Single-pass wind tunnel testing for recirculating virus aerosol control technologies

A number of recirculating flow aerosol control technologies have been commercialized to mitigate aerosol-transmitted virus infections. Many of these technologies incorporate filters for particle collection and some may also incorporate technologies for virus inactivation. Given the wide variety of commercially available aerosol control technologies to consumers, it is extremely important to develop standardized methods to characterize their performance in bioaerosol removal and inactivation, such that technologies can be compared on an “equivalent-test” basis. However, no standard procedures have been established to evaluate the effectiveness of bioaerosol removal and inactivation in recirculating aerosol control technologies. We propose the use of a single-pass tunnel to assess the performance of bioaerosol control technologies, as single-pass wind tunnels can be sealed with well-controlled velocity and particle concentration profiles. Here, we specifically describe the construction of a single-pass wind tunnel and apply it to three recirculating aerosol control technologies, incorporating UV-C sources, filters, and electrostatic precipitators, respectively. We utilize a porcine respiratory coronavirus (PRCV) challenge aerosol, generated via pneumatic nebulization of a high titer (∼107 TCID50 mL−1) viral suspension. Following guidelines similar to those used in the ANSI/ASHRAE Standard 52-2 test procedure for HVAC filters, in single-pass wind tunnel tests, velocity uniformity and particle uniformity are first monitored across the cross-section of the tunnel. The size distribution of viable particles is additionally determined in advance of tests by the collection of particles in the wind tunnel using a cascade impactor, with both RT-qPCR and titration used to quantify viruses collected on each impaction stage. We show that the viable particle size distribution follows the volumetric size distribution of the nebulized virus-laden suspension, and that this distribution can be tuned to be similar in shape to the observed distribution of aerosol from human respiratory activities. Following tunnel and virus aerosol characterization, for each tested technology, using triplicate tests, the single-pass log reduction based on RT-qPCR and viable virus titration is determined by simultaneously collecting virus aerosol particles upstream and downstream of the control technology. The tested technologies in this study have titration-based single-pass log reductions in the 1.5–4.0 range. Overall, design and testing suggest that the single-pass wind tunnel approach is a tractable method to examine the efficacy of aerosol control technologies in removing and inactivating viruses in aerosols, and suggest that such technologies should be described by their single-pass log reduction and operating flow rate, with the test virus size distribution reported alongside test results. In addition, we examine the limits of detection in single-pass wind tunnel tests in comparison to chamber tests, and in doing so find that for most control technologies, the wind tunnel test will yield higher concentrations downstream or during sampling, and hence clearer results for the log reduction.

Greater than 3-Log Reduction in Viable Coronavirus Aerosol Concentration in Ducted Ultraviolet-C (UV-C) Systems.

Control technologies to inactivate airborne viruses effectively are needed during the ongoing SARS-CoV-2 pandemic, and to guard against airborne transmitted diseases. We demonstrate that sealed UV-C flow reactors operating with fluences near 253 ± 1 nm of 13.9-49.6 mJ cm-2 efficiently inactivate coronaviruses in an aerosol. For measurements, porcine respiratory coronavirus (PRCV) was nebulized in a custom-built, 3.86 m wind tunnel housed in a biosafety level class II facility. The single pass log10 reduction of active coronavirus was in excess of 2.2 at a flow rate of 2439 L min-1 (13.9 mJ cm-2) and in excess of 3.7 (99.98% removal efficiency) at 684 L min-1 (49.6 mJ cm-2). Because virus titers resulting from sampling downstream of the UV-C reactor were below the limit of detection, the true log reduction is likely even higher than measured. Comparison of virus titration results to reverse transcriptase quantitative PCR and measurement of fluorescein concentrations (doped into the nebulized aerosol) reveals that the reduction in viable PRCV is primarily due to UV-C based inactivation, as opposed to physical collection of virus. The results confirm that UV-C flow reactors can efficiently inactivate coronaviruses through incorporation into HVAC ducts or recirculating air purifiers.

Wind tunnel-based testing of a photoelectrochemical oxidative filter-based air purification unit in coronavirus and influenza aerosol removal and inactivation.

Recirculating air purification technologies are employed as potential means of reducing exposure to aerosol particles and airborne viruses. Toward improved testing of recirculating air purification units, we developed and applied a medium-scale single-pass wind tunnel test to examine the size-dependent collection of particles and the collection and inactivation of viable bovine coronavirus (BCoV, a betacoronavirus), porcine respiratory coronavirus (PRCV, an alphacoronavirus), and influenza A virus (IAV), by a commercial air purification unit. The tested unit, the Molekule Air Mini, incorporates a MERV 16 filter as well as a photoelectrochemical oxidating layer. It was found to have a collection efficiency above 95.8% for all tested particle diameters and flow rates, with collection efficiencies above 99% for supermicrometer particles with the minimum collection efficiency for particles smaller than 100 nm. For all three tested viruses, the physical tracer-based log reduction was near 2.0 (99% removal). Conversely, the viable virus log reductions were found to be near 4.0 for IAV, 3.0 for BCoV, and 2.5 for PRCV, suggesting additional inactivation in a virus family- and genus-specific manner. In total, this work describes a suite of test methods which can be used to rigorously evaluate the efficacy of recirculating air purification technologies.

Immune responses to porcine epidemic diarrhea virus (PEDV) in swine and protection against subsequent infection.

Understanding the immune responses against Porcine epidemic diarrhea virus (PEDV) is important to prevent infection and to design control strategies. We evaluated both systemic and mucosal immune responses to PEDV in pigs and assessed if prior exposure to virus protects against re-infection. Three-week-old pigs were infected with PEDV and immune response in blood, intestine, and mesenteric lymph node (MLN) was evaluated. At 30 dpi, virus exposed pigs were challenged with a field isolate of PEDV and immune response at 5 d post challenge was evaluated. We found that PEDV RNA persists in the intestine even after fecal shedding of the virus was stopped at 28 dpi and pigs previously exposed to PEDV are protected from virus shedding after re-infection. PEDV infection induced both humoral and cell mediated immune response with an increase in PEDV specific IgA and IgG antibodies in intestine and serum. Flow cytometry analysis showed a significantly higher frequency of B cells and lower frequency of T cells at 4 dpi. The frequency of CD4/CD8 double positive (DP) memory T cells was significantly increased in the MLN of challenged animals. These studies may provide further insights into understanding the mucosal immune response to PEDV and its role in protection against disease.