Assessing residential activity in a home plumbing system simulator: monitoring the occurrence and relationship of major opportunistic pathogens and phagocytic amoebas.

Opportunistic premise plumbing pathogens (OPPPs) have been detected in buildings' plumbing systems causing waterborne disease outbreaks in the United States. In this study, we monitored the occurrence of OPPPs along with free-living amoeba (FLA) and investigated the effects of residential activities in a simulated home plumbing system (HPS). Water samples were collected from various locations in the HPS and analyzed for three major OPPPs: Legionella pneumophila, nontuberculous mycobacterial species (e.g., Mycobacterium avium, M. intracellulare, and M. abscessus), and Pseudomonas aeruginosa along with two groups of amoebas (Acanthamoeba and Vermamoeba vermiformis). A metagenomic approach was also used to further characterize the microbial communities. Results show that the microbial community is highly diverse with evidence of spatial and temporal structuring influenced by environmental conditions. L. pneumophila was the most prevalent pathogen (86% of samples), followed by M. intracellulare (66%) and P. aeruginosa (21%). Interestingly, M. avium and M. abscessus were not detected in any samples. The data revealed a relatively low prevalence of Acanthamoeba spp. (4%), while V. vermiformis was widely detected (81%) across all the sampling locations within the HPS. Locations with a high concentration of L. pneumophila and M. intracellulare coincided with the highest detection of V. vermiformis, suggesting the potential growth of both populations within FLA and additional protection in drinking water. After a period of stagnation lasting at least 2-weeks, the concentrations of OPPPs and amoeba immediately increased and then decreased gradually back to the baseline. Furthermore, monitoring the microbial population after drainage of the hot water tank and partial drainage of the entire HPS demonstrated no significant mitigation of the selected OPPPs. This study demonstrates that these organisms can adjust to their environment during such events and may survive in biofilms and/or grow within FLA, protecting them from stressors in the supplied water.

Sediment Disturbance Negatively Impacts Methanogen Abundance but Has Variable Effects on Total Methane Emissions.

Methane emissions from aquatic ecosystems are increasingly recognized as substantial, yet variable, contributions to global greenhouse gas emissions. This is in part due to the challenge of modeling biologic parameters that affect methane emissions from a wide range of sediments. For example, the impacts of fish bioturbation on methane emissions in the literature have been shown to result in a gradient of reduced to enhanced emissions from sediments. However, it is likely that variation in experimental fish density, and consequently the frequency of bioturbation by fish, impacts this outcome. To explore how the frequency of disturbance impacts the levels of methane emissions in our previous work we quantified greenhouse gas emissions in sediment microcosms treated with various frequencies of mechanical disturbance, analogous to different levels of activity in benthic feeding fish. Greenhouse gas emissions were largely driven by methane ebullition and were highest for the intermediate disturbance frequency (disturbance every 7 days). The lowest emissions were for the highest frequency treatment (3 days). This work investigated the corresponding impacts of disturbance treatments on the microbial communities associated with producing methane. In terms of total microbial community structure, no statistical difference was observed in the total community structure of any disturbance treatment (0, 3, 7, and 14 days) or sediment depth (1 and 3 cm) measured. Looking specifically at methanogenic Archaea however, a shift toward greater relative abundance of a putatively oxygen-tolerant methanogenic phylotype (ca. Methanothrix paradoxum) was observed for the highest frequency treatments and at depths impacted by disturbance (1 cm). Notably, quantitative analysis of ca. Methanothrix paradoxum demonstrated no change in abundance, suggesting disturbance negatively and preferentially impacted other methanogen populations, likely through oxygen exposure. This was further supported by a linear decrease in quantitative abundance of methanogens (assessed by qPCR of the mcrA gene), with increased disturbance frequency in bioturbated sediments (1 cm) as opposed to those below the zone of bioturbation (3 cm). However, total methane emissions were not simply a function of methanogen populations and were likely impacted by the residence time of methane in the lower frequency disturbance treatments. Low frequency mechanical disruption results in lower methane ebullition compared to higher frequency treatments, which in turn resulted in reduced overall methane release, likely through enhanced methanotrophic activities, though this could not be identified in this work. Overall, this work contributes to understanding how animal behavior may impact variation in greenhouse gas emissions and provides insight into how frequency of disturbance may impact emissions.

Bioturbation frequency alters methane emissions from reservoir sediments.

Inland aquatic systems are major global contributors to the atmospheric carbon budget through greenhouse gas (GHG) emissions, although the amount and form of carbon released varies widely across and within systems. Bioturbation of aquatic sediments can impact biogeochemical conditions and physically release sediment-bound bubbles containing GHGs, but variation in the frequency of such disturbance may modify the rate and composition of resulting GHG emissions. We hypothesized that an intermediate bioturbation frequency would result in the greatest methane (CH4) releases due to mechanical release of trapped bubbles, while frequent disturbance would result in greater diffusive carbon dioxide (CO2) releases relative to CH4, due to increased aeration of the sediment. We tested this bioturbation frequency hypothesis using laboratory mesocosms containing homogenized reservoir sediment. We used mechanical disturbance to simulate bioturbation at 3, 7, 14, or 21-day intervals; a control treatment was undisturbed for the duration of the experiment. We measured GHG emission (ebullition and diffusion) rates. An intermediate frequency of disturbance (7 days) produced the highest total GHG emission rate, while the most frequent disturbance interval (3 days) and least frequent interval (0 days) reduced overall GHG emissions relative to weekly disturbance by 24% and 15%, respectively. These patterns were primarily driven by differences in CH4 ebullition. Contrary to our hypothesis, there was no relationship between disturbance frequency and diffusive CO2 emissions. For all disturbance treatments, the majority of ebullition occurred during disturbance events, suggesting mechanical release of entrapped bubbles is an important emission mechanism. The frequency of disturbance has variable effects on GHG emissions and may explain conflicting results in prior studies of bioturbation. Our study provides insight into bioturbation as a driver of within-system variation in GHG emissions and highlights that variable bioturbation frequency results in non-linear responses in CH4 emissions, a globally important GHG, from reservoir sediments.