Jason Wylie

Influence of biofilms on iron and manganese deposition in drinking water distribution systems

Although health risk due to discoloured water is minimal, such water continues to be the source of one of the major complaints received by most water utilities in Australia. Elevated levels of iron (Fe) and/or manganese (Mn) in bulk water are associated with discoloured water incidents. The accumulation of these two elements in distribution systems is believed to be one of the main causes for such elevated levels. An investigation into the contribution of pipe wall biofilms towards Fe and Mn deposition, and discoloured water events is reported in this study. Eight laboratory-scale reactors were operated to test four different conditions in duplicate. Four reactors were exposed to low Fe (0.05 mg l−1) and Mn (0.02 mg l−1) concentrations and the remaining four were exposed to a higher (0.3 and 0.4 mg l−1 for Fe and Mn, respectively) concentration. Two of the four reactors which received low and high Fe and Mn concentrations were chlorinated (3.0 mg l−1 of chlorine). The biological activity (measured in terms of ATP) on the glass rings in these reactors was very low (∼1.5 ng cm−2 ring). Higher concentrations of Fe and Mn in bulk water and active biofilms resulted in increased deposition of Fe and Mn on the glass rings. Moreover, with an increase in biological activity, an increase in Fe and Mn deposition was observed. The observations in the laboratory-scale experiments were in line with the results of field observations that were carried out using biofilm monitors. The field data additionally demonstrated the effect of seasons, where increased biofilm activities observed on pipe wall biofilms during late summer and early autumn were found to be associated with increased deposition of Fe and Mn. In contrast, during the cooler months, biofilm activities were a magnitude lower and the deposited metal concentrations were also significantly less (ie a drop of 68% for Fe and 86% for Mn). Based on the laboratory-scale investigations, detachment of pipe wall biofilms due to cell death or flow dynamics could release the entrapped Fe and Mn into the bulk water, which could lead to a discoloured water event. Hence, managing biofilm growth on drinking water pipelines should be considered by water utilities to minimize accumulation of Fe and Mn in distribution networks.

Reduced Efficiency of Chlorine Disinfection of Naegleria fowleri in a Drinking Water Distribution Biofilm

Naegleria fowleri associated with biofilm and biological demand water (organic matter suspended in water that consumes disinfectants) sourced from operational drinking water distribution systems (DWDSs) had significantly increased resistance to chlorine disinfection. N. fowleri survived intermittent chlorine dosing of 0.6 mg/L for 7 days in a mixed biofilm from field and laboratory-cultured Escherichia coli strains. However, N. fowleri associated with an attached drinking water distribution biofilm survived more than 30 times (20 mg/L for 3 h) the recommended concentration of chlorine for drinking water. N. fowleri showed considerably more resistance to chlorine when associated with a real field biofilm compared to the mixed laboratory biofilm. This increased resistance is likely due to not only the consumption of disinfectants by the biofilm and the reduced disinfectant penetration into the biofilm but also the composition and microbial community of the biofilm itself. The increased diversity of the field biofilm community likely increased N. fowleris resistance to chlorine disinfection compared to that of the laboratory-cultured biofilm. Previous research has been conducted in only laboratory scale models of DWDSs and laboratory-cultured biofilms. To the best of our knowledge, this is the first study demonstrating how N. fowleri can persist in a field drinking water distribution biofilm despite chlorination.

Characterization of a Drinking Water Distribution Pipeline Terminally Colonized by Naegleria fowleri

Free-living amoebae, such as Naegleria fowleri, Acanthamoeba spp., and Vermamoeba spp., have been identified as organisms of concern due to their role as hosts for pathogenic bacteria and as agents of human disease. In particular, N. fowleri is known to cause the disease primary amoebic meningoencephalitis (PAM) and can be found in drinking water systems in many countries. Understanding the temporal dynamics in relation to environmental and biological factors is vital for developing management tools for mitigating the risks of PAM. Characterizing drinking water systems in Western Australia with a combination of physical, chemical and biological measurements over the course of a year showed a close association of N. fowleri with free chlorine and distance from treatment over the course of a year. This information can be used to help design optimal management strategies for the control of N. fowleri in drinking-water-distribution systems.

Comparison of biofilm ecology supporting growth of individual Naegleria species in a drinking water distribution system

Free-living amoebae (FLA) are common components of microbial communities in drinking water distribution systems (DWDS). FLA are of clinical importance both as pathogens and as reservoirs for bacterial pathogens, so identifying the conditions promoting amoebae colonisation of DWDSs is an important public health concern for water utilities. We used high-throughput amplicon sequencing to compare eukaryotic and bacterial communities associated with DWDS biofilms supporting distinct FLA species (Naegleria fowleri, N. lovaniensis or Vermamoeba sp.) at sites with similar physical/chemical conditions. Eukaryote and bacterial communities were characteristics of different FLA species presence, and biofilms supporting Naegleria growth had higher bacterial richness and higher abundance of Proteobacteria, Bacteroidetes (bacteria), Nematoda and Rotifera (eukaryota). The eukaryotic community in the biofilms had the greatest difference in relation to the presence of N. fowleri, while the bacterial community identified individual bacterial families associated with the presence of different Naegleria species. Our results demonstrate that ecogenomics data provide a powerful tool for studying the microbial and meiobiotal content of biofilms, and, in these samples can effectively discriminate biofilm communities supporting pathogenic N. fowleri. The identification of microbial species associated with N. fowleri could further be used in the management and control of N. fowleri in DWDS.

Competition between Naegleria fowleri and free living amoeba colonising laboratory scale and operational drinking water distribution systems

Free living amoebae (FLA), including pathogenic Naegleria fowleri, can colonize and grow within pipe wall biofilms of drinking water distribution systems (DWDSs). Studies on the interactions between various FLA species in biofilms are limited. Understanding the interaction between FLA and the broader biofilm ecology could help better predict DWDS susceptibility to N. fowleri colonization. The aim of this study was to determine if N. fowleri and other FLAs ( Naegleria, Vermamoeba, Willaertia, and Vahlkampfia spp.) cocolonize DWDS biofilm. FLAs commonly isolated from DWDSs ( N. fowleri, V. vermiformis, and N. lovaniensis) were introduced into laboratory-scale biomonitors to determine the impact of these amoebae on N. fowleris presence and viability. Over 18 months, a single viable amoebae ( N. fowleri, N. lovaniensis, or V. vermiformis) was detected in each biofilm sample, with the exception of N. lovaniensis and N. fowleri, which briefly cocolonized biofilm following their coinoculation. The analysis of biofilm and bulk water samples from operational DWDSs revealed a similar lack of cocolonization with a single FLA detected in 99% ( n = 242) of samples. Interestingly, various Naegleria spp. did colonize the same DWDS locations but at different times. This knowledge furthers the understanding of ecological factors which enable N. fowleri to colonize and survive within operational DWDSs and could aid water utilities to control its occurrence.

Effectiveness of Devices to Monitor Biofouling and Metals Deposition on Plumbing Materials Exposed to a Full-Scale Drinking Water Distribution System

A Modified Robbins Device (MRD) was installed in a full-scale water distribution system to investigate biofouling and metal depositions on concrete, high-density polyethylene (HDPE) and stainless steel surfaces. Bulk water monitoring and a KIWA monitor (with glass media) were used to offline monitor biofilm development on pipe wall surfaces. Results indicated that adenosine triphosphate (ATP) and metal concentrations on coupons increased with time. However, bacterial diversities decreased. There was a positive correlation between increase of ATP and metal deposition on pipe surfaces of stainless steel and HDPE and no correlation was observed on concrete and glass surfaces. The shared bacterial diversity between bulk water and MRD was less than 20% and the diversity shared between the MRD and KIWA monitor was only 10%. The bacterial diversity on biofilm of plumbing material of MRD however, did not show a significant difference suggesting a lack of influence from plumbing material during early stage of biofilm development.

Biological phosphorus and nitrogen removal in sequencing batch reactors: effects of cycle length, dissolved oxygen concentration and influent particulate matter

Removal of phosphorus (P) and nitrogen (N) from municipal wastewaters is required to mitigate eutrophication of receiving water bodies. While most treatment plants achieve good N removal using influent carbon (C), the use of influent C to facilitate enhanced biological phosphorus removal (EBPR) is poorly explored. A number of operational parameters can facilitate optimum use of influent C and this study investigated the effects of cycle length, dissolved oxygen (DO) concentration during aerobic period and influent solids on biological P and N removal in sequencing batch reactors (SRBs) using municipal wastewaters. Increasing cycle length from 3 to 6 h increased P removal efficiency, which was attributed to larger portion of N being removed via nitrite pathway and more biodegradable organic C becoming available for EBPR. Further increasing cycle length from 6 to 8 h decreased P removal efficiencies as the demand for biodegradable organic C for denitrification increased as a result of complete nitrification. Decreasing DO concentration in the aerobic period from 2 to 0.8 mg L(-1) increased P removal efficiency but decreased nitrification rates possibly due to oxygen limitation. Further, sedimented wastewater was proved to be a better influent stream than non-sedimented wastewater possibility due to the detrimental effect of particulate matter on biological nutrient removal.

Treatment of saline, acidic, metal-contaminated groundwater from the Western Australian Wheatbelt

Managing acidic, metal-containing saline ground and drainage waters in the Wheatbelt of Western Australia is an environmental and economic challenge. Sulfate-reducing fluidised bed bioreactors are shown to be technically capable of treating high salt, low pH, metal containing waters from the town of Narembeen in the Wheatbelt so as to reduce acidity and to remove most of the undesirable metal contaminants. The hydraulic residence time (HRT) limit for a stable process with groundwater from the region of Narembeen was >16 hours. The maximal rate of sulfate reduction in the laboratory system treating Narembeen groundwater was similar to rates observed in comparable applications of the process at other sites, ca. 3 g sulfate (L-reactor)(-1) day(-1). Salts that are relatively free of metal contaminants can be produced from water that has been treated by the sulfate-reducing fluidised bed bioreactor. It is unlikely that metal precipitates, captured from Wheatbelt waters by the process, would be of economic value. If sulfate-reducing fluidised bed reactors were considered technologically appropriate at larger scale, the decision to use them would be based on the necessity to take action, the comparative effectiveness of competing technologies, and the relative costs of competing technologies.

Two-Stage Airlift Bioreactor System for Efficient Iron Oxidation and Jarosite Precipitation

Continuous high-rate iron oxidation and removal of jarosite precipitates from solution at low pH and ambient temperature and pressure was successfully demonstrated. The bio-catalysed iron oxidation and jarosite precipitation is promising as a unit process for a variety of hydrometallurgical process flow sheets, where it allows for iron removal from ferrous solutions without the requirement for chemical addition and with negligible base metal co-precipitation losses. The process demonstrated performance that could be used in a large scale industry unit. A two-stage airlift bioreactor (ALBR) system comprised of two ALBRs, each with its own settler, was operated for iron oxidation and precipitation at room temperature with a mixed culture of mesophilic iron oxidisers. The two-stage reactor design allowed for optimization of overall reactor kinetics by facilitating the growth of low (430 mV vs Ag/AgCl) and high (517 mV) redox potential iron oxidizers in the respective reactors. The influent (pH 1.5) contained (g L-1) 15 Fe2+, 1.5 Cu, 1.5 Ni, nutrients and trace elements. The hydraulic retention time (HRT) was decreased stepwise to evaluate process performance. With the lowest HRTs (8 h in ALBR1 and 10 h in ALBR2), the overall iron oxidation and precipitation rates of the two-stage system were 0.75 ± 0.02 g L-1 h-1 and 0.15 ± 0.01 g L-1 h-1, respectively and overall iron oxidation and precipitation efficiencies of 94 ± 3% and 18 ± 1 %, respectively. The percent of influent Fe, S, Cu and Ni removed as precipitates from settlers were 30.9%, 16.7%, 1.1% and 0.2%, respectively. The precipitates were predominately comprised of (>95%) jarosite with potassium jarosite being the dominant form, followed by hydronium, ammonium and sodium jarosites. In conclusion, the two-stage ALBR system allowed efficient iron oxidation and precipitation of the oxidised iron as well settling jarosite with only minor loss of Cu and Ni via co-precipitation.