Assessing the validity and sensitivity of microbial processes within a hydrodynamic model.

Eutrophication due to excess anthropogenic nutrients in waterways is a significant issue worldwide. The pressure-stressor-response of a waterway to excessive nutrient loading is reliant on numerous physical and biological factors, including hydrodynamics and microbial processing. While substantial progress has been made towards simulating these mechanisms there are limited multi-disciplinary studies that relate the physical hydrodynamics of a site with the ecological response from linked laboratory and field studies. This paper presents the development of a coupled hydrodynamic and aquatic ecosystem response model, expanded to include an integrated microbial loop, that allows the explicit representation of heterotrophic bacteria growth and dissolved organic nutrient mineralisation. A unique long-term water quality dataset at an estuary in south-eastern Australia was used to validate and assess the model's sensitivity to complex biophysical processes driving the observed water quality variability. Results indicate that explicit time-varying bacterial mineralisation rates provide a substantially improved understanding of the broader aquatic ecosystem response than assigned fixed bulk rate parameter values, which are typically derived from non-local literature. Implementation of a microbial loop at the study site indicated that the model is sensitive to the boundary conditions, in particular catchment loads, with both net transport rates and the net growth rates of heterotrophic bacteria demonstrating different responses. Under average flow conditions, a smaller net transport and reduced nutrient availability has a pronounced effect of lowering net growth rates through the applied limitation factors. During high flow conditions, freshwater inflows increased net transport and nutrient loads, which resulted in higher net growth rates. Further, temporal variability in water temperature had a compounding effect on the model's response sensitivity. This approach has broader application in other riverine systems subject to eutrophication, and in interrogating linkages in hydrodynamic and microbial mediated processes (e.g., productivity). Future studies are recommended to better understand the sensitivity of aquatic ecosystem response models to microbial net growth rate kinetics at different temperatures and from top-down predation (e.g., zooplankton grazing).

Implications of bacterial mineralisation in aquatic ecosystem response models.

Widespread wastewater pollution is a major barrier to the sustainable management of freshwater and coastal marine ecosystems worldwide. Integrated multi-disciplinary studies are necessary to improve waterway management and protect ecosystem integrity. This study used the Generalised Likelihood Uncertainty Estimation (GLUE) methodology to link microbial community ecotoxicology laboratory data to a mechanistic aquatic ecosystem response model. The generic model provided good predictive skill for major water quality constituents, including heterotrophic bacteria dynamics (r2 = 0.91). The model was validated against observed data across a gradient of effluent concentrations from community whole effluent toxicity (WET) laboratory tests. GLUE analysis revealed that a combined likelihood measure increased confidence in the predictive capability of the model. This study highlights the importance of calibrating aquatic ecosystem response models with net growth rates (i.e., sum of the growth minus loss rate parameter terms) of biological functional groups. The final calibrated net growth rate value of heterotrophic bacteria determined using the GLUE analysis was selected to be 0.58, which was significantly greater than the average literature value of -0.15. This finding demonstrated that use of literature parameter values without a good understanding of the represented processes could create misleading outputs and result in unsatisfactory conclusions. Further, fixed bulk mineralisation rate literature values are typically higher than realistically required in aquatic ecosystem response models. This indicates that explicitly including bacterial mineralisation is crucial to represent microbial ecosystem functioning more accurately. Our study suggests that improved data collection and modelling efforts in real-world management applications are needed to better address nutrients released into the natural environment. Future studies should aim to better understand the sensitivity of aquatic ecosystem response models to bacterial mineralisation rates.

Wastewater effluents cause microbial community shifts and change trophic status.

Widespread wastewater pollution is one of the greatest challenges threatening the sustainable management of rivers globally. Understanding microbial responses to gradients in environmental stressors, such as wastewater pollution, is crucial to identify thresholds of community change and to develop management strategies that protect ecosystem integrity. This study used multiple lines of empirical evidence, including a novel combination of microbial ecotoxicology methods in the laboratory and field to link pressure-stressor-response relationships. Specifically, community-based whole effluent toxicity (WET) testing and environmental genomics were integrated to determine real-world community interactions, shifts and functional change in response to wastewater pollution. Here we show that wastewater effluents above moderate (>10%) concentrations caused consistent significant shifts in bacterial community structure and function. These thresholds of community shifts were also linked to changes in the trophic state of receiving waters in terms of nutrient concentrations. Differences in the community responses along the effluent concentration gradient were primarily driven by two globally relevant bacterial indicator taxa, namely Malikia spp. (Burkholderiales) and hgcI_clade (Frankiales). Species replacement occurred above moderate effluent concentrations with abundances of Malikia spp. increasing, while abundances of hgcI_clade decreased. The responses of Malikia spp. and hgcI_clade matched gene patterns associated with globally important nitrogen cycling pathways, such as denitrification and nitrogen fixation, which linked the core individual taxa to putative function and ecosystem processes, rarely achieved in previous studies. This study has identified potential indicators of change in trophic status and the functional consequences of wastewater pollution. These findings have immediate implications for both the management of environmental stressors and protection of aquatic ecosystems.

A novel real-world ecotoxicological dataset of pelagic microbial community responses to wastewater.

Real-world observational datasets that record and quantify pressure-stressor-response linkages between effluent discharges and natural aquatic systems are rare. With global wastewater volumes increasing at unprecedented rates, it is urgent that the present dataset is available to provide the necessary information about microbial community structure and functioning. Field studies were performed at two time-points in the Austral summer. Single-species and microbial community whole effluent toxicity (WET) testing was performed at a complete range of effluent concentrations and two salinities, with accompanying environmental data to provide new insights into nutrient and organic matter cycling, and to identify ecotoxicological tipping points. The two salinity regimes were chosen to investigate future scenarios based on a predicted salinity increase at the study site, typical of coastal regions with rising sea levels globally. Flow cytometry, amplicon sequencing of 16S and 18S rRNA genes and micro-fluidic quantitative polymerase-chain reactions (MFQPCR) were used to determine chlorophyll-a and total bacterial cell numbers and size, as well as taxonomic and functional diversity of pelagic microbial communities. This strong pilot dataset could be replicated in other regions globally and would be of high value to scientists and engineers to support the next advances in microbial ecotoxicology, environmental biomonitoring and estuarine water quality modelling.