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Legionella pneumophila is a freshwater opportunistic pathogen and the leading cause of severe pneumonia known as Legionnaires' disease. It can be found in all water systems and survives in biofilms, free-living amoebae, and a wide variety of facilities, such as air conditioning and showers in hospitals, hotels and spas. The reference cultural method allows for the isolation and identification in many days, and in addition, it does not detect viable but rather non-culturable bacteria, increasing the risk of infection. In this context, a new LAMP-based (loop-mediated isothermal amplification) kit was developed, allowing for the rapid, sensitive, and labor-saving detection of L. pneumophila . The kit, " Legionella pneumophila Glow", was validated according to ISO/TS 12869:2012, testing sensitivity, inclusivity and exclusivity, and kit robustness. Sensitivity showed that the " Legionella pneumophila Glow" kit can detect up to 28 plasmid copies/µL. Robustness tests showed consistent results, with both contamination levels and the matrices used giving reproducible results. Furthermore, real samples were evaluated to compare the performance of the two methods. The LAMP kit " Legionella pneumophila Glow" proved a useful option for the rapid, efficient, and labor-saving screening of different typologies of water samples, offering significant advantages over the traditional method, as it is characterized by a high sensitivity, ease of use for laboratory testing, and a large reduction in analysis time, making it an asset to official controls.

Introduction : Surveillance of Candida species isolates from blood cultures (BCs) in Europe is considered fragmented, unable to allow the definition of targets of antifungal stewardship recommendations especially during the SARS-CoV-2 pandemic. Methods : We performed a multicentric retrospective study including all consecutive BC Candida isolates from six Southern European tertiary hospitals (1st January 2020 to 31st December 2021). Etiology, antifungal susceptibility patterns, and clinical setting were analyzed and compared. Results : C. albicans was the dominant species (45.1%), while C. auris was undetected. Candida species positive BC events increased significantly in COVID-19 ICUs in 2021 but decreased in other ICUs. Resistance to azole increased significantly and remained very high in C. albicans (fluconazole from 0.7% to 4.5%, p = 0.03) and C. parapsilosis complex (fluconazole up to 24.5% and voriconazole up to 8.9%), respectively. Resistance to caspofungin was remarkable in C. tropicalis (10%) and C. krusei (20%), while resistance to at least one echinocandin increased in 2021, especially in C. parapsilosis complex (from 0.8% to 5.1%, p = 0.05). Although no significant differences were observed over the study period, fluconazole and echinocandin resistance increased in COVID-19 ICUs by up to 14% and 5.8%, respectively, but remained undetected in non-intensive COVID-19 wards. Conclusions : Antifungal stewardship activities aimed at monitoring resistance to echinocandin in C. tropicalis and C. krusei , and against the spread of fluconazole resistant C. parapsilosis complex isolates are highly desirable. In COVID-19 patients, antifungal resistance was mostly present when the illness had a critical course.

C. difficile enterocolitis (CDAC) is the most common hospital infection, burdened by an increased incidence of coagulation-related complications such as deep vein thrombosis (DVT) and disseminated intravascular coagulation (DIC) as well as a significant sepsis-related mortality. In this review, we analyzed the available data concerning the correlation between coagulation complications related to C. difficile infection (CDI) and inflammasome activation, in particular the pyrin-dependent one. The little but solid available preclinical and clinical evidence shows that inflammasome activation increases the risk of venous thromboembolism (VTE). As proof of this, it has been observed that in vitro inhibition of the molecules (e.g., tissue factor) mainly involved in coagulation activation could block the process. In vivo studies show that it could be possible to reduce the incidence of complications associated with C. difficile infection (CDI) and mortality due to a state of hypercoagulability. A personalized therapeutic approach to reduce the inflammatory activity and prevent thromboembolic complications could be preliminarily defined to reduce mortality.

Legionella is a ubiquitous bacterium that lives in freshwater environments and colonizes human-made water systems. Legionella pneumophila is the most virulent species, and risk factors for Legionnaires' disease include increasing age, smoking, chronic diseases, and immunodeficiency. For this reason, it is very important to assess and monitor hospital water systems in order to prevent legionellosis. We have monitored a large hospital in Palermo for four years. To determine the presence of microorganisms, according to national guidelines, we used the culture method, which is considered the gold standard for Legionella detection. Sampling was divided into five macro-areas, and a total of 251 samples were collected during the period of investigation, 49% of which were Legionella spp.-positive and 51% were Legionella spp.-negative. Positive samples with L. pneumophila . sgr 2-15 were most frequent in the Underground (55.6%, p = 0.0184), Medicine (42.9%, p = 0.0184) and Other (63.2%, p = 0.002) areas; while positive samples for L. pneumophila sgr 1 were less frequent in the Underground (0.0%, p = 0.0184) and Surgery areas (4.5%, p = 0.033), and for Legionella anisa , were less frequent in the Medicine (4.1%, p = 0.021), Oncohematology (0.0%, p = 0.0282), and Other (0.0%, p = 0.016) areas. Finally, no significant differences were observed among the areas for each isolate considered. The surveillance carried out in these years demonstrates the importance of monitoring, which allows us to analyze the conditions of hospital facilities and, therefore, prevent Legionella spp. infections.