摘要
      从不同部门的抗微生物耐药性的全球角度来看，海鲜和海洋环境通常被认为是抗微生物耐药性基因（ARGs）和移动遗传元件（MGE）的潜在宿主；然而，关于这一领域的研究很少，结果也很少。这项研究旨在提供有关各种海鲜样本和来源中耐药性标志物含量的新数据和见解，从而在全球“一个健康”方法中对人类的潜在接触。开发并验证了一种创新的高通量qPCR筛查，以同时研究革兰氏阴性菌中41个ARGs和33个MGE的存在，包括质粒复制子、整合子和插入序列。从鳕鱼（n=24）、贝类（n=66）、扁鱼（n=53）、虾（n=10）和马鱼（n=115）的细菌菌群中分离出268个海鲜菌株，分析表明，在假单胞菌属、普罗维登西亚属、克雷伯菌属、变形杆菌属、。，和Shewanella spp.分离物以及所研究的所有细菌物种中MGE的存在。我们发现MGE的发生可能与海鲜类型以及环境、农业和收获条件有关。此外，即使在调查的一半海鲜分离株中检测到MGE，也只有12个分离株与ARG相关。结果证实了抗微生物细菌（ARB）和ARG的发生率随着与潜在粪便污染源距离的增加而降低的假设。这种独特而原始的高通量微阵列设计用于筛选革兰氏阴性菌中的ARG和MGE，可以很容易地用于监测不同环境中的抗微生物耐药性基因标记。
Abstract
From a global view of antimicrobial resistance over different sectors, seafood and the marine environment are often considered as potential reservoirs of antimicrobial resistance genes (ARGs) and mobile genetic elements (MGEs); however, there are few studies and sparse results on this sector. This study aims to provide new data and insights regarding the content of resistance markers in various seafood samples and sources, and therefore the potential exposure to humans in a global One Health approach. An innovative high throughput qPCR screening was developed and validated in order to simultaneously investigate the presence of 41 ARGs and 33 MGEs including plasmid replicons, integrons, and insertion sequences in Gram-negative bacteria. Analysis of 268 seafood isolates from the bacterial microflora of cod (n = 24), shellfish (n = 66), flat fishes (n = 53), shrimp (n = 10), and horse mackerel (n = 115) show the occurrence of sul-1, ant(3″)-Ia, aph(3′)-Ia, strA, strB, dfrA1, qnrA, and blaCTX-M-9 genes in Pseudomonas spp., Providencia spp., Klebsiella spp., Proteus spp., and Shewanella spp. isolates and the presence of MGEs in all bacterial species investigated. We found that the occurrence of MGE may be associated with the seafood type and the environmental, farming, and harvest conditions. Moreover, even if MGE were detected in half of the seafood isolates investigated, association with ARG was only identified for twelve isolates. The results corroborate the hypothesis that the incidence of antimicrobial-resistant bacteria (ARB) and ARG decreases with increasing distance from potential sources of fecal contamination. This unique and original high throughput micro-array designed for the screening of ARG and MGE in Gram-negative bacteria could be easily implementable for monitoring antimicrobial resistance gene markers in diverse contexts.

https://www.mdpi.com/2076-2607/10/6/1225