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  1. This study suggests a new method for determining the viability of Ascaris spp. ova, based on in-vitro early-to-late stage development of ova. This method includes stages prior to larval development, providing an estimation of potential viability.After application of biosolids onto soil and exposure to 7°C, 22°C, or 37°C for 45 days, ova were microscopically distinguished as viable or non-viable according to progression through development categories. Results were compared to viability estimates from current methods that distinguish viable ova as motile larva. Results suggest conventional techniques underestimate viability, whereas the new method provides a more conservative approach. 
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  2. The present study investigated wastewater treatment for the removal of 11 different virus types (pepper mild mottle virus; Aichi virus; genogroup I, II, and IV noroviruses; enterovirus;sapovirus; group-A rotavirus; adenovirus; and JC and BK polyomaviruses) by two wastewater treatment facilities utilizing advanced Bardenpho technology and compared the results with conventional treatment processes. To our knowledge, this is the first study comparing full-scale treatment processes that all received sewage influent from the same region. The incidence of viruses in wastewater was assessed with respect to absolute abundance, occurrence, and reduction in monthly samples collected throughout a 12 month period in southern Arizona. Samples were concentrated via an electronegative filter method and quantified using TaqMan-based quantitative polymerase chain reaction (qPCR). Results suggest that Plant D, utilizing an advanced Bardenpho process as secondary treatment, effectively reduced pathogenic viruses better than facilities using conventional processes. However, the absence of cell-culture assays did not allow an accurate assessment of infective viruses. On the basis of these data, the Aichi virus is suggested as a conservative viral marker for adequate wastewater treatment, as it most often showed the best correlation coefficients to viral pathogens, was always detected at higher concentrations, and may overestimate the potential virus risk. 
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  3. We investigated the occurrence and attenuation of several human enteric viruses (i.e., norovirus, adenovirus,Aichi virus 1, polyomaviruses, and enterovirus) as well as a plant virus, pepper mild mottle virus (PMMoV), at two surface flow wetlands in Arizona. The retention time in one of the wetlands was seven days, whereas in the otherwetland it could not be defined.Water sampleswere collected at the inlet and outlet fromthewetlands over nine months, and concentration of viral genomes was determined by quantitative polymerase chain reaction (qPCR). Of the human enteric viruses tested, adenovirus and Aichi virus 1 were found in the greatest prevalence in treated wastewater (i.e., inlet of thewetlands). Reduction efficiencies of enteric viruses by thewetlands ranged from1 to 3 log10. Polyomaviruseswere generally removed to belowdetection limit, indicating at least 2 to 4 log10 removal. PMMoV was detected in a greater concentration in the inlet of both wetlands for all the viruses tested (104 to 107 genome copies/L), but exhibited little or no removal (1 log10 or less). To determine the factors associated with virus genome attenuation (as determined by qPCR), the persistence of PMMoV and poliovirus type 1 (an enterovirus) was studied in autoclaved and natural wetland water, and deionized water incubated under three different temperatures for 21 days. A combination of elevated water temperature and biological activities reduced poliovirus by 1 to 4 log10, while PMMoV was not significantly reduced during this time period.Overall, PMMoV showed much greater persistence than human viruses in the wetland treatment. 
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