Identification of Shigatoxigenic and enteropathogenic Escherichia coli serotypes in healthy young dairy calves in Belgium by recto-anal mucosal swabbing

Enterohemorrhagic Escherichia coli (EHEC), enteropathogenic E. coli (EPEC), and Shigatoxigenic E. coli (STEC) are carried by healthy adult cattle and even more frequently by young calves in their intestinal tract, especially at the height of the recto-anal junction. The purpose of the present study was to assess the presence of ten EHEC, EPEC, and/or STEC O serotypes (O5, O26, O80, O103, O111, O118, O121, O145, O157, and O165) in calves sampled via recto-anal mucosal swabs (RAMS) at three dairy farms in Belgium. A total of 233 RAMS were collected on three consecutive occasions from healthy <6-month-old Holstein-Friesian calves and submitted to a PCR targeting the eae, stx1, and stx2 genes after non-selective overnight enrichment growth. The 148 RAMS testing positive were streaked on four (semi-)selective agar media; of the 2146 colonies tested, 294 from 69 RAMS were PCR-confirmed as EHEC, EPEC, or STEC. The most frequent virulotype was eae+ EPEC and the second one was stx1+ stx2+ STEC, while the eae+ stx1+ and eae+ stx1+ stx2+ virulotypes were the most frequent among EHEC. The majority of EHEC (73%) tested positive for one of the five O serotypes detected (O26, O103, O111, O145, or O157) vs. 23% of EPEC and 45% of STEC. Similarly, more RAMS (73%) harbored EHEC isolates positive for those five serotypes compared to EPEC (53%) or STEC (52%). This survey confirms that (i) healthy young dairy calves are asymptomatic carriers of EHEC and EPEC in Belgium; (ii) the carrier state rates, the virulotypes, and the identified O serotypes differ between farms and in time; and (iii) a majority of EPEC belong to so far unidentified O serotypes

Virulence of Shigatoxigenic and Enteropathogenic Escherichia coli O80:H2 in Galleria mellonella Larvae: Comparison of the Roles of the pS88 Plasmids and STX2d Phage

peer reviewedThe invasiveness properties of Shigatoxigenic and enteropathogenic Escherichia coli (STEC and EPEC) O80:H2 in humans and calves are encoded by genes located on a pS88-like ColV conjugative plasmid. The main objectives of this study in larvae of the Galleria mellonella moth were therefore to compare the virulence of eight bovine STEC and EPEC O80:H2, of two E. coli pS88 plasmid transconjugant and STX2d phage transductant K12 DH10B, of four E. coli O80:non-H2, and of the laboratory E. coli K12 DH10B strains. Thirty larvae per strain were inoculated in the last proleg with 10 μL of tenfold dilutions of each bacterial culture corresponding to 10 to 106 colony-forming units (CFUs). The larvae were kept at 37 °C and their mortality rate was followed daily for four days. The main results were that: (i) not only the STEC and EPEC O80:H2, but also different E. coli O80:non-H2 were lethal for the larvae at high concentrations (from 104 to 106 CFU) with some variation according to the strain; (ii) the Stx2d toxin and partially the pS88 plasmid were responsible for the lethality caused by the E. coli O80:H2; (iii) the virulence factors of E. coli O80:non-H2 were not identified. The general conclusions are that, although the Galleria mellonella larvae represent a useful first-line model to study the virulence of bacterial pathogens, they are more limited in identifying their actual virulence properties

Characterization of Spontaneous Bone Marrow Recovery after Sublethal Total Body Irradiation: Importance of the Osteoblastic/Adipocytic Balance

Many studies have already examined the hematopoietic recovery after irradiation but paid with very little attention to the bone marrow microenvironment. Nonetheless previous studies in a murine model of reversible radio-induced bone marrow aplasia have shown a significant increase in alkaline phosphatase activity (ALP) prior to hematopoietic regeneration. This increase in ALP activity was not due to cell proliferation but could be attributed to modifications of the properties of mesenchymal stem cells (MSC). We thus undertook a study to assess the kinetics of the evolution of MSC correlated to their hematopoietic supportive capacities in mice treated with sub lethal total body irradiation. In our study, colony-forming units – fibroblasts (CFU-Fs) assay showed a significant MSC rate increase in irradiated bone marrows. CFU-Fs colonies still possessed differentiation capacities of MSC but colonies from mice sacrificed 3 days after irradiation displayed high rates of ALP activity and a transient increase in osteoblastic markers expression while pparγ and neuropilin-1 decreased. Hematopoietic supportive capacities of CFU-Fs were also modified: as compared to controls, irradiated CFU-Fs significantly increased the proliferation rate of hematopoietic precursors and accelerated the differentiation toward the granulocytic lineage. Our data provide the first evidence of the key role exerted by the balance between osteoblasts and adipocytes in spontaneous bone marrow regeneration. First, (pre)osteoblast differentiation from MSC stimulated hematopoietic precursor's proliferation and granulopoietic regeneration. Then, in a second time (pre)osteoblasts progressively disappeared in favour of adipocytic cells which down regulated the proliferation and granulocytic differentiation and then contributed to a return to pre-irradiation conditions