Effects of delayed microbial analysis of dental unit water line specimens

Monitoring microbial concentrations in water emitted from dental unit water lines (DUWL) is an important safety procedure. Improper handling of test water specimens could give incorrect results. Thus, the objective of this study was to measure the effects delayed culturing might have on DUWL specimens.

First, 100 mL water specimens were obtained from 10 different handpiece service lines within the School. All units had independent water systems, used DI (deionized water) water and were routinely cleaned using an alkaline peroxide based product. Two specimens of 10 mL were removed from the bottles and placed into individual sterile conical tubes. One set of tubes was processed immediately. 0.05 mL of sterile 1.0% (w/v) sodium thiosulfate solution was added to undiluted and diluted (1:10 and 1:100 with sterile DI water) specimens. After mixing, specimens were spiral plated onto duplicate R2A plates and incubated at 21degrees C for 7 days. Colonies were then counted and the cfu/mL of each original specimen determined. Another set of tubes was placed into a shipping envelope and mailed out to the School. Upon receipt, the tubes were processed as described above. The remaining 80 mL of water in the collection bottles were divided equally into new sterile tubes. One tube was left at 21 degrees C, while the other was placed into a 37 degrees C incubator. Aliquots were processed immediately and then after 1, 3 and 7 days. Next, 30 mL water specimens were obtained from 15 handpiece service lines in three outside clinics. All units had independent water systems, used DI water and were routinely cleaned with an alkaline peroxide-based product. Specimens were then divided equally into three sterile conical tubes. One of the tubes was transported (at 4 degrees C) to the laboratory and immediately processed as described. At the collection site, the second tube was placed into a padded envelope and mailed back to the School. The third tube was returned by overnight delivery using a Cool Pack type container. Upon receipt, all the tubes were processed as described previously.

The 10 handpiece waterline specimens processed immediately ranged from 0 to 1000 cfu/mL. Holding specimens at 21 degrees C produced radically higher bacterial counts (1540-866,000 cfu/mL) in water from 90% of the handpieces. Holding at 37 degrees C produced unacceptably high bacterial counts in only 50% of the handpiece specimens. Mailed specimens were cultured 5 days after collection and water of unacceptable quality water was noted in 70% of the specimens. In another experimental set, mailed specimens arrived after 72 hours and were an average of 20 degrees C. Express sent specimens came the next morning at an average temperature of 4.5 degrees C. Only one waterline specimen processed immediately contained more than 500 cfu/mL. In contrast, 80% of specimens returned by post at ambient temperature had unacceptably high bacterial counts (780-376,000 cfu/mL). Express sent specimens produced the same results as those processed immediately.