Variability in the recovery of a virus concentration procedure in water: Implications for QMRA

Petterson, Susan; Grøndahl-Rosado, Ricardo; Nilsen, Vegard; Myrmel, Mette; Robertson, Lucy J.

doi:10.1016/j.watres.2015.09.006

Cited by 53 publications

(40 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, an average recovery of 1% and 2% from mussels and oysters, respectively, was achieved. Therefore, the corrected (to account for extraction efficacy) NoV values would be of a greater titre than uncorrected values, with important considerations for risk assessment and proposed shellfish standards (Petterson et al 2015). …”

Section: Detection and Quantification Of Novmentioning

confidence: 99%

See 1 more Smart Citation

Critical Review on the Public Health Impact of Norovirus Contamination in Shellfish and the Environment: A UK Perspective

et al. 2017

View full text Add to dashboard Cite

We review the risk of norovirus (NoV) infection to the human population from consumption of contaminated shellfish. From a UK perspective, risk is apportioned for different vectors of NoV infection within the population. NoV spreads mainly by person-to-person contact or via unsanitary food handling. NoV also enters the coastal zone via wastewater discharges resulting in contamination of shellfish waters. Typically, NoV persists in the marine environment for several days, with its presence strongly linked to human population density, wastewater discharge rate, and efficacy of wastewater treatment. Shellfish bioaccumulate NoV and current post-harvest depuration is inefficient in its removal. While NoV can be inactivated by cooking (e.g. mussels), consumption of contaminated raw shellfish (e.g. oysters) represents a risk to human health. Consumption of contaminated food accounts for 3–11% of NoV cases in the UK (~74,000 cases/year), of which 16% are attributable to oyster consumption (11,800 cases/year). However, environmental and human factors influencing NoV infectivity remain poorly understood. Lack of standard methods for accurate quantification of infective and non-infective (damaged) NoV particles represent a major barrier, hampering identification of an appropriate lower NoV contamination limit for shellfish. Future management strategies may include shellfish quality assessment (at point of harvest or at point of supply) or harvesting controls. However, poor understanding of NoV inactivation in shellfish and the environment currently limits accurate apportionment and risk assessment for NoV and hence the identification of appropriate shellfish or environmental quality standards.

show abstract

Section: Detection and Quantification Of Novmentioning

confidence: 99%

“…However, in the absence of this information, a hybrid solution is possible, whereby an E. coli proxy is used for NoV risk (Petterson et al 2015). This proxy could be linked to target NoV standards for harvest, and could be adjusted accordingly on the advent of new information.…”

Section: Regulating Nov In Shellfish Waters and Shellfish: Current Prmentioning

confidence: 99%

Critical Review on the Public Health Impact of Norovirus Contamination in Shellfish and the Environment: A UK Perspective

et al. 2017

View full text Add to dashboard Cite

show abstract

“…Recoveries of viral RNA from sediments range from 0.09 to 11% for direct extraction and RT-qPCR, with improved extraction efficiency when applying indirect elution-concentration approaches (Table 7). There are numerous reports of inhibition of PCR assays by organic matter (e.g., humic acids) often found in environmental samples (Meschke and Sobsey, 1998; Rock et al, 2010) and extraction and enumeration methods strongly influence estimates of viral abundance in sediments (Williamson et al, 2013), which can greatly influence attributed risk in pathogenic strains (Petterson et al, 2015). Recently, methods have been applied to overcome this; for example, Carreira et al (2015) found that a combination of EDTA in addition to probe sonication and enzymatic pre-treatments resulted in 4.5 fold increase in viral recovery from sediments.…”

Section: Fate and Behavior Of Fecally Derived Viruses In Sedimentsmentioning

confidence: 99%

Abundance and Distribution of Enteric Bacteria and Viruses in Coastal and Estuarine Sediments—a Review

et al. 2016

View full text Add to dashboard Cite

The long term survival of fecal indicator organisms (FIOs) and human pathogenic microorganisms in sediments is important from a water quality, human health and ecological perspective. Typically, both bacteria and viruses strongly associate with particulate matter present in freshwater, estuarine and marine environments. This association tends to be stronger in finer textured sediments and is strongly influenced by the type and quantity of clay minerals and organic matter present. Binding to particle surfaces promotes the persistence of bacteria in the environment by offering physical and chemical protection from biotic and abiotic stresses. How bacterial and viral viability and pathogenicity is influenced by surface attachment requires further study. Typically, long-term association with surfaces including sediments induces bacteria to enter a viable-but-non-culturable (VBNC) state. Inherent methodological challenges of quantifying VBNC bacteria may lead to the frequent under-reporting of their abundance in sediments. The implications of this in a quantitative risk assessment context remain unclear. Similarly, sediments can harbor significant amounts of enteric viruses, however, the factors regulating their persistence remains poorly understood. Quantification of viruses in sediment remains problematic due to our poor ability to recover intact viral particles from sediment surfaces (typically <10%), our inability to distinguish between infective and damaged (non-infective) viral particles, aggregation of viral particles, and inhibition during qPCR. This suggests that the true viral titre in sediments may be being vastly underestimated. In turn, this is limiting our ability to understand the fate and transport of viruses in sediments. Model systems (e.g., human cell culture) are also lacking for some key viruses, preventing our ability to evaluate the infectivity of viruses recovered from sediments (e.g., norovirus). The release of particle-bound bacteria and viruses into the water column during sediment resuspension also represents a risk to water quality. In conclusion, our poor process level understanding of viral/bacterial-sediment interactions combined with methodological challenges is limiting the accurate source apportionment and quantitative microbial risk assessment for pathogenic organisms associated with sediments in aquatic environments.

show abstract

“…The cell culture assays strongly indicate that chlorine and temperature treatment led to a complete loss of adenovirus infectivity (≥4.34 log10; see Figure 1A,C) and UV to a large loss (3.46 log10; Figure 1B). These results demonstrate that relying on qPCR alone would overestimate the amount of infectious HAdV, with reduction rates of merely 0.07 log10 after inactivation by UV (HAdV-Hernroth) and 1.07 log10 by Chlorine (HAdV-Heim), thus resulting in misleading quantitative microbial risk assessment (QMRA) [26,71]. Utilization of PMA ci-qPCR led to a signal reduction that corresponded in part with cell culture (temperature, HAdV-Heim).…”

Section: Resultsmentioning

confidence: 99%

Capsid Integrity qPCR—An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

Leifels

Shoults

Wiedemeyer

et al. 2019

Water

View full text Add to dashboard Cite

Recreational, reclaimed and drinking source waters worldwide are under increasing anthropogenic pressure, and often contain waterborne enteric bacterial, protozoan, and viral pathogens originating from non-point source fecal contamination. Recently, the capsid integrity (ci)-qPCR, utilizing the azo-dyes propidium monoazide (PMA) or ethidium monoazide (EMA), has been shown to reduce false-positive signals under laboratory conditions as well as in food safety applications, thus improving the qPCR estimation of virions of public health significance. The compatibility of two widely used human adenovirus (HAdV) qPCR protocols was evaluated with the addition of a PMA/EMA pretreatment using a range of spiked and environmental samples. Stock suspensions of HAdV were inactivated using heat, UV, and chlorine before being quantified by cell culture, qPCR, and ci-qPCR. Apparent inactivation of virions was detected for heat and chlorine treated HAdV while there was no significant difference between ci-qPCR and qPCR protocols after disinfection by UV. In a follow-up comparative analysis under more complex matrix conditions, 51 surface and 24 wastewater samples pre/post UV treatment were assessed for enteric waterborne HAdV to evaluate the ability of ci-qPCR to reduce the number of false-positive results when compared to conventional qPCR and cell culture. Azo-dye pretreatment of non-UV inactivated samples was shown to improve the ability of molecular HAdV quantification by reducing signals from virions with an accessible genome, thereby increasing the relevance of qPCR results for public health purposes, particularly suited to resource-limited low and middle-income settings.

show abstract

Variability in the recovery of a virus concentration procedure in water: Implications for QMRA

Cited by 53 publications

References 32 publications

Critical Review on the Public Health Impact of Norovirus Contamination in Shellfish and the Environment: A UK Perspective

Critical Review on the Public Health Impact of Norovirus Contamination in Shellfish and the Environment: A UK Perspective

Abundance and Distribution of Enteric Bacteria and Viruses in Coastal and Estuarine Sediments—a Review

Capsid Integrity qPCR—An Azo-Dye Based and Culture-Independent Approach to Estimate Adenovirus Infectivity after Disinfection and in the Aquatic Environment

Contact Info

Product

Resources

About