The Response of Extracellular Polymeric Substances Production by Phototrophic Biofilms to a Sequential Disturbance Strongly Depends on Environmental Conditions

Phototrophic biofilms are exposed to multiple stressors that can affect them both directly and indirectly. By modifying either the composition of the community or the physiology of the microorganisms, press stressors may indirectly impact the ability of the biofilms to cope with disturbances. Extracellular polymeric substances (EPS) produced by the biofilm are known to play an important role in its resilience to various stresses. The aim of this study was to decipher to what extent slight modifications of environmental conditions could alter the resilience of phototrophic biofilm EPS to a realistic sequential disturbance (4-day copper exposure followed by a 14-day dry period). By using very simplified biofilms with a single algal strain, we focused solely on physiological effects. The biofilms, composed by the non-axenic strains of a green alga (Uronema confervicolum) or a diatom (Nitzschia palea) were grown in artificial channels in six different conditions of light intensity, temperature and phosphorous concentration. EPS quantity (total organic carbon) and quality (ratio protein/polysaccharide, PN/PS) were measured before and at the end of the disturbance, and after a 14-day rewetting period. The diatom biofilm accumulated more biomass at the highest temperature, with lower EPS content and lower PN/PS ratio while green alga biofilm accumulated more biomass at the highest light condition with lower EPS content and lower PN/PS ratio. Temperature, light intensity, and P concentration significantly modified the resistance and/or recovery of EPS quality and quantity, differently for the two biofilms. An increase in light intensity, which had effect neither on the diatom biofilm growth nor on EPS production before disturbance, increased the resistance of EPS quantity and the resilience of EPS quality. These results emphasize the importance of considering the modulation of community resilience ability by environmental conditions, which remains scarce in the literature.

Interaction between CO2-consuming autotrophy and CO2-producing heterotrophy in non-axenic phototrophic biofilms

As the effects of climate change become increasingly evident, the need for effective CO2 management is clear. Microalgae are well-suited for CO2 sequestration, given their ability to rapidly uptake and fix CO2. They also readily assimilate inorganic nutrients and produce a biomass with inherent commercial value, leading to a paradigm in which CO2-sequestration, enhanced wastewater treatment, and biomass generation could be effectively combined. Natural non-axenic phototrophic cultures comprising both autotrophic and heterotrophic fractions are particularly attractive in this endeavour, given their increased robustness and innate O2-CO2 exchange. In this study, the interplay between CO2-consuming autotrophy and CO2-producing heterotrophy in a non-axenic phototrophic biofilm was examined. When the biofilm was cultivated under autotrophic conditions (i.e. no organic carbon), it grew autotrophically and exhibited CO2 uptake. After amending its growth medium with organic carbon (0.25 g/L glucose and 0.28 g/L sodium acetate), the biofilm rapidly toggled from net-autotrophic to net-heterotrophic growth, reaching a CO2 production rate of 60 μmol/h after 31 hours. When the organic carbon sources were provided at a lower concentration (0.125 g/L glucose and 0.14 g/L sodium acetate), the biofilm exhibited distinct, longitudinally discrete regions of heterotrophic and autotrophic metabolism in the proximal and distal halves of the biofilm respectively, within 4 hours of carbon amendment. Interestingly, this upstream and downstream partitioning of heterotrophic and autotrophic metabolism appeared to be reversible, as the position of these regions began to flip once the direction of medium flow (and hence nutrient availability) was reversed. The insight generated here can inform new and important research questions and contribute to efforts aimed at scaling and industrializing algal growth systems, where the ability to understand, predict, and optimize biofilm growth and activity is critical.

Phototrophic biofilms and microbial mats from the marine littoral of the central Mediterranean

Phototrophic biofilm and microbial mat communities grow along the rocky coastline of the Maltese islands. During this study, phototrophs from the mediolittoral and supralittoral zones were studied over a two-year period and seasonal changes were observed. Attachment of pioneer microorganisms to the porous eroded limestone bedrock was facilitated via a gelatinous matrix composed of exopolymeric substances (EPS). In submerged areas, such as undisturbed rock pools, these progressively formed green or brown compact biofilms, some of which thickened over spring to form microbial mats via the production of more extensive EPS layers. Microbial mats gradually attained a lighter colouration due to the presence of ultraviolet (UV) screening pigments. In full summer, they were observed to shrink, detach from the exposed substrate, harden and progressively calcify. Biofilm microorganisms survived the harsh summer months in sheltered areas. The major biofilm formers were filamentous non-heterocytous cyanobacteria belonging to the Leptolyngbyaceae, Pseudanabaenaceae and Oscillatoriaceae. Their sheaths were thick, lamellated and often confluent. A higher biodiversity of phototrophs was observed in late autumn and winter, when tufts of heterocytous Calothrix sp. grew on thin compact biofilms of Nodosilinea sp., Toxifilum sp. and Phormidesmis spp., while Lyngbya spp. trichomes were surrounded by thick brown sheaths. Germlings of green and brown macroalgal species belonging to Ulva, Cladophora and Sphacelaria were embedded in biofilms and microbial mats and gradually grew to form extensive macroalgal covers submerged in rock pools. Erythrotrichia sp. filaments colonised the mediolittoral zone and were confined to areas that were exposed to wave action and submerged intermittently. Over summer, macroalgal coverage diminished and microalgal biofilms and microbial mats prevailed in rock pools.

Nitrogen Source Governs Community Carbon Metabolism in a Model Hypersaline Benthic Phototrophic Biofilm

ABSTRACT Increasing anthropogenic inputs of fixed nitrogen are leading to greater eutrophication of aquatic environments, but it is unclear how this impacts the flux and fate of carbon in lacustrine and riverine systems. Here, we present evidence that the form of nitrogen governs the partitioning of carbon among members in a genome-sequenced, model phototrophic biofilm of 20 members. Consumption of NO3− as the sole nitrogen source unexpectedly resulted in more rapid transfer of carbon to heterotrophs than when NH4+ was also provided, suggesting alterations in the form of carbon exchanged. The form of nitrogen dramatically impacted net community nitrogen, but not carbon, uptake rates. Furthermore, this alteration in nitrogen form caused very large but focused alterations to community structure, strongly impacting the abundance of only two species within the biofilm and modestly impacting a third member species. Our data suggest that nitrogen metabolism may coordinate coupled carbon-nitrogen biogeochemical cycling in benthic biofilms and, potentially, in phototroph-heterotroph consortia more broadly. It further indicates that the form of nitrogen inputs may significantly impact the contribution of these communities to carbon partitioning across the terrestrial-aquatic interface. IMPORTANCE Anthropogenic inputs of nitrogen into aquatic ecosystems, and especially those of agricultural origin, involve a mix of chemical species. Although it is well-known in general that nitrogen eutrophication markedly influences the metabolism of aquatic phototrophic communities, relatively little is known regarding whether the specific chemical form of nitrogen inputs matter. Our data suggest that the nitrogen form alters the rate of nitrogen uptake significantly, whereas corresponding alterations in carbon uptake were minor. However, differences imposed by uptake of divergent nitrogen forms may result in alterations among phototroph-heterotroph interactions that rewire community metabolism. Furthermore, our data hint that availability of other nutrients (i.e., iron) might mediate the linkage between carbon and nitrogen cycling in these communities. Taken together, our data suggest that different nitrogen forms should be examined for divergent impacts on phototrophic communities in fluvial systems and that these anthropogenic nitrogen inputs may significantly differ in their ultimate biogeochemical impacts.