Monitoring of SARS-CoV-2 in wastewater: what normalisation for improved understanding of epidemic trends?

SARS-CoV-2 RNA quantification in wastewater has emerged as a relevant additional means to monitor the COVID-19 pandemic. However, the concentration can be affected by black water dilution factors or movements of the sewer shed population, leading to misinterpretation of measurement results. The aim of this study was to evaluate the performance of different indicators to accurately interpret SARS-CoV-2 in wastewater. Weekly/bi-weekly measurements from three cities in France were analysed from February to September 2021. The concentrations of SARS-CoV-2 gene copies were normalised to the faecal-contributing population using simple sewage component indicators. To reduce the measurement error, a composite index was created to combine simultaneously the information carried by the simple indicators. The results showed that the regularity (mean absolute difference between observation and the smoothed curve) of the simple indicators substantially varied across sampling points. The composite index consistently showed better regularity compared to the other indicators and was associated to the lowest variation in correlation coefficient across sampling points. These findings suggest the recommendation for the use of a composite index in wastewater-based epidemiology to compensate for variability in measurement results.

Elimination of SARS-CoV-2 along wastewater and sludge treatment processes.

The Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is shed in the feces of infected people. As a consequence, genomic RNA of the virus can be detected in wastewater. Although the presence of viral RNA does not inform on the infectivity of the virus, this presence of genetic material raised the question of the effectiveness of treatment processes in reducing the virus in wastewater and sludge. In this work, treatment lines of 16 wastewater treatment plants were monitored to evaluate the removal of SARS-CoV-2 RNA in raw, processed waters and sludge, from March to May 2020. Viral RNA copies were enumerated using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) in 5 different laboratories. These laboratories participated in proficiency testing scheme and their results demonstrated the reliability and comparability of the results obtained for each one. SARS-CoV-2 RNA was found in 50.5% of the 101 influent wastewater samples characterized. Positive results were detected more frequently in those regions with a COVID-19 incidence higher than 100 cases per 100,000 inhabitants. Wastewater treatment plants (WWTPs) significantly reduced the occurrence of virus RNA along the water treatment lines. Secondary treatment effluents showed an occurrence of SARS-CoV-2 RNA in 23.3% of the samples and no positive results were found after MBR and chlorination. Non-treated sludge (from primary and secondary treatments) presented a higher occurrence of SARS-CoV-2 RNA than the corresponding water samples, demonstrating the affinity of virus particles for solids. Furthermore, SARS-CoV-2 RNA was detected in treated sludge after thickening and anaerobic digestion, whereas viral RNA was completely eliminated from sludge only when thermal hydrolysis was applied. Finally, co-analysis of SARS-CoV-2 and F-specific RNA bacteriophages was done in the same water and sludge samples in order to investigate the potential use of these bacteriophages as indicators of SARS-CoV-2 fate and reduction along the wastewater treatment.

Airborne mycotoxins in waste recycling and recovery facilities: Occupational exposure and health risk assessment.

Mycotoxins are metabolites secreted by certain types of moulds, and some of them can be harmful to health. The objective of this study was to estimate the level of exposure to airborne aflatoxin B1, ochratoxin A, gliotoxin and sterigmatocystin in waste recycling and recovery facilities. An additional goal was to assess the related health risks for workers. Targeted mycotoxins were analysed quantitatively in 94 air samples collected in five sites using ultra-performance liquid chromatography coupled with high resolution mass spectrometry. The level of exposure to mycotoxin during working day scenarios was compared to benchmark values, either health-based guidelines when available or the concentration of no toxicological concern (CoNTC) when not. Eleven per cent of samples showed quantifiable measurement results. Aflatoxin B1 and sterigmatocystin were quantified at the mechanical separation area in mechanical-biological treatment (MBT) facilities and in the materials recovery facility (MRF), but not in composting plants and composting units in MBT facilities. The levels of exposure were all below 1 ng m-3. This is the first time exposure to sterigmatocystin in waste management facilities is reported and quantified. Ochratoxin A and gliotoxin were not quantified in any of the air samples. Health risk assessment approaches did not suggest a significant threat to workers' health. These data do not suggest the need for specific prevention measures in addition to those against other airborne biological agents.

Inhalable dust as a marker of exposure to airborne biological agents in composting facilities.

OBJECTIVES: Industrial composting is associated with high levels of worker exposure to bioaerosols. Measurement of airborne microorganisms and endotoxin is complex and the related cost is high. The objective was therefore to examine whether dust measurement could be used as a marker of exposure to bioaerosols in composting facilities. METHODS: A dataset of 110 measurements carried out in eight sludge composting plants was explored. Mixed-effects models were constructed to explain between-site and within-site variability in concentration of endotoxin and culturable mesophilic bacteria, mesophilic moulds and thermophilic actinomycetes in air. Fixed-effects variables were inhalable dust concentration, the season, the outdoor/indoor location of sampling and the process area. RESULTS: The level of dust was a highly significant determinant of concentration for all biological agents. Within-site variability was always larger than between-site variability. The proportion of within-site variability explained by determinants was 68%, 65%, 56% and 60% for endotoxin, bacteria, moulds and actinomycetes, respectively. Inclusion of dust in the final model resulted in an increase of 24, 20, 12 and 17 points of percentage within-site variability, respectively. Inclusion of season resulted in an increase of 9, 12, 12 and 15 points, respectively. Within-site variability was less influenced by outdoor/indoor location and process area, except for moulds. CONCLUSION: Dust was the factor that most influenced within-site variability in endotoxin and culturable bacteria concentration. Measurement of dust can efficiently assist decision making for prevention measures against endotoxin and bacteria in sludge composting plants. Our results are not as conclusive for actinomycetes and especially for moulds.

Aspergillus fumigatus and mesophilic moulds in air in the surrounding environment downwind of non-hazardous waste landfill sites.

Non-hazardous waste landfilling has the potential to release biological agents into the air, notably mould spores. Some species, such as Aspergillus fumigatus, may be a cause of concern for at-risk nearby residents. However, air concentration in the surrounding environment of non-hazardous waste landfill sites is poorly documented. An extensive sampling programme was designed to investigate the relationship between culturable mesophilic moulds and A. fumigatus concentrations in air and distance downwind of non-hazardous waste landfill sites. On-site and off-site repeated measurements were performed at four landfill sites during cold and warm seasons. A high-flow air-sampler device was selected so as to allow peak concentration measurement. Linear mixed-effects models were used to explain variability in the concentrations in air over time and across sites, seasons, instantaneous meteorological conditions and discharged waste tonnage. Concentrations of mesophilic moulds and A. fumigatus at off-site upwind sampling locations were compared with concentrations at each of the downwind sampling locations. At the tipping face location, peak concentration reached 480,000CFUm(-3) for mesophilic moulds and 9300CFUm(-3) for A. fumigatus. Compared with upwind background levels, these concentrations were, on average, approximately 20 and 40 times higher respectively. A steep decline in the concentration of both mesophilic moulds and A. fumigatus was observed between the tipping face location and the downwind property boundary (reduction by 77% and 84% respectively), followed by a low decline leading to a 90% and 94% reduction in concentration at 200m from the property boundary and beyond. With the 200m and 500m downwind sampling point values added together, the 97.5th percentile of concentration was 6013CFUm(-3) and 87CFUm(-3) for mesophilic moulds and A. fumigatus, respectively. Other determining factors were the discharged waste tonnage, the season, instantaneous temperature and wind velocity for mesophilic mould, and instantaneous temperature for A. fumigatus. At 200m and 500 downwind from the property boundary, mesophilic moulds and A. fumigatus concentrations were still higher than the local background level. However, whilst statistically significant, this increase does not suggest an excess risk to nearby residents' health when compared with the wide range of outdoor background levels reported in literature. These findings suggest that moulds and A. fumigatus may be transported beyond 200m from the property boundary in concentrations above those found locally upwind of the landfill site. Nevertheless, for exposure assessment purposes, comparison should also be made with background levels in wider areas which are either residential or through which people travel to work for example.

Drinking water, diet, indoor air: Comparison of the contribution to environmental micropollutants exposure.

This study collated 254,441 analytical results from drinking water quality monitoring in order to compare levels of exposure of the French adult population from drinking water with that from total diet for 37 pesticides, 11 mineral elements, 11 polycyclic aromatic hydrocarbons (PAH), 6 non dioxin-like polychlorobiphenyls (NDL PCB), 5 ether polybromodiphenyl ethers (BDE), 2 perfluorinated compounds. It also compares levels of exposure from drinking water with that from inhalation of indoor air for 9 volatile organic compounds (VOC) and 3 phthalates. The vast majority of the water analysis results showed values below the limits of quantification and this comparison was primarily made on the basis of a highly pessimistic scenario consisting in considering the data below the limits of quantification as being equal to the limits of quantification. With this conservative scenario, it can be seen that tap water makes a minor but potentially non-negligible contribution for a few micropollutants, by comparison with diet and air. It also shows that exposure through drinking water remains below the toxicity reference values for these substances. Apart from a few extreme values reflecting exceptional local situations, the concentrations measured for the minority of positive samples (below the 95th percentile value) suggest a very low risk for human health. Lower limits of quantification would however be of use in better estimating the safety margin with regard to the toxicity reference values, in particular for BDE, PAH and NDL PCB.

Bioaerosol in composting facilities: occupational health risk assessment.

This research found evidence of an association between occupational exposure to bioaerosols in composting plants and health outcome occurrence in exposed workers. An occupational exposure assessment in six composting plants was performed to better characterize personal exposure levels and evaluate associated health risk in workers. Sampling results showed large ranges of concentrations of dust, bacteria, molds, and endotoxin in ambient air and in personal samples, both when driving a front-end loader and when cleaning, monitoring, and performing maintenance tasks. Mean personal exposure levels were high at 100 to more than 10,000 times higher than outdoor background levels and fully consistent with occurrence of inflammatory and allergic respiratory outcomes among workers. Engineering control, personal protection, and education and training programs for employees, health, and safety officials, and occupational physicians are being developed and implemented.

Fault tree analysis of the causes of waterborne outbreaks.

Prevention and containment of outbreaks requires examination of the contribution and interrelation of outbreak causative events. An outbreak fault tree was developed and applied to 61 enteric outbreaks related to public drinking water supplies in the EU. A mean of 3.25 causative events per outbreak were identified; each event was assigned a score based on percentage contribution per outbreak. Source and treatment system causative events often occurred concurrently (in 34 outbreaks). Distribution system causative events occurred less frequently (19 outbreaks) but were often solitary events contributing heavily towards the outbreak (a mean % score of 87.42). Livestock and rainfall in the catchment with no/inadequate filtration of water sources contributed concurrently to 11 of 31 Cryptosporidium outbreaks. Of the 23 protozoan outbreaks experiencing at least one treatment causative event, 90% of these events were filtration deficiencies; by contrast, for bacterial, viral, gastroenteritis and mixed pathogen outbreaks, 75% of treatment events were disinfection deficiencies. Roughly equal numbers of groundwater and surface water outbreaks experienced at least one treatment causative event (18 and 17 outbreaks, respectively). Retrospective analysis of multiple outbreaks of enteric disease can be used to inform outbreak investigations, facilitate corrective measures, and further develop multi-barrier approaches.