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      Aerosol Generation during Spirometry

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          Abstract

          The coronavirus disease (COVID-19) pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has severely restricted pulmonary diagnostic testing because of the concern of droplet and aerosol generation by procedures conducted in small test rooms. SARS-CoV-2 infection is characterized by viral shedding from the upper and lower respiratory tracts; in addition, SARS-CoV-2 RNA has been detected in sampled air throughout a hospital, which leads to this concern (1–4). Pulmonary function laboratories are justifiably concerned because test maneuvers involve forceful breathing, which may generate infectious particles. Normal speaking has also been reported to generate small-droplet aerosols, increasing the potential exposure risk in close contact with infected individuals (5–7). Currently, there are no studies evaluating particle generation during pulmonary function tests (PFTs). To better understand the risk associated with PFTs, we sought to quantify and characterize the amount of detectable aerosol and droplet generation during routine pulmonary function studies at prespecified distances. Methods This was a single-center prospective study conducted at the Mayo Clinic in Florida. Five adult volunteer subjects without pulmonary disease consented to perform tidal breathing (Vt), normal speaking, forced vital capacity (FVC), and maximum voluntary ventilation (MVV) maneuvers. The Mayo Clinic Institutional Review Board approved this study (20–005544). Particle measurement A light-scattering particle counter (FLUKE 983) was used to simultaneously measure six channels of particle size distribution (0.3, 0.5, 1, 2, 5, and 10 μm), temperature, and humidity while each maneuver was being performed at each measured location. A minimum sampling volume of 1 L was obtained per measurement. Test conditions To control for humidity, temperature, and other indeterminate ambient variables, room occupancy was limited to three persons (the subject and two investigators) during the study. The pulmonary function test room is 936 cubic feet and has 12.8 room air exchanges per hour. The ambient baseline measurement was obtained at the center of the room before and after each maneuver, with the subject and two investigators in the room and the door closed. The testing room was used only for this study on the day of the testing. The particle counter was positioned as follows: For the speaking portion, the particle counter was positioned 12 inches directly in front of the speaker. For the pulmonary function test portion, the air was sampled at the following three locations: 1) at a 90° angle to and touching the exhalation port (0 ft) on the pulmonary function equipment (Masterscreen PFT; Vyaire Medical), 2) at 1.5 ft from the exhalation port, halfway between where the patient and respiratory therapist sat, and 3) at 3 ft from the exhalation port. The number of particles counted and the volume of air sampled were recorded. The particle counter was zeroed before each test condition. Spirometry was performed outside the plethysmography box. A Microgard II filter (Vyaire Medical) was interposed between the mouthpiece and intake port for all maneuvers, as per manufacturer. For the speaking portion, each subject read “The Rainbow Passage” by Grant Fairbanks for 30 seconds. The subject then performed Vt, FVC, and MVV through the pulmonary function test machine. If the subject coughed during the maneuver, the measurement was discarded, and the study was repeated once ambient particle count had returned to baseline. Each subject performed the test conditions twice. Analysis A nonparametric distribution of measurements was assumed. The Mann-Whitney test for two group comparisons and Wilcoxon/Kruskal-Wallis test for multiple group comparisons was used. If the Kruskal-Wallis test was significant, a Steel test would be performed to compare each group to a single control. This was designated to be the ambient measurement obtained before the respective test. A P value of less than 0.05 was considered significant. Results The five subjects included four men (80%). The mean age was 37.6 ± 8.4 years, the mean height was 177.6 ± 3.9 cm, and the mean weight was 80.4 ± 4.5 kg. Volunteers were white (60%), East Asian Indian (20%), and Asian (20%). Ambient room humidity was measured at 48%, and temperature was 23°C. Table 1 shows the increase in particle counts under different test conditions, with small respirable particles in the 0.3-μm range being generated the most. The FVC and MVV maneuvers generated increased small particles when compared with the ambient measurements with Vt, FVC, and MVV. Table 1. Quantity of particles per liter of sampled air separated by particle size for each maneuver and ambient room measurement Maneuver Particle Quantity Per Liter of Sampled Air by Size 0.3 μm [Mean (95% CI)] 0.5 μm [Mean (95% CI)] 1 μm [Mean (95% CI)] 2 μm [Mean (95% CI)] 5 μm [Mean (95% CI)] 10 μm [Mean (95% CI)] Speaking              Ambient 1,269.6 (1,230.6–1,308.6) 79.5 (51.2–107.8) 21.3 (10.3–32.3) 37.6 (17.3–57.9) 4.9 (3.2–6.6) 3.3 (1.4–5.2)  Speaking for 30 s, 1 ft 1,293.4 (1,242.9–1,343.9) 131.7 (47.3–216.1) 30.2 (23.6–36.8) 47.6 (38.8–56.4) 7.7 (4.0–11.4) 2.7 (0.0–5.4) Vt              Ambient 1,299.2 (1,222.7–1,375.7) 95.2 (80.1–110.3) 33.3 (15.8–50.8) 59.6 (44.0–75.1) 8.9 (5.3–12.5) 4.2 (1.8–6.6)  0 ft 6,466.6(5,991.3–6,941.9) 770.3 (730.5–810.1) 24.7 (16.7–32.7) 21.7 (13.8–29.6) 3.0 (1.4–4.6) 1.4 (0.1–2.7)  1.5 ft 1,250.8 (1,201.8–1,299.8) 88.8 (72.9–104.7) 28.6 (22.5–34.7) 41.6 (36.1–47.1) 5.6 (3.2–8.0) 2.5 (1.0–4.0)  3 ft 1,250.3 (1,190.0–1,310.6) 77.2 (67.3–87.1) 24.2 (17.6–30.8) 32.8 (24.8–40.8) 5.0 (3.4–6.6) 2.2 (1.1–3.3) FVC              Ambient 1,334.8 (1,195.9–1,473.7) 101.8 (71.7–131.9) 31.6 (19.8–43.4) 38.8 (25.4–52.2) 4.7 (1.9–7.5) 2.8 (2.2–3.4)  0 ft 10,739.3 (8,634.7–12,843.9) 1,408.4 (1,076.4–1,740.4) 26.0 (16.1–35.9) 21.1 (12.6–29.6) 2.1 (1.8–2.4) 0.8 (0.2–1.4)  1.5 ft 1,258.2 (1,218.2–1,298.2) 79.0 (73.0–85.0) 23.4 (18.4–28.4) 29.8 (25.0–34.6) 5.0 (2.7–7.3) 1.8 (0.7–2.9)  3 ft 1,266.9 (1,208.3–1,325.5) 73.6 (61.9–85.3) 19.6 (14.2–25.0) 27.0 (22.0–32.0) 2.4 (1.3–3.5) 1.8 (0.4–3.2) MVV              Ambient 1,323.7 (1,226.1–1,421.3) 88.6 (83.3–93.9) 30.8 (14.3–47.3) 40.8 (34.8–46.8) 4.6 (3.9–5.3) 2.4 (1.7–3.1)  0 ft 8,845.2 (6,439.1–11,251.3) 1,251 (444.1–2,057.9) 33.2 (17.5–48.9) 20.8 (14.2–27.4) 3.4 (2.0–4.8) 0.6 (0.0–1.3)  1.5 ft 1,278.1 (1,214.3–1,341.9) 81.0 (58.5–103.5) 29.2 (26.2–32.2) 38.0 (35.5–40.5) 2.8 (1.0–4.6) 2.8 (1.4–4.2)  3 ft 1,283.6 (1,231.2–1,336.0) 82.6 (77.4–87.8) 19.4 (16.2–22.6) 34.6 (29.6–39.6) 4.2 (1.8–6.6) 1.4 (0.0–2.8) Definition of abbreviations: CI = confidence interval; FVC = forced vital capacity; MVV = maximum voluntary ventilation; Vt = tidal volume. Proximity to the source was associated with significant increases in particle generation. With normal speaking, there was not an increase of generated 0.3-μm particles at 12 inches. When sampled close to the exhalation port position, all the maneuvers generated an increase in respirable 0.3-μm particles when compared with baseline (Figure 1). Other aerosol-sized particles measured at 0.5 μm were also increased at this close position. At 1.5 ft and 3 ft from the exhalation port, there was no increase in particle generation for any of the trialed maneuvers. Figure 1. Graph of the concentration of 0.3-μm particles per liter of sampled air generated for each maneuver. P values shown for significant results. FVC = forced vital capacity; MVV = maximum voluntary ventilation. Conclusions This report documents and characterizes aerosol particle generation from routine PFTs and supports precautions to mitigate the transmission of SARS-CoV-2. These precautions include the patient wearing a mask during the instructional phase, maintaining a distance of at least 1.5 ft between the subject and respiratory therapist, using disposable plastic covers over the PFT equipment, and using N95 and personal protective equipment for the respiratory therapist. This study provides valuable information guiding infectious control measures to ensure personnel and patient safety during pulmonary function testing. A limitation of this study was the small sample size of healthy volunteers, which increases the risk of multiple comparisons and decreases the overall stability of the results, but the results were consistent between volunteers, providing reassurance that larger sample sizes may provide similar results. In addition, the high room exchange and humidity may have contributed to particle dispersion and evaporation. Because the volunteers were healthy, the amount and size of particles generated could be considered the minimum, as a patient with nasal secretions or a productive cough may theoretically generate more particles. We did not assess the composition or infectivity of the measured particles, so the possibility of transmission is unknown but possible given the increase in particle generation. In conclusion, PFTs and normal breathing all generate aerosols rather than just droplets. The room air exchange, room turnaround time between testing, and distance between the patient and technician in the testing room are important. Particle generation close to the exhalation port warrants using a single-use plastic covering over the device, with the mouthpiece port and the exhalation port exposed, to avoid equipment contamination. The generation of aerosol-sized particles warrants the use of N95 masks and personal protective equipment during routine PFTs in patients who potentially have an airborne transmissible infectious disease. Supplementary Material Supplements Author disclosures

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          Virological assessment of hospitalized patients with COVID-2019

          Coronavirus disease 2019 (COVID-19) is an acute infection of the respiratory tract that emerged in late 20191,2. Initial outbreaks in China involved 13.8% of cases with severe courses, and 6.1% of cases with critical courses3. This severe presentation may result from the virus using a virus receptor that is expressed predominantly in the lung2,4; the same receptor tropism is thought to have determined the pathogenicity-but also aided in the control-of severe acute respiratory syndrome (SARS) in 20035. However, there are reports of cases of COVID-19 in which the patient shows mild upper respiratory tract symptoms, which suggests the potential for pre- or oligosymptomatic transmission6-8. There is an urgent need for information on virus replication, immunity and infectivity in specific sites of the body. Here we report a detailed virological analysis of nine cases of COVID-19 that provides proof of active virus replication in tissues of the upper respiratory tract. Pharyngeal virus shedding was very high during the first week of symptoms, with a peak at 7.11 × 108 RNA copies per throat swab on day 4. Infectious virus was readily isolated from samples derived from the throat or lung, but not from stool samples-in spite of high concentrations of virus RNA. Blood and urine samples never yielded virus. Active replication in the throat was confirmed by the presence of viral replicative RNA intermediates in the throat samples. We consistently detected sequence-distinct virus populations in throat and lung samples from one patient, proving independent replication. The shedding of viral RNA from sputum outlasted the end of symptoms. Seroconversion occurred after 7 days in 50% of patients (and by day 14 in all patients), but was not followed by a rapid decline in viral load. COVID-19 can present as a mild illness of the upper respiratory tract. The confirmation of active virus replication in the upper respiratory tract has implications for the containment of COVID-19.
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            SARS-CoV-2 Viral Load in Upper Respiratory Specimens of Infected Patients

            To the Editor: The 2019 novel coronavirus (SARS-CoV-2) epidemic, which was first reported in December 2019 in Wuhan, China, and has been declared a public health emergency of international concern by the World Health Organization, may progress to a pandemic associated with substantial morbidity and mortality. SARS-CoV-2 is genetically related to SARS-CoV, which caused a global epidemic with 8096 confirmed cases in more than 25 countries in 2002–2003. 1 The epidemic of SARS-CoV was successfully contained through public health interventions, including case detection and isolation. Transmission of SARS-CoV occurred mainly after days of illness 2 and was associated with modest viral loads in the respiratory tract early in the illness, with viral loads peaking approximately 10 days after symptom onset. 3 We monitored SARS-CoV-2 viral loads in upper respiratory specimens obtained from 18 patients (9 men and 9 women; median age, 59 years; range, 26 to 76) in Zhuhai, Guangdong, China, including 4 patients with secondary infections (1 of whom never had symptoms) within two family clusters (Table S1 in the Supplementary Appendix, available with the full text of this letter at NEJM.org). The patient who never had symptoms was a close contact of a patient with a known case and was therefore monitored. A total of 72 nasal swabs (sampled from the mid-turbinate and nasopharynx) (Figure 1A) and 72 throat swabs (Figure 1B) were analyzed, with 1 to 9 sequential samples obtained from each patient. Polyester flock swabs were used for all the patients. From January 7 through January 26, 2020, a total of 14 patients who had recently returned from Wuhan and had fever (≥37.3°C) received a diagnosis of Covid-19 (the illness caused by SARS-CoV-2) by means of reverse-transcriptase–polymerase-chain-reaction assay with primers and probes targeting the N and Orf1b genes of SARS-CoV-2; the assay was developed by the Chinese Center for Disease Control and Prevention. Samples were tested at the Guangdong Provincial Center for Disease Control and Prevention. Thirteen of 14 patients with imported cases had evidence of pneumonia on computed tomography (CT). None of them had visited the Huanan Seafood Wholesale Market in Wuhan within 14 days before symptom onset. Patients E, I, and P required admission to intensive care units, whereas the others had mild-to-moderate illness. Secondary infections were detected in close contacts of Patients E, I, and P. Patient E worked in Wuhan and visited his wife (Patient L), mother (Patient D), and a friend (Patient Z) in Zhuhai on January 17. Symptoms developed in Patients L and D on January 20 and January 22, respectively, with viral RNA detected in their nasal and throat swabs soon after symptom onset. Patient Z reported no clinical symptoms, but his nasal swabs (cycle threshold [Ct] values, 22 to 28) and throat swabs (Ct values, 30 to 32) tested positive on days 7, 10, and 11 after contact. A CT scan of Patient Z that was obtained on February 6 was unremarkable. Patients I and P lived in Wuhan and visited their daughter (Patient H) in Zhuhai on January 11 when their symptoms first developed. Fever developed in Patient H on January 17, with viral RNA detected in nasal and throat swabs on day 1 after symptom onset. We analyzed the viral load in nasal and throat swabs obtained from the 17 symptomatic patients in relation to day of onset of any symptoms (Figure 1C). Higher viral loads (inversely related to Ct value) were detected soon after symptom onset, with higher viral loads detected in the nose than in the throat. Our analysis suggests that the viral nucleic acid shedding pattern of patients infected with SARS-CoV-2 resembles that of patients with influenza 4 and appears different from that seen in patients infected with SARS-CoV. 3 The viral load that was detected in the asymptomatic patient was similar to that in the symptomatic patients, which suggests the transmission potential of asymptomatic or minimally symptomatic patients. These findings are in concordance with reports that transmission may occur early in the course of infection 5 and suggest that case detection and isolation may require strategies different from those required for the control of SARS-CoV. How SARS-CoV-2 viral load correlates with culturable virus needs to be determined. Identification of patients with few or no symptoms and with modest levels of detectable viral RNA in the oropharynx for at least 5 days suggests that we need better data to determine transmission dynamics and inform our screening practices.
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              SARS-CoV-2 entry factors are highly expressed in nasal epithelial cells together with innate immune genes

              We investigated SARS-CoV-2 potential tropism by surveying expression of viral entry-associated genes in single-cell RNA-sequencing data from multiple tissues from healthy human donors. We co-detected these transcripts in specific respiratory, corneal and intestinal epithelial cells, potentially explaining the high efficiency of SARS-CoV-2 transmission. These genes are co-expressed in nasal epithelial cells with genes involved in innate immunity, highlighting the cells' potential role in initial viral infection, spread and clearance. The study offers a useful resource for further lines of inquiry with valuable clinical samples from COVID-19 patients and we provide our data in a comprehensive, open and user-friendly fashion at www.covid19cellatlas.org.
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                Author and article information

                Journal
                Ann Am Thorac Soc
                Ann Am Thorac Soc
                AnnalsATS
                Annals of the American Thoracic Society
                American Thoracic Society
                2329-6933
                2325-6621
                December 2020
                December 2020
                December 2020
                : 17
                : 12
                : 1637-1639
                Affiliations
                [ 1 ]Mayo Clinic

                Jacksonville, Florida

                and
                [ 2 ]Mayo Clinic

                Rochester, Minnesota
                Author notes
                [* ]Corresponding author (e-mail: helgeson.scott@ 123456mayo.edu ).
                Author information
                http://orcid.org/0000-0001-7590-2293
                http://orcid.org/0000-0003-0891-9495
                Article
                202005-569RL
                10.1513/AnnalsATS.202005-569RL
                7706605
                32870021
                22cd9b38-22cd-4c01-8015-3c4103b7b4b7
                Copyright © 2020 by the American Thoracic Society

                This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 ( http://creativecommons.org/licenses/by-nc-nd/4.0/). For commercial usage and reprints please contact Diane Gern ( dgern@ 123456thoracic.org ).

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