Airborne bacterial spore counts by terbium-enhanced luminescence detection: Pitfalls and real values

Bacterial spore determination by terbium(III)-dipicolinate luminescence has been reported by several investigators. We collected spore samples with a cyclone and extracted dipicolinic acid (DPA) in-line with hot aqueous dodecylamine, added Tb(III) in a continuous-flow system and detected the Tb(Ill)DPA with a gated liquid core waveguide fluorescence detector with a flashlamp excitation source. The absolute limit of detection (LOD) for the system was equivalent to 540 B. subtilis spores (for a 1.8 m(3) sample volume (t = 2 h, Q = 15 L/min), concentration LOD is 0.3 spores/L air). Extant literature suggests that, from office to home settings, viable spore concentrations range from 0.1 to 10 spores/L; however, these data have never been validated, Previously reported semi-automated instrumentation had an LOD of 50 spores/L. The present system was tested at five different location settings in Lubbock, Texas. The apparent bacterial spore concentrations ranged from 9 to 700 spores/L and only occasionally exhibited the same trend as the simultaneously monitored total optical particle counts in the >= 0.5 mu m size fraction. However, because the apparent spore counts sometimes were very large relative to the 0.5+ mu m size particle counts, we investigated potential positive interferences. We show that aromatic acids are very likely large interferents. This interference typically constitutes similar to 70% of the signal and can be as high as 95%. It can be completely removed by prewashing the particles.