Application of denitrifying bioreactors for the removal of atrazine in agricultural drainage water.

Atrazine and nitrate NO3-N are two agricultural pollutants that occur widely in surface and groundwater. One of the pathways by which these pollutants reach surface water is through subsurface drainage tile lines. Edge-of-field anaerobic denitrifying bioreactors apply organic substrates such as woodchips to stimulate the removal of NO3-N from the subsurface tile waters through denitrification. Here we investigated the co-removal of NO3-N and atrazine by these bioreactors. Laboratory experiments were conducted using 12-L woodchips-containing flow-through bioreactors, with and without the addition of biochar, to treat two concentrations of atrazine (20 and 50 µg L-1) and NO3-N (1.5 and 11.5 mg L-1), operated at four hydraulic retention time, HRT, (4 h, 8 h, 24 h, 72 h). Additionally, we examined the effect of aerating the bioreactors on atrazine removal. Furthermore, we tested atrazine removal by a field woodchip denitrifying bioreactor. The removal of both NO3-N and atrazine increased with increasing HRT in the laboratory bioreactors. At 4 h, the woodchip bioreactors removed 65% of NO3-N and 25% of atrazine but, at 72 h, the bioreactors eliminated all the NO3-N and 53% of atrazine. Biochar-amended bioreactors removed up to 90% of atrazine at 72-h retention time. We concluded that atrazine removal was primarily via adsorption because neither aeration nor NO3-N levels had an effect. At 4-h retention time, the field bioreactors achieved 2.5 times greater atrazine removal than the laboratory bioreactors. Our findings thus highlighted hydraulic retention time and biochar amendments as two important factors that may control the efficiency of atrazine removal by denitrifying bioreactors. In sum, laboratory and field data demonstrated that denitrifying bioreactors have the potential to decrease pesticide transport from agricultural lands to surface waters.

Seasonal performance of denitrifying bioreactors in the Northeastern United States: Field trials.

Denitrifying bioreactors are increasingly being used for nitrate removal from agricultural drainage water. Filled with carbon substrates, often woodchips, denitrifying bioreactors provide a favorable anaerobic environment for denitrification. Despite performing well in loess soils in the Midwestern United States, field bioreactors have not yet been evaluated in shallow soils over glacial till that are characteristic for the Northeastern United States. This study, therefore, investigates the performance of bioreactors and provides design criteria for shallow soil with flashy discharges. Paired bioreactors, one filled with woodchips and one with a mixture of woodchip and biochar, were installed in tile drained fields in three landscapes in New York State. The bioreactors were monitored for a three-year period during which, the flow rate, temperature, nitrate (NO3--N), sulfate (SO42--S) and dissolved organic carbon (DOC) were measured. Results showed that the average NO3--N removal efficiency during the three years of observations was about 50%. The NO3--N removal rate ranged from 0 in winter to 72 g d-1 m-3 in summer. We found that biochar was only effective during the first year in enhancing denitrification, due to the ageing. An index for carbon availability related to NO3--N removal was developed. During winter, availability of the DOC was a limiting factor in bioreactor performance. Finally, to aid in the design of bioreactors, we developed generalizable relationships between the removal efficiency and hydraulic retention time and temperature.

Physiological and evolutionary contexts of a new symbiotic species from the nitrogen-recycling gut community of turtle ants.

While genome sequencing has expanded our knowledge of symbiosis, role assignment within multi-species microbiomes remains challenging due to genomic redundancy and the uncertainties of in vivo impacts. We address such questions, here, for a specialized nitrogen (N) recycling microbiome of turtle ants, describing a new genus and species of gut symbiont-Ischyrobacter davidsoniae (Betaproteobacteria: Burkholderiales: Alcaligenaceae)-and its in vivo physiological context. A re-analysis of amplicon sequencing data, with precisely assigned Ischyrobacter reads, revealed a seemingly ubiquitous distribution across the turtle ant genus Cephalotes, suggesting ≥50 million years since domestication. Through new genome sequencing, we also show that divergent I. davidsoniae lineages are conserved in their uricolytic and urea-generating capacities. With phylogenetically refined definitions of Ischyrobacter and separately domesticated Burkholderiales symbionts, our FISH microscopy revealed a distinct niche for I. davidsoniae, with dense populations at the anterior ileum. Being positioned at the site of host N-waste delivery, in vivo metatranscriptomics and metabolomics further implicate I. davidsoniae within a symbiont-autonomous N-recycling pathway. While encoding much of this pathway, I. davidsoniae expressed only a subset of the requisite steps in mature adult workers, including the penultimate step deriving urea from allantoate. The remaining steps were expressed by other specialized gut symbionts. Collectively, this assemblage converts inosine, made from midgut symbionts, into urea and ammonia in the hindgut. With urea supporting host amino acid budgets and cuticle synthesis, and with the ancient nature of other active N-recyclers discovered here, I. davidsoniae emerges as a central player in a conserved and impactful, multipartite symbiosis.

Metabolomics analysis of unresolved molecular variability in stoichiometry dynamics of a stream dissolved organic matter.

Broad molecular classification based on stoichiometric ratio relationships has been used extensively to characterize the chemical diversity of aquatic dissolved organic matter (DOM). However, variability in the molecular composition within this classification has remained elusive, thus limiting the interpretation of DOM dynamics, especially with respect to transport versus transformation patterns in response to hydrologic or landscape changes. Here, leveraging high-frequency spatiotemporal sampling during rainfall events at a Critical Zone Observatory project site in Clear Creek, Iowa, we apply a metabolomics-based analysis validated with fragmentation using tandem mass spectrometry to uncover patterns in the molecular features of the DOM composition that were not resolved by classification based on stoichiometric ratios in the chemical formulae. From upstream to downstream sites, beyond the increased aromaticity implied by changes in the stoichiometric ratios, we identified an increased abundance of flavonoids and other phenylpropanoids, two important subgroups of aromatic compounds. The stoichiometric analysis also proposed a localized decline in the abundance of lipid-like compounds, which we attributed specifically to medium-chain and short-chain fatty acids; other lipids such as long-chain fatty acids and sterol lipids remained unchanged. We further determined in-stream molecular transitions and specific compound degradation by capturing changes in the molecular masses of terpenoids, phenylpropanoids, fatty acids, and amino acids. In sum, the metabolomics analysis of the chemical formulae resolved molecular variability imprinted on the stoichiometric DOM composition to implicate key molecular subgroups underlying carbon transport and cycling dynamics in the stream.