Search

The spread of disease by enteric pathogens associated with fecal contamination is a major concern for the management of urban watersheds. So far, the relative contribution of natural and anthropogenic sources to fecal pollution in managed tropical watersheds remains poorly evaluated. In this study, the microbiomes of water samples collected from managed watersheds in Singapore were elicited using the PhyloChip, a dense 16S rRNA gene-based DNA microarray, and fecal impairment was inferred using a machine-learning classification algorithm (SourceTracker). The predicted contribution of wildlife fecal sources to environmental samples was generally negligible (< 0.01 ± 0.01), indicating a low likelihood of fecal impairment from natural sources. However, sewage showed considerably higher contribution (0.09 ± 0.05) to microbial communities in a subset of watershed samples from canals and rivers, suggesting persistent impairment of certain areas by anthropogenic activity although being managed. Interestingly, the contribution of sewage microbial communities showed decreasing trends from canals/rivers to the connected reservoirs, indicating meaningful auto-mitigation of fecal pollution in canals and rivers. Notably, exclusion of locally derived fecal samples and source categories from the training data set impaired the predictive performance of the classification algorithm despite a high degree of similarity in the phylogenetic composition of microbiomes in biologically similar but geographically distinct sources. [Display omitted] • Inclusion of geographically relevant source samples improved category predictions. • A representative background source category improved SourceTracker reliability. • Contributions of human-related sources correlated with Bacteroides HF183 abundance. • Canals showed greater contributions from human-related fecal sources than reservoirs. [ABSTRACT FROM AUTHOR]

Harmful algal blooms (HAB) occur worldwide and cause health problems and economic damage to fisheries and tourism. Monitoring for toxic algae is therefore essential but is based primarily on light microscopy, which is time consuming and can be limited by insufficient morphological characters such that more time is needed to examine critical features with electron microscopy. Monitoring with molecular tools is done in only a few places world-wide. EU FP7 MIDTAL (Microarray Detection of Toxic Algae) used SSU and LSU rRNA genes as targets on microarrays to identify toxic species. In order to comply with current monitoring requirements to report cell numbers as the relevant threshold measurement to trigger closure of fisheries, it was necessary to calibrate our microarray to convert the hybridisation signal obtained to cell numbers. Calibration curves for two species of Pseudo-nitzschia for use with the MIDTAL microarray are presented to obtain cell numbers following hybridisation. It complements work presented by Barra et al. (2012b. Environ. Sci. Pollut. Res. doi: 10.1007/s11356-012-1330-1v) for two other Pseudo-nitzschia spp., Dittami and Edvardsen (2012a. J. Phycol. 48, 1050) for Pseudochatonella, Blanco et al. (2013. Harmful Algae 24, 80) for Heterosigma, McCoy et al. (2013. FEMS. doi: 10.1111/1574-6941.12277) for Prymnesium spp., Karlodinium veneficum, and cf. Chatonella spp. and Taylor et al. (2014. Harmful Algae, in press) for Alexandrium. [ABSTRACT FROM AUTHOR]

The dinoflagellate genus Alexandrium contains several toxin producing species and strains, which can cause major economic losses to the shell fish industry. It is therefore important to be able to detect these toxin producers and also distinguish toxic strains from some of the morphologically identical non-toxic strains. To facilitate this DNA probes to be used in a microarray format were designed in silico or developed from existing published probes. These probes targeted either the 18S or 28S ribosomal ribonucleic acid (rRNA) gene in Alexandrium tamarense Group I, Group III and Group IV, Alexandrium ostenfeldii and Alexandrium minutum. Three strains of A. tamarense Group I, A. tamarense Group III, A. minutum and two strains of A. ostenfeldii were grown at optimal conditions and transferred into new environmental conditions changing either the light intensity, salinity, temperature or nutrient concentrations, to check if any of these environmental conditions induced changes in the cellular ribonucleic acid (RNA) concentration or growth rate. The aim of this experiment was the calibration of several species-specific probes for the quantification of the toxic Alexandrium strains. Growth rates were highly variable but only elevated or lowered salinity significantly lowered growth rate for A. tamarense Group I and Group III; differences in RNA content were not significant for the majority of the treatments. Only light intensity seemed to affect significantly the RNA content in A. tamarense Group I and Group III, but this was still within the same range as for the other treatments meaning that a back calibration from RNA to cell numbers was possible. The designed probes allow the production of quantitative information for Alexandrium species for the microarray chip. [ABSTRACT FROM AUTHOR]

Abstract: Molecular microbial community analyses provide information on thousands of microorganisms simultaneously, and integrate biotic and abiotic perturbations caused by fecal contamination entering water bodies. A few studies have explored community methods as emerging approaches for microbial source tracking (MST), however, an evaluation of the current state of this approach is lacking. Here, we utilized three types of community-based methods with 64 blind, single- or dual-source, challenge samples generated from 12 sources, including: humans (feces), sewage, septage, dogs, pigs, deer, horses, cows, chickens, gulls, pigeons, and geese. Each source was a composite from multiple donors from four representative geographical regions in California. Methods evaluated included terminal restriction fragment polymorphism (TRFLP), phylogenetic microarray (PhyloChip), and next generation (Illumina) sequencing. These methods correctly identified dominant (or sole) sources in over 90% of the challenge samples, and exhibited excellent specificity regardless of source, rarely detecting a source that was not present in the challenge sample. Sensitivity, however, varied with source and community analysis method. All three methods distinguished septage from human feces and sewage, and identified deer and horse with 100% sensitivity and 100% specificity. Method performance improved if the composition of blind dual-source reference samples were defined by DNA contribution of each single source within the mixture, instead of by Enterococcus colony forming units. Data analysis approach also influenced method performance, indicating the need to standardize data interpretation. Overall, results of this study indicate that community analysis methods hold great promise as they may be used to identify any source, and they are particularly useful for sources that currently do not have, and may never have, a source-specific single marker gene. [Copyright &y& Elsevier]

Abstract: The biogeochemical processes that occur in marine sediments on continental margins are complex; however, from one perspective they can be considered with respect to three geochemical zones based on the presence and form of methane: sulfate–methane transition (SMTZ), gas hydrate stability zone (GHSZ), and free gas zone (FGZ). These geochemical zones may harbor distinct microbial communities that are important in biogeochemical carbon cycles. The objective of this study was to describe the microbial communities in sediments from the SMTZ, GHSZ, and FGZ using molecular ecology methods (i.e. PhyloChip microarray analysis and terminal restriction fragment length polymorphism (T-RFLP)) and examining the results in the context of non-biological parameters in the sediments. Non-metric multidimensional scaling and multi-response permutation procedures were used to determine whether microbial community compositions were significantly different in the three geochemical zones and to correlate samples with abiotic characteristics of the sediments. This analysis indicated that microbial communities from all three zones were distinct from one another and that variables such as sulfate concentration, hydrate saturation of the nearest gas hydrate layer, and depth (or unmeasured variables associated with depth e.g. temperature, pressure) were correlated to differences between the three zones. The archaeal anaerobic methanotrophs typically attributed to performing anaerobic oxidation of methane were not detected in the SMTZ; however, the marine benthic group-B, which is often found in SMTZ, was detected. Within the GHSZ, samples that were typically closer to layers that contained higher hydrate saturation had indicator sequences related to Vibrio-type taxa. These results suggest that the biogeographic patterns of microbial communities in marine sediments are distinct based on geochemical zones defined by methane. [Copyright &y& Elsevier]

Highlights: [•] Two anaerobic microbial consortia were established to depolymerize lignin. [•] The consortia could produce biomethane during lignin depolymerization. [•] The consortia could increase the hydrolytic efficiency of lignocellulosic biomass (e.g. OPEFB). [ABSTRACT FROM AUTHOR]

Abstract: Globally, tussock-based grasslands are being modified to increase productive capacity. The impacts of cultivation and over-sowing with exotic grass and legumes on soil microbiology were assessed at four sites in New Zealand which differed in soil type, climate and vegetation. Primary alteration of the soil physicochemical status occurred with land use change. This was driven by addition of mineral fertiliser and alteration of pH. Genes associated with several biogeochemical cycles (GeoChip data) were impacted by land-use but not sampling location. A number of functional gene families associated with biogeochemical cycling of C, N and S were present in greater relative abundance in the undisturbed soils. Similarly, soil bacterial (PhyloChip) and fungal (TRFLP) communities were strongly influenced by land-use change, but unaffected by sampling location. Alteration of land-use increased the relative abundance of Firmicutes, Actinobacteria and OD1 phyla, but many of the less-common phyla, such as Verrucomicrobia and Dictyoglomi decreased in abundance; these phyla may be important in internal soil nutrient cycling processes. This work provides evidence that tussock grassland soils are strongly dependent on microbially-mediated nutrient cycling, and these processes are highly-sensitive to exogenous nutrient inputs and/or alteration of pH. De-coupling of processes following addition of fertilisers or removal of organic matter (grazing) may make these improved grassland systems more susceptible to nutrient leakage. This has important implications for environmental quality. [Copyright &y& Elsevier]

Abstract: Soil microbial diversity is vast, and we lack even basic understanding of how this diversity is distributed ecologically. Using pyrosequencing and microarray methods, we quantified the structure of bacterial communities in two contrasting soils underlying Bornean rain forest (clay and sandy loam) that differ markedly in soil properties, aboveground tree flora, and leaf litter decomposition rates. We found significant soil-related taxonomic and phylogenetic differences between communities that, due to their proximity, are independent of climate. Bacterial communities showed distinct compositional and taxon-abundance distributions that were significantly correlated with the structure of the overlying tree community. Richness of bacteria was greater in the more resource-rich clay soil. Phylogenetic community analyses suggested that environmental filtering may be an important mechanism of community assembly in clay, compared to niche-competition in sandy loam. The Acidobacteria were the most abundant group in clay, but the Proteobacteria dominated in sandy loam. Of the ten most abundant classes, the Actinobacteria, Betaproteobacteria, Clostridia, Bacilli, and Gammaproteobacteria were more abundant in sandy loam than clay. Our study, which is the first to quantify edaphic variation in bacterial communities using high-throughput methods in soils underlying one of the most tree species rich forests on Earth, indicates an important role of plant–soil feedbacks linking the community structure of the trees and the underlying soil microbiome. We suggest the biochemical composition of carbon and nutrient resources in plant litter and soil pH and oxygen availability as important determinants of the distribution of bacterial diversity. [Copyright &y& Elsevier]

Background & Aims: The intestinal microbiomes of healthy children and pediatric patients with irritable bowel syndrome (IBS) are not well defined. Studies in adults have indicated that the gastrointestinal microbiota could be involved in IBS. Methods: We analyzed 71 samples from 22 children with IBS (pediatric Rome III criteria) and 22 healthy children, ages 7–12 years, by 16S ribosomal RNA gene sequencing, with an average of 54,287 reads/stool sample (average 454 read length = 503 bases). Data were analyzed using phylogenetic-based clustering (Unifrac), or an operational taxonomic unit (OTU) approach using a supervised machine learning tool (randomForest). Most samples were also hybridized to a microarray that can detect 8741 bacterial taxa (16S rRNA PhyloChip). Results: Microbiomes associated with pediatric IBS were characterized by a significantly greater percentage of the class γ-proteobacteria (0.07% vs 0.89% of total bacteria, respectively; P < .05); 1 prominent component of this group was Haemophilus parainfluenzae. Differences highlighted by 454 sequencing were confirmed by high-resolution PhyloChip analysis. Using supervised learning techniques, we were able to classify different subtypes of IBS with a success rate of 98.5%, using limited sets of discriminant bacterial species. A novel Ruminococcus-like microbe was associated with IBS, indicating the potential utility of microbe discovery for gastrointestinal disorders. A greater frequency of pain correlated with an increased abundance of several bacterial taxa from the genus Alistipes. Conclusions: Using16S metagenomics by PhyloChip DNA hybridization and deep 454 pyrosequencing, we associated specific microbiome signatures with pediatric IBS. These findings indicate the important association between gastrointestinal microbes and IBS in children; these approaches might be used in diagnosis of functional bowel disorders in pediatric patients. [ABSTRACT FROM AUTHOR]

Abstract: Sludge reduction is one of the major challenges in biological wastewater treatment. One approach is to increase the sludge degradation yield together with the biodegradation kinetics. Among the various sludge pretreatment strategies proposed, thermal pretreatment at around 65 °C was described as promising. The enhancement in the biodegradation activity due to the selection of thermophilic hydrolytic bacteria was proposed, but further experiments are needed to demonstrate the specific role of these bacteria. In this study, concentrated activated sludge grown at 20 °C was subjected to thermal treatment at 65 °C for different periods. The originality of the work relied on a polyphasic approach based on the correlation between kinetics (chemical oxygen demand, COD; mixed liquor suspended solids, MLSS), bacterial activity (respirometry) and bacterial community structure (phylochip monitoring) in order to characterize the mechanisms involved in the thermal reduction of sludge. The bacterial activity in the aeration basin decreased to a very low level when recycling sludge was treated at 65 °C from 13 to 60 h, but then, started to increase after 60 h. In parallel to these fluctuations in activity, a drastic shift occurred in the bacterial community structure with the selection of thermophilic bacteria (mainly related to genera Paenibacillus and Bacillus), which are known for their specific hydrolases. [ABSTRACT FROM AUTHOR]

Abstract: The arrival of chemicals in a soil or groundwater ecosystem could upset the natural balance of the microbial community. Since soil microorganisms are the first to be exposed to the chemicals released into the soil environment, we evaluated the use of a phylogenetic microarray as a bio-indicator of community perturbations due to the exposure to trichloroethylene (TCE). The phylogenetic microarray, which measures the presence of different members of the soil community, was used to evaluate unpolluted soils exposed to TCE as well as to samples from historically TCE polluted sites. We were able to determine an apparent threshold at which the microbial community structure was significantly affected (about 1ppm). In addition, the members of the microbial community most affected were identified. This approach could be useful for assessing environmental impact of chemicals on the biosphere as well as important members of the microbial community involved in TCE degradation. [Copyright &y& Elsevier]

Abstract: We designed an oligonucleotide microarray using probe sequences based upon a phylogenetic analysis of 16S rRNA genes recovered from members of the bacterial division Acidobacteria. A total of 42,194 oligonucleotide probes targeting members of the Acidobacteria division at multiple phylogenetic levels were included on a high-density microarray. Positive control hybridizations revealed a linear relationship between hybridization signal and template concentration, and a substantial decrease in non-specific hybridization was achieved through the addition of 2.5 M betaine to the hybridization buffer. A mean hybridization signal value was calculated for each Acidobacteria lineage, with the resultant lineage-specific hybridization data revealing strong predictive value for the positive control hybridizations. The Acidobacteria phylochip was then used to evaluate Acidobacteria rRNA genes from a Wisconsin soil and within a soil clay fraction. The Acidobacteria hybridization profile revealed the predominance of Acidobacteria subdivisions four and six, and also suggested a decrease in the abundance of subdivision six relative to subdivision four in the soil clay fraction. The change in relative abundance of these subdivisions in a soil clay fraction was supported by data from quantitative PCR. These results support the utility of a phylogenetic microarray in revealing changes in microbial population-level distributions in a complex soil microbial assemblage. [Copyright &y& Elsevier]

Abstract: A combination of culture-dependent and independent approaches was employed to identify the microbial community structure in a Greek solar saltern. A total of 219 and 132 isolates belonging, respectively, to Bacteria and Archaea, were recovered. All bacterial isolates were phylogenetically related to 43 members of Actinobacteria, Firmicutes and γ-Proteobacteria. The archaeal isolates were placed within the Halobacteriaceae. At least four groups of isolates represented novel species among the Bacteria. High bacterial diversity, consisting of 417 subfamilies, was revealed using a high-density oligonucleotide microarray (PhyloChip). At the four stages of saltern operation analyzed, the archaeal community consisted of both Crenarchaeota and Euryarchaeota, except for the sediment where Crenarchaeota were not detected. The bacterial community in sediment consisted mainly of γ-Proteobacteria and Actinobacteria, while, in hypersaline water, it was restricted to a few representatives of Bacteria. Members of α-Proteobacteria were the main constituents in saturated brine and crude salt, followed by γ-Proteobacteria, Actinobacteria and Firmicutes. A large Bacteroidetes and Verrucomicrobia diversity was identified in saturated brine, while δ-Proteobacteria and Cloroflexi were abundant in crude salt. Significant changes in the microbial community structure were detected during a short time period, denoting a rapidly adaptive dynamic ecosystem and viable diversity. Prokaryotic members reported for the first time in solar salterns were identified. [Copyright &y& Elsevier]

Pharmaceutical active compounds (PhACs) are molecules used for the treatment of physiological disorders that can generate environmental imbalances when discharged into wastewater. In this work, the use of Penicillium oxalicum XD-3.1 as bioremediation tool for the elimination of PhACs in non-sterile real hospital wastewater in batch bench scale bioreactor and its impact on microbial community was evaluated. A quantitative analysis using UHPLC-QToF was performed under three different conditions to determine the elimination of the different (PhACs) by this ascomycetes fungus. On the other hand, the changes on microbial community over time were analysed using Illumina MiSeq sequencing platform and a phylogenetic microarray (PhyloChip). P. oxalicum XD-3.1 was able to reduce the concentration of the majority of the analysed PhACs in 24 h such as ketoprofen, naproxen and paracetamol. Moreover, P. oxalicum inhibited the native fungal populations present in the wastewater, including opportunistic human pathogens such as Mycosphaerella and Drechslera, as well as some bacterial human pathogens belonging to Clostridiaceae and Brucellaceae families. However, important degraders such as Pseudomonadaceae bacteria groups remained in the system, indicating a possible natural consortium formation between fungi and bacteria. These results indicate the possibility to use this system for the removal of PhACs in real applications. Image 1 • P. oxalicum XD-3.1 was used as bioaugmented treatment in a fluidised batch bioreactor. • The fungus removed diclofenac, paracetamol and ketoprofen in 24 h. • Fungal populations related with human diseases decreased after bioaugmentation. • Bacterial biodiversity decreased during the treatment. • PhyloChip showed an accurate technology to detect rare populations. [ABSTRACT FROM AUTHOR]

• The European Project μAqua (FP7-KBBE-2010-4, 265409) was conceived as a project to make novel tools for the early and sensitive detection of freshwater-borne pathogens and cyanobacterial toxins. • The novel tools developed in μAqua were a phylochip (microarray), which identifies the presence of freshwater pathogenic targets using RNA barcodes and a RT-PCR microarray to amplify the mRNA captured by the barcodes for genes involved in cyanobacterial toxin synthesis. • We present results for three sites in along the Mediterranean coast of south-east France ranging from fresh to brackish to full strength seawater to test the applicability of the probes across the marine/freshwater boundary and infer the distribution of these pathogens across these boundaries. • The results for the cyanobacteria and Protozoa are reported here. • As freshwaters flow into marine systems, these pathogens and their toxins flow into a different environmental habitat. • Nevertheless these pathogens and toxins can still cause health problems in the marine environment and this paper shows the application of a phylochip across these different habitats. Monitoring drinking water quality is an important public health issue and pathogenic organisms present a particularly serious health hazard in freshwater bodies. However, many pathogenic bacteria, including cyanobacteria, and pathogenic protozoa can be swept into coastal lagoons and into near-shore marine environments where they continue to grow and pose a health threat to marine mammals and invertebrates. In this study, we tested the suitability of a phylochip (microarray for species detection) developed for freshwater pathogenic organisms to be applied to samples taken across a marine/freshwater interface at monthly intervals for two years. Toxic cyanobacteria and pathogenic protozoa were more numerous in a coastal lagoon than at the freshwater or marine site, indicating that this microarray can be used to detect the presence of these pathogens across a marine/freshwater interface and thus the potential for toxicity to occur within the entire watershed. [ABSTRACT FROM AUTHOR]

Background: Improvement in lung function after macrolide antibiotic therapy has been attributed to reduction in bronchial infection by specific bacteria. However, the airway might be populated by a more diverse microbiota, and clinical features of asthma might be associated with characteristics of the airway microbiota present. Objective: We sought to determine whether relationships exist between the composition of the airway bacterial microbiota and clinical features of asthma using culture-independent tools capable of detecting the presence and relative abundance of most known bacteria. Methods: In this pilot study bronchial epithelial brushings were collected from 65 adults with suboptimally controlled asthma participating in a multicenter study of the effects of clarithromycin on asthma control and 10 healthy control subjects. A combination of high-density 16S ribosomal RNA microarray and parallel clone library-sequencing analysis was used to profile the microbiota and examine relationships with clinical measurements. Results: Compared with control subjects, 16S ribosomal RNA amplicon concentrations (a proxy for bacterial burden) and bacterial diversity were significantly higher among asthmatic patients. In multivariate analyses airway microbiota composition and diversity were significantly correlated with bronchial hyperresponsiveness. Specifically, the relative abundance of particular phylotypes, including members of the Comamonadaceae, Sphingomonadaceae, Oxalobacteraceae, and other bacterial families were highly correlated with the degree of bronchial hyperresponsiveness. Conclusion: The composition of bronchial airway microbiota is associated with the degree of bronchial hyperresponsiveness among patients with suboptimally controlled asthma. These findings support the need for further functional studies to examine the potential contribution of members of the airway microbiota in asthma pathogenesis. [ABSTRACT FROM AUTHOR]