A new personal bioaerosol sampler has recently been developed and evaluated for sampling of viable airborne bacteria and fungi under controlled laboratory conditions and in the field. The operational principle of the device is based on the passage of air through porous medium immersed in liquid. This process leads to the formation of bubbles within the filter as the carrier gas passes through and thus provides effective mechanisms for aerosol removal. As demonstrated in previous studies, the culturability of sampled bacterium and fungi remained high for the entire 8-h sampling period. The present study is the first step of the evaluation of the new sampler for monitoring of viable airborne viruses. It focuses on the investigation of the inactivation rate of viruses in the bubbling process during 4 h of continuous operation. Four microbes were used in this study, influenza, measles, mumps, and vaccinia viruses. It was found that the use of distilled water as the collection fluid was associated with a relatively high decay rate. A significant improvement was achieved by utilizing virus maintenance fluid prepared by using Hank's solution with appropriate additives. The survival rates of the influenza, measles, and mumps viruses were increased by 1.4 log, 0.83 log, and 0.82 log, respectively, after the first hour of operation compared to bubbling through the sterile water. The same trend was observed throughout the entire 4-h experiment. There was no significant difference observed only for the robust vaccinia virus.

Spread of Venezuelan equine encephalitis (VEE) virus and damage of the central nervous system (CNS) in mice infected by respiratory route was studied. Virus concentration in organs and blood, "dose-effect" relationships, and ultrastructural lesions in various tissues were examined in immune and normal mice. We showed, via three independent methods--characteristic curve investigations, tissue virus concentration dynamics, and ultrastructural methods--the spread of VEE virus through the olfactory tract into the brain of immune mice. From these experiments it was concluded that in case of respiratory challenge VEE virus can enter the CNS of normal mice by both vascular and olfactory pathways, while in immune mice the main route is olfactory.

While various sampling methods exist for collecting and enumerating airborne bacteria and fungi, no credible methodology has yet been developed for airborne viruses. A new sampling method for monitoring the personal exposure to bioaerosol particles has recently been developed and evaluated with bacteria and fungi. In this method, bacterial/fungal aerosol is aspirated and transported through a porous medium, which is submerged into a liquid layer. As the air is split into numerous bubbles, the particles are scavenged by these bubbles and effectively removed. The current feasibility study was initiated to evaluate the efficiency of the new personal sampler prototype ("bubbler") with airborne viable viruses. Two common viral strains, Influenza (stress-sensitive) and Vaccinia (robust), were aerosolized in the test chamber and collected by two identical "bubblers" that operated simultaneously for a duration of upto 5 min. A virus maintenance liquid, proven to be the optimum collecting environment for the test organisms, was used as a collection fluid. After sampling, the collecting fluid was analyzed and the viral recovery rate was determined. The overall recovery (affected not only by the sampling but also by the aerosolization and the aerosol transport) was 20% for Influenza virus and 89% for Vaccinia virus. The new sampling method was found feasible for the collection and enumeration of robust airborne viruses. ᭧

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A novel bioaerosol sampling technique, which utilizes the bubbling process in the collection fluid, has recently been developed and found feasible for a long-term personal sampling of airborne bacteria and fungal spores as it maintained high physical collection efficiency and high microbial recovery rate for robust and stresssensitive microorganisms. Further tests have shown that the new technique also has potential to collect viable airborne viruses, particularly when utilized for a short-term sampling of robust strains. As the short-term sampling has a limited application for assessing personal exposure in bioaerosol-contaminated environments, the present study was undertaken to investigate the feasibility of the "bubbler" for a long-term monitoring of viable airborne viruses. Liquid droplets containing Vaccinia virions (that simulate Variola, a causative agent of smallpox) were aerosolized with a Collison nebulizer into a 400-liter test chamber, from which the droplets were collected by three identical prototype personal samplers in the liquid medium during different time periods ranging from 1 to 6 hours. The viral content was measured in the collection fluid of the sampler and in the initial suspension of the nebulizer using the fluorescence-based method and by enumerating plaque-forming units per milliliter of the fluids. The relative recovery of viruses after the sampling act was determined. The results show that the "bubbling" technique has consistent collection efficiency over time and is capable of maintaining the viability of Vaccinia, for at least 6 hours, with a loss in recovery rate of about 10%. The data demonstrate a good potential of the new technique for measuring personal exposure to robust airborne viruses over a long period.