Indoor Air Quality Testing with an Andersen N6 Microbial Impactor Kit John Kanzia Environmental Quality Manager Chicago Zoological Society

Sampling Technique All samples taken with a single-stage Andersen N6 Microbial Impactor at 28.3 L/min flow Agar: Dichloran Glycerol, 18% Chloramphenicol (DG18) – selective for certain molds and fungi, including Aspergillus (mold), Penicillium (fungus), Candida (yeast), and Saccharomyces (yeast) Sample time = 3 minutes Plates incubated at 250 C for 72 hours Plates counted at 24, 48 and 72 hours Plates digitally photographed after 72 hours of incubation Several hundred species of Aspergillus and Penicillium, so initial focus was placed on enumeration rather than identification

Andersen N6 Microbial Impactor Kit Comprised of 4 components Vacuum pump, hose, impactor (inlet cone, jet stage, base plate), and nutrient agar plate Air enters through the inlet cone, and velocity is increased as air passes through the jet stage Small particles avoid plate impact and are filtered from the vacuum pump exhaust Larger particles (airborne mold and fungal spores) impact the plate and are cultured for enumeration during incubation

Andersen N6 Microbial Impactor Kit

Andersen N-6 Microbial Impactor Vacuum Pump

Examples of Yeasts and Molds That Will Grow on DG18 Agar Examples of Aspergillus and Pennicillium Photo by: Dr. David Midgley Cultures: Dr. David Midgley University of Sydney, Australia. Example of Candida (yeast) Thaimedicalnews.com

Site 1 Before Renovation 47 spores/m3 After Renovation 0 spores/m3

Site 2 Before Renovation 482 spores/m3 After Renovation 0 spores/m3

Site 3 Before Renovation 447 spores/m3 After Renovation 0 spores/m3

Site 4 Before Renovation 718 spores/m3 After Renovation 0 spores/m3

Site 5 Before Renovation 24 spores/m3 After Renovation 0 spores/m3

Site 6 Before Renovation 82 spores/m3 After Renovation 0 spores/m3

Site 7 Before Renovation 635 spores/m3 After Renovation 12 spores/m3

Site 8 Before Renovation 71 spores/m3 After Renovation 12 spores/m3

Site 9 Before Renovation 612 spores/m3 After Renovation 0 spores/m3

Outside Comparison Before Renovation 94 spores/m3 After Renovation

Data Table Exhibit/Area 04/21/08 Before Renovation Spores/m3 09/28/10 After Renovation Site 1 47 Site 2 482 Site 3 447 Site 4 718 Site 5 24 Site 6 82 Site 7 635 12 Site 8 71 Site 9 612 Outside Comparison 94 35

Renovation Details Related to Air Quality All live plants were removed and replaced with synthetic plants All soil was removed Planter boxes were demolished or repurposed Ductwork into the stadium was sealed from soil Fans added over north and south holding pools to improve air circulation at the air/water interface Building HVAC system was upgraded Stadium was thoroughly cleaned and repainted

Before Renovation

After Renovation

Next Steps Continue to perform seasonal air quality testing at location and other areas where vets have seen Aspergillosis cases Proactively seek out and sample exhibits with environmental conditions that favor the growth of Aspergillus (live plants, soil, high humidity, poor air circulation) Prepare DG18 agar in-house to reduce costs and wait time for prepared plates Analyze tape samples from plates to learn to identify commonly found molds and fungi

Sources 1. Dykstra, M., M. Loomis, K. Reninger, D. Zombeck and T. Faucette (1997). A Comparison of Sampling Methods for Airborne Fungal Spores During an Outbreak of Aspergillosis in the Forest Aviary of the North Carolina Zoological Park. Journal of Zoo and Wildlife Medicine 28 (4), 454-463. 2. Wilson, S.C., D.C. Strauss (2002). The Presence of Fungi Associated with Sick Building Syndrome in North American Zoological Institutions. Journal of Zoo and Wildlife Medicine 33 (4), 322-327. 3. J.D. Cooly, W.C. Wong, C.A. Jumper, D.C. Strauss (1998). Correlation Between the Prevalence of Certain Fungi and Sick Building Syndrome. Occupational Environmental Medicine 55, 579-584.