Spatial scale structure soil bacterial communities across an Arctic landscape.

Bacterial community composition is largely influenced by environmental factors, and this applies to the Arctic region. However, little is known about the role of spatial factors in structuring such communities. In this study, we evaluated the influence of spatial scale on bacterial community structure across an Arctic landscape. Our results showed that spatial factors accounted for approximately 10% of the variation at the landscape scale, equivalent to observations across the whole Arctic region, suggesting that while the role and magnitude of other processes involved in community structure may vary, the role of dispersal may be stable globally in the region. We assessed dispersal limitation by identifying the spatial autocorrelation distance, standing at approximately 60 m, which would be required in order to obtain fully independent samples and may inform future sampling strategies in the region. Finally, indicator taxa with strong statistical correlations with environment variables were identified. However, we showed that these strong taxa-environment associations may not always be reflected in the geographical distribution of these taxa.IMPORTANCE The significance of this study is threefold. It investigated the influence of spatial scale on the soil bacterial community composition across a typical Arctic landscape and demonstrated that conclusions reached when examining the influence of specific environmental variables on bacterial community composition are dependent upon the spatial scales over which they are investigated. This study identified a dispersal limitation (spatial autocorrelation) distance of approximately 60 m, required to obtain samples with fully independent bacterial communities, and therefore, should serve to inform future sampling strategies in the region and potentially elsewhere. The work also showed that strong taxa-environment statistical associations may not be reflected in the observed landscape distribution of the indicator taxa.

Meltwater export of prokaryotic cells from the Greenland ice sheet.

Microorganisms are flushed from the Greenland Ice Sheet (GrIS) where they may contribute towards the nutrient cycling and community compositions of downstream ecosystems. We investigate meltwater microbial assemblages as they exit the GrIS from a large outlet glacier, and as they enter a downstream river delta during the record melt year of 2012. Prokaryotic abundance, flux and community composition was studied, and factors affecting community structures were statistically considered. The mean concentration of cells exiting the ice sheet was 8.30 × 104 cells mL-1 and we estimate that ∼1.02 × 1021 cells were transported to the downstream fjord in 2012, equivalent to 30.95 Mg of carbon. Prokaryotic microbial assemblages were dominated by Proteobacteria, Bacteroidetes, and Actinobacteria. Cell concentrations and community compositions were stable throughout the sample period, and were statistically similar at both sample sites. Based on our observations, we argue that the subglacial environment is the primary source of the river-transported microbiota, and that cell export from the GrIS is dependent on discharge. We hypothesise that the release of subglacial microbiota to downstream ecosystems will increase as freshwater flux from the GrIS rises in a warming world.

Potential Activity of Subglacial Microbiota Transported to Anoxic River Delta Sediments.

The Watson River drains a portion of the SW Greenland ice sheet, transporting microbial communities from subglacial environments to a delta at the head of Søndre Strømfjord. This study investigates the potential activity and community shifts of glacial microbiota deposited and buried under layers of sediments within the river delta. A long-term (12-month) incubation experiment was established using Watson River delta sediment under anaerobic conditions, with and without CO2/H2 enrichment. Within CO2/H2-amended incubations, sulphate depletion and a shift in the microbial community to a 52% predominance of Desulfosporosinus meridiei by day 371 provides evidence for sulphate reduction. We found evidence of methanogenesis in CO2/H2-amended incubations within the first 5 months, with production rates of ~4 pmol g-1 d-1, which was likely performed by methanogenic Methanomicrobiales- and Methanosarcinales-related organisms. Later, a reduction in methane was observed to be paired with the depletion of sulphate, and we hypothesise that sulphate reduction out competed hydrogenotrophic methanogenesis. The structure and diversity of the original CO2/H2-amended incubation communities changed dramatically with a major shift in predominant community members and a decline in diversity and cell abundance. These results highlight the need for further investigations into the fate of subglacial microbiota within downstream environments.

Polymorphonuclear leukocytes restrict growth of Pseudomonas aeruginosa in the lungs of cystic fibrosis patients.

Cystic fibrosis (CF) patients have increased susceptibility to chronic lung infections by Pseudomonas aeruginosa, but the ecophysiology within the CF lung during infections is poorly understood. The aim of this study was to elucidate the in vivo growth physiology of P. aeruginosa within lungs of chronically infected CF patients. A novel, quantitative peptide nucleic acid (PNA) fluorescence in situ hybridization (PNA-FISH)-based method was used to estimate the in vivo growth rates of P. aeruginosa directly in lung tissue samples from CF patients and the growth rates of P. aeruginosa in infected lungs in a mouse model. The growth rate of P. aeruginosa within CF lungs did not correlate with the dimensions of bacterial aggregates but showed an inverse correlation to the concentration of polymorphonuclear leukocytes (PMNs) surrounding the bacteria. A growth-limiting effect on P. aeruginosa by PMNs was also observed in vitro, where this limitation was alleviated in the presence of the alternative electron acceptor nitrate. The finding that P. aeruginosa growth patterns correlate with the number of surrounding PMNs points to a bacteriostatic effect by PMNs via their strong O2 consumption, which slows the growth of P. aeruginosa in infected CF lungs. In support of this, the growth of P. aeruginosa was significantly higher in the respiratory airways than in the conducting airways of mice. These results indicate a complex host-pathogen interaction in chronic P. aeruginosa infection of the CF lung whereby PMNs slow the growth of the bacteria and render them less susceptible to antibiotic treatment while enabling them to persist by anaerobic respiration.

Bacterial diversity in snow on North Pole ice floes.

The microbial abundance and diversity in snow on ice floes at three sites near the North Pole was assessed using quantitative PCR and 454 pyrosequencing. Abundance of 16S rRNA genes in the samples ranged between 43 and 248 gene copies per millilitre of melted snow. A total of 291,331 sequences were obtained through 454 pyrosequencing of 16S rRNA genes, resulting in 984 OTUs at 97 % identity. Two sites were dominated by Cyanobacteria (72 and 61 %, respectively), including chloroplasts. The third site differed by consisting of 95 % Proteobacteria. Principal component analysis showed that the three sites clustered together when compared to the underlying environments of sea ice and ocean water. The Shannon indices ranged from 2.226 to 3.758, and the Chao1 indices showed species richness between 293 and 353 for the three samples. The relatively low abundances and diversity found in the samples indicate a lower rate of microbial input to this snow habitat compared to snow in the proximity of terrestrial and anthropogenic sources of microorganisms. The differences in species composition and diversity between the sites show that apparently similar snow habitats contain a large variation in biodiversity, although the differences were smaller than the differences to the underlying environment. The results support the idea that a globally distributed community exists in snow and that the global snow community can in part be attributed to microbial input from the atmosphere.

Ageing of atrazine in manure amended soils assessed by bioavailability to Pseudomonas sp. strain ADP.

Animal manure is applied to agricultural land in areas of high livestock production. In the present study, we evaluated ageing of atrazine in two topsoils with and without addition of manure and in one subsoil. Ageing was assessed as the bioavailability of atrazine to the atrazine mineralizing bacteria Pseudomonas sp. strain ADP. Throughout an ageing period of 90 days bioavailability was investigated at days 1, 10, 32, 60 and 90, where ~10(8) cells g(-1) of the ADP strain was inoculated to the (14)C-atrazine exposed soil and (14)CO2 was collected over 7 days as a measure of mineralized atrazine. Even though the bioavailable residue decreased in all of the three soils as time proceeded, we found that ageing occurred faster in the topsoils rich in organic carbon than in subsoil. For one topsoil rich in organic carbon content, Simmelkær, we observed a higher degree of ageing when treated with manure. Contrarily, sorption experiments showed less sorption to Simmelkær treated with manure than the untreated soil indicating that sorption processes are not the only mechanisms of ageing. The other topsoil low in organic carbon content, Ringe, showed no significant difference in ageing between the manure-treated and untreated soil. The present study illustrates that not simply the organic carbon content influences adsorption and ageing of atrazine in soil but the origin and composition of organic matter plays an important role.

Adhesion of and to soil in runoff as influenced by polyacrylamide.

Polyacrylamide (PAM) is used in agriculture to reduce soil erosion and has been reported to reduce turbidity, nutrients, and pollutants in surface runoff water. The objective of this work was to determine the effect of PAM on the concentration of enteric bacteria in surface runoff by comparing four enteric bacteria representing phenotypically different motility and hydrophobicity from three soils. Results demonstrated that bacterial surface runoff was differentially influenced by the PAM treatment. Polyacrylamide treatment increased surface runoff for adhered and planktonic cells from a clay soil; significantly decreased surface runoff of adhered bacteria, while no difference was observed for planktonic bacteria from the sandy loam; and significantly decreased the surface runoff of planktonic cells, while no difference was observed for adhered bacteria from the clay loam. Comparing strains from a final water sample collected after 48 h showed a greater loss of while serovar Poona was almost not detected. Thus, (i) the PAM efficiency in reducing the concentration of enteric bacteria in surface runoff was influenced by soil type and (ii) variation in the loss of enteric bacteria highlights the importance of strain-specific properties that may not be captured with general fecal indicator bacteria.

Abrupt permafrost thaw triggers activity of copiotrophs and microbiome predators.

Permafrost soils store a substantial part of the global soil carbon and nitrogen. However, global warming causes abrupt erosion and gradual thaw, which make these stocks vulnerable to microbial decomposition into greenhouse gases. Here, we investigated the microbial response to abrupt in situ permafrost thaw. We sequenced the total RNA of a 1 m deep soil core consisting of up to 26 500-year-old permafrost material from an active abrupt erosion site. We analysed the microbial community in the active layer soil, the recently thawed, and the intact permafrost, and found maximum RNA:DNA ratios in recently thawed permafrost indicating a high microbial activity. In thawed permafrost, potentially copiotrophic Burkholderiales and Sphingobacteriales, but also microbiome predators dominated the community. Overall, both thaw-dependent and long-term soil properties significantly correlated with changes in community composition, as did microbiome predator abundance. Bacterial predators were dominated in shallower depths by Myxococcota, while protozoa, especially Cercozoa and Ciliophora, almost tripled in relative abundance in thawed layers. Our findings highlight the ecological importance of a diverse interkingdom and active microbial community highly abundant in abruptly thawing permafrost, as well as predation as potential biological control mechanism.

Microbial degradation of 2,4-dichlorophenoxyacetic acid on the Greenland ice sheet.

The Greenland ice sheet (GrIS) receives organic carbon (OC) of anthropogenic origin, including pesticides, from the atmosphere and/or local sources, and the fate of these compounds in the ice is currently unknown. The ability of supraglacial heterotrophic microbes to mineralize different types of OC is likely a significant factor determining the fate of anthropogenic OC on the ice sheet. Here we determine the potential of the microbial community from the surface of the GrIS to mineralize the widely used herbicide 2,4-dichlorophenoxyacetic acid (2,4-D). Surface ice cores were collected and incubated for up to 529 days in microcosms simulating in situ conditions. Mineralization of side chain- and ring-labeled [(14)C]2,4-D was measured in the samples, and quantitative PCR targeting the tfdA genes in total DNA extracted from the ice after the experiment was performed. We show that the supraglacial microbial community on the GrIS contains microbes that are capable of degrading 2,4-D and that they are likely present in very low numbers. They can mineralize 2,4-D at a rate of up to 1 nmol per m(2) per day, equivalent to ∼26 ng C m(-2) day(-1). Thus, the GrIS should not be considered a mere reservoir of all atmospheric contaminants, as it is likely that some deposited compounds will be removed from the system via biodegradation processes before their potential release due to the accelerated melting of the ice sheet.

Modeling of phenoxy acid herbicide mineralization and growth of microbial degraders in 15 soils monitored by quantitative real-time PCR of the functional tfdA gene.

Mineralization potentials, rates, and kinetics of the three phenoxy acid (PA) herbicides, 2,4-dichlorophenoxyacetic acid (2,4-D), 4-chloro-2-methylphenoxyacetic acid (MCPA), and 2-(4-chloro-2-methylphenoxy)propanoic acid (MCPP), were investigated and compared in 15 soils collected from five continents. The mineralization patterns were fitted by zero/linear or exponential growth forms of the three-half-order models and by logarithmic (log), first-order, or zero-order kinetic models. Prior and subsequent to the mineralization event, tfdA genes were quantified using real-time PCR to estimate the genetic potential for degrading PA in the soils. In 25 of the 45 mineralization scenarios, ∼60% mineralization was observed within 118 days. Elevated concentrations of tfdA in the range 1 × 10(5) to 5 × 10(7) gene copies g(-1) of soil were observed in soils where mineralization could be described by using growth-linked kinetic models. A clear trend was observed that the mineralization rates of the three PAs occurred in the order 2,4-D > MCPA > MCPP, and a correlation was observed between rapid mineralization and soils exposed to PA previously. Finally, for 2,4-D mineralization, all seven mineralization patterns which were best fitted by the exponential model yielded a higher tfdA gene potential after mineralization had occurred than the three mineralization patterns best fitted by the Lin model.

Microbial Community Changes in 26,500-Year-Old Thawing Permafrost.

Northern permafrost soils store more than half of the global soil carbon. Frozen for at least two consecutive years, but often for millennia, permafrost temperatures have increased drastically in the last decades. The resulting thermal erosion leads not only to gradual thaw, resulting in an increase of seasonally thawing soil thickness, but also to abrupt thaw events, such as sudden collapses of the soil surface. These could affect 20% of the permafrost zone and half of its organic carbon, increasing accessibility for deeper rooting vegetation and microbial decomposition into greenhouse gases. Knowledge gaps include the impact of permafrost thaw on the soil microfauna as well as key taxa to change the microbial mineralization of ancient permafrost carbon stocks during erosion. Here, we present the first sequencing study of an abrupt permafrost erosion microbiome in Northeast Greenland, where a thermal erosion gully collapsed in the summer of 2018, leading to the thawing of 26,500-year-old permafrost material. We investigated which soil parameters (pH, soil carbon content, age and moisture, organic and mineral horizons, and permafrost layers) most significantly drove changes of taxonomic diversity and the abundance of soil microorganisms in two consecutive years of intense erosion. Sequencing of the prokaryotic 16S rRNA and fungal ITS2 gene regions at finely scaled depth increments revealed decreasing alpha diversity with depth, soil age, and pH. The most significant drivers of variation were found in the soil age, horizons, and permafrost layer for prokaryotic and fungal beta diversity. Permafrost was mainly dominated by Proteobacteria and Firmicutes, with Polaromonas identified as the most abundant taxon. Thawed permafrost samples indicated increased abundance of several copiotrophic phyla, such as Bacteroidia, suggesting alterations of carbon utilization pathways within eroding permafrost.

Enhanced fermentative lactic acid production from source-sorted organic household waste: Focusing on low-pH microbial adaptation and bio-augmentation strategy.

Lactic acid (LA) production at low pH could significantly reduce the need for neutralizing agents, leading to reduction of operational costs. In the present study, LA production at acidic conditions was investigated using source-sorted organic household waste (SSOHW). Controlling the pH at low value (i.e. 5.0) and bio-augmenting with Pediococcus acidilactici led to a concentration of 39.3 ± 0.5 g-LA/L with a yield of 0.75 ± 0.02 g-LA/g-sugar. In contrast, secondary fermentation at higher pH level (i.e. 5.5 and 6.0) resulted in complete LA degradation. Subsequently, consecutive batch fermentations were conducted to adapt P. acidilactici to SSOHW and improve the LA production. Results showed that P. acidilactici could successively adapt in the SSOHW reaching a relative abundance above 2.8% at adaptation process. The added P. acidilactici ensured a high concentration of LA at three consecutive generations, achieving an increment above 18% compared to control test (abiotic augmentation). Moreover, adaptation processes (i.e. maintaining pH at 4.0 or stepwise decreasing the pH from 5.0 to 4.0) significantly improved LA concentration and productivity at the pH of 4.0. Overall, the results provide a promising method to reduce the LA production costs using residual resources.

Ex-situ biogas upgrading in thermophilic trickle bed reactors packed with micro-porous packing materials.

Two thermophilic trickle bed reactors (TBRs) were packed with different packing densities with polyurethane foam (PUF) and their performance under different retention times were evaluated during ex-situ biogas upgrading process. The results showed that the TBR more tightly packed i.e. containing more layers of PUF achieved higher H2 utilization efficiency (>99%) and thus, higher methane content (>95%) in the output gas. The tightly packed micro-porous PUF enhanced biofilm immobilization, gas-liquid mass transfer and biomethanation efficiency. Moreover, applying a continuous high-rate nutrient trickling could lead to liquid overflow resulting in formation of non-homogenous biofilm and severe deduction of biomethanation efficiency. High-throughput 16S rRNA gene sequencing revealed that the liquid media were predominated by hydrogenotrophic methanogens. Moreover, members of Peptococcaceae family and uncultured members of Clostridia class were identified as the most abundant species in the biofilm. The proliferation of hydrogenotrophic methanogens together with syntrophic bacteria showed that H2 addition resulted in altering the microbial community in biogas upgrading process.

Diversity and Structure of Bacterial Communities in Different Rhizocompartments (Rhizoplane, Rhizosphere, and Bulk) at Flag Leaf Emergence in Four Winter Wheat Varieties.

Understanding basic interactions at the plant-soil interphase is critical if we are to exploit natural microbial communities for improved crop resilience. We report here 16S amplicon sequencing data from 3 rhizocompartments of 4 wheat cultivars grown under controlled greenhouse conditions. We observed that rhizocompartments and cultivar affect the community composition.

Microbial Diversity in Four Rhizocompartments (Bulk Soil, Rhizosphere, Rhizoplane, and Endosphere) of Four Winter Wheat Varieties at the Fully Emerged Flag Leaf Growth Stage.

Community composition and recruitment are important elements of plant-microbe interactions and may provide insights for plant development and resilience. The results of 16S rRNA amplicon sequencing from four rhizocompartments for four wheat cultivars grown under controlled conditions and sampled after flag leaf emergence are provided. Data demonstrate differences in microbial communities according to rhizocompartment.

Ex-situ biogas upgrading in thermophilic up-flow reactors: The effect of different gas diffusers and gas retention times.

Four different types of ceramic gas distributors (Al2O3 of 1.2 µm and SiC of 0.5, 7 and 14 µm) were evaluated to increase biomethane formation during ex-situ biogas upgrading process. Each type of gas diffuser was tested independently at three different gas retention times of 10, 5 and 2.5 h, at thermophilic conditions. CH4 production rate increased by increasing input gas flow rate for all type of distributors, whereas CH4 concentration declined. Reactors equipped with SiC gas distributors effectively improved biomethane content fulfilling natural gas standards. Microbial analysis showed high abundance of hydrogenotrophic methanogens and proliferated syntrophic bacteria, i.e. syntrophic acetate oxidizers and homoacetogens, confirming the effect of H2 to alternate anaerobic digestion microbiome and enhance hydrogenotrophic methanogenesis. A detailed anaerobic bioconversion model was adapted to simulate the operation of the R1-R4 reactors. The model was shown to be effective for the simulation of biogas upgrading process in up-flow reactors.

Comprehensive evaluation of different strategies to recover methanogenic performance in ammonia-stressed reactors.

In this study, strategies for recovery of ammonia-stressed AD reactors were attempted, by addition of preserved bioaugmentation consortium in gel (BioG), fresh consortium in liquid medium (BioL), woodchip biochar (BW), and straw biochar (BS). In comparison to control group with ammonia, effective treatments, i.e., BioG, BioL, BW and BS raised the maximum methane production rate by 77%, 23%, 35%, and 24%, respectively. BW possibly acted as interspecies electrical conduits for Direct Electron Transfer based on conductivity and SEM analysis. BioG facilitated slow release of bioaugmentation inocula from gel into the AD system, which protected them from a direct environmental shock. According to microbial analysis, both BioG, BioL and BW resulted in increased relative abundance of Methanothermobacter thermautotrophicus; and BS induced selective raise of Methanosarcina thermophila. The increase of methanogens via these strategies led to the faster recovery of the AD process.

Transport and distribution of Salmonella enterica serovar Typhimurium in loamy and sandy soil monoliths with applied liquid manure.

A leaching experiment, where liquid manure spiked with Salmonella enterica serovar Typhimurium (Tet(+)) DSM554 was applied to soil surfaces, was conducted on intact soil monoliths (60 cm in diameter and 100 cm long). A total of 6.5 x 10(10) CFU was applied to each column. We found that Salmonella serovar Typhimurium could be transported to a 1-m depth in loamy soil at concentrations reaching 1.3 x 10(5) CFU/ml of leachate. The test strain was found in concentrations ranging from 300 to 1.3(5) cells/ml in loamy soil throughout the 27 days of the experiment, while concentrations below 20 cells/ml were sporadically detected in the leachates from sandy monoliths. Real-time PCR targeting invA DNA showed a clear correspondence between the total and culturable numbers of cells in the leachate, indicating that most cells leached were viable. On day 28, distribution of Salmonella serovar Typhimurium at five depths in the four monoliths was determined. The highest recovery rate, ranging from 1.5% to 3.8% of the total applied inoculum, was found in the top 0.2 m.

A comparison of the fate of diflufenican in agricultural sandy soil and gravel used in urban areas.

Diflufenican is used in both agricultural and urban areas to control weeds. However, in Europe pesticides are regulated using agricultural soil data only. Urban soils where the top layer is replaced by gravel (e.g. driveways, outdoor tiled areas) can evidently differ from agricultural soils in many biotic and physical properties. In the present study, we compared the degradation, mineralization, sorption and aging of diflufenican between an agricultural sandy soil to a gravel used in urban areas. Both diflufenican and its two main aerobic metabolites were investigated. Diflufenican and the metabolites degraded slower in gravel than in agricultural soil. One of the metabolites, 2-[3-(Trifluoromethyl)phenoxy]nicotinic acid (AE B107137 as identified by EFSA; further abbreviated as AE-B), was formed from the incubation of diflufenican in both soil and gravel, however, showing different formation patterns in the two materials: No accumulation of AE-B was determined in the soil, whereas in gravel, an accumulation of AE-B was determined over the full study period of 150 days. After 150 days, approximately 10% of the applied diflufenican was mineralised in the soil (cumulative), while it was not mineralised in the gravel. Diflufenican showed much stronger sorption to the soil than to the gravel, while the sorption of the metabolites was weaker than diflufenican in both soil and gravel. Within the experimental period, the influence of aging on the fate of diflufenican in soil and gravel is limited (<0.9 and <1.4%, respectively) when compared to the amount of compound still present in the soil. Overall, the results imply shortcomings in the risk assessment procedures requested for the registration of pesticides for urban areas.

TaqMan probe-based real-time PCR assay for detection and discrimination of class I, II, and III tfdA genes in soils treated with phenoxy acid herbicides.

Separate quantification of three classes of tfdA genes was performed using TaqMan quantitative real-time PCR for 13 different soils subsequent to mineralization of three phenoxy acids. Class III tfdA genes were found to be involved in mineralization more often than class I and II tfdA genes.