摘要

       沼渣和有机肥常用于作物种植。然而，在将其应用于田间之前，对抗生素抗性细菌（ARB）、抗生素抗性基因（ARGs）和细菌群落的评估仍然缺乏。本研究通过使用活菌落数、高水平的微生物菌落数，监测了对磺胺嘧啶、四环素和诺氟沙星耐药的细菌、对磺胺类、四环素和氟喹诺酮类的 57 个耐药基因的变异以及在 28 天有氧储存沼渣和有机肥期间的细菌群落。 - 通量 qPCR 和 Illumina MiSeq 测序方法。然后使用孵化器（25°C）和自然环境这两种储存条件来评估 ARB 和 ARGs 对环境因素的反应。结果表明，在沼渣和有机肥中分别检测到 35 和 21 种 ARGs。 ARB 和 ARGs 在储存 28 天后沼渣中的富集高达 8.01 倍，但在有机肥储存期间变化范围很窄。与培养箱条件相比，自然条件下沼渣中ARB和ARGs的增殖受到多变复杂环境因素的相对抑制。然而，我们发现孵化器和自然条件下有机肥中ARB和ARGs没有显着差异。在沼渣储存过程中，细菌群落也发生了变化，尤其是拟杆菌_VC2.1_Bac22、水母藻、黄体单胞菌和Arenimonas。网络分析表明，沼渣中的关系比有机肥中的复杂得多，最终导致沼渣短期储存过程中大量的ARB和ARGs。因此，我们建议沼渣在田间应用前应采取进一步措施。

       Biogas residue and organic manure are frequently used for crop planting. However, the evaluation of antibiotic resistant bacteria (ARB), antibiotic resistance genes (ARGs) and bacterial community before their applications to fields is still lacking. This study monitored the variations of bacteria resistant to sulfadiazine, tetracycline and norfloxacin, 57 resistance genes for sulfonamides, tetracyclines and fluoroquinolones as well as the bacterial community during the 28-day aerobic storage of biogas residue and organic manure by using viable plate counts, high-throughput qPCR and Illumina MiSeq sequencing methods. Then two storage conditions, incubator (25 °C) and natural environment, were used to assess the responses of ARB and ARGs to the environmental factors. Results showed that a total of 35 and 21 ARGs were detected in biogas residue and organic manure, respectively. ARB and ARGs were enriched up to 8.01-fold in biogas residue after the 28-day storage, but varied in a narrow range during the storage of organic manure. Compared with the incubator condition, the proliferation of ARB and ARGs in biogas residue under the natural condition was relatively inhibited by the varied and complicated environmental factors. However, we found that there was no significant difference of ARB and ARGs in organic manure between the incubator and natural conditions. Bacterial community was also shifted during the storage of biogas residue, especially Bacteroidetes_VC2.1_Bac22, Aequorivita, Luteimonas and Arenimonas. Network analysis revealed that the relationship in biogas residue was much more complicated than that in organic manure, which ultimately resulted in large successions of ARB and ARGs during the short-term storage of biogas residue. Therefore, we suggest that further measures should be taken before the application of biogas residue to fields.

https://www.sciencedirect.com/science/article/abs/pii/S0269749118324783