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首页> 美国卫生研究院文献>Frontiers in Microbiology >Transferable antibiotic resistance plasmids from biogas plant digestates often belong to the IncP-1ε subgroup
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Transferable antibiotic resistance plasmids from biogas plant digestates often belong to the IncP-1ε subgroup

机译：来自沼气厂消化物的可转移抗生素抗性质粒通常属于IncP-1ε亚组

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Manure is known to contain residues of antibiotics administered to farm animals as well as bacteria carrying antibiotic resistance genes (ARGs). These genes are often located on mobile genetic elements. In biogas plants (BGPs), organic substrates such as manure and plant material are mixed and fermented in order to provide energy, and resulting digestates are used for soil fertilization. The fate of plasmid carrying bacteria from manure during the fermentation process is unknown. The present study focused on transferable antibiotic resistance plasmids from digestates of seven BGPs, using manure as a co-substrate, and their phenotypic and genotypic characterization. Plasmids conferring resistance to either tetracycline or sulfadiazine were captured by means of exogenous plasmid isolation from digestates into Pseudomonas putida KT2442 and Escherichia coli CV601 recipients, at transfer frequencies ranging from 10-5 to 10-7. Transconjugants (n = 101) were screened by PCR-Southern blot hybridization and real-time PCR for the presence of IncP-1, IncP-1ε, IncW, IncN, IncP-7, IncP-9, LowGC, and IncQ plasmids. While 61 plasmids remained unassigned, 40 plasmids belonged to the IncP-1ε subgroup. All these IncP-1ε plasmids were shown to harbor the genes tet(A), sul1, qacEΔ1, intI1, and integron gene cassette amplicons of different size. Further analysis of 16 representative IncP-1ε plasmids showed that they conferred six different multiple antibiotic resistance patterns and their diversity seemed to be driven by the gene cassette arrays. IncP-1ε plasmids displaying similar restriction and antibiotic resistance patterns were captured from different BGPs, suggesting that they may be typical of this environment. Our study showed that BGP digestates are a potential source of transferable antibiotic resistance plasmids, and in particular the broad host range IncP-1ε plasmids might contribute to the spread of ARGs when digestates are used as fertilizer.
机译：已知粪肥含有施用于牲畜的抗生素残留以及带有抗生素抗性基因（ARG）的细菌。这些基因通常位于移动遗传元件上。在沼气厂（BGP）中，将有机基质（如肥料和植物材料）混合并发酵以提供能量，然后将产生的消化物用于土壤施肥。发酵过程中粪便中携带细菌的质粒的命运是未知的。本研究集中于使用粪便作为共底物，从七个BGP的消化物中可转移的抗生素抗性质粒，以及它们的表型和基因型特征。通过从消化物到恶臭假单胞菌KT2442和大肠杆菌CV601受体的消化物中外源质粒分离，捕获对四环素或磺胺嘧啶具有抗性的质粒，转移频率范围为10 -5 到10 -7 。通过PCR-Southern印迹杂交和实时PCR筛选转导结合体（n = 101）中是否存在IncP-1，IncP-1ε，IncW，IncN，IncP-7，IncP-9，LowGC和IncQ质粒。尽管有61个质粒尚未分配，但有40个质粒属于IncP-1ε亚组。显示所有这些IncP-1ε质粒都包含不同大小的基因tet（A），sul1，qacEΔ1，intl1和integron基因盒扩增子。对16种代表性IncP-1ε质粒的进一步分析表明，它们赋予了6种不同的多种抗生素抗性模式，其多样性似乎是由基因盒阵列驱动的。从不同的BGP捕获了显示相似限制和抗生素抗性模式的IncP-1ε质粒，这表明它们可能是这种环境的典型特征。我们的研究表明，BGP消化物是可转移的抗生素抗性质粒的潜在来源，尤其是当消化物用作肥料时，宿主范围广泛的IncP-1ε质粒可能有助于ARG的传播。

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