Wastewater-Based Epidemiology for Viral Surveillance from an Endemic Perspective: Evidence and Challenges.

Wastewater-based epidemiology (WBE) is currently used to monitor not only the spread of the viral SARS-CoV-2 pandemic but also that of other viruses in endemic conditions, particularly in the absence of syndromic surveillance. The continuous monitoring of sewage requires high expenditure and significant time investments, highlighting the need for standardized methods and structured monitoring strategies. In this context, we conducted weekly wastewater monitoring in northwestern Tuscany (Italy) and targeted human adenovirus (HAdV), norovirus genogroup II (NoVggII), enterovirus (EV), and SARS-CoV-2. Samples were collected at the entrances of treatment plants and concentrated using PEG/NaCl precipitation, and viral nucleic acids were extracted and detected through real-time reverse transcription qPCR. NoVggII was the most identified target (84.4%), followed by HAdV, SARS-CoV-2, and EV. Only HAdV and EV exhibited seasonal peaks in spring and summer. Compared with data that were previously collected in the same study area (from February 2021 to September 2021), the results for SARS-CoV-2 revealed a shift from an epidemic to an endemic pattern, at least in the region under investigation, which was likely due to viral mutations that led to the spreading of new variants with increased resistance to summer environmental conditions. In conclusion, using standardized methods and an efficient monitoring strategy, WBE proves valuable for viral surveillance in pandemic and epidemic scenarios, enabling the identification of temporal-local distribution patterns that are useful for making informed public health decisions.

How to Evaluate If Microorganisms Isolated From Sterile Drug Production Environments Monitoring Are Undesirable.

This study addresses the identification of undesirable microorganisms (MOs) recovered during the environmental monitoring in manufacture of sterile medicinal products. We developed a methodology evaluation based on a decision tree; then, such approach was applied to hypothetical scenarios of uncommon MOs isolation in sterile drugs production settings. The scenarios were formulated on the basis of our field experience, in terms of both MOs selection and types of sampling site. The MOs were chosen in order to include emerging pathogens and MOs responsible for drug recall, and several sampling sites were considered for their detection (air, surfaces, and personnel). The classification of the unusual MOs revealed that most of them were undesirable, because they represented the loss of environmental control or a potential impact on the quality of the product. In some cases, the uncommon MOs were not considered as undesirable. Therefore, our results demonstrated the importance of a methodology, also in terms of recovery rate of unusual MOs and of the threshold probability for the unacceptability (e.g., 1% or 5%). The proposed methodology allowed an easy and documented evaluation for the undesirable MOs isolated from the environment of the analyzed settings for sterile drugs production.

Critical Needs for Integrated Surveillance: Wastewater-Based and Clinical Epidemiology in Evolving Scenarios with Lessons Learned from SARS-CoV-2.

During the COVID-19 pandemic, wastewater-based epidemiology (WBE) and clinical surveillance have been used as tools for analyzing the circulation of SARS-CoV-2 in the community, but both approaches can be strongly influenced by some sources of variability. From the challenging perspective of integrating environmental and clinical data, we performed a correlation analysis between SARS-CoV-2 concentrations in raw sewage and incident COVID-19 cases in areas served by medium-size wastewater treatment plants (WWTPs) from 2021 to 2023. To this aim, both datasets were adjusted for several sources of variability: WBE data were adjusted for factors including the analytical protocol, sewage flow, and population size, while clinical data adjustments considered the demographic composition of the served population. Then, we addressed the impact on the correlation of differences among sewerage networks and variations in the frequency and type of swab tests due to changes in political and regulatory scenarios. Wastewater and clinical data were significantly correlated when restrictive containment measures and limited movements were in effect (ρ = 0.50) and when COVID-19 cases were confirmed exclusively through molecular testing (ρ = 0.49). Moreover, a positive (although weak) correlation arose for WWTPs located in densely populated areas (ρ = 0.37) and with shorter sewerage lengths (ρ = 0.28). This study provides methodological approaches for interpreting WBE and clinical surveillance data, which could also be useful for other infections. Data adjustments and evaluation of possible sources of bias need to be carefully considered from the perspective of integrated environmental and clinical surveillance of infections.

Optimized numerical solutions of SIRDVW multiage model controlling SARS-CoV-2 vaccine roll out: An application to the Italian scenario.

In the context of SARS-CoV-2 pandemic, mathematical modelling has played a fundamental role for making forecasts, simulating scenarios and evaluating the impact of preventive political, social and pharmaceutical measures. Optimal control theory represents a useful mathematical tool to plan the vaccination campaign aimed at eradicating the pandemic as fast as possible. The aim of this work is to explore the optimal prioritisation order for planning vaccination campaigns able to achieve specific goals, as the reduction of the amount of infected, deceased and hospitalized in a given time frame, among age classes. For this purpose, we introduce an age stratified SIR-like epidemic compartmental model settled in an abstract framework for modelling two-doses vaccination campaigns and conceived with the description of COVID19 disease. Compared to other recent works, our model incorporates all stages of the COVID-19 disease, including death or recovery, without accounting for additional specific compartments that would increase computational complexity and that are not relevant for our purposes. Moreover, we introduce an optimal control framework where the model is the state problem while the vaccine doses administered are the control variables. An extensive campaign of numerical tests, featured in the Italian scenario and calibrated on available data from Dipartimento di Protezione Civile Italiana, proves that the presented framework can be a valuable tool to support the planning of vaccination campaigns. Indeed, in each considered scenario, our optimization framework guarantees noticeable improvements in terms of reducing deceased, infected or hospitalized individuals with respect to the baseline vaccination policy.

Organic Pollutants Associated with Plastic Debris in Marine Environment: A Systematic Review of Analytical Methods, Occurrence, and Characteristics.

Plastic pollution has become one of the most serious environmental problems, and microplastics (MPs, particles < 5 mm size) may behave as a vehicle of organic pollutants, causing detrimental effects to the environment. Studies on MP-sorbed organic pollutants lack methodological standardization, resulting in a low comparability and replicability. In this work, we reviewed 40 field studies of MP-sorbed organic contaminants using PRISMA guidelines for acquiring information on sampling and analytical protocols. The papers were also scored for their reliability on the basis of 7 criteria, from 0 (minimum) to 21 (maximum). Our results showed a great heterogeneity of the methods used for the sample collection, MPs extraction, and instruments for chemicals' identification. Measures for cross-contamination control during MPs analysis were strictly applied only in 13% of the studies, indicating a need for quality control in MPs-related research. The most frequently detected MP-sorbed chemicals were polychlorinated biphenyls (PCBs), polycyclic aromatic hydrocarbons (PAHs), and organochlorine pesticides (OCPs). Most of the studies showed a good reliability (>75% of the total score), with 32 papers scoring 16 or higher. On the basis of the collected information, a standardizable protocol for the detection of MPs and MP-sorbed chemicals has been suggested for improving the reliability of MPs monitoring studies.

In Vitro Potential Virucidal Effect Evaluation of Xibornol on Human Adenovirus Type 5, Human Rhinovirus Type 13, Human Coronavirus 229E, Human Parainfluenza Virus Type 1, and Human Respiratory Syncytial Virus.

The availability of virucidal compounds to reduce the impact of respiratory viruses is a relevant topic for public health, especially during the recent coronavirus disease (COVID-19) pandemic. Antimicrobial properties of Xibornol are known since the 1970s, but its activity on viruses is currently little explored. In this study, Xibornol activity at a fixed concentration of 0.03 mg/100 ml has been evaluated on five respiratory viruses (Human Adenovirus 5, Human Rhinovirus type 13, Human Coronavirus 229E, Human Parainfluenza Virus type 1, and Human Respiratory Syncytial Virus) through in vitro experiments based on adapted European standard UNI EN 14476-20019. The experiments were carried out under two different environmental conditions, one with the addition of fetal bovine serum to simulate an in vivo condition (dirty condition) and the other without the addition of any organic substances (clean condition). The viral abatement of Xibornol (expressed as Log10 reduction - LR) was statistically significant under both clean and dirty environmental conditions. Namely, in clean condition, LR ranged from 2.67 to 3.84, while in the dirty one the abatement was slightly lower (from 1.75 to 3.03). Parainfluenza Virus and Human Adenovirus were most resistant compared to the other viruses. The obtained data confirmed Xibornol activity and its use as topic substance for viral inactivation to prevent upper respiratory tract disease.

Calibration of Methods for SARS-CoV-2 Environmental Surveillance: A Case Study from Northwest Tuscany.

The current pandemic has provided an opportunity to test wastewater-based epidemiology (WBE) as a complementary method to SARS-CoV-2 monitoring in the community. However, WBE infection estimates can be affected by uncertainty factors, such as heterogeneity in analytical procedure, wastewater volume, and population size. In this paper, raw sewage SARS-CoV-2 samples were collected from four wastewater treatment plants (WWTPs) in Tuscany (Northwest Italy) between February and December 2021. During the surveillance period, viral concentration was based on polyethylene glycol (PEG), but its precipitation method was modified from biphasic separation to centrifugation. Therefore, in parallel, the recovery efficiency of each method was evaluated at lab-scale, using two spiking viruses (human coronavirus 229E and mengovirus vMC0). SARS-CoV-2 genome was found in 80 (46.5%) of the 172 examined samples. Lab-scale experiments revealed that PEG precipitation using centrifugation had the best recovery efficiency (up to 30%). Viral SARS-CoV-2 load obtained from sewage data, adjusted by analytical method and normalized by population of each WWTP, showed a good association with the clinical data in the study area. This study highlights that environmental surveillance data need to be carefully analyzed before their use in the WBE, also considering the sensibility of the analytical methods.

Virus contamination and infectivity in beach environment: Focus on sand and stranded material.

To assess the exposure of beachgoers to viruses, a study on seawater, sand, and beach-stranded material was carried out, searching for human viruses, fecal indicator organisms, and total fungi. Moreover, for the first time, the genome persistence and infectivity of two model viruses was studied in laboratory-spiked sand and seawater samples during a one-week experiment. Viral genome was detected in 13.6 % of the environmental samples, but it was not infectious (Human Adenovirus - HAdV, and enterovirus). Norovirus and SARS-CoV-2 were not detected. The most contaminated samples were from sand and close to riverine discharges. In lab-scale experiments, the infectivity of HAdV5 decreased by ~1.5-Log10 in a week, the one of Human Coronavirus-229E disappeared in <3 h in sand. The genome of both viruses persisted throughout the experiment. Our results confirm viral contamination of the beach and suggest HAdV as an index pathogen for beach monitoring and quantitative risk assessment.

Adding functions to marine infrastructure: Pollutant accumulation, physiological and microbiome changes in sponges attached to floating pontoons inside marinas.

The rate of introduction of man-made habitats in coastal environments is growing at an unprecedented pace, as a consequence of the expansion of urban areas. Floating installations, due to their unique hydrodynamic features, are able to provide great opportunities for enhancing water detoxification through the use of sessile, filtering organisms. We assessed whether the application of sponges to floating pontoons could function as a tool for biomonitoring organic and inorganic pollutants and for improving water quality inside a moderately contaminated marina in the NW Mediterranean. Fragments of two common Mediterranean sponges (Petrosia (Petrosia) ficiformis and Ircinia oros) were fixed to either suspended natural fibre nets beneath a floating pontoon or to metal frames deployed on the sea bottom. We assessed the accumulation of organic and inorganic contaminants in sponge fragments and, in order to provide an insight into their health status, we examined changes in their metabolic and oxidative stress responses and associated microbiomes. Fragments of both sponge species filtered out pollutants from seawater on both support types, but generally showed a better physiological and metabolic status when fixed to nets underneath the pontoon than to bottom frames. P. (P) ficiformis maintained a more efficient metabolism and exhibited a lower physiological stress levels and higher stability of the associated microbiome in comparison with I. oros. Our study suggests that the application of sponges to floating pontoon represents a promising nature-based solution to improve the ecological value of urban environments.

Exploring the Online Health Information-Seeking Behavior in a Sample of Italian Women: The "SEI Donna" Study.

There is much discussion about the skills of people in understanding and managing online health information. The Italian survey "SEI Donna" aimed to investigate perceptions and use of the web in women regarding health issues considering their health literacy (HL) and healthcare skills. We used an online questionnaire to explore different aspects of online health-related information-seeking behavior. The study participants (n = 7027) were categorized into healthcare workers (HW), healthcare students (HS), and non-healthcare women (non-HW). Half the sample (52%) searched online for a second opinion after the medical examination without statistical difference among HW, HS, and non-HW. Women in the age range of 26−40 years (OR = 1.28, p < 0.001), having chronic illness (OR = 1.48; p < 0.001), and being moderately (OR = 1.58; p < 0.001) or not satisfied (OR = 2.04; p < 0.001) with healthcare professionals were more likely to use the Internet to seek medical insight. Overall, 34% of women had a functional HL, the same being higher in HW (64%) and in HS (43%) than the rest of the women (18%) (p < 0.0001). The suboptimal HL suggests the need to improve HL in the general population to be skilled in surfing the web and, at the same time, to reorganize health training to improve the HL of healthcare professionals, also enriching their communication skills.

Impact of storms and proximity to entry points on marine litter and wrack accumulation along Mediterranean beaches: Management implications.

Beach litter can affect public health and economic activities worldwide forcing local authorities to expensive beach cleaning. Understanding the key mechanisms affecting the accumulation of this waste on beaches, such as sea state and proximity to entry points, is critical to plan effective management strategies. In this one-year study, we estimated the impact of storm events and waterways runoff on litter abundance and local economy using as a model a managed, peri-urban beach facing a north-western sector of the Mediterranean Sea. We also investigated the relationship between litter composition/density and beach proximity to major/closest harbors/rivers at regional scale by combining our data with those on litter density available in literature. Autumn/winter storms caused larger litter depositions than spring/summer ones in the peri-urban beach. No preferential accumulation occurred near to waterway mouths. Litter mainly consisted of plastic, and its composition in terms of micro-categories varied over seasons. In total, 367,070 items were deposited along 4.7 km of beach over one year, and the cost for the removal of this waste amounted to approximately 27,600 euros per km/year. At regional scale, beach litter density was positively correlated to the proximity of major harbors while its composition was related to the proximity to both major harbors and rivers. Results indicate that autumn/winter storms are important drivers of marine litter deposition. They also suggest that beaches in front of the convergence zone of littoral currents and close to major harbors can be particularly subjected to this kind of pollutant. To increase their effectiveness, litter mitigation/cleaning activities should be planned based on predictions of major storm events and performed at spatial scales encompassing at least coastal regional sectors.

Quantitative Microbial Risk Assessment Applied to Legionella Contamination on Long-Distance Public Transport.

The quantitative microbial risk assessment (QMRA) framework is used for assessing health risk coming from pathogens in the environment. In this paper, we used QMRA to evaluate the infection risk of L. pneumophila attributable to sink usage in a toilet cabin on Italian long-distance public transportation (LDT). LDT has water distribution systems with risk points for Legionella proliferation, as well as premise plumbing for drinking water, but they are not considered for risk assessment. Monitoring data revealed that approximately 55% of water samples (217/398) were positive for L. pneumophila, and the most frequently isolated was L. pneumophila sg1 (64%, 139/217); therefore, such data were fitted to the best probability distribution function to be used as a stochastic variable in the QMRA model. Then, a sink-specific aerosolization ratio was applied to calculate the inhaled dose, also considering inhalation rate and exposure time, which were used as stochastic parameters based on literature data. At L. pneumophila sg1 concentration ≤100 CFU/L, health risk was approximately 1 infection per 1 million exposures, with an increase of up to 5 infections per 10,000 exposures when the concentrations were ≥10,000 CFU/L. Our QMRA results showed a low Legionella infection risk from faucets on LDT; however, it deserves consideration since LDT can be used by people highly susceptible for the development of a severe form of the disease, owing to their immunological status or other predisposing factors. Further investigations could also evaluate Legionella-laden aerosols from toilet flushing.

Human adenovirus in municipal solid waste leachate and quantitative risk assessment of gastrointestinal illness to waste collectors.

Leachate is a variable effluent from waste management systems generated during waste collection and on landfills. Twenty-two leachate samples from waste collection trucks and a landfill were collected from March to December 2019 in the municipality of Rio de Janeiro (Brazil) and were analyzed for Human Adenovirus (HAdV), bacterial indicators and physico-chemical parameters. For viral analysis, samples were concentrated by ultracentrifugation and processed for molecular analysis using QIAamp Fast DNA Stool mini kit® for DNA extraction followed by nested-PCR and qPCR/PMA-qPCR TaqMan® system. HAdV was detected by nested-PCR in 100% (9/9) and 83.33% (12/13) of the truck and landfill leachate samples, respectively. Viral concentrations ranged from 8.31 × 101 to 6.68 × 107 genomic copies per 100 ml by qPCR and PMA-qPCR. HAdV species A, B, C, and F were characterized using nucleotide sequencing. HAdV were isolated in A549 culture cells in 100% (9/9) and 46.2% (6/13) from truck and landfill leachate samples, respectively. Regardless of the detection methods, HAdV concentration was predicted by the quantity of total suspended solids. A quantitative microbial risk assessment was performed to measure the probability of gastrointestinal (GI) illness attributable to inadvertent oral ingestion of truck leachate, revealing the higher probability of disease for the direct splashing into the oral cavity (58%) than for the gloved hand-to-mouth (33%). In a scenario where waste collectors do not wear gloves as protective personal equipment, the risk increases to 67%. This is the first study revealing infectious HAdV in solid waste leachate and indicates a potential health risk for waste collectors.

Objectionable microorganisms in pharmaceutical production: Validation of a decision tree.

The release of quality, safe, and effective non-sterile drugs needs to exclude the presence of objectionable microorganisms, which include microorganisms potentially involved in product degradation, or considered as poor hygiene indicator during manufacturing, or causing adverse effect on patient's health. In this paper, a method allowing objective and verifiable evaluations has been investigated through the development of a suitable decision tree with a template for data collection. The decision tree has been used to establish which microorganisms were objectionables, using several hypothetical scenarios in which 24 different biological agents, both harmless microorganisms and opportunistic pathogens, were combined with 9 different products, representing each type of administration route for non-sterile drugs. The results showed that the use of aforementioned approach makes the microorganisms evaluation easy and verifiable and highlighted that even the microbes initially considered harmless could be objectionable.

Pro-Environmental Behaviors: Determinants and Obstacles among Italian University Students.

The awareness of citizens concerning the health risks caused by environmental pollution is growing, but studies on determinants of pro-environmental behaviors have rarely examined health-related aspects. In this study, we investigated these determinants using data from a large survey among Italian university students (15 Universities: 4778 filled questionnaires). Besides the health-related aspects, represented by environmental health risk perception and functional health literacy, we considered social and demographic characteristics (gender, area of residence, sources of information, trust in institutional and non-institutional subjects, and students' capacity of positive actions, indicated as internal locus of control). The attitudes towards pro-environmental behaviors were positive for more than 70% of students and positively related with health risk perception, internal locus of control, and health literacy. The correspondence between the positive attitudes towards pro-environmental behaviors and the real adoption of such behaviors was approximately 20% for most behaviors, except for the separate collection of waste (60%). Such a discrepancy can be attributable to external obstacles (i.e., lack of time, costs, lack of support). The health-related aspects were linked to the pro-environmental attitudes, but to a lesser extent to pro-environmental behaviors, owing to the complexity of their determinants. However, they should be taken in account in planning education interventions.

Agri-Food Extracts Effectiveness in Improving Antibacterial and Antiviral Properties of Face Masks: A Proof-of-Concept Study.

The European dependencies for raw materials supply from foreign countries have been unquestionably shown by COVID-19 outbreak and have become particular evident from the slow response to the need for high quality personal protective equipment (PPEs). Among all medical devices, surgical face masks have earned themselves a primary role for the containment of the epidemic. In this context, our work aims at improving the barrier effect of surgical mask by depositing on their external surface a mixture of bioactive compounds, mainly polyphenols, extracted from agronomical sources. The main objective is the integration of the biorefining of agri-food solid wastes with the potential virucidal properties of the polyphenolic extracts for the treatment of PPEs.

Predicting needlestick and sharps injuries in nursing students: Development of the SNNIP scale.

Aim: To develop an instrument to investigate knowledge and predictive factors of needlestick and sharps injuries (NSIs) in nursing students during clinical placements. Design: Instrument development and cross-sectional study for psychometric testing. Methods: A self-administered instrument including demographic data, injury epidemiology and predictive factors of NSIs was developed between October 2018-January 2019. Content validity was assessed by a panel of experts. The instrument's factor structure and discriminant validity were explored using principal components analysis. The STROBE guidelines were followed. Results: Evidence of content validity was found (S-CVI 0.75; I-CVI 0.50-1.00). A three-factor structure was shown by exploratory factor analysis. Of the 238 participants, 39% had been injured at least once, of which 67.3% in the second year. Higher perceptions of "personal exposure" (4.06, SD 3.78) were reported by third-year students. Higher scores for "perceived benefits" of preventive behaviours (13.6, SD 1.46) were reported by second-year students.

Quantitative Microbial Risk Assessment as support for bathing waters profiling.

Profiling bathing waters supported by Quantitative Microbial Risk Assessment (QMRA) is key to the WHO's recommendations for the 2020/2021 revision of the European Bathing Water Directive. We developed an area-specific QMRA model on four pathogens, using fecal indicator concentrations (E. coli, enterococci) for calculating pathogen loads. The predominance of illness was found to be attributable to Human Adenovirus, followed by Salmonella, Vibrio, and Norovirus. Overall, the cumulative illness risk showed a median of around 1 case/10000 exposures. The risk estimates were strongly influenced by the indicators that were used, suggesting the need for a more detailed investigation of the different sources of fecal contamination. Area-specific threshold values for fecal indicators were estimated on a risk-basis by modelling the cumulative risk against E. coli and enterococci concentrations. To improve bathing waters assessment, we suggest considering source apportionment, locally estimating of pathogen/indicator ratios, and calculating site-specific indicators thresholds based on risk assessment.

Preliminary Data Related to the Effect of Climacostol Produced by the Freshwater Ciliate Climacostomum virens on Human Adenovirus.

The new epidemiological scenario has so far focused on the environmental circulation of human viral pathogens. Owing to the side effects of chemical disinfectants, there is an increasing need for knowledge on the use of virucidal compounds, especially those of a natural origin. Climacostol is a molecule produced by a freshwater ciliate and it exhibits activity against bacterial and fungal pathogens. We thus also speculated that there might be an effect on viral viability, which has never been tested. To evaluate such activity, we chose human adenovirus (HAdV), which is representative of waterborne viruses. We conducted experiments using HAdV serotype 5, whose titer was determined by infecting HeLa cell cultures. HAdV5 was shown to be sensitive to climacostol at a concentration of 0.0002 mg/mL, with an approximate 3 Log10 reduction when the initial titer of HAdV5 was approximately 104 and 103 TCID50/mL. These preliminary results could be an important starting point for further research aimed at improving the characterization of climacostol activity under different experimental conditions and against various viruses, including enveloped ones (i.e., the coronavirus). The production of climacostol by a protist living in fresh water also suggests a possible application in the activated sludge of wastewater treatment plants.

Making Waves: Coronavirus detection, presence and persistence in the water environment: State of the art and knowledge needs for public health.

The main route of transmission of the human coronaviruses (HCoVs), and presumably also of the new pandemic SARS-CoV-2, is via droplets and close contacts, however their fecal elimination also suggests the possible spread via water. A scientific literature search was thus carried out to highlight the current state of the art and knowledge gaps regarding coronavirus in water. Since 1978 only 22 studies have met the inclusion criteria, and considered heterogeneous purposes, detection methods and types of water. In vitro experiments have addressed the recovery efficiency of analytical methods, survival in different types of water and the removal efficiency of water treatments. Field studies have monitored coronaviruses in surface waters, sewage, slurry, and biosolids. Overall, at the lab scale, HCoVs or surrogates can survive for several days at 4 °C, however their persistence is lower compared with non-enveloped viruses and is strongly influenced by temperature and organic or microbial pollution. HCoVs have rarely been detected in field investigations, however may be due to the low recovery efficiency of the analytical methods. The scarcity of information on HCoV in the environment suggests that research is needed to understand the fate of these viruses in the water cycle.