Search

Human noroviruses (hNoV) are the primary cause of foodborne disease in the USA. Most studies on inactivation kinetics of hNoV and its surrogates are performed in monoculture, while the microbial ecosystem effect on virus inactivation remains limited. This study investigated the persistence of hNoV surrogates, murine norovirus (MNV) and Tulane virus (TuV), along with Aichi virus (AiV) under thermal and chemical inactivation in association with Gram-negative (Enterobacter cloacae) bacteria. Thermal inactivation of viruses in co-culture with E. cloacae revealed no protective effects of bacteria. At 56 °C, AiV with and without bacteria was completely inactivated by 10 min with decimal reduction values (D-values) of 41 and 43 s, respectively. Similar results were also observed for TuV. Conversely, MNV with bacteria was completely inactivated by 10 min while MNV alone remained stable up to 30 min at 56 °C. Both MNV and TuV were slightly more stable than AiV at 63 °C with TuV detection up to 2 min without bacteria. For chemical inactivation on stainless steel surfaces, viruses alone and in association with bacteria were treated with 1000 ppm sodium hypochlorite. Virus association with bacteria had no significant effect (p > 0.05) on virus resistance to bleach inactivation compared to virus alone. Specifically, exposure to 1000 ppm bleach for 5 min resulted in an average of 3.86, 2.14, and 0.94 log 10 PFU/ml reductions for TuV, MNV, and AiV without bacteria, respectively. Reductions in TuV, MNV, and AiV were 3.50, 1.88, and 0.61 log10 PFU/ml when associated with E. cloacae, respectively.

Ultrafiltration (UF) membranes are increasingly being used for wastewater reclamation treatment for their high removal of pathogens and suspended solids. However, breakage of UF membrane fibers could allow leakage of pathogens into the permeate and create health risks in the use of reclaimed water. Here, we assessed the log 10 reduction value (LRV) of human enteric viruses and microbial indicators of new and aged UF modules in a pilot-scale UF process to evaluate the influence of fiber breakage. Norovirus genotypes I and II, Aichi virus, and Escherichia coli were not detected in any permeate samples of intact UF modules, but were detected in samples of damaged UF modules. LRVs of all microorganisms assayed decreased as fiber breakage of new UF modules increased, with maximum decreases of > 3.3 log 10 . Fiber breakage in the aged UF modules did not decrease LRVs of somatic coliphages and MS2, but breakage in the new UF modules did decrease them. Intact new UF modules gave higher LRVs than intact aged UF modules. When the LRV of intact UF module was assumed to be 1 or 2 log 10 , increasing fiber breakage did not significantly decrease the predicted LRV, but when it was ≥ 3 log 10 , it did decrease LRV, in good agreement with measured LRVs in the degraded UF modules. These results suggest that the LRV of intact UF modules affects the decrease in LRV and confirm the leakage of human enteric viruses following fiber breakage in UF modules of different ages in the UF process of wastewater reclamation.

Phosphatidylinositol 4-kinase III beta (PI4KIIIβ) is an essential enzyme in mediating membrane transport, and plays key roles in facilitating viral infection. Many pathogenic positive-sense single-stranded RNA viruses activate PI4KIIIβ to generate phosphatidylinositol 4-phosphate (PI4P)-enriched organelles for viral replication. The molecular basis for PI4KIIIβ activation during viral infection has remained largely unclear. We describe the biochemical reconstitution and characterization of the complex of PI4KIIIβ with the Golgi protein Acyl-coenzyme A binding domain containing protein 3 (ACBD3) and Aichi virus 3A protein on membranes. We find that 3A directly activates PI4KIIIβ, and this activation is sensitized by ACBD3. The interfaces between PI4KIIIβ-ACBD3 and ACBD3-3A were mapped with hydrogen-deuterium exchange mass spectrometry (HDX-MS). Determination of the crystal structure of the ACBD3 GOLD domain revealed a unique N terminus that mediates the interaction with 3A. Rationally designed complex-disrupting mutations in both ACBD3 and PI4KIIIβ completely abrogated the sensitization of 3A activation by ACBD3., (Copyright © 2016 Elsevier Ltd. All rights reserved.)