Keelara Shivaramu, Thakur Siddhartha
Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, United States.
Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University, 1060 William Moore Drive, Raleigh, NC 27607, United States.
Vet Microbiol. 2014 Sep 17;173(1-2):76-83. doi: 10.1016/j.vetmic.2014.07.018. Epub 2014 Aug 1.
The aim of this study was to characterize and determine the inter-serovar exchange of AmpC β-lactamase conferring plasmids isolated from humans, pigs and the swine environment. Plasmids isolated from a total of 21 antimicrobial resistant (AMR) Salmonella isolates representing human clinical cases (n=6), pigs (n=6) and the swine farm environment (n=9) were characterized by replicon typing and restriction digestion, inter-serovar transferability by conjugation, and presence of AmpC β-lactamase enzyme encoding gene blaCMY-2 by southern hybridization. Based on replicon typing, the majority (17/21, 81%) of the plasmids belonged to the I1-Iγ Inc group and were between 70 and 103kb. The potential for inter-serovar plasmid transfer was further confirmed by the PCR detection of AMR genes on the plasmids isolated from trans-conjugants. Plasmids from Salmonella serovars Anatum, Ouakam, Johannesburg and Typhimurium isolated from the same cohort of pigs and their environment and S. Heidelberg from a single human clinical isolate had identical plasmids based on digestion with multiple restriction enzymes (EcoRI, HindIII and PstI) and southern blotting. We demonstrated likely horizontal inter-serovar exchange of plasmid-encoding AmpC β-lactamases resistance among MDR Salmonella serotypes isolated from pigs, swine farm environment and clinical human cases. This study provides valuable information on the role of the swine farm environment and by extension other livestock farm environments, as a potential reservoir of resistant bacterial strains that potentially transmit resistance determinants to livestock, in this case, swine, humans and possibly other hosts by horizontal exchange of plasmids.
本研究的目的是对从人类、猪和猪养殖环境中分离出的携带AmpC β-内酰胺酶的质粒进行特征分析,并确定其血清型间的交换情况。通过复制子分型和限制性酶切对从总共21株耐抗菌药物(AMR)的沙门氏菌分离株中分离出的质粒进行特征分析,这些分离株代表人类临床病例(n = 6)、猪(n = 6)和猪场环境(n = 9),通过接合试验检测血清型间的转移性,并通过Southern杂交检测AmpC β-内酰胺酶编码基因blaCMY-2的存在情况。基于复制子分型,大多数质粒(17/21,81%)属于I1-Iγ Inc组,大小在70至103kb之间。从转接合子中分离出的质粒上AMR基因的PCR检测进一步证实了血清型间质粒转移的可能性。从同一猪群及其环境中分离出的沙门氏菌血清型阿纳托姆、瓦卡姆、约翰内斯堡和鼠伤寒沙门氏菌以及从一名人类临床分离株中分离出的海德堡沙门氏菌的质粒,经多种限制性酶(EcoRI、HindIII和PstI)消化和Southern印迹分析后具有相同的质粒。我们证明了在从猪、猪场环境和人类临床病例中分离出的多重耐药沙门氏菌血清型之间,可能存在编码AmpC β-内酰胺酶耐药性的质粒的水平血清型间交换。本研究提供了有价值的信息,说明猪场环境以及由此延伸的其他畜牧场环境,作为耐药菌株的潜在储存库,可能通过质粒的水平交换将耐药决定因子传播给家畜,在本案例中是猪、人类以及可能的其他宿主。