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Dose Determines if Soluble Copper is a Nutrient or an Antimicrobial for Legionella pneumophila in Premise Plumbing

The effect of copper on Legionella pneumophila in potable water plumbing systems is dependent on its dose and water chemistry. For instance, prior research demonstrated that the presence of aluminum hydroxide from anode rods in water heaters can bind copper and render high doses non-toxic. On this basis it was also hypothesized that iron hydroxide would have similar effects and that lower levels of copper may act as a nutrient encouraging Legionella growth. Here we conducted complementary experiments at bench, microcosm and pilot-scale to evaluate the effect of copper speciation and dose on L. pneumophila.

At bench-scale, the addition of 5 mg/L as Fe iron hydroxide to a solution with 1 mg/L copper decreased soluble copper from > 90% down to < 20% at pH 6.5-7. The reduction in soluble copper caused ~3-logs higher L. pneumophila culturability when iron was added with copper when compared to a condition with copper alone.

In a 9-month microcosm test using simulated glass water heaters with PEX pipe, a complete range of copper doses (0, 4, 30, 250 and 2000 g/L) were tested in triplicate. Over the first phase of research covered herein, the L. pneumophila levels were low at the four lowest doses of copper, and non-detectable at the highest dose. Moreover, total cell counts were highest at 250 g/L copper, lowest at 2000 g/L copper, and in between these extremes at the lower copper doses. This ongoing experiment will continue for months after this thesis was complete.

Pilot-scale experiments were conducted with anode rods removed from tank water heaters, to examine the effects of unprotected corrosion of the steel on iron release and Legionella pneumophila levels in systems with 1) added copper (WH-Cu), 2) copper and phosphate corrosion inhibitor (WH-Cu+PO4-3), 3) phosphate corrosion inhibitor alone (WH-PO4-3) and 4) a control with neither copper nor inhibitor (WH-Control). While there were slight differences in iron between the conditions, the iron concentration in the water of the tanks did not dramatically increase compared to when the powered anodes were still installed and reducing corrosion. Because the iron level was usually < 0.1 mg/L, the released iron dose was not high enough to reduce soluble copper. In fact, oddly, soluble copper increased by 37-183%, mostly likely because the installed anode rods were suppressing copper release.

Consequently, with the anode rods removed, the dose of 2000 g/L copper still had a strong antimicrobial effect. The levels of L. pneumophila increased in the order WH-Cu (2.6-logs CFU/L) < WH-PO4-3 (5.1-logs CFU/L) ≈ WH-Control (5.1-logs CFU/L). The addition of phosphate precipitated some of the added copper, such that the condition with copper and phosphate [WH-Cu+PO4-3 (4.2-logs CFU/L)] had L. pneumophila in between the condition with copper alone and that with no added copper.

When the copper dose in the pilot rig was reduced to 1000 g/L, Legionella increased somewhat in the system with added copper compared to the control, and L. pneumophila increased in the water heaters in the order WH-Cu (3.4-logs CFU/L) < WH-Cu+PO4-3 (4.3-logs CFU/L) < WH-PO4-3 (4.9-logs CFU/L) ≈ WH-Control (5.0-logs CFU/L). Overall, an antimicrobial effect of copper was maintained in the water heaters even after the anodes were removed and iron in the water increased slightly. If iron corrosion and release to water were much higher without the anode rods than observed in this study, it would be predicted that the higher iron would have reduced the copper antimicrobial effect. / Master of Science / The effect of copper on Legionella pneumophila in potable water plumbing systems depends on its dose and water chemistry. For instance, prior research demonstrated that the presence of aluminum hydroxide from anode rods in water heaters can bind copper and render high doses non-toxic. It was also hypothesized that iron hydroxide would have similar effects and that lower levels of copper may act as a nutrient encouraging Legionella growth. Here we conducted complementary experiments at bench, microcosm and pilot-scale to evaluate the effect of copper's chemistry and dose on L. pneumophila.

At bench-scale, the addition of a high level of iron to a solution with copper decreased the amount of copper available to Legionella from > 90% down to < 20% at pH 6.5-7. The reduction in bioavailable copper caused ~3-logs higher L. pneumophila when iron was added with copper when compared to a condition with copper alone.

In a 9-month microcosm test using simulated glass water heaters with PEX pipe, a complete range of copper doses (0, 4, 30, 250 and 2000 g/L) were tested. The L. pneumophila levels were low and there were no significant differences between the five doses in the beginning phases of the experiment reported herein. However, total cell counts were highest at 250 g/L copper, lowest at 2000 g/L copper, and in between these extremes at the lower copper doses. This suggests that 250 g/L may be optimal in encouraging overall microbial growth.

Pilot-scale experiments were conducted with anode rods removed from water heaters to examine the effects of unprotected corrosion of the steel tank on iron release and Legionella pneumophila levels. The four water heaters had added copper (WH-Cu), copper and phosphate corrosion inhibitor (WH-Cu+PO4-3), phosphate corrosion inhibitor alone (WH-PO4-3) and neither copper nor inhibitor (WH-Control). The iron concentration in the water of the tanks did not dramatically increase compared to when the powered anodes were used to reduce corrosion. Nevertheless, there were still slight differences in iron concentration between the replicate water heaters. When 2000 g/L copper was dosed, WH-Cu and WH-Control had statistically higher iron levels than WH-PO4-3, consistent with copper increasing corrosion of the unprotected tank. However, because the iron level was usually < 0.1 mg/L, the released iron dose was not high enough to reduce bioavailable copper. In fact, bioavailable copper went up when the anode rod was removed, most likely because the anode rods were protecting brass plumbing from corrosion.

With the anode rods removed a dose of 2000 g/L copper had a strong antimicrobial effect. The levels of L. pneumophila increased in the order WH-Cu < WH-PO4-3 ≈ WH-Control. The addition of phosphate reduced some of the bioavailable copper, such that the condition with copper and phosphate (WH-Cu+PO4-3) had L. pneumophila in between the condition with copper alone and that with no added copper.

When the copper dose in the water heaters was reduced to 1000 g/L, Legionella increased somewhat in the system with added copper compared to the control, and L. pneumophila increased in the order WH-Cu < WH-Cu+PO4-3 < WH-PO4-3 ≈ WH-Control. Overall, an antimicrobial effect of copper was maintained in water heaters even after the anodes were removed and even though iron in the water increased slightly. If iron corrosion and release to water was much higher than occurred in this study, it would be expected that the iron could have reduced the copper antimicrobial effect.

Identiferoai:union.ndltd.org:VTETD/oai:vtechworks.lib.vt.edu:10919/115603
Date17 August 2022
CreatorsFinkelstein, Rachel Briana
ContributorsEnvironmental Science and Engineering, Edwards, Marc A., Pruden, Amy, Falkinham, Joseph O.
PublisherVirginia Tech
Source SetsVirginia Tech Theses and Dissertation
LanguageEnglish
Detected LanguageEnglish
TypeThesis
FormatETD, application/pdf
RightsIn Copyright, http://rightsstatements.org/vocab/InC/1.0/

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