ABSTRACT
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 x 10(4) to 1.2 x 10(6) and from 50 to 1.3 x 10(4) gc/ml in sludge and wastewater, respectively. HAV quantification in sludge ranged from 3.1 x 10(2) to 5.4 x 10(2) gc/ml. In conclusion, it was possible to correlate presence and viability of enteric viruses in the environmental samples analyzed.
Subject(s)
Polymerase Chain Reaction/methods , Sewage/virology , Viruses/isolation & purification , Waste Disposal, Fluid/methods , Water Microbiology , Animals , Brazil , Cell Line , DNA, Viral/isolation & purification , Humans , RNA, Viral/isolation & purification , Seasons , Time FactorsABSTRACT
The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20% of the AdV, 90% of the RV and 100% of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8% of the AdV and 90% of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.
Subject(s)
Adenoviruses, Human/isolation & purification , RNA Viruses/isolation & purification , Sewage/virology , Water Microbiology , DNA, Viral/isolation & purification , Hepatitis A virus/isolation & purification , Poliovirus/isolation & purification , RNA, Viral/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction , Rotavirus/isolation & purificationABSTRACT
The aim of this study was to compare two nucleic acid extraction methods for the recovery of enteric viruses from activated sludge. Test samples were inoculated with human adenovirus (AdV), hepatitis A virus (HAV), poliovirus (PV) and rotavirus (RV) and were then processed by an adsorption-elution-precipitation method. Two extraction methods were used: an organic solvent-based method and a silica method. The organic-based method was able to recoup 20 percent of the AdV, 90 percent of the RV and 100 percent of both the PV and HAV from seeded samples. The silica method was able to recoup 1.8 percent of the AdV and 90 percent of the RV. These results indicate that the organic-based method is more suitable for detecting viruses in sewage sludge.
