Multicenter evaluation of the vitek MS matrix-assisted laser desorption ionization-time of flight mass spectrometry system for identification of gram-positive aerobic bacteria

Matrix-assisted laser desorption ionization–time of flight mass spectrometry (MALDI-TOF) is gaining momentum as a tool for bacterial identification in the clinical microbiology laboratory. Compared with conventional methods, this technology can more readily and conveniently identify a wide range of organisms. Here, we report the findings from a multicenter study to evaluate the Vitek MS v2.0 system (bioMérieux, Inc.) for the identification of aerobic Gram-positive bacteria. A total of 1,146 unique isolates, representing 13 genera and 42 species, were analyzed, and results were compared to those obtained by nucleic acid sequence-based identification as the reference method. For 1,063 of 1,146 isolates (92.8%), the Vitek MS provided a single identification that was accurate to the species level. For an additional 31 isolates (2.7%), multiple possible identifications were provided, all correct at the genus level. Mixed-genus or single-choice incorrect identifications were provided for 18 isolates (1.6%). Although no identification was obtained for 33 isolates (2.9%), there was no specific bacterial species for which the Vitek MS consistently failed to provide identification. In a subset of 463 isolates representing commonly encountered important pathogens, 95% were accurately identified to the species level and there were no misidentifications. Also, in all but one instance, the Vitek MS correctly differentiated Streptococcus pneumoniae from other viridans group streptococci. The findings demonstrate that the Vitek MS system is highly accurate for the identification of Gram-positive aerobic bacteria in the clinical laboratory setting

Bacterial contamination of inanimate surfaces and equipment in the intensive care unit

Intensive care unit (ICU)-acquired infections are a challenging health problem worldwide, especially when caused by multidrug-resistant (MDR) pathogens. In ICUs, inanimate surfaces and equipment (e.g., bedrails, stethoscopes, medical charts, ultrasound machine) may be contaminated by bacteria, including MDR isolates. Cross-transmission of microorganisms from inanimate surfaces may have a significant role for ICU-acquired colonization and infections. Contamination may result from healthcare workers' hands or by direct patient shedding of bacteria which are able to survive up to several months on dry surfaces. A higher environmental contamination has been reported around infected patients than around patients who are only colonized and, in this last group, a correlation has been observed between frequency of environmental contamination and culture-positive body sites. Healthcare workers not only contaminate their hands after direct patient contact but also after touching inanimate surfaces and equipment in the patient zone (the patient and his/her immediate surroundings). Inadequate hand hygiene before and after entering a patient zone may result in cross-transmission of pathogens and patient colonization or infection. A number of equipment items and commonly used objects in ICU carry bacteria which, in most cases, show the same antibiotic susceptibility profiles of those isolated from patients. The aim of this review is to provide an updated evidence about contamination of inanimate surfaces and equipment in ICU in light of the concept of patient zone and the possible implications for bacterial pathogen cross-transmission to critically ill patients

Heat Resistance Mediated by a New Plasmid Encoded Clp ATPase, ClpK, as a Possible Novel Mechanism for Nosocomial Persistence of Klebsiella pneumoniae

Klebsiella pneumoniae is an important opportunistic pathogen and a frequent cause of nosocomial infections. We have characterized a K. pneumoniae strain responsible for a series of critical infections in an intensive care unit over a two-year period. The strain was found to be remarkably thermotolerant providing a conceivable explanation of its persistence in the hospital environment. This marked phenotype is mediated by a novel type of Clp ATPase, designated ClpK. The clpK gene is encoded by a conjugative plasmid and we find that the clpK gene alone renders an otherwise sensitive E. coli strain resistant to lethal heat shock. Furthermore, one third of a collection of nosocomial K. pneumoniae isolates carry clpK and exhibit a heat resistant phenotype. The discovery of ClpK as a plasmid encoded factor and its profound impact on thermal stress survival sheds new light on the biological relevance of Clp ATPases in acquired environmental fitness and highlights the challenges of mobile genetic elements in fighting nosocomial infections

Decrypting magnetic fabrics (AMS, AARM, AIRM) through the analysis of mineral shape fabrics and distribution anisotropy

The fieldwork was supported by the DIPS project (grant no. 240467) and the MIMES project (grant no. 244155) funded by the Norwegian Research Council awarded to O.G. O.P.'s position was funded from Y-TEC.Anisotropy of magnetic susceptibility (AMS) and anisotropy of magnetic remanence (AARM and AIRM) are efficient and versatile techniques to indirectly determine rock fabrics. Yet, deciphering the source of a magnetic fabric remains a crucial and challenging step, notably in the presence of ferrimagnetic phases. Here we use X-ray micro-computed tomography to directly compare mineral shape-preferred orientation and spatial distribution fabrics to AMS, AARM and AIRM fabrics from five hypabyssal trachyandesite samples. Magnetite grains in the trachyandesite are euhedral with a mean aspect ratio of 1.44 (0.24 s.d., long/short axis), and > 50% of the magnetite grains occur in clusters, and they are therefore prone to interact magnetically. Amphibole grains are prolate with magnetite in breakdown rims. We identified three components of the petrofabric that influence the AMS of the analyzed samples: the magnetite and the amphibole shape fabrics and the magnetite spatial distribution. Depending on their relative strength, orientation and shape, these three components interfere either constructively or destructively to produce the AMS fabric. If the three components are coaxial, the result is a relatively strongly anisotropic AMS fabric (P’ = 1.079). If shape fabrics and/or magnetite distribution are non-coaxial, the resulting AMS is weakly anisotropic (P’ = 1.012). This study thus reports quantitative petrofabric data that show the effect of magnetite distribution anisotropy on magnetic fabrics in igneous rocks, which has so far only been predicted by experimental and theoretical models. Our results have first-order implications for the interpretation of petrofabrics using magnetic methods.Publisher PDFPeer reviewe

Listeria pathogenesis and molecular virulence determinants

The gram-positive bacterium Listeria monocytogenes is the causative agent of listeriosis, a highly fatal opportunistic foodborne infection. Pregnant women, neonates, the elderly, and debilitated or immunocompromised patients in general are predominantly affected, although the disease can also develop in normal individuals. Clinical manifestations of invasive listeriosis are usually severe and include abortion, sepsis, and meningoencephalitis. Listeriosis can also manifest as a febrile gastroenteritis syndrome. In addition to humans, L. monocytogenes affects many vertebrate species, including birds. Listeria ivanovii, a second pathogenic species of the genus, is specific for ruminants. Our current view of the pathophysiology of listeriosis derives largely from studies with the mouse infection model. Pathogenic listeriae enter the host primarily through the intestine. The liver is thought to be their first target organ after intestinal translocation. In the liver, listeriae actively multiply until the infection is controlled by a cell-mediated immune response. This initial, subclinical step of listeriosis is thought to be common due to the frequent presence of pathogenic L. monocytogenes in food. In normal indivuals, the continual exposure to listerial antigens probably contributes to the maintenance of anti-Listeria memory T cells. However, in debilitated and immunocompromised patients, the unrestricted proliferation of listeriae in the liver may result in prolonged low-level bacteremia, leading to invasion of the preferred secondary target organs (the brain and the gravid uterus) and to overt clinical disease. L. monocytogenes and L. ivanovii are facultative intracellular parasites able to survive in macrophages and to invade a variety of normally nonphagocytic cells, such as epithelial cells, hepatocytes, and endothelial cells. In all these cell types, pathogenic listeriae go through an intracellular life cycle involving early escape from the phagocytic vacuole, rapid intracytoplasmic multiplication, bacterially induced actin-based motility, and direct spread to neighboring cells, in which they reinitiate the cycle. In this way, listeriae disseminate in host tissues sheltered from the humoral arm of the immune system. Over the last 15 years, a number of virulence factors involved in key steps of this intracellular life cycle have been identified. This review describes in detail the molecular determinants of Listeria virulence and their mechanism of action and summarizes the current knowledge on the pathophysiology of listeriosis and the cell biology and host cell responses to Listeria infection. This article provides an updated perspective of the development of our understanding of Listeria pathogenesis from the first molecular genetic analyses of virulence mechanisms reported in 1985 until the start of the genomic era of Listeria research

Time to intestinal clearance of carbapenemase-producing Enterobacterales in hospital patients: a longitudinal retrospective observational cohort study

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