Heavy metal could drive co-selection of antibiotic resistance in terrestrial subsurface soils.

Terrestrial surface ecosystems are important sinks for antibiotic resistance genes (ARGs) due to the continuous discharge of contaminants from human-impacted ecosystems. However, the abundance and resistance types of ARGs and their influencing factors in terrestrial subsurface soils are not well known. In this study, we investigated the abundance and diversity of ARGs, and their correlations with metal resistance genes (MRGs), mobile genetic elements (MGEs), bacteria, and heavy metals in subsurface soils using high throughput quantitative PCR and metagenomic sequencing approaches. Abundant and diverse ARGs were detected with high spatial heterogeneity among sampling sites. Vertically, there was no significant difference in ARG profiles between the aquifer and non-aquifer soils. Heavy metals were key factors shaping ARG profiles in soils with high heavy metal contents, while they showed no significant effect in low contents. Moreover, heavy metals could trigger the proliferation of antibiotic resistance by increasing MGE abundance or influencing bacterial communities. Metagenomic analysis also revealed the widespread co-occurrence of ARGs and MRGs, with heavy metals possibly enhancing the co-selection of ARGs and MRGs in soils with high heavy metal contents. This study highlighted the heavy metal-driven co-selection of ARGs and revealed the occurrence of ARG pollution in terrestrial subsurface soils.

Source, contribution and microbial N-cycle of N-compounds in China fresh snow.

The importance and contribution of nitrogen compounds and the related microbial nitrogen cycling processes in fresh snow are not well understood under the current research background. We collected fresh snow samples from 21 cities that 80% are from China during 2016 and 2017. Principal component analysis showed that SO42- were in the first principal component, and N-compounds were the second. Furthermore, the main pollutant ions SO42- and NO3- were from anthropogenic sources, and SO42- contributed (61%) more to the pollution load than NO3- (29%), which were confirmed through a series of precipitation mechanism analysis. We selected five N-cycle processes (consist of oxidation and reduction processes) for molecular biology experiments, including Ammonia-oxidation process, Nitrite-oxidation process, Denitrification process, Anaerobic-ammoxidation process (Anammox) and Dissimilatory nitrate reduction to ammonium process (DNRA). Except ammonia-oxidizing archaeal (AOA) and bacterial (AOB) amoA genes (above 107 copies g-1), molecular assays of key functional genes in various nitrogen conversion processes showed a belowed detection limit number, and AOB abundance was always higher than AOA. The determination of the microbial transformation rate using the 15N-isotope tracer technique showed that the potential rate of five N-conversion processes was very low, which is basically consistent with the results from molecular biology studies. Taken together, our results illustrated that microbial nitrogen cycle processes are not the primary biological processes causing the pollution in China fresh snow.

Effects of hydraulic loading rate and aeration mode on nitrogen removal and nitrogen functional gene abundances in subsurface wastewater infiltration systems.

Matrix dissolved oxygen, nitrogen removal and nitrogen functional gene abundances in two artificial aeration modes, continuous aeration (CA) and intermittent aeration (IA), in subsurface wastewater infiltration systems (SWISs) under different hydraulic loading rates (HLRs) were investigated. Aeration not only successfully created aerobic conditions at 50 cm depth, but also did not change anoxic or anaerobic conditions at 80 and 110 cm depths. Meanwhile, aeration significantly enhanced chemical oxygen demand, NH4+-N, and total nitrogen (TN) removal and the enrichment of nitrogen removal functional genes (amoA, nxrA, napA, narG, nirK and qnorB) compared to the non-aerated SWIS, especially for high HLRs. IA SWIS (79.7%-85.8%) had a better performance on TN removal compared with CA SWIS (73.8%-82.2%) when the HLRs ranged from 0.06 to 0.3 m3/(m2 d). Intermittent aeration is a sensible strategy to achieve high HLR, good nitrogen removal performance and comparatively low operation cost for SWISs.

Effect of COD/N Ratio on Removal Performances in Two Subsurface Wastewater Infiltration Systems.

Dissolved oxygen (DO), were investigated. Aerobic conditions were effectively developed in 50 cm depth of the matrix and anoxic or anaerobic conditions were not changed in 80 and 110 cm depth by intermittent aeration, which encouraged nitrification. Increased influent COD/N ratio led to lower COD and nitrogen removal in conventional SWISs. Sufficient carbon source in high COD/N ratio influent promoted denitrification with intermittent aeration. High removal rates of COD (95.68 ± 0.21%), TP (92.02 ± 0.28%), -N (99.33 ± 0.05%), and - (89.65 ± 0.6%) were obtained with influent COD/N ratio of 12 in aerated SWISs. Under the COD/N ratio of 12 and 18, intermittent aeration boosted the growth and reproduction of nitrifying bacteria and denitrifying bacteria. Meanwhile, nitrate and nitrite reductase activities with intermittent aeration were higher than that without aeration in 80 and 110 cm depths.

Performance of organics and nitrogen removal in subsurface wastewater infiltration systems by intermittent aeration and shunt distributing wastewater.

Organics and nitrogen removal in four subsurface wastewater infiltration systems (SWISs), named SWIS A (without intermittent aeration and shunt distributing wastewater), SWIS B (with intermittent aeration), SWIS C (with shunt distributing wastewater) and SWIS D (with intermittent aeration and shunt distributing wastewater) was investigated. High average removal rates of 92.3% for COD, 90.2% for NH4-N and 88.1% for TN were achieved simultaneously in SWIS D compared with SWIS A, B and C. The excellent TN removal of SWIS D was due to intermittent aeration provided sufficient oxygen for nitrification in upper matrix and the favorable anoxic or anaerobic environment for denitrification in subsequent matrix, and moreover, shunt distributing wastewater provided sufficient carbon source for denitrification process. The results indicated that intermittent artificial aeration combined with shunt distributing wastewater could achieve high organics and nitrogen removal in SWISs.

Effects of intermittent aeration on pollutants removal in subsurface wastewater infiltration system.

In this study, the pollutant removal performances in two pilot-scale subsurface wastewater infiltration systems (SWISs) with and without intermittent aeration were investigated. Matrix oxidation reduction potential (ORP) results showed that intermittent aeration well developed aerobic conditions in upper matrix and anoxic or anaerobic conditions in the subsequent sections, which resulted in high NH4(+)-N and TN removal. Moreover, intermittent aeration increased removal rates of COD and TP. Microbial populations and enzyme activities analysis proved that intermittent aeration not only obviously boosted the growth and reproduction of bacteria, fungus, actinomyces, nitrifying bacteria and denitrifying bacteria, but also successfully increased nitrate reductase (NR) and nitrite reductase (NIR) in the depth of 80 and 110 cm. The results suggest that the intermittent aeration could be a widespread research and application strategy for achieving the high removal performance in SWISs.