A quanta-independent approach for the assessment of strategies to reduce the risk of airborne infection.

The Wells-Riley model is extensively used for retrospective and prospective modelling of the risk of airborne transmission of infection in indoor spaces. It is also used when examining the efficacy of various removal and deactivation methods for airborne infectious aerosols in the indoor environment, which is crucial when selecting the most effective infection control technologies. The problem is that the large variation in viral load between individuals makes the Wells-Riley model output very sensitive to the input parameters and may yield a flawed prediction of risk. The absolute infection risk estimated with this model can range from nearly 0 % to 100 % depending on the viral load, even when all other factors, such as removal mechanisms and room geometry, remain unchanged. We therefore propose a novel method that removes this sensitivity to viral load. We define a quanta-independent maximum absolute before-after difference in infection risk that is independent of quanta factors like viral load, physical activity, or the dose-response relationships. The input data needed for a non-steady-state calculation are just the removal rates, room volume, and occupancy duration. Under steady-state conditions the approach provides an elegant solution that is only dependent on removal mechanisms before and after applying infection control measures. We applied this method to compare the impact of relative humidity, ventilation rate and its effectiveness, filtering efficiency, and the use of ultraviolet germicidal irradiation on the infection risk. The results demonstrate that the method provides a comprehensive understanding of the impact of infection control strategies on the risk of airborne infection, enabling rational decisions to be made regarding the most effective strategies in a specific context. The proposed method thus provides a practical tool for mitigation of airborne infection risk.

Mandating indoor air quality for public buildings.

If some countries lead by example, standards may increasingly become normalized.

Toxicity Screening of Fungal Extracts and Metabolites, Xenobiotic Chemicals, and Indoor Dusts with In Vitro and Ex Vivo Bioassay Methods.

It is controversial how useful bioassays are for identifying the in vivo toxicity of hazardous environmental exposures. In this study, fruiting bodies of forest mushrooms (n = 46), indoor mold colonies (n = 412), fungal secondary metabolites (n = 18), xenobiotic chemicals such as biocides and detergents (n = 6), and methanol extracts of indoor dusts from urban buildings (n = 26) were screened with two different bioactivity assays: boar sperm motility inhibition (BSMI) and inhibition of cell proliferation (ICP) tests. For the forest mushrooms, the toxicity testing result was positive for 100% of poisonous-classified species, 69% of non-edible-classified species, and 18% of edible-classified species. Colonies of 21 isolates of Ascomycota mold fungal species previously isolated from water-damaged buildings proved to be toxic in the tests. Out of the fungal metabolites and xenobiotic chemicals, 94% and 100% were toxic, respectively. Out of the indoor dusts from moldy-classified houses (n = 12) and from dry, mold-free houses (n = 14), 50% and 57% were toxic, respectively. The bioassay tests, however, could not differentiate the samples from indoor dusts of moldy-classified buildings from those from the mold-free buildings. Xenobiotic chemicals and indoor dusts were more toxic in the BSMI assay than in the ICP assay, whereas the opposite results were obtained with the Ascomycota mold colonies and fungal secondary metabolites. The tests recognized unknown methanol-soluble thermoresistant substances in indoor settled dusts. Toxic indoor dusts may indicate a harmful exposure, regardless of whether the toxicity is due to xenobiotic chemicals or microbial metabolites.

Coronavirus Disease 2019 and Airborne Transmission: Science Rejected, Lives Lost. Can Society Do Better?

This is an account that should be heard of an important struggle: the struggle of a large group of experts who came together at the beginning of the COVID-19 pandemic to warn the world about the risk of airborne transmission and the consequences of ignoring it. We alerted the World Health Organization about the potential significance of the airborne transmission of SARS-CoV-2 and the urgent need to control it, but our concerns were dismissed. Here we describe how this happened and the consequences. We hope that by reporting this story we can raise awareness of the importance of interdisciplinary collaboration and the need to be open to new evidence, and to prevent it from happening again. Acknowledgement of an issue, and the emergence of new evidence related to it, is the first necessary step towards finding effective mitigation solutions.

New dose-response model and SARS-CoV-2 quanta emission rates for calculating the long-range airborne infection risk.

Predictive models for airborne infection risk have been extensively used during the pandemic, but there is yet still no consensus on a common approach, which may create misinterpretation of results among public health experts and engineers designing building ventilation. In this study we applied the latest data on viral load, aerosol droplet sizes and removal mechanisms to improve the Wells Riley model by introducing the following novelties i) a new model to calculate the total volume of respiratory fluid exhaled per unit time ii) developing a novel viral dose-based generation rate model for dehydrated droplets after expiration iii) deriving a novel quanta-RNA relationship for various strains of SARS-CoV-2 iv) proposing a method to account for the incomplete mixing conditions. These new approaches considerably changed previous estimates and allowed to determine more accurate average quanta emission rates including omicron variant. These quanta values for the original strain of 0.13 and 3.8 quanta/h for breathing and speaking and the virus variant multipliers may be used for simple hand calculations of probability of infection or with developed model operating with six size ranges of aerosol droplets to calculate the effect of ventilation and other removal mechanisms. The model developed is made available as an open-source tool.

Zonal modeling of air distribution impact on the long-range airborne transmission risk of SARS-CoV-2.

A widely used analytical model to quantitatively assess airborne infection risk is the Wells-Riley model which is limited to complete air mixing in a single zone. However, this assumption tends not to be feasible (or reality) for many situations. This study aimed to extend the Wells-Riley model so that the infection risk can be calculated in spaces where complete mixing is not present. Some more advanced ventilation concepts create either two horizontally divided air zones in spaces as displacement ventilation or the space may be divided into two vertical zones by downward plane jet as in protective-zone ventilation systems. This is done by evaluating the time-dependent distribution of infectious quanta in each zone and by solving the coupled system of differential equations based on the zonal quanta concentrations. This model introduces a novel approach by estimating the interzonal mixing factor based on previous experimental data for three types of ventilation systems: incomplete mixing ventilation, displacement ventilation, and protective zone ventilation. The modeling approach is applied to a room with one infected and one susceptible person present. The results show that using the Wells-Riley model based on the assumption of completely air mixing may considerably overestimate or underestimate the long-range airborne infection risk in rooms where air distribution is different than complete mixing, such as displacement ventilation, protected zone ventilation, warm air supplied from the ceiling, etc. Therefore, in spaces with non-uniform air distribution, a zonal modeling approach should be preferred in analytical models compared to the conventional single-zone Wells-Riley models when assessing long-range airborne transmission risk of infectious respiratory diseases.

Modeling the impact of indoor relative humidity on the infection risk of five respiratory airborne viruses.

With a modified version of the Wells-Riley model, we simulated the size distribution and dynamics of five airborne viruses (measles, influenza, SARS-CoV-2, human rhinovirus, and adenovirus) emitted from a speaking person in a typical residential setting over a relative humidity (RH) range of 20-80% and air temperature of 20-25 °C. Besides the size transformation of virus-containing droplets due to evaporation, respiratory absorption, and then removal by gravitational settling, the modified model also considered the removal mechanism by ventilation. The trend and magnitude of RH impact depended on the respiratory virus. For rhinovirus and adenovirus humidifying the indoor air from 20/30 to 50% will be increasing the relative infection risk, however, this relative infection risk increase will be negligible for rhinovirus and weak for adenovirus. Humidification will have a potential benefit in decreasing the infection risk only for influenza when there is a large infection risk decrease for humidifying from 20 to 50%. Regardless of the dry solution composition, humidification will overall increase the infection risk via long-range airborne transmission of SARS-CoV-2. Compared to humidification at a constant ventilation rate, increasing the ventilation rate to moderate levels 0.5 → 2.0 h-1 will have a more beneficial infection risk decrease for all viruses except for influenza. Increasing the ventilation rate from low values of 0.5 h-1 to higher levels of 6 h-1 will have a dominating effect on reducing the infection risk regardless of virus type.

Respiratory infection risk-based ventilation design method.

A new design method is proposed to calculate outdoor air ventilation rates to control respiratory infection risk in indoor spaces. We propose to use this method in future ventilation standards to complement existing ventilation criteria based on the perceived air quality and pollutant removal. The proposed method makes it possible to calculate the required ventilation rate at a given probability of infection and quanta emission rate. Present work used quanta emission rates for SARS-CoV-2 and consequently the method can be applied for other respiratory viruses with available quanta data. The method was applied to case studies representing typical rooms in public buildings. To reduce the probability of infection, the total airflow rate per infectious person revealed to be the most important parameter to reduce the infection risk. Category I ventilation rate prescribed in the EN 16798-1 standard satisfied many but not all type of spaces examined. The required ventilation rates started from about 80 L/s per room. Large variations between the results for the selected case studies made it impossible to provide a simple rule for estimating the required ventilation rates. Consequently, we conclude that to design rooms with a low infection risk the newly developed ventilation design method must be used.

Estimating the impact of indoor relative humidity on SARS-CoV-2 airborne transmission risk using a new modification of the Wells-Riley model.

A novel modified version of the Wells-Riley model was used to estimate the impact of relative humidity (RH) on the removal of respiratory droplets containing the SARS-CoV-2 virus by deposition through gravitational settling and its inactivation by biological decay; the effect of RH on susceptibility to SARS-CoV-2 was not considered. These effects were compared with the removal achieved by increased ventilation rate with outdoor air. Modeling was performed assuming that the infected person talked continuously for 60 and 120 min. The results of modeling showed that the relative impact of RH on the infection risk depended on the ventilation rate and the size range of virus-laden droplets. A ventilation rate of 0.5 ACH, the change of RH between 20% and 53% was predicted to have a small effect on the infection risk, while at a ventilation rate of 6 ACH this change had nearly no effect. On the contrary, increasing the ventilation rate from 0.5 ACH to 6 ACH was predicted to decrease the infection risk by half which is remarkably larger effect compared with that predicted for RH. It is thus concluded that increasing the ventilation rate is more beneficial for reducing the airborne levels of SARS-CoV-2 than changing indoor RH. PRACTICAL IMPLICATIONS: The present results show that humidification to moderate levels of 40%-60% RH should not be expected to provide a significant reduction in infection risk caused by SARS-CoV-2, hence installing and running humidifiers may not be an efficient solution to reduce the risk of COVID-19 disease in indoor spaces. The results do however confirm that ventilation has a key role in controlling SARS-CoV-2 virus concentration in the air providing considerably higher benefits. The modified model developed in the present work can be used by public health experts, engineers, and epidemiologists when selecting different measures to reduce the infection risk from SARS-CoV-2 indoors allowing informed decisions concerning indoor environmental control.

Melinacidin-Producing Acrostalagmus luteoalbus, a Major Constituent of Mixed Mycobiota Contaminating Insulation Material in an Outdoor Wall.

Occupants may complain about indoor air quality in closed spaces where the officially approved standard methods for indoor air quality risk assessment fail to reveal the cause of the problem. This study describes a rare genus not previously detected in Finnish buildings, Acrostalagmus, and its species A. luteoalbus as the major constituents of the mixed microbiota in the wet cork liner from an outdoor wall. Representatives of the genus were also present in the settled dust in offices where occupants suffered from symptoms related to the indoor air. One strain, POB8, was identified as A. luteoalbus by ITS sequencing. The strain produced the immunosuppressive and cytotoxic melinacidins II, III, and IV, as evidenced by mass spectrometry analysis. In addition, the classical toxigenic species indicating water damage, mycoparasitic Trichoderma, Aspergillus section Versicolores, Aspergillus section Circumdati, Aspergillus section Nigri, and Chaetomium spp., were detected in the wet outdoor wall and settled dust from the problematic rooms. The offices exhibited no visible signs of microbial growth, and the airborne load of microbial conidia was too low to explain the reported symptoms. In conclusion, we suggest the possible migration of microbial bioactive metabolites from the wet outdoor wall into indoor spaces as a plausible explanation for the reported complaints.

Transmission of SARS-CoV-2 by inhalation of respiratory aerosol in the Skagit Valley Chorale superspreading event.

During the 2020 COVID-19 pandemic, an outbreak occurred following attendance of a symptomatic index case at a weekly rehearsal on 10 March of the Skagit Valley Chorale (SVC). After that rehearsal, 53 members of the SVC among 61 in attendance were confirmed or strongly suspected to have contracted COVID-19 and two died. Transmission by the aerosol route is likely; it appears unlikely that either fomite or ballistic droplet transmission could explain a substantial fraction of the cases. It is vital to identify features of cases such as this to better understand the factors that promote superspreading events. Based on a conditional assumption that transmission during this outbreak was dominated by inhalation of respiratory aerosol generated by one index case, we use the available evidence to infer the emission rate of aerosol infectious quanta. We explore how the risk of infection would vary with several influential factors: ventilation rate, duration of event, and deposition onto surfaces. The results indicate a best-estimate emission rate of 970 ± 390 quanta/h. Infection risk would be reduced by a factor of two by increasing the aerosol loss rate to 5 h-1 and shortening the event duration from 2.5 to 1 h.

Bioreactivity, Guttation and Agents Influencing Surface Tension of Water Emitted by Actively Growing Indoor Mould Isolates.

The secretion of metabolites in guttation droplets by indoor moulds is not well documented. This study demonstrates the guttation of metabolites by actively growing common indoor moulds. Old and fresh biomasses of indoor isolates of Aspergillus versicolor, Chaetomium globosum, Penicillium expansum, Trichoderma atroviride, T. trixiae, Rhizopus sp. and Stachybotrys sp. were compared. Metabolic activity indicated by viability staining and guttation of liquid droplets detected in young (<3 weeks old) biomass were absent in old (>6 months old) cultures consisting of dehydrated hyphae and dormant conidia. Fresh (<3 weeks old) biomasses were toxic more than 10 times towards mammalian cell lines (PK-15 and MNA) compared to the old dormant, dry biomasses, when calculated per biomass wet weight and per conidial particle. Surfactant activity was emitted in exudates from fresh biomass of T. atroviride, Rhizopus sp. and Stachybotrys sp. Surfactant activity was also provoked by fresh conidia from T. atroviride and Stachybotrys sp. strains. Water repealing substances were emitted by cultures of P. expansum, T. atroviride and C. globosum strains. The metabolic state of the indoor fungal growth may influence emission of liquid soluble bioreactive metabolites into the indoor air.

Detection of Chaetomium globosum, Ch. cochliodes and Ch. rectangulare during the Diversity Tracking of Mycotoxin-Producing Chaetomium-Like Isolates Obtained in Buildings in Finland.

The diversity of Chaetomium-like isolates in buildings in Finland is poorly documented. This paper describes a set of methods for rapid diversity tracking of 42 indoor Chaetomium-like isolates. These isolates were categorized based on their fluorescence emission, ascomatal hair morphology, responses in three bioassays and resistance/sensitivity to the wetting agent Genapol X-080. Thirty-nine toxigenic isolates were identified [Ch. globosum (n = 35), Ch. cochliodes (n = 2) and Ch. rectangulare (n = 2)]. These isolates were identified down to the species level by tef1α gene sequencing. The major toxic substances in the ethanol extracts of the Ch. globosum and Ch. cochliodes strains were chaetoglobosin, chaetoviridin A and C, chaetomugilin D and chaetomin, identified based on HPLC-UV and mass spectrometry data (MS and MS/MS). Ethanol extracts from pure Ch. globosum cultures exhibited a toxicological profile in the boar sperm motility inhibition assay (BSMI), sperm membrane integrity damage assay (SMID) and inhibition of cell proliferation (ICP) assay, similar to that exhibited by pure chaetoglobosin A. Overall, differences in fluorescence, morphology, toxicity profile, mycotoxin production and sensitivity to chemicals were consistent with those in tef1α sequencing results for species identification. The results indicate the presence of Ch. cochliodes and Ch. rectangulare in Finnish buildings, representing a new finding.

How can airborne transmission of COVID-19 indoors be minimised?

During the rapid rise in COVID-19 illnesses and deaths globally, and notwithstanding recommended precautions, questions are voiced about routes of transmission for this pandemic disease. Inhaling small airborne droplets is probable as a third route of infection, in addition to more widely recognized transmission via larger respiratory droplets and direct contact with infected people or contaminated surfaces. While uncertainties remain regarding the relative contributions of the different transmission pathways, we argue that existing evidence is sufficiently strong to warrant engineering controls targeting airborne transmission as part of an overall strategy to limit infection risk indoors. Appropriate building engineering controls include sufficient and effective ventilation, possibly enhanced by particle filtration and air disinfection, avoiding air recirculation and avoiding overcrowding. Often, such measures can be easily implemented and without much cost, but if only they are recognised as significant in contributing to infection control goals. We believe that the use of engineering controls in public buildings, including hospitals, shops, offices, schools, kindergartens, libraries, restaurants, cruise ships, elevators, conference rooms or public transport, in parallel with effective application of other controls (including isolation and quarantine, social distancing and hand hygiene), would be an additional important measure globally to reduce the likelihood of transmission and thereby protect healthcare workers, patients and the general public.

Exposure to indoor air contaminants in school buildings with and without reported indoor air quality problems.

Reported indoor air quality (IAQ) complaints are common even in relatively new or renovated school buildings in Finland. However, detecting the causes for complaints with commonly used indoor air measurements is difficult. This study presents data on perceived and measured IAQ in six comprehensive school buildings in Finland. The aim of this study was to discover the possible differences of perceived and measured IAQ between schools with reported IAQ complaints and schools without reported IAQ complaints. The initial categorisation of schools with ('problematic schools') and without ('comparison schools') complaints was ensured via a validated indoor climate survey and a recently developed online questionnaire, which were completed by 186 teachers and 1268 students from the six schools. IAQ measurements of physical parameters, gaseous pollutants, particulate matter and bioaerosols were conducted in four problematic school buildings (26 classrooms) and two comparison school buildings (12 classrooms). Using air sampling as well as exhaust air filters and classroom settled dust to detect the presence of elevated concentrations of airborne cultivable microbes and pathogenic, toxigenic and mycoparasitic Trichoderma strains were the most indicative methods in distinguishing problematic schools from comparison schools. Other IAQ-related measurements did not detect clear differences between problematic and comparison schools, as the concentration levels were very low. The results indicate that the complaints reported by occupants could have been related to excess moisture or mould problems that had not been found or repaired. Ventilation pressure condition investigations and simultaneous exhaust and supply air filter dust culture should be addressed precisely in future studies.

Screening Mold Colonies by Using Two Toxicity Assays Revealed Indoor Strains of Aspergillus calidoustus Producing Ophiobolins G and K.

The occurrence and toxin production of the opportunistic pathogen Aspergillus calidoustus in Finnish buildings is not well documented in the literature. We tracked and identified four A. calidoustus colonies cultivated from indoor settled dusts and revealed the biological activities of crude biomass extracts. The toxic substances were identified as 6-epi-ophiobolin K, ophiobolin K, and ophiobolin G by high-performance liquid chromatography-mass spectrometry (HPLC-MS) based on chromatographic and mass spectrometry data (MS and MS/MS) on the crude extract of A. calidoustus strain MH34. A total of 29 fungal colonies collected from settled dust in an office room reported for indoor-air-related illnesses were screened for toxins that inhibited boar sperm motility in the BSMI (boar sperm motility inhibiting) assay and cell proliferation in the ICP (inhibition of cell proliferation) assays with PK-15 cells. Out of the 27 colonies tested as toxic, 12 colonies exhibiting conidiophores representative of the genera Chaetomium, Penicillium, and Paecilomyces were excluded from the study, while 13 colonies exhibited Aspergillus-like conidiophores. Biomass suspensions of these colonies were divided into two categories: Category 1 colonies (n = 4), toxic in the BSMI assay and the ICP assays, emitted blue fluorescence and grew at 37 °C; Category 2 colonies (n = 9), only toxic in the ICP assay, emitted orange fluorescence and exhibited limited or no growth at 37 °C. Colonies in Category 1 were pure-cultured, and the strains were named as MH4, MH21, MH34, MH36. Strain MH34 was identified as A. calidoustus by the internal transcribed spacer (ITS) sequences. Ethanol-soluble dry substances extracted from the biomass of the pure cultures exhibited a toxicological profile in the BSMI assay, SMID (sperm membrane integrity damage) assay, and ICP assay similar to that exhibited by pure ophiobolin A. Overall, the viable conidia of A. calidoustus in indoor settled dusts deserve attention when potentially hazardous mold species are monitored.

Emissions of DEHP-free PVC flooring.

Degrading 2-ethylhexyl-containing PVC floorings (eg DEHP-PVC floorings) and adhesives emit 2-ethylhexanol (2-EH) in the indoor air. The danger of flooring degradation comes from exposing occupants to harmful phthalates plasticisers (eg DEHP), but not from 2-EH as such. Since the EU banned the use of phthalates in sensitive applications, the market is shifting to use DEHP-free and alternative types of plasticisers in PVC products. However, data on emissions from DEHP-free PVC floorings are scarce. This study aimed at assessing the surface and bulk emissions of two DEHP-free PVC floorings over three years. The floorings were glued on the screed layer of concrete casts at 75%, 85%, and 95% RH. The volatile organic compounds (VOCs) were actively sampled using FLEC (surface emissions) and micro-chamber/thermal extractor (µ-CTE, bulk emissions) onto Tenax TA adsorbents and analyzed with TD-GC-MS. 2-EH, C9-alcohols, and total volatile organic compound (TVOC) emissions are reported. Emissions at 75% and 85% RH were similar. As expected, the highest emissions occurred at 95% RH. 2-EH emissions originated from the adhesive. Because the two DEHP-free floorings tested emitted C9-alcohols at all tested RH, it makes the detection of flooring degradation harder, particularly if the adhesive used does not emit 2-EH.

An Evaluation of Boar Spermatozoa as a Biosensor for the Detection of Sublethal and Lethal Toxicity.

A novel, objective, and rapid computed motility inhibition (CMI) assay was developed to identify and assess sublethal injury in toxin-exposed boar spermatozoa and compared with a subjective visual motility inhibition (VMI) assay. The CMI values were calculated from digital micrographic videos using a custom MATLAB® script by contrasting the motility index values of each experiment with those of the background and control experiments. Following a comparison of the CMI and VMI assays results, it was determined that their agreement depended on the shape of the dose-response curve. Toxins that exhibited a steep slope were indicative of good agreement between the assays. Those depicted by a gentle decline in the slope of the dose-response curve, the CMI assay were shown to be two times more sensitive than the VMI assay. The CMI assay was highly sensitive to the inhibition of mitochondrial function and glucose transport activity by sublethal doses of toxins and to disruption of cellular cation homeostasis by carrier ionophoric toxins, when compared to the cytotoxicity and lethal toxicity assays (i.e., that evaluated the inhibition of cell proliferation in somatic cell lines (FL, PK-15, and MNA cells)) and disruption to spermatozoa membrane integrity. The CMI assay recognized subtle sublethal toxicity changes in metabolism, manifested as a decrease in boar spermatozoa motility. Thus, it was feasible to effectively compare the objectively-measured numerical values for motility inhibition using the CMI assay against those reflecting lethal damage in the spermatozoa cells and somatic cell lines using a cytotoxicity assay.