Preliminary Study On The Antibiotic Resistance Gene Pollution In Fifteen Typical Lake Sediments

[Objective] Because of the extensive use of antibiotic and other antimicrobial drugs in the medicine and food animal producing industry, the antibiotics resistance has emerged and become urgent issue. The rising of the antibiotic pollution in the water environment promotes the spread of the antibiotic resistance and evolution of the antibiotic resistance genes (ARGs). However, the antibiotic resistance studies only focus on a few drugs with limited approaches, and little is known about the antibiotic resistiome in the water environment, and the effect of antibiotics on the ARGs. It is also not clear about the mechanism of transmission of ARGs between the water environment and microbiota. This study employed PCR and Q-PCR approach to investigated that lake water quality prevalence of antibiotic resistance gene, understand the effect to antibiotics on the prevalence and evolution of ARGs, and reveal the potential mechanism of transmission of ARGs. With this knowledge, we can better evaluate the impact of ARGs on the water environment.[Methods] We collected 15 domestic typical lake sediment as the research object, using indirect extraction method and direct extraction method and the obtained sediment microbial genome DNA, by ordinary PCR and fluorescence quantitative PCR amplification of resistance gene and dissemination of components of integrons and resistance gene cassettes, relative content of resistant gene detection and analysis the transmission mechanisms of resistance gene in China Lake Sediments of epidemiology and drug resistance gene.[Result] 1. The sediment microbial genomic DNA were extracted directly by protease K+SDS method, and combined with OGMEA purification kit to remove residual, humus, heavy metals and other related impurities, we can get a large number of high quality, complete the genomic DNA.2. The 28 kinds of resistance genes were detected by common PCR technology, and the resistance genes were mainly concentrated in the middle and lower reaches of the Yangtze River (Dongting Lake, Poyang Lake, Chaohu, Taihu, Yang Chenghu); Huaihe River Valley (Hong Zehu), the lower reaches of the Yellow River (Weishan Lake) and the southwest region (Dianchi). Relatively speaking, in cold plateau region (wrong Basongco lake, Qinghai Lake, Namco, Caka Salt Lake) and high latitude alpine region of Xingkai Lake resistant gene detection rate is low.3. Detection of the relative content of resistance genes by fluorescence quantitative PCR:The relative content of Yangtze River (Dongting Lake, Poyang Lake, Chaohu, Taihu, Yang Chenghu); Huaihe River Valley (Hong Zehu), the lower reaches of the Yellow River (Weishan Lake) southwestern Dianchi lake sediment sample varieties resistant gene was significantly higher than that in the alpine region (wrong Basongco lake, Qinghai Lake, Namtso, Caka Salt Lake) and high latitude alpine region in Xingkai Lake sediment samples of resistance genes relative content.4. Using ordinary PCR detection of spread components integrons and resistance gene cassettes in resistance genes, sediment microbial genome DNA extracted by direct method for template detection ?, ?, ? class integron, the detection rate was 80%,100%,46.67%. And the detection to the class 1 integron a aada2 gene.[Conclusion] This experiment of 14 kinds of tetracycline resistance genes,12 kinds of beta-lactamase resistance genes and 2 kinds of aminoglycoside resistance gene study found that water environment antibiotic resistance genes mainly concentrated in the middle and lower reaches of the Yangtze River (Dongting Lake, Poyang Lake, Chaohu, Taihu, Yang Chenghu); Huaihe River Valley (Hong Zehu), the lower reaches of the Yellow River (Weishan Lake) and the southwest region (Dianchi). Relatively speaking, the relative content of antibiotic resistance genes in cold plateau region (wrong Basongco lake, Qinghai Lake, Namco, Caka Salt Lake) and high latitude alpine region of Xingkai Lake is lower.