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The way strong environmental gradients shape multispecific assemblages has allowed us to examine a suite of ecological and evolutionary hypotheses about structure, regulation and community responses to fluctuating environments. But whether the highly diverse co-occurring microorganisms are shaped in similar ways as macroscopic organisms across the same gradients has yet to be addressed in most ecosystems. Here, we characterize intertidal biofilm bacteria communities, comparing zonation at both the "species" and community levels, as well as network attributes, with co-occurring macroalgae and invertebrates in the same rocky shore system. The results revealed that the desiccation gradient has a more significant impact on smaller communities, while both desiccation and submersion gradients (surge) affect the larger, macroscopic communities. At the community level, we also confirmed the existence of distinct communities within each intertidal zone for microorganisms, similar to what has been previously described for macroorganisms. But our results indicated that dominant microbial organisms along the same environmental gradient exhibited less differentiation across tidal levels than their macroscopic counterparts. However, despite the substantial differences in richness, size and attributes of co-occurrence networks, both macro- and micro-communities respond to stress gradients, leading to the formation of similar zonation patterns in the intertidal rocky shore. [ABSTRACT FROM AUTHOR]

Substrates like sand or gravels and aquatic nutrient concentrations of rivers are highly heterogeneous, influencing the abundance of functional genes in epilithic biofilms where nitrification–denitrification processes take place. To analyze how the relative abundance of nitrifying/denitrifying genes and the associated microbes changes with the physical properties of substrates and aquatic concentrations of nutrients, this paper utilized metagenomics to comprehensively characterize these functional genes (i.e., amoA, hao, and nxrB involved in nitrification, and napA, narG, nirS, norB, and nosZ associated with denitrification) from epilithic biofilms collected along the Shitingjiang River in Southwest China and further obtained the relative abundance of major nitrifiers and denitrifiers. The results show that substrate size most significantly affects the relative abundance of hao and norB by altering the hydrodynamic conditions. In sampling sites with high heterogeneity in substrate size distribution, the relative abundance of most denitrifying genes is also higher. The carbon–nitrogen ratio negatively correlates with the relative abundance of all the nitrifying genes, while ammonium, total inorganic carbon, and total organic carbon concentrations positively affect the relative abundance of amoA and nxrB. As to the relative abundance of nitrifiers and denitrifiers, mainly belonging to phyla Proteobacteria and Actinobacteria, substrate heterogeneity and the aquatic concentrations of nutrients have greater influences than substrate size. Also, the substrate heterogeneity exerted positive influence on functional species of Pseudogemmobacter bohemicus and Paracoccus zhejiangensis. Considering the genes' functions and the dominant species linked to denitrification, nitrous oxide is more likely to occur in rivers with higher heterogeneity and larger substrates. [ABSTRACT FROM AUTHOR]

The ability of WWTP outflow bacteria at colonizing rock surfaces and contributing to the formation of river epilithic biofilms was investigated. Bacterial community structures of biofilms (b-) developing on rocks exposed to treated wastewaters (TWW) of a hospital (HTWW) and a domestic (DTWW) clarifier, and to surface waters of the stream located at 10 m, 500 m, and 8 km from the WWTP outlet, were compared. Biofilm bacterial contents were analyzed by cultural approaches and a tpm-based DNA metabarcoding analytical scheme. Co-occurrence distribution pattern analyses between bacterial datasets and eighteen monitored pharmaceuticals were performed. Higher concentrations of iohexol, ranitidine, levofloxacin, and roxithromycin were observed in the b-HTWW while atenolol, diclofenac, propranolol, and trimethoprim were higher in the b-DTWW. MPN growth assays showed recurrent occurrences of Pseudomonas aeruginosa and Aeromonas caviae among these biofilms. An enrichment of multi-resistant P. aeruginosa cells was observed in the hospital sewer line. P. aeruginosa MPN values were negatively correlated to roxithromycin concentrations. The tpm DNA metabarcoding analyses confirmed these trends and allowed an additional tracking of more than 90 species from 24 genera. Among the recorded 3082 tpm ASV (amplicon sequence variants), 41% were allocated to the Pseudomonas. Significant differences through ANOSIM and DESeq2 statistical tests were observed between ASV recovered from b-HTWW, b-DTWW, and epilithic river biofilms. More than 500 ASV were found restricted to a single sewer line such as those allocated to Aeromonas popoffii and Stenotrophomonas humi being strictly found in the b-HTWW file. Several significant correlations between tpm ASV counts per species and pharmaceutical concentrations in biofilms were recorded such as those of Lamprocystis purpurea being positively correlated with trimethoprim concentrations. A tpm source tracking analysis showed the b-DTWW and b-HTWW tpm ASV to have contributed, respectively, at up to 35% and 2.5% of the epilithic river biofilm tpm-taxa recovered downstream from the WWTP outlet. Higher contributions of TWW taxa among epilithic biofilms were recorded closer to the WWTP outlet. These analyses demonstrated a coalescence of WWTP sewer communities with river freshwater taxa among epilithic biofilms developing downstream of a WWTP outlet. [ABSTRACT FROM AUTHOR]

In forested headwater streams, inconspicuous food resources such as epilithic microalgae can play a major role owing to their content of long chain polyunsaturated fatty acids (PUFAs) that are essential for macroinvertebrate development. Yet, the use of these resources and their consequences for consumers and life‐history traits remain scarcely studied, especially for non‐herbivorous taxa.Using instream mesocosms, we aimed to understand how two macroinvertebrate species, a shredder detritivore (Gammarus pulex) and a scraper (Rhithrogena semicolorata), use available organic resources under light versus shaded conditions (high and low amounts of phototrophic biofilm, respectively). We specifically focused on the origin of carbon (C) and essential lipids (PUFAs, sterols) assimilated by the two species and the effects of these compounds on their survival and growth.When autotrophic biofilms were available (mesocosms exposed to light), both species experienced significantly higher growth than in mesocosms placed in the dark. In Rhithrogena nymphs, the survival and imago emergence rates were positively affected by access to autotrophic biofilms.Using stable isotope analysis (δ13C and δ15N), we demonstrated that under dark conditions both macroinvertebrates assimilated C of detrital origin. Under light conditions, most of the C assimilated by R. semicolorata was derived from autotrophic biofilms and around 11% from fine detrital particles (FPOM). Gammarus pulex derived part of its C from detrital sources (mainly FPOM) and from autotrophic biofilms.The results of isotopic analyses (δ13C) on fatty acids (FAs) and sterols showed that the scraper Rhithrogena was entirely dependent on autotrophic biofilms to meet its dietary FAs and sterol requirements. In contrast, detrital sources were quantitatively important for G. pulex both in terms of C and sterol supply, irrespective of the conditions tested. For these consumers, microalgae seemed to be a complementary food source, but yet essential to cover a large part of their requirements in long‐chain PUFAs.This study clearly confirmed the ecological importance of autotrophic biofilms for two functional feeding groups of macroinvertebrates in headwater streams providing new insights on the trophic origin of sterols and long‐chain PUFAs in their diet. [ABSTRACT FROM AUTHOR]

The ability of WWTP outflow bacteria at colonizing rock surfaces and contributing to the formation of river epilithic biofilms was investigated. Bacterial community structures of biofilms (b-) developing on rocks exposed to treated wastewaters (TWW) of a hospital (HTWW) and a domestic (DTWW) clarifier, and to surface waters of the stream located at 10 m, 500 m, and 8 km from the WWTP outlet, were compared. Biofilm bacterial contents were analyzed by cultural approaches and a tpm-based DNA metabarcoding analytical scheme. Co-occurrence distribution pattern analyses between bacterial datasets and eighteen monitored pharmaceuticals were performed. Higher concentrations of iohexol, ranitidine, levofloxacin, and roxithromycin were observed in the b-HTWW while atenolol, diclofenac, propranolol, and trimethoprim were higher in the b-DTWW. MPN growth assays showed recurrent occurrences of Pseudomonas aeruginosa and Aeromonas caviae among these biofilms. An enrichment of multi-resistant P. aeruginosa cells was observed in the hospital sewer line. P. aeruginosa MPN values were negatively correlated to roxithromycin concentrations. The tpm DNA metabarcoding analyses confirmed these trends and allowed an additional tracking of more than 90 species from 24 genera. Among the recorded 3082 tpm ASV (amplicon sequence variants), 41% were allocated to the Pseudomonas. Significant differences through ANOSIM and DESeq2 statistical tests were observed between ASV recovered from b-HTWW, b-DTWW, and epilithic river biofilms. More than 500 ASV were found restricted to a single sewer line such as those allocated to Aeromonas popoffii and Stenotrophomonas humi being strictly found in the b-HTWW file. Several significant correlations between tpm ASV counts per species and pharmaceutical concentrations in biofilms were recorded such as those of Lamprocystis purpurea being positively correlated with trimethoprim concentrations. A tpm source tracking analysis showed the b-DTWW and b-HTWW tpm ASV to have contributed, respectively, at up to 35% and 2.5% of the epilithic river biofilm tpm-taxa recovered downstream from the WWTP outlet. Higher contributions of TWW taxa among epilithic biofilms were recorded closer to the WWTP outlet. These analyses demonstrated a coalescence of WWTP sewer communities with river freshwater taxa among epilithic biofilms developing downstream of a WWTP outlet.

Nitrogen (N) uptake is a key process in stream ecosystems that is mediated mainly by benthic microorganisms (biofilms on different substrata) and has implications for the biogeochemical fluxes at catchment scale and beyond. Here, we focused on the drivers of assimilatory N uptake, especially the effects of hydromorphology and other environmental constraints, across three spatial scales: micro, meso and reach. In two seasons (summer and spring), we performed whole-reach 15N-labelled ammonium injection experiments in two montane, gravel-bed stream reaches with riffle–pool sequences. N uptake was highest in epilithic biofilms, thallophytes and roots (min–max range 0.2–545.2 mg N m−2 day−1) and lowest in leaves, wood and fine benthic organic matter (0.05–209.2 mg N m−2 day−1). At the microscale, N uptake of all primary uptake compartments except wood was higher in riffles than in pools. At the mesoscale, hydromorphology determined the distribution of primary uptake compartments, with fast-flowing riffles being dominated by biologically more active compartments and pools being dominated by biologically less active compartments. Despite a lower biomass of primary uptake compartments, mesoscale N uptake was 1.7–3.0 times higher in riffles than in pools. At reach scale, N uptake ranged from 79.6 to 334.1 mg N m−2 day−1. Highest reach-scale N uptake was caused by a bloom of thallopyhtes, mainly filamentous autotrophs, during stable low discharge and high light conditions. Our results reveal the important role of hydromorphologic sorting of primary uptake compartments at mesoscale as a controlling factor for reach-scale N uptake in streams. [ABSTRACT FROM AUTHOR]

Active functional groups in biofilms determine the adsorption and desorption of contaminants and nutrients. Epilithic biofilms were characterized in order to understand the association between the chemistry alteration patterns and the surrounding anthropic activities of the Guaporé River watershed. The instrumental analyses included pyrolysis coupled to gas chromatography and mass spectroscopy, spectroscopy in the IR region with attenuated total reflectance, and two-dimensional nuclear magnetic resonance. Spectrometric techniques demonstrated that epilithic biofilms are mainly composed of polysaccharides, nitrogen-containing compounds, lipids, and aromatic and phenolic structures, which have functional groups characteristic of alcohols, esters, ethers, and amides. The polysaccharide levels reflect well the environmental pressures. The chemical composition of epilithic biofilms can be an effective tool for environmental assessment in watercourses, since the different anthropic actions developed in watersheds, mainly agriculture and urban areas, can modify the organic fraction of biofilms. [ABSTRACT FROM AUTHOR]

Denitrification is the major process of nitrate reduction to dinitrogen (N2). This is the first report on detection of the nirK type denitrifying bacteria in the epilithic biofilms of the Lake Baikal littoral zone. The highest species diversity was revealed in microbial communities of the biofilms collected in the area of the Bol'shoe Goloustnoe settlement and Severobaikal'sk City, while those collected in the vicinity of Listvyanka settlement exhibited the lowest diversity. Phylogenetic analysis showed uncultured denitrifying bacteria from diverse habitats were most closely related to all cloned sequences. The closely related homologs were represented by the sequences of the orders Rhizobiales and Rhodobacterales, class Alphaproteobacteria. [ABSTRACT FROM AUTHOR]

Benthic biofilms are often assumed to control terrestrially-derived dissolved organic carbon (tDOC) uptake in streams. We tested this by comparing 13C-enriched ryegrass leachate uptake in an agricultural and a forest stream, hypothesizing that a greater abundance of autotrophic biofilms in the agricultural stream would cause its whole-stream tDOC uptake to be comparatively low. We measured whole-stream and biofilm tDOC tracer uptake, metabolism, bacterial and algal diversity, and nutrient status of benthic epilithic biofilms, and assessed whole-stream hydromorphology. Whole-stream uptake of tDOC was six times lower in the agricultural (3.0 mg m−2 day−1) than in the forest (19.0 mg m−2 day−1) stream, and tDOC uptake velocity indicated lower tDOC demand in the agricultural (1.2 mm min−1) than in the forest (1.9 mm min−1) stream. The agricultural stream differed from the forest stream by slightly lower transient storage capacity and higher benthic biofilm bacterial abundance and production, lower biofilm biomass and lower biofilm molar C:N, C:P, and N:P ratios. Changes in epilithic biofilms contributed little to the differences in whole-stream tDOC tracer uptake between streams, as biofilm tDOC uptake only amounted to 4% and 13% of whole-stream uptake in the forest and agricultural stream, respectively. This comparison of a forest and an agricultural stream suggests that agricultural stressors have the potential to diminish both whole-stream tDOC uptake and uptake efficiency. Furthermore, the weak link between biofilm and whole-stream tDOC uptake implies that benthic biofilms characteristics are poor predictors for human impacts on tDOC uptake in agricultural streams and that hot spots of tDOC uptake are likely situated in the hyporheic zone or in the stream water column. [ABSTRACT FROM AUTHOR]

Abstract Epilithic biofilms are communities of microorganisms composed mainly of microbial cells, extracellular polymeric substances from the metabolism of microorganisms, and inorganic materials. Biofilms are a useful tool to assess the impact of anthropic action on aquatic environments including the presence of pesticide residues such as glyphosate. The present work seeks to monitor the occurrence of glyphosate and AMPA residues in epilithic biofilms occurring in a watershed. For this, epilithic biofilm samples were collected in the Guaporé River watershed in the fall and spring seasons of 2016 at eight points. Physicochemical properties of the water and biofilms were determined. The determination of glyphosate and AMPA was performed using an ultra-high performance liquid chromatograph coupled to a tandem mass spectrometer. The concentrations of glyphosate and AMPA detected in epilithic biofilms vary with the season (from 90 to 305 μg kg−1 for glyphosate and from 50 to 240 μg kg−1 for AMPA, in fall and spring, respectively) and are strongly influenced by the amount of herbicide applications. Protected locations and those with poor access not demonstrate the presence of these contaminants. In the other seven points of the Guaporé River watershed, glyphosate was detected in concentrations ranging from 10 to 305 μg kg−1, and concentrations of AMPA ranged from 50 to 670 μg kg−1. An overview of the contamination in the Guaporé watershed shows that the most affected areas are located in the Marau sub-watershed, which are strongly influenced by the presence of the city of Marau. This confirms the indiscriminate use of glyphosate in the urban area (weed control, domestic gardens and horticulture) and constitutes a problem for human and animal health. The results showed that biofilms can accumulate glyphosate resulting from the contamination of water courses and are sensitive to the sources of pollution and pesticides present in rivers. Graphical abstract Unlabelled Image Highlights • Epilithic biofilms are accumulators of glyphosate and AMPA. • Epilithic biofilms are able to differentiate anthropic pressures. • Glyphosate and AMPA concentrations vary with seasons. • The indiscriminate use of glyphosate constitutes a problem for human health. [ABSTRACT FROM AUTHOR]

The Ave River in northern Portugal has a history of riverbanks and water quality degradation. The river water quality was assessed by physicochemical, biological (macroinvertebrates) and microbiological (Enterococcus spp. and Escherichia coli) parameters in six locations (A-F, point A being the nearest to the source) throughout its course during a year. Epilithic biofilms were studied through polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE). Antimicrobial susceptibility testing helped with selecting isolates (n ¼ 149 E. coli and n ¼ 86 enterococci) for further genetic characterization. Pursuant to physicochemical and macroinvertebrates-based parameters, the river water was of reasonable quality according to European legislation (Directive 2000/60/EC). However, the microbiological analysis showed increased fecal contamination downstream from point C. At point D, four carbapenem-resistant E. coli isolates were recovered. Paradoxically, point D was classified as a point of 'Good Water Quality' according to macroinvertebrates results. Point F presented the highest contamination level and incidence of multidrug-resistant (MDR) isolates in the water column (13 MDR enterococci out of 39 and 33 MDR E. coli out of 97). Epilithic biofilms showed higher diversity in pristine points (A and B). Thus, biological and microbiological parameters used to assess the water quality led to divergent results; an outcome that reinforces the need for a holistic evaluation. [ABSTRACT FROM AUTHOR]

Abstract: Differences in the quantitative and qualitative compositions of various physiological and biochemical groups of cultured microorganisms were revealed in biofilms formed on stony substrates in the littoral zone of Southern Baikal (August 2012) in the areas with different anthropogenic load. Maximal abundance of microorganisms was registered in epilithic biofilms from the area of Baikalsk Pulp and Paper Plant: organotrophs (770.2 ± 290.3 × 103 CFU/cm2); amylolytics (38.1 ± 7.7 × 103 CFU/cm2); phosphate-solubilizing microorganisms (4.8 ± 1.4 × 103 CFU/cm2); phosphate-mobilizing microorganisms (212.0 ± 27.6 × 103 CFU/cm2); phenol-oxidizing microorganisms (15.7 ± 2.4 × 103 CFU/cm2); molds (3.2 ± 0.4 × 103 CFU/cm2); and total coliform bacteria (193.3 ± 28.2 CFU/cm2), with the total microbial abundance (TMA at 37°C) of 24.9 ± 1.7 × 103 CFU/cm2, which indicated high degrading activity of the biofilms in response to the anthropogenic load in the area. Epilithic biofilms of the Bol'shie Koty area exhibited the lowest abundance of organotrophic and phosphate-solubilizing microorganisms, while molds were absent. [ABSTRACT FROM AUTHOR]

Many bacterial secondary metabolites, including pharmacologically promising compounds, are synthesized by polyketide synthases (PKS) enzyme complexes. In the present work, nucleotide sequences of the genes encoding 16S rRNA and PKS of heterotrophic bacterial strains isolated from epilithic biofilms of the littoral zone of Lake Baikal were determined. On the basis of molecular phylogenetic analysis of the 16S rRNA genes, six heterotrophic strains were identified: Serratia fonticola 1А and 10А, Pseudomonas umsongensis K10-2 and K10-3, Rheinheimera tilapiae K18, and Flavobacterium sp. 43-09. Sequencing of cloned amplification products for PKS gene cluster revealed 33 sequences. Genes involved in biosynthesis of antibiotics (difficidine, erythromycin, curacin, mixalamide, corallopyronin, and myxothiazol) and cytostatics (romidepsin, spiruchostatin, and disorazol) were found among homologous sequences. The low homology (50-83%) of the PKS amino acid sequences of Baikal bacteria with sequences from GenBank attests to the potential capability of strains to produce new, not yet studied bioactive compounds. The obtained results show that the studied strains may be of practical interest for biotechnological application. [ABSTRACT FROM AUTHOR]

We studied biochemical changes in biofilm and suspended particulate and dissolved organic matter (OM) during the leaf emergence period (March-May 2008) in a forested headwater stream in response to a long-term (4 years, 2004-2008) experimental nutrient enrichment study. This study compared results from one reach upstream of the enrichment point and one reach downstream using moderate nutrient concentrations (nitrogen, N, from 388 to 765 μg L and phosphorus, P, from 10 to 30 μg L, resulting in N:P ratios of 85-56). During the spring of 2008, we analysed the chlorophyll content, elemental composition (carbon, C, and N), bacterial density, and extracellular enzyme activities along with their biochemical composition (amino acids, fatty acids and sterols) on biofilm and OM. Nutrients caused changes in the biochemical composition of the biofilm, while changes in the OM were subtle. The C:N ratio of the biofilm decreased with nutrient enrichment likely due to the increase in protein (non-essential amino acids). The polysaccharide and total and essential fatty acid contents were higher when nutrient enrichment coincided with greater light availability. The peptidase extracellular activity was higher in the fertilised reach at early spring, while phosphatase activity decreased at late spring. The suspended and dissolved OM composition did not change due to the nutrient addition, likely due to the lower water residence time in the reach. Headwater systems are highly dynamic, and the biochemical composition of the biofilm changed in response to changes in nutrients but also to light in this study. These changes, although moderate, could influence higher trophic levels through modifications in their diet. This experiment exemplifies how small land use shifts may affect headwater streams. [ABSTRACT FROM AUTHOR]

Soil and rock surfaces support microbial communities involved in mineral weathering processes. Using selective isolation, fungi were obtained from limestone surfaces of Mayan monuments in the semi-arid climate at Yucatan, Mexico. A total of 101 isolates representing 53 different taxa were studied. Common fungi such as Fusarium, Pestalotiopsis, Trichoderma and Penicillium were associated with surfaces and were, probably derived from airborne spores. In contrast, unusual fungi such as Rosellinia, Annulohypoxylon, and Xylaria were predominantly identified from mycelium particles of biofilm biomass. Simulating oligotrophic conditions, agar amended with CaCO3 was inoculated with fungi to test for carbonate activity. A substantial proportion of fungi, in particular those isolated from mycelium (59%), were capable of solubilizing calcium by means of organic acid release, notably oxalic acid as evidenced by ion chromatography. Contrary to our hypothesis, nutrient level was not a variable influencing the CaCO3 solubilization ability among isolates. Particularly active fungi (Annulohypoxylon stygium, Penicillium oxalicum and Rosellinia sp.) were selected as models for bioweathering experiments with limestone-containing mesocosms to identify if other mineral phases, in addition to oxalates, were linked to bioweathering processes. Fungal biofilms were seen heavily covering the stone surface, while a biomineralized front was also observed at the stone-biofilm interface, where network of hyphae and mycogenic crystals was observed. X-ray diffraction analysis (XRD) identified calcite as the main phase, along with whewellite and wedellite. In addition, lower levels of citrate were detected by Attenuated Total Reflectance-Fourier-Transform Infrared Spectroscopy (ATR-FTIR). Overall, our results suggest that a diverse fungal community is associated with limestone surfaces insemi-arid climates. A subset of this community is geochemically active, excreting organic acids under quasi-oligotrophic conditions, suggesting that the high metabolic cost of exuding organic acids beneficial under nutrient limitation. Oxalic acid release may deteriorate or stabilize limestone surfaces, depending on microclimatic dynamics.

We studied the relative importance of environmental factors (river flow, nutrients, dissolved organic matter) and organic micro-pollutants [herbicides, insecticides, industrial organic compounds, personal care products, antibiotics and pharmaceuticals, (PhCs)] to the structure and function of epilithic biofilms under various hydrological conditions and land uses., Biofilms from four river basins in the Iberian Peninsula (Llobregat, Ebro, Júcar and Guadalquivir) were analysed in summer-autumn during a wet period and base-flow conditions. The sites experienced different levels of pollution., We used variance partitioning (Redundancy Detrended Analysis, RDA) to link environmental stressors to the structure and function of biofilms, which were assessed as diatom community composition, algal biomass, photosynthetic activity, bacterial density and extracellular phosphatase activity., RDAs showed that industrial organic compounds, herbicides and PhC products were the pollutants most strongly associated with measures of biofilm structure and function, whereas dissolved inorganic nitrogen, dissolved organic carbon and hydrological variability were the environmental factors most strongly associated with biofilm responses., Thicker biofilms developing during base flow were most affected by pollutants and nutrients. In contrast, thinner biofilms developing during periods of high river flow were less affected by chemicals. [ABSTRACT FROM AUTHOR]

Soil and rock surfaces support microbial communities involved in mineral weathering processes. Using selective isolation, fungi were obtained from limestone surfaces of Mayan monuments in the semi-arid climate at Yucatan, Mexico. A total of 101 isolates representing 53 different taxa were studied. Common fungi such as Fusarium, Pestalotiopsis, Trichoderma, and Penicillium were associated with surfaces and were, probably derived from airborne spores. In contrast, unusual fungi such as Rosellinia, Annulohypoxylon, and Xylaria were predominantly identified from mycelium particles of biofilm biomass. Simulating oligotrophic conditions, agar amended with CaCO3 was inoculated with fungi to test for carbonate activity. A substantial proportion of fungi, in particular those isolated from mycelium (59%), were capable of solubilizing calcium by means of organic acid release, notably oxalic acid as evidenced by ion chromatography. Contrary to our hypothesis, nutrient level was not a variable influencing the CaCO3 solubilization ability among isolates. Particularly active fungi (Annulohypoxylon stygium, Penicillium oxalicum, and Rosellinia sp.) were selected as models for bioweathering experiments with limestone-containing mesocosms to identify if other mineral phases, in addition to oxalates, were linked to bioweathering processes. Fungal biofilms were seen heavily covering the stone surface, while a biomineralized front was also observed at the stone-biofilm interface, where network of hyphae and mycogenic crystals was observed. X-ray diffraction analysis (XRD) identified calcite as the main phase, along with whewellite and wedellite. In addition, lower levels of citrate were detected by Attenuated Total Reflectance-Fourier-Transform Infrared Spectroscopy (ATR-FTIR). Overall, our results suggest that a diverse fungal community is associated with limestone surfaces insemi-arid climates. A subset of this community is geochemically active, excreting organic acids under quasi-oligotrophic conditions, suggesting that the high metabolic cost of exuding organic acids beneficial under nutrient limitation. Oxalic acid release may deteriorate or stabilize limestone surfaces, depending on microclimatic dynamics. [ABSTRACT FROM AUTHOR]

Phototrophic river biofilms are microbial assemblages involved in in-stream processes. For a better understanding of N-cycling, the denitrifier community abundance, activity and structure were monitored in natural biofilm assemblages, in two sites exhibiting contrasting nutrient concentrations during a 1-year period. The denitrifier abundance, estimated by quantification of the nirS, nirK and nosZ genes, correlated to biofilm biomass and total bacterial counts. Site-related differences in denitrification activity were observed and the rates were significantly correlated with the nosZ gene copy numbers and biofilm biomass. The denitrifier community structure, assessed by PCR-DGGE of nosZ, differed between sites with only minor differences between sampling occasions, and correlated with the total bacterial community structure. Altogether, these findings suggest that nutrient loading, especially nitrogen, affect both denitrifier community structure and activity. [ABSTRACT FROM AUTHOR]

Abstract: The effect of realistic environmental contamination of diuron on natural epilithic biofilms dwelling bacterial communities and their transformation capacities were investigated by using microcosm experiments. Cobbles carrying biofilms from two sites (“Pau” and “Lacq”) located in areas of contrasting pesticide use (urban and agricultural) on the Gave de Pau river (South-West France) were analysed. The water of the upstream site, Pau, was characterised by fewer pesticides than the water of Lacq, whereas concentrations were higher at Pau. The sampled cobbles were exposed to diuron (10μgL−1) in microcosms. After 3 weeks of exposure, pesticides were analysed and bacterial community structures were assessed with terminal-restriction fragment length polymorphism (T-RFLP). Diuron was biotransformed during contact with biofilms, revealing that these communities contribute to the production of DCPMU (1-(3,4-dichlorophenyl)-3-methylurea) and DCPU metabolites (1-(3,4-dichlorophenyl) urea) in the river ecosystems. Bacterial communities from the most contaminated site appeared to be more resistant to diuron exposure. Correlation analyses combining chemical data with molecular fingerprinting showed that past in situ exposure drove the response of the bacterial communities. [ABSTRACT FROM AUTHOR]

Epilithic bacterial community viability was assessed on natural biofilm assemblages from environmentally contrasting locations over a 17-months period to determine if it reflects environmental conditions or conditions within the biofilm assemblage. Vital state was assessed by membrane integrity using LIVE/DEAD® BacLight™ staining kit. Samples were regularly collected in a large river, up and downstream of a large urban centre. Epilithic biomasses were similar between sites irrespective of the distinct water quality but varied temporarily, peaking up to 48 g AFDM m−2. Bacterial community composition assessed by 16S rDNA based PCR-DGGE significantly differed between sites. Bacterial densities (median of 2.5 × 1011 cell g AFDM−1) were stable whatever the sample origin or biomass. Viable bacterial fractions ranged between 13 and 83% of the total bacterial densities and were correlated with hydrological stability indicators (average of 41.9% during stable water periods, 62.4% during intermediate flow regimes and 50.0% during flow instability) and seasonal parameters. At the river section and epilithic community scales, consistent bacterial densities per unit of biomass could reflect a biofilm assemblage carrying capacity while variable membrane integrity likely integrates changes in the vital state of the community under changing environmental conditions. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Epilithic phototrophic biofilms develop inside Roman Necropolis and Catacombs on rock surfaces exposed to artificial light sources and are composed by a microbial consortium dominated by cyanobacteria. In this work, six non-axenic cultures ofLeptolyngbyasp. strains isolated from biofilms from different Roman hypogea and maintained in cultures from 11 to 20 years were analysed along with their associated bacteria isolated in culture. The employment of PCR-fingerprinting techniques, using HIP1 and ERIC derived primers, allowed the clustering in three groups of the six Leptolyngbya strains and the typing of their isolated bacteria. The bacterial fingerprinting patterns were in agreement with the 16S rRNA gene sequencing and showed the presence inLeptolyngbyaisolates ofPseudomonas, Stenotrophomonas, AgrobacteriumandBacillusrepresentatives that were detected also in biofilms sampled from catacombs. [ABSTRACT FROM AUTHOR]

Elstera litoralis , is a Rhodospirillaceae member which was isolated from the littoral zone of Lake Constance from a stone biofilm using diatom extracellular polymeric substances (EPS) as C source. We present here the draft genome of E. litoralis which has a genome size of 3.83 Mb and 61.2% G + C content. Genome analysis indicated utilization of multiple C substrates explaining its heterotrophic lifestyle as a bacterium present in natural biofilms. Further comparative genome analysis of Elstera with other members of Rhodospirillaceae would be helpful to understand the evolutionary relationships and divergence of hydrobacteria from terrabacteria. [ABSTRACT FROM AUTHOR]

A collection of heterotrophic bacteria consisting of 167 strains was obtained from microbial communities of biofilms formed on solid substrates in the littoral zone of Lake Baikal. Based on the analysis of 16S rRNA gene fragments, the isolates were classified to four phyla: Proteobacteria , Firmicutes , Actinobacteria , and Bacteroidetes . To assess their biotechnological potential, bacteria were screened for the presence of PKS (polyketide synthase) and NRPS (non-ribosomal peptide synthetases) genes. PKS genes were detected in 41 strains (25%) and NRPS genes in 73 (43%) strains by PCR analysis. The occurrence of PKS genes in members of the phylum Firmicutes (the genera Bacillus and Paenibacillus ) was 34% and NRPS genes were found in 78%. In Proteobacteria , PKS and NRPS genes were found in 20% and 32%, and in 22% and 22% of Actinobacteria , respectively. For further analysis of PKS and NRPS genes, six Bacillus and Paenibacillus strains with antagonistic activity were selected and underwent phylogenetic analysis of 16S rRNA genes. The identification of PKS and NRPS genes in the strains investigated was demonstrated among the homologues the genes involved in the biosynthesis of antibiotics (bacillaene, difficidine, erythromycin, bacitracin, tridecaptin, and fusaricidin), biosurfactants (iturin, bacillomycin, plipastatin, fengycin, and surfactin) and antitumor agents (epothilone, calyculin, and briostatin). Bacillus spp. 9A and 2A strains showed the highest diversity of PKS and NRPS genes. Bacillus and Paenibacillus strains isolated from epilithic biofilms in Lake Baikal are potential producers of antimicrobial compounds and may be of practical interest for biotechnological purposes., A collection of heterotrophic bacteria consisting of 167 strains was obtained from microbial communities of biofilms formed on solid substrates in the littoral zone of Lake Baikal. Based on the analysis of 16S rRNA gene fragments, the isolates were classified to four phyla: Proteobacteria , Firmicutes , Actinobacteria , and Bacteroidetes . To assess their biotechnological potential, bacteria were screened for the presence of PKS (polyketide synthase) and NRPS (non-ribosomal peptide synthetases) genes. PKS genes were detected in 41 strains (25%) and NRPS genes in 73 (43%) strains by PCR analysis. The occurrence of PKS genes in members of the phylum Firmicutes (the genera Bacillus and Paenibacillus ) was 34% and NRPS genes were found in 78%. In Proteobacteria , PKS and NRPS genes were found in 20% and 32%, and in 22% and 22% of Actinobacteria , respectively. For further analysis of PKS and NRPS genes, six Bacillus and Paenibacillus strains with antagonistic activity were selected and underwent phylogenetic analysis of 16S rRNA genes. The identification of PKS and NRPS genes in the strains investigated was demonstrated among the homologues the genes involved in the biosynthesis of antibiotics (bacillaene, difficidine, erythromycin, bacitracin, tridecaptin, and fusaricidin), biosurfactants (iturin, bacillomycin, plipastatin, fengycin, and surfactin) and antitumor agents (epothilone, calyculin, and briostatin). Bacillus spp. 9A and 2A strains showed the highest diversity of PKS and NRPS genes. Bacillus and Paenibacillus strains isolated from epilithic biofilms in Lake Baikal are potential producers of antimicrobial compounds and may be of practical interest for biotechnological purposes., (© 2018 Elena Sukhanova et al.)