Detection of enteric viruses in sewage sludge and treated wastewater effluent

2010 ◽  
Vol 61 (2) ◽  
pp. 537-544 ◽  
Author(s):  
A. D. Schlindwein ◽  
C. Rigotto ◽  
C. M. O. Simões ◽  
C. R. M. Barardi

Sewage sludge and treated wastewater when contaminated with enteric virus and discharged into the environment, could pose a human health risk. The aim of study was to verify the presence and viability of enteric viruses in sewage sludge and treated wastewater at a local sewage plant in Florianopolis city, Brazil. Sewage sludge was concentrated by organic flocculation and polyethylene glycol precipitation and wastewater by electronegative membrane filtration and ultrafiltration by Centriprep Concentrator. Adenovirus (AdV), hepatitis A virus (HAV), and Rotavirus (RV) were examined for all samples for 12 months and Poliovirus (PV) was also tested for in sewage sludge samples. AdV was the most prevalent in both kind of samples, followed by RV, PV (in sludge) and HAV. Viral viability by cell culture (ICC-PCR) was: AdV: 100%, HAV: 16.7%, PV: 91.7%, RV: 25% in sludge and AdV: 66.6%, HAV: 66.6% and RV: 0% in wastewater. IFA for AdV in sludge ranged from 70 to 300 FFU/ml. QPCR for AdV ranged from 4.6 × 104 to 1.2 × 106 and from 50 to 1.3 × 104 gc/ml in sludge and wastewater, respectively. HAV quantification in sludge ranged from 3.1 × 102 to 5.4 × 102 gc/ml. In conclusion, it was possible to correlate presence and viability of enteric viruses in the environmental samples analyzed.

2007 ◽  
Vol 73 (14) ◽  
pp. 4425-4428 ◽  
Author(s):  
Charles P. Gerba ◽  
Denise Kennedy

ABSTRACT This study was conducted to determine whether enteric viruses (adenovirus, rotavirus, and hepatitis A virus) added to cotton cloth swatches survive the wash cycle, the rinse cycle, and a 28-min permanent press drying cycle as commonly practiced in households in the United States. Detergent with and without bleach (sodium hypochlorite) was added to washing machines containing sterile and virus-inoculated 58-cm2 swatches, 3.2 kg of cotton T-shirts and underwear, and a soiled pillowcase designed to simulate the conditions (pH, organic load, etc.) encountered in soiled laundry. The most important factors for the reduction of virus in laundry were passage through the drying cycle and the addition of sodium hypochlorite. Washing with detergent alone was not found to be effective for the removal or inactivation of enteric viruses, as significant concentrations of virus were found on the swatches (reductions of 92 to 99%). It was also demonstrated that viruses are readily transferred from contaminated cloths to uncontaminated clothes. The use of sodium hypochlorite reduced the number of infectious viruses on the swatches after washing and drying by at least 99.99%. Laundering practices in common use in the United States do not eliminate enteric and respiratory viruses from clothes. The use of bleach can further reduce the numbers of enteric viruses in laundry.


2019 ◽  
Vol 17 (5) ◽  
pp. 670-682 ◽  
Author(s):  
Albert Simhon ◽  
Vince Pileggi ◽  
Cecily A. Flemming ◽  
José R. Bicudo ◽  
George Lai ◽  
...  

Abstract In Ontario, Canada, information is lacking on chlorine and ultraviolet (UV) light disinfection performance against enteric viruses in wastewater. We enumerated enteroviruses and noroviruses, coliphages, and Escherichia coli per USEPA methods 1615, 1602, and membrane filtration, respectively, in pre- and post-disinfection effluent at five wastewater treatment plants (WWTPs), with full-year monthly sampling, and calculated log10 reductions (LRs) while WWTPs complied with their monthly geometric mean limit of 200 E. coli/100 mL. Modeling of densities by left-censored estimation and Bayesian inference gave very similar results. Polymerase chain reaction (PCR)-detected enteroviruses and noroviruses were abundant in post-disinfection effluent (mean concentrations of 2.1 × 10+4–7.2 × 10+5 and 2.7 × 10+4–3.6 × 10+5 gene copies (GC)/L, respectively). Chlorine or UV disinfection produced modest LRs for culture- (0.3–0.9) and PCR-detected enteroviruses (0.3–1.3), as well as noroviruses GI + GII (0.5–0.8). Coliphages and E. coli were more susceptible, with LRs of 0.8–3.0 and 2.5, respectively. Sand-filtered effluent produced significantly higher enteric virus LRs (except cultured enteroviruses). Coliphage and human enteric virus densities gave significantly positive correlations using Kendall's Tau test. Enteric viruses are abundant in wastewater effluent following routine chlorine or UV disinfection processes that target E. coli. Coliphages appear to be good indicators for evaluating wastewater disinfection of enteric viruses.


1988 ◽  
Vol 20 (11-12) ◽  
pp. 25-31 ◽  
Author(s):  
Charles P. Gerba ◽  
Sagar M. Goyal

Densely populated coastal regions of the world generate large quantities of domestic sewage sludge which is often disposed into the marine environment. Present in these wastes are human pathogenic viruses which are capable of surviving for prolonged periods of time in the marine environment and transmitting disease to humans by a number of potential routes of which swimming and consumption of marine foods are the most significant. Hepatitis A virus and Norwalk virus outbreaks associated with shellfish consumption continue to occur in the United States. Three separate epidemiological studies have also shown an association between shellfish consumption and increased risk of hepatitis A infection in consumers. An epidemiological association between non-A and non-B hepatitis has also been demonstrated. A review of the risks of infection, clinical illness and mortality associated with enteroviruses suggests that the presence of these viruses in shellfish and bathing waters presents a significant risk to the consumer.


1985 ◽  
Vol 17 (10) ◽  
pp. 39-41 ◽  
Author(s):  
A. Schnattinger

Ten litres of tapwater were seeded with 200 µl (8×108 HAV particles) of a commercial (Organon Teknika) suspension of hepatitis A virus. Following WALTER and RÜDIGER (1981), the contaminated tapwater was treated with a two-stage technique for concentration of viruses from solutions with low virus titers. The two-stage technique consists of aluminium hydroxideflocculation (200 mg/l Al2(SO4)3. 18 H2O, pH 5,4-5,6) as first stage, the second stage of a lysis of aluminium hydroxidegel with citric acid/sodium citrate-buffer (pH 4,7; 1 ml/l sample), separation of viruses from the lysate by ultracentrifugation and suspension in 1 ml phosphate buffer solution (pH 7,2). A commercial solid phase enzyme-linked immunosorbent assay (ELISA) was used for the detection of HAV. HAV was detecterl in the 10.000:1 concentrates, but not in the seeded 101 samples. Approximately 4×108 of the inoculated 8×108 HAV particles were found in the 1 ml concentrates. The efficiency of detection is about 50%, the virus concentration 5000-fold. Although the percentage loss of HAV in comparison with concentration by means of membrane filtration is similar, the ultracentrifugation method yields a larger sample/concentrate ratio, so that smaller amounts of HAV can be detected more efficiently because of the smaller end-volume.


Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1468
Author(s):  
Saïd Rachida ◽  
Maureen Beatrice Taylor

Hepatitis A virus (HAV) is a waterborne pathogen of public health importance. In South Africa (SA), unique HAV subgenotype IB strains have been detected in surface and wastewater samples, as well as on fresh produce at the point of retail. However, due to the use of molecular-based assays, the infectivity of the detected strains was unknown. Considering the potential shift of HAV endemicity from high to intermediate, which could increase the risk of severe symptomatic disease, this study investigated the identity of HAV strains detected before and after viability treatment of selected wastewater discharge samples. For one year, 118 samples consisting of sewage, treated wastewater discharge and downstream dam water were collected from five wastewater treatment plants (WWTP 1, 2, 3, 4 and 5). Unique HAV IB strains were detected in samples from all five WWTPs, with 11 of these strains carrying amino acid mutations at the immunodominant and neutralisation epitopes. A quasispecies dynamic of HAV has also been detected in sewage samples. The subsequent application of viability PCR revealed that potentially infectious HAV strains were discharged from WWTP 1, 2, 4 and 5 into the dam. Therefore, there is a potential risk of HAV exposure to communities using water sources downstream the WWTPs.


1985 ◽  
Vol 48 (9) ◽  
pp. 815-823 ◽  
Author(s):  
GARY P. RICHARDS

Outbreaks of hepatitis A, Norwalk illness, and nonspecific viral gastroenteritis are associated with consumption of sewage-contaminated shellfish. Over 100 outbreaks have been reported in the United States during the past 50 years. Reported cases of shellfish-associated enteric virus illness are on the increase, whereas bacterial illness from shellfish is on the decline. As yet, there are no procedures for detecting hepatitis A virus, Norwalk virus and numerous other pathogenic viruses in environmental samples, but virus extraction and assay procedures for water and shellfish are available for the more easily cultivated enteric viruses. Current standards rely on bacterial indicators as a means to evaluate the sanitary quality of shellfish and their growing waters, but the adequacy of using bacteria as indicators of possible virus contamination is questionable. The feasibility of employing enteroviruses or rotaviruses as possible viral indiators is discussed. It is proposed that easily cultivated enteroviruses, such as poliovirus, be used as an interim indicator for the possible presence of human pathogenic viruses in seafoods, with the subsequent formulation of guidelines to limit the levels of virus contamination in shellfish.


2000 ◽  
Vol 66 (8) ◽  
pp. 3241-3248 ◽  
Author(s):  
F. Le Guyader ◽  
L. Haugarreau ◽  
L. Miossec ◽  
E. Dubois ◽  
M. Pommepuy

ABSTRACT The main pathogenic enteric viruses able to persist in the environment, such as hepatitis A virus (HAV), Norwalk-like virus (NLV), enterovirus (EV), rotavirus (RV), and astrovirus (AV), were detected by reverse transcription-PCR and hybridization in shellfish during a 3-year study. Oyster samples (n = 108), occasionally containing bacteria, were less frequently contaminated, showing positivity for AV (17%), NLV (23%), EV (19%), and RV (27%), whereas mussel samples, collected in areas routinely impacted by human sewage, were more highly contaminated: AV (50%), HAV (13%), NLV (35%), EV (45%), and RV (52%). Sequences obtained from HAV and NLV amplicons showed a great variety of strains, especially for NLV (strains close to Mexico, Snow Mountain Agent, or Norwalk virus). Viral contamination was mainly observed during winter months, although there were some seasonal differences among the viruses. This first study of virus detection over a fairly long period of time suggests that routine analysis of shellfish by a molecular technique is feasible.


1995 ◽  
Vol 31 (5-6) ◽  
pp. 189-193 ◽  
Author(s):  
Kathleen M. Callahan ◽  
Douglas J. Taylor ◽  
Mark D. Sobsey

The presence and persistence of enteric viruses in sewage contaminated seawater is an important public health concern for bathing, surfing and shellfishing. In an effort to find suitable indicators of enteric viruses in seawater, we compared the survival of two groups of enteric bacteriophages, F-specific coliphages (FRNA phages) and somatic Salmonella bacteriophages (SS phages), to the survival of two human enteric viruses, hepatitis A virus (HAV) and poliovirus type 1 (PV-1), in coastal seawater from three geographic areas (So. California, Hawaii, and North Carolina) at 20°C. Concentrations of all four viruses decreased over 30 days from their initial titers and there was little difference in the survival of a particular virus among the three seawaters. However, the extent of reduction varied among the four viruses. Survival was greater for the SS phages than for any of the other viruses, with an estimated 4 log10 reduction time of about 10 weeks. FRNA phages and PV-1 were inactivated rapidly, with 4 log10 reductions in ~ 1 week. HAV reductions were intermediate between SS phages and FRNA phages, with 4 log10 reductions in about 4 weeks. The observed differences in virus survival suggest that SS phages are more persistent in seawater than other viruses and hence may be good indicators for enteric viruses in seawater.


Author(s):  
Giovanna Fusco ◽  
Aniello Anastasio ◽  
David H. Kingsley ◽  
Maria Grazia Amoroso ◽  
Tiziana Pepe ◽  
...  

To assess the quality of shellfish harvest areas, bivalve mollusk samples from three coastal areas of the Campania region in Southwest Italy were evaluated for viruses over a three-year period (2015–2017). Screening of 289 samples from shellfish farms and other locations by qPCR and RT-qPCR identified hepatitis A virus (HAV; 8.9%), norovirus GI (NoVGI; 10.8%) and GII (NoVGII; 39.7%), rotavirus (RV; 9.0%), astrovirus (AsV; 20.8%), sapovirus (SaV; 18.8%), aichivirus-1 (AiV-1; 5.6%), and adenovirus (AdV, 5.6%). Hepatitis E virus (HEV) was never detected. Sequence analysis identified HAV as genotype IA and AdV as type 41. This study demonstrates the presence of different enteric viruses within bivalve mollusks, highlighting the limitations of the current EU classification system for shellfish growing waters.


2018 ◽  
Vol 23 (7) ◽  
Author(s):  
Maxime Bisseux ◽  
Jonathan Colombet ◽  
Audrey Mirand ◽  
Anne-Marie Roque-Afonso ◽  
Florence Abravanel ◽  
...  

Background Human enteric viruses are resistant in the environment and transmitted via the faecal-oral route. Viral shedding in wastewater gives the opportunity to track emerging pathogens and study the epidemiology of enteric infectious diseases in the community. Aim: The aim of this study was to monitor the circulation of enteric viruses in the population of the Clermont-Ferrand area (France) by analysis of urban wastewaters. Methods: Raw and treated wastewaters were collected between October 2014 and October 2015 and concentrated by a two-step protocol using tangential flow ultrafiltration and polyethylene glycol precipitation. Processed samples were analysed for molecular detection of adenovirus, norovirus, rotavirus, parechovirus, enterovirus (EV), hepatitis A (HAV) and E (HEV) viruses. Results: All wastewater samples (n = 54) contained viruses. On average, six and four virus species were detected in, respectively, raw and treated wastewater samples. EV-positive samples were tested for EV-D68 to assess its circulation in the community. EV-D68 was detected in seven of 27 raw samples. We collected data from clinical cases of EV-D68 (n = 17), HAV (n = 4) and HEV infection (n = 16) and compared wastewater-derived sequences with clinical sequences. We showed the silent circulation of EV-D68 in September 2015, the wide circulation of HAV despite few notifications of acute disease and the presence in wastewater of the major HEV subtypes involved in clinical local cases. Conclusion: The environmental surveillance overcomes the sampling bias intrinsic to the study of infections associated with hospitalisation and allows the detection in real time of viral sequences genetically close to those reported in clinical specimens.


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