Background Clinical detection of SARS-CoV-2 RNA in stools supports the idea of wastewater-based epidemiology (WBE) as a precious tool for COVID-19 environmental surveillance. Successful detection of SARS-CoV-2 RNA in untreated wastewaters has been reported in several countries. This study investigated the presence and persistence of viral RNA in treated and untreated wastewaters in Padua, Italy. An urban experimental network of sampling sites was tested for prospective surveillance activities. Methods Seven sampling sites (i.e. wastewater pumping stations, plant inlets and outlets) were selected from the two main municipal wastewater treatment plant systems. Eleven grab samples (9 untreated, 2 treated wastewaters) were collected on 2 dates. All samples were tested at t0 for SARS-CoV-2 RNA and t1 = 24 h to investigate its persistence, at room temperature and under refrigerated conditions. Overall, 33 sub-samples were concentrated by ultrafiltration and tested for molecular detection of viral RNA with two RT-qPCR assays. Results At t0, positivity for at least one RT-qPCR assay was achieved by 4/9 untreated wastewater samples and 2/2 tertiary treated samples. A minimum SARS-CoV-2 titer of 4.8–4.9 log10 gc/L was estimated. At t1, three refrigerated subsamples were positive as well. The two RT-qPCR assays showed differential sensitivity, with the N assay detecting 90% of successful amplifications. Conclusions SARS-CoV-2 RNA was detected in untreated and treated wastewaters. Its persistence after 24 h was demonstrated in subsamples kept at 4 °C. Hospitalization data suggested an approximate WBE detection power of 1 COVID-19 case per 531 inhabitants. The possible role of WBE in COVID-19 environmental surveillance is strongly supported by our findings. WBE can also provide precious support in the decision-making process of restriction policies during the epidemic remission phase. Optimization and standardization of laboratory methods should be sought in the short term, so that results from different studies can be compared with reliability.
The airborne transmission of SARS-CoV-2, the etiologic agent of the current COVID-19 pandemic, has been hypothesized as one of the primary routes of transmission. Current data suggest a low probability of airborne transmission of the virus in open environments and a higher probability in closed ones, particularly in hospitals or quarantine facilities. However, the potential diffusion of the virus in open environments, especially using particulate matter (PM) as a transport carrier, generated concern in the exposed populations. Several authors found a correlation between the exceeding of the PM10 concentration limits in some Italian cities and the prevalence of Covid-19 cases detected in those areas. This study investigated the potential presence of SARS-COV-2 RNA on a representative series of PM samples collected in the province of Padua in Northeastern Italy during the first wave of COVID pandemic. Forty-four samples of PM2.5 and PM10 were collected between February 24 and March 9, 2020 and analyzed with RT-qPCR for SARS-CoV-2 RNA. The experimental results did not indicate the presence of SARS-CoV-2 RNA in the outdoor PMs, thus confirming the low probability of virus airborne transmission through PM.
Evaluation of hygienic aspects of thermal mud microbiology is still neglected. This study evaluates the microbiological hygiene quality of thermal muds, providing a comprehensive assessment of the whole mud cultivation chain. Maturing mud, peloid and used mud samples were collected twice in a year from 30 SPAs of the Euganean Thermal District, NE Italy. Samples were processed with an ad hoc laboratory method. The following indicator parameters were assessed: Total Count at 22, 37 and 55 °C; total coliforms; Escherichia coli; enterococci; Staphylococcus aureus; Pseudomonas aeruginosa; sulfite-reducing clostridia; dermatophytes. Statistical significance of differences between the two sampling campaigns and correlation between temperature and indicator parameters were evaluated. One-hundred eighty samples were analyzed. Widespread presence of environmental species was found, as well as hints of possible microorganism transfer from the patient’s skin to the mud. Proper setting of thermal water temperature resulted critical, in terms of hygienic quality. Although optimal maturation should be granted (thermal water at 30–42 °C), a pasteurization step at 60–65 °C is strongly recommended to sanitize peloids before pelotherapy. Facilities re-using thermal muds should also implement a regeneration step at ≥65 °C. Core evaluation of thermal mud hygienic quality could encompass the following guidelines: absence (i.e., 0 colony forming units (CFU)/g) of E. coli, P. aeruginosa, S. aureus and dermatophytes.
Analysis of atmospheric particulate matter (PM) has been proposed for the environmental surveillance of SARS-CoV-2. The aim of this study was to increase the current knowledge about the occurrence of SARS-CoV-2 in atmospheric PM, introduce a dedicated sampling method, and perform a simultaneous assessment of human seasonal coronavirus 229E. Thirty-two PM samples were collected on quartz fiber filters and six on Teflon using a low- and high-volumetric rate sampler, respectively, adopting a novel procedure for optimized virus detection. Sampling was performed at different sites in the Venice area (Italy) between 21 February and 8 March 2020 (n = 16) and between 27 October and 25 November 2020 (n = 22). A total of 14 samples were positive for Coronavirus 229E, 11 of which were collected in October–November 2020 (11/22; positivity rate 50%) and 3 in February–March 2020 (3/16 samples, 19%). A total of 24 samples (63%) were positive for SARS-CoV-2. Most of the positive filters were collected in October–November 2020 (19/22; positivity rate, 86%), whereas the remaining five were collected in February–March 2020 at two distinct sites (5/16, 31%). These findings suggest that outdoor PM analysis could be a promising tool for environmental surveillance. The results report a low concentration of SARS-CoV-2 in outdoor air, supporting a scarce contribution to the spread of infection.
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