Biogas slurry and residue contaminated with antibiotics are widely used as fertilizers in vegetable crop planting. However, their impact on the spreading of antibiotic resistance genes (ARGs) in vegetable fields is still largely unknown. In the present study, antibiotic resistant bacteria (ARB), ARGs and bacterial communities from pig manure to fields were monitored by using viable plate counts, high-throughput fluorescent quantitative PCR (HT-qPCR) and Illumina MiSeq sequencing. Eighty-three ARGs and 3 transposons genes were detected. Anaerobic digestion reduced relative abundance of tetracycline and Macrolide-Lincosamide-Streptogramin (MLSB) resistance genes. However, the number of ARB and the relative abundance of sulfa, aminoglycoside and florfenicol, chloramphenicol, and amphenicol (FCA) resistance genes, respectively, enriched up to 270 times and 52 times in biogas residue. Long-term application of biogas slurry and residue contaminated with antibiotics in fields increased the rate of ARB as well as relative abundance of ARGs and transposons genes. Additionally, bacterial communities significantly differed between the soil treated with biogas slurry and residue and the control sample, especially the phyla Bacteroidetes and Actinobacteria. Based on network analysis, 19 genera were identified as possible hosts of the detected ARGs. Our results provide an important significance for reasonable application of biogas slurry and residue.

中文翻译：

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沼液和残渣直接施用在田间的影响：从猪粪到田间的抗生素抗性基因的原位分析

沼气浆液和被抗生素污染的残留物被广泛用作蔬菜作物种植中的肥料。然而，其对蔬菜田中抗生素抗性基因（ARGs）传播的影响仍是未知之数。在本研究中，通过使用活板计数，高通量荧光定量PCR（HT-qPCR）和Illumina MiSeq测序监测从猪粪到田间的抗生素抗性细菌（ARB），ARG和细菌群落。共检测到83个ARGs和3个转座子基因。厌氧消化降低了四环素和大环内酯-林可酰胺-链霉菌素（MLSB）抗性基因的相对丰度。但是，分别具有抗性的ARB数量和磺胺类，氨基糖苷类和氟苯尼考，氯霉素和苯酚（FCA）抗性基因的相对丰度，富集了270倍和52倍的沼气残渣。在田间长期使用沼液和被抗生素污染的残留物可以增加ARB的发生率以及ARG和转座子基因的相对丰度。此外，用沼气泥浆和残渣处理过的土壤与对照样品（尤其是门）之间的细菌群落也存在显着差异。拟杆菌和放线菌。根据网络分析，确定了19个属是检测到的ARG的可能宿主。我们的结果为合理利用沼液和残渣提供了重要的意义。