Effect of the Shellfish Proteinase K Digestion Method on Norovirus Capsid Integrity

Norovirus outbreaks are associated with the consumption of contaminated shellfish, and so efficient methods to recover and detect infectious norovirus in shellfish are important. The Proteinase K digestion method used to recover norovirus from shellfish, as described in the ISO 15216, would be a good candidate but its impact on the virus capsid integrity and thus infectivity was never examined. The aim of this study was to assess the impact of the Proteinase K digestion method, and of the heat treatment component of the method alone, on norovirus (genogroups I and II) and MS2 bacteriophage capsid integrity. A slightly modified version of the ISO method was used. RT-qPCR was used for virus detection following digestion of accessible viral RNA using RNases. MS2 phage infectivity was measured using a plaque assay. The effect of shellfish digestive glands (DG) on recovery was evaluated. In the presence of shellfish DG, a reduction in MS2 phage infectivity of about 1 log₁₀ was observed after the Proteinase K digestion method and after heat treatment component alone. For norovirus GII and MS2 phage, there was no significant loss of genome following the Proteinase K digestion method but there was a significant 0.24 log₁₀ loss of norovirus GI. In the absence of shellfish DG, the reduction in MS2 phage infectivity was about 2 log₁₀, with the addition of RNases resulting in a significant loss of genome for all tested viruses following complete Proteinase K digestion method and the heat treatment alone. While some protective effect from the shellfish DG on viruses was observed, the impact on capsid integrity and infectivity suggests that this method, while suitable for norovirus genome detection, may not completely preserve virus infectivity.     

Road effects on abundance and fitness of Galápagos lava lizards (Microlophus albemarlensis)

Vehicle collisions with wildlife have the potential to negatively impact species everywhere roads divide natural habitats; this is especially true in areas where roads are a recent addition to the landscape (e.g., in the last decade) and/or where expansion of a road network is underway. This study addresses the impact of traffic fatalities on lava lizards, Microlophus albemarlensis, on the island of Santa Cruz, Galápagos. The main road bisecting the island north to south was added to the island beginning in 1974 but not fully paved until 2000. We assessed impacts occurring due to traffic and vehicle speeds on the island. This study adds impacts to lizard populations to those conducted on birds on Santa Cruz, providing a means to create multi-species recommendations. To quantify impacts to lizard populations, we assessed lizards on randomly chosen transects perpendicular to the road. We used Poisson regression to analyze those data and found a 30% increase in lizard abundance per 100 m distance from the road across vegetation zones. We compared incidence of prior tail loss of road-killed lizards and live animals to comment on additional energetic costs of occupying near-road territories. Tail condition could not be assessed in some road-killed lizards; among those that we could assess, 29% showed evidence of prior tail loss. That rate declined to apparent baseline conditions of only 1% beyond 200 m from the road. By traveling the length of the 40-km road, we identified hot spots for fatalities of both lizards and birds to provide necessary information for developing management and remediation strategies. We offer guidance on Best Management Practices, such as testing overpasses, fencing and underpasses, which might reduce vehicle impacts to lizards from drivers on the current road. This information is essential because of a planned increase in the Santa Cruz road network in areas where lava lizards are most abundant.