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定量和表征东亚和北美的 15 家污水处理厂中的β-内酰胺类抗生素耐药基因。

Quantification and characterization of β-lactam resistance genes in 15 sewage treatment plants from East Asia and North America.

机构信息

Environmental Biotechnology Lab, Department of Civil Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong SAR, China.

出版信息

Appl Microbiol Biotechnol. 2012 Sep;95(5):1351-8. doi: 10.1007/s00253-011-3810-5. Epub 2011 Dec 28.

Abstract

The emerging antibiotic resistance genes in the aquatic environment have aroused public concern. As β-lactam is the most widely used group of antibiotics, β-lactam resistance genes were selected to investigate their distribution and diversity in the activated sludge from 15 geographically different sewage treatment plants (STPs) of China, Singapore, USA, and Canada. Specific PCR and quantitative real-time PCR (q-PCR) were used to investigate the occurrence and abundance of nine β-lactam resistance genes. Five genes (OXA-1, OXA-2, OXA-10, ampC, and TEM-1) were detected in most of the sludge collected, while three genes (mecA, CTX-M-1, and SME) were not found in any sludge sample. The total abundances of the six detected β-lactam resistance genes in the 15 STPs varied from 5.34 × 10(1) copies/ng DNA (ampC) to 5.49 × 10(4) copies/ng DNA (OXA-1). Overall, OXA-1 had the highest total concentration, followed by IMP and OXA-10. Noticeably, the abundances of TEM-1 in Chinese STPs were generally higher than those in the STPs of other countries, while the abundances of OXA-2 and IMP in the STPs of North America were much greater than those of East Asia. A total of 78 clones carrying β-lactam resistance genes were randomly selected from six clone libraries for phylogenetic diversity analysis; the similarity of these cloned genes to known β-lactam resistance genes with sequence identities ranged from 96% to 100%. Furthermore, OXA-1, ampC, and IMP were found to be more diverse than the other β-lactam resistance genes.

摘要

水生环境中新兴的抗生素耐药基因引起了公众关注。由于β-内酰胺类抗生素是应用最广泛的抗生素之一,因此选择β-内酰胺类耐药基因来研究它们在中国、新加坡、美国和加拿大的 15 个地理位置不同的污水处理厂(STP)的活性污泥中的分布和多样性。采用特异性 PCR 和实时定量 PCR(q-PCR)技术来研究 9 种β-内酰胺类耐药基因的发生和丰度。在大多数采集的污泥中检测到了 5 种基因(OXA-1、OXA-2、OXA-10、ampC 和 TEM-1),而在任何污泥样本中都未检测到 3 种基因(mecA、CTX-M-1 和 SME)。在这 15 个 STP 中,所检测到的 6 种β-内酰胺类耐药基因的总丰度范围从 5.34×10(1)拷贝/ng DNA(ampC)到 5.49×10(4)拷贝/ng DNA(OXA-1)。总体而言,OXA-1 的总浓度最高,其次是 IMP 和 OXA-10。值得注意的是,中国 STP 中 TEM-1 的丰度通常高于其他国家的 STP,而北美的 OXA-2 和 IMP 的丰度则远高于东亚。从六个克隆文库中随机选择了 78 个携带β-内酰胺类耐药基因的克隆进行系统发育多样性分析;这些克隆基因与具有序列同一性的已知β-内酰胺类耐药基因的相似性范围为 96%至 100%。此外,OXA-1、ampC 和 IMP 比其他β-内酰胺类耐药基因具有更高的多样性。

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