Short communicationGeneration of avian influenza virus (AIV) contaminated fecal fine particulate matter (PM2.5): Genome and infectivity detection and calculation of immission
Introduction
Some of the most highly contagious viruses of human and veterinary concern are airborne pathogens, e.g. human and avian influenza virus (AIV) and foot-and-mouth disease virus (FMDV). The highly pathogenic avian influenza viruses (HPAI) have been a threat to the poultry industry for many years. However, when the Asian H5N1 subtype emerged, a trans-boundary spread became evident and a new dimension of zoonotic danger and fear for evolution of a pandemic virus was raised (Belshe, 2005, Peiris et al., 2007, World Health Organization, 2007). After the first localized Asian H5N1 epidemic in Germany, the virus was spread rapidly and caused many small epidemics in wild birds. In 2007, the virus also was detected on duck raising farms (Rinder et al., 2007, Starick et al., 2007, Weber et al., 2007, Harder et al., 2009). The routes of transmission of Asian H5N1 and other HPAI subtypes among susceptible mammals and birds is still a matter of debate. Air borne transmission of infection cannot be excluded and might be an effective way of infection transmission when virus is inhaled directly in the deeper respiratory tract (Shinya et al., 2006, Tellier, 2006, Brankston et al., 2007, Nicholls et al., 2007). Fine particulate matter from AIV contaminated fecal dust can play an important role in the transmission and onset of infection since particles as small as PM2.5 (particle size <2.5 μm) can directly invade the lower part of the respiratory tract (DFG, 2008). Rapid and direct transport of AIV into the lungs will be in favor for onset of infection (Shinya et al., 2006, Van Riel et al., 2006). The indoor exposure to aerosolized HPAI-AIV is equally given for humans working in poultry stables and susceptible birds. As workers can prevent virus inhalation by using appropriate masks birds are most endangered by aerosolized AIV within the flocks and possibly in nearby holdings.
Recently the stability and transmission of aerosolized Newcastle Disease Virus has been tested under practical conditions with chickens (Li et al., 2009). The results indicated that virus shed from infected animals readily aerosolized and airborne transmission to susceptible chickens was very efficient. We therefore wanted to know, (i) whether AIV in contaminated fecal dust can be found in the PM2.5 fraction and, (ii) if this is the case, how long viable virus is present under environmental conditions.
Section snippets
AIV stock solution
The low pathogenic avian influenza virus (LPAI) subtype H10N7 has been isolated and multiplied in 10-day-old embryonated chicken eggs as described (Rinder et al., 2007). Allantoic fluid was collected at the time of embryonic death, clarified by centrifugation at 800 × g, the supernatants were pooled and the virus was pelleted by ultra centrifugation in a Beckman Coulter Optima L-90K ultra centrifuge. The pellets were pooled, re-suspended in phosphate buffer and stored at −70 °C. The concentrated
Generation of filter samples coated with AIV contaminated fecal PM2.5
Particle size distributions of resulting aerosols showed – with high conformity – maxima of particle numbers at 0.4–0.6 μm particle diameter (Fig. 2a) and maxima of particle volumes (or masses) at 2–4 μm particle diameter (Fig. 2b). More than 90% of the generated particles had sizes <1.5 μm, and therefore represent particulate matter with a size range less than 2.5 μm (PM2.5) which can directly enter the alveolar part of the lungs (DFG, 2008).
The aerosol generator produced very high concentrations
Discussion
The most common methods for viral aerosol sampling are liquid impactors, solid impactors combined with petri dishes and different filter types such as cellulose, polycarbonate, gelatine and PTFE (Verreault et al., 2008). So far, influenza virus in bioaerosol has most frequently been collected by liquid impactors. Recently, Pyankov et al. (2007) also performed the collection of airborne AIV (LPAI H11N9) in an open channel type aerosol chamber using liquid impactors in combination with real-time
Conclusion
As poultry farms are known to emit fine particulate matter (PM2.5 and PM10) which is composed of the abrasions from feces and litter material, they are a potential risk for airborne neighborhood infection. According to the results from a dispersion calculation for a 120,000 animal broiler farm, assuming a worst case scenario for an adult person, she/he could inhale up to 104 infectious AIV per day living in a nearby settlement. The 50% human infective dose by aerosol inoculation of influenza
Acknowledgements
This project was funded by the Bavarian State Ministry of the Environment, Public Health and Consumer Protection.
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