Blum Shlomo E, Heller Elimelech D, Sela Shlomo, Elad Daniel, Edery Nir, Leitner Gabriel
Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food and Environment, Rehovot, Israel; National Mastitis Reference Center, Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel; Department of Bacteriology, Kimron Veterinary Institute, Bet Dagan, Israel.
Department of Animal Sciences, Robert H. Smith Faculty of Agriculture, Food and Environment, Rehovot, Israel.
PLoS One. 2015 Sep 1;10(9):e0136387. doi: 10.1371/journal.pone.0136387. eCollection 2015.
Escherichia coli is a major etiological agent of intra-mammary infections (IMI) in cows, leading to acute mastitis and causing great economic losses in dairy production worldwide. Particular strains cause persistent IMI, leading to recurrent mastitis. Virulence factors of mammary pathogenic E. coli (MPEC) involved pathogenesis of mastitis as well as those differentiating strains causing acute or persistent mastitis are largely unknown. This study aimed to identify virulence markers in MPEC through whole genome and phenome comparative analysis. MPEC strains causing acute (VL2874 and P4) or persistent (VL2732) mastitis were compared to an environmental strain (K71) and to the genomes of strains representing different E. coli pathotypes. Intra-mammary challenge in mice confirmed experimentally that the strains studied here have different pathogenic potential, and that the environmental strain K71 is non-pathogenic in the mammary gland. Analysis of whole genome sequences and predicted proteomes revealed high similarity among MPEC, whereas MPEC significantly differed from the non-mammary pathogenic strain K71, and from E. coli genomes from other pathotypes. Functional features identified in MPEC genomes and lacking in the non-mammary pathogenic strain were associated with synthesis of lipopolysaccharide and other membrane antigens, ferric-dicitrate iron acquisition and sugars metabolism. Features associated with cytotoxicity or intra-cellular survival were found specifically in the genomes of strains from severe and acute (VL2874) or persistent (VL2732) mastitis, respectively. MPEC genomes were relatively similar to strain K-12, which was subsequently shown here to be possibly pathogenic in the mammary gland. Phenome analysis showed that the persistent MPEC was the most versatile in terms of nutrients metabolized and acute MPEC the least. Among phenotypes unique to MPEC compared to the non-mammary pathogenic strain were uric acid and D-serine metabolism. This study reveals virulence factors and phenotypic characteristics of MPEC that may play a role in pathogenesis of E. coli mastitis.
大肠杆菌是奶牛乳房内感染(IMI)的主要病原体,可导致急性乳腺炎,并在全球乳制品生产中造成巨大经济损失。特定菌株会引发持续性IMI,导致乳腺炎反复发作。乳腺致病性大肠杆菌(MPEC)的毒力因子在乳腺炎发病机制以及区分引起急性或持续性乳腺炎的菌株方面很大程度上尚不清楚。本研究旨在通过全基因组和表型组比较分析来鉴定MPEC中的毒力标志物。将引起急性(VL2874和P4)或持续性(VL2732)乳腺炎的MPEC菌株与一株环境菌株(K71)以及代表不同大肠杆菌致病型的菌株基因组进行比较。小鼠乳房内攻毒实验证实,此处研究的菌株具有不同的致病潜力,且环境菌株K71在乳腺中无致病性。全基因组序列和预测蛋白质组分析表明,MPEC之间具有高度相似性,而MPEC与非乳腺致病性菌株K71以及其他致病型的大肠杆菌基因组存在显著差异。在MPEC基因组中鉴定出且在非乳腺致病性菌株中缺乏的功能特征与脂多糖和其他膜抗原的合成、柠檬酸铁铁摄取及糖类代谢有关。分别在严重和急性(VL2874)或持续性(VL2732)乳腺炎菌株的基因组中发现了与细胞毒性或细胞内存活相关的特征。MPEC基因组与K-12菌株相对相似,此处随后表明K-12菌株在乳腺中可能具有致病性。表型组分析表明,就代谢的营养物质而言,持续性MPEC最为多样,而急性MPEC最少。与非乳腺致病性菌株相比,MPEC特有的表型包括尿酸和D-丝氨酸代谢。本研究揭示了MPEC的毒力因子和表型特征,这些可能在大肠杆菌乳腺炎的发病机制中发挥作用。