Tes from other phylogroups3,7, these phylogroups do include significant pathogens8,9. Indeed, several studies have shown that the vast majority of E. coli isolates from cases of bovine mastitis (termed mammary pathogenic E. coli, or MPEC) originate from within these two phylogroups10?4. Some lines of evidence suggest that mastitis is a general reaction to contamination of the bovine udder with any E. coli strain. For instance, the inflammatory symptoms of mastitis can be experimentally elicited by intra-mammary infusion of lipopolysaccharide (LPS) derived from cultures of E. coli of various serotypes15?7. Furthermore, some research suggests that the severity of disease is more closely dependent on factors related to the bovine host than any intrinsic differences between bacterial strains18. Previous investigations of MPEC virulence factor carriage using PCR-based screens have also been broadly unsuccessful in identifying, or agreeing upon, a core set of factors which are associated with MPEC10,13,14,19,20. However, recent evidence suggests that mastitis-causing capability in E. coli is not a general ability ?for example, Blum et al.21 showed that an environmental isolate termed E. coli K71 was incapable of causing experimental mastitis in either mice or cows21. This, along with the evidence that the molecular diversity of mastitis isolates compared with other E. coli is limited10,22, supports the conjecture that the bovine udder environment presents a milieu which is selective against the successful colonisation by some E. coli strains, yet is permissive for A-836339 chemical information others. Recently, there has been a small number of publications which have begun to examine MPEC cohorts at the genomic level21,23?6 however, few studies have examined MPEC genomes in any detail21,23. For example, Richards et al.23 compared the genome sequences of four MPEC isolates with eleven genomes of reference `commensal’ strains, such as MG1655 and HS, and principally identified a type six secretion system (T6SS), conserved in MPEC yet only sporadically present in commensal strains23. Blum et al.21 analysed the genome sequences of threeHeriot-Watt University, School of Life Sciences, Edinburgh Campus, EH14 4AS, Scotland. Correspondence and requests for materials should be addressed to D.G.E.S. (email: [email protected])received: 15 March 2016 Accepted: 27 June 2016 Published: 20 JulyScientific RepoRts | 6:30115 | DOI: 10.1038/srepwww.nature.com/scientificreports/MPEC isolates in comparison with the avirulent strain K7121. In that study, the authors identified a complement of 197 genes which were present in the genomes of MPEC, yet absent in K71. These genes included those involved in the synthesis of LPS and other membrane antigens, the capture of iron from ferric citrate, and the metabolism of certain sugars21. A recent study by Kempf et al.26 with five MPEC isolates also struggled to make progress in identifying genes implicated in the MPEC phenotype26. This study identified fifty-nine gene families which the authors postulate are MPEC-specific, however the use of somewhat relaxed inclusion criteria (presence in only two of five MPEC isolates) may reduce the likelihood that these genes impact on the MPEC phenotype. That study then used a classical candidate gene approach where they highlighted the possible role for CI-1011 site systems such as iron acquisition, fimbriae and LPS26. All of these analyses are subject to small sample size limitations, which was recognised by Richards et a.Tes from other phylogroups3,7, these phylogroups do include significant pathogens8,9. Indeed, several studies have shown that the vast majority of E. coli isolates from cases of bovine mastitis (termed mammary pathogenic E. coli, or MPEC) originate from within these two phylogroups10?4. Some lines of evidence suggest that mastitis is a general reaction to contamination of the bovine udder with any E. coli strain. For instance, the inflammatory symptoms of mastitis can be experimentally elicited by intra-mammary infusion of lipopolysaccharide (LPS) derived from cultures of E. coli of various serotypes15?7. Furthermore, some research suggests that the severity of disease is more closely dependent on factors related to the bovine host than any intrinsic differences between bacterial strains18. Previous investigations of MPEC virulence factor carriage using PCR-based screens have also been broadly unsuccessful in identifying, or agreeing upon, a core set of factors which are associated with MPEC10,13,14,19,20. However, recent evidence suggests that mastitis-causing capability in E. coli is not a general ability ?for example, Blum et al.21 showed that an environmental isolate termed E. coli K71 was incapable of causing experimental mastitis in either mice or cows21. This, along with the evidence that the molecular diversity of mastitis isolates compared with other E. coli is limited10,22, supports the conjecture that the bovine udder environment presents a milieu which is selective against the successful colonisation by some E. coli strains, yet is permissive for others. Recently, there has been a small number of publications which have begun to examine MPEC cohorts at the genomic level21,23?6 however, few studies have examined MPEC genomes in any detail21,23. For example, Richards et al.23 compared the genome sequences of four MPEC isolates with eleven genomes of reference `commensal’ strains, such as MG1655 and HS, and principally identified a type six secretion system (T6SS), conserved in MPEC yet only sporadically present in commensal strains23. Blum et al.21 analysed the genome sequences of threeHeriot-Watt University, School of Life Sciences, Edinburgh Campus, EH14 4AS, Scotland. Correspondence and requests for materials should be addressed to D.G.E.S. (email: [email protected])received: 15 March 2016 Accepted: 27 June 2016 Published: 20 JulyScientific RepoRts | 6:30115 | DOI: 10.1038/srepwww.nature.com/scientificreports/MPEC isolates in comparison with the avirulent strain K7121. In that study, the authors identified a complement of 197 genes which were present in the genomes of MPEC, yet absent in K71. These genes included those involved in the synthesis of LPS and other membrane antigens, the capture of iron from ferric citrate, and the metabolism of certain sugars21. A recent study by Kempf et al.26 with five MPEC isolates also struggled to make progress in identifying genes implicated in the MPEC phenotype26. This study identified fifty-nine gene families which the authors postulate are MPEC-specific, however the use of somewhat relaxed inclusion criteria (presence in only two of five MPEC isolates) may reduce the likelihood that these genes impact on the MPEC phenotype. That study then used a classical candidate gene approach where they highlighted the possible role for systems such as iron acquisition, fimbriae and LPS26. All of these analyses are subject to small sample size limitations, which was recognised by Richards et a.
