金霉素胁迫下室内粪土模型中菌群多样性与四环素类抗生素耐药基因丰度研究
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广东省自然科学基金重点项目(2016A030311029);广东省大学生科技创新训练项目(201610564119)


Microbial community diversity and tetracycline resistance gene abundance in manure-soil model under chlortetracycline stress
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    【目的】探究金霉素在粪土环境中对土壤微生物群落结构多样性和四环素类耐药基因(TRG)丰度的影响。【方法】分别设置金霉素质量0(对照)、10、100、1 000 μg作为胁迫剂量,在处理后第1、7、14、28、56天采集样品,使用高通量测序和qPCR技术研究群落结构多样性和TRG丰度的变化。【结果】厚壁菌门能耐受低剂量(10 μg)金霉素的毒害作用,对中、高剂量(100、1 000 μg)敏感;放线菌门能耐受中、高剂量的毒害作用,生存繁殖具有一定优势;变形菌门的相对丰度除第28天各剂量组均显著低于对照组外,其余时间多显著高于对照组。金霉素–猪粪–土壤模型样品中共检测出3种四环素类抗生素耐药基因(TRGs):tetOtetTtetW,3个基因丰度变化趋势相似:与第1天相比,第56天各组TRGs相对含量均显著减少;低剂量组TRGs相对含量的日消减率在Ⅱ期高于对照组和中、高剂量组,第7天TRGs相对含量显著高于对照组以及中、高剂量组;中剂量组TRGs相对含量的日消减率在Ⅳ期低于其余组,第56天TRGs相对含量显著高于其余组。【结论】在猪粪–土壤环境中,不同菌群对金霉素敏感程度不同,金霉素可改变优势菌群的相对丰度,从而引起菌群群落结构的变化。金霉素可改变TRGs相对含量的日消减率,从而影响TRGs相对含量。本研究可为下一步养殖源性抗菌药物残留的生态风险评估提供一定依据。

    Abstract:

    【Objective】 To explore the effects of chlortetracycline on microflora diversity and tetracycline resistance gene (TRG) abundance in manure-soil environment.【Method】 Four different chlortetracycline dosages were adopted, including 0 (control), 10, 100, 1 000 μg. Samples were collected on the 1st, 7th, 14th, 28th and 56th day. High-throughput sequencing and qPCR were utilized to investigate the changes of microbial community diversity and TRG abundance.【Result】 Firmicutes could tolerate toxic effect of 10 μg chlortetracycline, and was sensitive to 100 and 1 000 μg chlortetracycline. Actinobacteria could tolerate toxic effects of 100 and 1 000 μg chlortetracycline, its survival and reproduction had some advantages. The relative abundances of proteobacteria in all chlortetracycline groups on the 28th day were significantly lower than that of control group, and usually significantly higher than control group on other sampling dates. Three TRGs (tetO, tetT and tetW) were detected in chlortetracycline-manure-soil model, their changing trends of abundance were similar. Compared with the 1st day, the relative abundances in all groups on the 56th day decreased significantly. The day dissipation rates in 10 μg chlortetracycline group were significantly higher than other groups in period Ⅱ, while their relative abundances on the 7th day were significantly higher. The day dissipation rates in 100 μg chlortetracycline groups were significantly lower than other groups in period Ⅳ, while the relative abundances on the 56th day were significantly higher.【Conclusion】 In manure-soil environment, the sensitivity degrees of different bacterium communities to chlortetracycline stress were different, chlortetracycline can change microbial community composition by changing the relative abundances of dominant bacteria. Chlortetracycline stress can change the day dissipation rates of TRGs, thereby affect the relative abundances of TRGs. The results can provide a basis for further research on ecological risk evaluation of antibiotic residue in poultry raising.

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邹勇,黄蔚虹,陈永杰,冯慧君,李金红,任海鑫,孙永学.金霉素胁迫下室内粪土模型中菌群多样性与四环素类抗生素耐药基因丰度研究[J].华南农业大学学报,2018,39(5):65-73

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  • 收稿日期:2018-01-31
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  • 在线发布日期: 2018-07-18