Grevskott Didrik H, Svanevik Cecilie S, Sunde Marianne, Wester Astrid L, Lunestad Bjørn T
National Institute of Nutrition and Seafood Research Bergen, Norway.
Norwegian Veterinary InstituteOslo, Norway; Norwegian Institute of Public HealthOslo, Norway.
Front Microbiol. 2017 Jan 18;8:24. doi: 10.3389/fmicb.2017.00024. eCollection 2017.
The mechanisms for the development and spread of antibacterial resistance (ABR) in bacteria residing in environmental compartments, including the marine environment, are far from understood. The objective of this study was to examine the ABR rates in and other Enterobacteriaceae isolates obtained from marine bivalve mollusks collected along the Norwegian coast during a period from October 2014 to November 2015. A total of 549 bivalve samples were examined by a five times three tube most probable number method for enumeration of in bivalves resulting in 199 isolates from the positive samples. These isolates were identified by biochemical reactions and matrix Assisted Laser Desorption Ionization-Time of Flight Mass Spectrometry, showing that 90% were , while the remaining were species within the genera , and . All 199 isolates recovered were susceptibility tested following the European Committee on Antimicrobial Susceptibility Testing disk diffusion method. In total, 75 of 199 (38%) isolates showed resistance to at least one antibacterial agent, while multidrug-resistance were seen in 9 (5%) isolates. One isolate conferred resistance toward 15 antibacterial agents. Among the 75 resistant isolates, resistance toward extended-spectrum penicillins (83%), aminoglycosides (16%), trimethoprim (13%), sulfonamides (11%), tetracyclines (8%), third-generation cephalosporins (7%), amphenicols (5%), nitrofurans (5%), and quinolones (5%), were observed. Whole-genome sequencing on a selection of 10 isolates identified the genes responsible for resistance, including genes. To indicate the potential for horizontal gene transfer, conjugation experiments were performed on the same selected isolates. Conjugative transfer of resistance was observed for six of the 10 isolates. In order to compare isolates from bivalves with clinical strains, multiple-locus variable number tandem repeats analysis (MLVA) was applied on a selection of 30 resistant isolates. The MLVA-profiles were associated with community-acquired strains causing bacteremia. Our study indicates that bivalves represent an important tool for monitoring antibacterial resistant and other members of the Enterobacteriaceae family in the coastal environment.
环境隔室(包括海洋环境)中细菌的抗菌耐药性(ABR)产生和传播机制仍远未明确。本研究的目的是检测2014年10月至2015年11月期间从挪威海岸采集的海洋双壳贝类软体动物中大肠杆菌及其他肠杆菌科分离株的ABR发生率。采用五次三管最大可能数法对总共549份双壳贝类样本进行检测,以计数双壳贝类中的大肠杆菌,从阳性样本中获得了199株分离株。通过生化反应和基质辅助激光解吸电离飞行时间质谱对这些分离株进行鉴定,结果显示90%为大肠杆菌,其余为克雷伯菌属、肠杆菌属和沙雷菌属中的菌种。按照欧洲抗菌药物敏感性试验委员会纸片扩散法对所有199株回收的分离株进行药敏试验。总共199株中有75株(38%)对至少一种抗菌药物耐药,9株(5%)分离株表现出多重耐药。一株分离株对15种抗菌药物耐药。在75株耐药分离株中,观察到对超广谱青霉素(83%)、氨基糖苷类(16%)、甲氧苄啶(13%)、磺胺类(11%)、四环素类(8%)、第三代头孢菌素(7%)、氯霉素类(5%)、硝基呋喃类(5%)和喹诺酮类(5%)耐药。对10株大肠杆菌分离株进行全基因组测序,确定了耐药相关基因,包括bla基因。为了表明水平基因转移的可能性,对相同的选定分离株进行了接合试验。在10株大肠杆菌分离株中有6株观察到耐药性的接合转移。为了将双壳贝类中的大肠杆菌分离株与临床菌株进行比较,对30株耐药大肠杆菌分离株进行多位点可变数目串联重复序列分析(MLVA)。MLVA图谱与引起菌血症的社区获得性大肠杆菌菌株相关。我们的研究表明,双壳贝类是监测沿海环境中抗菌耐药大肠杆菌及其他肠杆菌科成员的重要工具。