Emilie Lyautey

Temperature dependence of denitrification in phototrophic river biofilms.

Denitrification is an ecosystem service of nitrogen load regulation along the terrestrial-freshwater-marine continuum. The present study documents the short-term temperature sensitivity of denitrification enzyme activity in phototrophic river biofilms as a typical microbial assemblage of this continuum. Denitrification measurements were performed using the acetylene inhibition method at four incubation temperatures: 1.1, 12.1, 21.2 and 30.9°C. For this range of temperature, N(2)O production could be fitted to an exponential function of incubation temperature, yielding mean (±standard error) activation energy of 1.42 (±0.24) eV and Q(10) of 7.0 (±1.4). This first quantification of denitrification enzyme activity temperature dependence in phototrophic river biofilms compares with previous studies performed in soils and sediments. This demonstrates the high temperature dependence of denitrification as compared to other community-level metabolisms such as respiration or photosynthesis. This result suggests that global warming can unbalance natural community metabolisms in phototrophic river biofilms and affect their biogeochemical budget.

Electroactivity of Phototrophic River Biofilms and Constitutive Cultivable Bacteria

ABSTRACT Electroactivity is a property of microorganisms assembled in biofilms that has been highlighted in a variety of environments. This characteristic was assessed for phototrophic river biofilms at the community scale and at the bacterial population scale. At the community scale, electroactivity was evaluated on stainless steel and copper alloy coupons used both as biofilm colonization supports and as working electrodes. At the population scale, the ability of environmental bacterial strains to catalyze oxygen reduction was assessed by cyclic voltammetry. Our data demonstrate that phototrophic river biofilm development on the electrodes, measured by dry mass and chlorophyll a content, resulted in significant increases of the recorded potentials, with potentials of up to +120 mV/saturated calomel electrode (SCE) on stainless steel electrodes and +60 mV/SCE on copper electrodes. Thirty-two bacterial strains isolated from natural phototrophic river biofilms were tested by cyclic voltammetry. Twenty-five were able to catalyze oxygen reduction, with shifts of potential ranging from 0.06 to 0.23 V, cathodic peak potentials ranging from −0.36 to −0.76 V/SCE, and peak amplitudes ranging from −9.5 to −19.4 μA. These isolates were diversified phylogenetically (Actinobacteria, Firmicutes, Bacteroidetes, and Alpha-, Beta-, and Gammaproteobacteria) and exhibited various phenotypic properties (Gram stain, oxidase, and catalase characteristics). These data suggest that phototrophic river biofilm communities and/or most of their constitutive bacterial populations present the ability to promote electronic exchange with a metallic electrode, supporting the following possibilities: (i) development of electrochemistry-based sensors allowing in situ phototrophic river biofilm detection and (ii) production of microbial fuel cell inocula under oligotrophic conditions.

Rotating disk electrodes to assess river biofilm thickness and elasticity

The present study examined the relevance of an electrochemical method based on a rotating disk electrode (RDE) to assess river biofilm thickness and elasticity. An in situ colonisation experiment in the River Garonne (France) in August 2009 sought to obtain natural river biofilms exhibiting differentiated architecture. A constricted pipe providing two contrasted flow conditions (about 0.1 and 0.45 m s(-1) in inflow and constricted sections respectively) and containing 24 RDE was immersed in the river for 21 days. Biofilm thickness and elasticity were quantified using an electrochemical assay on 7 and 21 days old RDE-grown biofilms (t(7) and t(21), respectively). Biofilm thickness was affected by colonisation length and flow conditions and ranged from 36 ± 15 μm (mean ± standard deviation, n = 6) in the fast flow section at t(7) to 340 ± 140 μm (n = 3) in the slow flow section at t(21). Comparing the electrochemical signal to stereomicroscopic estimates of biofilms thickness indicated that the method consistently allowed (i) to detect early biofilm colonisation in the river and (ii) to measure biofilm thickness of up to a few hundred μm. Biofilm elasticity, i.e. biofilm squeeze by hydrodynamic constraint, was significantly higher in the slow (1300 ± 480 μm rpm(1/2), n = 8) than in the fast flow sections (790 ± 350 μm rpm(1/2), n = 11). Diatom and bacterial density, and biofilm-covered RDE surface analyses (i) confirmed that microbial accrual resulted in biofilm formation on the RDE surface, and (ii) indicated that thickness and elasticity represent useful integrative parameters of biofilm architecture that could be measured on natural river assemblages using the proposed electrochemical method.

Abundance, activity and structure of denitrifier communities in phototrophic river biofilms (River Garonne, France)

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.

Warming-induced changes in denitrifier community structure modulate the ability of phototrophic river biofilms to denitrify

Microbial denitrification is the main nitrogen removing process in freshwater ecosystems. The aim of this study was to show whether and how water warming (+2.5 °C) drives bacterial diversity and structuring and how bacterial diversity affects denitrification enzymatic activity in phototrophic river biofilms (PRB). We used water warming associated to the immediate thermal release of a nuclear power plant cooling circuit to produce natural PRB assemblages on glass slides while testing 2 temperatures (mean temperature of 17 °C versus 19.5 °C). PRB were sampled at 2 sampling times during PRB accretion (6 and 21days) in both temperatures. Bacterial community composition was assessed using ARISA. Denitrifier community abundance and denitrification gene mRNA levels were estimated by q-PCR and qRT-PCR, respectively, of 5 genes encoding catalytic subunits of the denitrification key enzymes. Denitrification enzyme activity (DEA) was measured by the acetylene-block assay at 20 °C. A mean water warming of 2.5 °C was sufficient to produce contrasted total bacterial and denitrifier communities and, therefore, to affect DEA. Indirect temperature effect on DEA may have varied between sampling time, increasing by up to 10 the denitrification rate of 6-day-old PRB and decreasing by up to 5 the denitrification rate of 21-day-old PRB. The present results suggest that indirect effects of warming through changes in bacterial community composition, coupled to the strong direct effect of temperature on DEA already demonstrated in PRB, could modulate dissolved nitrogen removal by denitrification in rivers and streams.