Utility of wood ash, paper sludge and biochar for the mitigation of greenhouse gases emissions from acid boreal soils.

Land-use change in the boreal forest region leads to agriculture to be carried out on acid, shallow and low fertility soils. To correct soil fertility and promote crop productivity, manure and inorganic fertilizers are added, increasing the risk for greenhouse gas emissions (GHGE). To reduce the reliance on synthetic soil amendments while taking advantage of locally relevant industrial by-products, wood ash (WA) and paper sludge (SL) have been proposed as soil amendments for a range of soil types. We posited that amending soils with WA, SL and biochar will improve soil health parameters and fertility of boreal soils. Microbial activity resulting to nitrogen losses and availability were assessed. WA had a stronger ameliorative effect on acid boreal soils compared to SL. Both WA and SL increased soil microbial biomass and basal respiration resulting in higher net mineralization and thus significant increases in CO2 emissions. Co-application of urea with WA or SL further increased net mineralization compared to when used independently. Conversely, the biochar's effect was short-lived and not significant. Nevertheless, addition of biochar was shown to reduce CO2 emissions in all cases except in WA amended soils. No significant differences were observed in CH4 emissions across all treatments. WA and SL may improve soil fertility and quality but could also contribute significantly to GHG emissions in acid boreal soils. Further research is recommended to assess the mitigating effect of various biochar feedstocks on GHG emissions when co-applied with WA and SL.

Comparison of Multi-Frequency and Multi-Coil Electromagnetic Induction (EMI) for Mapping Properties in Shallow Podsolic Soils.

Electromagnetic induction (EMI) technique is an established method to measure the apparent electrical conductivity (ECa) of soil as a proxy for its physicochemical properties. Multi-frequency (MF) and multi-coil (MC) are the two types of commercially available EMI sensors. Although the working principles are similar, their theoretical and effective depth of investigation and their resolution capacity can vary. Given the recent emphasis on non-invasive mapping of soil properties, the selection of the most appropriate instrument is critical to support robust relationships between ECa and the targeted properties. In this study, we compared the performance of MC and MF sensors by their ability to define relationships between ECa (i.e., MF-ECa and MC-ECa) and shallow soil properties. Field experiments were conducted under wet and dry conditions on a silage-corn field in western Newfoundland, Canada. Relationships between temporally stable properties, such as texture and bulk density, and temporally variable properties, such as soil water content (SWC), cation exchange capacity (CEC) and pore water electrical conductivity (ECw) were investigated. Results revealed significant (p < 0.05) positive correlations of ECa to silt content, SWC and CEC for both sensors under dry conditions, higher correlated for MC-ECa. Under wet conditions, correlation of MF-ECa to temporally variable properties decreased, particularly to SWC, while the correlations to sand and silt increased. We concluded that the MF sensor is more sensitive to changes in SWC which influenced its ability to map temporally variable properties. The performance of the MC sensor was less affected by variable weather conditions, providing overall stronger correlations to both, temporally stable or variable soil properties for the tested Podzol and hence the more suitable sensor toward various precision agricultural practices.

Nontuberculous mycobacteria in the environment of Hranice Abyss, the world's deepest flooded cave (Hranice karst, Czech Republic).

Nontuberculous mycobacteria (NTM) are widely distributed in the environment. On one hand, they are opportunistic pathogens for humans and animals, and on the other hand, they are effective in biodegradation of some persistent pollutants. Following the recently recorded large abundance of NTM in extreme geothermal environments, the aim of the study was to ascertain the occurrence of NTM in the extreme environment of the water zone of the Hranice Abyss (HA). The HA mineral water is acidic, with large concentrations of free CO2, and bacterial slimes creating characteristic mucilaginous formations. Both culture and molecular methods were used to compare the mycobacterial diversity across the linked but distinct ecosystems of HA and the adjacent Zbrasov Aragonite Caves (ZAC) with consideration of their pathogenic relevance. Six slowly growing NTM species (M. arupense, M. avium, M. florentinum, M. gordonae, M. intracellulare) and two rapidly growing NTM species (M. mucogenicum, M. sediminis) were identified in the water and in the dry zones at both sites. Proteobacteria were dominant in all the samples from both the HA and the ZAC. The bacterial microbiomes of the HA mineral water and HA slime were similar, but both differed from the microbiome in the ZAC mineral water. Actinobacteria, a phylum containing mycobacteria, was identified in all the samples at low proportional abundance. The majority of the detected NTM species belong among environmental opportunistic pathogens.

Northward shift of the agricultural climate zone under 21st-century global climate change.

As agricultural regions are threatened by climate change, warming of high latitude regions and increasing food demands may lead to northward expansion of global agriculture. While socio-economic demands and edaphic conditions may govern the expansion, climate is a key limiting factor. Extant literature on future crop projections considers established agricultural regions and is mainly temperature based. We employed growing degree days (GDD), as the physiological link between temperature and crop growth, to assess the global northward shift of agricultural climate zones under 21st-century climate change. Using ClimGen scenarios for seven global climate models (GCMs), based on greenhouse gas (GHG) emissions and transient GHGs, we delineated the future extent of GDD areas, feasible for small cereals, and assessed the projected changes in rainfall and potential evapotranspiration. By 2099, roughly 76% (55% to 89%) of the boreal region might reach crop feasible GDD conditions, compared to the current 32%. The leading edge of the feasible GDD will shift northwards up to 1200 km by 2099 while the altitudinal shift remains marginal. However, most of the newly gained areas are associated with highly seasonal and monthly variations in climatic water balances, a critical component of any future land-use and management decisions.

Using agro-industrial wastes for the cultivation of microalgae and duckweeds: Contamination risks and biomass safety concerns.

Aquatic organisms, such as microalgae (Chlorella, Arthrospira (Spirulina), Tetrasselmis, Dunalliela etc.) and duckweed (Lemna spp., Wolffia spp. etc.) are a potential source for the production of protein-rich biomass and for numerous other high-value compounds (fatty acids, pigments, vitamins etc.). Their cultivation using agro-industrial wastes and wastewater (WaW) is of particular interest in the context of a circular economy, not only for recycling valuable nutrients but also for reducing the requirements for fresh water for the production of biomass. Recovery and recycling of nutrients is an unavoidable long-term approach for securing future food and feed production. Agro-industrial WaW are rich in nutrients and have been widely considered as a potential nutrient source for the cultivation of microalgae/duckweed. However, they commonly contain various hazardous contaminants, which could potentially taint the produced biomass, raising various concerns about the safety of their consumption. Herein, an overview of the most important contaminants, including heavy metals and metalloids, pathogens (bacteria, viruses, parasites etc.), and xenobiotics (hormones, antibiotics, parasiticides etc.) is given. It is concluded that pretreatment and processing of WaW is a requisite step for the removal of several contaminants. Among the various technologies, anaerobic digestion (AD) is widely used in practice and offers a technologically mature approach for WaW treatment. During AD, various organic and biological contaminants are significantly removed. Further removal of contaminants could be achieved by post-treatment and processing of digestates (solid/liquid separation, dilution etc.) to further decrease the concentration of contaminants. Moreover, during cultivation an additional removal may occur through various mechanisms, such as precipitation, degradation, and biotransformation. Since many jurisdictions regulate the presence of various contaminants in feed or food setting strict safety monitoring processes, it would be of particular interest to initiate a multi-disciplinary discussion whether agro-industrial WaW ought to be used to cultivate microalgae/duckweed for feed or food production and identify most feasible options for doing this safely. Based on the current body of knowledge it is estimated that AD and post-treatment of WaW can lower significantly the risks associated with heavy metals and pathogens, but it is yet unclear to what extent this is the case for certain persistent xenobiotics.

The influence of flint stones on a soil microbial community in the northern Negev Desert.

In the Negev Desert ecosystems, flint-stone cover on slopes acts as a barrier against water flow. As a result, soil moisture increases and organic matter accumulates under the stone and in the immediate surroundings, both affecting the duration of soil microbial activity. On the other hand, during the dry season (characterized by approximately 210 dew nights), flint-stone cover plays an important role in the formation of dew, which eventually trickles down beneath the stone, correspondingly enhancing biological activity. The present study examined the possible role of flint stones as hotspots for soil microbial-community activity and diversity. The results were compared with those obtained from the adjacent stone-free soils in the open spaces (OS), which served as controls. Microbial activity (respiration and biomass) and functional diversity were determined by the MicroResp™ method. In addition, estimates of genetic diversity and viable counts of bacteria and fungi [colony-forming units (CFUs)] were obtained. The soil was significantly wetter and contained more organic matter beneath the flint stones (BFS). As hypothesized, biological activity was enhanced under the stones, as described by CO2 evolution, microbial-community biomass functional diversity, and fungal phylogenetic diversity. BFS environments favored a greater range of catabolic functions. Taxa generally known for their stress resilience were found in the OS habitats. The results of this study elucidate the importance of flint-stone cover to soil microbial biomass, community activity, and functional diversity in the northern Negev Desert.

Desert Perennial Shrubs Shape the Microbial-Community Miscellany in Laimosphere and Phyllosphere Space.

Microbial function, composition, and distribution play a fundamental role in ecosystem ecology. The interaction between desert plants and their associated microbes is expected to greatly affect their response to changes in this harsh environment. Using comparative analyses, we studied the impact of three desert shrubs, Atriplex halimus (A), Artemisia herba-alba (AHA), and Hammada scoparia (HS), on soil- and leaf-associated microbial communities. DNA extracted from the leaf surface and soil samples collected beneath the shrubs were used to study associated microbial diversity using a sequencing survey of variable regions of bacterial 16S rRNA and fungal ribosomal internal transcribed spacer (ITS1). We found that the composition of bacterial and fungal orders is plant-type-specific, indicating that each plant type provides a suitable and unique microenvironment. The different adaptive ecophysiological properties of the three plant species and the differential effect on their associated microbial composition point to the role of adaptation in the shaping of microbial diversity. Overall, our findings suggest a link between plant ecophysiological adaptation as a "temporary host" and the biotic-community parameters in extreme xeric environments.

Enzymatic Pre-treatment of Wastewater to Minimize Recovery by Reverse Transcriptase PCR of RNA from Inactive Bacteriophages.

Quantitative viral risk assessments for wastewaters are notoriously difficult. The often considered quantitative reverse transcriptase PCR reflects poorly on virus infectivity rates leading to inaccurate risk interpretations. Various techniques focused on the degradation of the nucleic acids of non-infective viruses were previously employed. We comparatively assessed the effectiveness of such enzymatic treatments for MS2 bacteriophage in treated wastewaters. The single use of RNase A at an appropriate concentration may be as effective as the combination of RNase followed by Proteinase K and more rapid. While all tested enzymatic treatments minimized recovery of RNA (>95 %) in the absence of infective MS2, none completely eliminated the signal recovery. Selection of any enzymatic protocol for minimizing recovery of RNA from degraded, non-infective viruses should balance the methods efficacy with its expediency.

Short RNA indicator sequences are not completely degraded by autoclaving.

Short indicator RNA sequences (<100 bp) persist after autoclaving and are recovered intact by molecular amplification. Primers targeting longer sequences are most likely to produce false positives due to amplification errors easily verified by melting curves analyses. If short indicator RNA sequences are used for virus identification and quantification then post autoclave RNA degradation methodology should be employed, which may include further autoclaving.

Pyrosequencing estimates of the diversity of antibiotic resistant bacteria in a wastewater system.

Standard protocols for monitoring wastewater treatment efficacy target Escherichia coli and fecal coliforms. This might not accurately describe risks associated with antibiotic resistance in the bacterial population of treated wastewaters. We modified a standard agar recovery method by amending it with various antibiotics. The resulting bacterial colonies were submitted to 454 pyrosequencing; thus we identified the diversity of culturable antibiotic resistant bacteria from treated and raw wastewaters. This approach produced 209,706 high quality reads of >300 bp. Operational taxonomic units (OTUs) within Proteobacteria dominated the system. The Shannon-Wiener H' index showed bacterial populations recovered on ciprofloxacin amended agars to be the least diverse. Principal component analysis of OTU distribution at phylum level showed that Proteobacteria accounted for most of the variability. The same analysis revealed most of the samples to have similar diversities at phylum level being dominated by Proteobacteria, though a few samples, typically recovered from ciprofloxacin or doxycycline amended agars were often dissimilar. Arcobacter spp. or E. coli were dominant in the bacterial communities recovered on agars amended with ciprofloxacin or doxycycline, respectively. Genera containing putative pathogens were mostly representatives of Gamma and Epsilon proteobacteria. Bacterial populations containing multiple antibiotic resistance (MAR) in the final treated effluent was a possibility.

Soil microbial diversity in the vicinity of desert shrubs.

Water and nutrient availability are the major limiting factors of biological activity in arid and semiarid ecosystems. Therefore, perennial plants have developed different ecophysiological adaptations to cope with harsh conditions. The chemical profile of the root exudates varies among plant species and this can induce variability in associated microbial populations. We examined the influence of two shrubs species, Artemisia sieberi and Noaea mucronata, on soil microbial diversity. Soil samples were collected monthly, from December 2006 to November 2007, near canopies of both shrubs (0-10-cm depth). Samples were used for abiotic tests and determination of soil bacterial diversity. No significant differences were found in the abiotic variables (soil moisture, total organic matter, and total soluble nitrogen (TSN)) between soil samples collected from under the two shrubs during the study period. No obvious differences in the Shannon-Weaver index, evenness values, or total phylogenetic distances were found for the soil microbial communities. However, detailed denaturing gradient gel electrophoresis (DGGE) clustering as well as taxonomic diversity analyses indicated clear shifts in the soil microbial community composition. These shifts were governed by seasonal variability in water availability and, significantly, by plant species type.

Microarray assessment of virulence, antibiotic, and heavy metal resistance in an agricultural watershed creek.

Potential risks associated with impaired surface water quality have commonly been evaluated by indirect description of potential sources using various fecal microbial indicators and derived source-tracking methods. These approaches are valuable for assessing and monitoring the impacts of land-use changes and changes in management practices at the source of contamination. A more detailed evaluation of putative etiologically significant genetic determinants can add value to these assessments. We evaluated the utility of using a microarray that integrates virulence genes with antibiotic and heavy metal resistance genes to describe and discriminate among spatially and seasonally distinct water samples from an agricultural watershed creek in Eastern Ontario. Because microarray signals may be analyzed as binomial distributions, the significance of ambiguous signals can be easily evaluated by using available off-the-shelf software. The FAMD software was used to evaluate uncertainties in the signal data. Analysis of multilocus fingerprinting data sets containing missing data has shown that, for the tested system, any variability in microarray signals had a marginal effect on data interpretation. For the tested watershed, results suggest that in general the wet fall season increased the downstream detection of virulence and resistance genes. Thus, the tested microarray technique has the potential to rapidly describe the quality of surface waters and thus to provide a qualitative tool to augment quantitative microbial risk assessments.

Endophyte microbiome diversity in micropropagated Atriplex canescens and Atriplex torreyi var griffithsii.

Microbial diversity associated with micropropagated Atriplex species was assessed using microscopy, isolate culturing, and sequencing. Light, electron, and confocal microscopy revealed microbial cells in aseptically regenerated leaves and roots. Clone libraries and tag-encoded FLX amplicon pyrosequencing (TEFAP) analysis amplified sequences from callus homologous to diverse fungal and bacterial taxa. Culturing isolated some seed borne endophyte taxa which could be readily propagated apart from the host. Microbial cells were observed within biofilm-like residues associated with plant cell surfaces and intercellular spaces. Various universal primers amplified both plant and microbial sequences, with different primers revealing different patterns of fungal diversity. Bacterial and fungal TEFAP followed by alignment with sequences from curated databases revealed 7 bacterial and 17 ascomycete taxa in A. canescens, and 5 bacterial taxa in A. torreyi. Additional diversity was observed among isolates and clone libraries. Micropropagated Atriplex retains a complex, intimately associated microbiome which includes diverse strains well poised to interact in manners that influence host physiology. Microbiome analysis was facilitated by high throughput sequencing methods, but primer biases continue to limit recovery of diverse sequences from even moderately complex communities.

Recovery of Escherichia coli from soil after addition of sterile organic wastes.

Laboratory batch tests indicate that addition of sterile municipal sewage biosolids to clay soil from four depths increases the numbers of Escherichia coli isolates recoverable in EC-MUG broth (EC broth with 4-methylumbelliferyl-beta-glucuronide). This effect was most marked for the deeper soil layers, with increases of about 2.6 orders of magnitude in E. coli most probable number.