翻堆频次对奶牛粪污异位发酵床中抗生素抗性基因的影响
作者:
作者单位:

1.武汉市农业科学院畜牧兽医研究所,武汉 430208;2.华中农业大学动物医学院,武汉 430070;3.长江大学动物科学学院,荆州 434023

作者简介:

崔华威,E-mail:29908956@qq.com

通讯作者:

金尔光,E-mail:523229599@qq.com

中图分类号:

X713

基金项目:

武汉市农业科学院协同创新专项(XTCX2020002; XTCX2021002)


Effects of stacking frequency on antibiotic resistance genes in ectopic fermentation bed for manure and sewage of dairy cows
Author:
Affiliation:

1.Institute of Animal Husbandry and Veterinary Medicine,Wuhan Academy of Agricultural Sciences,Wuhan 430208,China;2.College of Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China;3.College of Animal Science and Technology,Yangtze University,Jingzhou 434023,China

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    摘要:

    为了探讨翻堆次数对异位发酵床连续堆肥过程中抗生素抗性基因(ARGs)和可移动基因元件(MGEs)的影响,设置1 d翻堆1次(F1组)和2 d翻堆1次(F2组)2种条件,采用PCR技术检测堆肥过程中15种基因,包括2种四环素类(tetGtetW)、3种磺胺类(sul1sul2dfrA1)、2种β-内酰胺类(blaTEM-1fexA)、2种MLSB类(ermXermQ)、2种FCA类(optrAIsaE)、1种氨基糖苷类(aac(6?)-Ib-cr)、1种整合子(intI1)及2种转座子(Tn916/1545ISCRI)基因。PCR结果显示,2组样品中均检出11种ARGs(tetGtetWsul1sul2blaTEM-1fexAermXermQoptrAIsaEaac(6?)-Ib-cr)和3种MGEs(intI1Tn916/1545ISCR1),其中7种基因(tetG、tetW、sul1、sul2、blaTEM-1、ermQ、intI1)的检出率较高;F1和F2组0~5 d均检出12种,33 d和40 d分别检出1种,40 d后明显增加,表明ARGs和MGEs种类随温度的变化而改变。采用qPCR技术对检出率较高的7种基因进行检测,结果显示,F1和F2组7种目标基因的总相对丰度呈先降低后升高再降低的趋势,试验结束时较0 d分别降低82.33%和78.89%,其中tetG、tetW、sul1、blaTEM-1、ermQ、intI1的相对丰度分别降低16.51%、87.89%、54.58%、99.99%、97.80%、59.29%和64.32%、99.46%、50.91%、99.29%、82.22%、99.92%。结果表明,异位发酵床降解粪污高温期可减少ARGs种类和丰度,且2 d翻堆1次对大部分ARGs的去除效果更好。

    Abstract:

    In order to investigate the effect of stacking frequency on antibiotic resistance genes (ARGs) and movable genetic elements (MGEs) in continuous composting of ectopic fermentation bed,two conditions were set: turning over once every day (F1 group) and turning over once every two days (F2 group). Fifteen target genes,including 2 tetracycline (tetGtetW),3 sulfonamide (sul1sul2dfrA1),2 β-lactamse (blaTEM-1fexA),2 MLSB (ermXermQ),2 FCA (optrAIsaE) and 1 aminoglycoside (aac(6?)-Ib-cr),1 integrase (intI1) and 2 transposon (Tn916/1545,ISCRI) genes,were identified by PCR in composting process,and 7 genes with high detection rate (tetGtetWsul1sul2blaTEM-1ermQintI1) were quantitatively detected by qPCR. The PCR results showed that,11 kinds of ARGs (tetGtetWsul1sul2blaTME-1fexAermXermQoptrAIsaEaac (6?)-Ib-cr) and 3 kinds of MGEs (intI1,Tn916/1545,ISCRI) were detected in both groups. The detection rates of 7 genes (tetGtetWsul1sul2blaTME-1,ermQ、intI1) were high. In F1 and F2 groups,12 species were detected at 0 to 5 d,and 1 species was detected at 33 d in F1 and 40 d in F2,and the detection rates increased significantly after 40 d,indicating that the species of ARGs and MGEs changed with temperature. qPCR results showed that the total relative abundance of 7 target genes in F1 and F2 groups decreased firstly,then increased and then decreased,and decreased by 82.33% and 78.89% at the end of the experiment compared with 0 d,respectively. The relative abundance of tetGtetWsul1blaTEM-1ermQ and intI1 decreased by 16.51%,87.89%,54.58%,99.99%,97.80%,59.29%,and 64.32%,99.46%,50.91%,99.29%,82.22%,99.92% in F1 and F2 groups,respectively. The results showed that the types and relative abundance of ARGs could be reduced by ectopic fermentation bed during degradation manure and sewage at high temperature,and the removal efficiency of most ARGs was better by turning over once every two days.

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崔华威,龚萍,郜道玉,周源,华娟,邵志勇,陈洁,万平民,金尔光,李自力.翻堆频次对奶牛粪污异位发酵床中抗生素抗性基因的影响[J].华中农业大学学报,2022,41(4):64-72

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  • 收稿日期:2022-03-08
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  • 在线发布日期: 2022-10-12
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