Piggery manure used for soil fertilization is a reservoir for transferable antibiotic resistance plasmids
Piggery manure used for soil fertilization is a reservoir for transferable antibiotic resistance plasmids
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DOI：
10.1111/j.1574-6941.2008.00526.x
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发表时间：
2008-10-01
影响因子：
4.2
通讯作者：
Smalla, Kornelia
中科院分区：
文献类型：
--
作者：
Binh, Chu Thi Thanh;Heuer, Holger;Smalla, Kornelia
本研究调查了猪场粪便中可转移抗生素耐药质粒的流行情况和类型。对来自德国15个农场的粪便储存罐的样品进行了分析，这些样品代表了不同规模的牛群、肉类或仔猪生产。在GFP标记的利福平耐药大肠杆菌中捕获来自粪肥细菌的抗生素耐药质粒并进行表征。通过PCR和杂交检测，在群落总DNA中也发现了质粒类型的存在。共228 transconjugants捕获从15粪便使用选择性培养基补充阿莫西林，磺胺嘧啶或四环素。81个代表不同耐药模式或不同样品的质粒的酶切图谱可分为7类。复制子探针检测显示28个质粒属于IncN，1个属于IncW，13个属于IncP-1，19个属于最近发现的pHHV 216样质粒。44个质粒检出阿莫西林耐药基因bla-TEM，68个质粒检出磺胺类耐药基因sul 1、sul 2和/或sul 3。从群落DNA中扩增的复制子特异性序列的杂交显示，IncP-1和pHHV 216类质粒在所有粪便中检测到，而IncN和IncW的频率较低。这项研究表明，“田间规模”的猪场粪便是一个水库的广泛宿主范围的质粒赋予多种抗生素耐药基因。
In this study, the prevalence and types of transferable antibiotic resistance plasmids in piggery manure were investigated. Samples from manure storage tanks of 15 farms in Germany were analysed, representing diverse sizes of herds, meat or piglet production. Antibiotic resistance plasmids from manure bacteria were captured in gfp-tagged rifampicin-resistant Escherichia coli and characterized. The occurrence of plasmid types was also detected in total community DNA by PCR and hybridization. A total of 228 transconjugants were captured from 15 manures using selective media supplemented with amoxicillin, sulfadiazine or tetracycline. The restriction patterns of 81 plasmids representing different antibiotic resistance patterns or different samples clustered into seven groups. Replicon probing revealed that 28 of the plasmids belonged to IncN, one to IncW, 13 to IncP-1 and 19 to the recently discovered pHHV216-like plasmids. The amoxicillin resistance gene bla-TEM was detected on 44 plasmids, and sulphonamide resistance genes sul1, sul2 and/or sul3 on 68 plasmids. Hybridization of replicon-specific sequences amplified from community DNA revealed that IncP-1 and pHHV216-like plasmids were detected in all manures, while IncN and IncW ones were less frequent. This study showed that 'field-scale' piggery manure is a reservoir of broad-host range plasmids conferring multiple antibiotic resistance genes.