Department of Biology, University of Regina, Regina, SK, Canada.
J Appl Microbiol. 2012 Jun;112(6):1123-33. doi: 10.1111/j.1365-2672.2012.05301.x. Epub 2012 Apr 26.
To isolate and characterize multiple antibiotic resistance plasmids found in swine manure and test for plasmid-associated genetic markers in soil following manure application to an agricultural field.
Plasmids were isolated from an erythromycin enrichment culture that used liquid swine manure as an inoculant. Plasmids were transformed into Escherichia coli DH10β for subsequent characterization. We isolated and DNA sequenced a 22 102-bp plasmid (pMC2) that confers macrolide, and tetracycline resistances, and carries genes predicted to code for mercury and chromium resistance. Conjugation experiments using an pRP4 derivative as a helper plasmid confirm that pMC2 has a functional mobilization unit. PCR was used to detect genetic elements found on pMC2 in DNA extracted from manure amended soil.
The pMC2 plasmid has a tetracycline-resistant core and has acquired additional resistance genes by insertion of an accessory region (12 762 bp) containing macrolide, mercury and chromium resistance genes, which was inserted between the truncated DDE motifs within the Tn903/IS102 mobile element.
Liquid swine manure used for manure spreading contains multiple antibiotic resistance plasmids that can be detected in soil following manure application.
从猪粪中分离和鉴定多种抗生素耐药质粒，并在将猪粪施用于农田后检测土壤中与质粒相关的遗传标记。
从以液态猪粪为接种物的红霉素富集培养物中分离质粒。将质粒转化为大肠杆菌 DH10β，以进行后续鉴定。我们分离并对 22102bp 的质粒（pMC2）进行了 DNA 测序，该质粒赋予了大环内酯类和四环素类耐药性，并携带了预测编码汞和铬抗性的基因。使用 pRP4 衍生物作为辅助质粒的接合实验证实了 pMC2 具有功能性的移动单元。PCR 用于检测从添加粪便的土壤中提取的 DNA 中 pMC2 上的遗传元件。
pMC2 质粒具有四环素耐药核心，并通过插入一个包含大环内酯类、汞和铬抗性基因的附加区域（12762bp）获得了额外的抗性基因，该区域插入到 Tn903/IS102 移动元件内截断的 DDE 基序之间。
用于粪肥散布的液态猪粪中含有多种抗生素耐药质粒，在粪肥施用于农田后可在土壤中检测到。
