Highlights Surface and air samples from buses and subway trains were PCR-tested for SARS-CoV-2. Evidence for fragments of viral RNA was found in 30 out of 82 samples. The potential infectivity of these samples however is considered to be extremely low. The data emphasise the importance of disinfection and hygiene in public transport. Modelling in-bus infection probability shows forced ventilation greatly reduces risk.

SARS-CoV-2 was detected in Barcelona sewage long before the declaration of the first COVID-19 case, indicating that the infection was present in the population before the first imported case was reported. Sentinel surveillance of SARS-CoV-2 in wastewater would enable adoption of immediate measures in the event of future COVID-19 waves.

Two large wastewater treatment plants (WWTP), covering around 2.7 M inhabitants, which represents around 85% of the metropolitan area of Barcelona, were sampled before, during and after the implementation of a complete lockdown. Five one-step RT-qPCR assays, targeting the polymerase (IP2 and IP4), the envelope E and the nucleoprotein (N1 and N2) genome regions, were employed for SARS-CoV-2 RNA detection in 24-h composite wastewater samples concentrated by polyethylene glycol (PEG) precipitation. SARS-CoV-2 was detected in a sewage sample collected 41 days ahead of the declaration of the first COVID-19 case. The evolution of SARS-CoV-2 genome copies in wastewater evidenced the validity of water-based epidemiology to anticipate COVID-19 outbreaks, to evaluate the impact of control measures and even to estimate the burden of shedders, including presymptomatic, asymptomatic, symptomatic and undiagnosed cases. For this latter objective, a model was applied for the estimation of the total number of shedders, evidencing a high proportion of asymptomatic infected individuals. In this way, an infection prevalence of 2.0-6.5% was figured. On the other hand, a proportion of around 0.12% and 0.09% of the total population was determined to be required for positive detection in the two WWTPs. At the end of the lockdown, SARS-CoV-2 RNA apparently disappeared in the WWTPs but could still be detected in grab samples from four urban sewers. Sewer monitoring allowed for location of specific hot spots of COVID-19, enabling the rapid adoption of appropriate mitigation measures. IMPORTANCE Water-based epidemiology (WBE) is a valuable early warning tool for tracking the circulation of the virus among the population, including not only symptomatic patients, but also asymptomatic, presymptomatic and misdiagnosed carriers, which represent a high proportion of the infected population. In the specific case of Barcelona, wastewater surveillance anticipated several weeks not only the original COVID-19 pandemic wave, but also the onset of the second wave. In addition, SARS-CoV-2 occurrence in wastewater evidenced the efficacy of the adopted lockdown measures on the circulation of the virus.

No abstract

This work aims at comparing the ability of two kinds of electrochemical technologies, namely electrocoagulation (EC) and electro-Fenton (EF), to disinfect primary and secondary effluents from municipal wastewater treatment plants. Heterotrophic bacteria, Escherichia coli, enterococci, Clostridium perfringens spores, somatic coliphages and eukaryotes (amoebae, flagellates, ciliates and metazoa) were tested as indicator microorganisms. EC with an Fe/Fe cell at 200 A m and natural pH allowed >5 log unit removal of E. coli and final concentration below 1 bacteria mL of coliphages and eukaryotes from both effluents in ca. 60 min, whereas heterotrophic bacteria, enterococci and spores were more resistant. A larger removal was obtained for the primary effluent, probably because the flocs remove higher amount of total organic carbon (TOC), entrapping more easily the microbiota. EF with a boron-doped diamond (BDD) anode and an air-diffusion cathode that produces HO on site was first performed at pH 3.0, with large or even total inactivation of microorganisms within 30 min. A more effective microorganism removal was attained as compared to EC thanks to OH formed from Fenton's reaction. A quicker disinfection was observed for the secondary effluent owing to its lower TOC content, allowing the attack of greater quantities of electrogenerated oxidants on microorganisms. Wastewater disinfection by EF was also feasible at natural pH (∼7), showing similar abatement of active microorganisms as a result of the synergistic action of generated oxidants like active chlorine and coagulation with iron hydroxides. A sequential EC/EF treatment (30 min each) was more effective for a combined decontamination and disinfection of urban wastewater.

Human norovirus (HuNoV) is a foremost cause of domestically acquired foodborne acute gastroenteritis and outbreaks. Despite industrial efforts to control HuNoV contamination of foods, its prevalence in foodstuffs at retail is significant. HuNoV infections are often associated with the consumption of contaminated produce, including ready-to-eat (RTE) salads. Decontamination of produce by washing with disinfectants is a consumer habit which could significantly contribute to mitigate the risk of infection. The aim of our study was to measure the effectiveness of chemical sanitizers in inactivating genogroup I and II HuNoV strains on mixed salads using a propidium monoazide (PMAxx)-viability RTqPCR assay. Addition of sodium hypochlorite, peracetic acid, or chlorine dioxide significantly enhanced viral removal as compared with water alone. Peracetic acid provided the highest effectiveness, with log10 reductions on virus levels of 3.66 ± 0.40 and 3.33 ± 0.19 for genogroup I and II, respectively. Chlorine dioxide showed lower disinfection efficiency. Our results provide information useful to the food industry and final consumers for improving the microbiological safety of fresh products in relation to foodborne viruses.

Norovirus infections are a leading cause of acute gastroenteritis outbreaks worldwide and across all age groups, with two main genogroups (GI and GII) infecting humans. The aim of our study was to investigate the occurrence of norovirus in saliva samples from individuals involved in outbreaks of acute gastroenteritis in closed and semiclosed institutions, and its relationship with the virus strain, virus shedding in stool, the occurrence of symptoms, age, and the secretor status of the individual. Epidemiological and clinical information was gathered from norovirus outbreaks occurring in Catalonia, Spain during 2017–2018, and stool and saliva samples were collected from affected and exposed resident individuals and workers. A total of 347 saliva specimens from 25 outbreaks were analyzed. Further, 84% of individuals also provided a paired stool sample. For GII infections, norovirus was detected in 17.9% of saliva samples from symptomatic cases and 5.2% of asymptomatic individuals. Positivity in saliva occurred in both secretors and nonsecretors. None of the individuals infected by norovirus GI was positive for the virus in saliva. Saliva positivity did not correlate with any of the studied symptoms but did correlate with age ≥ 65 years old. Individuals who were positive in saliva showed higher levels of virus shedding in stool. Mean viral load in positive saliva was 3.16 ± 1.08 log10 genome copies/mL, and the predominance of encapsidated genomes was confirmed by propidium monoazide (PMA)xx-viability RTqPCR assay. The detection of norovirus in saliva raises the possibility of oral-to-oral norovirus transmission during the symptomatic phase and, although to a lesser extent, even in cases of asymptomatic infections.

During a waterborne outbreak of norovirus in Spain, we estimated 50% illness doses for a group of exposed (secretor) persons to be 556 (95% CI 319–957) genome copies/day for norovirus GI and 2,934 (95% CI 1,683–5,044) genome copies/day for norovirus GII. Use of a propidium monoazide viability assay reduced these values.