Titrate by plaque assay in bacterial host to confirm that filtered media (phage lysate) contains around 1010 PFU/mL. Immediately before aerosolization, prepare 20 ml aliquots of 10-fold dilutions of the phage lysate. Use phage buffer for nebulization media. Keep nebulization at 4°C until use. Preparation of apparatus Autoclave the Collison nebulizer, BC 251 samplers and other sample extraction tools. Purge the test chamber by running the vacuum system for 30 minutes. Calibrate the air sampling pumps (flow rate of 4000 ± 40 mL/min) with DryCal DC-1. Connect sampling pumps to tubing. Place open-faced PTFE filter cassettes or NIOSH BC 251 samplers at sampling locations and connect by tubing to sampling pumps. Aerosol generation Take a 0.5 mL pre-sample of the liquid media and keep at 4°C. Φ6 Connect the Collison nebulizer to the bioaerosol inlet of the test chamber. Connect the Collison to the compressed air source. Transfer 15 mL of liquid media to the Collison reservoir. Φ6 Replace the Collison lid. Aerosolize at 20 psig for 20 minutes. Monitor the pressure Monitor the Collison reservoir for three round circles of impinging spray. the valve on the compressed air line as soon as aerosolization is complete. Remove the Collison and cover the bioaerosol inlet of the test chamber. Isolate the Collison from UVGI by moving it to adjacent room and closing the door. Take a 0.5 mL post-sample of the liquid media and keep at 4°C. Φ6 Pulsed germicidal irradiation Set the timer on the pulsed UVGI system according to desired exposure duration (0, 10, 20 or 30 seconds). Trigger the pulsed UVGI lamp as soon as the aerosol generation is complete and Collison is isolated. Bioaerosol sampling Run the air sampling pumps for 30 minutes as soon as the pulsed UVGI system automatically shuts off. Recover the filter cassettes or BC 251 samplers from the test chamber as soon as the sampling is complete. Bioaerosol sample extraction
Titrate by plaque assay in bacterial host to confirm that filtered media (phage lysate) contains around 1010 PFU/mL. Immediately before aerosolization, prepare 20 ml aliquots of 10-fold dilutions of the phage lysate. Use phage buffer for nebulization media. Keep nebulization at 4°C until use. Preparation of apparatus Autoclave the Collison nebulizer, BC 251 samplers and other sample extraction tools. Purge the test chamber by running the vacuum system for 30 minutes. Calibrate the air sampling pumps (flow rate of 4000 ± 40 mL/min) with DryCal DC-1. Connect sampling pumps to tubing. Place open-faced PTFE filter cassettes or NIOSH BC 251 samplers at sampling locations and connect by tubing to sampling pumps. Aerosol generation Take a 0.5 mL pre-sample of the Φ6 liquid media and keep at 4°C. Connect the Collison nebulizer to the bioaerosol inlet of the test chamber. Connect the Collison to the compressed air source. Transfer 15 mL of Φ6 liquid media to the Collison reservoir. Replace the Collison lid. Aerosolize at 20 psig for 20 minutes. Monitor the pressure Monitor the Collison reservoir for three round circles of impinging spray. the valve on the compressed air line as soon as aerosolization is complete. Remove the Collison and cover the bioaerosol inlet of the test chamber. Isolate the Collison from UVGI by moving it to adjacent room and closing the door. Take a 0.5 mL post-sample of the Φ6 liquid media and keep at 4°C. Pulsed germicidal irradiation Set the timer on the pulsed UVGI system according to desired exposure duration (0, 10, 20 or 30 seconds). Trigger the pulsed UVGI lamp as soon as the aerosol generation is complete and Collison is isolated. Bioaerosol sampling Run the air sampling pumps for 30 minutes as soon as the pulsed UVGI system automatically shuts off. Recover the filter cassettes or BC 251 samplers from the test chamber as soon as the sampling is complete. Bioaerosol sample extraction
UAST_1280128_SupplemenatlFile.docx