Effects of antibiotics on soil microorganisms: time and nutrients influence pollution-induced community tolerance

A method for the detection of the effects of antibiotics on soil microbial communities was optimised in the present study. We investigated the influence of measurement time and nutrient status on the pollution-induced community tolerance (PICT), using the sulfonamide sulfachloropyridazine (SCP) as model compound. The tolerance development in soil microcosms that were exposed to SCP under different conditions was compared with the background tolerance in SCP-unexposed microcosms. The tolerance of bacterial extracts from the soil microcosms was determined in Biolog® multiwell plates as the SCP sensitivity of a range of physiological processes. The background tolerance was not affected by soil nutrient amendments, but an influence of the inoculum density in the microtiter plates was observed. Still, the variance of the background tolerance was low, which is in favour of the usage of community tolerance measurements for a selective detection of a toxicant impact. In line with the assumptions of the PICT concept, soil amendment with SCP led to an increase in community tolerance. This tolerance development was enhanced upon additional soil amendment with fresh pig slurry, and less by alfalfa meal addition. Tolerance increases were observed after a soil exposure to SCP of only 7 days, possibly because nutrient input facilitates the fast adaptation of the soil microbial community. However, a further increase in exposure time led to variable changes in the observed tolerance. Prolonged tolerance detection in the microtiter plates (11 days) enabled a clearer differentiation between different soil treatments, as it better resolved the EC50 values of processes with a high tolerance to bacteriostatic antimicrobial compounds. For the detection of antibiotic effects on soil microbial communities, it is therefore recommended to use nutrient amendments (possibly fresh pig slurry), to standardize the soil exposure time, and to extend the period of Biolog plate measurement beyond 7 days.