Norwalk-Like Virus Sequences in Mineral Waters: One-Year Monitoring of Three Brands

Cantonal Food Laboratory of Solothurn, CH-4500 Solothurn, Switzerland.
Applied and Environmental Microbiology (Impact Factor: 3.67). 05/2002; 68(4):1925-31. DOI: 10.1128/AEM.68.4.1925-1931.2002
Source: PubMed


In a recent study, RNA with nucleotide sequeces specific for "Norwalk-like viruses" (NLV) was detected in 11 different brands of European mineral waters. To clarify this finding, a 1-year monitoring study was conducted. Samples of three European brands of mineral water without gas were monitored weekly by reverse transcriptase PCR using generic and genogroup-specific oligonucleotides. Additional analyses were performed to investigate a possible correlation between NLV sequence contamination and mineral water lot numbers, the long-term stability (persistence) of NLV sequences in mineral water, and the level of contamination. NLV sequences were detected in 53 of 159 samples analyzed (33%) and belonged entirely to genogroup II. Although all NLV strains identified were closely related, three mineral water brand-specific clusters could be identified for both primer systems by sequencing. Analyses of second samples from lots previously shown to be positive for NLV sequences gave corresponding results in 45 of 53 cases (85%) (within a six-pack). NLV persistence was tested by analyzing 10 positive samples after 6 and 12 months of storage in darkness at room temperature. After 6 months, all samples remained positive; after 12 months, 9 of 10 samples were still positive for NLV sequences. No NLV sequences could be detected by analysis of 0.1-liter aliquots of 53 samples shown to be positive by testing of 1-liter volumes. Based on this fact and a test sensitivity of approximately 10 viral units, levels of contamination in positive mineral water samples were estimated to be in the range of 10 to 100 genomic equivalents per liter.

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Available from: Christian Beuret, Feb 07, 2014
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    • "Even tertiary-treated reclaimed wastewater has been shown to contain viable Cryptosporidium oocysts (Quintero-Betancourt et al., 2003). Similarly, some reports indicated that even mineral water may be contaminated with Norwalk-like viruses (Beuret et al., 2002), thereby raising questions on the technologies being used at water treatment plants for detection and disinfection. "
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    ABSTRACT: Thousands of tons of animal and human wastes are produced every day across the globe. Their continuous use as fertilizers or soil amendments on agricultural lands has raised severe health and environmental concerns. Incorporating bio-wastes into the soil is one of the best management strategies to control the run-off of nutrients from the soil surface and, in turn, protect freshwater. However, exposing bio-waste to sunlight in the field is considered to be one of the most cost-effective methods to control pathogens. Considering the differences among different bio-waste management strategies, there is a need for a review of the current knowledge on the practice of spreading bio-wastes onto the soil surface. Specifically, this information would help us to better understand the fate of pathogens upon their exposure to the open environment, how the presence of bio-waste on the soil surface can threaten humans and the environment, and the costs and benefits of surface-applied bio-wastes. Current review of the literature revealed a lack of understanding of the factors responsible for killing pathogens on the soil surface. More than 150 pathogens (including different viruses, bacteria, protozoan and helminth) have been reported to be present in different bio-wastes, but the majority of studies have focused on a few common pathogens. Similarly, over the last decade, each year at least 1 new pathogen is being reported which can threat public health but there is a paucity of knowledge concerning the fate of pathogens under field conditions. Similarly, the techniques used for the detection of pathogens were found to be variable and inconclusive, making it difficult to compare the results of different studies. Therefore, given that the tools for the evaluation of pathogens have serious limitations and the survival characteristics of many (old and emerging) pathogens are yet to be discovered, the spreading of bio-wastes (treated or untreated) onto the soil surface (i.e., unincorporated) may not only further increase the threat for human health but also further aggravate the environment.
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    • "The percentage of annual detection ranged from 19.5% to 32.7%, except in 2006 when this percentage reached 47.4% (659/1389). NoV were detected using the region B (RNA-dependent RNA-polymerase) RT-PCR protocol previously described [43] and genotyped as NoV GII.4 by the partial nucleotide sequencing of region D (VP1) [44]. A total of 147 NoV GII.4 samples previously characterized were selected from 12 states of the three most highly populated regions of Brazil (Northeast, South, and Southeast), representing approximately 85% of the countrýs population (190.732.694 "
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    PLoS ONE 03/2014; 9(3):e92988. DOI:10.1371/journal.pone.0092988 · 3.23 Impact Factor
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    • "Reverse transcription was carried out using random primers (Invitrogen ® , Carlsbad, CA, USA). For NoV detection, generic primers (Mon431/Mon432 and Mon433/Mon434) that target the region B of the ORF1 (RdRp gene) [7], were used according to the conditions described by Beuret et al. [19]. The molecular characterization was performed by using degenerated primers CapA/CapB1/CapB2 and CapC/CapD1/CapD3 that target the VP1 gene (region D) as described by Vinjé et al. [9]; and the primers JV12Y, JV12i, Ni-R and G1 that target the RdRp gene as described previously by Boxman et al. [10]. "
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