Rotavirus In Sewage Research Articles

Environmental Survey of Rotavirus in Sewage Water within Maiduguri Metropolis, Nigeria

Background and study aim: Inadequately treated water supply and unhygienic food consumption are the major sources of contracting feco-orally transmitted viral pathogens, including Rotavirus. This prospective study aimed to assess the presence of Rotavirus in sewage, underground and surface waters collected from different locations within Maiduguri Metropolis, Nigeria. Materials and Methods: A total of 170 sewage and water samples (underground and surface waters) were collected from various sources in high-density and low-density areas of the Maiduguri metropolis. A modified adsorption- elution method for concentration of enteric viruses in sewage and water samples and detection of the presence of rotavirus (if present) was done using Enzyme linked immune-sorbent assay (ELISA). The viruses in water were concentrated by negatively charged membrane filtration, eluted with 0.05M Glycine at pH 11.5, and re-concentrated by centrifugation at 12,000rpm for one hour. Results: Out of the 60 domestic sewage samples collected from high-density areas, 8.3% were positive for rotavirus antigen, whereas none of the samples from low-density areas were positive. In addition, rotavirus antigen was not detected in underground and surface water samples. The temperature range of the 5 rotavirus positive samples was 23-30oC. All the 5 positive samples had alkaline pH that ranged between 8 and 9. Conclusion: This study has demonstrated the persistence of rotavirus in sewage and its possible transmission through contaminated food and poor treated water

Epidemiological Surveillance of Norovirus and Rotavirus in Sewage (2016-2017) in Valencia (Spain).

The aim of the present study was to perform the molecular epidemiology of rotaviruses and noroviruses detected in sewage samples from a large wastewater facility from the city of Valencia, Spain. A total of 46 sewage samples were collected over a one-year period (September 2016 to September 2017). Norovirus and rotavirus were detected and quantified by RT-qPCR, genotyped by semi-nested RT-PCR and further characterized by sequencing and phylogenetic analyses. Noroviruses and rotaviruses were widely distributed in sewage samples (69.6% for norovirus GI, 76.0% norovirus GII, and 71.7% rotaviruses) and viral loads varied from 4.33 to 5.75 log PCRU/L for norovirus GI, 4.69 to 6.95 log PCRU/L for norovirus GII, and 4.08 to 6.92 log PCRU/L for rotavirus. Overall, 87.5% (28/32) of GI noroviruses could not be genotyped, 6.25% (2/32) of the samples contained GI.2 genotype, and another 6.25% (2/32) were positive for GI.4 genotype. The most common genotype of GII noroviruses was GII.2 (40%, 14/35), followed by GII.6 (8.6%, 3/35) and GII.17 (5.7%, 2/35) while the remaining GII strains could not be typed (45.7%, 16/35). Rotavirus VP4 genotype P[8] was the only one found in 19 out of 33 rotavirus-positive samples (57.7%). G2 was the most prevalent rotavirus VP7 genotype (15.2%, 5/33) followed by G3, G9, and G12, with two positive samples for each genotype (6.1%, 2/33). In one sample both G1 and G2 genotypes were detected simultaneously (3%). The results presented here show that the surveillance of noroviruses and rotaviruses in sewage is useful for the study of their transmission in the population and their molecular epidemiology.

Detection of rotavirus in sewage and drinking water by Latex agglutination test

Detection of rotavirus in sewage and drinking water by Latex agglutination test

Detection of enteroviruses, hepatitis A virus and rotaviruses in sewage by means of an immunomagnetic capture reverse transcription-PCR assay

An immunomagnetic capture reverse transcription-PCR (IMC-RT-PCR) assay was evaluated to recover and detect enteric viruses in sewage and to remove PCR inhibitors. The procedure was applied along with a simple sample processing consisting of an initial separation of solids followed by polyethylen glycol precipitation and solvent extraction. This procedure reduced sample volumes by about 65-fold without eliminating RT-PCR inhibitors. Paramagnetic beads coupled to pooled human immunoglobulins were used to simultaneously capture poliovirus 1 (PV-1) and hepatitis A virus (HAV) from seeded sewage concentrates. The IMC was efficient in removing PCR inhibitors and in further reducing sample volumes by approximately 10-fold allowing the analysis of 6-7 ml of sewage sample per RT-PCR reaction. The detection limits of IMC-RT-PCR from seeded concentrates were 0.1-1 PFU for PV-1 and 1 MPNCU for HAV. The described procedure could be applied successfully for the detection of enteroviruses, HAV and rotaviruses in field sewage samples.

Molecular epidemiological survey of rotaviruses in sewage by reverse transcriptase seminested PCR and restriction fragment length polymorphism assay

Rotavirus double-stranded RNA was detected directly in sewage treatment plant samples over a 1-year period by reverse transcription followed by PCR amplification of the VP7 gene and Southern blot hybridization. The presence of naturally occurring rotaviruses was demonstrated in 42% of raw sewage samples and in 67% of treated effluent samples. Amplified viral sequences were analyzed by restriction enzymes. Ten different restriction profiles were characterized, most of which were found in treated effluent samples. A mixture of restriction profiles was observed in 75% of contaminated effluent samples. The profiles were compared with those obtained from human rotavirus isolates involved in infections in children from the same area (six different profiles were detected). Five identical viral sequences were detected in both environmental and clinical samples. Restriction profiles were also compared to profiles from known genomic sequences of human and animal viruses. Both human and animal origins of rotavirus contamination of water seemed likely.

Detection of rotavirus in sewage and creek water: efficiency of the concentration method.

Simian rotavirus SA-11 experimentally seeded, was recovered from raw domestic sewage by a two-step concentration procedure, using filtration through a positively charged microporous filter (Zeta Plus 60 S) followed by ultracentrifugation, effecting an 8,000-fold concentration. By this method, a mean recovery of 81% +/- 7.5 of the SA-11 virus, was achieved.

Genotyping of rotaviruses isolated from sewage.

Rotaviruses from environmental samples have been genotyped by a seminested reverse transcription PCR assay with serotype-specific primers derived from variable regions of gene 9, which produce different characteristic segment sizes for serotypes 1 to 4. The method enabled the detection and identification of type 1, 2, and 3 group A rotaviruses in sewage.

Detection of poliovirus, hepatitis A virus, and rotavirus from sewage and ocean water by triplex reverse transcriptase PCR

A triplex reverse transcriptase PCR (RT-PCR) was developed to simultaneously detect poliovirus, hepatitis A virus (HAV), and rotavirus in sewage and ocean water. Sewage and ocean water samples seeded with the three different viruses were concentrated by ultrafiltration. The unseeded ocean water and sewage samples were concentrated by vortex flow filtration and/or ultrafiltration. Random hexamers and a rotavirus downstream primer were used to initiate reverse transcription. Three different sets of primers specific for poliovirus, HAV, and rotavirus cDNAs were mixed in the PCR mixture to amplify the target DNA. Three distinct amplified DNA products representing poliovirus, HAV, and rotavirus were identified by gel electrophoresis as 394-, 192-, and 278-bp sequences, respectively. Dot blot and Southern analyses were used to confirm the amplified products for each virus present in the environmental samples. Except for poliovirus, the sensitivity of triplex RT-PCR for the detection of rotavirus and HAV was found to be similar to that of monoplex RT-PCR, which uses only one set of primers to amplify a single type of virus. The triplex RT-PCR has greater advantages over monoplex RT-PCR for virus detection, namely, the rapid turnaround time and cost effectiveness.

Detection of human rotavirus in sewage through two concentration procedures

Abstract Two different methodologies for the detection of human rotavirus in water were compared. Simian rotavirus SA11 was used to develop these procedures in the laboratory with distilled water, tap water and raw sewage. The adsorption to and elution from glass powder method was used to concentrate seeded rotavirus from 20-1. samples of water and yielded satisfactory recoveries of viruses from distilled and tap water (54 and 46% respectively) but poor recovery (5%) from raw sewage. In the second concentration methodology, rotaviruses were concentrated from 600-ml samples by ammonium sulfate flocculation after the addition to samples of beef extract up to a 10% final concentration. Rotaviruses were recovered from raw sewage without any substantial loss compared to distilled and tap water (67% in raw sewage, and 70 and 73% in distilled and tap water). The ammonium sulfate method consistently yielded greater viral recoveries than the adsorption-elution method, although the latter enables the processing of larger volume samples. Both methodologies were applied to detect human rotavirus in raw sewage at two Barcelona wastewater effluents. Human rotavirus in sewage isolates was quantitated by immunofluorescence in the MA-104 cell line. Indigenous rotavirus was detected in 100% of sewage samples using both different concentration procedures; rotaviruses were also found in 69% of nonconcentrated samples. The level of rotaviruses found in Barcelona sewage was very high (up to 14,000 fluorescent foci 1 −1 ), being of the same magnitude as the level of enteroviruses, although no correlation was found with the levels of human enteric viruses.

Seasonal occurrence of rotavirus in sewage

A seasonal distribution was observed for rotaviruses in sewage by using indirect immunofluorescence. Levels were low from May through September and generally higher during winter and spring. In contrast, no seasonal pattern was observed for total enteroviruses. Limitations of the indirect immunofluorescence assay and enzyme immunoassay for environmental samples are discussed.

Detection of Rotavirus in Sewage

FOR DETECTION OF ROTAVIRUS, DOMESTIC SEWAGE WAS CONCENTRATED BY TWO DIFFERENT METHODS: (i) adsorption to and elution from positively charged Seitz filters, followed by ultracentrifugation, and (ii) chemical precipitation. The concentrated fluids were tested by an enzyme-linked immunosorbent assay and electron microscopy. In 6 of 24 (25%) samples, rotavirus was detectable after the combined filtration and ultracentrifugation technique with both an enzyme-linked immunosorbent assay and electron microscopy. No positive results were obtained after chemical precipitation.