[Show abstract][Hide abstract] ABSTRACT: Rotavirus virus-like particles (VLPs) and MS2 bacteriophages were bioaccumulated in bivalve mollusks to evaluate viral persistence
in shellfish during depuration and relaying under natural conditions. Using this nonpathogenic surrogate virus, we were able
to demonstrate that about 1 log10 of VLPs was depurated after 1 week in warm seawater (22°C). Phage MS2 was depurated more rapidly (about 2 log10 in 1 week) than were VLPs, as determined using a single-compartment model and linear regression analysis. After being relayed
in the estuary under the influence of the tides, VLPs were detected in oysters for up to 82 days following seeding with high
levels of VLPs (concentration range between 1010 and 109 particles per g of pancreatic tissue) and for 37 days for lower contamination levels (105 particles per g of pancreatic tissue). These data suggest that viral particles may persist in shellfish tissues for several
Preview · Article · Nov 2005 · Applied and Environmental Microbiology
[Show abstract][Hide abstract] ABSTRACT: The potential of rotavirus 2/6-virus-like-particles (VLP2/6) for use as tracers in the marine environment was investigated. The stability of bovine rotavirus (strain RF) and VLP2/6 in natural seawater at 25 degrees C for six days was studied. ELISA and western blot methods were used to quantify the particles. The rates of decline of rotavirus particles and VLP2/6 were similar (approximately 0.5log(10) per day). Western blot analysis showed that the integrity of capsid proteins VP2 and VP6 was conserved during the incubation time. These results demonstrate that VLP2/6 particles have the same stability in seawater as rotavirus particles. Thus, VLP2/6 can be used as a tracer, which should be of particular value for studying the fate of rotavirus particles in the marine environment.
Full-text · Article · Oct 2004 · Research in Microbiology
[Show abstract][Hide abstract] ABSTRACT: Virus-like particles (VLPs) with the full-length VP2 and VP6 rotavirus capsid proteins, produced in the baculovirus expression system, have been evaluated as surrogates of human rotavirus in different environmental scenarios. Green fluorescent protein-labeled VLPs (GFP-VLPs) and particles enclosing a heterologous RNA (pseudoviruses), whose stability may be monitored by flow cytometry and antigen capture reverse transcription-PCR, respectively, were used. After 1 month in seawater at 20 degrees C, no significant differences were observed between the behaviors of GFP-VLPs and of infectious rotavirus, whereas pseudovirus particles showed a higher decay rate. In the presence of 1 mg of free chlorine (FC)/liter both tracers persisted longer in freshwater at 20 degrees C than infectious viruses, whereas in the presence of 0.2 mg of FC/liter no differences were observed between tracers and infectious rotavirus at short contact times. However, from 30 min of contact with FC onward, the decay of infectious rotavirus was higher than that of recombinant particles. The predicted Ct value for a 90% reduction of GFP-VLPs or pseudoviruses induces a 99.99% inactivation of infectious rotavirus. Both tracers were more resistant to UV light irradiation than infectious rotavirus in fresh and marine water. The effect of UV exposure was more pronounced on pseudovirus than in GFP-VLPs. In all types of water, the UV dose to induce a 90% reduction of pseudovirus ensures a 99.99% inactivation of infectious rotavirus. Recombinant virus surrogates open new possibilities for the systematic validation of virus removal practices in actual field situations where pathogenic agents cannot be introduced.
Full-text · Article · Aug 2004 · Applied and Environmental Microbiology