Strong epidemiological evidence suggests that onchocerciasis may be associated with epilepsy—hence the name onchocerciasis-associated epilepsy (OAE). However, the pathogenesis of OAE still needs to be elucidated, as recent studies have failed to detect Onchocerca volvulus in the central nervous system of persons with OAE. Therefore, it was suggested that a potentially neurotropic virus transmitted by blackflies could play a role in triggering OAE. To investigate this hypothesis, adult blackflies were collected in an onchocerciasis-endemic area with a high OAE prevalence in the Ntui Health District, Cameroon. A viral particle-based shotgun sequencing approach was used to detect viral sequences in 55 pools of 10 blackflies. A very high abundance of viral reads was detected across multiple (novel) viral families, including viral families associated with human disease. Although no genomes closely related to known neurotropic viruses were found in the blackfly virome, the plethora of novel viruses representing novel species, genera and even families warrant further exploration for their potential to infect vertebrates. These results could serve as a first step for studying the viruses associated with the haematophagous blackfly, which also could be present in their nematode host O. volvulus. Exploring the diversity of viruses in blackflies should be included in the active surveillance of zoonotic diseases.

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.

Transboundary animal diseases (TADs), including zoonoses, can be introduced in non‐endemic areas through animal trade and uncontrolled movements during times of conflict and migration. Sheep and goat pox (SGPX) and peste des petits ruminants (PPRs) were, respectively, reported in Europe in 2018 and 2010. EFSA emphasised the risk of spread into Europe through informal trade and unmonitored migration routes that may involve the informal transport of small ruminants. Using a One Health conceptual framework, which considers the interconnectedness of human, animal, and environmental health, this study analysed the epidemic trends of the two selected diseases during 2014–2025, with particular focus on the role of human migrations in influencing their spread along major migratory routes in the Balkan Peninsula. The conceptualisation of animals in migration settings was investigated through the scrutiny of policies and semi‐structured interviews to experts and people who travelled the route. The presence of small ruminants, companion and pest animals was confirmed in formal and informal camps. The results underscore the relevance of animals in the migration contexts of the Balkan routes, as well as the risks associated with their presence in settlements in the absence of an adequate management. Our findings seem to suggest that human migration along the Balkan routes is not directly responsible for the spread of TADs, as there is no evidence of significant animal movements accompanying migrants. However, our results show a possible role of informal trade networks following human migration paths along the Balkan route, while a possible role of informal trade networks, following human migration paths along the Balkan route, deserves to be further investigated.

Acute infectious gastroenteritis (AIG) is caused by diverse bacterial and viral enteric pathogens with overlapping clinical presentations, limiting conventional pathogen-directed workflows. This retrospective before–after study evaluated the diagnostic, microbiological, operational, and economic impact of multiplex molecular syndromic testing for AIG within a regional hub-and-spoke laboratory network in Italy. Two 19-month periods were compared: a pre-implementation period based on conventional diagnostics and a post-implementation period using multiplex syndromic panels as first-line tests. Diagnostic investigations totalled 25,574 before and 24,509 after implementation. Overall test positivity increased from 5.43% to 11.59% (p < 0.001), with significant increases for bacterial and viral targets. Pathogen detection events increased from 1388 to 2839, including broader detection of Campylobacter spp., Shigella spp./enteroinvasive Escherichia coli (EIEC), Yersinia enterocolitica, Aeromonas spp., Astrovirus, Sapovirus, and Norovirus genogroups. Mean turnaround time decreased from 63 to 47 h from sample collection and from 56 to 41 h from laboratory check-in. Although direct diagnostic costs increased moderately, total laboratory costs decreased when personnel costs were included, and cost per diagnostic detection declined from €233.80 to €120.86. Multiplex syndromic testing improved microbiological detection, diagnostic efficiency, turnaround time, and laboratory organisation in routine AIG diagnosis.

Accurate hepatitis B virus (HBV) DNA quantification is essential for assessing viral replication and monitoring antiviral therapy. This retrospective method-comparison study evaluated the Bosphore HBV Quantification Kit against the Aptima HBV Quant Assay using 130 residual anonymized EDTA-plasma specimens, comprising 110 comparator-positive and 20 comparator-negative samples. Categorical agreement was assessed using positive, negative, and overall percent agreement (PPA, NPA, and OPA), while quantitative agreement was evaluated using a predefined ±0.5 log₁₀ IU/mL criterion, Bland–Altman analysis, Passing–Bablok regression, and Lin’s concordance correlation coefficient (CCC). PPA was 100% (110/110; 95% confidence interval [CI], 96.70–100.00%), NPA was 95.0% (19/20; 95% CI, 75.13–99.87%), and OPA was 99.23% (129/130; 95% CI, 95.79–99.98%). One confirmed qualitative discordance was observed. Overall, 109/110 paired numerical results (99.1%; 95% CI, 95.04–99.98%) were within ±0.5 log₁₀ IU/mL. Bland–Altman analysis showed a mean bias of +0.124 log₁₀ IU/mL, Passing–Bablok regression yielded Y = 0.107 + 1.019X, and Lin’s CCC was 0.991. The Bosphore assay demonstrated high categorical and quantitative agreement with the comparator across the HBV DNA concentration range evaluated, supporting its potential suitability for quantitative HBV DNA testing within the analytical workflow assessed in this study.