Evaluation of surface sampling techniques for collection of Bacillus spores on common drinking water pipe materials

Drinking water utilities may face biological contamination of the distribution system from a natural incident or deliberate contamination. Determining the extent of contamination or the efficacy of decontamination is a challenge, because it may require sampling of the wetted surfaces of distribution infrastructure. This study evaluated two sampling techniques that utilities might use to sample exhumed pipe sections. Polyvinyl chloride (PVC), cement-lined ductile iron, and ductile iron pipe coupons (3 cm x 14 cm) cut from new water main piping were conditioned for three months in dechlorinated Cincinnati, Ohio tap water. Coupons were spiked with Bacillus atrophaeus subsp. globigii, a surrogate for Bacillus anthracis. Brushing and scraping were used to recover the inoculated spores from the coupons. Mean recoveries for all materials ranged from 37 +/- 30% to 43 +/- 20% for brushing vs. 24 +/- 10% to 51 +/- 29% for scraping. On cement-lined pipe, brushing yielded a significantly different recovery than scraping. No differences were seen between brushing and scraping the PVC and iron pipe coupons. Mean brushing and scraping recoveries from PVC coupons were more variable than mean recoveries from cement-lined and iron coupons. Spore retention differed between pipe materials and the presence of established biofilms also had an impact. Conditioned PVC coupons (with established biofilms) had significantly lower spore retention (31 +/- 11%) than conditioned cement-lined coupons (61 +/- 14%) and conditioned iron coupons (71 +/- 8%).