Maosheng Yao

Rapid inactivation of biological species in the air using atmospheric pressure nonthermal plasma.

Here, nonthermal plasma generated by a dielectric barrier discharge (DBD) system was applied to inactivating aerosolized Bacillus subtilis cells and Pseudomonas fluorescens as well as indoor and outdoor bioaerosols. The culturability, viability, and diversity losses of the microorganisms in air samples treated by the plasma for 0.06-0.12 s were studied using culturing, DNA stain as well as polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) methods. In addition, the viable fraction of bacterial aerosols with and without the plasma treatment was also quantified using qPCR coupled with ethidium monoazide (EMA). It was shown that less than 2% of B. subtilis aerosols survived the plasma treatment of 0.12 s, while none of the P. fluorescens aerosols survived. Viability tests, EMA-qPCR results, and Scanning Electron Microscopy (SEM) images demonstrated that both bacterial species suffered significant viability loss, membrane, and DNA damages. Exposure of environmental bacterial and fungal aerosols to the plasma for 0.06 s also resulted in their significant inactivations, more than 95% for bacteria and 85-98% for fungal species. PCR-DGGE analysis showed that plasma exposure of 0.06 s resulted in culturable bacterial aerosol diversity loss for both environments, especially pronounced for indoor environment. The results here demonstrate that nonthermal plasma exposure could offer a highly efficient air decontamination technology.

Characterization of Biological Aerosol Exposure Risks from Automobile Air Conditioning System

Although use of automobile air conditioning (AC) was shown to reduce in-vehicle particle levels, the characterization of its microbial aerosol exposure risks is lacking. Here, both AC and engine filter dust samples were collected from 30 automobiles in four different geographical locations in China. Biological contents (bacteria, fungi, and endotoxin) were studied using culturing, high-throughput gene sequence, and Limulus amebocyte lysate (LAL) methods. In-vehicle viable bioaerosol concentrations were directly monitored using an ultraviolet aerodynamic particle sizer (UVAPS) before and after use of AC for 5, 10, and 15 min. Regardless of locations, the vehicle AC filter dusts were found to be laden with high levels of bacteria (up to 26,150 CFU/mg), fungi (up to 1287 CFU/mg), and endotoxin (up to 5527 EU/mg). More than 400 unique bacterial species, including human opportunistic pathogens, were detected in the filter dusts. In addition, allergenic fungal species were also found abundant. Surprisingly, unexpected fluorescent peaks around 2.5 μm were observed during the first 5 min use of AC, which was attributed to the reaerosolization of those filter-borne microbial agents. The information obtained here can assist in minimizing or preventing the respiratory allergy or infection risk from the use of automobile AC system.

Integrating silicon nanowire field effect transistor, microfluidics and air sampling techniques for real-time monitoring biological aerosols.

Numerous threats from biological aerosol exposures, such as those from H1N1 influenza, SARS, bird flu, and bioterrorism activities necessitate the development of a real-time bioaerosol sensing system, which however is a long-standing challenge in the field. Here, we developed a real-time monitoring system for airborne influenza H3N2 viruses by integrating electronically addressable silicon nanowire (SiNW) sensor devices, microfluidics and bioaerosol-to-hydrosol air sampling techniques. When airborne influenza H3N2 virus samples were collected and delivered to antibody-modified SiNW devices, discrete nanowire conductance changes were observed within seconds. In contrast, the conductance levels remained relatively unchanged when indoor air or clean air samples were delivered. A 10-fold increase in virus concentration was found to give rise to about 20-30% increase in the sensor response. The selectivity of the sensing device was successfully demonstrated using H1N1 viruses and house dust allergens. From the simulated aerosol release to the detection, we observed a time scale of 1-2 min. Quantitative polymerase chain reaction (qPCR) tests revealed that higher virus concentrations in the air samples generally corresponded to higher conductance levels in the SiNW devices. In addition, the display of detection data on remote platforms such as cell phone and computer was also successfully demonstrated with a wireless module. The work here is expected to lead to innovative methods for biological aerosol monitoring, and further improvements in each of the integrated elements could extend the system to real world applications.

MS2 Virus Inactivation by Atmospheric-Pressure Cold Plasma Using Different Gas Carriers and Power Levels

ABSTRACT In this study, airborne MS2 bacteriophages were exposed for subsecond time intervals to atmospheric-pressure cold plasma (APCP) produced using different power levels (20, 24, and 28 W) and gas carriers (ambient air, Ar-O2 [2%, vol/vol], and He-O2 [2%, vol/vol]). In addition, waterborne MS2 viruses were directly subjected to the APCP treatment for up to 3 min. MS2 viruses with and without the APCP exposure were examined by scanning electron microscopy (SEM), reverse transcription-PCR (RT-PCR), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Viral inactivation was shown to exhibit linear relationships with the APCP generation power and exposure time (R 2 > 0.95 for all energy levels tested) up to 95% inactivation (1.3-log reduction) after a subsecond airborne exposure at 28 W; about the same inactivation level was achieved for waterborne viruses with an exposure time of less than 1 min. A larger amount of reactive oxygen species (ROS), such as atomic oxygen, in APCP was detected for a higher generation power with Ar-O2 and He-O2 gas carriers. SEM images, SDS-PAGE, and agarose gel analysis of exposed waterborne viruses showed various levels of damage to both surface proteins and their related RNA genes after the APCP exposure, thus leading to the loss of their viability and infectivity.

Ambient bioaerosol particle dynamics observed during haze and sunny days in Beijing.

The chemical characteristics of airborne particulate matter (PM) have been extensively studied; however, little information exists for its biological components (bioaerosol) especially during a haze event in mega cities. Herein, we studied the bioaerosol (fluorescent particle) dynamics on both haze and sunny days in Beijing from Dec. 2013 to March 2014 by employing a widely used real-time bioaerosol sensor-ultraviolet aerodynamic particle spectrometer (UV-APS). Firstly, we studied the fluorescent particle (BioPM) concentration and size distributions during three independent haze and three independent sunny days. Secondly, we investigated BioPM dynamics over a two-week long monitoring period which included consecutive haze days and alternated sunny days. In addition, we analyzed bacterial community structures and endotoxin levels in the air samples using pyrosequencing and Limulus amebocyte lysate (LAL) method, respectively. More than 6-fold higher fluorescent particle concentrations up to 5×10(5)/m(3) with peaks at night or early dawn were detected at the time of haze occurrences than those observed on sunny days. When the haze episode progressed for 3-5days, the BioPM concentrations were observed to decrease to the levels that were typically observed on sunny days. In general, ozone levels were found to be elevated at noon, while BioPM, NOx and relative humidity were reduced. Gene sequence analysis revealed no significant difference in abundances and community structures for top 13 bacterial genera between haze and sunny days, yet about twice higher endotoxin levels (12.4EU/m(3)) were detected on haze days than on sunny days. The results here facilitate a better understanding of atmospheric fluorescent particle dynamics including those under haze events.

The allergenicity of Aspergillus fumigatus conidia is influenced by growth temperature.

Common indoor and outdoor environmental fungi such as Aspergillus fumigatus produce asexual spores containing a collection of proteins that can bind IgE antibodies and trigger allergic reactions. We characterized the impact of sporulation temperature on the IgE-binding capacity (allergenicity) of A. fumigatus and explored the links between variable allergenicityxa0and temperature-dependant expression of genes encoding these allergenic proteins. A 12-fold increase in A. fumigatus allergenicity per spore was observed when sporulation temperatures were decreased from 32°C to 17°C. Per spore protein mass and Asp f 1 allergen mass also followed this trend. Functional gene expression analysis of A. fumigatus sporulating cultures by real-time reverse-transcription PCR and gene expression microarrays revealed that a greater number of genes encoding known, major allergens are more highly expressed at lower sporulation temperatures. The results of this study indicate that environmental conditions at growth significantly influence the allergenicity of this common mould through the differential production of allergenic proteins, and highlight the importance of in vivo or in vitro allergenicity measurements for understanding environmental exposure to airborne allergenic fungi.

Inactivation and Magnetic Separation of Bacteria from Liquid Suspensions Using Electrosprayed and Nonelectrosprayed nZVI Particles: Observations and Mechanisms

Here, nonelectrosprayed nanoscale zerovalent iron (NE-nZVI), electrosprayed nZVI (E-nZVI) and preoxidized nZVI (O-nZVI) particles were applied to inactivating Bacillus subtilis, Escherichia coli as well as bacteria in various wastewater samples. In addition, magnetic separation was applied to the mixture of 0.2 mL bacterial sample and 1.8 mL E-nZVI or NE-nZVI suspensions. Bacterial concentrations and optical density of the supernatants were analyzed using culturing, optical adsorption and qPCR tests. In general, for wastewater samples the inactivations were shown to range from 1-log to 3-log. PCR-DGGE analysis indicated that no gene mutation occurred when bacteria were treated with nZVI. Using magnetic separation, significant physical removals, revealed as a function of nZVI type (NE-,E- and O-nZVI) and bacterial concentration, up to 6-log were obtained. E-nZVI and NE-nZVI were shown to react differently with B. subtilis and E. coli, although exhibiting similar inactivation rates. qPCR tests detected higher amount of DNA in the supernatants from mixing E. coli with NE-nZVI, but less for E-nZVI. However, the opposite was observed with B. subtilis. Our data together with optical adsorption analysis suggested that the inactivation and magnetic separation mainly depend on Fe(0)/Fe(3)O(4) shell compositions, the type of bacteria (aerobic and anaerobic) and their concentrations.

Rapid allergen inactivation using atmospheric pressure cold plasma.

Summary form only given. Exposure to environmental allergens from various sources can cause a wide range of adverse health effects, including asthma, eczema, allergic rhinitis, inflammable and toxic reactions. Among them, asthma is one of the most common and major human diseases that are attributable to allergen exposure. And such a problem continues to rise e.g., the asthma prevalence among children in the United States alone was indicated to have nearly doubled over the last two decades. Allergy has become a global problem, and effective control is greatly needed. Here, the inactivation effects of the atmospheric pressure cold plasma (APCP) on aerosolized allergens including dog allergen Can f 1, house dust mite allergens (Der p 1 and Der f 1), fungal allergens (Asp f 1 and Alt a 1) as well as those from indoor and outdoor environments were investigated. The effectiveness of the APCP treatment was further studied using blood sera from the allergen sensitized humans. In addition, the allergen samples were also analyzed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results revealed that the APCP was highly effective in reducing the allergenicity of both lab-prepared and environmental allergen aerosols. The airborne reductions were shown to range from 30% for Der p 1 to 80% for Can f 1 allergen for 0.12 s exposure. Allergnicity tests showed that the APCP treated Asp f 1 allergens caused 50% less binding with IgEs in the blood sera compared to the control. The observed allergenicity loss was due to hydroxyl radicals produced by the plasma device. The results from SDS-PAGE showed that the plasma treatment resulted in decreased size of the Asp f 1 allergen. The developed technology holds great promise in combating the allergic diseases.

Rapid allergen inactivation using atmospheric pressure cold plasma

Exposure to environmental allergens from various sources can cause a wide range of adverse health effects, including asthma, eczema, allergic rhinitis, inflammable and toxic reactions. Among them, asthma is one of the most common and major human diseases that are attributable to allergen exposure. And such a problem continues to rise e.g., the asthma prevalence among children in the United States alone was indicated to have nearly doubled over the last two decades. Allergy has become a global problem, and effective control is greatly needed.

In situ airborne virus inactivation by microwave irradiation

Infectious diseases cause tremendous costs of both human and economy annually. Previously, we have studied the bacterial, fungal, and allergen aerosol inactivation by direct microwave irradiation. Here, we further investigated its effects on airborne viruses. MS2 coliphage used as a human model virus was aerosolized and exposed to the direct microwave irradiation for ~2xa0min at three different power levels (700, 385, and 119xa0W). In addition to the survival rate, the viral genes before and after the microwave treatments were also examined using PCR and gel electrophoresis. Direct exposure of airborne MS2 viruses to the microwave irradiation at 700xa0W for less than 2xa0min was shown to result in more than 90xa0% inactivation efficiency, about 65xa0% at medium power level (385xa0W), and 50xa0% at the lowest level (119xa0W). The aerosol inactivation rate followed a linear relationship with the microwave exposure time (R2xa0=xa00.9889). Scanning electron images revealed visible damages to the viral surface after the exposure. Damages were also observed to the viral RNA genes coding for coat proteins, among which the A protein gene was completely destroyed. This study demonstrated that even without the filtration the direct microwave irradiation could also achieve rapid inactivation of viral aerosols. The information obtained can provide useful guidance on the development of microwave-based viral threat mitigation solutions in a closed or semi-closed space.