The Arctic is a region undergoing unprecedented and unequivocal climate change. The seas of this extreme region form a major component of the oceanic thermohaline conveyor and natural carbon cycle. Using a combination of recent and historical datasets this study examines the distribution, diversity, photophysiology and primary productivity of phytoplankton in the Atlantic sector of the Arctic Ocean. CHEMTAX analysis reveals a diverse phytoplankton community structure in the Greenland Sea comprising six main phytoplankton groups. The influence of sea-ice and water column stratification are key factors in the presence or absence of groups such as haptophytes and prasinophytes. Group-specific differences are observed in spectral absorption and photophysiological parameters. However, the influence of environmental factors has a stronger influence than taxonomic composition on photophysiology. A clear division between the photoacclimatory response of algal communities beneath sea-ice and those of open-ocean stations is predominantly due to ‘E_k independent’ photoacclimation beneath sea-ice. This occurs due to the combined effect of sea-ice decreasing irradiance entering the water column and a positive correlation between P_m ^B and temperature. This variation in photophysiology is important for primary production models as a sensitivity analysis shows that errors in these parameters propagate to give the largest final errors in primary production values. The importance of other model parameters varies with the level of biomass in the water column and the presence or absence of sea ice. Accelerated ice-melt and an increase in open water due to climate change are likely to increase primary production in the Atlantic Arctic alongside an altered distribution of phytoplankton groups, with an increase in the importance of prasinophytes or haptophytes.

Dimethylsulphoniopropionate (DMSP) is a dimethylated sulphur compound that appears to be produced by most marine algae and is a major component of the marine sulphur cycle. The majority of research to date has focused on the production of DMSP and its major breakdown product, the climatically important gas dimethylsulphide (DMS) (collectively DMS/P), by phytoplankton in the open ocean. A number of functions for intracellular DMSP (DMSPi) in phytoplankton have been identified and the cycling of DMS/P appears to be critical for ecosystem function. However, mechanisms for the production and release of DMS/P in the coastal ocean are poorly understood, despite the region’s economic and ecological importance. Coralline algal habitats (e.g. maerl beds, coral reefs, seagrass meadows, kelp forests) are distributed throughout the coastal oceans worldwide. Their three-dimensional structure supports high biodiversity and provides numerous services, generating considerable economic wealth. DMSPi in coralline algae is known to be high, thus coralline algal habitats may be critical components of the coastal sulphur cycle. This research aimed to improve our understanding of the production of DMS/P by coralline algal habitats by investigating (1) natural spatiotemporal variation and (2) the influence of environmental pressures. This was achieved through a number of laboratory and field-based studies, utilising modern and well-established techniques. The first objective of this research was to better understand the photosynthesis of red coralline algae (Chapter 3), as the algal precursor to DMSPi is methionine, a product of photosynthesis. The photosynthetic characteristics of coralline algae exhibited acclimation to changing light conditions (e.g. over a diurnal cycle or between natural and static lighting conditions). Further, for the species tested, coralline algae are often subjected to light-saturating natural conditions, therefore requiring efficient photo-protective mechanisms, which may include DMSPi regulation. On a global scale, DMSPi in coralline algae may decline with latitude, reinforcing the role of DMSPi as an antioxidant (Chapter 4). At smaller spatial scales, DMS/P production, release and recycling mechanisms were apparent in a number of habitat types (Chapter 4). A strong seasonal trend in DMS/P was also observed at a Scottish maerl bed, driven by water temperature and cloud cover (Chapter 5). Annually averaged DMS and DMSP concentrations were 230% and 700% respectively higher than the open ocean, highlighting the potential importance of the coastal ocean in the marine sulphur cycle (Chapter 5). The influence of environmental pressures (decreased salinity, variable pH and grazing) on DMS/P production by coralline algal habitats was examined (Chapters 6 – 8). In agreement with the phytoplankton literature, a chronic, but not acute, reduction in salinity led to a significant decline in coralline algal DMSPi concentrations and a sinking of the surface epithelial cells but no apparent impact on photosynthesis (Chapter 6). In the naturally variable tropical reef environment, calcifying algae continually regulated DMSPi concentrations in response to the diurnal cycling of carbonate saturation state (Chapter 7), suggesting that DMSPi may be enhanced under low pH regimes to compensate for enhanced oxidant production. Under low pH conditions, cracks were observed between the surface epithelial cells of coralline algae, potentially allowing DMSPi to leak from the cells (Chapter 7). In the field, grazing by urchins appeared to facilitate the release of DMS/P from kelp in coralline algal habitats (Chapter 8). In the laboratory, DMSPi in coralline algae increased in response to chemical cues from grazers rather than direct grazing activity, as had been previously proposed. Prior to this research, little information was available on DMS/P concentrations in coralline algal habitats. The marine sulphur cycle may impact climate regulation and ecosystem function on a global scale. This research provides a comprehensive source of information on the importance of coralline algal habitats in the marine sulphur cycle by examining natural variability and potential changes in response to environmental perturbations. This work will form a baseline for continued research in this field, investigating, for example, the impact of multiple stressors on DMS/P production, release and recycling in coastal marine habitats.

This work investigates, for the first time, the Pseudo-nitzschia (PN) dynamics in the western English Channel (L4) and the environmental factors impacting on domoic acid (DA) production in these waters. This is combined with laboratory studies examining key environmental factors and the multifactorial impact of multiple macronutrient and micronutrient availability on PN growth and DA production. An LC-MS method was established, optimised, and compared with ELISA for the accurate and reproducible extraction and determination of particulate and dissolved DA. The method was used to measure the seasonal variation in DA at L4 during 2009 and this was compared to PN seasonal abundance and diversity. Three groups a P. delicatissima-group, a P. seriata-group, and a P. pungens/multiseries-group were identified and were found to have different ecological distributions with the latter two groups significantly correlating with DA concentration. Macronutrients, in combination with other environmental factors, were found to influence PN populations at L4. Multifactorial laboratory culture experiments investigating the availability of nitrate, phosphate, and silicate, confirmed that the interrelatedness of all these nutrients significantly affected the growth, decline, and DA production of P. multiseries, and highlight the importance of both phosphate and silicate availability for DA production. When the impacts of both macronutrient (phosphate and silicate) and micronutrient (iron and copper) availability were investigated, limited growth and DA production was observed in P. multiseries cultures. Results revealed the complexity and interrelationship of factors affecting both PN growth and DA production. Furthermore, molecular methods were developed to elucidate the PN species present from 2009 Lugol’s-preserved L4 samples. DNA was successfully extracted and amplified from these samples which had been stored for up to 2 years. Initial sequence analysis identified the rbcL DNA marker as an informative site for future work with a number of L4 sequences closely relating to different Pseudo-nitzschia spp.

Phytoplankton play a key role in the geochemical cycles of the Earth, are responsible for 50% of global carbon fixation, and through this, provide almost all of the energy for the entire marine trophic system. Understanding the dynamics of phytoplankton, and the species composition in relation to environmental factors is therefore of great importance. In this thesis a range of techniques to identify phytoplankton groups that use accessory pigment data obtained from high performance liquid chromatography are compared. While fixed indicator pigment:chl-a ratio approaches provide a quick and simple way of estimating phytoplankton distributions either on the basis of size-class or taxonomic group, the more sophisticated iterative approach of CHEMTAX divides the biomass into more categories and allows more flexibility to adapt to changes in indicator pigment:chl-a ratios caused by environmental variability. Combined with flow cytometric cell counts, depth-dependent trends in the intracellular concentration and composition of phytoplankton pigments can be identified. These data show an exponential decrease in the ratio of carbon-to-chlorophyll with depth, in response to decreasing light intensity. The relative as well as absolute concentrations of phytoplankton pigments are also seen to change with depth, particularly under stratified conditions, with the ratio of zeaxanthin to chlorophyll-a decreasing with increasing depth, and the ratio of chlorophyll-b to chlorophyll-a increasing. Cluster analysis is used to identify the main phytoplankton populations in the North West Atlantic, with communities of large, fucoxanthin-containing phytoplankton dominating in spring when mixing is strong, before being replaced by smaller cells upon the onset of stratification. The links between trends in phytoplankton photophysiology and abiotic conditions are also explored, with temperature being found to be the most important forcing factor. Size-class specific relationships between phytoplankton photosynthetic rates and temperature are identified, with the potential for use in remotely-sensed models of primary production.

M. Tech (Biosciences, Faculty of Applied and Computer Sciences), Vaal University of Technology. / The mass production of algae for commercial purposes has predominately been carried out in open ponds systems. However, open ponds systems have a number of disadvantages such as poor light utilization, requirement for large areas of land and high risks of contamination. On the other hand, photobioreactors have attracted much interest because they allow a better control of the cultivation conditions than open systems. With photobioreactors, higher biomass productivities are obtained and contamination can be easily prevented. Photobioreactors can also be engineered to manipulate the light and dark photosynthetic reactions thus enhancing biomass productivity. The main objective of this study was to construct a novel tubular photobioreactor which had the ability to expose the cultured alga to light and dark phases with the aim of optimizing the algal biomass production. A novel tubular photobioreactor with the ability to manipulate the cultured alga’s light and dark photosynthetic reactions was constructed in this study. The alga Spirulina platensis was chosen as the test organism in this novel tubular photobioreactor due to a number of reasons such as its globally socioeconomic importance, its tolerance of higher pH and temperature values which makes it almost impossible to contaminate. The cultivation process of Spirulina in the photobioreactor was investigated through alternating light and dark cycles in an attempt to increase the photosynthetic efficiency of the culture. The effect of different light intensities on the growth of Spirulina in the novel tubular photobioreactor was investigated and it was found that the best light condition that favored higher biomass formation was at 600 μ mol m-2 s-1. Five different light/ dark ratios were evaluated at a light intensity of 600 μ mol m-2 s-1 during a batch mode of operation of the novel tubular photobioreactor. The light/ dark ratio of 1:0.25 was found to be the best ratio because it gave the highest biomass in the shortest period of time when compared to the other ratios used. These results seem to suggest that longer light cycle relative to dark cycle results in higher biomass production. The ratio of 1:0.25 was then used to operate the novel tubular photobioreactor in a continuous mode. A maximum biomass productivity of 25 g/m2/day was achieved which corresponded to a net photosynthetic efficiency of 5.7 %. This result was found to be higher than what most photobioreactors could achieve but it was 2.8 g/m2/day lower than the highest ever reported productivity in a photobioreactor when Spirulina is cultivated. The 2.8 g/m2/day lower was attributed to the different materials used in the construction of these two photobioreactors. The photobioreactor which achieved 27.8 g/m2/day was made up of a clear glass whereas the novel tubular photobioreactor was made up of a PVC tubing. PVC tubes tend to change from clear to a milky colour after a certain period when it is used at higher temperature and pH values hence blocks a certain amount of light. Therefore the main recommendation in this study is to use a PVC tubing with a longer life span when used at a higher temperature and pH values.

Les cyanobactéries des genres Microcystis et Planktothrix sont des micro-organismes capables de proliférer dans de nombreux plans d'eau douce. Ces proliférations sont associées à des risques pour la santé humaine et animale en raison de la production de produits naturels, parmi lesquels des toxines. Ces molécules présentent une diversité de structure chimique de d'activités biologiques d'intérêt pour l'industrie pharmaceutique et biotechnologique. Nous avons revisité le potentiel en produits naturels de Microcystis et Planktothrix. Par des approches complémentaires de biologie moléculaire, génomique et transcriptomique, nous avons caractérisé des gènes impliqués dans la synthèse de ces produits naturels ainsi qu'exploré leur évolution et la régulation de leur expression. Ces travaux ont permis de mettre à jour une distribution, des évènements évolutifs et des profils d'expression surprenants et permettent d'envisager de nouvelles applications pour les produits naturels. / Microcystis and Planktothrix are cyanobacterial genus commonly proliferating in freshwater ecosystems. These blooms are associated with human and animal health threat because of synthesis of natural products and cyanotoxins. These compounds are of great chemical diversity and of interest for biotechnological and pharmaceutical applications. We revisited the natural products potential of Microcystis and Planktothrix. Combining molecular biology, genomics and transcriptomics investigations, we characterized natural products gene clusters. We studied their distribution, evolution and transcription as well. This work uncovered new distribution pattern, evolutionary events and unexpected expression patterns. These insights will allow new investigations and applications for cyanobacterial natural products.

Marine productivity along the western Antarctic Peninsula (WAP) is declining. The WAP is site of the fastest regional warming in the southern hemisphere, and has experienced atmospheric and oceanic temperature increases leading to increased glacial inputs and reduced winter sea-ice cover. Sea-ice is a key link between climate and phytoplankton production, as melting sea-ice stratifies the water column and provides a source of micronutrients to surface waters. Reductions in ice cover have been accompanied by declining chlorophyll (chl; a proxy for phytoplankton biomass), and a shift to smaller cell sizes in phytoplankton communities. These reductions have implications for carbon drawdown and production available to higher trophic levels. However, little is known about phytoplankton shifts at the community level, as existing studies are based on satellite records and photosynthetic pigment analyses. To elucidate the nature of the changes within phytoplankton assemblages, high-resolution time-series data of diatom speciation are coupled to environmental data from five years in Ryder Bay (Adelaide Island, WAP). Long-term monitoring at this site by the British Antarctic Survey has identified a strong relationship between chl and water column stratification, and this study spans a wide range of physical conditions and biological production. By comparing high- and low-chl phytoplankton assemblages, this study investigates the mechanisms underlying productivity changes and the manner in which these changes impact nutrient cycling, drawdown and trophic transfer. The results presented here are the first full season in-situ records documenting differences in phytoplankton and diatom assemblages between highand low-chl years. The primary difference between chl conditions is a dramatic decline in diatom abundance. This analysis indicates that the mechanism producing low-chl seasons is less stratified surface waters, where light levels are much more variable than in high-chl years. Overall production is reduced, and small increases are seen in biomass of prymnesiophytes, which are better adapted to variable light. These shifts in phytoplankton composition and size structure are consistent with a southward propagation of observed climate change effects. Within the diatom community, changes in seasonal succession and a decrease in species richness occurred following low winter sea-ice. As the main component of high productivity and that most efficiently transferred to higher trophic levels, variation in diatom production due to environmental conditions is a mechanism to explain the observed WAP ecosystem changes and chl decline. Changes in phytoplankton stocks and composition also affect nutrient use, and here the use of silicon and iron (Si and Fe, respectively, which limit productivity in large areas of the Southern Ocean) is investigated. Seasonal Si budgets estimated from Si isotopes indicate a 40 – 70% decline in Si use between high-chl and intermediate-chl years, in agreement with other indices of productivity. The consequences of reduced demand and changing supply suggest future accumulation of Si in WAP surface waters. This should increase Si export away from the WAP shelf, which may act as a mechanism to enhance productivity and carbon drawdown in the wider Southern Ocean. Sources of Fe were assessed by direct measurement and naturally occurring radioisotopes of radium. These reveal significant inputs at the surface (due to glacial sources) and to deep waters (from shelf/slope sediments), which dominate supply to the surface mixed layer at different times. Iron availability and nutrient drawdown indicate that Fe is supplied to WAP surface waters in excess of biological demand. Projected changes to Fe sources and sinks indicate that continued warming will increase the WAP Fe inventory. As for Si, this excess Fe may also be advected away from the shelf, making this region a net Fe source to the Southern Ocean.

Les diatomées sont des micro-algues largement utilisées pour évaluer la qualité écologique des milieux aquatiques. La grande majorité des indices biotiques utilisant les diatomées sont basés sur la sensibilité à la pollution des espèces. Cela constitue un frein à leur utilisation car l’identification taxonomique au niveau de l’espèce est complexe, longue, coûteuse et source d’erreurs. Afin de rendre le processus d’identification plus simple, des indices biotiques basés sur des niveaux taxonomiques supérieurs à l’espèce, comme le genre, ont été mis au point. Mais la perte d’informations associée à la réduction de la résolution taxonomique est susceptible de rendre ces outils moins efficaces.Une approche alternative et plus récente propose de baser la simplification, non pas sur la taxonomie, mais sur la phylogénie. Cette approche fait implicitement l’hypothèse qu’il existe un signal phylogénétique dans les préférences écologiques des espèces, c’est à dire que deux espèces phylogénétiquement proches sont davantage susceptibles de présenter des réponses écologiques similaires que deux espèces prises au hasard. Si un tel signal existe, il implique une possible redondance phylogénétique dans les outils de bioindication existants, en particuliers ceux basés sur les niveaux taxonomiques les plus fins. L’objectif est de mettre à profit ce signal pour simplifier l’évaluation écologique des milieux aquatiques.Ce travail s’attache à développer cette approche chez les diatomées et se décompose en trois parties. Nous présentons d’abord un nouveau package R entièrement dédié à l’analyse du signal phylogénétique et à l’étude de la distribution des valeurs de traits dans les phylogénies. Nous démontrons ensuite la présence d’un signal phylogénétique pour de nombreux traits écologiques chez les diatomées d’eau douce. Ces traits sont les optimums écologiques de 127 espèces pour un ensemble de paramètres physico-chimiques, mesurés pendant huit ans dans des cours d’eau de l’est de la France. Nous montrons que le signal est variable en fonction des traits mais que la niche écologique des espèces étudiées est, de manière générale, dépendante de la phylogénie. Dans une troisième partie, nous proposons une méthode pour extraire des clusters d’espèces partageant des traits similaires tout en étant phylogénétiquement proches. Nous mettons en œuvre cette méthode sur des données de sensibilités aux pollutions pour démontrer les possibilités de simplification des indices biotiques basés sur les diatomées en prenant en compte la redondance phylogénétique. Nos résultats tendent à montrer que le potentiel de simplification en utilisant la phylogénie comme guide est significatif. / Diatoms are micro-algae commonly used to assess the ecological quality of freshwaters. Most of the biological indices using diatoms are based on species sensitivity to pollutions. This constitutes an obstacle to the use of diatoms in ecological assessment because taxonomical identification at species level is difficult, time consuming, costly and source of errors. To avoid this problem, scientists developed biological indices based on higher taxonomical levels like the genus. However, the loss of information caused by the taxonomical resolution decrease can make these methods less efficient.A more recent alternative proposes to use the phylogeny to simplify ecological assessment methods. This approach makes the implicit hypothesis that there is a phylogenetic signal for species ecological preferences, i.e. that closely related species are more likely to share similar ecological preferences than species taken randomly. If such a signal exists, it may mean that there is a phylogenetic redundancy in bioassessment tools, especially the ones which are based at species level. The aim is to exploit this signal to simplify the biological assessment of aquatic ecosystems.This work aims to develop this approach with diatoms and is divided in three parts. First, we introduce a new R package dedicated to the analysis of the phylogenetic signal and to study traits values patterns in phylogenies. In a second part, we demonstrate the presence of phylogenetic signal for many ecological traits in freshwater diatoms. These traits are the ecological optima of 127 species for a set of physical and chemical parameters. They were estimated from data collected during 8 years in rivers in eastern France. We show that the strength of the signal varied significantly from one trait to another but, overall, diatoms ecological niches are related to the phylogeny. Finally, in a third part, we introduce a new method to extract clusters of species sharing similar traits and being phylogenetically related. We apply this method on pollutions sensitivities data in order to demonstrate the possibility to simplify biological indices based on diatoms by taking account of phylogenetic redundancy. Our results suggest that phylogenetic approaches offer a scope for simplification without an important loss of ecological information.

Recent observations have shown that many sandstone buildings, including important components of the UK's cultural heritage, are becoming covered with green algal growths. This is likely to result from recent changes in air quality and the impacts of a changing climate. The northern regions of the UK in particular have an abundance of sandstone heritage and, given the likelihood of warmer, wetter winters here, algal growth on vulnerable monuments is likely to become a primary conservation concern over the next 50 years. Observations of sandstone monuments in the northern regions of the UK, in particular in Belfast (Northern Ireland), Sheffield and Edinburgh have highlighted that algal greening is notably patchy. This is likely due to the array of factors which affect the bioreceptivity of host substrates such as sandstone. The bioreceptivity of a substrate (its ability to become colonised by microbes such as green algae) is dependent on inherent, external and architectural factors. The role of these factors and the interrelationships between them requires further study. This thesis aims to investigate the inherent, external and architectural factors which encourage colonisation of sandstone by green algae through an integrated programme of laboratory and field experimentation. The primary objectives of this study are: to develop improved laboratory experimental methods to control and monitor algal growth, to investigate the role of external, inherent and architectural factors and to explore the fundamental role of moisture in the development of algal greening. In order to address these objectives, laboratory and field experiments have been linked within an integrated overall methodology. Short-term laboratory experiments have investigated the bioreceptivity of four British sandstones (Peak Moor, Dungannon, St Bees and 'baluster stone') to single and mixed green algal treatment with Stichococcus bacillaris, Chlorella vulgaris and Desmococcus olivaceus, under controlled conditions. Two field experiments have also been conducted. The first exposed unweathered blocks of Dungannon sandstone in the wet environment of Derrygonnelly, Northern Ireland for 30 months. The second exposed reclaimed sandstone balusters in a shaded and exposed site in central Oxford for 12 months. The laboratory and field experiments presented utlilise a range of simple and accessible methods to monitor biofilm development (for example novel methods to map biomass) and changes in substrate condition (such as monitoring surface moisture movements with weight change and hand-held moisture meters, and using light microscopy to help visualise the impact of green algal biofilms). The results presented in this thesis confirm that moisture plays a fundamental role in the development of green algal biofilms. In laboratory experiments, colonisation often occurred within a consistent moisture zone and preferential greening in field experiments was observed in areas of frequent moisture movement. External factors have been shown to have a strong influence, in laboratory experiments where marine salts were applied, these were found to delay colonisation by around seven days. Furthermore, salts resulted in inhomogeneous patterns of colonisation, similar to those observed in scoping studies conducted in Sheffield. Laboratory experiments have also demonstrated that inherent substrate factors such as high porosity and presence of certain minerals (such as clay laminations in Dungannon) can increase the primary bioreceptivity of sandstone surfaces. Field experiments have demonstrated that architectural factors such as aspect and geometry can increase the bioreceptivity of exposed samples. In particular, preferential greening was observed on the dynamically wetted south west facing blocks in Derrygonnelly and on exposed compared with shaded balusters in Oxford. Greening was also concentrated in areas of rainwater flows and stores. Investigation of the role of external, inherent and architectural factors in the development of algal greening as provided by this project, supplies useful information for those managing our sandstone cultural heritage. This will enable more informed decisions to be made over appropriate management and conservation strategies for the future.

In my thesis I present data collected from a long-term selection experiment using the freshwater model organism Chlamydomonas reinhardtii. The selection experiment was designed to disentangle the effects of the number of multiple environmental drivers (MEDs) and the identity of those environmental drivers including high CO2, high temperature, general nutrient depletion, reduced light intensity, reduced phosphate availability, the addition of a herbicide, UV radiation and reduced pH. Using up to eight environmental drivers, I show how simple organisms such as C. reinhardtii evolve in response to MEDs. The first step in this investigation is to examine the short-term response of MEDs. Data collected at the beginning of the selection experiment will provide insight into the early stages of microevolution by investigating key differences in the short-term (plastic) responses to few vs. many MEDs. Here, I focus on how the data collected from the responses to single environmental drivers can help us predict the responses to MEDs by using ecological models (additive, comparative, multiplicative). I show that the short-term plastic responses to single environmental drivers can predict the effect of MEDs using the comparative model because the response is largely driven by the single dominant driver present. I also demonstrate the importance of the number of environmental drivers (NED) for making predictions from the single environmental drivers and show that predictions become more reliable as the NED increases. The results gathered from short-term responses provide evidence that single environmental driver studies are useful for predicting the effect of MEDs. After evolution, I found that the strength of selection varies with NED in a predictable way, which connects the NED to the evolutionary response (size of the direct response) through the strength of selection. Here, I used statistical models to quantify the effect of NED on the evolutionary response to MEDs and then interpreted this by considering the possible genetic constraints on adaptation to MEDs. A subset of populations evolved in environments with five environmental drivers and all populations evolved in the single environmental driver environments are used to examine how adapting to single vs. many environmental drivers affect local adaptation. I examine how populations selected in environments with one environmental driver, five environmental drivers and the evolved control, differ in their response to new environments with the same NED, environments with different NED, and a novel environment. I found that there is a relationship between local adaptation and the strength of selection in the local environment and patterns of local adaptation are affected by the NED of new environments. Lastly, I present the phenotypic consequences of evolution under MEDs. I found that before evolution, measures of chlorophyll content and cell size decline with increasing NED. However, after evolution the relationship between chlorophyll content and cell size with NED is weaker because populations converge on the same phenotypes as they evolve. I also present a case-study of how mass spectrometry methods can be used to better understand underlying molecular mechanisms of two phenotypes (chlorophyll positive and chlorophyll negative cells). This selection experiment is a good example of how laboratory investigations and model organisms can be used to design experiments with enough replication to have high statistical power in order to make more accurate predictions on the short- long-term effects of MEDs. Whilst there have been some studies on the effects of MEDs, these studies rarely have more than three environmental drivers (sometimes 5 environmental drivers) and there are only a handful of long-term MED studies. This study can be used to develop a priori hypotheses for investigating how environmental change will shape natural microbial communities, and is especially useful for organisms where long-term studies with multiple environmental drivers are unfeasible.