Capture and Characterization of Self-Transmissible Plasmids from Urban Wetlands E

城市湿地 E 中自传播质粒的捕获和表征

• 批准号: 8495716
• 负责人: DAVID Edward CUMMINGS
• 金额: $ 25.25万
• 依托单位: POINT LOMA NAZARENE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-01 至 2016-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8495716
• 关键词: Address; Alleles; Amino Acid Sequence; Antibiotic Resistance; Antibiotics; Bacteria; Base Sequence; Bioinformatics; Biological Assay; California; Characteristics; Clinic; Clinical; Coin; Collection; Communities; County; DNA; DNA Insertion Elements; DNA Sequence; Databases; Diffusion; Drug resistance; Environment; Epidemic; Event; Fluoroquinolones; Future; Genbank; Gene Combinations; Genes; Genetic; Genetic Recombination; Genetic Variation; Goals; Health; Human; Idaho; Indigenous; Infection; Institutes; Integrons; International; Knowledge; Lactams; Lead; Maps; Measures; Mediating; Methods; Minimum Inhibitory Concentration measurement; Mobile Genetic Elements; Monobactams; Mosaicism; Multi-Drug Resistance; Nature; Open Reading Frames; Partner in relationship; Peptide Sequence Determination; Phenotype; Plasmids; Play; Public Health; Replicon; Resistance; Rivers; Role; Techniques; Technology; Testing; Time; Universities; Vertebral column; Work; anthropogenesis; bacterial resistance; base; clinically relevant; combat; comparative genomics; fluoroquinolone resistance; genome sequencing; inhibitor/antagonist; metagenome; novel; pathogen; pollutant; prevent; public health relevance; pyrosequencing; quinolone resistance; success; tool

DESCRIPTION (provided by applicant): Urban wetlands receive contaminated runoff from all activities within the watershed and pose a potential threat to act as reservoirs of various anthropogenic pollutants. Recent work indicates that antibiotic-resistant bacteria and resistance genes wash into urban wetlands during runoff events where they accumulate and persist long after runoff subsides. The overarching goal of this project is to understand the threat posed by urban wetlands on antibiotic resistance in the clinic. Antibiotic resistance plasmids will be captured from the wetlands, and characterized to determine how likely they are spread to other species and what resistances they confer. This will be done by transferring plasmids from bacteria in the indigenous bacterial communities into a plasmid-free recipient strain by conjugation, using either a bi-parental or tri-parental mating approach, which does not require cultivation of the native plasmid hosts. Antibiotic resistances conferred by the isolated plasmids will be determined using standard disk diffusion methods. The potential for the plasmids to be transmitted to other species will be inferred from the replicon type (determined by PCR) as well as experimental host range assays using a diverse collection of potential recipient strains. The complete DNA sequences of broad host-range plasmids conferring resistance to fluoroquinolone and/or ¿-lactam antibiotics will be determined using 454 pyrosequencing technology and thoroughly annotated and mapped to determine their genetic organization. This project will reveal many things about the urban wetland plasmid metagenome including (1) collective antibiotic resistance, (2) novel resistance genes, alleles, and combinations, and (3) threat of mobilization into clinical pathogens.

描述（由申请人提供）：城市湿地接收流域内所有活动的污染径流，并构成潜在的威胁，成为各种人为污染物的水库。最近的研究表明，在径流事件期间，抗寄生虫细菌和抗性基因冲入城市湿地，在径流消退后，它们在那里积累并持续很长时间。该项目的总体目标是了解城市湿地对临床抗生素耐药性构成的威胁。将从湿地中捕获抗生素耐药性质粒，并对其进行表征，以确定它们传播到其他物种的可能性以及它们赋予的耐药性。这将通过使用双亲或三亲交配方法，通过接合将质粒从本土细菌群落中的细菌转移到无质粒的受体菌株中来完成，该方法不需要培养天然质粒宿主。由分离的质粒赋予的抗生素抗性将使用标准纸片扩散法测定。将根据复制子类型（通过PCR确定）以及使用不同的潜在受体菌株集合进行的实验宿主范围测定来推断质粒传播至其他物种的可能性。将使用454焦磷酸测序技术确定赋予氟喹诺酮和/或内酰胺类抗生素抗性的广泛宿主范围质粒的完整DNA序列，并进行彻底注释和作图，以确定其遗传组织。该项目将揭示关于城市湿地质粒宏基因组的许多事情，包括（1）集体抗生素耐药性，（2）新的耐药基因，等位基因和组合，以及（3）动员到临床病原体的威胁。