Growth and release of extracellular organic compounds by benthic diatoms depend on interactions with bacteria

Phototrophic epilithic biofilms harbour a distinct assemblage of heterotrophic bacteria, cyanobacteria and photoautotrophic algae. Secretion of extracellular polymeric substances (EPS) by these organisms and the physicochemical properties of the EPS are important factors for the development of the biofilms. We have isolated representative diatom and bacteria strains from epilithic biofilms of Lake Constance. By pairwise co-cultivating these strains we found that diatom growth and EPS secretion by diatoms may depend on the presence of individual bacteria. Similar results were obtained after addition of spent bacterial medium to diatom cultures, suggesting that soluble substances from bacteria have an impact on diatom physiology. While searching for putative bacterial signal substances, we found that concentrations of various dissolved free amino acids (DFAA) within the diatom cultures changed drastically during co-cultivation with bacteria. Further, the secretion of extracellular carbohydrates and proteins can be influenced by bacteria or their extracellular substances. We have performed mass spectrometric peptide mapping to identify proteins which are secreted when co-cultivating the diatom Phaeodactylum tricornutum Bohlin and Escherichia coli. The identified proteins are possibly involved in signalling, extracellular carbohydrate modification and uptake, protein and amino acid modification, and cell/cell aggregation of diatom and bacteria strains. Our data indicate that diatom-bacteria biofilms might be regulated by a complex network of chemical factors involving EPS, amino acid monomers and other substances. Thus interactions with bacteria can be considered as one of the main factors driving biofilm formation by benthic diatoms.     

Structure and Dynamics of Exopolymers in an Intertidal Diatom Biofilm

Diatom biofilms growing at the surface of the intertidal mudflat of Marennes Oléron, France, were incubated for 48 h in the laboratory under simulated conditions of high- and low tide (immersed and emersed in seawater) and day and night (illuminated or dark conditions). The biofilms were subsequently sampled using the cryolander technique, without disturbing the structure. The samples were kept in liquid nitrogen until they were transferred to the cooled stage of a field-emission cryo-scanning electron microscope, which was used to study the structural relationships between the sediment particles, the diatoms and the different types of extracellular polymeric substances (EPS) produced by these organisms. The diatoms were most abundant at the sediment surface when incubated in the light under emersed conditions. In the dark or when immersed, the diatoms migrated into the sediment. In the light, the diatoms were coated with EPS, while this was not the case when incubated in the dark. When immersed, the sediment surface appeared smooth as the result of the deposition of mud. Under emersed conditions, the coarser silt grains were prominently present. These grains were wrapped with organic matter and bound together through threads of EPS. This was the case both in light and in dark incubated sediment. It is proposed that this latter type of EPS contributes to the increased erosion threshold of intertidal mudflats colonized by biofilms of diatoms.     

Utilizing QCM-D to characterize the adhesive mucilage secreted by two marine diatom species in-situ and in real-time

The quartz crystal microbalance with dissipation monitoring (QCM-D) was used to monitor the deposition of adhesive extracellular polymeric substances (EPS) employed by the marine biofouling diatoms Craspedostauros australis Cox and Amphora coffeaeformis Cleve during initial adhesion and subsequent motility. Upon injection into the QCM chamber, initial negative frequency (f) shifts and positive dissipation (D) shifts were measured that correlated to cells impacting and adhering to the QCM sensor surface. Following this "initial adhesion" response, f continued to decrease while D increased logarithmically. Rather than the result of any cell morphological alterations at the substrate surface, the shifts were correlated to the time-dependent deposition of EPS upon the substrate surface as a result of cellular motility, or gliding. Experiments utilizing comparable cell concentrations of the diatom species C. australis and A. coffeaeformis revealed significant differences between the parameter responses recorded, where A. coffeaeformis produced Deltaf and DeltaD values of -32 Hz and 6.6, and C. australis produced values of -82 Hz and 42, respectively, after 20 h post-inoculation. The viscoelastic properties of the adhered EPS adlayer from both species were examined via a Deltaf/DeltaD plot, providing reproducible signature "ratio" values for each species that likely correlate to differences in EPS interactions with the substrate that may be associated directly to differences in the fouling potential of the two species. There is a distinct lack of knowledge regarding the chemical nature of the adhesive polymers engaged, and few quantitative techniques are applicable to the study of diatom EPS. We propose that QCM-D may be a useful tool in identifying differences in the EPS employed by diatoms of different fouling potential.     

Biogenic Stabilization of Intertidal Sediments: The Importance of Extracellular Polymeric Substances Produced by Benthic Diatoms

The sediment-stabilizing effect of benthic diatoms was investigated in a laboratory setting. Axenic cultures of the benthic diatoms Nitzschia cf. brevissima and Cylindrotheca closterium were inoculated in Petri dishes containing sand and incubated under axenic conditions. By ensuring aseptic routines throughout the experiments, interference from other organisms occurring with diatoms in natural photothrophic biofilms was avoided. This allowed the examination of the role of benthic diatoms in sediment stabilization. Increases in the critical erosion shear stress of the sediment were observed in the presence of both diatom taxa relative to sterile sediment. However, N. cf. brevissima was more effective than C. closterium. Values of critical shear stress in the experimental system were in the same range as those observed in natural biofilms, which indicates that diatoms are important agents for biogenic stabilization. Extracellular carbohydrate contents in the microcosms were similar for both diatom species. However, in the presence of N cf. brevissima, extracellular carbohydrate correlated significantly to critical shear stress, explaining up to 80% of the variation, whereas this was not the case for C. closterium. Therefore, it was concluded that the quantity of extracellular polymeric substances (EPS) alone did not explain the biogenic stabilization. Observed adsorption of EPS to sediment particles depended on the relative amount of uronic acids in the exopolymers. Using fluorescently labeled lectins, confocal laser scanning microscopy showed that EPS secretion by N. cf. brevissima resulted in ordered three-dimensional matrix structures. It is suggested that the structuring of EPS plays an prominent role in the process of biostabilization, and that diatoms such as N. cf. brevissima are actively involved in producing the structure of EPS, whereas others such as C. closterium do not do so to the same extent.     

Assembly rules and community models for unicellular organisms: patterns in diatoms of boreal streams

1. Many studies have addressed either community models (e.g. Clementsian versus Gleasonian gradients) or assembly rules (e.g. nestedness, checkerboards) for higher plant and animal communities, but very few studies have examined different non‐random distribution patterns simultaneously with the same data set. Even fewer studies have addressed generalities in the distribution patterns of unicellular organisms, such as diatoms. 2. We studied non‐randomness in the spatial distribution and community composition of stream diatoms. Our data consisted of diatom surveys from 47 boreal headwater streams and small rivers in northern Finland. Our analytical approaches included ordinations, cluster analysis, null model analyses, and associated randomisation tests. 3. Stream diatom communities did not follow discrete Clementsian community types, where multiple species occur exclusively in a single community type. Rather, diatom species showed rather individualistic responses, leading to continuous Gleasonian variability in community composition. 4. Although continuous variability was the dominating pattern in the data, diatoms also showed significant nestedness and less overlap in species distribution than expected by chance. However, these patterns were probably only secondary signals from species’ individualistic responses to the environment. 5. Although unicellular organisms, such as diatoms, differ from multicellular organisms in several biological characteristics, they nevertheless appear to show largely similar non‐random distribution patterns previously found for higher plants and metazoans.     

Microphytobenthos,their Extracellular Polymeric Substances,and the Morphogenesis of Intertidal Sediments

Intertidal sediments are important areas that separate the land from the sea and form natural coastal defenses. They are known as highly productive ecosystems, fueling the coastal food web. It is also conceived that microphytobenthos contribute to the stability of intertidal sediments by increasing the erosion threshold and that they are major players in coastal morphodynamics. Depending on the sedimentary composition of intertidal flats, different types of microphytobenthos colonize the sediment surface. Fine sand sediment is often colonized by cyanobacteria, prokaryotic algae, which form dense and rigid microbial mats. Mudflats on the other hand are characterized by the development of thin biofilms of epipelic diatoms. Both groups of phototrophic microorganisms excrete extracellular polymeric substances (EPS), but they do so in different ways and for different reasons. Two operationally defined fractions, water- and EDTA-extractable EPS, have been obtained from intertidal diatom biofilms and from cultures. They differ in composition and their production seems to be under different metabolic control. Water-extractable EPS are considered to be closely associated with the diatoms and are rich in neutral sugars, notably glucose. These EPS show a dynamic relationship with the microphytobenthic biomass. EDTA-extractable EPS are tightly bound to the sediment, probably through bridging by divalent ions. This material is rich in uronic acids and other acid sugars and is weakly related to chlorophyll. These EPS have been conceived to be a major factor in the structuring and diagenesis of coastal sediments and essential for increasing the sediment erosion threshold. However, this relationship is now questioned.     

Biofilm and capsule formation of the diatom Achnanthidium minutissimum are affected by a bacterium

Photoautotrophic biofilms play an important role in various aquatic habitats and are composed of prokaryotic and/or eukaryotic organisms embedded in extracellular polymeric substances (EPS). We have isolated diatoms as well as bacteria from freshwater biofilms to study organismal interactions between representative isolates. We found that bacteria have a strong impact on the biofilm formation of the pennate diatom Achnanthidium minutissimum. This alga produces extracellular capsules of insoluble EPS, mostly carbohydrates (CHO), only in the presence of bacteria (xenic culture). The EPS themselves also have a strong impact on the aggregation and attachment of the algae. In the absence of bacteria (axenic culture), A. minutissimum did not form capsules and the cells grew completely suspended. Fractionation and quantification of CHO revealed that the diatom in axenic culture produces large amounts of soluble CHO, whereas in the xenic culture mainly insoluble CHO were detected. For investigation of biofilm formation by A. minutissimum, a bioassay was established using a diatom satellite Bacteroidetes bacterium that had been shown to induce capsule formation of A. minutissimum. Interestingly, capsule and biofilm induction can be achieved by addition of bacterial spent medium, indicating that soluble hydrophobic molecules produced by the bacterium may mediate the diatom/bacteria interaction. With the designed bioassay, a reliable tool is now available to study the chemical interactions between diatoms and bacteria with consequences for biofilm formation.     

THE COMPLEX EXTRACELLULAR POLYSACCHARIDES OF MAINLY CHAIN‐FORMING FRESHWATER DIATOM SPECIES FROM EPILITHIC BIOFILMS1

Diatoms are dominant organisms in phototrophic biofilms in aquatic habitats. They produce copious amounts of extracellular polymeric substances (EPS), which mainly consist of carbohydrates and traces of proteins and glycoproteins. This study focuses on the characterization of EPS from a total of 14 diatoms belonging to the six genera Achnanthes, Cymbella, Fragilaria, Punctastriata, Staurosira, and Pseudostaurosira, all of which were isolated from epilithic biofilms of the littoral zone of Lake Constance. EPS from all isolates were extracted by a sequential extraction procedure resulting in five different fractions. The monosaccharide composition of each fraction was analyzed by HPLC equipped with a pulse amperiometric detector, yielding results similar to those obtained by probing the EPS structures with monomer‐specific fluorophore‐linked lectins. Significant differences in carbohydrate composition occurred in the different fractions of single isolates. Most of the diatom isolates in our study form chain‐like colonies in which the cells are attached to each other by intercellular pads. Here we demonstrate that these pads can be dissolved in hot bicarbonate and that they show a heterogeneous composition of monosaccharides in contrast to other fractions, which mostly were dominated by one or two monosaccharides. Principal component analysis indicates a correlation between carbohydrate composition of EPS fractions and the phylogenetic relationship of the respective species, indicating that EPS analyses under defined culture conditions may support taxonomic analyses.     

Effect of chlorination on the development of marine biofilms dominated by diatoms

This study addressed the antifouling efficiency of commercially available chlorine at different concentrations (0.5%, 1%, and 2%) and exposure times (0.5?min, 1?min, 5?min, and 15 min). The rapid and non-destructive FIRe (fluorescence induction and relaxation) technique was used to evaluate the effects of the biocide on diatom dominated biofilms. The efficiency of chlorine in removing diatoms from the developed biofilms increased with an increase in concentration and exposure time. The fluorescence measurements revealed low F(v)/F(m) and high σ(PSII) values for chlorine-treated Navicula and Amphora biofilms indicating that chlorination was efficient in damaging the photosystem-II reaction centers. Chlorination also caused mortality of diatom cells by damaging the cell body. In natural biofilms, the biocidal effect of chlorine was species specific; species of Amphiphrora, Navicula, Cylindrotheca, and Coscinodiscus showed an increase in the density of the population, but species of Pleurosigma, Amphora, and Thalassionema did not increase in density after chlorine treatment. It was also demonstrated that diatoms can colonize, grow and photosynthesize on chlorine-treated surfaces. Under pulse chlorination (treatment every 6 h), irrespective of chlorine concentration, the development of biofouling decreased with an increase in exposure time. Differences between exposure times of 1 to 15?min were not significant. Additionally, transmission levels of the control (non-chlorine-treated) fouled coupons reduced significantly (～20%) compared to the chlorine-treated fouled coupons (<2%). These results suggest that chlorine can be used as a biocide to control the development of diatom biofilms.     

Dynamics of diatom colonization process in some rivers influenced by urban pollution (Hanoi, Vietnam)

Periphytic diatom communities’ colonization patterns were studied at three sampling stations of the Red–Nhue–Tolich hydrosystem presenting different urban pollution levels by using artificial substrates for 6 weeks in dry season 2005. Structural characteristics of periphytic diatoms developed on glass substrates at each sampling site were followed and compared. This experiment showed, through various general criteria (total diatom density, dry weight biomass) and specific criteria (relative diatom abundances, indices) that the structure of benthic diatoms developed on these substrates was strongly affected by pollution as early as the second week of colonization. Communities exposed to heavily and moderately polluted sites contained species which are known to be saprophilous or tolerant to organic pollution such as Nitzschia umbonata, Nitzschia palea, Cyclotella meneghiniana, Eolimna minima. Growth inhibition of diatom communities at the heavily polluted site was mostly related to a strong increase of organic load rather than to low metallic input, though metallic (Cd and Zn) burdens have been successfully quantified in the biofilms developed at the three studied sites. Nevertheless, no significant difference in species richness and diversity index between colonization duration times was observed. Based on values of diatom indices Indice de Polluosensibilité (IPS) and diatom assemblage index to organic water pollution (DAIPo), water quality could be classified as medium quality at Red site, polluted at NT₂ and heavily polluted at TL. Thus, the use of diatoms as a tool for water assessment appears suitable for monitoring rivers in Vietnam, as it is in several other countries.     

Changes in diatom-dominated biofilms during simulated improvements in water quality: implications for diatom-based monitoring in rivers

Although benthic diatoms are used to assess river water quality, there are few data on the rate at which diatom assemblages react to changes in water quality. The aim of this study was to assess the reaction time of diatoms and to discuss the changes occurring during water quality improvement on the basis of their autecological characteristics. In order to simulate this improvement, diatom-dominated biofilms grown on artificial sandstone substrata were transferred from several polluted rivers to an unpolluted river. They were sampled three times: before transfer and 1 and 2 months after transfer. The ecology and growth-forms of the taxa explained most of the changes in species composition observed during the experiment. Adnate diatoms gradually replaced motile and stalked taxa. Gomphonema parvulum, a stalked diatom positioned vertically in the biofilm, is adapted for light and space competition in high-density algal biofilms. When transferred to an unpolluted site, this growth-form is less competitive and does not tolerate the high grazing pressure. Fistulifera saprophila is a single celled motile diatom, living in organic matrices. When the artificial substrata were transferred to the unpolluted site, this particular ecological niche disappeared quickly. On the other hand, Achnanthidium minutissimum, which is considered to be cosmopolitan and an early colonizer, increased during the first month of transfer and then decreased. It was gradually replaced by A. biasolettianum, which was the taxon best suited to this pristine stream. The changes observed differed between treatments depending on the species composition and architecture of the biofilms. In particular, biofilms dominated by stalked and motile diatoms were more quickly modified than those dominated by small motile diatoms. The diatom index reflects these changes, and its values showed that about 60 days following a water quality improvement were necessary for transferred diatom assemblages to reach diatom index values similar as those at the unpolluted river.     

Correlation of surface sediment diatoms with the present lake water pH in 28 Algoma lakes,Ontario, Canada

The surface sediment diatom analysis of 28 Algoma lakes (pH 4.40–8.13) indicates that even though each lake has a widely different aquatic environment and characteristic diatom assemblage, a definite relationship exists between the lake water pH and their diatom assemblages. In the acidic lakes acidobiontic and acidophilous diatom species predominate whereas in circumneutral and alkaline lakes circumneutral and alkaliphilous diatoms were most common. Cluster analysis of the pH indicator diatom assemblages grouped the study lakes into three distinct cluster groups. These groups also closely corresponded to lake water pH. On the basis of published ecological information as well as their presence in our study lakes, the pH indicator status of a number of diatom taxa have been discussed. A detailed listing of the diatom taxa identified and their pH indicator status is provided in order to facilitate their use in future diatom-inferred pH studies.     

The development of marine biofilms on two commercial non-biocidal coatings: a comparison between silicone and fluoropolymer technologies

The antimicrobial performance of two fouling-release coating systems, Intersleek 700? (IS700; silicone technology), Intersleek 900? (IS900; fluoropolymer technology) and a tie coat (TC, control surface) was investigated in a short term (10 days) field experiment conducted at a depth of ca 0.5 m in the Marina Bandar Rawdha (Muscat, Oman). Microfouling on coated glass slides was analyzed using epifluorescence microscopy and adenosine-5'-triphosphate (ATP) luminometry. All the coatings developed biofilms composed of heterotrophic bacteria, cyanobacteria, seven species of diatoms (2 species of Navicula, Cylindrotheca sp., Nitzschia sp., Amphora sp., Diploneis sp., and Bacillaria sp.) and algal spores (Ulva sp.). IS900 had significantly thinner biofilms with fewer diatom species, no algal spores and the least number of bacteria in comparison with IS700 and the TC. The ATP readings did not correspond to the numbers of bacteria and diatoms in the biofilms. The density of diatoms was negatively correlated with the density of the bacteria in biofilms on the IS900 coating, and, conversely, diatom density was positively correlated in biofilms on the TC. The higher antifouling efficacy of IS900 over IS700 may lead to lower roughness and thus lower fuel consumption for those vessels that utilise the IS900 fouling-release coating.     

Impact of elevated UV-B radiation on photosynthetic electron transport, primary productivity and carbon allocation in estuarine epipelic diatoms

Epipelic diatoms are important components of microphytobenthic biofilms. Cultures of four diatom species (Amphora coffeaeformis, Cylindrotheca closterium, Navicula perminuta and Nitzschia epithemioides) and assemblages of mixed diatom species collected from an estuary were exposed to elevated levels of ultraviolet-B (UV-B) radiation. Short exposures to UV-B resulted in decreases in photosystem II (PSII) photochemistry, photosynthetic electron transport, photosynthetic carbon assimilation and changes in the pattern of allocation of assimilated carbon into soluble colloidal, extracellular polysaccharides (EPS) and glucan pools. The magnitude of the effects of the UV-B treatments varied between species and was also dependent upon the photosynthetically active photon flux density (PPFD) to which the cells were also exposed, with effects being greater at lower light levels. Both increases in nonphotochemical quenching of excitation energy in the pigment antennae and photodamage to the D1 reaction centres contributed to decreases in PSII photochemistry. All species demonstrated a rapid ability to recover from perturbations of PSII photochemistry, with some species recovering during the UV-B exposure period. Some of the perturbations induced in carbon metabolism were independent of effects on PSII photochemistry and photosynthetic electron transport. Elevated UV-B can significantly inhibit photosynthetic performance, and modify carbon metabolism in epipelic diatoms. However, the ecological effects of UV-B at the community level are difficult to predict as large variations occur between species.     

Quantification of diatoms in biofilms: standardisation of methods

Benthic diatoms, which often dominate marine biofilms are mostly pennate along with a few centric species that have an attached mode of life. Even though the range of diatoms in biofilms is diverse, their ecology is poorly understood because of the difficulty in sampling and enumeration. Scraping or brushing are the traditional methods used for removal of diatoms from biofilms developed on solid substrata. The method of removal is the most critical step in enumerating the biofilm diatom community structure. In this study, a nylon brush and ceramic scraper were used as tools for the removal of diatoms from 1 - 4-day-old biofilms developed on fibreglass coupons and glass microscope slides. Standardisation of methods showed that the sample volume used in the analyses had the least influence on the quantification, whereas the method of removal was critical. The nylon brush was more efficient at recovering diatoms compared to a ceramic scraper. Direct microscopic enumeration of the community in the case of glass slides indicated that scraping resulted in between 30-50% underestimation. Heterogeneity in diatom community structure between replicate samples is one possible reason for such underestimation.     

Quantification of diatoms in biofilms: Standardisation of methods

Abstract

Benthic diatoms, which often dominate marine biofilms are mostly pennate along with a few centric species that have an attached mode of life. Even though the range of diatoms in biofilms is diverse, their ecology is poorly understood because of the difficulty in sampling and enumeration. Scraping or brushing are the traditional methods used for removal of diatoms from biofilms developed on solid substrata. The method of removal is the most critical step in enumerating the biofilm diatom community structure. In this study, a nylon brush and ceramic scraper were used as tools for the removal of diatoms from 1 – 4-day-old biofilms developed on fibreglass coupons and glass microscope slides. Standardisation of methods showed that the sample volume used in the analyses had the least influence on the quantification, whereas the method of removal was critical. The nylon brush was more efficient at recovering diatoms compared to a ceramic scraper. Direct microscopic enumeration of the community in the case of glass slides indicated that scraping resulted in between 30–50% underestimation. Heterogeneity in diatom community structure between replicate samples is one possible reason for such underestimation.     

Biofilm diatom community structure: influence of temporal and substratum variability

Diatoms, which are early autotrophic colonisers, are an important constituent of the biofouling community in the marine environment. The effects of substratum and temporal variations on the fouling diatom community structure in a monsoon-influenced tropical estuary were studied. Fibreglass and glass coupons were exposed every month for a period of 4 days and the diatom population sampled at 24 h intervals, over a period of 14 months. The planktonic diatom community structure differed from the biofilm community. Pennate diatoms dominated the biofilms whilst centric diatoms were dominant in the water column. Among the biofilm diatoms, species belonging to the genera Navicula, Amphora, Nitzschia, Pleurosigma and Thalassionema were dominant. On certain occasions, the influence of planktonic blooms was also seen on the biofilm community. A comparative study of biofilms formed on the two substrata revealed significant differences in density and diversity. However species composition was almost constant. In addition to substratum variations, the biofilm diatom community structure also showed significant seasonal variations, which were attributed to physico-chemical and biological changes in both the water and substratum. Temporal variations in the tychopelagic diatoms of the water were also observed to exert an influence on the biofilm diatom community. Variations in diatom communities may determine the functional ecosystem of the benthic environment.     

Physical characterization and diel dynamics of different fractions of extracellular polysaccharides in an axenic culture of a benthic diatom

The excretion of extracellular polymeric substances (EPS) by an axenic culture of the benthic diatom Cylindrotheca closterium was investigated. Two sequential extraction steps proved to be suficient to remove the bulk of the EPS present. Soluble EPS was recovered by a simple centrifugation step and represented a fraction that was not or was only loosely associated with diatom cells. For the extraction of bound EPS, different procedures were compared. The best results were obtained using distilled water as extraction solvent (1?h, 30?°C). The sugars that were recovered using this procedure were typically associated with aggregates of diatoms. In addition to the distinct differences in localization of the different types of EPS, their temporal dynamics differed in relation to the light–dark cycle. Soluble EPS were continuously released into the medium at a rate of 1.6?pg?cell^?1?day^?1. In contrast, the production of bound EPS was highly light-dependent. In the dark, this bound EPS rapidly disappeared, probably as the result of its utilization by the diatoms.     

Microscale imaging sheds light on species-specific strategies for photo-regulation and photo-acclimation of microphytobenthic diatoms

Intertidal microphytobenthic (MPB) biofilms are key sites for coastal primary production, predominantly by pennate diatoms exhibiting photo-regulation via non-photochemical quenching (NPQ) and vertical migration. Movement is the main photo-regulation mechanism of motile (epipelic) diatoms and because they can move from light, they show low-light acclimation features such as low NPQ levels, as compared to non-motile (epipsammic) forms. However, most comparisons of MPB species-specific photo-regulation have used low light acclimated monocultures, not mimicking environmental conditions. Here we used variable chlorophyll fluorescence imaging, fluorescent labelling in sediment cores and scanning electron microscopy to compare the movement and NPQ responses to light of four epipelic diatom species from a natural MPB biofilm. The diatoms exhibited different species-specific photo-regulation features and a large NPQ range, exceeding that reported for epipsammic diatoms. This could allow epipelic species to coexist in compacted light niches of MPB communities. We show that diatom cell orientation within MPB can be modulated by light, where diatoms oriented themselves more perpendicular to the sediment surface under high light vs. more parallel under low light, demonstrating behavioural, photo-regulatory response by varying their light absorption cross-section. This highlights the importance of considering species-specific responses and understanding cell orientation and photo-behaviour in MPB research.