Limits...
The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge.

Prado T, Guilayn Wde C, Gaspar AM, Miagostovich MP - Mem. Inst. Oswaldo Cruz (2013)

Bottom Line: Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion.Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control.This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Virologia Comparada e Ambiental, Instituto Oswaldo Cruz-Fiocruz, Rio de Janeiro, RJ, Brasil. tprado@ioc.fiocruz.br

ABSTRACT
The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods) used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR) was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%), RVA, NoV GII (45%) and HAV (18%), indicating the viruses' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

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recovery efficiency (%) from each method performed in trip licate used for detecting enteric viruses. AdV: adenoviruses; HAV: hepatitis A virus; Method 1: ultracentrifugation; Method 2: beef ex tract; NoV GII: norovirus genogroup II; RVA: rotavirus species A.
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f02: recovery efficiency (%) from each method performed in trip licate used for detecting enteric viruses. AdV: adenoviruses; HAV: hepatitis A virus; Method 1: ultracentrifugation; Method 2: beef ex tract; NoV GII: norovirus genogroup II; RVA: rotavirus species A.

Mentions: PP7 was recovered from all of the analysed samples and the mean VLs detected by Methods 1 and 2 were as follows: 5.2 x 106 GC g-1 ± SD 1.1 x 106 and 2.8 x 106 GC g-1 ± SD 2.3 x 106, respectively. When the nucleic acids were diluted 1:100 in Method 2, a higher mean recovery rate was obtained for PP7: 1.3 x 107 ± SD 1.1 x 107 GC g-1 (Fig. 2).


The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge.

Prado T, Guilayn Wde C, Gaspar AM, Miagostovich MP - Mem. Inst. Oswaldo Cruz (2013)

recovery efficiency (%) from each method performed in trip licate used for detecting enteric viruses. AdV: adenoviruses; HAV: hepatitis A virus; Method 1: ultracentrifugation; Method 2: beef ex tract; NoV GII: norovirus genogroup II; RVA: rotavirus species A.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3974313&req=5

f02: recovery efficiency (%) from each method performed in trip licate used for detecting enteric viruses. AdV: adenoviruses; HAV: hepatitis A virus; Method 1: ultracentrifugation; Method 2: beef ex tract; NoV GII: norovirus genogroup II; RVA: rotavirus species A.
Mentions: PP7 was recovered from all of the analysed samples and the mean VLs detected by Methods 1 and 2 were as follows: 5.2 x 106 GC g-1 ± SD 1.1 x 106 and 2.8 x 106 GC g-1 ± SD 2.3 x 106, respectively. When the nucleic acids were diluted 1:100 in Method 2, a higher mean recovery rate was obtained for PP7: 1.3 x 107 ± SD 1.1 x 107 GC g-1 (Fig. 2).

Bottom Line: Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion.Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control.This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

View Article: PubMed Central - PubMed

Affiliation: Laboratório de Virologia Comparada e Ambiental, Instituto Oswaldo Cruz-Fiocruz, Rio de Janeiro, RJ, Brasil. tprado@ioc.fiocruz.br

ABSTRACT
The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods) used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR) was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%), RVA, NoV GII (45%) and HAV (18%), indicating the viruses' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

Show MeSH
Related in: MedlinePlus