◽  
Jessica L. VandeWalle ◽  
Mark A. Borchardt ◽  
Marc H. Gorelick ◽  
Sandra L. McLellan
Keyword(s):  
Sequence Analysis ◽  
Genetic Marker ◽  
Microbial Community Composition ◽  
Fold Increase ◽  
Fecal Pollution ◽  
Plate Count ◽  
Rrna Gene ◽  
Fecal Indicators ◽  
Fecal Indicator ◽  
Content Type

ABSTRACTThe complexity of fecal microbial communities and overlap among human and other animal sources have made it difficult to identify source-specific fecal indicator bacteria. However, the advent of next-generation sequencing technologies now provides increased sequencing power to resolve microbial community composition within and among environments. These data can be mined for information on source-specific phylotypes and/or assemblages of phylotypes (i.e., microbial signatures). We report the development of a new genetic marker for human fecal contamination identified through microbial pyrotag sequence analysis of the V6 region of the 16S rRNA gene. Sequence analysis of 37 sewage samples and comparison with database sequences revealed a human-associated phylotype within theLachnospiraceaefamily, which was closely related to the genusBlautia. This phylotype, termed Lachno2, was on average the second most abundant fecal bacterial phylotype in sewage influent samples from Milwaukee, WI. We developed a quantitative PCR (qPCR) assay for Lachno2 and used it along with the qPCR-based assays for humanBacteroidales(based on the HF183 genetic marker), totalBacteroidalesspp., and enterococci and the conventionalEscherichia coliand enterococci plate count assays to examine the prevalence of fecal and human fecal pollution in Milwaukee's harbor. Both the conventional fecal indicators and the human-associated indicators revealed chronic fecal pollution in the harbor, with significant increases following heavy rain events and combined sewer overflows. The two human-associated genetic marker abundances were tightly correlated in the harbor, a strong indication they target the same source (i.e., human sewage). Human adenoviruses were routinely detected under all conditions in the harbor, and the probability of their occurrence increased by 154% for every 10-fold increase in the human indicator concentration. Both Lachno2 and humanBacteroidalesincreased specificity to detect sewage compared to general indicators, and the relationship to a human pathogen group suggests that the use of these alternative indicators will improve assessments for human health risks in urban waters.

scholarly journals Biogeochemical Modelling of Uranium Immobilization and Aquifer Remediation Strategies Near NCCP Sludge Storage Facilities

2021 ◽  
Vol 11 (6) ◽  
pp. 2875
Author(s):  
A.V. Safonov ◽  
A.E. Boguslavsky ◽  
O.L. Gaskova ◽  
K.A. Boldyrev ◽  
O.S. Shvartseva ◽  
...  
Keyword(s):  
Sandy Loam ◽  
Influential Factor ◽  
Rrna Gene ◽  
16S Rrna Gene Analysis ◽  
Long Term Stability ◽  
Microbial Biofilms ◽  
Remediation Strategies ◽  
Reducing Bacteria ◽  
Nitrate Reducing Bacteria ◽  
Storage Facilities

Nitrate is a substance which influences the prevailing redox conditions in groundwater, and in turn the behaviour of U. The study of groundwater in an area with low-level radioactive sludge storage facilities has shown their contamination with sulphate and nitrate anions, uranium, and some associated metals. The uranyl ion content in the most contaminated NO3–Cl–SO4–Na borehole is 2000 times higher (1.58 mg/L) than that in the background water. At the same time, assessment of the main physiological groups of microorganisms showed a maximum number of denitrifying and sulphate-reducing bacteria (e.g., Sulfurimonas) in the water from the same borehole. Biogenic factors of radionuclide immobilization on sandy rocks of upper aquifers have been experimentally investigated. Different reduction rates of NO3-, SO42-, Fe(III) and U(VI) with stimulated microbial activity were dependent on the pollution degree. Moreover, 16S rRNA gene analysis of the microbial community after whey addition revealed a significant decrease in microbial diversity and the activation of nonspecific nitrate-reducing bacteria (genera Rhodococcus and Rhodobacter). The second influential factor can be identified as the formation of microbial biofilms on the sandy loam samples, which has a positive effect on U sorption (an increase in Kd value is up to 35%). As PHREEQC physicochemical modelling numerically confirmed, the third most influential factor that drives U mobility is the biogenic-mediated formation of a sulphide redox buffer. This study brings important information, which helps to assess the long-term stability of U in the environment of radioactive sludge storage facilities.

scholarly journals Competitive Reduction of Pertechnetate (99TcO4−) by Dissimilatory Metal Reducing Bacteria and Biogenic Fe(II)

2011 ◽  
Vol 45 (3) ◽  
pp. 951-957 ◽  
Author(s):  
Andrew E. Plymale ◽  
James K. Fredrickson ◽  
John M. Zachara ◽  
Alice C. Dohnalkova ◽  
Steve M. Heald ◽  
...  
Keyword(s):  
Competitive Reduction ◽  
Reducing Bacteria ◽  
Metal Reducing Bacteria

scholarly journals A new pipeline for structural characterization and classification of RNA-Seq microbiome data

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Sebastian Racedo ◽  
Ivan Portnoy ◽  
Jorge I. Vélez ◽  
Homero San-Juan-Vergara ◽  
Marco Sanjuan ◽  
...  
Keyword(s):  
Classification Accuracy ◽  
High Throughput Sequencing ◽  
Compositional Data ◽  
Graphical Representation ◽  
Synthetic Data ◽  
Gene Sequencing ◽  
Rrna Gene ◽  
Interaction Patterns ◽  
Simulation Experiments

Abstract Background High-throughput sequencing enables the analysis of the composition of numerous biological systems, such as microbial communities. The identification of dependencies within these systems requires the analysis and assimilation of the underlying interaction patterns between all the variables that make up that system. However, this task poses a challenge when considering the compositional nature of the data coming from DNA-sequencing experiments because traditional interaction metrics (e.g., correlation) produce unreliable results when analyzing relative fractions instead of absolute abundances. The compositionality-associated challenges extend to the classification task, as it usually involves the characterization of the interactions between the principal descriptive variables of the datasets. The classification of new samples/patients into binary categories corresponding to dissimilar biological settings or phenotypes (e.g., control and cases) could help researchers in the development of treatments/drugs. Results Here, we develop and exemplify a new approach, applicable to compositional data, for the classification of new samples into two groups with different biological settings. We propose a new metric to characterize and quantify the overall correlation structure deviation between these groups and a technique for dimensionality reduction to facilitate graphical representation. We conduct simulation experiments with synthetic data to assess the proposed method’s classification accuracy. Moreover, we illustrate the performance of the proposed approach using Operational Taxonomic Unit (OTU) count tables obtained through 16S rRNA gene sequencing data from two microbiota experiments. Also, compare our method’s performance with that of two state-of-the-art methods. Conclusions Simulation experiments show that our method achieves a classification accuracy equal to or greater than 98% when using synthetic data. Finally, our method outperforms the other classification methods with real datasets from gene sequencing experiments.

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