摘要
出身背景
抗生素耐药性已被公认为全球关注的一种现象，是由于细菌对抗菌药物的广泛使用而产生的反应。抗性决定簇通过水平转移的扩展与移动遗传元件（MGE）有关，如转座子、插入序列和整合子。重金属也会对健康造成危害。金属抗性基因与抗生素抗性基因（ARGs）和MGEs联合，帮助细菌获得更高的抗性。
方法论
本工作通过对克什米尔的Wular湖和Dal湖进行PCR和测序来研究ARGs blaCTX-M、AmpC、qnrS、MGE如ISecp1、TN3、TN21和Int I；印度使用PCR扩增和测序方法进行blaCTX-M-15的遗传环境分析，然后进行计算机对接和突变研究。确定了ARGs和HMRGs的共同出现。使用基于PCR的复制子分型（PBRT）进行质粒分型，并进行缀合测定。
后果
在从16个地点获得的201个分离株中，33个是ESBLs的生产者。30个显示ESBL的分离株被认为对CTX-M基因呈阳性，其次是AmpC（17）、qnrS（13）、ISecp1（15）、TN3（11）、TN21（11），Int I（18）和SulI（14）。blaCTX-M-15的遗传环境为（ISEcp1-laCTX-M-15-orf477），在3′区发现经典启动子-10 TACAAT和-35 TTGAA。产生了CTX-M-15和ISEcp1的3D结构，并且CTX-M-15-ISEcp1（R299L）对接和突变显示氢键减少。在18、14和8个分离株中发现了抗生素和HMRGs（mer、sil和ars）的共存。PBRT分析显示存在股份有限公司组-B/O、F、I1、HI1、FIA、HI2、N、FIB、L/M。转导偶联物的分子分析表明，ARGs、MGEs和HMRGs在大肠杆菌J53-AZR菌株中成功转移。
结论
这项研究强调了产ESBL的细菌在印度克什米尔的水生环境中的存在，这些细菌可以作为ARGs的蓄水池。它还讨论了MGEs的分子机制，这有助于遏制抗生素耐药性的传播。
Abstract
Background
Antibiotic resistance conceded as a global concern is a phenomenon that emerged from the bacterial response to the extensive utilization of antimicrobials. The expansion of resistance determinants through horizontal transfer is linked with mobile genetic elements (MGEs) like transposons, insertion sequences, and integrons. Heavy metals also create consequential health hazards. Metal resistance gene in alliance with antibiotic resistance genes (ARGs) and MGEs is assisting bacteria to attain exalted quantity of resistance.

Methodology
The present work was carried out to study ARGs blaCTX-M, AmpC, qnrS, MGEs like ISecp1, TN3, TN21, and Int I by performing PCR and sequencing from Wular and Dal lakes of Kashmir; India. The genetic environment analysis of blaCTX-M-15 was carried out using PCR amplification, and sequencing approach followed by in-silico docking and mutational studies. Co-occurrence of ARGs and HMRGs was determined. Plasmid typing was done using PCR-based replicon typing (PBRT) and conjugation assay was also performed.

Results
Out of 201 isolates attained from 16 locations, 33 were ESBLs producers. 30 ESBL displaying isolates were perceived positive for CTX-M gene, followed by AmpC (17), qnrS (13), ISecp1 (15), TN3 (11), TN21 (11), Int I (18), and SulI (14). The genetic environment of blaCTX-M-15 was observed as (ISEcp1-blaCTX-M-15-orf477), classical promoter-10 TACAAT and -35 TTGAA was found at the 3′ region. The 3D structure of CTX-M-15 and ISEcp1 was generated and CTX-M-15-ISEcp1 (R299L) docking and mutation showed a reduction in hydrogen bonds. Co-occurrence of antibiotics and HMRGs (mer, sil, and ars) was found in 18, 14, and 8 isolates. PBRT analysis showed the presence of Inc. groups- B/O, F, I1, HI1, FIA, HI2, N, FIB, L/M. Molecular analysis of transconjugants showed the successful transfer of ARGs, MGEs, and HMRGs in the E. coli J53 AZR strain.

Conclusion
This study highlights the occurrence of ESBL producing bacteria in the aquatic environment of Kashmir India that can serve as a reservoir of ARGs. It also discussed the molecular mechanisms of MGEs which can help in containing the spread of antibiotic resistance.

https://www.sciencedirect.com/science/article/abs/pii/S0048969722013134