Nanoparticles within WWTP sludges have minimal impact on leachate quality and soil microbial community structure and function

One of the main pathways by which engineered nanoparticles (ENPs) enter the environment is through land application of waste water treatment plant (WWTP) sewage sludges. WWTP sludges, enriched with Ag and ZnO ENPs or their corresponding soluble metal salts during anaerobic digestion and subsequently mixed with soil (targeting a final concentration of 1400 and 140 mg/kg for Zn and Ag, respectively), were subjected to 6 months of ageing and leaching in lysimeter columns outdoors. Amounts of Zn and Ag leached were very low, accounting for <0.3% and <1.4% of the total Zn and Ag, respectively. No differences in total leaching rates were observed between treatments of Zn or Ag originally input to WWTP as ENP or salt forms. Phospholipid fatty acid profiling indicated a reduction in the fungal component of the soil microbial community upon metal exposure. However, overall, the leachate composition and response of the soil microbial community following addition of sewage sludge enriched either with ENPs or metal salts was very similar

Denitrification likely catalyzed by endobionts in an allogromiid foraminifer

Author Posting. © The Author(s), 2011. This is the author's version of the work. It is posted here by permission of Nature Publishing Group for personal use, not for redistribution. The definitive version was published in The ISME Journal 6 (2012): 951–960, doi:10.1038/ismej.2011.171.Nitrogen can be a limiting macronutrient for carbon uptake by the marine biosphere. The process of denitrification (conversion of nitrate to gaseous compounds, including N2) removes bioavailable nitrogen, particularly in marine sediments, making it a key factor in the marine nitrogen budget. Benthic foraminifera reportedly perform complete denitrification, a process previously considered nearly exclusively performed by bacteria and archaea. If the ability to denitrify is widespread among these diverse and abundant protists, a paradigm shift is required for biogeochemistry and marine microbial ecology. However, to date, the mechanisms of foraminiferal denitrification are unclear and it is possible that the ability to perform complete denitrification is due to symbiont metabolism in some foraminiferal species. Using sequence analysis and GeneFISH, we show that for a symbiont-bearing foraminifer, the potential for denitrification resides in the endobionts. Results also identify the endobionts as denitrifying pseudomonads and show that the allogromiid accumulates nitrate intracellularly, presumably for use in denitrification. Endobionts have been observed within many foraminiferal species, and in the case of associations with denitrifying bacteria, may provide fitness for survival in anoxic conditions. These associations may have been a driving force for early foraminiferal diversification, which is thought to have occurred in the Neoproterozoic when anoxia was widespread.This research was supported by NSF grant EF-0702491 to JMB, KLC and VPE; some ship support was provided by NSF MCB-0604084 to VPE and JMB.2012-06-0

Compound driven differences in N2 and N2O emission from soil; the role of substrate use efficiency and the microbial community

Organic C is an important control on the process of denitrification, a process that can result in the production and reduction of the potent greenhouse gas nitrous oxide (N2O). This study identified the influence of different low molecular weight C (LMW-C) compounds on the production of nitrous oxide (N2O) and dinitrogen (N2) and the associated role of the size and structure of the microbial community. We examined this following application of glucose, glutamine or citric acid (250 mg C kg−1 dry soil) and 15N-KNO3 (100 mg N kg−1 dry soil) to a sandy loam soil and measured the production of N2 and N2O by denitrifiers using 15N labeling techniques, changes in the bacterial community as measured by T-RFLP on 16SrDNA fragments and changes in the gene copy number of 16SrDNA, nirK, nirS and nosZ over 144 h. Addition of glucose, citric acid and glutamine all increased emissions of 15N-N2 above that found in the control (P < 0.05) while the addition of glucose and glutamine resulted in higher emissions of 14+15N-N2O (P < 0.001) than the addition of citric acid, resulting in a lower 15N-N2O to 15N-N2 ratio in the citric acid treatment. The 16SrDNA gene copy number increased after addition of citric acid and glutamine, whilst 16SrDNA showed significant shifts in community composition in all C treatments although over different time periods. The gene copy number of nosZ only significantly increased at 120 h in the glutamine treatment (P < 0.05) and nirS at 120 h in the citric acid and glutamine treatments (P < 0.05). This suggests that where C is added as a single input, differences in N2 and N2O emissions between LMW-C compounds were not caused by selection for denitrifiers but likely driven by differences in substrate use efficiency and subsequent differences in C partitioning between growth and respiration. The differing influence of the three selected C compounds on denitrification indicates the potential for lowering net N2O emissions through regulation of C compound availability

The incidence of nirS and nirK and their genetic heterogeneity in cultivated denitrifiers

Gene sequence analysis of nirS and nirK, both encoding nitrite reductases, was performed on cultivated denitrifiers to assess their incidence in different bacterial taxa and their taxonomical value. Almost half of the 227 investigated denitrifying strains did not render an nir amplicon with any of five previously described primers. NirK and nirS were found to be prevalent in Alphaproteobacteria and Betaproteobacteria, respectively, nirK was detected in the Firmicutes and Bacteroidetes and nirS and nirK with equal frequency in the Gammaproteobacteria. These observations deviated from the hitherto reported incidence of nir genes in bacterial taxa. NirS gene phylogeny was congruent with the 16S rRNA gene phylogeny on family or genus level, although some strains did group within clusters of other bacterial classes. Phylogenetic nirK gene sequence analysis was incongruent with the 16S rRNA gene phylogeny. NirK sequences were also found to be significantly more similar to nirK sequences from the same habitat than to nirK sequences retrieved from highly related taxa. This study supports the hypothesis that horizontal gene transfer events of denitrification genes have occurred and underlines that denitrification genes should not be linked with organism diversity of denitrifiers in cultivation-independent studies

The effect of soil properties on the toxicity of silver to the soil nitrification process

Silver (Ag) is being increasingly used in a range of consumer products, predominantly as an antimicrobial agent, leading to a higher likelihood of its release into the environment. The present study investigated the toxicity of Ag to the nitrification process in European and Australian soils in both leached and unleached conditions. Overall, leaching of soils was found to have a minimal effect on the final toxicity data, with an average leaching factor of approximately 1. Across the soils, the toxicity was found to vary by several orders of magnitude, with concentrations of Ag causing a 50% reduction in nitrification relative to the controls (EC50) ranging from 0.43 mg Ag/kg to >640 mg Ag/kg. Interestingly, the dose-response relationships in most of the soils showed significant stimulation in nitrification at low Ag concentrations (i.e., hormesis), which in some cases produced responses up to double that observed in the controls. Soil pH and organic carbon were the properties found to have the greatest influence on the variations in toxicity thresholds across the soils, and significant relationships were developed that accounted for approximately 90% of the variability in the data. The toxicity relationships developed from the present study will assist in future assessment of potential Ag risks and enable the site-specific prediction of Ag toxicity.Kate A. Langdon, Mike J. Mclaughlin, Jason K. Kirby, and Graham Merringto