The free-living amoeboflagellate Naegleria fowleri is the causative agent of primary amoebic meningoencephalitis (PAM), a rapidly fatal disease of the central nervous system. In the United States, the disease is generally acquired while swimming and diving in freshwater lakes and ponds. In addition to swimming, exposure to N. fowleri and the associated disease can occur by total submersion in bathwater or small backyard wading pools. In the present study, swipe samples and residual pipe water from homes in Arizona were examined for N. fowleri by nested PCR due to the death of two previously healthy children from PAM. Since neither child had a history of swimming in a freshwater lake or pond prior to the onset of disease symptoms, the domestic water supply was the suspected source of infection. Of 19 samples collected from bathroom and kitchen pipes and sink traps, 17 samples were positive for N. fowleri by PCR. A sample from a Micro-Wynd II filter was obtained by passing water from bathtubs through the filter. Organisms attached to the filter also tested positive by PCR. The two samples that tested negative for N. fowleri were one that was obtained from a kitchen sink trap and a swipe sample from the garbage disposal of one home.

Trichloroethene (TCE) and perchlorate (ClO4 –) are cocontaminants at multiple Superfund sites. Fe0 is often used during TCE bioremediation with Dehalococcoides mccartyi to establish anoxic conditions in the aquifer. However, the synergy between Fe0 abiotic reactions and microbiological TCE and ClO4 – reductions is poorly understood and seldom addressed in the literature. Here, we investigated the effects of Fe0 and its oxidation product, Fe2 +, at field-relevant concentrations in promoting microbial TCE and ClO4 – reductions. Using semibatch microcosms with a Superfund site soil and groundwater, we showed that the high Fe0 concentration (16.5 g L–1) expected during Fe0 in situ injection mostly yielded TCE abiotic reduction to ethene/ethane. However, such concentrations obscured dechlorination by D. mccartyi, impeded ClO4 – reduction, and enhanced SO4 2– reduction and methanogenesis. Fe2 + at 0.25 g L–1 substantially delayed conversion of TCE to ethene when compared to no-Fe controls. A low concentration of aged-Fe0 synergistically promoted microbiological TCE dechlorination to ethene while achieving complete ClO4 – reduction. Collectively, these results illustrate scenarios relevant at or downstream of Fe0 injection zones when Fe0 is used to facilitate microbial dechlorination. Results also underscore the potential detrimental effects of Fe0 and bioaugmentation cultures coinjection for in situ treatment of chlorinated ethenes and ClO4 –.

Secondary substrate utilization of methylene chloride was analyzed by using Pseudomonas sp. strain LP. Both batch and continuously fed reactors demonstrated that this strain was capable of simultaneously consuming two substrates at different concentrations: the primary substrate at the higher concentration (milligrams per liter) and the secondary substrate at the lower concentration (micrograms per liter). The rate of methylene chloride utilization at trace concentrations was greater in the presence of the primary substrate, acetate, than without it. However, when the substrate roles were changed, the acetate secondary substrate utilization rate was less when methylene chloride was present. Thus, substrate interactions are important in the kinetics of secondary substrate utilization. Pseiudomonas sp. strain LP showed a preference toward degrading methylene chloride over acetate, whether it was the primary or secondary substrate, providing it was below an inhibitory concentration of ca. 10 mg/liter. Methylene chloride (dichloromethane) is designated as a priority pollutant by the U.S. Environmental Protection Agency (20). It is widely used for cleaning, fumigation, refrigeration, paint removal, analytical applications, and as an aerosol propellant (23). Treated municipal wastewaters have been recorded to contain as much as 24 [.g of methyl

Fe0 is a powerful chemical reductant with applications for remediation of chlorinated solvents, including tetrachloroethene and trichloroethene. Its utilization efficiency at contaminated sites is limited because most of the electrons from Fe0 are channeled to the reduction of water to H2 rather than to the reduction of the contaminants. Coupling Fe0 with H2-utilizing organohalide-respiring bacteria (i.e., Dehalococcoides mccartyi) could enhance trichloroethene conversion to ethene while maximizing Fe0 utilization efficiency. Columns packed with aquifer materials have been used to assess the efficacy of a treatment combining in space and time Fe0 and aD. mccartyi-containing culture (bioaugmentation). To date, most column studies documented only partial conversion of the solvents to chlorinated byproducts, calling into question the feasibility of Fe0 to promote complete microbial reductive dechlorination. In this study, we decoupled the application of Fe0 in space and time from the addition of organic substrates andD. mccartyi-containing cultures. We used a column containing soil and Fe0 (at 15 g L–1 in porewater) and fed it with groundwater as a proxy for an upstream Fe0 injection zone dominated by abiotic reactions and biostimulated/bioaugmented soil columns (Bio-columns) as proxies for downstream microbiological zones. Results showed that Bio-columns receiving reduced groundwater from the Fe0-column supported microbial reductive dechlorination, yielding up to 98% trichloroethene conversion to ethene. The microbial community in the Bio-columns established with Fe0-reduced groundwater also sustained trichloroethene reduction to ethene (up to 100%) when challenged with aerobic groundwater. This study supports a conceptual model where decoupling the application of Fe0 and biostimulation/bioaugmentation in space and/or time could augment microbial trichloroethene reductive dechlorination, particularly under oxic conditions.

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