Enterococcal surface protein Esp is not essential for cell adhesion and intestinal colonization of Enterococcus faecium in mice

<p>Abstract</p> <p>Background</p> <p><it>Enterococcus faecium </it>has globally emerged as a cause of hospital-acquired infections with high colonization rates in hospitalized patients. The enterococcal surface protein Esp, identified as a potential virulence factor, is specifically linked to nosocomial clonal lineages that are genetically distinct from indigenous <it>E. faecium </it>strains. To investigate whether Esp facilitates bacterial adherence and intestinal colonization of <it>E. faecium</it>, we used human colorectal adenocarcinoma cells (Caco-2 cells) and an experimental colonization model in mice.</p> <p>Results</p> <p>No differences in adherence to Caco-2 cells were found between an Esp expressing strain of <it>E. faecium </it>(E1162) and its isogenic Esp-deficient mutant (E1162Δ<it>esp</it>). Mice, kept under ceftriaxone treatment, were inoculated orally with either E1162, E1162Δ<it>esp </it>or both strains simultaneously. Both E1162 and E1162Δ<it>esp </it>were able to colonize the murine intestines with high and comparable numbers. No differences were found in the contents of cecum and colon. Both E1162 and E1162Δ<it>esp </it>were able to translocate to the mesenteric lymph nodes.</p> <p>Conclusion</p> <p>These results suggest that Esp is not essential for Caco-2 cell adherence and intestinal colonization or translocation of <it>E. faecium </it>in mice.</p

Genomic Investigation of Two Acinetobacter baumannii Outbreaks in a Veterinary Intensive Care Unit in The Netherlands

Acinetobacter baumannii is a nosocomial pathogen that frequently causes healthcare-acquired infections. The global spread of multidrug-resistant (MDR) strains with its ability to survive in the environment for extended periods imposes a pressing public health threat. Two MDR A. baumannii outbreaks occurred in 2012 and 2014 in a companion animal intensive care unit (caICU) in the Netherlands. Whole-genome sequencing (WGS) was performed on dog clinical isolates (n = 6), environmental isolates (n = 5), and human reference strains (n = 3) to investigate if the isolates of the two outbreaks were related. All clinical isolates shared identical resistance phenotypes displaying multidrug resistance. Multi-locus Sequence Typing (MLST) revealed that all clinical isolates belonged to sequence type ST2. The core genome MLST (cgMLST) results confirmed that the isolates of the two outbreaks were not related. Comparative genome analysis showed that the outbreak isolates contained different gene contents, including mobile genetic elements associated with antimicrobial resistance genes (ARGs). The time-measured phylogenetic reconstruction revealed that the outbreak isolates diverged approximately 30 years before 2014. Our study shows the importance of WGS analyses combined with molecular clock investigations to reduce transmission of MDR A. baumannii infections in companion animal clinics

Toll-Like Receptor 2 Impairs Host Defense in Gram-Negative Sepsis Caused by Burkholderia pseudomallei (Melioidosis)

Willem Wiersinga and colleagues find up-regulation of multiple Toll-like receptors (TLRs) in peripheral blood cells of patients with melioidosis. However, only TLR2 had an effect on the immune response in a mouse model

Lipoteichoic acid-induced lung inflammation depends on TLR2 and the concerted action of TLR4 and the platelet-activating factor receptor

Lipoteichoic acid (LTA) is a major outer cell wall component of Gram-positive bacteria that has been implicated as an important factor in the inflammatory response following bacterial infection. In vitro data indicate roles for TLR2, platelet-activating factor receptor (PAFR), CD14, and LPS-binding protein (LBP) in cellular responsiveness to LTA, whereas the mechanisms contributing to LTA effects in vivo have never been investigated. Using mice deficient for LBP, CD14, TLR2, TLR4, or PAFR, we now examined the role of these molecules in pulmonary inflammation induced by highly purified LTA in vivo. Although pulmonary LBP increased dose-dependently following administration of LTA, the inflammatory response was unaltered in LBP-/- mice. TLR2 proved to be indispensable for the initiation of an inflammatory response, as polymorphonuclear cell influx, TNF-alpha, keratinocyte-derived chemokine, and MIP-2 release were abolished in TLR2-/- mice. Minor effects such as moderately decreased TNF-alpha and MIP-2 levels were observed in the absence of CD14, indicating a role for CD14 as a coreceptor. Quite surprisingly, the absence of TLR4 greatly diminished pulmonary inflammation and the same phenotype was observed in PAFR-/- animals. In contrast to all other mice studied, only TLR4-/- and PAFR-/- mice displayed significantly elevated IL-10 pulmonary concentrations. These data suggest that TLR2 is the single most important receptor signaling the presence of LTA within the lungs in vivo, whereas TLR4 and PAFR may influence lung inflammation induced by LTA either by sensing LTA directly or through recognition and signaling of endogenous mediators induced by the interaction between LTA and TLR

Intestinal Enterococcus faecium Colonization Improves Host Defense during Polymicrobial Peritonitis

Background. Vancomycin-resistant (VR) Enterococcus faecium is increasingly found to colonize and infect hospitalized patients. Enterococci are frequently isolated from polymicrobial infections originating from the intestines. The impact of VR E. faecium on these infections and vice versa is not clear. Methods. Mice were intestinally colonized with VR E. faecium during oral vancomycin treatment; control mice received oral vancomycin only. Fourteen days later, cecal ligation and puncture (CLP) was performed in all mice to induce polymicrobial peritonitis in the presence or absence of VR E. faecium colonization. Results. VR E. faecium colonization per se was not associated with systemic dissemination of VR E. faecium. CLP resulted in systemic VR E. faecium infection in all VR E. faecium-colonized mice, with high VR E. faecium loads in peritoneal lavage fluid, blood, liver, and lungs. Forty-eight hours after CLP, mice infected with VR E. faecium had significantly lower bacterial loads in all organs tested than mice not infected with VR E. faecium. Additionally, lower inflammatory parameters were measured in VR E. faecium-infected mice. CLP induced transient liver and kidney damage, with a faster recovery in VR E. faecium-colonized mice. Conclusions. VR E. faecium infection, originating from a natural source (the intestinal tract), does not worsen the outcome of CLP-induced polymicrobial peritonitis and sepsis but rather facilitates bacterial clearance and attenuates host inflammatory response

The Complement System Facilitates Clearance of Enterococcus faecium during Murine Peritonitis

Background. Infections with multidrug-resistant enterococci are a growing problem worldwide. Little is known about the host defense against enterococcal diseases. In vitro studies have demonstrated an important role played by complement proteins in neutrophil-mediated phagocytosis. In this study, we investigated the importance of complement in an in vivo model of Enterococcus faecium peritonitis. Methods. Peripheral neutrophils and peritoneal macrophages were incubated with E. faecium that had been preincubated with decomplemented or normal plasma, and phagocytosis and killing were examined. E. faecium peritonitis was induced in C57BL/6 mice rendered complement deficient by intraperitoneal injection with cobra venom factor (CVF) and in complement 3 (C3) knockout mice. The course of the infection was compared with that in saline control and wild-type mice, respectively, at several time points up to 48 h after infection. Results. Opsonization by complement enhanced phagocytosis by neutrophils and macrophages. CVF-treated and C3 knockout mice were severely hampered in clearing E. faecium from all organs and tissues under study (peritoneal fluid, blood, lungs, and liver). Higher peritoneal cytokine and chemokine levels were measured in decomplemented mice, whereas no differences in systemic or peritoneal cell kinetics were detected. Conclusion. Complement deficiency severely hampers the clearance of E. faecium peritonitis and subsequent systemic infectio