Hallewell J, Niu Y D, Munns K, McAllister T A, Johnson R P, Ackermann H-W, Thomas J E, Stanford K
Department of Biological Sciences University of Lethbridge, Lethbridge, Alberta, Canada Agriculture and Agri-Food Canada, Lethbridge, Alberta, Canada.
Alberta Agriculture and Rural Development, Lethbridge, Alberta, Canada.
Appl Environ Microbiol. 2014 Jul;80(13):3819-25. doi: 10.1128/AEM.00708-14. Epub 2014 Apr 18.
The objectives of this study were to identify endemic bacteriophages (phages) in the feedlot environment and determine relationships of these phages to Escherichia coli O157:H7 from cattle shedding high and low numbers of naturally occurring E. coli O157:H7. Angus crossbred steers were purchased from a southern Alberta (Canada) feedlot where cattle excreting ≥ 10(4) CFU · g(-1) of E. coli O157:H7 in feces at a single time point were identified as supershedders (SS; n = 6), and cattle excreting <10(4) CFU · g(-1) of feces were identified as low shedders (LS; n = 5). Fecal pats or fecal grabs were collected daily from individual cattle for 5 weeks. E. coli O157:H7 in feces was detected by immunomagnetic separation and enumerated by direct plating, and phages were isolated using short- and overnight-enrichment methods. The total prevalence of E. coli O157:H7 isolated from feces was 14.4% and did not differ between LS and SS (P = 0.972). The total prevalence of phages was higher in the LS group (20.9%) than in the SS group (8.3%; P = 0.01). Based on genome size estimated by pulsed-field gel electrophoresis and morphology determined by transmission electron microscopy, T4- and O1-like phages of Myoviridae and T1-like phage of Siphoviridae were isolated. Compared to T1- and O1-like phages, T4-like phages exhibited a broad host range and strong lytic capability when targeting E. coli O157:H7. Moreover, the T4-like phages were more frequently isolated from feces of LS than SS, suggesting that endemic phages may impact the shedding dynamics of E. coli O157:H7 in cattle.
本研究的目的是鉴定饲养场环境中的地方性噬菌体,并确定这些噬菌体与来自自然排出高数量和低数量大肠杆菌O157:H7的牛的大肠杆菌O157:H7之间的关系。从加拿大艾伯塔省南部的一个饲养场购买了安格斯杂交阉牛,在该饲养场,在单个时间点粪便中排出≥10⁴ CFU·g⁻¹大肠杆菌O157:H7的牛被鉴定为超级排菌者(SS;n = 6),而粪便中排出<10⁴ CFU·g⁻¹的牛被鉴定为低排菌者(LS;n = 5)。连续5周每天从每头牛采集粪便块或粪便样本。通过免疫磁珠分离法检测粪便中的大肠杆菌O157:H7,并通过直接平板计数法进行计数,使用短期和过夜富集方法分离噬菌体。从粪便中分离出的大肠杆菌O157:H7的总流行率为14.4%,LS组和SS组之间无差异(P = 0.972)。噬菌体的总流行率在LS组(20.9%)高于SS组(8.3%;P = 0.01)。基于脉冲场凝胶电泳估计的基因组大小和透射电子显微镜确定的形态,分离出了肌尾噬菌体科的T4-和O1-样噬菌体以及长尾噬菌体科的T1-样噬菌体。与T1-和O1-样噬菌体相比,T4-样噬菌体在靶向大肠杆菌O157:H7时表现出广泛的宿主范围和强大的裂解能力。此外,T4-样噬菌体在LS组粪便中的分离频率高于SS组,这表明地方性噬菌体可能影响牛体内大肠杆菌O157:H7的排菌动态。