A simple phenotypic method for screening of MCR-1-mediated colistin resistance

Objectives: To evaluate a novel method, the colistin-MAC test, for phenotypic screening of acquired colistin resistance mediated by transferable mcr-1 resistance determinants, based on colistin MIC reduction in the presence of dipicolinic acid (DPA). Methods: The colistin-MAC test consists in a broth microdilution method, in which colistin MIC is tested in the absence or presence of DPA (900 \u3bcg/mL). Overall, 74 colistin-resistant strains of Enterobacteriaceae (65 Escherichia coli and nine other species), including 61 strains carrying mcr-1-like genes and 13 strains negative for mcr genes, were evaluated with the colistin-MAC test. The presence of mcr-1-like and mcr-2-like genes was assessed by real-time PCR and end-point PCR. For 20 strains, whole-genome sequencing data were also available. Results: A 658-fold reduction of colistin MIC in the presence of DPA was observed with 59 mcr-1-positive strains, including 53 E. coli of clinical origin, three E. coli transconjugants carrying MCR-1-encoding plasmids, one Enterobacter cloacae complex and two Citrobacter spp. Colistin MICs were unchanged, increased or at most reduced by twofold with the 13 mcr-negative colistin-resistant strains (nine E. coli and four Klebsiella pneumoniae), but also with two mcr-1-like-positive K. pneumoniae strains. Conclusions: The colistin-MAC test could be a simple phenotypic test for presumptive identification of mcr-1-positive strains among isolates of colistin-resistant E. coli, based on a 658-fold reduction of colistin MIC in the presence of DPA. Evaluation of the test with a larger number of strains, species and mcr-type resistance determinants would be of interest

Vaginal lactobacilli and vaginal dysbiosis-associated bacteria differently affect cervical epithelial and immune homeostasis and anti-viral defenses

Persistent infection with High Risk-Human Papilloma Viruses (HR-HPVs) is a primary cause of cervical cancer worldwide. Vaginal-dysbiosis-associated bacteria were correlated with the persistence of HR-HPVs infection and with increased cancer risk. We obtained strains of the most represented bacterial species in vaginal microbiota and evaluated their effects on the survival of cervical epithelial cells and immune homeostasis. The contribution of each species to supporting the antiviral response was also studied. Epithelial cell viability was affected by culture supernatants of most vaginal-dysbiosis bacteria, whereas Lactobacillus gasseri or Lactobacillus jensenii resulted in the best stimulus to induce interferon-γ (IFN-γ) production by human mononuclear cells from peripheral blood (PBMCs). Although vaginal-dysbiosis-associated bacteria induced the IFN-γ production, they were also optimal stimuli to interleukin-17 (IL-17) production. A positive correlation between IL-17 and IFN-γ secretion was observed in cultures of PBMCs with all vaginal-dysbiosis-associated bacteria suggesting that the adaptive immune response induced by these strains is not dominated by T(H)1 differentiation with reduced availability of IFN-γ, cytokine most effective in supporting virus clearance. Based on these results, we suggest that a vaginal microbiota dominated by lactobacilli, especially by L. gasseri or L. jensenii, may be able to assist immune cells with clearing HPV infection, bypasses the viral escape and restores immune homeostasis

The shadow position sensors (SPS) formation flying metrology subsystem for the ESA PROBA-3 mission: present status and future developments

PROBA-3 [1] [2] is a Mission of the European Space Agency (ESA) composed of two formation-flying satellites, planned for their joint launch by the end of 2018. Its main purposes have a dual nature: scientific and technological. In particular, it is designed to observe and study the inner part of the visible solar corona, thanks to a dedicated coronagraph called ASPIICS (Association of Spacecraft for Polarimetric and Imaging Investigation of the Corona of the Sun), and to demonstrate the in-orbit formation flying (FF) and attitude control capability of its two satellites. The Coronagraph payload on-board PROBA-3 consists of the following parts: the Coronagraph Instrument (CI) with the Shadow Position Sensor (SPS) on the Coronagraph Spacecraft (CSC), the Occulter Position Sensor (OPSE) [3] [4] and the External Occulting (EO) disk on the Occulter Spacecraft (OSC). The SPS subsystem [5] is one of the main metrological devices of the Mission, adopted to control and to maintain the relative (i.e. between the two satellites) and absolute (i.e. with respect to the Sun) FF attitude. It is composed of eight micro arrays of silicon photomultipliers (SiPMs) [6] that shall be able to measure, with the required sensitivity and dynamic range as asked by ESA, the penumbral light intensity on the Coronagraph entrance pupil. With the present paper we describe the testing activities on the SPS breadboard (BB) and Development Model (DM) as well as the present status and future developments of this PROBA-3 metrological subsystem

A chemical survey of exoplanets with ARIEL

Thousands of exoplanets have now been discovered with a huge range of masses, sizes and orbits: from rocky Earth-like planets to large gas giants grazing the surface of their host star. However, the essential nature of these exoplanets remains largely mysterious: there is no known, discernible pattern linking the presence, size, or orbital parameters of a planet to the nature of its parent star. We have little idea whether the chemistry of a planet is linked to its formation environment, or whether the type of host star drives the physics and chemistry of the planet’s birth, and evolution. ARIEL was conceived to observe a large number (~1000) of transiting planets for statistical understanding, including gas giants, Neptunes, super-Earths and Earth-size planets around a range of host star types using transit spectroscopy in the 1.25–7.8 μm spectral range and multiple narrow-band photometry in the optical. ARIEL will focus on warm and hot planets to take advantage of their well-mixed atmospheres which should show minimal condensation and sequestration of high-Z materials compared to their colder Solar System siblings. Said warm and hot atmospheres are expected to be more representative of the planetary bulk composition. Observations of these warm/hot exoplanets, and in particular of their elemental composition (especially C, O, N, S, Si), will allow the understanding of the early stages of planetary and atmospheric formation during the nebular phase and the following few million years. ARIEL will thus provide a representative picture of the chemical nature of the exoplanets and relate this directly to the type and chemical environment of the host star. ARIEL is designed as a dedicated survey mission for combined-light spectroscopy, capable of observing a large and well-defined planet sample within its 4-year mission lifetime. Transit, eclipse and phase-curve spectroscopy methods, whereby the signal from the star and planet are differentiated using knowledge of the planetary ephemerides, allow us to measure atmospheric signals from the planet at levels of 10–100 part per million (ppm) relative to the star and, given the bright nature of targets, also allows more sophisticated techniques, such as eclipse mapping, to give a deeper insight into the nature of the atmosphere. These types of observations require a stable payload and satellite platform with broad, instantaneous wavelength coverage to detect many molecular species, probe the thermal structure, identify clouds and monitor the stellar activity. The wavelength range proposed covers all the expected major atmospheric gases from e.g. H2O, CO2, CH4 NH3, HCN, H2S through to the more exotic metallic compounds, such as TiO, VO, and condensed species. Simulations of ARIEL performance in conducting exoplanet surveys have been performed – using conservative estimates of mission performance and a full model of all significant noise sources in the measurement – using a list of potential ARIEL targets that incorporates the latest available exoplanet statistics. The conclusion at the end of the Phase A study, is that ARIEL – in line with the stated mission objectives – will be able to observe about 1000 exoplanets depending on the details of the adopted survey strategy, thus confirming the feasibility of the main science objectives.Peer reviewedFinal Published versio

Hypervirulent Klebsiella pneumoniae Strains Modulate Human Dendritic Cell Functions and Affect TH1/TH17 Response

none13noHypervirulent Klebsiella pneumoniae (Hv‐Kp) strains have emerged as pathogens causing life‐threatening, invasive disease even in immunocompetent hosts. Systemic dissemination usually occurs following perturbations of the gut microbiota and is facilitated by Hv‐Kp resistance to phagocytosis and complement activity. Hv‐Kp are usually associated with K1 or K2 capsular types, produce several iron uptake systems (e.g., aerobactin and salmochelin) and are often but not invariably, capsular material hyper‐producers (hypermucoviscous phenotype: HMV). Whether Hv‐ Kp escape the immune response at mucosal site is unknown. In this work, we studied the effects of Hv‐Kp on human dendritic cells (DCs), central players of the IL‐23/IL‐17 and IL‐12/IFN‐γ axis at mucosal sites, essential for pathogen clearance. Four Hv‐Kp and HMV strains were selected and their activity on DC maturation and cytokine production was compared to that of non‐virulent Kp strains with classic or HMV phenotypes. While the maturation process was equally induced by all Kp strains, significant differences between virulent and non‐virulent strains were found in the expression of genes for cytokines involved in T‐cell activation and differentiation. The non‐virulent KP04C62 and the classic Kp, KPC157 induced high expression of TH1 (IL‐12p70 and TNFα) and TH17 cytokines (IL‐23, IL‐1β and IL‐6), while Hv‐Kp poorly activated these cytokine genes. Moreover, conditioned media from DCs cultured with non‐virulent Kp, either classical or hypercapsulated, induced the activation of IL‐17 and IFN‐γ genes in preactivated CD4+‐cells suggesting their TH17/TH1 differentiation. Conditioned media from Hv‐Kp poorly activated IL‐17 and IFN‐γ genes. In summary, our data indicate that Hv‐Kp interfere with DC functions and T‐cell differentiation and suggest that the escape from the IL‐23/IL‐17 and IL‐12/IFN‐γ axes may contribute to pathogen dissemination in immunocompetent hosts.noneNicolo S.; Mattiuz G.; Antonelli A.; Arena F.; Di Pilato V.; Giani T.; Baccani I.; Clemente A.M.; Castronovo G.; Tanturli M.; Cozzolino F.; Rossolini G.M.; Torcia M.G.Nicolo, S.; Mattiuz, G.; Antonelli, A.; Arena, F.; Di Pilato, V.; Giani, T.; Baccani, I.; Clemente, A. M.; Castronovo, G.; Tanturli, M.; Cozzolino, F.; Rossolini, G. M.; Torcia, M. G

Detection of oxazolidinone resistance genes and characterization of genetic environments in enterococci of Swine origin, Italy

One hundred forty-five florfenicol-resistant enterococci, isolated from swine fecal samples collected from 76 pig farms, were investigated for the presence of optrA, cfr, and poxtA genes by PCR. Thirty florfenicol-resistant Enterococcus isolates had at least one linezolid resistance gene. optrA was found to be the most widespread linezolid resistance gene (23/30), while cfr and poxtA were detected in 6/30 and 7/30 enterococcal isolates, respectively. WGS analysis also showed the presence of the cfr(D) gene in Enterococcus faecalis (n = 2 isolates) and in Enterococcus avium (n = 1 isolate). The linezolid resistance genes hybridized both on chromosome and plasmids ranging from ~25 to ~240 kb. Twelve isolates were able to transfer linezolid resistance genes to enterococci recipient. WGS analysis displayed a great variability of optrA genetic contexts identical or related to transposons (Tn6628 and Tn6674), plasmids (pE035 and pWo27-9), and chromosomal regions. cfr environments showed identities with Tn6644-like transposon and a region from p12-2300 plasmid; cfr(D) genetic contexts were related to the corresponding region of the plasmid 4 of Enterococcus faecium E8014; poxtA was always found on Tn6657. Circular forms were obtained only for optrA-and poxtA-carrying genetic contexts. Clonality analysis revealed the presence of E. faecalis (ST16, ST27, ST476, and ST585) and E. faecium (ST21) clones previously isolated from humans. These results demonstrate a dissemination of linezolid resistance genes in enterococci of swine origin in Central Italy and confirm the spread of linezolid resistance in animal settings

Clinical features and outcomes of bloodstream infections caused by New Delhi metallo-\u3b2-lactamase-producing enterobacterales during a regional outbreak

Limited data about New Delhi metallo-\u3b2-lactamase (NDM) bacteremia are available. Blood isolates from 40 patients with NDM bacteremia were studied for antibiotic susceptibility and whole-genomic sequencing. NDM bacteremia has high 30-day mortality. In most cases, aztreonam-avibactam is active in vitro. Ceftazidime-avibactam plus aztreonam may represent a feasible therapeutic option