Cross-Phosphorylation between AgrC Histidine Kinase and the Noncognate Response Regulator Lmo1172 in Listeria monocytogenes under Benzalkonium Chloride Stress.

Benzalkonium chloride (BC) is widely used for disinfection in the food industry. However, Listeria monocytogenes strains with resistance to BC have been reported recently. In L. monocytogenes, the Agr communication system consists of a membrane-bound peptidase AgrB, a precursor peptide AgrD, a histidine kinase (HK) AgrC, and a response regulator (RR) AgrA. Our previous study showed that the agr genes are significantly upregulated by BC adaptation. This study aimed to investigate the role of the Agr system in BC resistance in L. monocytogenes. Our results showed that the Agr system was involved in BC resistance. However, a direct interaction between BC and AgrC was not observed, nor between BC and AgrA. These results indicated that BC could induce the Agr system via an indirect action. Both AgrBD and AgrC were required for growth under BC stress. Nevertheless, when exposed to BC, the gene deletion mutant ∆agrA strain exhibited better growth performance than its parental strain. The RR Lmo1172 played a role in BC resistance in the ∆agrA strain, suggesting that Lmo1172 may be an alternative to AgrA in the phosphotransfer pathway. Phosphorylation of Lmo1172 by AgrC was observed in vitro. The cognate HK Lmo1173 of Lmo1172 was not involved in BC stress, regardless of whether it was as the wild-type or the ∆agrA mutant strain. Our evidence suggests that the HK AgrC cross-phosphorylates its noncognate RR Lmo1172 to cope with BC stress when the cognate RR AgrA is absent. In vivo, further studies will be required to detect phosphotransfer of AgrC/AgrA and AgrC/Lmo1172.

Response of streamflow and nutrient loads in a small temperate catchment subject to land use change.

The aim of this study was to quantify the effect of land use change (LUC) implemented to meet nutrient load targets for a freshwater lake in New Zealand. We used the Soil and Water Assessment Tool (SWAT) model in combination with a non-parametric statistical test to determine whether afforestation of 15% of a subcatchment area was adequate to meet assigned nutrient load targets. A regional management authority set nutrient load targets of reduction in total nitrogen (TN) by 0.9 t yr-1 and reduction in total phosphorus (TP) by 0.05 t yr-1 to avoid eutrophication in the receiving waters of a freshwater lake. The load reduction was designed to be achieved through 200 ha of LUC from pasture to trees. Analysis of nutrient loads before, during, and following LUC shows that a 15% increase in forest cover decreased the annual flow (7.2%), TP load (33.3%), and TN load (13.1%). As flow and water quality observations were discrete and at irregular intervals, we used a parametric test and the SWAT model as different lines of evidence to demonstrate the effect of afforestation on flow and water quality. Policymakers concerned with decisions about LUC to improve the quality of receiving waters can benefit from applying our findings and using a statistical and numerical modelling framework to evaluate the adequacy of land use change to support improvements in water quality.

Phosphorus storage and utilization strategies of two bloom-forming freshwater cyanobacteria.

The inter-relationships between cellular phosphorus (P) storage, dissolved inorganic P (DIP) uptake affinity, alkaline phosphatase activity (APA) and dissolved inorganic nitrogen (DIN) concentrations were studied in two ubiquitous diazotrophic freshwater cyanobacteria, Raphidiopsis raciborskii (six strains) and Chrysosporum ovalisporum (two strains). DIP uptake kinetics were measured using rates of incorporation of the radio-isotope, 33P and APA as a proxy for DOP-ester utilization. The study showed that DIP uptake of individual strains followed Michaelis-Menten kinetics (modified in our study to incorporate cellular P quotas), but differed with DIN and P availability, and between growth stages. High-affinity DIP uptake and APA were activated below a P quota threshold of approximately 0.01 µg P µg-1 C across the species and strains. C. ovalisporum had significantly higher APA and P quotas (per unit C and cell) but lower uptake affinity than R. raciborskii. Demand for DIP by C. ovalisporum increased when N fixation occurred, but typically not for R. raciborskii. Our results indicate that cyanobacterial species and strains differ in their strategies to P limiting conditions, and highlight the interplay between N and P. Physiological adaptations like APA and diazotrophy of cyanobacteria adapting to low DIP and/or DIN conditions may occur simultaneously and drive species dominance in oligotrophic environments.

Plastic debris in lakes and reservoirs.

Plastic debris is thought to be widespread in freshwater ecosystems globally1. However, a lack of comprehensive and comparable data makes rigorous assessment of its distribution challenging2,3. Here we present a standardized cross-national survey that assesses the abundance and type of plastic debris (>250 µm) in freshwater ecosystems. We sample surface waters of 38 lakes and reservoirs, distributed across gradients of geographical position and limnological attributes, with the aim to identify factors associated with an increased observation of plastics. We find plastic debris in all studied lakes and reservoirs, suggesting that these ecosystems play a key role in the plastic-pollution cycle. Our results indicate that two types of lakes are particularly vulnerable to plastic contamination: lakes and reservoirs in densely populated and urbanized areas and large lakes and reservoirs with elevated deposition areas, long water-retention times and high levels of anthropogenic influence. Plastic concentrations vary widely among lakes; in the most polluted, concentrations reach or even exceed those reported in the subtropical oceanic gyres, marine areas collecting large amounts of debris4. Our findings highlight the importance of including lakes and reservoirs when addressing plastic pollution, in the context of pollution management and for the continued provision of lake ecosystem services.

A slurry approach to identify nutrient critical source areas from subtropical catchment erosion.

Targeting catchment nutrient critical source areas (CSAs) (areas contributing most of the nutrients in a catchment) is an efficient way to prioritize remediation sites for reducing nutrient runoff to waterways. We tested if the soil slurry approach - with particle sizes and sediment concentrations representative of those in streams during high rainfall events - can be used to identify potential CSAs within individual land use types, examine fire impacts, and identify the contribution of leaf litter in topsoil to nutrient export in subtropical catchments. We first confirmed the slurry approach met the prerequisite to identify CSAs with relatively higher nutrient contribution (not absolute load estimation) by comparing the slurry sampling with stream nutrient monitoring data. We validated that: 1) differences in slurry total nitrogen to phosphorus mass ratios from different land uses were consistent with stream monitoring data; and 2) our estimated nutrient export contribution from agricultural land, via the slurry approach, was comparable to that derived from monitoring data. Additionally, we found nutrient concentrations in slurries differed across soil types and management practice within individual land uses, correlating with nutrient concentrations in fine particles. These results indicate the slurry approach can be used to identify potential small-scale CSAs. Slurry results from burnt soils were also comparable to other studies showing increased levels of dissolved nutrient loss and higher nitrogen than phosphorus loss, than non-burnt soils. The slurry method also showed the contribution of leaf litter to slurry nutrient concentrations from topsoil was greater for dissolved nutrients than particulate nutrients, indicating different forms of nutrients need to be considered for impacts of vegetation. Our study reveals that the slurry method can be used to identify potential small-scale CSAs within the same land use from erosion and can account for impacts of vegetation and bushfires, providing timely information to guide catchment restoration actions.

Effects of High Ammonium Loading on Two Submersed Macrophytes of Different Growth Form Based on an 18-Month Pond Experiment.

Ammonium (NH4-N) produces a paradoxical effect on submersed macrophytes because it is not only the preferred nitrogen source for the growth of plants but also threatens the growth of plants at high concentration. Whether short-term and small-scale physiological toxicity experiments at an individual level can reflect the effects of high ammonium on populations of submersed macrophytes in natural conditions is still unclear. In this study, an 18-month experiment was conducted in six 600 m2 ponds subjected to different levels of ammonium loading. The effects of high ammonium on populations of canopy-forming Myriophyllum spicatum and rosette-forming Vallisneria natans were explored. The results showed that M. spicatum and V. natans populations can develop high cover and height at high ammonium concentration (7 mg/L) at short-term exposures, and V. natans may be tolerant to 18 mg/L ammonium concentration. However, the cover of M. spicatum and the height of both species were inhibited at 2.4 mg/L at long-term exposures. The height of M. spicatum was two to six times higher than that of V. natans across all treatments and control by the end of the experiment, and the cover of M. spicatum was 7-11 times higher than that of V. natans in most NH4-N loading treatments, except the cover of M. spicatum in the highest NH4-N loading treatment with 18 mg/L NH4-N. The rosette-forming V. natans resists ammonium stress by slow growth (shoot elongation) to reduce consumption, while canopy-forming species resist ammonium stress by shoot elongation and canopy development to capture light. Although increasing ammonium concentration may induce severe stress on M. spicatum, the morphological characteristics of this species may, to some extent, release the plants from this stress. Our present study indicates that the negative effects of ammonium stress on the development of populations increased with exposure duration, and the submersed macrophyte community with stronger ability for light capture and dispersal may resist high ammonium stress. Nevertheless, in strongly ammonium-enriched systems, competition and succession cannot be neglected.

Impacts of atmospheric stilling and climate warming on cyanobacterial blooms: An individual-based modelling approach.

Harmful algal blooms of the freshwater cyanobacteria genus Microcystis are a global problem and are expected to intensify with climate change. In studies of climate change impacts on Microcystis blooms, atmospheric stilling has not been considered. Stilling is expected to occur in some regions of the world with climate warming, and it will affect lake stratification regimes. We tested if stilling could affect water column Microcystis distributions using a novel individual-based model (IBM). Using the IBM coupled to a three-dimensional hydrodynamic model, we assessed responses of colonial Microcystis biomass to wind speed decrease and air temperature increase projected under a future climate. The IBM altered Microcystis colony size using relationships with turbulence from the literature, and included light, temperature, and nutrient effects on Microcystis growth using input data from a shallow urban lake. The model results show that dynamic variations in colony size are critical for accurate prediction of cyanobacterial bloom development and decay. Colony size (mean and variability) increased more than six-fold for a 20% decrease in wind speed compared with a 2 °C increase in air temperature. Our results suggest that atmospheric stilling needs to be included in projections of changes in the frequency, distribution and magnitude of blooms of buoyant, colony-forming cyanobacteria under climate change.

Schindler's legacy: from eutrophic lakes to the phosphorus utilization strategies of cyanobacteria.

David Schindler and his colleagues pioneered studies in the 1970s on the role of phosphorus in stimulating cyanobacterial blooms in North American lakes. Our understanding of the nuances of phosphorus utilization by cyanobacteria has evolved since that time. We review the phosphorus utilization strategies used by cyanobacteria, such as use of organic forms, alternation between passive and active uptake, and luxury storage. While many aspects of physiological responses to phosphorus of cyanobacteria have been measured, our understanding of the critical processes that drive species diversity, adaptation and competition remains limited. We identify persistent critical knowledge gaps, particularly on the adaptation of cyanobacteria to low nutrient concentrations. We propose that traditional discipline-specific studies be adapted and expanded to encompass innovative new methodologies and take advantage of interdisciplinary opportunities among physiologists, molecular biologists, and modellers, to advance our understanding and prediction of toxic cyanobacteria, and ultimately to mitigate the occurrence of blooms.

Chlorophyll and phycocyanin in-situ fluorescence in mixed cyanobacterial species assemblages: Effects of morphology, cell size and growth phase.

Cyanobacteria harmful blooms can represent a major risk for public health due to potential release of toxins and other noxious compounds in the water. A continuous and high-resolution monitoring of the cyanobacteria population is required due to their rapid dynamics, which has been increasingly done using in-situ fluorescence of phycocyanin (f-PC) and chlorophyll a (f-Chl a). Appropriate in-situ fluorometers calibration is essential because f-PC and f-Chl a are affected by biotic and abiotic factors, including species composition. Measurement of f-PC and f-Chl a in mixed species assemblages during different growth phases - representative of most field conditions - has received little attention. We hypothesized that f-PC and f-Chl a of mixed assemblages of cyanobacteria may be accurately estimated if taxa composition and fluorescence characteristics are known. We also hypothesized that species with different morphologies would have different fluorescence per unit cell and biomass. We tested these hypotheses in a controlled culture experiment in which photosynthetic pigment fluorescence, chemical pigment extraction, optical density and microscopic enumeration of four common cyanobacteria species (Aphanocapsa sp, Microcystis aeruginosa, Dolichospermum circinale and Raphidiopsis raciborskii) were quantified. Both monocultures and mixed cultures were monitored from exponential to late stationary growth phases. The sum of fluorescence of individual species calculated for mixed samples was not significantly different than measured fluorescence of mixed cultures. Estimated and measured f-PC and f-Chl a of mixed cultures had higher correlations and smaller absolute median errors when estimations were based on fluorescence per biomass instead of fluorescence per cell. Largest errors were overestimations of measured fluorescence for species with different morphologies. Fluorescence per cell was significantly different among most species, while fluorescence per unit biomass was not, indicating that conversion of fluorescence to biomass reduces species-specific bias. This study presents new information on the effect of species composition on cyanobacteria fluorescence. Best practices of deployment and operation of fluorometers, and data-driven models supporting in-situ fluorometers calibration are discussed as suitable solutions to minimize taxa-specific bias in fluorescence estimates.

Polluted lake restoration to promote sustainability in the Yangtze River Basin, China.

China has made a concerted effort to successfully improve water quality of rivers, but lake water quality has not improved. Lakes require controls on both catchment external nutrient loads and in-lake internal loads, where nature-based solutions are coupled with engineered systems to achieve the United Nations Sustainable Development Goals (SDGs).

A bacterial index to estimate lake trophic level: National scale validation.

Lakes and their catchments have been subjected to centuries to millennia of exploitation by humans. Efficient monitoring methods are required to promote proactive protection and management. Traditional monitoring is time consuming and expensive, which limits the number of lakes monitored. Lake surface sediments provide a temporally integrated representation of environmental conditions and contain high microbial biomass. Based on these attributes, we hypothesized that bacteria associated with lake trophic states could be identified and used to develop an index that would not be confounded by non-nutrient stressor gradients. Metabarcoding (16S rRNA gene) was used to assess bacterial communities present in surface sediments from 259 non-saline lakes in New Zealand encompassing a range of trophic states from alpine microtrophic lakes to lowland hypertrophic lakes. A subset of lakes (n = 96) with monitoring data was used to identify indicator amplicon sequence variants (ASVs) associated with different trophic states. A total of 10,888 indicator taxa were identified and used to develop a Sediment Bacterial Trophic Index (SBTI), which signficantly correlated (r2 = 0.842, P < 0.001) with the Trophic Lake Index. The SBTI was then derived for the remaining 163 lakes, providing new knowledge of the trophic state of these unmonitored lakes. This new, robust DNA-based tool provides a rapid and cost-effective method that will allow a greater number of lakes to be monitored and more effectively managed in New Zealand and globally. The SBTI could also be applied in a paleolimnological context to investigate changes in trophic status over centuries to millennia.

Submersed macrophyte restoration with artificial light-emitting diodes: A mesocosm experiment.

Urban lakes are important natural assets but are exposed to multiple stressors from human activities. Submersed macrophytes, a key plant group that helps to maintain clear-water conditions in lakes, tend to be scarce in urban lakes, particularly when they are eutrophic or hypertrophic, and their loss is linked, in part, to impaired underwater light climate. We tested if enhancing the underwater light conditions using light-emitting diodes (LEDs) could restore submersed macrophytes in urban lakes. Twelve mesocosms (1000 L each) were each planted with tape grass (Vallisneria natans) and monitored over three months (22 August-7 November), using a control and three artificial light intensity treatments (10, 50, and 100 µmol m-2 s-1). Compared with the control, the high light treatment (100 µmol m-2 s-1) had higher leaf number, maximum leaf length, and average leaf length (3.9, 5.8, and 2.8 times, respectively). Shoot number, leaf number, leaf dry mass, root dry mass, and photosynthetic photon flux density in the high-light treatment were significantly greater than the control, but root length and phytoplankton chlorophyll a were not related to plant growth variables and were low in all treatments. Periphyton chlorophyll a increased significantly with the plant growth variables (i.e., shoot number, leaf number, and maximum leaf length) and was high in the light treatments but did not hamper the growth of the macrophytes. These results indicate that LED light supplementation enables the growth of V. natans under eutrophic conditions, at least in the absence of fish as in our experiment, and that the method may have potential as a restoration method in urban lakes. Lake-scale studies are needed, however, to fully evaluate LED light supplementation under natural conditions where other stressors (e.g., fish grazing) may need to be controlled for successful restoration of urban lakes.

Global data set of long-term summertime vertical temperature profiles in 153 lakes.

Climate change and other anthropogenic stressors have led to long-term changes in the thermal structure, including surface temperatures, deepwater temperatures, and vertical thermal gradients, in many lakes around the world. Though many studies highlight warming of surface water temperatures in lakes worldwide, less is known about long-term trends in full vertical thermal structure and deepwater temperatures, which have been changing less consistently in both direction and magnitude. Here, we present a globally-expansive data set of summertime in-situ vertical temperature profiles from 153 lakes, with one time series beginning as early as 1894. We also compiled lake geographic, morphometric, and water quality variables that can influence vertical thermal structure through a variety of potential mechanisms in these lakes. These long-term time series of vertical temperature profiles and corresponding lake characteristics serve as valuable data to help understand changes and drivers of lake thermal structure in a time of rapid global and ecological change.

Variability in microcystin quotas during a Microcystis bloom in a eutrophic lake.

Microcystis is a bloom-forming genus of cyanobacteria with some genotypes that produce highly toxic microcystin hepatotoxins. In waterbodies where biological and physical factors are relatively homogenous, toxin quotas (the average amount of toxin per cell), at a single point in time, are expected to be relatively constant. In this study we challenged this assumption by investigating the spatial distribution of microcystin quotas at a single point in time on two separate occasions in a lake with a major Microcystis bloom. Microcystis cell concentrations varied widely across the lake on both sampling occasions (730- and 137-fold) together with microcystin quotas (148- and 362-fold). Cell concentrations and microcystin quotas were strongly positively correlated (R2 = 0.89, P < 0.001, n = 28; R2 = 0.67, P < 0.001, n = 25). Analysis of Microcystis strains using high-throughput sequencing of the 16S-23S rRNA intergenic spacer region showed no relationship between microcystin quota and the relative abundance of specific sequences. Collectively, the results of this study indicate an association between microcystin production and cell density that magnifies the potential for bloom toxicity at elevated cell concentrations.

Global co-occurrence of methanogenic archaea and methanotrophic bacteria in Microcystis aggregates.

Global warming and eutrophication contribute to the worldwide increase in cyanobacterial blooms, and the level of cyanobacterial biomass is strongly associated with rises in methane emissions from surface lake waters. Hence, methane-metabolizing microorganisms may be important for modulating carbon flow in cyanobacterial blooms. Here, we surveyed methanogenic and methanotrophic communities associated with floating Microcystis aggregates in 10 lakes spanning four continents, through sequencing of 16S rRNA and functional marker genes. Methanogenic archaea (mainly Methanoregula and Methanosaeta) were detectable in 5 of the 10 lakes and constituted the majority (~50%-90%) of the archaeal community in these lakes. Three of the 10 lakes contained relatively more abundant methanotrophs than the other seven lakes, with the methanotrophic genera Methyloparacoccus, Crenothrix, and an uncultured species related to Methylobacter dominating and nearly exclusively found in each of those three lakes. These three are among the five lakes in which methanogens were observed. Operational taxonomic unit (OTU) richness and abundance of methanotrophs were strongly positively correlated with those of methanogens, suggesting that their activities may be coupled. These Microcystis-aggregate-associated methanotrophs may be responsible for a hitherto overlooked sink for methane in surface freshwaters, and their co-occurrence with methanogens sheds light on the methane cycle in cyanobacterial aggregates.

Individual-based modelling of cyanobacteria blooms: Physical and physiological processes.

Lakes and reservoirs throughout the world are increasingly adversely affected by cyanobacterial harmful algal blooms (CyanoHABs). The development and spatiotemporal distributions of blooms are governed by complex physical mixing and transport processes that interact with physiological processes affecting the growth and loss of bloom-forming species. Individual-based models (IBMs) can provide a valuable tool for exploring and integrating some of these processes. Here we contend that the advantages of IBMs have not been fully exploited. The main reasons for the lack of progress in mainstreaming IBMs in numerical modelling are their complexity and high computational demand. In this review, we identify gaps and challenges in the use of IBMs for modelling CyanoHABs and provide an overview of the processes that should be considered for simulating the spatial and temporal distributions of cyanobacteria. Notably, important processes affecting cyanobacteria distributions, in particular their vertical passive movement, have not been considered in many existing lake ecosystem models. We identify the following research gaps that should be addressed in future studies that use IBMs: 1) effects of vertical movement and physiological processes relevant to cyanobacteria growth and accumulations, 2) effects and feedbacks of CyanoHABs on their environment; 3) inter and intra-specific competition of cyanobacteria species for nutrients and light; 4) use of high resolved temporal-spatial data for calibration and verification targets for IBMs; and 5) climate change impacts on the frequency, intensity and duration of CyanoHABs. IBMs are well adapted to incorporate these processes and should be considered as the next generation of models for simulating CyanoHABs.

Light-induced fluorescence quenching leads to errors in sensor measurements of phytoplankton chlorophyll and phycocyanin.

Optical sensors for fluorescence of chlorophyll a (f-Chl a) and phycocyanin (f-PC) are increasingly used as a proxy for biomass of algae and cyanobacteria, respectively. They provide measurements at high-frequency and modest cost. These sensors require site-specific calibration due to a range of interferences. Light intensity affects the fluorescence yield of cyanobacteria and algae through light harvesting regulation mechanisms, but is often neglected as a potential source of error for in-situ f-Chl a and f-PC measurements. We hypothesised that diel light variations would induce significant f-Chl a and f-PC suppression when compared to dark periods. We tested this hypothesis in a controlled experiment using three commercial fluorescence probes which continuously measured f-Chl a and f-PC from a culture of the cyanobacterium Dolichospermum variabilis as well as f-Chl a from a culture of the green alga Ankistrodesmus gracilis in a simulated natural light regime. Under light, all devices showed a significant (p<0.01) suppression of f-Chl a and f-PC compared to measurements in the dark. f-Chl a decreased by up to 79% and f-PC by up to 59% at maximum irradiance compared to dark-adapted periods. Suppression levels were higher during the second phase of the diel cycle (declining light), indicating that quenching is dependent on previous light exposure. Diel variations in light intensity must be considered as a significant source of bias for fluorescence probes used for algal monitoring. This is of high relevance as most monitoring activities take place during daytime and hence f-Chl a and f-PC are likely to be systematically underestimated under bright conditions. Compensation models, design modifications to fluorometers and sampling design are discussed as suitable alternatives to overcome light-induced fluorescence quenching.

Deeper waters are changing less consistently than surface waters in a global analysis of 102 lakes.

Globally, lake surface water temperatures have warmed rapidly relative to air temperatures, but changes in deepwater temperatures and vertical thermal structure are still largely unknown. We have compiled the most comprehensive data set to date of long-term (1970-2009) summertime vertical temperature profiles in lakes across the world to examine trends and drivers of whole-lake vertical thermal structure. We found significant increases in surface water temperatures across lakes at an average rate of + 0.37 °C decade-1, comparable to changes reported previously for other lakes, and similarly consistent trends of increasing water column stability (+ 0.08 kg m-3 decade-1). In contrast, however, deepwater temperature trends showed little change on average (+ 0.06 °C decade-1), but had high variability across lakes, with trends in individual lakes ranging from - 0.68 °C decade-1 to + 0.65 °C decade-1. The variability in deepwater temperature trends was not explained by trends in either surface water temperatures or thermal stability within lakes, and only 8.4% was explained by lake thermal region or local lake characteristics in a random forest analysis. These findings suggest that external drivers beyond our tested lake characteristics are important in explaining long-term trends in thermal structure, such as local to regional climate patterns or additional external anthropogenic influences.

A systematic literature review of forecasting and predictive models for cyanobacteria blooms in freshwater lakes.

Cyanobacteria harmful blooms (CyanoHABs) in lakes and reservoirs represent a major risk for water authorities globally due to their toxicity and economic impacts. Anticipating bloom occurrence and understanding the main drivers of CyanoHABs are needed to optimize water resources management. An extensive review of the application of CyanoHABs forecasting and predictive models was performed, and a summary of the current state of knowledge, limitations and research opportunities on this topic is provided through analysis of case studies. Two modelling approaches were used to achieve CyanoHABs anticipation; process-based (PB) and data-driven (DD) models. The objective of the model was a determining factor for the choice of modelling approach. PB models were more frequently used to predict future scenarios whereas DD models were employed for short-term forecasts. Each modelling approach presented multiple variations that may be applied for more specific, targeted purposes. Most models reviewed were site-specific. The monitoring methodologies, including data frequency, uncertainty and precision, were identified as a major limitation to improve model performance. A lack of standardization of both model output and performance metrics was observed. CyanoHAB modelling is an interdisciplinary topic and communication between disciplines should be improved to facilitate model comparisons. These shortcomings can hinder the adoption of modelling tools by practitioners. We suggest that water managers should focus on generalising models for lakes with similar characteristics and where possible use high frequency monitoring for model development and validation.