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A laboratory work has been carried out to determine the tolerance of soil bacterial communities to Ni and Zn and co-tolerance to tetracycline antibiotics (chlortetracycline (CTC), oxytetracycline (OTC) and tetracycline (TC)) in soils individually spiked with five different concentrations of Ni or Zn (1,000, 750, 500, 250, and 125 mg kg−1), and an uncontaminated (0 mg kg−1) control soil. The PICT parameter (pollution-induced community tolerance) was estimated for the bacterial community using the tritium (3H)-labeled leucine incorporation technique, and the values corresponding to log IC50 were used as toxicity index. The mean log IC50 values observed in the uncontaminated soil samples indicate that Zn (with log IC50 = −2.83) was more toxic than Ni (log IC50 = −2.73). In addition, for the soil with the lowest carbon content (C = 1.9%), Ni-contaminated samples showed increased tolerance when the Ni concentrations added were ≥500 mg kg−1, while for the soils with higher carbon content (between 5.3% and 10.9%) tolerance increased when Ni concentrations added were ≥1,000 mg kg−1. Regarding the soils contaminated with Zn, tolerance increased in all the soils studied when the Zn concentrations added were ≥125 mg kg−1, regardless of the soil carbon content. The co-tolerance increases obtained after exposure of the bacterial suspension to TC, OTC and CTC showed an identical behavior within these tetracycline antibiotics. However, it was dependent on the heavy metal tested (Ni or Zn). In the case of soils 1 (C = 1.1%) and 2 (C = 5.3%), the soil bacterial communities showed increases in co-tolerance to TC, OTC and CTC for Ni concentrations added of ≥125 mg kg−1, while for soil 3 (with C = 10.9%) co-tolerance took place when Ni was added at ≥1,000 mg kg−1. However, in soils contaminated with Zn, increases in co-tolerance to CTC, OTC and TC occurred at Zn concentrations added of ≥125 mg kg−1 for the 3 soils tested. These results can be considered relevant when anticipating possible environmental repercussions related to the simultaneous presence of various types of pollutants, specifically certain heavy metals and antibiotics., This study has been funded by the Spanish Ministry of Economy and Competitiveness through the projects CGL 2015-67333-C2-1-R and -2-R (FEDER Funds) and by Xunta de Galicia via BV1 research group (ED431C 2017/62-GRC). F-CD holds a Ramón y Cajal contract (RYC-2016-20411) financed by the Spanish Ministry of Economy, Industry and Competitiveness. S-MV Santás Miguel holds a posdoctoral fellowship (ED481B-2022-081) financed by Xunta de Galicia. A-RL hold a pre-doctoral fellowship (ED481A 2021/309) financed by Xunta de Galicia.

The increase of concentrations of tetracycline antibiotics in agricultural soils worldwide is of special concern, due to its potential toxic effects on soil bacterial communities. In the present work, the reuse of two waste/by-product materials as soil amendments was tested as a preventive practice for reducing tetracycline antibiotics toxicity in soils. Pine bark (PB), with high percentage of organic carbon, and crushed mussel shell (CMS), a frequent natural liming material, were added to 4 soils in doses 0, 6, 12 and 48 g of by-product per kg−1 of soil (dry weight) of each one (separately). The soils and soil-waste mixtures were then spiked with tetracycline (TC), oxytetracycline (OTC) and chlortetracycline (CTC). After one day of incubation, the bacterial growth was estimated in soils and soil-mixtures using the leucine incorporation technique. The addition of PB to the soils showed two different behaviors, depending on the antibiotics. The toxicity of TC and OTC decreased with the addition of PB (toxicities going from 6 to 25% and from 5 to 36%, respectively). However, CTC toxicity did not change, or even increased in response to the PB amendment. Regarding soil amendment with CMS, it was not effective to prevent the toxicity of any of the three antibiotics studied., This study has been funded by the Spanish Ministry of Economy and Competitiveness through the projects CGL 2015-67333-C2-1-R and -2-R (FEDER Funds), and by Xunta de Galicia CITACA Strategic Partnership (ED431E 2018/07) and BV1 research group (ED431C 2017/62-GRC). David Fernández Calviño holds a Ramón y Cajal contract (RYC-2016-20411) financed by the Spanish Ministry of Economy, Industry and Competitiveness. Vanesa Santás Miguel holds a pre-doctoral fellowship founded by the University of Vigo.