Artificial Substrates Research Articles

Fan Mussel (Pinna nobilis L.) Spat Collection, Monitoring of Early Growth and Conservation Implications by Deploying Conventional Aquaculture Methodology

Pinna nobilis, endemic to the Mediterranean Sea, has been experiencing a gradual population decline over recent decades due to anthropogenic pressures on its ecosystems. However, since 2016, its populations have suffered significant reductions because of pathological issues affecting the species across all its habitats. Aquaculture techniquesto support the limited natural recruitment P. nobilis efforts is examined. Artificial substrates for larval attachments in aquaculture infrastructures promote the survival of the juveniles that is further enhanced through protected pre-growing “nursery” farming conditions. Specific spat collectors were placed in 2 cage-fish farms in SW Amvrakikos Gulf. The harvested spats from were transferred to pre-grow in trays hanged on a long line farm mussel that is acting as a protected “nursery”, avoiding predation and any human accidentally disturbance. The survival and growth of 12 juveniles P. nobilis spat (shell length 38.1 ± 9.2 mm) in captivity (31 October 2023–15 March2023) was investigated. Out of the 12 individuals collected, 3 were examined for the presence of pathogens; only 7 survived, exhibiting enhanced growth (shell length 54.3 ± 11.6 mm) after 134 days in the nursery. The results highlight the significant role of aquaculture techniques in efforts to conserve a threatened species as well as the need for the creation of a protocol to ensure the conservation of P. nobilis.

Different management strategies for artificial substrates on nitrification, microbial composition, and growth of Penaeus vannamei in a super-intensive biofloc system

The aim of this study was to test the effect of different management strategies for artificial substrates on the nitrification process, microbial composition of water and biofilm, and growth of Penaeus vannamei in a super-intensive biofloc system. The research was conducted over 60 days using experimental units of 200 L and had the following treatments: T1 – artificial substrate + biofloc + shrimp + aeration (control), T2 - artificial substrate + water + aeration, T3 - artificial substrate + water, and T4 - only artificial substrate. The experiment was divided into two phases. Phase 1 involved maintaining the artificial substrates under the management strategy for 30 days. Phase 2 involved the use of substrates from phase 1 in a shrimp nursery for 30 days. Pre colonized artificial substrates (Needlona®) were employed at a ratio of 200 % of the tank's lateral area to promote biofilm growth. In the phase 1, T1 used a stocking density of 500 shrimp m−3 (9.72 ± 0.50 g). In phase 2, shrimp weighing 0.10 ± 0.05 g were stocked at a density of 1750 shrimp m−3 and all treatments included artificial substrates from phase 1, aeration, water, and shrimp. Molasses was the carbon source organic used. In phase 2, ammonia control was observed in treatments T2, T3, and T4 from the 10th day onwards. T4 had a nitrite spike, controlled from day 14 onwards, suggesting recovery of the nitrifying bacteria community. At the end of the phase 2, T2 and T4 showed higher abundance of coccoid bacteria in the biofilm compared to T1 and T3. T4 also had more bacillus. The shrimp final weight was higher in T2 compared to the other treatments. These findings suggest that maintaining the substrate submerged in water (T3) can be considered a practical management for artificial substrates and that it does not limit the nitrification process between culture cycles. Furthermore, exposure of artificial substrates to air (T4) also did not affect the nitrification process, leading to the recovery of the bacterial community, and the proliferation of various bacterial groups.

Characterizations of angiotensin-converting enzyme-2 (ACE2) peptidase activity

Angiotensin (Ang) II (1-8) is a potent vasoconstrictor known for its role in hypertension. Angiotensin-converting enzyme (ACE2) converts Ang II (1-8) to a vasodilator Ang (1-7) by removing the carboxy-terminal Phe. ACE2 more recently gained attention as the receptor for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that caused the coronavirus disease 2019 (COVID-19) pandemic. Given the pathophysiological importance of ACE2, the present study examined the mechanism of ACE2 catalytic activity by comparing the ability of angiotensin molecules of various lengths to compete with the artificial fluorogenic substrate. The Fluorimetric SensoLyte 390 ACE2 Activity Assay uses an Mca/Dnp fluorescence resonance energy transfer peptide as the substrate. Results showed that the natural substrate Ang II (1-8) competed with the fluorogenic substrate, reducing the fluorescence signals. Deletion of C-terminal Phe resulted in the loss of the ability to compete with the artificial substrate, as shown by the actions of Ang (1-7), Ang (2-7), and Ang (5-7). By contrast, the loss of N-terminal Asp potentiated the ability to compete with the substrate as seen by the action of Ang III (2-8). However, the loss of two amino acids (Asp-Arg) from the N-terminus reduced the ability to compete with the substrate as observed by the actions of Ang IV (3-8) and Ang (5-8). Ang I (1-10) and Ang (1-9) did not strongly compete with the substrate. Interestingly, shorter peptides Ang (1-5) and Ang (1-4) potentiated the ACE2 activity. These results suggest that Ang II and Ang III are the best natural substrates for ACE2.

Production of active human iduronate-2-sulfatase (IDS) enzyme in Nicotiana benthamiana

Many strategies have been developed to produce high levels of biologically active recombinant proteins in plants for biopharmaceutical purposes. However, the production of an active form of human iduronate-2-sulfatase (hIDS) for the treatment of Hunter syndrome by enzyme replacement therapy (ERT) is challenging due to the requirement for cotranslational modification by a formylglycine-producing enzyme encoded by sulfatase modifying factor 1 (hSUMF1) at the Cys84 residue, which converts it to C(alpha)-formylglycine. In this study, we have shown that hIDS can be highly expressed in N. benthamiana by using different constructs. Among them, BiP-GB1-L-dCBD1-2L-8xHis-L-6xHis-3L-EK-hIDS-HDEL (GB1-CBD1-hIDS) showed a high expression level when transiently co-expressed with the turnip crinkle virus gene silencing suppressor P38 and GB1-fused human calreticulin (GB1-CRT1) as a folding enhancer. The hSUMF1 was co-expressed with hIDS for cotranslational modification. The full-length recombinant proteins were purified using Ni2+-NTA affinity resin followed by enterokinase treatment to obtain tag-free hIDS. The N-terminal fragment was removed using microcrystalline cellulose (MCC) beads. The purified active form of hIDS can successfully cleave the sulfate group from an artificial substrate, 4-nitrocatechol sulfate, at a similar level to commercial hIDS expressed in animal cells. These results suggest that plants could be a promising platform for the production of recombinant hIDS.

A method to detect enzymatic reactions with field effect transistor

Study of enzyme-substrate interactions is a task of great practical and scientific importance. This paper describes the application of ion-sensitive field effect transistors in quasi-equilibrium state for examination of enzymatic reactions. A reaction occurring in the liquid gate affects the chemical potential of electrons in this gate, and this phenomenon may be used to explore biochemical interactions. This strategy can be applied to detect interactions of enzymes with substrates, inhibitors and activators regardless of their optical and electrochemical properties. Using the developed method, the reactions catalyzed by the enzymes belonging to six different EC classes were analyzed, and Michaelis constants for their substrates were determibed. Km values obtained using the proposed method were in good agreement with those obtained with standard colorimetric and fluorimentric assays. Practical potential of the described method was demonstrated by studying the interactions of a diagnostically significant enzyme α-D-galactosidase with its natural and artificial substrates and its inhibitor. Km values for α-D-galactosidase using melibiose and raffinose as substrates and IC50 value for the enzyme inhibitor 1-deoxygalactonojirimycin were determined. The described method allows rapid and label-free investigation of enzyme interactions with substrates, inhibitors and activators for a wide range of biocatalysts.

The role of substrata width and millimeter scale surface micro-structure in the attachment of juvenile mussels

The global mussel aquaculture industry is constrained by the unreliable supply of wild seed mussels and high losses of seed mussels shortly after they are seeded onto coastal farms. The success in the collection of the settling larvae of mussel in the wild and the subsequent on-growing of early seed mussels in aquaculture are largely determined by the settlement and attachment behaviour of the larval and juvenile mussels, which is strongly influenced by the physical structure of their attachment substrata, especially filamentous substrata. This study aimed to identify an ideal set of morphological characteristics of filamentous substrata that would promote the improved attachment of early mussels and contribute to reducing the shortage of seed mussels in the New Zealand green-lipped mussel (Perna canaliculus) aquaculture industry. Artificial filamentous substrata with varying branch width and textured surfaces were tested for their ability to promote the attachment of wild juvenile mussels. The attachment of early juvenile mussels to 1.6 mm wide filamentous substrate was six times higher than their wider counterparts (i.e., 3.7, 5.6, 7.4, and 9.5 mm). There was also a higher proportion of small-size mussels (<0.99 mm shell length) on the 1.6 mm substrata. Furthermore, the filamentous substrata with regularly ridged surface contours (1 mm spacings) tended to attract more juvenile mussels. However, this preference was not consistent for small-size mussels on filaments of thinner width (i.e., 0.4, 0.8, 1.6, and 3.7 mm). Overall, the dimensions of filamentous substrates identified through this study will be valuable for improving the design of artificial substrata used in mussel aquaculture for improving its effectiveness for the attachment of seed mussels.

Rational Engineering of a β-Glucosidase (H0HC94) from Glycosyl Hydrolase Family I (GH1) to Improve Catalytic Performance on Cellobiose.

The conversion of lignocellulosic feedstocks by cellulases to glucose is a critical step in biofuel production. β-Glucosidases catalyze the final step in cellulose breakdown, producing glucose, and are often the rate-limiting step in biomass hydrolysis. The specific activity of most natural and engineered β-glucosidase is higher on the artificial substrate p-nitrophenyl β-d-glucopyranoside (pNPGlc) than on the natural substrate, cellobiose. We report an engineered β-glucosidase (Q319A H0HC94) with a 1.8-fold higher specific activity (366.3 ± 36 μmol/min/mg), a 1.5-fold increase in kcat (340.8 ± 27 s-1), and a 3-fold increase in catalytic efficiency (236.65 mM-1 s-1) over H0HC94 (WT) on cellobiose. Molecular dynamic simulations and protein structure network analysis indicate that the Q319A H0HC94 active site pocket is significantly remodeled compared to the WT, leading to changes in enzyme conformation, better accessibility of cellobiose inside the active site pocket, and higher enzymatic activity. This study shows the impact of rational engineering of a nonconserved residue to increase β-glucosidase substrate accessibility and catalytic efficiency by reducing crowding interaction between cellobiose and active site pocket residues near the gatekeeper region and increasing pocket volume and surface area. Thus, rational engineering of previously characterized enzymes could be an excellent strategy to improve cellulose hydrolysis.

Comparison of periphyton growth on two artificial substrates in temperate zone fishponds

Periphyton communities are fundamental components of freshwater ecosystems that influence water quality, nutrient cycling, and productivity. Additionally, periphyton communities serve as valuable food sources in aquaculture. The periphyton communities on two substrates—heather mats and geotextiles—were compared in four temperate fishponds. After 51 days of immersion, the geotextiles exhibited significantly greater fluorescence, indicating a greater abundance of algae and cyanobacteria. Conversely, the abundance (and biomass) of periphyton invertebrates on heather mats was greater than that on geotextiles, and the difference became more pronounced over time. These differences resulted from significantly greater dipteran larval abundances (biomasses) on the heather mats than on the geotextiles. This study suggested that heather mats can significantly enhance food abundance and availability within the water column under Central European conditions.

Contribution of groundwater-borne nutrients to eutrophication potential and the share of benthic algae in a large lowland river

Groundwater inflow can be a significant source of nutrients for riverine ecosystems, which can affect eutrophication i.e., the elevated primary production and the corresponding accumulation of algal biomass. Experimental and modelling work has shown that benthic algae (autotrophic biofilms) in particular benefit, as they have direct access to the inflowing groundwater-borne nutrients. Primarily the supply of phosphorus (P) enhances pelagic algal biomass, as it is the limiting nutrient for primary production in most freshwater systems. In this study, we estimate the effect of groundwater inflow on overall eutrophication of a large, European lowland river and tested its seasonal effect on biofilms in particular. We calculated the effects on overall eutrophication during summer according to the estimated input of groundwater-borne P and the C:P stoichiometry of planktonic algae in the Elbe River. Our model indicated that these diffuse P inputs have the potential to significantly increase eutrophication. Groundwater-P can contribute up to 1.5 t/d PO4 over the investigated 450 km stretch of the Elbe River under low flow conditions. This would result in an additional planktonic load of about 46 t/d of particulate organic carbon, thereby contributing to eutrophication at the regional scale in this river. In contrast, at the local scale, biofilms were collected seasonally from artificial substrata exposed in the river either in hydrogeologically active areas with groundwater inflow, or in areas of varying hydraulic connectivity. Analyses of biofilm macronutrients, structural components and biofilm community composition show distinct effects of season, hydrogeology and groundwater inflow. The dominant predictors were season and the interaction between hydrogeology and groundwater. Benthic eutrophication is most likely to occur in autumn in areas of loose rock with high groundwater inflow. The strong interaction of environmental factors in determining benthic eutrophication highlights the need to assess these factors in combination rather than in isolation.

Habitat-specific response of fish assemblages in a small fully protected urban MPA

Abstract This study aims at assessing the reserve effect on fish assemblage in a small urban fully protected and highly enforced Marine Protected Area (MPA), Larvotto MPA (Monaco, Mediterranean Sea). The data about fish density, size, and biomass was collected by underwater visual census in the MPA and unprotected areas in two habitats, artificial rocky substrates and Posidonia oceanica meadows. On artificial rocky substrates, we recorded significantly higher fish biomass within the MPA compared to unprotected areas, while no significant difference was detected on Posidonia, with this suggesting the habitat-specific responses of fish assemblages to protection. Here we highlight the potential effectiveness of highly enforced small-sized urban MPAs, such as the Larvotto MPA, in generating ecological benefits, and speculate about their potential role in supporting networks of MPAs. This study supports increased attention to urban MPAs in conservation efforts, emphasizing the need for efficient management strategies in the face of ongoing coastal expansion.

Orthogonal Persulfide Generation through Precision Tools Provides Insights into Mitochondrial Sulfane Sulfur.

The sulfane sulfur pool, comprised of persulfide (RS-SH) and polysulfide (RS-SnH) derived from hydrogen sulfide (H2S), has emerged as a major player in redox biochemistry. Mitochondria, besides energy generation, serve as significant cellular redox hubs, mediate stress response and cellular health. However, the effects of endogenous mitochondrial sulfane sulfur (MSS) remain largely uncharacterized as compared with their cytosolic counterparts, cytosolic sulfane sulfur (CSS). To investigate this, we designed a novel artificial substrate for mitochondrial 3-mercaptopyruvate sulfurtransferase (3-MST), a key enzyme involved in MSS biosynthesis. Using cells expressing a mitochondrion-localized persulfide biosensor, we demonstrate this tool's ability to selectively enhance MSS. While H2S was previously known to suppress human immunodeficiency virus (HIV-1), we found that MSS profoundly affected the HIV-1 life cycle, mediating viral reactivation from latency. Additionally, we provide evidence for the role of the host's mitochondrial redox state, membrane potential, apoptosis, and respiration rates in managing HIV-1 latency and reactivation. Together, dynamic fluctuations in the MSS pool have a significant and possibly conflicting effect on HIV-1 viral latency. The precision tools developed herein allow for orthogonal generation of persulfide within both mitochondria and the cytosol and will be useful in interrogating disease biology.

Comprehensive DNA metabarcoding-based detection of non-indigenous invertebrates in recreational marinas through a multi-substrate approach

eDNA metabarcoding has been increasingly employed in the monitoring of marine invertebrate non-indigenous species (NIS), in particular using filtered seawater. However, comprehensive detection of all NIS may require a diversity of sampling substrates. To assess the effectiveness of 5 sample types (hard and artificial substrates, water, zooplankton) on the recovery of invertebrates’ diversity, two marinas were monitored over three time points, using COI and 18S rRNA genes as DNA metabarcoding markers. We detected a total of 628 species and 23 NIS, with only up to 9% species and 17% of NIS detected by all sample types. Hard and artificial substrates were similar to each other but displayed the most significant difference in invertebrate recovery when compared to water eDNA and zooplankton. Five NIS are potential first records for Portugal. No NIS were detected in all sample types and seasons, highlighting the need for varied sampling approaches, and consideration of temporal variation for comprehensive marine NIS surveillance.

Comparison of in situ black-lipped oyster spat collection and larval dispersal modelling results in semi-closed pearl-farming lagoons of the Tuamotu Archipelago

Spat collection of the pearl oyster Pinctada margaritifera in atoll lagoons of French Polynesia is the fundamental sustain of black pearl farming. Spat collection has always yielded variable results in space and time, but obvious signs of steady decreases, even collapses, have emerged in several lagoons. Spat collection materializes the ecological connectivity pathways between wild spawning populations and the location of artificial larval settlement substrates. To assess if oyster larval dispersal modelling could capture such pathways, we compared four six-week long spat collector deployment periods with dispersal simulations in two different lagoons. Spat collectors displayed wide spatial and temporal variations. Numerical modelling and field experiments were generally not in agreement. Although both methods have limitations, they can still approximate each other. But the accuracy of model simulations cannot be ascertained with spat collection data only. Using a SWOT (Strength-Weakness-Opportunities-Threats) analysis, we emphasize the complementarity of both approaches for management decisions.

Musseling through: Mytilus byssal thread production is unaffected by continuous noise

Anthropogenic low-frequency noise (ALFN) is a rising pollutant in the world oceans. Despite the ubiquity of ALFN, its effect on marine invertebrates is still poorly understood. Here, we tested how continuous low-frequency noise (CLFN), a substantial component of ALFN, affects the byssal thread production of Mytilus, a cosmopolitan genus of mussels with high ecological and economic importance. The effects of acute CLFN exposure and predator cues on byssogenesis by Mytilus spp. were explored in both the presence and absence of predator cues. While predator effluents increased thread production, CLFN had seemingly no effect on thread counts. Further, trends suggested a synergistic effect of CLFN and predator cues. The behavioral indifference of Mytilus spp. toward CLFN could contribute to the observed prevalence of these animals in inherently disturbed habitats. This would partly explain their success in colonizing and persisting on artificial substrata rife with disturbances.

Factors influencing the epibiota of granite blocks used in coastal defense structures: An observational experiment

Anthropogenic pressures exerted on coastal areas, combined with climate change, and accelerated sea level rise, result in the loss of natural habitats. At the same time, the number of coastal protection and defense structures have increased considerably and may become the dominant habitat in many intertidal regions. In this study we characterize the epibiota of artificial substrates (granite blocks) from coastal defense structures used to contain the effects of rising sea levels, and the same type of artificial substrate found in contact with coastal reefs located on Serrambi beach, northeast of Brazil. We mapped, measured, and photographed 45 artificial granite blocks and their surroundings in the field. A gradient was found for the number of taxa on the blocks, from high coverage by macroalgae to extensive non-colonized areas. However, it was observed that the closer to the natural reef the greater the taxonomic richness. Our results also indicate that the factor that best explains the colonization of blocks is the coverage of their surroundings, thus, few organisms were found on blocks whose surrounding unconsolidated sediment area was very extensive. In our wind and sun exposure simulation, granite rock demonstrated a more accelerated and greater level of heating (up to 3 °C) compared to fragments of natural substrate, although the variation in the slightly lighter colored and smaller granite block resembles the variation in the reef fragment. We suggest that the use of lighter-colored granite blocks at mid-coastal levels can increase the colonization rate of these substrates.

The Bifidobacterium adolescentis BAD_1527 gene encodes GH43_22 α-L-arabinofuranosidase of AXH-m type

Bifidobacterium adolescentis gene BAD_1527 has previously been suggested to code for a β-xylosidase (Kobayashi et al., Mar Drugs 18:174, 2020). Our detailed investigation of the substrate specificity of the GH43_22 protein using a wide spectrum of natural and artificial substrates showed that the enzyme hydrolyzed neither linear xylooligosaccharides nor glucuronoxylan. Xylose was released only from the artificial 4-nitrophenyl β-D-xylopyranoside (1.58 mU/mg). The corresponding α-L-arabinofuranoside was by three orders of magnitude better substrate (2.17 U/mg). Arabinose was the only monosaccharide liberated from arabinoxylan and α-1,3- or α-1,2-singly arabinosylated xylooligosaccharides. Moreover, the enzyme efficiently debranched sugar beet arabinan and singly arabinosylated α-1,5-L-arabinooligosaccharides, although short linear α-1,5-L-arabinooligosaccharides were also slowly degraded. On the other hand, debranched arabinan, arabinogalactan as well as 2,3-doubly arabinosylated main chain residues of arabinan and arabinoxylan did not serve as substrates. Thus, the enzyme encoded by the BAD_1527 gene is a typical α-L-arabinofuranosidase of AXH-m specificity.

Identification of Orthosteric and Allosteric Pharmacological Chaperones for Mucopolysaccharidosis Type IIIB.

Mucopolysaccharidosis type IIIB (MPS IIIB) is an autosomal inherited disease caused by mutations in gene encoding the lysosomal enzyme N-acetyl-alpha-glucosaminidase (NAGLU). These mutations result in reduced NAGLU activity, preventing it from catalyzing the hydrolysis of the glycosaminoglycan heparan sulfate (HS). There are currently no approved treatments for MPS IIIB. A novel approach in the treatment of lysosomal storage diseases is the use of pharmacological chaperones (PC). In this study, we used a drug repurposing approach to identify and characterize novel potential PCs for NAGLU enzyme. We modeled the interaction of natural and artificial substrates within the active cavity of NAGLU (orthosteric site) and predicted potential allosteric sites. We performed a virtual screening for both the orthosteric and the predicted allosteric site against a curated database of human tested molecules. Considering the binding affinity and predicted blood-brain barrier permeability and gastrointestinal absorption, we selected atovaquone and piperaquine as orthosteric and allosteric PCs. The PCs were evaluated by their capacity to bind NAGLU and the ability to restore the enzymatic activity in human MPS IIIB fibroblasts These results represent novel PCs described for MPS IIIB and demonstrate the potential to develop novel therapeutic alternatives for this and other protein deficiency diseases.

ROPE TO THE SEA: OBSERVATION OF THE FISHING ROPE ACTING AS ARTIFICIAL SUBSTRATA TO FIXATION OF CORAL Astrangia solitaria (LE SUEUR, 1817) COLLECTED IN THE GREAT AMAZON REEF SYSTEM (GARS)

Herein, we report the first observation of the coral Astrangia solitaria (Le Sueur, 1817) adhered on the polypropylene fishing rope (artificial substrate), collected in the areas of the Great Amazon Reef System (GARS). The specimens of A. solitaria were collected adhered on the fishing rope abandoned in bottom areas associated with the GARS, in the state of Amapá (Northern Brazil) (02°57'54"N; 048°27'50,4"W), during the commercial fishing operations of the red snapper - Lutjanus purpureus (Poey, 1866) - in May 2024. We observed 226 m of lost rope from the illegal lobster trap “caçoeira” adhered (wrapped) in the fish trap called “manzuá”, where we accounted 63 colonies of A. solitaria covering 12.5 cm of this rope. This observation can be associated with a wide range of marine invertebrate adaptations for the use of these types of macroplastics in an anthropized environment.

Unveiling the role of taxonomic sufficiency for enhanced ecosystem monitoring

The use of Artificial substrates (AS) as sampling devices addresses challenges in macrofaunal quantitative sampling. While effectively capturing biodiversity patterns, the time-intensitive identification process at the species level remains a substantial challenge. The Taxonomic Sufficiency approach (TS), where only taxa above species level are identified, arises as a potential solution to be tested across different environmental monitoring scenarios. In this paper, we analyzed three AS macrobenthic datasets to evaluate the odds of TS in improving the cost-effective ratio in AS monitoring studies and establish the highest resolution level to detect assemblage changes under different environmental factors. Results indicated that the family level emerged as a pragmatic compromise, balancing precision and taxonomic effort. Cost/benefit analysis supported TS efficiency, maintaining correlation stability until the family level. Results also showed that reducing resolution to family does not entail a significant Loss of Information. This study contributes to the discourse on TS applicability, highlighting its practicality in monitoring scenarios, including spatial-temporal studies, and rapid biodiversity assessments. Additionally, it highlights the “second best approach” of family-level practicality depending on the specific monitoring scenario and recognizes the importance of the species-level “best approach” before applying TS in monitoring studies.

Physical structure of the environment contributes to the development of diversity of microalgal assemblages

Aquatic macrophytes form a three dimensional complex structure in the littoral zones of lakes, with many physical, chemical and biological gradients and interactions. This special habitat harbours a unique microalgal assemblage called metaphyton, that differs both from the phytoplankton of the pelagial and from the benthic assemblages whose elements are tightly attached to the substrates. Since metaphytic assemblages significantly contribute to the diversity of lakes’ phytoplankton, it is crucial to understand and disentangle those mechanisms that ensure their development. Therefore, we focused on the question of how a single solid physical structure contribute to maintaining metaphytic assemblages. Using a laboratory experiment we studied the floristic and functional differences of microalgal assemblages in microcosms that simulated the conditions that an open water, a complex natural macrophyte stand (Utricularia vulgaris L.), or an artificial substrate (cotton wool) provide for them. We inoculated the systems with a species rich (> 326 species) microalgal assemblage collected from a eutrophic oxbow lake, and studied the diversity, trait and functional group composition of the assemblages in a 24 day long experimental period. We found that both natural and artificial substrates ensured higher species richness than the open water environment. Functional richness in the open water environment was lower than in the aquaria containing natural macrophyte stand but higher than in which cotton wool was placed. This means that the artificial physical structure enhanced functional redundancy of the resident functional groups. Elongation measures of microalgal assemblages showed the highest variation in the microcosms that simulated the open water environment. Our results suggest that assembly of metaphytic algal communities is not a random process, instead a deterministic one driven by the niche characteristics of the complex three dimensional structure created by the stands of aquatic macrophytes.