Eco-Friendly vs. Traditional Cleaning in Healthcare Settings: Microbial Safety and Environmental Footprint Riccardo Fontana, Mattia Buratto, Anna Caproni, Chiara Nordi, Mariangela Pappadà, et al. Hygiene, 2025 Growing concern for environmental sustainability has resulted in the implementation of sanitization methods that respect ecological principles. This research evaluates a “green” sanitizing protocol that uses CAM (Minimum Environmental Criteria)-compliant products against a traditional protocol within two ASL Roma 1 facilities. The study performed a Life Cycle Assessment (LCA) following ISO 14040, ISO 14044, and ISO 14067 standards to measure greenhouse gases emissions. Microbiological sampling was conducted according to established protocols across three different risk zones utilizing contact plates and surface swabs. The Life Cycle Assessment showed that CO2 emissions reduced by 49.6% to 53.3% at different sites due to reduced energy use together with concentrated detergents and improved washing cycles. Microbiological testing revealed notable decreases in contamination rates across both cleaning systems yet demonstrated the “green” system achieved superior results specifically within high-risk zones. The “green” protocol matched traditional cleaning methods hygienically but delivered significant environmental advantages which positions it as a sustainable hospital cleaning solution.
Environmental Microbiological Sampling in Civil Settings: Comparative LCA Analysis of Green Cleaning Techniques vs. Traditional Methods in Accordance with New Italian CAM Guidelines Riccardo Fontana, Luciano Vogli, Mattia Buratto, Anna Caproni, Chiara Nordi, et al. Sustainability Switzerland, 2025 This research conducts a comparative life cycle assessment (LCA) to examine both the ecological footprint and microbiological performance of eco-friendly and traditional cleaning methods in non-clinical environments. Conducted in accordance with the updated Minimum Environmental Criteria (CAMs), the research follows the principles and framework established by UNI EN ISO 14040 and 14044. Additionally, the assessment of carbon footprint (kg CO2e) adheres to ISO 14067:2018, with guidance from Product Category Rules (PCR 2011:03 v3.0.1). Our findings demonstrate that the Green Protocol implemented by Dussmann Service reduces the carbon footprint of cleaning activities by 20.0% compared to the traditional protocol, equating to an annual reduction of 156 kg CO2 at the pilot site. Laboratory analyses confirm that the Green Protocol maintains hygiene standards equivalent to conventional methods, ensuring adequate microbiological quality while significantly lowering environmental impact. The study highlights the feasibility of integrating eco-friendly cleaning practices without compromising effectiveness. Future research should explore the scalability, cost-efficiency, and long-term benefits of this approach. This assessment provides a scientifically validated foundation for adopting sustainable cleaning methodologies in professional settings, supporting the transition towards environmentally responsible facility management.
Sustainable vs. Conventional Cleaning in Healthcare: Microbiological and Life Cycle Insights Riccardo Fontana, Luciano Vogli, Mattia Buratto, Anna Caproni, Chiara Nordi, et al. Sustainability Switzerland, 2025 This study investigates the environmental and microbiological effectiveness of two cleaning protocols—EVA (Ecological Visible Approach) and a traditional cleaning protocol—implemented in a healthcare setting. The cleaning procedures were conducted using specific equipment tailored to each protocol, with the EVA Protocol emphasizing sustainable practices. Microbiological evaluations assessed the effectiveness of each protocol in reducing microbial presence on various surfaces. The results demonstrated that the EVA Protocol not only met but often exceeded the microbiological standards achieved by the traditional protocol while significantly reducing the environmental footprint. The comparative Life Cycle Assessment (LCA) highlighted the EVA Protocol’s ability to lower CO2 emissions by 31.5% compared to the traditional protocol. This study underscores the potential benefits of adopting sustainable cleaning practices in healthcare settings, contributing to both environmental preservation and effective infection control.
Herpes Simplex Virus ICP27 Protein Inhibits AIM 2-Dependent Inflammasome Influencing Pro-Inflammatory Cytokines Release in Human Pigment Epithelial Cells (hTert-RPE 1) Anna Caproni, Chiara Nordi, Riccardo Fontana, Martina Facchini, Sara Melija, et al. International Journal of Molecular Sciences, 2024 Although Herpes simplex virus type 1 (HSV-1) has been deeply studied, significant gaps remain in the fundamental understanding of HSV-host interactions: our work focused on studying the Infected Cell Protein 27 (ICP27) as an inhibitor of the Absent-in-melanoma-2 (AIM 2) inflammasome pathway, leading to reduced pro-inflammatory cytokines that influence the activation of a protective innate immune response to infection. To assess the inhibition of the inflammasome by the ICP27, hTert-immortalized Retinal Pigment Epithelial cells (hTert-RPE 1) infected with HSV-1 wild type were compared to HSV-1 lacking functional ICP27 (HSV-1∆ICP27) infected cells. The activation of the inflammasome by HSV-1∆ICP27 was demonstrated by quantifying the gene and protein expression of the inflammasome constituents using real-time PCR and Western blot. The detection of the cleavage of the pro-caspase-1 into the active form was performed by using a bioluminescent assay, while the quantification of interleukins 1β (IL-1β) and 18 (IL-18)released in the supernatant was quantified using an ELISA assay. The data showed that the presence of the ICP27 expressed by HSV-1 induces, in contrast to HSV-1∆ICP27 vector, a significant downregulation of AIM 2 inflammasome constituent proteins and, consequently, the release of pro-inflammatory interleukins into the extracellular environment reducing an effective response in counteracting infection.
Evaluating Cleaning Services in Civil Environments: Microbiological and Life Cycle Analysis Comparing Conventional and Sustainable Methods Riccardo Fontana, Mattia Buratto, Anna Caproni, Chiara Nordi, Mariangela Pappadà, et al. Sustainability Switzerland, 2024 In response to the growing global concern for environmental sustainability, a Life Cycle Assessment (LCA) study was conducted to evaluate the environmental benefits of the “Formula Servizi” GREEN protocol compared to a conventional cleaning protocol, as mandated by the updated Criteria for Environmental Sustainability (CAM—Criteri Ambientali Minimi) for cleaning services. The CAM, effective on 19 June 2021, requires companies to demonstrate the environmental advantages of their cleaning protocols over traditional methods. This study aligns with the new CAM guidelines and employs UNI EN ISO 14040–14044 technical standards for a comprehensive comparative analysis. The study highlights the significance of maintaining hygiene to ensure safety in various contexts, emphasizing the importance of environmental sampling and monitoring to prevent contamination and infection transmission. Despite the complexity and expenses associated with microbiological monitoring, this research affirms its crucial role in validating cleaning procedures, particularly in healthcare facilities, food service areas, and industrial settings. The findings reveal that both the “Traditional” and “GREEN” cleaning protocols demonstrate satisfactory effectiveness in controlling microbiological contamination according to established guidelines. Moreover, the LCA results indicate that the “GREEN” protocol, while exhibiting higher water consumption and wastewater treatment, showcases a strategic use of more sustainable cleaning and laundry detergents. Despite the increased water usage in certain phases, the significantly lower environmental impact per unit of weight demonstrates the potential for optimizing both environmental sustainability and operational efficiency in future Life Cycle Sustainability Assessment (LCSA) endeavors. The comparative LCA further reveals that the “GREEN” protocol enables an annual avoidance of 260 g of CO2-e emissions per square meter of cleaned surface. The most significant reduction in absolute terms is associated with the use of eco-labeled detergents in the laundry system, resulting in the avoidance of 654.1 kg of CO2-e per year of service (−77% compared to traditional laundry detergents).
Suppressing gain-of-function proteins via CRISPR/Cas9 system in SCA1 cells Mariangela Pappadà, Ottavia Bonuccelli, Mattia Buratto, Riccardo Fontana, Mariaconcetta Sicurella, et al. Scientific Reports, 2022 SCAs are autosomal dominant neurodegenerative disorders caused by a gain-of-function protein with toxic activities, containing an expanded polyQ tract in the coding region. There are no treatments available to delay the onset, stop or slow down the progression of these pathologies. In this work we focus our attention on SCA1 which is one of the most common genotypes circulating in Italy. Here, we develop a CRISPR/Cas9-based approach to reduce both forms of the ATXN1 protein, normal and mutated with expanded polyQ. We started with the screening of 10 different sgRNAs able to target Exon 8 of the ATXN1 gene. The two most promising sgRNAs were validated in fibroblasts isolated from SCA1 patients, following the identification of the best transfection method for this type of cell. Our silencing approach significantly downregulated the expression of ataxin1, due to large deletions and the introduction of small changes in the ATXN1 gene, evidenced by NGS analysis, without major effects on cell viability. Furthermore, very few significant guide RNA-dependent off-target effects were observed. These preliminary results not only allowed us to identify the best transfection method for SCA1 fibroblasts, but strongly support CRISPR/Cas9 as a promising approach for the treatment of expanded polyQ diseases. Further investigations will be needed to verify the efficacy of our silencing system in SCA1 neurons and animal models.
Control of Erwinia amylovora Growth by Moringa oleifera Leaf Extracts: In Vitro and in Planta Effects Riccardo Fontana, Giovanna Macchi, Anna Caproni, Mariaconcetta Sicurella, Mattia Buratto, et al. Plants, 2022 Erwinia amylovora (EA) is a phytopathogenic bacterium, the causative agent of bacterial fire blight, a disease that affects Rosaceaes. In order to replace antibiotics and copper, the antimicrobial activity of three extracts of Moringa oleifera Lam., methanolic (MeOH-MOE), hydroalcoholic (HA-MOE) and hydroalcoholic with maltodextrins (HAMD-MOE), was tested on eleven strains of EA isolated from apple trees by the Emilia-Romagna Phytosanitary Department. MIC and MBC have been evaluated; biofilm formation, swarming motility and amylovoran production were performed with the crystalviolet, soft-agar assay and the amylovoran method. All extracts demonstrated bacteriostatic activity at a concentration of 1 mg/mL, resulting in a 80% reduction in biofilm formation. HAMD-MOE, MeOH-MOE and HA-MOE caused an inhibition of motility of 60%, 65% and 30% after 6 days and a decrease in amylovoran synthesis of 84%, 63% and 93%, respectively. In planta results showed how the compounds were able to inhibit EA virulence on apple trees, mainly if they were applied as a preventive treatment, although the treatment showed a significant reduction in fire blight symptoms progression. The antibacterial activity of the extracts is mainly due to the high concentration of polyphenolic compounds detected in the extracts that was able to alter the permeability of bacterial membrane, resulting in slowing the synthesis of ATP and consequently of all ATP-dependent functions, such as motility and less selectivity towards harmful compounds, which can, thus, enter the cytoplasm and inhibit enzymes involved in replication and quorum sensing. The efficacy, eco-compatibility and low cost make such extracts a potential tool for the control of bacterial fire blight.
Effects of moringa oleifera leaf extracts on xanthomonas campestris pv. Campestris Riccardo Fontana, Anna Caproni, Raissa Buzzi, Mariaconcetta Sicurella, Mattia Buratto, et al. Microorganisms, 2021 Xanthomonas campestris pv. campestris (Xcc) is a Gram-negative bacterium belonging to the Xanthomonodaceae family, causing black rot in crucifers. To control this pathogen, the study investigated the effect of different leaves extracts of Moringa oleifera Lam., a tropical plant, well known for its food properties and with countless applications in many different fields, from nutraceutical (hypoglycemic) to the cosmetic (sunscreen) properties. Nevertheless, several studies pointed to its antibacterial action against both Gram-negative and Gram-positive bacteria. Many bioactive compounds, including flavonoids, phenolic acids, alkaloids, isothiocyanates, tannins and saponins, contained in these extracts, are responsible for its countless activities. The analyses carried out in this study show that the methanolic, hydroalcoholic and hydroalcoholic maltodextrin extracts have both bacteriostatic and bactericidal effects at concentrations of 0.5, 0.5 and 0.1 mg/mL respectively. In particular, the study shows how all extracts can alter membrane permeability, to adversely affect swarming motility, and to alter biofilm formation in Xcc. The in planta experiments showed a reduction of the necrosis area in the infected radishes, although the ability of the extracts to be absorbed by root systems is yet to be understood, in order to reach the target point.
UPF1 silenced cellular model systems for screening of read-through agents active on β 0 39 thalassemia point mutation Francesca Salvatori, Mariangela Pappadà, Giulia Breveglieri, Elisabetta D’Aversa, Alessia Finotti, et al. BMC Biotechnology, 2018 BackgroundNonsense mutations promote premature translational termination, introducing stop codons within the coding region of mRNAs and causing inherited diseases, including thalassemia. For instance, in β039 thalassemia the CAG (glutamine) codon is mutated to the UAG stop codon, leading to premature translation termination and to mRNA destabilization through the well described NMD (nonsense-mediated mRNA decay). In order to develop an approach facilitating translation and, therefore, protection from NMD, ribosomal read-through molecules, such as aminoglycoside antibiotics, have been tested on mRNAs carrying premature stop codons. These findings have introduced new hopes for the development of a pharmacological approach to the β039 thalassemia therapy. While several strategies, designed to enhance translational read-through, have been reported to inhibit NMD efficiency concomitantly, experimental tools for systematic analysis of mammalian NMD inhibition by translational read-through are lacking.ResultsWe developed a human cellular model of the β039 thalassemia mutation with UPF-1 suppressed and showing a partial NMD suppression.ConclusionsThis novel cellular model could be used for the screening of molecules exhibiting preferential read-through activity allowing a great rescue of the mutated transcripts.
