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Previous studies indicated evidence of opportunistic pathogens in samples obtained during missions to the International Space Station (ISS). This study utilized TaqMan quantitative PCR to determine specific gene abundance in potable and non-potable ISS waters. Probe and primer sets specific to the small subunit rRNA genes were designed and used to elucidate overall bacterial rRNA gene numbers. In addition, primer-probe sets specific for Burkholderia cepacia and Stenotrophomonas maltophilia were optimized and genes of these two opportunistic pathogens quantified in the pre- and post-flight drinking water as well as coolant waters. This Q-PCR approach supports findings of previous culture-based studies however; the culture based studies may have underestimated the microbial burden of ISS drinking water.

Certain Eubacteria enter a viable but nonculturable (VBNC) state upon encountering unfavorable environmental conditions. VBNC cells do not divide on conventional media yet remain viable and in some cases retain virulence. Here, we describe the VBNC state of two opportunistic pathogens previously isolated from ISS potable waters, Burkholderia cepacia and Stenotrophomonas maltophilia. Artificially inoculated microcosms were exposed to the biocidal agents copper (CuSO4) and iodine (I2) in an attempt to induce nonculturablility. Viability was assessed via fluorescent microscopy (direct viable count assay coupled with BacLight™ staining) and metabolic activity was monitored by quantifying both intracellular ATP and transcribed rRNA (reverse transcriptase quantitative PCR). Culturablility was lost in both B. cepacia and S. maltophilia within two days of exposure to copper or high concentrations of iodine (6 or 8 ppm).