1. Effectiveness of Spore Trap Only Sampling Richard Billups Laboratory Director Air Allergen & Mold Testing Atlanta, Ga.

2. SPORE TRAPS  main tool used by most investigators in the field to diagnose indoor air quality complaints  too much reliance upon spore trap testing only? Air Allergen & Mold Testing

3. Basis of Experiment  compare use of spore traps alone and then in conjunction with carpet dust culture from homes and businesses with compaints of allergies or asthma Air Allergen & Mold Testing

4. Basis of Experiment  spore traps give information for the current condition of the site  spore traps DO NOT provide species ID of Aspergillus / Penicillium  carpet dust provides the history of the site  carpet dust culture can provide species ID of Aspergillus, Penicillium Air Allergen & Mold Testing

5. Parameters Spore Trap  > 1,000 Total Spores  > 500 spores Aspergillus / Penicillium  Combination of any high water activity spores Dust Culture  Bulk dust from vacuum of carpet  < 10,000 CFU / gram = Low  10,000 -50,000 CFU / gram = Moderate  50,000 – 100,000 CFU / gram = High  > 100,000 CFU / gram = Very High Air Allergen & Mold Testing11

6. Parameters  Samples collected during June, July, August in the Atlanta, Ga area, 2010  Allergenco D spore traps  Dust samples collected with a portable Dyson  Dust collection units from Indoor Biotechnologies  Dust cultured in dilutions on MEA 2%, DG18, Cellulose agars Air Allergen & Mold Testing11

7. Parameters  Identified dust culture organisms to species where possible  Used Group I list from ERMI as indicators of high water activity  Added additional organisms known to be associated with high water activity  Did not subtract out phyloplane organisms Air Allergen & Mold Testing

8. Organisms of Interest ERMI Group I Aspergillus flavus Aspergillus fumigatus Aspergillus niger Aspergillus ochraceus Aspergillus penicilloides Aureobasidium pullulans Aspergillus restrictus Aspergillus sclerotiorum Aspergillus sydowii Aspergillus uniguis Aspergillus versicolor Chaetomium globosum Cladosporium sphaerospermum Eurotium amstelodami Additional Acremonium sp Aspergillus usuts Chaetomium sp. Eurotium herbariorum Penicillium chrysogenum Rhizopus sp Rhodotorula sp Trichoderma harzianum Trichoderma sp Ulocladium sp Air Allergen & Mold Testing Paecilomyces variotii Penicillium brevicompactum Penicillium corylophilum Penicillium crustosum Penicillium purpurogenum Penicillium spinulosum Penicillium variabile Scopulariopsis brevicaulis Scopulariopsis chartarum Stachybotrys chartarum Trichoderma viride Wallemia sebi

9. RESULTS  spore traps with > 1,000 Aspergillus/ Penicillium = 5  spore trap with > 500 Aspergillus / Penicillium = 1 23% Median Total Spores = 784 spores/ m 3 Median with total < 1,000 = 471 spores / m 3 Air Allergen & Mold Testing

10. Results  carpet dust < 10,000 CFU /g = 6 26% 10,000 50,000 CFU /g = 7 30% 50,000- 100,000 CFU/g = 3 13% > 100,000 CFU/g = 7 30% Combination of > 10,000 and above = 73.9 % Samples with no resolution = 3 11.5% Median = 38,149 CFU / g Air Allergen & Mold Testing

11. Results  Organisms Recovered from dust culture Penicillium sp 16 Aspergillus sydowii 5 Aspergillus versicolor 16 Cladosporium cladosporioides 4 Cladosporium sp 15 Aspergillus ochraceus 4 Curvularia sp 13 Aspergillus ustus 4 Rhodotorula sp 12 Bipolaris sp 3 Acremonium sp 11 Phoma sp 3 Eurotium amstelodami 11 Aspergillus flavus 3 Aspergillus niger 10 Pithomyces sp 3 Stachybotrys chartarum 9 Trichoderma harzianum 2 Epicoccum nigrum 8 Aureobasidium pullulans 2 Rhizopus sp 8 Penicillium chrysogenum 2 Fusarium sp 7 Aspergillus fumigatus 2 Mucor sp 7 Yeast 6 Air Allergen & Mold Testing

16. What Did We Learn  Spore trap action level of 1,000 not reliable if Aspergillus, Penicillium or high water activity organisms absent  Spore trap only sampling not reliable as a method to determine fungal contamination of the indoor air (fall, winter?) Air Allergen & Mold Testing

17. What Did We Learn  analyzing carpet dust provides an increased ability to solve indoor air quality issues due to fungal contaminants  identification of organisms to the species level for Aspergillus, Penicillium helpful in assessing possible contamination  number of CFU’s may not be as important as the type of organisms recovered, especially if the organisms indicate high water activity  Aspergillus versicolor dominated the number of times recovered as a species 66.7% Air Allergen & Mold Testing

18. What Did We Learn  38,149 CFU/g median -- what number should provide the best information for action ?  Dust carpet culture should utilize multiple plate agar methodology to include: MEA 2% DG18 Cellulose  Other agars as needed, depending on the site and information provided by the investigator (ie PDA, SSA, etc) Air Allergen & Mold Testing

19. What Next  Analyzing culture carpet dust direct, dilution, bulk, refined  More culture from HVAC supply  Physician analysis of blood bio markers  Building materials, especially carpet  Vacuum cleaner comparisons and use Air Allergen & Mold Testing

20. Entire paper will be available on our web site by June 1 www.mold-testing-lab.com Will also be discussed on our blog Can reach me by Email rbillups@airallergen.com Air Allergen & Mold Testing