Identification of algal rich microbial blooms in the Sellafield Pile Fuel Storage Pond and the application of ultrasonic treatment to control the formation of blooms.

The presence of microorganisms in a range of nuclear facilities has been known for many years. In this study the microbial community inhabiting the Pile Fuel Storage Pond (PFSP), which is a legacy open-aired facility on the Sellafield nuclear site, Cumbria, UK, was determined to help target microbial bloom management strategies in this facility. The PFSP is currently undergoing decommissioning and the development of prolonged dense microbial blooms reduces the visibility within the water. Such impairment in the pond water visibility can lead to delays in pond operations, which also has financial implications. Efforts to control the microbial population within the PFSP are ongoing, with the installation of ultrasonic treatment units. Here next generation sequencing techniques focussing on broad targets for both eukaryotic and prokaryotic organisms were used to identify the microbial community. On-site monitoring of photosynthetic pigments indicated when microbial blooms formed and that eukaryotic algae were most likely to be responsible for these events. The sequencing data suggested that the blooms were dominated by members of the class Chrysophyceae, a group of golden algae, while evidence of cyanobacteria and other photosynthetic bacteria was limited, further supporting eukaryotic organisms causing the blooms. The results of sequencing data from 2018 was used to inform a change in the operational settings of the ultrasonic units, while monitoring of the microbial community and photosynthetic pigments trends was extended. Since the changes were made to the ultrasonic treatment, the visibility in the pond was significantly improved, with an absence of a spring bloom in 2020 and an overall reduction in the number of days lost due to microbial blooms annually. This work extends our knowledge of the diversity of microbes able to colonise nuclear fuel storage ponds, and also suggests that sequencing data can help to optimise the performance of ultrasonic treatments, to control algal proliferation in the PFSP facility and other inhospitable engineered systems.

The interplay between Cs and K in Pseudanabaena catenata; from microbial bloom control strategies to bioremediation options for radioactive waters.

Pseudanabaena dominates cyanobacterial blooms in the First-Generation Magnox Storage Pond (FGMSP) at a UK nuclear site. The fission product Cs is a radiologically significant radionuclide in the pond, and understanding the interactions between Cs and Pseudanabaena spp. is therefore important for determining facility management strategies, as well as improving understanding of microbiological responses to this non-essential chemical analogue of K. This study evaluated the fate of Cs following interactions with Pseudanabaena catenata, a laboratory strain most closely related to that dominating FGMSP blooms. Experiments showed that Cs (1 mM) exposure did not affect the growth of P. catenata, while a high concentration of K (5 mM) caused a significant reduction in cell yield. Scanning transmission X-ray microscopy elemental mapping identified Cs accumulation to discrete cytoplasmic locations within P. catenata cells, indicating a potential bioremediation option for Cs. Proteins related to stress responses and nutrient limitation (K, P) were stimulated by Cs treatment. Furthermore, selected K+ transport proteins were mis-regulated by Cs dosing, which indicates the importance of the K+ transport system for Cs accumulation. These findings enhance understanding of Cs fate and biological responses within Pseudanabaena blooms, and indicate that K exposure might provide a microbial bloom control strategy.

Isolation and identification of arbuscular mycorrhizal fungi from an abandoned uranium mine and their role in soil-to-plant transfer of radionuclides and metals.

Arbuscular mycorrhizal fungi were recovered from soil samples from the naturally radioactive soil at the long-abandoned South Terras uranium mine in Cornwall, UK. Species of Rhizophagus, Claroideoglomus, Paraglomus, Septoglomus, and Ambispora were recovered, and pot cultures from all except Ambispora were established. Cultures were identified to species level using morphological observation and rRNA gene sequencing combined with phylogenetic analysis. These cultures were used in pot experiments designed with a compartmentalised system to assess the contribution of fungal hyphae to the accumulation of essential elements, such as copper and zinc, and non-essential elements, such as lead, arsenic, thorium, and uranium into root and shoot tissues of Plantago lanceolata. The results indicated that none of the treatments had any positive or negative impact on shoot and root biomass. However, Rhizophagus irregularis treatments showed higher accumulation of copper and zinc in shoots, while R. irregularis and Septoglomus constrictum enhanced arsenic accumulation in roots. Moreover, R. irregularis increased uranium concentration in roots and shoots of the P. lanceolata plant. This study provides useful insight into fungal-plant interactions that determine metal and radionuclide transfer from soil into the biosphere at contaminated sites such as mine workings.

A three-step process of manganese acquisition and storage in the microalga Chlorella sorokiniana.

Metabolism of metals in microalgae and adaptation to metal excess are of significant environmental importance. We report a three-step mechanism that the green microalga Chlorella sorokiniana activates during the acquisition of and adaptation to manganese (Mn), which is both an essential trace metal and a pollutant of waters. In the early stage, Mn2+ was mainly bound to membrane phospholipids and phosphates in released mucilage. The outer cell wall was reorganized and lipids were accumulated, with a relative increase in lipid saturation. Intracellular redox settings were rapidly altered in the presence of Mn excess, with increased production of reactive oxygen species that resulted in lipid peroxidation and a decrease in the concentration of thiols. In the later stage, Mn2+ was chelated by polyphosphates and accumulated in the cells. The structure of the inner cell wall was modified and the redox milieu established a new balance. Polyphosphates serve as a transient Mn2+ storage ligand, as proposed previously. In the final stage, Mn was stored in multivalent Mn clusters that resemble the structure of the tetramanganese-calcium core of the oxygen-evolving complex. The present findings elucidate the bioinorganic chemistry and metabolism of Mn in microalgae, and may shed new light on water-splitting Mn clusters.

A multiscale modelling approach for Haematococcus pluvialis cultivation under different environmental conditions.

Haematococcus pluvialis can produce significant amounts of industrially important compounds belonging to lipids and starch classes, including various specific pigments such as ß-carotene, lutein and astaxanthin, as well as lipids, carbohydrates and proteins. Their production can vary depending on environmental stress conditions like nutrient starvation. However, stress conditions lead also to undesired phenomena such as cell lysis, which is likely to be related to products loss. The microorganism develops towards smaller single cell volumes during the growth process, and eventually, more likely towards lysis when fission (i.e. cell division) slows down. The lysis process takes place simultaneously with nutrient depletion, so both growth and lysis are linked to the change of environmental conditions. In this work, we develop a novel multiscale segregated-structured model based on Population Balance Equations (PBEs) to describe the photoautotrophic growth of H.pluvialis, in particular cell growth, and lysis, making possible the description of the relationship between cell volume/transition, cell loss, and metabolic product availability. Cell volume is the internal coordinate of the population balance model, and its link with intrinsic concentrations is also presented. The model parameters are fitted against experimental data, extensive sensitivity analysis is performed and the model predictive capabilities are tested in terms of cell density distributions, as well as 0th and 1st order moments.

Phylogenetic analysis and structural prediction reveal the potential functional diversity between green algae SWEET transporters.

Sugar-Will-Eventually-be-Exported-Transporters (SWEETs) are an important family of sugar transporters that appear to be ubiquitous in all organisms. Recent research has determined the structure of SWEETs in higher plants, identified specific residues required for monosaccharide or disaccharide transport, and begun to understand the specific functions of individual plant SWEET proteins. However, in green algae (Chlorophyta) these transporters are poorly characterised. This study identified SWEET proteins from across representative Chlorophyta with the aim to characterise their phylogenetic relationships and perform protein structure modelling in order to inform functional prediction. The algal genomes analysed encoded between one and six SWEET proteins, which is much less than a typical higher plant. Phylogenetic analysis identified distinct clusters of over 70 SWEET protein sequences, taken from almost 30 algal genomes. These clusters remain separate from representative higher or non-vascular plant SWEETs, but are close to fungi SWEETs. Subcellular localisation predictions and analysis of conserved amino acid residues revealed variation between SWEET proteins of different clusters, suggesting different functionality. These findings also showed conservation of key residues at the substrate-binding site, indicating a similar mechanism of substrate selectivity and transport to previously characterised higher plant monosaccharide-transporting SWEET proteins. Future work is now required to confirm the predicted sugar transport specificity and determine the functional role of these algal SWEET proteins.

Tea plant roots respond to aluminum-induced mineral nutrient imbalances by transcriptional regulation of multiple cation and anion transporters.

BACKGROUND: Tea is one of the most popular non-alcoholic beverages in the world for its flavors and numerous health benefits. The tea tree (Camellia sinensis L.) is a well-known aluminum (Al) hyperaccumulator. However, it is not fully understood how tea plants have adapted to tolerate high concentrations of Al, which causes an imbalance of mineral nutrition in the roots. RESULTS: Here, we combined ionomic and transcriptomic profiling alongside biochemical characterization, to probe the changes of metal nutrients and Al responsive genes in tea roots grown under increasing concentrations of Al. It was found that a low level of Al (~ 0.4 mM) maintains proper nutrient balance, whereas a higher Al concentration (2.5 mM) compromised tea plants by altering micro- and macro-nutrient accumulation into roots, including a decrease in calcium (Ca), manganese (Mn), and magnesium (Mg) and an increase in iron (Fe), which corresponded with oxidative stress, cellular damage, and retarded root growth. Transcriptome analysis revealed more than 1000 transporter genes that were significantly changed in expression upon Al exposure compared to control (no Al) treatments. These included transporters related to Ca and Fe uptake and translocation, while genes required for N, P, and S nutrition in roots did not significantly alter. Transporters related to organic acid secretion, together with other putative Al-tolerance genes also significantly changed in response to Al. Two of these transporters, CsALMT1 and CsALS8, were functionally tested by yeast heterologous expression and confirmed to provide Al tolerance. CONCLUSION: This study shows that tea plant roots respond to high Al-induced mineral nutrient imbalances by transcriptional regulation of both cation and anion transporters, and therefore provides new insights into Al tolerance mechanism of tea plants. The altered transporter gene expression profiles partly explain the imbalanced metal ion accumulation that occurred in the Al-stressed roots, while increases to organic acid and Al tolerance gene expression partly explains the ability of tea plants to be able to grow in high Al containing soils. The improved transcriptomic understanding of Al exposure gained here has highlighted potential gene targets for breeding or genetic engineering approaches to develop safer tea products.

Addition of organic acids to acid mine drainage polluted wetland sediment leads to microbial community structure and functional changes and improved water quality.

Acid mine drainage (AMD) is a serious environmental problem worldwide that requires efficient and sustainable remediation technologies including the use of biological mechanisms. A key challenge for AMD bioremediation is to provide optimal conditions for microbial-mediated immobilisation of trace metals. Although organic carbon and oxygen can enhance treatment efficiency, the effect on microbial communities is unclear. In this study, surface sediments from a natural wetland with proven efficiency for AMD bioremediation were artificially exposed to oxygen (by aeration) and/or organic carbon (in the form of mixed organic acids) and incubated under laboratory conditions. In addition to measuring changes in water chemistry, a metagenomics approach was used to determine changes in sediment bacterial, archaeal and fungal community structure, and functional gene abundance. The addition of organic carbon produced major changes in the abundance of microorganisms related to iron and sulfur metabolism (including Geobacter and Pelobacter) and increased levels of particulate metals via sulfate reduction. Aeration resulted in an increase in Sideroxydans abundance but no significant changes in metal chemistry were observed. The study concludes that the utilisation of organic carbon by microorganisms is more important for achieving efficient AMD treatment than the availability of oxygen, yet the combination of oxygen with organic carbon addition did not inhibit the improvements to water quality.

Optimisation of microalgal cultivation via nutrient-enhanced strategies: the biorefinery paradigm.

BACKGROUND: The production of microalgal biofuels, despite their sustainable and renowned potential, is not yet cost-effective compared to current conventional fuel technologies. However, the biorefinery concept increases the prospects of microalgal biomass as an economically viable feedstock suitable for the co-production of multiple biofuels along with value-added chemicals. To integrate biofuels production within the framework of a microalgae biorefinery, it is not only necessary to exploit multi-product platforms, but also to identify optimal microalgal cultivation strategies maximising the microalgal metabolites from which biofuels are obtained: starch and lipids. Whilst nutrient limitation is widely known for increasing starch and lipid formation, this cultivation strategy can greatly reduce microalgal growth. This work presents an optimisation framework combining predictive modelling and experimental methodologies to effectively simulate and predict microalgal growth dynamics and identify optimal cultivation strategies. RESULTS: Microalgal cultivation strategies for maximised starch and lipid formation were successfully established by developing a multi-parametric kinetic model suitable for the prediction of mixotrophic microalgal growth dynamics co-limited by nitrogen and phosphorus. The model's high predictive capacity was experimentally validated against various datasets obtained from laboratory-scale cultures of Chlamydomonas reinhardtii CCAP 11/32C subject to different initial nutrient regimes. The identified model-based optimal cultivation strategies were further validated experimentally and yielded significant increases in starch (+ 270%) and lipid (+ 74%) production against a non-optimised strategy. CONCLUSIONS: The optimised microalgal cultivation scenarios for maximised starch and lipids, as identified by the kinetic model presented here, highlight the benefits of exploiting modelling frameworks as optimisation tools that facilitate the development and commercialisation of microalgae-to-fuel technologies.

Mechanisms of detoxification of high copper concentrations by the microalga Chlorella sorokiniana.

Microalgae have evolved mechanisms to respond to changes in copper ion availability, which are very important for normal cellular function, to tolerate metal pollution of aquatic ecosystems, and for modulation of copper bioavailability and toxicity to other organisms. Knowledge and application of these mechanisms will benefit the use of microalgae in wastewater processing and biomass production, and the use of copper compounds in the suppression of harmful algal blooms. Here, using electron microscopy, synchrotron radiation-based Fourier transform infrared spectroscopy, electron paramagnetic resonance spectroscopy, and X-ray absorption fine structure spectroscopy, we show that the microalga Chlorella sorokiniana responds promptly to Cu2+ at high non-toxic concentration, by mucilage release, alterations in the architecture of the outer cell wall layer and lipid structures, and polyphosphate accumulation within mucilage matrix. The main route of copper detoxification is by Cu2+ coordination to polyphosphates in penta-coordinated geometry. The sequestrated Cu2+ was accessible and could be released by extracellular chelating agents. Finally, the reduction in Cu2+ to Cu1+ appears also to take place. These findings reveal the biochemical basis of the capacity of microalgae to adapt to high external copper concentrations and to serve as both, sinks and pools of environmental copper.

Organic complexation of U(VI) in reducing soils at a natural analogue site: Implications for uranium transport.

Understanding the long-term fate, stability, and bioavailability of uranium (U) in the environment is important for the management of nuclear legacy sites and radioactive wastes. Analysis of U behavior at natural analogue sites permits evaluation of U biogeochemistry under conditions more representative of long-term equilibrium. Here, we have used bulk geochemical and microbial community analysis of soils, coupled with X-ray absorption spectroscopy and µ-focus X-ray fluorescence mapping, to gain a mechanistic understanding of the fate of U transported into an organic-rich soil from a pitchblende vein at the UK Needle's Eye Natural Analogue site. U is highly enriched in the Needle's Eye soils (∼1600 mg kg-1). We show that this enrichment is largely controlled by U(VI) complexation with soil organic matter and not U(VI) bioreduction. Instead, organic-associated U(VI) seems to remain stable under microbially-mediated Fe(III)-reducing conditions. U(IV) (as non-crystalline U(IV)) was only observed at greater depths at the site (>25 cm); the soil here was comparatively mineral-rich, organic-poor, and sulfate-reducing/methanogenic. Furthermore, nanocrystalline UO2, an alternative product of U(VI) reduction in soils, was not observed at the site, and U did not appear to be associated with Fe-bearing minerals. Organic-rich soils appear to have the potential to impede U groundwater transport, irrespective of ambient redox conditions.

Models of microalgal cultivation for added-value products - A review.

Microalgae are considered a promising feedstock for biorefineries given that their chemical composition - rich in carbohydrate and lipid - can be directed towards the co-production of various value-added fuels and chemicals. Production of microalgal biomass for biorefinery purposes requires the identification and establishment of optimal cultivation systems, a crucial yet complicated task due to the numerous factors (e.g. media composition, light, temperature) that simultaneously regulate biomass growth and intracellular composition. Modelling these biological processes, taking into account a single or multiple growth-limiting factors, offers a valuable tool to simulate, design and optimise the dynamics of microalgae cultivation. This review provides an overview of existing models developed to describe microalgal growth processes at the macroscopic scale (also termed black-box models) and discusses their formulation in detail. The black-box kinetic modelling frameworks are compiled into single-factor (6 formulations) and multiple-factor (32 formulations - further divided into non-interactive, additive, and interactive) growth kinetic models, as reported in more than 80 studies, for the prediction of biomass growth as a function of major operational factors such as media composition (e.g. nutrient concentration) and environmental factors (e.g. transient light and temperature). In addition, the review focuses on those models that further account for the production dynamics of two microalgal intracellular products with renowned potential as biorefinery substrates: carbohydrate and lipid molecules. Models of microalgal cultivation dynamics offer a robust engineering tool to understand the natural yet complex responses of microalgae to their growing environment and can help - if used appropriately - to optimise microalgae cultivation and increase the economic viability and sustainability of microalgal systems.

The effects of ionizing radiation on the structure and antioxidative and metal-binding capacity of the cell wall of microalga Chlorella sorokiniana.

The impact of ionizing radiation on microorganisms such as microalgae is a topic of increasing importance for understanding the dynamics of aquatic ecosystems in response to environmental radiation, and for the development of efficient approaches for bioremediation of mining and nuclear power plants wastewaters. Currently, nothing is known about the effects of ionizing radiation on the microalgal cell wall, which represents the first line of defence against chemical and physical environmental stresses. Using various microscopy, spectroscopy and biochemical techniques we show that the unicellular alga Chlorella sorokiniana elicits a fast response to ionizing radiation. Within one day after irradiation with doses of 1-5 Gy, the fibrilar layer of the cell wall became thicker, the fraction of uronic acids was higher, and the capacity to remove the main reactive product of water radiolysis increased. In addition, the isolated cell wall fraction showed significant binding capacity for Cu2+, Mn2+, and Cr3+. The irradiation further increased the binding capacity for Cu2+, which appears to be mainly bound to glucosamine moieties within a chitosan-like polymer in the outer rigid layer of the wall. These results imply that the cell wall represents a dynamic structure that is involved in the protective response of microalgae to ionizing radiation. It appears that microalgae may exhibit a significant control of metal mobility in aquatic ecosystems via biosorption by the cell wall matrix.

Increased metal tolerance and bioaccumulation of zinc and cadmium in Chlamydomonas reinhardtii expressing a AtHMA4 C-terminal domain protein.

The use of microalgal biomass for metal pollutant bioremediation might be improved by genetic engineering to modify the selectivity or capacity of metal biosorption. A plant cadmium (Cd) and zinc (Zn) transporter (AtHMA4) was used as a transgene to increase the ability of Chlamydomonas reinhardtii to tolerate 0.2 mM Cd and 0.3 mM Zn exposure. The transgenic cells showed increased accumulation and internalization of both metals compared to wild-type. AtHMA4 was expressed either as the full-length (FL) protein or just the C-terminal (CT) tail, which is known to have metal-binding sites. Similar Cd and Zn tolerance and accumulation was observed with expression of either the FL protein or CT domain, suggesting that enhanced metal tolerance was mainly due to increased metal binding rather than metal transport. The effectiveness of the transgenic cells was further examined by immobilization in calcium alginate to generate microalgal beads that could be added to a metal contaminated solution. Immobilization maintained metal tolerance, while AtHMA4-expressing cells in alginate showed a concentration-dependent increase in metal biosorption that was significantly greater than alginate beads composed of wild-type cells. This demonstrates that expressing AtHMA4 FL or CT has great potential as a strategy for bioremediation using microalgal biomass.

Radiation Tolerance of Pseudanabaena catenata, a Cyanobacterium Relevant to the First Generation Magnox Storage Pond.

Recently a species of Pseudanabaena was identified as the dominant photosynthetic organism during a bloom event in a high pH (pH ∼11.4), radioactive spent nuclear fuel pond (SNFP) at the Sellafield Ltd., United Kingdom facility. The metabolic response of a laboratory culture containing the cyanobacterium Pseudanabaena catenata, a relative of the major photosynthetic microorganism found in the SNFP, to X-ray irradiation was studied to identify potential survival strategies used to support colonization of radioactive environments. Growth was monitored and the metabolic fingerprints of the cultures, during irradiation and throughout the post-irradiation recovery period, were determined using Fourier transform infrared (FT-IR) spectroscopy. A dose of 95 Gy delivered over 5 days did not significantly affect growth of P. catenata, as determined by turbidity measurements and cell counts. Multivariate statistical analysis of the FT-IR spectral data revealed metabolic variation during the post-irradiation recovery period, with increased polysaccharide and decreased amide spectral intensities. Increases in polysaccharides were confirmed by complementary analytical methods including total carbohydrate assays and calcofluor white staining. This observed increased production of polysaccharides is of significance, since this could have an impact on the fate of the radionuclide inventory in the pond via biosorption of cationic radionuclides, and may also impact on downstream processes through biofilm formation and biofouling.

Microbial bloom formation in a high pH spent nuclear fuel pond.

Microorganisms are able to colonise a wide range of extreme environments, including nuclear facilities. In this study, the First Generation Magnox Storage Pond (FGMSP) a high pH, legacy spent nuclear fuel pond (SNFP) situated at Sellafield, Cumbria, UK, was studied. Despite the inhospitable conditions in the FGMSP, microorganisms can cause "blooms" within the facility which to date have not been studied. These microbial blooms significantly reduce visibility in the engineered facility, disrupting fuel retrieval operations and slowing decommissioning. The microbial community colonising the pond during two microbial bloom periods was determined by using physiological measurements and high throughput next generation sequencing techniques. In situ probes within the ponds targeting photosynthetic pigments indicated a cyanobacterial bloom event. Analysis of the 16S rRNA gene data suggested that a single cyanobacterial genus was dominant during the bloom events, which was most closely related to Pseudanabaena sp. Comparisons between the microbial community of FGMSP and an adjacent SNFP that is periodically purged into the FGMSP, showed different community profiles. Data confirm the onset of the microbial blooms occurred when the pond purge rate was reduced, and blooms could be controlled by re-establishing the purging regime. The presence of Pseudanabaena sp. that can colonise the pond and dominate during bloom periods is notable since they have received little attention for their role in cyanobacterial bloom formation. This work also informs bioremediation efforts to treat waters contaminated with radionuclides, which could benefit from the use of cyanobacteria able to tolerate extreme environments and accumulate priority radionuclides.

Multi-genomic analysis of the cation diffusion facilitator transporters from algae.

Metal transport processes are relatively poorly understood in algae in comparison to higher plants and other eukaryotes. A screen of genomes from 33 taxonomically diverse algal species was conducted to identify members of the Cation Diffusion Facilitator (CDF) family of metal ion transporter. All algal genomes contained at least one CDF gene with four species having >10 CDF genes (median of 5 genes per genome), further confirming that this is a ubiquitous gene family. Phylogenetic analysis suggested a CDF gene organisation of five groups, which includes Zn-CDF, Fe/Zn-CDF and Mn-CDF groups, consistent with previous phylogenetic analyses, and two functionally undefined groups. One of these undefined groups was algal specific although excluded chlorophyte and rhodophyte sequences. The majority of sequences (22 out of 26 sequences) from this group had a putative ion binding site motif within transmembrane domain 2 and 5 that was distinct from other CDF proteins, such that alanine or serine replaced the conserved histidine residue. The phylogenetic grouping was supported by sequence cluster analysis. Yeast heterologous expression of CDF proteins from Chlamydomonas reinhardtii indicated Zn2+ and Co2+ transport function by CrMTP1, and Mn2+ transport function by CrMTP2, CrMTP3 and CrMTP4, which validated the phylogenetic prediction. However, the Mn-CDF protein CrMTP3 was also able to provide zinc and cobalt tolerance to the Zn- and Co-sensitive zrc1 cot1 yeast strain. There is wide diversity of CDF transporters within the algae lineage, and some of these genes may be attractive targets for future applications of metal content engineering in plants or microorganisms.

Microbial degradation of four biodegradable polymers in soil and compost demonstrating polycaprolactone as an ideal compostable plastic.

Plastics are an indispensable material but also a major environmental pollutant. In contrast, biodegradable polymers have the potential to be compostable. The biodegradation of four polymers as discs, polycaprolactone (PCL), polyhydroxybutyrate (PHB), polylactic acid (PLA) and poly(1,4 butylene) succinate (PBS) was compared in soil and compost over a period of more than 10 months at 25 °C, 37 °C and 50 °C. Degradation rates varied between the polymers and incubation temperatures but PCL showed the fastest degradation rate under all conditions and was completely degraded when buried in compost and incubated at 50 °C after 91 days. Furthermore, PCL strips showed a significant reduction in tensile strength in just 2 weeks when incubated in compost >45 °C. Various fungal strains growing on the polymer surfaces were identified by sequence analysis. Aspergillus fumigatus was most commonly found at 25 °C and 37 °C, while Thermomyces lanuginosus, which was abundant at 50 °C, was associated with PCL degradation.

Macroalgae as spatial and temporal bioindicators of coastal metal pollution following remediation and diversion of acid mine drainage.

Acid mine drainage (AMD) is a significant contributor of metal pollution leading to ecosystem damage. Bioindicator organisms such as intertidal brown macroalgae have an important role in quantifying the risks of metal bioaccumulation in coastal locations exposed to AMD contamination. Measurement of As, Cd, Cu, Fe, Pb, and Zn accumulation was performed in Fucus serratus, Fucus vesiculosus and Ascophyllum nodosum sampled from two marine locations near to an abandoned Cu mine in Anglesey, Wales, UK. Transect samples were taken from a coastal location (Amlwch) that has seen a substantial increase in AMD contamination over 15 years, in comparison to a nearby estuarine location (Dulas Estuary leading to Dulas Bay) with a historic legacy of pollution. These were compared with samples from the same sites taken 30 years earlier. Some of the Dulas macroalgae samples had Cd, Cu and Zn concentrations that were above background but in general indicated a non-polluted estuary in comparison to substantial pollution over previous decades. In contrast, Fucus samples collected from directly below an AMD outflow at Amlwch showed extremely elevated metal bioaccumulation (>250 mg Fe g-1, >6 mg Cu g-1, >2 mg Zn g-1, >190 µg As g-1) and evidence of macroalgae toxicity, indicating severe pollution at this site. However, the pollution dispersed within 200 m of the outflow source. This study has demonstrated the efficiency of three brown macroalgae species as indicators for metal bioavailability at high spatial resolution and over time.

The association of microbial activity with Fe, S and trace element distribution in sediment cores within a natural wetland polluted by acid mine drainage.

Natural recovery and remediation of acid mine drainage (AMD) reduces the generation of acidity and transport of trace elements in the runoff. A natural wetland that receives and remediates AMD from an abandoned copper mine at Parys Mountain (Anglesey, UK) was investigated for better understanding of the remediation mechanisms. Water column concentrations of dissolved Fe and S species, trace metal (loid)s and acidity decreased markedly as the mine drainage stream passed through the wetland. The metal (loid)s were removed from the water column by deposition into the sediment. Fe typically accumulated to higher concentrations in the surface layers of sediment while S and trace metal (loid)s were deposited at higher concentration within deeper (20-50 cm) sediments. High resolution X-ray fluorescence scans of sediment cores taken at three sites along the wetland indicates co-immobilization of Zn, Cu and S with sediment depth as each element showed a similar core profile. To examine the role of bacteria in sediment elemental deposition, marker genes for Fe and S metabolism were quantified. Increased expression of marker genes for S and Fe oxidation was detected at the same location within the middle of the wetland where significant decrease in SO42- and Fe2+ was observed and where generation of particulate Fe occurs. This suggests that the distribution and speciation of Fe and S that mediates the immobilization and deposition of trace elements within the natural wetland sediments is mediated in part by bacterial activity.