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Manure-based amendments influence surface-associated bacteria and markers of antibiotic resistance on radishes grown in different soil textures

Appl Environ Microbiol. 2021 Mar 12:AEM.02753-20. doi: 10.1128/AEM.02753-20. Online ahead of print.

ABSTRACT

A controlled greenhouse study was performed to determine the effect of manure or compost amendments, derived during or in absence of antibiotic treatment of beef and dairy cattle, on radish taproot-associated microbiota and indicators of antibiotic resistance when grown in different soil textures. Bacterial beta-diversity, determined by 16S rRNA gene amplicon sequencing, bifurcated according to soil texture (p<0.001, R=0.501). There was a striking cross-effect in which raw manure from antibiotic-treated and antibiotic-free beef and dairy cattle added to loamy sand (LS) elevated relative (16S rRNA gene-normalized) (by 0.9-1.9 log10) and absolute (per radish) (by 1.1-3.0 log10) abundances of intI1 (an integrase gene and indicator of mobile multi-antibiotic resistance) on radishes at harvest compared to chemical fertilizer only control conditions (p<0.001). Radishes tended to carry fewer copies of intI1 and sul1 when grown in silty clay loam than LS. Composting reduced relative abundance of intI1 on LS-grown radishes (0.6-2.4 log10 decrease vs corresponding raw manure; p<0.001). Effects of antibiotic use were rarely discernable. Heterotrophic plate count bacteria capable of growth on media containing tetracycline, vancomycin, sulfamethazine, or erythromycin tended to increase on radishes grown in turned composted antibiotic-treated dairy or beef control (no antibiotics) manures relative to the corresponding raw manure in LS (0.8-2.3 log10 increase; p<0.001), suggesting that composting sometimes enriches cultivable bacteria with phenotypic resistance. This study demonstrates that combined effects of soil texture and manure-based amendments influence the microbiota of radish surfaces and markers of antibiotic resistance illuminating future research directions for reducing agricultural sources of antibiotic resistance.ImportanceIn working towards a comprehensive strategy to combat the spread of antibiotic resistance, potential farm-to-fork routes of dissemination are gaining attention. The effects of pre-harvest factors on the microbiota and corresponding antibiotic resistance indicators on the surfaces of produce commonly eaten raw is of special interest. Here we conducted a controlled greenhouse study, using radishes as a root vegetable grown in direct contact with soil, and compared the effects of manure-based soil amendments, antibiotic use in the cattle from which the manure was sourced, composting of the manure, and soil texture, with chemical fertilizer only as a control. We noted significant effects of amendment type and soil texture on the composition of the microbiota and genes used as indicators of antibiotic resistance on radish surfaces. The findings take a step towards identifying agricultural practices that aid in reducing carriage of antibiotic resistance and corresponding risks to consumers.

PMID:33712421 | DOI:10.1128/AEM.02753-20

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