Essential oil mediated synthesis and application of highly stable copper nanoparticles as coatings on textiles and surfaces for rapid and sustained disinfection of microorganisms.

Rampant pathogenesis induced by communicable microbes has necessitated development of technologies for rapid and sustained disinfection of surfaces. Copper nanoparticles (CuNPs) have been widely reported for their antimicrobial properties. However, nanostructured copper is prone to oxidative dissolution in the oil phase limiting its sustained use on surfaces and coatings. The current study reports a systematic investigation of a simple synthesis protocol using fatty acid stabilizers (particularly essential oils) for synthesis of copper nanoparticles in the oil phase. Of the various formulations synthesized, rosemary oil stabilized copper nanoparticles (RMO CuNPs) were noted to have the best inactivation kinetics and were also most stable. Upon morphological characterization by TEM and EELS, these were found to be monodispersed (φ5-8 nm) with copper coexisting in all three oxidation states on the surface of the nanoparticles. The nanoparticles were drop cast on woven fabric of around 500 threads per inch and exposed to gram positive bacteria (Staphylococcus aureus), gram negative bacteria (Escherichia coliandPseudomonas aeruginosa), enveloped RNA virus (phi6), non-enveloped RNA virus (MS2) and non-enveloped DNA virus (T4) to encompass the commonly encountered groups of pathogens. It was possible to completely disinfect 107copies of all microorganisms within 40 min of exposure. Further, this formulation was incorporated with polyurethane as thinners and used to coat non-woven fabrics. These also exhibited antimicrobial properties. Sustained disinfection with less than 9% cumulative copper loss for upto 14 washes with soap water was observed while the antioxidant activity was also preserved. Based on the studies conducted, RMO CuNP in oil phase was found to have excellent potential of integration on surface coatings, paints and polymers for rapid and sustained disinfection of microbes on surfaces.

Corrigendum: Wastewater-based surveillance identifies start to the pediatric respiratory syncytial virus season in two cities in Ontario, Canada.

[This corrects the article DOI: 10.3389/fpubh.2023.1261165.].

Impact of coagulation on SARS-CoV-2 and PMMoV viral signal in wastewater solids.

Wastewater surveillance (WWS) of SARS-CoV-2 has become a crucial tool for monitoring COVID-19 cases and outbreaks. Previous studies have indicated that SARS-CoV-2 RNA measurement from testing solid-rich primary sludge yields better sensitivity compared to testing wastewater influent. Furthermore, measurement of pepper mild mottle virus (PMMoV) signal in wastewater allows for precise normalization of SARS-CoV-2 viral signal based on solid content, enhancing disease prevalence tracking. However, despite the widespread adoption of WWS, a knowledge gap remains regarding the impact of ferric sulfate coagulation, commonly used in enhanced primary clarification, the initial stage of wastewater treatment where solids are sedimented and removed, on SARS-CoV-2 and PMMoV quantification in wastewater-based epidemiology. This study examines the effects of ferric sulfate addition, along with the associated pH reduction, on the measurement of SARS-CoV-2 and PMMoV viral measurements in wastewater primary clarified sludge through jar testing. Results show that the addition of Fe3+ concentrations in the conventional 0 to 60 mg/L range caused no effect on SARS-CoV-2 N1 and N2 gene region measurements in wastewater solids. However, elevated Fe3+ concentrations were shown to be associated with a statistically significant increase in PMMoV viral measurements in wastewater solids, which consequently resulted in the underestimation of PMMoV-normalized SARS-CoV-2 viral signal measurements (N1 and N2 copies/copies of PMMoV). The observed pH reduction from coagulant addition did not contribute to the increased PMMoV measurements, suggesting that this phenomenon arises from the partitioning of PMMoV viral particles into wastewater solids.

Wastewater-based surveillance identifies start to the pediatric respiratory syncytial virus season in two cities in Ontario, Canada.

Introduction: Detection of community respiratory syncytial virus (RSV) infections informs the timing of immunoprophylaxis programs and hospital preparedness for surging pediatric volumes. In many jurisdictions, this relies upon RSV clinical test positivity and hospitalization (RSVH) trends, which are lagging indicators. Wastewater-based surveillance (WBS) may be a novel strategy to accurately identify the start of the RSV season and guide immunoprophylaxis administration and hospital preparedness. Methods: We compared citywide wastewater samples and pediatric RSVH in Ottawa and Hamilton between August 1, 2022, and March 5, 2023. 24-h composite wastewater samples were collected daily and 5 days a week at the wastewater treatment facilities in Ottawa and Hamilton, Ontario, Canada, respectively. RSV WBS samples were analyzed in real-time for RSV by RT-qPCR. Results: RSV WBS measurements in both Ottawa and Hamilton showed a lead time of 12 days when comparing the WBS data set to pediatric RSVH data set (Spearman's ρ = 0.90). WBS identify early RSV community transmission and declared the start of the RSV season 36 and 12 days in advance of the provincial RSV season start (October 31) for the city of Ottawa and Hamilton, respectively. The differing RSV start dates in the two cities is likely associated with geographical and regional variation in the incidence of RSV between the cities. Discussion: Quantifying RSV in municipal wastewater forecasted a 12-day lead time of the pediatric RSVH surge and an earlier season start date compared to the provincial start date. These findings suggest an important role for RSV WBS to inform regional health system preparedness, reduce RSV burden, and understand variations in community-related illness as novel RSV vaccines and monoclonal antibodies become available.

Rapidly developed, optimized, and applied wastewater surveillance system for real-time monitoring of low-incidence, high-impact MPOX outbreak.

Recent MPOX viral resurgences have mobilized public health agencies around the world. Recognizing the significant risk of MPOX outbreaks, large-scale human testing, and immunization campaigns have been initiated by local, national, and global public health authorities. Recently, traditional clinical surveillance campaigns for MPOX have been complemented with wastewater surveillance (WWS), building on the effectiveness of existing wastewater programs that were built to monitor SARS-CoV-2 and recently expanded to include influenza and respiratory syncytial virus surveillance in wastewaters. In the present study, we demonstrate and further support the finding that MPOX viral fragments agglomerate in the wastewater solids fraction. Furthermore, this study demonstrates that the current, most commonly used MPOX assays are equally effective at detecting low titers of MPOX viral signal in wastewaters. Finally, MPOX WWS is shown to be more effective at passively tracking outbreaks and/or resurgences of the disease than clinical testing alone in smaller communities with low human clinical case counts of MPOX.

Laser-Induced Graphene Capacitive Killing of Bacteria.

Laser-induced graphene (LIG) is a method of generating a foam-like conformal carbon layer of porous graphene on many types of carbon-based surfaces. This electrically conductive material has been shown to be useful in many applications including environmental technology and includes low fouling and antimicrobial surfaces and can address persistent environmental challenges spawned by bacterial and viral contaminates. Here, we show that a single film of LIG stores charge when an electrical current is applied and dissipates charge when the current is stopped, which results in electricidal surface antibacterial potency. The amount of accumulated and dissipated charge on a single strip of LIG was quantified with an electrometer by generating LIG on both sides of a nonconducting polyimide film, which showed up to 65 pC of charge when the distance between the surfaces was 94 µm corresponding to an areal capacitance of 1.63 pF/cm2. We further corroborate the stored charge decay of a single LIG strip with bacteria death via direct electrical contact. Antimicrobial rates decreased with the same monotonic pattern as the loss of charge from the LIG film (i.e., AR ∼ 97% 0 s after voltage source disconnection vs AR ∼ 21% 90 s after disconnection) showing bacterial death as a function of delayed LIG exposure time after applied voltage disconnection. In terms of energy efficiency, this translates to an increased bacteria potency of ∼170% for the equivalent energy costs as that previously estimated. Finally, we present a mechanistic explanation for the capacitive behavior and the electricidal effects for a single plate of LIG.

Municipal and neighbourhood level wastewater surveillance and subtyping of an influenza virus outbreak.

Recurrent influenza epidemics and pandemic potential are significant risks to global health. Public health authorities use clinical surveillance to locate and monitor influenza and influenza-like cases and outbreaks to mitigate hospitalizations and deaths. Currently, global integration of clinical surveillance is the only reliable method for reporting influenza types and subtypes to warn of emergent pandemic strains. The utility of wastewater surveillance (WWS) during the COVID-19 pandemic as a less resource intensive replacement or complement for clinical surveillance has been predicated on analyzing viral fragments in wastewater. We show here that influenza virus targets are stable in wastewater and partitions favorably to the solids fraction. By quantifying, typing, and subtyping the virus in municipal wastewater and primary sludge during a community outbreak, we forecasted a citywide flu outbreak with a 17-day lead time and provided population-level viral subtyping in near real-time to show the feasibility of influenza virus WWS at the municipal and neighbourhood levels in near real time using minimal resources and infrastructure.

Occurrence of organic micropollutants in municipal landfill leachate and its effective treatment by advanced oxidation processes.

Landfilling is the most prominently adopted disposal technique for managing municipal solid waste across the globe. However, the main drawback associated with this method is the generation of leachate from the landfill site. Leachate, a highly concentrated liquid consisting of both organic and inorganic components arises environmental issues as it contaminates the nearby aquifers. Landfill leachate treatment by conventional methods is not preferred as the treatment methods are not much effective to remove these pollutants. Advanced oxidation processes (AOPs) based on both hydroxyl and sulfate radicals could be a promising method to remove the micropollutants completely or convert them to non-toxic compounds. The current review focuses on the occurrence of micropollutants in landfill leachate, their detection methods and removal from landfill leachate using AOPs. Pharmaceuticals and personal care products occur in the range of 10-1 to more than 100 µg L-1 whereas phthalates were found below the detectable limit to 384 µg L-1, pesticides in the order of 10-1 µg L-1 and polyaromatic hydrocarbons occur in concentration from 10-2 to 114.7 µg L-1. Solid-phase extraction is the most preferred method for extracting micropollutants from leachate and liquid chromatography (LC) - mass spectrophotometer (MS) for detecting the micropollutants. Limited studies have been focused on AOPs as a potential method for the degradation of micropollutants in landfill leachate. The potential of Fenton based techniques, electrochemical AOPs and ozonation are investigated for the removal of micropollutants from leachate whereas the applicability of photocatalysis for the removal of a wide variety of micropollutants from leachate needs in-depth studies.

Low-Voltage Bacterial and Viral Killing Using Laser-Induced Graphene-Coated Non-woven Air Filters.

Laser-induced graphene (LIG) is uniquely positioned to advance applications in which electrically conductive carbon coatings are required. Recently, the antifouling, antiviral, and antibacterial properties of LIG have been proven in both air and water filtration applications. For example, an unsupported LIG based filter (pore size: ∼0.3 µm) demonstrated exceptional air filtration properties, while its joule heating effects successfully sterilized and removed unwanted biological components in air despite persisting challenges such as pressure drop, energy consumption, and lack of mechanical robustness. Here, we developed a polyimide (PI) non-woven supported LIG air filter with negligible pressure drop changes compared to the non-woven support material and showed that low electrical current density inactivates aerosolized bacteria. A current density of 4.5 mA/cm2 did not cause significant joule heating, and 97.2% bacterial removal was obtained. The low-voltage antibacterial mechanism was elucidated using bacterial inhibition experiments on a titanium surface and on an LIG surface fabricated on dense PI films. Complete sterilization was obtained using current densities of ∼8 mA/cm2 applied for 2 min or ∼ 6 mA/cm2 for 10 min upon the dense PI-LIG surface. Lastly, >98% bacterial removal was observed using a low-resistance LIG-coated non-woven polyimide air filter at 5 V. However, only very low voltages (∼0.3 V) were needed to remove ∼99% Pseudomonas aeruginosa bacteria and 100% of T4 virus when the LIG-coated filters were hybridized with a stainless steel mesh. Our results show that low current density levels at very low voltages are sufficient for substantial bacterial and viral inactivation, and that these principles might be effectively used in a wide number of air filtration applications such as air conditioners or other ventilation systems, which might limit the spread of infectious particles in hospitals, homes, workplaces, and the transportation industry.

Polyphenol stabilized copper nanoparticle formulations for rapid disinfection of bacteria and virus on diverse surfaces.

Rapid and sustained disinfection of surfaces is necessary to check the spread of pathogenic microbes. The current study proposes a method of synthesis and use of copper nanoparticles (CuNPs) for contact disinfection of pathogenic microorganisms. Polyphenol stabilized CuNPs were synthesized by successive reductive disassembly and reassembly of copper phenolic complexes. Morphological and compositional characterization by transmission electron microscope (TEM), selected area diffraction and electron energy loss spectroscopy revealed monodispersed spherical (ϕ5-8 nm) CuNPs with coexisting Cu, Cu(I) and Cu (II) phases. Various commercial grade porous and non-porous substrates, such as, glass, stainless steel, cloth, plastic and silk were coated with the nanoparticles. Complete disinfection of 107copies of surrogate enveloped and non-enveloped viruses: bacteriophage MS2, SUSP2, phi6; and gram negative as well as gram positive bacteria:Escherichia coliandStaphylococcus aureuswas achieved on most substrates within minutes. Structural cell damage was further analytically confirmed by TEM. The formulation was well retained on woven cloth surfaces even after repeated washing, thereby revealing its promising potential for use in biosafe clothing. In the face of the current pandemic, the nanomaterials developed are also of commercial utility as an eco-friendly, mass producible alternative to bleach and alcohol based public space sanitizers used today.

Development and evaluation of DEAE silica gel columns for simultaneous concentration of coliphages and rotavirus from natural water samples.

Enteric viruses are commonly present in water bodies in regions with poor sanitation. Although the occurrence of these viruses poses a health risk they are difficult to quantify due to their low concentration and they may remain undetected in the absence of adequate preconcentration. The present study reports the synthesis and utilization of DEAE silica gel (DSiG) as an adsorbent for virus concentration. Two coliphages, MS2 and SUSP2, and an enteric virus, rotavirus A (RVA) were chosen for examining the preconcentration efficiency of DSiG columns. Studies conducted at a low flow rate of 5 mL/min yielded good removal of viruses through adsorption. Studies at a higher flow rate of 50 mL/min followed by elution with optimized eluents yielded a high recovery of MS2 and RVA even when they were present at low concentration (0.01 copy/mL). The eluent Na(1.5 M)-Tw(2%)-G3X (glycine 3X broth, 1.5 M NaCl, 2% Tween, pH 10.2) showed maximum elution of RVA and MS2. Optimal SUSP2 recovery was observed on employing an eluent composed of 1.5 M NaCl, 3% Tween, 0.05 M KH2PO4 at pH 9.2. Subsequently, both the eluents were successively applied for elution of the adsorbed viruses. This method was applied for virus preconcentration from lake water in the monsoon and winter seasons. The DSiG column could achieve adequate preconcentration for all the three viruses, i.e., SUSP2, MS2, and RVA, even when they were present at very low concentration and the recovery achieved was comparable to that achieved with ultracentrifugation while the processing time required for handling large volumes of water was considerably lower.

The novel SARS-CoV-2 pandemic: Possible environmental transmission, detection, persistence and fate during wastewater and water treatment.

The contagious SARS-CoV-2 virus, responsible for COVID-19 disease, has infected over 27 million people across the globe within a few months. While literature on SARS-CoV-2 indicates that its transmission may occur predominantly via aerosolization of virus-laden droplets, the possibility of alternate routes of transmission and/or reinfection via the environment requires considerable scientific attention. This review aims to collate information on possible transmission routes of this virus, to ascertain its fate in the environment. Concomitant with the presence of SARS-CoV-2 viral RNA in faeces and saliva of infected patients, studies also indicated its occurrence in raw wastewater, primary sludge and river water. Therefore sewerage system could be a possible route of virus outbreak, a possible tool to assess viral community spread and future surveillance technique. Hence, this review looked into detection, occurrence and fate of SARS-CoV-2 during primary, secondary, and tertiary wastewater and water treatment processes based on published literature on SARS-CoV and other enveloped viruses. The review also highlights the need for focused research on occurrence and fate of SARS-CoV-2 in various environmental matrices. Utilization of this information in environmental transmission models developed for other enveloped and enteric viruses can facilitate risk assessment studies. Preliminary research efforts with SARS-CoV-2 and established scientific reports on other coronaviruses indicate that the threat of virus transmission from the aquatic environment may be currently non-existent. However, the presence of viral RNA in wastewater provides an early warning that highlights the need for effective sewage treatment to prevent a future outbreak of SARS-CoV-2.