Improper Maintenance Activities Alter Benefits of Urban Stormwater Treatment in a Temperate Constructed Wetland in NSW, Australia

Constructed wetlands (CWs) are an effective means to treat nutrient and sediment pollution in urban stormwater runoff to minimise impact on receiving waterways. Maintenance of devices is recognised as a major contributing factor to performance. There is a lack of evidence-based guidance on maintenance activities to optimise treatment, due to a paucity of data from long-term field studies into CW performance before and after maintenance. In this study, the nutrient and sediment removal efficiency (% RE) of a CW was evaluated by calculating removal efficiencies of nitrogen (N), phosphorus (P) and total suspended sediment (TSS) following a long-term sampling program under baseflow and event flow conditions. Sampling was carried out before, during and after maintenance. Maintenance involved removing all aquatic vegetation and 200–300 mm of sediments over a 3-week period, aiming to improve the wetland’s performance. Assessment of dissolved and particulate nutrient fractions allowed a comprehensive investigation into drivers of nutrient removal efficiency. Under baseflow conditions differences in inflow and outflow pollutant concentrations were used to calculate removal efficiency and pollutant loads were used during event flow conditions. Before maintenance, during baseflow conditions the wetland was removing total N (36% RE) but exporting total P (-52% RE) and total sediment (-94% RE). During event-flow conditions all target pollutants were being removed (total N 63% RE, total P 25% RE and TSS 69% RE). phosphorusDuring maintenance, the device continued to remove total N (18% RE) but the physical disturbance of the maintenance resulted in mass export of total P (-120% RE) and total sediment (−2,000% RE) over a short time period, effectively undoing previous treatment. After maintenance, during baseflow conditions, the wetlands’ ability to treat total N decreased (28% RE), improved for total P (1% RE), and became a chronic source of suspended sediment (−127% RE). During event flow conditions, total N was no longer being treated (−19%) but total P and total suspended sediment were being retained (74%, 80% RE respectively). This study showed that the physical disturbance resulting from large-scale maintenance activities can potentially reverse years’ worth of treatment if not adequately planned and carried out with suitable controls

Key drivers of seasonal plankton dynamics in cyclonic and anticyclonic eddies off East Australia

© 2016 Laiolo, McInnes, Matear and Doblin. Mesoscale eddies in the south west Pacific region are prominent ocean features that represent distinctive environments for phytoplankton. Here, we examine the seasonal plankton dynamics associated with averaged cyclonic and anticyclonic eddies (CE and ACE, respectively) off eastern Australia. We do this through building seasonal climatologies of mixed layer depth (MLD) and surface chlorophyll-a for both CE and ACE by combining remotely sensed sea surface height (TOPEX/Poseidon, Envisat, Jason-1, and OSTM/Jason-2), remotely sensed ocean color (GlobColour) and in situ profiles of temperature, salinity and pressure from Argo floats. Using the CE and ACE seasonal climatologies, we assimilate the surface chlorophyll-a data into both a single (WOMBAT), and multi-phytoplankton class (EMS) biogeochemical model to investigate the level of complexity required to simulate the phytoplankton chlorophyll-a. For the two eddy types, the data assimilation showed both biogeochemical models only needed one set of parameters to represent phytoplankton but needed different parameters for zooplankton. To assess the simulated phytoplankton behavior we compared EMS model simulations with a ship-based experiment that involved incubating a winter phytoplankton community sampled from below the mixed layer under ambient and two higher light intensities with and without nutrient enrichment. By the end of the 5-day field experiment, large diatom abundance was four times greater in all treatments compared to the initial community, with a corresponding decline in pico-cyanobacteria. The experimental results were consistent with the simulated behavior in CE and ACE, where the seasonal deepening of the mixed layer during winter produced a rapid increase in large phytoplankton. Our model simulations suggest that CE off East Australia are not only characterized by a higher chlorophyll-a concentration compared to ACE, but also by a higher concentration of large phytoplankton (i.e., diatoms) due to the shallower CE mixed layer. The model simulations also suggest the zooplankton community is different in the two eddy types and this behavior needs further investigation

Predictability of thermal fluctuations influences functional traits of a cosmopolitan marine diatom

Evolutionary theory predicts that organismal plasticity should evolve in environments that fluctuate regularly. However, in environments that fluctuate less predictably, plasticity may be constrained because environmental cues become less reliable for expressing the optimum phenotype. Here, we examine how the predictability of +5°C temperature fluctuations impacts the phenotype of the marine diatom Thalassiosira pseudonana. Thermal regimes were informed by temperatures experienced by microbes in an ocean simulation and featured regular or irregular temporal sequences of fluctuations that induced mild physiological stress. Physiological traits (growth, cell size, complexity and pigmentation) were quantified at the individual cell level using flow cytometry. Changes in cellular complexity emerged as the first impact of predictability after only 8–11 days, followed by deleterious impacts on growth on days 13–16. Specifically, cells with a history of irregular fluctuation exposure exhibited a 50% reduction in growth compared with the stable reference environment, while growth was 3–18 times higher when fluctuations were regular. We observed no evidence of heat hardening (increasingly positive growth) with recurrent fluctuations. This study demonstrates that unpredictable temperature fluctuations impact this cosmopolitan diatom under ecologically relevant time frames, suggesting shifts in environmental stochasticity under a changing climate could have widespread consequences among ocean primary producers

A high-throughput assay for quantifying phenotypic traits of microalgae

High-throughput methods for phenotyping microalgae are in demand across a variety of research and commercial purposes. Many microalgae can be readily cultivated in multi-well plates for experimental studies which can reduce overall costs, while measuring traits from low volume samples can reduce handling. Here we develop a high-throughput quantitative phenotypic assay (QPA) that can be used to phenotype microalgae grown in multi-well plates. The QPA integrates 10 low-volume, relatively high-throughput trait measurements (growth rate, cell size, granularity, chlorophyll a, neutral lipid content, silicification, reactive oxygen species accumulation, and photophysiology parameters: ETRmax, Ik, and alpha) into one workflow. We demonstrate the utility of the QPA on Thalassiosira spp., a cosmopolitan marine diatom, phenotyping six strains in a standard nutrient rich environment (f/2 media) using the full 10-trait assay. The multivariate phenotypes of strains can be simplified into two dimensions using principal component analysis, generating a trait-scape. We determine that traits show a consistent pattern when grown in small volume compared to more typical large volumes. The QPA can thus be used for quantifying traits across different growth environments without requiring exhaustive large-scale culturing experiments, which facilitates experiments on trait plasticity. We confirm that this assay can be used to phenotype newly isolated diatom strains within 4 weeks of isolation. The QPA described here is highly amenable to customisation for other traits or unicellular taxa and provides a framework for designing high-throughput experiments. This method will have applications in experimental evolution, modelling, and for commercial applications where screening of phytoplankton traits is of high importance

Multitrait diversification in marine diatoms in constant and warmed environments

Phytoplankton are photosynthetic marine microbes that affect food webs, nutrient cycles and climate regulation. Their roles are determined by correlated phytoplankton functional traits including cell size, chlorophyll content and cellular composition. Here, we explore patterns of evolution in interrelated trait values and correlations. Because both chance events and natural selection contribute to phytoplankton trait evolution, we used population bottlenecks to diversify six genotypes of Thalassiosirid diatoms. We then evolved them as large populations in two environments. Interspecific variation and within-species evolution were visualized for nine traits and their correlations using reduced axes (a trait-scape). Our main findings are that shifts in trait values resulted in movement of evolving populations within the trait-scape in both environments, but were more frequent when large populations evolved in a novel environment. Which trait relationships evolved was population-specific, but greater departures from ancestral trait correlations were associated with lower population growth rates. There was no single master trait that could be used to understand multi-trait evolution. Instead, repeatable multi-trait evolution occurred along a major axis of variation defined by several diatom traits and trait relationships. Because trait-scapes capture changes in trait relation-ships and values together, they offer an insightful way to study multi-trait variatio

The evolution of trait correlations constrains phenotypic adaptation to high CO 2 in a eukaryotic alga

Microbes form the base of food webs and drive biogeochemical cycling. Predicting the effects of microbial evolution on global elemental cycles remains a significant challenge due to the sheer number of interacting environmental and trait combinations. Here, we present an approach for integrating multivariate trait data into a predictive model of trait evolution. We investigated the outcome of thousands of possible adaptive walks parameterized using empirical evolution data from the alga Chlamydomonas exposed to high CO(2). We found that the direction of historical bias (existing trait correlations) influenced both the rate of adaptation and the evolved phenotypes (trait combinations). Critically, we use fitness landscapes derived directly from empirical trait values to capture known evolutionary phenomena. This work demonstrates that ecological models need to represent both changes in traits and changes in the correlation between traits in order to accurately capture phytoplankton evolution and predict future shifts in elemental cycling

Mucospheres produced by a mixotrophic protist impact ocean carbon cycling

Mixotrophic protists (unicellular eukaryotes) that engage in both phototrophy (photosynthesis) and phago-heterotrophy (engulfment of particles)-are predicted to contribute substantially to energy fluxes and marine biogeochemical cycles. However, their impact remains largely unquantified. Here we describe the sophisticated foraging strategy of a widespread mixotrophic dinoflagellate, involving the production of carbon-rich 'mucospheres' that attract, capture, and immobilise microbial prey facilitating their consumption. We provide a detailed characterisation of this previously undescribed behaviour and reveal that it represents an overlooked, yet quantitatively significant mechanism for oceanic carbon fluxes. Following feeding, the mucospheres laden with surplus prey are discarded and sink, contributing an estimated 0.17-1.24 mg m-2 d-1 of particulate organic carbon, or 0.02-0.15 Gt to the biological pump annually, which represents 0.1-0.7% of the estimated total export from the euphotic zone. These findings demonstrate how the complex foraging behaviour of a single species of mixotrophic protist can disproportionally contribute to the vertical flux of carbon in the ocean

Colorimetric Detection of Caspase 3 Activity and Reactive Oxygen Derivatives: Potential Early Indicators of Thermal Stress in Corals

There is an urgent need to develop and implement rapid assessments of coral health to allow effective adaptive management in response to coastal development and global change. There is now increasing evidence that activation of caspase-dependent apoptosis plays a key role during coral bleaching and subsequent mortality. In this study, a "clinical" approach was used to assess coral health by measuring the activity of caspase 3 using a commercial kit. This method was first applied while inducing thermal bleaching in two coral species, Acropora millepora and Pocillopora damicornis. The latter species was then chosen to undergo further studies combining the detection of oxidative stress-related compounds (catalase activity and glutathione concentrations) as well as caspase activity during both stress and recovery phases. Zooxanthellae photosystem II (PSII) efficiency and cell density were measured in parallel to assess symbiont health. Our results demonstrate that the increased caspase 3 activity in the coral host could be detected before observing any significant decrease in the photochemical efficiency of PSII in the algal symbionts and/or their expulsion from the host. This study highlights the potential of host caspase 3 and reactive oxygen species scavenging activities as early indicators of stress in individual coral colonies

New resource for population genetics studies on the Australasian intertidal brown alga, Hormosira banksii: isolation and characterization of 15 polymorphic microsatellite loci through next generation DNA sequencing

The Australasian fucoid, Hormosira banksii, commonly known as ‘Neptune’s necklace’ or ‘bubbleweed’ is regarded as an autogenic ecosystem engineer with no functional equivalents. Population declines resulting from climate change and other anthropogenic disturbances pose significant threats to intertidal biodiversity. For effective conservation strategies, patterns of gene flow and population genetic structure across the species distribution need to be clearly understood. We developed a suite of 15 polymorphic microsatellite markers using next generation sequencing of 53–55 individuals from two sites (south-western Victoria and central New South Wales, Australia) and a replicated spatially hierarchical sampling design. We observed low to moderate genetic variation across most loci (mean number of alleles per locus =3.26; mean expected heterozygosity =0.38) with no evidence of individual loci deviating significantly from Hardy-Weinberg equilibrium. Marker independence was confirmed with tests for linkage disequilibrium, and analyses indicated no evidence of null alleles across loci. Independent spatial autocorrelation analyses were performed for each site using multilocus genotypes and different relatedness measures. Both analyses indicated no significant patterns between relatedness and geographic distance, complemented by non-significant Hardy-Weinberg estimates (P < 0.05), suggesting that individuals from each site represent a randomly mating, outcrossing population. A preliminary investigation of population structure indicates that gene flow among sites is limited (FST = 0.49), however more comprehensive sampling is needed to determine the extent of population structure across the species range (>10,000 km). The genetic markers described provide a valuable resource for future population genetic assessments that will help guide conservation planning for H. banksii and the associated intertidal communities