Sabrina Semeraro

Background Waterlogged archaeological deposits can preserve skin- and fur-based artefacts that are otherwise rarely recovered, yet their taxonomic attribution and biochemical integrity are often uncertain and difficult to evaluate by morphology alone. Here, a remarkably preserved fragment of hair-bearing pelt recovered during a 2014 rescue excavation in the medieval urban core of Tønsberg (Norway) and radiocarbon-dated to ca. 1200 CE was investigated to clarify its biological origin and preservation state through an integrated analytical workflow. Results Microscopy documented the overall structural integrity of the hair and skin tissues, while FT-IR spectroscopy supported the assessment of collagen preservation and informed post-excavation handling. Bottom-up proteomics (nanoLC–MS/MS, Orbitrap) narrowed species attribution to caprines, with peptide evidence favouring Ovis aries over Capra hircus and identification of sheep-specific marker peptides in the hair fraction. Independent ancient DNA analysis targeting a short mitochondrial COI fragment (mini-COI) and high-throughput sequencing confirmed 100% identity with Ovis aries, with Capra sequences showing substantially lower similarity. Significance By converging protein- and DNA-based evidence, this study provides a robust species identification for a rare medieval pelt find and demonstrates the value of proteogenomic integration for perishable cultural heritage materials, supporting more reliable interpretation, documentation, and conservation of fragile organic artefacts recovered from wet contexts.

Surface-enhanced Raman scattering (SERS) spectroscopy has grown in popularity as a bioaerosol monitoring method due to its high sensitivity and specificity, as well as its ability to be performed in complex biological mixtures using portable and relatively inexpensive devices. However, due to a lack of standardised methodologies, SERS sensing of bioaerosols remains difficult. Full-length peer-reviewed journal articles related to the application of SERS spectroscopy to examine bioaerosols were systematically searched in PubMed, Scopus, and Web of Science databases using the PRISMA guidelines. A total of 13 studies met the inclusion criteria for our systematic literature search. A critical evaluation of the experimental aspects involved in the collection of bioaerosols for SERS analysis is presented, as well as the elective applicability and weaknesses of various experimental setups, helping to provide a solid foundation for real-time bioaerosol characterisation using SERS spectroscopy.

This study presents a comprehensive multimodal analytical study of an Egyptian ritual Bes-vase, of the 2nd century BCE employing cutting-edge proteomics, metabolomics, genetics techniques, and synchrotron radiation-based Fourier Transformed Infrared microSpectroscopy (SR µ-FTIR) to characterize organic residues of its content. We successfully identified the presence of various functional, bioactive, psychotropic, and medicinal substances, shedding light on the diverse components of a liquid concoction used for ritual practices in Ptolemaic Egypt. Using LC-MS/MS with a new methodological approach, we identified key proteins and metabolites, enabling the identification of botanical sources, confirmed by genetic sequences. Our analyses revealed traces of Peganum harmala, Nimphaea nouchali var. caerulea, and a plant of the Cleome genus, all of which are traditionally proven to have psychotropic and medicinal properties. Additionally, the identification of human fluids suggests their direct involvement in these rituals. Furthermore, metabolomics and SR µ-FTIR analyses also revealed the presence of fermented fruit-based liquid and other ingredients such as honey or royal jelly. The identification of specific chemical compounds, such as alkaloids and flavonoids, provides insight into the psychoactive and therapeutic uses of these in ancient ritual practices. This multidisciplinary study highlights the complexity of ancient cultures and their interactions with psychoactive, medicinal, and bioactive substances. These findings contribute to our understanding of ancient belief systems, cultural practices, and the utilization of natural resources, ultimately enhancing our knowledge of past societies and their connection to the natural world.

In the wake of the COVID-19 pandemic, the surveillance and safety measures of indoor Cultural Heritage sites have become a paramount concern due to the unique challenges posed by their enclosed environments and high visitor volumes. This communication explores the integration of Artificial Intelligence (AI) in enhancing epidemiological surveillance and health safety protocols in these culturally significant spaces. AI technologies, including machine learning algorithms and Internet of Things (IoT) sensors, have shown promising potential in monitoring air quality, detecting pathogens, and managing crowd dynamics to mitigate the spread of infectious diseases. We review various applications of AI that have been employed to address both direct health risks and indirect impacts such as visitor experience and preservation practices. Additionally, this paper discusses the challenges and limitations of AI deployment, such as ethical considerations, privacy issues, and financial constraints. By harnessing AI, Cultural Heritage sites can not only improve their resilience against future pandemics but also ensure the safety and well-being of visitors and staff, thus preserving these treasured sites for future generations. This exploration into AI’s role in post-COVID surveillance at Cultural Heritage sites opens new frontiers in combining technology with traditional conservation and public health efforts, providing a blueprint for enhanced safety and operational efficiency in response to global health challenges.

The escalation of the COVID-19 outbreak has significantly increased research into the transmission of airborne infectious diseases in indoor settings, underscoring the urgent necessity for affordable and efficient methods of air disinfection. The aim of the present work is the development of a complete framework based on designed experiments for exploring and optimizing the bioaerosol removal and inactivation efficiency of a novel air disinfection device. This device combines the aerodynamic effect of a three-dimensional vortex structure with UV-C radiation provided by commercially available UV-C light-emitting diodes (UV-C LEDs). The system was designed and tested to locally maintain a high radiation intensity that is suitable for bioaerosol disinfection. A controlled experimental laboratory model of bioaerosol aerosolization was set up by using an impinger medical vibrating nebulizer, a cylindrical chamber for bioaerosol travel, and an SKC BioSampler for collecting microorganisms capable of replicating. A nonpathogenic strain of E. coli (BL21-DE3) was used as a model of airborne bacteria. The inactivation efficiency was assessed based on the enumeration of the colonies originating from viable E. coli. Interactions between analytical factors and their optimal levels were investigated by using sequential D-optimal designs adapted to domain constraints and previous computational simulations of the aerodynamic performance of the device. Five experimental variables (the concentration of aerosolized bacteria; the size of aerosol particles; the volumetric airflow; the power of the LEDs; and two configurations of the device) were considered as factors in the optimization process. Response surfaces allowed for the identification of the ideal working conditions to maximize the efficiency of the device, an essential requirement for the device’s future exploitation in real-world settings.

Studies on bioaerosol bacterial biodiversity have relevance in both ecological and health contexts, and molecular methods, such as 16S rRNA gene-based barcoded sequencing, provide efficient tools for the analysis of airborne bacterial communities. Standardized methods for sampling and analysis of bioaerosol DNA are lacking, thus hampering the comparison of results from studies implementing different devices and procedures. Three samplers that use gelatin filtration, swirling aerosol collection, and condensation growth tubes for collecting bioaerosol at an aeration tank of a wastewater treatment plant in Trieste (Italy) were used to determine the bacterial biodiversity. Wastewater samples were collected directly from the untreated sewage to obtain a true representation of the microbiological community present in the plant. Different samplers and collection media provide an indication of the different grades of biodiversity, with condensation growth tubes and DNA/RNA shield^TM capturing the richer bacterial genera. Overall, in terms of relative abundance, the air samples have a lower number of bacterial genera (64 OTUs) than the wastewater ones (75 OTUs). Using the metabarcoding approach to aerosol samples, we provide the first preliminary step toward the understanding of a significant diversity between different air sampling systems, enabling the scientific community to orient research towards the most informative sampling strategy.

Background: Standardized methods for testing Viral Filtration Efficiency (VFE) of tissues and devices are lacking and few studies are available on aerosolizing, sampling and assessing infectivity of SARS-CoV-2 in controlled laboratory settings. NanoAg-coated endonasal filters appear a promising aid for lowering viable virus inhalation in both adult and younger populations (e.g., adolescents). Objective: to provide an adequate method for testing SARS-CoV-2 bioaerosol VFE of bio-gel Ag nanoparticles endonasal filters, by a model system, assessing residual infectivity as cytopathic effect and viral proliferation on in vitro cell cultures. Methods: A SARS-CoV-2 aerosol transmission chamber fed by a BLAM aerosol generator produces challenges (from very high viral loads (10⁵ PFU/mL) to lower ones) for endonasal filters positioned in a Y shape sampling port connected to a Biosampler. An aerosol generator, chamber and sampler are contained in a class II cabinet in a BSL3 facility. Residual infectivity is assessed from aliquots of liquid collecting bioaerosol, sampled without and with endonasal filters. Cytopathic effect as plaque formation and viral proliferation assessed by qRT-PCR on Vero E6 cells are determined up to 7 days post inoculum. Results: Each experimental setting is replicated three times and basic statistics are calculated. Efficiency of aerosolization is determined as difference between viral load in the nebulizer and in the Biosampler at the first day of experiment. Efficiency of virus filtration is calculated as RNA viral load ratio in collected bioaerosol with and without endonasal filters at the day of the experiment. Presence of infectious virus is assessed by plaque forming unit assay and RNA viral load variations. Conclusions: A procedure and apparatus for assessing SARS-CoV-2 VFE for endonasal filters is proposed. The apparatus can be implemented for more sophisticated studies on contaminated aerosols.

In recent years, antimicrobial peptides (AMPs) have enjoyed a renaissance, as the world is currently facing an emergency in terms of severe infections that evade antibiotics’ treatment. This is due to the increasing emergence and spread of resistance mechanisms. Covalent conjugation with polymers is an interesting strategy to modulate the pharmacokinetic profile of AMPs and enhance their biocompatibility profile. It can also be an effective approach to develop active coatings for medical implants and devices, and to avoid biofilm formation on their surface. In this concise review, we focus on the last 5 years’ progress in this area, pertaining in particular to AMPs that contain d-amino acids, as well as their role, and the advantages that may arise from their introduction into AMPs.

The airborne route of transmission of SARS-CoV-2 was confirmed by the World Health Organization in April 2021. There is an urge to establish standardized protocols for assessing the concentration of SARS-CoV-2 RNA in air samples to support risk assessment, especially in indoor environments. Debates on the airborne transmission route of SARS-CoV-2 have been complicated because, among the studies testing the presence of the virus in the air, the percentage of positive samples has often been very low. In the present study, we report preliminary results on a study for the evaluation of parameters that can influence SARS-CoV-2 RNA recovery from quartz fiber filters spotted either by standard single-stranded SARS-CoV-2 RNA or by inactivated SARS-CoV-2 virions. The analytes were spiked on filters and underwent an active or passive sampling; then, they were preserved at −80 °C for different numbers of days (0 to 54) before extraction and analysis. We found a mean recovery of 2.43%, except for the sample not preserved (0 days) that showed a recovery of 13.51%. We found a relationship between the number of days and the recovery percentage. The results presented show a possible issue that relates to the quartz matrix and SARS-CoV-2 RNA recovery. The results are in accordance with the already published studies that described similar methods for SARS-CoV-2 RNA field sampling and that reported non-detectable concentrations of RNA. These outcomes could be false negatives due to sample preservation conditions. Thus, until further investigation, we suggest, as possible alternatives, to keep the filters: (i) in a sealed container for preservation at 4 °C; and (ii) in a viral transport medium for preservation at a temperature below 0 °C.

Severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2) is mainly transmitted through respiratory droplets, aerosols, or direct contact with fomites from an infected subject. It has been reported that SARS-CoV-2 is stable and viable in aerosol up to 16 h in controlled laboratory conditions. However, the aerosolization conditions varied a lot between the studies. In this work, an experimental laboratory model of SARS-CoV-2 aerosolization was established, employing an impinger nebulizer, a cylindrical chamber for aerosol travel, and a SKC biosampler for the collection of particles. The efficiency of the system was assessed based on the molecular determination of the viral load in the nebulizer after the aerosolization and in the aerosol collected at the end of the travel. Moreover, the residual infectivity was tested in vitro on the Vero E6 cell line, through the observation of the cytopathic effect (CPE), and the quantification of the viral load in the supernatants at 7 days post inoculation (dpi). A high RNA viral load was found in the SKC biosampler after aerosolization, indicating that it was possible to transport a high virus titer through the 30-cm chamber with all the dilutions (initial 10⁵, 10⁴, 10³ plaque forming unit—PFU/mL). At the 7 dpi, an increment of the RNA viral load was determined for the dilutions 10⁵ and 10⁴ PFU/mL tested, while only the initial 10⁵ PFU/mL resulted in visible CPE. Our findings allowed us to achieve the resilience of SARS-CoV-2 in aerosol form, at a concentration comparable to those reported for clinical samples. This mode of transmission should be considered for the mitigation and preventive measures to counteract SARS-CoV-2 spreading.