Subsurface Manure Injection Reduces Surface Transport of Antibiotic Resistance Genes but May Create Antibiotic Resistance Hotspots in Soils.

Compared to surface application, manure subsurface injection reduces surface runoff of nutrients, antibiotic resistant microorganisms, and emerging contaminants. Less is known regarding the impact of both manure application methods on surface transport of antibiotic resistance genes (ARGs) in manure-amended fields. We applied liquid dairy manure to field plots by surface application and subsurface injection and simulated rainfall on the first or seventh day following application. The ARG richness, relative abundance (normalized to 16s rRNA), and ARG profiles in soil and surface runoff were monitored using shotgun metagenomic sequencing. Within 1 day of manure application, compared to unamended soils, soils treated with manure had 32.5-70.5% greater ARG richness and higher relative abundances of sulfonamide (6.5-129%) and tetracycline (752-3766%) resistance genes (p ≤ 0.05). On day 7, soil ARG profiles in the surface-applied plots were similar to, whereas subsurface injection profiles were different from, that of the unamended soils. Forty-six days after manure application, the soil ARG profiles in manure injection slits were 37% more diverse than that of the unamended plots. The abundance of manure-associated ARGs were lower in surface runoff from manure subsurface injected plots and carried a lower resistome risk score in comparison to surface-applied plots. This study demonstrated, for the first time, that although manure subsurface injection reduces ARGs in the runoff, it can create potential long-term hotspots for elevated ARGs within injection slits.

A spectrum of preferential flow alters solute mobility in soils.

Preferential flow reduces water residence times and allows rapid transport of pollutants such as organic contaminants. Thus, preferential flow is considered to reduce the influence of soil matrix-solute interactions during solute transport. While this claim may be true when rainfall directly follows solute application, forcing rapid chemical and physical disequilibrium, it has been perpetuated as a general feature of solute transport-regardless of the magnitude preferential flow. A small number of studies have alternatively shown that preferential transport of strongly sorbing solutes is reduced when solutes have time to diffuse and equilibrate within the soil matrix. Here we expand this inference by allowing solute sorption equilibrium to occur and exploring how physiochemical properties affect solute transport across a vast range of preferential flow. We applied deuterium-labeled rainfall to field plots containing manure spiked with eight common antibiotics with a range of affinity for the soil after 7 days of equilibration with the soil matrix and quantified preferential flow and solute transport using 48 soil pore water samplers spread along a hillslope. Based on > 700 measurements, our data showed that solute transport to lysimeters was similar-regardless of antibiotic affinity for soil-when preferential flow represented less than 15% of the total water flow. When preferential flow exceeded 15%, however, concentrations were higher for compounds with relatively low affinity for soil. We provide evidence that (1) bypassing water flow can select for compounds that are more easily released from the soil matrix, and (2) this phenomenon becomes more evident as the magnitude of preferential flow increases. We argue that considering the natural spectrum preferential flow as an explanatory variable to gauge the influence of soil matrix-solute interactions may improve parsimonious transport models.