Sanad Yasser M, Johnson Kelly, Park Si Hong, Han Jing, Deck Joanna, Foley Steven L, Kenney Brett, Ricke Steven, Nayak Rajesh
1 Division of Microbiology, National Center for Toxicological Research , U.S. Food and Drug Administration, Jefferson, Arkansas.
2 Center for Food Safety and Department of Food Science, University of Arkansas , Fayetteville, Arkansas.
Foodborne Pathog Dis. 2016 Feb;13(2):80-7. doi: 10.1089/fpd.2015.2002. Epub 2015 Dec 14.
This study evaluated antimicrobial resistance and virulence factors in Salmonella enterica isolated from a turkey flock in which the birds were raised in an environment where antimicrobials were not administered to the birds, either through feed or water. Salmonella was isolated from turkeys and various environmental samples in the facility using conventional microbiological procedures. Isolates were serotyped and analyzed phenotypically by antimicrobial resistance profiling and genotypically by pulsed-field gel electrophoresis (PFGE) fingerprinting, integron analysis, plasmid profiling, replicon-based incompatibility (Inc) group typing, and virulence gene profiling. Ninety-five S. enterica isolates were isolated from cecal contents (n = 29), feed (n = 22), leftover feed (n = 13), litter (n = 12), drinkers (n = 10), environment (n = 8), and an insect. The following serotypes were identified: Montevideo (24%), Anatum (22%), Agona (17%), Kentucky and Worthington (12%), Senftenberg (11%), and rough phenotypes (3%). The majority of isolates (61/95; 64%) were susceptible to 12 antimicrobials tested; however, despite the absence of antimicrobials in the facility, approximately 36% of the isolates were resistant to two to five antimicrobials. Class 1 integrons were detected in 8% of the isolates. The integron sequence analysis revealed dihydrofolate reductase (dhfr) and aminoglycoside adenylyl transferase (aadA2) genes, which encode trimethoprim and streptomycin resistance, respectively. Furthermore, 71% of the isolates had at least one plasmid. There were five plasmid replicon types identified among the isolates, including IncI1, IncHI2, IncFIIA, IncB/O, and IncP, with variable prevalence among the serotypes. All 95 isolates tested polymerase chain reaction-positive for 19 virulence genes and negative for virD4 and virB4. The virulence gene profiles were similar within the isolates from the same serotype. Within particular serotypes, PFGE patterns revealed 100% similarity, even when the bacterial strains were isolated from different sources, indicating cross-colonization of sources within the turkey facility. On this antibiotic-free turkey farm, turkeys and feed appeared to be the major reservoirs of multidrug-resistant Salmonella, which harbored multiple virulence genes.
本研究评估了从一个火鸡群中分离出的肠炎沙门氏菌的抗菌药物耐药性和毒力因子。该火鸡群饲养在一个未通过饲料或饮水给禽类使用抗菌药物的环境中。使用常规微生物学方法从该设施中的火鸡和各种环境样本中分离出沙门氏菌。对分离株进行血清分型,并通过抗菌药物耐药性谱进行表型分析,通过脉冲场凝胶电泳(PFGE)指纹图谱、整合子分析、质粒图谱分析、基于复制子的不相容性(Inc)组分型和毒力基因谱进行基因型分析。从盲肠内容物(n = 29)、饲料(n = 22)、剩余饲料(n = 13)、垫料(n = 12)、饮水器(n = 10)、环境(n = 8)和一只昆虫中分离出95株肠炎沙门氏菌。鉴定出以下血清型:蒙得维的亚(24%)、阿纳托姆(22%)、阿哥纳(17%)、肯塔基和沃辛顿(12%)、森夫滕贝格(11%)以及粗糙型(3%)。大多数分离株(61/95;64%)对所测试的12种抗菌药物敏感;然而,尽管该设施中未使用抗菌药物,但约36%的分离株对两到五种抗菌药物耐药。在8%的分离株中检测到1类整合子。整合子序列分析揭示了二氢叶酸还原酶(dhfr)和氨基糖苷腺苷酸转移酶(aadA2)基因,它们分别编码对甲氧苄啶和链霉素的耐药性。此外,71%的分离株至少有一个质粒。在分离株中鉴定出五种质粒复制子类型,包括IncI1、IncHI2、IncFIIA、IncB/O和IncP,在不同血清型中的流行率各不相同。所有95株分离株对19个毒力基因的聚合酶链反应检测均为阳性,对virD4和virB4检测为阴性。同一血清型的分离株之间毒力基因谱相似。在特定血清型中,PFGE图谱显示100%的相似性,即使细菌菌株是从不同来源分离的,这表明火鸡设施内不同来源之间存在交叉定植。在这个无抗生素的火鸡养殖场,火鸡和饲料似乎是携带多种毒力基因的多重耐药性沙门氏菌的主要储存库。