Escherichia coli (E. coli) is an important pathogen that causes diarrhea and death in piglets. In this work, whole genome sequencing of two E. coli strains (ZB-1, ZWW-1) isolated from Saba pigs. And focus on the relationship between drug resistance, pathogenic phenotype and genotype of the two strains. This study analyzed the drug susceptibility of the two strains. The LD50 values, tissue bacterial load and intestinal pathological changes in mice infected with the two strains. The differences in gene functions such as drug resistance, virulence, and unique genes between the two strains, as well as the genetic evolutionary relationship of housekeeping genes were analyzed. The results showed that the two strains had the same resistance phenotype to most drugs. The LD50 value, tissue load, and pathological changes in mice infected with strain ZB-1 revealed that this strain was more virulent and pathogenic than strain ZWW-1. In addition, the housekeeping genes contained in the two strains are in the same large branch as E. coli of different species, and the genetic evolution is stable. All of them carry EPEC-type strain-specific virulence genes escV and ent, indicating that they are all new members of EPEC-type strains. This study laid the foundation for understanding the genetic background and biological characteristics of E. coli from Saba pigs.
High pathogenicity island (HPI), which is widely distributed in Escherichia coli (E. coli), can enhance the pathogenicity of E. coli. Thus the HPI positive E. coli could pose a threat to human and animal health. It remains to be elucidated how HPI affects the virulence of pathogenic E. coli. Autophagy is an important mechanism to maintain cellular homeostasis and an innate immunity responses of organisms against pathogens. The interaction between pathogenic E. coli possessing HPI (E. coli HPI) and host autophagy system has not been reported. In this study, it was demonstrated that pathogenic E. coli induced autophagy in 3D4/21 macrophages and HPI was associated with enhanced autophagy through transmission electron microscopy, immunofluorescence and real-time PCR. The PI3K/Akt/mTOR pathway is an important negative regulatory pathway for autophagy. Through detecting the expression of key genes of PI3K/Akt/mTOR pathway, it was speculated that HPI enhanced the inhibition of the signaling pathway stimulated by pathogenic E. coli. Furthermore, HPI inhibited the secretion of IFN-γ, while the presence of HPI did not significantly affect the secretion of IL-1β. This work is the first attempt to explore the interplay between HPI carried by pathogenic E. coli and host cell autophagy. The findings might enable better understanding of the contribution of HPI to pathogenicity.
为了解撒坝猪源大肠杆菌噬菌体分离株的生物学特性.自撒坝猪源大肠杆菌中分离到一株裂解性噬菌体,利用PCR扩增噬菌体衣壳蛋白gp23基因,并通过同源性比对及遗传进化分析来鉴定噬菌体.经增殖纯化培养后测定噬菌体的效价和对菌株的裂解效应,而后测定其最佳感染复数、热稳定性、pH稳定性及绘制生长曲线以探究该噬菌体的生物学特性.结果 表明,分离株衣壳蛋白gp23基因与噬菌体slur14亲缘关系最近,属于有尾噬菌体目、肌尾噬菌体科T4-like噬菌体;分离株具有较高的效价,达8.67×109 pfu/m;针对实验室保存的92株已确定血清型的大肠杆菌裂解率为61.95%,完全裂解率为27.17%;在60℃条件下作用30 min,或在pH<5和pH>9时,噬菌体的效价表现明显下降;生长曲线绘制后可知该噬菌体感染宿主菌的潜伏时间约为30 min,爆发时间约为120 min,裂解量为41 pfu/cell.该试验成功分离得到裂解能力较强、裂解谱较广且对理化因素耐受力较强的T4-like噬菌体.