SARS-CoV-2 infection augments species- and age-specific predispositions in cotton rats.

Heterogeneity of COVID-19 manifestations in human population is vast, for reasons unknown. Cotton rats are a clinically relevant small animal model of human respiratory viral infections. Here, we demonstrate for the first time that SARS-CoV-2 infection in cotton rats affects multiple organs and systems, targeting species- and age-specific biological processes. Infection of S. fulviventer, which developed a neutralizing antibody response and were more susceptible to SARS-CoV-2 replication in the upper respiratory tract, was accompanied by hyperplasia of lacrimal drainage-associated lymphoid tissue (LDALT), a first known report of mucosa-associated lymphoid tissue activation at the portal of SARS-CoV-2 entry. Although less permissive to viral replication, S. hispidus showed hyperplasia of bone marrow in the facial bones and increased pulmonary thrombosis in aged males. Augmentation of these features by SARS-CoV-2 infection suggests a virus-induced breach in regulatory mechanisms which could be devastating for people of all ages with underlying conditions and in particular for elderly with a multitude of ongoing disorders.

Observed Reductions in the Infectivity of Bioaerosols Containing Bovine Coronavirus Under Repetitively Pulsed RF Exposure.

OBJECTIVE: The purpose of the present study is to investigate the inactivation of bioaerosols containing Bovine Coronavirus, BCoV, under repetitively pulsed radio frequency (RF) electromagnetic exposure. METHODS: These experiments were performed in a waveguide containing a flowing aerosol stream and were limited to a single RF waveform: ∼2 µs square envelope, 5.6 GHz, 4.8 kHz repetition rate. Aerosol streams were exposed to RF electric field amplitudes in the range of 41.9 +/-6.2 kV/m. Under laminar flow conditions, 75% of the total collected aerosol stream spends 0.85 seconds or less in the RF exposure region. RESULTS: Application of the RF waveform changed mean survival rate of the aerosolized BCoV by -0.58 decades (roughly a 74% reduction) and impacted the variance and standard deviation of the experimental results, with the RF exposure data showing an 800% increase in variance and 196% increase in standard deviation over the control results. Experimental results were compared to those from an analytic electromagnetic-heating inactivation model. CONCLUSION: The comparison indicated the feasibility that the observed reduction in BCoV survival rate might be due to a combination of thermal effects and non-thermal electric field effects. SIGNIFICANCE: Developing better insight into the mechanisms of inactivation is important for understanding the potential limits of efficacy for this method. Additionally, these results contribute an important baseline for the impact of electromagnetic fields on aerosolized pathogens.

A conserved domain in the coronavirus membrane protein tail is important for virus assembly.

Coronavirus membrane (M) proteins play key roles in virus assembly, through M-M, M-spike (S), and M-nucleocapsid (N) protein interactions. The M carboxy-terminal endodomain contains a conserved domain (CD) following the third transmembrane (TM) domain. The importance of the CD (SWWSFNPETNNL) in mouse hepatitis virus was investigated with a panel of mutant proteins, using genetic analysis and transient-expression assays. A charge reversal for negatively charged E(121) was not tolerated. Lysine (K) and arginine (R) substitutions were replaced in recovered viruses by neutrally charged glutamine (Q) and leucine (L), respectively, after only one passage. E121Q and E121L M proteins were capable of forming virus-like particles (VLPs) when coexpressed with E, whereas E121R and E121K proteins were not. Alanine substitutions for the first four or the last four residues resulted in viruses with significantly crippled phenotypes and proteins that failed to assemble VLPs or to be rescued into the envelope. All recovered viruses with alanine substitutions in place of SWWS residues had second-site, partially compensating, changes in the first TM of M. Alanine substitution for proline had little impact on the virus. N protein coexpression with some M mutants increased VLP production. The results overall suggest that the CD is important for formation of the viral envelope by helping mediate fundamental M-M interactions and that the presence of the N protein may help stabilize M complexes during virus assembly.