Candida spp. are an important opportunistic pathogen that can represent a possible cause of severe infections, especially in immunocompromised individuals. The clinical impact of Candida spp. depends, in part, on the ability to form biofilms, communities of nestled cells into the extracellular matrix. In this study, we compared the biofilm formation ability of 83 strains of Candida spp. isolated from blood cultures and other materials, such as respiratory samples, urine, and exudate, and their sensitivity to fluconazole (FLZ). Strains were divided into tertiles to establish cut-offs to classify isolates as low, moderate, or high biofilm producers (0.839) and biofilms with low, moderate, or high metabolic activity (0.183). A non-linear relationship between biofilm production and metabolic activity was found in C. glabrata and C. tropicalis. In addition, the increase in minimum biofilm eradication concentrations (MBEC₅₀) compared to the Minor Inhibitory Concentration (PMIC) of the planktonic form in Candida spp. confirms the role of biofilm in the induction of resistance to FLZ.

Recent Monkeypox virus (MPXV) outbreaks in non-endemic regions have highlighted the need for genomic surveillance to support epidemiological investigations and monitor viral evolution. In this paper we present the results of genomic characterization and analyses of mechanisms of human adaptation, including APOBEC-style mutations, performed on 11 MPXV isolates, collected from May to September 2022, from Emilia-Romagna (Italy). Phylogenetic analysis confirmed all strains belonged to Clade IIb. Viruses from male patients were classified within lineage B (sub-lineages B.1, B.1.3, B.1.12), while a strain from a female patient was assigned to lineage A (A.2.3), with epidemiological links to Ghana. This represents the fourth detection of an A.2.3 strain of African origin outside the continent. Disruptions were identified in two genes: OPG176 (similar to VACV-Cop A46R), as reported in lineage A, and OPG023 (similar to VACV-Cop D7L), resulting in protein truncation that may suggest a possible pattern of human adaptation. These findings further enhance our understanding of MPXV Clade IIb diversity through characterization of a rarer variant giving insights from a less explored epidemiological pathway.

Developing low-cost vaccine production strategies is crucial to achieving global health equity and mitigating the spread and impact of disease outbreaks. High hydrostatic pressure (HHP) technology is a widely used technology employed in the food industry for long-term preservation. This project aims at validating HHP as a cost-effective method for the production of highly immunogenic thermal stable whole-virus SARS-CoV-2 vaccines. Structural studies on HHP-inactivated viruses demonstrated pressure-dependent effects, with higher pressures (500–600 MPa) destabilizing viral morphology. Immunogenicity assessments, in animal models, revealed that 500 MPa treatment elicited the most robust humoral and cellular immune responses, outperforming heat inactivation. Additionally, HHP-inactivated viral preparation retained thermostability for 30 days at 4 °C, reducing cold-chain dependencies and enabling vaccine distribution also in low-resource settings. With its rapid, cost-effective, and scalable production process, HHP presents a transformative, equitable solution for global vaccine development, particularly for emerging pathogens.

Staphylococcus argenteus, identified in 2006, represents a challenging case of bacterial taxonomic identification because of its high similarity to Staphylococcus aureus. In this context, neither mass spectrometry (MS) nor 16S gene analysis cannot precisely reveal the difference between the two species. In our study, the sensitivity to antibiotics of S. argenteus isolated from blood culture was tested, and the investigation of the bacterial genome was performed by Multi-Locus Sequence Typing (MLST) and Whole-Genome Next-Generation Sequencing (WG-NGS). The pathogen was identified as ST2250 and presented perfectly matched resistance genes, namely aph(3′)-III, mgrA, and sepA, whereas the virulence gene detected was scn. Two plasmids were found: the pSAS plasmid, belonging to the family of Inc18, and plasmid pN315, belonging to the Rep3 group. The epidemiological distribution and the spread of S. argenteus infection are scarcely documented, particularly when associated with sepsis. Therefore, a correct taxonomy identification, antibiogram, and resistance gene analysis may help in acquiring knowledge about this bacterium and implement its detection and treatment.

Fever of Unknown Origin (FUO) is broadly defined as a fever with an unidentified cause despite a minimum set of diagnostic investigations. The variability of FUO etiologies across geographic areas, age groups, and decades makes diagnosis challenging and complicates the adoption of a standardized diagnostic approach. Global warming and changing interactions between humans, animals, and the environment are contributing to the emergence and re-emergence of zoonotic infections. Emerging Vector-Borne Disease (VBD) pathogens circulate in Northern Italy, but their prevalence and impact on febrile illnesses remain poorly understood. This study investigated FUO epidemiology, diagnosis, and treatment in three hospitals in Emilia-Romagna (northeastern Italy). The medical records from 652 patients who were discharged with the International Classification of Diseases, Ninth Revision (ICD-9) codes "780.6 Fever of unknown origin" and "087.9 Relapsing fever, unspecified" between January 2017 and December 2023 were analysed. Among patients discharged with FUO between 2017 and 2023, the mean age was 58 years, and 45 % older than 65 years. Comorbidities were present in 75 %, and 26 % had active cancer. A diagnostic hypothesis was present in 32 % of cases. Diagnostic tests were mainly laboratory-based; 5.9 % had confirmed infections. Antibiotics were used in 62 % of patients mostly penicillin/beta-lactamase inhibitors. FUO discharges peaked in summer. Active hospital-based surveillance are crucial to deepen our current understanding on FUO epidemiology and possible contribution of VBD pathogens while refining the use of antibiotics in the clinical practice.

Toscana virus (TOSV), a Phlebovirus transmitted by sandflies, is a leading cause of aseptic meningitis in the Mediterranean region. Despite its clinical significance, underreporting and limited availability of complete genomic data hinder a thorough understanding of its genetic diversity and evolution. This study presents a novel amplicon-based whole-genome sequencing (WGS) method using Illumina library preparation kits and proprietary software to optimize workflows and enhance bioinformatic analyses. Primers targeting TOSV lineage A genomes were designed with PrimalScheme to generate 400 bp amplicons, incorporating degenerate bases to improve coverage. Library preparation utilized Illumina Microbial Amplicon Prep (iMAP) kits, followed by de novo assembly using BaseSpace DRAGEN Targeted Microbial software. The method’s sensitivity was tested on viral propagates at various RNA concentrations (10⁴ to 10 copies/μL), demonstrating robust performance at concentrations above 10² copies/μL. Validation on high-titre viral propagates (n = 7), low-titre clinical samples (n = 15), and phlebotomine pools (n = 5) confirmed its reproducibility and ability to comprehensively cover coding regions. Cerebrospinal fluid samples yielded the most consistent results compared to urine and sandfly pools. This innovative WGS approach represents a significant advancement in TOSV genomic surveillance, enabling large-scale studies of its genetic diversity and evolutionary dynamics, which are critical for improving diagnostics and public health strategies.